Effects of antibodies to choriogonadotropin in malignant growth

Summary We have studied the effects of preimmunization with a conjugate of the -subunit of choriogonadotropin and tetanus toxoid (CG-tt) on the growth of the implanted R 3230 AC mammary adenocarcinoma (in Fischer 344 rats) and the implanted 5123 1-1 hepatoma (in Buffalo rats) after 20 days, in order to determine if the in vivo production of antibodies against CG could modify the relationship between host and malignant growth. The results obtained demonstrated that active immunization against CG retarded significantly (P<0.01) the growth of the two transplantable tumors. Anti-CG antibodies were also determined and constantly found in the sera of all the preimmunized rats of both strains while no antibodies to CG were found in the control animals.     

STUDIES ON THE ANOMALOUS VISCOSITY AND FLOW-BIREFRINGENCE OF PROTEIN SOLUTIONS : III. CHANGES IN THESE PROPERTIES OF MYOSIN SOLUTIONS IN RELATION TO ADENOSINETRIPHOSPHATE AND MUSCULAR CONTRACTION

1. An investigation of the physicochemical properties of myosin has been carried out. Prepared under standard conditions, the ratio of flow-birefringence to protein concentration is uniform. The effect of electrolytes, pH, and urea on the flow-birefringence and viscosity (relative and anomalous) of myosin has been examined. 2. Decrease or abolition of flow-birefringence does not necessarily imply far reaching denaturation, since such effects can be reversed by a variety of means. 3. When a myosin solution is treated with adenosinetriphosphate, its flow-birefringence is decreased (average 48 per cent), its anomalous viscosity is retained, and its relative viscosity is decreased (average 14 per cent). The full effect of adenosinetriphosphate is obtained at 0.004 M; a molarity very much less than that of other substances which decrease the flow-birefringence of myosin. 4. The changes in the physicochemical properties of myosin brought about by adenosinetriphosphate are spontaneously reversible, and are connected with the enzymatic action of the protein as adenosinetriphosphatase. 5. Effects similar to those of adenosinetriphosphate on the physicochemical properties of purified myosin have been obtained so far only with inosinetriphosphate. 6. Inorganic phosphate is split off by myosin from inosinetriphosphate as well as from adenosinetriphosphate. Inorganic triphosphate is split by 1 to 2 per cent solution of three times precipitated myosin. 7. Adenosinediphosphate and inorganic triphosphate act as competitive inhibitors with adenosinetriphosphate, blocking the fall of flow-birefringence. 8. The implications of the results, and the conception of active enzymic groups attached to proteins participating in cell structure, whether contractile or non-contractile, are discussed in relation to present views on muscle physiology and other biological problems.     

The binding of phloridzin to the isolated luminal membrane of the renal proximal tubule

The binding of [³H]ploridzin by isolated luminal membranes of the rabbit proximal tubule and by slices of rabbit kidney cortex was studied.Kinetic analyses of the relationship between the concentration of phloridizin in the incubation medium and the binding of phloridzin to the membrane indicated two distinct classes of receptors sites. One class, comprising high affinity sites, reached saturation at 20–25 μM phloridzin, had a K(phloridzin) of 8 μM, and 8·10⁺² nmoles interacted with 1 mg of brush border protein. The other class, comprising low affinity sites, had a K(phloridzin) of 2.5 mM, and the number of binding sites was 1.25 nmoles/mg Na⁺ was required for the binding of phloridzin at the high affinity sites. Na⁺ decreased the apparent K_i for phloridzin; the apparent V of binding was not altered. Binding at the low affinity sites was independent of Na⁺. Ca²⁺ was necessary for maximal binding at the high affinity sites. Binding of phloridzin at high affinity sites was more sensitive to N-ethylmalcimide and mersalyl than was binding at low affinity sites. Binding at high affinity sites, but not at low affinity sites, was temperature dependent.d-Glucose was a competitive inhibitor of the high affinity binding of phloridzin. The apparent K₁ was 1 mM. D-Glucoe inhibited non-competitively at the low affinity sites. l-Glucose had no influence on phloridzin binding. Phloretin was a competitive inhibitor of high affinity phloridzin binding with an apparent K_i of 16 μM. Phloretin inhibited low affinity bindings of phloridizin non-competitively. Binding of phloridzin at high affinity sites was completely reversible. Binding at low affinity sites was only partially reversed. Phloridzin bound at high affinity sites on the brush border was displaced by phloridzin and phloretin but not by d-glucose.The mechanism of the high affinity binding of phloridzin was distinguished from that of the initial interaction of d-glucose with the membrane. Binding of phloridzin required Na⁺, whereas the interaction of d-glucose with the membranes had a prominent Na⁺-independent component.Intact renal cells in cortical slices accumulated phloridzin. The uptake did not saturate, was Na⁺ independent, and was not competitively inhibited by sugars. These characteristics resemble those for the low affinity binding of phloridzin by isolated membranes. It is suggested that low affinity binding may represent an initial binding followed by uptake of the glycoside into membrane vesicles.     

