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We have already developed the liposome immune lysis assay (LILA) for the determination of C-reactive protein (CRP) by employing an inhibition method and a sandwich method. We herein report a new LILA system involving the use of monoclonal antibodies-bearing liposomes. We established five monoclonal antibodies to CRP antigen, AC-1, -2, -3, -4, -5 which had the capacity to activate complement and form antigen-antibody complex. Each of these antibodies was covalently coupled to carboxyfluorescein-entrapped multilamellar liposomes. When the liposomes were incubated with CRP antigen in the presence of guinea pig complement, CRP antigen-dependent liposome lysis was observed but the sensitivity was not great enough for practical use. On the other hand, when liposomes coupling two monoclonal antibodies (AC-1, AC-2) which recognized distinct CRP antigenic determinants were employed in the assay, the sensitivity increased compared with that using only one monoclonal antibody, and the detectable concentration range was 5-300 ng/ml. These results indicated that the combination of two or more monoclonal antibodies which recognize distinct CRP antigenic determinants is effective for increasing the sensitivity of the assay.

Hepatitis C virus (HCV)-RNA in the blood was measured by polymerase chain reaction (PCR) in 37 subjects from eight families in which 2 or more persons tested seropositive for antibodies against C100-3 or CP9. HCV-RNA was positive in 17 of 37 subjects. Two or more HCV-RNA-positive subjects were observed in six of the families. Intrafamilial HCV infection was studied by determining the HCV-RNA type (I, II, III or IV) by PCR using type-specific primers. In two families, all of the subjects showed type III infection, and in three other families, all of the subjects showed type II infection, with different types of HCV infections being observed in only one family. The HCV type was uniform in all but one. These findings suggest a possibility of intrafamilial infection between husbands and wives and between members of the same household.

A unilateral 30-min application of anodal direct current to the promotor cortex of rabbits was repeated 10 times, and cyclic AMP accumulation in response to histamine was investigated in slices of different cortical areas. Polarization with 1.0 microA decreased the cyclic AMP accumulation in the cortical area contralateral to the polarization, by which regional dominance in cyclic AMP accumulation was produced in the polarized cortex. In contrast, the regional difference in cyclic AMP accumulation was reversed when 10.0 or 30.0 microA was applied. The histamine-elicited accumulation of cyclic AMP was almost completely inhibited by the selective H2-receptor antagonist cimetidine. These results suggest that repeated anodal polarization regionally alters H2-receptor-mediated cyclic AMP generation in the cortex depending on the intensity of the polarizing currents and this pattern of cyclic AMP accumulation is responsible for the characteristic motor behavior induced by anodal polarization.

We studied the distribution of class 1 and class 2 major histocompatibility complex (MHC) antigens on bile duct epithelial cells in liver from patients with primary biliary cirrhosis (PBC) by an immunohistochemical method using monoclonal antibodies to HLA-ABC products and HLA-D subregion products (HLA-DR, -DP, -DQ). By light microscopy, the expression of MHC class 1 antigens (HLA-ABC antigens) was enhanced in PBC compared with controls. While negligible staining of MHC class 2 antigens was detected on the bile duct in controls, de novo expression of MHC class 2 antigens, as well as the coexpression of HLA-DR, HLA-DQ, and HLA-DP antigens on the bile duct epithelial cells, was observed in PBC. By electron microscopy, HLA-ABC and HLA-DR antigens were present preferentially along the basolateral domain of the cell surface of the bile duct epithelial cells and on the membrane of the endoplasmic reticulum in the cytoplasm, suggesting that these MHC antigens are synthesized by the bile duct epithelial cells in PBC. The distribution of these MHC antigens on the basolateral surface of the bile duct epithelial cells, where they are easily accessible to immunocytes, supports the idea that MHC-restricted cytotoxic T lymphocytes are involved in the bile duct injury in PBC.

The aim of this study was to investigate the usefulness of magnetic resonance (MR) imaging in the evaluation of renal function, with particular attention to the effects of water load. Ten healthy volunteers underwent dynamic MR imaging after an injection of gadolinium diethylene-triaminepenta-acetic acid (Gd-DTPA) as a contrast agent to evaluate renal function by the following four methods: the positive method [longitudinal relaxation time (T1) shortening is the dominant effect], the negative method [transverse relaxation time (T2) shortening is the dominant effect] and two intermediate methods by switching the Gd-DTPA concentrations used in the positive and negative methods. A prolonged cortical peak time and a reduced medullary peak level were observed by the positive method under a dehydrated condition, suggesting that these variables were slightly influenced by Gd-DTPA concentrated in the medulla. By the negative method, low signals due to T2* (T2* is the effective transverse relaxation time, typically shorter than T2) shortening appeared in the medulla under normal conditions, but these signals were unclear when the subject was under an overhydrated condition. These results indicate that water metabolism, in addition to imaging parameters and Gd-DTPA dose levels, should be considered when renal function is evaluated by dynamic MR imaging. Analysis of both the pattern of MR images and the time-signal intensity curves may be useful in the evaluation of renal function. The results also indicate that the positive method is preferred when the patient is overhydrated as it allows the evaluation of the local renal kinetic function by recording changes in the regional contrast agent levels.

