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Biomedical subjects

K Takamura

Publications and source records attributed to K Takamura.

At least 19 recordsLinked to original sources

Nervous network in larvae of the ascidian Ciona intestinalis.

With the use of the monoclonal antibody UA301, which specifically recognizes the nervous system in ascidian larvae, the neuronal connections of the peripheral and central nervous systems in the ascidian Ciona intestinalis were observed. Three types of peripheral nervous system neurons were found: two located in the larval trunk and the other in the larval tail. These neurons were epidermal and their axons extended to the central nervous system and connected with the visceral ganglion directly or indirectly. The most rostral system (rostral trunk epidermal neurons, RTEN) was distributed bilateral-symmetrically. In addition, presumptive papillar neurons in palps were found which might be related to the RTEN. Another neuron group (apical trunk epidermal neurons, ATEN) was located in the apical part of the trunk. The caudal peripheral nervous system (caudal epidermal neurons, CEN) was located at the dorsal and ventral midline of the caudal epidermis. In the larval central nervous system, two major axon bundles were observed: one was of a photoreceptor complex and the other was connected with RTEN. These axon bundles joined in the posterior sensory vesicle, ran posteriorly through the visceral ganglion and branched into two caudal nerves which ran along the lateral walls of the caudal nerve tube. In addition, some immunopositive cells existed in the most proximal part of the caudal nerve tube and may be motoneurons.

Animals

Detection of capillary protein leakage by glucose and indocyanine green dilutions during the early post-burn period.

Overestimation of the plasma volume determined by the indocyanine green (ICG) dilution method (PV-ICG) may occur after burns, since this dye has the potential of extravasation in the presence of the capillary protein leakage. Assuming that the initial distribution volume of glucose (IDVG) consistently indicates the extracellular fluid volume of highly perfused organs including plasma, overestimation of the PV-ICG can be detected by a higher PV-ICG/IDVG ratio. The present study was designed to test whether a higher PV-ICG/IDVG ratio is observed within 24 h post-burn compared to the subsequent days. Ten severely burned adult patients admitted to the ICU were studied through the 2nd post-burn day. The daily IDVG and PV-ICG were calculated using a one compartment model by simultaneous administration of glucose, 5 g, and ICG, 25 mg. Although the IDVG increased on the 1st post-burn day (p < 0.05), the PV-ICG remained unchanged. The PV-ICG/IDVG ratio within 24 h post-burn was significantly higher than that on the 1st post-burn day (p < 0.01). Results indicate that overestimation of the PV-ICG can occur within 24 h post-burn and suggest that simultaneous measurement of the IDVG and the PV-ICG would help predict the generalized capillary protein leakage after burns.

Adult

Measurements of extracellular fluid volume in highly perfused organs and lung water in hypo- and hypervolaemic dogs.

The purpose of this study was to identify whether the central extracellular fluid volume status following hypo- and hypervolaemia can be measured by the initial distribution volume of glucose or by the extravascular lung water. These two estimates were compared with the initial distribution volume of sucrose which has been used as an indicator for the measurement of the extracellular fluid volume. The above three estimates were determined by the administration of glucose, chilled saline and sucrose solutions, before and after haemorrhage (30 mL kg-1), and subsequent fluid load (lactated Ringer's solution 90 mL kg-1). The distribution volumes of glucose and sucrose decreased after haemorrhage and increased after fluid load compared with normovolaemic values, and a linear correlation was obtained between these two distribution volumes (r = 0.93, P < 0.001, n = 36). However, the extravascular lung water remained statistically unchanged throughout the procedure, despite a weak linear correlation with the sucrose distribution volume (r = 0.38, n = 33, P < 0.05). These results indicate that the initial distribution volume of glucose is more useful as an indicator of the central extracellular fluid volume status than the extravascular lung water.

Animals

Time-resolved imaging of protein kinase C activation during sea urchin egg fertilization.

To study protein kinase C (PKC) activation during sea urchin egg fertilization we used three different fluorescent probes specific for PKC, namely, fim-1, which recognizes the catalytic site of the enzyme, and BODIPY- and NBD-phorbol esters interacting with the PKC regulatory domain. We were able to follow PKC activation during the early steps of fertilization, the three different probes giving the same fluorescent pattern. Within 120 s following insemination, the fluorescent signal increased and clustered in the cortical zone of the cell. The process was Ca2+ dependent and was inhibited in the presence of staurosporine, a PKC inhibitor. According to our in vitro probe characterization, this signal increase is due to PKC activation. These findings were further confirmed by Western blot analysis. This initial phase was followed by a rapid decrease which might be attributed to PKC hydrolysis by Ca2(+)-dependent proteases. The kinetics and the site distribution of PKC activation appear in complete agreement with the putative functions previously suggested for PKC during fertilization.

