[Experimental and clinical study test of capreomycin]. Data on the experimental and clinical study of capreomycin in the treatment of tuberculosis are presented. It was shown that capreomycin had low activity with respect to the sensitive strain of Mycobacterium tuberculosis H37 Rv in vitro and the respective infection caused by it in mice. The activity of capreomycin in vitro with respect to streptomycin resistant strains was the same as that with respect to the sensitive strains, while in vivo it increased 3 times. Capreomycin showed a tendency to decreasing its activity with respect to strains highely resistant to canamycin only in vitro. The effect of capreomycin on tuberculosis infection caused by strains resistant to different concentrations of canamycin was the same as that on tuberculosis infection caused by sensitive strains. Cross resistance between florimycin (viomycin) and capreomycin was shown. Clinical trails of capreomycin revealed its moderate therapeutic efficiency, relatively low toxicity and an allergenizing effect on the host. Transient ventibulopatia without pronounced signs of ototoxic action was observed. The nephrotoxic effect was moderate and transient. It was observed predominantly at the peak of the allergic reactions to the antitubercle drugs. The data obtained during the study allow recommendation of capreomycin use in clinics as reserve drug when the causative agent is not resistant to florimycin. The drug should be used under regular control of the blood picture, electrolyte metabolism, state of the kidneys, auditory and vestibular apparatus.


A colour reaction for the differentiation of ascomycetous and hemibasidiomycetous yeasts. Seventy yeast strains, representative of twenty-six ascogenous genera, four saprobic hemibasidiomycetous genera and thirteen genera of the Cryptococcales were tested for their reaction with the stabilized aromatic diazonium compound, Diazonium Blue B salt. An aqueous, buffered solution of this compound gave a characteristic red colouration with the colonies of the hemibasidiomycetous species and those Cryptococcales characterized by the hemibasidiomycetous cell-wall type. The characteristic colour reaction was not observed with colonies of either the ascomycetous yeasts or those Cryptococcales characterized by the ascomycetous cell-wall type. The possibly taxonomic use of the colour reaction with Diazonium Blue B salt as an affinitive characteristic is discussed.


Staining of minerals and solubility of iron in tissues. Iron deposits in ethionine induced aortic siderosis of rats, in splenic deposits in sickle cell anemia and siderocalcific vessels in cerebral arteriosclerosis were completely removed by exposure to 20 percent hydrochloric acid for 30 min. This contrasted with idiopathic hemochromatosis and idiopathic pulmonary hemosiderosis in which the iron containing organs had to be exposed to 40 percent hydrochloric acid for two hours. The more soluble iron appeared colorless in unstained tissues, purplish blue with hematoxylin and eosin, turquoise blue with Perls' stain, violet blue with gallocyanin and dark-drown with sodium rhodizonate. The less soluble iron was golden yellow in unstained tissues. It appeared golden yellow with hematoxylin and eosin and sodium rhodizonate, but it stained greenish blue with Perls' method and dark brown with gallocyanin. Lead and copper were capable of deposition in some tissues in vitro in the presence of iron and/or calcium but not when these minerals were removed. This phenomenon may be of importance in certain pathological conditions, e.g. hemochromatosis, where on preexisting tissue-iron-complexes there is a secondary deposition of copper.


[Additive activity of IgE and IgGa anaphylactic antibodies in rats (author's transl)]. Anaphylactic antibodies of two different classes are produced in rats: IgE and IgGa. When injected into the skin of normal animals, IgE remains at the site of injection for two weeks or more, while IgGa diffuses out within hours. In order to determine whether the anaphylactic reaction produced by one class of antibody is modified by the simultaneous presence of the other class of antibody, sites were prepared in normal rats with IgE antibody and 24 hours later IgGa antibody was injected into the same sites. The passive cutaneous anaphylactic reactions was elicited 2 hours later by intravenous challenge with an excess of antigen. The results indicated that both antibodies acted in an additive way. No inhibition or potentiation was observed.


Cross-sensitivity of common aminoglycoside antibiotics. Guinea pigs were sensitized to neomycin (A, B, or C), paromomycin, gentamicin, kanamycin, streptomycin, and dihydrostreptomycin via intradermal or foot-pad injection with an adjuvant containing killed Mycobacterium butyricum or M tuberculosis H37Ra (Ra). These antibiotics produced greater cross-sensitization with an increase in the number of immunizations and chemical structural similarities. After repeated intradermal injections (adjuvant Ra) of neomycin, guinea pigs showed cross-sensitization to paromomycin, kanamycin, and streptomycin. A single intradermal injection of one of these antibiotics produced stronger reactions to the most closely related antibiotics, with no meaningful sensitization to the least-related allergens. Streptomycin-sensitized guinea pigs seldom showed a meaningful cross-sensitization to dihydrostreptomycin or the other antibiotics (except neomycin C); however, guinea pigs sensitized to dihydrostreptomycin or the other antibiotics often showed strong cross-sensitization to streptomycin.


Predictive value of early developmental examination. A developmental examination was carried out on 168 children at intervals over a 3-year period. The results suggest that developmental screening of this type is effective in picking out the "poor performers" who might well benefit from preschool education and extra health visitor supervision. On the other hand, the test given to children under the age of one year failed to pick out 2 children who were found to be moderately handicapped at the time of the 3-year examination. The study under-lined some of the problems of universal screening which require further research, and the need to assess the results of developmental screening clinic.


Chlorpromazine-induced histamine release from guinea-pig skin in vitro--a photosensitive reaction. Chlorpromazine releases histamine from chopped guinea-pig skin maintained in vitro at 37 degrees C. This effect was markedly potentiated by radiant energy. This potentiation increased with increasing exposure to radiant energy. The possible clinical applications of these findings are discussed.


Attempts to identify viruses in rheumatoid synovial cells. Synovial fibroblast cell strains derived from the synovial membranes of 7 patients with rheumatoid arthritis were examined for the presence of viruses, in particular leucoviruses. Seven similar synovial strains derived from patients with other arthritic conditions were used as a control group. Evidence of the presence of a virus or a viral genome was looked for by several methods of induction followed by 3H-uridine labelling of the cultures. In addition, the culture supernatant, after induction and after the synovial strains had been co-cultivated with a variety of cell lines from several species, was assayed for the presence of viral RNA-dependent DNA polymerase activity. The DNA-polymerase activity of the synovial cells themselves was also determined. No evidence was found by any of these techniques to indicate the presence of virus or viral information within the synovial fibroblasts.


The pharmacology of butorphanol, a 3,14-dihydroxymorphinan narcotic antagonist analgesic. Butorphanol, a new, totally synthetic morphinan, which is chemically related to naloxone, has been demonstrated to have both analgesic and narcotic antagonist properties. In rodent antiwrithing analgesic tests, butorphanol was 4 to 30 times more potent than morphine and dl-pentazocine, respectively. As an antagonist, butorphanol was about equivalent to nalorphine and 30 times more potent than dl-pentazocine. On the basis of the naloxone-induced mouse jumping test and the lack of substitution in withdrawn morphine-dependent mice, it is estimated that the potential for physical dependence of butorphanol will be less than that of dl-pentazocine but greater than that of nalorphine and dl-cyclazocine. Animal data also show that agonistic actions of butorphanol, such as respiratory depression and miosis, reach ceiling effects which are lower than those seen with morphine with an increase in dosage. Thus, butorphanol differed from morphine which exhibited agonist effects in a dose-related manner. Butorphanol showed weak to moderate central depressant properties at doses which were considerably higher (greater than 100 X) than those producing analgesia. Minimal cardiovascular and respiratory effects were seen with butorphanol in conscious dogs.


Utilization and degradation of lindane by soil microorganisms. Of 147 microorganisms isolated from a loamy sand, 71 showed good growth with lindane (gamma-1,2,3,4,5,6-hexachlorocyclohexane) and produced chloride in an aqueous medium. Thirteen soil microorganisms were selected to study the utilization of lindane. Lindane was metabolized by the microbes to gamma-2,3,4,5,6-pentachloro-1-cyclohexene (gamma-PCCH), alpha-3,4,5,6-tetrachloro-1-cyclohexene (alpha-TCCH), beta3,4,5,6-tetrachloro-1-cyclohexene (beta-TCCH), gamma-3,4,5,6-tetrachloro-1-cyclohexene (gamma-TCCH), and pentachlorobenzene (PCB). Cells of Pseudomonas sp. No. 62 grown on lindane simultaneously adapted to gamma-PCCH, alpha-TCCH, beta-TCCH, gamma-TCCH, PCB, 1,2,3,4,-tetrachlorobenzene (1,2,3,4-TCB) and 1,2,4,5-tetrachlorobenzene (1,2,4,5-TCB). The bacteria degraded each of these chemicals at least partially as indicated by an increased rate of oxygen consumption.


Evaluation of propranolol in opiate dependence. Among the increasing number of recent reports of the use of beta-adrenergic blocking drugs for a variety of psychiatric disorders is a series concluding that propranolol hydrochloride is efficacious in the treatment of opiate dependence. These reports were based on clinical observations of outpatient addicts in an uncontrolled situation. In order to validate these findings, we carried out a series of controlled, double-blind studies with hospitalized volunteers. The results of our studies do not confirm the previous reports. Propranolol neither relieved nor precipitated opiate withdrawal in subjects dependent either on dihydromorphinone hydrochloride or on heroin. It did not effectively block heroin-induced euphoria in dependent or nondependent subjects. In addition, there was no effect on the pattern of self-administration in opiate-dependent subjects, whether they were maintained on propranolol or placebo.


Histologic fixatives suitable for diagnostic light and electron microscopy. The merits of formaldehyde, formaldehyde-glutaraldehyde combinations, and glutaraldehyde in phosphate buffers have been compared as fixatives that will give easy and satisfactory preservation of tissues for routine automated histologic processing and yet keep them suitable for electron microscopical studies after prolonged storage at room temperature. We recommend a combination of 4% commercial formaldehyde and 1% glutaraldehyde in a buffer of 176 mOsm/liter. Tissue sections should not exceed 3 mm in width, and tissues to be examined by electron microscopy should be taken from the outside of the tissue sections. All special stains performed at light microscopical level gave satisfactory results. The fixative should be stored at 4 C and is stable for at least three months. The superior cross-linking features of glutaraldehyde are retained, while the concentration of glutaraldehyde is low enough not to substantially obscure the PAS reaction.


A histological stain for detecting Friend leukemic cells in fixed hemopoietic tissue. A histological stain described by Menzies in 1963 has been useful in detecting early evidence of erythroleukemia (Friend disease) in mouse hemopoietic tissues fixed in formaldehyde solution. With the use of this stain, one can distinguish erythroblastic leukemia cells from lymphoblastic leukemia cells.


