The effect of electrical stimulation of the internal capsule (IC) or sensory thalamic nucleus (VPM) on neuronal activity in the subnucleus caudalis of the spinal trigeminal nucleus (STNcd) was examined electrophysiologically in the cat. The nociceptive neuronal firing in the STNcd was suppressed by the facilitation of the non-opiate pain inhibitory system, such as electrical stimulation of the IC or VPM of either side. High amplitude spontaneous and continous neuronal hyperactivity (deafferentation hyperactivity: DH), which was provoked in the left STNcd after complete ablation of the left Gasserian ganglion, was conspicuously suppressed by electrical stimulation of the IC of either side (rt. IC stimulation: 30/60; It. IC stimulation: 13/33), as well as by VPM stimulation of either side (rt. VPM stimulation: 5/13; It. VPM stimulation: 5/9). DH, similar to that provoked in the left STNcd after ganglionectomy, was found to spread to the right sensory thalamic nucleus. Twelve neurons with DH were identified in right VPM and nucleus centralis medialis. As many as nine of these twelve neurons were distributed in the marginal area of the VPM. DH in the right thalamus was also suppressed by the stimulation of the IC on either side, however, the suppressive effect was predominant with ipsilateral IC stimulation. These results are compatible with the clinical experience of relief of deafferented (deafferentation) pain by electrical stimulation of the IC or sensory thalamic nucleus (VPM, VPL). After detailed analysis and considerations, it was supposed that this experiment supports the postulated theory that not only deafferented pain but also afferent pain is relieved by the facilitation of the non-opiate pain inhibitory system including the IC and/or the sensory thalamic nucleus.
The purpose of this study is to disclose the differences of risk factors between pre-term low birth weight (LBW) infants and full-term LBW infants. The cases studied were all singleton, live, LBW infants born in 2 districts of a certain city from 1986 to 1987. Each case was matched with 2 controls by district, sex and birthday, and a questionnaire concerning the factors related to birth weight was sent to both cases and controls. The questionnaires were answered by 308 (76.6%) cases and 644 (80.1%) controls. The cases and controls who responded were matched again, and the case group was divided into a pre-term group and a full-term group. And each case group was compared with the same control group. The results were as follows. 1) Factors related to a higher incidence of pre-term LBW infants were different from those of full-term LBW infants. 2) The risk factors for pre-term LBW infants were male sex; increased parental age, parental short height and low academic career; maternal smoking, anxiety and night working during pregnancy; and toxemia. 3) The risk factors for full-term LBW infants were female sex; parental short height, light weight and smoking; maternal young age and work during pregnancy; toxemia and hypertension. 4) Discriminant analysis disclosed that toxemia, paternal short height and maternal night work during pregnancy were strongly related to a higher incidence of pre-term LBW infants, and that maternal and paternal light weight, maternal smoking were the more important factors related to a higher incidence of full-term LBW infants.
To evaluate IgG-related mechanisms in bronchial asthma, Candida-specific IgG antibodies were estimated in 130 patients with bronchial asthma, using an enzyme-linked immunosorbent assay. In 41 healthy subjects, the levels of Candida-specific IgG antibodies were 0.595±0.465. In 130 asthmatics, the levels of Candida-specific IgG antibodies were 0.792±0.524. The difference in the levels of Candida-specific IgG antibodies between healthy subjects and asthmatics was more remarkable in cases from 0-20 years and 41-50 years of age. In cases with a negative delayed skin reaction to Candida, the levels of Candida-specific IgG antibodies were higher than in cases with a positive delayed skin reaction. There was a slight, but not significant correlation between the levels of Candida-specific IgG antibodies and disease severity. There was a significant correlation between the levels of Candida-specific IgG antibodies and precipitating antibodies to Candida. In conclusion, the analysis of Candida-specific IgG antibodies is useful in the evaluation of the pathophysiological mechanisms of bronchial asthma.
To clarify the characteristics of Candida (C.) albicans as an allergen in bronchial asthma, C. albicans-induced histamine release from basophils was observed in 35 healthy subjects and 60 asthmatics. In 35 healthy subjects, the mean %maximum histamine release was 2.5±2.9, and there was no significant histamine release (more than 15%) after C. albicans stimulation. Sixteen of 60 asthmatic cases showed a significant histamine release induced by C. albicans. The frequency of cases with significant histamine release was the highest in patients between 41 and 50 years old. Histamine was released more generally in cases over 41 years. Histamine was released not only in early onset cases with higher levels of serum IgE, but also in late onset cases with lower levels of serum IgE. There was a significant correlation between the level of C. albicans-specific IgE antibodies and histamine release. However basophils in some cases with negative specific IgE antibodies released a significant amount of histamine. In conclusion, histamine release induced by C. albicans is useful in the evaluation of the pathophysiological mechanisms of bronchial asthma.
