Developmental control of messenger RNA for hepatic tryptophan 2,3-dioxygenase. The enzyme tryptophan 2,3-dioxygenase [EC 1.13.11.11; L-tryptophan:oxygen 2,3-oxidoreductase (decyclizing)] first appears in the livers of young rats around the 15th postnatal day, and increases to the adult level by the 22nd day. Studies have shown that the appearance and subsequent development of the enzyme activity result from an increase in the rate of its synthesis and thus in the amount present in the liver. In this study, we have attempted to ascertain whether the appearance and development of tryptophan 2,3-dioxygenase mRNA coincided with, and thus led to, the development of enzyme activity, or whether the biosynthesis of this enzyme was due to a developmental event enabling translation of a preexisting, sequestered, reservoir of its mRNA. Using a cell-free protein-synthesizing system based on a wheat germ S30 supernatant, we measured the level of tryptophan 2,3-dioxygenase mRNA in the livers of rats between 0 and 22 days of age. We found that functional tryptophan 2,3-dioxygenase mRNA is not detectable in rat liver until the 15th postnatal day. It increases to the adult level by the 22nd postnatal day, in parallel with the enzyme. The appearance and development of tryptophan 2,3-dioxygenase are the direct consequence of the parallel appearance and development of its mRNA. It has been shown that glucocorticoids, which induce tryptophan 2,3-dioxygenase activity in adult rats, are capable of inducing the appearance of this enzyme precociously in 8- and 10-day-old rats. We have found that it is also possible to induce tryptophan 2,3-dioxygenase catalytic activity with hydrocortisone precociously in 4-day-old rats. Moreover, precocious induction of enzyme activity and the induction that occurs during the enzyme's normal developmental rise to the adult level between 15 and 22 days, are mediated through parallel increases in the level of tryptophan 2,3-dioxygenase mRNA. The present findings indicate that glucocorticoids are developmental hormones that act upon the postnatal hepatocyte to evoke elevated levels of the mRNA species coding for tryptophan 2,3-dioxygenase; the findings are compatible with the hypothesis that such hormones act by initiating and accelerating transcription of the structural genes coding for the alpha and beta protomers of this enzyme.


Sites of transcription initiation in vivo on Xenopus laevis ribosomal DNA. We report the results of a novel method for locating sites of transcription initiation using a complex of capping enzymes from vaccinia virions that catalyze the reaction pppG + S-adenosylmethionine + (p)ppXpYpZp..... -->(7m)GpppXpYpZp..... [Ensinger, M. J., Martin, S. A., Paoletti, E. and Moss, B. (1975) Proc. Natl. Acad. Sci. USA 72, 2525-2529]. This enzyme complex will cap di- or triphosphate termini but will not cap monophosphate or hydroxyl termini. Xenopus laevis 40S precursor rRNA from oocytes is capped by these enzymes, and we conclude that it has 5'-polyphosphate termini. Therefore, 40S RNA must represent the primary transcript of amplified X. laevis ribosomal DNA. The majority of 40S molecules with polyphosphate termini begin with the sequence (p)ppAAG. There is evidence, however, that the 5' terminus may be heterogeneous. The majority of all detectable initiation events were localized close to the region coding for the 5' end of the 40S RNA. No initiation sites were detected in the nontranscribed spacer, but an apparent initiation site in the middle of the transcribed region was also observed.


Distribution of rat liver albumin mRNA membrane-bound and free in polyribosomes as determined by molecular hybridization. Recently, we purified rat liver albumin mRNA and prepared albumin [(3)H]cDNA. Using albumin [(3)H]cDNA in molecular hybridization experiments, we have now determined the distribution of albumin mRNA sequences in membrane-bound and free liver polyribosomes prepared by techniques in which there is high recovery of polyribosomes without evidence of degradation. By using molecular hybridization to measure specific mRNA sequence content, numerous problems could be avoided in interpretation of results as obtained by cell-free protein synthesis or immunological methods. Under these conditions, 98% of albumin mRNA sequences in polyribosomes are found in the membrane-bound fraction.


Defects of glycosyltransferase activities in human fibroblasts of Pk and p blood group phenotypes. We demonstrate that human fibroblasts of the rare Pk phenotype lack globoside, which was identified as the blood group P antigen, and that p cells possess neither globoside nor trihexosyl ceramide, which was identified as Pk antigen. Our investigations indicate also that these glycosphingolipid patterns are most likely caused by inherited preferential biosynthetic pathways in the abnormal phenotypes rather than by excess catabolism of the antigens. Evidence is presented that the fibroblasts of Pk phenotype lack beta-N-acetylgalactosaminyltransferase (globoside synthetase; UDP-N-acetylgalactosamine:trihexosylceramide beta-N-acetylgalactosaminyltransferase; EC 2.4.1.79) activity, and those of p are deficient in alpha-galactosyltransferase (trihexosylceramide synthetase; UDP galactose:lactosylceramide alpha-galactosyltransferase) and possibly also in globoside synthetase. The diminished globoside synthetase activity in p cells, however, is not caused by the defect in the gene coding for this enzyme. It appears, rather, to be caused by a failure in gene expression because one-third of Pk X p hybrids became able to express P antigenicity with a time lag of 3-4 days after cell fusion [Fellous, M., Gerbal, A., Nobillot, G. & Weils, J. (1977) Vox Sang. 32, 262-268].


High resolution two-dimensional electrophoresis of human plasma proteins. The two-dimensional electrophoretic technique of O'Farrell has been adapted to the analysis of human plasma proteins, and 30 polypeptides have been identified in the pattern produced. Genetic variants involving charge (isoelectric point) or size (molecular weight in the presence of sodium dodecyl sulfate) changes should be routinely detectable in at least 20 proteins at once, facilitating studies of human mutation rates.


Orientation of bacteriorhodopsin in Halobacterium halobium as studied by selective proteolysis. The orientation of bacteriorhodopsin in the purple membrane of Halobacterium halobium has been studied by proteolytic degradation of purple membrane sheets, reconstituted vesicles, and whole cells, with the following results: (i) Bacteriorhodopsin in purple membrane sheets is cleaved at a single site by Pronase or trypsin; a polypeptide segment of about 15 amino acids is lost from the carboxyl end. Carboxypeptidase A sequentially releases amino acids from the carboxyl end; the tetrapeptide sequence -Ala-Ala-Thr-Ser(COOH) was tentatively deduced for this terminus. (ii) The apomembrane, which lacks retinal, undergoes a second cleavage with trypsin releasing a fragment of approximately 6300 molecular weight from the amino terminus. (iii) Vesicles reconstituted from the purple membrane sheets and synthetic lecithins, in which the direction of proton pumping is opposite to that in the whole cells, have the carboxyl terminus of bacteriorhodopsin accessible to proteolysis. (iv) In envelope vesicles, which largely pump protons in the same direction as the whole cells, the carboxyl terminus is largely protected against proteolysis. (v) Treatment of whole cells with proteinase K hydrolyzes the cell wall proteins but has no effect on acteriorhodopsin. However, the same treatment after lysis of the cells results in degradation of the hydrophilic region at the carboxyl terminus. The results show that the carboxyl terminus as well as the additional cleavage site near the amino terminus observed in apomembrane are on the cytoplasmic side of the purple membrane.


Oligopeptides as potential antiaggregation agents for deoxyhemoglobin S. Oligopeptides that mimic segments of the amino acid sequence of hemoglobin S at potential contact sites can be used to inhibit aggregation. These oligopeptide inhibitors raise the minimum gelling concentration of deoxyhemoglobin S so that chemical modification does not have to be used. The hexapeptide amides of both betaS 1-6 which is believed to be one of the contact areas among aggregates, and betaA 1-6 of hemoglobin A increase the minimum gelling concentration by more than 70%. The hexapeptide amide beta79-84 behaves like beta1-6 (beta being betaS or betaA). Shorter oligopeptides, such as betaS3-6, are less effective as an inhibitor but longer ones, such as betaS1-8, are no more effective than beta1-6. Permutations of the sequence, such as betaS125634, do not alter the percent increase in the minimum gelling concentration. Leu- and Met-enkephalin increase the minimum gelling concentration just as beta1-6 does, but (Pro)6 is not very effective. Thus, the use of complementary oligopeptides as inhibitors is extended to include certain "flexible" peptides, which can adapt themselves to interfere with the molecular contacts and thereby gelation of deoxyhemoglobin S.


The bicistronic nature of lens alpha-crystallin 14S mRNA. The A2 and B2 polypeptide chains of calf lens alpha-crystallin are synthesized on a 14S, 1500-nucleotide mRNA and a 10S, 735-nucleotide mRNA, respectively. The 10S mRNA is theoretically compatible with the size of the B2 chain, but the 14S mRNA contains approximately twice the required number of nucleotides necessary for A2 chain synthesis. This fact raises the question of the function of the additional nucleotide sequence in the 14S mRNA. The following observations on 14S mRNA suggest that it may contain an additional cistron. (i) Under a number of denaturing conditions, 14S mRNA continues to retain its initial size characteristics. (ii) In addition to synthesis of the A2 chain, 14S mRNA directs the synthesis of another polypeptide with the same electrophoretic mobility as that of the B2 chain. (iii) Molecular hybridization of the 14S mRNA with the cDNA produced from the 10S mRNA suggests that 2 mol of the cDNA bind to 1 mol of the 14S mRNA. (iv) Examination of the nucleotide sequences of the 10S and 14S mRNAs by two-dimensional maps of RNase A and T1 digests indicates marked similarity. The overall data suggest that the additional cistronic component may carry coding information for an alpha-crystallin polypeptide or a closely related polypeptide species.


DNA sequencing with chain-terminating inhibitors. A new method for determining nucleotide sequences in DNA is described. It is similar to the "plus and minus" method [Sanger, F. & Coulson, A. R. (1975) J. Mol. Biol. 94, 441-448] but makes use of the 2',3'-dideoxy and arabinonucleoside analogues of the normal deoxynucleoside triphosphates, which act as specific chain-terminating inhibitors of DNA polymerase. The technique has been applied to the DNA of bacteriophage varphiX174 and is more rapid and more accurate than either the plus or the minus method.


