Flagellar mutants of Chlamydomonas: studies of radial spoke-defective strains by dikaryon and revertant analysis. The motility mutant of Chlamydomonas reinhardtii pf14 lacks radial spoke structures in its flagellar axonemes, and 12 proteins present in wild type are missing from a two-dimensional map (isoelectrofocusing/sodium dodecyl sulfate electrophoresis) of its (35)S-labeled flagellar proteins. Six of these same proteins are missing in pf1, which lacks spoke-heads. To determine whether any of the missing proteins represent the mutant gene product two experimental approaches have been applied. The first makes use of the fact that gametes of either mutant strain when fused with wild-type gametes to form quadriflagellate dikaryons undergo recovery of flagellar function. Recovery at the molecular level was monitored by prelabeling the mutant proteins with (35)S and allowing recovery to occur in the absence of protein synthesis. It is to be expected that the mutant gene product would not be restored as a radioactive protein and that recovery would depend on the assembly of the wild-type counterpart that is not labeled. The second technique makes use of revertants induced by UV irradiation. Dikaryon rescue in the case of pf14 leads to restoration of 11 radioactive components; only protein 3 fails to appear as a radioactive spot. For pf1 only two radioactive proteins are restored; proteins 4, 6, 9, and 10 were not radioactive. Analysis of revertants of pf1 gave evidence (altered map positions) that protein 4 is the mutant gene product. In the case of pf14, analysis of 22 revertants has not provided similar positive evidence that protein 3 is the gene product.


Addition of colchicine--tubulin complex to microtubule ends: the mechanism of substoichiometric colchicine poisoning. Colchicine blocks microtubule polymerization by an unusual substoichiometric poisoning mechanism. We have investigated the mechanism by which this poisoning occurs with several experimental approaches, and have found that colchicine acts by addition to microtubule ends as a colchicine-tubulin complex.


Parental investment, mate choice, and mate quality. Current theory in sexual selection is extended to predict within-sex variability with regard to selectivity towards mates in different mating systems. Generally, the sex that invests more in the care of each offspring should be more selective of mates than the sex investing less. Within each sex, individuals of low male quality should be less selective than individuals of high quality, but there should be less variation in selectivity among individuals of the sex investing more. When only one sex contributes parental care, however, individuals of that sex should be uniformly selective, while the other sex is expected to mate indiscriminately. Using feral pigeons (Columba livia), these hypotheses are tested for the case in which both sexes contribute substantial parental care, but in which females contribute more than males. As predicted, females were found to be more selective of mates than males were. On certain criteria, males of lower quality were less selective of mates than males of higher quality.


Transfer of codominant markers by isolated metaphase chromosomes in Chinese hamster ovary cells. Codominant mutations to methotrexate and ouabain resistance in Chinese hamster ovary cells can be transferred to recipient Chinese hamster ovary cells by isolated metaphase chromosomes. For methotrexate, both the structural change and the increased activity of dihydrofolate reductase (5,6,7,8-tetrahydrofolate:NADP+ oxidoreductase EC 1.5.1.3), characteristic of the donor cells, are observed in the transferents (cells that carry and express functions derived by chromosome transfer). The transferents are unstable in the absence of selection although stable clones can be isolated. From results obtained by fractionation of chromosomes and transfer to recipients, the methotrexate and ouabain markers can be assigned to the middle and large size-classes of chromosomes, respectively. By fractionation and transfer of chromosomes, from transferents to new recipients, evidence has been obtained that chromosome integration is not restricted to a particular chromosomal site in the recipient.


Theories of quasi-linkage and "affinity": some implications for population structure. "Quasi-linkage" refers to nonrandom assortment of genes located on different chromosomes. Although this phenomenon has been widely observed in many organisms since the early part of this century, it is barely known. Interest in it was recently rekindled by the report of an association between quasi-linkage and the expression of genes belonging to a group of cancer viruses whose genomes are integrated in mouse chromosomes. This prompted an examination of the question whether "sustained meiotic affinity," which is one of the explanations proposed for quasi-linkage, can influence population structure in a manner unattainable by other known modes of heredity. It is shown for a two-locus two-allele system that equilibrium is attained with the gametic phase disequilibrium D greater than 0, leading to a permanent excess of the preferred genotypes. The possible relationship of these concepts to the inheritance of susceptibility to cancer and other diseases is discussed.


Mutation frequencies in female mice and the estimation of genetic hazards of radiation in women. The female germ cell stage of primary importance in radiation genetic hazards is the immature, arrested oocyte. In the mouse, this stage has a near zero or zero sensitivity to mutation induction by radiation. However, the application of these mouse results to women has been questioned on the ground that the mouse arrested oocytes are highly sensitive to killing by radiation, while the human cells are not; and, furthermore, that the mature and maturing oocytes in the mouse, which are resistant to killing, are sensitive to mutation induction. The present results have a 2-fold bearing on this problem. First, a more detailed analysis of oocyte-stage sensitivity to killing and mutation induction shows that there is no consistent correlation, either negative or positive, between the two. This indicates that the sensitivity to cell killing of the mouse immature oocyte may not be sufficient reason to prevent its use in predicting the mutational response of the human immature oocyte. Second, if the much more cautious assumption is made that the human arrested oocyte might be as mutationally sensitive as the most sensitive of all oocyte stages in the mouse, namely the maturing and mature ones, then the present data on the duration of these stages permit more accurate estimates than were heretofore possible on the mutational response of these stages to chronic irradiation.


Hybridization properties of immunoglobulin mRNA: failure to detect covalently associated IgG mRNA transcripts of reiterated and unique mouse DNA. Messenger RNAs for antibody heavy (gamma) and light (kappa) chains were isolated from the polysomes of an IgG-producing mouse myeloma cell line. Polysomes engaged in heavy or light chain synthesis were separated by immunoprecipitation using rabbit antibodies specific for the mouse IgG formed. The mRNAs obtained, more than 85% specifying IgG [Legler, M. & Cohen, E. P. (1976) Biochemistry 15, 4390-4399], were used as "probes" in hybridization experiments with sheared mouse liver DNA. To determine whether mRNAs for Ig heavy and light chains contained covalently bound transcripts of unique and reiterated DNA, hybrids were isolated with or without treatment with ribonuclease (RNase) prior to fractionation and the apparent rates of hybridization were compared. A monophasic C(o)t (DNA concentration x incubation time) curve with a C(0)t(1/2) of 4000 moles of nucleotide per liter x sec, corresponding to less than five hybridization sites per haploid genome, was obtained whether or not RNase was used in the isolation protocol. With a similar experimental design, the apparent hybridization rates of heterogeneous nuclear RNA from the same cell source were clearly different. The "stringency" of the reaction was reduced by incubating the hybridization mixtures at lower temperatures in a further attempt to detect a large class of repetitive sequences that would form hybrids with the IgG mRNA used, if such sequences were present. The results, however, were the same; i.e., the apparent rates of hybridization of mRNAs for mouse antibody gamma and kappa chains with sheared mouse liver DNA were essentially the same whether or not RNase was used in the isolation procedure. Reiteration of genes in mouse liver DNA for mouse IgG could not be detected.


Zinc-binding ligands in milk and intestine: a role in neonatal nutrition? The hypothesis that a zinc-binding ligand (ZBL) recently discovered in human milk but absent from cow's milk might be related to zinc nutrition in the neonate was investigated. The zinc-binding characteristics of rat milk were examined to determine if the rat was a suitable model. By gel filtration, rat milk was found to contain a ZBL with characteristics similar to those of the ZBL found in human milk. A similar ZBL was identified in the intestinal mucosa of rats 16 days of age and older but was absent in rats from birth to 16 days. These results support the hypothesis that the ZBL of maternal milk may enhance zinc transport in the neonatal period before the development of intestinal mechanisms for zinc absorption.


Fatty liver induced in Zucker "fatty" (ff) rats by a semisynthetic diet rich in sucrose. A new fatty liver model is described in which heredity plays a major role. A marked fatty liver develops when the homozygous mutant Zucker rat (ff or "fatty") is fed a semi-synthetic diet enriched in sucrose. Lean littermates do not develop fatty livers on this experimental diet. Even old ff rats do not develop fatty livers on chow; the experimental diet is required. The fatty liver is described, including light microscopic and preliminary electron microscopic observations.


Enzyme replacement therapy in Gaucher's disease: large-scale purification of glucocerebrosidase suitable for human administration. Enzyme replacement therapy for the alleviation of Gaucher's disease has been impeded because of the difficulty in preparing large amounts of glucocerebrosidase, the enzyme that is deficient in patients with this disorder. A large-scale procedure for the purification of human placental glucocerebrosidase has been developed. The method uses cholate extraction, ammonium sulfate fractionation, acid precipitation, butanol extraction, and hydrophobic chromatography; the final enzyme preparation has a specific activity of more than 10(6) units/mg of protein with an overall recovery of 30%. In addition, the contamination of enzyme preparations, intended for human infusion, prepared by isolation procedures involving concanavalin A columns has been studied and is reported here.


Localization of motor neuron pools supplying identified muscles in normal and supernumerary legs of chick embryo. Neuromuscular specificity has been investigated in chick embryos with a grafted supernumerary leg. The nerves of the lumbar plexus are divided between the two legs so that rostral nerves innervate the grafted leg and the caudal nerves supply the host's original leg. The basic topographic organization of the histologically definable motor neuron clusters of the lateral motor columns remains unchanged by the addition of a supernumerary leg. Intramuscular injections of identified leg muscles have been used to map the intraspinal location of specific motor pools in stage 38 (12-day) embryos. In the normal embryo, the gastrocnemius muscle is innervated by neurons in a central dorsal cluster of motor neurons in segments 26-29. In six experimental cases, the motor neurons supplying the gastrocnemius muscle of a rostrally placed grafted leg were consistently located in a specific medial cluster of neurons in segments 23-25. Motor neurons in this location never normally innervate a gastrocnemius muscle, even in the very young embryos during the period of naturally occurring cell death. This observation of a systematic mismatch between a particular motor cluster and an abnormally innervated muscle indicates the operation of a selective developmental process. A hierarchy of selective chemoaffinities may best explain our experimental results.


