Interaction between dyes and polyelectrolytes, 8. Effect of polyanions on the mixed dimer formation of cationic dyes

The effect of polyanions on the formation of mixed dimers of methylene blue ( 1 ) and trypaflavine ( 2 ), methylene blue ( 1 ) and phenosafranine ( 3 ), and methylene blue ( 1 ) and pyronine G ( 4 ) was investigated spectrophotometrically. The following polyanions were used: poly(potassium styrenesulfonate) (PSS), poly(potassium vinyl sulfate) (PVS), and poly(sodium acrylate) (PAA). On addition of polyanions, the formation of mixed dimers was enhanced largely. Thermodynamic parameters inferred that the enhancement of the formation of mixed dimers in the presence of polyanions resulted from an entropic factor.     

Interaction between dyes and polyelectrolytes, 2. Structural effect of polyanions on the methylene blue binding

The structural effect of polyanions on the binding type of methylene blue ( 1 ) was investigated spectrophotometrically. 1 was bound to poly(potassium styrenesulfonate) (PSS) and poly(sodium 4-vinylphenylsulfate) (SVS) in the dimeric or slightly aggregated form and to poly(sodium vinylsulfonate) (SVF) and poly(potassium vinyl sulfate) (PVS) in the highly aggregated (polymeric) form. It was found that the flexibility of polyanions plays an important rǒle in the aggregation of bound 1 and that the difference between ? SO and ? OSO as binding site is not a significant factor.     

IMMUNOHISTOCHEMICAL STUDY OF PANCREATOBLASTOMA

Three cases of pancreatoblastoma in children were examined immunohis-tochemically and the results were compared with those of pancreatic duct carcinoma in adults. The pancreatoblastoma demonstrated positive reactions to α-fetoprotein (AFP) (67%: 2/3), α-1-antitrypsin (AAT) (100%: 3/3), carcinoembryonic antigen (CEA) (67% : 2/3) and keratin (33% : 1/3), although CEA was only weakly positive in both cases. On the other hand, adult pancreatic duct carcinoma showed positive reactions as follows; AFP: 3% (1/29), AAT: 21% (6/29), CEA: 97% (28/29) and keratin: 93% (27/29). Also, endocrine substances including insulin, glucagon and somatostatin were all negative in the pancreatoblastomas. Two cases of pancreatoblastoma which were immunohistochemically positive for AFP also showed elevation of the serum AFP level clinically. The different expressive pattern of oncofetal antigens in pancreatoblastoma as compared with pancreatic duct carcinoma in adults may provide further supporting evidence for the embryonic nature of pancreatoblastoma, and suggests that such a pattern might be used as a tumor marker for pancreatoblastoma. ACTA PATHOL. JPN. 37 : 1581-1590, 1987.     

Expression of heme oxygenase-1 in rat ontogeny

Heme oxygenase (HO), the heme-degrading enzyme, plays an important role in heme catabolism. Among three isozymes, HO-1 is an inducible form expressed mainly in macrophages. In rat ontogeny, HO-1 immunoreactivity was detected in mononuclear cells in the yolk sac at 10 days of gestation. HO-1-expressing cells were then detected in the fetal liver and their numbers increased during the gestational period. The numbers of HO-1-positive cells and HO-1 mRNA levels in the liver peaked at 18 days of gestation. Most of the macrophages expressed both HO-1 and a macrophage scavenger receptor. Macrophages in the fetal liver showed marked hemophagocytosis. Macrophages in the lung, spleen, bone marrow, and other tissues also expressed HO-1. HO-1 immunoreactivity was also observed in syncytial cells of the chorionic villi, the endodermal layer of the yolk sac, and renal tubules of the fetus. Intestinal mucosal epithelial cells expressed HO-1 after birth. These findings imply that HO-1 is crucial for macrophages in heme catabolism from an early stage of ontogeny. HO-1 expression in non-macrophagic cells may be required for other purposes such as protection from oxidative stress and various stimuli.     

Three-dimensional two-layer collagen matrix gel culture model for evaluating complex biological functions of monocyte-derived dendritic cells

Dendritic cell-like cells (Mo-DCs) generated from peripheral blood monocytes with interleukin-4 (IL-4) and granulocyte-macrophage colony-stimulating factor (GM-CSF) have been used as tools to treat cancer patients (DC-vaccines). Because Mo-DCs have multiple antigen presentation-related functions, including phagocytosis, migration, cytokine production, and T cell stimulation, establishment of a method for simultaneously evaluating the various functions of Mo-DCs is important. We developed a new in vitro three-dimensional two-layer collagen matrix culture model that consists of a collagen gel containing Mo-DCs as the lower layer and a collagen gel containing necrotic GCTM-1 tumor cells and/or T cells as the upper layer. We used this system to observe simultaneously multiple functions of Mo-DCs by phase-contrast or fluorescence microscopy and to assess IL-12 secretion during more than 2 weeks of culture. We also observed interactions between Mo-DCs and necrotic GCTM-1 or T cells on an individual cell basis by time-lapse videomicroscopy. In addition, we collected Mo-DCs from the collagen gels by collagenase treatment and analyzed the expression of antigen presentation-related molecules such as HLA-DR, CD80, CD83, and CD86 on Mo-DCs. This model may be a useful tool for evaluation of the various functions of Mo-DCs used as DC vaccines and for studies of the complex behaviors of Mo-DCs in vivo.     

