Mode of reconstitution of chicken erythrocyte and reticulocyte chromatin. The mode of reconstitution of chicken erythrocyte and reticulocyte chromatin has been investigated. Chromatin was dissociated in 2 M NaCl, 5 M urea, and 0.01 M potassium phosphate (pH 7.2) and was dialyzed against various NaCl concentrations in 5 M urea and 0.01 M potassium phosphate (pH 7.2). Histone reassociation to DNA occurs with the binding of histone H5 at 0.5 M NaCl in 5 M urea, followed by histone H1 at 0.4 M NaCl in 5 M urea. All the classes of histones are reassociated with DNA at 0.2 M NaCl in 5 M urea and binding of all classes of histones is complete in 0.1 M NaCl and 5 M urea. Nonhistone proteins reassociate with DNA before and at the same time that histones reassociate with DNA. Binding of nonhistone proteins to DNA appears to be complete in 5 M urea and 0.01 M potassium phosphate (pH 7.2). There is also found in both erythrocyte and reticulocyte chromatin a nonhistone protein present in relatively high concentrations, which remains associated with DNA in 2 M NaCl and 5 M urea. This tightly bound protein appears as one major band when chromatographed on sodium dodecyl sulfate-polyacrylamide gels, with a molecular weight of 95 000. This protein is soluble in phenol and sodium dodecyl sulfate but is insoluble in 5 M urea or 4 M guanidine hydrochloride. A fraction of reticulocyte nonhistone proteins was found to bind to DNA-cellulose in 5 M urea. The majority of these proteins elute at 0.15 M NaCl in 5 M urea but a significant fraction elutes at NaCl concentrations at which the bulk of the histones do not bind to DNA. The proteins that bind to free DNA have low molecular weights and do not show species speciificity. Approximatley 50% of the reticulocyte nonhistone protein does not bind to a DNA-cellulose column in 5 M urea and may require histones for complete reassociation.


Comparison of the phosphorus magnetic resonance and circular dichroism properties of calf thymus DNA and chromatin. Dual measurements of the 31P magnetic resonance spectra and the circular dichroism spectra have been made on calf thymus DNA and purified chromatin. The 31P magnetic resonance signals for all samples fell at 1.2 +/- 0.1 ppm relative to 85% orthophosphoric acid. The full width at half-height of the signal of samples in which the molecular weight of the native DNA component was in the 2-9 X 10(6) range was ca. 50 Hz. This bandwidth was reduced dramatically to ca. 20 Hz by reducing the molecular weight to 140 000 (by sonication) or by heat denaturation of the high-molecular-weight DNA. The position of the signal and the bandwidth of the chromatin samples did not differ significantly from that of the DNA samples of comparable molecular weight and state of nativity. The intensities of the chromatin signals, however, were all less than those of signals of DNA in companion runs conducted under comparable experimental conditions. The reduction of the intensity of the magnetic resonance signal paralleled the lowering of the intensity of the positive band above 260 nm in the circular dichroism spectrum of the given sample of chromatin relative to the spectrum of protein-free DNA. In fact, the percent reduction of the magnetic resonance signal of chromatin relative to protein-free DNA was, within experimental error, equal to the percentage of nucleotide residues in the Watson-Crick B secondary structure. Since the latter fraction of residues can be correlated with those in the interbead regions of the superstructure of chromatin, we have concluded that the signal of the nucleotide residues in the beads, or v bodies, has been broadened to the point of extinction by a packing arrangement which maximizes phosphate-protein interactions and structural rigidity.


Near-ultraviolet tyrosyl circular dichroism of pig insulin monomers, dimers, and hexamers. Dipole-dipole coupling calculations in the monopole approximation. The tyrosyl circular dichroism (CD) has been calculated using the conformation of pig insulin observed in rhombohedral crystals containing 2 zinc atoms per hexamer. These calculations predict that the tyrosyl CD at 275 nm will be enhanced disproportionally as monomers interact to form dimers and as dimers interact to form hexamers. This enhanced tyrosyl CD (delta epsilon per 5800 molecular weight) results from new coupling interactions generated in the regions of contact between monomers and between dimers. These calculations illustrate that a large CD enhancement may accompany aggregation even in the absence of a conformation change in eith monomer. The tyrosyl CD intensities calculated for monomers, dimers, and hexamers of 2-zinc pig insulin are compatible with the experimentally observed CD spectra which are enhanced about fourfold in the hexamer compared with the monomer. Zinc ions and other metals do not contribute directly to the tyrosyl CD but only influence the optical properties by promoting the hexameric state. The relation of the integrity of the molecule to dimer formation and the biological activity of the molecules are discussed. The largest calculated contributions to tryosyl CD arise from interactions with far-ultraviolet transitions of neighboring aromatic groups. In the hexamer, about half of the tyrosyl CD intensity is calculated to arise from Tyr-A14.


An investigation of the electronic and steric environments of tyrosyl residues in ribonuclease A and Erwinia carotovora L-asparaginase through fluorescence quenching by caesium, iodide and phosphate ions. The fluorescence lifetimes and relative quantum yields of several derivatives of tyrosine are reported. The quenching of the fluorescence of these compounds by phosphate, caesium and iodide ions has been investigated; the encounter rate constants, calculated from the quenching parameters and lifetimes, show a clear dependence on the charges borne by the quenchers and fluorophores. The ratio of the Stern-Volmer constants of iodide and caesium, ions of similar size, defines an electrostatic parameter sensitive to the charge of the fluorophore which can be evaluated without knowledge of the fluorescent lifetimes. The mean of the encounter rate constants for caesium and iodide ions defines a rate constant which is largely charge-independent and is used to establish a steric parameter. The two parameters are used to investigate the tyrosine environment in bovine ribonuclease A (EC 3.1.4.23) and Erwinia carotovora L-asparaginase (EC 3.5.1.1). The quantum yield of L-asparaginase (0.12) is very high for a class A protein and may be associated with the absence of disulphide bridges. There was no evidence for more than one type of tyrosine residue from the quenching experiments with either enzyme, an observation which is attributed to efficient energy transfer amongst tyrosine residues. At pH values close to the isoelectric points of the enzymes the electrostatic parameter suggests that the environment of the quenchable tyrosines in L-asparaginase is somewhat more positive than in ribonuclease. In 1% sodium dodecyl sulphate the tyrosine environment of L-asparaginase becomes markedly negative as expected. The steric parameter indicates a lower accessibility of the tyrosine residues in L-asparaginase than in ribonuclease; an illustrative calculation is provided linking the steric parameter with the number of exposed tyrosine residues by taking into account the greater collision frequency of the larger protein molecules and the encounter distance for quenching determined from charge effects on the quenching of the model compounds. The calculation suggests that three tyrosyl residues are accessible in ribonuclease, in good agreement with other studies, but in L-asparaginase the number increases from 0.4 at pH 5.73 to 0.8 at pH 9.16 suggesting a loosening of the enzyme structure at high pH.


Biosynthesis of mammalian transfer RNA. Evidence for regulation by deacylated transfer RNA. The rate of tRNA synthesis in cultured Friend leukemia cells has been examined as a function of the variation in polyribosome structure produced by treatment with a variety of inhibitors of protein synthesis. The results indicate, in contrast to the conclusions of Bölcsföldi (Bölcsföldi, G. (1974) Exp. Cell Res., 88, 231--240), that no necessary relationship exists between the ribosome distribution and the rate of tRNA synthesis. Alternatively, it is observed that inhibitors of tRNA aminoacylation cause, in all cases, a decrease in the rate of tRNA synthesis whereas drugs which may stimulate the aminoacylation of tRNA cause, in all cases, an elevation of the rate of tRNA synthesis. It is concluded that tRNA synthesis in mammalian cells may be regulated by the relative levels of acylated and deacylated tRNA.


The adsorption of lysine vasopressin at ionized interfaces. It has been shown by surface potential measurements that lysine vasopressin and oxytocin may be bound by ionic surfaces to very varied extents. To dodecyl sulphate and phosphatidylserine monolayers the binding is very strong and is comparable to that for biological receptors such as those in toad bladder. For dioleyl phosphate and the carboxyl group of the polypeptide alamethicin, the binding is rather weaker while, for the zwitterionic lipids phosphatidylcholine and phosphatidylethanolamine, and for the erythrocyte surface, which contains two varieties of carboxylic acid group, no interaction seems to take place. In no system does the lysyl amino group of the vasopressin appear necessary for adsorption and, in the dodecyl sulphate monolayers, the interaction is strong even when the ionization of the terminal alpha-amino group is suppressed.


Cultured cells from renal cortex of hibernators and nonhibernators. Regulation of cell K+ at low temperature. Cells were grown as primary monolayer cultures from kidney cortex of guinea pigs (nonhibernators), hamsters and ground squirrels (both hibernating species). When plates of cells were placed at 5 degrees C, cells of guinea pigs lost 37% of their K+ in 2 h and those of the hibernator lost about 10%. Uptake of 42K into the cells exhibited a simple, single exponential time course at both temperatures. Unidirectional efflux of K+ was equal to K+ influx in all cultures at 37 degrees C and, within limits of error, in hibernator cells at 5 degrees C. Efflux was 3-to 5-fold greater than influx in guinea pig cells at 5 degrees C. After 2 h in the cold the ouabain sensitive K+ influx remaining (7-15% of that at 37 degrees C) was about the same in the cells of the 3 species. Cells from active hamsters and from hibernating ground squirrels, however, exhibited significantly greater pump activity after 45 min in the cold (19 and 14%, respectively). The stimulation of K+ influx by increasing [K+] did not show an increase in Km+ at 5 degrees C in cells of guinea pigs and ground squirrels. Lowering [K+]c and/or raising [Na+]c by treatment in low- and high-K+ media caused only slight stimulation of K+ influx, except in cells of ground squirrels at 5 degrees C in which the stimulation was at least 11-times greater than at 37 degrees C or in cells of guinea pigs at either temperature. This altered kinetic response of K+ transport to cytoplasmic ion stimulation with cooling accounted for about one-third of the improved regulation of K+ at 5 degrees C in ground squirrel cells; the other two-thirds was attributable to a greater decrease in K+ leak with cooling. The inhibition of active transport by cold in all 3 species was much less severe than that previously seen in any (Na++K+)-ATPase of mammalian cells.