Conformational requirements for dopamine-induced vasodilation. The availability of a series of semi-rigid analogs of dopamine and epinine has made it possible to investigate the conformational requirements for action on dopamine and beta2-adrenergic vascular receptors. The analogs were screened for dopamine-agonist action by intra-arterial injections into the renal vascular bed and for beta2-adrenergic activity by similar injections into the femoral vascular bed in dogs pretreated with phenoxybenzamine. 2-Amino-6,7-dihydroxy-1,2,3,4-tetrahydronaphthalene (A-6,7-DTN) and its N-methyl derivative (analogous to the trans beta rotamer of dopamine) exhibited pronounced dopamine-agonist activity and minimal beta2-adrenergic activity. In contrast, the semi-rigid analog of the trans alpha rotamer of dopamine (2-amino-5,6-dihydroxy-1,2,3,4-tetrahydronaphthalene; A-5,6-DTN) and its N-methyl derivative exerted pronounced beta2-adrenergic activity but were inactive as dopamine agonists.6,7-Dihydroxytetrahydroisoquinoline, a semi-rigid analog of the cis beta rotamer of dopamine, did not produce renal vasodilation. These results indicate that a conformation of dopamine similar to that found in A-6,7-DTN is required for dopamine-vascular activity, while the conformation found in A-5,6-DTN is preferred for interaction with beta2-adrenergic receptors.


Abnormal lithium and sodium transport in erythrocytes of a manic patient and some members of his family. This paper compares the transport of Li(+) and Na(+) in erythrocytes from a patient with mania and from members of his family to that in erythrocytes from normal humans. In normal human erythrocytes, Li(+) is transported by at least three operationally distinct pathways: one inhibited by ouabain (ouabain-sensitive), one by phloretin (phloretin-sensitive), and one not inhibited by either compound (insensitive). Li(+) can be driven up its electrochemical potential gradient by an oppositely directed electrochemical potential gradient for Na(+)-i.e., Li(+)/Na(+) counterflow can occur-through the phloretin-sensitive pathway but not through the other two pathways. Because ouabain-sensitive Li(+) transport is negligible under physiological conditions, Li(+) distribution between erythrocytes and plasma in vivo depends mainly on the balance between Li(+)/Na(+) counterflow and the insensitive pathway(s) of Li(+) transport. The steady-state ratio of Li(+) concentration in the erythrocytes to that in the plasma of the patient was between 2 and 3 times higher than the comparable ratio in normal persons. The phloretin-sensitive Li(+)/Na(+) counterflow system was almost absent in the erythrocytes of the patient. Furthermore, unlike those from normal individuals, the patient's erythrocytes showed no external Li(+)-stimulated, phloretin-sensitive, ouabain-insensitive Na(+) efflux. The magnitudes of the ouabain-sensitive and insensitive pathways for Li(+) transport in the patient's erythrocytes were within normal limits. The decreased Li(+)/Na(+) counterflow in the patient's erythrocytes was probably not due to the presence of an inhibitor in the plasma of the patient but rather to an intrinsic defect in the erythrocytes. Because the father and several siblings of the patient showed a similar abnormality in erythrocyte Li(+)/Na(+) transport, it is probable that this defect is inherited.


Calcium translocation by Golgi and lateral-basal membrane vesicles from rat intestine: decrease in vitamin D-deficient rats. Intestinal Ca2+ transport was studied in membrane vesicles isolated from microvillus, Golgi, and lateral-basal membrane preparations. Ca2+ uptake by these vesicles was measured by determination of 45Ca2+ associated with these membranes after collection by micropore filtration. Golgi membranes showed the highest initial rate and equilibration level of Ca2+ uptake. Approximately 90% of this Ca2+ uptake was into an osmotically responsive space, suggesting that what was measured was predominantly Ca2+ translocation. Vitamin D-deficient rats showed a markedly diminished rate of uptake and level of equilibration. These data indicate that a Ca2+-translocating process was associated with Golgi membranes to a greater extent than with surface membranes and that this process was markedly decreased in vitamin D-deficient rats. The results suggest that the Golgi apparatus participates in intestinal Ca2+ absorption.


Theoretical study of the effect of enzyme-enzyme interactions on steady-state enzyme kinetics. Equilibrium statistical mechanics is much concerned with problems involving intermolecularinteractions, either in lattices or in pure fluids or solutions. The possibility of enzyme-enzyme interactions suggests that the same problems might be studied profitably at steady state as well as at equilibrium. In the systems we consider, each of the identical enzyme molecules of the system undergoes steady-state stochastic cycling among states i equal 1,....,n. But the molecules do not cycle independently. Two neghboring molecules, in states i and j, interact with a free energy wij (a function of the distance r in the solution case). The instantaneous transition probabilities between states for a given molecule will depend on the instantaneous interactions between the molecule in question and its neighbors. The primary question of interest is how the enzyme flux is influenced by the interactions. The general problem is outlined here and some simple special cases are treated. The discussion will be continued in a following paper [Hill, T. L. (1977) Proc. Natl. Acad. Sci. USA 74, in press]


Evidence for pro-beta-nerve growth factor, a biosynthetic precursor to beta-nerve growth factor. The biosynthesis of beta-nerve growth factor (betaNGF) was studied in mouse submaxillary glands incubated with L-[(35)S]cystine. betaNGF was isolated from tissue extracts by the addition of antiserum against betaNGF and the washed immunoprecipitates were analyzed by sodium dodecyl sulfate gel electrophoresis. With short labeling periods (10 and 25 min) there is a major labeled species with an apparent molecular weight of 22,000 and a smaller peak comigrating with purified betaNGF chains (13,260). As time proceeds, the radioactivity in the 22,000 molecular weight peak plateaus, while the label in betaNGF continues to increase, until by 4 hr it greatly exceeds the radioactivity of the 22,000 molecular weight species. When glands incubated for 10 min are transferred to medium containing a large excess of unlabeled L-cystine, the 22,000 molecular weight peak gradually declines, and there is a corresponding increase in radioactivity at the betaNGF position. The 22,000 molecular weight species isolated from sodium dodecyl sulfate gels possesses all the cystine-containing peptides of betaNGF, and possibly two additional ones. When immunoprecipitates from submaxillary glands labeled for 25 min are incubated with the gamma subunit (a specific arginyl-esteropeptidase associated with betaNGF in the 7S NGF complex), the radioactivity in the 22,000 molecular weight species is converted to the betaNGF position. The results suggest that the 22,000 molecular weight species is a biosynthetic precursor to betaNGF, and that the gamma subunit may function as a specific protease in the processing event.


DNA packaging and the pathway of bacteriophage T4 head assembly. A cold-sensitive mutation in the structural gene for a minor phage T4 capsid protein (p20) leads to formation of heads containing p20 and cleaved head proteins and empty of DNA. Such heads can be filled with DNA and converted to active phages in vivo uponshift to high temperature. It appears that p20 has two distinct roles in head assembly: first, in construction of the prehead shell (blocked by ts and am mutation) and, second,in DNA packaging (blocked by cs mutation). The latter function is closely associated with gene 17 product, previously known to be required for DNA packagaing. Temperature shift studies of cs-ts double mutants and other observations allow determination of phage function required for DNA packaging. Contrary to previous proposals, we find that T4 DNA packaging is not directly coupled to and can follow DNA synthesis, protein cleavage, prehead core removal, and gene 21-mediated cleavage-induced increase in head volume. Our evidence suggests that an altered head assembly pathway exists and that DNA packaging is probably initiated by DNA-capsid (p20) interaction.


Isolation of an electrogenic K+/Ca2+ ionophore from an ionophoroprotein of beef heart mitochondria. A K+/Ca2+ electrogenic ionophore has been isolated from an ionophoroprotein of beef heart mitochondria and identified as a neutral peptide of molecular weight 1600. The amino acid composition and cationic specificity of the ionophore have been determined. The free ionophore was released from the ionophoroprotein as a consequence of tryptic digestion. The ionophoroprotein can be converted to an ionophoro peptide (molecular weight 5,100) by proteolysis without release of the free ionophore. The isolation of a K+/Ca2+ ionophore thus provides an introduction to the general technology of extracting ionophoro proteins and ofreleasing ionophores from these proteins by proteolytic digestion.


Antibodies to estrogen receptor: immunochemical similarity of estrophilin from various mammalian species. Immunoglobulin obtained from the serum of rabbits immunized with a highly purified preparation of estradiol-receptor complex from calf uterine nuclei has been shown to contain specific antibodies to the receptor protein (estrophilin) by four criteria: (a) precipitation of the radioactive steroid upon addition of goat antibody against rabbit gamma globulin to a mixture of the tritiated estradiol-receptor complex and the immunoglobulin, (b) adsorption of the estradiol-receptor complex by the immunoglobulin linked to Sepharose, (c) adsorption of the estradiol-receptor complex in the presence of the immunoglobulin by Staphylococcus aureus protein-A linked to Sepharose, and (d) the ability of the immunoglobulin to increase the sedimentation rate of the estradiol-receptor complex. Antibodies to calf nuclear estrophilin were shown to crossreact with the nuclear receptor of rat uterus, as well as with the extranuclear receptor of calf, rat, mouse, and guinea pig uterus and of human breast cancer. The antibodies do not react with either the nuclear or extranuclear dihydrotestosterone-receptor complexes of rat prostate or with the extranuclear progesterone-receptor complex of chick oviduct. These findings indicate an immunochemical similarity among estrophilins from several mammalian species, as well as between extranuclear and nuclear forms of the receptor, but not among receptor proteins for different steroid hormones.


Role of ribothymidine in mammalian tRNAPhe. We have previously reported that mammalian tRNAsPhe from variation tissues contain different amounts of ribothymidine and that a uridine methylase from Escherichia coli can quantitatively convert these tRNAs to species that contain their full complement ofribothymidine at position 23 from the 3' terminus. The role of ribothymidine in mammalian tRNAs has now been investigated by studying the ability of several highly purified mammalian tRNAsPhe, differing only in their ribothymidine content, to support poly(U)-directed poly(Phe) synthesis under various conditions. Our results indicate that the ribothymidine content of mammalian tRNAPhe can be correlated with the ability of these tRNAs to functionin vitro in a low-magnesium (6 mM), ribosome wash factor-dependent, poly(U)-directed poly(Phe) synthesis system from rat liver. Specifically, the effect of increasing the ribothymidine content in a class C mammalian tRNA becomes manifest in an increased apparent maximum velocity for the overall synthesis of poly(Phe), while the apparent Michaelis constant (Km) remains essentially unchanged. It is postulated that the modifiednucleoside ribothymidine might be involved in the regulation of protein synthesis at the level of translation in mammalian liver.