The effects of pretreatment with either SCH 23390, a selective D-1 antagonist or YM-09151-2, a selective D-2 antagonist, on methamphetamine (MAP)-induced changes in the substance P and TRH system were investigated. A single dose of 4 mg/kg of MAP reduced the striatal level of substance P 30 min after injection. This reduction was reversed by pretreatment with YM-09151-2 but not with SCH 23390. Repeated administration of 4 mg/kg of MAP for 14 days reduced specific substance P and TRH binding in the striatum at 1 h, but not at 48 h, after the last injection. The reduction of striatal substance P receptor binding was prevented by pretreatment YM-09151-2 but not SCH 23390 prior to each MAP administration. The reduction of striatal TRH receptor binding was prevented by pretreatment with either antagonist. These results indicate that changes in striatal substance P level and substance P receptor binding induced by subchronic MAP treatment are mediated via activation of D-2 but not D-1 receptors by MAP-released dopamine, while changes in striatal TRH receptor binding are mediated via both D-1 and D-2 receptors.
A cell line, HuH-33, was established in vitro from a patient with hepatocellular carcinoma (HCC), who had been treated with some chemotherapeutic agents. The chromosome number was widely distributed in 6888. This cell line has grown slowly, exhibiting a doubling time of approximately 150h, was intransplantable into nude mice, and secreted alpha-fetoprotein, albumin, β2-microglobulin, ferritin, tissue peptide antigen and extracellular matrix materials. HuH-33 was polygonal or spindle shaped, but it was keratin-positive and desmosome-like structures were observed with transmission electron microscopy, suggesting an epithelial origin of HuH-33.
In an attempt to evaluate the effectiveness of mass screening for lung cancer, survival was compared between 276 patients with lung cancer detected by mass screening (mass screening group) and 265 patients detected by cancer-related symptoms (symptomatic group) from 1976 to 1984. In the mass screening group 61% of the patients were Stage I, while in the symptomatic group only 27% were Stage I (p<0.01). The resectability and curative resection rate in the mass screening group were 63% and 43%, respectively, significantly higher than the 38% and 21% rates in the symptomatic group (p<0.01). The 5-year survival rate in the mass screening group was 34%, compared to 17% in the symptomatic group (p<0.001). These results indicate that mass screening for lung cancer results in earlier detection and improved prognosis.
In order to improve the efficiency and quality of mass screening for lung cancer and to establish a new system called the “Okayama Lung Project”, examinations which included dual and comparative reading of annual chest X-ray films, direct patient contact, and sputum cytology were studied. Mass screening for lung cancer was conducted utilizing annual chest X-ray films obtained during mass screening for tuberculosis. A series of films was read and the identification numbers of each film showing evidence of any disease were recorded. Later, the entire series were independently read by the another radiologist blinded with respect to the first diagnosis (dual reading). Any chest X-ray with abnormal findings detected by dual reading was then compared with a previous film (comparative reading). For patients suspected of having lung cancer, direct contact was performed by the institution which would make a definitive diagnosis. Sputum cytology examination was conducted in the group at high risk for the hilar lung cancer. Using this new system, the time from mass screening to surgical resection was shortened by 50 days, and the detection rate in the high risk group for hilar lung cancer was elevated from 37.77/100, 000 to 46.44/100, 000. These results indicate that the Okayama Lung project is a satisfactory standard mass screening system for lung cancer.
The three-dimensional architecture of the collagenous framework of triangular fibrocartilage (TFC) of the human wrist joint was examined using polarized light microscopy and scanning electron microscopy (SEM). Polarized light microscopy of sagittal sections demonstrated collagen fibers consisting of many layers, while observations of horizontally sectioned samples demonstrated collagenous fibrils arranged in concentric circles. The circular center was slightly deviated to the radial side, while a coronal section demonstrated few collagenous fibrils running in that direction. Scanning electron micrographs of the carpal surface revealed a network of collagenous fibrils with collagenous fibrils oriented in one direction under this mesh. In the ulnar surface, the collagenous bundles made of over 100 fibrils formed a textile structure. The cut surface was made of collagenous fibrils running in one direction. Degenerative changes are more frequent and more intense on the ulnar surface, and the slits are frequently situated in the radial side of the TFC. The results of the fibrillar framework were discussed in relation to mechanical functions and injuries of the TFC in the wrist joint.
