Inactive form of erythrocyte carbonic anhydrase B in patients with primary renal tubular acidosis.

Evidence was found for an inactive form of carbonic anhydrase type B in the erythrocytes of two children with primary renal tubular acidosis. The addition of zinc chloride to hemolysates from these patients resulted in a marked increase in the activity of this enzyme. No such effect was noted with hemolysates of control subjects. No significant differences were observed in the zinc levels of hemolysates of these patients and of normal individuals. However, the level of zinc in the carbonic anhydrase B isolated from one of these patients was low, suggesting a modified form of the enzyme. The restoration of activity upon the addition of zinc was reversed by ethylenediamine tetraacetate, but no such effects were noted for the carbonic anhydrase B of normal individuals. Thus the abnormal carbonic anhydrase B has decreased zinc binding. The ultraviolet difference spectrum of the carbonic anhydrase B of normal individuals and that of a patient showed a peak at 305 nm which decreased upon the addition of zinc. The abnormal form of carbonic anhydrase B was not distinguishable from that of normal individuals by either immunological or electrophoretic criteria.

Risk factors for alloimmunization by patients with sickle cell disease.

Blood transfusion in patients with sickle cell disease (SCD) is limited by the development of alloantibodies to erythrocytes. In the present study, the frequency and risk factors for alloimmunization were determined. Transfusion records and medical charts of 828 SCD patients who had been transfused and followed at the Belo Horizonte Blood Center, Belo Horizonte, MG, Brazil, were retrospectively reviewed. Alloimmunization frequency was 9.9% (95% CI: 7.9 to 11.9%) and 125 alloantibodies were detected, 79% of which belonged to the Rhesus and Kell systems. Female patients developed alloimmunization more frequently (P = 0.03). The median age of the alloimmunized group was 23.3 years, compared to 14.6 years for the non-alloimmunized group (P < 0.0001). Multivariate analyses were applied to the data for 608 hemoglobin (Hb) SS or SC patients whose number of transfusions was recorded accurately. Number of transfusions (P = 0.00006), older age (P = 0.056) and Hb SC (P = 0.02) showed independent statistical associations with alloimmunization. Hb SC patients older than 14 years faced a 2.8-fold higher (95% CI: 1.3 to 6.0) risk of alloimmunization than Hb SS patients. Female Hb SC patients had the highest risk of developing alloantibodies. In patients younger than 14 years, only the number of transfusions was significant. We conclude that an increased risk of alloimmunization was associated with older patients with Hb SC, specially females, even after adjustments were made for the number of transfusions received, the most significant variable.

Studies on the tissue thromboplastin during the coagulation-fibrinolytic process--ultrastructural changes.

An electron-microscopical study was made on morphological changes in which T-Tbp would undergo during clotting and fibrinolytic process. Morphological appearance of concentrically arranged membrane structure in T-Tbp remained nearly intact during blood coagulation process. T-Tbp, which existed in the sediments following dissolution of fibrin clot by the application of UK, showed an appearance of fine granules adhering to the surface of aggregates of particles through SEM. Through TEM, T-Tbp in the sediments was found to have retained its concentrically arranged membrane structures in most places, while, in some other places the appearance of fused membranes, smaller single vesicles and long sheets of membranes, and the formation of "blebs" etc. were observed. Various morphological changes caused by fibrinolytic substances accompanied the loss in coagulation activities. Our results showed that coagulation activities of T-Tbp must be completely dependent upon the presence of the membrane structures.

Ultrastructural changes of the tissue thromboplastin after intravenous injection in rabbits.

Attempts were made to demonstrate ultrastructural changes of the tissue thromboplastin after intravenous injection, as a model experiment on the pulmonary microthrombi formation induced by the tissue thromboplastin circulating from venous return. Concentrically arranged membrane structures of the injected thromboplastin disappeared in extremely short time after the injection of the thromboplastin in rabbits. The long sheet membrane of the injected thromboplastin was frequently seen as adhered to the vascular endothelium or to the surface of blood corpuscles. Furthermore, fibrin fibres were formed in contact with the long sheet membrane of the thromboplastin. Membrane structures were not found anywhere in the control rabbits.

Analysis of the disappearance curve of labelled fibrinogen at the time of hyperfibrinogenemia in rabbits with acute or chronic intravascular coagulation.