Primary structure of the membranous segment of cytochrome b5. The primary structure of the membranous segment of porcine liver microsomal cytochrome b5 has been determined. This polypeptide is at the COOH terminus of the cytochrome molecule and consists of 43 amino acids. It is essential for the insertion of the cytochrome into the endoplasmic reticular membrane. Automated sequence analysis of tryptic and cyanogen bromide/anhydrous heptafluorobutyric acid peptides provided data from which the following unique amino acid sequence was deduced: Ile-Ala-Lys-Pro-Ser-Glu-Thr-Leu-Ile-Thr-Thr-Val-Glu-Ser-Asn-Ser-Ser-Trp-Trp-Thr-Asn-Trp-Val-Ile-Pro-Ala-Ile-Ser-Ala-Leu-Val-Val-Ser-Leu-Met-Tyr-His-Phe-Tyr-Thr-Ser-Glu-Asn. A prediction of alpha-helices, beta-structures, and beta-turns basedon the sequence of this polypeptide is also presented.


Animal cell mutants defective in sterol metabolism: a specific selection procedure and partial characterization of defects. By using a chemically defined medium, a general and highly specific procedure was devised to select for mutant cells with less abundant or structurally altered sterol in their surface membranes. Within a certain concentration range, the polyene antibiotic filipin was shown to kill only cells with normal (as opposed to decreased) membrane sterol levels. Sterol-requiring derivatives of LM cells were isolated by chemical mutagenesis, filipin treatment, and cloning followed by replica plating in soft agar. Mutants (S1 and S2) are described which, when compared to normal cells, show decreased synthesis of demosterol in vivo from acetate and mevalonate relative to cell number or to fatty acid synthesis. When exogenous sterol is supplied, mutants S1 and S2 grow normally in suspension culture. However, when deprived of sterol supplement, mutant S1 grows slower than wild type cells and mutant S2 lyses within one to two generations. Gas/liquid chromatography revealed that the mutants contained a normal spectrum of fatty acids including unsaturated fatty acyl groups but, unlike wildtype cells, they have less abundant (mutant S1) or no (mutant S2) desmosterol in either the presence or absence of exogenous cholesterol. In vitro experiments with mevalonate as the substrate suggest that the defect in both mutants is in a demethylation reaction subsequent to lanosterol synthesis. The selection method developed here may permit the isolation of mutants with defective membrane incorporation of sterols and other polyisoprenoids as well as defective synthesis of these compounds.


Antipain and leupeptin restrict uterine DNA synthesis and function in mice. As in rats, administration of estradiol to ovariectomized mice results in a trypsin-like proteolytic activity in the uterus. After fractionation of uteri from estradiol-treated ovariectomized mice the protease activity was found in the 12,000 times g pellet and the nucleus, appearing first in the former. Further fractionation of the pellet by discontinuous sucrose gradient centrigugation resulted in sedimentation of the protease with 5'-nucleotidase, a marker enzyme for plasma membrane and separate from mitochondrial and lysosomal enzyme markers. Solubilization was best accomplished by lysis at 37 degrees. The soluble enzyme from mouse uterus had optimal activity at about 43 degrees and pH 8.3 and was inhibited by diisopropylfluorophosphate, tosylarginine methyl ester, antipain, and leupeptin, but not by soybean trypsin inhibitor. Inhibition in vitro by antipain and leupeptin, two low molecular weight peptides, prompted the study of their effect in vivo on the mouse uterus. After intact, cycling female mice received subcutaneous injections of antipain and leupeptin for 16 days, their uteri showed significant diminution in weight and total DNA when compared to untreated controls. Fertility rates were also diminished. Trypsin-like protease activity may be essential to normal uterine metabolism and function.


5'-Terminal capping of RNA by guanylyltransferase from HeLa cell nuclei. A soluble extract prepared from HeLa cell nuclei has been shown to catalyze the 5'-terminal modification of RNA and synthetic polyribonucleotides to form m7G(5')pppA-and m7G(5')-pppG- structures referred to as caps. The reaction involves the transfer of a GMP moiety from GTP to the 5' end of an RNA molecule containing at least two terminal phosphates. Significantly, neither the beta nor the gamma phosphates of GTP are transferred and polynucleotides with no 5'-terminal phosphate or only one are not acceptors. In the absence of methyl donor, G(5')pppA- and G(5')pppG- structures were synthesized, indicating that methylation is not required for guanylylation. Cap formation was considered to occur by the following mechanism:  (see article), in which AdoMet is S-adenosylmethionine, AdoHcy is S-adenosylhomocysteine, and (p)ppN- represents either the original 5' end of an RNA molecule or an internal site to which one or more phosphates were added after processing.


Transfer of mannose from mannosyl retinyl phosphate to protein. Upon incubation of [14C]mannose-labeled mannosyl retinyl phosphate with a membrane fraction from rat liver, mannose was transferred to an endogenous acceptor precipitable withchloroform/methanol to the extent of about 7%. The reaction proceeded linearly with time for 120 min at a pH optimum of about 7.0. The acceptor thus labeled with mannose could be solubilized by sodium dodecyl sulfate/mercaptoethanol. More than half of this acceptor appeared in the void volume of a Sephadex G-100 column. When it was digested with Pronase, a substantial proportion of it appeared between the void and bed volumes of a Sephadex G-100 column, thus indicating that it was a glycopeptide. In high-voltage paper electrophoresis, this glycopeptide moved to the cathode at low pH and to the anode at highpH. When digested with highly purified jack bean alpha-mannosidase, the glycopeptide released almost 50% of its radioactivity as mannose. That this transfer of mannose to glycoprotein from mannosyl retinyl phosphate does not take place via dolichyl mannosyl phosphate was shown by the fact that it is Mn2+ and Mg2+ independent, it is not inhibited by the presence of a 10-fold molar excess of nonradioactive GDP-mannose, and neither 14C-labeled dolichyl mannosyl phosphate nor 14-C labeled lipid pyrophosphoryl oligosaccharide could be detected during the incubation.


3-Nitropropionate, the toxic substance of Indigofera, is a suicide inactivator of succinate dehydrogenase. We have shown that 3-nitropropionate, an isoelectronic analogue of succinate, is a suicide inactivator of succinate dehydrogenase [succinate:(acceptor) oxidoreductase, EC 1.3.99.1] as follows. (i) When rat liver mitochondria oxidize succinate in the presence of 3-nitropropionate carbanion, the rate of O(2) consumption decreases exponentially to a zero value. This pattern is duplicated by subsequent additions of mitochondria. The dependence of the apparent first-order rate constant for enzyme inhibition, as well as the number of enzyme turnovers completed before inhibition, on the concentrations of 3-nitropropionate carbanion and succinate are those expected for an active site-directed and irreversible inhibitor. (ii) The inactivated enzyme is not resuscitated by centrifugation and washing of the mitochondria, in contrast to malonate-treated enzyme, and malonate protects against irreversible, inhibition. (iii) The inhibitor species is 3-nitropropionate carbanion and no external nucleophile is required for inhibition. (iv) The respiratory rates, respiratory control ratios, and ADP/O ratios obtained with NAD-linked substrates are unaffected by 3-nitropropionate carbanion. These results show that 3-nitropropionate carbanion is a highly specific, time-dependent, and irreversible inhibitor of succinate dehydrogenase. By analogy with the reaction of nitroethane with D-amino acid oxidase, the data are consistent with the hypothesis that the carbanionic inhibitor forms a covalent N-5 adduct with the active site flavin. However, the precise mechanism of inactivation, as well as mechanistic extrapolations to the oxidation of succinate, must await the elucidation of the structure of the modified enzyme. We can now explain the toxicity of plants such as Indigofera endecaphylla for mammals and fowl as being due to the irreversible blockage of the Krebs cycle by 3-nitropropionate carbanion.


Arrangement of subunits in assembled histone H4 fibers. The structure of assembled histone H4 fibers has been studied by analysis of electronmicrographs, including optical diffraction. An individual fiber has the appearance of an 80-A wide ribbon, twisted at intervals of about 330 A. Thicker fibers which have been observed seem to be bundles of ribbons. In diffraction patterns from both kinds of fiber, layer lines at axial spacings of about 1/55 A-1, 1/37 A-1, and 1/27 A-1 were most consistently observed. The possible arrangements of molecules within the twisted ribbons have been deduced and are found to be fairly closely related. The ribbons appear to consist of two parallel, unstaggered rows of repeating units, which are probably H4 dimers. The similarity between the observed layer line spacings of the H4 fibers and the spacings of the maxima in x-ray diffraction patterns from whole chromatin suggests that the H4 fibers have a structure related to that of chromatin. Since homogeneous preparations of histones H2A, H2B, and H3, or any mixture of these four histones, can form similar structures, it seems likely that the basic organization of chromatin is determined by a fibrous histone core around which the DNA is wrapped.


Ultrastructural localization of cell membrane GM1 ganglioside by cholera toxin. An immunoelectron microscopic method is described for sensitive high-resolution visualization of tissuebound cholera toxin. The principle is to incubate cells or tissue sections with toxin and then to localize the bound toxin with toxin-specific peroxidase (donor:hydrogen-peroxide oxidoreductase; EC 1.11.1.7)-conjugated antibody and enzyme substrate. Thin sections are examined for electron-opaque precipitates in a transmission electron microscope. Because of the specific binding of the toxin to membrane ganglioside G(M1), the method can be used for ultrastructural localization of this ganglioside. Semiquantitative data are obtained by titration of the limiting concentration of cholera toxin producing specific precipitates. The specificity of the method was controlled in various ways, including analyses of the correlation between the immunoelectron microscopy results and determinations of ganglioside G(M1) in tissues with different ganglioside concentrations, tissues hydrolyzed with Vibrio cholerae sialidase, tissues in which exogenous G(M1) has been incorporated, and lipid-extracted tissues. The immunoelectron microscopic method demonstrates that membrane G(M1) ganglioside is positioned on the external side exclusively. Cell-bound toxin remains in its original location on the plasma membrane surface of cells below 18 degrees , but appears to be redistributed both laterally and vertically in the membrane of cells incubated at 37 degrees for 30 min or longer. The results of this method indicate that in the central nervous system G(M1) is concentrated in the pre- and postsynaptic membranes of the synaptic terminals; a further increase in reactivity of these structures after hydrolysis of the nervous tissue with V. cholerae sialidase suggests that higher gangliosides of the same series are particularly increased in the pre- and postsynaptic junctions.


