[An assessment of nosocomial infections of methicillin-resistant Staphylococcus aureus based on coagulase typing and phage typing].

An epidemiologic investigation was carried out in Ogaki Municipal Hospital to clarify the status of nosocomial MRSA Infection between 1989 and 1991. In 1989, coagulase type IV, enterotoxin A-producing, and phage group I strains, which were highly resistant to multiple antibiotics and isolated in the internal wards, accounted for 43.4% of all MRSA strains clinically isolated in the entire hospital. In 1990, coagulase type II strains that were sensitive to GM but resistant to FMOX and IPM increased. There were significant differences in the frequency of detection of various strains among wards, suggesting an inter-ward variation in MRSA strains. Changes in environmental strains reflected those in clinical strains. The findings suggest the necessity of measures not only for long-hospitalized MRSA carriers themselves but also for the environment of patients, medical staff, and those taking care of patients.


[Genomic DNA fingerprinting by pulsed-field gel electrophoresis for study of nosocomial infection caused by methicillin-resistant Staphylococcus aureus].

In this study, we have compared genomic DNA fingerprintings among isolates of methicillin-resistant Staphylococcus aureus (MRSA) by using pulsed-field gel electrophoresis (PFGE). Chromosomal fragments digested with Sma I were most suitable for the PFGE separation, giving 15-20 fragments of sizes ranging from about 30-1,500 kb. Thirty-one distinctive fragment patterns were identified in 111 isolates of MRSA. On the basis of the genomic typing by PFGE, we performed an epidemiological investigation of an outbreak of nosocomial MRSA infection among 18 inpatients in Nagoya University Hospital. Ten types of chromosomal digestion were identified in 21 isolates of MRSA. The clarity and polymorphism of the chromosomal digestion patterns enabled us to discriminate between isolates. Classification of the genomic DNA fingerprinting patterns by PFGE is therefore proposed as a useful method for investigating the source, transmission, and spread of nosocomial MRSA infections.


[Carrier state and comparative study of methicillin-resistant Staphylococcus aureus from the staff and patients].

The nasal carriage of Staphylococcus aureus in 529 staff was screened. S. aureus from staff and patients whose minimum inhibitory concentration (MIC) of methicillin was larger than 12.5 micrograms/ml by agar dilution was defined as Methicillin-resistant S. aureus (MRSA). Coagulase typing was performed by DENKA SEIKEN kit. Hands of ward staff were screened before and after contact with the MRSA carriers and after hand washing. The nasal acquisition rate of S. aureus and MRSA for staff was 27.6% and 8.5%. The rate of ward staff for nasal carriers of MRSA was 91.1%. Coagulase type II strains from ward staff and inpatients were dominant. In some cases MRSA was detectable after hand washing: In MRSA infection ward staff played a dangerous role as a vector.


[Nucleic acid probes in infectious diseases].

DNA probes are then newest diagnostic reagents now in clinical use to detect or specify infectious microorganisms. The fundamental aspects of DNA probes and their clinical applications are reviewed to provide the clinician new information on the recent progress in infectious diseases.


[Biosensors and clinical analysis].

Method for the selective determination of organic compounds in biological fluids, such as blood, are very important in clinical analyses. Most analyses of organic compounds can be performed by spectrophotometric methods based on specific enzyme-catalyzed reactions. However, on account of the complicated procedure, these methods cannot be applied directly to bed-side monitoring. Alternatively, biosensors based on enzymes and electrochemical transducers possess excellent sensitivity for biological substrates and can determine directly a single compound in a biological fluid without need for a prior separation step. Therefore, in several years, many kinds of biosensors have been developed, and implantable microbiosensors for a body organ have also been reported. In the near future, an artificial pancreas fabricated by micromachine technology will be developed.


[Application for medical field of enzyme sensor].

Various portable glucose sensors have been developed and used for self-measurement of blood glucose levels for diabetes. Enzyme sensors will become one of a useful technology for assay of various substances and be applied to analysis for clinical chemistry.


[Utilization of an enzyme-sensor in clinical medicine].

The utilization and merit of an enzyme-sensor for measuring serum- or urine-components were reported. The enzyme sensor (enzyme-electrode) is an H2O2-electrode, O2-electrode or ion selective electrode connected with an enzyme-immobilized nylon tube. First, the principle of measurements with an enzyme sensor was reported. Secondly, and H2O2 electrode which had been planned and developed in our laboratory was introduced. The outline of our apparatus and the procedure of enzyme-immobilization onto a nylon tube wall were explained. Then the results obtained from quantitative analysis of serum- and urine-components were shown. For the analysis by our apparatus, 3-60 microliters of serum or urine was used and incubation time was approximately 5 min. The results obtained by our apparatus coincided with those obtained by other methods (r = 0.981-0.997). Thirdly, the usefulness of commercial instruments with an enzyme-sensor for determining glucose- or lactic acid-content was described. The results (glucose and lactic acid-contents) measured by desktop-size instruments showed a high lineality and good reproducibility (CV = 0.6-1.5% in glucose and 0-0.7% in lactic acid). A good correlation was observed among each instrument (r = 0.999). The results by card-size instruments, GLUTEST E or ExacTech, showed a lower reproducibility (CV = 4.1-6.5%) and correlation (r = 0.95-0.98) than those by desktop-size instruments.


[Biosensing techniques for the laboratory medicine].

Biosensing techniques are utilized in micro analysis of the biological constituents with an integrated device. Their applications to laboratory medicine are of great interest. Recently, several types of micro electrodes or ion sensitive field effect transistors (ISOFET) were developed. For fabrication of the biosensor selection of the proper bio-element and transducer to assay our sample, is important. In this article we will indicate some items on the future prospect of biosensing techniques for laboratory medicine.


[Clinical usefulness of essential laboratory tests].

Clinical usefulness of Essential Laboratory Tests (1), proposed by Japan Society of Clinical Pathology, was retrospectively evaluated in 500 outpatients who first visited the medical clinic of a prefectural general hospital. Sixty percent of the patients were diagnosed with gastrointestinal disease or respiratory disease. Performance rates of urinalysis, blood counts, serum total protein, A/G ratio, C-reactive protein, erythrocyte sedimentation rate and fecal occult blood were 87.6%, 55.8%, 49.6%, 48.8%, 37.8%, 10.0% and 5.6%, respectively. Abnormal test results rate of urinalysis, blood counts, serum total protein, A/G ratio, C-reactive protein, erythrocyte sedimentation rate and fecal occult blood were 12.3%, 12.9%, 14.1%, 5.7%, 21.6%, 22.6% and 25.9%, respectively. The clinical diagnoses and the abnormal test results indicated that the patients general condition was assessed properly from the A/G ratio, 92% of patients with proteinuria were transient cases and urobilinogen has poor sensitivity and specificity for the diagnosis of liver diseases.


["Essential laboratory tests" in primary care medicine--experiences in Comprehensive Medicine, National Defense Medical College].

The "essential laboratory tests" were applied to 1,026 new patients visiting the outpatient unit of Comprehensive Medicine, National Defense Medical College, to determine the usefulness of these tests in primary care medicine. The "essential laboratory tests" have contributed to the establishment of the initial diagnosis or confirmation of the "tentative initial diagnosis", and to the estimation of the nature or degree of seriousness of the disease. In addition, other diseases not related to the patient's chief complaint could be screened in 32% of the new patients examined with these tests. The "essential laboratory tests" were also found to be useful for the estimation of the presence of malignant tumors in 12 patients with various kinds of malignancies as well as the estimation or evaluation for such conditions as infection or inflammation, anemia, liver or renal dysfunction, and the ambulatory screening of metabolic diseases including hyperlipidemia, liver or urinary tract diseases. In conclusion, simultaneous performance of the "essential laboratory tests" with the history taking and the physical examination will provide important medical information in primary care medicine.


[The effectiveness of the essential laboratory tests in daily primary care--view of a physician at a private clinic].

I put into practice, "The essential laboratory tests (a proposal)" published by Japan Society of Clinical Pathology in 1989 on new patients who visited my clinic. It was useful not only for screening of diseases, but enhanced the probability of diagnosis. I could discover hidden abnormalities unrelated to the patient's complaints. Therefore, the condition of the patient could be judged theoretically and scientifically by the use of the essential laboratory tests.


[Proper use of laboratory tests from a perspective of health insurance administration].

The national medical expenditure increases at a rate of 1 trillion yen a year, and the expenditure for 1992 is estimated to be about 23 trillion yen. One of the most important jobs of the Health Insurance Administration is to control the medical expenditure. Every time the medical fee is amended, laboratory tests are always the target of the cost containment. In the past, the biochemical tests have been separated into the cost for performance of test, and the cost for assessment of the results, the cost of tests for specific diseases has been comprised, or fixed, and the fee of EKG, EEG, Echo etc after the first has been forced to be reduced. These are all administrative policies, but ultimately, they must be selected properly by the physician who requests these tests. In the future, a system should be established which enables the physician to efficiently (cost-efficiently) select the most appropriate test. For this purpose, undergraduates and post-graduates should learn the concepts of clinical epidemiology and clinical economics. These academic disciplines should be incorporated to increase the value of the treatment. Furthermore, the physician should be given economical incentive of proper use of medical resources.


[Current status and problems of platelet function tests].

Platelet function tests have been utilized for a long time as a useful tool for the diagnosis of qualitative platelet disorders. However, recently reports suggest that platelet function tests currently available in routine laboratory do not necessarily reflect in vivo hemostatic states. First of all, bleeding time could sometimes be non-informative for the following reasons; 1) Duke method, the most popular method in Japan, has poor reproducibility, 2) there is no appropriate method for monitoring of aspirin ingestion, 3) prolongation of bleeding time does not correlate with the amount of blood loss during surgery. Platelet adhesion is still measured by the ability of platelets to be retained on glass beads. However, this test is unable to detect selectively platelet adhesion. Thus, the test in which platelets can exclusively adhere to subendothelial components such as collagen, may be required. Platelet aggregation has been mostly analyzed by the changes in light transmittance in platelet rich plasma (PRP) with a platelet aggregometer. However, this test totally depends on the platelet count in PRP or duration of time after taking blood from patients. Moreover, platelet aggregation in this system is optimally observed and is hardly detectable when the in vivo Ca++ concentration has been chelated. Furthermore, the test could not detect small platelet aggregates, thus being unable to measure the early phase of platelet aggregation. These observations suggest that more simple and specific tests should be developed and become available in routine laboratory.


[Platelet adhesion and adhesive proteins--how to measure the platelet adhesiveness].

