Neuronal activity in the lateral vestibular necleus of the cat. IV. Postsynaptic potentials evoked by stimulation of peripheral somatic nerves. The synaptic input to Deiters neurones evoked by stimulation of peripheral somatic nerves was measured by intracellular recordings. EPSPs with broad receptive fields and latencies which indicate polysynaptic connexions were commonly evoked from the FRA. In other cells, low threshold cutaneous afferents were effective at rather short latencies suggesting oligosynaptic connexions from fast ascending fibres. One example was found of EPSPs due to low threshold muscle afferents. IPSPs due to climging fibre activation of Purkinje cells as observed in most of the neurones were evoked by cutaneous volleys above 1.5-2.0T and muscle volleys above 5T (above 3-3.5T in case of Q). Often, IPSPs were evoked by stimulation of nerves, to the segmental level of which the the vestibulospinal neurone under investigation projected. A small proportion of cells received short latency IPSPs involving direct fast mossy fibre tracts, which were evoked from low threshold cutaneous afferents. IPSPs due to polysynaptic mossy fibre activation of Purkinje cells were evoked from the FRA bilaterally and from ipsilateral cutaneous afferents at 1.5-2.0T ("prolonged inhibition"). Prolonged excitatory/inhibitory events mediated by mossy fibre pathways may be involved in quadruped locomotion or other processes making use of a broad motor integration.


Changes in chemosensitivity of developing chick muscle fibres in relation to endplate formation. 1. The sensitivity to acetylcholine (ACh) of the fast posterior latissimus dorsi (PLD) and slow anterior latissimus dorsi (ALD) during embryonic development was studied and compared. The sensitivities were expressed as a ratio of the maximal tetanic tension and tension developed in response to ACh. 2. Up to the 17th day of incubation both muscles are sensitive to ACh to a similar extent; at the 18th day the sensitivity of the PLD muscle decreases and continues to do so until hatching and thereafter. 3. Since the decrease in sensitivity of PLD muscles takes place a few days after innervation, it is suggested that this is caused by activity of the motor nerve. To test this curare (dTc) and hemicholinium (HC-3), drugs that interfere with neuromuscular transmission, were injected into the yolk sac of the embryos when nervemuscle connections are usually established. In the curare and HC-3 treated embryos the desensitization of the PLD muscles did not take place. 4. The distribution of endplates on PLD muscles from drug treated 20-21 day old embryos was compared to that of untreated controls. Whereas control PLD muscles have only one band of endplates, muscles from curarized embryos and HC-3 treated embryos have several bands of endplates, and many muscle fibres with multiple innervation were found. 5. It is suggested that nerve fibres which make connections with PLD muscle fibres bring about a decline in chemosensitivity by releasing more transmitter, and thereby prevent further nerve muscle connections from being made along the same muscle fibre.


[Scintigraphy of the choroid plexus (choroid plexography)]. An increase concentration of pertechnate in the choroid plexus was demonstrated in 21 patients pretreated with tin pyrophosphate injected intravenously 24 hours before the examination. Using this technique, the choroid plexus are seen quite clearly, giving indirectly a picture of the intracerebral ventricles. A dynamic study was carried out in 12 patients, 3 of whom had abnormalities of the cerebrospinal fluid circulation demonstrated by isotope cisternography. The results obtained were characterised by a typical diphasic curve with variations in the pathological cases.


[Ultrastructural abnormalities of the chondrocytes in pycnodysostosis. Their relation to a disorder of lipid metabolism]. The ultrastructural study of the growth cartilage of pycnodysostosis reveals the presence of abnormal inclusions in the majority of the chondrocytes. The inclusions are single membrane bound and contain granular material and lamellar irregularly interwoven structures which at very high magnification appear to be made up of dense parallel bands. These vacuoles displace adjacent structures and some of them appear to be closely related to the Golgi apparatus. In addition, appearances are sometimes seen which suggest the expulsion of the vacuoles into the cell capsules. The abnormal chondrocyte inclusions are visible by optic microscopy and stained with Nile blue on frozen section. Thus, histochemical and ultrastructural characteristics suggest a lipid (probably phospholipid) content. Many authors have already stressed the role of lipids in the process of calcification. The abnormalities described might bear some relation to the densification of the skeleton seen in pycnodysostosis.


Prazosin: biochemistry and structure-activity studies. Prazosin, a structurally and mechanistically novel antihypertensive agent, is the culmination of a systematic synthesis and pharmacologic developmental effort which had as its goal the development of an agent that would lower blood pressure by direct vasodilation. Pharmacologic studies have demonstrated that the vasodilator action of prazosin is confined primarily to the arterioles and that antihypertensive effects are unaccompanied by excessive increases in heart rate. Evidence is presented rationalizing these observations on a biochemical basis. Results of inhibitory studies with isolated cyclic nucleotide phosphodiesterases are consistent with the hypothesis that prazosin elicits its therapeutic effects by increasing intracellular levels of cyclic AMP at vascular sites and intracellular levels of cyclic GMP at cholinergic receptor sites in the heart. Tissue distribution studies with radioactive prazosin-2-14C are in accord with this rationale and further support the mechanistic conclusions reached on the basis of pharmacologic observations. Pharmacokinetic and biotransformation studies indicate that prazosin is well absorbed and is excreted principally in metabolized form with biliary excretion being the major route of elimination. A high margin of safety for prazosin has been established in safety evaluation studies in a variety of animal species.


Renal transplantation in the very young child. Thirteen children, aged between one and five years, received transplants and 11, all related-donor recipients, are alive (mean follow-up 2.2 yr). Five had congenital nephrotic syndrome, 5 had hypoplastic-dysplastic kidneys and one each had polycystic kidneys, nephroblastoma and glomerulonephritis. One child died of peritonitis and one died following irrevocable rejection. Two children required retransplantation. The present serum creatinine of the survivors is 0.67 +/- 0.23 (x +/- SD)mg/100ml. Eight of the 11 survivors have shown catch-up growth. Thus we conclude that related-donor transplantation is warranted in children having reached the age of one year and six kilograms in body weight.


Circadian rhythm of plasma renin activity and plasma aldosterone in normal man and in renal allograft recipients. In normal supine man, a marked circadian rhythm was found in plasma renin activity (PRA) and in plasma aldosterone concentration, with highest levels in the morning and lowest in the evening. Short-term suppression of ACTH by dexamethasone did not eliminate the circadian variation of plasma aldosterone. In patients with a transplanted and thus denervated kidney, no such rhythm of PRA could be observed. From our data it is suggested that the normal circadian rhythm of PRA depends on intact renal innervation and is controlled by the central nervous system.


Pulmonary infections after kidney transplantation. Seventy-five pulmonary infections out of 173 kidney transplantations have been observed in 62 graft-recipients, from 1965 to 1973. The aetiologic organism is a bacteria in 80%  of the cases, a fungus in 8% of the cases, cytomegalovirus and Pneumocystis Carinii in 11% of the cases. A close relationship between infection and a previous transplant crisis was noted in 66% of the cases. The most efficient diagnostic procedures were pleural and open lung biopsy as well as cultures of blood, pleural fluid and tracheobronchial secretions. Treatment was effective in 60% of bacterial and Pneumocystitis Carinii infections; on the other hand, it was less useful in fungal infections and totally ineffective in cytomegalovirus infections. Despite a possible increase in the risk of rejection, reduction or even arrest of immunosuppressive therapy is recommended in severe infections.


Control of oscillating glycolysis of yeast by stochastic, periodic, and steady source of substrate: a model and experimental study. Type and range of entrainment of glycolytic oscillations by a periodic source of substrate are determined experimentally in yeast extracts. Subharmonic entrainment proves the nonlinear nature of the glycolytic oscillator Random variation of the substrate input yields sustained oscillations of irregular waveform and stable period. The results agree with the predictions of an allosteric model for phosphofructokinase (EC 2.7.1.11; ATP:D-fructose-6-phosphate 1-phosphotransferase), which is the enzyme responsible for periodic operation of glycolysis. A comparison between model and experiment in the case of a constant source of substrate further indicates that the oscillatory dynamics of the glycolytic system can satisfactorily be described by the phosphofructokinase model.


Partial purification of a glucocorticoid receptor. A simple method for purification of the glucocorticoid receptor from hepatoma tissue culture cells has been developed. The procedure, which requires only about 24 hr, involves biospecific adsorption of the receptor to deoxycorticosterone derivatized agarose, elution with a glucocorticoid, and gel filtration. The receptor-steroid complex is obtained in 35-40% yield and is about 2000-fold purified. It possesses properties similar to those reported in crude extracts, including sedimentation coefficient in glycerol gradients and activation-dependent binding to nuclei.


Oxygen and cyclic nucleotides in human umbilical artery. In the human umbilical artery O2 has a direct contractile effect and is also required for induction of contraction by several other agents. Agonist that cause contraction (bradykinin, histamine, and serotonin) cause accumulation of guanosine 3':5'-monophosphate (cGMP) without altering adenosine 3':5'-monophosphate (cAMP). They appear to act through two different mechanisms: one Ca++-dependent, the other Ca++-inhibited. O2 increased the cGMP content of the artery in a Ca++-dependent manner without affecting the cAMP content. Inhibitors of oxidative phosphorylation (oligomycin and 2,4-dinitrophenol) did not diminish this effect of O2. O2 was required for demonstration of the Ca++-dependent accumulation of cGMP in response to bradykinin, histamine, and ionophore A23187. The effect of the phosphodiesterase inhibitor 3-isobutyl-1-methyl xanthine on basal cGMP content and on the bradykinin-induced accumulation was also dependent on the presence of O2. Methylene blue and sodium ascorbate caused cGMP accumulation in O2-deprived arteries. Their effects were not diminished in Ca++-depleted arteries and, in fact, seemed to be inhibited when 2.7 mM Ca++ was present in the medium. The effects of these agents and of serotonin on cGMP, which were inhibited by Ca++, were also inhibited by O2. These non Ca++-, non O2-dependent agents (methylene blue, ascorbate, and serotonin) did not, however, permit demonstration of the effects of the Ca++- and O2-dependent agonists on O2-deprived arteries. It appears that there are in the umbilical artery (and probably in other tissues also) at least two separate mechanisms for control of cGMP synthesis that are influenced differently by Ca++- and O2-linked processes.