Binding of mouse interferon to polynucleotides. The polynucleotides poly(I) and poly(U), and to a lesser extent poly(G), are capable of desorbing mouse interferon from blue dextran-Sepharose columns, whereas poly(A) andpoly(C) are without effect. When covalently bound to agarose, poly(I), poly(U), and also poly(A) act as potent ligands for purification of interferon by affinity chromatography. Furthermore, poly(I) and poly(U) confer a significant degree of protection to interferon against thermal denaturation. Taken together, these observations point to a direct interaction of interferon with these polymers and suggest that interferon molecules have a polynucleotide attachment site. Possible implications for the concept of interferon induction are discussed.


Prescence of tadpole and adult globin RNA sequences in oocytes of Xenopus laevis. Complementary DNA transcribed from adult Xenopus laevis globin mRNA was used to assay ovary RNA from Xenopus for the presence of globin sequences by RNA.cDNA hybridization. These sequences are present at approximately the same concentration as the majority of poly(A)-containing ovary sequences. The sequences are also found at approximately 200,000 copies per cell in poly(A)-containing RNA extracted from mature oocytes. To rule out contamination of the oocytes with somatic cells, two additional experiments were performed. First, RNA isolated from ovulated unfertilized eggs, which are devoid of somatic cells, was also shown to contain the globin sequences. Second, globin mRNA was isolated from Xenopus tadpoles. Adult globin mRNA is free of the tadpole sequence and no homology was detected between adult and tadpoles globin RNA. The ovary was shown to contain tadpole globin RNA at nearly the same concentration as the adult sequences. Thus, the results cannot be explained by contamination with erythroid cells which should contain only the adult sequence. The swimming tadpole, which possesses an active circulatory system, was also assayed for the tadpole and adult globin sequences. Whereas the adult sequences are present at approximately the same concentration as in the mature oocyte, the concentration of the tadpole sequences increases at least 300-fold in the first 3 days following fertilization.


Temporal sequence of hormonal interactions during the prereplicative phase of quiescent cultured 3T3 fibroblasts. After addition of prostaglandin F2alpha (PGF2alpha) and insulin to quiescent cultured 3T3 fibroblasts a constant lag phase of 15 hr occurs before an increased rate of cellular exit from G1 is observed. The latter process follows first order kinetics, which can be quantified by a rate constant k. The temporal relationship of the interactions of PGF2 alpha, insulin, and hydrocortisone to produce alterations in the rate of exit from G1 was investigated. PGF2alpha establishes a constant lag phase and produces alterations in the rate constant k. These two effects can be partially separated by adding two concentrations of PGF2alpha at different times. Insulin fails to establish the lag phase but can stimulate the effect of PGF2alpha when added at any time after PGF2alpha. Hydrocortisone reduces the value of k when added between 0 and 3 hr after PGF2alpha. These results show that the lag phase can beseparated into temporal regions during which hormones can interact to produce changes in the rate of cellular exit from G1.


Succinyl concanavalin A stimulates and antimicrotubular drugs inhibit the synthesis of a brain-specific protein in rat glial cells. The relative synthesis of the brain-specific s100 protein increased as clonal rat glial cells, C6, progressed from logarithmic to stationary growth in monolayer culture. Drugs that disrupt microtubules, such as colchicine, vinblastine, Colcemid, and podophyllotoxin, inhibited the relative synthesis of S100 protein in stationary cultures. Colchicine (0.1 muM) caused a 50% inhibition of the relative synthesis of S100 protein whereas lumicolchicine,an isomer of colchicine that does not disrupt the microtubular system, had no effect. Succinylated concanavalin A (500 mug/ml) increased relative synthesis in logarithmic but not stationary cultures. These results suggest that signals inducing an increase in the relative synthesis of S100 protein in stationary cultures are transmitted intracellularly from the cell membrane by the microtubular network.


Separation and infectivity of circular and linear forms of potato spindle tuber viroid. Potato spindle tuber viroid can be separated into two fractions by polyacrylamide gelelectrophoresis in the presence of formamide and urea. One fraction contains predominantly circular molecules; the second fraction contains almost exclusively linear molecules. The purity of the fractions was estimated by electron microscopy of formaldehyde-denatured molecules. The length distributions of denatured circular and linear molecules were determined. Both circular and linear molecules were found to be infectious.


Low-molecular-weight peptide inhibits RNA synthesis in human leukemic and phytohemagglutinin-stimulated leukocytes and globin mRNA transcription in differentiating Friend cells. The RNA synthesis of human leukemic leukocytes and phytohemagglutinin-stimulated lymphocytes is markedly reduced by administration of a low-molecular-weight nonhistone peptide factor from calf thymus. Treatment with the factor strongly inhibits hemoglobin production and globin mRNA transcription in dimethyl sulfoxide-stimulated Friend cells without appreciably modifying the rate of cell growth. Evidence for specificity of these effects is provided by the lack of action of the factor on both growth rate and RNA synthesis of a number of nondifferentiating cell lines from various animal species. After removal of the compound, both human lymphocytes and Friend cells can be stimulated by phytohemagglutinin and by dimethyl sulfoxide, respectively, ruling out any toxic effect.


Pure and mixed erythroid colony formation in vitro stimulated by spleen conditioned medium with no detectable erythropoietin. Cells from CBA fetal mouse liver formed pure or mixed erythroid colonies in semisolid agarculture after stimulation by medium conditioned by pokeweed mitogen-stimulated mouse spleen cells. In general shape, the erythroid colonies resembled typical 7-day single or multiple (burst) colonies. However one-third to one-half contained, in addition to erythroid cells, macrophages and neutrophils and, less commonly, megakaryocytes or eosinophils. Culture of micro manipulated single colony-forming cells showed these erythroid colonies to be clones. Colony-forming cells declined in frequency with advancing fetal age, but low numbers were detectable in adult bone marrow. Assays of spleen conditioned medium in polycythemic mice failed to detect erythropoietin; the cloning system may detect a fetal type of erythropoietin-independent, erythropoietic cell since few were detected in adult marrow.


Intracellular distributions of mechanochemical proteins in cultured fibroblasts. We have used methods that have allowed simultaneous fluorescent staining of intracellular actin together with either myosin, filamin, or tubulin in normal rat kidney fibroblasts in monolayer culture. In the main portions of the cell body, the actin, myosin, and filamin are all present in two structures: in one, the three proteins are present in the same fiber bundles (stress fibers); in the other, there is a diffuse distribution of the three proteins. On portions of the cell periphery however-in the basal regions of microspikes, in ruffles, and in regions of cell-cell contact-actin and filamin are present, but myosin is severely depleted or absent. Microtubules are present in the cell body in a distribution independent of the stress fibers and are mostly absent from the cell periphery. Microspikes and ruffles are highly dynamic structures on the cell surface, and regions of cell-cell contact generally result from the association of ruffles on the two contacting cells. Therefore, the presence of filamin and actin but not myosin in these specialized regions on the cell surface, together with the recent demonstration [Wang, K. & Singer, S. J. (1977) Proc. Natl. Acad. Sci. USA 74, 2021-2025)] that pure filamin interacts with individual F-actin filaments in solution to form fiber bundles and sheet-like structures, suggest that in vivo filamin-actin interactions play an important role in the control of actin filament structure, in cell motility, and in the stabilization of cell-cell contacts.


Loss of growth control and differentiation in the fu-1 variant of the L8 line of rat myoblasts. A nonfusing variant, fu-1, of the L8 line of rat myoblasts was isolated and characterized with respect to its growth in vitro and developmental properties. Comparative analyses of density-dependent inhibition of growth, serum requirements, cell adhesiveness, colony formation in soft agar, and hexose transport in L8 and fu-1 cells support the conclusion that the fu-1 cells are transformed. In addition, fu-1, but not L8, cells promote the development of tumors in athymic nude mice. fu-1 cells also do not make increased levels of creatinekinase (ATP:creatine N-phosphotransferase, EC 2.7.3.2) or myosin and they express an endogenous type-C virus. Both L8 and fu-1 cells express myokinase (ATP:AMP phosphotransferase, EC 2.7.4.3) activities in single cells. In contrast to fu-1 cells, the parent L8 line has increased creatine kinase and myosin after fusion and spontaneously contracts; expression of an endogenous virus could not be detected in these cells. These results suggest that loss of the ability to differentiate normally is associated with the loss of the normal control of cell division of myoblasts grown in vitro and in vivo.


Morphological differentiation of neuroblastoma cells in medium supplemented with delipidated serum. Neuroblastoma cells have been found to extend neurites when grown in a medium supplemented with delipidated fetal calf serum. Fetal calf sera from different commercial sources give rise to marked differences in the degree of this spontaneous morphological differentiation. The phenomenon can be prevented by the addition of certain fatty acids; oleic acid is especially effective. The serum-free conditioned medium from glial cells can quantitatively antagonize the effect of oleic acid, suggesting that glial factor activity could be due to components [lipids and/or macromolecular factors(s)] that are able to modify the properties of the neuroblastoma cell membrane.


Dissociation of tumor promoter-stimulated ornithine decarboxylase activity and DNA synthesis in mouse epidermis in vivo and in vitro by fluocinolone acetonide, a tumor-promotion inhibitor. 12-O-Tetradecanoyl phorbol-13-acetate (TPA), a tumor promoter, stimulates DNA synthesis in mouse epidermal cells in vivo and in vitro. This response appears to be mediated through polyamine metabolism because ornithine decarboxylase (L-ornithine carboxy-lyase, EC 4.1.1.17)activity is markedly increased shortly after promoter exposure and this induction varies in magnitude according to dose and promoter potency of a series of phorbol esters. In vitro, exogenous putrescine (0.01-10 mM) results in a dose-related increase and prolongation of promoter-stimulated DNA DNA synthesis, a phenomenon noted in other systems of polyamine-mediated growth stimulation. The anti-inflammatory steroid fluocinolone acetonide (FA), an inhibitor of tumor promotion, prevents TPA stimulation of epidermal proliferation in vivo and in vitro. In vitro, FA most effectively prevents stimulation of DNA synthesis when applied is not required. Paradoxially, FA potentiates the increase in ornithine decarboxylase activity after TPA administeration both in vivo and in vitro. Furthermore, the inhibition of TPA-stimulated DNA synthesis by FA in vitro can be reversed by exogenous putrescine. These results suggestthat FA exerts its antipromotion effect by reducing the sensitivity of the cell to polyamines or by reducing intracellular polyamine levels.


