Control sites in the sequence at the beginning of T7 gene 1. The DNA sequence of the fragment Hind.30, 378 bases long, from the beginning of gene 1 of T7 is presented. It contains the C promoter, two in vitro transcriptional terminator sites and a sequence of 171 bases which probably codes for the N terminus of the T7 RNA polymerase. The sequence also codes for the RNase III cleavage site before gene 1. The overlaps with the transcriptional terminators, The RNA transcript of the sequence about the terminators can be arranged in a set of alternative double-stranded hairpin structures. It is suggested that conversion between these structures may have a role in termination; this may be influenced by interactions with ribosomes and RNase III. The region of the C promoter between genes 0.7 and 1 thus contains several sites which may be involved in the control of transcription and translation.


Construction of a recombinant bacterial plasmid containing DNA sequences for a mouse embryonic globin chain. Messenger RNAs for mouse embryonic globins were purified from yolk sac derived eyrthroid cells in mouse fetuses. Double stranded DNAs complementary to these messengers were synthesized and blunt end ligated to a EcoRI digested and DNA polymerase I repaired pBR322 plasmid. Of the ampicillin resistant transformants, one contained a plasmid with globin-specific cDNA. The inserted sequence is about 350 base pairs long. It contains one restriction site for EcoRI and one restriction site for HinfI about 170 and 80 base pairs from one end. The insert is not cleaved by HindIII, HindII, BamHI, PstI, SalI, AvaI, TaqI, HpaII, BglI. A mixture of purified messengers coding for alpha chains and for x, y and z embryonic chains was incubated with the recombinant plasmid and the hybridized messenger was translated in a mRNA depleted reticulocyte lysate protein synthesizing system. The product of translation was identified as a z chain by carboxymethylcellulose cromatography. The recombinant plasmid is named "pBR322-egz" after embryonic globin z.


Interactions of 4', 6-diamidine-2-phenylindole with synthetic polynucleotides. 4', 6-Diamidine-2-phenylindole forms fluorescent complexes with synthetic DNA duplexes containing AT, AU and IC base pairs; no fluorescent complexes were observed with duplexes containing GC base pairs or with duplexes containing a single AT base pair sandwiched between GC pairs. The binding site size is one molecule of dye per 3 base pairs. The intrinsic binding constants are higher for alternating sequence duplexes than for the corresponding homopolymer pairs. With the exception of the four-stranded helical poly rI which exhibits considerable fluorescence enhancement upon binding of the ligand, none of the single- or multi- stranded polyribonucleotides and ribo-deoxyribonucleotide hybrid structures form fluorescent complexes with the dye. Poly rI is the only RNA which forms a DNA B-like structure (Arnott et al. (1974) Biochem. J. 141, 537). The B conformation of the helix and the absence of guanine appear to be the major determinants of the specificity of the fluorescent binding mode of the dye. Nonfluorescent interactions of the dye with polynucleotides are nonspecific; UV absorption and circular dichroic spectra demonstrate binding to synthetic single- and double-stranded DNA and RNA analogs, including those containing GC base pairs.


Selective staining by 4', 6-diamidine-2-phenylindole of nanogram quantities of DNA in the presence of RNA on gels. 4', 6-Diamidine-2-phenylindole.2HCl (DAPI) forms fluorescent complexes with double-stranded (ds) DNA but not with ds RNA as shown by fluorescence titration. The widely used dye ethidium bromide (EB) forms fluorescent complexes with both types of nucleic acids. Also, in contrast to EB, DAPI forms much weaker fluorescent complexes with single-stranded DNA than with ds DNA. These observations were utilized to develop staining procedures for the selective visualization of ds DNA on gels. The use of DAPI in addition to EB for staining makes possible the localization of ds DNA and other species of nucleic acids on a single gel.


Translation of specific vaccinia virus RNAs purified as RNA-DNA hybrids on potassium iodide gradients. A procedure has been developed for purifying specific mRNAs by hybridization to fragments of DNA and isolation of the hybrids by potassium iodide equilibrium buoyant density centrifugation. The hybrids obtained are essentially free of unhybridized RNA as well as double-stranded RNA. Moreover, the RNA in the hybrids is undamaged and can be translated in vitro. Application of this procedure to mapping vaccinia virus genes is described. A total of 34 polypeptides have been assigned to three regions of the viral genome.


Specific inhibition of capped mRNA translation in vitro by m7G5'pppp5'G and m7G5'pppp5'm7G. A unique set of diguanosine cap analogues containing a 5'-5' tetraphosphate linkage instead of the normal triphosphate was synthesized by chemical methylation of G5'pppp5'G. Both 7-methylguanosine products, m7G5'pppp5'G and m7G5'pppp5'm7G, acted as potent inhibitors of capped brome mosaic virus (BMV) RNA translation in the homologous wheat germ protein synthesis system. Inhibition of in vitro protein synthesis required the presence of the 7-methyl group on guanosine and was specific for capped mRNA. In comparison with the partial cap analogue, m7GTP, the methylated diguanosine tetraphosphate structures were 25-50 fold more potent inhibitors of in vitro protein synthesis. Analysis of the in vitro translation products of the four species of BMV RNA showed a differential sensitivity to inhibition by m7G5'pppp5'm7G.


Heterogeneity of biologically active deadenylated protamine mRNA components isolated from rainbow trout testes. Poly(A)+ protamine mRNA's were isolated from rainbow trout testes and deadenylated by treatment with calf thymus RNase H. Four subcomponents of deadenylated PmRNA (PmRNA1-4) were purified by electrophoresis on a 6% polyacrylamide gel in 8 M urea. Translation of each PmRNA subcomponent in the wheat germ S-30 cell-free system showed that all subcomponents are biologically active but each codes for two or more protamine polypeptides suggesting molecular heterogeneity. However, the deadenylated mRNA's can be categorized into two groups based on the spectrum of protamines whose synthesis they stimulate.


Sequential Sepharose chromatographic isolation of polysomes and polysomal RNAs depleted in nuclear RNA from Xenopus. Analyses of sequence complexities, stage specific gene expression, and mRNA sequence divergence require polysomal RNA preparations relatively free of nuclear RNA contamination. Conventional procedures for the isolation of uncontaminated polysomal RNAs which rely on sucrose density centrifugations are laborious and unsuitable for large scale isolations. We describe here a method using sequential Sepharose chromatography for isolating polysomes and polysomal RNAs depleted for nuclear RNA. Polysomes and polysomal RNAs isolated from livers of Xenopus stimulated to produce vitellogenin were capable of directing protein synthesis in vitro and showed little evidence of degradation. The polysomal RNAs contained less than 0.5% of nuclear RNA.


Nuclease activity associated with the Ustilago maydis virus induced killer proteins. An in vitro nuclease activity was found to be associated with the purified killer proteins of Ustilago maydis. The proteins are effective against single stranded RNA, single and double stranded DNA. Endonucleolytic activity was confirmed by cleavage of circular molecules of 0x174 and PM2. Double stranded RNA did not appear to serve as a substrate.


The putative promoter of a Xenopus laevis ribosomal gene is reduplicated. With the aid of a novel poly-dA tailing-partial restriction technique and S1-protection mapping, the 5' terminal coding sequence for the 40S precursor ribosomal RNA of Xenopus laevis has been exactly identified. Since the promoter sequence for the 40S RNA should lie close to its 5' terminal coding sequence, we are able to conclude that the "Bam-Island" sequence reduplication (1) almost certainly represents a promoter reduplication.