Dynamic and static quenching of 1,N6-ethenoadenine fluorescence in nicotinamide 1,N6-ethenoadenine dinucleotide and in 1,N6-etheno-9-(3-(indol-3-yl) propyl) adenine. For nicotinamide 1,N6-ethenoadenine dinucleotide (epsilonNAD+), the fluorescent analog of NAD+, in neutral aqueous solution the quantum yield has been determined to be 0.028 and the fluorescent lifetime, 2.1 nsec. Simultaneous determination of quantum yields and lifetimes of epsilonNAD+ and of the "half molecule" epsilonAMP allows the calculation of the percentage of stacked and open conformations of the dinucleotide. At 25 degrees in neutral aqueous solution there is 45 +/- 5% of stacked forms. The value of the fluorescent impurities, especially those containing the epsilon-adenosine moiety, and a purification procedure using high performance liquid chromatography was devised to obtain fluorescently homogeneous preparations. In order to study the effect on epsilon-adenosine fluorescence caused by the possible close proximity of a tryptophan in a polypeptide chain or protein, we have prepared 1,N6-etheno-9-[3-(indol-3-yl)propyl]adenine (epsilonAde9-C3-Ind3), a model compound in which indole is used as a neutral substitute for tryptophan. Fluorescence studies on epsilonAde9-C3-Ind3 show that the formation of an intramolecular complex results in complete quenching of the epsilon-adenine fluorescence. It is therefore predictable that positioning of the epsilon-adenosine of any fluorescent coenzyme moiety (e.q., epsilonATP, epsilonADP) in close proximity to a tryptophan in a protein will result in complete fluorescence quenching of the former.


Biosynthesis of mouse interferon by translation of its messenger RNA in a cell-free system. A fraction of mouse RNA containing messenger RNA coding for mouse interferon was translated with high efficiency in a wheat germ system into a fully active product. This product fulfills the criteria for mouse interferon, namely: (1) it was active against vesicular stomatitis virus and herpes simplex virus in mouse cells; (2) its antiviral activity was species specific; (3) its activity was completely neutralized by mouse anti-interferon serum. The synthesis of interferon in this cell-free system requires the presence of spermine.


Specific endocrine and hormonal receptor changes associated with liver regeneration in adult rats. Partial removal of the adult rat liver--a procedure that stimulates liver regeneration--causes arterial levels of insulin and thyroxin to fall and glucagon levels to rise. These changes are correlated with the quantity of liver tissue excised and not solely with nonspecific operative manipulations or with blood glucose levels. During regeneration (about 24 hr postoperatively), hepatic plasma membranes become partially resistant to binding glucagon; this change is specific because it is not detected in laparotomized controls and insulin binding is not significantly altered. It remains to be proven whether any one or more of these specific endocrine changes are casually related to the initiation of hepatic proliferation.


Effect of oncogenic virus on muscle differentiation. Chick muscle cultures infected with wild-type Rous sarcoma virus form myotubes, but these myotubes vacuolate and by day 6 most have degenerated, leaving only large numbers of transformed mononucleated, replicating cells. Muscle cultures infected with a temperature-sensitive mutant (TS) at permissive temperatures behave as cells infected with wild-type Rous sarcoma virus. TS-infected cells reared for 8 days at nonpermissive temperature form contracting myotubes, plus large numbers of fibroblastic cells. If these cultures are lowered to permissive temperature, within 72 hr the myotubes vacuolate and degenerate, whereas the mononucleated cells transform. If replicating TS-transformed cells after 8 days at permissive temperature are shifted to nonpermissive temperature, within 72 hr many cells fuse and form contracting, post-mitotic myotubes. Creatine kinase (ATP:creatine N-phosphotransferase, EC 2.7.3.2) levels parallel the formation and degeneration of myotubes during these temperature shifts. If the viral transforming gene is expressed in the post-mitotic myotubes it is lethal, whereas it is not lethal if expressed in replicating percursor myogenic cells. The viral gene expression at permissive temperature blocks further myogenesis depending on the position of the cells in the myogenic program. The virus does not cancel the replicating, transformed myogenic cells' commitment to, or position in, the myogenic lineage. When the transforming action of the virus is suppressed, the normal myogenic program resumes.


Restoration of ultraviolet-induced unscheduled DNA synthesis of xeroderma pigmentosum cells by the concomitant treatment with bacteriophage T4 endonuclease V and HVJ (Sendai virus). Ultraviolet (UV)-induced unscheduled DNA synthesis of xeroderma pigmentosum cells, belonging to complementation groups A, B, C, D, and E, was restored to the normal level by concomitant treatment of the cells with T4 endonuclease V and UV-inactivated HVJ (Sendai virus). The present results suggest that (1) T4 endonuclease molecules were inserted effectively into the cells by the interaction of HVJ with the cell membranes, (2) the enzyme was functional on human chromosomal DNA which had been damaged by UV irradiation in the viable cells, (3) all the studied groups of xeroderma pigmentosum ("variant" was not tested) were defective in the first step (incision) of excision repair.


Possible peptide chain termination mutants in thymide kinase gene of a mammalian virus, herpes simplex virus. Mutations in the viral gene coding for the thymidine kinase (ATP:thymidine 5'-phosphotransferase, EC 2.7.1.75) induced by herpes simplex virus have been obtained by selection of virus resistant to bromodeoxyuridine when grown in thymidine-kinase-deficient LMTK- mouse cells. Proteins labeled after infection of Vero (monkey) cells with herpes simplex virus were analyzed by gel electrophoresis and one protein of about 40,000 daltons was consistently altered in a number of thymidine-kinase-deficient mutants. Many viral mutants lacked this peptide and one class of these mutants induced the synthesis of new shorter peptides. Revertant virus could be selected which simultaneously regained the ability to induce thymidine kinase activity, regained the intact thymidine kinase peptide, and lost the ability to synthesize the shorter peptide fragment. These mutants comprise a class of animal virus mutants which have the properties expected of peptide chain termination mutants.


Evolution of type C viral genes: preservation of ancestral murine type C viral sequences in pig cellular DNA. Domestic pigs (Sus scrofa) and other members of the family Suidae have multiple copies of type C viral gene sequences in the cellular DNA of all their tissues. Partially homologous viral gene sequences are also found in cellular DNA of rodents, particularly Muridae. The results lead to the conclusion that type C viral genes were introduced into the Suidae lineage as a result of trans-species infection by an ancestral xenotropic murine virus. The rate of evolution of the virogene sequences in the pig appears to be much slower than that of genes that have remained in the rodent lineage; this may be a consequence of transfer from a shorter-lived animal (the rodent) to a longer-lived one (the pig). We estimate the time of gene transmission as 5-10 million years ago and conclude that the present-day porcine type C virogenes most closely approximate the viral genes as they were several million years ago in the rodent lineage.


A unique complement derived chemotactic factor for tumor cells. A factor chemotactic for Walker carcinosarcoma and Novikoff hepatoma tumor cells can be generated by incubation of serum with an extract from tumor cells. The chemotactic factor has no activity for neutrophilic leukocytes, and three factors chemotactic for leukocytes are not chemotactic for tumor cells. By the use of complement deficient serums as well as purified complement components, the chemotactic factor for tumor cells has been found to be a fragment of the fifth component (C5) of complement and appears to result from direct interaction of C5 with an enzyme in the extract. The chemotactic factor for tumor cells can be classified as a functionally unique fragment of C5 that may significantly influence the behavior of tumor cells in vivo.


Growth and differentiation in culture of leukemic leukocytes from a patient with acute myelogenous leukemia and re-identification of type-C virus. Conditioned medium from a culture of whole human embryo cells stimulated prolonged exponential growth in suspension culture of leukocytes from a patient with acute myelogenous leukemia. Ten to 20% of the cultured cells consistently differentiated into mature granulocytes including neutrophils, eosinophils, and basophils. The proportion of lymphocytes declined after culturing, and tests for Epstein-Barr virus antigens were negative. An abnormality of a G group chromosome was observed in some metaphases from the patient's fresh bone marrow and from the cultured leukocytes, indicating growth in vitro of leukemic cells. After 4-10 weeks in culture, a budding type-C virus was continuously released by the cultured leukocytes, predominantly by undifferentiated blast cells. This virus was originally identified in three different cultures of a peripheral blood specimen obtained at the time of diagnosis. Subsequently, this virus was identified by reverse transcriptase (RNA-dependent DNA polymerase) assays and by electron microscopy in cultured leukocytes from a bone marrow specimen obtained 14 months later from the same patient. Virus produced by cultures of both specimens was closely related, if not identical, to the woolly monkey type-C virus.


Complete translation of poliovirus RNA in a eukaryotic cell-free system. Poliovirus RNA stimulates imcorporation of 35S from both [35S]methionine and formyl-[35S]methionyl-tRNAfMet in cell-free systems derived from HeLa cells or from poliovirus-infected HeLa cells. The largest product formed under the direction of the viral RNA is the same size as the polyprotein thought to represent translation of the entire RNA. Synthesis of this polyprotein and other large products was stimulated greatly by increasing the salt concentration during the reaction from the optimum for initiation (90 mM) to the optimum for elongation (155 mM). Only one initiation peptide could be identified, and a tryptic digest of the product contained mainly peptides that cochromatographed with peptides from authentic viral proteins. The RNA from a deletion mutant of poliovirus initiated protein synthesis at the same site used by standard RNA and programmed synthesis of an appropriately deleted set of polypeptides. The results strongly support the model of translation of poliovirus RNA from a single initiation site into a continuous polyprotein that is cleaved to form the functional proteins. It is suggested that uninfected HeLa cell extracts can carry out the cleavages of nascent polyprotein.


Effects of estrogen on gene expression in chick oviduct: nuclear receptor levels and initiation of transcription. Estrogen (diethylstilbesterol) was administered in vivo to chicks for various time periods. Chromatin was then prepared from oviduct nuclei and assayed for its capacity to support initiation of RNA chain synthesis in vitro in the presence of saturating levels of Escherichia coli RNA polymerase (RNA nucleotidyltransferase; nucleosidetriphosphate:RNA nucleotidyltransferase; EC 2.7.7.6). These same nuclei were also assayed by a [3H]estradiol exchange assay for their endogenous receptor content. The number of available initiation sites for RNA synthesis on chromatin was shown to correlate with the endogenous levels of nuclear estrogen receptor. A decrease in the nuclear concentration of estrogen receptor molecules and the concentration of initiation sites for RNA synthesis occurred during withdrawal of estrogen from previously stimulated chicks. Both parameters declined with a similar half-life. When estrogen was readministered to withdrawn chicks, the number of initiation sites increased 2-fold as early as 30 min and approached a maximal level (3-fold) by 1 hr. During the same period of restimulation with estrogen, the number of estrogen receptor molecules bound to nuclei increased to a maximum at 20 min and then declined at 1 hr to a steady-state level 2-fold higher than the withdrawn chicks. Simultaneous measurements of RNA chain length and RNA chain propagation rate demonstrated that parameters remained relatively constant throughout estrogen withdrawal as well as secondary stimulation. The temporal correlation between changes in the levels of nuclear-bound estrogen receptor and the number of RNA chain initiation sites on chromatin prepared from these same nuclei strongly suggested that the hormone receptor complexes act on chromatin to mediate these changes in genetic transcriptional activity.