Animals

Determination of p-hydroxymandelic acid enantiomers in urine by high-performance liquid chromatography with electrochemical detection.

High-performance liquid chromatography (HPLC) with electrochemical detection using a chiral ligand-exchange column was developed for the enantioselective determination of p-hydroxymandelic acid (HMA), a metabolite of synephrine, with high sensitivity. A good linear relationship between current ratio and amount was noted for 0.5-500 pmol HMA, with a correlation coefficient of 0.999 for each HMA enantiomer. The relative standard deviation (R.S.D.) was 1.6% at 100 pmol d-HMA and 2.2% at 100 pmol l-HMA. The detection limit of each HMA enantiomer was 0.5 pmol (signal to noise ratio, S/N = 3). By this method, HMA in Citrus unshiu and in urine following the ingestion of C. unshiu was determined. Although no HMA was found in c. unshiu, d- and l-HMA were present in urine after the ingestion of C. unshiu. The time courses of HMA and conjugated synephrine enantiomers excreted in urine following the ingestion of C. unshiu for 24 h could be monitored. This method should prove applicable to the study of synephrine metabolism.

Adult

Determination of acid values of fats and oils by flow injection analysis with electrochemical detection.

A new method using a flow injection system with electrochemical detection was developed to determine acid values of fats and oils. VK3 (2-methyl-1,4-naphthoquinone) solution, i.e., ethanol containing 3 mM VK3 and 38 mM LiClO4, was used as the carrier solution. Flow signals were monitored at -0.33 V vs. Ag/AgCl. For preparation of a sample solution, an oil sample was completely dissolved in VK3 solution, or fatty acids were extracted from the sample into this solution. Aliquots (5 microliters) of the sample solution were injected into the flow injection system. Acid values were determined based on flow signals for 14 samples and the results were found to be consistent with those by potentiometric titration. Relative standard deviation was less than 2%. Samples were processed at the rate of 60 h-1. The stability of fish and cod liver oils was followed by measuring acid values for 8 weeks. This method proved to be a simple and rapid means for acid value determination.

Acids

Peripheral primitive neuroectodermal tumor involving the paravertebral and retroperitoneal regions.

A rare case of peripheral primitive neuroectodermal tumor (PNET) is reported. A 68-year-old woman complaining of lumbago was admitted to our hospital. Diagnosis was made based on pathological findings characterized by Homer Wright-type rosettes. Ultrastructural examination showed the presence of neurosecretory granules and short cytoplasmic processes, which were highly suggestive of neural differentiation. Chromosomal analysis of the neoplastic cells revealed translocation (11;22)(q24;q12), which is often found in Ewing's sarcoma and Askin tumor. These results strengthen the hypothesis of a common histogenesis for these small round cell tumors, and suggest common oncogenesis for these neoplasms.

Aged

Intravascular lymphomatosis diagnosed by transbronchial lung biopsy.

Intravascular lymphomatosis is a rare lymphoma presenting a variety of symptoms due to proliferation of tumour cells within blood vessels in the brain, the skin and other organs. This disease is generally considered to be highly malignant, but to be relatively susceptible to combined chemotherapy, when diagnosed in the early stage. We describe a case of intravascular lymphomatosis, presenting with diffuse interstitial shadows on chest radiographic image, which could be diagnosed by transbronchial lung biopsy. The patient showed a good response to combined chemotherapy. We propose that transbronchial lung biopsy is a useful procedure for the diagnosis of intravascular lymphomatosis.

Antineoplastic Combined Chemotherapy Protocols

[Usefulness of exercise electro-cardiography in a THP (Total Health Promotion Plan)].

We conducted ergometer exercise electrocardiography (ergometry) on 3,477 subjects in a THP (Total Health Promotion Plan). One hundred cases in which abnormal findings were detected by ergometry were analyzed. In the hundred cases there were 3 patterns: abnormal ST change, 50 cases; abnormal reaction, 22 cases; and extreme increase in blood pressure, 28 cases. Electrocardiograms (ECG) in 78 of these 100 cases indicated no abnormalities. Of the 31 subjects who underwent further examinations, in 18 cases abnormal findings were detected, and further observation or treatment was necessary. They were over two thirds of the 26 cases requiring observation or treatment on further examination. In other words, exercise electrocardiography revealed more than 3 times as many problem cases as electrocardiography only. One hundred and four cases were analyzed, and among them abnormal findings on ECG made further examination or treatment necessary. Of 68 subjects with an abnormal ECG and who needed to undergo exercise electrocardiography, 51 (75%) had no need to undergo further examinations, because there were no abnormal findings on ergometry in the THP. Of the 104 subjects who underwent ECG examination at rest, 51 no longer needed to waste time, effort and expense on further medical evaluation. Ergometry in a THP serves as a medical check and as a means to decide the strength of exercise before the initiation of exercise training, which is very important in preventing coronary artery disease, rather than in detecting the disease. Ergometry is expensive and it takes a lot of time and labor, but it is necessary in ensuring the safety of exercise training and in prescribing proper exercise. This analysis has shown that ergometry in THP is very useful and cost effective in improving the accuracy of health examinations.