Therapeutic oophorectomy in disseminated carcinoma of the breast. Of 145 women with metastatic breast carcinoma who survived palliative surgical castration, 40.7% had objective remission and another 11.0% had subjective improvement. Favorable response was more frequent in ovulating patients who had small masses of recurrent tumor in fewer locations, with skeletal spread especially well tolerated. Palliation appeared to be related to the site of metastatic disease and numbers (mass) of metastatic sites, but not to the age of the patient, tumor histology, or the disease-free interval. Survival was better for the patients who showed improvement after castration. Such improvement may predict continued relief from the use of other modalities of systemic treatment.


Chalcosis in the human eye. A clinicopathologic study. A clinicopathologic study of eight human eyes containing intraocular copper foreign bodies was undertaken. These foreign bodies, which had been retained in the eyes from three weeks to 22 years, were analyzed by atomic-absorption spectrophotometry and x-ray diffraction to determine their chemical composition. Copper deposits in Descemet membrane, vitreous, internal limiting membrane of the retina, and fibrous capsule around the foreign bodies could be demonstrated by rhodanine, rubeanic acid, and alizarin blue stains and were correlated with clinical findings and pathologic changes. Eyes with foreign bodies containing more than 85% copper tended to show more disseminated copper deposits. Eyes with alloy containing less than 85% copper showed more localized deposits of copper. Retinal structures were well preserved, even in an eye with an intraocular copper foreign body that had been retained for 22 years.


[Reactions and damage following external influence of drugs and occupational noxae on the skin]. Changes of the skin by the external influence of drugs and occupational substances are to be divided up into direct toxical influences, deposits, pharmacodynamical effects and immunologically directed reactions. With easily penetrating substances serious general effects like the formation of methaemoglobin and the intoxication with mercury have been observed. Development, diagnostic, prophylaxis and therapy of the cellular immunological reaction are thoroughly set forth.


Influence of the adrenal glucocorticoids on the stimulation of synthesis of hepatic ribonucleic acid and plasma acute-phase globulins by leucocytic endogenous mediator. An injection of unpurified leucocytice endogenous mediator into rats results in an increased incorporation of [6(-14)C]orotate into hepatic RNA, an increase in the concentration of RNA associated with the bound ribosomal fraction of liver, and increases in the concentrations in serum of acute-phase proteins such as  alpha2-macrofoetoprotein and haptoglobin. If given 3 days after adrenalectomy or 7 days after hypophysectomy,, leucocyte factor did not induce the increase in RNA synthesis or alpha2-macrofoetoprotein concentrations but did stimulate an increase in serum haptoglobin. When hypophysectomized or adrenalectomized rats received daily subcutaneous injections of 0.5mg of cortisol, leucocyte factor again induced a significant increase in the synthesis of hepatic RNA and an increase in the concentration of serum alpha2-macrofoetoprotein. These observations suggest that leucocyte factor can regulate acute-phase-protein synthesis at several different sites, one or more of which requires permissive action of the glucocorticoid hormones. Futher, leucocyte factor will stimulate an increase rate of incorporation of orotate into hepatic ribosomes when added in vitro in the presence of cortisol to a liver-perfusion system. Thus the stimulatory effect of leucocyte factor may be directy on liver but may require the presence of other hormones to stimulate the incorporation of orotate into RNA.


Mononuclear cells in human synovial fluid. Identification of lymphoblasts in rheumatoid arthritis. A simple method was developed to identify large mononuclear (LMN) cells in human synovial fluid based on morphology and staining with Sudan black B. All cells were classified as monocyte-derived macrophages (MDM), lymphoblasts (LB), or synovial lining cells (SLC). Lymphoblasts were seen in 58 of 60 rheumatoid fluids (mean: 69 +/- 18% LB per LMN cells). However lymphoblasts were rarely seen in synovial fluids from patients with crystal-induced synovitis or bacterial infections.


P-component of amyloid. Isolation from human serum by affinity chromatography. Human amyloid P-component was isolated from plasma by affinity chromatography utilizing antiserum to tissue P-component coupled to Sepharose 4B. Characterization and comparison of the isolated P-component proteins from tissue and plasma demonstrated immunologic identity and identical RF values of polyacrylamide disk gel electrophoresis. Amino acid analyses of P-components from two individual plasmas were comparable, but some variations from tissue P-component were noted. Attempts to perform amino acid sequencing on the plasma protein were unsuccessful. Electron microscopic studies revealed that the plasma P-component had a pentagonal ultrastructure identical to the tissue P-component. These studies compared similarities of tissue and plasma P-components and revealed the unique pentagonal ultrastructure of the plasma protein.


Clinical assessment of the MOD-MEM cancer test in controls with non-malignant diseases. A control series of 105 patients in hospital with non-malignant diseases was used in a limited clinical assessment of the MOD-MEM test. Twenty-seven positive results could be explained on the basis of destruction of nervous parenchyma, tissue necrosis, tuberculosis, malignant disease, etc. The remaining 13 unexplained positives showed a sex and age distribution in agreement with that predicted from cancer registration statistics if the MOD-MEM test detects cancer about 16 years before the clinical appearance of the disease.


Responses of cancer patients in the MEM test: not just a function of charge on basic proteins. It has been reported that lymphocytes from cancer patients give positive responses to PPD, myelin basic protein, tumour basic protein, and certain histone fractions in the MEM test. The underlying mechanisms of the MEM test are poorly understood, but it is widely assumed that it detects immunological sensitization to specific antigenic determinants. The cross-reactivity experienced is interpreted as indicating shared antigenicity. Since all the stimulatory proteins are strongly basic we investigated an alternative explanation that responsiveness is a function of electrical charge by comparing the known stimulatory proteins in the MEM test with two others of similar basicity: lysozyme and cytochrome-C. We obtained highly significant stimulation with PPD, tryptophane peptide of myelin, and tumour basic protein using Mantoux + cancer patients, but found no response to other basic proteins. We failed to confirm the reported activity of histone F2a. Our results indicate that basicity alone is insufficient to elicit response, and strengthens the concept that the MEM test is measuring sensitization to the determinants shared by myelin and tumour basic protein.


Anti-streptococcal antibodies reacting with brain tissue. II. Utrastructural studies. The reaction of rabbit anti-Streptococcus pyogenes type 24 with mouse brain has been studied at the ultrastructural level using horseradish peroxidase labelled antiglobulin. It is shown that these antibodies, and by inference those found in Sydenham's chorea, react with the fibrous astrocyte. The cross-reacting antigen is the glial filament or a closely associated cytoplasmic constituent.


Kinetics of in vitro bovine keratohyalin synthesis. A system is described for the study of the in vitro synthesis of bovine keratohyalin for periods of up to 24 h. Keratohyalin granules appeared morphologically unaltered during culture, although histochemical stains for RNA indicate a markedly diminished RNA content by 6 h. Incorporation of tritiated histidine began slowly, then became linear between 4 and 24 h in serum-containing media. However, following pre-incubation in serum-free media, increased incorporation occurred from time zero. The internal variation of the system using a standard 6 h pulse was +/- 10, +/- 24, +/- 22, +/- 18% in four separate experiments. Measurements of modulations in bovine keratolhyalin synthesis in vitro may prove useful in the development of drugs of therapeutic potential in dermatology.


Different antiplatelet antibody specificities in immune thrombocytopenic purpura. It is generally accepted that patients with immune thrombocytopenic purpura (ITP) produce antibody against platelet-associated antigens; however, it is not known if these antiplatelet antibodies are directed towards the same or different antigenic sites. In the present studies, quantities of antiplatelet antibody from different ITP patients, sufficient to saturate platelet antigenic sites, were simultaneously incubated with normal platelets and the quantity of platelet-binding IgG (PBIgG) was determined. In each of the five comparisons made, the amount of PBIgG bound after incubation of normal platelets with saturating quantities of two ITP antibodies approximated to the sum of the PBIgG bound after incubation with the antibodies separately. These data suggest that the antiplatelet antibody from these ITP patients differed in antigenic specificity.


Defective neutrophil function in chronic granulocytic leukaemia. The ability of neutrophils to phagocytose, kill and digest Candida guilliermondii was investigated in twelve patients with chronic granulocytic leukaemia (CGL). Following ingestion of organisms there was considerable reduction in the ability of CGL cells to kill Candida and this was not explained by a mild impairment of phagocytosis. Histochemical staining showed that granules containing lysosomal enzymes disappear form the cytoplasm of normal neutrophils during killing and digestion of the fungus, while in CGL cells the granules remain. Quantitative measurement confirmed loss of peroxidase from normal neutrophils during Candida killing but no loss from CGL neutrophils. The primary granules of CGL neutrophils do not fuse with and discharge their contents into the phagocytic vacuole and this explains their impaired ability to kill and digest Candida.


Maternal serum alpha-fetoprotein in normal pregnancy. Serial measurements of alpha-fetoprotein (AFP) were made in 63 normal pregnancies throughout gestation using a radioimmunoassay. Significant levels of AFP, which were detected as early as at 10 weeks gestation, showed a gradual rise until 32 weeks, after which the levels declined until term. The limits of the normal gestational pattern have been defined and the intrapatient variability of the assay was examined. A comparison of the clinical characteristics of the pregnancies with levels of AFP was made. Higher birth weights were correlated with later attainment of peak levels of AFP and peak levels were achieved earlier in gestation in female fetuses. Higher levels of AFP and lower oestriol/AFP ratios were correlated with an earlier onset of labour.


HCG, HPL, oestradiol, progesterone and AFP in serum in patients with threatened abortion. The predictive value of various biochemical methods for monitoring early risk pregnancies has been compared in 65 cases of threatened abortion. Estimation of human chorionic gonadotropin (HCG), human placental lactogen (HPL), progesterone, oestradiol and alpha-fetoprotein (AFP) in serum were made by radioimmunoassays. Values below the normal range predicted abortion in 79, 81, 89, 92 and 38 per cent of patients, while normal values confirmed continuation of pregnancy with an accuracy of 71, 61, 60, 68 and 30 per cent respectively. Thus predictions from oestradiol and progesterone were at least as reliable as those from the protein hormones, while AFP proved to be unsuitable for this purpose. Combination of two variables gave even more reliable results. Due to individual and diurnal variation, however, abortion in the third and fourth month could not be definitely assumed at values above 5 IU HCG/ml, 5 ng progesterone/ml or 200 pg oestradiol/ml.