Subcellular distribution of DNA-binding proteins from cultured hamster fibroblasts. The distribution between nuclei and cytoplasm of DNA-binding proteins from growing NIL cells was studied. To obtain the subcellular fractions, cell monolayers or cells previously detached from the culture dish were treated with non-ionic detergent N onidet P-40. Proteins with affinity for DNA were isolated from nuclear or cytoplasmic fractions by chromatography on DNA-cellulose columns and were further analyzed by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The results show that P8, one of the major components in the 0.15 M NaC1-eluted proteins, is found predominantly in the cytoplasmic fractions, whereas P6, the other main protein peak in this eluate, is more prominent in the nuclear fraction. Among the other proteins eluted at 0.15 M NaC1 from the DNA-cellulose column, P5 and P5' are detected in both nuclear and cytoplasmic fractions. All the other proteins in the 0.15 M NaC1 eluate are present almost exclusively in the cytoplasmic fraction. On the other hand, most of the proteins with higher affinity for DNA, eluted from the column at 2 M NaC1, are present in the nuclear fraction, although they are also detected in the cytoplasm in amounts similar to those observed in the nuclei.


Comparative studies of human and chicken retinol-binding proteins and prealbumins. Microheterogeneity of retinol-binding proteins of human plasma and urine, and of chicken plasma was studied by polyacrylamide gel electrophoresis. All three protein systems were found microheterogenous. Incorporation of retinol into the protein preparations on the one hand, and depletion of these proteins from retinol on the other hand, enabled us to clarify the extent to which the presence or absence of the ligand affects the apparent heterogeneity. Upon electrophoresis, each of the native proteins displayed two pairs of protein zones. It appeared that within each pair the fast moving band corresponded to aporetinol-binding protein which upon binding of retinol was converted to a holoprotein with a slightly lower mobility. However, it did not seem that proteins of one pair were converted to proteins of the second pair upon binding of retinol, substantiating ghe microheterogenous character of this protein system. A rapid, two step procedure for isolation of prealbumins from plasma is described. The method which consists of DEAE-cellulose chromatography follwed by preparative electrophoresis was utilized to separate human and chicken prealbumins. Routine dodecyl sulphate electrophoresis resulted in partial dissociation of human prealbumin but in no dissociation of the chicken protein. More drastic treatments prior to electrophoresis were needed to effect complete disruption of both proteins into subunits.


The interaction between retinol-binding proteins and prealbumins studied by fluorescence polarization. The interaction between retinol-binding proteins and prealbumins of human and chicken was studied by fluorescence polarization techniques. The binding affinity between chicken plasma retinol-binding protein and chicken prealbumin was essentially the same as between the respective human proteins. Human urine retinol-binding protein displayed a similar affinity, though possibly slightly smaller than that of the human plasma protein, toward human prealbumin. Retinol-binding proteins and prealbumins of human and chicken have been found to cross-interact displaying an affinity similar to that displayed by the proteins of the same species. Solution of a binding equation which assumes identical, independent sites, indicated that the number of binding sites on prealbumin for retinol-binding protein is somewhat less than 2 with the human system, and in the neighborhood of 4 with the chicken system. A possible interpretation suggests that prealbumin possesses four identical binding sites for retinol-binding protein, one for each subunit, but that the binding is of a negative cooperative nature. A major share of the negative cooperativity is likely to result from steric hindrance induced by already bound retinol-binding protein molecules, which have a sizable volume compared to the volume of the prealbumin molecule. The cooperativity is likely to be more pronounced with the human system. Rotational relaxation times derived from Perrin plots suggest that 1:1 molecular complexes of retinol-binding proteins with prealbumins have a compact structure.


Fluorescence and the structure of proteins. XXII. Fluorescence of aminotyrosyl residues formed in ribonuclease A. 1. Tyrosyl residues on ribonuclease A were nitrated with tetranitromethane and then reduced to aminotyrosyl residues. By variation of reaction conditions and degree of exposure of tyrosyl residues it was possible to convert from 1 to all 6 tyrosyl to aminotyrosyl residues. 2. At the lower levels of 1-3 aminated tyrosyl residues/molecule the change in conformation seemed minor and 70% of the enzymatic activity was retained. When the three buried tyrosyl residues or all six residues were aminated only 5% of the enzymatic activity was retained. 3. Titration data, susceptibility to urea denaturation, and fluorescence characteristics indicated that some of the aminotyrosyl residues were buried in the interior and others were exposed on the surface of the protein. On the basis of the activation/emission wavelengths it was possible to distinguish buried (288/320 nm) and exposed (288/365-395 nm) aminotyrosyl residues as well as exposed tyrosyl residues (275-305 nm). 4. The modification of specific tyrosyl residues on a protein to aminotyrosyl residues appears to have some promise for observation of changes in environment of the residues that accompany various conformation changes by monitoring the fluorescence.


Acyl-CoA synthetases in guinea-pig liver mitochondria. Purification and characterization of a distinct propionyl-CoA synthetase. Guinea-pig liver mitochondria contain three soluble ATP-dependent acyl-CoA synthetases: (a) a medium-chain acyl-CoA synthetase, (b) a salicylate activating enzyme, and (c) a propionyl-CoA synthetase. A complete separation of these enzymes has been accomplished and the resulting preparation of propionyl-CoA synthetase (Spec. act. 4 units/mg protein) accepts acetate, propionate and butyrate as substrates with a high preference for propionate.


Anti-L serum. Two populations of antibodies affecting cation transport in LK erythrocytes of sheep and goats. Isoimmune sheep anti-L serum was fractionated, yielding two antibodies with different specificities of action on potassium transport in LK red cells of sheep and goats: anti-Lp, which stimulates active transport, and anti-L1, which inhibits passive transport.


1-O-acyl derivatives of glucose as non-penetrating inhibitors of glucose transport by hamster small intestine in vitro. 1-O-acyl derivatives of glucose are not transported by the hamster small intestine in vitro. These derivatives, however, are potent inhibitors of the glucose transport system. 1-O-Decanoyl glucose is a competitive inhibitor of beta-methyl glucoside transport.


Interaction of short chain and long chain fatty acid phosphoglycerides and bile salts with prothrombin. An equimolar mixture of phosphatidylserine and (dioleoyl) phosphatidyl-ethanolamine could substitute for brain cephalin preparations in the single stage prothrombin assay. However, no clot promoting activity was observed on the addition of any of the individual long chain fatty acid-containing phospholipids. Short chain fatty acid-containing phospholipids, such as diheptanoylphosphatidylcholine, diheptanoylphosphatidylethanolamine, diheptanoylphosphatidic acid, and dihexanoylphosphatidylcholine, or dihexanoylphosphatidylethanolamine were inhibitory under all conditions studied. Similar effects of these two general classes of phospholipids were observed in a two-stage thrombin generation system, in which a mixture of bovine Factor Xa, Factor Va, and Ca2+ were interacted with prothrombin. In the presence of 25 mM Ca2+, dioleoylphosphatidic acid or brain phosphatidylserine alone, and with other long chain phospholipids, formed complexes with bovine plasma prothrombin. On the other hand, dioleoyl-, diheptanoyl- or dihexanoylphosphatidylcholine under comparable conditions showed no binding to prothrombin. There appeared to be a small degree of binding of diheptanoylphosphatidic acid to prothrombin, but it was insufficient to cause any significant change in apparent molecular weight of prothrombin. A mixture of prothrombin, Factor V, diheptanoylphosphatidic acid/diheptanoylphosphatidylcholine and Ca2+ eluted in the void volume of Sephadex G-200, but showed a much reduced coagulant activity. Though a net negative charge on the phospholipid surface is required for phospholipid-protein interactions, this does not necessarily promote coagulant activity. Bile acids and bile salts, such as cholic acid, deoxycholic acid, taurocholic acid, glycocholic acid, lithocholic acid and dehydrocholic acid, exerted varying levels of stimulation on the prothrombin assay and thrombin generation system, but were not as effective as the phospholipids. Interestingly, no interaction of these bile acids or salts with prothrombin was noted in the presence of Ca2+. The results of these experiments suggest that negatively charged micelles per se are not sufficient for binding alone and that other chemical and physical characteristics of phospholipids are of prime importance.


Interactions of the components of the prothrombinase complex. The interaction of components of the prothrombinase complex, i.e. bovine Factor X or Factor Xa, bovine Factor V or Factor Va, phospholipid, and Ca2+, in various combinations was studied primarily by a gel filtration technique. In experiments, in which phospholipids ranging from those isolated from naturally occurring sources to those long chain (18 : 1) as well as short chain 6 : 0 and 7 : 0 fatty acids prepared by chemical and enzymatic synthesis were used, it was evident that a net negative surface charge on the lipid dispersions was one of the important requirements for interaction. Though the short chain fatty acid phospholipids interacted with the proteins of the prothrombinase complex, there was invariably a diminution in the activity of the enzyme complex. It was established that Factor V or Va did not bind Ca2+ and that the binding of either of these factors with phospholipids (with a net negative charge) was not dependent on Ca2+. However, the interaction of Factor X or Factor Xa with phospholipids with a negative charge required Ca2+. It was shown that Factor X could bind to the same type of lipid of lipid surface as that noted for Factor Xa. Of interest was the apparent difference in the phospholipid binding characteristics of the two variant forms of bovine plasma Factor X, i.e. X1 and X2, which might in part explain the differences in their specific activities. Of importance was the lack of demonstrable complex formation between Factors II, X and V in the absence of phospholipids and/or in the presence or absence of Ca2+. The significance of these results as they might apply to the configuration of the prothrombinase complex and its interaction with prothrombin plus  the usefulness of the short chain fatty acid phospholipid in exploring these lipid-protein interactions are discussed.


Galactosyltransferase activities during embryonic development of chick neural tissue. Galactosyltransferase specific activities in embryonic chick retina, optic tectum, and telencephalon were found to decline during embryonic development. Incorporation of galactose from nucleotide sugar into exogenously added glycoprotein acceptor was measured in the presence of excess of glycoprotein acceptor. This ensured that the specific activity measurements were reflections of tru enzyme specific activity rather than availability of acceptor. Moreover, we have shown that the decline in specific activity is not due to degradation of the nucleotide sugar, UDP-galactose, under our in vitro assay conditions. Enzymatic specific activity declined sharply with embryonic age for all tissues tested. This decline was not affected by the presence of 5-bromodeoxyuridine during in vitro culture of embryonic chick neural retina above that caused by the culturing alone. Galactosyltransferase activity was not found to be associated with the plasma membrane fraction from homogenized tissue but rather with the microsomal fraction. Thus, the changes in galactosyltransferase specific activity detected here do not reflect changes at the cell surface.