We examined, platelet adhesive function in vivo by comparing the platelet retention rate (PRR) with other laboratory tests. In 71 patients, PRR did not correlate with bleeding time (BT), ristocetin-induced platelet agglutination (RIPA), vWF:AG, vWF:RCo, FVIII:C, fibrinogen, platelets counts or platelet aggregation tests. Among them, the coincidence rate of PRR with BT seems relatively better than that of other tests. In patients with Bernard-Soulier syndrome (BSS), PRR, RIPA and high shear-induced platelet aggregation (SIPA), measured using a cone-plate type instrument, were decreased, while platelet adhesion in a static system and SIPA at a low shear rate were normal. In patients with Glanzmann's thrombasthenia (TA), PRR and SIPA were decreased at both high and low shear rates, while RIPA and platelet adhesion were normal in a static system. In patients with deficient glycoprotein Ia/IIa complex which is one of the putative receptors for collagen, platelet adhesion in a static system is markedly decreased. In patients with von Willebrand disease, the pattern of platelet adhesion tests was similar to that of BSS patients, while in patients with afibrinogenemia, it was similar to that of TA patients except for SIPA at a high shear rate. The various experiments using monoclonal antibodies support these abnormalities. These findings, suggest that PRR is a suitable screening test, but further rapid and easy tests will be necessary to diagnose the defects of platelet adhesion at various shear rates.


[Clinical evaluation of four tumor markers (CEA, TPA, CA50 and CA72-4) in colorectal cancer].

In a total of 194 cases, consisting of 86 cases of colorectal cancer undergone operation later, 7 cases of non resectable cancer, 34 cases of recurrence, 43 cases of NED (no evidence of a recurrence after radical surgery for colorectal cancer) and 24 cases of benign colorectal disease, serum CEA, TPA, CA50 and CA72-4 levels were determined. The positivity rate was high for all four markers in stage V cases among 86 cases of colorectal cancer, and in cases of non resectable cancer and cases of recurrence. The highest positive rate was obtained with CEA. On the contrary, in cases of stage I to IV the positivity rates of these four tumor markers were as low as 0 to 34.8%. Out of 127 cases of colorectal cancer excluded of 43 NED cases, 52 cases were negative for all four tumor markers and 14 cases were positive only for CEA. In 49 cases, CEA and at least one of the other three tumor markers gave positive results. In 12 cases, CEA gave negative results and at least one of the other three tumor markers positive results. In conclusion, measurement of blood levels of these tumor markers is limited of its usefulness in early diagnosis of colorectal cancer. However, in the diagnosis of advanced stage and of recurrence during the postoperative follow-up period the measurement of tumor markers provides useful information. CEA is most sensitive among the four tumor markers tested and any combination of these four markers is not advantageous because of an increase in false positivity rate.


[Increased serum levels of human cardiac myosin light chain 1 in patients with renal failure].

In this study, we measured cMLC1 concentration in serum and urine from patients with acute myocardial infarction (AMI), chronic renal failure (CRF), and various grades of renal dysfunction (RD) in comparison with normal controls, by using enzyme immunoassay (EIA) with monoclonal antibody, and attempted to elucidate the mechanism of increased serum level of cMLC1 in patients with renal failure. The serum level of cMLC1 of CRF patients under maintenance hemodialysis (HD) was 20.3 +/- 19.6 ng/ml, markedly higher than normal controls (0.54 +/- 0.55 ng/ml). The patients with RD and CRF under conservative therapy had higher serum cMLC1 level than normal controls especially in advanced CRF, while each value not correlating with their creatine clearance (Ccr). cMLC1 in urine was detectable in only two cases with AMI accompanied with CRF or RD. In addition, immunohistological studies of renal biopsy specimens from RD patients did not show cMLC1 deposits in glomerulus. These results suggest that cMLC1 is assumably filtered through the glomerulus, and then absorbed in the renal tubule.


[Identification of two different genetic mutation associated with silent phenotypes for human serum cholinesterase in Japanese].

Two different gene mutations associated with the silent phenotype for human serum cholinesterase were demonstrated. DNA from five individuals with silent gene phenotype of three unrelated Japanese families was amplified by the polymerase chain reaction (PCR) and analyzed by direct sequencing. The first instance demonstrated a G----C transversion at codon 365 from GGA (Gly) to CGA (Arg), which was seen in three individuals of the two families. This mutation was resulted to create a new Taq 1 restriction site (TCGA). The second mutation was shown by a double heterozygous condition with two different silent gene mutations in two members of remaining one family. These mutations were as follows: 1) one type was a frameshift mutation, in which an extra A was inserted in codon 315 (ACC----AACC) to create a new stop codon at position 322 and 2) the other was the same point mutation at codon 365 as seen in the first instance. These results indicated that many silent variants can be distinguished by direct sequence analyses of genomic DNA.


[Detection of methicillin-resistant Staphylococcus aureus using PCR and non-radioactive DNA probe].

Two commonly used diagnostic tests for Staphylococcus aureus (MRSA) are cultivation of bacteria from clinical samples on mannitol-salt agar plate or on MRSA-screen agar plate with oxacillin. However, the use of PCR and DNA probes is considered more rapid and sensitive, as staphylococcal cells have high resistance to beta-lactam due to the novel penicillin-binding protein, PBP2'. Therefore, three different pairs of DNA primers (P1F-P1R, P2F-P2R and P3F-P3R) complementary with unique regions of the MRSA PBP2' gene were synthesized for use in polymerase chain reactions with DNA of MRSA. Amplified target DNA of 466, 480 and 630 bp could be resolved on ethidium bromide-stained gels, and hybridized with DNA probes conjugated to alkaline phosphatase. When applied to pure cultures on the MRSA screen agar, the DNA probes detected MRSA in 180 of 196 (P1F-P1R), 72 of 83 (P2F-P2R) and 66 of 71 (P3F-P3R) culture-positive specimens (accuracy range, 86.7-93.0%), while 61.6-89.3% of MRSA were detectable in culture-positive specimens streaked on the mannitol-salt agar. When applied directly to clinical samples, this DNA probe assay detected MRSA in culture-positive specimens within an accuracy range of 50.0-79.3%. It was thus clear that comparison of cultivation and DNA hybridization results yields good correlation with respect to detection of MRSA. These results suggest that the DNA probe assay may be useful in complementing existing culture techniques.


[Clinicobacteriological study of Pasteurella multocida infection as a zoonosis. (2)--Comparisons of P. multocida separated from patients and their pet dogs and cats].

A total of 17 strains of Pasteurella multocida, of which 13 were isolated from patients treated at Nihon University Itabashi Hospital or Nihon University Surugadai Hospital between April, 1984 and March, 1991 and 4 from 1 dog and 3 cats kept by the patients, were evaluated with respect to their biochemical properties, sensitivity to drugs, and serotype. The isolated strains were all considered to be Pasteurella multocida subsp. multocida because of the agreement of their responses to indole, sorbitol and dulcitol with those of this subspecies, except for 1 sorbitol-negative strain of Pasteurella multocida subsp. septica isolated from 1 patient who had been bitten by a cat. All the isolated strains showed high sensitivities to various drugs. The serotype was capsular type A, which is often observed in cats and dogs, in 7 strains, which consisted of 6 of the 7 strains derived from the airway of the patients and 1 of the 6 strains derived from bit or scratch wound. The remaining strains could not be classified. Five morphological types, namely 1, 3, 3.8, 6, and 8 were observed. In 2 patients, Pasteurella multocida subsp. multocida of the same serotype was also isolated from their cats. One of these patients had intimate contact with the cat including kissing. Our findings suggest that: 1) Pasteurella multocida subsp. multocida has been responsible for most conventional cases of Pasteurella multocida infection. 2) Strains isolated from patients differ in the capsular type according to the disease.(ABSTRACT TRUNCATED AT 250 WORDS)


[Simple method for measurement of erythrocyte osmotic fragility].

We developed a simple and quantitative method for determination of erythrocyte osmotic fragility, which is based on a logarithmic linearization of osmotic fragility curve and a calculation of three parameters, that is, C50 (the osmolarity causing 50% hemolysis), beta (the slope of the curve) and H (% hemolysis before treatment). By making use of personal computer program, calculation of the parameters and print-out of reports are automated. As the results, it takes only 30 min to finish all procedure. This method is very useful particularly for the laboratory screening of hereditary spherocytosis.


[Correction of haptoglobin contents in hemolytic specimen with addition of saturated hemoglobin].

Measurement of serum haptoglobin by immunoturbidimetry was markedly interfered with hemolysis because of haptoglobin-hemoglobin complex formation. Turbidity formed with haptoglobin and immunoglobulin complexes was gradually decreased by the addition of hemoglobin in the reaction mixture, and reached to a certain fixed levels when serum haptoglobin were saturated with hemoglobin. The turbidity derived from saturated haptoglobin and hemoglobin complex closely correlated with the haptoglobin level without hemoglobin. Thus, we could measure the corrected haptoglobin level in hemolytic specimen by the saturation of hemoglobin in reaction mixture.


[Key note comment on the determination of glomerular filtration rate].

Glomerular filtration rate (GRF) is the most fundamental renal function. Determination of GFR is the most important examination in clinical nephrology. Cin is accepted as the gold standard of GFR, but, continuous infusion and the determination method of inulin (method of anthrone) are very troublesome. Interference of glucose in the serum and urine of patients with diabetes mellitus on the determination of inulin should be avoided. Ccr is widely used in the clinical field. Recently, the dynamics of creatinine in the body have been clarified. Variation of pool, generation rate, extrarenal excretion and tubular excretion of creatinine are found to change daily in the patient. Additionally, classical Jaffé's reaction involves non creatinine chromogen particularly in the serum. Though Jaffé's reaction with kinetic analysis improves the reliability of this reaction, the result is still unsatisfactory. There are several methods of enzyme assay are now using an autoanalyser, but, if we use many enzymes and reagents, the reliability of this method could not exceed that of Jaffé's method. Recently, HPLC is recommended as the standard method. 24-hr urine collection is also troublesome in the measurement of Ccr. I have found a good correlation between the estimated Ccr using Cockcroft and Gault's formula and Cin. Such prediction using serum creatinine concentration and few parameters should be developed. Radiolabelled agents have been widely used for the determination of GFR. However, in Japan, the methods of clearance of non radiolabelled contrast agents should be developed. Furthermore, a physical determination of GFR is necessary. In this workshop, these issues will be discussed by each speaker.


[Measurement of renal plasma flow and glomerular filtration rates in diabetic subjects].

