[Sensitivities of host cells for virus isolation from adenoviral conjunctivitis and DNA cleavage pattern analysis of adenoviruses by cell].
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Biomedical subjects
Publications and source records attributed to M Higuchi.
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Activities of angiotensin-converting enzyme (ACE), other proteinases, and eosinophil chemotactic factor (ECF-G) are known to be elevated in hepatic hypersensitivity granulomas of thymus intact (nu/+) mice after Schistosoma mansoni infection. The enzyme activities also increase, but to a lesser degree in hepatic granulomas of athymic nude (nu/nu) mice, and ECF-G is not detectable. In this study isolated hepatic granulomas from nu/+ mice were grafted into the skin of uninfected nu/nu mice, and changes in those cellular functions were determined to examine whether the newly formed granulomas by recipient nu/nu cells acquire the functional activities as well as the histological appearance of nu/+ granulomas. ACE and ECF-G rapidly disappeared from grafted sites during the first 5 days, corresponding to loss of nu/+ cells from the graft. Reduction in activities of arylsulfatases, lysozyme, and acid phosphatase also occurred, but to a lesser extent. Recovery of ACE and ECF-G activities to the levels seen in nu/+ hepatic granulomas was observed by 14 days after grafting when nu/nu cells had accumulated in the grafts and formed new granulomas. Other enzymes increased to approximately half the levels seen in grafted donor granulomas. Circulating eosinophilia also increased. The findings indicate that nu/nu cells that accumulated in the skin grafts not only morphologically mimicked nu/+ type granulomas but also demonstrated nu/+ levels of cellular function. Analysis of skin granulomas developing in nu/+ mice after grafting of nu/+ hepatic granulomas showed the similar histology and enzymatic changes, whereas the skin sites inoculated with purified schistosome eggs alone caused neither significant histological changes nor elevation of ACE activity.
Two hundred and thirty five subjects from 48 German cystic fibrosis (CF) families were typed for restriction fragment length polymorphisms (RFLPs) detected by the probes pmet H, pmet D, and pJ 3.11, known to be tightly linked to the CF gene. Gene and haplotype frequencies suggest a linkage disequilibrium with the CF locus. The analysis of the predictive value of this typing in individual CF families indicates that the combined use of these probes provides a powerful diagnostic system both for carrier detection and prenatal diagnosis. In 33 out of 48 families carriers and non-carriers could be identified, and in 26 of these 33 families prenatal diagnosis could discriminate between affected and unaffected offspring.
The influence of norepinephrine on lung fluid flow rate was studied in seven chronically catheterized fetal lambs. Lung fluid was collected through a catheter inserted into the fetal trachea. In six experiments, continuous saline solution infusion for 1 hour followed by equivolumnar norepinephrine (3.0 micrograms/min) for 1 hour via the fetal jugular vein was carried out. In three additional experiments (two fetuses), a 3.0 mg intravenous bolus of propranolol was administered before norepinephrine infusion. Saline solution infusion did not influence the lung fluid flow rate or fetal biophysical and biochemical parameters. Norepinephrine increased fetal blood pressure by 22% and decreased lung fluid flow by 45% from control values. Lung fluid flow rate recovered within 2 hours after the end of the norepinephrine infusion. Pretreatment with propranolol did not alter the results. In addition to previous reports that demonstrated beta-sympathomimetic agonist-enhanced reabsorption of lung fluid from fetal alveolar epithelial cells, our present results suggest that a reduction in fetal lung fluid secretion may be implemented as well through stimulation of alpha-sympathomimetic receptor sites in the pulmonary vasculature.