5' and 3' terminal nucleotide sequences of the RNA genome segments of influenza virus. EXtensive nucleotide sequence analysis of the 5' and the 3' terminal of the RNA segments of the genome of fowl plague virus, an avian strain of influenza virus, confirms the presence of a common sequence at the 5' terminus of each segment and a common sequence at the 3' terminus of each segment. Between the ends of each individual segment there is a complementary sequence which may be important in the control of transcription and replication of the genome. In addition, the probable sites of initiation of translation of fowl plague virus mRNA are indicated along with the corresponding NH2-terminal amino acid sequences of the virus polypeptides.


Cloning of chicken embryo tRNA genes using single stranded nucleosomal DNA highly enriched for tRNA complementary sequences. DNA from chicken embryo nucleosome tetramers (about 760 base pairs in size) was enriched for tRNA genes by RPC-5 chromatography. The enriched DNA was hybridized with chicken embryo total tRNA and the hybridized DNA isolated utilizing a) avidinbiotin interaction, b) diazobenzyloxymethyl paper, and c) high temperature RPC-5 chromatography. The obtained single stranded DNA highly enriched for tRNA complementary sequences was hybridized with total DNA from nucleosome monomers (140--190 base pairs in size) and the excess of non hybridized monomer nucleosome DNA removed by Sepharose 4B chromatography. The hybrid molecules obtained were made fully double stranded by incubation with E. coli DNA polymerase I, DNA ligase, and exonuclease III. DNA was inserted into plasmid pBR322 by G-C joining procedure and the recombinant DNA used to transform the E. coli strain chi 1776. More than 70% of the transformants obtained hybridize to chicken embryo total tRNA.


Entrapment of plasmid DNA by liposomes and their interactions with plant protoplasts. Lecithin and lecithin/cholesterol liposomes formed in aqueous solutions of DNA entrap covalently closed circular, open circular and linear DNA molecules of size up to at least 13 kilobases. The sequestered DNA molecules are efficiently protected against exogenous deoxyribonuclease action although nicking and linearization of circular DNA can be observed. The size of these liposomes ranges from approximately 0.5 to 7.5 mu with an average of 2.5--4 mu. DNA filled liposomes strongly interact with plant protoplasts under conditions inducing protoplast fusion. Results suggest that sequestered plasmid DNA can be transferred to protoplast nuclei.


Evidence for a partial RNA transcript of the small circular component of kinetoplast DNA of Crithidia acanthocephali. The major component of kinetoplast DNA (kDNA) in the protozoan Crithidia acanthocephali is an association of approximately 27,000, 0.8 micrometers (1.58 x 10(6) dalton) circular molecules apparently held together in a particular structural configuration by topological interlocking. We have carried out hybridization experiments between kDNA samples containing one or the other of the two complementary (H and L) strands of purified 0.8 micrometers molecules derived from mechanically disrupted associations and RNA samples prepared either from whole C. acanthocephali cells or from a mitochondrion-enriched fraction. The results of experiments involving cesium sulfate buoyant density centrifugation indicate that whole cell RNA contains a component(s) complementary to all kDNA H strands, but none complementary to kDNA L strands. Similar results were obtained using mitochondrion-associated RNA. Digestion of RNA/DNA hybrids and suitable controls with the single-strand-specific nuclease S1 indicated that 10% of the kDNA H strand is involved in hybrid formation. Visualization of RNA/DNA hybrids stained with bacteriophage T4 gene 32 protein revealed that hybridation involves a single region of each kDNA H strand, equal to approximately 10% of the molecule length. These data suggest that at least 10% of the small circular component of kDNA of Crithidia acanthocephali is transcribed.


Molecular cloning of three major sequence species from Rainbow trout protamine mRNA. Double stranded cDNA molecules complementary to purified Rainbow trout protamine mRNA have been cloned in the bacterial plasmid pBR322. In order to circumvent the problems associated with a heterogeneous cDNA probe when identifying recombinants, a comparative hybridisation technique was used which can resolve between closely related cloned sequences. Using this technique, selected recombinants were shown to carry sequences corresponding to separate major fractions of protamine mRNA. Partial nucleotide sequences of the inserts in two clones confirms this conclusion.


The molecular electrostatic potential of the B-DNA helix. VI. The regions of the base pairs in poly (dG.dC) and poly (dA.dT). The evaluation of the electrostatic molecular potential at important nucleophilic sites of the purine and pyrimidine bases in poly (dG.dC) and poly (dA.dT) and of the evolution of the potential through the series free bases-nucleosides-nucleotides-single polynucleotide helices-double helices enables the interpretation of the evolution of the corresponding reactivity of the bases towards a series of electrophilic carcinogenic and mutagenic reactants.


Stimulation of oligonucleotide binding of estradiol receptor complexes by accessory proteins. During purification of E2R using oligo(dT)-cellulose chromatography, a receptor accessory factor (RAF) was identified in the cytosol of mouse kidney. This factor stimulates the binding of purified E2R to oligo(dT)-, oligo(dC)-, and oligo(dA)-cellulose as well as to DNA cellulose. It is a heat-stable, trypsin-resistant protein with an apparent molecular weight of between 10 and 30,000 daltons. Although structurally unrelated, similar stimulation of oligonucleotide binding was seen with calf thymus histones and, to a lesser extent, egg white lysozyme. Individual histones, especially H2a, H2B, and H3, also facilitate rebinding of purified E2R to oligo(dT)-cellulose, while H1 is less effective. Furthermore, histones stabilize the holoreceptor during sedimentation at 4 degrees and 12 degrees C. The N- and C-terminal half molecules of H2b were generated by cyanogen bromide-mediated cleavage and the N-terminal half was found to duplicate the effects of the parent molecule, both in binding and holoreceptor stabilization. These data suggest that the in vivo binding of E2R to DNA can be modulated by accessory proteins of cytosol and nuclear origin.


Atomic resolution analysis of a 2:1 complex of CpG and acridine orange. Cytidylyl-3', 5'-guanosine and acridine orange crystallize in a highly-ordered triclinic lattice which diffracts X-rays to 0.85 angstrom resolution. The crystal structure has been solved and refined to a residual factor of 9.5%. The two dinucleoside phosphate molecules form an antiparallel double helix with the acridine orange intercalated between them. The two base pairs of the double helical fragment have a twist angle of 10 degrees and it is found to have a C3' endo-(3', 5')-C2' endo mixed sugar puckering along the nucleotide backbone as has been observed for other simple intercalator complexes. Twenty-five water molecules have been located in the lattice together with a sodium ion. The intercalator double helical fragments form sheets which are held together by van der Waals interactions in one direction and hydrogen bonding interactions in the other. The crystal lattice contains aqueous channels in which sixteen water molecules are hydrogen bonded to the nucleotide, none to the intercalator, five water molecules are coordinated about the sodium ion and four water molecules bind solely to other water molecules. The bases in the base pairs have a dihedral angle of 7 to 8 degrees between them.


Computer programs for analysis of nucleic acid hybridization, thermal denaturation, and gel electrophoresis data. Computer programs for the analysis of data from techniques frequently used in nucleic acids research are described. In addition to calculating non-linear, least-squares solutions to equations describing these systems, the programs allow for data editing, normalization, plotting and storage, and are flexible and simple to use. Typical applications of the programs are described.