The study evaluated the impact of the October, 1984 introduction of a 10% cost sharing on the attitude of insured persons in the Employees' Health Insurance Scheme. The subjects were 1701 Health Insurance Societies, all of which had joined the Health Insurance Society Association from 1983 to 1985. Physician utilization rates in 1983 and 1984 were analyzed to disclose the effects of the 10% cost sharing introduction. Moreover the author examined which factors of Health Insurance Society were related to the changes of the rates by multiple regression analysis. The results are as follows. 1. All medical service rates of admissions, outpatient visits, and dental visits were reduced significantly after the introduction. 2. The impacts were higher in the Societies in which rates before the introduction were higher for all medical services. 3. Multiple regression analysis revealed differences in indicators related to the characteristics of the society among all medical service rates.
An immunohistochemical study of transthyretin (TTR), synthesized in the choroid plexus epithelia, was performed in normal brain tissue and 92 brain tumors. Glial fibrillary acidic protein (GFAP) and cytokeratin (CKER) were also examined in 11 choroid plexus papillomas (CPPs) and 19 ependymomas. A positive TTR reaction was observed diffusely in the majority of normal choroid plexus epithelia. All of the 11 cases of CPP were positive for TTR, but the other tissues and tumors (including ependymomas) were negative for TTR. GFAP -positive cells were occasionally found in five of the CPPs. The majority of the GFAP -positive cells were also positive for CKER, but not for TTR. There was an inverse relationship between the intensity of immunostaining for GFAP and for TTR in CPPs. Among the 19 cases of ependymomas, 16 were positive for GFAP, 3 for CKER, but none for TTR. TTR can be a very useful and diagnostic marker of CPP. A consective semithin-thin section microscopic method demonstrated abundant secretory granules and well-developed Golgi apparatus in TTR-positive cells. By immunoelectron microscopy using a pre-embedding method, a positive reaction for TTR was observed in the secretory granules. This result suggested that CPP cells as well as choroid plexus cells synthesize TTR de novo.
The optimal fat/glucose ratio for TPN was determined, using oxidation and distribution of 14C labeled fat emulsion and glucose. Animals were divided into five groups according to percentage of energy supplied as fat i·e. 0%, 20%, 40%, 60% and 80%, respectively. 14C-labeled fat emulsion and glucose were injected during TPN and the rate of oxidation to 14CO2 as well as 14C distribution in various organs were measured. Oxidation of 14C-glucose was constant in all groups and that of 14C-labeled fat emulsion was inversely related to the amount of glucose administered simultaneously. 14C-distribution in adipose tissue was correlated to the amount of glucose infused. As oxidation of glucose was significantly higher than that of fat, the higher the fat content of non-protein calorie, the less oxidation was observed. The groups given 60% and 80% energy composed of fat, showed less nitrogen retention and glycogen content in liver and muscle. These two groups exhibited high blood levels of free fatty acid, cholesterol and phospholipids. These findings suggest that the optimal blending ratio of fat would be around 20-40% of the non-protein calories.
Adriamycin (ADR)-resistant Ehrlich ascites tumor cells (EATC) were established by repeated exposure to ADR. Using these resistant cells, the modification of the killing effect of ADR by hyperthermia and cepharanthine (CEP) was evaluated. The killing effect of ADR depended on the intracellular uptake and retention of ADR. ADR-resistant EATC had an enhanced acquired capacity that increased the efflux of intracellular ADR. Therefore, the intracellular uptake and retention of ADR were decreased. CEP inhibited the efflux of intracellular ADR and increased the intracellular retention of ADR, thereby enhancing the killing effect of ADR. Hyperthermic treatment increased the influx of ADR, but the killing effect of ADR was not enhanced by hyperthermia alone because of the increased efflux of intracellular ADR. However, with the combination of hyperthermia and CEP, hyperthermia accelerated the influx of ADR, and CEP suppressed the efflux of intracellular ADR. Then the intracellular uptake and retention of ADR were further increased, and the killing effect of ADR was markedly enhanced. Thus the combined use of hyperthermia and CEP reduced the resistance to ADR. These findings suggest that ADR resistance in cancer cells can be overcome by the combination of ADR, hyperthermia and CEP.