In this study, we investigated serum-soluble interleukin-2 receptor (sIL-2r) and neopterin (NPT) levels in five patients with severe postoperative infections. A total of 25 synchronous determinations of sIL-2r and NPT were performed. A marked increase in sIL-2r and NPT levels was observed, and the increase in sIL-2r was significantly correlated to that of NPT which is a marker of macrophage activity. These results suggest that macrophages are involved in the stimulation of sIL-2r release, representing a potentially negative biological effect. The results indicate that sIL-2r may be a useful indicator of the efficacy of antibiotics and of prognosis.

Both prolidase and prolinase from the human prostate were separated into two peaks by TSK DEAE-5PW chromatography. These peaks of prolidase isozymes I and II differed from each other in their responses to preincubation with Mn2+, their substrate specificity, optimal pH, and heat stability. The molecular weights of prolidases I and II were estimated to be 110,000 and 165,000, respectively, by gel filtration. Substrate specificity of prolinase peaks I and II was almost the same, but they differed in optimal pH and heat stability. The molecular weights of prolinases I and II were about 85,000 and 63,000, respectively. These results indicate that two isozymes of prolidase and of prolinase, which differ in various characteristics, are present in the human prostate.

Neurons with strongly negatively charged surface coats were recognized in mammalian, avian, reptilian, amphibian and piscine brains. Many large-sized neurons had strongly negatively charged surface coats in the visual cortex and brain stem of the cow, cat, guinea pig, mouse, quail and parakeet. Such neurons were also seen in the brain stem of the lower vertebrates such as the house lizard, Japanese terrapin, bullfrog, newt, carp and sweetfish.

The sensitivity and specificity of single cell polymerase chain reaction (PCR) were studied. Its high sensitivity enabled detection of a single-copy gene, such as human T-lymphotropic virus type I genome in paraffin sections. The rate of obtaining positive signals with this method was affected by the number of copies of the gene in the target cell. Specificity was satisfactory if the procedure was properly and carefully followed. Since the single cell PCR is a time-consuming method which requires skill and experience to pick up the target cells accurately, the applicability of this method is limited. It works best when it is used to analyze a single or a few copy genes in histologically identified cells.

Familial adenomatous polyposis (FAP) is a well-known autosomal dominant disorder characterized by the formation of multiple adenomatous polyps of the colon. Gardner's syndrome is a variant of familial polyposis coli, and both can be associated with colonic or extracolonic benign and/or malignant tumors. It has been widely recognized that an adenocarcinoma of the colon develops in virtually all cases, usually at an earlier age, if polyps are left untreated. Families of four individuals diagnosed of FAP were surveyed and 56 relatives of the families were examined. Of these 56, 21 had multiple colon polyps, 3 of whom had early-stage adenocarcinomas. We consider that familial survey of FAP individuals can be of considerable benefit for this high-risk population due to the autosomal nature of the disease, allowing diagnosis of an associated cancer at an earlier stage.

The accumulation of polymorphonuclear leukocytes (PMN) in synovial fluid is a common feature of rheumatoid arthritis (RA). We studied the chemotactic response of PMN obtained from the synovial fluid and from the peripheral blood of patients with RA using a modified Boyden's method, in which interleukin-8 (IL-8) or N-formyl-methionyl-leucyl-phenylalanine (FMLP) was used as a chemotactic agent. The IL-8-induced response of peripheral blood PMN from 15 patients with RA did not differ from that of 15 healthy controls. A decreased chemotactic response to IL-8 was, however, observed in PMN from the synovial fluid of 12 patients with RA compared with peripheral blood cells of the same individual. This defective chemotactic ability of PMN was inversely correlated with the number of infiltrating cells in the synovial fluid. We also obtained similar results with FMLP. These results indicate that the chemotactic ability of PMN may be reduced after migrating to the synovial fluid.