Studies on photophosphorylation utilizing methylene diphosphonate analogs of ADP and ATP

Spinach chloroplasts were able to photophosphorylate the ADP analog α,β-methylene adenosine 5′-diphosphate (AOPCP). Phosphorylation of AOPCP was catalyzed by chloroplasts that were washed or dialyzed to remove free endogenous nucleotides. In the presence of glucose, hexokinase, AOPCP and ³²P_i, the ³²P label was incorporated into α,β-methylene adenosine 5′-triphosphate (AOPCPOP).In contrast to photophosphorylation of AOPCP, the ATP analog AOPCPOP was a poor substrate for the ATP-P_i exchange reaction and its hydrolysis was neither stimulated by light and dithiothreitol nor inhibited by Dio-9.Photophosphorylation of AOPCP was inhibited by the α,β- and β,γ-substituted methylene analogs of ATP, while phosphorylation of ADP was unaffected by them. The ATP-P_i exchange was also unaffected by both ATP analogs, while the weak AOPCPOP-P_i exchange was inhibited by the β,γ-methylene analog of ATP.Direct interaction of methylene analogs with the chloroplast coupling factor ATPase was indicated by the enzymatic hydrolysis of AOPCPOP on polyacrylamide gels.     

Renal sugar transport in the winter flounder IV. Effect of CA2+ on sugar transport in teased renal tubules

The electrolyte distribution and sugar uptake by teased renal tubules of winter flounder (Pseudopleuronectes americanus) was studied using incubation media with 1.4 mM Ca²⁺ (controls) and without Ca²⁺. The omission of Ca²⁺: (a) produced some cellular swelling, and increase in cell Na⁺ and loss of K⁺; (b) had no effect on the extracellular propylene-glycol space; (c) increased the uptake of non-metabolizable methyl-α-D-glucoside by the tissue: whereas in controls the equilibrium tissue/medium ratio (T/M) for the Na⁺-independent uptake of the sugar was 0.71 ± 0.03 (SEM, n = 9), in Ca²⁺-free media the T/M rose in 1.18 ± 0.06. The increase in sugar uptake seen in absence of Ca²⁺ was abolished by absence of Na⁺ (Li⁺-media), 0.5 mM ouabain and 0.5 mM phlorizin; (d) produced an increase in the galactose phosphate level without affecting that of free sugar; (e) decreased the active uptake of 2-deoxy-D-galactose by the tubules. These results were analyzed on the basis of available information on the transport characteristics of the sugars at the luminal and antiluminal cell faces. It is suggested that absence of Ca²⁺ increases the permeability of the intercellular junctions, thus permitting sugars to enter from the external medium into the tubular lumen. Methyl-α-D-glucoside and D-galactose can then be taken up into the cells by brush-border localized active processes, whereas 2-deoxy-D-galactose is not reabsorbed at this cellular face. At 1.4 mM Ca²⁺, the intercellular junction appears to be relatively impermeable to sugars and the transport properties in teased tubules reflect then events predominantly localized at the antiluminal cell face.     

Effect of anisotonic media on cell volume and electrolyte fluxes in slices of the dogfish (Squalus acanthias) rectal gland

Summary Osmotic responses of slices of dogfish rectal gland to hypotonic (urea-free) and hypertonic media were studied. Transfer of tissue from isotonic (890 mosM) to hypotonic (550 mosM) saline produced an osmotic swelling associated with a slow net uptake of cell K⁺ (and Cl^–) and a slow, two-component efflux of urea. Media made hypertonic (1180 mosM) by addition of urea or mannitol produced osmotic shrinkage with a net loss of KCl. The cell osmotic responses in hypotonic media were lower than predicted for an ideal osmometer. No volume regulatory responses were seen subsequent to the initial osmotic effects. The cation influx in hypotonic media lacked specificity: in the presence of 0.5 mM ouabain or in K⁺-free media a net influx of Na⁺ was found. At steady state, the cell membrane potential evaluated from the Nernst potentials of K⁺ and triphenylmethyl phosphonium⁺, was independent of medium tonicity, suggesting the membrane potential as a determinant in the cellular osmotic response. Zero-time⁸⁶Rb⁺ fluxes were measured:⁸⁶Rb⁺ influx was not affected by hypotonicity, implying an unchanged operation of the Na⁺–K⁺-ATPase. On the other hand,⁸⁶Rb⁺ efflux was significantly reduced at hypotonicity; this effect was transient, the efflux returning to the control value once the new steady state of cell volume had been reached. A controlled efflux system is therefore involved in the cell osmotic response. The absence of the volume regulatory phenomenon suggests that the cells are not equipped with a volume-sensing mechanism.Abbreviations and symbols DW dry weight - E extracellular (polyethylene glycol) space - E Nernst potential - H₂O_e H₂O_i tissue water, extra- and intracellular - TPMP ⁺ triphenyl methyl phosphonium salt - WW wet weight     

A Bayesian analysis of multivariate doubly-interval-censored dental data

A Bayesian survival analysis is presented to examine the effect of fluoride-intake on the time to caries development of the permanent first molars in children between 7 and 12 years of age using a longitudinal study conducted in Flanders. Three problems needed to be addressed. Firstly, since the emergence time of a tooth and the time it experiences caries were recorded yearly, the time to caries is doubly interval censored. Secondly, due to the setup of the study, many emergence times were left-censored. Thirdly, events on teeth of the same child are dependent. Our Bayesian analysis is a modified version of the intensity model of Harkanen et al. (2000, Scandinavian Journal of Statistics 27, 577-588). To tackle the problem of the large number of left-censored observations a similar Finnish data set was introduced. Our analysis shows no convincing effect of fluoride-intake on caries development.