To clarify the problem in the measurement of renal plasma flow and glomerular filtration rates in diabetes, the effect of glucose on the determination of para-aminohippuric acid (PAH) and inulin was examined. The concentration of urinary PAH in glucosuric diabetic subjects decreased after the storage of urine samples because of the glycation of PAH. Therefore, glucose must be removed by the acid treatment before the determination of the concentrations of urinary PAH. Since glucose can interfere with the assay of inulin, the sample must be treated with NaOH prior to the determination of the inulin concentration. GFR of the subjects with type 2 diabetes was next examined. GFR in the subjects with a duration of diabetes less than 10 years was significantly higher than that in the subjects with a duration of diabetes more than 10 years. Thus, the subjects with short-term type 2 diabetes may present with hyperfiltration similar to the subjects with short-term type 1 diabetes.


[A method for the simultaneous determination of iothalamic acid and creatinine clearance by coupled-column liquid chromatography].

[125I]iothalamic acid clearance is used for the estimation of glomerular filtration rate. Therefore, we developed a coupled-column liquid chromatography for the simultaneous determination of iothalamic acid and creatinine in human serum and urine samples to calculate their clearance. The liquid chromatograph consisted of an automatic injector, four columns and four switching valves. Timing of sample injection and switching of valves were controlled by a combination of timer units. Diluted samples were injected into a column of weakly acidic ion exchanger (Asahipak ES-502C, 30 degrees C, C1), and iothalamic acid excluded from the column was purified by gel chromatography on a hydrophilic gel (Asahipak GS-320H, 50 degrees C, C2) and then by ion exchange chromatography on a weakly basic anion exchanger (Asahipak ES-502N, 50 degrees C, C3). Creatinine that was eluted from C1 after iothalamic acid was transferred to Asahipak GS-320H (37 degrees C, C4) and C1 was backflushed until 10 min before the next sample was injected. Samples were injected every 55 min. The mobile phase was a sodium propionate buffer prepared by dissolving 0.035 mol of NaOH and 0.35 mol of propionic acid in 1 kg of ultra pure water, and its flow rate was 1.1 ml per min. Creatinine clearance calculated from the data obtained by this method was higher than that obtained by Jaffé's method, because serum creatinine concentrations obtained by this method were lower than those obtained by Jaffé's method. Iothalamic acid clearance (y) calculated from the data on serum and urine samples of clearance time 105-135 min showed a close correlation with inulin clearance (x) calculated from the data obtained by the anthrone method, for which aliquots of the same samples were used (y = 1.066 x -1.429 ml/min, r = 0.997, n = 15).


[A new analytical method for simultaneous measurement of iothalamate and iohexol].

Glomerular filtration rate (GFR) is believed to be the overall index of renal function, and the renal clearance of inulin (Cin) obtained during constant intravenous infusion has long been accepted as the gold standard of GFR measurement. Because of a number of technical difficulties inherent in the assay of inulin concentration in urine and plasma, its utility in clinical practice is limited. Iothalamate, urographic contrast medium, which behaves and is excreted in a fashion similar to inulin, has been accepted as a good filtration marker. We examined whether GFR could be estimated by ionic contrast media, "iothalamate" and non-ionic contrast media, "iohexol", when the high-performance liquid chromatography (HPLC) system was used as the analytic method. In our HPLC system, iothalamate and iohexol could be simultaneously determined. The assay results gave linearity within the wide range of concentrations tested. In seventeen subjects, the renal clearances of creatinine, iothalamate, and iohexol (Ccr, Ciot, and Cioh) were compared with that of inulin. The slopes with zero intercept of Ccr vs Cin, Ciot vs Cin, and Cioh vs Cin were 1.24 +/- 0.02, 1.06 +/- 0.02, and 0.83 +/- 0.02, respectively. In conclusion, an excellent correlation of renal clearance of iothalamate with that of inulin was obtained in patients with a wide range of renal function and normal volunteers. This justifies the use of the single injection of iothalamate and HPLC system as the analytic method in the measurement of GFR.


[Doppler ultrasound measurement of phasic renal arterial blood flow velocity in patients with chronic glomerulonephritis].

The phasic renal arterial blood flow velocity was measured using a Doppler-Based Toshiba SSH-160A scanner in 25 healthy subjects and 78 patients with chronic glomerulonephritis. Renal arterial blood flow at the renal hilum was visualized with color Doppler ultrasound, and the velocity waveform was obtained by pulsed Doppler ultrasound. The velocity waveform was then analyzed to give the peak systolic velocity (S), end-diastolic velocity (D), resistive index (RI), and pulsatility index (PI). Creatinine clearance correlated with S (r = 0.76), D (r = 0.80), RI (r = -0.74), and PI (r = -0.85). The split renal glomerular filtration rate, calculated by a method which makes use of the early renal uptake of Tc-99m DTPA, also correlated well with these parameters. These findings suggest that renal arterial blood flow as detected by Doppler ultrasound may be useful for the noninvasive, direct, rapid, and simple evaluation of renal hemodynamics and renal function, although various modifying factors also need to be considered.


[Clinical significance of urinary sediment dysmorphic red blood cells and casts in renal disease].

We investigated the utility of measuring dysmorphic red blood cells (d-RBC) and urinary casts in the diagnosis and assessment of prognosis of renal diseases by evaluating these parameters in a total of 278 urine specimens from 78 patients with various renal diseases at our hospital. Urinary sediments obtained by the conventional method with in 4 hours after voiding were subjected to Sternheimer and Malbin's staining and examined under a conventional light microscope. We used the criteria we previously devised for classification of RBC as d-RBC and for classification of casts hyaline, granular, epithelial, red blood cell, white blood cell, fatty, or waxy. When there were 5 or more urinary sediment red blood cells per high-power field, d-RBC was found in 94% of samples from patients with chronic glomerulonephritis. The finding of d-RBC was not related to glomerulonephritis pathologically diagnosed according to the WHO classification system for biopsy specimens. Even when there were fewer than 5 red blood cells per high-power field, d-RBC was sometimes found. Casts of all types were found in samples from patients with chronic glomerulonephritis or chronic renal failure. In samples from patients with chronic pyelonephritis, however, white blood cell, fatty, and waxy casts were not found, and less than 1 granular or epithelial cast per low-power field was found. The findings for the presence and the extent of both d-RBC and casts were in good accord with and changed with the clinical course, indicating that d-RBC and casts of urinary sediment can be used to predict the prognosis of patients with renal disease.


[Urinary cytology for carcinoma of the urinary bladder].

Cytologic examinations of voided urine were reviewed for 82 cases of invasive or noninvasive papillary carcinoma and 25 cases of carcinoma in situ of the urinary bladder. Five of the 82 cases of invasive or noninvasive papillary carcinoma and 3 of the 25 cases of carcinoma in situ were asymptomatic, and malignancy was detected only by urinary cytology. Urinary cytology was positive in 18.5% of the Grade 1, 33.3% of the Grade 2, 75% of the Grade 3 noninvasive carcinomas and in 77.8% of the invasive carcinomas. However, all 25 cases of carcinoma in situ gave positive cytologic results. Malignant cells found in Grade 1 noninvasive papillary carcinoma were slightly atypic in shape. Their nuclei were small but showed mild to moderate hyperchromasia. These malignant cells appeared in small clusters but the number of clusters were few. In Grade 2 or 3 noninvasive papillary carcinoma, malignant cells appeared in small clusters or isolated single cells. Their nuclei were irregular in shape and showed moderate to marked hyperchromasia. In invasive carcinoma and carcinoma in situ, many malignant cells with marked atypia were observed, but in carcinoma in situ, the background of the specimens was clean.


[Pathophysiological analysis of diabetes mellitus and complications from the urine of diabetic patients].

Of the many information obtainable from the urine of diabetic patients, urinary C-peptide (CPR), albumin and anti-diuretic hormone (ADH) were representatively described using my clinical and experimental data. C-peptide excretion in 24h collection of urine is a good estimate of insulin secretion from the pancreas and thus low in IDDM patients and even in NIDDM patients at a later stage, but high in pathological conditions including Graves' disease, obesity, liver cirrhosis and Cushing's syndrome. Urinary albumin excretion in small amounts (microalbuminuria) is usually observed in diabetic patients who have been under a poor control state of diabetic hyperglycemia for over 5 years and provides a good tool for monitoring early diabetic nephropathy. The grade of microalbuminuria (30-300 mg/day) is positively correlated with the HbA1 level in diabetic patients, showing that microalbuminuria is reversible along with an improvement of diabetic control at least in an early phase of diabetic nephropathy. As the albumin level measured in a spot urine sample correlates well with the value in the 24h collection of urine, the albumin measurement is conveniently feasible with a spot urine sample at every patient's visit. The amount of ADH excreted in urine is 7-10% of that secreted from the posterior pituitary. The excretion of ADH in a day was in the urine of diabetic patients positively correlated with HbA1, urinary osmolarity and concentration of sodium in urine, although the pathological meaning of the observed ADH hypersecretion in the development of diabetic complications is currently unknown.


[Quantitative analysis of urinary organic acid].

We showed the optimum conditions for analysis of urinary organic acids using solvent extraction and GC/MS by the stable isotope dilution technique. Examples of the diagnosis in a case of heterozygote of ornithine transcarbamylase deficiency and a case of multiple carboxylase deficiency were described. The accurate quantitative analysis of urinary organic acids with stable isotope dilution technique is necessary for routine examination conducted in clinical laboratories.


[Urinary trypsin inhibitor as an acute phase reactant].

Early in this century, trypsin inhibiting activity has already been recognized in patients with acute infection or renal disease. In addition to these, conditions such as coronary thrombosis, surgical operation, artificial fever by heat-killed bacilli, malignancy, leukemia, later stage of normal pregnancy, etc. have been known to cause the elevated excretion of UTI in urine. Typically, maximal excretion of UTI has been observed within one or two days after the onset. It appears that recent studies have overcome the complexity of UTI molecule. Automated measurement of urinary trypsin inhibitor (UTI) in urine sample was carried out by either enzymic or immunologic method. UTI as well as erythrocyte sedimentation rate and C-reactive protein enables us to monitor acute phase response, being confirmed in cases of abdominal surgery.


[Immunophenotypic and immunogenotypic analyses of acute leukemia and lymphoblastic lymphoma expressing myeloid and lymphoid antigens].

By using monoclonal antibodies against lymphoid and myeloid differentiation antigens, surface marker analysis was performed on the tumor cells from 42 patients with acute leukemia and lymphoblastic lymphoma. Nine (21%) of 42 cases were diagnosed biphenotypic leukemia. Two (17%) of the 12 patients with acute myeloid leukemia, four (18%) of 22 with acute lymphocytic leukemia and three (38%) of 8 with lymphoblastic lymphoma expressed both lymphoid and myeloid antigens. Tumor cells from six patients expressed both T-cell and myeloid antigens, and those from three other expressed both B-cell and myeloid antigens. Southern blot analysis was performed on the DNA from four patients with biphenotypic leukemia cells expressing T-cell and myeloid antigens. DNA from one patient showed clonal rearrangement of the immunoglobulin heavy chain (IgH) gene, and that from one other showed clonal rearrangement of both IgH gene and T-cell receptor beta-chain gene. DNA from two other patients showed a germline configuration of both genes. These results indicate that biphenotypic leukemia, especially T-cell and myeloid phenotype, is not so rare in acute leukemia and lymphoblastic lymphoma. The results of immunogenotypic analysis were not consistent with those of immunophenotypic analysis in biphenotypic leukemia.


