Effect of estrogen on gene expression: purification of vitellogenin messenger RNA. We report initial studies on estrogen-mediated regulation of egg yolk protein synthesis in the rooster. Egg yolk proteins are normally synthesized as a large precursor, vitellogenin, in the liver of the laying hen; roosters synthesize vitellogenin only when treated with estrogen. Polysomal RNA from the liver of estrogen-treated roosters was translated in a reticulocyte cell-free system, and the newly synthesized proteins were identified by a highly specific and sensitive indirect immunoprecipitation reaction. The messenger RNA that specifies vitellogenin has been purified more than 800-fold from rooster liver polysomal RNA by a combination of methods, including immunoprecipitation of polysomes and chromatography of RNA on poly(U)-Sepharose.


Bacterial luciferase requires one reduced flavin for light emission. Recent reports revive a hypothesis that the bacterial bioluminescence reaction involves two reduced flavin mononucleotide molecules per enzyme turnover. A two-flavin mechanism requires that the two flavins bind simultaneously or sequentially to the same or different sites on luciferase during a catalytic cycle. Measurements using equilibrium techniques show that the luciferase dimer has only a single reduced flavin binding site. Quantum yield results demonstrate that bioluminescence requires only one reduced flavin per luciferase, ruling out mechanisms involving either two reduced flavins or one reduced flavin plus one oxidized flavin per catalytic cycle.


Intramolecular crosslinking of tropomyosin via disulfide bond formation: evidence for chain register. Rabbit skeletal muscle tropomyosin can be crosslinked in the native state by the use of 5,5'-dithiobis(2-nitrobenzoate), which forms disulfide bonds between the two subunits. Using polyacrylamide gel electrophoresis in sodium dodecyl sulfate we have shown that this crosslinking goes to completion over a wide range of protein concentration, ionic strength, and reagent concentration. Crosslinks are not formed in denaturing solvents such as sodium dodecyl sulfate and guanidine hydrochloride despite the fact that the same number of SH groups react as in the native state (2.3 +/- 0.2). The sedimentation coefficients of crosslinked and non-crosslinked samples are identical. Thus, crosslinks are formed between corresponding cysteines on different chains of the same molecule. This provides strong evidence for a model of chain interaction with both chains in register. Evidence has also been obtained that rabbit skeletal tropomyosin is composed only of alphaalpha and alphabeta subunits rather than a random mixture of chains.


Nonribosomal proteins associated with eukaryotic native small ribosomal subunits. The native small ribosomal subunit (Sn) from rabbit reticulocytes which is able to initiate translation of globin mRNA in a cell-free system carries additional protein components. The latter can be separated from the subunit in a high salt sucrose gradient yielding a top fraction (T) and a complex fraction (C), sedimenting at about 4 and 15 S, respectively. Polyacrylamide gel electrophoresis in sodium dodecyl sulfate revealed that fraction T contained four dominant polypeptides, while fraction C represents a large protein complex consisting of at least 10 polypeptides. Sn isolated from other sources showed similar patterns of their nonribosomal proteins. Reconstitution experiments revealed that fraction C is absolutely required for protein synthesis, while fraction T enhances protein synthesis only in the presence of C. The adherence of these protein factors to the subunit is not mediated by magnesium ions. Treatment of Sn with EDTA and centrifugation in a magnesium-free sucrose gradient caused unfolding of the subunits and dissociation of several ribosomal proteins, but not of the factors. The unfolded ribosomal subunits sedimented as two distinct peaks. The more slowly sedimenting peak contained proteins of fraction T and the faster sedimenting one contained the 15S complex, indicating heterogeneity of the Sn population with respect to the factors attached to them.


Modification of intensity and direction of electron flow across bileaflet membranes. The intensity and the direction of electron flow across chloroplast extract bilayer membranes are modified by the presence of biliproteins. Biliproteins including C-phycocyanin, phycoerythrin, allophycocyanin, and Chroomonas phycocyanin enhance the photosensitivity of the membranes. The photoelectric spectra of the membranes in the presence of biliproteins correlate with the absorption spectra of these biliproteins. Experiments are presented to demonstrate the specific character of the electron-directing ability of biliproteins. A possible mechanism involving the interaction between the biliproteins and the membranes is proposed.


Fatty acids as modulators of membrane functions: catecholamine-activated adenylate cyclase of the turkey erythrocyte. Activation of the adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1[ from turkey erythrocytes by isoproterenol decreased precipitously below 26 degrees. Certain unsaturated fatty acids enhanced the activation by isoproterenol up to 25-fold at reduced temperatures. The fatty acid also enhanced the formation of a persistent active state of the enzyme which was produced by preincubation with guanosine 5'-(beta,gamma-imino)triphosphate [Gpp(NH)p]. Once the enzyme had been activated by Gpp(NH)p plus isoproterenol the reaction rate was no longer as temperature sensitive and the fatty acid had little effect. The synthetic Gpp(NH)p apparently substituted for the natural GTP, which is known to play a regulatory role in the adenylate cyclase system. The findings suggest that the function of GTP which is mediated by the hormone is the temperature-sensitive event which is enhanced by the fatty acid. The use of free fatty acid to probe membrane-associated reactions in intact cells and in isolated membrane preparations is proposed.


Mechanism of activation of adenylate cyclase by cholera toxin. Cholera toxin (choleragen) can stimulate adenylate cyclase [EC 4.6.1.1; ATP pyrophosphate-lyase (cyclizing)] activity in whole particulate fractions or purified plasma membranes of homogenates of isolated fat cells provided special precautions are taken to stabilize the enzyme during the required preincubation period. As observed with intact cells, the activation exhibits a protracted (about 25 min) lag phase, and it is blocked by ganglioside GM1 and choleragenoid ("binding" subunit of toxin). The 36,000 molecular weight subunit ("active" subunit), a hydrophobic polypeptide which does not block choleragen binding or action, can directly activate the enzyme in intact cells without a lag phase. Its effects are not blocked by ganglioside GM1 or choleragenoid, yet the stimulated activity exhibits reduced fluoride and enhanced isoproterenol sensitivity, properties characteristic of the choleragen-activated enzyme. Binding of the 125I-labeled 36,000 molecular weight subunit to cells is not saturable and is unaffected by gangliosides, choleragen, or choleragenoid, and the bound material behaves as an integral membrane protein; this protein may simply partition into the membrane matrix. With increasing time of incubation cell-bound choleragen may dissociate into its component subunits, but these remain in the membrane. Using a double antibody immunoprecipitin system, substantial precipitation of cyclase activity occurs with antisera against the 36,000 molecular weight subunit provided toxin activation has occurred. The normal process of activation may involve an initially inactive toxin--ganglioside complex which, as a result of lateral mobility and multivalent binding (lag phase), results in destabilization of the molecule with release of the "active" subunit into the membrane core where it can spontaneously associate with and perturb the cyclase complex.


Regulation of binding properties of the nicotinic receptor protein by cholinergic ligands in membrane fragments from Torpedo marmorata. Exposure of receptor-rich membrane fragments from Torpedo marmorata to carbamylcholine causes a slow (half-time of 5--10 min) and reversible change of properties of the cholinergic receptor protein manifested by a decrease of the initial rate of Naja nigricollis alpha-[3H]toxin binding in the presence of carbamylcholine. This change corresponds to a 5- to 20-fold increase of affinity for carbamylcholine. Other agonists, acetylcholine, phenyltrimethylammonium, show the same effect but not the antagonists d-tubocurarine and flaxedil. Decamethonium and hexamethonium show little, if any, agonistic effect in vitro on the same membrane fragments but cause the affinity change. This regulatory property can be lost after aging of the preparation of membrane fragments. Since the affinity increase progresses with a similar time course as the decrease of amplitude of the permeability response consecutive to agonist preincubation, it is proposed that, in the membrane at rest, the receptor protein is present under a state of low affinity for agonists and that the reversible stabilization by the agonists of a high affinity state corresponds to the "pharmacological desensitization" of the system as predicted by one of the models of Katz and Thesleff.


Substrate specificity of the cyclic AMP-dependent protein kinase. The protein substrate specificity of the catalytic subunit of rabbit skeletal muscle cyclic AMP-dependent protein kinase (EC 2.7.1.37; ATP:protein phosphotransferase) has been studied using genetic variants of beta casein. It was found that beta casein-B was phosphorylated at a much greater rate than beta caseins A1, A2, A3, or C. The enhanced phosphorylation of beta casein-B, as compared with the most common variant A2, was attributed to an arginine substitution for a serine at position 122, which caused a nearby residue, serine 124, to become a phosphorylation site for the protein kinase. These results further support the concept that the local primary structure is important in specificity and that arginine may be a specific determinant common to all the local phosphorylation site sequences recognized by the cyclic AMP-dependent protein kinase.


Degradation of penicillin G to phenylacetylglycine by D-alanine carboxypeptidase from Bacillus stearothermophilus. D-Alanine carboxypeptidase from Bacillus stearothermophilus is a membrane-bound enzyme which is inhibited by covalent interaction with penicillin G. The penicilloyl enzyme spontaneously reactivates and simultaneously releases a penicillin G degradation product; 0.2 mumol of the latter was isolated after incubation of 4.2 mumol of [8-14C]penicillin G with 10 g of membrane protein. It was identified as phenylacetylglycine by chromatographic techniques, infrared spectroscopy, and mass spectrometry. A mechanism for the degradation is proposed in which the remaining part of penicillin G would be released as 5,5-dimethyl-delta2-thiazoline-4-carboxylic acid. The implications of this finding are discussed.


A spike-forming model of the neural membrane. A model of the neural membrane is described, based on a set of "convertible gates" on its inner surface. The atomic arrangement of a gate is specified to be subject to modification by a set of chemical reactions, sometimes permitting sodium ions, sometimes potassium ions, sometimes neither, to have access to the transmembrane channel it guards. Reaction rate and current/voltage equations are developed, values assigned to the parameters, and computer solutions obtained. Under pulse or continuous stimulating current the model is found to generate action potential spikes closely similar in height and shape to those of real neurons. Other electrical properties are also relatively neuron-like.


