Okayama Igakkai Zasshi (Journal of Okayama Medical Association)

Purification and characterization of HBs antigen from hepatoma huGK-14 cell line

Munehiro ODA

1989 Volume 101 Issue 7-8 Pages 687-698
Published: August 31, 1989
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.101.7-8_687

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HBs antigen was purified from the culture fluid of hepatoma huGK-14 cell line and its physico-chemical properties were studied.
The purification consists of following steps: concentration of culture fluid by membrane filtration, affinity column chromatography (anti-HBs monoclonal antibody column and anti-human serum albumin antibody column), and ultracentrifugation (isopycnic centrifugation in CsCl density gradient and rate zonal centrifugation on sucrose gradient). Highly purified (purity>99%) HBs antigen was isolated with an overall yield of about 40%. The HBs antigen showed uniform spherical particles (diameter: 23.2±2.9nm) and had a specific gravity of 1.20g/cm³. The purified HBs antigen yielded, in SDS-PAGE (under reducing conditions), four protein bands with apparent molecular weights of 22, 000 and 26, 000 (the two major bands), and 44, 000 and 47, 000. The two proteins of molecular weights of 26, 000 and 47, 000 are likely to be glycosylated, as these were several fold reduced when the cells were cultured in the presence of Tunicamycin. Amino acid analysis, Edman degradation, carboxypeptidase digestion, and ultraviolet absorption spectrum indicated that the HBs antigen from hepatoma cells is very similar to that derived from human plasma.

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Effects of enflurane anesthesia on circulation dynamics and blood catecholamine concentration

Hideki KAJIKI

1989 Volume 101 Issue 7-8 Pages 699-709
Published: August 31, 1989
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.101.7-8_699

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Effects of enflurane anesthesia were studied on the circulation dynamics and blood catecholamine (CA) concentrations during and after inhalation 2% or 3% enflurane in oxygen in 8 dogs at each concentration. During inhalation, the mean arterial pressure (MAP) and cardiac index (CI) decreased significantly. After stopping inhalation, the MAP recovered rapidly to the control values, but the CI recovered to only 65% of the control values. Immediately after starting inhalation, blood CA concentrations decreased significantly in both groups. After stopping inhalation, blood adrenaline (AD) concentration increased rapidly to 6 and 5 times as the control values in the 2% and 3% groups, respectively. Blood noradrenaline (NA) concentration increased to 3 times as the control values in both groups. One hundred and twenty min after starting inhalation, the dogs did not respond to painful stimulation even in the 2% group.
In this study, it has been suggested that the circulation dynamics are suppresed when enflurane has reached more than 0.5 minimum alveolar concentration (MAC) and that the centripetal impulse is thoroughly blocked and CA secretion from the adrenal grand is inhabited at 0.7 MAC of enflurane.

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Left heart bypass with left ventricular failure

A special reference to regional myocardial blood flow

Kazuo SHIOTSU

1989 Volume 101 Issue 7-8 Pages 711-721
Published: August 31, 1989
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.101.7-8_711

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Left heart bypass (LHB) was performed in goats with normal ventricles and in those with LV failure. A pneumatic type pusher-plate pump can be operated in fixed rate, synchronous mode variable pulse rate (VR). In this experiment, the pump was operated in VR, that is, the pusher-plete position signal from a hall effect sensor was utilized to regulate the pump stroke at a constant level, but its rate was allowed to vary automatically depending on the preload and afterload.
LHB was performed in 12 goats with normal ventricles. Blood was drained from the left atrium and was returned to the descending aorta. Pump flows averaged 61.3% of cardiac output (CO). When bypass was started, CO decreased from 72.5 to 57.2 ml/min/kg (p<0.005), and LV^{max dp/dt} decreased from 3943 to 2989 mmHg/sec (p<0.05). Atrial pressure (AP), A-VDO₂, pulmonary atrial pressure (PAP) and TTI did not show significant changes. Regional myocardial blood flow (RMBF), which was measured using the hydogen gas clearance method, decreased from 128 to 85 ml/min/100g (p<0.05).
In 12 goats, LV failure was induced by injecting 0.08 ml/kg of 5N NaOH into the LV free wall. The weight ratio of myocardial necrosis to the LV free wall was 22.7%. Pump flows averaged 52.5% of CO. When bypass was started, CO increased from 45.7 to 61.4 (p<0.005). AP increased from 85.2 to 124 (p<0.001), A-VDO₂ decreased from 6.08 to 4.74 vol% (p<0.01) and PAP decreased from 33.2 to 30.3 (p<0.02). LV^{max dp/dt} and TTI did not show significant changes. RMBF, which was determined at normal LV free wall, decreased from 98.6 to 76.1 (p<0.05). These data suggest that LHB is more effective in goats with LV failure than in those with normal ventricles in terms of left ventricular and systemic circulatory support.