To clarify the influence of the introduction of metal bats on the physique and batting records of the players in the National Summer High School Baseball Tournaments, a comparative analysis was conducted between height, body weight, body-weight ratio (body weight/height), batting average and home run average of the best four teams' players (n = 493) and those of the other teams' players (n = 4,590) in three periods: the period of the use of wooden bats, that of the use of both wooden and metal bats and that of the use of metal bats. In the period of metal bat use, the mean values of physique of the best four teams' players were significantly larger (P < 0.05) and their average number of home runs was significantly higher than those of the other teams' players (P < 0.01). The only significant differences between the two groups in those indices for each time period were for height in the wood and metal/wood periods. This study demonstrated that the differences between the home run average and physique, including height, body weight and body-weight ratio of the best four teams and the rest of the teams were the greatest after the use of metal bats. These findings suggest that the importance of the home run average increased and was associated with large-size of physique after the use of metal bats in the National Summer High School Baseball Tournaments.

External capacitive heating is the usual method of electromagnetic wave heating, in which the tumor is caught and heated between two opposite applicators. Using a phantom, the authors developed and evaluated the performance of a new capacitive heating applicator designed for simultaneous radiohyperthermotherapy (SRH) in which the electron beam irradiation is provided from above an external capacitive heating applicator for the treatment of superficial and shallow-seated tumors. The trial applicator was constructed to fulfill the following conditions: 1. use of an electrode plate which does not affect the electron beam depth dose, 2. a uniform thickness to maintain flatness of the electron beam, and 3. a cooling function to prevent damage to normal skin tissue and enhance the therapeutic gain factor. This applicator was comprised of a 0.1-mm-thick copper electrode and a 5-mm-thick cooling chamber. The depth of the 80% dose of the new applicator was 21 mm with a 9-MeV electron beam and 36mm with a 15-MeV electron beam, which was comparable to the effect of a conventional irradiation bolus. The temperature distribution produced by the trial applicator was symmetrical on both sides from the center of the applicator. The 50% specific absorption rate region was 6.4 cm wide at a depth of 1 cm from the phantom surface and 2.8 cm wide at a depth of 3 cm. There have been no previous reports on the development of an external capacitive heating applicator designed for the SRH of superficial and shallow-seated tumors; this is the first such report.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of ischemia and nitroglycerin on systolic time intervals in the segmental myocardial length were studied in anesthetized open-chest dogs. Two strain-gauges were sutured on the surface of the left ventricular wall; one was in the central area perfused by the left circumflex coronary artery (LCX) and the other was in the area perfused by the left anterior descending coronary artery. LCX was partially occluded with a screw type constrictor to the degree at which reactive hyperemia after the transient total coronary occlusion almost disappeared. After the hemodynamics stabilized nitroglycerin (20 microgram/kg) was injected into the femoral vein. In the ischemic area, contraction time was shortened and precontraction time was prolonged in association with an elongation of end-systolic and early systolic segment-length, respectively. The systolic time intervals in the ischemic segment were improved as a result of the recovery in the segment-length toward the control. The results suggest the usefulness of analyzing the segmental myocardial systolic time intervals for verifying the asynchronous contraction of the ventricle and the favourable effects of nitroglycerin on segmental myocardial function in the ischemic area.

A blood coagulation deficiency was found at the contact phase in identical Japanese female twins. Of the four possible factors involved, Factor XI or XII can be ruled out according to cross-correction studies. The problem factor was probably not Fletcher factor, because the abnormal partial thromboplastin time was not significantly shortened by increasing the incubation period of plasma with kaolin. The deficiency is most likely due to the lack of Fitzgerald factor.

Previous investigation showed that mouse ascites sarcoma cells permeabilized with appropriate concentrations of detergents (Triton X-100, Nonidet P-40 and Brij 58) had high replicative DNA synthesis in the presence of the four deoxyribonucleoside triphosphates, ATP, Mg2+ and proper ionic environment. The present study showed the optimum detergent concentration for DNA synthesis coincided closely with the minimum detergent concentration for inducing cell swelling. Phase contrast microscopy and electron microscopy of Triton-permeabilized cells showed the characteristic swollen cytoplasms and nucleus. Autoradiographic study showed that the DNA synthesis in permeable cells was confined to the nucleus. Cell viability and [3H] deoxythymidine uptake were impaired at much lower concentrations of Triton X-100 than the optimum concentration for in vitro DNA synthesis. In Triton-permeabilized cells, the minimum Triton concentration that produced cell swelling also seemed to produce high repliative DNA synthesis, which reflects the in vivo state of DNA synthesis.