In order to study fibrinogen metabolism, the disappearance curve of 125I-abelled homologous fibrinogen was investigated in the rabbits with experimentally induced acute or chronic intravascular coagulation by injection of Lycopodium spores or thromboplastin. The results obtained were as follows. 1. Using haemolysate, an intermediate phase with upward convexity was clearly recognized between the early rapid-decay phase and the late slow phase in each radioactivity decay curve obtained in groups of rabbits. This convexity was most marked with acute intravascular coagulation induced by injection of Lycopodium spores, and was less marked, although higher, with chronic intravascular coagulation induced by injection of thromboplastin than that in the normal control. 2. The disappearance curve with the intermediate phase could be expressed, in approximation, as a sum of 2 equations--the initial exponential decay equation and the late parabolic one. 3. From the results obtained by separate examinations of the disappearance curve of plasma, fibrin clots and serum in rabbits with acute intravascular coagulation induced by injection of Lycopodium spores, the intermediate phase appears to be influenced more by the secondary increase of labelled non-clottable part tahn recirculation of the labelled fibrinogen. 4. The half time (27.4 hours) of the radioactivity in fibrin clots at the late phase observed in the group of rabbits with acute intravascular coagulation induced by injection of Lycopodium spores was shorter than that in the normal rabbits (50.2 hours). This fact may indicate that the increase of fibrinogen in the group of rabbits with acute intravascular coagulation induced by the injection of Lycopodium spores is due to overproduction of fibrinogen. 5. The half time of labelled fibrinogen should be calculated from disappearance curve of fibrin clots, instead from that of haemolysate or plasma.

Summer-type hypersensitivity pneumonitis. A unique disease in Japan.

A unique form of hypersensitivity pneumonitis in which clinical symptoms appear in the summer and subside spontaneously in the mid-autumn was found in Japan. This disease was named summer-type hypersensitivity pneumonitis and was found the most prevalent form in Japan. This disease has the following characteristic features: 1) initiation in the summer; 2) repeated episodes during subsequent seasons for many years; 3) familial occurrence; 4) no occupational relationship; 5) positive returning-home provocation test; 6) cough, dyspnea and remittent fever as a clinical triad; 7) diffuse nodular shadows on chest x-ray film; 8) leukocytosis with neutrophilia; 9) moderately decreased % VC and markedly decreased Dco and PaO2; 10) skin reactivity to PPD is negative while symptomatic; 11) pulmonary lesions of biopsied specimens show epithelioid cell granulomas without central necrosis (63.3 percent), plus alveolitis and/or pneumonitis; 12) isolation of patients from their home environment diminishes symptoms; 13) corticosteroid is effective; 14) seasonal atmospheric microbiological pollution is speculated upon, but the offending antigen is not defined yet.

Exercise and oxygen inhalation in relation to prognosis of chronic obstructive pulmonary disease.

We examined the relationship between prognosis and arterial blood gases during exercise and 100 percent oxygen inhalation in 54 patients randomly selected from 119 background patients with chronic obstructive pulmonary disease (COPD). Light exercise was performed and 100 percent oxygen was inhaled during clinically stable stages. By four years after these tests, 19/54 patients had died from respiratory failure. All subjects had similar physical and clinical features. Survivors had significantly higher PaO2 during air breathing than nonsurvivors; in nonsurvivors, FEV1 and MVV were significantly lower, and heart rate and RV/TLC were significantly higher. Exercise PaO2 of nonsurvivors decreased by 6.7 mm Hg, whereas that of survivors did not change. The P(A--a)O2 did not change in survivors and nonsurvivors during exercise, but in survivors it was significantly smaller. Mean PaO2 after 100 percent oxygen was significantly lower, and PaCO2 was significantly higher in nonsurvivors than in survivors, and in nonsurvivors the increase in PaO2 during 100 percent oxygen correlated positively with the time between first admission and death. These results indicate that patients with combination of resting arterial hypoxemia, worsened hypoxemia during stepped-up exercise, and lesser degree of arterial oxygenation and increased PaCO2 during 100 percent oxygen inhalation may have a poor prognosis.

Effect of thyroid hormone on the levels of erythrocyte carbonic anhydrase isozymes and 2,3-diphosphoglycerate in rabbits.