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Studies of iron metabolism of the lung

Part 1. Influence of inhaled iron on iron metabolism of an experimental welder's lung

Kanta FUJIWARA

1989 Volume 101 Issue 7-8 Pages 723-732
Published: August 31, 1989
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.101.7-8_723

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In order to clarify the mechanism of elevated iron stores in welder's lungs, the cellular contents of bronchoalveolar lavage (BAL) fluid, the serum iron, total iron binding capacity, liver non-hemin iron and histological findings were examined in experiments using Wister rats exposed to inhalation of welding fumes for 4, 8, and 12 weeks.
Total cell counts, the percentage of neutrophils and iron laden alveolar macrophages in the BAL fluid increased after continuous inhalation, but they were decreased 3 months after inhalation. Serum iron, transferrin saturation, and liver non-hemin iron in rats exposed to inhalation for 8 and 12 weeks were significantly elevated (p<0.01). Iron stores of rats 3 months after inhalation were elevated more than levels 1 week after inhalation. Histological findings in the lung showed a slight thickening of alveolar wall with cellular infiltration. Iron was deposited predominantly in alveolar macrophages of the alveolar space and in connective tissue macrophages. Iron deposition was also seen in the red pulp of the spleen after inhalation. These data suggest that iron was highly stored in the whole body of patients with welder's lung, and that alveolar macrophages play an important role in the iron metabolism of the lung.

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Studies of iron metabolism of the lung

Part 2. An electron microscopic study of ferritin synthesis in alveolar macrophages

Kanta FUJIWARA

1989 Volume 101 Issue 7-8 Pages 733-742
Published: August 31, 1989
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.101.7-8_733

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In order to clarify the role of alveolar macrophages (alv Mφ) in the iron metabolism of the lung, ultrastructural findings of cells obtained by bronchoalveolar lavage were studied by transmission electron microscopy in rats after inhalation of saccharated iron oxide and welding fumes as well as a patient with welder's lung.
Alv Mφ of rats which had inhaled saccharated iron oxide formed phagosomes and secondary lysosomes 4 hours after inhalation. The first day after inhalation, many ferritin particles were seen in cytosol and formed a diffuse patterns; 19% of alv Mφ had ferritin particles in a paracrystalin arrangement in the secondary lysosomes. Electron micrographs of alv Mφ at 3, 6, 14, and 28 days after inhalation showed an increase of the cells with ferritin particles. Alv Mφ of rats which had inhaled welding fumes phagocytized those iron particles and also synthesized ferritin particles in the cytosol and secondary lysosomes 6 days after inhalation. Numerous ferritin particles were seen in the cytosol and secondary lysosomes of alv Mφ from a patient with welder's lung as well. These results suggest that inhaled iron was phagocytized by alv Mφ and stored in the cytosol by binding with apoferritin which was synthesized by the ribosomes.

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Acute brain swelling and edema by stimulation of the medullary reticular formation in the cold injured brain

Masamitsu KAWACHI

1989 Volume 101 Issue 7-8 Pages 743-762
Published: August 31, 1989
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.101.7-8_743

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The purpose of the present study is to clarify the effects of stimulation of the medullary reticular formation on brain edema and ICP in a cerebral contusion model.
The study was divided into two experiments.
Experiment 1 (32 cats). The effet of stimulation of the medullary reticular formation on tissue water permeability was assessed using normal brain and cold-induced edema. The cold injury was inflicted by applying a freezing probe of -50°C directly on the dura for 1 minute. The animals were divided into 4 groups. Group 1: intermittent electrical stimulation of the reticular formation of the medulla oblongata (P₁₀, L±_2.5, H_-9.0) of a normal brain for 40 minutes; Group 2: cold lesion only; Group 3: same procedure as group 1 on cold-induced edema; Group 4: administration of Angiotensin II for 60 minutes. Water contents of the 4 groups were measured by a gravimetric technique from coronal slices across the lesion.
Experiment 2 (24 cats). The ICP and local cerebral blood volume (CBV) and blood pressure (BP) were continuously measured during stimulation of the medullary reticular formation in animals with cold-induced edema.
The normal water content of the white matter was 67.7±0.9% g water/g tissue. Seventeen hours following cold-induced edema, the water content adjacent to the lesion increased to 71.8% and gradually decreased at areas further from the lesion. There were no significant changes in water content in the contralateral hemisphere. Medullary stimulation of brains with cold-induced edema produced a widespread significant increase in tissue water content in the injured hemisphere by 1.6 to 3.8%. In particular water contents adjacent to the lesion were significantly higher, by 1.3 to 1.8%, than those of group 4 with induced hypertension.
In experiment 2, the BP, ICP and CBV increased abruptly immediately after stimulation and the response patterns were divided into 2 groups. In 15 of 24 animals (62%), temporary elevation of the ICP ranging from 10 to 25mmHg was observed during stimulation with a concomitant increase in CBV and BP. In 9 animals (38%), progressive increase in ICP and CBV were produced by stimulation and even after cessation of stimulation, ICP progressively increased up to 60 to 100mmHg. Progressively increased ICPs were evoked with stimulation of the paramedian reticular formation of the medulla oblongata, that is, the region around the nucleus reticularis parvocellularis.
Stimulation of the medullary reticular formation accelerated brain water permeability and increased CBV, resulting in acute intracranial hypertension in an injured brain.