Role of KCNQ1 in the cell swelling-induced enhancement of the slowly activating delayed rectifier K(+) current

Cell swelling enhances a slowly activating delayed rectifier K(+) current (I(Ks)) in cardiac cells. This investigation was undertaken to determine which of the two structural units reconstituting the I(Ks) channel, KCNQ1 (KvLQT1) and KCNE1 (minK/IsK), plays a key role in the cell swelling-induced I(Ks) enhancement and to dissect a possible involvement of tyrosine phosphorylation therein. KCNQ1 was transiently expressed alone or together with KCNE1 in a heterologous mammalian cell line. Two distinct whole-cell membrane currents were separately observed during the exposure of transfected cells to various degrees of hyposmotic solutions. A hyposmotic challenge (0.7 times control osmolarity) resulted in about a twofold increase not only in the heteromeric KCNQ1/KCNE1, but also in the homomeric KCNQ1 channel currents. There was no significant difference in the incremental ratio of current amplitude in response to hyposmotic stress between the two KCNQ1-related currents, and the cells expressing the heteromeric channels swelled less than those with the homomeric channels or without the exogenous ones. The cell swelling-induced I(Ks) enhancement was not affected by a protein tyrosine kinase (PTK) inhibitor, by genistein (50 microM), or by an inhibitor of phosphotyrosine phosphatase (PTP), orthovanadate (500 microM), or a nonhydrolyzable ATP analogue, AMP-PNP (5 mM). Taken together, it is very likely that KCNQ1 might primarily participate in the I(Ks) enhancement by osmotic cell swelling. The obligatory dependence of the I(Ks) augmentation on PTK activity remained to be demonstrated, at least, in this expression system.     

Organ design for generation and reception of CO: lessons from the liver

Carbon monoxide (CO) is synthesized in vivo by heme oxygenase. Although for many years CO had been regarded as potentially toxic waste, recent studies have indicated that it is a signaling molecule with important physiological functions. Nitric oxide (NO), another diatomic diffusible gas, is regarded as an established signaling molecule. Structural similarities between CO and NO have led many investigators to draw analogies between the two gaseous mediators. Whereas the NO signaling system has been well defined as to its receptor molecule, soluble guanylate cyclase, the CO system has been conceived to require further tuning with respect to identifying its receptor molecules and its downstream effectors. Furthermore, there has been little quantitative information to argue for a physiological role of CO in vivo. This review, therefore, focuses on recent developments on both physiologic and pathophysiologic roles of CO in the model of isolated perfused liver of rats where endogenous production of CO is actually estimated. This model has revealed that CO acts as an endogenous vasorelaxant in the liver and that effects of CO are at least in part cyclic GMP-dependent. It has also provided answers to many questions of hepatobiliary functions that had not been resolved because of the complexity introduced by the interplay between NO and CO.     

Effects of fluid flow on elution of hydrophilic modifier from dialysis membrane surfaces

When uremic blood flows through dialyzers during hemodialysis, dialysis membrane surfaces are exposed to shear stress and internal filtration, which may affect the surface characteristics of the dialysis membranes. In the present study, we evaluated changes in the characteristics of membrane surfaces caused by shear stress and internal filtration using blood substitutes: water purified by reverse osmosis and 6.7 wt% dextran70 solution. We focused on the levels of a hydrophilic modifier, polyvinylpyrrolidone (PVP), on the membrane surface measured by attenuated total reflectance Fourier transform infrared spectroscopy. Experiments involving 4 h dialysis, 0-144 h shear-stress loading, and 4 h dead-end filtration were performed using polyester-polymer alloy (PEPA) and polysulfone (PS) membranes. After the dialysis experiments with accompanying internal filtration, average PVP retention on the PEPA membrane surface was 93.7% in all areas, whereas that on the PS membrane surface was 98.9% in all areas. After the shear-stress loading experiments, PVP retention on the PEPA membrane surface decreased as shear-stress loading time and the magnitude of shear stress increased. However, with the PS membrane, PVP retention scarcely changed. After the dead-end filtration experiments, PVP retention decreased in all areas for both PEPA and PS membranes, but PVP retention on the PEPA membrane surface was lower than that on the PS membrane surface. PVP on the PEPA membrane surface was eluted by both shear stress and internal filtration, while that on the PS membrane surface was eluted only by internal filtration.