Between 1975 and 1986, cementless total knee replacement (Okayama University Mark-II or KC-1) was performed on 38 osteoarthritic knees of 30 patients over 65 years old. Five patients died of unrelated causes from 3 to 6 years after the surgery. A follow up study was carried out for over 3 years in 27 knees of 20 patients. The mean length of follow-up was 69 months, ranging from 36 to 116 months. The average clinical knee score by three university knee score system was 37.1 points preoperatively and improved up to 77.6 points at the follow up. Pain relief was clinically excellent and correction of the deformity was also sufficient. Clinical and radiographic evaluations were also made in correlation with osteoporosis of the knee. According to Foltin's classification of osteoporosis of the knee, operated knees were grouped into five grades (IV), and about 80 percent of the patients had osteoporosis of more than grade III. Radiographically, loosening or migration was observed in 5 knees (19%) in this series, and all of the patients had severe osteoporosis. In other patients with osteoporosis, the clinical knee score improved similarly, but in about half of them there was some change in the femorotibial angle or medial or posterior angle of the tibial component. Therefore, such changes should be followed in radiological examinations in relation to the clinical course.
In this study, patients with diffuse panbronchiolitis (DPB) and bronchiolar type of HTLV-I associated bronchiolo-alveolar disorder (HABA-B) were evaluated in regard to regulation of interleukin-2 receptor (IL-2R) and lymphocyte subsets. The levels of soluble IL-2R in sera and lymphocyte culture supernatant, the responsiveness of lymphocytes to IL-2 and the percentage of IL-2R positive lymphocytes and helper-T lymphocytes were higher in DPB than in normal controls. Furthermore in HABA-B, the levels of IL-2R were much higher than those of DPB. The correlation coefficient between the percentage of IL-2R positive lymphocytes or helper-T lymphocytes and the CD4/CD8 ratio were statistically significant. In the patients with DPB, the percentage of IL-2R positive lymphocytes and helper-T lymphocytes were increased with the progress in clinical stage of DPB or the serum titer of cold hemagglutinin increase. Therefore, the cellular immunity regulated by the IL-2 and IL-2R system might play an important role in the pathogenesis of DPB and HABA-B.
In this study, the Interleukin-2 receptor (IL-2R) and lymphocyte subsets in the patients with idiopathic interstitial pneumonia (IIP), interstitial pneumonia with collagen vascular disease (IPCVD) and alveolar type of HTLV-I associated bronchiolo-alveolar disorder (HABA-A) were evaluated. Soluble IL-2R (sIL-2R) in the lymphocyte culture supernatant increased in IIP, while the sIL-2R in sera and IL-2R positive lymphocyte showed no increase. In HABA-A, the serum sIL-2R level, the responsiveness to IL-2 and percentage of IL-2R positive lymphocytes and helper-T lymphocytes were increased. In IPCVD, the serum sIL-2R level increased but the percentage of IL-2R positive lymphocytes and helper-T lymphocytes decreased. The CD4/CD8 ratio was low in IIP and IPCVD, but high in HABA-A. In patients with IIP or IPCVD treated with steroids, the percentage of IL-2R positive lymphocytes and helper-T lymphocytes decreased more than in those not treated with steroids. Therefore, cellular immunity regulated by the IL-2 and IL-2R system might play an important role in the pathogenesis of IPCVD and HABA-A.
The serum eosinophil chemotactic factor (ECF) was evaluated by use of the modified Boyden chamber method. Flow cytometry (FCM) was applied to obtain highly purified eosinophils from peripheral blood of normal subjects. The maximal chemotaxis of these eosinophils was shown at 1×10-7 M of platelet activating factor and leukotriene B4. In bronchial asthmatics with immediate asthmatic response (IAR), serum ECF activity showed only an early rise, and that in asthmatics with late asthmatic response (LAR) showed a precedent rise before bronchial reaction. Asthmatic patients with both IAR and LAR showed biphasic elevation in ECF activity. These findings suggest that the function of ECF in the pathogenesis of IAR is different from that in LAR. The eosinophils purified by FCM were very useful for measurement of ECF by the Boyden chamber method.
The role of the eosinophil chemotactic factor (ECF) produced by peripheral blood mononuclear cells in the pathogenesis of bronchial asthma was studied. Bronchial asthma in childhood is characterized by its atopic history and such an asthmatic attack is induced by house dust or mite antigen through the type I allergic reaction. The lymphocytes found in bronchoalveolar lavage fluid have high responsiveness to Candida antigen. This suggests the participation of the type IV allergic reaction in the pathogenesis of asthmatic attack. Eosinophilic granulocyte infiltration in the lungs during the asthmatic attack was evaluated by measuring ECF activity produced by peripheral blood mononuclear cells incubated with Candida antigen. Chemotactic and chemokinetic activities to eosinophilic granulocytes were present in the supernatant, and basophilic granulocyte deletion did not change such chemotactic activity. High ECF production was observed in non-atopic asthmatics and in severe but non-steroid-dependent asthmatics. ECF may be produced at least in part from the lymphocyte blastogenic response to Candida antigen and may have some effect on the pathogenesis of non-atopic and severe asthmatics.