Anatomy of herpes simplex virus DNA: evidence for four populations of molecules that differ in the relative orientations of their long and short components. Intact DNA molecules extracted from HSV-1 (herpes simplex virus 1, human herpes virus 1) strain MP virions have a molecular weight of approximately 97 X 10(6), but cleavage with the HinIII restriction enzyme yields fourteen fragments with summed molecular weights of 160 X 10(6). Six "major" fragments occur once in every molecule in the population and account for 60% of the genetic information. Four "minor" fragments are present in amounts equivalent to one copy for every two genomes (0.5 molar ratio) and the other four occur only once in every four molecules (0.25 molar ratio). The minor fragments can be arranged into four equimolar sets, each with summed molecular weights that account for the remaining 40% of the genome. Treatment with lambda 5' exonuclease revealed that all molecules contain 0.5 molar ratio fragments at both termini. These observations and the results of similar analyses of the EcoRI and double HinIII/EcoRI digests indicate that there are four distinct structural forms of HSV DNA which differ only in the relative orientations of two subregions, designated L and S. The L and S segments consist of 82 and 18% of the sequences, respectively, and each has inverted terminally redundant regions that correspond to the internal duplications observed by electron microscopy. The DNA from other strains of HSV-1 and 2 also consists of equal proportions of all four possible permutations of the L and S segments. These unusual features of HSV DNA molecules have novel implications with regard to the genetic map and the mode of replication and evolution of herpes simples viruses.


Conformational fluctuations of DNA helix. Treatment of a homogeneous sample of circular DNA molecules that contain at least one single-strand scission per molecule with ligase yields a population of covalently closed circles heterogeneous in the topological winding number alpha. Under proper conditions, the product molecules with different values of alpha can all be resolved by gel electrophoresis. The distributions in alpha for several DNAs, covalently closed at different temperatures, have been determined and the results have been interpreted in terms of thermal fluctuations of the DNA helix.


Translation of RNA that contains polyadenylate from yeast mitochondria in an Escherichia coli ribosomal system. RNA that contains poly(A) [poly(A)-RNA] has been isolated from yeast mitochondria by poly(U) Sepharose-4B column chromatography. Pulse-labeled poly(A)-RNA shows 8-10 discrete peaks by acrylamide gel electrophoresis. The specific activity of mitochondrial poly(A)-RAN is six to eight times greater than that of mitochondrial rRNA after pulse labeling of protoplasts with [3H-]uridine. Ethidium bromide inhibits incorporation by over 90%. The total mitochondrial RNA preparation was contaminated with 5-15% cytoplasmic rRNA as determined by gel electrophoresis, but RNA exhaustion hybridization experiments indicated little or no cytoplasmic contamination of the mitochondrial poly(A)-RNA. The poly(A)-RNA stimulates [3H]leucine incorporation into protein in an E. coli cell-free system. A fraction of the labeled product is precipitated with antibody directed toward yeast cytochrome oxidase, but not with antibody directed toward bovine serum albumin. Sodium dodecyl sulfate gel electrophoresis of the immunoprecipitated material reveals labeled peptides having the mobility of the three larger cytochrome oxidase peptides, which are known to be translated by mitochondrial ribosomes.


Androgen- and estrogen-binding macromolecules in developing mouse brain: biochemical and genetic evidence. Androgen- and estrogen-binding macromolecules from the hypothalamus plus preoptic area of 3- to 4-week-old mice have been detected and partially characterized. These components bind the respective hormones with high affinity (saturating at 4-8 nM) and sediment with rates typical of presumed steroid receptors (4.0-4.5 S in 0.15 M NaCl, 5.0-7.5 S without salt). A 90-95% reduction in androgen binding found in the androgen-insensitivity mutant mouse, testicular feminization (Tfm), provides a genetic control for the specificity of binding. This reduced androgen binding with Tfm/Y mutants and blocking experiments with non-radioactive estradiol [estra-1,3,5(10)-triene-3,17beta-diol] and testosterone (17beta-hydroxy-4-androsten-3-one) indicate the existence of at least two binding components: one with high affinity only for estradiol, the other with affinity for both androgens and estrogen. Based on these properties, a receptor mechanism that detects relative concentrations of androgens and estrogens is proposed.


Requirement for cellular protein synthesis in reversal of ethidium-bormide-induced inhibition of cell transformation by murine sarcoma virus. Cultures of mouse Balb 3T3 fibroblasts exposed to a noncytotoxic dose of ethidium bromide for 16-18 hr are unable to produce foci after infection with murine sarcoma virus. Such cultures regain susceptibility to infection when incubated for 6-8 hr in drug-free growth medium. Pretreated but not untreated cultures exhibit sensitivity toward brief (6 hr) exposure to cycloheximide, chloramphenicol, and actinomycin D before infection. Pretreatment with cordy-cepin inhibits the ability of cultures to produce foci after infection. The recovery of ethidium-bromide-treated cultures requires the synthesis of cellular proteins which may have some important role in the establishment of RNA tumor virus infection.


Effect of extradiol-17beta on the synthesis of specific uterine nonhistone chromosomal proteins. The synthesis of specific nonhistone chromosomal proteins in the uterus of the ovariectomized rat was examined as a function of time after treatment with estradiol-17beta. Sequential stimulations in the rates of synthesis of at least five nonhistone chromosomal proteins having molecular weights of 96,000, 70,500, 29,400, 20,700, and 16,400, respectively, were observed. The rate of synthesis of the nonhistone chromosomal protein having a molecular weight of 70,500 was increased at 1 hr after hormone treatment. This was the first nonhistone chromosomal protein to be induced by estrogen, and its induction was blocked by pretreatment with actinomycin D. The data reported suggest but do not prove that this protein is the 4.5 S estrogen receptor, and that it is the induced nuclear acidic protein described earlier by Teng and Hamilton. The rates of synthesis of the nonhistone proteins with molecular weights of 96,000, 29,400, 20,700, and 16,400 were increased at 3,5,24, and 24 hr, respectively, after hormone treatment.


Thermolabile T (tumor) antigen from cells transformed by a temperature-sensitive mutant of simian virus 40. Partially purified tumor (T) antigen from a strain of Chinese hamster lung cells transformed by wild-type simian virus 40 (SV 40) and either of two temperature-sensitive SV 40 mutants has been studied as a DNA binding protein. The DNA binding activity present in the T-antigen-containing fractions is inhibited by purified hamster anti-T IgG but not by equivalent amounts of nonimmune hamster IgG. T from either wild-type- or tsC219-transformed cells is relatively stable during heating at 44 degrees compared to T prepared from tsA239-transformed cells. These results strongly suggest that T is a product of the SV 40 A gene.


In vitro synthesis of a constitutive enzyme of Escherichia coli, 6-phosphogluconate dehydrogenase. Enzymes of central intermediary metabolism are among the constitutive proteins of Escherichia coli. 6-Phosphogluconate dehydrogenase [6-phospho-D-gluconate: nicotinamide adenine dinucleotide phosphate 2-oxidoreductase (decarboxylating), EC 1.1.1.44-A1, an enzyme of the hexose monophosphate shunt, was synthesized in vitro in a coupled transcription-translation system directed by DNA from a specialized transducing phage carrying gnd, the structural gene. Enzyme synthesized in vitro was detected by radiochemical assay of its activity. Kinetic experiments using inhibitors of transcription and translation suggest that synthesis of the mRNA for the enzyme was initiated at the gnd promoter. Neither cyclic AMP nor cyclic GMP had any effect on the amount of enzyme synthesized in vitro.


Tumorigenicity of virus-transformed cells in nude mice is correlated specifically with anchorage independent growth in vitro. Clonal isolates of mouse 3T3 cells and primary rat embryo cells, recovered nonselectively after infection by simian virus 40 (SV40), have been tested for tumorigenicity in the immune-deficient nude mice in order to determine the cellular growth properties in vitro specifically correlated with neoplastic growth in vivo. In addition, mouse 3T3 cells transformed by murine sarcoma virus (MuSV, Kirsten strain), and revertants isolated from cells fully transformed by either SV40 or MuSV were also studied. Results suggest that the single cellular property consistently associated with tumorigenicity in nude mice is the acquisition by virus-transformed cells of the ability to proliferate in vitro in the absence of anchorage. Other cellular parameters of virus-induced transformation, such as lack of sensitivity to high cell density and the capacity to grow in low serum concentration, are dissociable from cellular tumorigeneicity. This conclusion is supported further by the demonstration that specific selection in vivo for tumorigenic cells from anchorage-dependent cells results in the isolation of anchorage-independent cells. Conversely, a single-step selection in vitro for anchorage-independent cells from nontumorigenic cells results in a simultaneous selection of highly tumorigenic subclones.


Deposition of histones onto replicating chromosomes. The mode of distribution of newly synthesized and pre-existing histones has been studied during the process of chromosome replication. Newly synthesized histone was labeled with [3H]lysine and newly synthesized DNA was density labeled with iododeoxyuridine. The histone was covalently linked to DNA, and radiolabeled histone was analyzed on CsCl density gradients. We have defined conditions that do not give rise to histone randomization during isolation, and also developed a method of defining the distribution of histones in chromatin on a density gradient in the unavoidable presence of nonhistone protein. Three possible modes of distribution of histone onto the replicating chromosome can be conceived; we describe experiments designed to distinguish unequivocally among these possibilities and conclude that histones are deposited randomly onto the chromosome.


Rous sarcoma virus activates embryonic globin genes in chicken fibroblasts. Complementary DNA (cDNA) specific for chick globin mRNA sequences fails to hybridize to total RNA extracted from chicken fibroblasts. After infection by Rous sarcoma virus, RNA complementary to globin cDNA is detectable in 100-500 copies per cell. Infection of fibroblasts with the transformation defective (td) deletion mutant of Rous sarcoma virus leads to normal virus production, but not to host cell transformation or accumulation of RNA sequences complementary to globin cDNA. Our evidence shows that the globin genes activated by Rous sarcoma virus are those specified by embryonic chick red cells; adult-specific globin sequences were not detected.