Stable association of the human transgenome and host murine chromosomes demonstrated with trispecific microcell hybrids. Trispecific microcell hybrids were prepared by transferring limited numbers of chromosomes from a human/mouse gene-transfer cell line to a Chinese hamster recipient line. The donor cells employed were murine L-cells that stably expressed the human form of the enzyme hypoxanthine phosphoribosyltransferase. Karyotypic, zymographic, and back-selection tests of the resulting human/mouse/Chinese hamster microcell hybrids provided strong genetic evidence for a stable association of the human transgenome with host murine chromosomes in stable gene-transfer cell lines. This association, which may represent physical integration of the transgenome into the host cell genome, occurred at multiple chromosomal sites.


Rapid speciation and chromosomal evolution in mammals. To test the hypothesis that population subdivision into small demes promotes both rapid speciation and evolutionary changes in gene arrangement by inbreeding and drift, we estimated rates of speciation and rates of chromosomal evolution in 225 genera of vertebrates. Rates of speciation were estimated by considering the number of living species in each genus and the fossil record of each genus as well as information about extinction rates. Speciation rate was strongly correlated with rate of chromosomal evolution and average rates of speciation in lower vertebrate genera were one-fifth those in mammalian genera. Genera with high karyotypic diversity and rapid speciation rates may generally have small effective population size (Ne), whereas large Ne values may be associated with karyotypically uniform genera and slow rates of speciation. Speciation and chromosomal evolution seem fastest in those genera with species organized into clans or harems (e.g., some primates and horses) or with limited adult vagility and juvenile dispersal, patchy distribution, and strong individual territoriality (e.g., some rodents). This is consistent with the above hypothesis regarding the evolutionary importance of demes.


Overexpression of an unstably inherited gene in cultured mouse cells. The specific activity of hypoxanthine-guanine phosphoribosyltransferase (IMP:pyrophosphate phosphoribosyltransferase, EC 2.4.2.8) is increased up to 58-fold in unstable gene transferents produced by the transfer of cell-free chromosomal material from one mouse L cell line to another; the specific activity of this enzyme returns to normal levels when the transferred gene becomes stabilized. This phenomenon, which is not observed in comparable heterospecific transfers, may be an effect of gene dosage (multiple copies of the transferred genetic fragment in the unstable gene transferents), or it may represent an escape of the unstably inherited gene from the normal regulatory mechanisms of the recipient cell.


Heterogeneity of nephritic factor and its identification as an immunoglobulin. Complement C3 nephritic factor (NeF) produces alternative pathway-meciated C3 cleavage by binding to and stabilizing the alternative pathway C3 convertase, C3bBb. Some studies have suggested that NeF is an immunoglobulin, while others conclude that it is a distinct serum protein. The heterogeneity of NeF was evaluated by electrophoresis and isoelectric focusing of NeF-containing serum followed by hemolytic demonstration of NeF activity in agar gel. With each method, diffuse cathodal zones or multiple bands of hemolysis developed, which revealed remarkable variations in patterns from patient to patient. NeF activity was absorbed by and eluted from insolubilized antibody to Fc and Fab fragments of IgG. Immunoabsorption of six NeF-containing sera with insolubilized anti-kappa and anti-lambda light chain antisera revealed that NeF had kappa antigenic determinants in three, lambda antigenic determinants in one, and both kappa and lambda antigenic determinants in two. These data indicate that NeF is an oligoclonal immunoglobulin. Because NeF binds to the alternative pathway C3 convertase, C3bBb, we suggest that it is an antibody to a conformational antigen of the C3-factor B complex, and thereby stabilizes this complex.


Antibodies to tubulin in normal nonimmunized animals. Sera of normal nonimmunized rabbits, pigs, calves, and humans contain tubulin-reactive antibodies. Usually, low amounts of antibodies against tubulin of the IgG class (2.5-4 mg/100 ml of serum from nonimmunized animals) were isolated. Anti-tubulin antibodies were also produced by injecting pig tubulin in complete Freund's adjuvant into rabbits. Slightly higher amounts of anti-tubulin antibody were isolated from sera of immunized rabbits (7 mg/100 ml of serum). The cytoplasmic network of microtubules of Tcc 36 mouse cells in culture was not clearly stained by natural anti-tubulin antibodies, but dense staining of the centrosphere was observed. In contrast, induced anti-tubulin antibodies densely stained cytoplasmic microtubular networks. Vinblastine-induced tubulin paracrystals were equally stained by natural and induced anti-tubulin antibodies.


Inhibition of human platelet thromboxane synthetase by 9,11-azoprosta-5,13-dienoic acid. The synthetic prostaglandin analog 9,11-azoprosta-5,13-dienoic acid (azo analog I) has been found to be a potent inhibitor of human platelet thromboxane synthetase by three independent analytical methods: electron-capture gas chromatography, radioisotopic thin-layer chromatography, and radioimmunoassay. In the presence of azo analog I, human platelet aggregation induced by either the prostaglandin endoperoxide PGH2 or arachidonic acid was antagonized. The addition of azo analog I shifted the transformation of endoperoxides away from thromboxane synthesis and toward prostaglandin E2 synthesis. The specificity of azo analog I is demonstrated by its selective inhibition of the second wave of either ADP- or epinephrine-induced platelet aggregation. These data indicate that PGH2 must be converted to thromboxane A2 in order to induce human platelet aggregation.


Active touch exploration of extrapersonal space elicits specific electrogenesis in the right cerebral hemisphere of intact right-handed man. Language and analytic processing are currently thought to be represented in the left hemisphere, whereas spatial and holistic processing would involve primarily the right hemisphere in man. An experimental paradigm for engaging the nonlanguage hemisphere (generally the right) is described. This involves active touch exploration with the index finger to identify the orientation of a ridge with respect to the subject's body. The task is compatible with the electronic averaging of transient event-related cerebral potentials recorded from the intact scalp. A consistent positive electrogenesis of 1-5 muV and about 0.5-1.5 sec in duration was recorded over the nonlanguage hemisphere, regardless of whether the left or the right index finger performed the tactile scanning. The lateralized specific electrogenesis did not extend to the midline, and it is to be differentiated from the decision P300 component. These findings provide a new procedure for analyzing, in intact man, measurable focal potentials associated with unique processor subsystems during cognitive behavior. The method will make it possible to investigate the dynamic distribution of processing tasks between the two hemispheres in normal man in whom the commissural integration is normal, thereby adding to the data collected on patients with unilateral brain lesions or with surgical transection of the corpus callosum.


Neurophysin biosynthesis in normal rats and in rats with hereditary diabetes insipidus. When [35S]cysteine was injected adjacent to the supraoptic nucleus (SON) in rats, it was rapidly incorporated into proteins in the SON. The [35S]cysteine-labeled proteins extracted from the SON were separated by isoelectric focusing on polyacrylamide gels. Twenty minutes after the injection of [35S]cysteine, two major labeled peaks (pI = 5.4 and 6.1) were found in the SON of normal rats; Brattleboro rats had only one major labeled peak (pI = 5.4). One hour after the injection, four major radioactive peaks were found in the SON of normal animals (pI = 5.1, 5.4, 5.6, and 6.1). Animals with diabetes insipidus had only two major labeled proteins (pI = 5.1 AND 5.4). Twenty-four hours after normal rats were injected with [35S]cysteine, all of the labeled peaks described above, except for the one with pI = 5.1, had decreased markedly in size and a small amount of labeled protein with pI about 4.8 was present in the SON. After 24 hr the posterior pituitary of normal animals contained two [35S]cysteine-labeled proteins with pI = 4.6 AND 4.8. The pituitaries of Brattleboro rats had only the pI = 4.6 labeled protein. These pulse-chase data, with data we have presented elsewhere, indicate that the vasopressin- and oxytocin-neurophysins are synthesized as parts of separate precursors (pI = 6.1 and 5.4, respectively). These precursors are converted into at least two intermediates (pI = 5.6 and 5.1) which, in turn, yield the vasopressin-neurophysin (pI = 4.8) and the oxytocin-neurophysin (pI = 4.6).


Effectof MgATP on stiffness measured at two frequencies in Ca2+-activated muscle fibers. The stiffness of skinned crayfish single muscle fibers was continuously monitored at two frequencies. The length of the fibers was oscillated by the sum of two sine waves (5 Hz and 100 Hz) of small amplitudes. In saline containing saturating amounts of Ca2+, the stiffness ratio (5 Hz:100Hz) was constant as the MgATP (substrate) concentration was raised from 0 to 2 mu M, then it decreased with a further increment in MgATP. The systematic decrease in the stiffness ratio in MgATP above 2 mu M indicates the presence of faster transitions in the cross-bridge cycle. This dependence of the stiffness ratio on MgATP is predictable if we use the two-state model of A. F. Huxley (1957) with a modification, in which MgATP promotes the dissociation of the attached cross-bridges.


HLA typing of leukemic cells and lymphoblastoid cell lines: effects of non-HLA antibodies in common typing sera. Several laboratories have recently observed that common HLA typing sera contain antibodies to various nonHLA antigens, including leukemia-associated antigens and normal platelet, granulocyte, or B-lymphocyte-specific alloantigens. In order to study the possible effects of these antibodies on HLA typing data in leukemia, several of these sera were adsorbed with HLA typed platelets, purified B-lymphocytes, cultured lymphoblasts, or mononuclear cells from patients with active leukemia of various types, and retested for residual activity against previously reacting cells. The results indicate that many of the reactions of leukemia cells to HLA typing sera represent reactions to nonHLA antibodies, many of which are directed against antigens common to cultured lymphoblasts, leukemia cells of several types, and normal B-cells. In the absence of family studies or prior serum adsorption, caution should be used in interpretation of all data related to HLA antigens in leukemia.


Combined therapy of osteosarcoma: preoperative irradiation, surgery, and chemotherapy. The rationale for prebiopsy and preoperative irradiation in a multimodal approach to treatment of osteosarcoma is presented. Six patients with osteosarcoma underwent preoperative irradiation, amputation, and elective chemotherapy. Five of these also received prebiopsy irradiation: three survive without metastases at 29, 36, and 56 months, with no therapy for 10, 16, and 37 months, respectively; two patients died at 6 and 19 months of pulmonary metastases which appeared at 2 and 10 months, respectively. The latter two did not receive prebiopsy irradiation. Another patient, whose pulmonary metastasis regressed at 6 months with adriamycin and was later resected, died of cardiac failure at 59 months without evident metastasis. Immunologic aspects of the disease are also discussed.


Radiotherapy-induced rib osteosarcoma after successful treatment of lung cancer. Radiation-induced osteogenic sarcoma is recognized as a long-term complication of radiation therapy. The authors report a case of rib osteosarcoma which developed after successful treatment of adenocarcinoma of the lung.