The relationship between HLA antibodies and the causal pregnancy in choriocarcinoma. By studying the HLA system, evidence of immunization by paternal antigens inherited by the antecedent child has been obtained in primiparous patients who subsequently developed choriocarcinoma. The same approach has been used to show which pregnancy was likely to be casual in two patients with complicated obstetric histories.


Preparation and properties of O-dansyltyrosine gramicidin C. Gramicidins A, B, and C are a family of poly-peptide antibiotics which facilitate the passive diffusion of alkali cations and protons through lipid bilayer membranes. It is clear that gramicidin forms a multimeric transmembrane channel and it has been suggested that the channel is an io-conducting dimer in equilibrium on the membrane with non-conducting monomer. We describe the preparation and purification of a derivative of gramicidin C in which the phenolic hydroxyl of the tyrosine at position 11 has been esterified to 8-dimethylaminonaphthalene-1-sulfonate (dansyl). This derivative fluoresces strongly in the visible with an emission maximun in dioxane of 530 nm, an emission lifetime of 16 ns, and a quantum yield of 0.8. Veatch et al. ((1975),J. Mol. Biol. 99, 75) have shown this 0-dansyltyrosine gamicidin C to be a fully active analogue of gramicidin A in artificial lipid bilayer membranes. We here utilize this derivative to further characterize the state of aggregation and rotational mobility of the four interconvertible conformational species formed by gramicidin in nonpolar organic solvents (Veatch et al. (1974), Biochemsitry 13, 5249; Veatch and Blout (1974), Biochemistry 13, 5257). Fluorescence energy transfer from the tryptophans of gramicidin A to the 0-dansyltyrosine of this derivatives supports the conclusion that all of these gramicidin isolated species are aggregates. Decay of fluorescence polarization anisotropy measurements yield a rotational correlation time of 1 ns for the 0-dansyltyrosine chromophore in ethanol in good agreement with the more detailed information previously obtained by 13C-nuclear magnetic resonance for the monomer in dimethyl sulfoxide (Fossel et al. (1974), Biochemistry 13, 5264). However, it is likely that the chromophore has much more rotational mobility than the rest of the gramicidin molecule in the aggregated comformational states.


RNA synthesis in Streptomyces antibioticus: in vitro effects of actinomycin and transcriptional inhibitors from 48-h cells. Two forms of DNA-dependent RNA polymerase have been partially purified (about 100-fold relative to the crude extract) from 48-h old cells of Streptomyces antibioticus. The two forms show different Mg2+ optima for the incorporation of [3H]UMP into RNA. Substances inhibiting transcription have been isolated by ammonium sulfate precipitation from one of the fractions produced during the polymerase purification. Actinomycin can be shown to inhibit RNA synthesis catalyzed by the S. antibioticus polymerases to a similar extent regardless of the template used. When S. antibioticus DNA is the template, actinomycin inhibits transcription by S. antibioticus polymerase to a degree that is significantly less than the observed actinomycin inhibition of synthesis catalyzed by Escherichia coli polymerase or by either S. antibioticus or E. coli polymerase with calf thymus DNA as the template. Using an assay previously developed, it was shown that the association constant for the binding of actinomycin to S. antibioticus DNA was increased by the presence of RNA polymerase in the binding mixture, while the association constant for the binding to calf thymus DNA was decreased by RNA polymerase. RNA synthesis in crude, cell-free extracts of 12-h old S. antibioticus cells (not producing actinomycin) is less refractory to actinomycin inhibition than synthesis catalyzed by extracts of 48-h old (actinomycin producing) cells, and both extracts catalyze appreciable RNA synthesis at actinomycin concentrations that completely inhibit RNA synthesis catalyzed by E. coli extracts.


On the association of reverse transcriptase with polynucleotide templates during catalysis. The association of avian myeloblastosis virus (AMV) DNA polymerase with polynucleotide templates during catalysis has been studied. During the course of polymerization, different template-primer complexes were added and the ability of the enzyme to switch from one polynucleotide template to another was determined. At 37 degrees C as well as at 4 degrees C, the polymerase is able to switch from certain template-primer complexes to others. For example, the addition of poly(A)-oligo(dT) during the course of synthesis with poly(C)-oligo(dG) results in the immediate cessation of dGMP polymerization and the start of dTMP polymerization without any lag. Early during the course of polymerization, the size of the product, as determined by alkaline sucrose gradient centrifugation, is, in part, a function of the ratio of the template-primer complex to the enzyme. These cumulative experiments indicate that catalysis on polynucleotide templates with avian myeloblastosis virus DNA polymerase under the conditions tested is not processive in a classical sense. Similar to cellular DNA polymerases the enzyme can shift from one template-primer to another. Using autoradiography after gel electrophoresis to estimate the product size, it can be calculated that the enzyme switches from one template to another within 0.25 min at 37 degrees C which corresponds to the incorporation of greater than 25 nucleotides. At 4 degrees C, switching can be calculated to occur in less than three nucleotide addition steps. Thus, with certain homopolymers, conditions can be found by which AMV DNA polymerase can switch from one template-primer complex to another, perhaps after each nucleotide addition step.


Observations on the pyridoxal 5'-phosphate inhibition of DNA polymerases. Pyridoxal 5'-phosphate at concentrations greater than 0.5 mM inhibits polymerization of deoxynucleoside triphosphate catalyzed by a variety of DNA polymerases. The requirement for a phosphate as well as aldehyde moiety of pyridoxal phosphate for inhibition to occur is clearly shown by the fact that neither pyridoxal nor pyridoxamine phosphate are effective inhibitors. Since the addition of nonenzyme protein or increasing the amount of template primer exerted no protective effect, there appears to be specific affinity between pyridoxal phosphate and polymerase protein. The deoxynucleoside triphosphates, however, could reverse the inhibition. The binding of pyridoxal 5'-phosphate to enzyme appears to be mediated through classical Schiff base formation between the pyridoxal phosphate and the free amino group(s) present at the active site of the polymerase protein. Kinetic studies indicate that inhibition by pyridoxal phosphate is competitive with respect to substrate deoxynucleoside triphosphate(s).


Cholinergic sites in skeletal muscle. I. Denervation effects. Cholinergic interactions in systems derived from rat skeletal mixed muscle are detailed. The isotherms of the binding of [125I]diiodo-alpha-bungarotoxin over the range of 10(-10)-10(-5) M toxin have been separated into a "nonspecific" component exclusive to the toxin and a "specific" component that binds both the toxin and d-tubocurarine. The "specific" component appears to reflect two independent sets of binding sites. One of the sets has an affinity constant on the order of 10(9) M-1. Following denervation, the number of sites in this high-affinity set begins to increase after 3 days, reaches a peak (28-fold higher than normal) on the 8th day, and begins to decline. Similar results are obtained when sensitivity of this set to an antibody derived from patients with myasthenia gravis is examined. This sensitivity is reflected by the inhibition of the alpha-bungarotoxin binding by the myasthenic IgG fraction. Following denervation, sensitivity first appears on day 3 progresses coincidentally with the increase in new sites in the set. The charcteristics of this set suggest that it represents the acetylcholine receptor and that the new sites appearing during the course of denervation are extrajunctional receptor sites. The interaction with the myasthenic IgG indicates an antigenic difference between junctional and extrajunctional receptors. The second set of specific binding sites has an affinity constant on the order of 10(5) M-1. The number of sites in this set increases only fivefold as a result of denervation. The increase also begins between days 2 and 3. The definition of this low affinity set of sites is not presently clear.


Effect of viomycin on dihydrostreptomycin binding to bacterial ribosomes. Viomycin, a peptide antibiotic, reduced the amounts of dihydrostreptomycin bound to ribosomes of Myobacterium smegmatis and Escherichia coli, although they have different modes of action. The [3H]dihydrostreptomycin binding to ribosomes could not exchanged with streptomycin or dihydrostreptomycin, but not with unrelated antibiotics, namely, kanamycin, neomycin, spectinomycin, capreomycin, tuberactinomycin-N, chloramphenicol and erythromycin. We suggest that there is a significant interaction between the binding sites of viomycin and streptomycin on ribosomes.


The chemical nature of osmium tetroxide fixation and staining of membranes by x-ray photoelectron spectroscopy. X-ray photoelectron spectroscopy was used to determine the oxidation states of osmium compounds present in erythrocyte ghost preparations and related systems treated with osmium tetroxide. Osmium tetroxide and cholesterol, codeposited at -100 degrees C, began to react at -70 degrees C, and Os(VI) was formed. Similarly, Os(VI) was detected for the known cholesterol-osmate ester prepared and purified chemically. However, osmium tetroxide applied in phosphate buffer (pH 7.2) gave rise to large proportions of Os(IV) and Os(III) species in addition to Os(VI) compounds. Egg phosphatidylcholine likewise produced a mixture of Os(VI), Os(IV), and Os(III), but dipalmitoyl phosphatidylcholine failed to give significant amounts of osmium containing products under identical conditions. Glutaraldehyde gave a mixture of compounds with the same osmium oxidation states when allowed to react with aqueous osmium tetroxide. Unfixed and glutaraldehyde-fixed erythrocyte ghosts also produced mixtures of Ss(VI), Os(IV) and Os(III) under conditions identical to those of normal tissue processing. Additionally, the mixture of adducts initially formed by treatment with osmium tetroxide was further reduced by dehydration of the tissue with ethanol, rpesulting in a final mixture which was 50-60% Os(III). The results support a scheme for the reaction os osmium tetroxide with tissues in which the initial reaction site is the double bonds of unsaturated lipids to form Os(VI) derivatives. Subsequent hydrolysis and further reduction yield complexes of Os(IV) and Os(III). A mixture of these three states is present in membrane specimens during microscopic observation. Os(VI) and Os(IV) could be present as osmate esters and osmium dioxide, respectively; Os(III) could be present as an oxo- or amino complex(es). The photoelectron spectrum of intact erythrocyte ghosts can be synthesized from the spectra of phospholipid and cholesterol only, suggesting the predominance of the reaction with lipids in the fixation process.


[Galactose transfer in the membranes of human milk fat globules (author's transl)]. Galactosyltransferase which catalyzes the transfer from UDP-galactose to either endogeneous glycoproteins, free N-acetylglucosamine or N-acetylglucosaminyl residues in the carbohydrate portion of glycoproteins, or to glucose when alpha-lactalbumin is added, occurs in human milk fat globule membranes. Various treatments (washing of membranes, freezing and thawing) did not affect this activity. In the presence of Triton X-100, the enzyme shows appreciable latency, This detergent was then used to solubilize the enzyme and to study its main characteristics. A competition and a heat stability experiment show that only one enzyme acts on two substrates (free N-acetylglucosamine or desialyzed and degalactosylated fetuin). UDP-galactose hydrolase activities were very low compared to those of the bovine milk fat globule membranes. Other characteristic enzymes of Golgi vesicles were found in human milk fat globules membranes. It is of interest to find out whether this is the result of contamination with cytoplasmic particles or whether it reflects the participation of Golgi vesicles in human milk fat globule secretion.