Cytochemical study of secretory process in transplantable insulinoma of syrian golden hamster. Electron microscopy, including phosphatase cytochemistry, indicates that the secretory granules of an insulinoma producing proinsulin and insulin are packaged by the endoplasmic reticulum (ER) and especially by a specialized region of ER which we call GERL because of the spatial relationship of this region to the Golgi apparatus and its apparent role in producing lysosomes. The granules are not derived from the Golgi apparatus. Preliminary evidence suggests this may be true also of pancreatic beta-cells.


Relationship in nucleic acid sequences between mouse mammary tumor virus variants. Primary cultures of mouse mammary carcinomas were used as a source of both radioactively labeled and unlabeled 60-70S RNA of mouse mammary tumor virus (MMTV) obtained from various mouse strains. Competition molecular hybridization experiments revealed that, within the limits of the assay, the RNAs of the MMTVs synthesized in culture by the tumors of the mouse strains RIII, GR, A, and C3H, are identical. A comparison of the genomes of the milk-transmitted MMTV(C3H) and the vertically transmitted MMTV(C3Hf) revealed that these two viruses are approximately 75% similar. No nucleic acid sequence homology was observed between MMTV(C3H) 60-70S RNA and the RNAs of murine leukemia virus, Mason-Pfizer virus, or the BrdUrd-induced type-B quinea pig virus.


Separation of sarcoma virus-specific and leukemia virus-specific genetic sequences of Moloney sarcoma virus. We have studied the nucleic acid sequences in nonproducer cells transformed by Moloney sarcoma virus or Abelson leukemia virus (two types of replication-defective, RNA-containing, viruses isolated by passage of Moloney leukemia virus in BALB/c mice). DNA probes from the Moloney leukemia in virus detect RNA in both Abelson virus-transformed nonproducer cells and Moloney sarcoma virus-transformed nonproducer cells. A sarcoma-specific cDNA, prepared from the Moloney sarcoma virus, has extensive homology to RNA found in heterologous nonproducer cells transformed by Moloney sarcoma virus, has little homology to RNA in cells producing Moloney leukemia virus, and no detectable homology to RNA in nonproducer cells transformed by the Abelson virus. By analogy to earlier data on avian and mammalian sarcoma viruses, these results suggest that the Moloney sarcoma virus arose by recombination between a portion of the Moloney leukemia virus genome and additional sarcoma-specific information, and indicate that the expression of this information in not essential for Abelson virus-mediated fibroblast transformation.


NMR in cancer: IX. The concept of cancer treatment by NMR: a preliminary report of high resolution NMR or phosphorus in normal and malignant tissues. A preliminary study of high resolution P31 nuclear resonance of normal and malignant tissue is reported. Utilization of nuclear resonance in the frequency frame contrasts with the pulsed magnetic resonance studies originally introduced for investigating cancer. Frequency dependent NMR was employed to study cancer with the primary objective of identifying resonant frequencies that could be used to indicate cancer. Such cancer specific frequencies would help circumvent current difficulties in the pulsed resonance diagnosis of cancer and might even have application in therapy.


Some aspects of ACTH action on cerebral metabolism in infant rats. In 150 newborn rats of either sex the influence of ACTH on cerebral phospholipid, phosphoprotein, and nucleic acids metabolism was investigated. The results show that ACTH induces a great stimulation of the cerebral phospholipid metabolism, especially in 14-and 21-day-old rats, a significant increase in the phosphoprotein metabolism at 7 and 14 days after birth, and a stimulation of the RNA synthesis in 7- and 14-day-old rats. The significance of these results is discussed.


Effects of 2200-m hypoxia on glucidic metabolism. Rats exposed for different intervals (1, 3 and 6 hours, the first day, and then 6 hours a day for 9 days) to a 2200-m hypoxia presented metabolic changes demonstrating an anaerobic glucose utilization. Thus, in accordance with other authors, at different depressurization values the liver homogenate had increased lactate and pyruvate concentrations, decreased NAD and NADH2 concentrations and a decreased ATP formation, demonstrating the inefficient glucide metabolization. These modifications were evident in the first hours of depressurization, but did not occur under acclimatization during the 9 days of exposure. The LDH concentration remained practically unchanged, which might be explained by its explained by its excessive presence in relation to the normal exercise solicitation. A urinary hypercompensatory alkalinization was also recorded.


Increases in paradoxical sleep as a result of amygdaloid stimulation. Holtzman rats were partially and selectively deprived of paradoxical sleep (PS) for a 16 1/2 hr period and allowed to rest for a 7 1/2 hr period each day. Test animals were given mild (60 Hz, 50 muA) unilateral amygdaloid stimulation for one min each day of a 5 day test period. There was a significant increase in PS during the rest  periods of test animals on stimulation days when compared to nonstimulated controls. The effect did not last beyond a 5 day period. Only subconvulsive behaviour was observed in two test animals. The similarity to the PS increases in this study to those seen in conventional learning and other stiuations are discussed.


Response of the adrenals to stress in animals from litters with different numbers of young. Undernutrition and overnutrition during the suckling period -- achieved by adjusting the number of infant rats per litter to 4, 8 and 14 -- resulted in a decrease in the plasma corticosterone level and in corticosterone production by the adrenals in vitro in overfed animals at the age of 30, 180 and 300--360 days. An enhanced response to ether anaesthesia or immobilization stress was found in overfed animals from the smallest litters. The results show that a change in postnatal nutrition and in social relationships in the litter influences maturation of the hypoathalamo-hypophysio-adrenal regulatory system; in adulthood this is manifested in changes in production of the adrenal hormones and in the reaction of the adrenals to stress.


Community mental health myths and the fate of former hospitalized patients. This paper addresses one of these important emerging problems--the fate of former hospitalized patients--and the policy issues that the problem raises. The discussion is derived from observations of the experience of the mental health system in the State of Hawaii, but the issues raised are of relevance to other states as well.


Factor analysis of objective and subjective characteristics of a night's sleep. Factor analysis was performed on objectively measured and subjectively reported characteristics of 46 nights' sleep in four healthy male subjects. The fragmentation of sleep by awakenings during the night, the delay before falling asleep, the total duration of sleep, and the amount of delta-wave sleep formed the bases of four separate factors.


Neuroendocrine effects of haloperidol therapy in chronic schizophrenia. The neuroendocrine effects of haloperidol therapy have been examined in 62 male chronic schizophrenic patients, aged 16-62 years. The duration of the disease varied between 2 and 29 years. The patients were divided into 48 hebephrenics with onset of the disease at puberty, or immediately after puberty, and 14 paranoids with onset of the disease in adulthood. They received 6 mg i.m.p.d. of haloperidol, for 30 days, up to a total dose of 180 mg. The following hormonal variables were examined before therapy and at 10-20 and 30 days of treatment: total urinary gonadotropins, serum FSH and LH, GH response to insulin stimulation, ACTH reserve (Metyrapone test), total urinary 17-ketosteroids and 17-hydroxycorticoids before and after an ACTH stimulation test, serum testosterone, insulin response to glucose load, plasma thyroxine before and after a TSH stimulation test. The basic hormonal values revealed decreased secretion of total gonadotropins, FSH, LH, ACTH and testosterone, and increased insulin secretion. The haloperidol therapy seemed to stimulate the secretion of FSH, LH, total gonadotropins, ACTH and testosterone, up to normal or low-normal levels. No modifications were observed in the other hormonal variables. The significance of these results is discussed.


Human sleep during chronic morphine intoxication. The sleep of 6 opiate addicts was studied for 11 nights during 3 phases of a chronic morphine cycle. The control phase consisted of 5 consecutive nights before morphine administration. The induction phase consisted of 1 night at 21-36 days after the onset of morphine administration, when the daily dose was 140-220 mg. The stable dose phase consisted of 5 consecutive nights after the subjects had received 240 mg of morphine daily for 8-19 weeks. No sleep could be studied during the withdrawal phase. Sleep was continuously monitored with EEG, EMG and EOG. Chronic morphine produces signs of a small but persistent sleep disturbance: delta sleep (early night) becomes less stable and shifts toward later in the night, waking state increases during the middle of the night, RME sleep (expecially its activated EEG without eye movements) decreases, the RMES cycle increases, and burst of delta activity (with mean duration of 5-6 sec) increase. Although this disturbance persists throughout the night, it is much less than that seen after single doses of morphine in a previous study. With chronic morphine, therefore, partial tolerance develops to the sleep disturbance produced by morphine. The small but persistent nocturnal arousal during chronic morphine contrasts with the sedation seen during chronic methadone. Both opioids produce an increase in delta bursts during chronic administration, which might be an EEG phenomenon specific to chronic opioid intake.


Studies of vitamin D deficiency in man.

Studies of vitamin D deficiency in man. Highly sensitive assays have been developed that enable 25-hydroxycholecalciferol (25-hydroxyvitamin D3) and 25-hydroxyergocalciferol (25-hydroxyvitamin D2) to be measured in the same serum sample. With these assays it has been shown that endogenously produced cholecalciferol (vitamin D3) is important in man; the findings further emphasize the role of vitamin D metabolites as hormones rather than vitamins in the traditional sense. Dietary sources of vitamin D appear to be inadequate and vitamin D deficiency has been shown to the cause of rickets and osteomalacia in Asian immigrants to Britain. This condition may be readily treated with small doses of vitamin D. In addition, sub-clinical deficiency was found in the Asian community. In the elderly, also, vitamin D deficiency was established as an important cause of osteomalacia and again evidence for the existence of a sub-clinical deficiency state was found. It is therefore suggested that the present prophylactic practices should be reviewed. Secondary hyperparathyroidism (reflected by elevated concentrations of circulating immunoassayable parathyroid hormone) was shown to be the rule rather than the exception in vitamin D deficiency. Some patients, however, had failed to respond to a hypocalcaemic stimulus. In others, there were high concentrations of parathyroid hormone despite normal serum calcium concentrations. Thus the relationship between parathyroid hormone and metabolites of vitamin D may not be mediated through changes in serum calcium alone, and it is postulated that metabolites of vitamin D may directly affect the secretion of parathyroid hormone.


Gastric ulcer and cancer.

Gastric ulcer and cancer. The incidence of gastric carcinoma was studied in a series of 210 patients presenting with apparently benign gastric ulcer. In eight cases (3.9 per cent), carcinoma was diagnosed within 18 months and was almost certainly present from the outset; in 2.4 per cent the diagnosis was delayed for over three months, and the five-year death-rate due to gastric carcinoma was 3.2 per cent. In a mean follow-up period of 5.7 years after the first diagnosis of an ulcer (8.2 years after first symptoms), three fresh cases of gastric carcinoma were found, and in two of these the cancer was at a different site from the ulcer. The five-year incidence of fresh gastric carcinoma was 0.6 per cent. Unsuspected superficial spreading carcinoma was detected by histology in 5.4 per cent of gastrectomy specimens. The significance of these findings is discussed in relation to the management of gastric ulcer and the early diagnosis of cancer.