Hepatic mitochondrial cytochrome P-450: isolation and functional characterization. A CO-binding heme protein was solubilized and partially purified from the inner membrane fraction of rat liver mitochondria by a modification of a method [Imai, Y. & Sato, R. (1974) Biochem. Biophys. Res. Commun. 60, 8-14] developed to purify cytochrome P-450 from liver microsomes. The partially purified preparation contained protoheme and its spectral properties are characteristic of the heme proteins of the cytochrome P-450 family. The isolated cytochrome P-450 preparation could reconstitute a CO-sensitive, NADPH-dependent 26-hydroxylation activity for 5beta-cholestane-3alpha,7alpha,-12alpha-triol when supplemented with NADPH-adrenodoxin reductase and adrenodoxin, both purified from bovine adrenocortical mitochondria. Unlike a cytochrome P-450 purified from liver microsomes of drug-untreated rats, however, the liver mitochondrial cytochrome P-450 could not catalyze NADPH-dependent benzphetamine N-demethylation in the presence of adrenodoxin reductase and adrenodoxin or function with the purified microsomal NADPH-cytochrome c reductase plus Emulgen 913 as an electron-donating system. It is concluded that the rat liver inner mitochondrial membrane houses a species of cytochrome P-450 functional in 5beta-cholestane-3alpha,7alpha,12alpha-triol 26-hydroxylation.


Two distinct adenosine-sensitive sites on adenylate cyclase. The effects of adenosine and adenosine analogs on adenylate cyclases from several tissues have been examined. Two adenosine-reactive sites have been identified: (i) the "R" site, occupancy of which usually leads to activation of cyclase and which requires integrity of the ribose ring for activity, and (ii) the "P" site, which mediates inhibition and requires integrity of the purine ring for activity. Biphasic effects of adenosine are explained by the presence of both sites on a single adenylate cyclase. Comparison of these data with those in the literature indicates that adenosine-reactive "P" and "R" sites are present generally.


Presence of protein A24 in rat liver nucleosomes. Two-dimensional gel profiles of the 0.2 M H2SO4-soluble proteins of monomer nucleosomal fractions were found to contain protein A24. Protein A24 is of interest because it is composed of histone 2A and "ubiquitin", apparently joined by an isopeptide linkage [Goldknopf, I.L. & Busch, H. (1977) Proc. Natl. Acad. Sci. USA 74, 864-868; Hunt, L.T. & Dayhoff, M.O. (1977) Biochem. Biophys. Res. Commun. 74, 650-655]. Monomer nucleosomal fractions were obtained by sucrose density gradient centrifugation of micrococcal nuclease digests of rat liver nuclei. As shown by their DNA size, the monomer fractions were highly purified. Proteins A24 and Bu, another protein of unknown characteristics, were found along with histones 1, 2A, 2B, 3, and 4 in the monomer fractions in relative amounts similar to those found in extracts from whole nuclei and chromatin. Other acid-soluble proteins found in the nuclear and chromatin extracts were essentially absent from the monomer fraction. Inasmuch as protein A24 and Bu were found in lesser amounts than the histones, it is suggested that they are associated with specialized subsets of nucleosomes.


Alternative aspirins as antisickling agents: acetyl-3,5-dibromosalicylic acid. Acetyl-3,5-dibromosalicylic acid (dibromoaspirin) is shown to be a potent acylating agent of intracellular hemoglobin in vitro. Transfer of the actyl group of dibromoaspirin to amino groups of hemoglobins A and S seems to occur predominantly at just two or three sites on these proteins. This acetylation produces moderate increases in the oxygen affinities of normal and sickle erythrocytes. Furthermore, treatment of intracellular hemoglobin S with dibromoaspirin directly inhibits erythrocyte sickling. This antisickling effect is paralleled by an increase in the minimum gelling concentration of deoxy hemoglobin S extracted from sickle erythrocytes that had been exposed to low concentrations of dibromoaspirin. These observations suggest that dibromoaspirin might be an effective antisickling agent in vivo.


Nucleotide sequence of 5' terminus of alfalfa mosaic virus RNA 4 leading into coat protein cistron. The sequence of the 5'-terminal 74 nucleotides of alfalfa mosaic virus RNA 4, the mRNA for the viral coat protein, has been deduced by using various new techniques for labeling the RNA at the 5' end with 32P and for sequencing the 5'-32P-labeled RNA. The sequence is NpppGUUUUUAUUUUUAAUUUUCUUUCAAAUACUUCCAUCAUGAGUUCUUCACAAAAGAAAGCUGGUGGGAAAGCUGG. The AUG initiator codon is located 36 nucleotides in from the 5' end; the nucleotide sequence beyond corresponds to the amino acid sequence of the coat protein. This 5' noncoding region is rich in U (58% U); except for the 5'-terminal G, the next G in is part of the initiator AUG codon.


New method for localizing proteins in periodic structures: Fab fragment labeling combined with image processing of electron micrographs. Fab fragments prepared from antisera directed against purified bacteriophage T4 structural proteins and head-related structures were used to label proteins on the surface of T-even giant phage capsids. Optically filtered electron micrographs of the Fab-labeled capsids reveal both the location of specific proteins within the capsomeres and differing conformational states of the protein subunits. We describe parameters affecting the utility of this technique for the study of molecular organization and protein conformation in periodic biological structures.


Muscarinic acetylcholine receptors of the developing retina. Six- and 13-day chicken embryo retinas contain 10 and 320 fmol per mg of protein of specific binding sites for 3-[3H]quinuclidinyl benzilate, a ligand of muscarinic acetylcholine receptors. Most of the receptors of 13-day embryo retina were found, by autoradiography, to be localized in two sharp bands within the inner synaptic layer of the retina. In the adult, the receptors were found almost exclusively in three bands in the inner synaptic layer of the retina. A possible mechanism for generating sets of stratified or columnar neurons and relating one set to another is proposed.


Histone H3 disulfide dimers and nucleosome structure. The arginine-rich histone, H3, isolated from avian erythrocytes, can dimerize by forming a disulfide linkage between the single cysteine sulfhydryl residues at position 110 of the H3 polypeptide chain. The H3 dimer can be substituted for undimerized H3 in experiments in which the nucleosome is reconstituted from DNA and mixtures of the four "core" histones, H2A, H2B, H3, and H4. We report here that reconstituted nucleosomes containing H3 dimer are indistinguishable, by a number of criteria, either from native nucleosomes or from reconstitutes containing H3 monomer. The criteria include the pattern of susceptibility of the complex to nucleases, the amount of DNA supercoiling induced by histone binding, and the hydrodynamic properties of reconstituted nucleosome "core" preparations. The results suggest that the residues in the neighborhood of position 110 on each H3 molecule are in close contact in the nucleosome. If, as has been proposed, the nucleosome has a dyad axis, then the disulfide bridge between H3 molecules must lie on this axis.


Reversible and irreversible inhibition by anesthetics of the calcium-induced luminescence of aequorin. The actions of various kinds of so-called membrane-stabilizing drugs on Ca2+-induced flash intensity of purified aequorin, a photoprotein obtained from Aequorea aequorea, were examined. All drugs used in this experiment, including inhalational anesthetics, tetracaine, chlorpromazine, and morphine, depressed flash intensity, and the inhibition increased with time. The inhibition followed a mixture of reversible and irreversible kinetics. A method of analyzing irreversible kinetics is presented. The dissociation constants for reversible inhibition and the rate constants for irreversible inhibition are presented for each agent.


Electron microscopy of fibers and discs of hemoglobin S having sixfold symmetry. Aggregated forms of deoxyhemoglobin S were examined with a field emission transmission electron microscope. Images of isolated helical fibers were obtained from sickled cell lysates stained directly on the electron microscope grid. Optical and digital analyses of the electron micrographs showed that the fibers are similar to those characterized by J. T. Finch, M. F. Perutz, J. F. Bertles, and J. Döbler [(1973) Proc. Natl. Acad. Sci. USA 70, 718-722] in that they consist of stacked discs each composed of six hemoglobin molecules. The fibers exhibit an outer diameter of 160-170 A and an inner diameter of about 60 A with an axial spacing of 58 A per disc. The fiber can be described as a helix consisting of 56 discs per helical turn. We observed discs of six hemoglobin molecules, which may be stable substructural components of the fibers. They were observed in preparations of hemoglobin fibers and exhibited 6-fold symmetry by power spectrum analysis. A reconstructed image of a disc digitally filtered for 6-fold symmetry has a maximum external diameter of approximately 170 A and a central hole of 60 A diameter and is similar to the axial projection of a single disc from a low-resolution, three-dimensional reconstructed model of a fiber.


Random walk analysis of potassium fluxes associated with nerve impulses. We present a novel recursion method for obtaining theoretical expressions for unidirectional single-file fluxes of ions through narrow membrane channels containing an arbitrary number of ion sites. The theory is applied to experimental tracer fluxes associated with nerve impulses from cephalopod giant axon membranes at various temperatures between 7 degrees and 27 degrees. The comparison between the theoretical and experimental one-way fluxes suggests that the potassium channel in nerve membrane contains three ion sites, which is consistent with the deduction by Hodgkin and Keynes that the potassium channel contains two or three sites on the basis of the ratio of tracer influx to tracer efflux. The analytical results in this paper provide a further test of the single-file model for nerve and other membrane preparations.


Genetic dissection of the control of normal differentiation in myeloid leukemic cells. Normal myeloid precursors and MGI(+)D(+) myeloid leukemic cells can be induced to differentiate to mature cells by the normal protein inducer MGI. The sequence of differentiation is the induction of C3 and Fc rosettes, C3 and Fc immune phagocytosis (IP), synthesis and secretion of lysozyme, and formation of mature macrophages and granulocytes. Mutant clones of myeloid leukemic cells have been isolated with differences in the time of induction of C3 and Fc rosettes and C3 and Fc IP, in which lysozyme was induced without going through the stage of Fc or C3 IP, and with differences in inducibility by MGI to mature macrophages or granulocytes. Only one out of five MGI(-)D(-) clones gave rise to MGI(+)D(+) mutants. The ability to obtain mutants from this clone was associated with its special chromosome constitution, and these mutants showed a change in their ability for cap formation by concanavalin A. The steroid inducer dexamethasone can induce in MGI(+)D(+) clones differentiation to macrophages but not to granulocytes. Differentiation by steroid inducer in different clones occurred either with or without induction of Fc rosettes and Fc IP, and induction of C3 rosettes was not always associated with induction of C3 IP. The use of mutants that differ in their competence to be induced by MGI or steroid inducer has shown that there are separate controls for the induction of C3 and Fc rosettes, C3 and Fc IP, lysozyme, macrophages, and granulocytes.