Immunochemical studies of proteins in epidermal cornified cells of human and newborn rat. 1. Proteins were extracted from cornified cells of newborn rats and human palm with 8 M urea containing 0.1 M beta-mercaptoethanol. Two fractions, rat FIIIa and human F5.5 were obtained by acid precipitation for further study. 2. Antibodies raised in rabbits by injection of rat FIIIa gave two precipitin lines by agarose diffusion against rat FIIIa, but only one line against human F5.5. One of the antigenic determinants of rat FIIIa was found to be a protein of approximately 66 000 daltons. The other seems to be formed with two polypeptides in the range of 60 000 and 66 000 daltons. The antigenic determinant of human F5.5 was a protein of approximately 64 000 daltons which immunologically cross-reacted only with the antiserum to a protein of 66 000 daltons in rat FIIIa. 3. The antisera also cross-reacted with proteins extracted from epidermis of guinea pig, hamster, hairless mouse, dog ear, dog snout and dog foot pad, but did not react with the epidermis of either rabbit immunized with rat FIIIa or non-treated normal rabbit. 4. Indirect immunofluorescence demonstrated a reaction of rabbit anti-rat FIIIa serum over cornified cells as well as over granular, spinous and basal cells of the epidermis of newborn rat and human.


Purification of alpha 2-macroglobulin with trypsin-like activity from pleural fluids. An alpha 2-macroglobulin with trypsin-like activity has been purified from pleural fluids of patients suffering from chronic pancreatitis. The isolation procedure includes ammonium sulphate precipitation, gel-filtration on Sephadex G-200 and DEAE-cellulose chromatography. It gives 46-fold purification of alpha 2-macroglobulin with a 13% recovery. Based on titration experiments with pancreatic inhibitor, the protein from three different patients contained 0.28, 0.46 and 0.80 mol of trypsin-like protease per mol of alpha 2-macroglobulin.


Manganese metabolism in cows and goats.

Manganese metabolism in cows and goats. When 54MnCl2 was incubated with fresh bovine or caprine serum for 20 h and the serum subjected to electrophoresis at pH 9.5, the 54Mn bound to transferrin and alpha2-macroglobulin in proportions which varied with the temperature of incubation and the temperature of electrophoresis. Between 0 and 37 degrees C, the higher the temperature of incubation the larger the proportion bound to transferrin and the lower the proportion bound to alpha2-macroglobulin. The temperature at which electrophoresis was performed had little effect on the proportion of 54Mn bound to transferrin, but increasing temperature reduced the proportion of 54Mn bound to alpha2-macroglobulin. Mn2+ did not bind to purified transferrin in vitro in the absence of an oxidising agent. In the presence of permanganate, Mn3+ was formed and chelated by transferrin at physiological pH. In fresh serum this oxidation step may be performed by ceruloplasmin or molecular oxygen. Mn2+ was bound reversibly to alpha2-macroglobulin but this protein played no part in the oxidation of divalent manganese and had no effect on the protein binding of trivalent manganese. Manganese in the divalent state, either free as Mn2+ or bound to alpha2-macroglobulin, is removed from blood plasma very efficiently by the liver. However, the manganic-transferrin complex normally found in circulation is not rapidly removed from plasma. The liver can remove large amounts of excess manganous manganese which it presumably excretes; the small essential fraction of the manganese absorbed is oxidised to the trivalent state and bound to transferrin.


Influence of serum and insulin on the accumulation of aminoisobutyrate by rat hepatoma cells. 1. Cultured rat hepatoma cells accumulate 2-aminoisobutyrate to high concentrations by a transport mechanism probably of the A type mediation. 2. Transport is enhanced by the presence of serum. When cells are deprived of serum the rate of transport declines over a period of hours; conversely addition of serum leads over a period of hours to increase in transport activity. In the presence of serum the apparent Km for aminoisobutyrate uptake is about 8 mM. In cells deprived of serum the Km is much higher. 3. Addition of insulin produces both an immediate increase in the rate of aminoisobutyrate uptake and a time-dependent rise. 4. The presence of alanine diminished aminoisobutyrate uptake in a concentration-dependent fashion. Competition is seen both in the presence and absence of serum but not when cells are incubated at 4 degrees C. 5. Preincubation with alanine for 1 h also diminishes aminoisobutyrate uptake when the alanine is removed. Cells take a period of several hours to recover from the depression of transport induced by alanine. 6. Transport of aminoisobutyrate rapidly declines in the presence of cycloheximide. Actinomycin had no effect for at least 8 h.


Preparation and characterization of two isozymes of choline acetyltransferase from squid head ganglia. II. Self-association, molecular weight determinations, and studies with inactivating antisera. The two isozymes of choline acetyltransferase (Acetyl-CoA:choline O-acetyltransferase, EC 2.3.1.6) from head ganglia of Loligo pealei have been examined by polyacrylamide gel electrophoresis, gel chromatography, and equilibrium sedimentation in the ultracentrifuge. Inactivating antisera, prepared to both native and dithiothreitol-treated isozymes 1 and 2 of squid choline acetyltransferase, were used to demonstrate the immunologic identity of isozymes 1 and 2. Each isozyme appeared to contain two non-identical catalytically active subunits, with molecular weights of approx. 37 000 and 56 000. A staining method was developed to visualize choline acetyltransferase activity in acrylamide gels. The method is based on the formation of a precipitate of manganese ferrocyanide at sites where free coenzyme A is released. By this method, and by analysis of gel slices, it was found that each of the isozymes can form aggregates of several different sizes. The formation of immune precipitates with the aggregates showed the identity of the multiple bands of enzyme protein resolved on disc gel electrophoresis. Isozyme 1 was most active as a small aggregate, whereas isozyme 2 was most active as a large aggregate. Both chromatography on Sephadex G-200 and isoelectric focusing yielded a number of active species with molecular weights ranging from 35 000 to 300 000. In addition, we demonstrated the dissociation of enzyme protein in the presence of 1.0 - 10(-2) M dithiothreitol, the formation of multiple precipitin bands by aged enzyme, and the identity of the different isoelectric fractions of each of the isozymes.


[RNA metabolism in chick embryo skin].

[RNA metabolism in chick embryo skin]. Explants from 7, 8, 9, 11, 13-day chick embryonic skin incorporating (3H) Uridine for different periods 1 hr, 3 or 4 hr and a chase with actinomycin) are studied with respect to free (F) or membrane bound (B) cytoplasmic polysomes and to RNA extracted from them. Polysome specific activity decreases at older stages but the amount of polysomes increases due to increased protein synthesis. At each stage B polysomes are less abundant but more radioactive than F polysomes. RNA extracted from each kind is analysed on sucrose gradients: one half of each fraction is precipitated by TCA to estimate total radioactivity, the other is retained on millipore at high salt concentration to estimate radioactivity in messenger-like RNAs due to their poly-A sequences. The pattern of the labelling of the different fractions of RNA changes with the length of incorporation, the stages of explants and the kind of polysomes (F or B); at 11-13 days the incorporation is slow, radioactivity is low and distributed among several peaks of poly-A RNA; at 7-8 days the incorportion is rapid, dispersed throughout the gradient; at 9 days, a midway stage, incorporation is particularly high into 12S and 24S fractions from B RNA. In the 5 studied stages the labelling of this 12S occurs early, remains for a longer time and cannot be chased. These observations suggest stability of the 12S RNA. Since, in 14-day chick embryos, feather keratin m RNA has been shown to sediment at 12S and although our experiments have been done with total skin because this differentiating tissue is the site of extensive interactions between dermis and epidermis, they suggest that this 12S RNA is the actual keratin m RNA and might be synthesised some days before the onset of keratin synthesis. Its template ability will be investigated at earlier stages.


Studies on amikhellin. II.-Inhibition of dna-polymerase from murine sarcoma leukemia virus. We have previously reported that amikhellin binds to double-stranded DNA by an intercalation process (1). We now report that this drug inhibits the DNA-polymerase from murine sarcoma leukemia virus. The extent of inhibition was found to vary with the nature of the primer-template used : maximum with poly(rA)n-oligo(dT)10 (nucleotide ratio 20:1), minimum with poly(rA)n-poly(dT)n and intermediate with native calf thymus DNA. Experiments performed with synthetic templates of the (rA)-(dT) type have led to the following conclusions as to the mechanism of inhibition: 1) Amikhellin acts at an early stage of the synthesis reaction because the drug is no longer inhibitory when a limited extension of the oligo(dT) primers has been allowed to occur. However, mere incubation of the enzyme with the template in the absence of dTTP is not sufficient to confer resistance to the drug. 2) Progression of enzyme molecules actively engaged in polymerization is stopped when they reach downstream duplex regions to which amikhellin is bound.


Alpha-foetoprotein and oestrogen metabolism. I. -Influence of alpha-foetoprotein on the metabolism of steroids by rat liver microsomes in vitro. Rat alpha-foetoprotein (AFP) was shown to inhibit the formation of water soluble metabolities of oestrone and oestradiol by incubation with microsomes from rat liver in the presence of NADPH. The results support the proposal that in young animals the low activity of enzymes responsible of oestradiol metabolism may be due in part to the presence of AFP and not only to the low level of these enzymes.


[Primary liver cancer and hepatitis B infection in Senegal. Comparison of cancer patients with 2 control groups]. A case/control study has been carried out to determine the prevalence of hepatitis B surface antigen (HBsAg) and antibody (anti-HBs) in patients with primary liver cancer (PLC), and age/sex matched hospital controls with cancers of other sites (OCC) and similarly matched controls without cancer (NCC). HBsAg was found in 61.2% of 165 cases of PLC, as compared to 11.7% of 154 OCC and 11.3% of 328 NCC. The frequency of HBsAg in PLC patients was significantly higher (72.2%) in those with detectable alpha-fetoprotein as compared to those without (40.3%).


Mesangial cell dysfunction detected by accumulation of aggregated protein in rats with streptozotocin induced diabetes. Diminished renal glomerular mesangial phagocytic function has been found in rats with streptozotocin induced diabetes. Similar impairment was produced by high dose cortisone and growth hormone, whereas oestrogen had a stimulant effect. The findings could be relevant to the understanding of human diabetic nephropathy.