Causes of death in fulminant hepatic failure and relationship to quantitative histological assessment of parenchymal damage. The clinical course and causes of death in 132 consecutive patients with fulminant hepatic failure and grade III or IV encephalopathy have been reviewed. 105 patients died and in 96 of these an autopsy examination was performed. In 36 patients there was cerebral oedema and the mean age of this group was significantly younger than the other fatal cases. In 28 patients death was attributed to major haemorrhage which originated in the gastrointestinal tract in 25. The prothrombin time ratio was not significantly greater in patients with major bleeding than in those without but they did have a significantly lower platelet count. Sepsis contributed to death in 12 patients. In 25 patients massive hepatic necrosis only was found at autopsy and death was considered to be due solely to hepatic failure. The degree of hepatocyte loss was assessed in 80 fatal cases by a histological morphometric technique on a needle specimen of liver taken immediately post-mortem. The proportion of the liver volume occupied by hepatocytes (hepatocyte volume fraction, HVF) was greatly reduced in all patients (normal 85+/-SD 5 percent) but the mean value was significantly higher in the patients dying with sepsis, cerebral oedema or haemorrhage than in the group in whom death was attributed solely to hepatic failure. There were ten patients in whom liver function was improving at the time of death which was due to cerebral (9) or haemorrhage (1). These observations suggest that many patients presently dying from fulminant hepatic failure may be expected to survive, once more effective therapy is available for the complications of the illness.


Heat-accelerated radioinactivation of attenuated poliovirus. Attenuated poliovirus is inactivated in a synergistic manner when exposed simultaneously to heat and ionizing radiation. The synergistic response is observed in both the thermally labile and stable forms of the virus. A three-term kinetic model may be used to describe the inactivation response of the virus in a thermal and/or ionizing radiation environment.


Triplet state population study in relation to the aggregation of proflavine in frozen solutions. The evolution of the proflavine triplet state population with the extent of aggregation is examines, by ESRmeasurements, as a function of organic solvents and mineral salts, well known for their ability to influence the aggregation process. The intensity of the triplet state population has also been investigated as a function of pH. The insertion of aromatic molecules like DNA's nucleotides between dye molecules is shown to increase the intensity of the triplet state. It is put forward that the observed triplet state is characteristic of the singly protonated proflavine which seems to be the active species in in vivo experiments.


Herpes genitalis in guinea-pigs. I. Kinetic study in infection with Herpesvirus hominis type 2. The kinetics of virus replication after vaginal infection of guinea-pigs with HVH 2/Angelotti were studied in relation to the appearance of local and general symptoms. Most virus was isolated from the genital tract 24-48 hours post infection. Virus was first isolated from the spinal cord 48-72 hours post infection. Penetration into the brain only occurred occasionally, and later. Under the experimental conditions employed, no virus was found in the blood, spleen, kidneys, adrenals or inguinal lymph nodes. The local symptoms (typical genital herpes) and the general symptoms (paralysis and death) started after maximum virus replication had been reached and seemed to be a consequence of neural, rather than of haematogenic or lymphogenic spread.


Herpes genitalis in guinea-pigs. II. Morphological studies in female guinea-pigs infected with Herpesvirus hominis type 2. Gross and microscopial morphological changes developing in female guinea-pigs after vaginal infection with HVH 2/Angelotti were studied. In the mucosa of the external genital tract there were inflammatory changes with formation of intra-epithelial vesicles, erosions and ulcerations. In the late stages of the infection signs of inflammatory dysplasia were also observed. The infection spread into the nervous system and produced characteristic inflammatory changes. The inflammation began as a bilateral posterior myelitis and ascended in the course of infection through the upper spinal-cord towards the brain-stem. The morphological changes were preceded by increased virus replication in the respective tissues and were correlated in time with clinical symptoms. The morphological changes seen at the site of inoculation in the external genital tract of the guinea-pig bore a certain resemblance to those seen in some cases of human infection with the same type of virus.


In vivo studies of parainfluenza I (6/94) virus: mononuclear cell interactions. Adsorption to and penetration of migratory mononuclear cells in vivo by Para-influenza I (6/94) virus were documented by electron microscopy during the acute phase of an experimental meningoencephalitis in ICR mice. Phagocytosis of virions was the apparent and predominant mode of viral entry into these cells. Penetration by fusion was also seen. Nucleocapsids were replicated by the mononuclear cells which became a potential reservoir for the further spread of virus.


Infection of guinea-pig embryo cells with varicella-zoster virus. Varicella-zoster (VZ) virus has been shown to replicate in guinea-pig embryo (GPE) cells. When cell-free virus inocula were used, no infectious virus was liberated from these cells, either by freezing-thawing or ultrasonic disruption. However, cell-free virus was separated from GPE cells inoculated with infected human embryonic lung (HEL) cells.


Experimental immunosuppression induced by herpes simplex virus. Cell-mediated immune response was studied in experimental herpetic infection in mice. Our data suggest that herpes simplex virus induced immunosuppression in mice. This depression was documented by the unresponsiveness of the lymphocytes to PHA stimulation in vitro and by the enhanced susceptibility of adult mice to Coxsackie B virus infection. Our data also suggest that levamisole, a potent immunostimulant drug, had no effect on the unresponsiveness of the lymphocytes to PHA stimulation.


Isolation of herpes simplex virus clones and drug resistant mutants in microcultures. Suitable dilutions of herpes simplex virus (HSV) preparations inoculated into microcultures of confluent monolayers of human foreskin or Vero cells, in individual wells of plastic "microplates", induced viral cytopathic effects that resulted from the infection of the cultures by single virus particles. The clonal nature of the viral progeny in isolated wells was supported by visual control over the development of viral foci and by statistical analysis. The method has the advantage of speed and economy, while it also yields a large primary clonal virus stock. HSV clones resistant to phosphonoacetic acid (PAA) and 5-iodo-2'-deoxyuridine (IUdR) could be readily isolated by the described technique.


[Neuroradiological study of traumatic cervical spine (author's transl)]. Only special neuroradiological procedures allow a clear demonstration or exclusion of a traumatic lesion involving or threatening the intraspinal structures. The myelographies are of particular interest. The functional sagittal pneumostratigraphy is the most informative technique, except for the visualization of an avulsion of the plexus. For better selecting the patients for a gas myelography, a scintigraphy of the spinal leptomeningeal bag is very helpful. But the neurological problem may be suspected on the standard X-ray picture. The lateral walls - articular apophyses and the arcus - are well defined. The vascular conditions of the nedk are best appreciated by an arch aortography. A selective study of the carotids and vertebral arteries in functional positions of the head and neck may then proceed.


[Early histological findings of carcinoma of the breast (author's transl)]. Histologically the early histological findings of the carcinoma of the breast enclose lobular and intraductal carcinoma in situ, and atypical epithelial proliferation. Without mastectomy a considerable percentage of these cases will develop invasive carcinoma. In order to detect these early forms of carcinoma mammography should be carried out. Following this procedure a more precisely located histological examination can be performed. The treatment consists of simple mastectomy.


Focal decreased skeletal uptake secondary to metastatic disease. Two cases are reported in which bone imaging with 99mTc-labeled phosphate compounds revealed localized areas of decreased uptake corresponding to osteolytic metastases on bone radiographs. These "cold" bone lesions, although infrequent, should be considered to avoid false negative interpretations.


Newer angiographic observations in cholangiocarcinoma. Three new angiographic features of cholangiocarcinoma are bile duct dilatation, venous obstruction and arterio-arterial collaterals. Differentiation of the lucencies of dilated bile ducts (cylindrical in shape) from those due to metastases (spherical) is discussed. Because of its high reliability and the difficulties of operative diagnosis, angiography can play an important diagnostic role in patients with suspected cholangiocarcinoma.


Inhibition of prostaglandin E2-induced cyclic amp accumulation in the rat anterior pituitary by 11-deoxyprostaglandin E analogs (9-ketoprostynoic acids). The effects of various 11-deoxyprostaglandin E analogs on the basal and prostaglandin E2 (PGE2)-induced cyclic AMP accumulation in the rat anterior pituitary were studied in vitro. 13-Hydroxy-9-oxoprost-14-ynoic acid at 5 X 10(-4)M but not 5 X 10(-5)M, decreased (45%) the induced accumulation and did not alter the basal accmulation; 15-hydroxy-9-oxoprost-13-ynoic acid at 5 X 10(-4)M caused less of a decrease (29%) in the induced and also did not alter the basal accumulation. (14Z)-13-Hydroxy-9-oxoprost-14-enoic acid at 5 X 10(-4)M did not alter the induced and caused a slight increase (5 fold) in the basal accumulation. 7-Oxa-13-prostynoic acid increased slightly the basal accumulation at 5 X 10(-5)M (2 fold) and 2.33 X 10(-4)M (6-fold) and did not antagonize the induced accumulation. Thus, the 9-ketoprostynoic acids are effective PGE2 antagonists in this system.


ACTH-induced prostaglandin biosynthesis from 3H-arachidonic acid by adrenocortical cells. Prostaglandins biosynthesized from 3H-arachidonic acid by trypsin-dispersed cat adrenocortical cells were isolated by silicic acid and thin layer chromatography. PGE, PGF, and a third component with mobility properties indistinguishable from either PGA or PGB were identified both in cortical cell homogenates and incubation medium. Concentrations of ACTH (125-250 muU) which stimulate steroidogenesis enhanced the conversion of labeled precursor to all three of these prostaglandins. These findings provide further evidence for the proposal that prostaglandins function as a critical link in ACTH-induced steroidogenesis.


Effects of indomethacin on intestinal secretion, prostaglandin E and cyclic AMP: evidence against a role for prostaglandins in cholera toxin-induced secretion. The effects of indomethacin on intestine mucosal cAMP, intestinal fluid secretion, and mucosal and fluid PGE were studied in rabbits in vivo following challenge with cholera toxin. Indomethacin had no effect on cholera toxin-induced fluid secretion or cAMP accumulation. Inhibition of PGE synthesis was achieved by the administration of two but not one injection of indomethacin. These studies provide evidence against a role for PGE in mediating cholera toxin-induced secretion and point out the need to measure prostaglandin levels when using prostaglandin synthetase inhibitors in vivo.