[A method for determination of proinsulin levels in serum using both insolubilized anti-insulin antibody and anti-C-peptide antibody].

A method is described for determination of proinsulin levels in serum. The principle of the assay is that proinsulin reacts with both anti-insulin and anti-C-peptide antibodies. The assay procedure is as follows; Anti-insulin antibody fixed to bacterial cell wall and insolubilized is incubated with test serum to form a complex of proinsulin-anti-insulin antibody (solid phase), followed by washing twice with buffer to eliminate free C-peptide. Then, glycin-HCl buffer is added to dissociate the bound proinsulin. After centrifugation, the supernatant is neutralized with NaOH and proinsulin in it is measured using RIA kit for CPR assay. The assay is simple, sensitive and reproducible. Neither insulin nor C-peptide contained in test serum influences the proinsulin levels determined by this assay. The mean +/- S.D. of the fasting serum proinsulin levels of healthy donors was 7.0 +/- 2.6 PM/l. A patient with insulinoma showed extremely high serum proinsulin level, which decreased to the normal range after extirpation of insulinoma.


[Evaluation of serum troponin T measurement in acute myocardial infarction].

A monoclonal solid phase enzyme immunoassay has been developed for the detection of human troponin T. The serum troponin T levels in healthy subjects gave 0.05 +/- 0.06 ng/ml in total (n = 176), 0.06 +/- 0.07 ng/ml in males (n = 79) and 0.03 +/- 0.05 ng/ml in females. Within-run and between-run precision (CVs) of the assay were less than 5%. Various common interferents tested did not affect on the assay, but higher titer of rheumatoid factor, and anti-coagulants such as EDTA, heparin oxalate and citrate affected the assay. In all patients with defined acute myocardial infarction, serum troponin T levels increased 7 to 10 folds the upper reference range within 6 hours after the onset of chest pains and maximum elevation of serum troponin T level was at around 20 hours and its levels remained elevated for 7 to 20 days. Specificity and sensitivity for acute myocardial infarction was 92.4% and 100%, respectively. The results indicated that troponin T measurement improved the diagnostic efficiency for the detection of myocardial necrosis as compared with conventionally used cardiac enzymes and was an effective tool for the confirmation of the reperfusion by PTCA and PTCR.


[Study on changes of serum vitamin K1 level and K dependent coagulation factors in patients with coumarin derivatives (warfarin) therapy].

Serum level of vitamin K1 (= phylloquinone, hereinafter K1) and K dependent blood coagulation factors were determined by HPLC in normal subject, liver cirrhosis, hepatocellular carcinoma, acute hepatitis, chronic hepatitis, chronic renal failure with hemodialysis and patients under warfarin therapy. Normal range of serum K1 concentration was decided on 0.20-2.30 (0.87 +/- 0.53, n = 96) ng/ml. Serum K1 level showed no significant differences among normal subject, various diseases and warfarin therapy. Correlation between serum K1 level and F-VII (r = 0.879, p less than 0.001) or protein C activity (r = 0.839, p less than 0.01) was found in patients whose thrombotest was 20% and less. However serum K1 level didn't correlate with any K dependent coagulation factors in patients if thrombotest was over 20%.


[Differentiation of fibrinolysis and fibrinogenolysis by analysis of FDP fragments].

We previously analysed the fragments of fibrin/fibrinogen degradation products (FDP) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) combined with immunoblotting. In this report, we studied the semi-quantitative analysis of fibrinolysis (degradation of cross-linked fibrin) and fibrinogenolysis (degradation of fibrinogen and/or unstable fibrin) of patients' samples by our method. In vitro study of FDP made it clear that an appearance of D fragment confirmed fibrinogenolysis and an appearance of DD fragment and/or high molecular weight fragments which have higher molecular weight than DY or X fragment confirmed fibrinolysis. In addition, a study with mixtures of various concentrations of fibrin degradation products (FbDP) and fibrinogen degradation products (FgDP) demonstrated a dose dependent intensity of band by immunoblot method. These results show that our method is favorable for the semi-quantitative analysis of fibrinolysis and fibrinogenolysis. We applied the method to 6 samples from patients with disseminated intravascular coagulation (DIC). Consequently, fibrinogenolysis was observed in all of 6 samples, in which fibrinogenolysis was more enhanced than fibrinolysis in one sample, and an equivalent degree of fibrinolysis and fibrinogenolysis were observed in 3 of 6 samples. Although our method was probably devoid of the ability to distinguish FgDP from degradation products of unstable fibrin, these findings indicate that fibrinogenolysis is, at any rate, enhanced in the majority of patients with DIC, besides fibrinolysis.


[Immunohistochemical study of EGF and EGF-R on the esophageal carcinomas--from a standpoint of depth of invasion].

We performed immunohistochemical stainings for epidermal growth factor (EGF) and epidermal growth factor receptor (EGF-R) on 63 resected esophageal carcinomas without preoperative treatment and 12 cases with preoperative radiation to clarify a relationship between positivity and depth of invasion. EGF and EGF-R showed a similar positivity (75% of early cases and 88.9% of advanced ones invaded beyond submucosa). In advanced carcinomas, the positivity in each layer was 75% in the mucosa, 86.7% in the submucosa and muscle layer, and 93.3% in the adventitia. All lesions of nodal metastases were positive for these stainings. Sixty % of cases with preoperative radiation were positive. The degenerated cells showed weak positivity. However, the viable cells showed similar positivity to those of non-treated cases.


Gerontological education in Japan--in training of social welfare personnel and nurses.

Though Japan is noted for the custom of respect for the elderly, gerontological education is not widely practiced. At present it is only done in the training of several professionals and semi-professionals in health and welfare services for the elderly, i.e., medical doctors, nurses, social workers, and care workers for the disabled and older persons. In the training of social workers who are expected to serve as the pivot of a team for psycho-social help and caregiving, gerontological education is given as a part of social work education. In schools for care workers for the disabled and older persons that are two-year vocational schools, practical gerontological knowledge and skills needed in the care of the elderly are taught as an important base for their future work. In schools of nursing, most of which are two-year or three-year vocational schools in Japan, gerontological education is included as an indispensable component of the curriculum and is being given increasing importance these days.


Gerontological education in Japan--geriatric education and training.

There are 13 academic geriatric departments among 80 medical schools in Japan as of November 1991. The first independent department was established in 1962 at Tokyo University. The undergraduate education program includes lectures in geriatrics (20 hours/year in 11/12 medical schools), bedside teaching at geriatric ward (6/12 medical schools, 66 hours on average per year). The theme of lectures are diverse and incorporate all the three major fields in gerontology: biology of aging, clinical geriatrics and socio-economical aspects of aging society. The postgraduate geriatric education is carried out mainly at university setting and most of the medical schools (83%) accept graduate students who are trained at independent geriatric ward (92%) as well as at outpatient clinics. In 1989, Japan Geriatrics Society started a new certification system by which 687 MDs have been temporarily certified in geriatrics. The first examination will be given by the society in 1992 and the eligibility to sit in the examination requires three years geriatrics fellowship after certification in medicine or general surgery. The curriculum proposed by the society shares many items of training in common with those found in north American and in Europe. Some points of suggestions and recommendations were presented for future improvement in the education of gerontology in Japan.


International comparison in gerontological education--Canada.

The undergraduate programs of geriatric education in medical schools across Canada are variable, and there has not been definition of core content or core clinical experience, although these processes are underway. At UBC in June 1990, the first Canadian Summer Institute took place in Vancouver and medical students representing 16 medical schools in Canada attended the week-long program. Students left the program keen to start student interest groups in their schools. Many students were indicating commitment to careers in geriatrics, both clinical and research. There is a Certificate of Special competence by examination in Canada, administered by the Royal College of Physicians and Surgeons of Canada. The standards for the program have been in place since 1981, and since 1984 candidates for the examination must have a minimum of two years training in Geriatric Medicine after at least three years training in Internal Medicine. To date, there are less than 75 certified Specialists in Geriatric Medicine in the country. The College of Family Physicians of Canada encourages training in care of the elderly during the two-year training program in Family Practice. In some schools (e.g. UBC) at least one month is mandatory. There is a Joint Committee with representatives from the Royal College and the College of Family Physicians making recommendations for geriatric training in Family Practice programs in Canada. The College of Family Practice has elected not to have a program for certification in geriatrics for family physicians. Gerontology and geriatrics are being incorporated into undergraduate and graduate curriculae, driven by the demographic imperative. There remains much to be done.(ABSTRACT TRUNCATED AT 250 WORDS)


Gerontological education in Australia.

Courses in Gerontology and Geriatric medicine are now provided in most undergraduate training programs for medical, allied health and social work professionals in Australian Universities. The depth and coverage varies between schools but almost all undergraduate social workers, nurses, medical practitioners and allied health professionals will receive some training and in several cases more extensive optional programs are offered.


The growth of gerontology & geriatrics in the United States.

Since the confirmation of the Older American's Act (OAA) in 1965, the growth of gerontology and geriatrics, has literally come of age in the United States. Although individual aging courses were offered in higher education prior to passage of the OAA, few gerontology programs had been established. On campuses where gerontology courses were available, they represented electives connected with a pre-existing discipline, such as social work or family studies. However, 1965 was a watershed year for gerontology instruction since it heralded the provision of federal funding support, beginning with that of the Administration on Aging (AoA), to help develop, pilot, and oversee gerontological programs in American institutions of higher learning. Initially, instruction was at an undergraduate level, but later gerontology degree programs were established at the master's level, with the first gerontology degree program originating at North Texas State University, in 1967. This program was followed a year later (1968) by a program at the University of South Florida. While funding from AoA began in 1966, funding from other federal agencies (e.g., the Bureau of Health Professions (BHP); the National Institute on Aging (NIA); the National Institute of Mental Health (NIMH), Mental Disorders of Aging Branch; and the Veteran's Administration (VA))--the five agencies that became the basis for a 1984 and 1987 Report on Education and Training in Geriatrics and Gerontology,--did not begin until the mid 1970s. The mid 1970s reflected the growing awareness of the demographic shift in America and around the world, particularly in developed nations.(ABSTRACT TRUNCATED AT 250 WORDS)


[Acute hemodynamic effects of intravenous bolus injection of isosorbide dinitrate in aged patients with congestive heart failure].