Headgroup conformation and lipid--cholesterol association in phosphatidylcholine vesicles: a 31P(1H) nuclear Overhauser effect study. The nuclear Overhauser effect has been observed in the nuclear magnetic resonance spectra of 31P. The information content of the nuclear Overhauser effect has been applied to the structure and dynamic properties of phosphatidylcholine vesicles. In the vesicles only 1/3 of the theoretical maximum nuclear Overhauser effect enhancement is observed. This result is accounted for by dipolar interactions between the N-methyl protons and the phosphate of phosphatidylcholine, and a correlation time for internal motion of 1.4 X 10(-9) sec. Addition of up to 30% cholesterol does not change the nuclear Overhauser effect enhancement or spin-lattice relaxation time of the vesicles. It is argued that the OH group of cholesterol is hydrogen bonded to the ester carbonyl oxygen of the phosphatidylcholine molecules.


Nuclear coupling of 33S and the nature of free radicals in irradiated crystals of cysteine hydrochloride and N-acetyl methionine. The 33S hyperfine structure has been observed in the electron spin resonance of irradiated crystals of cysteine-HCl-H2O and of N-acetyl DL-methionine. An analysis of the results shows that in both substances the free radicals that are stable at room temperature are disulfide radicals, RCH2SS. A possible mechanism for formation of these radicals is proposed.


Relaxation measurements on the acetylcholine receptor. In Electrophorus electroplaques, the agonist-induced postsynaptic conductance depends on membrane potential. During steady exposure to agonists, after a voltage step the conductance relaxes on a millisecond time scale, exponentially approaching a new equilibrium value. The relaxation rate constant k is an instantaneous function of voltage, insensitive to the past or present conductance. Two components sum to form k. A concentration-sensitive component increases linearly with agonist concentration and decreases during desensitization or exposure to curare. Thus this component reflects the average frequency at which acetylcholine receptors are opening. The voltage-sensitive component, obtained by extrapolating k to zero agonist concentration, increases at more positive potentials. For acetylcholine, the voltage-sensitive component equals the rate constant for the exponential decay of postsynaptic currents; it thus seems to be the closing rate for active receptors. The voltage-sensitive component has the relative amplitudes acetylcholine less than carbamoylcholine less than decamethonium, and for each agonist equals the closing rate determined from "noise" measurements at neuromuscular junctions. The kinetic data explain several aspects of the steady-state conductance induced by agonists, but shed no light on apparent cooperative effects.


Quasi-elastic behavior of solutions of viral capsid and RNA at very low shearing stresses. By the application of shearing stresses on the order of 10(-3) dyne cm-2 (10(-2) muN cm-2), via the magnetic viscodensimeter, extremely high relative viscosities (greater than 500) were observed when turnip yellow mosaic virus was degraded in alkali into its capsid and RNA. The solutions, however, possessed a watery consistency at this stage and exhibited a quasi-elastic character by rotor-recoil experiments. The development of this curious behavior was concentration and temperature dependent; it was not seen less than 0.5% nor at 8 degrees, and appeared sooner at 30 degrees than at 20 degrees. The time of appearance was delayed as the pH was lowered; however, the effect was still observed when the pH was as low as 9. Whereas reversibility was demonstrated when the shearing stresses exceeded the elastic resistance [0.17 dyne cm-2 (1.7 muN CM-2)], thorough mixing usually resulted in a normal behavior of the solutions thereafter. Values for the modulus of rigidity at 20 degrees for about 1% virus concentration was less than 2 X 10(-2) dyne cm-2 rad-1 (0.2 muN cm-2 rad-1), which, while extremely small, was reproducible. A porous structure, possibly involving a capsid and RNA complex, is envisioned.


The glass point of elastin.

The glass point of elastin. Elastin undergoes a glass transition in a temperature range depends on its water content. This behavior is similar to that of amorphous polymers swollen with solvent and, therefore, is additional evidence for the random network model proposed for the structure of elastin.


Myelin structure transformed by dimethylsulfoxide. X-ray diffraction patterns from nerves bathed for about one-half hour in Ringer's solution containing dimethylsulfoxide at concentrations of 10% or more show reflections from a new, highly ordered structure with a repeat period about two-thirds that of native myelin. The proportion of myelin transformed is greater at higher concentrations, and above 40% the native pattern is no longer observed. Replacing the dimethylsulfoxide with Ringer's solution leads to the rapid reappearance of the native diffraction pattern. The effect of dimethylsulfoxide can be accounted for by the loss of water from the spaces between the membrane units without significant modification of the bilayer structure.


Mechanism of removal of senescent cells by human macrophages in situ. Mechanisms by which macrophages discriminate between "mature-self" and "senescent-self" were investigated using the human red blood cell (RBC) system as a model. Conditions simulating those encountered in situ were adhered to as closely as possible by using short term culture techniques and incubating with autologous macrophages and immunoglobulins (Ig). It was found that RBC aged in vitro were phagocytized when they were incubated in pooled, normal human IgG, allogeneic Ig, autologous IgG or Ig, washed with medium, and then incubated with autologous macrophages. RBC treated in the same way but incubated in IgM or Iga, Ig-depleter serum, or Medium 199 alone were not phagocytized. This indicates that Ig is required for phagocytosis and suggests that the Ig which attaches to RBC is IgG. When freshly drawn RBC were separated into young (Y) and old (O) RBC according to density and incubated with autologous macrophages, less than 5% of the Y-RBC were phagocytized, whereas greater than 30% of the O-RBC were phagocytized, independent of whether the final incubations were performed in medium without serum (Y-RBC, 5 +/- 2% phagocytized; O-RBC, 33 +/- 1.5%), or autologous Ig-depleted serum (Y-RBC, 2 +/- 2.5%; O-RBC, 51 +/- 17%), or whole serum (Y-RBC, 0%; O-RBC, 43 +/- 5%). This indicates that (1) the Ig is attached in situ to the old RBC, and (2) that phagocytic recognition is not inhibited by other serum components. Scanning electron microscopy, employing labeled anti-IgG, IgM, and IgA reagents, revealed that Y-RBC had essentially no Ig on their surface, whereas old RBC had IgG on their surface. These findings indicate that IgG attaches in situ to senescent human RBC, making them vulnerable to phagocytosis by macrophages.


Cell surface species-specific high affinity receptors for discoidin: developmental regulation in Dictyostelium discoideum. Vegetative (noncohesive) D. discoideum cells and cells at several stages during the development of cohesiveness were fixed with glutaraldehyde, and their agglutinability by purified carbohydrate-binding proteins (lectins) from slime molds  and plants was determined. The two purified lectins from D. discoideum, called discoidin I and II, were poor agglutinins of fixed vegetative D. discoideum cells and potent agglutinins of fixed cohesive D. discoideum cells. The increased agglutinability of D. discoideum cells by discoidin I or discoidin II with differentiation paralleled discoidin appearance in cell extracts and development of cohesiveness. The lectin of another slime mold, Polysphondylium pallidum (pallidin) and the lectin of Ricinus communis (RCA-I) were also more potent agglutinins of fixed differentiated D. discoideum cells than of fixed vegetative D. discoideum cells. However, wheat germ agglutinin was much more potent with fixed vegetative cells. The association constant (Ka) of discoidin I and discoidin II for fixed differentiated D. discoideum cells was in the range of 10(9) M-1, more than twenty-fold higher than the Ka of these lectins for fixed vegetative cells. The Ka of discoidin I and discoidin II for fixed differentiated P. pallidum cells was an order of magnitude lower than for fixed differentiated D. discoideum cells. The Ka of pallidin for fixed differentiated P. pallidum cells was about 4 X 10(9) M-1 and was an order of magnitude lower for fixed differentiated D. discoideum cells. Since previous studies showed that the lectin of each of these slime mold species is detectable on the surface of cohesive cells, the observation of species-specific receptors on the surface of cohesive cells suggests a basis for species-specific cohesion.


Erythroid colony induction without erythropoietin by Friend leukemia virus in vitro. Erythroid colonies could be produced without the addition of erythropeietin in plasma cultures seeded with bone marrow cells from normal C3Hf/Bi mice by exposure of the cells in vitro to medium from a cell line (IS) that continuously produces Friend leukemia virus in culture. The activity in the culture medium was viral rather than erythropoietin-like, since it was sedimentable by high-speed centrifugation and heat labile. Erythroid colonies did not develop when the bone marrow cells exposed to virus-containing medium were from mice genetically resistant to Friend virus. IS culture medium contained both Friend spleen focus-forming and XC-plaque-forming activities. No erythroid colonies were induced when genetically sensitive cells were exposed to a preparation from which the spleen focus-forming activity had been removed, but which contained XC plaque-forming activity in high concentration. Thus the spleen focus-forming component of Friend virus appeared to be responsible for inducing erythroid colony formation without erythropoietin in vitro. Some erythroid colonies were also found in control cultures to which neither virus nor erythropoietin had been added. Reduction in the concentration of fetal calf serum in the culture medium substantially decreased the number of these colonies but had only a minor effect on the number of virus-induced colonies. The number of erythroid colonies produced after 2 days of culture without erythropoietin or fetal calf serum was approximately proportional to the titer of Friend spleen focus-forming virus to whcih the bone marrow cells had been exposed. This system should prove useful for investigation in vitro of Friend virus--host cell interactions which lead to erythropoietin-independent erythropoiesis.


Stimulation of epinephrine-sensitive fat cell adenylate cyclase by cytosol: effect of cholera toxin. Cytosol prepared from rat epididymal fat cells by centrifugation at 100,000 X g for 1 hr was found to enhance the basal and epinephrine-sensitive adenylate cyclase [EC 4.6.1.1; ATP pyrophosphate-lyase (cyclizing)] of fat cell ghosts. Cholera toxin also stimulated adenylate cyclase and increased the response to epinephrine in fat cells. A possible relationship between the adenylate cyclase modifying activities of cytosol and the effects of cholera toxin was sought. Cytosol from freshly prepared fat cells added to ghosts prepared from cells that had been exposed to toxin for varying periods showed a progressive loss of responsiveness to cytosol epinephrine-enhancing activity. The effect appeared within 15 min after toxin exposure, a full 30 min before any direct effect of toxin on adenylate cyclase was seen. Since exposure to toxin decreased membrane response to cytosol epinephrine-enhancing activity, the possibility that epinephrine-enhancing activity in cytosol might be altered by toxin was explored. Cytosol from cells exposed to toxin for varying periods lost epinephrine-enhancing activity to an appreciable degree within 15 min. Examination of these early events after exposure to toxin should clarify the way in which this bacterial substance affects mammalian cells. The cytosol epinephrine-enhancing activity was destroyed by boiling for 3 min and was partially inactivated by trypsin. It was nondialyzable and stable at -70 degrees.