Detection of labelled RNA species by contact hybridization. An improved contact hybridization technique for the analysis of labelled RNA species is presented. The method combines high sensitivity of detection with the high resolution of polyacrylamide gel electrophoresis and should be especially useful for the characterization of transient RNA precursor molecules. Its application to gene mapping is illustrated.


Absence of cytosine methylation at C-C-G-G and G-C-G-C sites in the rDNA coding regions and intervening sequences of Drosophila and the rDNA of other insects. Cytosine residues in C-G dinucleotides are frequently methylated in eukaryote DNA. In DNA of the dinoflagellate C. cohnii, the sequence C-MeC-G-G apparently renders Hpa II (C-C-G-G) incapable of digesting whole cell DNA in general, and rDNA in particular. Msp I, which also recognizes C-C-G-G but cleaves irrespective of methylation, degrades C. cohnii DNA and produces rDNA segments of 10.2 to 1.4 kb. We have applied this Hpa II/Msp I test to unfractionated DNA, and to rDNA and the rDNA intervening sequence of Drosophila virilis embryos and adults. There is no evidence of C-MeC-G-G sequences in either developmental stage of this species. Absence of G-MeC-G-C from coding and intervening sequences of rDNA was shown in comparisons of Hha I (G-C-G-C) cleavage patterns of unfractionated DNA and cloned (unmodified) segments of rDNA. Comparisons of Hpa II and Msp I cleavage products of DNA from the house fly, the flesh fly and a bumblebee also revealed no internal cytosine methylation in the sequence C-C-G-G. Because amounts of McC in C-G dinucleotides vary greatly among species, from apparent nonexistence to substantial proportions, no inference may yet be drawn about the role of such base modifications in DNA.


A regulatory sequence near the 3' end of sea urchin histone genes. The 3' flanking sequences of all five histone genes have been sequenced in the histone DNA clone h19 of the sea urchin Psammechinus miliaris. A large (23 bp) and a small (10 bp) conserved sequence was found by sequence comparison, some 29-40 bp downstream from the termination codon. 12 bases of the larger homology block show a dyad symmetry. The available sequences of clone h22 of the same species and those of the histone clones pSp2 and pSp17 of Strongylocentrotus purpuratus, another sea urchin species, fit well into this comparison. Two types of sequences are involved in the dyad symmetry; one is H1, H3 and H4 specific, the other is H2A and H2B specific. If these conserved sequences are transcribed, a hairpin loop could form in the RNA molecules. This secondary structure might serve as a recognition signal for a regulatory protein.


Preparation of triple-block DNA polymers using recombinant DNA techniques. The construction of several recombinant plasmid derivatives containing novel triple-block DNA sequence insertions is described. The protocol for these constructions involves synthesis of a heterogenous mixture of block oligomer duplexes, : formula: (see text), using pancreatic deoxyribonuclease and terminal transferase. The synthetic duplexes were mixed with linearized and dG-tailed vectors and the DNA mixture used to transform E. coli. Triple-block sequences of the type dGidAjdCk.dGkdTjdCi, characterized by DNA sequencing, were inserted into the Bam HI site of pBR322 and next to the lac wild-type and UV5 promoter regions in pRW26 and pRW28. Similarly, sequences were inserted into the Sma I site of pACYC189 and could be excised by cleavage with Sma I since the procudure regenerates the recognition site. The approach provides a technique for the synthesis of a large family of defined sequence triple-block polymers in essentially unlimited amounts. Although these inserts contain sequences which have the potential for forming stable hairpin structures, the recombinant plasmids are stable and appear to replicate normally.


Fine structure melting of viroids as studied by kinetic methods. The conformational transitions of five viroid species were studied by melting analysis and by fast and slow temperature jump techniques. Experiments with the fast temperature jump technique had to be carried out in 10 mM Na-cacodylate, 0.1 M NaCl, 4 M urea, 1 mM EDTA, pH 6.8. In addition to the highly cooperative main transition (Tm between 46.5 and 49 degrees C for different viroid species [1]) all viroids show at higher temperatures an intermediate transition (Tm approximately equal to 57 degrees C) and a high temperature transition (Tm approximately equal to 68 degrees C). The maximum amplitudes of these transitions amount only to about 1% of that of the main transition. The main transition represents a net dissociation of 78 to 94 base pairs depending on the viroid species. The intermediate transition corresponds to the dissociation of two hairpins with 5-10 base pairs each, and 10-20 nucleotides in the loops. The high temperature transition corresponds to a hairpin of 9 G:C pairs and 1 A:U pair and more than 40 bases in the loop. It is shown that these stable hairpins are not part of the native structure but are newly formed during the main transition. Their formation is responsible for the extraordinary cooperativity observed in the main transition. Hairpins can be correlated to defined sequences of PSTV. Based on these studies, on the sequence of PSTV [2], and on a theoretical treatment [3] a detailed description of the whole mechanism of PSTV denaturation is given.


The synthesis and processing of a nuclear RNA precursor to rat pregrowth hormone messenger RNA. A recombinant DNA plasmid, pBR322-GH1, which contains about 80% of the sequences of rat pregrowth hormone (pGH) mRNA, allowed an analysis of nuclear RNA from GH3 cells for possible precursors of cytoplasmic pGH mRNA. A single 20-22S RNA SPECIES ABOUT 2-3 TIMes larger than pGH mRNA was detected in nuclear RNA from GH3 cells labeled for 5 min. with 3H-uridine. After longer label times a 12S RNA indistinguishable in size from cytoplasmic  12S pGH mRNA became the predominant labeled RNA complementary to the plasmid pBR322-GH1. Both of these nuclear RNA species contained poly (A). Kinetic analysis of the labeling of nuclear and cytoplasmic pGH mRNA sequences showed that the 20S and 12S nuclear RNA molecules were labeled before significant labeling of cytoplasmic pGH mRNA was detected, and also indicated that there is complete conservation of nuclear pGH mRNA sequences in the production of cytoplasmic pGH mRNA. These results indicate that cytoplasmic pGH mRNA is generated by nuclear processing of a larger nuclear RNA molecule.


Molecular recombination and the repair of DNA double-strand breaks in CHO cells. Molecular recombination and the repair of DNA double-strand breaks (DSB) have been examined in the G-0 and S phase of the cell cycle using a temperature-sensitive CHO cell line to test i) if there are cell cycle restrictions on the repair of DSB's' ii) the extent to which molecular recombination can be induced between either sister chromatids or homologous chromosomes and iii) whether repair of DSB's involves recombination (3). Mitomycin C (1-2 micrograms/ml) or ionizing radiation (50 krad) followed by incubation resulted in molecular recombination (hybrid DNA) in S phase cells. Approximately 0.03 to 0.10% of the molecules (number average molecular weight: 5.6 x 10(6) Daltons after shearing) had hybrid regions for more than 75% of their length. However, no recombination was detected in G-0 cells. Since the repair of DSB was observed in both stages with more than 50% of the breaks repaired in 5 hours, it appears that DSB repair in G-0 cells does not involve recombination between homologous chromosomes. The possibility is not excluded that repair in G-0 cells involves only small regions (less than 4 x 10(6) Daltons).


