Systemic adjuvant therapy for node-negative breast cancer.

In response to recent advice from the US National Cancer Institute concerning the use of systemic adjuvant therapy for node-negative breast cancer we reviewed the literature and found that several studies have shown evidence of a disease-free, but not an overall, survival advantage for treated patients. The benefits have been modest and may not outweight the cost and toxic effects of such therapy. Routine use does not seem to be justified. Factors must be identified to differentiate between patients at low risk and those at high risk. It should then be determined if adjuvant therapy is truly beneficial in those who are at high risk.     

Vascular endothelial cell killing by combinations of membrane-active agents and hydrogen peroxide

Previous studies have demonstrated that a number of membrane-active agents are capable of binding to the surface of polymorphonuclear leukocytes (PMN) resulting in an augmentation of superoxide anion and hydrogen peroxide (H2O2) production in response to soluble stimuli. It is now demonstrated that these same membrane-active agents can bind to the surface of endothelial cells and enhance their susceptibility to killing by H2O2. Membrane-active agents which are capable of synergizing with H2O2 include cationic proteins, cationic poly-amino acids, lysophosphatides and enzymes which are capable of degrading membrane phospholipids (e.g., phospholipase C, phospholipase A2 and streptolysin S). In each case, treatment of the target cells with the membrane-active agent and H2O2 produces greater damage than the sum of the damage produced by either agent separately. Since inflammatory lesions, particularly sites of bacterial infection, may contain a rich mixture of cationic substances, phospholipases and phospholipid breakdown products, these substances may contribute to the tissue damage observed at sites of inflammation by enhancing endothelial cell sensitivity to PMN-generated H2O2 as well as by augmenting the generation of H2O2 by PMNs.     

Ultrastructural aspects of the adherence to target cells of in vitro differentiated lymphocytes.

The specific adherence to target fibroblasts of lymphocytes differentiated in vitro from blast cells after stimulation with pokeweed mitogen was studied under the electron microscope. Lymphocyte pseudopods were seen to penetrate into the target cells forming a close contact between the membranes of the two interacting cells. Microfilaments were the only cytoplasmic components that could be detected within or in the vicinity of the penetrating pseudopods. No morphological signs of secretion could be observed in the contact region. Morphological aspects of the contact region are discussed as related to the lytic mechanism.     

Application of Enzyme Production Properties in Subtyping of Group A Streptococci According to T Type

The production of extracellular nicotinamide adenine dinucleotide glycohydrolase (NADG) and the cell-bound lipoproteinase (serum opacity reaction, SOR) by strains of different serological types of group A streptococci, in relation to the T typing, was studied. The production of both NADG and SOR, or only one of them, was found to be characteristic of serotypes, as determined by M and T antigen. No difference in the production of these enzymes was found in relation to M-positive and M-negative variants. Investigation into NADG and SOR production as related to the T type enabled the division of a single agglutination pattern into four main groups, each of which corresponds to one specific M type or more. Of the 370 strains belonging to 12 different T-agglutination patterns, 21% produced both enzymes and 42.5% failed to produce any of them, whereas the remaining 36.5% produced only one out of the two enzymes. Five streptococcal types which did not produce NADG and SOR also failed to synthesize streptolysin S at the early logarithmic phase of growth, indicating that streptolysin S production by young cultures may be also related to serotype. No correlation was found between the production of NADG-SOR as related to serotype and the production of streptolysin O, acid phosphotase, esterase, N-acetylglucosaminidase, hyaluronidase, streptokinase, and the cell-sensitizing factor. The practical and potential usefulness of NADG and SOR production in epidemiological studies is discussed.     

Covalent modification of the permeability pathways induced in the human erythrocyte membrane by the malarial parasite Plasmodium falciparum

The intraerythrocytic malarial parasite Plasmodium falciparum induces permeation pathways in the plasma membrane of its host, the red blood cell. The pathways display porelike properties with selectivity toward anions and neutral molecules. They are shown here to be susceptible to chemical modification by 4,4'-diisothiocyano-2,2'-dihydrostilbene disulfonic acid (H2DIDS), an amino-reactive reagent which is impermeant to uninfected cells. At pH 7.4 the reagent affected transport only marginally while freely entering into infected cells and reacting with intracellular hemoglobin. On the other hand, at pH above 8.5, the compound blocked the pathways efficiently (IC 50 approximately equal to 50 microM, at 37 degrees C for 10 min) as judged by four criteria: (1) selective lysis of infected erythrocytes in the presence of isotonic polyols; (2) uptake of [14C] sorbitol into infected cells; (3) uptake of the fluorescent anion Nbd-taurine into infected cells under conditions in which the native anion transport systems was inhibited; and (4) labeling of intracellular hemoglobin by the permeating reagent [3H]H2DIDS. The inhibitory effect was observed only with mature forms of parasitized cells, i.e., from the trophozoite stage and onward, while the pathways of immature ring forms were refractive. However, when the probe was incorporated into the interior of hemoglobin-depleted resealed ghosts prepared from ring forms, it was found to inhibit the pore-mediated transport. On the basis of these and other studies we postulate that the H2DIDS-sensitive sites on the pathways are endofacial, thus requiring penetration of the probe (probably through the same pathway) for their inactivation. Labeling studies with the radiolabeled modifier implicate 120-Kd, 63-Kd, and/or 51-Kd polypeptides as candidates for the pore components.     