Adult

Selective determination of chloride and bromide ions in serum by cyclic voltammetry.

Voltammetric determination of chloride and bromide ions in serum was made using a one-body type Ag electrode, whose surface was covered with a dialysis membrane for preventing interference from proteins. This determination was based on measurement of the charge of the reduction wave of silver halide formed on the Ag electrode surface in a halide ion solution during a cathodic potential sweep. In this method, depending on the potential sweep range, the charge of the reduction wave of AgI, AgI + AgBr, or AgI + AgBr + AgCl can be measured. Here, the method was used to determine the chloride and bromide ions in artificial serum. Linear concentration ranges were 1.0 x 10(-2)-2.4 x 10(-1) M for chloride ions and 5.0 x 10(-3)-1.8 x 10(-2) M for bromide ions. The correlation coefficient was 0.999 in each case. The relative standard deviation for chloride ion (1.0 x 10(-1) M) was 1.9% (n = 10), and that for bromide ion (1.5 x 10(-2) M) was 2.5% (n = 10). Bromide ions at a relatively high concentration as the component of serum had no effect on the determination of the chloride ion in the serum. This method is thus useful for the selective determination of chloride and bromide ions in serum.

Blood Substitutes

Determination of synephrine enantiomers in food and conjugated synephrine in urine by high-performance liquid chromatography with electrochemical detection.

Determination of synephrine enantiomers was made with HPLC with electrochemical detection using a chiral ligand-exchange column (Sumichiral OA-6000). The calibration curve for each enantiomer showed good linearity (r = 0.999) at 1.0-500 pmol injected with a detection limit of 1.0 pmol (signal-to-noise ratio (S/N) = 3). Relative standard deviation (RSD) was 1.8% at 50 pmol d-synephrine and 1.6% at 50 pmol l-synephrine. The contents of synephrine enantiomer in food such as Citrus unshiu fruit, orange juice, and marmalade were determined. The present method was also used to determine conjugated synephrine enantiomers in urine following the ingestion of C. unshiu pulp. l-Synephrine in fruit was converted to the conjugated form of synephrine, and l-synephrine underwent chiral inversion to d-synephrine in vivo.

Adult

Sensitive densitometry for the determination of platelet-activating factor and other phospholipids in human tears.

A sensitive TLC method is reported for the determination of the platelet-activating factor (PAF) and other phospholipids in human tears. Tear samples absorbed on filter-paper were subjected to diatomite column extraction with chloroform-methanol. The eluent containing phospholipids was spotted on a silica gel plate. After removing lipids other than phospholipids by pre-development with hexane-diethyl ether (4 + 1 v/v), individual phospholipid separation was carried out by development with chloroform-methanol-water (65 + 35 + 7 v/v). Phospholipase C and then alkaline phosphatase solutions were sprayed on the TLC plate at 45 degrees C to liberate phosphate from each phospholipid. By spray application of a mixture of ammonium molybdate and Malachite Green, the liberated phosphates appeared as blue-green spots of molybdophosphate-Malachite Green aggregate on a yellow-brown background. The absorbance of each spot on the plate was measured at 620 nm with a densitometer. The peak area was found to be linearly related to PAF content in the range 2-100 pmol per spot, the RSD being 2% (n = 7). Approximately 86% of standard PAF added to tears was recovered. By this method, lysophosphatidylcholine (lysoPC), phosphatidylcholine (PC) and phosphatidylethanolamine (PE) in human tears could also be detected with high sensitivity. Each phospholipid in healthy human tears showed a nearly constant content; PAF, lysoPC, PC and PE were present at levels of 26.2, 42.3, 10.0 and 19.7%, respectively.

Chromatography, Thin Layer

Motor and sensory disability has a strong relationship to induction dose of thiopental in patients with the hypertropic variety of Charcot-Marie-Tooth syndrome.