Inhibition of tubulin assembly by RNA and other polyanions: evidence for a required protein. Nonneural cell extracts contain a heat-stable, nondialyzable activity that will inhibit the spontaneous assembly in vitro of partially purified brain tubulin. The sensitivity of this inhibitory activity to ribonucleases but not to a variety of other hydrolytic enzymes indicates that the inhibitor is an RNA. This conclusion is supported by the observation that purified RNAs from sea urchins, chinese hamster ovary cells, and brain all inhibit spontaneous microtubule assembly in vitro. The synthetic polynucleotides [poly(A), (C), (G), and (U)] are also inhibitory. This inhibition, however, appears to be nonspecific since the RNA base composition is unimportant and a variety of other nonnucleic acid polyanions also function as inhibitors. The treatment of assembly competent tubulin preparations with an insoluble RNA in the form of poly(A) covalently linked to agarose beads produces a "stripped" tubulin which will not assemble microtubules unless a heat-stable, trypsin-sensitive fraction eluted with increased ionic strength is mixed with the "stripped" tubulin. Similar results can be obtained with other cation exchangers, including phosphocellulose and carboxymethylcellulose. The heat-stable protein sequestered by poly(A)-agarose appears to be identical to the "tau" factor recently described by Kirschner and coworkers. Reconstitution experiments indicate that there is a stoichiometric requirement for these factors. These results suggest that spontaneous assembly of microtubules in nonneural cell extracts is blocked because the endogenous factors are complexed with RNA. This idea is supported by the observation that the ratio of tubulin to RNA is low in cultured cell extracts but very high in neural tissue extracts.


Brain tryptophan hydroxylase: purification of, production of antibodies to, and cellular and ultrastructural localization in serotonergic neurons of rat midbrain. Tryptophan hydroxylase [EC 1.14.16.4; L-tryptophan, tetrahydropteridine:oxygen oxidoreductase (5-hydroxylating)], the enzyme catalyzing the rate-limiting step in the biosynthesis of serotonin, was purified 79-fold from the region of the raphe nucleus of rat midbrain by sequential column chromatography and disc-gel electrophoresis. In electrophoresis three bands were distinguished, A, B, and C, which, when separated and submitted individually to electrophoresis, reproduced the same three bands. Bands A and C were enzymatically active and inhibited by para-chlorohenylalanine. Antibodies produced to each of the three bands crossreacted by immuno double diffusion and electrophoresis with each other and homogenates of raphe nuclei; they completely inhibited enzyme activity only of tryptophan hydroxylase. Tryptophan hydroxylase was localized by light and electron immunohistochemistry to serotonin neutrons of the raphe. Ultrastructurally, in cell bodies, the enzyme was distributed in cytoplasm and in association with endoplasmic reticulum and Golgi apparatus. In dendrites and axons, it was associated with microtubules. Tryptophan hydroxylase in brain is only neuronal and cytoplasmic, exists in multiple forms, and is associated with microtubules, suggesting it may be transported from sites of synthesis in cell body into axons.


Relationship between nuclear and cytoplasmic poly(adenylic acid). The kinetics of accumulation of radioactive poly(A) in the nucleus and cytoplasm of mouse L cells have been determined using labeling conditions in which the cellular ATP pool is shown to have a nearly constant specific radioactivity. Most or all nuclear poly(A) accumulates with kinetics very similar to those of heterogeneous nuclear RNA, having a half-time of about 25 min. There is little or no lag before attainment of a maximal rate of accumulation of cytoplasmic poly(A). These data are consistent with a variety of models in which nuclear poly(A) does or does not all serve as a precursor of cytoplasmic poly(A). In either type of model there must be a class of poly(A) either synthesized in the cytoplasm or passing through a small nuclear pool separate from the main pool of nuclear poly(A).


Normal genetically mosaic mice produced from malignant teratocarcinoma cells. Malignant mouse teratocarcinoma (or embryonal carcinoma) cells with a normal modal chromosome number were taken from the "cores" of embryoid bodies grown only in vivo as an ascites tumor for 8 years, and were injected into blastocysts bearing many genetic markers, in order to test the developmental capacities, genetic constitution, and reversibility of malignancy of the core cells. Ninety-three live normal pre- and postnatal animals were obtained. Of 14 thus far analyzed, three were cellular genetic mosaics with substantial contributions of tumor-derived cells in many developmentally unrelated tissues, including some never seen in the solid tumors that form in transplant hosts. The tissues functioned normally and synthesized their specific products (e.g., immunoglobulins, adult hemoglobin, liver proteins) coded for by strain-type alleles at known loci. In addition, a tumor-contributed color gene, steel, not previously known to be present in the carcinoma cells, was detected from the coat phenotype. Cells derived from the carcinoma, which is of X/Y sex chromosome constitution, also contributed to the germ line and formed reproductively functional sperms, some of which transmitted the steel gene to the progeny. Thus, after almost 200 transplant generations as a highly malignant tumor, embryoid body core cells appear to be developmentally totipotent and able to express, in an orderly sequence in differentiation of somatic and germ-line tissues, many genes hitherto silent in the tumor of origin. This experimental system of "cycling" teratocarcinoma core cells through mice, in conjunction with experimental mutagenesis of those cells, may therefore provide a new and useful tool for biochemical, developmental, and genetic analyses of mammalian differentiation. The results also furnish an unequivocal example in animals of a non-mutational basis for transformation to malignancy and of reversal to normalcy. The origin of this tumor from a disorganized embryo suggests that malignancies of some other, more specialized, stem cells might arise comparably through tissue disorganization, leading to developmental aberrations of gene expression rather than changes in gene structure.


Receptor synapses without synaptic ribbons in the cochlea of the cat. This report describes recepto-neural junctions thought to be chemical synapses without synaptic ribbons or vesicle aggregates. Such synapses occur in the cat between the outer hair cells of the cochlea and auditory nerve fibers. These synapses are formed by flat or indented junctions having symmetric membrane complexes associated with smooth endoplasmic reticulum and coated vesicles in the outer hair cell cytoplasm. Inner hair cells have receptoneural junctions with asymmetric membrane complexes; synaptic ribbons and vesicles gather at one type of junction, while the other type associates with endoplasmic reticulum. Gap junctions are not seen at cochlear synapses but do link supporting cells. The results suggest that inner and outer hair cell synapses have different properties and raise questions about the role of synaptic vesicles and endoplasmic reticulum in the storage and release of transmitters.


Hemoglobin Cranston, an unstable variant having an elongated beta chain due to nonhomologous crossover between two normal beta chain genes. An asymptomatic woman was found to have a compensated hemolytic state due to an unstable hemoglobin variant, comprising 35% of the total. Peptide maps of tryptic digests of the abnormal beta chain were identical to those of beta A except that tryptic peptide 15 (Tyr-His-COOH) was absent and a new peptide was detected, containing equivalent amounts of Ser, Ile, Thr, and Lys. Its sequence was determined by manual Edman degradation. An additional hydrophobic peptide isolated by counter-current distribution contained: Asx, Ser, Ala, Tyr, 2 Phe, and 3 Leu. Its sequence was determined on an automatic solid phase sequencer. Digestion with carboxypeptidase A confirmed the C-terminal sequence. Hemoglobin Cranston has an elongated beta chain with the following structure: (see article). This variant is the first "adult" human hemoglobin known to contain isoleucine. It is likely that hemoglobin Cranston arose because of a nonhomologous crossover of two normal beta chain genes, probably during meiosis, with the insertion of two nucleotides (AG) at position 144, resulting in a frame shift. Hemoglobin Cranston provides new information on the structure of the beta chain gene as well as an explanation of both the structure and genetic basis of hemoglobin Tak, the only other elongated beta chain variant that has been described.


Nucleotide sequences of the 3'-terminal untranslated region of messenger RNA for human beta globin chain. In normal messenger RNA for the human beta-globin chain, nucleotide sequences have been identified which can be matched to the amino-acid sequence of the abnormally long segment of the beta-chain of hemoglobin Cranston. The finding of these sequences strengthens the hypothesis that the betaCranston chain arose by a frameshift mutation allowing the "readthrough" of the normal termination codon and translation of usually untranslated portions of the messenger RNA for the beta-globin chain. The oligonucleotides which match the amino-acid sequence of hemoglobin Cranston provide a sequence of 36 nucleotides which follows the normal beta-chain termination codon UAA.


A mechanism for initiation of genetic recombination. A mechanism for the initiation of genetic recombination is proposed. Its key features are the pairing, nicking, and cross-annealing of palindromic loops, i.e., structures formed by DNA with sequences of inverted complementary repeats. This mechanism may provide a simple, yet specific means of producing crossed strand connections between homologous DNA duplexes to form structures which can be intermediates in the process of genetic recombination.


Transposition of R factor genes to bacteriophage lambda. Transpositions of segments of R factor (antibiotic resistance plasmids) to bacteriophage lambda have been selected and characterized. Cells of Escherichia coli harboring R factors that determine kanamycin resistance were infected with phage lambda, and lambdakan transducing lines were obtained. Each of the three examined is unusual when compared to lambda transducing phages containing E. coli chromosomal genes: the kan insertions (a) occur at several sites, each well removed from the integration region POP', (b) are not associated with deletion of lambda phage DNA, and (c) are separable from the lambda genome during transduction or during lytic growth. Two insertions from the same R factor contain 1.5 kilobase sequences repeated in inverted order. The properties of the lambdakan phage suggest that R factors contain systems capable of mediating genetic exchange in the absence of extensive DNA homology. It is suggested that such systems of exchange may have played important roles in R factor evolution.


Expression of the beta-thalassemia gene in the first trimester fetus. To determine whether beta-thalassemia can be detected in the fetus, blood was obtained from abortuses of normal mothers and of mothers with beta-thalassemia trait. The red cells were incubated with radioactive leucine and the globin chains were analyzed by radiochromatography. Two independent methods were utilized to correct the results for contamination by maternal radioactive beta-chain, and the corrected beta/gamma ratios were compared to a previously established range of normal fetal beta/gamma synthetic ratios obtained by similar measurements in pure fetal cells. In the erythroid cells of three fetuses from mothers with beta-thalassemia trait, the beta/gamma synthetic ratio was normal in two. The third had a beta/gamma ratio of 0.04 at 10 1/2 weeks, a 50% reduction, consistent with fetal beta-thalassemia trait. Two other fetuses, derived from parents both of whom had beta-thalassemia trait, were also studied. One had a beta/gamma ratio of 0.029 at 8 weeks, a 65% reduction, also consistent with beta-thalassemia trait. The cells of the other had a ratio of essentially zero at 11 weeks, highly suggestive of homozygous beta-thalassemia. Although further experience will be needed to distinguish the homozygous and heterozygous states reliably, it now appears that the beta-thalassemia gene is expressed in the first trimester. Therefore these data suggest that the antenatal diagnosis of beta-thalassemia is becoming an attainable goal.