Lactoperoxidase-catalyzed lipid peroxidation of microsomal and artificial membranes. Lactoperoxidase, in the presence of H2O2, I-, and rat liver microsomes, will peroxidize membrane lipids, as evidenced by malondialdehyde formation. Fe3+ assists in the formation of malondialdehyde. Fe3+ can be added at the end of the reaction period as well as at the beginning with equal effectiveness, suggesting that it only acts to assist in the conversion of lipid peroxides, previously formed by lactoperoxidase, to malondialdehyde. The addition of EDTA to the microsomal reaction mixture results in a 40% decrease in malondialdehyde formation. The antioxidant butylated hydroxytoluene will completely block the formation of malondialdehyde. Malondialdehyde formation is not dependent upon the production of superoxide, singlet oxygen, or hydroxyl radicals. Peroxidation of membrane lipids by this system is equally effective in both intact microsomes and in liposomes, indicating that iodination of microsomal protein is not required for lipid peroxidation to occur.


Approximately 70% of fucose-labeled glycopeptides from the cell surface and cellular material of rat. Approximately 70% of fucose-labeled glycopeptides from the cell surface and cellular material of rat fibroblasts (3Y1B cells) were hydrolyzed by endo-beta-N-acetylglucosaminidase D in the presence of neuraminidase, beta-galactosidase and beta-N-acetylglucosaminidase. Structure of the susceptible glycopeptides were found to be very similar to non-membrane glycopeptides of the complex heteropolysaccharide unit, such as the sialylated glycopeptides of thyroglobulin. On the other hand, the resistant glycopeptides were also refractory toward endo-beta-N-acetylglucosaminidase H and alpha-mannosidase, and appeared to be a mixture of glycopeptides with unique structures.


Reaction of rhodanese with dithiothreitol. The reaction between bovine rhodanese (thiosulfate:cyanide sulfurtransferase, EC 2.8.1.1) and reduced dithiothreitol has been studied. This reagent, in the absence of thiosulfate, reduces the amount of sulfur carried by rhodanese with formation of sulfide and oxidized dithiothreitol: E-S-SH + reduced dithiothreitol replaced by E-SH + HS- + oxidized dithiothreitol, (E = enzyme). An inactivation was observed at high dithiothreitol/enzyme ratios or at very low enzyme concentrations. The inactivation was not observed in the presence of thiosulfate and can be reversed by cyanide or thiosulfate. A thiosulfate reduction activity of rhodanese was also found using dithiothreitol as reductant.


Chemotactic activity from rabbit peritoneal neutrophils. Lack of identity with N-acetyl-DL-phenylalanine beta-napthyl esterase. The chemotactic and N-acetyl-DL-phenylalanine beta-naphthyl esterase activities of rabbit peritoneal neutrophils are separable from each other by both DEAE cellulose and Sephadex G-100 column chromatography. Partially purified esterase obtained from DEAE-cellulose chromatography had molecular weight of 70 000. However, the partially purified fraction contained chemotactic activities with major activity in molecular weight of 28000 and minor activities in the molecular weights of 45000, 21900, 14500 and 10500. Esterase activity is inhibited by 10(-7) M p-nitrophenylethyl-5-chloropentylphosphonate but chemotactic activity is not.


Postnatal effects of chemically induced intrauterine growth retardation on some hematological values in the rat. The effects of artificially induced intrauterine growth retardation on hematological values of the rat are manifest only temporarily in the red blood cell number, the amount of hemoglobin, and the 2,3-DPG. These three elements reached normal levels after the fifth day of postnatal life. We believe that this temporary effect may be the consequence of the short-term effect of hydroxyurea (the drug used to produce intrauterine growth retardation) upon DNA synthesis in the hematopoietic system. The animals compensate for the low red cell number and low hemoglobin by raising the levels of 2,3-DPG brought about by the temporary anoxia.


Postnatal decline of maternally acquired antibodies in the rat. The decline (half-life) of maternally acquired antibodies of two different specificities (diphtheria and tetanus) was studied in rats. Essentially similar half-lives for antibody of each specificity were found in young rats from the same litter. In contrast, half-lives varied from 2 to 10 days in individuals of different litters. Significant differences in half-lives were demonstrated in young rats possessing markedly different initial titres of antibodies of the two specificities. It was established that there was a relationship between the half-life and the initial titre of antibody of each specificity which could be described by a rectangular hyperbola. These findings are in close aggreement with previous studies in human infants.


An application of the clumped binomial model to the analysis of clustered attribute data. This paper is concerned with the statistical analysis of situations where the experimental units are clusters of subjects, each of whom is classified with respect to the presence of a particular (binary response) attribute. For this purpose, the clumped binomial probability model is used to define an underlying mean rate parameter in terms of which the effects of certain treatments as well as cluster size itself can be investigated. The resulting methodology involves a synthesis of certain maximum likelihood and weighted least squares procedures; it is illustrated for a three-dimensional contingency table pertaining to numbers of deaths in litters of baby mice.


Lymphocytopoiesis in vitro. Production of small lymphocytes and plasma cells in the isolated perfused spleen. The production of small lymphocytes and mature plasma cells could be shown in the isolated perfused pig spleen using labelling with 3H-TdR. Within the first day the labelling index for lymphocytes increased to up to 10% and the labelling index for mature plasma cells to up to 60%.


Anti-Müllerian hormone: estimation of molecular weight by gel filtration. Incubation medium of calf fetal testes was submitted to gel filtration on Sephades G 200 and Bio-Gel A-5 M, IN ORDER TO DETERMINE THE MOLECULAR WEIGHT OF THE ANTIMULLERIAN HORmone (A.M.H.) RESPONSIBLE FOR THE REGRESSION OF MULLERIAN DUCTS IN MALE FETUSES. Results from both series of experiments indicate a molecular weight in the range between 200,000 and 295,000 daltons, even in the presence of high concentration of a reducing agent.


The propulsive mechanism of the dinoflagellate transverse flagellum reconsidered. As a result of the recent discovery that the transverse flagellum of dinoflagellates does not beat helically within the cingulum it has been necessary to formulate a new model for the manner in which this flagellum can impart a forward motion to the cell. It is theorised that the contribution to forward propulsion by the transverse flagellum can only arise directly from an asymmetry of the flagellar wave. In addition we have drawn attention to topographic features of the cell, and particularly of the left sulcal list, which may augment this thrust by redirecting cingular flow down the sulcus.


[Role of interoceptive signalization in modulating emotional-behavioral responses]. Chronic experiments were conducted on rabbits; electrodes were implanted into the deep structures of the brain. Stimulation of gastric receptors led to modulation of the emotional-behavioristic reactions induced by electric stimulation of the hypothalamus, the amygdala and the hypocampus. The effect depended on the intensity of interoceptive stimulation and peculiarities of the emotional reaction with own cerebral control systems.


[Facilitation of the effects of L-DOPA by alpha-methyldopa]. Alpha-methyl-DOPA increased the duration of excitation induced by L-DOPA in mice, alpha-methyl-DOPA potentiated the anticataleptogenic activity of L-DOPA in mice pretreated with reserpine or haloperidol. Alpha-methyl-DOPA given to cats 30 min prior to L-DOPA diminished the influence of L-DOPA on the blood pressure and the contraction of the nictitating membrane. When given 4 to 6 hours prior to L-DOPA alpha-methyl DOPA enhanced the reaction of the blood pressure and contraction of the nictitating membrane induced by L-DOPA or dopamine.


[Mechanism of antibody formation suppression in high density suspension cultures]. There was a sharp decrease in the number of antibody producing cells formed in induction of primary immune reaction in vitro with increase of surface density (number of cells per 1 cm2 of the bottom of the incubation vessel); the cause of this phenomenon was studied. It was shown that depression of antibody formation in dense cultures was associated not with disturbance of the induction of the clone of antibody-forming cells, but with the inhibition of the subsequent proliferation of the cells of this clone. Disturbance of proliferation was reversible: a reduction of the density of the cultures 1 or 3 days after the beginning of the incubation led to the restoration of the growth of the number of antibody-producing cells. The depressive effect could not be transferred by humoral way from the cultures with a high density to the cultures with a low density; it could not be eliminated by daily replacement of the incubation medium in dense cultures for a fresh nutrient medium.


[Metastasis of sarcoma 45 and 536 following different methods of inoculation]. Intertesticular inoculation to rats of 1 ml of sarcoma suspension was accompanied by a rapid growth of the tumour at the site of inoculation and the spread of the tumour nodes to the omentum majus, porta hepatis, mesentery and the abdominal walls. After intraperitoneal inoculation the spread of the tumour nodes was similar. In these experiments animals died in 13-15 days. One month after the intratesticular inoculation of 0.2 ml of the tumour suspension-lung metastases were revealed in 74% of the animals with sarcoma-45 and in 90% of those with sarcoma-536. It was shown that tumour cells appeared in the blood stream already during the inoculation. However, laying of lung metastases occurred only after the formation of a primary tumour node at the site of transplantation. A conclusion was drawn that this system was a suitable experimental model for the study of the lung metastases development in rats.


[Quantitative immunochemical differentiation of creatine kinase isoenzymes. Use of antiserum to isoenzyme MM]. A possibility of quantitative differentiation of human isoenzymes of creatine kinase by using the antiserum to isoenzyme MM was studied. Rabbit antiserum to human isoenzyme MM proved to specifically inhibit its activity, without altering the isoenzyme BB activity. The use of antiserum to isoenzyme MM as a specific inhibitor makes possible differential determination of the isoenzymes of creatine-kinase or their subunits in the artificial isoenzyme mixtures, and also in the human blood serum and human tissue homogenates.


[Interaction of 17beta-estradiol with histones. Effect of the hormone on partially reconstructed mucleohistones]. Binding of 17beta-estradiol with histones of calf thymus and the influence of the hormone on heart denaturing of partially reconstructed nucleohistones were investigated. By the degree of the hormone binding histone fractions could be arranged in the following order: F2a greater than F3 greater than F1  greater than or equal F2b. Addition of 17beta-estradiol gave rise to increase of the first and a corresponding decrease of the second melting phase of partially reconstructed nucleohistones F2a, F3 and F2b. Addition of the hormone to the partially reconstructed nucleohistone F1 gave rise to the opposite changes in the heat denaturing profile.


[Role of different types of cells in the effect of immunoglobulin synthesis stimulation in mixed cultures]. The radioisotopic method with the use of immunosorbents was applied to the study of the synthesis of antibodies and nonspecific immunoglobulins in the mono- and mixed cultures of cells of the immune lymph nodes and intact bone marrow in removal of the adherent or T-cells. Treatment of the lymph node cell population with anti-O-serum decreased the antibody synthesis to 30%, whereas removal of the adherent cells - to 70%. In combined cultivation of the cells of immune lymph nodes, devoid of adherent cells, with the intact bone marrow cells the effect of stimulation of immunoglobulin synthesis was not eliminated, whereas treatment of cells of the immune lymph nodes with anti-O-serum doubled this effect. Possible mechanisms of the effect of stimulation of the immunoglobulin synthesis in mixed culture is discussed, and a conclusion was drawn on the necessity of cooperation between the individual types of cells during the productive phase of the immune response.