In order to investigate whether intravenous bolus injection of isosorbide dinitrate (ISDN) is a safe and efficient therapy in aged patients with congestive heart failure, we studied acute hemodynamic effects in 11 patients. Peak effects on preload were observed after 5 to 10 minutes of bolus injection and unloading effects continued effectively for 60 minutes. At peak effect, pulmonary systolic pressure decreased from 50.2 +/- 2.6 to 36.2 +/- 2.6 mmHg (-28.5%, p less than 0.01) and pulmonary end diastolic pressure decreased from 25.0 +/- 2.2 to 18.5 +/- 2.1 mmHg (-26.0%, p less than 0.01). Mean pulmonary artery wedge pressure decreased from 23.4 +/- 2.2 to 16.0 +/- 2.1 mmHg (-31.6%, p less than 0.01). Mean right atrial pressure decreased from 10.5 +/- 1.8 to 7.4 +/- 2.0 mmHg (-29.5%, p less than 0.01). Blood pressure, heart rate, cardiac index, systemic and pulmonary vascular resistance showed no significant changes. Thus, intravenous bolus injection of ISDN showed a potent vasodilator effects on preload, and may be a safe and useful treatment for aged patients with acute congestive heart failure.


[Clinical significance of measuring glycated fibrinogen in plasma in diabetes mellitus].

The clinical significance of measuring glycated fibrinogen (G Fbg) in plasma was studied using a simple, rapid method for determining plasma G Fbg developed by the authors. This method is based on modified fibrin clot formation for purification of plasma Fbg, and G Fbg was estimated by improved colorimetric fructosamine assay. The values of plasma G Fbg were expressed as glycated polylysine equivalents (mumol/l). In this clinical study, 106 examinees who underwent a screening test for diabetes mellitus (DM) because of glycosuria or hyperglycemia were investigated. According to the WHO diagnostic criteria, they were divided into 3 groups: non DM group (n = 35), impaired glucose tolerance (IGT) group (n = 31) and DM (non-treated) group (n = 40). In addition, healthy controls (n = 40) and treated diabetic patients (DM treated) were involved. Plasma G Fbg/Fbg, fasting plasma glucose (FPG), fructosamine and glycated hemoglobin (HbA1c) were also analyzed in subjects of the 5 groups. Non-treated DM patients showed high levels of plasma G Fbg, which were significantly higher than those of any other groups. G Fbg levels in IGT group were significantly higher than those in healthy controls or the non-DM group. Together with a significant positive correlation between G Fbg and FPG, these results indicate that the levels of plasma G Fbg depend on plasma glucose levels.(ABSTRACT TRUNCATED AT 250 WORDS)


[The relation between bone calcium metabolism and senile aortic valve calcification].

The authors assessed the affect of bone calcium metabolism on aortic valve calcification (AVC) in 189 septua- and octogenarians (49 males and 140 females, 81.0 +/- 4.4 yrs). Both AVC and mitral annular calcification (MAC) were evaluated by two-dimensional echocardiography, and then the degree of AVC was classified into three categories; C-1: calcification seen in one cusp of the aortic valve, C-2: calcification in two cusps, and C-3: calcification in three cusps. Bone mineral content (BMC) of three lumbar vertebral bodies was obtained by quantitative computed tomography using a calibrated phantom. Serum calcium, phosphate, parathyroid hormone, calcitonin, and osteocalcin were also examined within a month. The patients were classified into age-matched five groups in both sexes; Group-C: MAC (-) and AVC (-) (n = 79); Group-A1: MAC (-) and AVC (+) with C-1 (n = 35); Group-A2: MAC (-) and AVC (+) with C-2 (n = 19); Group-A3: MAC (-) and AVC (+) with C-3 (n = 15); and Group-AM: MAC (+) and AVC (+) in any cusp (n = 42). In males, BMC decreased in the order of Groups-A3 (83 +/- 27 mg/cm3), -C (67 +/- 50), -AM (62 +/- 62), -A2 (61 +/- 38), and -A1 (59 +/- 58), but there was no significant difference between any of the five groups. In females, the BMC in Group-AM (29 +/- 24) was significantly less than that in Group-C (48 +/- 35) (p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)


[Bactericidal clothes protect against respiratory infections in the elderly patients].

In order to protect against infections in the elderly patients, bactericidal clothes were used. Ninety elderly patients were divided into two groups; the control group and the other group with bactericidal clothes. In period I neither group received particular treatment for three months and in period II only the treated group used bactericidal clothes for three months. The control group in period II used regular clothes throughout. Cause of fever (greater than 37 degrees C) extending more than 10 or 15 days were significantly lower in the treated group than in the control group. It was suggested that the bactericidal clothes reduce infections in elderly patients.


[An autopsy case of endocardial cushion defect (ECD) in an 82-year-old female].

In an 82-year-old female case of endocardial cushion defect (ECD), a systolic regurgitant murmur was heard at the apex, and her ECG showed atrial fibrillation without right bundle branch block or left axis deviation. An echocardiogram demonstrated atrial septal defect (ASD) and a cleft of the anterior mitral leaflet with calcification. She died of refractory congestive heart failure. Autopsy revealed ECD (intermediate type) with mitral and tricuspid cleft, and ASD (ostium primum type, 2.0 x 1.0 cm in diameter). In addition, mitral ring calcification and calcification of the cleft mitral valve was disclosed, causing mitral stenosis in addition to mitral regurgitation due to the cleft mitral valve. This was the second oldest Japanese autopsy case of ECD. We concluded that echocardiographic examinations, including color flow imaging, in aged patients with heart murmur are necessary to confirm the diagnosis of congenital heart disease in the aged.


Antiarrhythmic effects of MS-551, a new class III antiarrhythmic agent, on canine models of ventricular arrhythmia.

The antiarrhythmic effects of MS-551, which prolongs cardiac action potential duration without affecting the maximum upstroke velocity of the action potential, were assessed in three different canine ventricular arrhythmia models: 1) ventricular tachycardia (VT) induced by electrical stimuli 3-5 days after myocardial infarction, 2) spontaneous ventricular tachyarrhythmias 24-48 hr after two-stage coronary ligation and 3) ventricular tachyarrhythmias induced by digitalis. Intravenous administration of MS-551 (0.1-1 mg/kg) decreased the susceptibility in 10 dogs out of 13 to VT or ventricular fibrillation evoked by programmed electrical stimulation (PES) delivered to the ventricular septum 3-5 days after myocardial infarction. Oral administration of MS-551 (3 mg/kg) also decreased the susceptibility to VT evoked by PES in 7 out of 10 conscious postinfarction dogs. Concurrently, intravenous (0.1-1 mg/kg) or oral (3 mg/kg) administration of MS-551 produced increases in the ventricular effective refractory periods (ERP) by 7 +/- 1% - 17 +/- 3% or 13 +/- 2%, respectively. Similarly, d-sotalol (0.3-3 mg/kg, i.v. and 10 mg/kg, p.o.) decreased the susceptibility to VT with increased ERP. However, MS-551 (1 and 10 mg/kg, i.v.) failed to inhibit both canine two-stage coronary ligation arrhythmia and digitalis arrhythmia. These results suggest that MS-551 is a pure class III antiarrhythmic drug which may be effective in the treatment of life-threatening reentrant tachyarrhythmias, but not in automaticity arrhythmias.


Effect of nebracetam on the disruption of spatial cognition in rats.

Central cholinergic hypofunction causes the disruption of spatial cognition, while cholinomimetics improve this disruption in rats. Scopolamine (0.5 mg/kg, i.p.) has also been reported to disrupt radial maze performance in rats. Nebracetam (WEB 1881 FU), a new nootropic candidate, was able to correct this scopolamine-induced disruption of spatial cognition at the dose of 10 mg/kg, p.o. Furthermore, nebracetam enhanced oxotremorine-induced tremors in mice. These results indicate that nebracetam has a cholinergic enhancing effect. The scopolamine-induced disruption of spatial cognition has been previously reported to improve not only by cholinomimetics but also by brain noradrenergic drugs such as L-threo-DOPS and amantadine. Nebracetam reversed the change of brain noradrenaline contents in the frontal cortex and hippocampus in which the noradrenaline content decreased by treatment with scopolamine. Nebracetam also decreased the delta 9-tetrahydrocannabinol (6 mg/kg, i.p.)-induced disruption of spatial cognition, which was reported to be related to the lymbic noradrenergic function. These results suggest that the cognitive enhancing effect of nebracetam involves not only cholinergic mechanisms but also involves lymbic and hippocampal noradrenergic mechanisms.


The effects of oxiracetam (CT-848) on local cerebral glucose utilization after focal cerebral ischemia in rats.

The effects of oxiracetam on the reduction of brain metabolism induced by focal cerebral ischemia were investigated by measuring local cerebral glucose utilization (LCGU) in rats 24 hr after left middle cerebral artery occlusion. Focal cerebral ischemia reduced LCGU in the entire ipsilateral cortex, the greatest reduction being in the lateral parts of the frontoparietal cortex. LCGU was slightly reduced in the contralateral cortex; this reduction was considered to be caused by diaschisis. Oxiracetam was administered intraperitoneally for 3 days prior to middle cerebral artery occlusion. In the ipsilateral cortex, LCGU reduction was minimized in the ischemic center areas by oxiracetam at a dose of 400 mg/kg and in more extensive areas, by a dose of 800 mg/kg. Moreover, oxiracetam at a dose of 800 mg/kg enhanced metabolism impaired by diaschisis in the caudal areas of the contralateral cortex. These findings suggest that oxiracetam minimizes the reduction of brain function induced by ischemia and may therefore be useful in the treatment of cerebrovascular disease.


Effect of L-NG-nitro-arginine, inhibitor of nitric oxide synthesis, on autoregulation of renal blood flow in dogs.

The present experiments were designed to evaluate the importance of nitric oxide in the regulation of renal hemodynamics and the autoregulation of renal blood flow (RBF) in anesthetized dogs. RBF was measured by an electromagnetic flowmeter, and renal arterial pressure (RAP) was varied by an adjustable aortic clamp. The RAP-RBF relations were examined during the intrarenal infusion of saline or agents. The intrarenal infusion of L-NG-nitro-arginine (L-NNA, 40 micrograms/kg.min) at normal RAP decreased RBF and urine flow (UF), while the infusion of L-arginine.HCI (1 mg/kg.min) increased RBF and UF. Both agents did not affect the glomerular filtration rate and mean arterial pressure. The autoregulation of RBF was impaired during the L-NNA infusion. The L-arginine infusion did not affect autoregulatory efficiency. When L-NNA (40 micrograms/kg.min) and L-arginine were infused simultaneously into the renal artery, the autoregulation of RBF was maintained. However, a higher dose of L-NNA (200 micrograms/kg.min) impaired the autoregulation of RBF. These results suggest that the basal production and/or the release of nitric oxide contributes to the regulation of renal hemodynamics and urine formation. During the reduction of RAP, nitric oxide may play an important role in the autoregulation of RBF.