Analysis of incipient reproductive isolation within a species of Drosophila. Drosophila willistoni and its subspecies quechua are morphologically indistinguishable, but differ in relative frequencies of certain allozyme genes. The cross quechua female X willistoni male produces nearly or completely sterile males, while the reciprocal cross gives fertile males. Hybrid females are fertile. Analysis of the sterility with the aid of chromosomes marked with mutant genes shows that the second and X-chromosomes have major and third chromosomes minor effects. Backcross males of the same chromosomal constitution may be either fertile or sterile; the threshold effect may be due to environmental variations or to gene variants present in the strains crossed. Only a trace of ethological isolation between the subspecies is present. In contrast to a narrowly localized and geographically isolated subspecies bogotana of D. pseudoobscura, the subspecies quechua of D. willistoni is fairly widespread, although its geographic area is not accurately known. Neither with bogotana nor with quechua are there valid reasons to suppose that these subspecies are of very recent origin.


Mutants of T7 bacteriophage inhibited by lambda prophage. Mutants in gene 20, a new T7 gene, cannot grow on rex+ lambda lysogens. Gene 20-- mutants suppress in double mutants the phenotype of T7 ligase negative mutations, but not vice versa. Amber 20- mutants have been obtained. There are differences between these T7 mutations and the similar T4 rII mutations. There are host mutations which permit T7 20- mutants to grow on lambda+ lysogens. T7 DNA synthesis on normal lambda+ lysogens infected with 20- mutants is essentially normal, but the DNA is not packaged. The gene 20 protein is active in in vitro complementation and probably used late in infection for DNA packaging into phage heads.


Selection for conditional gametogenesis in Chlamydomonas reinhardi. A new technique is described for selection of temperature-sensitive mutants affecting gametogenesis in Chlamydomonas reinhardi. The first mutant found by this technique is characterized. Cells exhibiting the gam-1 phenotype are capable of sexual agglutination, but cannot form zygotes at the restrictive temperature. The mutation, however, has been expressed only in gametes of mating type (-). Cells of mating type (+) which carry this gene are able to engage in normal zygote formation. Temperature shift experiments and antibiotic studies have indicated that this gene is expressed within 6 hr after the onset of gametogenesis in liquid culture, and its product has a lifetime of about 4 hr at 35 degrees.


Complementation of H-2-linked Ir genes in the mouse. The immune response to the random linear terpolymer of L-glutamic acid, L-lysine, and L-phenylalanine (GLphi) is under dominant H-2-linked Ir gene control in the mouse. Matings between two nonresponder strains produced responder F1 hybrids, demonstrating complementation of the nonresponder alleles. This observation, coupled with the fact that several intra H-2 recombinant strains derived by recombination between two nonresponder parental haplotypes are also GLphi responders, indicated at least two dominant loci are concerned with responsiveness to this terpolymer. The complementary genes were termed alpha (tentatively localized in a new subregion of the H-2 complex, I-F) and beta, which maps in the I-A subregion. Generally, both the alpha(+) and beta(+) alleles are required for responsiveness. However, in the (C57BL/6J X SJL)F1 hybrid we noted complementation between two parental nonresponder strains, each of which carried beta genes derived from different H-2 haplotypes, yet lacked functional alpha genes. The possible cell levels at which these genes may function in the regulation of the immune response are discussed.


Synthesis of hemoglobin AIc in normal and diabetic mice: potential model of basement membrane thickening. Adult diabetic mice (C57Bl/KsJ--db/db) have increased amounts of a minor hemoglobin in their peripheral blood compared to wild-type (+/+) mice. This increase is analogous to the 2-fold increase of a glycohemoglobin with similar chromatographic mobility (Hb AIc) seen in the blood of patients with diabetes mellitus. Although the exact chemical nature of human or mouse Hb AIc is unknown, both contain a sodium-borohydride-reducible linkage on the beta chain which is a presumed Schiff base between a sugar moiety and the protein. The db/db animals, which have normal amounts of mouse Hb AIc at weaning, show the increase approximately 4 weeks after the onset of the signs of diabetes. This rise is brought about by an increase in a circulating factor that determines directly or indirectly the synthesis of mouse Hb AIc as a post-synthetic modification of Hb A. Evidence for this was obtained by showing that the rate of synthesis of the modified Hb is linear for at least the first 50 days of the life of the red cell and that the rate of synthesis is dependent on the environment in which the cells circulate. Thus the rate of mouse Hb AIc synthesis in +/+ cells is greater when those cells circulate in a db/db host than when they circulate in a +/+ host. The nature of the humoral factor is unknown. If glycosylations of basement membrane proteins and hemoglobin proceed via a common mechanism, then the monitoring of Hb AIc could provide a useful model for studying the early events of basement membrane thickening.


[13C]Valine metabolism in methylmalonicacidemia using nuclear magnetic resonance: propinonate as an obligate intermediate. [Alpha-13C]- and [alpha,beta-13C]valine were administered sequentially to a patient with methylmalonicacidemia to clarify the metabolic pathway of valine from methylmalonic acid semialdehyde to methylmalonyl-CoA. Methylmalonic acid was isolated from multiple urine samples, purified, and analyzed by 13C nuclear magnetic resonance spectroscopy. Contrary to the widely accepted view, the results show unequivocally that methylmalonic acid semialdehyde is decarboxylated to propionate before conversion to methylmalonyl CoA.


Mating signal and DNA penetration deficiency in conjugation between male Escherichia coli and minicells. Strain OU122, a dnaB mutant of HfrH which stops vegetative DNA synthesis immediately when the temperature is shifted to 43 degrees, was mated at 43 degrees with minicells with and without 1 mM Zn2+. Synthesis of DNA was detected in the mating mixture containing minicells derived from either Fchi925 or F+chi115 cells, but only a small amount was detected in the mixture containing 1 mM Zn2+ which inhibits the formation of mates (previously called mating pairs). The supernatant liquid from cell cultures did not induce DNA synthesis, suggesting that DNA synthesis was not stimulated by diffusible molecules. Additional experiments showed that Zn2+ inhibited DNA synthesis associated with DNA transfer but did not inhibit the DNA transfer that had already been initiated. Thus, the stimulation of DNA synthesis observed required physical interaction of cells and F pili. Reisolated minicell exconjugants derived from the cross OU122 X F--chi925 minicells were shown to contain transferred DNA in contrast to either the cross OU122 X F--chi925 minicells in 1 mM Zn2+, or the cross OU122 X F+chi1115 minicells. Thus, F+chi1115 minicells retained the property of surface exclusion at 43 degrees.


Rapid changes in rat pineal beta-adrenergic receptor: alterations in l-(3H)alprenolol binding and adenylate cyclase. The properties of the beta-adrenergic receptor which regulates adenylate cyclase [ATP pyrophosphate-lyase (cyclizing)8 EC 4.6.1.1] in the pineal gland are similar to the properties of the sites which specifically bind l-[3H]alprenolol, a potent beta-adrenergic antagonist. Stimulation of the beta-adrenergic receptor results in a 30-fold increase in the activity of N-acetyltransferase (= arylamine acetyltransferase; acetyl CoA:arylamine N-acetyltransferase, EC 2.3.1.5), an enzyme involved in the synthesis of thepineal hormone melatonin. In the normal diurnal light-dark cycle there is greater physiological stimulation of the beta-adrenergic receptor in the pineal during the night than during the day. Pineals from rats kept in constant light for 24 hr possess more hormone-sensitive adenylate cyclase and specifically bind more l-[3H]alprenolol than do pineals from rats kept in the dark overnight. When rats, exposed to light for 24 hr, are treated with the beat-adrenergic agonist isoproterenol, there is a rapid loss of both hormone-sensitive adenylate cyclase activity and specific l-[3H]alprenolol binding sites. There is no change in the affinity of adenylate cyclase for isoproterenol or for its substrate, ATP. Similarly, although there are fewer binding sites, there is no change in the affinity of the remaining sites for either agonist or antagonist. Inhibition of protein synthesis with cycloheximide does not affect the loss of either adenylate cyclase activity or specific binding sites. The data suggest that stimulation of the beta-adrenergic receptor causes a rapid decrease in the number of available receptors and in hormone-sensitive adenylate cyclase activity; conversely, lack of stimulation causes an increase in these parameters. It is suggested that these changes contribute to the phenomena of super- and subsensitivity in the pineal gland by regulating the capacity of the pineal to synthesize cyclic AMP in response to beta-adrenergic stimulation.


Control of meal size by central noradrenergic action. Previous investigations of central noradrenergic effects on food intake have concentrated on the use of high doses of noradrenaline (two to 200 times brain noradrenaline content). In this work we examined the effect of low doses of noradrenaline (not exceeding brain noradrenaline content) on the parameters of spontaneous ingestive behavior. By arranging for rats to trigger remote infusions of noradrenaline into their own anterior forebrains at the beginning of spontaneously initiated meals, meal size was very reliably increased more than 200% by doses of 0.015--0.37 nmol (2.5--62 ng of noradrenaline base) (n = 12). The effect was alpha-adrenergic. It was blocked by phentolamine but not by propranolol. Infusions of noradrenaline at the above doses, which nearly triple meal size, did not elicit eating when made during an intermeal interval, nor did they influence the length of the intermeal interval when made 60 min after the termination of an uninfused meal. These results show that noradrenaline increased food intake at doses less than 1% of the brain's endogenous noradrenaline content. Meal size is more susceptible to alteration by noradrenaline manipulations than is meal frequency.  The brain's own noradrenergic system may function to sustain food intake once feeding is initiated. This function of brain noradrenaline may control spontaneous meal size.