The DNA prepared from Vero cells infected with herpes simplex virus type 1 (HSV-1) was analyzed by pulsed field gradient gel electrophoresis (PFG). PFG was performed under three conditions using field inversion gel electrophoresis or orthogonal field alternation gel electrophoresis methods. Southern blot analysis with cloned HSV-1 DNA fragments was attempted for identification of HSV-DNA. The major part of HSV-DNA comigrated with HSV virion DNA, and was identified as HSV genomic DNA. Approximately 10% of HSV-DNA migrated a shorter distance than the HSV genomic DNA. The short migrating component consisted of one to three discrete bands depending on the condition of PFG. The molecular size of these bands were estimated to be more than 200 kilobase pairs. The short migrating component showed a tendency to comigrate with degradated cellular DNA, suggesting close association of the short migrating component with host cell DNA during viral replication. The mobility of the short migrating component was influenced by the treatment with several antiviral agents. Demonstration of the short migrating component of HSV-DNA by PFG may be direct evidence of the presence of high-molecular-weight replicative intermediates, and these observations are consistent with the “rolling circle model” in which the replicative intermediates are present as a concatemeric form.
The profiles of DNA fragments, generated by a restriction endonuclease BamHI, in the unique regions of herpes simplex virus type 1 (HSV-1) were compared among seven substrains originally derived from the parental Miyama strain. These substrains consist of five CPE variants (-GCr, -GCf, +GC (LPV), +GC (SPV) and +GC (81)) and two descendent strains of -GCr (αV and βV), both recovered from persistently infected L cell cultures originally established with -GCr. Some heterogeneous fragments were recognized among the seven substrains. The Southern blot hybridization analysis, using the subcloned DNA fragments of -GCr as probes, revealed the loss of the cleavage site between BamHI-T and BamHI-J' fragments in one of these substrains, αV. In vitro viral reproduction in an enormous number of cycles, which took place for a long period of 14 years in a persistently infected culture, might cause occasional spontaneous mutations of the virus carried in this culture and result in the loss of the cleavage site in the unique region of its genome.
The positively charged fluorescent dye Rhodamine 123 (Rho 123) accumulates in mitochondria. An adriamycin (ADR)-resistant cell line derived from Ehrlich ascites tumor cells (wild EATC) was established in our laboratory. Overall mitochondrial activity of wild EATC and ADR-resistant EATC was investigated by the staining method with Rho 123. The uptake of Rho 123 into living cells was analyzed by flow cytometry. The intracellular Rho 123 uptake of ADR-resistant EATC was lower than that of wild EATC. The intracellular Rho 123 uptake of wild EATC increased when cells were treated with cepharanthine (CP). The intracellular Rho 123 uptake of ADR-resistant EATC increased markedly by CP treatment. ADR-resistant EATC cosumed more endogenous oxygen than wild EATC. The energy level of ADR-resistant EATC was discussed.
To clarify the enzymatic properties of catalase from the acatalasemic mice, the change in the enzymatic activity during maturation of reticulocytes was observed both in normal and anemic conditions. Catalase activity in the blood of acatalasemic mice retained its maximum level 5 days after final injection of phenylhydrazine. An intimate lag correlation coefficient was obtained between reticulocyte count and catalase activity in blood by one day shift in acatalasemic mice and three days shift in normal mice, which corresponded to the interval between maximum peaks of reticulocyte count and catalase activity. The erythrocytes and reticulocytes were also counted, and the catalase activity per erythrocyte and reticulocyte in the blood of normal and acatalasemic mice was calculated. The catalase activity in blood due to reticulocyte retained its maximum level 5 days after injection of phenylhydrazine in normal and acatalasemic mice. The results revealed that catalase activity in acatalasemic mice was very low in reticulocyte and yet its degeneration by maturation was considerably great. Catalase activity per reticulocyte in the blood of acatalasemic mice was 55% of that of normal mice. Degeneration ratio expressed as (catalase activity in reticulocytes-that in erythrocytes)/catalase activity in reticulocytes was 88% in acatalasemic mice and 34% in normal mice. The difference in catalase activity between reticulocytes and erythrocytes in the blood of acatalasemic mice was also higher than that of normal mice. Thus the catalase activity in erythrocytes in the blood of acatalasemic mice was 10% of that of normal mice.