[Changes in the number of stem cells during mouse bone marrow cultivation on a sublayer of fibroblast-like cells]. A determination was made of the number of colonies in the spleen of irradiated mice after administration to them of a culture of mouse bone marrow cells. Colony-forming units proved to persist in the culture only for a short time. Use of preliminarily grown underlayer of fibroblasts of bone marrow origin had no effect on the perservation and dynamics of the changes in the number oc colony-forming units.


[Cytogenetic characteristics of lung tumors induced by exposing BALB/C mice to transplacental N-nitrosoethylurea]. Chromosome pattern of lung adenocarcinomas in BALB/c mice exposed to N-nitrosoethylurea during prenatal life (transplacental action) was studied; it appeared that 121 of 145 metaphases examined contained 40 chromosomes. Along with this, there were revealed individual hypo-, hyperdiploid and tetraploid cells. Chromosomes possessed the usual telecentric structure in all the cells. The problem of possible changes of kariotypes of the tumours under study indetectable by the routine staining techniques is discussed.


[Concentration of monoamines and activity of several enzymes in the arcuate nucleus of the hypothalamus in young and aging rats during the estrous cycle]. The arcuate nucleus (AN) and the median eminence (ME) of the hypothalamus were investigated in young and ageing female rats. During the estral cycle (EC) the monoamine (MA) content, the monoaminoxidase (MAO), NADP and NAD-diaphorase activities were determined in the AN, and the MA content and the activity of alkaline phosphatase (AP) -- in the ME. In young rats in the proestrus-estrus there was an increase in the activity of the NADP and NAD-diaphorase and of the MA content, but a decrease of the MAO activity. This indicated an intensified function of the nucleus at these stages of the EC. Accumulation of the MA in the ME was noted in the diestrus, while in the proestrus their concentration sharply fell; on the other hand, the activity of the AP was considerably increased. In the ageing rats the dynamics of the indices under study during the EC were largely unchanged. However, the functional activity of the AN proved to increase, and in the ME and elevation of the MA concentration and disturbance of its release from the nerve terminals was seen.


[Disorders in the development of offspring following exposure of rats to hydrogen chloride]. Female Wistar rats were subjected to the influence of hydrogen chloride before and on the ninth day of pregnancy. Apart from marked lung lesions there were functional disturbances in the kidneys and the liver in both groups of the experimental animals. Changes in the state of maternal organs caused changes in the development of organs in the progeny (functional derangement of these organs during the postnatal period). Disturbances in the lungs (after an additional load) and in the kidneys were noted in the male progeny of both groups; there were also disturbances of the hepatic function in the male progeny when their mothers were acted upon by HC1 before the occurrence of pregnancy.


Relationship between childhood infections and measured intelligence. The possibility that some of the common childhood infections lead to unrecognized impairments of neurological function was examined in 43 820 Birmingham children whose intelligence was assessed in the 11-plus examination. Mean verbal reasoning scores were lower for children who had had measles or pertussis than for those who had had neither of these diseases. However, since attack rates and measured intelligence are related inversely to social class, the lower scores of children with measles and pertussis may be due to class differences which are not eliminated completely by standardization for maternal age and birth order. Mean scores were a little higher for children who had had rubella than for those who had not, and it is suggested that this difference may be due to more frequent reporting of the disease by the more intelligent mothers.


Follow-up study of small-for-dates babies. A group of small-for-dates full-term babies whose intra-uterine growth was followed by serial ultrasonic cephalometry were examined at a mean age of 4 years. Those children whose skull growth had begun to slow in utero before 34 weeks' menstrual age were more likely to have a height and weight less than the 10th centile. When the onset of growth failure had occurred before 26 weeks there was a lower developmental quotient at follow-up using the Griffiths extended scales. Prolonged slow growth in utero therefore seems to be followed by slow growth and development after birth.


Misdiagnosis and delayed diagnosis in traumatic intracranial haematoma. Out of 51 patients with traumatic intracranial haematoma admitted to a teaching hospital 11 (22%) died undiagnosed, and out of 307 such patients transferred to the West of Scotland Regional Neurosurgical Centre 111 (36%) had been deteriorating for more than 12 hours in another hospital. In two-thirds of these cases the delay was due to an erroneous diagnosis, either of cerebrovascular accident or of alcoholic intoxication.


Olfactory stimulation and olfactory bulb removal: effects on territorial aggression in male Syrian golden hamsters. The Syrian golden hamster exhibits territorial aggression in a laboratory setting, attacking much more when in the home cage than in the cage of another hamster. In expt. 1 it was found that female hamster vaginal discharge (FHVD) greatly inhibits the territorial aggression of males. In expt. 2 it was found that hamster territorial aggression is almost completely eliminated by bilateral olfactory bulbectomy but is unaffected by unilateral olfactory bulbectomy. However, bilateral olfactory bulbectomized (BOB) hamsters fought back normally if attacked first. Blinding increased the territorial aggression of male hamsters. In expt. 3 a peripheral means of impairing olfaction was used. The results suggest that it is the olfactory impairment, rather than the loss of some hypothetical non-sensory functions of the olfactory bulbs, that causes the elimination of territorial aggression after olfactory bulbectomy. Because of the extensive secondary and tertiary olfactory projections to the limbic forebrain and hypothalamus, and because most laboratory mammals are macro-osmatic, the nature and degree of olfactory control should be an important consideration in studies on the neural basis of social behavior.


Effects of morphine and naloxone on Renshaw cells and spinal interneurones in morphine dependent and non-dependent rats. The effects of microelectrophoretically administered morphine, naloxone, levorphanol and dextrorphan have been investigated on Renshaw cells and interneurones in the spinal cord of morphine-dependent and non-dependent anaesthetized rats. Morphine excited cholinoceptive neurones and enhanced the excitatory actins of acetylcholine and L-glutamate. This action of morphine appeared to be stereospecific and was antagonized by naloxone. Naloxone also antagonized acetylcholine-induced excitation but not L-glutamate-induced excitation. In dependent rats morphine was a more effective excitant of cholinoceptive neurones and naloxone was more effective as an antagonist of acetylcholine-induced excitations. These observations were interpreted as indicating that cholinergic mechanisms may be involved in morphine dependence and  naloxone-precipitated abstinence.


Histone-histone interactions. I. An electrophoretic study. Whole histone extracted from chromatin by either acid or protamine displacement was found by gel electrophoresis at pH7 to contain only two histone complexes, H2A-H2B and H3-H4, and uncomplexed histone H1. Although both complexes are dissociated at low pH or with high urea concentrations, removal of the denaturant resulted in complete complex reformation within minutes at the most. A syntematic investigation of binary, ternary and quaternary histone mixtures revealed that interactions also occur between histones H2B-H4 and H2A-H4. No evidence however was found for the formation of ternary and quaternary histone complexes.


Effects of alterations in maternal thyroid metabolism on embryonic thyroid development in the chick. The cause of embryonic thyroidal enlargement in the progeny of chickens fed goitrogenic rapeseed meal (RSM) has been investigated. It has been found that eggs from birds fed RSM are low in iodine. The low concentration of iodine in the eggs of the goitrous birds is apparently due to diversion of a large proportion of the circulating iodine into the thyroid glands with the result that the amounts reaching the developing ova are reduced.


Masson's "vegetant intravascular hemangioendothelioma:" a lesion often mistaken for angiosarcoma: study of seventeen cases located in the skin and soft tissues. The occurrence of Masson's "hemangio-endotheliome vegetant intravasculaire" (Masson's pseudoangiosarcoma) in the skin and soft tissues is illustrated with 17 surgically excised specimens. Two forms are recognized; it may appear either as a pure lesion or as a focal condition in a pre-existing vascular process, such as pyogenic granuloma or hemangioma. The clinical appearance is not specific and the diagnosis can only be established by microscopic examination. It shows a predilection for the head and extremities. Its characteristic morphologic appearance makes possible its differentiation from a group of benign and malignant vascular proliferations. The key microscopic feature is the presence of a papillary growth composed of hyperplastic endothelial cells supported by delicate fibrous stalks entirely confined within the vascular lumen. The lesion should not be mistaken for angiosarcoma, since its clinical behavior is invariably benign.


DNA synthesis of human, mouse, and rat mammary carcinomas in vitro: influence of insulin and prolactin. Carcinomatous mammary tissues, derived from six spontaneously arising mouse mammary tumors, six DMBA-induced rat mammary tumors, and 26 biopsy specimens of human breast tumors, were processed into slices and each tumor was inidvidually cultured for two days in Medium 199. The influence of bovine insulin (5.0 mug/ml) and ovine prolactin (10.0 mug/ml) on H3-thymidine incorporation into DNA was determined on the cultured tumor slices. Insulin consistenly (p less than 0.05-0.01) increased the incorporation of H3-thymidine into DNA of the organ cultures of mouse, rat, and human mammary carcinoma slices. The stimulatory effect of insulin was quantitatively more prominent in the mouse tumor slices than in the rat or human slices. The addition of prolaction to the insulin-containing culture medium further increased significantly (p less than 0.001) the incorporation of H3-thymidine into DNA of rat mammary carcinoma slices but had no significant effect on cultures of either mouse or human mammary carcinomas. The addition of prolactin to insulin and hydrocortisone-enriched medium containing slices of 20 individually cultured human breast carcinomas did not significantly influence the mean incorporation of H3-thymidine into DNA. However, a very small fraction (approximately equal to 15%) of these human breast carcinomas responded to prolactin by increasing the incorporation of H3-thymidine into DNA to a degree quantitatively comparable to the prolactin-sensitive, DMBA-induced rat mammary carcinoma. These results suggest that a very small fraction of human breast malignancies may respond to the growth-stimulatory effects of prolactin, but that the vast majority mimic more closely the prolactin-independent mouse mammary carcinoma.


Enzymes in peripheral and bone marrow serum in patients with cancer. Lactic dehydrogenase (LDH), glutamic-oxalacetic transaminase (GOT), and acid and alkaline phosphatase activities in bone marrow and in cubital vein serum were compared. For patients without cancer, marrow serum LDH attained levels four times as high, and GOT and alkaline phosphatase, levels twice as high as those normal for cubital vein serum; levels of acid phosphatase were the same for both sources. For patients with cancer, significant increase of enzyme levels over reference levels depends on the tumor origin and on the presence and localization of metastases. Marrow enzyme levels may become elevated with or without concurrent elevation in cubital vein serum. Concurrent elevations were found with colonic carcinoma and lymphoid leukemia, and noncurrent elevations, with prostatic cancer, myeloid leukemia, and myeloma. A nonconcurrent elevation of marrow enzymes indicates that the origin of the enzyme is in the marrow, whereas with concurrent elevation, the source of the enzyme may be another organ.