An effect of castration and testosterone replacement on a circadian pacemaker in mice (Mus musculus). Castration of mice in freerunning conditions (total darkness, DD) causes a reduction of running wheel activity in the beginning of the active period (alpha) and stimulates activity at the end of alpha. Simultaneously, the period (tau) of the freerunning rhythm is increased. Both effects are abolished by implantation of a Silastic capsule from which a physiological dose of testosterone is released at a constant rate. The results are tentatively explained by differential endocrine influences on two oscillating components in the pacemaker of the circadian activity rhythm.


Group selection, altruism, reinforcement, and throwing in human evolution. Evolution of altruism by group selection involves sacrifice of some individuals, not to the "group as a whole," but to other individuals in the group. Deme-group selection may establish strictly altruistic genes in a population, but only under limited conditions, and perhaps never among vertebrates, among which apparently altruistic behaviors may always potentially benefit the altruists. Responsive-group selection is a more effective mode of evolution of altruism, conspicuous in man. Evolutionary reinforcement increases the force of selection of advantageous behaviors, including altruistic ones, by making them pleasant or rewarding. It is probably involved also in ecological habitat selection, and may be the source of many human emotions, including esthetic ones. Throwing (of stones and weapons) exemplifies both the possible importance of a difficult-to-measure evolutionary factor and the role of reinforcement; in human evolution throwing may have been decisive in food-getting and fighting, in shifting emphasis from brute force to skill, and in inducing evolution of a brain able to handle three-body geometric problems precisely and thus preadapted for more complex functions.


Cytoenzymochemical effects of some antiblastic drugs and prediction of response to chemotherapy in acute leukemias. Over 20 cytoenzymochemical tests were carried out in 152 patients with different types of acute leukemia to estimate the effects of some antiblastic drugs such as L-asparaginase, Purinethol, Methotrexate, Endoxan, Vinchristine, Cytosine Arabinoside a.o. The patients selected for the study were carefully examined before treatment at different moments during and/or at the end of the treatment. The effects of these drugs on the blast cells were mild when the cellular populations had a low rate of nucleic acid synthesis, high glycogenic score and high amounts of lipids or an important oxidative enzymatic activity. The enzymatic prediction tests: the acid phosphate deviation test and the succinic dehydrogenase inhibition test including the variant suggested by some of the authors - the latic dehydrogenase inhibition test - gave satisfactory results only in certain cases of acute leukemia.


[Chemistry and pharmacology of glucocorticosteroids]. The glucocorticosteroids are derived chemically from androsterone or pregnane. Their natural models are cortisone and cortisol (or hydrocortisone), characterised by the presence of oxygen or an oxydryle group at 11, of a --CO -- CH2OH group in the 17 position and of an-OH group in the 17 position. Their anti-inflammatory power dominate their pharmacodynamics: it goes hand in hand with their power to increase gluconeogenesis. They have an immunodepressant effect whose power seems to be proportional the their anti-inflammatory action. The use of hydrocortisone in the treatment of shock remains empirical. The mechanism of their anti-allergic effect remains debated. Glucocorticosteroids disturb lipid and protein metabolism, water and electrolyte balance, the central nervous system, and gastric secretion; they inhibit the secretion of ACTH. Their pharmaceutical forms are numerous: they are adapted to the oral route and to the parenteral routes (i.m., i.v., i.r., intra-articular) and to external use (cutaneous and mucosal applications).


[Secondary side effects of corticosteroids and the management of corticosteroid therapy]. The side effects of glucocorticoids are the result of their biological effects and closely related to the dose administered and the duration of treatment. Whilst excessive amounts may result in the development of a clinical and biological Cushings syndrome, there exists also the problem of avoiding acute adrenal insufficiency when corticosteroids are stopped after a prolonged period of administration or during substitutive therapy for adrenal insufficiency. Aspects requiring surveillance are discussed.


[Complications following subchondral, not intra-articular, injections of corticosteroids]. The author describes two cases of bone changes following non-intra-articular and more probably sub-chondral injection of corticosteroids. The first, in condyle a mandibular was reversible with reconstruction of the bone after a year. The second, in the condyle of a knee was definitive with a permanent chip in the bone.


[Action of corticosteroids on cutaneous cicatrization. Use in maxillofacial surgery]. The effects of glucocorticoids on cutaneous cicatrisation are discussed. It is generally admitted that they have a delaying effect especially when applied locally. Their use in maxillo-facial surgery is limited: generally, they are used against oedema; locally, to check excessive fleshy growth and to treat cheloid scars.


[Results of a survey on the usefulness of adding corticoids to root canal obturation pastes]. 500 questionnaires were sent out. The statistical survey was carried out on 116 replies. It emerges from this survey that 81% of our colleagues who answered use a canal filling paste containing a corticoid. The reason for its use is the prevention of fairly serious medicamentous arthritis. A small number (5 colleagues) had complications which they ascribed to the associated corticoid. A few use a non-hardening paste containing a corticoid as a temporary canal filling.


[Corticotherapy and stomatologic oncology]. Long-term use of corticoids had proved beneficial in the treatment of advanced stomatological cancers, the effective dose being 40 mg of prednisone per day. In so far as it is possible to distinguish any specific action on individual symptoms, its action on pain was obvious in 40% of cases, on inflammation and infection in 52% of cases on fever (independent of infection) in 14% of cases, on oedema in 34% of cases. It was almost invariably favourable on the general condition. The method of using it emphasize how easy it is to use corticoids by injection which is of particular value in our speciality in view of: dysphagia, administration of large doses, deficiencies in the general condition, sometimes a matter of urgency. The counter-indications usually recognized for corticoids need to be modified in the case of cancer patients. Incidents occur surprisingly rarely and are minor relative to the advantages of the therapy, provided the patient is kept under strict supervision.


[Corticotherapy and mucocutaneous pathology]. The tremendous advances in treatment brought about by corticotherapy applied to cutaneo-mucosal pathology should not be allowed to obscure the fact that its action is merely palliative, that it should only be proceeded with after careful diagnosis and that it may trigger undesirable side-effects. General corticotherapy is definitely indicated in certain serious dermatoses (e.g. pemphigus vulgaris) in large doses at the beginning of the course of treatment which often has to be kept up indefinitely; it is in these patients that the most serious accidents occur. It is also indicated in other dermatoses (e.g. lichen planus) in smaller doses and in separate courses, generally triggering incidents and accidents of a less serious nature which to a certain extent seem to be attenuated by taking the drug on alternate days. It is counter-indicated in one particular condition: psoriasis. Corticotherapy by intra- and sub-lesional local injection is most useful in the treatment of certain localised skin lesions (e.g. cheloids) and of the oral mucosa (e.g. erosive lichen planus). Either a few drops are injected or a larger quantity in a suspension of microcrystals. Complications have sometimes been observed in the skin (leukoderma, dermoepidermatrophia and, particularly, amaurosis), but never so far after sub-mucosal injections. Local corticotherapy by external application, very widely used in the form of ointments, creams and lotions for numerous cutaneous conditions may cause various more or less serious local side-effects, the systemic effects with depression of the hypophyso-adrenal axis, only seem to occur to any extent with occlusive dressings. It can also be used in the treatment of some conditions of the oral mucosa (e.g. some forms of lichen planus, benign mucous membrane pemphigoid) by means of either a corticosteroid incorporated into a special excipient which adheres to the mucous membrane or in tablets of 17-betamethasone valerate which gradually break up in the saliva. With the usual posology of 10 tablets of 0.1 mg per day, even over several months, there are no systemic effects, 17-betamethasone valerate (unlike phosphate) having an action which is primarily topic and being practically unabsorbed as has been shown by assessment of plasmatic cortisol.


Thymidine kinase activity in human bone marrow cells. The thymidine kinase activity per 10(6) DNA-synthesising marrow cells and the rate of incorporation of tritiated thymidine into the DNA of 10(3) DNA-synthesising marrow cells were estimated in 9 haematologically normal patients and 49 patients suffering from a variety of haematological disorders. Slight increases in thymidine kinase activity were found in 6 of the 31 patients with haematological diseases associated with normoblastic erythropoiesis and greater increases were found in 3 of the 18 patients with megaloblastic haemopoiesis due to vitamin B12 or folate deficiency. In the latter group, there was a statistically significant inverse correlation between haemoglobin levels and thymidine kinase activity. No correlation was found between thymidine kinase activity and the rate of incorporation of tritiated thymidine in either the normoblastic or megaloblastic group, suggesting that the level of thymidine kinase activity does not limit the rate of incorporation of exogenously supplied thymidine into the DNA of human bone marrow cells.


[Etiology and treatment of glossodynias]. This study only concerns classical glossodynia with few symptoms and not related to general, tumoral or local syndromes. Lingual pain is caused in the majority of cases by apraxic movements of the tongue. These movements, related to the anguish, can by a regressive process be related to the "searching" lingual movements of the child in psychological distress. The patient suffering from glossodynia (usually a woman) is enclosed in a viscious circle : anguish, apraxic movements, lingual pain, of which the beginning is often contemporary with a cancer in the area. Therapy should be addressed first of all at the distress, by looking very closely at the patient, by examining her very carefully, by absolutely gaining her confidence and by asserting the absence of cancer while admitting the reality of her disease. It should then be addressed to the lingual movements, by immobilization exercises giving rise to consciousness of movements up to then unconscious. Relaxation exercises can be combined. Failure of this therapy : frequent in men, in those who speak a lot (teachers), in hysterics "pithiatics", senile, and those persons whose intellectual evolution is slight. The author emphasizes the therapist's availability ; success in this treatment being related to what he can give of himself to these patients.


[Phonetic results of velopharyngoplasties. Apropos of 67 cases]. The phonetic results of velopharyngoplasty depend on the anatomical conditions of velopharyngeal reconstitution. But this is not the only condition and the results are variable depending on the degree of phonetic disorders and their longstandingness. The latter therefore account for a certain number of bad results whatever the surgical technique used.


[Root canal screws in amalgam restoration of devitalized teeth]. The placing of amalgam screws into root canals is indicated (1) on damaged teeth if the coronal retenion is insufficient, (2) on vital teeth, after canal treatment, as an alternative to dentine anchors like small crews or pins, and (3) for building up crown cores. Four systems of canal screws are described, Dentatus, FKG, Anthogyr and Mooser. The large differences of dimensions and conicity necessitate a better knowledge of the necessary dimensions of the root reamers and drills. After comparative evaluation of the systems, the clinical technique for the user of the Mooser system is demonstrated.