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Effect of prostaglandin E₁ and prostaglandin F_2α on bone formation of cultured chick embryonic bone

with special reference to the alkaline phosphatase activity

Shigeki UEDA

1989 Volume 101 Issue 7-8 Pages 763-770
Published: August 31, 1989
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.101.7-8_763

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The effects of prostaglandin E₁ (PGE₁) and prostaglandin F_2α (PGF_2α) on bone formation were investigated with femur and tibia from 10-day chick embryo. When the bones were cultured in serum free medium, PGE₁ (10nM) caused a significant increase of bone alkaline phosphatase activity and a slight increase in bone protein content, whereas PGF_2α (100nM) significantly decreased the enzyme activity. On the other hand, in a medium containing 20% serum, PGE₁ did not affect the enzyme activity at 10nM, but slightly increased the activity at 1nM. No effect of PGE₁ on bone calcium content was observed in a concentration ranging from 0.1nM to 100nM. Thus, PGE₁ was likely to stimulate bone formation at 10nM, whereas PGF_2α supressed formation at 100nM.

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Clinical studies of relapse in acute leukemia

Part 1. Cellular composition of bone marrow in remission and prediction of relapse in acute leukemia

Michihiro TAKIZAWA

1989 Volume 101 Issue 7-8 Pages 771-780
Published: August 31, 1989
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.101.7-8_771

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The cellular composition of remission marrow from patients with acute myelocytic leukemia (AML) was studied in order to obtain informations for the prediction of relapse. Hematological findings such as (1) Myeloblast 3%, (2) Promyelocyte 12%, (3) Erythroblasts 45% and (4) Erythroblasts 15% were thought to be atypical in remission marrow. Among these atypical finings, the coexsistence of (1) and (3) or (1) and (4) were important in the prediction of relapse. In the first month after the recognition of (1)+(3) or (1)+(4), the cumulative relapse rate was 35% and 50%, respectively. The rates were 71% and 58% by the second month, respectively. The median remission duration of patients treated within two weeks after the recognition of (1) was longer than that of patients treated after two weeks (4.7 months vs 1.5 months, p<0.05). These results indicate that the coexsistence of atypical findings, (1)+(3) or (1)+(4), are important not only in the predicting of the relapse of AML but also to analyze the dyshemopoiesis of preleukemic states or myelodysplastic syndromes.

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Clinical studies of relapse in acute leukemia

Part 2. Immunosuppressive acidic protein (IAP) in acute leukemia

Michihiro TAKIZAWA

1989 Volume 101 Issue 7-8 Pages 781-789
Published: August 31, 1989
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.101.7-8_781

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Serum levels of immunosuppressive acidic protein (IAP) in 17 patients with acute leukemia were measured. The patients included 5 with acute myelocytic leukemia (AML), 4 with acute promyelocytic leukemia (APL), 3 with acute monocytic/myelomonocytic leukemia (AMoL/AMMoL), and 5 with acute lymphocytic leukemia (ALL). Serum IAP levels in untreated acute leukemia patients were higher than those in normal healthy individuals (763.2±321.1 ng/ml vs 389.8±96.7 ng/ml, p<0.05). They tended to decrease to a normal range in complete remission. A correlation was observed between serum IAP levels and the leukemic cell population in the bone marrow (r=0.39, p<0.05). Serum IAP levels increased in febrile patients compared to those in afebrile patients (p<0.05). Patients with more than 500ng/ml of IAP in remission had more risk of relapse in comparison to those with less than 500ng/ml (7 of 10, 70% vs 2 of 15, 13.3%, p<0.05). These results suggest that the measurement of serum IAP is useful for monitoring the leukemic patient.