Hemangiosarcoma of liver and spleen treated by hepatic artery ligation, intraportal infusion chemotherapy, and splenectomy. Hepatic artery ligation is useful as a palliation of irressectable hepatic tumors, but does not always produce a satisfactory result. Hepatic tumor with high vascularity is expected to respond more favorably. In this context, primary liver cell carcinoma and carcinoid tumor or leiomyosarcoma of the liver have been satistfactorily treated by hepatic artery ligation. A case is presented of hemangiosarcoma of theliver and spleen treated effectively by hepatic artery ligation, splenectomy, and postoperative intraportal infusion of 5-fluorouracil, as indicated by the regression of hepatic tumors on postoperative scanning and arteriograpms.


Effect of l-asparaginase and asparagine deprivation on RNA metabolism in mouse leukemia L 5178Y cells in suspension culture. We analyzed the effect of asparagine starvation and L-asparaginase on RNA metabolism of mouse leukemia cell lines L5178Y, whose growth is dependent on the presence of asparagine, and L5178Y-R, whose growth is independent of the presence of asparagine. The deprivation of asparagine from the medium inhibited cellular protein synthesis by 30 to 40% of the control value in L5178Y cells, but not in L5178Y-R cells, whereas L-asparaginase inhibited synthesis by more than 80% in both L5178Y and L5178Y-R cells. The decrease in protein synthesis caused by asparagine starvation in L5178Y cells was accompanied by a decrease in ribosomal RNA synthesis. The synthesis of rRNA was also markedly blocked when L5178Y and L5178Y-R cells were exposed to L-asparaginase. The rate of synthesis of pulse-labeled RNA decreased significantly in the cells treated with L-asparaginase, and smaller pieces of polyadenylate containing pulse-labeled RNA (presumptive messenger RNA) appeared among monosomes and polysomes. However, the rate of messenger RNA synthesis was constant during asparagine starvation, and a marked accumulation of monosome was observed.


Major proteins and secretory character of the rough endoplasmic reticulum of a mouse mammary adenocarcinoma. The major protein composition and secretory character of rough microsomes isolated from the C3HBA mammary adenocarcinoma have been determined for the tumor borne in virgin female and lactating mice. Rough microsomes isolated from tumors have an equilibrium density distribution in a linear 1.0 to 2.0 M sucrose gradient similar to that exhibited by rough microsomes isolated from normal epithelium during lactation. Fractionation of the rough microsomal proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis reveals that, although the molecular weights of the major proteins are the same in normal and neoplastic rough microsomes, the relative levels at which these major proteins occur in neoplastic rough microsomes are distinct from those observed in normal rough microsomes isolated at various stages of differentiation. The most significant change in protein composition following neoplastic transformation is the presence of a high level of protein with a molecular weight of 200,000. There are changes in the relative levels of the major proteins of the rough endoplasmic reticulum attending transplantation of the tumor from virgin female to lactating mice; these changes, however, are quite different from those that occur in the rough endoplasmic reticulum of normal epithelium at the time of parturition. Rough microsomes isolated from tumors borne in virgin female mice discharge more than 90% of their nascent polypeptide chains extravesicularly upon premature termination by puromycin; there is no change in the vectorial discharge of these puromycyl peptides following implantation of the tumor into lactating mice. Thus, by a functional criterion, the rough endoplasmic reticulum of mammary adenocarcinoma cells appears similar to that found in normal, nonsecretory epithelium.


Effects of actinomycin D on cell cycle kinetics and the DNA of Chinese hamster and mouse mammary tumor cells cultivated in vitro. Actinomycin D produces changes in the cell cycle kinetics of Chinese hamster (HA1) and mouse mammary tumor cells (EMT6) cultivated in vitro. There was a reduced rate of progression of the cells through S phase and a G2 arrest, the duration and degree of which were drug dose dependent. The lethal effects of the drug on the two cell lines were comparable. At the molecular level, DNA single-strand breaks (true breaks and/or alkali-labile bonds) appear initially. After drug removal, the extent of disappearance of these DNA strand breaks was drug dose dependent. Similar results were obtained for the two cell lines. However, at the lower drug concentration (0.1 mug/ml), although the DNA strand breaks appeared to disappear after drug free incubation, about 60% of this cell population was not viable.


Hypoxia-dependent reduction of 1-(2-nitro-1-imidazolyl)-3-methoxy-2-propanol by Chinese hamster ovary cells and KHT tumor cells in vitro and in vivo. Incubation of Chinese hamster ovary cells and KHT murine fibrosarcoma tumor cells in the absence of oxygen with 1-[2-14C]nitro-1-imidazolyl)-3-methoxy-2-propanol, one of the most effective radiation sensitizers of hypoxic cells, results in the preferential reduction of 1-[2-14C]nitro-1-imidazolyl)-3-methoxy-2-propanol. The radioactivity associated with the acid-insoluble precipitate from cells incubated in nitrogen is about four times higher than that of cells incubated in air. When aqueous extracts of tissues of a C3H mouse bearing the KHT tumor, after i.p. injection with 1-[2-14C]nitro-1-imidazolyl)-3-methoxy-2-propanol, are analyzed, a reduction product is found in relatively higher yields in the tumor than in normal tissues. The relative radioactivity in the pellet from the tumor homogenate is also high in comparison with those of most normal tissues. These results provide suggestive evidence for a higher degree of hypoxic in the tumor than in most normal tissues. The formation of reduction products and their subsequent binding to macromolecules may explain the preferential toxicity of nitro compounds to mammalian cells under hypoxia conditions. These results suggest that some nitro compounds may be useful for the treatment of tumors having a high fraction of hypoxic cells even in the absence of radiation.


The effect of 1-beta-D-arabinofuranosylcytosine on cell viability, DNA synthesis, and chromatid breakage in synchronized hamster fibrosarcoma cells. Hamster fibrosarcoma cells were synchronized by mitotic selection and exposed to varying concentrations of 1-beta-D-arabinofuranosylcytosine (ara-C) for 2 hr in mid-S phase. There was a direct relationship between DNA synthesis inhibition and cytotoxicity produced by ara-C once DNA synthesis was decreased by over 85%. The noncytotoxic concentration of 10(-5) M ara-C produced little chromatid breakage; but extensive chromatid breakage and chromosomal rearrangement were seen in cells treated with the cytotoxic concentration of 10(-3) M ara-C, thus supporting earlier observations that chromatid breakage is highly correlated with cytotoxicity. Predominantly small DNA was synthesized when cells were treated with both 10(-5) and 10(-3) M ara-C, and this DNA could be completely chased into high-molecular-weight DNA after addition of deoxycytidine. Both concentrations of ara-C also inhibited, to different degrees, the joining of intermediate-size DNA fragments into larger DNA; thus neither parameter appeared directly related to the ara-C-produced cytotoxicity.


The effects of mercury on the structure and function of the hypothalamo-pituitary axis in the hamster. The neurons of the arcuate nucleus were studied ultrastructurally in female hamsters that were given saline or 1 mg mercuric chloride/day during one estrous cycle. Membranous whorls, composed of 4-26 concentric layers of smooth endoplasmic reticulum, were found in 7-8% of the neurons of control animals in diestrus (D3) and in proestrus (D3) and in proestrus (D4). Whorls were found in 5-16% of the neurons in mercury-treated animals on all days of the experimental period. The whorls appeared to be associated with RER and Golgi bodies. The levels of FSH and LH were measured in the plasma and pituitaries of both groups of animals. There was a significantly higher concentration of FSH in the pituitaries of experimental animals on D3 and of LH on D1C2 of the subsequent cycle; plasma levels of FSH and LH were similar in both groups throughout the experimental period. The relationship between the levels of gonadotropins and the presence of membranous whorls is discussed.


Mouse globin gene expression  in erythroid and non-erythroid tissues. Using cDNA hybridization, mouse globin RNA sequences can be detected in the nuclear and cytoplasmic RNA not only from erythroid cells (fetal liver and reticulocytes), but also in low amounts in non-erythroid tissues (adult brain and liver, and cultured lymphoma, untransformed, and transformed fibroblast cell lines). For the nuclear RNAs, melting curve, density, and size determinations on the hybrids confirmed the presence of an RNA species indistinguishable by cDNA hybridization from reticulocyte globin mRNA. Thus the genes coding for alpha- and beta-globin, proteins thought to be found only in a restricted range of differentiated cell types, may nonetheless be transcribed at very low levels in all mouse cells. The proportion of the globin RNA sequences in the nuclear RNA that are found associated with poly(A) sequences does not vary markedly from one tissue or cell line to another. In erythroid cells, however, the proportion of the cellular globin RNA sequences that are found in the cytoplasm is very much higher than in non-erythroid cells. This suggests the existence of messenger-specific nuclear  post-transcriptional control mechanisms which do not use poly(A) as a signal, and which allow the cytoplasmic accumulation of globin mRNA only in erythroid tissues.


Repeating units of Xenopus laevis oocyte-type 5S DNA are heterogeneous in length. The restriction enzymes Hind III and Hae III cleave Xenopus laevis 5S DNA at one and three sites, respectively, in each repeating unit of approximately 700 base pairs. The cleavage sites for both enzymes have been located within the repeating unit by denaturation mapping of the restriction fragments. The Hind III products and one of the Hae III fragments are variable in length, indicating heterogeneity in the length of the repeating unit in 5S DNA. This length heterogeneity is confined to the major A + T-rich spacer region. Repeating units differ from each other by discrete quanta of approximately 15 base pairs. The A + T-rich spacer has been shown to consist largely of tandem subrepeats of just this size (Brownlee, Cartwright, and Brown, 1974). We suggest that the repeat-length heterogeneity is due to variable numbers of these subrepeats in the spacer regions of the major repeating units.