Delayed cerebral vasospams is caused by excessive accumulation of dopamine-beta-hydroxylase (DBH) and noradrenaline in cerebral vessel walls. This study demonstrates the mechanisms of delayed spasm, particularly the role of red blood cell components, and the successful relief of delayed cerebral vasospasm. Spasmogenic substances which contained a heme component, such as methemoglobin, methemalbumin and catalase enhanced DBH activity in human serum as measured by a one step chemical spectrophotometric assay. The concentration which gave the highest DBH activity caused the maximum constriction of the basilar artery, when the substances were applied topically. Among components of red cells, methemoglobin, methemalbumin, catalase and nicotinamid adenin dinucleotide (NADH) caused constriction of basilar artery in cats, when applied topically, whereas hematin, hemin and bilirubin caused no significant spasm. An oxyhemoglobin solution obtained by mixture with methemoglobin and ascorbic acid produced no significant vascular spasm either. Relief of delayed cerebral vasospasm was obtained with topical application of specific alpha adrenergic blocking drug such as phenoxybenzamine, specific inhibitors of DBH such as fusaric acid, o-phenanthroline and alphaalpha' dipyridyl beta2 adrenergic stimulants such as salbutamol, and a phosphodiesterase inhibitor, ascorbic acid.

This study demonstrates that an adrenergic mechanism plays an important role in producing the delayed cerebral vasospasm which follows subarachnoid hemorrhage. Results were as follows: 1. Experimental subarachnoid hemorrhage (SAH) was produced by injection of fresh arterial blood into the cisterna magna in cats. The cerebral vasospasm was shown angiographically to be biphasic in nature: immediate constriction lasting 1 h and marked prolonged spasm occurring between the 3rd and 5th day after SAH. The amount of noradrenaline (NA) and dopamine-beta-hydroxylase (DBH) activity decreased over a period of 24 h both within the wall of the basilar artery and in the locus ceruleus and then gradually increased, reaching a maximum on the 3rd day after SAH. 2. Topical application of spasmogenic substances (NA and blood) produced a marked constriction of the hypersensitive basilar artery on the 3rd day after SAH. 3. 6-Hydroxydopamine (6-OHDA) injection into the cisterna magna produced prolonged vasocilatation. The dilated vessel responded with mild transient constriction after the topical application of NA or fresh blood. DBH activity and NA concentration in the vessels, locus ceruleus and medial hypothalamus decreased markedly on the 3rd day after the cisternal injection of 6-OHDA. 4. Various spasmogenic substances (i.e. serotonin, NA, prostaglandins and methemoglobin) were measured in a mixture of equal volume of CSF and blood in cats. ONly the serotonin in the mixed fluid produced vasoconstriction. Spasmogenic substances decreased markedly in the mixed fluid incubated for 3 days at 37 degrees C, and none of these substances apart from methemoglobin was present in a concentration sufficient to produce constriction of vessels. 5. These results suggest that early spasm is induced by serotonin around the arteries of the cranial base, and delayed spasm might be caused by hyperreaction of cerebral vessels to spasmogenic substances such as methemoglobin, during the accumulation of excess NA in the cerebral vessel wall.

Primary mass culture of isolated cells from adult rat livers by the trypsin-perfusion method uas carried out to investigate cytological and biochemical properties of primary cultured cells. Two main types of cells were found in the course of primary culture of isolated hepatic cells. One was a group of polygonal cells with granular cytoplasm, which appeared to arrange themselves in cords and clusters 3 to 4 days postinoculation and then gradually decreased in number. These cells seemed to be associated with expression of liver specific functions such as higher tyrosine aminotransferase levels and transient increases in albumin production. Another type of cells was relatively small in size and had clear cytoplasm, which grew out rapidly in place of the polygonal cells and proliferated also in subculture. alpha-Fetoprotein was detected in the cultured media of the proliferating clear cells. These results suggest that the polygonal cells and the clear cells may correspond to the mature hepatocytes and the hepatocytic stem cells in vivo, respectively.

We report the first case in Japan, i.e., the first case among oriental subject of Farber's disease. This is a rare disorder of lipid metabolism in infancy subsequent to a genetically-determined defect in ceramide degradation. Main features are characterized clinically by hoarseness, joint swelling, subcutaneous nodules and retarded psychomotor development. Lipid analysis and pathological investigation on the material obtained from a subcutaneous nodule confirmed clearly the presence of ceramide and intracytoplasmic inclusion bodies characteristic for Farber's disease. In this case, we experienced also corneal opacity and striking abnormalities in electroencephalogram, which have apparently not been noticed in the 17 cases hitherto reported.