Purification of histone messenger ribonucleoprotein particles from HeLa cell S-phase polysomes. Characterization of associated proteins. We have purified HeLa histone mRNA from polysomes of S-phase cells which had been synchronized by hydroxyurea treatment. This mRNA was shown to direct the in vitro synthesis of all five histones which amount to at least 90-95% of its total translational activity. Polysomal histone mRNP was also purified and identified by cell-free translation and hybridization to a clone of histone DNA from E. esculentus. The protein moiety of this mRNP contained three prominent species of molecular weight 86,000, 73,000 and 53,000 daltons. The presence of the 73,000 species previously assessed to be bound to poly(A) is discussed in view of the fact that histone mRNA does not contain a pail. As globin mRNA, histone mRNA as well as histone mRNP were translated with equal efficiency in cell-free extracts from either S-phase or hydroxyurea blocked HeLa cells.


Interaction of a limited set of proteins with different mRNAs and protection of 5'-caps against pyrophosphatase digestion in initiation complexes. A variety of 5'-3H-methyl-labeled, oxidized viral mRNAs were used as probes for detecting in wheat germ initiation complexes proteins that interact with, and can be cross-linked to, the 5'-cap structure. A limited and reproducible set of specific proteins was obtained with the different mRNAs. The binding of these proteins to the 5'-end of mRNA apparently results in protection against nucleotide pyrophosphatase digestion of the cap even in initiation complexes in which the 5'-end is susceptible to pancreatic RNase digestion. Cross-linked proteins from mammalian initiation complexes comigrated with several of the subunits of similarly treated eIF-3. A model for cap binding protein interaction with mRNA cap during initiation of translation is suggested.


Radioimmunoassays for the modified nucleosides N[9-(beta-D-ribofuranosyl)purin-6-ylcarbamoyl]-L-threonine and 2-methylthioadenosine. Radioimmunoassays were established for the modified nucleosides N-[9-(beta-D-ribofuranosyl)purin-6-ylcarbamoyl]-L-threonine, t6A, and 2-methylthioadenosine, ms2A. The assays depended on the production of antisera specific for t6A and ms2A that have not been previously reported. The nitrocellulose membrane filtration and saturated ammonium sulfate RIA techniques were compared for efficiency. Various radioactive antigens were employed to establish which type of antigen would give the best binding. The tritium post-labeling procedure of Randerath and Randerath was used to obtain labeled nucleosides of high enough specific activity to be useful for RIAs when the labeled nucleoside was not available commerically. The specificity of the antibodies toward nucleosides and purified tRNAs is reported. Although the titer of the t6A antiserum was low, the specificity was very sharp. An interesting finding was that threonine, a major structural component of the side-chain modification of t6A, was completely infective as an inhibitor.


8-Phosphorus substituted isosteres of purine and deazapurines. Synthesis of 8-phosphorus substituted isosteres of purine [pyrimidino (4,5-d)-1,3,2-diazaphosphole], 1-deazapurine [pyridino (2,3-d)-1,3,2-diazaphosphole] and 3-deazapurine [pyridino (4,5-d)-1,3,2-diazaphosphole] has been achieved by the reaction of equimolar amounts of triphenylphosphite and 4,5-diaminopyrimidine, 2,3-diaminopyridine and 3,4-diaminopyridine, respectively. These compounds hydrolyzed (cleavage of the phosphorus-nitrogen bounds) in aqueous solutions to provide the corresponding diaminopyrimidine or diaminopyridines. These three new basic ring systems constitute the first reported synthesis of purines in which ring carbon atom is substituted with a phosphorus atom. 8-Phosphorus substituted purine at a concentration of 4 X 10(-4)M caused a 50% inhibition in the growth of leukemia L1210 cells in culture. The biochemical rationale for the synthesis of these compounds is discussed.


Nonrepetitive DNA transcription in normal and regenerating rat liver. Initial RNA excess hybridization experiments employing total cell RNA and the complete complement of nonrepetitive DNA sequences showed no differences between normal and regenerating rat liver. However, when the DNA from the RNA-DNA hybrids was isolated and then reacted with homologous and heterologous RNAs the sensitivity of the assay was sufficiently improved to reveal that some of the nonrepetitive DNA transcrips present in normal liver are missing at 24 h and 48 h after a 70% partial hepatectomy. Additional experiments showed that while some of the missing sequences were common to both stages of regeneration, some were also different. The data thus suggest both quantitative and qualitative changes during liver regeneration in the population of RNA molecules transcribed from nonrepetitive DNA.


Thionein gene expression in Cd++-variants of the CHO cell: correlation of thionein synthesis rates with translatable mRNA levels during induction, deinduction, and superinduction. The relationship of thionein synthesis rates to translatable cytoplasmic thionein mRNA levels was investigated for the first time in a cultured cell system. Thionein synthesis was induced in Cdr, a cadmium-resistant variant of CHO, by exposure to 2 microM CdCl2. Following a short (1.5 hr) lag, thionein synthesis increases to a rate that is at least 30 times the uninduced rate 7-8 hr after addition of Cd++. This increase is blocked by the coincident addition of a actinomycin D. Cytoplasmic thionein mRNA levels, measured by translation in a modified wheat germ system, increase rapidly following induction to values approximately 25 times uninduced levels within 6-8 hr. The increase in thionein mRNA precede proportionate increases in thionein synthesis by 0.5-1.0 hr. Continued exposure to Cd++ results in a decreased thionein synthesis rate after 8 hr. By 30 hr, the rate is one-half that seen 6-8 hr after induction. Removal of Cd++ after 8 hr results in a rapid decrease in thionein synthesis (t 1/2 approximately 4 hr). Both decreases are inhibited by the addition of actinomycin. In all instances--induction, deinduction, and actinomycin-mediated "super-induction"--translatable thionein mRNA levels and thionein synthesis rates increase, decrease, or are maintained coordinately. The results suggest that thionein synthesis in Cdr is controlled primarily by the level of translatable cytoplasmic thionein mRNA.


The non-histone proteins of the rat liver nucleus and their distribution amongst chromatin fractions as produced by nuclease digestion. The search for proteins involved in maintaining higher order chromatin structures has led to a systematic examination of the non-histone proteins (NHP) of rat liver nuclei in the context of nuclease digestion studies. 40-45% of the 3H-tryptophan labelled NHP originally present could be removed by extensive washing in a "physiological" buffer, incubation at 37 degrees C with or without nuclease and a further wash step. Nuclei at this stage had a remarkably constant NHP content (ca. 0.73 micrograms/micrograms DNA), independent of the degree of digestion with micrococcal nuclease or HaeIII. The solubilized chromatin produced by limited digestion with either nuclease contained 0.3-0.5 microgram NHP/microgram DNA, this value falling to ca. 0.16 after more extensive cleavage. Insoluble chromatin fractions were between 2-fold (very limited digestion) and 16-fold (extensive digestion) richer in NHP than the corresponding soluble fractions. Gel electrophoresis revealed about 12 NHP bands in soluble fractions, the most prominent of M.Wt. 41.400, while the insoluble material had at least 50 components. These properties were independent of whether lysis of nuclei occurred in 0.2 or 50 mM ionic strength. The large disparity in NHP content between complementary soluble and insoluble chromatin fractions is considered in terms of chromatin organization in vivo and the possible role of NHP migration.