Human T cell hybridoma, H3-E9-6, that produces macrophage activating factors for cytotoxicity (MAF-C) was prepared by somatic fusion of phytohemagglutinin-activated peripheral blood lymphocytes with emetine/actinomycin D-treated cloned human acute lymphocytic leukemia cells (CEM 11). The activities of the following were assayed: (1) macrophage-activating factor for cytotoxicity of monocytes (MAF-C 1 day), (2) macrophage-activating factor for cytotoxicity of monocyte-derived macrophages (MAF-C 6 day), (3) macrophage-activating factor for cytotoxicity of murine macrophages (MAF-Cm), (4) macrophage-activating factor for glucose consumption (MAF-G), (5) macrophage-activating factor for O2- formation (MAF-O). The culture supernatant of H3-E9-6 showed MAF-C 1 day-MAF-C 6 day, MAF-Cm, and MAF-G activities. The MAF-Cm activity was considerably enhanced by the addition of murine recombinant interferon gamma (rIFN-gamma). The MAF-C 1 day activity in the H3-E9-6 sup was not decreased by heat treatment (56 C, 30 min), by pH 2 treatment or by the addition of monoclonal anti-human IFN-gamma antibody or polymyxin B. These data suggest that MAF-C in H3-E9-6 sup is distinct from human IFN-gamma or lipopolysaccharide (LPS).
Two different factors (MAF-C I and MAF-C II) were obtained by anion exchange chromatography of the culture supernatant of a human T-cell hybridoma, H3-E9-6, which produces macrophage-activating factors for cytotoxicity (MAF-C). These 2 factors induced the cytotoxicity of monocytes synergistically as a priming signal (MAF-C I) and a triggering signal (MAF-C II), respectively. On gel filtration on a column of Superose 12, MAF-C II was eluted mainly at the void volume, whereas MAF-C I was eluted in the fractions corresponding to approximate molecular weights of 30-300 K. On the other hand, gel filtration in the presence of sodium deoxycholate revealed that MAF-C II has an approximate molecular weight of 40,000, but MAF-C I was unstable under these conditions. When the activity for mouse macrophages (MAF-Cm activity) was tested, the MAF-C II fraction showed high MAF-Cm activity in the presence of murine recombinant interferon gamma (rIFN-gamma), but the MAF-C I fraction did not show MAF-Cm activity even in the presence of lipopolysaccharide (LPS). These results suggest that MAF-C I (priming lymphokine) has species specificity but MAF-C II (triggering lymphokine) does not.
Effects of nipradilol on hemodynamics and transmural energy metabolism of underperfused (ischemic) canine hearts were investigated. The ischemic heart was prepared by constricting a tube connecting the circumflex coronary artery with the carotid artery for 10 min so that the perfusion pressure (CPP) was reduced to 30 mmHg. The reduction in CPP resulted in decreases in coronary blood flow (CBF) by 70%, regional myocardial contractile force (MCF) by 30%, myocardial ATP contents by 32% (inner layer)-22% (outer) and creatine phosphate by 75% (inner)-60% (outer). Increases in the left ventricular end diastolic pressure (LVEDP) by 4.8 mmHg, myocardial inorganic phosphate contents by 1.9 times (inner)-1.3 (outer) and lactate by 4.3 times (inner)-2.4 (outer) were also observed. In dogs with normal hearts, an infusion of nipradilol (10 micrograms/kg/min, i.v., for 15 min) decreased CPP by 25%, CBF by 40%, cardiac effort index by 45% and MCF by 30 to 40%, and it slightly increased LVEDP without affecting myocardial high-energy phosphate and lactate levels. In ischemic hearts, nipradilol infusion starting 5 min before ischemia attenuated the ischemia-induced elevation of LVEDP to 1.8 mmHg, and the ischemia-induced changes in high-energy phosphate contents to 1/2 (inner)-1/3 (outer) and changes in lactate to 1/6 (inner)-1/10 (outer). These results indicate that nipradilol improves the ischemic derangement of both transmural energy metabolism and hemodynamics.
DNA restriction endonuclease cleavage analysis and serum neutralization test were performed in strains of adenovirus types 19 (Ad-19) and 37 (Ad-37) isolated from patients with acute conjunctivitis in Sapporo (Japan), Kaohsiung (Taiwan) and Pusan (South Korea) during the period of 1977 to 1984. Ad-19 and Ad-37 could be roughly differentiated, but a fair degree of cross reactivity was noted by serological test. By DNA restriction endonuclease analysis, Ad-19 isolates, Ad-19 prototype strain and Ad-37 strains were clearly differentiated. It was suggested by DNA cleavage patterns that both Ad-19 and Ad-37 isolates in East Asia are the same as those currently found in Europe and the USA. A new subtype of Ad-37 was found by DNA cleavage analysis among the isolates obtained in Sapporo and Kaohsiung in 1980.