Nerve growth factor prevents vinblastine destructive effects on sympathetic ganglia in newborn mice. Vinblastine injections in newborn mice produce severe atrophy of sympathetic ganglia; after a 7-day treatment the ganglia are 80% reduced in volume. Histological examinations show that this effect is due to a marked decrease in the neuronal cell population. The most precocious ultra-structural alterations are localized in the nuclear compartment, followed by axonal swelling and microtubule disappearance. Simultaneous injections of nerve growth factor entirely prevent the noxious effects of the vinca alkaloid, and result in partial appearance of the growth effects of nerve growth factor. Such protective action is not due to inhibition of vinblastine uptake which is the same in control mice and in mice pretreated with nerve growth factor. It is suggested that nerve growth factor prevents the vinca alkaloid action by favoring the assembly or organization, or both, of microtubules, which, from in vitro studies, have been proved to be inaccessible to vinblastine.


Mosaic mice with teratocarcinoma-derived mutant cells deficient in hypoxanthine phosphoribosyltransferase. Mutagenized stem cells of a cultured mouse teratocarcinoma cell line were selected for resistance to the purine base analog 6-thioguanine. Cells of a resistant clone were completely deficient in activity of the enzyme hypoxanthine phosphoribosyltransferase (HPRT, IMP:pyrophosphate phosphoribosyltransferase, EC 2.4.2.8), the same X-linked lesion as occurs in human Lesch-Nyhan disease. After microinjection into blastocysts of another genetic strain, the previously malignant cells successfully participated in normal embryogenesis and tumor-free, viable mosaic mice were obtained. Cells of tumor lineage were identified by strain markers in virtually all tissues of some individuals. Mature function of those cells was evident from their tissue-specific products (e.g., melanins, liver proteins). These mutagenized teratocarcinoma cells are therefore developmentally totipotent. Retention of the severe HPRT deficiency in the differentiated state was documented in extracts of mosaic tissues by depressed specific activity of the enzyme, and also by presence of unlabeled clones in autoradiographs of explanted cells incubated in [(3)H]hypoxanthine. Some mosaic individuals had mutant-strain cells in only one or a few tissues. Such animals may provide unique opportunities to identify the tissue sources of particular aspects of the complex disease syndrome. The tissue distribution of HPRT-deficient cells suggests that selection against them is particularly strong in blood of the mosaic mice, as is already known to be the case in human heterozygotes. This phenotypic parallelism supports the expectation that afflicted F(1) male mice that might be obtained from mutant germ cells can serve as a model of the human disease.


Angiotensin stimulation of bovine adrenocortical cell growth. Factors controlling proliferation of adrenocortical cells have been studied in monolayer cultures of bovine adrenocortical cells. Angiotensin II stimulated cell proliferation and [3H]thymidine incorporation into DNA with a half-maximal effective concentration of 0.96 +/- 0.27 nM. Similar sensitivity to angiotensin III with reduced sensitivity to angiotensin I and tetradecapeptide renin substrate was observed. Although sensitivity to angiotensin II was equivalent to that for fibroblast growth factor (1.5 nM half-maximal effective concentration), maximal effects of angiotensin were less than for fibroblast growth factor and serum. High concentrations of insulin (1-10 micrometer) also stimulated [3H]thymidine incorporation into DNA and cell proliferation. [Sar1,Ile5,Ile8]Angiotensin II, a competitive antagonist of angiotensin II, blocked angiotensin II stimulation of DNA synthesis but did not affect fibroblast growth factor and insulin stimulation of DNA synthesis. Corticotropin (ACTH) blocked the stimulatory effects of both angiotensin II and fibroblast growth factor. The dose-response curves for angiotensin II stimulation of steroidogenesis were similar to those for stimulation of [3H]thymidine incorporation into DNA. Among the seven cell types examined, only adrenocortical cells responded to angiotension II with stimulation of DNA synthesis.


Evidence that pyrimidine dimers in DNA can give rise to tumors. The Amazon molly, Poecilia formosa, is a small fish that grows in clones. Hence, cells from one animal may be transplanted to another without danger of rejection. Cells from thyroid and adjacent tissue were irradiated with UV light in vitro and injected into the abdominal cavity of isogeneic recipients. At appropriate UV doses and numbers of cells injected, all recipients showed exuberant thyroid proliferation. We give arguments and data indicating that the proliferation is a tumor, not a goitrogenic response. If the UV irradiation is followed, but not preceded, by photoreactivating illumination, the yield of thyroid growths is markedly decreased. Because other investigations have shown that photoreactivation monomerizes UV-induced cyclobutylpyrimidine dimers in DNA and does not affect other photoproducts, our data indicate that pyrimidine dimers in DNA can give rise to tumors.


DNA degradation in terminally differentiating lens fiber cells from chick embryos. During the terminal differentiation of lens fiber cells, nuclear DNA is known to accumulate free 3'-OH ends and is progressively lost from the nucleus. Toward the end of this process, nuclei undergo pycnosis and disappear. The size of the DNA in the epithelia and in early and late stages of fiber cell development was examined by electrophoresis on nondenaturing agarose/polyacrylamide gels. Low molecular weight DNA of discrete sizes appears only at the final stages of nuclear degeneration in central fiber cells and persists after the disappearance of the nuclei. These low molecular weight DNA fragments appear as multiples of a monomeric unit and are similar to the fragments produced by the digestion of epithelial cell nuclei by micrococcal nuclease. The data indicate that in lens fiber nuclei the double-strand breaks in vivo affect the chromatin during nuclear degeneration, and the data suggest that the DNA of these cells is organized into chromatin composed of discrete subunits.


Transitions in collagen types during matrix-induced cartilage, bone, and bone marrow formation. The localization of types I, II, and III collagens during bone matrix-induced sequential differentiation of cartilage, bone, and bone marrow was studied by specific immunofluorescence. Subcutaneous transplantation of coarse powders of demineralized rat bone matrix into allogeneic recipients resulted in new bone formation. After a transient appearance of polymorphonuclear leukocytes in the implant, fibroblasts appeared in close continguity to the matrix on day 3. Type III collagen was then localized as a fine network around the invading fibroblasts. On days 4--6 smaller amounts of type I were also detected around these proliferating cells. With the onset of chondrogenesis, type II collagen was detected in the cartilage matrix on day 6 and persisted until the early stages of bone formation. Vascular invasion of the implant was accompanied by osteogenesis on day 10. Type I collagen was demonstrated in the newly deposited bone matrix coating the surfaces of cartilage spicules and particles of implanted bone powder. On day 17 and thereafter, type III collagen was localized as a fibrous array around nests of hematopoietic cells.


Post-translational cleavage of presecretory proteins with an extract of rough microsomes from dog pancreas containing signal peptidase activity. The protease(s) responsible for removing the amino-terminal extension of nascent presecretory proteins (signal peptidase) has been extracted from rough microsomes of dog pancreas with the detergent sodium deoxycholate. Preprolactin and pre-growth-hormone, prepared by in vitro translation of bovine pituitary RNA in the wheat germ system, were used to assay signal peptidase in the extract. When added to the wheat germ system during translation, the extract reduced the size of preprolactin and pre-growth-hormone to that of prolactin and growth hormone, respectively. Post-translational addition of the extract also reduced the size of preprolactin and pre-growth-hormone to that of the authentic hormones. The prolactin produced by post-translational cleavage of radiolabeled preprolactin has been shown, by partial amino-terminal sequence analysis, to have the correct amino terminus. This post-translational assay has permitted the investigation of the subcellular localization of the enzyme. Sodium deoxycholate extracts of rough microsomes were active, whereas extracts of smooth microsomes were inactive. However, without detergent treatment, neither rough nor smooth microsomes were capable of cleaving preprolactin in the post-translational assay. From this we conclude that the signal peptidase activity is confined to the rough endoplasmic reticulum and is latent. Finally, we have detected two small peptides which we believe could be the signal peptides generated by the endoproteolytic cleavage of preprolactin and pre-growth-hormone by signal peptidase.


Liposome--lymphocyte interaction: saturable sites for transfer and intracellular release of liposome contents. The water-soluble dye 6-carboxyfluorescein was trapped in the internal aqueous compartments of small sonicated dioleoyl lecithin vesicles and used to assess the kinetics of transfer of vesicle contents to human lymphocytes. By using flow microfluorometry, the initial rate of dye transfer to the cells was measured as a function of the concentration of vesicles in the external medium. The rate of transfer consists of at least two components, one of which saturates at high vesicle concentration and the other of which does not saturate in the range of concentrations explored. The saturable component was competitively inhibited by vesicles not containing dye. Both the saturable and nonsaturable components of transfer were inhibited by fetal calf serum or bovine serum albumin but neither component was affected by bovine IgG, choline chloride, or heparin. Pretreatment of the lymphocytes with trypsin or Pronase had no effect on either component. The saturable component can be interpreted in terms of a two-step process in which vesicles bind reversely to sites on the cell surface, and dye is then transferred into the cell from the vesicle-site complex.


Molecular evolution of human and rabbit beta-globin mRNAs. The primary structures of human and rabbit beta-globin mRNAs are compared. Using as a standard the extent of nucleotide substitutions inferred from the hypervariable amino acid residues of fibrinopeptides A and B, which are thought to change largely by neutral evolution, we show that not all silent mutations in globin mRNA are neutral. The divergence of the sequences is limited in part by the selective usage of synonymous codons. The divergent nucleotides tend to be distributed nonrandomly: in the coding region silent substitutions are most rare in segments that are also deficient in substitutions leading to replacements.


Assignment of the human gene for galactose-1-phosphate uridyltransferase to chromosome 9: studies with Chinese hamster-human somatic cell hybrids. Chinese hamster-human somatic cell hybrids were analyzed for the expression of human galactose-1-phosphate uridyltransferase (GALT; UDPglucose:alpha-D-galactose-1-phosphate uridyltransferase, EC 2.7.7.12) by electrophoresis and for the presence of human chromosomes cytogenetically with the aid of Q-banding. Three of the 10 randomly chosen independently derived primary hybrid lines showed the presence of human GALT. Human chromosome 9 was consistently present in the hybrid lines expressing human GALT and consistently absent in the lines not expressing it. Biochemical analysis alone of 11 independently derived hybrid lines showed human GALT to be syntenic with known chromosome 9 markers (soluble aconitase, adenylate kinase 1, and adenylate kinase 3). Previous studies on chromosome assignment of this locus, utilizing somatic cell hybrids, have yielded inconsistent results; one group assigned GALT to chromosome 2, and another assigned it to chromosome 3. However, we believe that, based on our results and other published evidence, the correct assignment of the human GALT locus is to chromosome 9.