[Detection of the base level of alpha-fetoprotein in the sera of donors by the electrophoresis-precipitation of polyacrylamide gel method]. Alpha-fetoprotein (AFP) was detected in the sera of adult healthy persons by the method of electrophoresis-precipitation in polyacrylamide gel; it was distinctly determined in about 50% of cases, and its concentration was not over 3ng/ml. AFP was determined all the 20 cases suffering from cirrhosis of the liver examined by the mentioned method.


[Changes in the chromatin structure of hepatocyte nuclei of rats trained to hypoxia]. Structure of chromatin in the nuclei of the isolated surviving hepatocytes and in the isolated nuclei of hepatocytes were studied by fluorochroming with acridine orange and by microfluorimetry of fluorescenc connected with the stain chromatin at 530 and 590 nm in intact rats and in the animals trained to hypoxia in a pressure chamber for 60 days. The nuclei of hepatocytes of intact rats were distributed by fluorescence at 530 nm into three classes with the intensity ratio of 1:2:4; as to the nuclei of hepatocytes of the rats trained to hypoxia - they formed a single class corresponding to the second class of control. In intact rats the ratio of the fluorescence intensity at 590 nm to such at 530 nm (alpha coefficient) formed normal distribution; in trained rats - a bimodal distribution with a shift of the maximum in the direction of reduction and increase of alpha in comparison with control. It is supposed that in hypoxia there is a repression of one and depression of other genes in the chromatine of the nuclei of the liver.


[Anti-arrhythmic activity of the beta2-adrenoblockader alpheprol]. Experiments were conducted on dogs, cats and rats in which various disturbances of the cardiac rhythm were induced. Beta2-adrenoblocker alpheprol proved to produce a marked antiarrhythmic action. The preparation arrested the auricular arrhythmias caused by electric stimulation of the auricles or by aconitine application, eliminated the auricular arrhythmias resulting from occlusion of a branch of the coronary artery or oubaine intoxication, prevented lethal auricular fibrillation in rats after the calcium chloride intoxication. It is supposed that along with the beta-adrenoreceptor block the antiarrhythmic effect of alpheprol was also caused by quinidine-like action of the preparation.


[Differences in the capacity of HLA sera to react with T- and B-lymphocyte populations]. A study was made of perculiarities attending the reaction of the HLA-sera with the T- and B-lymphocytes isolated from human blood. Lymphocytes were separated by removal of one of the cell subpopulation. T-lymphocytes were separated by the method of rosette-formation with sheep erythrocytes, with subsequent gradient density centrifugation. B-lymphocytes were separated similarly with the aid of rosette-formation with allogenous Rh-positive erythrocytes sensitized with Rh-sera with incomplete antibodies, and also sorption of B-lymphocytes on synthetic fiber. The cytotoxic activity of HLA-sera decreased after the removal of B-cells. But removal of T-lymphocytes was not accompanied by any reduction in the lymphocytotoxic activity. It is suggested that B-lymphocytes contained on their surface more HLA determinants than T-lymphocytes.


[Interaction of anti-Kb and anti-Dd effector lymphocytes with target cells of mouse mutant haplotypes]. The magnitude of the cytotoxic effect (CE) of immune lymphocytes R101 anti-Kb (C57BL/6) and R101 anti-Dd (B10.D2) upon target cells (TC) of mutants Kba (Hzl) and Kbd (M505), as well as Dda (M504) was considerably less than that on the TC of the original C57BL/6 and B10.D2 strains, respectively. A decrease of the CE on the H (zl) and M504 TC was more prominent than that on the M505 TC. "Monospecific" lymphocytes anti-Kb and anti-Dd were separated into two distinct subpopulations by absorption on the monolayers of macrophages from the mutant strains of mice. One of the subpopulations reacted with the TC of the original strain only, whereas the other one - with the TC of both the original and mutant strains.


[Immunochemical identification of human placental organ specific alpha2-globulin and its concentration in amniotic fluid]. An organospecific alpha2-globulin of human placenta was identified with the aid of immunochemical analysis. This antigen differed immunologically from the alpha2- and beta1-globulins of pregnancy, alpha-fetoprotein, placental lactogen, and chorionic gonadotropin. High level of the antigen revealed was found in the placental tissue and the amniotic fluid at the early terms of pregnancy, but its concentration sharply decreased by the time of labour.


[Antigenic structure of phytohemagglutinin-stimulated mouse lymphocytes using specific antisera]. The use of antiserum to the intact and the PHA-transformed lymphocytes showed that a new antigenic determinant (or determinants) appeared on the surface of lymphocytes after 68-hour PHA-stimulation. Intact lymphocytes or cells stimulated by the PHA for 2 hours only didn't carry this antigenic marker. At the same time the quantity (or density) of the antigenic markers present on the surface of the intact lymphocytes was lowered in the lymphocytes stimulated with the PHA for a long time.


5-Azacytidine (NSC 102816): a new drug for the treatment of myeloblastic leukemia. The pyrimidine analog, 5-azacytidine (NSC 102816), was administered by continuous intravenous infusion in Ringer's lactate in increasing doses to sets of patients with metastatic cancer to establish a dose sufficient to produce mild toxicity. Twenty-one patients (23 trials) were treated with doses of 50-200 mg/sq/m/day for 5 days every 2-4 wk. Nausea and vomiting were moderate and easily preventable. Doses of 100-200 mg/sq/m for 5 days every 14 days produced granulocytopenia, usually after two courses. Less toxicity was observed when courses were given every 21-28 days. Forty-five patients with previously treated and refractory acute myeloblastic leukemia were treated. The majority received doses of 150 mg/sq m for 5 days every 2 wk. Eleven (24%) complete remissions and four partial remissions were observed. The number of courses to achieve remission averaged three and required an average of 59 days. Nine patients with blastic crisis of chronic myeloblastic leukemia and four with refractory acute lymphoblastic leukemia failed to respond. 5-Azacytidine administered by continuous infusion is well tolerated and is an active compound in acute myeloblastic leukemia.


Exomphalos and gastroschisis: a 10-year review. Ninety-six cases of exomphalos or gastroschisis managed at The children's Hospital, Sheffield, during the period 1964-74 are analysed. There were 306 cases with minor lesions, 30 with major lesions, 27 with a ruptured exomphalos and only 3 with gastroschisis. The overall mortality rate was 46-9 percent. Survival could be closely correlated with the birth weight, the nature of the primary lesion, the presence or absence of liver within the sac and the presence and severity of associated congenital anomalies. A "scoring system" based on these criteria is proposed to predict the prognosis of an infant with an exomphalos.


Carcinoma of the periampullary region: a review of a personal series of 87 patients. Eighty-seven patients with carcinoma of the periampullary region treated by the author are reviewed. The clinical presentation and investigations are reported, and it is emphasized that the poor prognosis of this disease is due to the delay in presentation and diagnosis. Consequently, in most cases treatment can only be palliative, and the various forms of treatment are discussed.


Transfer RNA may be axonally transported during regeneration of goldfish optic nerves. If [3H]uridine is injected into the eyes of goldfish during optic nerve regeneration, then the return of fibers to the optic tectum is accompanied by the appearance of [3H]RNA in the tectum. The amount of [3H]RNA arriving in the tectum is consistently greater than in non-regenerating controls and reaches maximum levels (more than 10 times controls) 24 days after optic nerve crush. When [14C]uridine is injected subarachnoidally 1 day prior to sacrificing, the amount of [14C]RNA in the tectum is approximately doubled throughout the regeneration period. In order to characterize the radioactive tectal RNA in these experiments, we have crushed the optic nerves of 15 fish, and 18 days later injected [3H]uridine into both eyes. Five days later [14C]uridine was injected subarachnoidally and all fish were sacrificed a day later. RNA was extracted and fractionated in 2.0% polyacrylamide gels. The amounts of 3H- and 14C-labeled ribosomal as well as small molecular weight RNAs were increased during regeneration. Analysis of the area under the 28S, 18S and 4-7S RNA peaks indicated a small increase in 14C radioactivity in each peak (1.2, 1.5, and 1.5 times control, respectively). On the other hand, 3H radioactivity showed the greatest increase in the 4-7S fraction (8.0 times control) whereas large molecular weight ribosomal fractions were approximately 3 times control. Electrophoresis of the RNA on 10% polyacrylamide gels demonstrated that all of the small molecular weight RNA was confined to the 4S (tRNA) peak. These results suggest that when optic nerves of goldfish regenerate, they may enter the tectum carrying 4S (transfer) RNA.


Association of endodermal sinus tumour, testicular teratoma and alpha1-fetoprotein. An endodermal sinus tumour in the retroperitoneal region was associated with the presence of alpha1-fetoprotein (AFP) in the patient's serum. At autopsy a simple cystic teratoma of the right testicle was also found. The association of these two tumours has been reported before. The classification of these malignant germ-cell tumours and an understanding of their evolution may be aided by the discovery that AFP is often found in the patient's serum.


Anaplastic neuronal tumors of brain.

Anaplastic neuronal tumors of brain. A multicentric neuronal tumor of brain with unique morphologic features is described. It is compared with four other markedly anaplastic brain tumors containing neoplastic neurons. Demonstration of axon processes, which must be carefully distinguished from other similarly stained structures, is essential for the recognition of such tumors. Recent experience suggests that anaplastic neuronal tumors are more frequent than is generally realized. It is suggested that axon stains should be more widely employed in the investigation of atypical or highly pleomorphic gliomas.


Effects of bleomycin on human tongue carcinoma cells as revealed by electron microscopy. Five patients with tongue carcinoma were treated for 20 days with bleomycin, 15 mg/day, given by one-shot infusion through the superficial temporal artery. Specimens were taken 7, 14, and 20 days after the initial administration for light and electron microscope observations. The first alterations observed were the decrease of electron-dense chromatin clumps in nuclei and, simultaneously, the segregation of nucleolar components, i.e., amorphous protein component segregated from fibrillar and granular components. Subsequently, numerous nuclear bodies appeared in the nucleoplasm, which were observed for the first time in human tongue carcinoma cell nuclei in cases of bleomycin treatment. On the other hand, the cytoplasmic alterations were recognized by the decrease in number of mitochondria and by the appearance of numerous free ribosomes followed by the formation of tonofilaments. The cells enlarged because tonofilaments flocked together to make cancer cell nests, which were gradually together to make cancer cell nests, which were gradually transformed into a keratinic structure showing the so-called cancer pearl pattern. Finally, cancer cells were degenerated nearly to necrosis; there was no evidence of recurrence of cancer. Although the precise mechanism of the effect of bleomycin on nuclear activities of human tongue cancer cells remains unsettled because of the lack of cytochemical analysis, it is believed that bleomycin inhibits DNA synthesis and might also activate ribosomal RNA synthesis.