[Surgical treatment of loss of substance of the lower lip]. Repair of loss of substance of the lower lip has been the object of numerous studies. Each author has used his own ingenuity to describe a new type of flap aimed at restoring a particular form of loss of substance. The techniques suggested here are capable of solving a majority of problems on the basis of simple and original solutions. These flaps make it possible to resolve the problems posed by most cases of loss of substance of the lower lip. The choice of these plastic procedures permits restoration which is at one and the same time functional and anesthetic.


[Cysts and fistulas of the first branchial cleft with parotid symptomatology]. On the basis of two cases, one a fistula and the other a cyst of the first brachial cleft with chiefly parotid manifestations, a review of the embryology is given which explains the anatomical features. The elements in their clinical diagnosis, complemented by fistulography and sialography are compared with operative findings, the essential steps in which are as follows: 1) Identification of the facial nerve which by necessity overlies the lesions. 2) Partial resection of the superficial lobe of the parotid. 3) The essential resection of the external auditory meatus, the keystone of radical cure of these cysts and fistulae.


[Dentofacial orthopedics and morphopsychology]. Acquired maxillo-facial deformities and in particular those related to problems of facial expression are nevertheless influenced by certain hereditary factors, amongst which the constitutional type or the temperament of the individual play a major role. Morpho-psychology (L. Corman), or the art of relating physical details (especially the shape and characteristics of the face) with psychological state demonstrates correlations between the type of deformity and that of the temperament and explains them. After a brief review of the essential principles of morpho-psychology, the authors study a number of special morpho-psychological types presenting with a significant frequency maxillo-facial deformities, themselves special, and which for this reason they call "morpho-psychological types with a high orthodontic risk". They conclude that it would appear that insufficient importance is attributed at present to psychological factors in the aetiopathogenesis of dysmorphoses and feel that an overall approach to the deformed child is indispensable to progress in the specialty.


[Chondrosarcoma of the upper jaw. Apropos of a clinical case]. Initially it has been a tendency to place chondrosarcoma in the general category of osteogenic sarcoma. Jaffe et Lichtenstein claimed that the concept of chondrosarcoma as a separate entity has a firm clinical and histological basis. Chondrosarcomas may arise in peripheral long and flat bones. The maxillary location is very rare, it represents about 0,7% of the whole body. 40 well documented instances are yet counted in the literature. Reporting a personnal case, the author proceeds to a comparative study between the three varieties of maxillary sarcomas, evokes the special histogenesis in this location, he reminds the interest of bone electrocoagulation as curative treatment, and discusses the difficulty in making a prognosis 3 years after surgery without local recurrence or metastasis in this case.


[The human head, a cognitive and communicative system. Practical applications]. The human face is a sensory complex with the task of transmitting to the central nervous system most of the information and signals from the surroundings. This perceptive centripetal activity is linked with continual centrifugal transmissions to the surroundings of a relational type. The cerebral extremity of a human being is ethological in essence. The human cephalic phenotype is, during the whole of its development, a succession of responses of the genome to the stresses of the environment: it is an open system. The phylogenesis of the cerebral cognitive organs and the phylogenesis of the skeleton which protects them are linked. The growth of the membranous cephalic skeleton is under the control of these cognitive organs. Practical applications are deduced from this in connection with the pre-operative functional test for prognathisms.


[Bone healing and the prevention of recurrence in the surgical treatment of prognathism]. The authors analyse a series of 60 cases of mandibular prognathism treated using the technique of Trauner-Obwegeser modified by Dalpont. The last 40 cases involved no surgical procedure as far as the musculature was concerned and no difference in the course of bone healing was noted.


[Psychology and dentofacial orthopedics. Apropos of various results]. What should be done about the child who sucks his thumb? If the child has no dento-maxillary deformity, he should be left to suck his thumb. In the presence of deformities, it would seem wise not to interfere before the age of 6 or 7 years. Firstly, the majority of thumb suckers abandon this behavior spontaneously between 3 and 7 years, and secondly it is only at the age of 6 years that the milk teeth begin to be replaced by the adult dentition. An early intervention has the risk of being ineffective or even of reinforcing the habit and leading to other problems. After 7 years, the child with deformities should be examined and in the absence of an psychological contraindication, "invigoration psychotherapy" should be undertaken in order to put a stop to the thumb sucking habit. Finally, when the 8 year old continues to suck his thumb despite psychotherapy by the psychologist or the stomatologist, the advice of a child psychiatrist must be sought in order to determine whether the symptom which the habit represents should be disturbed or not.


[Semeiologic meaning of dilatation of the dental canal]. The authors report two cases of dilatation of the dental canal due to metastases. They review the clinical picture of dilatations of the dental canal which, other than being related to metastases, may be secondary to a haemangioma or a neurofibroma. They express the desire that measurements may be helpful in considering cases at the limit of normal, as for the internal auditory meatus and the optic canals.


[Total reconstruction of the mandible with an endoprosthesis]. The authors present a reconstruction after total mandibulectomy, using a Kirschner wire, modelled and adjusted at the time of insertion. They emphasis the simplicity of the technique and the quality of the result.


[Oro-sinusal communications. Methods of closure]. Three surgical methods are proposed by the authors for the treatment of an oralantral bistula (1. Wassmund-Rehrmann-Schüchardt--2. Kazanjian-Schüchardt--3. J. M .Vaillant). This is an experience drawn by 450 cases treated in the Ceinic of Oral Surgery of the University of Salonica (Greece), during the last three years. Fistulas, immediately appeared after extraction of an upper caspid of premolare, surgical removal of teeth, or apex, intrasinusaly located, long persistance of a fistula because of a kyste or tumor in the upper jaw have been the subjects of the present paper.


[Streptococci isolated from the blood after tooth extraction. Their role in subacute bacterial endocarditis]. A classification, according to the Lancefield's method, of the streptocoques isolated in the blood, after teeth extractions, as well as a determination of their sensibility to the antibiotics, are attempted in this study. It concerns to 23 adults, well supported, to whom multiple extractions, under general anesthesia have been done. We studied both biochemical and serological reactions, and a test for the antibiotic resistance is realised. We insiste in the contrindiction of streptomycine and sulfanilamides in the treatment or the prevention of bacteremia of dental origine.


[Gingival metastasis of a malignant bronchial carcinoid tumor. Apropos of 1 case]. Secondary tumors of the mouth are rare and especially seated on the maxillaries. Metastasis to the soft tissues of the mouth, especially of cancers situated far from ORL and buccal areas are exceptional. The authors report an observation of a bronchial carcinoïd tumor with metastasis to the gums in a 24-year old man. The rapidity of the evolution of the tumor, the precocity and the very exceptional site of the metastasis as well as certain particularities of the histological pattern of the primitive bronchial tumor, once again states the problem of the confines with the Oat-Cell Carcinomas.


Leucocyte alkaline phosphatase elevation in human acute leukaemia derived cell lines cultured in diffusion chambers. Leucocyte alkaline phosphatase (LAP) was histochemically detected in 7 to 18% of cells in tissue culture lines derived from the peripheral blood or bone marrow of each of 5 patients with untreated acute myelogenous or monomyelogenous leukaemia and in 30% of the cells in a clonal line of a rat promyelocytic leukaemia. Following transfer to diffusion chambers intraperitoneally implanted into total body irradiated rats, LAP levels were detected in up to 92% of human and 80% of rat leucocytes. There was no associated morphologic differentiation. In rat leukaemia cells peroxidase and myeloid specific esterase also increased from tissue culture levels. Return of cells to tissue culture decreased enzymes to pre-implant levels. Addition of plasma or peritoneal fluid from irradiated rats to cells in tissue culture again induced LAP. In contrast, LAP was not increased under these conditions with cell lines derived from patients with acute lymphatic leukaemia, or Sezary cell leukaemia. These studies indicate that a humoral factor in peritoneal fluid and plasma of irradiated rats increases LAP in human as well as rat leucocytes.


Hodgkin's disease and acute leukaemia. A clinical and cytogenetic study. Three patients developing acute myeloid leukaemia after treatment for Hodgkin's disease are reported. They appeared among 153 patients treated since 1971. All three patients were treated with high-voltage radiation of large target volumes as well as multiple-drug chemotherapy. In two of the patients chromosomal analyses of the leukaemic bone marrow cells were performed by modern banding technique. Chromosome abnormalities were found in both patients. The value of cytogenetic analysis with the new staining techniques for elucidating the mechanism of leukaemogenesis in Hodgkin's disease is discussed.


Mitotic activity of the granulopoietic precursor cells in the peripheral blood in chronic myeloid leukaemia. The mitotic indices (MI) of granulopoietic precursor cells in peripheral blood and bone marrow were studied in 38 patients with typical chronic myeloid leukaemia (CML) in the chronic phase. The MI in the peripheral blood were very low, in the median 0.07%, compared to those in the bone marrow with a median of 1.07%. The blood MI were significantly increasing with raising WCC and the values of the MI above the median were combined with short survival times. The bone marrow MI were negatively correlated to the blood MI and it is suggested that this is a sign of an increased exchange of cells between bone marrow and blood.


[Therapeutic results and prognostic factors in acute lymphatic leukemia in the adult]. Induction chemotherapy consisting of vincristine, prednisone and L-asparaginase was given to 22 adult patients with acute lymphatic leukemia. Manintenance treatment consisted of methotrexate, 6-mercaptopurine, prednisone and vincristine. Of the 22 patients treated, 14 had a complete remission. The median remission duration was 14 months and the median survival 20 months. 8 further patients were included in an attempt to determine the significance of prognostic factors, but none of the parameters studied influence the course of disease with statistical significance. In general, younger patients and those with lower leukocyte counts at the time of diagnosis seemed to fare better.


[Relativity and limits of success yesterday and today]. A comparing of today's dentistry and the time limits of its success with dental medecine of yesterday, we state an unquestionable advance in the preventive field. Causes of caries and parodontopathies are so well known that proper preventive measures have shown, within the last twenty years, statistically relevant success. Such positive results may be endangered by politics which tend to eliminate personal responsibility and which establish insurance systems by which even the most expensive dental restorations must be paid by the collective community. Thus prophylactic measures become unimportant to the individual. Preventive thinking has various facets: The system of filling gaps and holes must yield to the system of rehabilitation at every level. The basis to this is a high standard of professional ethics. Its realization is easier if dentistry is not subdivided into too many specialties. On the other hand, the dentist must have a large spectrum of knowledge, which is only obtainable through continuous education. The younger generations are profiting most from this development of knowledge, but the old and the handicapped should not be forgotten in the process. It will be one of the noblest tasks of the profession henceforward to care for the treatment of this dentally underpriviledged part of our population.