The relationship between prostaglandin release and lung c-AMP levels during anaphylaxis in the guinea-pig. A four-fold transient rise in c-AMP levels was seen when sensitized guinea-pig lungs were challenged with antigen in vitro. This rise in c-AMP also occurred in vivo and was shown to be due to release of Prostaglandin E2. This conclusion is supported by the finding that inhibitors of prostaglandin synthesis (Indomethacin and Poly phloretin phosphate) prevent the rise in c-AMP while neither ICI 74, 917, an inhibitor of histamine release, nor antihistamines had any effect on the c-AMP levels.


On the role of mitochondria in the hereditary cardiomyopathy of the Syrian hamster. Conventional polarographic techniques reveal no defect of oxidative phosphorylation in cardiomyopathic hamster heart mitochondria. However, in skeletal muscle of the same animals a defect is seen consistently with NAD+-linked substrates. This mitochondrial defect is caused by calcium accumulation and a consequent loss of NAD+ and magnesium. As shown by density gradient centrifugation, the defect resides in only a small fraction of all mitochondria. It is assumed that the same genetic defect in BIO 14.6 hamsters causes both skeletal muscle and heart lesions. Calcium levels in heart mitochondria are also significantly elevated in heart mitochondria. It is postulated that a calcium-associated defect similar to that in skeletal muscle mitochondria exists also in a very small fraction of the heart organelles and is responsible for the occurrence of focal necrosis.


Structural and functional characteristics of membrane fractions from cardiomyopathic and dystrophic muscle. Subcellular fractions from ventricular muscle of the cardiomyopathic Syrian hamster and breast muscle from the dystrophic chicken were prepared by zonal centrifugation in sucrose gradients. This tenchnique permitted the analysis of a wide spectrum of subcellular particles prior to sedimentation, thus avoiding loss of activity through particle clumping. Substantial differences in the distribution of beta-glucuronidase and p-nitrophenyl-phosphatase in zonal fractions from myopathic and control tissues provided evidence for contamination of fractions containing fragmented sarcoplasmic reticulum by lysosomes. This contamination may contribute signifantly to the altered lipid composition and Ca2+ transport function of the myopathic preparations.


Ultrastructural myocardial changes produced by viruses. Some of the ultrastructural pathological alterations in the myocardium produced by viral infections with members of the picornavirus group are described. These alterations varied from mild to severe, with necrosis. In animals infected with Coxsackie B1 virus and EMC virus, readily identifiable viral crystals were frequently found in association with the myocardial lesions by means of electron microscopy. The electron microscope made it possible to study vary early myocardial changes which were not detected with the light microscope. Although the ultrastructural myocardial changes in rheumatic fever have received little attention, the light and electron microscopic findings in the myocardium and valves of the viral-infected animals described here closely resemble those described in man in association with so-called rheumatic heart disease. The type of investigations presented forms a good model for the study of the natural history of ultrastructural pathology produced in all parts of the heart by viruses.


Virus-induced damage of the myocardial cell. A total of 330 male 3-week-old mice of the N.M.R.I./Han. strain were infected with the M-variant of the EMC (encephalomyocarditis) virus by subcutaneous inoculation of 0.25 ml of the virus suspension diluted 1:500. Animals were killed at 4- or 6-hour intervals, respectively, and groups of 2 to 8 hearts were taken for virological, morphological, and biochemical studies. Before the 8th hour after the inoculation, no virus and no pathological cell changes could be detected in the hearts; it is assumed, therefore, that there first takes place an extracardiac virus cultiplication. The virus content in the hearts increases from the 12th to the 30th ahour and subsequently maintains that level. Also from the 12th to the 30th hour, electron microscopy reveals increasing damage of the myocardial cells, the course of which is described. In most instances following virus-induced damage of the myocardial cells, the onset of alterations of the blood and lymph capillaries and damage to the interstitial cells were observed...


Spin labeled acetylcholine analogs: studies of cholinergic receptor. Some spin-labeled acetylcholine analogs, in which the number of methylene groups between the quaternary nitrogen and the ether oxygen ranged between 1-5, were synthesized to study drug interacitons with acetylcholine receptors. None of the compounds tested, with the exception of the one that contained 2 methylene groups (SL-2) had any cholinergic activity. SL-2 was not capable of producing any nicotinic cholinomimetic activity. On the other hand it proved to have a very weak nicotinic cholinolytic activity on the receptors of the frog satorius muscle. This compound exhibited strong antagonism against muscarinic receptors of the isolated frog heart. The muscarinic cholinolytic action of the spin-label ACh analog is discussed in terms of the molecular perturbation theory of drug action.


The growth characteristics of the metastatic Wistar/Furth Wilms' tumor model. Growth characteristics, metastatic spread, and survival times were evaluated in a transplantable Wistar/Furth rat Wilms' tumor model. Tumor suspensions of various tumor cell dosages were injected in an attempt to find the optimum level of tumor take. Lung metastases were frequent following tumor injection by all routes. However, tumor spread to the lungs was the least frequent following subcutaneous (SC) injection of the tumor. Survival time for the intramuscular (IM) group was statistically longer at all tumor dose levels. For the 1 X 10(5) tumor dosage, survival time ranged from a mean of 27 days for the intrarenal (IR) group to a mean 42 days for the IM group. For the 1 X 10(4) dosage, survival time ranged from a mean of 37 days for the intraperitoneal (IP) group to a mean of 51 days for the IM group. It is concluded that this animal tumor closely resembles the human Wilms' tumor, and that the point at which the transplanted tumor fails to be successfully transplanted is below the dosage level of 1 X 10(3) tumor cells.


Serum and host liver activities of glycosidases and sialyltransferases in animals bearing transplantable tumors. Rats bearing Reuber H-35 or Novikoff hepatomas and mice bearing L1210 or L5178Y murine leukemias exhibited elevated serum levels of fetuin : N-acetylneuraminic acid transferase (EC 2.4.99.1) activity. The serum transferase activity could be correlated with the growth rate of the tumor; in animals bearing the more rapidly growing Novikoff hepatoma, activity was higher than in animals bearing the Reuber H-35 hepatoma. Higher transferase levels were also found in L1210 leukemic mice than in mice with the slightly slower growing L5178Y leukemia. Serum from rats bearing Reuber H-35 hepatoma and mice bearing L1210 murine leukemia had elevated levels of alpha- and beta-glucosidase (EC 3.2.1.20 and EC 3.2.1.21), alpha- and beta-galactosidase (EC 3.2.1.22 and (3.2.1.23), beta mannosidase (EC 3.2.1.25), alpha- and beta-fucosidase (EC 3.2.1.- and EC 3.2.1.38), beta-N-acetylglucosaminidase (EC 3.2.1.30) and acid phosphatase (EC 3.1.3.2); alpha-mannosidase (EC 3.2.1.24), beta-N-acetylgalactosaminidase (EC 3.2.2.-) and beta-xylosidase (EC 3.2.1.37) were not elevated. In animals bearing Reuber H-35 hepatoma, host liver levels of glycosidases, beta-glucuronidase (EC 3.2.1.31) and acid phosphatase were elevated over both the control and the hepatoma values. The data are interpreted to mean that the tumors or various host tissues release large quantities of enzymes into the serum and that enzyme levels in host organs may also be affected by the tumor.


Effect of hydroxycobalamin on the inhibition of cytochrome c oxidase by cyanide. I - In intact mitochondria. The effects of hydroxycobalamin on inhibition of cytochrome c oxidase by cyanida in isolated intact mitochondria were studied. No effect of hydroxycobalamin (HCo) and cyanocobalamin (CNCo) was observed on normal mitochondria. When mitochondria inhibited by cyanida were treated with HCo the respiration with ADP, the respiration after ADP and respiratory coefficient (RC) were increased. No effect was observed with CNCo. Difference spectra of the effect of HCo on the inhibition of cytochrome c oxidase by cyanida and azide in isolated intact mitochondria were recorded using a split beam spectrophotometer. The results presented herein suggest that HCo reverses the cyanide inhibition because part of cytochrome a3 is free from cyanide as a consequence of dissociation of the complex, resulting in CNCo formation. If this reaction continues to occur together with a probable by pass between cytochrome a and oxygen throught the HCo (which was not transformed in CNCo), oxygen consumption and most of the respiratory chain will be maitened in the active state.


[Exocrine pancreatic function and calcium balance. Experimental studies of the effect of parathyroid hormone, vitamin D3, 25-hydroxycholecalciferol, dihydrotachysterin and thyrocalcitonin (author's transl)]. The effect of the acute elevation of extracellular calcium concentration, of vitamin D3, 25-hydroxycholecalciferol, dihydrotachysterin, parathyroid hormone and of thyrocalcitonin on the exocrine function of the perfused cat pancreas was studied. Elevation of the perfusate calcium concentration resulted in an initial increase in the enzyme and, occasionally, calcium concentration of the pancreatic juice, the flow rate and the secretion of magnesium and chloride were not very much altered. The application of vitamin D3 and 25-hydroxycholecalciferol elevated the stimulated (by acetylcholine or cholecystokinin-pancreocymin) protein and enzyme output (lipase, alpha-amylase). Parathyroid hormone and, especially, thyreocalcitonin inhibited the stimulated protein and enzyme secretion, volume and rate of the electrolyte secretion were not changed. In states of calcium excess the stimulated protein output was decreased by both vitamin D3 and parathyroid hormone.


Occupational type test for the etiological diagnosis of asthma due to toluene di-isocyanate. An occupational type exposure-test to toluene di-isocyanate (TDI) for the diagnosis of specific respiratory sensitization is proposed. The subjects were tested by exposure to a two-pot polyurethane varnish with and without TDI activator, using a paint-spraying system under standard and controlled conditions, so as not to exceed the threshold limit value of TDI. Forced expiratory volume per second and specific airway resistance (plethysmographic method) were measured before each exposure and during the following 24 h. 47 subjects with clinical symptoms of asthma were examined during their hospital care. For the purpose of this study they were divided into two groups: 42 workers exposed to TDI and 5 not exposed. The preliminary control test, using polyurethane varnish without TDI activator, was always negative and no significant changes in ventilatory parameters were recorded. The spray-painting test, with the same varnish and TDI activator, gave positive responses in 35 out of the 42 exposed workers and negative findings in the group not exposed. This method permits to formulate an accurate diagnosis of TDI respiratory sensitization in the majority of cases (83%). Immediate asthmatic reactions were frequently observed in the exposed subjects with high acetylcholine reactivity. Ten subjects showed a late bronchial reaction (3 h or more after TDI exposure).