A partial characterization of DNA fragments protected from nuclease degradation in histone depleted metaphase chromosomes of the Chinese hamster. A small proportion (0.1-0.5%) of the total DNA content of native Chinese hamster metaphase chromosomes is protected from nucleolytic degradation following the removal of histones by extraction with either 0.2 N HCl or 2 M NaCl, and remains attached to the nonhistone protein core. Acid extraction followed by DNase I digestion leads to small fragments of 10-30 bases. Salt extraction followed by micrococcal nuclease digestion gives approx. 140 b.p. fragments which are undistinguishable in size from nucleosome core DNA fragments. Furthermore, DNase I treatment of salt extracted chromosomes gives DNA fragments containing single strands which are multiples of 10 bases in length, again characteristic of the nucleosome structure. Reassociation kinetics using the 32P-labelled 140 b.p. fragments as probes suggests they are enriched for rapidly reassociating sequences.


Extrachromosomal circular ribosomal DNA in the yeast Saccharomyces carlsbergensis. Purified ribosomal DNA from Saccharomyces carlsbergensis contains a small proportion of circular DNA molecules with a contour length of 3 micron or integral multiples thereof. Hybridization of yeast ribosomal DNA with 26 S rRNA, using the R-loop technique, reveals that these circular molecules contain sequences complementary to yeast ribosomal RNA. We suggest that these extrachromosomal rRNA genes may be intermediates in the amplification of rRNA genes in yeast.


On the de novo formation of compact oligonucleosomes at high ionic strength. Evidence for nucleosomal sliding in high salt. Histone H 1-depleted chromatin made from acid extracted, intact nuclei was exposed to various ionic strengths. NaCl concentrations above 0.3 M sufficed to generate novel oligonucleosomes formerly characterized as "compact oligomers" and "spacerless dinucleosomes". Such particles could not be identified within H 1-depleted nuclei or chromatin at low ionic strengths. Their formation, proceeding within days at 0 degrees C, was accelerated by increasing ionic strengths. The data was discussed in terms of a salt-induced motion of nucleosomal core particles along the DNA to form compact oligomers.


No more than seven interruptions in the ovalbumin gene: comparison of genomic and double-stranded cDNA sequences. We have determined the sequence of ovalbumin RNA (ov-mRNA) using a double-stranded cDNA (dscDNA) plasmid. We have also determined the sequence of the previously characterized exonic regions of the chicken ovalbumin gene. The comparison of these various sequences has shown that there are no additional interruptions in the mRNA-coding sequences above those 7 already characterized. There is only one single base discrepancy between the two mRNA sequences determined using the dscDNA or the genomic clones. This demonstrates the accuracy and reproducibility of the cloning and sequencing techniques. The ovalbumin mRNA sequence was found to be 1872 nucleotides in length, 13 nucleotides larger than the previous value reported by McReynolds et al. [Nature 273, 723-728 (1978)].


Localization on the viral genome and nucleotide sequence of the gene coding for the two major polypeptides of the hepatitis B surface antigen (HBs Ag). The structural gene coding for both polypeptides I and II which are the two major polypeptides of the Hepatitis B surface antigen, is found to be localized on the viral genome. This gene, referred to as gene S, is located in the partially single stranded region. It maps between positions 73.6 and 95.1% of the genome length. It is composed of 678 nucleotides, which correspond to a theoretical polypeptide of 25,422 molecular weight.


The distribution of histone H1 subfractions in chromatin subunits. Rat liver chromatin was digested with micrococcal nuclease to various extents and fractionated into nucleosomes, di and trimers of nucleosomes on an isokinetic sucrose gradient. In conditions under which degradation of linker DNA within the particles was limited, the electrophoretic analysis of the histone content showed that the overall content of H1 histone increased from nucleosomes to higher order oligomers. Moreover, the histone H1 subfractions were found unevenly distributed among the chromatin subunits, one of them, H1--3 showing most variation. A more regular distribution of these subfractions was found in subunits obtained from a more extended digestion level of chromatin. It is suggested that the H1 subfractions differ in the protection they confer upon DNA.


The nature of the interaction of nucleosomes with a eukaryotic RNA polymerase II. The integrity and stability of nucleosomes under transcription assay conditions has been found to depend on concentration and ionic environment. Rifamycin AF/013, a commonly used inhibitor of initiation, is particularly effective in destabilisation of nucleosomes. Intact nucleosomes are refractory to transcription by wheat RNA polymerase II, the histone core preventing initiation. Template titration suggests that the polymerase can, however, bind to nucleosomes, and a 15--16S complex has been observed on sucrose gradients. DNase I digestion of polymerase-nucleosome incubations indicates that whilst histone is still present in the complex, the nucleosome conformation is altered resulting in enhanced nucleolysis at sites near the DNA centre but reduced overall kinetics of digestion.


Nucleotide sequence of a highly repetitive component of rat DNA. A highly repetitive component of rat DNA which could not yet be enriched by density gradient centrifugation was isolated with the help of the restriction nuclease Sau3AI. This nuclease converted the bulk of the DNA to small fragments and left a repetitive DNA component as large fragments which were subsequently purified by gel filtration and electrophoresis. This DNA component which was termed rat satellite DNA I is composed of tandemly repeated 370 bp blocks. According to sequence analysis the 370 bp repeats consist of alternating 92 and 93 bp units with homologous but not identical sequences. Methylation of CpG residues was correlated to the rate of cleavage by restriction nucleases. Significant homologies exist between the sequences of rat satellite DNA I and satellite DNAs of several other organisms. The divergence of the sequence of rat satellite DNA I was discussed with respect to evolutionary considerations.


Conformation of DNA in chromatin core particles containing poly(dAdT)-poly(dAdT) studied by 31 P NMR spectroscopy. We have prepared semi-synthetic chromatin core particles from a complex of chicken erythrocyte inner histones (H2A, H2B, H3 and H4) with double-stranded poly(dAdT).poly(dAdT) and studied the conformation of the phosphodiester backbone using 31P NMR at 109.3 MHz. At 20 degrees C, the core particle spectrum is fit well by a single Lorenzian distribution with a line width of 110 Hz. This signal is significantly broader than that for the 145 base pair poly(dAdT).poly(dAdT) alone; the latter consists of two resonances, approximately equal in intensity, with average line width 41 Hz. Major changes in the spectrum ensue on heating the core particle preparation. In conjunction with other results (1) these data suggest four states for the core particle at increasing temperatures. Additionally, analysis of the spectrum of the unmelted core particle and its differences from protein-free DNA of the same length suggests that the conformation of the phosphodiester backbone and/or its interactions with histones along the length of the core particle DNA segment may not be uniform.


Transcription of viral genes in chromatin from adenovirus 2 transformed cells by exogenous eukaryotic RNA polymerases. The transcription of chromatin from adenovrius 2 transformed rat cells by murine plasmacytoma RNA polymerases I, II and III has been studied. Both the total RNA synthesis and transcription of the integrated adenovirus 2 genes by RNA polymerase II represent de novo DNA transcription as assessed by their sensitivity to actinomycin D. It is shown that each RNA polymerase class has characteristic ionic strength activation profiles and metal ion requirements. RNA polymerase II transcribes the integrated adenovirus 2 genes in chromatin at a frequency 25- to 50-fold higher than their sequences are represented in the genome. In contrast, no detectable viral RNA is synthesized when deproteinized DNA is transcribed. In the presence of Mn2+, all three RNA polymerases (I, II and III) transcribe the integrated viral genes at approximately the same relative frequencey. However, the Mg2+ as divalent cation, the proportion of the total RNA which represents viral gene transcripts is increased 3- to 4-fold with RNA polymerase II, while it remains unchanged for RNA polymerases I or III.