Levels of rabbit erythrocyte carbonic anhydrase B and C isozymes were determined in experimental hyperthyroidism using a quantitative immunologic technique. Levels of erythrocyte 2,3-diphosphoglycerate and protein binding iodine were simultaneously determined. Thyroxine and 3,5,3'-triiodothyronine were administered to rabbits orally for 30 days. A significant decrease in carbonic anhydrase B type was observed after 30 days, although no significant change was observed in carbonic anhydrase C type. These findings suggest that the steady state level of carbonic anhydrase B type in red cells is affected by thyroid hormone more readily than that of carbonic anhydrase C type. The level of red cell 2,3-diphosphoglycerate increased markedly after 10 days of treatment, corresponding to the increase of protein binding iodine. The clinical or pathologic significances were discussed in relation to the changes in the levels of these isozymes and 2,3-diphosphglycerate in red cells.

Effects of long-term administration and withdrawal of clonidine on activity of sympathetic efferent nerve unit in spontaneously hypertensive rats.

Daily administration of clonidine for 5 weeks (approx. 300 micrograms/kg/day, p.o.) produced significant decreases in blood pressure and renal sympathetic efferent activity in spontaneously hypertensive rats (SHR). After cessation of clonidine administration, blood pressure and sympathetic nerve activity recovered rapidly from the depressed levels to the untreated control levels. These findings could indicate that clonidine-induced hypotension and its rapid recovery to control after withdrawal of clonidine are caused by corresponding changes in sympathetic efferent nerve activity.

Estimations of active and inactive carbonic anhydrase isozyme B in human red cells.

Human erythrocyte carbonic anhydrase isozyme B was measured by a specific and immunological method. The levels of carbonic anhydrase B were determined in normal subjects, patients with hyperthyroidism, and patients with chronic obstructive lung disease and patients with epilepsy under treatment with acetazolamide, using the rapid assay method of single radial immunodiffusion. The levels of carbonic anhydrase B showed a decrease in hyperthyroidism, and increased in chronic obstructive lung diseases and epilepsies. Closely negative correlations were observed between carbonic anhydrase B and T3 resin sponge uptake or protein bound iodine (PBI) levels in hyperthyroidism. Simultaneously, carbonic anhydrase B-dependent esterase activity (active carbonic anhydrase B enzyme) was determined kinetically using the immunoadsorbent method in the above-mentioned patients. The results were compared with the total enzyme protein (active and inactive carbonic anhydrase B enzyme), estimated by the single radial immunodiffusion technique. In patients treated with acetazolamide, the "true" specific activity of the carbonic anhydrase B (carbonic anhydrase B-dependent esterase activity/total carbonic anhydrase B protein) decreased remarkably. In chronic obstructive lung disease, no remarkable changes were observed. On the other hand the specific activity was elevated in hyperthyroidism to 2 times higher than that of normal subjects. The clinical significance of these active enzymes is discussed.

Evaluation of volatile sulfur compounds in the expired alveolar gas in patients with liver cirrhosis.

Trace amounts of volatile sulfur compounds were determined in the expired alveolar gas by gas chromatography. Among these sulfur containing substances, methyl mercaptan and dimethyl sulfide were quantitatively analyzed in 116 subjects; 53 normal, 13 acute hepatitis, 11 chronic hepatitis, 20 hepatic cirrhosis, and 19 stomach ulcer and/or biopsy of gastric mucosa. Fasting level of dimethyl sulfide in cirrhotics was 4.05 +/- 1.06 ng/dl, significantly elevated by comparison with normal controls (1.54 +/- 0.09 ng/dl) (P less than 0.05). In contrast, methyl mercaptan did not show a statistically significant rise in this study. The implications of the significant increase in dimethyl sulfide concentration in liver cirrhosis are discussed.

Clinical application of breath analysis for dimethyl sulfide following ingestion of DL-methionine.