Structural gene sets active in embryos and adult tissues of the sea urchin. Structural gene sequences active in a variety of sea urchin adult and embryo tissues are compared. A single-copy 3H-DNA fraction, termed mDNA, was isolated, which contains sequences complementary to the messenger RNA present on gastrula stage polysomes. Gastrula message sequences are 50 fold concentrated in the mDNA compared to total single-copy DNA. mDNA reactions were carried out with excess mRNA from blastula, pluteus, exogastrula, adult ovary, tubefoot, intestine, and coelomocytes, and with excess total mature oocyte RNA. A single-copy 3H-DNA fraction totally devoid of gastrula message sequences, termed null mDNA, was also reacted with these RNAs. Large differences in the extent of both mDNA and null mDNA reaction with the various RNAs were observed, indicating that in each state of differention a distinct set of structural genes is active, generally characterized by several thousand specific sequences. The complexity of gastrula mRNA was shown in previous work to be about 17 X 10(6) nucleotides. In units of 10(6) nucleotides, the complexities of the RNA sequence reacting with mDNA and with null mDNA in each tissue are, respectively, as follows: intestine mRNA: 2.1 and 3.7; coelomocyte mRNA: 3.5 and less than or equal to 1.4; tubefoot mRNA:  2.7 and less than or equal to 0.4; ovary mRNA: 13 and 6.7; oocyte total RNA: 17 and 20; blastula mRNA: 12 and 15; pluteus mRNA: 14 and less than or equal to 0.6; exogastrula mRNA: 14 and less than or equal to 0.6. The total complexity of each mRNA polulation is the sum of these values, as verified for several cases by reactions with total single-copy DNA. A relatively small set of mRNAs, the complexity of which is about 2.1 X 10(6) nucleotides, appears to be shared by several of the tissues studied.


A study of foldback DNA.

A study of foldback DNA. Nuclear DNA from eucaryotes contains a significant fraction which forms duplexes very rapidly and also independently of the DNA concentration. This fraction can be isolated by adsorption to hydroxylapatite and has been called foldback DNA (Britten and Smith, 1970). Here we extend previous studies to show that the foldback fraction is due to the existence of a finite number of foldback foci in each genome equivalent of DNA, approximately 10(5) in the case of Xenopus laevis. More significantly, we have isolated the foldback fraction in quantity from DNA of such a size (in one case broken randomly and in another digested with a restriction endonuclease) that only about 10% of the total DNA has foldback properties. If the foldback foci were located in precisely the same positions in all sets of the Xenopus laevis genome, the prediction would be that these foldback fractions would contain sequences representing 20% (random shear) and 10% (restriction endonuclease) of the total genome. In contrast, our results show that in both cases the foldback fraction contains the entire Xenopus laevis DNA sequence. One possible explanation of these observations is that as in procaryotes, eucaryotic DNA is randomly cross-linked. We show that cross-linkage of Xenopus laevis DNA is not sufficient to explain our observations. In consequence, we have adopted the hypothesis that the formation of foldback DNA is mainly an intrastrand phenomenon, but nevertheless occurs at different sites in different sets of the Xenopus laevis genome.


Cesium chloride gradients of chromatin after treatment with micrococcal nuclease. Cesium chloride equilibrium density centrifugation shows that treatment of rat liver nuclei with low concentrations of micrococcal nuclease for extremely short periods of time results in the appearance of chromatin fractions of low protein/DNA ratio and even free DNA. The DNA of these chromatin fractions is shorter than the DNA moiety of one chromatin subunit. The amount of high buoyant density material is decreased with increasing digestion time. We conclude that this material belongs to the minor chromatin fraction which is not organized according to the subunit model.


The organization of histones and DNA in chromatin: evidence for an arginine-rich histone kernel. We have examined the role played by various histones in the organization of the DNA of the nucleosome, using staphylococcal nuclease as a probe of DNA conformation. When this enzyme attacks chromatin, a series of fragments evenly spaced at 10 base pair intervals is generated, reflecting the  histone-DNA interactions within the nucleosome structure. To determine what contribution the various histones make to DNA organization, we have studied the staphylococcal nuclease digestion patterns of complexes of DNA with purified histones. Virtually all possible combinations of homogeneous histones were reconstituted onto DNA. Exhaustive digestion of a complex containing the four histones H2A, H2B,H3, and H4 yields a DNA fragment pattern very similar to that of whole chromatin. The only other combinations of histones capable of inducing chromatin-like DNA organization are H2A/H2B/H4 and those mixtures containing both H3 and H4. From an examination of the kinetics of digestion of H3/H4 reconstitutes, we conclude that although the other histones have a role in DNA organization within the nucleosome, the arginine-rich histone pair, H3/H4, can organize DNA segments the length of the nucleosome core in the absence of all other histones.


Glycoproteins of the plasma membrane of Dictyostelium discoideum during development. Differential binding of 125I-Con A to whole cells throughout the life cycle of dictyostelim discoideum indicates that the appearance of Con A binding proteins on the plasma membrane is under developmental regulation. Con A-Sepharose affinity chromatography of radio-iodinated plasma membrane preparations, followed by analysis with SDS-PAGE, revealed that there are at least 15 Con a binding proteins associated with the surface of 0 hr cells. Between 6 and 18 hr of development, the relative intensity of one of the bands, which corresponds to a protein of molecular weight of 150,000 daltons, increases dramatically.


The 5' leads to 3' polarity of the Xenopus Ribosomal RNA precursor molecule. Evidence is presented that in Xenopus laevis the 18S rRNA sequence is proximal to the 5' end of the rRNA precursor molecule and the 28S secquence is proximal to the 3' end. This assignment was made by digesting amplified ribosomal gene transcription complexes with EcoRI restriction endonuclease and spreading the cleaved transcription complexes for electron microscopy. From the known location of the EcoRI cutting sites within the transcribed region and the length of nascent chains on the cleaved transcription complexes, it was possible to make an unambiguous assignment of polarity.


Sequences of 5S ribosomal RNA from Xenopus mulleri and the evolution of 5S gene-coding sequences. Sequences for 5S RNA from somatic cells and oocytes of Xenopus mulleri are presented. Comparison with sequences previously given for Xenopus laevis indicates that the somatic 5S RNA genes of each species are more closely related to each other than either is to its own set of oocyte genes, suggesting that somatic and oocyte genes within each species are evolving independently. However, detailed analysis of sequence variants in each species suggests that there is a mechanism which allows occasional genetic exchanges between somatic and oocyte-specific genes. Possible genetic mechanisms which allows such an exchange are discussed.


[Malignant apudoma (argyrophilie carcinoma) of the uterine cervix in a 24-year-old woman in pregnancy]. Report on a case of the malignant APUDOMAP (argyrophil cell carcinoma) of the uterine cervix in a 24-years old women. The tumor appeared during the third pregnancy and fatal generalization resulted cca three month after operative delivery and panhysterectomy. Histologically argyrophil cells were proved both in the primary localization and distant metastases (liver, lymph nodes, ovaries, brain, meninges, skeleton). In the electron microscope of the bioptic specimen specific granules and microfilaments were identified in some tumour cells supporting the diagnosis of a cancer of the APUD-series. Some general features of primary cervical APUDOMAS also outlined.


Protein-bound carbohydrates in breast cancer. Liquid-chromatographic analysis for mannose, galactose, fucose, and sialic acid in serum. We describr high-resolution chromatographic analysis for protein-bound sialic acid in serum, with use of a cerate oxidimetric detector. Values for sera from normal women averaged 680.5 mg/liter, with a coefficient of variation of 23%. Including data obtained by previously developed chromatographic procedures for protein-bound mannose, galactose, and fucsoe, we assessed sera from breast-cancer patients whose malignancy had been categorized as either stable, responsive, or progressive (based on clinical observations spaced from two to five months apart). All of 12 responsive patients had decreases of protein-bound fucose averaging 34.5% (SD, 16.1) and all of 10 patients with progressive disease had increases averaging 38.3% (SD 21.5). Changes in fucose averaged less than 6.7% (SD, 4.9) for eight patients with clinically stable breast cancer. Changes in protein-bound mannose, galactose, and sialic acid did not correlate as well as did fucose with the clinical disease status of the patients.


Twins in the sibships and parental sibships of women with Turner's syndrome. The twin birth frequency in the sibships of 45 females with Turner's syndrome was 3.7% for the 21 females with 45,X and 5.7% for the 24 with other karyotypes. The 5.7% is significantly higher than the expected mean frequency of 1.4% in the Danish population from 1900 to 1974. The twin birth frequency in the sibships of the mothers of the 24 females with karyotypes other than 45 X was 4.7% which is also significantly higher than expected. The twin birth frequency in the sibships of the 21 mothers of 45,X females did not deviate significantly from the expected frequency, and nor did that of the father's sibships. It is concluded that these findings indicate that there might be an association between the increased risk of X chromosome aberration in the progeny due to mitotic non-disjunction, isochromosome or ring chromosome formation on one hand, and twin formation on the other hand. Such X chromosome aberration and twin formation might thus have some common aetiological factors, and some of the possibilities are discussed.


Inhibition of antibody-dependent cell-mediated cytotoxicity (ADCC) as a means of detection of immune complexes in the sera of patients with thyroid disorders and bronchogenic carcinoma. Circulating immune complexes in the sera of patients with thyroid disorders or bronchogenic carcinoma were investigated using an assay system based on the inhibition of the ADCC activity of rat spleen cells. Increased inhibition, as compared with that of the sera of age and sex matched controls, was found in patients with thashimoto thyroiditis, primary hypothyroidism and bronchogenic carcinoma. The degree of inhibition in the first two groups was markedly increased compared with control sera of the same IgG content. However, the results in the lung cancer group were slightly but not significantly greater than  in controls with the same level of IgG. Increased IgG levels were found in patients with thashimoto thyroiditis and thyrotoxicosis, and also in patients with bronchogenic  carcinoma who had mediastinal gland involvement. The lower level of sensitivity of the assay system was approximately 600 ng added aggregated IgG, corresponding to a concentration of 6mu g/ml in the sample assayed. It is possible that circulating immune complexes may exist in lung cancer, but at a level below that of the present assay system.


Recurrent stress incontinence of urine: preoperative assessment and surgical management. Surgical correction of stress incontinence in the female remains a complex and distressing problem. Just as there is no single etiologic factor present in all patients with stress incontinence, so ther is no single uniformly corrective operative procedure, as attested by the continued reports of new operative techniques or malfunctions of old procedures. Every therapeutic effort for stress incontinence, whether by means of medication, exercise, plication, suspension, sling, or some combination of these, effectively increases urethral resistance by tightening the musculofasical planes and supporting the urethra. Regrettably, none of these methods can be 100% effective. The patient who does not have good urethral smooth muscle, qualitatively and quantitatively, will remain incontinent. Regardless of its length, caliber, angle, or axis, a fixed, fibrotic, noncontractile urethra will not provide urinary control. I prefer to use the Marshall-Marchetti-Krantz procedure in patients who had previous operatons and have persistent or recurrent incontinence.