Computer programs for the assembly of DNA sequences. A collection of user-interactive computer programs is described which aid in the assembly of DNA sequences. This is achieved by searching for the positions of overlapping common nucleotide sequences within the blocks of sequence obtained as primary data. Such overlapping segments are then melded into one continuous string of nucleotides. Strategies for determining the accuracy of the sequence being analyzed and reducing the error rate resulting from the manual manipulation of sequence data are discussed. Sequences mapping from 97.3 to 100% of the Ad2 virus genome were used to demonstrate the performance of these programs.


Dysfunctional uterine bleeding: diagnosis and treatment. Dysfunctional uterine bleeding is not a specific diagnosis. It encompasses a spectrum of endocrinologic dysfunction which must be considered in reference to the presence or absence of ovulation. Specific causes must not be forgotten, particularly in those women who do not respond as expected to hormonal therapy. A variety of empiric therapies are used which have no benefit, such as thyroid hormone in the absence of proven hypothyroidism, corticosteroid in the absence of proven adrenal hypofunction, and megavitamin therapies in the absence of nutritional disorders. Curettage is valuable for diagnosis, not for therapy. Medical therapy offers a successful alternative to hysterectomy in many women, but the choice must be made on an individual basis. Medical and surgical management should not be viewed as competitive treatment regimens.


The action of histamine, histamine H1-and H2-receptor agonists on noradrenaline level in rat brain. Intraventricularly administered histamine (HI) (20--1000 micrograms) decreased noradrenaline (NA) .level in rat hypothalamus. 4-Methyl-HI (4-MeHI), but not 2-pyridylethylamine (2-PEA), reproduced the action of HI. Neither HI, nor 4-MeHI induced depression of hypothalamic NA content was antagonized by HI H2-receptor antagonists, cimetidine or metiamide. Chloropyramine reversed the NA-depleting action of HI.


Giardiasis: a common cause of diarrheal disease. Giardiasis has been reported increasingly among visitors to the Soviet Union and is found at epidemic and endemic levels in the United States. The main source of infection is contaminated water. Children, homosexual males, and patients with gastrectomy, achlorhydria, hypogammaglobulinemia, secretory IgA deficiency, or alteration in immune status are particularly susceptible to severe, often chronic, infection. Symptomatic giardiasis can be acute, subacute, or chronic. Symptoms are explosive, watery, foul-smelling stools or semisolid stools with evidence of steatorrhea; flatulence; abdominal distention; and weight loss. Diagnosis usually can be established by examination of stool or duodenal fluid for cysts or trophozoites. Quinacrine hydrochloride is the drug preferred for treatment, but metronidazole and furazolidone are also useful.


Some behavioral effects of histamine H1-, and H2-receptor agonists in rats. 2-Pyridylethylamine (2-PEA) and 4-methylhistamine (4-MeHI) when given intraventricularly at doses of 30 and 100 microgram had no discernible effect on spontaneous locomotor activity and did not visibly change gross behavior of rats. 4-MeHI(100 micrograms) injected to tranylcypromine-pretreated rats increased locomotor activity and induced head twitches. Locomotor activation was not antagonized by either cimetidine or metergoline but was prevented by alpha-methyl-p-tyrosine and haloperidol and reduced by phentolamine. 4-MeHI-, and histamine (HI)-induced head twitches were not antagonized by either cimetidine or metiamide but were completely abolished by metergoline and p-chlorophenylalanine. It is concluded that 4-MeHI--induced locomotor activation and both HI-, and 4-MeHI-induced head twitches are respectively catecholamine-, and serotonin-dependent phenomena. The behavioral effects of HI and HI H1-and H2-receptor agonists are discussed with regard to possible HI interactions with both catecholamine and serotonin systems in brain.


Identification of gallstone disease.

Identification of gallstone disease. Oral cholecystography is the most widely used and accurate method for evaluating the gallbladder. Nonvisualization of the gallbladder on a two-day study is virtually diagnostic of cystic duct obstruction or gallbladder membrane dysfunction. Intravenous cholangiography may be used to confirm cystic duct obstruction but more commonly is used to demonstrate common-duct pathology. Percutaneous transhepatic cholangiography and endoscopic retrograde cholangiography are the best studies to demonstrate the intrahepatic and extrahepatic biliary tree. Tomography may be helpful in oral cholecystography and is invaluable in intravenous cholangiography. Ultrasonography appears promising for gallbladder evaluation, but reliability is greatly influenced by the equipment available and personnel employed. Biliary drainage and bile analysis are of value in patients with negative radiographic studies and clinical symptoms suggestive of biliary tract disease.


Temporal arteritis and polymyalgia rheumatica: two aspects of one disease. Temporal (cranial, giant cell) arteritis and polymyalgia rheumatica are probably different expressions of the same inflammatory disease. The disease occurs mainly in the elderly and is characterized in the early stage by vague, nonspecific symptoms. Tenderness of the temporal arteries is diagnostic but rarely is present early. Unfortunagely, loss of vision usually occurs before the diagnosis is made. A high index of suspicion and frequent testing of the ESR in elderly patients with vague complaints may lead to earlier diagnosis and appropriate therapy and thus may prevent blindness and even death.


Diesters of 4,4'-(piperazine1,4-diyl)bis-4-oxo 2-butenoic acid. The title compounds were prepared by acylation of piperazine with maleic acid ester chlorides in methylene chloride medium in the presence of K2CO3. In preliminary screening some of them showed marked antimitotic activity on plant materials.


Endogenous catecholamines during noradrenaline and adrenaline hypertension. Intravenous infusion of noradrenaline (NA) (30 micrograms/kg) results in reflectory increase of adrenaline (A), this has been established by spectrofluorometrical assay of the difference in the plasma catecholamine level during noradrenalinemia with and without elimination of circulatory reflexes. Analogically, intravenous infusion of A brings about reflectory increase of NA level. In the presence of hexamethonium a substantial amount of injected NA remains in the blood. After adrenalectomy the uptake of injected catecholamines is greatly enhanced.


Diabetes mellitus: test strategies for diagnosis and management. Fasting plasma glucose determination is the test of choice for diagnosis of diabetes. Glucose tolerance testing should be reserved for patients with borderline fasting values or possible diabetic complications and those suspected of having gestational diabetes. Strict attention to patient variables is essential if glucose tolerance testing is to be of value. The diagnosis of diabetes should be reserved for those patients with symptoms and unequivocal hyperglycemia, those with fasting plasma glucose values of 140 mg/dl or more repeatedly, and those with glucose tolerance test values of 200 mg/dl or more at two hours and at least one other time.


Reactivity of normotensive and spontaneously hypertensive rats (SHR) to some antihypertensive agents after acute and chronic treatment. Hypotensive action of some antihypertensive agents was studied on normotensive and spontaneously hypertensive rats in acute and chronic treatment. The reactivity of normotensive rats was much more weaker than in rats with spontaneous hypertension (SHR). Results of the experiments indicate also that the selection of new antihypertensive agents should be based on tests carried out with at least two models of hypertension.


Hypertensive crisis possibly due to drug interaction. This report describes a case of possible drug interaction in a patient taking chlorpromazine and guanethidine who experienced a hypertensive crisis following initiation of doxepin therapy. (We have been unable to find a report of a similar interaction at the doses described.) It is advised that physicians exercise caution when contemplating concomitant use of doxepin and guanethidine, even in usually recommended doses.