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Studies of bronchial asthma

Part 1. Clinical characteristics of late onset asthma

Yukinobu NAKAMURA

1989 Volume 101 Issue 7-8 Pages 791-796
Published: August 31, 1989
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.101.7-8_791

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In order to clarify the immunological characteristics of late onset asthma, 106 cases of asthma in patients over 40 years old were divided into two groups: the early onset asthma group at an age less than 40 years old and the late onset asthma group which was 40 or more years old at onset. The results obtained in each group were compared.
1) Asthma patients with a positive family history or with other allergic diseases in the past history were significantly less in the late onset asthma group compared with the early onset asthma group.
2) An increased eosinophil count in the peripheral blood was observed in both groups with no significant difference.
3) There were more patients with low IgE level (less than 300IU/ml) in the late onset asthma group than in the early onset asthma group.
4) In skin tests to 8 common allergens, the incidence of positive immediate reaction to house dust and aspergillus was low in the late onset asthma group. There was no difference in late or delayed reactions between the two groups.
5) The number of patients with positive serum specific IgE antibody to house dust or mites were few in the late onset asthme group.
6) In the bronchial provocation test with house dust, the proportion of positive response was low in the late onset asthme group.
7) The attack symptoms of asthma were severe and perennial in the late onset asthma group compared with those of the early onset asthma group.

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Studies of bronchial asthma

Part 2. Studies of the changes in the peripheral airway of asthmatic patients by selective alveolo-bronchography (SAB)

Yukinobu NAKAMURA

1989 Volume 101 Issue 7-8 Pages 797-810
Published: August 31, 1989
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.101.7-8_797

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In order to investigate changes in the peripheral airway of asthmatics, selective aveolobronchography (SAB) was performed in 33 cases of asthma. SAB findings were studied in respect to the classification of asthma and also examined with results of pulmonary function test, bronchoalveolar lavage (BAL) and transbronchial lung biopsy.
1. Narrowing of the central airway was observed in almost all cases and 19 cases showed narrowing of the peripheral airways. An uneven appearance of the alveolar figure was observed in 11 cases.
2. Narrowing of the peripheral airway and an uneven appearance of the alveolar figure were often observed in non-atopic patients, in moderate or severe cases and in late-onset asthmatics.
3. The mean value of V₅₀/V₂₅ was increased and the mean value of %V₂₅ was decreased in patients with narrowing of the peripheral airways and also in patients with uneven appearance of the alveolar figure.
4. An increase of neutrophils was noticed in the BAL fluid of patients with narrowing of the peripheral airways.
5. With histological study by TBLB, infiltration of eosinophils into the alveoli as well as mononuclear cells into alveoli and bronchioli was demonstrated in patients with narrowing of the peripheral airways.

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Membrane lipids of Yersinia enterocolitica and influence of culture temperature on membrane lipid composition

Sei-ichiro SHIBUYA

1989 Volume 101 Issue 7-8 Pages 811-820
Published: August 31, 1989
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.101.7-8_811

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The author studied the membrane lipid and fatty acid compositions of Yersinia enterocolitica. Furthermore, the author studied the adaptation of the membrane to changes in culture temperatures, and characterized the physical state of membrane lipid by nylon microcapsule method developed by Okahata and Nagamachi.
Phosphatidylglycerol, phosphatidylethanolamine and cardiolipin were the main phospholipids of the membrane. The ratio of these phospholipid compositions was not changed by culture temperature.
Fatty acids were found to be C_{14: 0}, C_{15: 0}, C_{16: 0}, C_{16: 1}, C_{17: 0}, C_{18: 0}, C_{18: 1} and cyclopropane C_{17: 0}. The last was identified by gaschromatography-massspectrometry and hydrogenation.
In 37°C cultures, saturated fatty acids were increased and unsaturated fatty acids were decreased in comparison with those from 25°C cultures. Cyclopropane C_{17: 0} was also increased in 37°C cultures. These changes appear to be necessary for maintaining membrane stability at high temperatures.
Using NaCI-release from 0.2M NaCI-entrapped nylon microcapsules coated with the membrane lipids as an indicator, it was found that phase transition of lipid membranes from 25°C cultures and 37°C cultures occurred approximately at 30°C and 45°C, respectively. The differences in the phase transition temperature seem to correlate with the adaptive changes in fatty acid composition by culture temperatures.

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Parasitic diseases brought to the Department of Parasitology, in five years, 1984-1989

Hiroyuki MATSUOKA, Toshiki AJI, Akira ISHII, Yasumasa TONGU, Nobuo OHT ...