Enhancement of alcohol withdrawal convulsions in mice by haloperidol. Based on the data presented here and the clinical observations cited it would appear that although haloperidol has been used with a certain degree of success for the treatment of acute alcohol abstinence the authors would like to caution the clinician against widespread use of heloperidol for treatment of alcohol withdrawal. In experimentally induced ethanol withdrawal, chlordiazepoxide appears to be a more effective and safer agent for ameliorating symptoms associated with excitation such as tremor, insomnia, anxiety, and hyperexcitability. A double blind comparative clinical investigation between chlordiazepoxide and haloperidol for treatment of alcohol withdrawal is warranted.


Subunit structure of chromosomes in mitotic nuclei of physarum polycephalum. We have investigated the subunit structure of mitotic chromosomes of the acellular slime mould Physarum polycephalum, using the nuclease susceptibility of isolated mitotic nuclei as a probe. A characteristic pattern of DNA digestion products is obtained, containing approximately integral multiples of a basic 140 base pair DNA segment that resembles very closely the pattern in G2 phase nuclei of Physarum and of calf lymphocyte nuclei. These results demonstrate that during the process of chromosome condensation there is no alteration at the primary level of chromatin structure that is responsible for the characteristic DNA digestion pattern.


Manic depressive illness: onset, diurnal temperature and season of birth. Fifty-six carefully diagnosed manic-depressive patients were evaluated seeking a relationship between onset of the illness, diurnal temperature variation, and month and season of birth. With respect to diurnal temperature variation, we were able to discover no significant change in the difference between the high and low temperature as the time of onset of each episode of the illness approached, but there was a trend for the high-low temperature difference to decrease as time of onset approached. Occurrence of episodes was seasonally distributed with significantly more hospitalizations in winter and spring and fewer in summer and fall. The cohort of manic-depressive patients was not different from the Indiana population generally with respect to season of birth.


Lithium carbonate versus E.C.T. in the treatment of the manic state of identical twins with bipolar affective disease. Scientific evidence accumulated to date of the efficacy of lithium in the treatment and prophylaxis of bipolar affective states rests primarily on quantitative vector analysis and some double blind studies. To our knowledge, no clinical studies similar to this one exist in literature wherein indentical twins were subjected to the variables of electroconvulsive therapy and phenothiazines versus lithium carbonate and phenothiazines. The only other variable in the study was the hospital settings involved. The twins' course of treatment over a period of 12 years prior to their presentation wherein they received phenothiazines in various hospital settings is presented in detail. The current study would seem to suggest that E.C.T. and phenothiazines are superior to phenothiazines alone in shortening hospital course and perhaps in improving post-hospital adjustment. Further, prior to the advent of lithium, it may be evident that psychiatrists were not attuned to the diagnosis of affective states. Finally, lithium carbonate and phenothiazines, while requiring a significantly longer period of hospitalization than E.C.T. and phenothiazines, appear to be associated with preventing re-hospitalization for manic states, but a slightly decreased level of occupational adjustment. The author discusses the potential value of these patients in determining the clinical efficacy of future drugs in bipolar affective states, as well as the necessity to obtain future studies of the creatinine phosphokinase level.


[Demonstration of hepatitis-B(surface) antigen with the Hepatest: a new passive haemagglutination test (author's transl)]. The Hepatest, a passive haemagglutination test, proved to be, like other tests of this group, significantly more sensitive for routine examination of hepatitis B(surface) antigen than tests of the first and second generation, but was not as sensitive as the Ausria II 125 test. The lowest HBs antigen concentration demonstrable in the Hepatest was 31 ng/ml for the subtype ad and 47 ng/ml for subtype ay (in the Ausria II 125 test lowest concentrations were 1.0 and 1.5 ng/ml, respectively). There were several false-positive reactions in the specificity test. Thus all positive reactions would need to be checked, more often than with the Ausria test. But the Hepatest proved to be the most practicable of all the third generation tests, both with regard to shortness of reaction time, technical requirements and ease of reading off the results.


Erythropoietin production by human renal carcinoma cells in culture. Cells from human renal tumors were grown in monolayer cultures, and the media obtained at each medium change were assayed for erythropoietin activity. The medium from carcinoma I (a granular cell tumor) contained a high level of activity initially. The concentration of erythropoietin activity decreased with time in culture, but was significantly higher than that in controls after four months in vitro. There was , in addition, evidence of an inhibitory material present in the culture media. The activity formed by the tumor cells could be neutralized by an antibody to human urinary erythropoietin. The difference between activity measured in marrow cell cultures and that found by in vitro assay, and the chromatographic properties of the active preparation, suggest that the tumor-derived activity may be largely asialoerthropoietin. Two other renal carcinomas, of a different cellular type, produced significant erythropoietic activity.


Studies on the role of the preoptic area in the control of reproductive function in the rat. The present study was designed to determine the effects of bilateral lesions restricted to the medial preoptic area on reproductive function in rats. Adult female rats were placed in a stereotaxic instrument, and bilateral lesions were made in the medial preoptic area of the diencephalon using an anodic current and platinum or stainless steel electrodes. Both types of lesions produced identical effects. Most animals with lesions restricted to the medial preoptic area showed repeated periods of pseudopregnancy for the duration of their lives. Ova were detected in the oviducts on the day of estrus between pseudopregnancies, and deciduomata could be produced by uterine traumatization during pseudopregnancy. Daily administration of lergotrile mesylate, a dopamine agonist, for about two weeks, to the pseudopregnant rats with lesions resulted in the appearance of normal 4 or 5-day estrous cycles. These results suggest that the medial preoptic area may not be necessary for ovulation in the rat, and that the normal function of the preoptic area in the control of reproductive cyclicity may be mediated via dopaminergic neurons. In agreement with earlier studies, we found that lesions that extended into the anterior hypothalamus resulted in constant estrus.


Ovarian regulation of postcoital gonadotropin release in the rabbit: reexamination of a functional role for 20 alphadihydroprogesterone. In the rabbit, it has proposed that an ovarian progestin, 20 alpha-dihydroprogesterone (20alphaP), released at mating, is essential for a normal postcoital LH surge. However, we measured plasma levels of LH and 20alphaP after mating in rabbits and observed that the frequency, magnitude and time-course of changes in circulating levels of 20alphaP seemed inappropiate to account for the rapid and major surge of LH secretion. This prompted us to re-evalute the role of the ovary in regulating postcoital LH secretion. In chronically ovariectomized (greater than 30 days) does pretreated with estrogen, mating induced a normal LH surge in only 1 of 10 animals, indicating that an ovarian product in addition to estrogen is required for a normal postcoital LH surge. However, when 20alphaP was injected soon after mating in chronically ovariectomized does pretreated with estrogen, only 2 of 9 displayed normal LH surges; this proportion is not different from that (1/10) observed with estrogen treatment alone. To demonstrate that the estrogen treatment, which produced supraphysiologic plasma estradiol levels, did not itself block LH release, 6 intact anestrus females were treated with the same estrogen regimen. Estrus was induced in 5 and each displayed a large post-coital LH surge and ovulated. As a final test of the 20alphaP hypothesis, 5 spontaneously estrous does were ovariectomized within 15 min post coitum to abolish acute increases in circulating ovarian hormones. Three animals released LH in amounts and temporal pattern indistinguishable from intact estrous does. A fourth released smaller amounts of LH. Two of 4 sham-operated does also had normal LH surges. These findings indicate that ovarian hormones are required before mating to support the capacity of the LH secretory mechanism to respond to coitus. Chronic alteration in the hormonal milieu by ovariectomy appears to produce a change in the hypothalamo-hypophysial complex that is not reversed by estrogen, alone. More importantly, these results demonstrate clearly that neither 20alphaP nor any other ovarian hormone is required post coitum, at least after 15 min, for normal LH release.


Changes in body weight, milk production, food and water consumptions and vaginal smears in rats during prolonged lactation. Concurrent observations were made in the rat on the changes in the milk production, body and organ weight, food and water consumptions and in the ovarian function in two separate series of experiments in which the period of lactation was prolonged to 60 and 45 days, respectively, by replacing suckling pups by younger ones. In most of the variable examined, marked changes occured between day 20 and 30 of lactation, that is, at the stage correspondong to the end of the normal lactation period. Milk production rate during prolonged lactation expressed by daily gain in weight of litters decreased to 60% of the level before day 20, and was associated with a great reduction of nucleic acid contents of the mammary gland. Definite decreases in weight of the anterior pituitary and the adrenal glands were observed. The food intake remained constant from day 15 of lactation onwards, therefore, the feed efficiency for milk production declined gradually during the period of normal lactatation remaining at a low level thereafter.. After the first recurrence of vaginal estrus which also occured between day 20 and 30, the replacement of litters was followed by the appearance of estrus, whereas replacement before day 15 did not affect the ovarian function. In addtition to these changes, a depressing effect of estrus on the milk production was observed.


Duration of diestrous period and secretion of progestins during prolonged lactation in the rat. The duration of the diestrous period in vaginal smears and the levels of progestins in ovarian venous blood were investigated in rats whose lactation period was prolonged by replacing pups at appropriate intervals. In prolonged lactating rats, the first diestrous period after the pospartum ovulation (21.7 +/- 0.8 days) was similar to that occurring in normal lactating rats. Subsequently proestrous or estrous vaginal smears reappeared at intervals during prolonged lactation. A total of 19 rats were studied during this period of prolonged lactation and 44 diestrous periods were observed. Forty-eight % of these periods ranged in length from 11 to 16 days (mean of 12.7). Very short diestrous periods lasting less than 3 days were observed 25% of the time. Diestrous periods lasting more than 17 days were observed only infrequently (7%). Progesterone concentration in the ovarian vein during the 2nd diestrous period reached a peak 4 days after the 1st estrus and the level was maintained until 11 days after the 1st estrus. The level of 20alpha-hydroxypregn-4-en-3-one remained fairly constant throughout the 2nd diestrous period.


Recurrence of estrous cycles in the light-induced persistent estrous rats by reserpine. Effect of continuous illumination (LL) on ovarian functions was compared at various ages between the rats exposed to LL from 70 days of age (LL-70) and exposed to LL from the day of birth (LL-0). LL-0 rats retained corpora lutea in their ovaries in higher incidence than LL-70 rats. Effect of reserpine on the restoration of cyclicity was studied in LL-0 and LL-70 rats shortly after they became persistent-estrous. LL-0 rats restored cyclicity in higher incidence than in LL-70 rats.