After overnight-fasting, the concentration of dimethyl sulfide in expired alveolar gas (alv-DMS) was determined serially following ingestion of 2 g of DL-methionine in normal subjects and patients with liver diseases. Alv-DMS rose to a peak in 30 to 90 min, declined markedly within 3 h, and then decreased gradually. Half-disappearance times (T 1/2) (mean +/- S.E. min) in each experimental group were: normal (N = 9) 61.7 +/- 4.7, acute hepatitis (N = 6) 62.5 +/- 6.8, chronic hepatitis (N = 10) 84.0 +/- 13.0, and liver cirrhosis (N = 13) 159.2 +/- 30.4, respectively. Cirrhotics had a T 1/2 significantly longer than that of the other three groups: vs. normal P less than 0.02, vs. acute hepatitis P less than 0.05, and vs. chronic hepatitis P less than 0.05. T 1/2 of alv-DMS following ingestion of DL-methionine seems to be of clinical interest.

Effect of single oral administration of nilvadipine on cerebral blood flow in chronic cerebral infarction.

UNLABELLED: The effect of nilvadipine, a newly developed calcium antagonist, on regional cerebral blood flow (rCBF) was investigated in 7 patients with chronic cerebral infarction. rCBF was measured by the 133Xenon inhalation method. Patients were given a single dose of 4 mg of nilvadipine after the first measurement of rCBF, and the second measurement was done one hour after the administration. All patients had hemiparesis and 2 of them had mild to moderate mental deterioration, but all patients could walk to the outpatient clinic by themselves. RESULTS: (1) rCBF of the affected side significantly increased by 22.7% after single oral administration of nilvadipine (p < 0.05). The increase of rCBF was significantly marked in frontal regions of the affected hemispheres. (2) No significant changes in blood pressure or end tidal partial pressure of carbon dioxide were observed during the examination. These results indicate that nilvadipine has a potent selective vasodilatory action on the cerebral arteries in patients with cerebral infarction.

The effect of enalapril maleate on cerebral blood flow in chronic cerebral infarction.

UNLABELLED: The influence of long-term oral administration of enalapril maleate (an angiotensin II-converting enzyme inhibitor) on regional cerebral blood flow (rCBF) was studied in 10 patients with chronic cerebral infarction. The rCBF was measured by a 133Xe inhalation method before and after a mean of sixty-five days' administration of 5 mg of enalapril. RESULTS: Mean arterial blood pressure (MABP) was mildly decreased in 6 patients, but the average change in MABP was not significant (Endtidal partial pressure of carbon dioxide (PeC02) was not changed significantly. The mean rCBF was increased by 8% after administration of enalapril (0.05 less than p less than 0.1) There was no significant correlation between percent change in MABP and the percent change in rCBF. These results indicate that enalapril has not only antihypertensive action but also a beneficial effect on the cerebral circulation in patients with chronic cerebral infarction.

Relationship between cytomorphologic features and prognosis in small-cell carcinoma of the lung.

Small-cell bronchial carcinoma cells obtained from 18 patients by fiberbronchoscopic brushing were examined to determine whether the cytomorphologic features could be related to the prognosis. Nuclear features such as small (less than 10 micrometer) or large (greater than 11 micrometer) size, round or aberrant shape and dark or light staining, were evaluated. In small-cell carcinoma cases with a very poor prognosis (death within four months), the incidence of small, round, darkly stained nuclei was much greater and a coarse or pyknotic chromatin pattern was more frequent in comparison with small-cell carcinoma cases with a better prognosis (survival greater than two years). In cases with a good prognosis, the incidence of large, lightly stained nuclei with aberrant shapes was much greater and a fine chromatin pattern was more frequent in comparison with cases with a very poor prognosis. Pertinent cytomorphologic features thus seem to be related to the prognosis.

New wound-healing model using cultured corneal endothelial cells. 1. Quantitative study of healing process.

A new in vitro model to evaluate the corneal endothelial wound-healing process was developed. Confluent monolayer cell sheets of either rabbit or bovine corneal endothelial cells were cultured on cover slips, the surface of which had been precoated with hydroxyethyl methacrylate (HEMA) except for the 8 mm-diameter area for cell culture. Wounds were made by scraping off cells in the center of the cell sheets using a rotating silicone tip. The areas of these wounds were measured by a method of computed planimetry. The average wound area was 3.00 +/- 0.21 mm2 in rabbit cell cultures and 3.18 +/- 0.28 mm2 in bovine. The healing process after the wounding was observed using an inverted phase contrast microscope and wound-healing rates (healed area per hour) were calculated. Wounded areas were recovered by 60 hours in rabbit experiments, and by 84 hours in bovine experiments. Wound-healing rates were different among the observation periods and showed differences between the two species. The healing rates in the rabbit were higher than those in the bovine in the early phase. This in vitro model can be used for quantitative evaluation of the wound-healing process of the corneal endothelium.