[Legal aspects of failure in dentistry]. Conflicts between dentist and patient are increasing in a disquieting way. The reasons for this should be discovered. It seems indispensable to modify the attitude of the practitioner towards his patient, because the latter may have justified claims. To ignore them would mean to cause more trouble, indeed to lead water on the mill of those who would eliminate our liberal profession. Let us not forget that for many people, the practice of liberty is an insupportable challenge. It will be the attitude and psychological feeling which may enable us to avoid that state control of the whole profession.


[Cases of clinical failures].

[Cases of clinical failures]. During the past 4 years, the Department of Health of the State of Berne received 91 complaints of patients about dental treatment. Most of the cases could be settled by proposals of the dental section of the state medical council. Most frequently crown and bridgework was the reason for complaints, secondly partial and complete dentures followed by restorative work and excessive grinding on teeth. Complete information of the patient about treatment possibilities, clear estimate of costs and referals for consultations would reduce the number of complaints.


[Failures in periodontal treatments].

[Failures in periodontal treatments]. Failures in periodontal therapy may be caused by a faulty choice of patients, insufficient diagnosis, prognosis and planning, faulty choice of therapy, treatment and follow-up treatment. Particularly important is the motivation of the patient, his collaboration in periodontal therapy is decisive for success or failure of the treatment. Lasting success may only be expected if patients are regularly recalled, their brushing and cleaning habits may be checked and further treatments be dispensed in time.


[Criteria of success or failure in conservative dental treatments]. Any restorative procedure which is not effected with clinical precision and with due consideration to the principles of preventive dentistry is bound to fail. The main causes of failure, whether direct or indirect, are iatrogenic and can be avoided. Restorative procedures may be qualified as a success when a favorable, durable result is obtained. This result depends partly on the technique employed and partly on the products used, products which are too often tardively and insufficiently tested. This state of affairs underlines the urgent necessity of introducing standardized clinical controls for all new dental products.


[Failures and limitations of orthodontics in general practice]. 5 cases are presented to review the causes of orthodontic failure: ignorance of growth and development; limited technics often associated with overestimation of clinical capabilities. The concept of extracting 4 premolars followed by "what ever nature will do with it" or by a partial so-called "social treatment" (read with a minimum of appliances) can no longer be accepted.


[Long-term clinical results in fixed and removable prostheses]. The author, on the basis of 25 years of clinical experience, defines a logical and coherent system for the choice of mixed prosthetic solutions, fixed and removable. He believes in splinting and rigid attachment.


[Biological and psychological aspects of geriatric dentistry]. A normal adult becomes a normal old man when going through a whole process of physiologic involution called natural ageing, which does not spare the stomato-gnathic system. The authors first review the biological aspects of ageing as seen on the skin and facial contour, on dentition and teeth, on periodontium, oral mucosa, tongue, salivary glands, facial and masticatory muscles, facial skeleton and temporomandibular joints. They then follow with the psychological aspects of ageing, going necessarily beyond the limits of dental medicine. The authors finally discuss means to prevent that natural ageing becomes pathologic.


[Psychological aspects of geriatric dentistry]. The goal of geriatry and gerontology is not to prolong life at all cost but to make this last lap in life bearable. The dentist is often confronted with geriatric realities. The ideas on the psychology of the third age have considerably evoluated within the last ten years and more specific methods have been applied to study old age. Upon these new approaches, it seems possible to improve treatment on a preventive as well as on a therapeutic level.


[General medical concepts for the dental treatment of aging patients]. Patients, particularly older ones, with internal medical diseases, may be hazards for the dental treatment. Therefore, anamnestic data seem to be most important in order to uncover the hazard patient (diabetes, survived heart attacks, anticoagulation, rheumatic heart affections, hypertonics, allergies). Local anesthesia can be another problem, because it may lead to collapse (psychogenous or from anaphylactic shock). Focal infection, although in general overrated in its importance for the general organism, may pose some problems before heart surgery and antibiotic therapy previously may seem necessary. Finally, therapeutic consequences from emergencies in the dental office are reviewed.


A review of 700 bone marrow aspirations in Papua New Guinea. A retrospective survey of 700 marrow examinations in Papua New Guinea in a 2 1/2 year period has been carried out. Over half of the total came from the Port Moresby General Hospital. Anaemia and splenomegaly were the commonest indications. A disturbing proportion of the specimens were unsatisfactory for interpretation. Hypercellular marrows with erythroid hyperplasia were very common. Iron stores were absent or nearly absent in almost half of the evaluable specimens; this proportion rose to over 95% amongst pregnant or post-partum women with anaemia. Other relatively frequent findings included: a combination of features consistent with hypersplenism; mild degrees of eosinophilia and/or plasmacytosis; and mild degrees of megaloblastic changes. Haematological malignancies were found in over 10% of evaluable specimens. Of 611 assessable marrows from Melanesians, only 11 (1.8%) were entirely normal, but another 151 (24.7%) contained only minor abnormalities.


Potentially dangerous group O blood: incidence of anti-A and anti-B haemolysins in group O blood in Port Moresby. Samples of serum from 322 group O donors taken at random at Port Moresby, were examined within 24 hours of collection of blood by a simple screening test for haemolysins; 101 (35.31%) of 286 Papua New Guinean (PNG) group O sera and 16 (44.44%) of 36 European sera containing either anti-A or anti-B haemolysins. There were no differences in the extent to which A or B cells from Europeans or Papua New Guineans were lysed by these sera.


Hereditary ovalocytosis in Melanesians.

Hereditary ovalocytosis in Melanesians. A distinctive type of hereditary ovalocytosis has been found in Papua New Guinea and a few areas of Southeast Asia. Its main features include a high incidence among tropical lowland dwellers, autosomal recessive inheritance, specific depression of a number of red cell antigens, a characteristic morphology in blood films, and an effect on the erythrocyte sedimentation rate. Speculation has occurred as to whether the high incidence of ovalocytosis in malarious areas may be related to a selective advantage possessed by ovalocytics with regard to severe malaria. Preliminary data tend to support this hypothesis, but the evidence is not conclusive and much further work is needed.


Splenectomy: follow-up of patients in the Solomon Islands. The case histories of 19 splenectomy patients are discussed. Of these, four have died, five have had subsequent attacks of malaria, and the remainder have enjoyed good health, although only two patients have been taking regular malaria prophylaxis in this malaria endemic area. The roles of the reticulo-endothelial system and the spleen are discussed in relation to the effects of splenectomy.


Fiberoptic bronchoscopy in the diagnosis of lung cancer comparison of pre-and post-bronchoscopy sputa, washings, bruchings and biopsies. Fiberoptic bronchoscopy (brushings, washings and biopsies) was performed and pre- and post-bronchoscopy sputum cytologies obtained on 70 patients with histopathologically proven lung cancer. Bronchoscopy, with its associated procedures performed in 52 patients with primary bronchogenic carcinoma, was diagnostic in 41 (79%). Of all the various methods of obtaining specimens, bronchial brushing and bronchial biopsy gave the highest percentage yield (67%). However, since brush specimens could be obtained from peripheral lesions under fluoroscopic guidance, a greater number of positive specimens were obtained by this procedure (34) than by forceps biopsy (25), making brushing more useful. Pre- and post-bronchoscopy sputa were positive and thus of value in two cases when brushing and biopsy were both negative. Bronchial washing did not add significantly to the yield of positives and could therefore be eliminated as an unnecessary cost-and time-consuming procedure. Fiberoptic bronchoscopy was not helpful in diagnosing mediastinal tumors (5), lung metastases (7) and bronchial adenomas (6).


The morphology of cells in effusions settled on glass. The cells of human body cavity fluids were examined by light microscopy and with a scanning electron microscope after settling on the surface of glass. Some differences in behavior of various cell types were noticed. The histiocytes, a few minutes after settling, began to spread, and thin processes later extended from their cytoplasm. Extension of mesothelial cells was observed a few hours after settling, and was not proceeded by flattening. The flattening of neutrophils occurred soon after settling, and was not followed by extension. The other cells encountered in effusion, both benign and malignant, did not change their shape even up to 12 hours after the time of settling. The differences in behavior after the settling of various cell types encountered in human effusions is discussed in relation to their biological properties.


The accuracy and consistency of the cytologic classification of squamous lesions of the uterine cervix. The slides of 60 cases of cervical lesions in the five categories of benign inflammatory change, mild to moderate dysplasia, severe dysplasia, squamous carcinoma in situ, and invasive squamous carcinoma were reviewed on two separate occasions by two cytopathologists, nine cytotechnologists, and four cytotechnology students. Their overall concurrence of the cytologic with the tissue diagnoses was 78 per cent and was slightly higher for the cytotechnology students than for the more experienced observers. In 44 per cent of the invasive carcinomas and in 38 per cent of carcinomas in situ, there was considerable disagreement between cytologic and tissue classification partly due to the presence of inadequate material on the cytologic slides which had been selected consecutively based on the subsequent tissue diagnosis. The pathologists reproduced their own cytologic classification within one category variance in 94 per cent of the cases as compared to 92 per cent for the cytotechnologist and 86 per cent for the students. The study emphasizes the need for evaluation of laboratory performance based on routine diagnostic material in considering the accuracy and reproducibility of the procedure and in establishing quality control programs.


Primary papillary carcinoma of the prostate: report of a histopathologic, cytologic and electron microscopic study on one case. An unusual type of prostatic carcinoma is described. Histopathologic examination showed a papillary and acinar appearance. Aspiration cytology showed papillary fragments and sheets of malignant cells. The ultrastructural features of this tumor were similar to those reported for prostatic carcinoma rather than endometrial carcinoma supporting a prostatic orgin for this tumor.


Differential diagnosis between follicular adenoma and follicular carcinoma of the thyroid by nuclear DNA determination. Distinction between follicular adenomas and follicular carcinomas of the thyroid using fine needle aspiration cytology presents a problem. We investigated whether Feulgen photometric nucear DNA determination could offer additional, usable information for differential diagnosis. Since both diploid carcinomas and polyploid, widely scattered adenomas and nodular colloid goiter are seen, we found that nuclear DNA determinations proved useful in differential diagnosis only for occasional special cases.