[Gingivitis in Zurich preschool children]. The gingival condition of 96 boys and 92 girls 5 years old was clinically examined using the sulcus bleeding index (Mühlemann)Son, 1971). 58% of all children exhibited a light degree of gingivitis. The average inflammatory degree was 0.17 (SBI). There was no significant difference in degree of inflammation between the boys and girls. Only 24 gingival units per child were inflammed.


[Motivation and education].

[Motivation and education]. It was thought for many years that successful motivation in oral health should be based on the knowledge of epidemiology of caries and periodontal diseases and the methods of prevention. One should learn from mistakes: 95 per cent of the Swiss population "suffers" from caries and periodontal disease but apparently this had little impact on prevention. Simple, world-wide accepted methods of prevention are not used by the majority although they provide a pronounced reduction of caries and gingivitis. The number of Swiss communities which offer adequate preventive programs to schoolchildren ranges between 5 to 10 per cent. Although the budgets of school dental services could be lowered by 75%. Reasons for such failures may be inadequate information of dentists, authorities and the population. Not enough information is spread regarding sugarless sweets, or prevention by fluoride. Unsuccessful motivation probably also lies in the inadequate assignment of the roles ofthe patient, his dentist and the social security. So far only the dentist has formulated oral health duties to the patient, e.g. a better oral hygiene, a reasonable healthy nutrition. In the future the patient should have the right to ask the dentist some questions. Is his dentition at the end of a treatment, healthy and ready to render hygienic efforts effective? Patient, doctor and insurances should know that disease still exists if gingival papillae bleed upon using a toothpick. Absence or presence of gingival bleeding are the criteria for health or disease. A check list of the state of oral health and a signed questionnaire concerning the patient's knowledge of oral health are used to improve motivation of both patient and dentist.


Differentiation of erythroleukemic cells in the presence of inhibitors of DNA synthesis. Erythroid differentiation can be readily induced by butyric acid in cultured erythroleukemic cells in the presence of inhibitors of DNA synthesis and in the absence of cell division. This result appears to rule out more complex models for globin gene expression which require gene replication or cell division (or both).


Incontinentia pigmenti (Bloch-Sulzberger syndrome) associated with acute granulocytic leukemia. Incontinentia pigmenti, a syndrome of developmental defects, was found to be associated with acute granulocytic leukemia in a 4-month-old black girl. She has been responsive to antileukemic drugs, and remission has been induced without difficulty after each relapse. Although survival time was expected to be very short because of her age, race, type of leukemia, and initial peripheral leukocyte count at time of diagnosis, she is still alive 35 months after diagnosis of leukemia. One may speculate that the prolonged survival time might be related to the unknown genetic defect.


[Immunological study in carcinogenesis. Distribution of tissue incompatibility antigens (system HL-A) in oncological patients]. By means of a micromethod of lymphocytotoxic test the content of eleven HL-A antigens (1, 2, 3, 5, 7, 8, 9, 10, 11, 12 and 13) was studied in 200 healthy human subjects, 100 patients with rheumatoid polyarthritis and 82 patients with bone tumors. The latter included 50 patients with benign tumors (osteoblastoclastomas, chondroblastomas, chondromas, non-osteogenic fibromas) and 32 patients with malignant tumors (chondro- and osteosarcomas, Ewing sarcoma, malignant osteoblastoclastomas). It was found that in patients with different bone tumors antigen HL-7 was encountered reliably more frequently than in control groups, in patients with malignant tumor also antigen HL-A10 was more often detected. The most frequently observed haplotyes in oncological patients were HL-A3/7 and HL-A2/7. In patients with rheumatoid polyarthritis antigen HL-A3 was found more rarely and antigen HL-A10 more frequently than in healthy subjects. The most frequent haplotypes in this group were as follows: HL-A2/7, HL-A2/8 and HL-A11/12. The possible mechanisms of the relationship between HL-A antigens and the development of pathology are discussed. Some considerations are offered concerning the possibility to utilize HL-A typing for an accessory differential diagnosis.


[Left ventricular decompression by means of a paracorporeal transatrial ventriculo-aortic bypass pump: hemodynamics and long-term results (author's transl)]. Circulatory assist devices in current clinical use are analyzed. In this study the hemodynamic effects and long-term results with a paracorporeal ventriculo-aortic membrane pump with transatrial access to the left ventricular chamber are reported. The effectiveness of the pump was demonstrated by left ventricular decompression (-63%) and reduction of MVO2 (-41%), TTI (-71%), dp/dt max (-66%) and Q cor (-17%). These highly significant hemodynamic changes were, furthermore, confirmed by evaluation of the force-velocity relation (V max). In chronic experiments the bypass pump was implanted in 6 calves. The longest survival time was 18 days. 5 experiments were terminated on account of rupture of mechanical parts, one death was due to acute renal failure caused by emboli. Different bypes of ventriculo-aortic assist devices are discussed in regard to technical problems, hemodynamic efficiency and long-term results.


Breast disease. A clinical study with special reference to diagnostic procedures. A consecutive series of 1.310 women with breast complaints and a control group of 241 healthy women were examined by clinical examination and mammography. The majority of the palpable and non-palpable lesions were preoperatively classified on a four-point scale according to the degree of suspicion of malignancy. A similar classification was performed after mammography as well as after aspiration biopsy. Some aspects of the case-histories including a positive family history of breast carcinoma, marital status and breast feeding were studied in women with breast complaints and compared to the findings in a control group of women. Previous breast disease, mode of detection, delay and breast symptoms were also studied...


Electron microscopic observations of the utricle and ampullae in a case of dizziness of suspected saccular origin. Electron microscopic observation was reported on the utricle and ampullae of a case, a 36-year-old male, with dizziness attack due to suspected saccular origin. After the removal of the utricle and ampullae, the dizziness recurred, and finally the saccular nerve section relieved the dizziness attack of this case. So the actual lesion was localized in the saccule and was not in the specimens obtained at the first surgery. Though most of hair cells appeared normal, there were a few with many vacuoles or shrinkage of the cytoplasm distributing sparsely throughout the sensory epithelia. The distribution of the pathological hair cells in the utricle was more general than in the ampullae. The sensory hair cells without pathology and the presynaptic structures were similar to those found in the experimental animals. Nerve and nerve endings showed normal structures. The findings may be physiological at this age group or an extension of the pathology of the dizziness-causing lesion.


[Lesions of the vertical and horizontal systems in the optokinetic vestibular system]. Symptoms of vestibulo-ocular disturbances may be classified into two major groups; i.e., vertical plane and horizontal plane disturbances. Vertical plane symptoms arise from midline lesions of the cerebellum or the brainstem and also from diffuse or symmetrical lesions to either. Vertical plane symptoms also arise from lesions which simultaneously effect both peripheral vestibular systems. Horizontal plane symptoms originate from a unilateral disturbance in the vestibular system. Thus, vertical plane symptoms indicate bilateral disturbances and the symptoms appear symmetrically in the eyes, trunk or extremities. Vertical plane symptoms appear in the sagittal plane, or the plane of the vertical semicircular canals. On the other hand, horizontal plane symptoms appear in the horizontal plane, or the plane of the horizontal semicircular canals.


Transition of nystagmus types in unilateral labyrinthine diseases. In order to clarify changes in spontaneous and positional nystagmus which occurred during the course of peripheral labyrinthine diseases, 31 patients with Ménières disease (17 cases), aural vertigo of unknown etiology (9 cases), and sudden deafness (5 cases) were tested repeatedly using ENG. The testing of each case was made 4.8 times on the average with an average interval of 7.2 days. Twenty-four cases (77.4%) showed more than two of the following four types which are listed in serial order of occurrence: spontaneous (or direction-fixed positional) nystagmus toward the involved ear; geotropic direction-changing positional nystagmus; spontaneous (or direction-fixed positional) nystagmus toward the opposite ear; and apogeotropic direction-changing positional nystagmus. Thus, the above-mentioned order may be regarded as the uniform pattern in the nystagmus transition caused by acute unilateral labyrinthine diseases.


Tubing, as a treatment for recurrent infant suppurative otitis media. (1) The details described below have evidenced that the tubing therapy is an influential treatment for the recurrent infant suppurative otitis media. The tube with a beveled head and split shank is very convenient and effective for the treatment since it is not only easily inserted but also easily removed, preventing it from migration into the tympanum. (2) The treatment must promote the drainage of discharges through the infant auditory tubes which are thick and short and improve the ventilation of the middle ear cavity half permanently. (3) According to the author's experience, the spontaneous extrusion of the ideally set tube after 2 or 3 months is a sign of a perfect cure of this disease.


Responses of cortical vestibular center produced by stimulation of vestibular nuclei and visual association area in the cat. (1) The cortical projection of the vestibular nuclei (LVN, MVN and IVN) was investigated by the evoked potential and unit discharge analyses in the cat anesthetized with alpha-chloralose and/or immobilized with gallamine triethiodide. (2) This projection field coincided well with that of the vestibular nerves, being principally contralateral by LVN stimulation but nearly symmetrical by either MVN or IVN stimulation. (3) Of 11 units responsive to vestibular nuclei stimulation, 6 reacted to stimulation of the visual association cortex (anterior part of the Clare-Bishop area). The response pattern of the cortical vestibular units by C-B stimulation was a sequence of excitation, inhibition and rebound. (4) Interaction of visual information with the cortical vestibular neurons was discussed.


A quantitative test of optokinetic nystagmus and its data processing by computer. A quantitative test of optokinetic nystagmus was proposed. Horizontal nystagmus was examined by a large rotating cylinder with vertical stripes. Nystagmus responding to the cylinder rotation with an acceleration of 2 degrees/sec2 for 90 seconds was recorded with an electronystagmograph. On the records, the number of beats, average eye-speed, and average amplitude per each 10 seconds were calculated. This method was suitable for testing the nature of optokinetic nystagmus which indicates the ability of the eyes to adapt to objects moving through the visual field. Moreover, in order to save trouble in hand-scoring of each parameter of nystagmus, computer processing of electronystagmographic data was introduced. The results were printed numerically on the teletypewriter, and displayed graphically on a cathode ray tube and X-Y recorder. By displaying measured values on the form already printed with the normal ranges, the evaluation of the results was performed readily and objectively.