New wound-healing model using cultured bovine corneal endothelial cells. 2. Role of migration and mitosis studied by immunohistochemistry.

The roles of migration and mitosis in the in vitro corneal endothelial wound-healing process were studied using an immunohistochemical method. The in vitro wound-healing model was a cultured cell sheet prepared using bovine corneal endothelium, and the wound was made at the center of the cell sheet using a rotating silicon tip. Following the wounding, specimens were incubated with bromodeoxyuridine (BrdU) for 12 hours. After the fixation with 10% phosphate-buffered formalin, the cells incorporating BrdU were stained by the avidin biotin peroxidase complex method. In 12 hours after the wounding, no stained cells were observed. In 24 hours, cell migration had started and a few cells were stained; the average number of stained cells was 14.6 +/- 4.0 cells/mm2. In 48 hours, the number of stained cells increased. The maximum number of stained cells was observed between 48 and 60 hours after the wounding. The number of stained cells decreased rapidly after 72 hours when the wound was almost closed. Stained cells were most numerous in the area within 0.8 mm of the wound edge throughout the wound-healing. The endothelial wound-healing process in this model could be divided into four phases, latent, migration, concurrence of migration and mitosis, and inhibition phases. This in vitro wound-healing model could simulate the wound-healing process in vivo and will be useful for a quantitative evaluation of drug effects on the corneal endothelial wound-healing.

Corneal endothelial cell damage in penetrating keratoplasty.

Fifty-eight corneal grafts were examined by specular microscopy to determine the corneal endothelial cell damage following penetrating keratoplasty. The average postoperative follow-up period was 10.1 months. The cell density decreased continuously during the observation period, and the endothelial cell loss of the central area of the cornea in all the cases averaged 10.4% at two weeks after the surgery, 16.0% at one month, 33.0% at three months, 39.4% at six months, and 48.2% at one year. However, the cell loss in the keratoconus group averaged -1.9, 1.2, 9.9, 30.6 and 33.4% for the corresponding postoperative periods. The cell loss in the bullous keratopathy eyes showed higher values compared to those in the keratoconus and the corneal leukoma eyes. It was concluded that cell loss in penetrating keratoplasty during the first postoperative year depends on the host diseases.

A cross-cultural comparison of British and Japanese lay theories of schizophrenia.

Nearly 300 matched British and Japanese participants completed an equivalent three-part questionnaire in their native language. The questionnaire covered general beliefs or conceptions about schizophrenia, causal explanations for the aetiology of schizophrenia, as well as the role of hospitals in particular, and society in general, in helping schizophrenics recover from their illness. It was predicted that the Japanese, who have more taboos about mental illness than the British, would see schizophrenics as more difficult, dangerous and "morally insane". Factor analysis of each of the three parts of the questionnaire yielded a clearly interpretable structure. The British were more concerned with the rights of schizophrenics and believed them to be less dangerous and abnormal than did the Japanese. The Japanese favoured sociological (stress) explanations more than the British for the cause of schizophrenia. Whereas the Japanese saw micro- and macro-society change as the best way to help schizophrenics, the British stressed individual care and consideration as more relevant.

[Food-borne outbreaks of gastroenteritis caused by small round structured viruses. 1. Four outbreaks of gastroenteritis associated with oyster consumption].

Between December 9, 1988 and January 28, 1989, there were four outbreaks of acute gastroenteritis in Saitama prefecture. Eighty-two of 123 persons (67%) attending four banquets in restaurants became ill: 44 cases attending three banquets were related to eating raw oysters, and 38 attending one banquet to eating sashimi. The most common symptoms were nausea, diarrhea, abdominal cramps, and vomiting. Average incubation periods were 29 to 32 hours long. Bacteriologic analysis of stool specimens did not reveal causative agents. Small round structured viruses were detected in fecal specimens from 19 of 39 ill persons (49%) by electron microscopy. In one of four outbreaks, the formation of antibody to small round structured virus in paired serum samples was detected by western blot test. Small round structured viruses were implicated as the etiologic agents in four outbreaks of acute gastroenteritis.