Can VA/Q distributions in the lung be recovered from inert gas retention data? It has been suggested that the true distribution of ventilation-perfusion ratios can be determined from measurements made during the excretion of six different inert gases. This report shows that a number of considerably different distribution patterns will yield the same retention data, even in the absence of analytical error. Therefore, although analysis of inert gas retention data will allow one to arrive at a VA/Q distribution, it will not necessarily be the correct distribution.


[Heterochronia of nerve fibers in man. Diagnostic value: initial results]. A classical physiological observation not usually apparent with methods used for detecting nerve potential in man, heterochronia of the nerve fibres is often found in extraordinary circumstances with considerably higher than normal frequence : mainly in muscular disorders, in particular myotonic dystrophy ; more incidentally in disorders of muscle tone. The authors draw attention to certain sources of error which could lead to wrong diagnosis of heterochronia and emphasize that these are only the initial results of research which needs to be pursued further.


[Study of afferent conduction in the external saphenous nerve using the method of evoked cerebral potentials]. By the comparison of nerve action potentials and cerebral potentials evoked by the same electrical stimulus the propagation of the somaesthetic impulse to the cerebral cortex can be evaluated. This technique was applied to the fibres of the sural nerve in normal subjects and in patients presenting various lesions of the peripheral and central nervous system. Evoked cerebral potential afforded valuable data when it was not possible to record the nerve action potentials or when the lesion involved the central segment of the afferent pathway.


[Physiopathological and neuropathological study of a cerebral decortication syndrome]. Detailed observation of a case of laminar necrosis of almost the whole of the cerebral cortex has led to several deductions concerning the physiology of the cortex in man. It suggests that the cortex has a limited influence on sleep, interfering particularly in the electro-encephalographic translation of slow sleep and, curiously, in the organisation of sleep, paradoxical sleep is hardly affected at all. It confirms the importance of the human cortex in the acquisition of elementary skills and describes a new oculomotor pattern in a decorticate man.


Virologic studies in rheumatoid arthritis. Comparison of virus antibody titers in JRA and matched controls failed to implicate rubella, measles, parainfluenza type 1 or Epstein-Barr viruses. Neither RNA viruses or mycoplasma were detected in rheumatoid synovial cell cultures by 3H-uridine incorporation. Mycoplasma were not isolated from rheumatoid synovial membranes or cell cultures using the large specimen-broth culture procedure. The balance of available evidence, including our own, suggests that mycoplasma are not involved in RA. Viruses, however, are better candidates, in spite of similar generally negative studies to date, because methods for detecting known latent infection in animals and in vitro are still in their infancy. Those that are known have not yet been comprehensively applied to RA. Thus, supported by indirect evidence from animal models and human diseases like polyarteritis, the hypothesis that viruses are involved in the pathogenesis of RA remains attractive, although direct evidence is still lacking.


[Serological studies of the role of the respiratory syncytial virus in acute respiratory diseases in children]. The presence of the syncytial respiratory virus was determined by CF in 281 children admitted with acute respiratory diseases between 15 Sept. 1971 and 30 Dec. 1973, using the Long antigen prepared in the "St. Nicolau" Institute of Virology, Bucharest. In 38 children (13.5%) a serologic diagnosis of infection with the syncytial virus was established; in the other cases of respiratory infection of different etiology, antibodies to the syncytial virus were found in low but constant titers in both serum samples. The presence of these antibodies in a high proportion of the children points to the wide circulation of the syncytial virus in the infantile population, with all its clinico-epidemiologic implications.


Effects of chronic ethanol abuse on structure and enzyme activities of skeletal muscle in man. Biopsies from vastus lateralis muscle of male patients suffering from chronic ethanol abuse were studied with regard to histochemical reactions of ATPase and NADH-diaphorase; enzymatic activities of triosephosphate dehydrogenase (TPD), lactate dehydrogenase (LD), and cytochrome c oxidase (cytox); content of ATP, creatine phosphate, and glycogen; and volume fractions of fat, mitochondria, and fibrillar and extrafibrillar space. The results were compared with those from controls without known abuse of ethanol. The relative numbers of fibers were the same in two groups, but the size of the fast-twitch-glycolytic (white) fibers was diminished in the alcoholic group. The activities of TPD and LD were diminished in skeletal muscle of the alcoholics. This is most probably caused by the reduced amount of fast-twitch-glycolytic tissue, as there was a good correlation between this amount and the activity of the two enzymes. The activity of cytox was slightly lower in muscle of the alcoholics than in that of the controls. The volume fraction of mitochondria was lower in the alcoholic group than in the control group. Volume fractions of fat and fibrillar and extrafibrillar space were equal in the two groups. No significant differences were found in the amount of glycogen and ATP in the muscle of the two groups. However, the content of creatine phosphate is higher in the alcoholic group than in the control group.


Effect of treatment with a bile-sequestering agent (Secholex) on intestinal absorption, duodenal bile acids, and plasma lipids. Four female and five male patients (mean age 26 years) with hyperlipoproteinaemia type II A were treated with an anion exchange gel (Secholex) 9 g/day for 3 months and 15 g/day for 9 months. After these 12 months clofibrate 1.5 g/day was added to the therapy in 6 patients, whereas 2 patients continued with the resin alone for another 6 months, and one was withdrawn from the trial because of pregnancy. During the first year plasma cholesterol decreased averagely 18% from a mean pretreatment value of 461 mg/100 ml. Dosis of 9 g/day seemed to be as efficient as 15 g/day. When clofibrate was added, a further decrease of plasma cholesterol by 6% was observed, and the levels of triglycerides were reduced. Significantly increased concentrations of bile acids and a rise in the glycine/taurine ratio in duodenal aspirate were caused by the resin. On combined treatment the concentration of bile acids decreased to the pretreatment values, whereas the glycine/taurine ratio remained unchanged. During the trial slight transient changes in serum folic acid, fasting insulin, calcium, alkaline phosphatases, and vitamin B 12-absorption occurred. No changes in serum vitamin A, vitamin-K-dependent clotting factors, serum gastrin, gastric acid output, the absorption of glucose and iron, and faecal excretion of fat were observed. Serum insulin 30 and 60 minutes after an oral glucose loading decreased in the patients on combined treatment, whereas the insulin response remained normal in patients taking Secholex alone. Liver function tests and creatinine were unchanged during the trial. Apart from transient abdominal discomfort in two patients, no side-effects were discovered. The patients found the gel palatable.


Cell production and cell function in human cyclic neutropenia. In vitro studies have been done on haematopoietic cells from a patient with cyclic neutropenia characterized by severe depression of blood neutrophil levels every 21 days. Serial blood counts reveal periodic fluctuations in neutrophils, monocytes and reticulocytes. Agar culture of marrow cells shows normal concentration of colony forming cells. The percentage of colony forming cells in S phase is highly increased during profound neutropenia and normal during the recovery phase relating the granulocyte production to the peripheral neutrophil level. Studies of ingestion rate, bactericidal activity, lactate production and glucose oxidation during phagocytosis in isolated granulocytes show normal results. Also the ingestion rate in isolated monocytes is normal. Serial karyotype analyses of marrow cells during the neutrophil cycle display a normal pattern. Serum myeloperoxidase levels vary inversely with the peripheral neutrophil count indicating increased granulopoietic activity during profound neutropenia, which might be associated with non effective granulopoiesis during profound neutropenia, leading to a lack of granulocyte reserves in the marrow.


Stimulation of normal lymphocytes with autologous lymphoid cell lines: properties of derived killer cells. Lymphocytes from normal adults, with or without serological signs of previous Epstein-Barr virus (EBV) infection, could be stimulated to proliferate and produce killer cells by incubation with autologous EBV-genome-positive lymphoid cell lines (LCLs). The stimulated cells were most probably of T-cell origin, although at the peak of stimulation many of them lacked the sheep erythrocyte marker. Direct effector-target cell contact was necessary for lysis to occur. The cytotoxicity of autologously stimulated (AS) lymphocytes was not restricted to EBV-genome-positive LCLs, nor to cell lines of hematopoietic origin. It was equally broad if cells carrying complement receptor had been removed before stimulation. Fresh lymphocytes, blasts induced by phytohemagglutinin or concanavalin A, and Burkitt's lymphoma biopsy cells were resistant or considerably less sensitive. Mouse cells--even cell lines--were resistant. The sensitivity of target cells to lysis correlated positively with their capacity to block AS lymphocyte lysis of autologous LCLs in competition experiments. The cytotoxicity of AS lymphocytes was blocked by EBV-genome-positive and -negative cell lines, whereas the EBV-related cytotoxicity of T cells from acute cases of infectious mononucleosis was blocked by EBV-genome-positive LCL only.


Circadian periodicity a neurospora: alteration by inhibitors of cyclic AMP phosphodiesterase. Three methyl xanthine inhibitors of adenosine 3',5'-monophosphate phosphodiesterase-theophylline, aminophyline, and caffeine-lengthen the period of the circadian conidiation rhythm of Neurospora. The effect are seen in wild-type strains and in three mutant strains with genetically altered period lengths. These results suggest the possible involvement of adenosine 3', 5'-monophosphate in the control of circadian rhythmicity.


Immunosurveillance of naturally occurring feline leukemia. When compared to their housemates that subsequently developed leukemia, cats that remained healthy had five-to tenfold higher (geometric mean) humoral antibody titers to the feline oncornavirus-associated cell membrane antigen. This is compatible with the application of the immunosurveillance hypothesis to the natural development of leukemia in an outbred mammalian species.


Tumorigenicity of mouse-human diploid hybrids in nude mice. Somatic cell hybrids between normal mouse cells and simian virus 40 (SV40)-transformed human cells, which contained a diploid complement of mouse chromosomes and the human chromosome 7 carrying the genome of SV40, were tumorigenic in nude mice. One single copy of human chromosome 7 per hybrid cell appeared to be sufficient for the tumorigenicity of the hybrids.


Estrogen target sites in the brain of the chick embryo. Autoradiograms prepared from brains of chick embryos after injection of [3H]estradiol demonstrate the existence of target cells for estrogen in the medial preoptic and ventral hypothalamic regions as early as day 10 of incubation. Target cells also appear in telencephalic locations during later stages of embryonic development. These hormone-concentrating cells probably are the anatomical substrate for the formative action of sex steroids during embryonic life on certain brain functions such as control of sexual and aggressive behaviour and gonadotropin secretion.