Optic organ and cervical proprioceptors in maintenance of body equilibrium. Two experiments were made on the functional correlation between the optic organ and the cervical proprioceptors in maintenance of body equilibrium. Experiment I: The effects of weak and strong electric stimulation of the deep nuchal muscles on changes in optokinetic nystagmus and optokinetic after-nystagmus were examined in adult non-albino rabbits with binocular vision. Experiment II: Experiment I was repeated on rabbits with monocular vision giving optokinetic stimulation moving from the side of the blindfolded eye to that of the open eye and applying only weak electric stimulation to the neck muscles. The results obtained were as follows: (1) Experiment I: Weak electric stimulation of the deep nuchal muscles of rabbits with binocular vision tended to promote optokinetic nystagmus with either no significant change or decrease in optokinetic after-nystagmus. Strong electric stimulation had the opposite effects, i.e., it tended to inhibit optokinetic nystagmus and activate optokinetic after-nystagmus. It also produced abnormal optokinetic nystagmus, such as the inversion phenomenon of optokinetic nystagmus and "optokinetic nystagmus firing". (2) Experiment II: Weak electric stimulation of the deep nuchal muscles of rabbits with monocular vision had similar effects to those in experiment I, but promotion of optokinetic nystagmus was less obvious, while activation of optokinetic after-nystagmus was more marked. In addition, the inversion phenomenon of optokinetic nystagmus developed even on weak electric stimulation. From these results the following conclusions were drawn: (1) The cervical proprioceptors can function in two mechanisms causing promotion and break down of equilibrium of the optokinetic system. (2) The cervical proprioceptors, as an organ of equilibrium, can exert two different actions on the optokinetic eye reflexes, i.e., they can promote optokinetic nystagmus and inhibit optokinetic after-nystagmus.


Neurotological studies on the role of the sympathetic nervous system in the formation of traumatic vertigo of cervical origin. To test the validity of the hypothesis that irritation of the cervical sympathetic nerves is a cause of hypertonicity of the cervical soft supporting tissues, especially the deep nuchal muscles, and that this hypertonicity is a cause of traumatic vertigo of cervical origin, we examined 44 vertigo cases with whiplash injury by testing equilibrium function and EMG discharges from the neck before and after administration of drugs affecting the sympathetic receptors. The results obtained were as follows: 1. Administration of iso-proterenol (beta-receptor stimulant) caused a significant increase in the EMG's from the injured neck muscles, in parallel with increased impairment of the righting reflex. The blindfolded vertical writing test showed the appearance or intensification of ataxia or deviation in writing when this drug was given. In contrast, administration of propranolol (beta-receptor depressor) had the opposite effects on the EMG's, the righting reflex and blindfolded vertical writing. Furthermore, the resulting changes in the EMG's and equilibrium functions were parallel with alterations in subjects' complaints, i.e., increase or decrease in neck pain and vertigo. 2. Administration of drugs affecting the alpha receptors, such as noradrenaline (alpha-receptor stimulant) and phentolamine (alpha-receptor depressor) had no appreciable effect on the EMG's, equilibrium function or the subjects' complaints. These results show that in vertigo due to whiplash injury hypertonicity of the cervical erector muscles can be induced sympathetically, and that this hypertonicity is based on over-excitement of beta receptors in the injured neck muscles, which results in vertigo of cervical origin.


Lumbomuscular proprioceptive reflexes in body equilibrium. To evaluate the role of reflexes related to the lumbar proprioceptors in maintenance of body equilibrium, changes in equilibrium function of the eyes and body were observed after unilateral procainization of the lumbar erector muscles. Observations were made on normal subjects and vertigo cases with lumbar pain after whiplash injury using various equilibrium tests. The results obtained were as follows: (1) On unilateral procainization of the lumbar erector muscles of normal subjects, eye nystagmus and disturbances of the righting reflex developed. Simultaneously, changes in drift reactions of the lower limbs were detected by the stepping test. Namely, in many of the subjects examined the direction of stepping deviation became quite different from that before procainization, and stepping after procainization tended to show slight or moderate ataxic features, associated with a sensation of unsteadiness. (2) When procaine was injected unilaterally into tender spots in the lumbar erector muscles of traumatic vertigo cases, spontaneous eye nystagmus and disturbances of the righting reflex decreased. Simultaneously, significant changes in the drift reactions of the lower limbs were observed in many of the cases examined. Namely, the direction of deviation became the opposite of that before procainization and ataxia in walking almost disappeared with reduction in vertigo. The following conclusions were drawn from these findings: (1) The effects of procaine on equilibrium of normal subjects are in sharp contrast to its effects on equilibrium of traumatic vertigo cases. Findings in the former might be due to increased imbalance between the activities of the right and left lumbar proprioceptors, while those in the latter might be due to decreased imbalance between the two. (2) These findings support the view that from the standpoint of body equilibrium, there are two phases of the proprioceptive reflex, and that Fukuda's concept of "two phases of the labyrinthine reflex, i.e., a stage of disturbance and a stage of coordination", can be applied to interpretation of the proprioceptive reflex of lumbar origin.


[Reaction of the middle ear mucous membrane in tubal obstruction]. 1. Tubal obstruction causes a rapid rise in transepithelial cytopempsis. The squamous epithelial cells show numerous vesicles carrying subepithelial fluid (extracellular fluid) to the middle ear cavity. 24 hours following tubal obstruction a widespread rupture of the epithelial junctions can be observed. Subepithelial fluid, consisting of lymphfluid, transsudate and later exsudate, enters the middle ear cavity without passing a cellular barrier. 2. Long lasting negative pressure provoces a cellular differentiation (metaplasia!) of the formerly squamous epithelium to a high, cylindric cell layer with new functions: ciliary movement and secretory activity. 3. Ablation of the tympanic membrane to expose the middle ear to air after 2-4 days, causes a stop in the epithelial differentiation but no regeneration to the normal histological flat formation. 4. Studies of samples of human middle ear mucosa in serous otitis or sero-mucous otitis show exactly the same results as during the different stages of our animal experiments.


Simultaneous application of electronystagmography (ENG) and photoelectronystagmography (PENG) in the case of thermal labyrinth examination. At examination of normal persons, the ENG registration method was compared with that of PENG. Special examinations were executed with regard to the behaviour of the beat rates during the culmination phase of thermal labyrinth reactions by using each of both registering methods. By synchronous recording of PENG and ENG, it was possible to settle the discrepancy of the greater number of beats in ENG compared with those of PENG, while the examinations were executed at different times. The PENG-method does not exclude an optical fixation due to the necessary interior illumination, and this results in a retarding of the reaction and smaller beat rates. This fact should be considered when comparing the results of ENG and PENG.


The effect of gravity on galvanic nystagmus in rabbits. The effect of gravity on a galvanic nystagmus provoked in rabbits was investigated. Weightlessness increased the amplitude but decreased the frequency as well as the speed of the nystagmus. During continuous galvanic stimulation the provoked nystagmus disappeared spontaneously after a period of 60-120 seconds. Weightlessness was able to arouse this nystagmus again. An explanation of this phenomenon might be that galvanic nystagmus undergoes a modifying effect from the vestibular maculae and their state of stimulation.


Tissue culture of the organ of Corti and the isolated hair cells from the newborn guinea pig. The organ of Corti and isolated hair cells from newborn guinea pig were cultured with the Rose chamber method. The hair cells could be maintained for more than 20 days after explantation when the organ of Corti was cultured as a whole, though the isolated hair cells swelled and degenerated within about 13 hours after explantation. By comparison between the tissue culture of the hair cells with the organ of Corti and isolated hair cell culture, the following conclusions were reached: (1) There are some functional correlations concerning the nutritional requirements and cell metabolism between the hair cells and the supporting cells in addition to the structural correlations. (2) There are no distinct fundamental differences between both inner and outer hair cells about their affectability as a single cell level.


Postural behavior and motion sickness.

Postural behavior and motion sickness. (1) An interesting style of acting was demonstrated on a stage of a Kyogen, a classic comedy of Japan, titled "Funawatashimuko", i.e., "A boatman and a bridgegroom in a boat". Antagonistic postures which move toward the opposite direction were displayed by a boatman who is pulling an oar and a passenger who is being moved by the rolling of a boat. (2) Why doesn't one suffer from motion sickness when he drives a car but may suffer from it when he is a passenger? This question was answered, from the standpoint of human postures, by observing the antagonistic postures exhibited by a bus-driver and a passenger, and also by the findings in postrotatory eye nystagmus (an indication of artificial motion sickness) which was varied according to the three different positions of the head. (3) Learning postural adjustment against motion sickness, developing through repetitively traveling in vehicles, was also objectively shown in a posture of an experienced lady bus-conductor whose body inclined in the same direction as that of the driver. Its similarity to the establishment of a kinetic labyrinthine reflex in chickens was explained.


Serum lysozyme activity in children with hematological and malignant disorders. Pretreatment serum lysozyme activity was high in 2 children with myelomonocytic leukemia, 800 and 1 000 mug/ml, normal in all 10 cases of acute lyelocytic leukemia and subnormal in 21 of 34 cases of acute lymphocytic leukemia. Normal values were found in all but one case of acute lymphocytic leukemia during complete remission including 8 cases after all therapy had been discontinued. All 8 are still in complete remission. Low serum lysozyme activity was found in 5 patients with acute lymphocytic leukemia in complete relapse, this could possibly be helpful in the diagnosis of early relapse. Activity was subnormal in 5 of 17 children with malignant tumours, and in 3 of 65 cases of various benign hematological disorders.


Liposarcoma a clinicopathologic, radiographic and prognostic study. The present investigation is a clinicopathologic, radiologic and prognostic study of 122 liposarcomas registered in the Swedish Cancer Registry between 1958 and 1966. The incidence of liposarcoma in Sweden was estimated to be 2.5 cases per million inhabitants per year. The liposarcomas were divided into 4 main subtypes according to WHO's recommended classification (1969): well-differentiated (33 cases), myxoid (22 cases), rundcell (14 cases) and pleomorphic types (21 cases). When predominantly well-differentiated or myxoid liposarcomas contained areas of round-cell or pleomorphic types (24 and 8 cases, respectively), they were classified separately, thus adding 2 further subtypes to our classification. The validity of this histologic classification was emphasized by the prognostic study, based on a 10 year follow-up. The differential diagnosis for each of the 4 main subtypes is discussed. Three well-differentiated, lipoma-like liposarcomas which contained highly vascular hibernoma-like areas are described. Two well-differentiated liposarcomas situated in the neck and upper part of the back, respectively, of elderly patients included area similar to "spindle-cell lipoma" (Enzinger). An intracytoplasmic network of birefringent fibers with the staining reaction of collagen was found in 2 well-differentiated fibrosing liposarcomas...