Sarcoma of the vagina an unusual cytologic diagnosis. The reported case, exceptional in our oncological material, illustrates the cytologica features of sarcoma of the vagina. We believe that the diagnosis of sarcoma of the cervix or other parts of the female genital system is not difficult providing that the tumor is exfoliating cells. Sarcomatous cells should not be mistaken either with those from keratininzing epitheliomas with desquamated fiber cells or snake cells. Such differentiation should not be a problem for an experienced cytologst. In spite of the low frequency of this malignancy, the microscopical appearance of the cells is so different from the cells of routine gynecological material that when present we should, at least, suspect a sarcoma.


Microinterferometric measurement of dry weight of blood lymphocytes in hematologically normal patients and in patients with chronic lymphocytic leukemia. Lymphocytes from the peripheral blood of 20 patients with nonhematologic disorders (HN patients) and 13 patients with chronic lymphocytic leukemia (CLL) were studied. A scanning and integrating microinterferometer was used for calculation of dry weight of cells. In HN patients dry weight of lymphocytes was found to be 38.09 +/- 11.11 pg. The HN patients could be divided into three groups according to their lymphocytes' mund to be 29.79 +/- 7.99 pg. This population was uniform and resembled the "light" group of lymphocytes in HN patients. A study of the kinetics of PHA transformation of lymphocytes from both HN and CLL patients revealed a good correlation between the morphological evaluation and dry weight calculations.


Mechanisms of chromatid breakage in human lymphocyte cultures. G2 banding of human peripheral blood cultures with actinomycin D and tetracycline produced chromatid breakage in lightly stained Giemsa bands, and the number of such breaks tended to increase with fixation time. Chromatid breakage due to 3H-thymidine incorporation into DNA was also localized in light bands, but the number of these breaks did not increase with fixation time. These findings suggest that modification of chromosomal protein as a result of exposure to AMD or other chemicals following DNA synthesis can result in fixative-dependent chromatid breakage of susceptible chromosomal regions, unlike breakage due to 3H-thymidine which primarily affects DNA and is not affected by fixation. Thus, chromatid breakage observed in short-term lymphocyte cultures is not necessarily evidence of mutagenicity involving DNA, but rather may be due to toxic effects on synthesis or attachment of chromosomal proteins.


Cytogenetic studies of human endometrial carcinomas by means of tissue culture. The cytogenetic studies of human endometrial carcinomas were performed using a tissue culture technique. Seventeen cases or 40.5 per cent out of 42 trials for culture could be analyzed. The mode of chromosome number was found at the diploid range in all cases, and the exact counting of the modal range revealed that the peak of the number was 46 in 75 per cent of the analyzed cases. The distributions were even both in the hypo- and hyperdiploid areas. The karyotype analyses showed that the majority of the endometrial carcinomas presented pseudo-diploid chromosomes, revealing irregular numerical alterations of each group constitution and scattered variant chromosomes with minor structural changes. Only four cases were noted to have an unclassified chromosome with marked structural aberrations. A marker chromosome was identified in only one case, whereas no common specific chromosome was noted and the histologic findings did not correlate with chromosome aberration. From our tissue culture study, carcinomas of human corpus uteri consist mainly of the cells with a pseudodiploid chromosomal constitution. It is suggested that a culture method would be a desirable procedure for the cytogenetic study.


Psammoma bodies of benign endometrial origin in cervicovaginal cytology. This case report illustrates a benign endometrial condition associated with a combination oral contraceptive agent in which psammoma bodies were produced. Additionally, it reemphasizes the fact that nonmalignant processes are capable of forming such structures.


A quick method for concentrating and processing cancer cells from serous fluids and fine-needle nodule aspirates. A quick method of concentrating cancer cells and the preparation of cytological slides from body fluids and aspirates is described, using a single Ficoll gradient and cytocentrifuge. The method eliminates the disadvantages of conventional techniques, i.e., excess contamination by red (erythrocytes) or white (leukocytes) blood cells and a scarcity of cancer cells. Because the technique is simple and requires only standard cytotechnological equipment, it can be easily adopted as an aid to diagnostic routine in cancer cytology.


McArdle's syndrome. Fine structural changes in muscle. Two cases of McArdle's syndrome are reported. One is a "classical" example; the other is unusual because of the in vitro presence of muscle phosphorylase activity. In the latter case, the electronmicroscopic investigation confirmed the diagnosis. The fine structural changes characteristic of this disease are summarized and it is concluded that histochemical studies alone are insufficient to exclude the diagnosis of McArdl's myopathy.


Pharmacokinetic and pharmacodynamic studies on oestradiol valerianate administered orally to postmenopausal women. Peripheral plasma from four postmenopausal women was analysed for oestrone (E1) and oestradiol (E2) during 48 hours after oral intake of a single tablet of 2 mg Progynon (oestradiol valerianate). Plasma levels of E1, E2 and d-norgestrel were analysed daily in five postmenopausal women during treatment with tablet Cyclabil (oestradiol valerianate in a biphasic preparation with dl-norgestrel) in 21 dyas. Radioimmunoassay (RIA) was utilized for the quantifications. Pretreatment plasma levels of E1 were about 20 pg/ml and E2 about 12 pg/ml. It is concluded that oestradiol valerianate is rapidly absorbed from the gastrointestinal tract and converted to E1. This is reflected by plasma levels of E1 considerably higher than those of E2. The E2 levels found were in the range of those in ovulatory women, while the oestrone levels were higher.


The prevention and management of post-menopausal osteoporosis. In 6 groups of peri- and post-menopausal women, there was an inverse relation between the urinary sediment smear maturation value and the fasting urinary hydroxyproline/creatinine ratio. Administration of ethinyloestradiol and Progynova both reduced urinary hydroxyproline into the pre-menopausal range, the fall being proportional to the starting value. Oestrogen therapy also produced a significant fall in plasma ionised calcium. In a prospective trial, oestrogen therapy prevented post-menopausal bone loss but calcium therapy was less effective. It is suggested that a high fasting urinary hydroxyproline/creatinine ratio might be taken as an indication for oestrogen therapy in post-menopausal women. In established post-menopausal osteoporosis, pre-disposing risk factors appear to be low calcium intake, malabsorption of calcium and low oestrogen status. These patients appear to represent the fast bone-losers in the post-menopausal population. The accelerated bone loss can be wholly or partially corrected by hormone replacement therapy and by calcium supplements given to those with normal absorption only. These therapies also prevent loss of height due to further crush fractures. The malabsorption of calcium is very resistant to vitamin D therapy but responds to 1alpha-OHD3. Balance data suggest that the most effective therapy may be a combination of 1alpha-OHD3 with oestrogen.


The perception of menopause in five ethnic groups in Israel. An investigation was made into the menopausal symptoms and attitudes of 1, 148 Israeli women of five different ethnic origins. The results show that, while some of the somatic menopausal symptoms are independent of ethnic origin, psychic and psychosomatic symptoms are more closely associated with this variable. Similarly, attitudes to the climacteric vary across cultural origins, especially with regard to husband-wife relationships. This study underlines the necessity of a multi-factorial, biological, sociological, psychological and anthropological approach to problems of menopause.


Pregnancy in the young teenage woman.

Pregnancy in the young teenage woman. Since 1967 the number of delivered young teenage women has gradually decreased. During the same period the number of therapeutic abortions among these young women has increased, probably to a sufficient extent to explain the decreased nativity. There is no evidence in this study of any effect of more extended sex and contraceptive education in schools.


The effect of oxytocin on hypertonic saline abortion. Oxytocin was infused in 22 randomly selected pregnant women after extraamniotic hypertonic saline instillation. In another 24 pregnant women no oxytocin was infused at midtrimester abortion. There was no difference between the two groups in mean time from saline injection to expulsion of the fetus.


Dysgerminomas (seminomas) in genetic males with female phenotype. One case of gonadal dysgenesis and gonadoblastoma and one of testicular feminization. Gonadal tumors in patients with somatosexual ambiguity are rare but of considerable clinical and pathological interest. Two patients with dysgerminomas and abnormal sexual differentiation were recently treated at the Department of Gynecologic Oncology. One (Case 1) was an 18-year old girl and the other (Case 2) an unmarried woman of 58 years. In spite of their female phenotype both showed a male genotype, 46, XY. In Case 1 the dysgerminoma (or rather seminoma) originated in a gonadoblastoma in one of two dysgenetic testes but in Case 2 in one of two "feminizing testes". The clinico-pathological features of both cases will be reported and commented and the relation between the germ cell tumors, gonadal dysgenesis and the Y-chromosome will be discussed.


The influence of the bony pelvis in persistent occiput posterior position. Roentgenologic pelvimetry has been performed in 68 consecutive cases of occiput posterior position. The results point to an influence from a transverse narrowing of the pelvis or a tendency towards large dimensions. In many cases, however, no clear correlation to pelvic dimensions is found. A significant difference exists in the rate of obstetrical complications between nulliparous and parous women. The reasons for this and for the persistent OP position are discussed.


Transcutaneous monitoring of PO2 in different skin areas in the neonate and in the scalp of the fetus during labor - methodological and physiological observations. The reliability and physiological significance of continuous non-invasive PO2 recording by means of skin electrodes was studied by comparing simultaneous tracings from different skin areas in a series of 40 newborns with normal labor and birth. A new measuring equipment - the Radiometer prototype unit TPM 1 - was used. In addition, the applicability of the method to the fetus was tested in a primary series of 11 labor cases. Certain technical and physiological factors that can influence the tracings were identified.


Efficacy of primary radiation in carcinoma of the endometrium. Five-year follow up of unselected material from a specific region in the north of Sweden. A case material from the northern part of Sweden comprising all known cases of carcinoma of the endometrium during the years 1963-1969 is presented. A clinical experiment has been carried out in which 320 patients with carcinoma of the endometrium have been uniformly treated. The patients have been given primary radiation treatment, mostly with radium packing according to a modified Heyman method. About two thirds of the patients have had a prophylactic operation with total removal of the uterus and the adnexa about six weeks after the radiation treatment. Patients with contraindications to surgery have had a curettage three months after radiation treatment. The primary cure rate and the complications have been reported in a previous paper. The five-year survival rate in stage I in cases treated with radiation and surgery was 92%. The survival rate corrected for intercurrent death was 93%. The survival rate in cases treated with radiation only was 71%. The survival rate corrected for intercurrent death was 88%. The total survival rate in stage I was 85% and the survival rate corrected for intercurrent death in Stage 2 was 92%. The total survival rate in the whole case material including all stages was 75.3% and the survival rate corrected for intercurrent death in the total material was 82.8%.