Increased dexamethasone resistance of cystic fibrosis fibroblasts. We demonstrate previously that fibroblasts from cystic fibrosis (CF) patients are significantly more resistant to the toxic effects of ouabain than normal human fibroblasts are. Ouabain is generally assumed to act primarily at the level of the cell membrane and to cause killing of cells by inhibiting ion transport. We have therefore examined the ability of normal and CF cells to survive exposure to ethacrynic acid, another inhibitor of ion transport, and to survive colchicine and aminopterin, resistance to which has been associated with membrane alterations in other cells. The effect of dexamethasone, which has a sterol nucleus similar to that of ouabain but is thought to have a different site of cellular action, was also tested. Exposure of the cells to ethacrynic acid, colchicine, or aminopterin did not reveal any differences in survival between normal and CF fibroblasts. However, CF cells survived exposure to dexamethasone significantly better than normal cells did. These results suggest that normal and CF cells do not differ in terms of a generalized resistance to ion transport inhibitors or to drugs that must pass through the membrane to be active. Instead, the results raise the possibility that CF cells have an enhanced resistance to drugs that have the sterol nucleus found in ouabain and dexamethasone.


Microsurgically produced homozygous-diploid uniparental mice. Shortly after fertilization, either the male or the female pronucleus was microsurgically removed from 202 F(1) hybrid eggs derived from crosses of two inbred strains. Subsequent incubation of these haploid eggs in medium containing cytochalasin B, which inhibits cytokinesis but not nuclear division, enabled the remaining pronucleus to become diploid. After nuclear diploidization and transfer to regular culture medium, cleavage commenced normally, and a total of 135 successfully manipulated eggs continued in development and yielded 93 morulae and blastocysts. These embryos were surgically transferred to the uteri of pseudopregnant foster mothers who gave birth to seven live female offspring. Five of the females were derived from the maternal genome (gynogenesis) and the remaining two mice inherited only the paternal genes (androgenesis), depending on whether the female or male pronucleus had been retained in the egg, respectively. Homozygosity for a number of genetic loci positioned on different chromosomes and effecting the coat color phenotype and strain-specific allelic variants of several enzymes, urinary and plasma proteins, and hemoglobins could be demonstrated unequivocally in all instances. Chromosomal analysis revealed a normal diploid karyotype including two X chromosomes. Thus far, six of the seven homozygous-diploid (isogeneic) females have proved to be fertile and have given birth to progeny corresponding only to the pronuclear genotype of the mother.


Design of molecular control mechanisms and the demand for gene expression. Regulation by a repressor protein is the mechanism selected when, in the organism's natural environment, there is low demand for expression of the regulated structural genes. Regulation by an activator protein is selected when there is high demand for expression of the regulated structural genes. These general conclusions are useful in relating physiological function to underlying molecular determinants in a wide variety of systems that includes repressible biosynthetic pathways, inducible biosynthetic enzymes, inducible drug resistance, and prophage induction, as well as inducible catabolic pathways, for which a special case of this prediction previously was reported [Savageau, M. A. (1974) Proc. Natl. Acad. Sci. USA 71, 2453-2455].


Antigen-binding mutants of mouse myeloma cells. A cultured mouse myeloma cell line, S107, that secretes an IgA phosphocholine-binding immunoglobulin has been cloned in soft agar and overlaid with phosphocholine-hemocyanin. Spontaneous mutants that secrete immunoglobulin with a decreased ability to precipitate antigen were detected with this plate assay and occur at a very high frequency. From one such mutant, phenotypic revertants arise spontaneously with a frequency of 0.28-2.8%. This mutant and one of its revertants were studied, and they were found to differ from the parent and from each other serologically and in antigen binding. While it is not yet clear whether these findings bear any relationship to the normal generation of antibody diversity, they do indicate that it is possible to generate antigen binding diversity in somatic cells.


Interactions of C-reactive protein and complement with liposomes. Interactions between C-reactive protein (CRP) and liposomal model membranes containing phosphatidylcholine were investigated. These interactions, in the presence of human serum, resulted in consumption of each of the components of the classical complement pathway (C1-C9) and also resulted in complement-dependent damage and release of trapped glucose from certain types of liposomes. CRP-initiated lysis of liposomes was strongly dependent upon membrane lipid composition. Optimal activity occurred with positively charged liposomes containing galactosylceramide (galactocerebroside); positively charged liposomes lacking galactocerebroside released much less glucose, while negatively charged liposomes, either with or without galactocerebroside, did not release glucose at all. Glucose release was inhibited by free phosphocholine. Lesser, but significant, "background" glucose release independent of the presence of CRP also was observed with positively charged liposomes containing galactocerebroside, and this was associated with marked preferential consumption of the later-acting complement components (C3-C9). C2-deficient human serum failed to support CRP-dependent glucose release, but glucose release was observed upon reconstitution of the serum with C2. Guinea pig complement also did not support CRP-mediated glucose release, but upon addition of human C1q substantial glucose release was observed. We conclude that (i) CRP can sensitize appropriate liposomes for complement-dependent damage via the primary complement pathway starting at the level of C1q; (ii) of those studied, liposomes that are most susceptible to membrane damage contain phosphatidylcholine, have a positive charge, and contain a ceramide glycolipid; and (iii) such liposomes also are sensitive, although to a much lesser degree, to complement-dependent lysis initiated in the absence of CRP and involving consumption of terminal in excess of early acting complement components.


Evidence for ovarian "inhibin": suppression of the secondary rise in serum follicle stimulating hormone levels in proestrous rats by injection of porcine follicular fluid. The present studies were carried out to see if porcine follicular fluid could inhibit increases in serum follicle stimulating hormone (FSH) levels when injected into the rat. For these studies the pentobarbital-treated proestrous rat was chosen as the major test animal model. If an artificial surge of luteinizing hormone (LH) is administered to these rats, it can induce a synchronized secondary rise in FSH secretion rate. Normal saline-treated rats were also used as test animals. They exhibit preovulatory endogenous "surges" of LH and FSH, and also a secondary FSH rise. Porcine follicular fluid was harvested from medium-sized and large (3- to 10-mm diameter) follicles and treated with charcoal to remove endogenous steroids. Charcoal-treated porcine serum served as a control solution. The fluid was injected intraperitoneally in two 0.5-ml doses into pentobarbital-treated proestrous rats immediately and 3 hr after LH injection. Follicular fluid, but not the serum, suppressed the secondary, LH-induced FSH rise (P < 0.01) in a dose-dependent manner, without altering the effects of LH upon serum ovarian steroid levels or follicular rupture. It was effective down to a total dose of 200 mul. Porcine follicular fluid also blocked the secondary FSH surge in normal proestrous rats exhibiting endogenous LH/FSH primary surges. Thus, it would appear that porcine follicular fluid contains a non-steroidal substance(s) that can block the secretion of FSH that is secondary to a natural or artificial LH surge.


Demonstration of different contractile mechanisms for angiotensin II and des-Asp1-angiotensin II in rabbit aortic strips. Evidence of selective inhibition, differences in dose-response relationships, and cross-tachyphylaxis studies suggest that separate receptors and/or mechanisms may be involved in responses to angiotensin (Ang), [Sar1]Ang II, and Ang III (= des-Asp1-Ang II). The extracellular Ca2+ requirement for contractile responses induced by angiotensin peptides and norepinephrine was determined in rabbit aortic strips. Responses to K+ and [Sar1]Ang II were attenuated markedly by treatment with SKF-525A, verapamil, or Ca2+-free buffer. The response to Ang II was not impaired by verapamil, was blocked partially by SKF-525A, and was reduced markedly in Ca2+-free medium. Norepinephrine- and Ang III-induced contractions were not dependent on extracellular Ca2+. K+, Ang II, and [Sar1]Ang II required extracellular Ca2+ to induce contraction of the rabbit aorta. The data indicate that Ang III may have a mechanism of action that differs from that of [Sar1]Ang II and Ang II.


Mapping of human chromosomal regions related to neoplasia: evidence from chromosomes 1 and 17. In clonal aberrations leading to an excess or partial excess of chromosome 1, trisomy for bands 1q25-1q32 was noted in the myeloid cells from all of 34 patients who had various disorders such as acute leukemia, polycythemia vera, and myelofibrosis. This was not the result of a particularly fragile site in that region of the chromosome because the break points in reciprocal translocations that involve it occurred almost exclusively in the short arm. Two consistent rearrangements that have been observed in chromosome 17 produced either duplication of the entire long arm or a translocation of the distal portion of the long arm to chromosome 15. The nonrandom chromosomal changes found in hematologic disorders can now be correlated with the gene loci on these chromosomes or chromosomal segments. Seventy-five genes related to various metabolic enzymes have been mapped; it may be significant that chromosomes carrying gene loci related to nucleic acid metabolism are more frequently involved in hematologic disorders (and other malignancies as well) than are gene loci related to intermediary or carbohydrate metabolism. Furthermore, the known virus-human chromosome associations are closely correlated with the chromosomes affected in hematologic disorders. If one of the effects of carcinogens (including viruses) is to activate genes that regulate host cell DNA synthesis, and if translocations or duplications of specific chromosomal segments produce the same effect, then either of these mechanisms might provide the affected cell with a proliferative advantage.


Molecular basis of reovirus virulence: role of the S1 gene. A genetic approach has been used to define the molecular basis for the different patterns of virulence and central nervous system cell tropism exhibited by reovirus types 1 and 3. Intracerebral inoculation of reovirus type 3 into newborn mice causes a necrotizing encephalitis (without ependymal damage) that is uniformly fatal. Animal inoculated with reovirus type 1 generally survive and may develop epedymal cell damage (without neuronal necrosis) and hydrocephalus. Using recombinant clones derived from crosses between reovirus types 1 and 3, we have been able to determine that the S1 genome segment is responsible for the differing cell tropism of reovirus serotypes and is the major determinant of neurovirulence. The type 1 S1 genome segment is responsible for ependymal damage with subsequent hydrocephalus; the type 3 S1 genome segment is responsible for neuronal necrosis and neurovirulence. We postulate that these differences are due to the specific interaction of the sigma1 outer capsid polypeptide (the protein coded for by the S1 genome segment) with receptors on the surface of either ependymal cells or neuronal cells.