Comparison of in vivo translation rates and messenger RNA levels of alpha2U-globulin in rat liver and Morris hepatoma 5123D. The synthesis of the male rat hepatic protein alpha2U-globulin has been examined in Morris hepatoma 5123D and male host liver using pulse incorporation of labeled amino acids in vivo, followed by immunoprecipitation of the newly synthesized alpha2U-globulin from the soluble protein fraction of liver and hepatoma tissue. It was found that no alpha2U-globulin synthesizes alpha2U-globulin at a normal level (0.9 to 1.0% of total hepatic protein synthesis). A variety of liver-derived cell culture lines also did not have alpha2U-globulin synthesis. The level of the specific mRNA coding for alpha2U-globulin can be quantitated using in vitro translation of polyadenylate-containing RNA in a Krebs II ascites cell-free translational system, followed by immunoprecipitation of the alpha2U-globulin synthesized in vitro. Using this technique, it was found that host liver contained alpha2U-globulin mRNA at normal levels, whereas hepatoma tissue contained no detectable mRNA coding for this protein. Thus, alpha2U-globulin synthesis is deleted in the minimal-deviation hepatoma 5123D as a consequence of the inability of that tissue to produce functional mRNA coding for alpha2U-globulin. The implications for the regulation of gene expression in malignant cells are discussed.


Flow microfluorometric patterns of human bone marrow and tumor cells in response to cancer chemotherapy. A rapid propidium iodide staining method was used for analysis of single-cell suspensions of bone marrow and tumor biopsies by flow microfluorometry. With this technique, information on the proliferative status of target tissues can be obtained within 10 min of sample removal. DNA histograms and labeling index of sequential bone marrow biopsies from a patient with Stage IV diffuse lymphocytic leukemia and treated with 1-beta-D-arabinofuranosylcytosine infusion showed pronounced reduction in the percentage of cycling cells. In contrast, sequential tumor biopsies from a melanoma patient on methotrexate-citrovorum factor rescue therapy showed no changes. In sequential bone marrow biopsies of 3 patients on high-dose methotrexate-citrovorum factor rescue, initial accumulation of cells in G1-S (Day 1) was followed by a significant proliferative response (Days 4 to 7) and return to pretherapy values. In contrast, no recovery similar to that of the bone marrow was seen in tumor cells.


Single-strand scission and repair of DNA in mammalian cells by bleomycin. DNA single-strand breakage by bleomycin treatment of cultured mammalian cells was demonstrated by the method of alkaline elution. Elution patterns from treated L1210 cells indicated that part of the DNA was extensively broken while the remainder was affected to a lesser degree. This biphasic effect, which was less prominent in human fibroblasts, may reflect a selective sensitivity either of part of the cell population or of part of the DNA within individual cells. In both cell types, the DNA damage was at least partially repaired upon incubation of the cells after removal of drug. Bleomycin did not inhibit the rejoining of X-ray-induced single-strand breaks. The production and repair of DNA single-strand breaks after bleomycin treatment were the same in normal human and xeroderma pigmentosum fibroblasts, indicating that these events do not require the excision endonuclease that appears to be defective in these ultraviolet light-sensitive xeroderma cells.


Effect of pH on the bleomycin-induced DNA single-strand scission in L1210 cells and the relation to cell survival. The susceptibility of cultured L1210 cells to bleomycin was investigated as a function of pH of the medium, and was compared with DNA damage measured by alkaline elution. With increasing pH of the medium, both cytotoxicity and DNA damage increased. This was observed in the effects of bleomycin on cell proliferation, colony-forming ability, and DNA elutability. The reduction of colony-forming ability correlated with DNA single-strand scission, and this correlation was independent of pH.


The subcellular localization of calcium in vertebrate smooth muscle; calcium-containing and calcium-accumulating structures in muscle cells of mouse intestine. The intracellular localization of calcium by means of cytochemical techniques was studied in smooth muscle cells of mouse intestine. When the lead acetate method according to Carasso and Favard (1966) was used calcium was found in mitochondria and sarcoplasmic reticulum and occasionally between the myofilaments. The active ATP-dependent accumulation of calcium into cell structures was investigated by the oxalate method (Heumann and Zebe, 1967). After appropriate treatment the only structures of smooth muscle cells which contained calcium oxalate (identified by microprobe analysis) were elements of the sarcoplasmic reticulum. The results are discussed in relation to the role of calcium in the control of muscle activity during the contraction-relaxation cycle.


Enzymatic in vitro synthesis of globin genes. Full-length, single-stranded rabbit globin cDNA, synthesized by AMV reverse transcriptase, apparently contains a small double-stranded sequence (hairpin) at the 3' terminus. This cDNA can serve as template-primer for E. coli DNA polymerase I, which synthesizes a strand complementary to the cDNA and covalently bound to it. The loop connecting the two strands can be cut by S1 nuclease. Reassociation, hybridization, and restriction endonuclease studies, as well as electrophoretic analyses, indicate that the sequential actions of reverse transcriptase, DNA polymerase 1, and S1 nuclease generate full-length, double-stranded synthetic globin genes.


[Trichocellular leukemia].

[Trichocellular leukemia]. Basing on 13 cases of hair cell leukemia the gross morphology, histopathology and electron microscopy of this peculiar neoplasis is presented. The tricholeukocytes, as seen in Giemsa's preparation and especially in phase and electron microscope are pathognomic for establishing the diagnosis. In some cases the hairy cells may also be recognized in a HE stained tissue section. The staining method with Amidoblack 10 B may be useful for demonstrating hairy surface of the losely arranged neoplastic cells. Using a method with reduced Protargol and especially Timm's reaction after alcohol-sulfide fixation, a strong positivity could also be noticed. Exceptionaly the surface of free tricholeukocytes was found to be coated by proteinaceous precipitate resembling feature in Hoeppli's phenomen.


[Morphological findings in the liver tissue of Australia antigen carriers]. In twenty-four blood-donors were performed, after the average period of seventy-four days following the evidence of HB Ag in blood, the punction biopsies of the hepatic tissue. In microscopical examination one finding corresponded with the picture of a moderate, chronically persisting hepatitis, and in the twenty-first sample we found non-characteristic alterations, especially in the form of matrix activation of Kupffer's cells and isolated, dispersing resorption histiocytary nodules. Demonstrated in ten blood-donors was HB Ag in the plasma of hepatocytes.


A comparison of biochemical and biological properties of standard and defective lymphocytic choriomeningitis virus. Lymphocytic choriomeningitis (LCM) virus infection of the mouse is the best-studied model of persistent viral infection. In cell culture, persistent LCM virus infections are associated with the production of large quantities of defective interfering (DI) LCM virus. These defective interfering particles cannot replicate by themselves yet can interfere with the replication of the standard virus and prevent the cytolytic effect caused by the standard virus. It is important to determine the mechanism of interference and to establish whether the DI virus plays a role in vivo. Biological and biochemical properties of the standard and DI virus particles and also virus enzymes are compared. Antigenic analyses reveal that cells releasing only DI virus particles have less cell surface expression of viral antigens than cells releasing the standard virus. In the animal model, the DI virus is shown to have a protective effect against the pathogenesis of the LCM virus disease both in the mouse and in the rat.


Relationship between major histocompatibility antigens and disease. Possible associations with human arenavirus diseases. Histocompatibility antigens, virus infections, and disease are discussed relative to avenues of research in humans with arenavirus infections. The data implicating a relationship between histocompatibility complexes in man and animals and diseases of the central nervous system are reviewed. Histocompatibility antigens may share common antigenic determinants with viruses, act as receptor sites for attachment of viruses, and be altered by viruses. In addition, genes regulating immune responses to a variety of natural and synthetic antigens are linked, in many species, to the major histocompatibility complex. Since injury associated with virus infections may be largely due to the activity of the immune system, study of immune response genes may provide insight into understanding resistance to disease. Further, histoincompatibility reactions can activate latent viruses with resultant disease.


[Effect of two insecticides, lindane and DDVP (dichlorvos) on mouse adrenal glands]. Poisoning of non-pregnant female mice by sublethal doses of Dichlorvos and gamma-BHC (Lindane) induces prominent adrenal alterations. A significant weight increase of the glands is observed while Norepinephrine and Epinephrine content is depleted after Dichlorvos intoxication.


Studies on the immunosuppressive properties of a pregnancy-associated alpha-macroglobulin. A pregnancy-associated serum glycoprotein was shown to have an inhibitory effect on several in vitro methods of immunological assessment. This suppressing activity was evident at physiological concentrations and did not appear to be due to cytotoxicity. Transformation, induced by agents often regarded as preferential stimulators of T lymphocytes (concanavalin A, phytohaemagglutinin, allogeneic cells and tuberculin) was significantly depressed by the alpha-globulin. However, this phenomenon was much less evident when lipopolysaccharide or goat anti-human F(ab')2 serum was employed to selectively stimulate B-cells. The glycoprotein also blocked antigen-induced inhibition of leucocyte migration and caused a significant reduction in the number of lymphocytes binding sheep erythrocytes, in the spontaneous rosette test. It is proposed that the non-specific inhibitory activity of the alpha-macroglobulin depends upon some direct effect exerted on the lymphocyte itself and that it is levelled primarily at the cell-mediated immune response.


Immunological properties of Tg carbohydrates: enhancement of Tg immunoreaction by removal of sialic acid. Immunoreaction of desialylated and native thyroglobulin was compared in sera of patients with anti-thyroglobulin antibodies by radioimmunoassay with 125I-labelled thyroglobulin. Two human thyroglobulins were iodinated both in vivo and by lactoperoxidase. After desialylation enhancement of immunoreaction was observed in several sera ranging from marginal to more than 100%. The effect was not due to iodination since it was reproducible with desialylated thyroglobulin labelled in vivo. In one serum (B.P.) a marked enhancement was only seen with one thyroglobulin suggesting that desialylation may unmask isoantigens of thyroglobulin. Glycopeptides prepared from human thyroglobulin inhibited the immunoreaction between native thyroglobulin and autoantisera. The results indicate that sialic acid masks antigenic determinants in human thyroglobulin and that carbohydrates might be the determinants involved in the process of autoimmunization.


HLA antigens in pernicious anaemia.

HLA antigens in pernicious anaemia. The increased frequency of HLA-B7 alone and HLA-A3/B7 together, in the same patient, has been confirmed in pernicious anaemia. There is no increase prevalence of HLA-A3 alone. No association has been found between the presence of serum intrinsic factor type 1 antibody and HLA-A3 or HLA-B7.