Effects of sulfhydryl-related compounds on indomethacin-induced gastric lesions in rats: role of endogenous sulfhydryls in the pathogenesis.

The role of mucosal sulfhydryl (SH) in the pathogenesis of indomethacin-induced gastric lesions was investigated in rats. Indomethacin (25 mg/kg, s.c.) caused high-amplitude gastric contractions, resulting in linear hemorrhagic lesions in the corpus mucosa within 4 hr, but did not induce any changes in the mucosal SH levels. These lesions were prevented significantly by prior s.c. administration of cysteamine, glutathione (GSH) or diethylmaleate (DEM), irrespective of whether the mucosal SH was increased by the former two agents or reduced by the latter. N-Ethylmaleimide (NEM) tended to worsen such lesions without any effect on the mucosal SH contents. Gastric hypermotility caused by indomethacin was inhibited significantly by DEM, cysteamine and GSH, while acid secretion was reduced by DEM and NEM. Both cysteamine and GSH prevented the indomethacin-induced linear lesions even in the stomach perfused with 150 mM HCl, whereas in the animals treated with DEM, nonlinear damage was induced exclusively in the antrum by indomethacin in the presence of acid. We conclude that the mucosal SH has no relation to the ulcerogenicity of indomethacin in the gastric corpus mucosa.


Consecutive treatment of disulfiram inhibits ovarian carbonyl reductase activity in rats.

We investigated the effects of disulfiram (DS) on ovarian carbonyl reductase activity in rats to determine the influence of DS on female reproductive function. Three consecutive treatments with DS significantly inhibited ovarian carbonyl reductase activity as well as ovulation, dose-dependently. Single treatment with DS had no effect on ovarian carbonyl reductase activity. Our observations indicate that consecutive treatment with DS has an inhibitory action on female reproductive function, although DS is well-known to inhibit aldehyde dehydrogenase and dopamine beta-hydroxylase.


Working and reference memory in rats in the three-panel runway task following dorsal hippocampal lesions.

Using a three-panel runway task, the influence of dorsal hippocampal lesions on working and reference memory in rats was investigated. Despite 20 postoperative training sessions, rats with hippocampal lesions were unable to perform the working memory task. In the acquisition process of the reference memory task, however, there was no significant difference between hippocampal- and sham-lesioned rats. On the other hand, rats trained preoperatively with a working memory procedure, and then subjected to hippocampal lesions, showed more errors (pushes made on the two incorrect panels of the three panel-gates located at four choice points) than did sham-lesioned rats. The increase in working errors induced by hippocampal lesions was not reduced during 10 subsequent re-training sessions. Hippocampal lesions had no effect on retention of the reference memory performance. The increase in working errors in hippocampal-lesioned rats was significantly reduced by treatment with the cholinesterase inhibitor physostigmine at 0.1 mg/kg and the cholinergic activating drug minaprine at 10 mg/kg. These findings suggest that lesions of the dorsal hippocampus selectively impair the ability to carry out the working memory task whether rats are trained preoperatively or postoperatively, and that the working memory loss in hippocampal-lesioned rats is mediated by lowering of the cholinergic function.


Accelerating action of quinidine on the decay phase of transient outward current in dissociated hippocampal pyramidal neurons of rats.

The effect of quinidine on the transient outward current (IA) in rat hippocampal neurons was studied. Quinidine at 1 x 10(-6) M or more dose-dependently decreased the IA without affecting the current-voltage relationship. Quinidine accelerated the decay phase of the IA, suggesting the blockade of open channels. This result may have an implication on the side effects of quinidine in the central nervous system, since the IA is involved in the control of rhythmic electrical activity of the neurons.


Agonistic-antagonistic actions of clomiphene citrate on PGF2 alpha fluctuations in ovariectomized rat uterus.

Effects of clomiphene and/or estrogen on uterine prostaglandin (PG) F2 alpha fluctuations in ovariectomized rats were examined. Uterine PGF2 alpha fluctuations were measured at the indicated times after injection of clomiphene and/or estradiol. Antiestrogenic effects of clomiphene on urine weight and PGF2 alpha levels were observed at 6 hr, and its effect on uterine 13,14-dihydro-15-keto-PGF2 alpha forming capacity was observed at 24 hr. At 48 and 72 hr, additive effects were recognized in all parameters. These results indicate that clomiphene possesses agonistic-antagonistic actions on the PGF2 alpha fluctuations in the uterus.


Molecular orbital studies on the structure-activity relationships of catechol O-methyltransferase inhibitors.

Quantum chemical studies were applied to analyze the activities of catechol O-methyltransferase (COMT) inhibitors. Molecular orbital calculations of inhibitor molecules were made by semi-empirical molecular orbital calculations, CNDO/2 (complete neglect of differential overlap) methods. Regression analysis among theoretical reaction indices based on the frontier electron theory and COMT inhibitory activities were carried out. The COMT inhibitory actions of two series of inhibitors, a series of 1,5-substituted 3,4-dihydroxy benzenes and a series of substituted 3-hydroxy-4-methoxy benzenes, were investigated. The resulting regression equations contain two common reaction indices as regression variables: the electron density on the oxygen atom of the hydroxyl group and the super-delocalizability on the 5th carbon atom of the benzene ring. These two atomic positions are considered to play an important role in the interaction of these inhibitors with COMT. The hydroxyl of atomic position 3 is probably indispensable to the COMT inhibitory action by these inhibitors.


Neural and metabolic control of arteriolar diameter.

During sympathetic nerve activity, sympathetic transmitter causes depolarizations which trigger voltage dependent Ca++ entry. The ability of arteriolar membranes to sum together successive e.j.ps is changed as a consequence of cellular activity, i.e. increased [K+]o. Furthermore, the ability of arteriolar membrane to permit voltage dependent Ca++ entry is modified by a second consequence of cellular activity, i.e. changes in external pH.


Identification of a specific binding site for K+ channel openers in rat aorta.

The K+ channel openers (KCOs) form a structurally heterogeneous group of compounds which relax smooth muscle by opening K+ channels in the plasmalemma. At present it is not known whether these drugs open the same K+ channel in smooth muscle and, if so, whether they bind to the same site of this channel. To address these questions we present the first binding study with KCOs in a smooth muscle preparation. In intact rat aortic strips, the novel tritiated KCO, 3H-P1075 (N-cyano-N'-(1,1-dimethyl[2,2,3,3(3)H]propyl)-N"-3-pyridylguanidine++ +), a potent pinacidil analogue, showed saturable specific binding of high affinity (KD = 6 +/- 1 nM, Bmax = 23 +/- 3 fmol/mg tissue wet weight). Specific binding of 3H-P1075 was inhibited stereospecifically by representatives from all major families of K+ channel openers, with a rank order of potency that correlated well with the potencies for vasorelaxation in rat aorta. The sulfonylurea glibenclamide, a relatively specific blocker of ATP-sensitive K+ channels and an inhibitor of the effects of KCOs, also inhibited 3H-P1075 binding and increased the rate of dissociation of 3H-P1075 from the tissue in a concentration-dependent manner. Lowering temperature (from 37 degrees C to 2 degrees C) and decreasing intracellular ATP levels by metabolic poisoning, diminished specific 3H-P1075 binding by reducing Bmax. However, depolarization (KCl = 55 mM) or lowering pH from 7.4 to 6.0 did not influence binding. The data demonstrate the existence of a specific binding site for the KCO 3H-P10756 in rat isolated aorta, which seems to be of functional relevance.(ABSTRACT TRUNCATED AT 250 WORDS)


The role of intestinal bacteria in the transformation of sodium picosulfate.

Sodium picosulfate, a laxative, was biotransformed to 4,4'-dihydroxydiphenyl-(2 pyridyl)-methane by intestinal flora that produced a novel sulfotransferase (not sulfatase). The biotransformation was activated by adding phenolic compounds such as phenol, acetaminophen and flavonoids. The enzyme activity related to this biotransformation was the highest in the contents of the caecum region of the intestine. The enzyme activity was 3.0 mumole/hr/g wet feces in humans and 0.75 in rats (pH 8.0). The optimal pH was 9.0.


Effects of psychoactive drugs on short-term memory in rats and rhesus monkeys.

To examine the effects of drugs on short-term memory in animals, the delayed discrimination experiment in rats and the delayed matching to sample experiment in rhesus monkeys were conducted. Nicotine at 0.125 mg/kg, s.c. in rats and at 0.5 mg/kg, s.c. in monkeys increased the percentages of correct choices. Scopolamine at 0.06-0.12 mg/kg, s.c. in rats and at 0.015 mg/kg, s.c. in monkeys decreased the percentages of correct choices. However, supposedly memory-specific, delay-time-dependent disruptive effects by scopolamine were found only in monkeys. Diazepam at 0.5-2 mg/kg, s.c. did not change the correct choices in rats. However, diazepam at 1-4 mg/kg, i.g. decreased the correct choices in monkeys regardless of the delay time. Chlorpromazine at 0.25-1.5 mg/kg, s.c. showed inconsistent effects in rats. In monkeys, chlorpromazine at 0.25-0.5 mg/kg, s.c. had no effect. These results suggested that using both rats and monkeys would be useful for evaluating the effects of drugs on memory.


Chronopharmacology of probucol in mice.

Mice were maintained under conditions of light from 7 a.m. to 7 p.m. and dark from 7 p.m. to 7 a.m. Probucol was given orally to these animals once daily at 10 a.m. or 10 p.m. for 7 days. Blood samples for serum cholesterol were obtained at 24 hours after the final dosage. Blood samples for plasma probucol were obtained just before and at 3, 6, 12, 24, 48, 72, 96 and 120 hours after the final dosage. The cholesterol lowering effect of the agent at 10 p.m. was greater than that at 10 a.m. Plasma probucol concentrations of the two trials did not differ at any observation point. These data suggest that the effect of probucol varies with its time of administration. This might not be caused by a time-dependent change in plasma probucol concentration.


Cholecystokinin protects cholinergic neurons against basal forebrain lesion.

Alzheimer's Disease (AD) patients have a severe degeneration of cholinergic neurons in their cerebral cortices. Basal forebrain (BF)-lesioned rat is used as a model animal of a cholinergic deficit in the cerebral cortex. Cholinergic markers were decreased in the cerebral cortex of BF-lesioned rats. Intracerebroventricular continuous infusion of cholecystokinin octapeptide (CCK8) following BF lesion obviously preserved these cholinergic markers. These results suggest that CCK8 prevents the degeneration of cholinergic neurons in the cerebral cortex following BF lesion.