Histrionicotoxin enhances agonist-induced desensitization of acetylcholine receptor. Dihydroisohistrionicotoxin inhibits acetylcholine receptor-dependent 22Na+ uptake of cultured chick muscle cells with a KI of 0.2 micrometer. The inhibition is noncompetitive with respect to agonists. The toxin enhances desensitization of the receptor by agonists which is accompanied by a 10-fold increase in receptor affinity for agonists. Dihydroisohistrionicotoxin increases the affinity of the desensitized form of the receptor for agonists but not antagonists. The results suggest that dihydroisohistrionicotoxin inhibits the acetylcholine receptor by causing an increase in the affinity of the desensitized form of the receptor for agonists and thereby stabilizing the desensitized state.


Effect of increased potassium concentrations on particle motion within a neurosecretory structure. Intensity fluctuation spectroscopy was used to detect particle motion within the pericardial organ of the crab Carcinus while it was perfused with media with increased potassium concentrations. Replacement of some or all of the sodium ions of the normal medium with potassium ions caused a graded increase in motion. Evidence is presented which indicates that this effect is not a consequence of membrane depolarization or calcium ion influx, but is due to the flux of potassium chloride and water into the cytoplasm of the cells.


Determination of transmitter function by neuronal activity. The role of neuronal activity in the determination of transmitter function was studied in cultures of dissociated sympathetic neurons from newborn rat superior cervical ganglia. Cholinergic and adrenergic differentiation were assayed by incubating the cultures with radioactive choline and tyrosine and determining the rate of synthesis and accumulation of labelled acetylcholine and catecholamines. As in previous studies, pure neuronal cultures grown in control medium displayed much lower ratios of acetylcholine synthesis to catecholamine synthesis than did sister cultures grown in medium previously conditioned by incubation on appropriate nonneuronal cells (conditioned medium). However, here we report that neurons treated with the depolarizing agents elevated K(+) or veratridine, or stimulated directly with electrical current, either before or during application of conditioned medium, displayed up to 300-fold lower acetylcholine/catecholamine ratios than they would have without depolarization, and thus remained primarily adrenergic. Elevated K(+) and veratridine produced this effect on cholinergic differentiation without significantly altering neuronal survival. Because depolarization causes Ca(2+) entry in a number of cell types, the effects of several Ca(2+) agonists and antagonists were investigated. In the presence of the Ca(2+) antagonists D600 or Mg(2+), K(+) did not prevent the induction of cholinergic properties by conditioned medium. Thus depolarization, either steady or accompanying activity, is one of the factors determining whether cultured sympathetic neurons become adrenergic or cholinergic, and this effect may be mediated by Ca(2+).


Costs of maintenance and growth in genetically obese (obob) mice.


Androgens and long-term complete remission in acute granulopoietic leukemias.


Humanism and Medicine.

Humanism and Medicine.


Rehabilitation of the knee following surgery or injury.


Dietary goals for the United States: statement of The American Medical Association to the Select Committee on Nutrition and Human Needs, United States Senate.


Health perspectives in Rhode Island: a sample survey.


Distribution of 59Fe-labelled iron-poly(soribitol-gluconic acid) complex in normal and anaemic rats. An iron carbohydrate complex, iron-poly(sorbitol-gluconic acid), Ferastral, was labelled with 59Fe, and its distribution in rats was studied. The animals were intramuscularly treated with a dose of 10 mg of iron/kg. Three groups of animals were used: group A: non-anaemic and group B: anaemic rats, both kept on iron-deficient diet, and group C: non-anaemic rats kept on iron-supplemented diet. Urinary and faecal excretion, distribution in the body and incorporation in blood of the 59Fe was followed up to 28 days. The total excretion after that time was 15%. There was a rapid initial phase followed by a slower continuous one. After 28 days group A had 25, group B 13 and group C 40% of the given dose remaining at the site of injection. The corresponding values in liver after 28 days were 7, 4 and 17% of the given dose, respectively. In blood a continuous increase was observed. At 28 days after administration 26, 43 and 17% of the given dose had been incorporated in the red blood corpuscles of the respective groups. These results show that the iron complex is absorbed from the site of injection and is utilized for haemoglobin synthesis. They also show that the disposition of the complex is influenced by the iron content of the diet.


The absorption and availability of iron from iron-poly(sorbitol-gluconic acid) complex in rats as measured by phlebotomy. The local and systemic availability of the iron in an iron-poly(sorbitol-gluconic acid) complex (Ferastral) has been studied in rats. The animals were treated intramuscularly with the complex in a dose corresponding to 10 mg of iron per animal. The technique of repeated phlebotomy was used to induce and maintain constant levels of anaemia. The phlebotomies were started immediately after injection and at 14 and 28 days after dosing. At the end of the appropriate interval, the remaining amount of iron at the injection sites as well as the degree of utilization of the iron retained in the animals were estimated. It was shown that the amount of iron at the site of injection of these anaemic animals was less than 3% regardless of the time when the series of phlebotomies were started. It was also shown that the degree of utilization of the iron from the complex was 90% of the retained amount of iron when the phlebotomies immediately followed dosing, and 86% and 79% when initiated 14 days and 28 days respectively. The significance of the decrease in utilization is discussed.


Disposition in rats of the carbohydrate polymer in Ferastral--an iron-poly (sorbitol-gluconic acid) complex. The disposition and pharmacokinetics in rats of 14C-sorbitol-Ferastral has been studied as an initial step towards elucidating the metabolic fate of the polymeric organic moiety. The period of time studied was 24 h. Following i.v. administration of a dose corresponding to 10 mg/kg of iron approximately 60% of the isotope was found in the urine after this period of time. A further 3% was demonstrated in the faeces whilst the expired air accounted for about 10% in the form of 14CO2. Thus, a relatively large proportion of the radiolabel was promptly excreted via the kidneys whilst the expired air and probably bile constituted secondary excretion routes. The physico-chemical nature of the urinary product was similar to the parent carbohydrate compound. The residual 14C radioactivity was distributed mainly between the liver, which contained approximately 45% of the retained label, carcass with 36% and large intestine with 7%. The plasma disappearance pattern was biphasic. Similar experiments with the 14C-sorbitol polymer starting material were performed for purposes of comparison.


Distribution of the carbohydrate polymer in 14C-labelled iron-poly (sorbitol-gluconic acid) complex after intramuscular administration in normal and anaemic rats. The carbohydrate polymer in an iron-poly (sorbitol-gluconic acid) complex was prepared by using uniformly labelled 14C-sorbitol or 14C-gluconic acid. The distribution of these complexes 7, 14 and 28 days after administration has been followed in non-anaemic and anaemic rats. Most of the 14C from the labelled complexes was found in the expired CO2, faeces and urine. Twenty-eight days after injection of 14C-labelled Ferastral, less than 13% of the given dose of either 14C-sorbitol or 14C-gluconic acid labelled preparation could be found in the body. There was no specific accumulation of 14C in any organ except in the bones, where 3% of the given dose could be seen 28 days after 14C-gluconic acid labelled Ferastral to both non-anaemic and anaemic animals.


Studies on the placental transfer in rats of 59Fe- and 14C-labelled iron-poly (sorbitol-gluconic acid) complex. The placental transfer of an iron-poly (sorbitol-gluconic acid) complex, Ferastral, has been studied. By use of the 59Fe-, 14C-sorbitol, and 14C-gluconic acid labelled complexes it was shown that the passage of the iron moiety is related to foetal need and that only negligible amounts of 14C from either 14C-sorbitol or 14C-gluconic acid labelled iron complexes were found in the foetus. This implies that the intact complex does not pass the placental barrier.


Metabolic studies of iron-poly (sorbitol-gluconic acid) complex, Ferastral. The metabolism of iron-poly (sorbitol-gluconic acid) complex (Ferastral) was studied in 6 blood donors with latent iron deficiency and 13 patients with manifest iron deficiency anaemia. Between 125 and 1 000 mg of iron were injected. In latent deficiency an average of 19% of the dose given was excreted during the first two weeks. The corresponding figure for patients with manifest iron deficiency was 11%. An average of 8% of the dose was retained at the site of injection after 40 days. In the two groups, respectively, 61% (range 47%-70%) and 58% (range 24%-86%) of the iron retained was incorporated into red cells after 40 days, which is judged to be quite satisfactory. Some local side effects were observed.


Studies on iron stores built up by an iron-poly (sorbitol-gluconic acid) complex, Ferastral, in man. Preliminary report. The characteristics of iron stores built up by a new parenteral iron-poly (sorbitol-gluconic acid) complex, Ferastral, have been studied in iron depleted non-anemic blood donors. The results from studies on the first three subjects are presented. The availability of this storage iron for the chelator, desferrioxamine, was found to be in the same range as normal iron stores. The pattern of distribution in reticulo-endothelial cells of the bone marrow could not be differentiated from natural storage iron. No visible iron could be detected in liver parenchymal cells 40 days after iron administration. The results from these preliminary studies suggests normal bioavailability of this material for Hb-synthesis. The absence of iron in liver parenchymal cells might be explained by the short time interval between the iron administration and the fine needle aspiration biopsy.


Absorption, excretion, and utilization of an intramuscular iron-poly (sorbitol-gluconic acid) complex, Ferastral. An iron-poly (sorbitol-gluconic acid) complex (Ferastral) labelled with 59Fe was administered by intramuscular injection to clinically and haematologically normal male volunteers. Urinary excretion of radioactivity was measured for the first 48 hours following injection, and was 2%-10% of the administered dose. Disappearance of radioactivity at the site of injection and accumulation of radioactivity in the liver, spleen, and sacrum were measured by external scintillation counting. Plasma radioactivity was measured at intervals for four days and red cell incorporation of 59Fe reached levels of 29%-57.5%. An unexpected finding was that the highest levels of urinary excretion of radioactivity did not coincide with the highest levels of plasma radioactivity.


Interaction between Ferastral and plasma factors. Ferastral, iron-poly (sorbitol-gluconic acid) complex, in aqueous solution is completely adsorbed by magnesium carbonate; Ferastral in serum solution is only partially adsorbed by magnesium carbonate. Adsorption by magnesium carbonate removes the fraction of Ferastral in serum solution which donates iron more readily to transferrin. It is suggested that this fraction comprises the smaller molecular species of the Ferastral complex and that these species have a faster rate of absorption from an intramuscular injection site than the larger molecular species of the iron complex preparation. These findings shed some light on the patterns of serum transferrin bound iron compared with serum Ferastral concentrations which occur with time after intramuscular Ferastral injection.