Meningeal leukemia in the blastic phase of chronic granulocytic leukemia. One hundred one patients were treated for Ph' positive chronic granulocytic leukemia (CGL) in the blastic phase. In seven of these (6.9 per cent), meningeal leukemia developed. Of the 99 patients who died of their disease, a complete remission was achieved in 12 with a median survival of 12 months (three to 28 months). Incomplete responders had a median survival of only 2.5 months (one to 14 months). In five of the 12 complete responders (42 per cent), but in only two of the incomplete responders (2.3 per cent), meningeal leukemia developed. The principal neurologic signs were cranial nerve palsies and papilledema. All patients had pleocytosis with myeloblasts in the cerebrospinal fluid. As in patients with acute leukemia and diffuse histiocytic lymphoma, increased survival of patients in whom hematologic remission from the blastic phase of CGL is achieved may allow sufficient time for the development of meningeal leukemia. Intrathecal methotrexate is extremely successful in treating this complication. Cerebrospinal fluid pleocytosis was eradicated in all seven of our patients, and neurologic symptoms and signs were completely eliminated in five patients. No evidence of meningeal leukemia was found in three of the five patients in whom an autopsy was performed.


The class II malocclusion: differential diagnosis and clinical application of activators, extraoral traction, and fixed appliances. The importance of an exact differential diagnosis of the Class II malocclusion and its indications for treatment with activators, extraoral forces, and fixed appliances has been shown. Four schematized malocclusions with their cephalometric characteristics and their individualized orthopedic therapeutics have been presented. Special attention has been given to the subsequent fixed mechanotherapy showing the limited need for fixed appliances. This approach is attractive, for it reduces considerably the stress imposed on the patient and the orthodontist, it bypasses the iatrogenic hazards entailed in extensive and prolonged mechanotherapy, and it consistently increases chances for long-term stability.


Remodeling of teeth by grinding.

Remodeling of teeth by grinding. Pulp and dentin reactions to extensive grinding of human teeth were studied clinically and histologically. The facial portions of forty-eight premolars to be extracted for orthodontic reasons were reshaped to "lateral incisors" using diamond instruments and abundant water cooling. Following grinding and polishing with sandpaper discs and pumice, topical fluoride was administered. The patients were asked to report any increased sensitivity reactions. The teeth were extracted at 0, 1, or 3 weeks or 3 to 5 months and examined histologically. No significant discomfort was reported by the patients except for an initial period of a few days during which there was increased sensitivity to temperature changes. The histologic findings indicated that extensive remodeling of young permanent teeth is possible with no or only minor pulp and dentin reactions. Some initial localized effects in terms of aspiration of odontoblast nuclei, hyperemia, absence of a cell-free zone, and slight cellular infiltration were sometimes observed, but the long-term observations indicated that these reactions were transient. Secondary dentin formation was not registered. It is important that gentle grinding techniques with abundant water spray be used and that smooth and self-cleansing surfaces which allow hypermineralization of the ground enamel and dentin be prepared. In a few teeth in which proximal retention areas were unintentionally prepared, caries developed and marked pulp reactions occurred. Therefore, great care must be taken not to introduce proximal steps when mesiodistal tooth width adjustments are performed. The present study has provided histological and clinical evidence to indicate that even extensive recontouring by grinding may not be harmful to the teeth. This should encourage orthodontists to remodel teeth by grinding to improve the final occlusal and esthetic results in a number of different cases.


The irregularity index: a quantitative score of mandibular anterior alignment. A quantitative method of assessing mandibular anterior irregularity is proposed. The technique involves measurement directly from the mandibular cast with a caliper (calibrated to at least tenths of a millimeter) held parallel to the occlusal plane. The linear displacement of the adjacent anatomic contact points of the mandibular incisors is determined, the sum of the five measurements representing the Irregularity Index value of the case. Reliability and validity of the method were tested, with favorable results. At the University of Washington, several clinical studies have been and are continuing to be performed, using this technique as one of several methods of assessing pretreatment status and posttreatment change. It is hoped that this article will aid the reader in understanding the rationale and utility of a simple quantitative tool which could be used in malocclusion assessment.


New perspectives on orientation and their benefits to clinical orthodontics - part I. The historical development of cranial reference lines, basic to cephalometric orientation, was discussed. The evolution to, away from, and back again to the Frankfort horizontal plane for descriptive and communicative purposes as benefits to the clinical orthodontist was explained. Objectives were set forth as a basis for selecting the most appropriate method in orienting the headplate for measurement and assessment. The major functions of cephalometrics were pointed out to be its use for description or classification of the face together with the malocclusion and for superimpositioning for longitudinal comparison. Each of these purposes calls for particular requirements pointing to the need for combinations of planes and multisuperimposing. The statistical findings via a computer pointed to the use of the Frankfort plane as the most desirable basic reference for descriptive reasons. Composites also suggested good order from Frankfort horizontal when serial tracings were compared. It was emphasized that true porion was used, not the ear rod of the cephalometer. For growth analysis the basion-nasion plane was shown to be longitudinally of superior value for chin behavior.


Activity of temporal and masseter muscles in children with a lateral forced bite. The activity of the temporal and masseter muscles with the mandible in postural position and during chewing, maximal bite in the intercuspal position, and swallowing were recorded electromyographically in nineteen children with laterally forced bite. Of the children, aged 8-12 years, sixteen had unilateral crossbite. In all of the individuals there was lateral deviation between the retruded contact position and the intercuspal position. The magnitude of the lateral deviation was measured with a modified gnathothesiometer. The muscle activity was recorded bilaterally from the temporal muscle and from the masseter muscle. In the postural position asymmetric activity was found in the temporal muscle suggesting that the mandible in postural position was still displaced to the side of forced bite. During chewing the activity was asymmetric both in the anterior and the posterior temporal portions. This asymmetric muscle activity was interpreted as an adaptation to avoid cuspal interferences. Also in maximal bite the muscle activity was asymmetric, while the activity during swallowing was affected less than in the other functions. The swallowing activity was, however, less in children with a forced bite than in children with normal occlusion.


Comparative analysis of changes resulting from bite plate therapy and Begg treatment. This study was based on a cephalometric study of 82 children composed of three groups: the first group treated with a flat anterior bite plate, the second group subject to the Begg therapy, and an untreated control group. The object of this study was to investigate changes in the dentofacial complex which occur as a result of bite plate therapy compared with Begg treatment. It was found that the short term changes as a result of bite plate therapy were associated with an increase in the height of the upper and lower molars. The long term changes as a result of bite plate therapy were associated with a significant increase in the height of the molars and some increase in lower incisor height. Lower facial height showed a significant increase in dimension. During Begg treatment it was found that short term changes in the dentofacial complex were associated with a significant increase in the height of the lower molars and a significant reduction in the height of the lower incisors. The long term changes of Begg therapy were associated with a significant increase in the height of the lower molars and some increase in lower facial height.


Soft-hard tissue correlations and computer drawings for the frontal view. Hard and soft tissue measurements were obtained for nine Caucasian women. Based on these measurements, regression formulas were derived to approximate the soft tissue covering based on hard tissue data. The results are presented iconically by computer drawings. The preliminary data presented would suggest a definite predictable influence exerted by the facial skeleton on the position of landmarks in the overlying soft tissue.


Multiple extraction patterns in severe discrepancy cases. Thirty-five cases have been collected from colleagues which illustrate that removal of additional maxillary teeth, following first bicuspid extractions, can allow the successful resolution of difficult discrepancy and anchorage cases. Charts 1 and 2 describe the amounts of space that might be expected by removal of additional upper bicuspids, upper first molars, and upper second molars. The findings on upper second molars are admittedly limited. Anchorage values as expressed by an efficiency percentage were approximately what would be expected from a study of anchorage values of the roots of teeth. The removal of upper second bicuspids has a better anchorage efficiency potential than the upper first molar, but this may be overcome somewhat by the greater size of the molar. Clear guidance cannot be given as to which teeth to remove in a specific case, but it is the observation of the author that for cases that are still in full Class II following four bicuspid space closure, upper second bicuspid removal would be more helpful from an anchorage perspective, whereas for cases that are in end-to-end molar relationship or require only a few millimeters to move into Class I, the upper first molar might be the tooth of choice. Also, the supper first molar removal allows for a more "normal" appearing arch assuming normal alignment and size of the upper second and third molars. The comparison with the nonextraction control group showed an enormous difference in the amount of incisor retraction that extractions provide when related to the maxilla. The nonextraction control group, though experiencing dramatic correction of Class II relationships, showed no incisor movement within the maxilla. Some problems which appeared in the sample were described. Removal of upper teeth in addition to the four first bicuspids can be a solution to an occasional anchorage, skeletal, growth or cooperation problem.


Follow-up respiratory function of a patient treated with a membrane lung. A patient in remission from acute lymphoblastic leukemia developed the adult respiratory distress syndrome. Despite "optimal respiratory therapy" and ventilatory assistance with 100 per cent inspired O2 (fraction of inspired O2 = 1) and 15 cm of positive end-expiratory pressure, the patient's arterial PO2 remained at 33 mm Hg. Prepulmonary venovenous respiratory support was given for 10 days with a spiral coil membrane lung. The data document the sequential improvement in respiratory function and suggest that despite the severe lung damage that may occur during the adult respiratory distress syndrome treated with prolonged high inspired concentration of O2, if the patient's lungs recover from the acute process, the respiratory system can recover so that there is little, if any, permanent restriction of the subject's activities as a result of respiratory impairment.


Synthesis and reaction of some 6-substituted pyrimidine nucleosides. Treatment of 5-bromouridines with sodium cyanide afforded 6-cyanouridines. Further treatment of the 6-cyano derivatives with cyanide ion at higher temperature gave the 5-cyanouridines. The 6-cyano group was transformed to thioamido, carboxy, chloromethyl cyanomethyl, carboxymethyl, and methyl group, respectively. 5-Bromocytidines were also converted to the 6-cyanocytidines, from which cytidine 6-carboxylic acid was derived. The cyano group of 6-cyanocytidines has a leaving ability toward nucleophilic substitution reactions. The circular dichroic spectra of various 6-substituted pyrimidine nucleosides showed positive Cotton effect at B2u region, which were in contrast to the reported negative bands in 6-methyl-uridine or-cytidine.