Hepatocellular carcinoma: a clinical study. The clinical course of 40 patients with histologically proven hepatocellular carcinoma was reviewed. The majority had symptoms and signs suggesting abdominal malignancy; these included abdominal pain, weight loss, tenderness in the right upper quadrant, hepatomegaly, and fever. The most useful diagnostic tests were determination of serum alkaline phosphatase level, sodium sulfobromophthalein (Bromsulphalein) excretion, and liver scan. Prothrombin time and bilirubin levels were normal or only slightly elevated. Celiac angiography was helpful in determining the extent of the disease. Surgical exploration was done in 25% of the cases, but in only 5% was resection attempted. The grim prognosis is indicated by the fact that only 10% of patients were alive six months after admission to the hospital.


A study of serum lipoproteins and angiographic evidence of coronary artery disease. Extensive serum lipid analyses, with specific emphasis on the concentration of serum beta and pre-beta-lipoproteins, were made on 93 patients who had a clinical diagnosis of angina pectoris and were undergoing coronary angiographic studies. Those individuals with abnormal angiographic evidence of obstructive coronary artery disease, including a group with a prior myocardial infarction, have almost uniformly elevated serum beta-lipoprotein levels when related to an arbitrary division based on the mean values for a "normal" adult population at a similar age level. Compared to serum cholesterol and serum triglycerides, the serum lipoprotein concentrations, specifically beta-lipoproteins, appear to be a more useful parameter for predicting the existence of coronary artery disease. Further studies relating clinical risk factors to coronary artery disease might be better approached by using angiography of coronary vessels rather than myocardial infarction or sudden death from coronary artery disease as an endpoint of morbid data.


Localised myeloma with osteogenesis and Russell body formation. A case of an osteosclerotic myeloma of the mandible is described. Bone formation took the form of prominent sunray spiculation and radiologically mimicked an osteosarcoma. No other well-documented solitary lesion of this type could be found in the English literature. In addition, this tumour contained an abundance of intracytoplasmic Russell bodies and also produced a paraprotein. The paraprotein peak disappeared after resection.


Portal and pancreatic vein catheterization with radioimmunologic determination of insulin. Percutaneous transhepatic catheterization of the portal vein along with the catheterization of the aorta and the vena cava was performed in six patients, one of whom had hypoglycemic attacks due to Whipple's triad. Blood samples from different branches in the different vessel systems were withdrawn for radioimmunologic determination of insulin. By this method, preoperative localization of the insulin producing pancreatic islet cell tumor was su-cessfully performed. Multiple tumors as well as liver metastases were preoperatively excluded. Recatheterization and blood sampling for radioimmunologic determination of insulin was performed postoperatively to verify the radicality of the operation. Insulin concentrations from the hypoglycemic patients were compared with those of the patients without insulinoma and found to be significantly higher. The procedure is proposed as a useful method for localizing all types of gastrointestinal hormone producing primary or secondary tumors in which a method for hormone determination is available. The postoperative investigation is useful as a check of the operation performed and as a follow-up examination for the early diagnosis of recurrence.


Hepatocyte perfusion within a centrifuge. A centrifuge, as used in blood banks for washing erythrocytes, is shown to be a good perfusion chamber for hepatocyte suspensions being evaluated for potential extracorporeal liver support. The perfusate was cell-free oxygenated--pO2 300 millimeters of mercury--plasma, serum or tissue culture medium. A centrifuge that requires intermittent perfusion cannot deliver sufficient oxygen to maintain prolonged viability. A continuous flow unit can, at a flow of 800 milliliters per minute, support most functions of 100 grams of hepatocytes for six hours and some function for 12 hours. Metabolism of 2-14C tagged pyruvate to 14CO2 is the most useful sensitive measurement of liver function in this system. It is concluded that a centrifuge allowing a constant perfusion of oxygenated plasma into hepatocytes kept within the unit by centrifugal force is a suitable base for extracorporeal liver support.


Diagnosis of human fetal abnormalities by fetography. Being able to detect fetal abnormalities that may be associated with hydramnios would be extremely useful, especially when diabetes mellitus, Rh isoimmunization, and multiple pregnancy are ruled out. For this purpose the new technique of fetography, consisting of injecting a small amount of 2 radioque media (liposoluble and hydrosoluble), was used. Four out of 6 fetuses were correctly predicted to be abnormal. They were 1 case of esophageal atresia, 1 of suspicious chromosomal abnormality (after birth it was confirmed as having the Smith-Lemli-Opitz syndrome), and 2 of trisomy 18. It is felt that this simple technique should be used as an aid to the obstetrician faced with the problem of determining the basis of unexplained hydramnios.


Electron microscopic observation of inclusion bodies in plasma cells of multiple myeloma and Waldenström's macroglobulinemia. Intracellular inclusion bodies in the plasma cells were sought by electron microscopy in 32 cases of multiple myeloma and 3 of Waldenström's macroglobulinemia. In some cases, round shaped intranuclear inclusions and intranuclear fibrillar bundles were observed. In other cases, Russell bodies and crystalline structures were found in the cisternae of rough surfaced endoplasmic reticulum (rER), and dense bodies, myelin-like structures, fibrillar formations, polysome lamellae complexes, crystalline structures, virus-like particles and phagocytosis were observed in the cytoplasm of plasma cells. The detailed ultrastructure of these inclusions was described, and their functional significance, origin and appearance rate were discussed. Finally, the presence of true viral particles in the plasma cells was ruled out.


(3H)-estradiol binding by chick liver nuclear extracts: mechanism of increase in binding following estradiol injection. Specific high affinity binding of [3H]-estradiol by 0.5 M KCl extracts of chick liver nuclei is substantially increased by estradiol injection of the immature chick. The effect is observed shortly after estradiol injection, while the estradiol-induced production of serum phosphoproteins (vitellogenic response) is not detectable until about 24 hr. Cycloheximide given 90 min before estradiol inhibits the increase in nuclear binding for 12-15 hr. At 24-48 hr the levels of nuclear binding are similar to those in the estradiol-treated animals not given cycloheximide, but serum phosphoprotein levels are depressed by about 80% at 48 hr. By 75 hr however the serum of the cycloheximide-treated estrogenized chicks contains about twice as much phosphoprotein as does serum of chicks given estradiol alone. It is suggested that the inhibition of protein synthesis for 12-15 hr delays the vitellogenic response until sufficient levels of nuclear [3H]-estradiol binding protein can be synthesized. A correlation between the levels of nuclear [3H]-estradiol binding at 24 hr and phosphoprotein at 48 hr is shown in a dose-response experiment. In vitro, nafoxidine-HCl (Upjohn 11,100 A) inhibits binding of [3H]-estradiol by the chick liver nuclear extracts. In vivo, a single injection of nafoxidine with estradiol inhibits phosphoprotein production. Injection of nafoxidine alone results in a small but significant increase in [3H]-estradiol binding by nuclear extracts, but it is not estrogenic. A possible interpretation is that nafoxidine transfers low levels of a putative cytosol receptor to the nucleus, but is unable to induce the amplification mechanism required to give the levels of nuclear estradiol-binding protein needed for the vitellogenic response.


Estrogen receptor in rat liver and its dependence on prolactin. Estrogen receptor is shown to be present in the livers of adult rats. The receptor binds estradiol-17beta with a Kd of 1 x 10(-10) M and sediments at 8 S in sucrose gradients. Other estrogens and anti-estrogens compete for estradiol binding, while nonestrogenic steroids do not. Receptor levels fall dramatically after hypophysectomy, but can be partially restored within 18 hours by a single injection of prolactin. It is known that prolactin critically regulates the level of its own receptor in the liver, and we now suggest that it also exerts a primary control over the availability of liver estrogen receptor.


Metabolism of dehydroepiandrosterone by hormone dependent and hormone independent human breast carcinoma. The metabolism of 7alpha-3H-dehydroepiandrosterone was studied in six human breast carcinomas in vitro. All mammary tumors transformed DHA to testosterone, dihydrotestosterone, 5alpha-androstane-3alpha, 17beta-diol and 5alpha-androstane-3,17-dione. Of the tumors investigated, three estrogen receptor-negative tumors converted DHA to estradiol and only one estrogen receptor-positive tumor produced estradiol from DHA. Observations on the relationship of androgen metabolism and hormone dependency are discussed.


[Experimental and clinical evaluation of stomatological cleaning agents with special reference to the Eladent prosthesis cleaner]. The experimental and clinical testing of certain denture cleaners, viz., Eladent, Kukident and Steradent, yielded identical good results. As far as possible, Eladent should be dissolved in warm water (35-40 degrees C). In this case, the tablets will dissolve within 15-20 minutes. For immediate cleaning, the tablets may be dissolved within 1 or 2 minutes by raising the water temperature at 50 degrees C. No striking immediate effect can be expected from this cleaner in case of stained dentures which have been worn for many years. The cleaner improves little by little the hygienic condition of the denture. It is recommended to apply the cleaning tablets immediately after insertion of the prosthesis at regular intervals. In this way, soft deposits and plaques will no longer adhere and cause the formation of hard deposits.


Physiological properties of vertebrate nerve cells in tissue culture. Vertebrate neurons in tissue culture are providing us with a new model system for studying the complex events which occur during neuronal differentiation, synaptogenesis, and neural network formation. It is already apparent that dissociated embryo neurons are capable of differentiating both morphologically and physiologically along predetermined lines in the absence of external influences. These neurons can form new connections with one another but retain some specificity in their selections. Both simple and complex neural networks can be seen. At the present time, the development of the invitro model system is just being explored. The potential value of a system of this kind at a variety of investigative levels should be appreciated. Questions of a fundamental nature in neurobiology, such as how synapses form, what rules govern such interaction, how cells recognize one another, and the nature of the basic two-, three-, or four-cell circuits that comprise the more complex neurons tissue can be approached with this system. Studies of the neurons and synapses themselves can lead to a more basic understanding of vertebrate nervous system functioning. The development of certain pathophysiological processes and the effects of neuroactive drugs on vertebrate neurons may be studied at the cellular level. Finally, the basic mechanism of some genetic abnormalities which produce abnormal nervous structure and function may be more easily determined in a simplified in vitro model than in the intact central nervous system. The value of any model is not inherent in the elegance of the model itseld, but only in its ability to suggest answers to fundamental questions about the system being modeled. Many fundamental questions about brain mechanisms in mental retardation remain unanswered. Perhaps some day the model of nerve cells in tissue culture will bring us closer to the answers to these questions.


Studies on the molecular defect in galactosemia. The galactose metabolic pathway and some of the consequences of deficient galactokinase or gal-1-P uridyltransferase activity have been discussed. The existence of CRM in transferase deficiency galactosemia is presented as evidence that this disease is the result of a structural gene mutation. The finding of both quantitative and qualitative variation in transferase CRM among different galactosemic patients argues that genetic heterogeneity exists within this group. Data supporting a Ping-Pong mechanism of action for human transferase reaction is proposed.