The assay of iron-poly (sorbitol-gluconic acid) complex (Ferastral) and its separation from transferrin in serum. A new iron-poly (sorbitol-gluconic acid) complex (Ferastral) has been studied. A method of assay is described. The iron complex may be separated from serum transferrin using a Sephadex DEAE A50 column. This binds the iron complex and elutes iron-transferrin which can then be assayed. It is shown that the assay of serum transferrin unsaturated binding capacity using excess 59FeCl2 and MgCO3 adsorption, is valid in the presence of Ferastral. Serum unsaturated iron binding capacity may therefore be used to follow the binding of Ferastral iron by transferrin. These methods may be used to follow the distribution of iron in plasma after an intramuscular injection of Ferastral.


Studies on an iron-poly(sorbitol-gluconic acid) complex for parenteral treatment of iron deficiency anaemia. A preparation containing an iron-poly(sorbitol-gluconic acid) complex for parenteral treatment of iron deficiency anaemia is described. The physical and chemical properties of the iron complex have been studied by using electrophoresis and gel permeation chromatography. A rapid absorption from the injection site after intramuscular administration to rabbits takes place, 70% of the iron being absorbed after 24-48 hours. Thereafter, the absorption rate is slower, and 32 days after the injection 94% has been absorbed from the injection site. In rabbits the maximum level of iron in serum is reached after 12-24 hours; in dogs after 1-3 hours. Disappearance from the serum takes place slowly. The complex is exclusively absorbed via the lymphatic route. Nine to ten per cent of the given dose is excreted by the kidney within 72 hours in rats and 24 hours in rabbits after intramuscular administration. On administration of the preparation to rats, made anaemic by phlebotomy, a rapid increase of haemoglobin values is observed as well as a very high utilization of the retained amount of the given dose.


Transfer of iron from Ferastral and other organic complexes to transferrin as measured by reticulocyte uptake. The transfer of iron from the iron carbohydrate complexes, Ferastral, Imferon, and Jectofer, and from ferric chloride has been studied by the effect of such transfer in reducing reticulocyte uptake of 59Fe from labelled transferrin. There are plasma factors which augment the transfer of iron from complex to transferrin. The pattern of transfer from Ferastral and from Imferon are similar: at concentration of 5000 microgram/100 ml and 1250 microgram/100 ml in plasma these complexes transfer about 0.8% and 1.7%, respectively. Jectofer transfers about four times these amounts under similar conditions. In the case of Ferastral there is evidence of an equilibrium between transferrin-bound and Ferastral-bound iron. The characteristics of Ferastral assessed in this way suggest that it may prove suitable for therapeutic use as a total dose infusion.


Kinetic studies on an intramuscular iron-poly (sorbitol-gluconic acid) complex. Levels of iron and unsaturated iron binding capacity in serum and the renal excretion of iron were studied in 15 anaemic patients receiving Ferastral. Various dose schedules were used, 100 mg Fe daily for 10 days, 200 mg Fe daily for 5 days, and 500 mg Fe twice at various intervals from 2 to 7 days. A continuous increase of iron in serum was seen after the doses 100 mg and 200 mg daily as well as after 500 mg given at intervals of 2 and 3 days. Longer 500 mg dose intervals resulted in a fluctuation in the iron concentration. Peak levels varied between 740 and 5 260 microgram/100 ml (132 and 942 mumol/1). A transient decrease in the UIBC was seen. Urinary excretion of iron varied between 8.9 and 26.9% of the dose given. No local or general side effects occurred.


Patterns of plasma iron fractions after intramuscular Ferastral. The distribution of plasma iron fractions after intramuscular Ferastral is described for times up to eight days after injection. Transferrin-bound iron reaches a peak value in 4-8 hours and falls towards normal values while circulating iron-complex concentration remains high. The peak concentration of iron-complex occurs 8-24 hours after injection, and is virtually cleared from the circulation in 6-8 days. An approximate T 1/2 of 30 hours for plasma Ferastral is deduced from the rate of fall from peak values. UIBC values fell as transferrin-bound iron increased, but did not reach zero in any subject studied.


Compensation for blood loss in plastic surgery with ferastral and autologous blood transfusion. A study has been made to assess the effect on haematological status of pre-operative parenteral iron in a operation accompanied by conisderable blood loss. Forty-two patients who were to undergo breast reducing operations were selected for the trial. Patients were divided into three groups: group 1 which received no iron treatment, group 2 which received Ferastral, 250 mg Fe, 21 days before operation and group 3 which received Ferastral, 250 mg Fe, 21 days before operation plus an autologous blood transfusion during operation. The blood loss at operation which was estimated to be between 500 and 1 000 ml in 86% of the cases resulted in a decrease in haemoglobin in all groups. On the 14th postoperative day the decrease in Hb was significantly lower in both group 2 and 3 compared to group 1 (P less 0.05 and P less than 0.01, respectively). Forty per cent of the patients were anaemic in group 1, 10% in group 2 and 15% in group 3. It is concluded that the need for blood transfusion may be avoided in operations where the blood loss is less than 1 000 ml, provided sufficient iron is available for spontaneous blood replacement to take place.


Iron-deficiency in the tropics.

Iron-deficiency in the tropics. Different populations in the tropics show wide variation in their iron status, even when living in the same area. These differences range from the most profound iron-deficiency anaemia to nutritional haemosiderosis. The extremes of dietary iron content so far reported are 5 mg per day in Mauritius to 180 mg per day in Ethiopia. Contamination with inorganic iron is an important source in some communities. Generally, the iron in food is in excess of requirement, but lumenal factors are all important in deciding how much is available for absorption. The iron of some diets is readily absorbed, but in other only a small percentage is utilizable. Hookworm infection is the commonest cause of severe iron-deficiency in many parts of the world. Its prevalence and its density depends on social habits, and may vary considerably even within tribes. The development of iron-deficiency anaemia depends on the hookworm load being sufficient to cause iron loss in excess of available intake. Anaemia impairs work capacity, reproductive ability of women and the development of children, and has wide social and economic repercussions. An intramuscular preparation by which 500 mg of iron may be administered in one injection has most useful application in areas where medical care is minimal.


Pharmacological studies on an iron-poly-(sorbitol-gluconic acid) complex for parenteral treatment of iron deficiency anaemia. Intravenous injection of the iron-poly(sorbitol-gluconic acid) complex (IPSG) to cats anaesthetized with sodium pentobarbitone caused a transient decrease in mean arterial blood pressure and a temporary increase in central venous pressure, heart rate and femoral blood flow at large doses (cumulative doses up to 744 mg/kg). Tachyphylaxis developed upon repeated administration. A temporary reduction in the magnitude of the blood pressure responses to noradrenaline and isoprenaline was obtained after large doses of IPSG. The blood pressure effects of acetylcholine, histamine and bilateral carotid occlusion were not affected. No definite effects were seen on the electrocardiograms. The transient cardiovascular effects were interpreted as being due to the presence of small amounts of ferrous iron in the preparation. IPSG did not significantly affect blood coagulation or platelet aggregation as judged from results of in vitro and in vivo experiments. Immunogenicity studies in rabbits and antibody analyses of sera from patients treated with IPSG failed to demonstrate any effect of IPSG on the immune system.


Toxicological studies on ferastral, an iron-poly-(sorbitol-gluconic acid) complex.


Teratological, peri- and postnatal studies on Ferastral, an iron-poly(sorbitol-gluconic acid) complex.


Oncological study in rats of Ferastral, an iron-poly-(sorbitol-gluconic acid) complex, after intramuscular administration. Four groups of Sprague-Dawley rats were given Ferastral, an iron-poly(sorbitol-gluconic acid) complex (IPSG) or Imferon, an iron-dextran complex, intramuscularly twice a week for 17 weeks. The experiment lasted for 95 weeks. Each compound was given to two groups, one low dose group and one high dose group. Depending on the body mass, the dose levels varied between 2.5 and 10 mg (25-50 mg/kg body mass) and between 5 and 20 mg (50-100 mg/kg body mass) of iron per rat, respectively. The mean total dose of iron per rat was calculated to be 235 and 495 mg, respectively. Another group of animals served as a control. From about the 30th experimental week onwards tumours developed at the intramuscular injection sites in the groups given Ferastral and Imferon. The tumours appeared to be sarcomas. In almost all the treated animals that lived longer than 30 weeks after the start of the experiment, sarcomas were present at the intramuscular injection sites. The sarcomas appeared earlier in the high dose groups than in the low dose groups and slightly earlier in the rats given Ferastral than in those given Imferon. No other pathological changes, including neoplasms, were considered to be related to the treatment.


Determination of small quantities of fluoride in some biological materials.


The antiplaque effects of a nonflavored and a flavored chlorhexidine gluconate rinsing solution. A flavored chlorhexidine gluconate rinsing solution was compared with a nonflavored solution. Both solutions inhibited plaque growth significantly (60% PlI, 40% SPI). The differences between the flavored and the nonflavored solution were not significant.


Gingivitis in Zurich schoolchildren. A reexamination after 20 years. In 1957 the periodontal health of 1350 Zurich schoolchildren was assessed. The same investigator (M.Z.) examined 1217 children in the same school district 18 years later. 909 children were considered for statistical analysis. Only 9.1% were found to be free from gingival inflammation. 32% of the papillary and marginal gingival areas showed inflammatory signs. Prevalence of gingivitis was similar in different socioeconomic groups and in children treated by general practitioners and school dental clinics respectively. Gingivitis prevalence was remarkably similar in 1957 and 1975. An increase in the intensity of inflammation was, however, statistically significant. Reconsideration and modification of the existing measures in preventive periodontology are indicated.


Treatment of periodontal pockets with tricalcium phosphate in man. A preliminary report. After oral hygiene instruction 29 infrabony pockets in 9 patients were treated with deep scaling and flap surgery. The pockets were filled with a biodegradable tricalcium phosphate implant paste. 6 months postoperatively the average reduction of clinical pockets was 2.6 mm and the radiographically detectable attachment was 1.1 mm more coronal than preoperatively. 10% of the pockets remained deeper than 3 mm. Easy handling and simple storage are the predominant advantages of the described implantation material. Before recommending it for general use, however, some technical problems will have to be solved.


Endosteal oral implantology: two-year results of mobility and hysteresis measurements; A human study. Clinical experience with tooth mobility measurements for assessment of periodontal health was employed in evaluating endosteal implant status. Mobility and hysteresis measurements of 14 implants in 11 patients were carried out regularly during a 2-year period. Immediate postsurgical implant post displacement was 5 hundreths of a millimeter (5.10(-2) mm). After 1, 6, and 24 months, mobility had increased to 13, 21, and 54.10(-2) mm respectively. Viscoelastic recoil patterns were recognized as diagnostic indicators of profound changes in the biophysical properties of periimplant environment.