1989 Volume 101 Issue 7-8 Pages 821-830
Published: August 31, 1989
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.101.7-8_821

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From April 1984 to March 1989, we accepted 399 samples and specimens from hospitals and clinics, and diagnosed 86 cases of parasitic diseases. In this paper, 14 case-reports of zoonoses, larva migrans, imported disease and opportunistic infection are presented and discussed.

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Medical service situation by city and town: A classification attempt using principal component analysis

On out-patient medical treatment in 43 cities and towns of Kagawa Prefecture

Fumihiko JITSUNARI, Fumiyuki ASAKAWA, Taichi NAKAJIMA, Yoshiki MANABE, ...

1989 Volume 101 Issue 7-8 Pages 831-841
Published: August 31, 1989
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.101.7-8_831

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The medical service situation of out-patient medical treatment in 43 cities and towns of Kagawa Prefecture was analyzed, and classified using principal component analysis. When viewed by the average of 43 cities and towns, the rate of dependence on one's own area was 48.4%, the rate of patients from one's own area at 73.3%. Generally, the self-sufficiency of medicalcare was high, and the medical treatment zone was wider in the city region compared with the county region. The medical service situation in 43 cities and towns was classified into 5 types such as the Nuclear type: 4 cities; the Base type: 1 city and 4 towns; the Averageother type: 18 towns; the Peripheral type (the Insular region type: 3 towns; the Peninsular or remote place type: 2 towns; the Satellite city type: 2 towns) and the Depopulated type: 9 towns, using principal component analysis with the rate of dependence on one's own area, rate of patients from one's own area, dependence entropy and medical treatment zone entropy as the input data.

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Variations in blood cholinesterase activity level in termite control workers using chlorpyrifos

Fumihiko JITSUNARI, Fumiyuki ASAKAWA, Taichi NAKAJIMA, Junko SHIMADA, ...

1989 Volume 101 Issue 7-8 Pages 843-852
Published: August 31, 1989
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.101.7-8_843

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In order to clarify the effect of chlorpyrifos on workers using chlorpyrifos for termite control, changes in the cholinesterase activity in the blood of 6 workers was surveyed. Cholinesterase activity in the plasma and erythrocytes decreased in the busy season from May to September and recovered in the off season of January. The ratio of the lowest level of plasma cholinesterase activity from a worker in the busy season to the average activities of a worker examined several times in the off season, was examined in each workers. The ratio was under 50% in 6 workers using chlorpyrifos. The ratio of the mean cholinesterase activity in the erythrocytes of a worker in the busy season to that in the off season was examined, and was about 70% in 6 workers. A negative correlation between working time and increasing ratio of plasma cholinesterase was significant. A marked change in the subjective and objective symptoms of workers was not absent in busy season. Although some workers showed a decrease in the number of erythrocytes and leukocytes, and an increase in serum lipase activities, and abnormalities in the serum lipids. The data indicated that plasma cholinesterase activity levels are useful for biological exposure monitoring of chlorpyrifos.

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Changes in the subjective symptoms, flicker value and the excretions of urinary 17-OHCS, protein and creatinine after physical and mental stress

Seitaro NAKAE

1989 Volume 101 Issue 7-8 Pages 853-862
Published: August 31, 1989
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.101.7-8_853

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Ten students who belong to the volleyball club in a women's university practiced vollyball for 2 hours; 35 female students were walked for 1 hour (Mean heart rate: 105 beats/min). 32 and 6 female students took the one digit addition of Kraepelin's test for 60 and 90 min, respectively. The number of complaints of subjective symptoms, flicker fusion value, the excretion rate of urinary 17-OHCS (17-hydroxycorticosteroids), protein and creatinine before and after physical or mental work were investigated.
The questionnaire included thirty questions, divided into three groups each consisting of ten symptoms, I: drowsiness and dullness, II: difficulty in concentration, III: projection of physical disintegration.
The results obtained were as follows:
1) The number of subjective symptoms in those who walked were particulary increased in groups I and III. Group II symptoms were increased in the participants who calculated the one digit addition.
2) The flicker fusion value after physical exercise was not decresed, but in one digit addition the flicker fusion value was significant decreased after working.
3) The excretion of urinary 17-OHCS tended to decrease after vollyball training and walking. In the case of one digit addition, the excretion of urinary 17-OHCS was increased.
4) The excretion of urinary protein was generally increased after vollyball training, and urinary creatinine was increased after physical and mental work.
In conclusion a difference was demonstrated between physical and mental work and related complaints of subjective symptoms, flicker fusion values and excretion rates of urinary 17-OHCS.

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