Synthesis of N-aryl-N'-[2-phenyl-3-quinazoline(3H)-4-one] acylthiourea derivatives as anticonvulsants. By the reaction of [2-phenyl-3-quinazolineo (3H)-4-one] acyl isothiocynates and appropriate aryl amines in acetone, 24 new compounds, having a substituted thiourea grouping at the 3-position of the quianozolone moiety, were prepared. All compounds, except two, showed different degree of protection against pentetrazol induced seizures test in albino mice. While studying the effect of structural variation no definite pattern could be observed due to variations in 1-aryl moiety, but it was noticed that generally the branching or lengthening of 3-acyl chain either diminishes or does not effect the activity.


Exercise in diabetes: therapeutic implications. The metabolic response to acute exercise in diabetics is largely determined by the availability of insulin. In nonketotic, insulin-dependent diabetics, acute exercise lowers the level of plasma glucose, whereas in insulin-deficient, ketotic patients, blood glucose may increase during exercise. Exercise accelerates insulin absorption from the exercising limb. Thus, by injecting insulin into a nonexercising area (eg, into the abdomen before leg exercise), the degree of exercise-induced hypoglycemia can be reduced in diabetic patients. Also, elevation in the level of antiinsulin hormones caused by exercise is exaggerated in diabetics if insulin treatment is less than optimal. This abnormal hormonal response may be restored to normal by improved methods of insulin delivery. In normal subjects, regular exercise or training results in enhanced body sensitivity to insulin. This rise in sensitivity may be mediated by augmented insulin binding to receptors. Furthermore, regular exercise may decrease levels of serum triglycerides and low-density lipoprotein cholesterol and increase those of high-density lipoprotein cholesterol. These beneficial effects of training on carbohydrate and lipid metabolism observed in normal subjects may extend to diabetic patients as well.


Severe acute mountain sickness.

Severe acute mountain sickness. The experiences of acute mountain sickness (AMS) as it has presented to a physician working in a general hospital at 1370 m in Kathmandu, nepal, are described. The features of 39 cases are analysed. It is suggested that AMS should be classified into benign and malignant forms.


Birmingham Medical Research Expeditionary Society 1977 Expedition: signs and symptoms. The BMRES group of 17 persons is described and details are given of the trek to 5400 m. The research programme is outlined. Moderately severe acute mountain sickness (AMS) was observed in 5 subjects. All subjects were rated according to their symptoms related to AMS by interview, peer review and self-assessment, and the results compared.


Birmingham Medical Research Expeditionary Society 1977 Expedition: Psychological aspects of acute mountain sickness. In order to assess the importance of personality and expectations in the development of Acute Mountain Sickness (AMS), 3 standard personality questionnaires and a Mountain Sickness Anticipation Questionnaire were completed by all of the 17 BMRES expedition members before their departure for the Himalayas. AMS could not be predicted with these tests and its occurrence, when assessed either by clinical interview or by peer review, bore no significant relationship to personality. For comparison daily self-assessment of the signs and symptoms of AMS were also conducted throughout the expedition, using graduated and graphic rating scales. The results were found to be unreliable and dependent upon personality factors. These findings have implications for those assessing others at high altitude.


Birmingham Medical Research Expeditionary Society 1977 Expedition: the diuresis and related changes during a trek to high altitude. Continuous 24-hr urine collections were made by 17 subjects during a trek to 5400 m. Fluid intake was recorded by diary. Weight and fat folds were measured daily. The results were compared with daily measurements of packed cell volume (PCV) and reticulocyte count. Early fluid retention was matched by a fall in PCV. There was a diuresis with negative fluid balance towards the end of the ascent and again early in descent. There was a slight trend for fluid retention to occur in those most affected by acute mountain sickness but the effect was not marked.


Birmingham Medical Research Expeditionary Society 1977 Expedition: effect of a Himalayan trek on whole body composition, nitrogen and potassium. The body composition of the 17 members of the BMRES was studied using body weight and fat fold calipers, with measurements of whole body potassium and nitrogen. The full assessment was made just before departure from Birmingham and immediately on return. Daily observations of body weight and fat folds were made during the trek to high altitude. During the ascent all subjects lost body fat but there was little change in lean body mass. After return it was found that there had been further loss of body fat and some loss of lean tissue also. One subject who took methandienone retained potassium but had no increase in body nitrogen. There was no correlation between changes in body composition and acute mountain sickness.


Birmingham Medical Research Expeditionary Society 1977 Expedition: serum and urine proteins during a high altitude trek. Serum and urine proteins were measured daily in 17 subjects undertaking a typical high altitude Himalayan trek. Marked changes occurred in a variety of serum proteins as a result of plasma volume alterations and 'stress'. There was only a sporadic increase in proteinuria. None of the changes was related to the development of acute mountain sickness.


Birmingham Medical Research Expeditionary Society 1977 Expedition: changes in renal function observed during a trek to high altitude. Changes in renal function were observed in 17 subjects during the course of a trek to high altitude. Comparison was made between these changes and the clinical assessment of acute mountain sickness (AMS). Periods of natriuresis occurred during ascent and descent, that during ascent being related to a fall in plasma aldosterone. Alterations in serum and urinary potassium suggested that potassium retention occurred during the ascent to altitude. No significant correlation occurred between changes in renal function and the severity of AMS before the illness being clinically apparent. When this was so, the severity of AMS correlated with a decreasing urine output, increasing positive fluid balance and a decreasing excretion of sodium and potassium; these changes were produced in part by a decrease in glomerular filtration rate.


Exercise and altitude.

Exercise and altitude. In studies concerned with the performance at high altitude it is often not possible to separate the effects which are due to hypoxia from those associated with the stress of ascent. Some of the  basic observations concerning exercise at high altitude are reviewed and a study is described which was designed to simulate the physical exertion of high altitude mountaineering but performed at altitudes below 1000 m. Some of the changes observed including peripheral oedema might under other circumstances have been ascribed to altitude.


Birmingham Medical Research Expeditionary Society 1977 Expedition: cardiopulmonary function before, during and after a twenty-one-day Himalayan trek. The heart rate response to exercise was studied in 17 normal subjects before, during and after a 21-day Himalayan trek to 5490 m. The group were fitter before the trek when compared to normal subjects of a similar age (P less than 0.001) having a lower heart rate response to exercise for their lean body mass. The trek significantly increased the 'fitness' of the group as a whole (P less than 0.025) but this was not seen in the 5 members of the group who had severe acute mountain sickness. Similar changes were noted within the first 8 days of the trek before symptoms of mountain sickness occurred. The group had normal pulmonary function before the trek but peak expiratory flow rate and vital capacity decreased with altitude. The effect was more marked in the subjects with acute mountain sickness.


[Measurement of respiratory resistance by forced oscillations in healthy children (author's transl)]. The measurement of respiratory resistance by forced oscillations, a theory known for over 20 years, has found a practical application by a simplification of the method. The comparison with the plethysmographic measurement of the resistance of airways in 27 healthy children shows a good correlation (r = 0,73), similarly with F.E.V1 (r = 0,78) and peak flow (r = 0,61). This technique does not require the forced expiration demanded by the measurement of F.E.V1 and peak flow and it does not need the costly equipment of body plethysmography. In this technique the causes of error are easy to detect. Its rapid execution and the absence of discomfort should make it a choice method in the functional respiratory tests in the child.