Effects of anti-asthma drugs and potassium channel openers on neurally-mediated contraction of isolated guinea pig trachea.

The effects of anti-asthma drugs, isoproterenol, aminophylline and hydrocortisone, and potassium channel openers on the contraction induced by electrical stimulation or exogenously applied acetylcholine were investigated in isolated guinea pig trachea. Isoproterenol and aminophylline non-selectively inhibited both the contraction evoked by vagus nerve- and that by transmural field-stimulation, but had no effect on the response induced by exogenously applied acetylcholine. Hydrocortisone and potassium channel openers, NIP-121 and cromakalim, preferentially inhibited vagus nerve-mediated response. These results suggest that anti-asthma drugs may have an ability to inhibit neurally-mediated contraction in the guinea pig trachea.


Protective effects of benidipine on arachidonic acid-induced acute cerebral ischemia in rats.

Acute cerebral ischemia was produced in rats by injection of arachidonic acid (AA) into the internal carotid artery. Evans blue (EB) was intravenously injected and its extravasation into the brain was determined as an indicator of disturbances in the blood-brain barrier and endothelial cells. Control animals showed severe cerebral edema and marked blue staining of the brain. Benidipine (30 micrograms/kg, i.p.) suppressed the increase in cerebral water content and the extravasation of EB. Similarly nicardipine (100 micrograms/kg, i.p.) suppressed the elevation of water content and the extravasation of EB. Furthermore, both benidipine (30 micrograms/kg, i.p.) and nicardipine (100 micrograms/kg, i.p.) improved the neuronal injuries following AA-injection. An antiplatelet agent, ticlopidine (100 mg/kg, i.p.), and a thromboxane A2 synthetase inhibitor, OKY-1581 (3 mg/kg, i.p.), also suppressed the elevation of cerebral water content. A lipoxygenase inhibitor, AA-561 (200 mg/kg, p.o.), and a cyclooxygenase inhibitor, indomethacin (10 mg/kg, i.p.), did not prevent the increase in cerebral water content. Neither benidipine (3-30 micrograms/kg, i.v.) nor nicardipine (100 micrograms/kg, i.v.) inhibited the AgNO3-induced thrombus formation of the abdominal aorta, whereas ticlopidine (100 mg/kg, p.o.) and OKY-1581 (3 mg/kg, i.v.) prevented the thrombus formation. From the present results, it is suggested that benidipine, as well as nicardipine, may protect against AA-induced acute cerebral infarction via a mechanism independent of antithrombotic action.


Beneficial effect of intravenous taurine infusion on electroretinographic disorder in taurine deficient rats.

We investigated the effect of intravenous taurine infusion on the electroretinogram (ERG) of taurine-deficient rats produced by treatment with guanidinoethyl sulfonate (GES), a taurine transport inhibitor. Mother rats were fed a taurine-free diet and given drinking water containing 1% GES from 2 weeks of gestation to weaning. The same feeding conditions were applied to male offspring after weaning. Both ERG measurement and continuous infusion of taurine at a dose of 10, 30 or 100 mg/animal/day were performed for 3 weeks from 7 to 10 weeks of age. GES-treatment reduced a- and b-wave amplitudes to 50% of the control levels and also increased b-wave latencies. Intravenous infusion of taurine improved these ERG abnormalities in a dose-dependent manner. Taurine concentrations in plasma, eyes and brain were also decreased by treatment with GES, and dose-dependent recovery was observed after infusion with taurine, although the concentrations of other amino acids were not affected by GES-treatment and infusion of taurine. Observations of morphological changes revealed that the retinal damage in GES-treated animals was decreased by taurine infusion. These results indicate that the changes in ERG and retinal structure observed in taurine deficiency are improved by intravenous infusion of taurine.


The role of endothelium in the phenylephrine-induced oscillatory responses of rabbit mesenteric arteries.

Phenylephrine-induced oscillatory contractions in rabbit mesenteric arteries were investigated in vitro. Adrenergic, cholinergic, or histamine antagonists as well as cyclooxygenase and lipoxygenase inhibitors had no effect on this phenylephrine-induced oscillation. The removal of extracellular calcium ions or treatment with a calcium antagonist reduced the amplitude and frequency of the oscillation. Removal of the endothelium or treatment with inhibitors of the synthesis or the target enzyme of endothelium-derived relaxing factor (EDRF) also reduced the amplitude and frequency of the oscillation. In a perfusion bioassay, the perfusate from an endothelium-intact arterial segment induced oscillation of an endothelium-denuded arterial ring recipient. These results suggest that phenylephrine-induced oscillation is mediated by an endothelium-derived factor such as EDRF and depends on the influx of extracellular calcium ions.


Platelet-derived growth factor (PDGF) accelerates induction of competence, and heparin does not inhibit PDGF-induced competence in primary cultured smooth muscle cells of rat aorta.

The time-dependent effect of platelet-derived growth factor (PDGF) on cell proliferation was investigated to clarify whether PDGF accelerates the rate of proliferation or its start in primary cultured smooth muscle cells (SMC) of rat aorta. In synchronized SMC at the G0 phase, 1, 10 and 100 ng/ml PDGF started DNA synthesis at 24, 15-18 and 12 hr, respectively, after stimulation by 3% fetal bovine serum (FBS) or hypophysectomized rat plasma (deficient in insulin-like growth factor-I (IGF-I)). Heparin (1, 10 or 100 micrograms/ml) decreased only the rate of DNA synthesis stimulated by PDGF in synchronized SMC. DNA synthesis in non-synchronized cells stimulated by PDGF with FBS was determined up to 10 days in culture. The stimulation with 1% FBS plus 30 ng/ml PDGF potentiated the DNA synthesis which was saturated with stimulation by 10% FBS alone, suggesting that prolonged treatment of PDGF transforms SMC. These results demonstrated that PDGF concentration-dependently accelerated the induction of competence independently of IGF-I, and heparin did not inhibit PDGF-induced competence but inhibited progression in primary cultured SMC of rat aorta.


Inhibitory effect of OP-41483.alpha-CD, a prostacyclin analog, on peripheral vascular lesion models in rats.

The effect of a chemically stable prostacyclin analog, OP-41483 alpha-cyclodextrin clathrate (OP-41483.alpha-CD), on vascular lesions, platelet aggregation and blood pressure were examined and compared with those of prostaglandin E1 alpha-cyclodextrin clathrate (PGE1.CD) in in vivo rat models. 1) In the laurate (1 mg/leg, i.a.)-induced arterial thrombotic model, OP-41483.alpha-CD (1 microgram/kg/min, i.v.) prevented the progression of femoral arterial vascular lesions and enhanced the development of collaterals in the femoral artery. PGE1.CD did not inhibit the progression of vascular damages. 2) In the model of vasoconstriction induced by epinephrine (0.05 mg/tail, s.c.) and ergotamine (2 mg/kg, s.c.), OP-41483.alpha-CD and PGE1.CD, at 1 microgram/kg/min, inhibited the progress of of tail gangrene and lessened the decrease in tail cutaneous blood flow. 3) OP-41483.alpha-CD (1 microgram/kg/min) suppressed the ADP (0.1 mg/kg/min, i.v.)-induced decrease in the number of circulating platelets without affecting the change in blood pressure. In contrast, PGE1.CD (3 micrograms/kg/min) inhibited ADP-induced thrombocytopenia with a decrease in blood pressure. These results indicate that OP-41483.alpha-CD has antiplatelet and cutaneous blood flow improving activities that are greater than its hypotensive effect and may be of therapeutic potential in peripheral vascular diseases.


Effects of pravastatin sodium alone and in combination with cholestyramine on hepatic, intestinal and adrenal low density lipoprotein receptors in homozygous Watanabe heritable hyperlipidemic rabbits.

Pravastatin sodium (pravastatin), a tissue-selective inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, was administered alone (50 mg/kg) or in combination with cholestyramine, a bile acid sequestrant resin, at the level of 2% in the diet to homozygous Watanabe heritable hyperlipidemic (WHHL) rabbits for 4 weeks. The low density lipoprotein (LDL)-cholesterol levels were reduced by 29% and 56% with pravastatin alone and the combination treatment, respectively. Hepatic LDL receptor activity was increased by 11.2- and 13.9-fold with pravastatin alone and the combination treatment, respectively. The LDL receptor activity in the untreated homozygous WHHL rabbits was only 2.5% of that in the normal rabbits. mRNA for the LDL receptor in the liver was also increased by 2.1- and 3.4-fold with pravastatin alone and the combination treatment, respectively. On the other hand, mRNA for the LDL receptor in the adrenal gland was not affected by pravastatin and the combination treatment, whereas the mRNA in the intestine was increased in both groups. These results suggest the following: 1) the induction of hepatic LDL receptor activity by the treatment of pravastatin alone or in combination with cholestyramine is the main cause of the reduction of serum cholesterol levels by these treatments even in LDL receptor-deficient animals. 2) The induction of the mRNA for the LDL receptor in the liver and intestine, but not that in the adrenal gland, might be a reflection of the tissue-selective inhibition of cholesterol synthesis by pravastatin.


Basic fibroblast growth factor ameliorates learning deficits in basal forebrain-lesioned mice.

The effect of basic fibroblast growth factor (bFGF) treatment on memory and learning performance ability was investigated in basal forebrain (BF)-lesioned mice. Eight-week-old male ddY mice underwent bilateral BF lesions by delivery of radiofrequency current. Basic FGF (5 or 50 ng/side) was microinjected into the same location immediately after lesioning. From fifteen days after the treatment, a step-through type passive avoidance test was performed daily for 10 days. Lesioned animals showed severe impairment in the acquisition process in this task, but not in the retention process. Basic FGF improved the step-through performances; step-through latency was elongated in a dose-dependent manner on the first test trial day and the mean time required to reach the acquisition criterion was shorter than in the vehicle-treated control group. However, bFGF did not alter the cortical choline acetyltransferase (ChAT) activity decrement induced by BF lesion. These results suggest that bFGF ameliorates the memory deficit without affecting the cortical ChAT activity.


Effect of pentazocine on the cytotoxicity of cortisone-resistant lymphocytes from mouse thymus.