The contribution of plasma protease inhibitors to antiplasmin activity in man. 1. Human plasma contains a variety of proteins that are capable of inhibiting plasmin activity. Whole plasma possesses 'rapid' and 'progressive' plasmin-neutralizing activity: this study assesses the contribution of individual protease inhibitors to this plasmin-neutralizing property of plasma. 2. Rapid and progressive antiplasmin activities of human plasma correlate with alpha2-macroglobulin and alpha1-antitrypsin concentrations respectively. 3. Fluctuations in the amounts of the other measured inhibitors (antithrombin III, Cl in activator and inter-alpha-tryspin inhibitor) did not influence the measured antiplasmin activity.


Congenital sensory neuropathy with anhidrosis: orthopedic complication and management. This report of 2 sisters with an extremely rare severe form of congenital sensory neuropathy with anhidrosis demonstrates the differential diagnosis, the orthopedic complications, and management. A review of the literature reveals only 7 previously reported cases.


Hepatitis-B: a review.

Hepatitis-B: a review. The recent literature on various aspects of hepatitis-B is reviewed with emphasis on the interrelationships of viral structure, antigenic components, and host immune response in acute, chronic, and asymptomatic carrier states of the infection. The mode of replication and mechanisms of transmission are discussed. Special attention is paid to potential non-parenteral routes of spread. The role of hepatitis-B in associated immune complex diseases and in hepatoma is outlined. A guide to the interpretation of serologic tests for hepatitis-B associated antigen and antibody patterns is presented in relation to the clinical stage and prognosis of the infection. Therapy, except in conceptual terms, is not covered but a summary of the current status of active and passive immunization is given. The unresolved question of the infectivity of carrier medical staff for their patient contacts, and the reverse, is discussed.


A model interdisciplinary diagnostic and treatment nursery. A model interdisciplinary diagnostic and treatment nursery program for developmentally disabled children is objectively described to enable documentation of effectiveness and replication. The major components of the program were the parent-professional partnership and the objective specification by the interdisciplinary team of problems, treatment plan, and treatment methods for each youngster. The concept of efficiency index is used to document and compare program effectiveness for four treatment cases, one of which is presented in detail. Reasons for program effectiveness and potential problem areas are discussed.


Intervention with premature human infants. Stimulation has been given to young infants to prevent the development disabilities often associated with prematurity. A review of these experimental programs reveals a lack of knowledge as to the ecology of the premature infant. This is indicated in the assumption that premature infants are sensorily deprived and by the arbitrary differences in the nature of stimulation techniques. Despite various problems in methodological and outcome assessment, stimulated infants tend to perform at higher levels than control-group infants on measures of sensorimotor and development. It is suggested that future research should be directed toward specifying the environment and processing capacities of the premature infant, and that intervention studies should make use of the 4-group design proposed by Solomon and Lessac (1968).


Labelling of DNA and differential sister chromatid staining after BrdU treatment in vivo. A method of labelling DNA in vivo with 5-bromodeoxyuridine (BrdU) is described. After 6 h permanent subcutaneous infusion of BrdU in rodents (adult Microtus agrestis, pregnant NMRI-mice), cell nuclei which have undergone DNA synthesis during the BrdU treatment can be differentiated from the nuclei of other cycle stages by means of their altered staining behaviour after Giemsa. 24 h after the BrdU treatment, mitoses from both bone marrow of the adult animals and tissues from the fetuses showed a differential sister chromatid staining. In male M. agrestis, sister chromatid exchanges were most frequently found in the euchromatic part of the X and in the constitutive heterochromatin of both sex chromosomes.


Meiosis of trisomy 21 in the human pachytene oocyte. Behaviour of the supernumerary chromosome, identification of chromomere sequence and numerous sub-bands. Association modalities of the three 21 chromosomes were studied during pachytene in three trisomy 21 fetuses whose chromosomal constitution was identified following amniocentesis. -- Three classes of images were observed: a trivalent, a trivalent presenting an important asynaptic region of the long arm, and a bivalent accompanied by a univalent. Such behaviour is analagous to that observed in all trisomic organisms. -- We have been able to establish the sequence of chromomeres, whose number varies from 9 to 14 according to the state of contraction in the 21 chromosome. Each band is thus subdivided into several sub-bands: at maximal elongation 2 sub-bands for band p11, 4 for q21 and 3 for q222. In addition, the interchromomeric clear bands q221 and q223 are also subdivided by the presence of a very small chromomere. In this way, the G-bands visible on mitotic metaphase chromosomes result from the compression together of several chromomeres whose individuality disappears as chromosomal condensation increases with progression of prophase.


Negative silver staining in A-T and satellite DNA-rich regions of human chromosomes. Human metaphase chromosomes were stained with silver following a pretreatment with a heated alkaline solution. The most conspicuous feature of the stained metaphases was the omission of silver staining in the secondary constrictions of chromosomes 1,9 and 16, and on the distal Yq. Our evidence indicates that the negative silver binding is due to the preferential removal or alteration of non-histone proteins associated with these regions. The cytochemical significance of these findings is discussed.


[Propafenon, a new anti-arrhythmic drug (author's transl)]. Propafenon, at an initial oral dose of 900 mg, proved an effective anti-arrhythmic drug in 34 patients with frequent ventricular and(or) supraventricular extrasystoles at rest. Long-term ECG monitoring of seven patients revealed that this effect lasted about eight hours. It can be maintained by an oral dose of 300 mg propafenon at eight-hourly intervals, as demonstrated in 21 further patients. Rapid anti-arrhythmic action was achieved in six acute cases with 1.0-1.5 mg propafenon per kg body weight by quick infusion, and lasted for one to one-and-a-half hours. In such cases the drug should, therefore, be given synchronously by the oral route. Side effects related to the gastrointestinal tract, sinus automatism as well as atrio-ventricular and intraventricular conduction, but in no case were very serious.


Visible and non-visible EEG changes demonstrated by spectral parameter analysis. A change of the heart rate in patients with complete AV-block and externally controllable cardiac pace-makers was earlier shown to influence cerebral function, as measured by psychological tests. To determine whether this influence manifests itself in the EEG, 9 patients aged 43-83 years were examined, all having complete AV-block and externally controllable pace-makers. Their EEGs were recorded at different heart rates between 40 and 100/min at rest and during a mental arithmatic test. The EEG record was evaluated visually and analysed by means of Spectral Parameter Analysis (SPA), introduced by Zetterberg (1969). The changes in the spectral parameters in relation to the heart rates were analysed statistically, In several cases significant changes appeared but the interindividual variations were large. Certain typical patterns could, however, be observed and their importance as a measure of cerebral function due to changes in the cerebral blood flow are discussed. These EEG changes revealed through SPA did not correspond to any visually observable changes in the EEG records. Differences in the spectral parameters between rest and test situations were analysed. Some of the differences found could easily be observed by visual inspection of the EEG. The advantage with SPA was that the changes could be described both qualitatively and quantitavely because SPA separates the spectral properties of the signals into components corresponding to well-known components of the EEG signal.


Motor and electrical signs of epileptiform activity induced by penicillin in the spinal cords of decapitate cats. Convulsive activity was induced in functionally decapitate cat preparations by topical and by systemic administration of toxic amounts of penicillin. The paroxysmal movement patterns and the electrographic signs of spinal seizure activity recorded from spinal ventral and dorsal roots and from the dorsal surface of the spinal cord are described. Paroxysms of interictal myoclonic twitching as well as tonic and clonic ictal seizures reminiscent of epileptiform convulsions of intact animals were seen in the absence of descending influences from the brain. Tonic seizures consisted of flexion--extension sequences; co-contraction of antagonistic muscles was the rule. Clonic activity consisted of rhythmic discharges at 4--6/sec, In dorsal roots, electrotonically conducted paroxysmal negative potential shifts as well as antidromically conducted trains of impulses were recorded. Ictal paroxysmal waves of the cord dorsum potential consisted of either biphasic positive--negative sequences or of purely negative waves. Diphenylhydantoin effectively controlled spinal seizures in the absence of a functioning cerebellum. Diphenylthiohydantoin changed the pattern of seizures, suppressing all ictal activity and greatly enhancing the amplitude and frequency of interictal bursts. Three different barbiturates suppressed seizure activity, but diazepam was ineffective, indicating that the site of its clinical anticonvulsant action may be supraspinal. Seizure activity, once induced, continued for up to 18 h. Intravenous administration of penicillinase abolished seizures indicating that their usual persistence is caused by the presence of the drug in the tissue, not by an irreversible biochemical lesion.


Functions of primary afferents and responses of extracellular K+ during spinal epileptiform seizures. Paroxysmal activity in ventral roots induced by penicillin in decapitate cat spinal cords is associated with waves of depolarization of primary afferent fiber terminals. These paroxysmal depolarizations can be detected as spontaneously occurring negative dorsal root potentials (DRPs) and are associated with antidromic discharge of nerve impulses in dorsal root fibers; they can also be detected by testing the excitability of afferent nerve terminals by focal stimulation. Negative DRPs evoked by afferent nerve volleys are altered in waveform but not in amplitude during seizures induced by penicillin, although they are blocked by the administration of picrotoxin. While blocking afferent-evoked DRPs, picrotoxin does not interfere with paroxysmal DRP'S, INDICATING DIFFERENCES IN THE GENERATION OF THE Two phenomena, which nevertheless have some link in common, for the paroxysmal waves occlude the evoked DRP. Such occlusion would appear as blockade, if DRPs were recorded by condenser-coupled amplifiers. In the presence of pentobarbital penicillin suppresses evoked DRPs, but under such circumstances seizure activity is not observed. Extracellular potassium activity within spinal gray matter transiently increases during seizure activity. Such increments of potassium activity are maximal in the ventral horns. This and several other observations suggest that in decapitate spinal cords systemically administered penicillin induces seizures which originate in the ventral gray matter. Accumulation of excess potassium may be the cause of paroxysmal depolarization of afferent nerve terminals. Excess potassium while not playing a principal role in initiating seizures, may influence the course of seizures by depolarizing afferent terminals. Such depolarization probably enhances tonic background release of transmitter substance, may modify the effect of synaptic input, and may favor synchronization of waves of neural excitability through extrasynaptic mechanisms.