Dental caries 12 years after introduction of a preventive program-preliminary report from seven communities. Supervised toothbrushing with fluoride preparations and information on prevention of dental disease was introduced in 1963/64 for the majority of schools in the canton of Zurich. From 1963 to 1975, DMFT levels decreased by 47 to 54 percent, DMFS levels by 59 to 64 percent. Caries on surfaces of anterior teeth and on smooth surfaces of molars was reduced by 90 percent. The number of sound deciduous teeth increased substantially from 1963 to 1975.


Problem solving in diagnostics and therapeutics of neurology.


Athletic physicals. (A report for the South Dakota Chapter of the American Academy of Family Physicians).


Effect on speech of mandibular prognathism before and after surgical treatment.


The first 100 patients with jaw fractures at the Department of Oral Surgery, Dental School, Huddinge.


Caries progression in the first permanent molars. A longitudinal study.


In vitro corrosion of screwposts and amalgam placed in contact.


Selective coronary arteriography by a percutaneous femoral technique at West Virginia University Medical Center.


Appendicitis due to strangulation in a femoral hernia: report of a case.


Acute monocytic leukemia on stage IV neuroblastoma following radiation and cyclophosphamide therapy.


The aid of electron microscopy in the diagnosis of osteosarcoma of the bone (case report). Osteogenic sarcoma with marked cellular anaplasia and little or no osteoid formation may present major problems in histologic identification. The application of electron microscopy is demonstrated in a case of a poorly differentiated osteosarcoma of the tibia. Unequivocal electron microscopic diagnostic findings are described in relation to light microscopic data.


Levamisole as an immunotherapeutic agent in the treatment of cancer.


Transfer factor: a potential agent for immunotherapy of cancer.


Immunotherapy and chemoimmunotherapy of malignant disease with BCG and nonviable mycobacterial fractions.


The history of the American Society of Cytology.


The future of clinical cytology.

The future of clinical cytology.


The role and future of the pathologist in cytology.


A quarter of a century of cytology.

A quarter of a century of cytology.


The future of the American Society of Cytology.


Diagnostic and therapeutic aspects of cervical carcinoma. Based on the material of 991 cases.


Myometrial energy metabolism during pregnancy and normal and dysfunctional labor.


The future role of the cytotechnologist.


The role of the clinician in cytology and the Society: non-gynecologic cytology.


Development of osteosarcoma in rats after irradiation. The rat femur diaphysis was exposed to local continuous irradiation with various constant dose-rates for different periods of time and examined after different observation times. The incidence of osteogenic sarcomas per cm2 appeared to be comparable to that reported for animals exposed to internal irradiation by bone-seeking isotopes which involves systemic effects. The site of origin in 5 rats was found to be in or near the endosteum. Malignant proliferation started after at least 180 days of irradiation. Tumour development was dependent upon continued irradiation of an endosteum which contained a thin layer of fibroblasts and a few apparently inactive osteoblasts and osteoclasts. No increased cellular activity and no indication of new bone, cartilage or fibrosis were evident before the tumours began to develop.


Education and training of the pathologist in cytopathology.


Screening a hospital population for urinary tract cancer.


Presidential address.

Presidential address.


The interactions of smoking, environment and heredity and their implications for disease etiology. A report of epidemiological studies on the Swedish twin registries.


The incidence and significance of intracytoplasmic calcifications in nipple aspirate specimens. Intracytoplasmic deposits demonstrated to be calcifications have been identified in nipple aspirates from 3.6 per cent of breasts with satisfactory cytology. The deposits occurred almost exclusively in epithelial groups, and their structure ranged from dust-like particles and spiculated forms to psammoma bodies; all exhibited characteristic density, refractility and well-defined, irregular borders. Nipple aspirates containing calcifications were typically characterized by the presence of numerous epithelial groups, frequently of papillary type. This association was supported histologically by the fact that 11 of the 14 available tissue specimens contained papillary lesions in the duct system, including three with papillary carcinoma in situ. Cytologic abnormality was significantly associated with the presence of calcifications, and marked cellular changes were found four times as often in this group as in the total population with satisfactory cytology. Calcific deposits were found in six of the fourteen available tissue specimens, four of which contained breast carcinoma. Calcifications were noted in only 23 per cent of 97 available mammograms, but further investigation has revealed other significant radiologic findings. Cytologic calcifications were found most frequently in aspirates from women between the ages of 41 and 60 and were often associated with clinical findings of breast disease. Preliminary findings indicate an increased prevalence of breast carcinoma in patients with cytologic calcifications and suggest that such patients may require closer than normal follow-up.


Trichinosis in an immunosuppressed human host. The clinical presentation and autopsy findings in the case of a patient with acute myelomonocytic leukemia and trichinosis are described. Cellular immunity is an important host defense against nematode infections such as strongyloidiasis. It is therefore probable that the severe trichinosis seen in this immunosuppressed patient was more than coincidental. Due to concomitant failure of humoral immunity, serodiagnosis of trichinosis would have been impossible. The only means of making the diagnosis antemortem would have been muscle biopsy.


Ferruginous bodies in sputa of former asbestos workers. Routine cytopathologic examinations were performed at six-month intervals on sputum specimens from 628 former asbestos workers and 138 control patients. The occurrence of ferruginous bodies in sputa is found to increase as a logarithmic function of the length of occupational exposure to asbestos in workdays. No significant association is found between the occurrence of ferruginous bodies and the worker's age, smoking history, degree of cellular epithelial atypia, or time since last exposure. We conclude that the presence of ferruginous bodies in sputa is evidence of probable significant occupational exposure to asbestos dust. Their absence does not indicate lack of exposure. We can also conclude that routine cytopathology procedures are sufficient for the detection of ferruginous bodies in sputa.


The numerical composition of cellular samples from the female reproductive tract IV. Carcinoma in situ cases exhibiting other than normal vaginal flora. The numerical composition of vaginal, ectocervical and endocervical (VCE) cell samples from patients with carcinoma in situ of the uterine cervix exhibiting other than normal vaginal flora is described. The comparative results of cell counts in patients with carcinoma in situ with normal and other than normal vaginal flora are presented.


Chromosomes and causation of human cancer and leukemia: XXVIII. Value of detailed chromosome studies on large numbers of cells in CML. Comparison of the chromosome findings obtained on routine examination (10-50 cells) of the marrows from patients with Ph1-positive CML with those based on a large number (110-500 cells) of metaphases in six of these patients, in whom appropriate material was available, revealed the presence of small percentages of aneuploid cells in the marrow during the chronic phase of the disease and not seen with the routine procedure. These aneuploid cells may ultimately constitute the dominant clone during the blastic phase of the chronic myelocytic leukemia (CML). Furthermore, karyotypically abnormal cells, in addition those observed on routine study, were detected in the blastic phase when a large number of cells was examined. The value and implications of these observations are discussed.


The value of reverse banding in detecting bone marrow chromosomal abnormalities: translocation between chromosomes 1, 9, and 22 in a case of chronic myelogenous leukemia (CML). A case of chronic myelogenous leukemia (CML) with complex chromosomal abnormalities is reported. Conventional staining techniques indicated incorrectly that the Ph1 chromosome was not present. These studies showed a 46,XY,-1,+C karyotype in all bone marrow cells. Employing RFA (R bands by fluorescence using acridine orange) technique it was clear that the part of the long arm of chromosome 1 (1q23 leads to qter) was missing the tip of the long arm chromosome 22 (band q22) was translocated to it. The missing long arm of chromosome 1 was translocated to the long arm of chromosome 9. Furthermore, there was a break at band 9q22 and the whole terminal part was lost. The value of RFA technique is discussed.


Therapeutic considerations in acute lymphocytic leukemia. Evidence that the first human neoplasm systematically explored with chemotherapeutic treatments has apparently been cured in a palpable segment of affected patients evokes optimism for other types of cancer. The application of similar effort, similar logic, and quantitative experimental therapeutic approaches to the common cancers augurs well for cancer research and clinical medicine.


Gilles de la Tourette's syndrome, tardive dyskinesia, and psychosis in an adolescent.


Adriamycin--friend and foe.

Adriamycin--friend and foe.


The need for orthodontic treatment.

The need for orthodontic treatment. Basic problems of the need and demand for orthodontic treatment have been discussed and new areas of research have been identified. Discrepancies have been described between the views of the dental profession and the potential patients on malocclusion and the need for treatment. For malocclusions with or without impairment of the function the self perception of the patient may be different from and of more relevance than the professional judgment of the orthodontist. Before going into orthodontic treatment, the social sufficiency of the patient should be evaluated.


A radiographic study of tongue posture at rest and during the phonation of /s/ in class III malocclusion. In this investigation a group of individuals with Angle Class III malocclusions were studied radiographically to determine their tongue posture at rest and during phonation of /s/. Their speech and hearing was also evaluated. All of the subjects had normal hearing, while eleven of the twelve in the group showed some degree of sibilant distortion. At rest, these Class III subjects demonstrated a tongue posture that was lower than normal and more retruded. During the phonation of /s/ these individuals also demonstrated retrusion of the tongue in an apparent attempt to achieve a normal relationship of the tongue tip to the maxillary anterior teeth, an increased amount of mandibular depression, and a greater than normal distance between the tongue tip and the mandibular anterior teeth. Despite the compensatory movements exhibited by these individuals with Class III malocclusions, normal /s/ production was found in only one of the twelve subjects studied. This is in contrast to the successful compensations that result in normal /s/ production seen in a great number of individuals with a Class III, Division 1 malocclusion, as demonstrated by Subtelny, Mestre and Subtelny.


A new approach to cephalometric analysis of the dental base relationship. 1. The Reference line, connecting the centres of area of the projected skull outline, the cranium and the face, is relatively easy to locate and forms a more stable base from which to measure changes in the face in profile including the dental base relationship, than are the sella-nasion, maxillary or mandibular planes. 2. The Reference-Profile angle is a reliable assessor of a bimaxillary prognathism or retrognathism. Its mean value is 44 degrees. Angles above this value are associated with a bimaxillary retrognathism, and angles below this value with a bimaxillary prognathism. 3. The Reference-Dental Base angle is the most consistent for assessing the anterior jaw relationship between the maxilla and mandible. Its mean value of 50 degrees indicates a Class I relationship. Angles of 53 degrees and above indicate clinical Class II cases and angles of 48 degrees and below indicate clinical Class III cases. 4. These two angles are potentially valuable diagnostic indices for use in orthodontic clinical assessment.