Gangliosides indirectly inhibit the binding of laminin to sulfatides

Laminin, a glycoprotein of basement membranes, agglutinates aldehyde-fixed erythrocytes. Laminin-mediated hemagglutination is strongly inhibited by some gangliosides and anionic phospholipids. Laminin, however, binds only to sulfatides among the lipids extracted from erythrocytes. We now report that gangliosides are remarkably potent inhibitors of laminin binding to sulfatides when both lipids are adsorbed on plastic. A 50% inhibition of laminin binding to 100 ng of sulfatides is obtained with 10 ng of GM3 and 8 ng of GM1, respectively. Mixing of sulfatides with neutral glycolipids, phosphatidyl choline, or cholesterol does not inhibit laminin binding, whereas mixing with sulfatide-depleted erythrocyte lipids enhances binding. Inhibition of binding by gangliosides is not due to competition for adsorption to the plastic, as preincubation of the adsorbed lipids with neuraminidase reverses inhibition by GM3, but not by GM1 which is not a substrate for the enzyme. These results are consistent with the observations that treatment of fixed erythrocytes with neuraminidase increases their agglutinability by laminin and that pretreatment of erythrocytes with gangliosides followed by washing gives similar inhibition as seen when gangliosides are present as competitive inhibitors. Thus, inhibition of laminin-mediated agglutination by gangliosides probably results from masking of erythrocyte sulfatides due to adsorption of gangliosides onto the membrane rather than from a direct competition for laminin binding sites.     

Functional incorporation of the human erythrocyte chloride exchange system into plasma membranes of Friend erythroleukemic cells by Sendai virus-induced cell fusion

Human erythrocytes (RBC) were shown to exchange Cl⁻ by an exceptionally fast mechanism ( of ³⁶Cl⁻ equilibration at 1 °C is approx. 20 sec) which is demonstrably susceptible to specific inhibitors of anion exchange such as 4,4′-dinitrostilbene-2,2′-disulfonic acid (DNDS) and 4,4′-diisothyocyano-2,2′stilbene disulfonic acid (DIDS). Friend erythroleukemic cells (FELC) on the other hand, display both markedly slower Cl⁻ exchange rates ( of ³⁶Cl⁻ equilibration at 1 °C is approx. 60 min) and substantially lower susceptibilities to either DNDS or DIDS than RBC. After fusion between RBC and FELC, Cl⁻ exchange across FELC-RBC plasma membranes was noticeably enhanced compared with FELC. This enhancement was specificially abolished either by the addition of DNDS or by fusing FELC with DIDS-treated RBC.     

Analysis of plant root electropotentials

A model is proposed for the analysis of steady-state electropotentials in plant roots. The radial element of this model is discussed in terms of two parallel pathways, i.e. the symplasm and the extracellular space. The symplasm pathway is treated as a system of three compartments in series. The electrical potential difference across this pathway and its resistance are computed as a function of ion concentrations in the different compartments and the membrane properties. Similarly the extracellular pathway is analyzed as an ion-exchange membrane. Integration of both pathways into a parallel array analog and comparison with experimental data show that the widely used treatment of the root system as a single passive membrane is inadequate. It is also concluded that the occurrence of electrogenic pumps cannot be ascertained by straightforward inhibitor studies. However, the model proved to be insensitive to relatively wide variations in some of the parameters involved in the determination of its behavior.     

Evidence for active water transport in a corn root preparation

Summary Water flow was measured in aZea mays root preparation consisting of a segment from which the central part had been excised. It was shown that water flow had two components, one osmotic and one non-osmotic. The non-osmotic flow was inhibited by cyanide. No correlation was found between water flow and solute flow. These findings suggest that active water transport occurred in the root preparation. The mechanism of such water movement is discussed.