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We prepared large unilamellar vesicles containing the cell-free culture supernatant of a human T cell hybridoma rich in macrophage activating factor (MAF) and bearing monoclonal antibodies against human melanoma A375 tumor cells; their antitumor activity against A375 cells was examined in vitro and in vivo. Both MAF-immunoliposomes (bearing antibodies) and MAF liposomes (not bearing antibodies) showed macrophage-mediated cytotoxicity in vitro at a high E/T ratio (about 40). But at a low E/T ratio (about 15), only MAF-immunoliposomes showed tumoricidal activity, their activity being more than ten thousand-fold stronger compared with a soluble MAF preparation (MAF solution). MAF-immunoliposomes not only showed tumor neutralization mediated by macrophages in vivo when a mixture of tumor cells, macrophages, and MAF-immunoliposomes was locally injected, but also showed significant inhibition of tumor growth on repeated i.v. systemic administration of them. On the other hand, other samples (MAF-liposomes without the antibody, a MAF solution, and immunoliposomes without MAF) were not significantly effective against tumor growth. These results may constitute evidence that the delivery of lymphokines to the tumor sites is important or even critical when an attempt is made to treat cancer with lymphokines with the expectation of the potentiation of the host's immune system.
The concentrations of disaturated phosphatidylcholine (DSPC), the major component of pulmonary surfactant, were quantified on 528 amniotic fluid samples, which did not show the contamination of blood or meconium and were uncomplicated by polyhydramnios or severe congenital anomalies incompatible with life. 396 samples of amniotic fluid (29 RDS cases) obtained 72 hours before delivery were used to evaluate the reliability of DSPC as a specific indicator in pedicting fetal lung maturation. DSPCs were determined according to the method described by Mason et al. and lecithin/sphingomyelin (L/S) ratios were measured by using two dimensional thin-layer chromatography. The critical points of DSPC and L/S ratio were defined as 1.0mg/dl, 2.0 respectively. The results are as follows. There was a gradual increase in DSPC levels of amniotic fluid with increasing maturity until 35 weeks of gestational age and a rapid rise from 36 weeks to term. About 70% of the amniotic fluids having a significantly higher DSPC level greater than or equal to 5.0mg/dl within 35 weeks of gestation were found to be obtained from pregnant women whose pregnancy was complicated by PROM, threatened premature labor or cervical incompetency and who eventually underwent preterm labor. Out of 58 samples, in which DSPC and the L/S ratio were measured simultaneously, two markers agreed in 47 cases in an immature or a mature value for predicting fetal lung maturation. RDS was correctly predicted in 27 of 60 cases (false negative, 55.0%) with DSPC levels less than 1.0mg/dl. When DSPC levels were 1.0mg/dl or more, there were only two of 336 (0.6%), false positives.(ABSTRACT TRUNCATED AT 250 WORDS)
We have studied the effects of reoxygenation on hypoxia-induced changes in contents of high energy phosphate compounds in pacemaker cells of nodal tissues excised from the rabbit heart, and effects of coenzyme Q10 on the electrical activity and metabolite contents in the tissue exposed to hypoxia and then reoxygenated. The contents of ATP and creatine phosphate (CP) in the sinoatrial node tissue were markedly reduced within 15-30 min after exposure to hypoxic Tyrode's solution. Reoxygenation produced almost complete recovery of the tissue ATP, but not of CP. After 60-120 min of hypoxia, the tissue ATP and CP decreased to about 60-30% of the initial value, but were not recovered by reoxygenation. Coenzyme Q10 (CoQ) at concentrations of 10(-6)-10(-5) g/ml did not produce changes in action potential parameters in the normal Tyrode's solution. CoQ (10(-5) g/ml) did not prevent the decreases in tissular ATP and CP in the initial period of hypoxia (30-60 min), but the ATP content at 120 min of hypoxia in the presence of CoQ was higher than the control. CoQ promoted recovery of tissular ATP after reoxygenation. Our results provide direct evidence that generation of action potentials in the sinoatrial nodal cells can be maintained by a small amount of ATP produced by the anaerobic glycolytic pathway. The present results suggest that exogenous CoQ would facilitate resynthesis of ATP in the functionally impaired mitochondria.
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