In vitro transfer rate of 14C from acetate-1-14C into ovarian steroids in the rat ovary during the estrous cycle and effects of LH and FSH. Fluctuations of ovarian biosynthetic activity and effects of exogenous LH and FSH on it during the estrous cycle were investigated by measuring in vitro transfer rates of 14C from 14C-1-acetate into progesterone (P), 20 alpha-hydroxy-pregn-4-en-3-one (20 alpha-OH-P) and estrogen (estradiol and estrone, E) in the ovarian homogenates from rats autopsied at 2 hour intervals. The transfer rate of 14C from 14C-1-acetate into P was lowest in the afternoon of estrus and increased from the morning of diestrus 1, making its peaks during the afternoon of diestrus 2 and in the midnight of proestrus. The transfer of 14C into 20 alpha-OH-P was high on the days of diestrus 2 and proestrus with its peak in the afternoon of the latter day. The maximum transfer of 14C into E in the afternoon of proestrus and a high rate in the morning of estrus with relatively low one in the midnight were observed. Exogenously injected LH (150 mug) or FSH (150 mug) was either stimulatory or inhiibitory to the transfer rates of 14C from 14C-1-acetate into ovarian steroids. During day time of diestrus 2 and midnight between proestrus and estrus, the transfer of 14C into P and 20 alpha-OH-P increased by LH, and during day time of proestrus and from the afternoon of estrus to the morning of diestrus 1 decreased. The transfer of 14C into E increased by LH from the afternoon of diestrus 2 to the morning of proestrus, and decreased during the afternoon of proestrus and from the afternoon of estrus to the morning of diestrus 2. Administration of FSH was also stimulatory or inhibitory. The 14C transfer into P and 20 alpha-OH-P increased by FSH from the afternoon of estrus to the morning of proestrus, but in the afternoon of proestrus they decreased. Transfer of 14C into E increased by FSH significantly on the days of diestrus 2 and proestrus, and slightly on the day of estrus, while it decreased in the afternoon of diestrus 1.


Rostral displacement of the palatopharyngeal arch: a case report. A horse with rostral displacement of the palatopharyngeal arch was found to have a bilaterally symmetrical deformity of the laryngeal area. Both left and right cricopharyngeal muscles were absent. The shape of the thyroid cartilage was grossly abnormal and vestiges of the cricothyroid muscles were attached only to the cricoid cartilage. It was suggested that such an anomaly could have resulted from aberrant development of the fourth branchial arch.


Urethral sphincterotomy for delivery of vesical calculus in the mare: a case report. A case of vesical calculus in which haematuria was not a presenting sign is described. Methods of removal of vesical calculi are discussed. In this case, urethral sphincterotomy facilitated delivery.


Dopamine receptors: selective agonists and antagonists of functionally distinct types within the feline brain. Unilateral injections of agents that are useful tools for differentiating electrophysiologically distinct dopamine receptors within the snail Helix aspersa are given into behaviourally distinct dopamine sensitive areas within the caudate nucleus of cats. Dopamine-elicited contralateral head-turning is selectively mimicked by apomorphine and selectively inhibited by haloperidol, whereas dopamine-elicited homolateral head-turning including oro-facial dyskinesias is selectively mimicked by (3, 4-dihydroxy-phenylamino)-2-imidazoline (DPI) and selectively inhibited by ergometrine, piribedil and noradrenaline. These results indicate that the caudate nucleus of cats contains two functionally and pharmalogically distinct dopamine receptors and that Helix aspersa dopamine sensitive neurons can be used as model for the design of drugs selectively interfering with each type of receptor. The implications of these findings are considered in view of the efficacy of piribedil, L-DOPA and neuroleptics in psychomotor diseases in man.


Ergot alkaloids as dopamine agonists: comparison in two rodent models. A series of ergot alkaloids, together with the DA agonists apomorphine and piribedil, were tested for protective effects against audiogenic seizures in an inbred strain of mice (DBA/2) and for induction of circling behaviour in mice with unilateral destruction of one nigrostriatal DA pathway. The order of potency against audiogenic seizures was apomorphine greater than ergocornine greater than bromocryptine greater than ergometrine greater than LSD greater than methysergide greater than piribedil while that observed in the rotating mouse model was apomorphine greater than ergometrine greater than ergocornine greater than bromocryptine greater than piribedil. LSD caused only weak circling behaviour even when administered in high doses (greater than 1 mg/kg). Methysergide was ineffective. Prior administration of the neuroleptic agent haloperidol blocked the effect of DA agonists and of ergot alkaloids in both animal models. The possible action of ergot alkaloids as DA agonists is discussed.


Circling behavior in rats with 6-hydroxydopamine or electrolytic nigral lesions,. Unilateral destruction of dopaminergic cell bodies in the substantia nigra zona compacta (SNC) was performed in rats using either electrocoagulation or chemical lesioning (6-hydroxydopamine, 6-OHDA). Neostriatal dopamine concentration ipsilateral to an electrolytic lesion was 34% of the contralateral side 2-3 weeks after operation; serotonin and noradrenaline brain levels were not altered. In contrast, dopamine and noradrenaline forebrain concentrations ipsilateral to a 6-OHDA lesion were 20 and 31%, respectively, of the contralateral side. After 6-OHDA, dopamine concentrations in the ipsilateral neostriatum were reduced to levels below the sensitivity of the fluorometric assay; cortical, brainstem and neostriatal serotonin levels, on the other hand, were not altered after 6-OHDA. Ipsilateral circling behavior was elicited by d-amphetamine after electrolytic and chemical lesioning. In contrast, the direction of circling produced after apomorphine differed between the two lesion models: contralateral circling behavior was exhibited by 6-OHDA-lesioned rats, whereas ipsilateral circling was produced in animals with electrolytic lesions. Contralateral circling was induced in both lesion-type models by haloperidol or pimozide. S.c. atropine administration induced ipsilateral circling in rats with 6-OHDA lesions, whereas contralateral circling was observed after arecoline. Animals with electrolytic SNC lesions turn ipsilaterally after s.c. administrations of either arecoline or atropine. The data indicate that the electrolytic and 6-OHDA circling behavior models represent two different neuropharmacological states and it is, therefore, suggested that comparisons of data obtained from models using different methods of lesioning be made with caution.


Dopaminergic-cholinergic interactions in naloxone-induced circling in morphine-dependent rats with nigral lesions. 3-4 weeks after placement of a unilateral, electrolytic lesion of the substantia nigra zona compacta, rats were highly dependent on morphine by the s.c. morphine pellet implantation technique. Following challenge with a supramaximal naloxone dose of 20 mg/kg i.p., both continuous contralateral circling behavior and severe withdrawal signs in morphine-dependent, lesioned rats were elicited. After various drug pretreatments, the contralaeral circling behavior precipitated by naloxone was: (a) reversed to ipsilateral circling by i.p. apomorphine or d-amphetamine, (b) unaltered by i.p. haloperidol or intraneostriatal arecoline administered into the intact neostriatum, and (c) reversed to ipsilateral circling by the administration of atropine into the intact neostriatum. Atropine, apomorphine and amphetamine all interfered with the manifestation of naloxone-precipitated abstinence. These data suggest that a diminution of dopaminergic or an enhancement of cholinergic activities, or both, occur at the level of the neostriatum during naloxone-precipitated withdrawal in morphine-dependent rats.


Effect of pargyline on morphine tolerance and physical dependence development in mice. The effects of single and repeated pargyline administration on morphine antinociception in both naive and morphine-tolerant mice and on naloxone-precipitated withdrawal in morphine tolerant-dependent animals were investigated. Adult, male Swiss-Webster mice were rendered tolerant to and dependent on morphine by the s.c. pellet implantion technique. Morphine analgesia, as assessed by the tail-flick antinociceptive test, was potentiated in tolerant animals by acute adminstration of pargyline but antagonized by repeated pargyline administration; pargyline produced similar effects in non-tolerant mice and to the same relative degree. Repeated pargyline treatment during morphine pellet implantation enhanced the withdrawal jumping response precipitated by naloxone in dependent mice. Pargyline also, after a single injection, exacerbated jumping in mice undergoing abrupt withdrawal. Neither acute nor chronic pargyline administration altered the brain distribution of injected morphine in non-tolerant mice. It was concluded that pargyline may modify acute morphine actions and withdrawal without materially altering the process(es) involved in the development of tolerance and physical dependence.


Methylphenidate-like effects of the new antidepressant drug nomifensine (HOE 984). Nomifensine (HOE 984) belongs to a chemically new class of drugs with reported antidepressant properties. Nomifensine, like methylphemidate, d-amphetamine and apomorphine, induces strong, intense stereotypes behaviour in the rat. The nomifensine-induced stereotyped behaviour was completely antagonized by pretreatment with reserpine (7.5 mg/kg, 18 h) but not by short-time pretreatment with alpha-methyltyrosine (250 mg/kg, 2 h.) Nomifensine thus differs from d-amphetamine and apomorphine but resembles methylphenidate on stereotyped behaviour. Nominfensine, M1 (8-amino-2-methyl-4-(4-hydroxyphenyl)-1,2,3,4-tetrahydroisoquinoline fumarate) (Hoechst), methylphenidate and d-amphetamine induced a strong increase in the brain level of homovanillec acid (HVA), whereas the dopamine uptake inhibitor benztropine induced no changes in HVA and cocaine induced only a small increase. Nomifensine and the M1 metabolite, like methylphenidate, also increased 3,4-dihydroxyphenylacetic acid (DOPAC) whereas amphetamine, apomorphine, benztropine and cocaine decreased this dopamine metabolite. This suggests that the stereotyped licking and/or biting activities in the rat are related to dopamine releasing properties of nomifensine, methylphenidate and amphetamine. This is further supported by an inverse relationship between the in vitro dopamine uptake inhibitory concentrations and the sterotypy-inducing dose levels of nomifensine and d-amphetamine. Amphetamine caused a strong, and nomifensine and apormorphine a week increase in brain 3-methoxy-4-hydroxyphenylglycol (MOPEG).


On the origin of the protein yolk in the oocytes of Aplysia depilans (Gastropoda, Opistobranchia). In the oocytes of Aplysia depilans it has been observed electronmicroscopically that "gemmae" or "daughter-nucleoli" separate from the nucleolus. These "gemmae" after having overcome the nuclear membrane, would aggregate together, even after transformation, into yolk globules. These observations seem to give plausibility to the hypothesis that there is a close relationship between production of ribonucleoproteic material by the nucleolus and yolk production.


[A progesterone-dependent pheromone of the female mouse (author's transl)]. Female mice kept in groups exhibit less oestrous smears than females kept singly. Ovariectomized females have only slightly reduced number of oestrous smears, but ovariectomized females injected with progesterone act on other females like intact ones. It is concluded that a progesterone-dependent pheromone excreted by dioestrous females acts oestrus-depressing on other females.