Hyoid changes following orthopedic treatment of mandibular prognathism. In this study 30 patients exhibiting mandibular prognathism were treated by the orthopedic chin-cup appliance for a three-year period. The patients began treatment at the age of six years and completed treatment at the age of nine years. As previously reported, chin-cup treatment resulted in a "clockwise" facial rotation with the mandible moving relatively posteriorly and inferiorly. Hyoid position measured in this study also tended to move slightly posteriorly but was primarily displaced in an inferior direction. The positional alteration exceeded in amount what might have been expected by growth alone. The inferior direction of change in hyoid and associated structures and the resultant lack of encroachment on the vital pharyngeal passageways suggests that stability and patency of the pharyngeal airway is a primary factor in hyoid positioning. The duration and "growth guidance" aspects of treatment allowed for functional as well as morphologic adaption to the altered hyoid position.


Predicting angulational development and eruption of the lower third molar.


The effect of image quality on the identification of cephalometric landmarks. The effect of rare-earth phosphor intensifying screens on diagnostic image quality was examined. Five observers were asked to identify ten landmarks on each of fifteen cephalometric images having varying screen-film-kvp combinations. The most sensitive screens were found to significantly affect diagnostic image quality and thereby the reliability of landmarks identification. This finding was, however, only established for those landmarks having the lowest standard deviations and the clinical significance is thus questionable. There would appear to be other more important factors than physical image quality involved in the reliability of landmark identification. High intensifying screens are, therefore, recommended for use in cephalometric analyses since they help to reduce patient dose and do not appear to sacrifice diagnostic image quality to the extent that the reliability of cephalometric measurements is significantly affected.


Upper headgear versus lower headgear, yokes, and class III elastics. The use of a lower headgear-yoke-Class II mechanism on an average works faster than a maxillary headgear. The distal tipping of the molars is greater with HGY-II which may be the result of the quicker tooth movement. The speed of distalization is probably due to the effect of the constant force of the Class II elastics. An orthopedic change on the maxilla could be seen if one can draw any conclusions from a change of only 1 degrees. A reduction of the SNA angle and a slight clockwise rotation of the palatal plane was noticed. These results agree with earlier investigations about cephalometric changes with headgear therapy. We have come to the conclusion that both methods are indicated. We use the upper headgear especially at a time when a complete therapy is not possible because of noneruption of the permanent teeth. The HGY-II mechanism is indicated when most permanent teeth are available and thus the therapy can be completed in one treatment period.


Prediction of breadth of permanent canines and premolars in the mixed dentition. Multiple stepwise regression was used in the selection of mixed dentition variables capable of predicting the total breadth of the unerupted permanent canine and premolars. The material consisted of 77 children. Stone casts were made before and after eruption of the canine and premolars. At the first examination when the children were, on the average, 10 years old, intraoral roentgenograms were obtained of the canine and the premolars. To predict the total breadth of the upper canine and premolars the buccolingual breadth of the upper first permanent molar and measurements on roentgenograms of the breadths of the upper canine and premolars proved most useful. In the prediction of the total breadth of the lower canine and premolars the best results were obtained with measurement of the breadths of the teeth in the roentgenograms. The breadth of the incisors proved less useful as a predictor of the breadths of unerupted canines and premolars.


The mandibular dental arch: part II. Effects of lower incisor position on the soft tissue profile. It has been demonstrated that there are reliable methods of predicting the ultimate position of the lower incisor and the soft tissue. However, the concept of treating this tooth to any one number for every individual was not supported by the evidence. An optimum system would allow the clinician to select and combine factors from all methods and arrive at a balance of esthetics, function, and stability, obtaining a suitable compromise for his individual patient. This is undoubtedly attempted intuitively by many experienced clinicians.


Digital image enhancement of cephalograms. Quantification of cephalometric X-rays requires the use of hand tracings of the structures to delineate points, lines, and angles. The structures, especially soft tissue, are often obscure and are therefore approximated. Recent biomedical application of image enhancement by digital computer promises a method whereby landmarks in the cranium are more clearly defined. Development of this technique will enable a more extensive and accurate utilization of cephalometric radiographs for diagnostic and research purposes.


Is partial anodontia a syndrome of Black Americans? A study was designed to test the prevalence of congenitally missing permanent teeth in two Black American samples of children, ages 3-18. When combined, male with female, and compared with the Caucasian sample, the incidence of congenital absence is 2.57% and 5.15% for Caucasians.


Chronic, multiple tics of Gilles de la Tourette's disease. CSF acid monoamine metabolites after probenecid administration. Central nervous system metabolism in six children and one adult with the syndrome of chronic multiple tics was studied by measuring the accumulation of acid metabolites of dopamine and serotonin (homovanillic acid [HVA] and 5-hydroxyindole-acetic acid [5-HIAA], respectively) in the CSF following probenecid administration. The accumulation of 5-HIAA was reduced in patients with multiple tics in contrast with other pediatric patients (N = 27). The degree of reduction in 5-HIAA relative to HVA appeared to be associated with the severity of the tic disorder. With dextroamphetamine, tic symptoms worsened, CSF HVA level decreased, and CSF 5-HIAA concentration increased. These findings suggest an association in Gilles de la Tourette's disease of reduced functioning of inhibitory serotonergic mechanisms and functional dopaminergic overactivity.


A comparison of the rates of synthesis and turnover of collagen and non-collagen proteins in adult rat periodontal tissues and skin using a microassay.


Strain in mandibular alveolar bone during mastication in the rabbit.


The relationship between the surface area of the enamel crowns of human teeth and that of the dentine-enamel junction.


Electron microscopic study of the interaction of oral microorganisms with polymorphonuclear leukocytes.


In-vitro study of 65Zn uptake in developing rat enamel.


Ultrastructure and degeneration analysis of the nerve fibres of the tooth pulp in the cat.


Electron microscopy of epithelio-mesenchyme intercellular communication in trans-filter cultures of rat tooth germs.


An autoradiographic study of cellular proliferation of the mandibular condyle after induced dental malocclusion in the mature rat.


Ultrastructure of the innervation of the ducts within the rabbit parotid gland.


alpha-L-Fucosidase activity in human saliva and dental plaque.


Tongue function in the salamander Bolitoglossa occident alis.


Increase of extracellular cathepsin D activity in gingival washings during experimental gingivitis in man.


Contribution of different carbons of glucose to volatile acids during glucose catabolism in a human salivary sediment system.


Cortical structure of the pig mandible after the insertion of metallic implants into alveolar bone.


A quantitative radiochemical study of ionic and molecular transport in bovine dental enamel.


Diurnal periodicity of von Ebner growth lines in pig dentine.


Some electrochemical characteristics of human tooth enamel.


Effect of orally applied vaccines on oral colonization by Streptococcus mutans in rodents.


Negative interaction of orthophosphate with glycolytic metabolism by Streptococcus mutans as a possible mechanism for dental caries reduction.


Fluorescence histochemical demonstration of melanocytes in human buccal mucosa.


Sodium and potassium content in human palatine gland secretion.


Transformation of streptococci to streptomycin resistance by oral streptococcal DNA.


Blood flow in human mandibular elevators at rest and during controlled biting.


The less-acidic glycoproteins of the organic matrix of human dentine.


Fine structure of contacts between ameloblasts and odontoblasts in the rat tooth germ.


Endo- and exoglycosidases in an experimental rat osteosarcoma. Glycosidases capable of degrading intercellular matrix components were investigated in a 32P induced rat osteosarcoma. Homogenates of ossifying tumour were shown to readily degrade hyaluronic acid, chondroitin sulphates 4 and 6 but not dermatan sulphate. High levels of the exoglycosidases, beta-glucuronidase and beta-N-acetylglucosaminidase were found in tumour homogenates, and it was demonstrated that these enzymes contribute to the degradation of high molecular weight hyaluronic acid. The levels of these enzymes were compared with activities found in homogenates of neonatal bone and muscle surrounding tumours. Exoglycosidases, but not hyaluronidase, were found to be produced by cultures of osteosarcoma in vitro.


Analgesic abuse, renal parenchymal disease and carcinoma of the kidney or ureter. In a consecutive series of 88 cases of carcinoma of the kidney and upper urinary tract seen at one hospital, 31 had malignant urothelial tumours of the renal pelvis or ureter. Forty-two per cent of these transitional-cell carcinomas occurred in patients with renal papillary necrosis following upon prolonged and heavy analgesic ingestion. Other possible aetiological factors were heavy cigarette smoking (61% of cases), long standing urinary obstruction or infection (23%) and possible occupational exposure (6%); in only four cases (13%) was there no identifiable aetiological factor. Those cases with analgesic nephropathy were characterised by renal functional impairment, hypertension and interstitial nephritis, but there was no difference in the clinical behaviour or pathological appearances of the tumours in the two groups. The clinical and experimental evidence that certain metabolites of phenacetin are carcinogenic is reviewed.


Thiamine status in alcoholism.

Thiamine status in alcoholism.


Cigarette smoking in a cohort of Sydney schoolchildren: 1971--1974. Cigarette smoking habits of a cohort of Sydney schoolchildren aged 12--13 years in 1971 have been recorded for four years. Smoking prevalence increases steadily over those years, 2.3% of boys and 0.5% of girls smoking 20 or more cigarettes a week at age 12.7 years in 1971, rising to 23.6% of boys and 17.6% of girls by age 15.6 years in 1974. A substantial increase in smoking prevalence in children aged 15.6 years was found in 1974 compared with a similarly aged group studied by the National Health and Medical Research Council (Australia) Subcommittee on Smoking in 1968. Quite a few children changed their smoking habits during the study. Children who changed or left schools smoked more than children who remained at the one school. Attacks of asthma or bronchitis were not related to smoking except that girls who were very heavy smokers reported more of these illnesses in 1974. No changes in lung function could be attributed to smoking by age 14.6 years. New approaches to smoking need to be evaluated in view of its rising prevalence, especially among older schoolchildren.


"View from the bench".

"View from the bench".


The effect of ethyl alcohol on striated muscle: some clinical and pathological observations. Electrophysiological and pathological studies have been performed on three patients with recurrent focal swelling of the calf muscles simulating deep venous thrombosis, and in a patient with both cardiomyopathy and skeletal muscle disease. In all patients there were elevated CPK levels, histological evidence of an acute myopathy, heavy alcohol intake prior to the development of symptoms, and improvement in hospital with the cessation of alcohol consumption. These patients illustrate a form of alcoholic muscle disease which may be more common than generally realised.


Physiological responses to maximal exercise in a normal Australian population---comparative values in patients with anatomically defined coronary artery disease.