Correlation between dominant EEG frequency, cerebral oxygen uptake and blood flow. In 32 chronic patients the EEG in temporal and occipito-parietal bipolar leads was analyzed with a manual method giving a period frequency index. The cerebral oxygen uptake (CMRO2) was calculated from the arteriovenous oxygen difference between samples from the internal carotid artery and the jugular bulb respectively, and from measurement of cerebral blood flow with the 133Xenon clearance method. A strong correlation (r = 0.78, P less than 0.001) was found between the EEG frequency indices and the CMRO2 of the hemisphere from which the EEG was recorded. Significant correlations between the EEG indices and gray matter cerebral blood flow were also demonstrated, while the correlation between the EEG indices and the blood flow of the white matter was weak. No correlation at all was found between the EEG indices and the relative weight of the gray matter. We conclude that the EEG carries information about the metabolic activity of the brain tissue. The EEG frequency also correlates with the blood flow of the gray matter of the brain when tissue anoxia has not uncoupled the normal function--flow relationship prevailing in the brain.


Contingent negative variation and the distraction--arousal hypothesis. Thirty-two normal volunteers were tested in three conditions: (1) a constant-foreperiod reaction-time situation consisting of a flash--tone--key-press sequence (control condition); (2) the addition of a short-term memory task consisting of four letters presented within the flash--tone interval with the requirement that they be repeated after key-press to tone (letters--recall); (3) the presentation of letters without short-term memory (letters--no recall). The task involving short-term memory of letters produced a significant reduction in amplitude of CNV for central (Cz) and parietal (Pz) recording sites. The association of CNV decrease and lengthened reaction time to tone was interpreted as a CNV distraction effect. The accompaniment of this distraction effect by elevated heart rate levels and increased frequency of eyeblinks was considered a distraction--arousal association and an important source of disruption in CNV development. These results were interpreted as support for the distraction--arousal hypothesis and appear to provide a sensitive complex of four measures for the evaluation of psychological processes, including the assessment of psychotropic drug effects. Eyeblink frequency in particular appears to be a sensitive indicator of distraction--arousal processes and a potentially useful measure of disturbed psychological functioning. The finding in control conditions of lower CNV amplitude in frontal than in central and posterior recording sites was viewed as a distraction effect due to efforts at eye movement control. The possibility was raised that frontal areas of the brain mediate sustained (tonic) distraction effects whereas centro-parietal regions mediate phasic distraction effects, at least when produced by stimuli of a lexical nature.


[Vsiual evoked potentials and excitability cycle during simulated diving at -610 m in oxygen-helium atmosphere (Physalie VI)]. Visual evoked potential (VEPs) and visual excitability cycles (VECs) were studied during compression in an oxygen--helium breathing mixture, up to a pressure of 62 ATA, equivalent to a depth of 610 m in sea water (Physalie VI). Records were taken at atmospheric pressure (control responses in confined helium--oxygen atmosphere) and at different depths during compression and decompression; recording was always performed at the same hours; subjects were lying with closed eyes. Isolated or double flashes were delivered every 1.2 sec, from a stroboscopic lamp located in the tank, at a maximum distance of 20 cm from the eyes. Responses were recorded with hook electrodes left in situ throughout the experiment (mid-vertex occipital derivation). VEP and VEC changes occurred in three stages: (1) Between surface and -435 m, components II-V of the VEP decreased in amplitude, while component VI increased. The after-discharge, and the PEV to the second of a pair of stimuli were facilitated. (2) Below 435 m, latencies and amplitudes of waves II-V increased, while those of wave VI decreased. After-discharge and response to the second stimulus were reduced. (3) During decompression up to -120 m, VEPs and VECs again changed, in a way similar to compression from 0 to -435 m. VEP and VEC changes, which are thus opposite during compression, may depend upon speed of compression, although the latter factor is probably not alone. Other factors, like pressure itself or the nature of the gas mixture may well intervene. It is likely that the observed modifications are the result of various factors interacting at various depths.


Dependence of reward contingent positive variation (RCPV) and cortical synchronization on visual input in the cat. Cats trained to press a lever for milk reward display high amplitude low frequency electrocorticographic post-reinforcement synchronization (PRS) associated with epicortical steady potential shift, termed Reward Contingent Positive Variation (RCPV), over the parieto-occipital cortex. Previously, it was found that the PRS-RCPV phenomenon depends on unpatterned light input devoid of any conceivable information or conditional property. Although training in the dark most likely eliminated the "novelty" factor and the resulting orienting reaction to "light off" as a cause of PRS-RCPV suppression, a possibility could not be excluded that the absence of light had a negatively reinforcing property preventing the emergence of PRS-RCPV. Hence, in the present study an experimental paradigm was used in which "light off" and/or "tone on" cues signalled the availability of reward, the "light off" periods extending through the consummatory response. Seven out of 9 cats, despite 2-4 months of daily training, and 85% correct timing of bar press performance, never showed a PRS-RCPV response (one animal showed only sporadic and poorly developed patterns.) However, when the light was turned on during consumption, in 85% of such tests a fully developed PRS-RCPV emerged in a stimulus-bound manner. One animal was an exception and showed PRS-RCPV both in the absence and presence of light. It was concluded that light input plays a crucial role in the emergence of the PRS-RCPV phenomenon and therefore also participates in the integration of gustatory input.


The effect of propranolol on normal physiologic tremor. It has been demonstrated that intravenous propranolol produces a 34-60% decrease in normal physiological tremor. The decrease, in response to small doses of the drug (less than 12 mug/kg), was linearly proportional to the baseline tremor size and occurred only following a 10 min delay. It is possible that the effect on tremor is due to the formation of a highly specific centrally acting metabolite of propranolol.


Effects of melatonin on the photosensitive epilepsy of the baboon, Papio papio. The acute pharmacological effects of melatonin (5-methoxy-N-acetyl-tryptamine) were evaluated on behaviour and electrographic signs in the photosensitive baboon (Papio papio), including visually evoked potentials VEPs). Doses of 5 and 10 mg/kg i.v. had little effect, contrasting with the enhancement of spontaneous paroxysmal activity and latency of clinical signs elicited by intermittent light stimulation (ILS) produced by the 20 and 50 mg/kg doses. Moreover, a modification of the electrographic pattern of discharge during ILS-induced myoclonic responses was observed. These observations indicate a slight decrease of light sensitivity after melatonin. Only frontal VEPs were modified, with increase in amplitude of their late components for all doses tested. These effects lasted for 2-4 h. The participation of serotoninergic mechanisms involved in melatonin effect and in this model of experimental epilepsy are discussed.


Reflex effects and postsynaptic membrane potential changes during epileptiform activity induced by penicillin in decapitate spinal cords. The administration of a convulsant dose of penicillin enhanced the transmission of monosynaptic reflexes in spinal cords in which reflex transmission was feeble before the drug treatment, but it had little effect in cords where monosynaptic reflexes were powerful to begin with. Post-tetanic potentiation was not altered by penicillin. Polysynaptic reflexes were invariably enhanced by convulsant amounts of penicillin. Postsynaptic ("direct") inhibition was not affected in the seizure-free intervals in spinal cords treated with penicillin, but it seemed to be suppressed during tonic seizures. The disability of reflex inhibition during ictal discharges may be due to presynaptic depolarization of inhibitory terminals. Recurrent inhibition was partially suppressed in spinal cords treated with penicillin. Neurons in the dorsal and intermediate gray matter were sometimes excited, sometimes inhibited, and sometimes unaffected by seizure activity of their segment. Motoneurons in the ventral horns invariably participated in the interictal and ictal activity. The timing of clonic seizure sequences coincided with bursts of Renshaw cell discharges. Action potential of abnormal amplitude and configuration were frequently observed in convulsing motoneurons. Paroxysmal depolarizing shifts (PDSs) of motoneurons were similar to those observed by other investigators in neurons in experimental epileptic foci of the cerebral cortex, except that spinal PDSs were not followed by hyperpolarizing waves.


[Effect of ablation of the motor cortex or the cerebellum on postural-kinetic coordination in the cat]. 1. A cat, restrained in a hammock, stands with its feet on supporting trays each furnished with strain gauges to measure the isometric upthrust. Placing reactions are elicited by contact of the left or right forelimb with a moving tray. The placing movement is preceded and accompanied by a postural adjustment characterized by an increase of the upthrust exerted by the opposite forelimb. 2. After ablation of the motor cortex on one side, the placing movement of the contralateral forelimb is abolished for the first few days after the operation and thereafter is delayed. However, the postural adjustment of the ipsilateral forelimb persists, with the same range of latencies as before the operation. 3. After unilateral motor cortical ablation, the contralateral forelimb still takes part in the postural adjustment associated with the placing movement of the other forelimb. 4. In two cats which had 2 years earlier undergone a total cerebellectomy, the placing movement was lacking or appeared with a long delay, whereas a postural adjustment of the other forelimb could be observed, although reduced. 5. The results indicate that the nervous structures responsible for the movement and for the associated postural adjustment are separate but partially linked.


[Action of ketamine of paroxysmal discharges provoked by air emboli in the baboon Papio papio]. Ketamine was administered to 18 baboons (Papio papio) to disrupt epileptic discharges elicited through intracarotid air injection. Discharge suppression was obtained in all animals but could be preceded by a phase of enhancement of the epileptic patterns, which rendered necessary the injection of a supplementary dose. The mechanisms of Ketamine action are discussed, in connection with data from the literature. Some of the latter suggest that its use in proved epileptics facilitates paroxystic patterns or even convulsions while others emphasize its antiepileptic action. Present data corroborate the ambivalent action of this drug. However, its dominant depressant effect obtained with increasing doses is somewhat favourable to its administration to epileptic patients.


[Sleep and respiration in the syndrome "apnea during sleep" in the child]. A sleep apnea syndrome has been diagnosed in eight children (age range 5-14). Before undertaking therapeutic trials, sleep and respiration were extensively studied. Sleep and respiration were again analyzed 3 months after tonsillectomy and adenoidectomy (6 cases) or tracheostomy with insertion of valve (2 cases). Sleep induced apneic apisodes in these children who had normal respiration during wakefulness. Three types of apnea (central, upper airway, and mixed) were recorded in each case. The minimum number of apneas recorded during a single night was 75; the maximum was 816. Polygraphic monitoring demonstrated greatly disturbed sleep. Sleep changes were quantitative as well as qualitative. REM sleep percent was decreased, but stages 3 and 4 NREM sleep were also impaired. A relationship between stages 3-4 NREM sleep and respiration was noted: stages 3-4 sleep disappeared when apneic episodes were numerous; no apnea was recorded during stage 4 sleep. Follow-up nocturnal recordings of two tracheostomized children with valve open, then closed, confirmed this "stage 4/no apnea" relationship. Apneas were also noted to induce marked sinus arrhythmia during sleep.