Pentazocine and its related compounds were examined for their effect on the cytotoxicity of cortisone-resistant lymphocytes (CR lymphocytes) against Ehrlich carcinoma cells. The following compounds were used: pentazocine, naloxone, levallorphan, eptazocine and morphine. CR lymphocytes were obtained from the thymus or spleens of mice injected i.p. with hydrocortisone acetate (125 mg/kg) 2 days before harvesting the lymphocytes. The mixture of tumor cells and CR lymphocytes was inoculated s.c. into mice after incubation in the presence or absence of 10 microM drugs. Five weeks after inoculation, the percentage of mice developing a solid tumor among the recipients given the pentazocine-treated cell mixture of tumor cells and thymic CR lymphocytes was significantly smaller than the percentage in recipients given the cell mixture treated with or without other drugs (percent tumor takes: 21% and about 80%, respectively). Splenic CR lymphocytes did not show any cytotoxic effect, irrespective of the drug treatment. The pretreatment of CR lymphocytes or Ehrlich cells with 10 microM pentazocine did not affect the cytotoxicity of thymic and splenic CR lymphocytes. The proportion of the lymphocyte-conjugated tumor cells was significantly increased when the mixture of CR lymphocytes and tumor cells was incubated in the presence of pentazocine. The present results indicate that the cytotoxicity of thymic CR lymphocytes is enhanced by pentazocine possibly through the increase in the proportion of the lymphocyte-conjugated tumor cells but enhanced not by the other drugs.


Studies on antinephritic effects of plant components (3): Effect of pachyman, a main component of Poria cocos Wolf on original-type anti-GBM nephritis in rats and its mechanisms.

The antinephritic effect of pachyman on original-type anti-GBM nephritis in rats was investigated. Pachyman was given to original-type anti-GBM nephritic rats for 10 days from the day of anti-GBM serum injection. Pachyman prevented urinary protein excretion and the elevation of serum cholesterol content. Histopathological observations of the glomeruli indicated that although the number of nuclei and adhesion to capillary walls of Bowman's capsule in nephritic control rats were significantly increased, pachyman reduced the degree of histopathological changes such as hypercellularity and adhesion as compared to the control group. Although the serum complement CH50 ratio in control group was significantly lower than that in the normal group, the decrease in serum complement CH50 was inhibited by pachyman, and rat C3 deposition in the glomeruli in the pachyman-treated group was significantly reduced. These results suggest that pachyman was effective against original-type anti-GBM nephritis in rats and that the antinephritic mechanisms of pachyman may be partly due to the inhibitory action of this agent on C3 deposition in the glomeruli.


Ca2+ ionophore and phorbol ester stimulate diacylglycerol formation and phosphatidylcholine hydrolysis in rat parotid acinar cells.

We investigated the effects of A23187 and phorbol 12,13-dibutyrate (PDBu) on sn-1,2-diacylglycerol (DAG) accumulation and phosphatidylcholine (PC) hydrolysis in rat parotid acinar cells. Both A23187 and PDBu, in concentration ranges of 0.001-0.1 microM, stimulated DAG accumulation and PC hydrolysis in a time- and concentration-dependent manner. Treatment with A23187 and PDBu stimulated the release of [3H]choline and [3H]phosphocholine into the medium, indicating [3H]PC hydrolysis is due to the activation of phospholipases C and D; however, [3H]phosphatidylethanolamine hydrolysis was not indicated. These releases were unaffected by the addition of glucose 6-phosphate, a phosphatase inhibitor. Staurosporine, a protein kinase C inhibitor, significantly inhibited the DAG accumulation and the PC hydrolysis stimulated by these agents. Combinations of A23187 and PDBu potentiated the stimulatory effect which each of these agents alone had on DAG accumulation and PC hydrolysis. This mode of action was additive but not synergistic. These results suggest that DAG accumulation induced by A23187 and PDBu is related to the PC hydrolysis mediated via the activation of phospholipases C and D, and that it is not related to phosphatidylethanolamine hydrolysis.


[Itch and the heat tolerance scale: effects of thermographic biofeedback].

Effects of heat tolerance (HT) relating with individual emotionality, and of thermal biofeedback on itching were examined. Fourteen high HT subjects and 12 low HT subjects were selected by the Cold and Heat Tolerance Scales (Dienstbier, LaGuardia, & Wilcox, 1987). The experiment was divided into three phases: rating of itch, eight thermal biofeedback (increase or decrease) training sessions, and rating of itch with biofeedback. Itching was produced by a glue made from Japanese yam powder. Yam glue was applied on the subjects' dorsal forearm. The biofeedback information was given by the thermographics on the CRT, where its color was changed according to the changes in temperature of the dorsal forearm skin. Results showed that (a) subjects who were instructed to decrease their skin temperature reported comparatively lower intensities of itching, although their actual temperature controls were unsuccessful, and (b) the biofeedback performance reduced the intensity of itching for the low HT subjects, but raised it for the high HT subjects who were instructed to increase their skin temperature. The results suggested that the biofeedback performance could have different central effects on itching among the HT groups.


[The effect of motivation on preschool children's sharing judgment and behavior].

The purpose of this study was to examine the effect of motivation on preschool children's sharing judgment and behavior. In Experiment 1, 45 4-year-olds and 41 5-year-olds were asked whether they wanted to share seals in a fictional situation, and were observed whether they shared their seals in an actual situation. The main results indicated that the number of children who shared seals in the latter situation was smaller than those who judged to share seals in the former one. Especially there was a great difference regarding this point in 4-year-olds children. "Empathic orientation" motivated preschool children's sharing behavior, while "self-focused orientation" did not. In Experiment 2, 52 4-year-olds and 60 5-year-olds were divided into two empathic levels (middle, high) and two motivational cost levels (low, high) to examine the effects of "empathic orientation" and "self-focused orientation." The main results indicated that high motivational cost restrained children from sharing in an actual situation.


[Construction of a multiple mood scale].

A self-report instrument to measure multiple mood states was constructed. Six hundred and forty-eight Japanese affect adjectives were collected and divided into two lists. Japanese undergraduates totaling 1,254 rated each adjective in the list on a 4-point intensity scale to assess their momentary mood states. Factor analyses revealed 131 items with high loading on extracted major factors. The new list of these 131 items was rated three times by undergraduates totaling 1,768 and was factor analysed. Throughout the factor analyses, eight factors emerged with consistent reliability. Finally, 80 items highly loaded on these factors were selected to construct eight 10-item mood scales. These scales were called Depression (Anxiety), Hostility, Boredom, Liveliness, Well Being, Friendliness, Concentration and Startle. The scales were shown to be internally consistent. A comparison of these scales with the English mood scales was presented.


[Formation of grand coalition when consciousness of intragroup competition is activated].

The purpose was to explore conditions encouraging to form grand coalition, which leads a group to high solidarity with strong consciousness of intragroup competition. Subjects, 64 male undergraduates, were divided into 16 groups of four persons each. Experiment was carried out with 2 x 2 design of (consciousness of intergroup competition: strong vs. weak) (consciousness of intragroup competition: strong vs. weak). Results showed that strong competitive spirits against other groups produced facilitative effects on formation of grand coalition even when the consciousness of intragroup competition was strong, and the facilitative effects were strong even when consciousness of intragroup competition was weak. In addition, the necessary conditions for group members to form grand coalition under strong intragroup competition were discussed.


[Preschool children's prosocial judgments and their reasoning in the empathic situations].

The purpose of this study was to examine preschool children's prosocial judgments and their reasoning for prosocial episodes. One hundred children were individually asked to do prosocial judgments and their reasoning for three prosocial episodes: helping, sharing, and comforting. In the episode there was a crying or a normal facial expression of a person in distress under the condition of high and low empathic situations. The results indicated that the crying face and high empathic situation increased the rate of prosocial judgment more than the normal face and low empathic situation, respectively, and older children (5-year-olds) did prosocial judgment than younger ones (4-year-olds). Furthermore, the crying face induced the empathic reasoning, especially in the high empathic situation. These results supported that preschool children were able to use the empathic reasoning reflected in the negative facial expression and the cause of distress of the victim.


[A mediating role of physiological arousal in social facilitation].

An experiment was conducted to examine the mediating role of physiological arousal in social facilitation. It was hypothesized that the elevation of arousal level by the presence of other persons or evaluative apprehension would facilitate task performance. Twenty-four male and 24 female college students performed simple task alone, with a cooperative person, or with a competitive person. One half of the subjects was given an instruction which would produce evaluative apprehension. Subjects' skin potential responses were measured as the indices of physiological arousal. Although the hypothesis was supported by the analysis of psycho-physiological index, but it was not at all by the self-reported index of arousal.


[Conditioned aversion to an intraperitoneally injected odor CS in rats].

Two experiments were conducted to specify an CS property of intraperitoneally injected odor substance in conditioned odor aversion formation. In experiment 1, three groups of water-restricted rats received one trial of conditioning with intraperitoneal injections of orange-extract (CS) and LiCl(UCS). The CS-UCS intervals of experimental groups were 30 minutes (Group IE-30) and 120 minutes (Group IE-120). In the test trial, Group IE-30 rats showed aversion to the test solution (0.5% orange extract), whereas no significant difference was observed between the control group and Group IE-120. In experiment 2, three groups of rats received one or two or three conditioning trials (Groups E1, E2 and E3, respectively). The CS-UCS interval of all groups was 30 minutes. In the test trial, rats in Groups E2 and E3 consumed less amount of test solution (0.5% orange extract) than the control rats, whereas no significant difference was observed between Group E1 and the control group. These results suggest that the intraperitoneally injected odor substance have CS property on conditioned odor aversion.


[Effects of inclination of trunk and head on emotional awareness].

Posture is a nonverbal behavior and a universal means of animal and human communication. It is observed not only in interpersonal communication but in clinical situation. Our recent research shows that posture affects the mood and emotional awareness of the subjects. This study compared the subjects' awareness between two groups, the operational group (actual posture) and the image group, which only imagined a postural change. Six kinds of posture were adopted. These postures included two dimensions; inclination of trunk (straight or bent), and head (up, front or down). In these conditions the subjects estimated their mood and emotional awareness with 17 pairs of adjectives on a 3 point scale. The results of ANOVA showed statistically significant differences in conditions of both inclination of trunk and head. Especially, when the subject bent his back while hanging his head, this made most feeble, lifeless, and shadowy mood than any other postures. The authors confirmed that posture exerted a strong influence on one's emotional awareness.


[The current status of psychology in Korea].

This paper read as the special lecture at the Annual Convention of The Japanese Psychological Association, held at Tokyo Metropolitan University in 1990, and reported on the recent trends of psychology in the Republic of Korea. This report dealt essentially with four topics; 1) the history of Korean psychology which was divided into four periods of creation, reconstruction, development, and current status, after the liberation at the end of the World War II in 1945, 2) the distribution and academic status of 26 universities, with formal department of psychology, 3) general situation of job opportunities for the graduates in psychology of these universities, 4) the composition of Korean Psychological Association (KPA), consisting of nine divisions (social, industrial and organizational, clinical, consultations and psychotherapy, experimental and cognitive, developmental, biological and physiological psychology) with the total of about 400 members. Last, some directions of Korean psychology to the future was suggested.