In a prospective study, we determined the anesthetic induction dose of thiopental and the clinical variables influencing the appropriate induction dose of thiopental in 20 patients with the hypertrophic variety of Charcot-Marie-Tooth syndrome (CMT). As controls we chose 50 patients without CMT. Motor disturbance was evaluated in terms of muscle weakness of the distal lower and upper extremities. We examined sensory disturbance by evaluating loss of sensation in the index finger and great toe. The preinduction cardiac output was measured by echocardiography. Anesthesia was induced with repeated injections of 50 mg thiopental. The minimum induction dose of thiopental (MID) was confirmed when the eyelash reflex ceased. We maintained anesthesia with enflurane and nitrous oxide. The 95% confidence interval of the MID in patients used as the controls was 2.5-4.9 mg/kg. The MID in 11 patients with CMT was less than 2.5 mg/kg. MIDs in the patients with CMT were significantly smaller than those of the control patients (P < 0.0001). Also we found a strong relationship between the MID and the severity of both motor and sensory disturbances (P = 0.003 and 0.002, respectively). There was no relationship between the MID and other clinical variables, such as age, gender, inherited type, body weight, and preinduction cardiac output. Because delay in the recovery from anesthesia can be caused by an inappropriate dose of thiopental in CMT patients in whom motor and sensory function is seriously impaired, the dose of thiopental probably should be reduced and based on the individual patient's response.

Adolescent

[Toxic shock-like syndrome with Streptococcus pyogenes in a pleural effusion].

A 58-year-old woman was admitted to the hospital because of fever and hypotension. That night, shock developed. On the second hospital day, a chest roentgenogram showed retention of pleural fluid and the group A-beta hemolytic organism Streptococcus pyogenes was detected in the effusion. The toxic shock-like syndrome was diagnosed. The patient recovered with artificial ventilation, administration of antibiotics, and blood purification. In this patient, the type of pyrogenic exotoxin was B + C.

Female

Determination of barnidipine in human serum and dog plasma by HPLC with electrochemical detection.

Barnidipine is a 1,4-dihydropyridine calcium antagonist. HPLC was conducted on a polybutadiene coated alumina column using an alkaline mobile phase and an electrochemical detector to determine the content of this drug in serum and plasma. A good linear relationship between barnidipine concentration and peak height was found in 5-500 ng/ml with a correlation coefficient of 0.998. The detection limit was 1 ng/ml. The within-day and day-to-day variations were examined for control human serum. Relative standard deviation of within-day assay for serum spiked with 10 ng/ml barnidipine.HCl was 6.9% and the recovery was 104%. A pharmacokinetic study was made in which the time course of barnidipine in dog plasma was followed.

Animals

Evaluation of carcinogenicity and chronic toxicity associated with orthopedic implants in mice.

The carcinogenicity and chronic toxicity of 316L stainless steel, nickel, Ti-6A1-4V, hydroxyapatite (HA)-coated Ti-6A1-4V, aluminum oxide containing yttrium oxide, and zirconium oxide containing yttrium oxide were evaluated by implanting solid rods of each material in the thigh muscle of C57BL/6N mice for 24 months. Nickel alloy showed high carcinogenic and toxic potencies, whereas other materials showed no evidence of them. Tumors retaining nickel alloys were malignant fibrous histiocytoma or fibrosarcoma. In some cases, lymphomata that seemed to develop spontaneously were found around the implants because lymphocytes were known to accumulate in chronic inflammatory lesions, and this phenomenon also might be applied to lymphoma.

Alloys

[Spectrophotometric determination of uric acid in serum using a titanium (IV)-porphyrin complex].

Aqueous solution of oxo[5,10,15,20-tetra(4-pyridyl)porphyrinato]titanium (IV) complex, a Ti-TPyP reagent, was found to be very useful for the spectrophotometric determination of hydrogen peroxide. The reagent (lambda max 432 nm) reacts with hydrogen peroxide to form a monoperoxocomplex, resulting in a significant decrease of the absorbance at 432 nm. The decrease (delta A) in absorbance was proportional to the concentration of hydrogen peroxide. The Ti-TPyP reagent was successfully applied to the assay of uric acid in the serum, using uricase to produce hydrogen peroxide through enzymatic oxidation. Using only 5 microliters serum, a linear relationship was obtained between delta A and uric acid concentration in the serum ranging from 5 x 10(-6) to 1 x 10(-3) M. The apparent molar delta A of uric acid was 2.2 x 10(5) M-1 cm-1. The relative standard deviation of repeated runs (n = 8) was 2.8% at 3.77 x 10(-4) M uric acid. The analytical recovery of uric acid (5 x 10(-4) M) added to the serum was 96.8 to 105.0%. No pre-concentration and deproteinization were required to determine uric acid in the serum by the present method because of the high sensitivity and selectivity of the Ti-TPyP reagent for hydrogen peroxide.

Gout