[Echographic study of thoracic lumps in 9 operated cases (author's transl)]. The authors report 9 cases of thoracic lumps investigated by echography in children and adults and operated later. The ulta-sonic study of intra-thoracic lumps is possible thanks to the echographic "windows" which are described; mediastinal and parenchymal lumps can thereby be investigated provided that they have a direct contact point with the chest wall. The echographic examination cannot reveal the histological nature of the lump but it is interesting in that in case of a liquid content it can assert its benignity, all with a technique simple and easy to use especially in children. The reliability of this examination seems good, the echostructure of the lesion being correctly defined in the 9 cases and in the 14 observations in the literature which are summed up.


[Clinical, pathologica and genetic analysis of 53 cases of broncho-pulmonary cancer, still surviving after 5 years (author's transl)]. The authors take up again the clinical, pathological and genetic analysis of 53 bronchial cancers with a survival exceeding 5 years. The operation samples were reexamined; a H.L.A. grouping was done in every cases. On the whole, it concerned patients under 60 in good general condition; the extension of their lesions only required a limited exeresis (lobectomy). The epidermoid form predominates, the lymph node invasion is rare. Seventy four per cent of the patients carry at least one of the following antigens: W 19, A 10, B 5, BW 35. But the study did not prove the existence of a genetic factor conditioning the survival.


[A follow up of pulmonary function after Mendelson's syndrome (author's transl)]. A follow up of pulmonary function has been performed on three patients having recovered from Mendelson's syndrome (between 1 and 70 months). In the 3 patients a syndrome of diffuse interstitial pulmonary fibrosis was observed. These perturbations disappeared relatively quickly. They were compatible with pulmonary oedema due to the disappearance of surfactant, consecutive to acid inhalation.


[The use of a laser beam in bronchology. Resection of 2 tracheo-bronchial tumours (author's transl)]. Two properties of the laser beam can be used in medicine: coagulating and sectionning. The possibility for the beam to reach its target through an optic fibre, adaptable to the broncho-fibroscope, enables its use in bronchology. Two patients with a malignant tracheo-bronchial tumour received this treatment. Results were encouraging. Some practical applications are proposed.


[Observation of a pulmonary histoplasmosis with Histoplasma capsulatum (author's transl)]. From an observation of pulmonary histoplasmosis with Histoplasma capsulatum in a Haiti woman living in France for 2 years, the authors recall the mycological and epidemiological data of this fungus as well as the main clinical radiological and biological signs of the disease. Because of its tendancy to dissemination, histoplasmosis can have a bad prognosis. It is difficult to diagnose in our countries and facing a chronic pulmonary form, a diagnosis of tuberculosis is often thought of. But its possibility is to be envisaged in case of a journey in countries of endemia, and the disease should be confirmed by: -- several samplings to trace the fungus, bringing the mycological prove of the disease; -- serum uses; -- modalities and difficulties of the treatment by Amphotericin B are also recalled.


[Mesocolic and mesenteric lymph node granulomatosis together with a sarcoidosis (author's transl)]. The authors report the case of a 22 year old woman with a recognized pulmonary sarcoidosis with disorders of the alveolo-capillary diffusion, admitted to hospital for feverish diarrhea. The discovery of right colic radiological lesions with no obvious origin, led to an exploring laparotomy which revealed large mesocolic and mesenteric adenopathies: histologically they were granulomatous lesions, of epithelioid and giant cells, tending to sclerosis. Colic and ileal mucosae were normal. From this observation the authors analyze the difficulties in diagnosing granulomatoses of the intestine and mesenteric lymph nodes and their eventual sarcoidic nature. Because of the lack of arguments in favour of tuberculosis and of the normal appearance of the intestinal coats, the sarcoidic etiology of these adenopathies appears most probable.


Incorporation of H3-glucose into the parasite cells of Eimeria tenella. The incorporation of H3-glucose into the parasite cells of Eimeria tenella was observed with electron microscopic autoradiography. H3-glucose was incorporated into the nucleus, nucleolus, mitochondria, and endoplasmic reticulum of the first schizont, but was not incorporated into the nucleus and nucleolus of young macrogametocyte.


Caloric value of cardiac response to hot environments. Single Comb White Leghorn hens were placed in a controlled temperature chamber, initially at 26 C and subsequently raised to 35 C. Recordings of the surface and rectal temperatures, respiratory rate, heart rate, and oxygen consumption were made at the 26 C basal temperature and compared with those made at the 35 C ambient temperature. Sequential doses of saline and atropine were injected following the thermoregulatory response of birds to 35 C. The thermal challenge resulted in an average increase of 53 breaths per minute in the respiratory rate during the pre-injection period at 35 C and rose steadily to the end of the thermal challenge. The saline elicited no effects on any parameter measured; however, the atropine resulted in a mean increase of 62 beats per minute in the heart rate. There was no difference in the pre-atropine heart rate at 35 C and the basal heart rate at 26 C. Following the atropine induced increase in heart rate there was a 13% increase in energy expenditure. Thermal-bradycardia in hot environments may represent a first line of defense against hyperthermia by reducing the added heat load.


Hemorrhagic syndrome of chicks produced by Penicillium citrinum AUA-532 contaminated corn. Rations containing 0%, 6.2%, and 62% Penicillium citrinum contaminated corn were fed to White Leghorn chicks for 5 weeks. The 62% ration induced hemorrhaging throughout the digestive tract which resembled the lesions observed in acute cases of the poultry hemorrhagic syndrome. The 6.2% ration caused atrophy of lymphoid tissue, anemia, leucopenia, and fatty bone marrow which resembled the lesions observed in chronic cases of the poultry hemorrhagic syndrome. The syndrome was reproduced in chicks using ethanol and chloroform extracts of the P. citrinum contaminated corn.


Digestion and absorption of lipids in chicks fed triglycerides or free fatty acids: synthesis of monoglycerides in the intestine. Digestion and absorption of lipids were determined in 3-week-old chicks fed diets containing triglycerides, free fatty acids, or free fatty acids with added glycerol. The 91YCl3 was present as a non-absorbed reference substance. Overall absorption of total fatty acids was highest in triglyceride-fed chicks and lowest in chicks receiving fatty acids. Net secretion of endogenous fatty acids from biliary phospholipids was observed in the duodenum of triglyceride fed chicks, whereas in the other treatments net absorption was observed at this site. Fractionation of the intestinal lipids by thin layer chromatography showed that triglycerides were almost completely hydrolyzed in the duodenum. Monoglycerides were present at high levels in the duodenum of triglyceride fed chicks and at lower levels in chicks receiving free fatty acids and glycerol and at trace levels in free fatty acid-fed chicks. Total micellar fatty acids were lowest in the duodenum of free fatty acid-fed chicks. In vitro incubation of pancreatic homogenate with oleic acid, glycerol, phosphatidyl choline, and taurocholate revealed synthesis of monoglycerides. The poorer fat absorption observed on feeding acidulated soapstock instead of triglycerides is partially explained by less efficient micellarization when free fatty acids are fed.


The response of three strains of fowl to alpha-chloralose. A heavy and two light strains of pullets suffered a decline in egg production but an improvement in body weight gains when alpha-chloralose, a tranquilizer, was included in their diets at .1 to .3%. Feed efficiency also experienced a decline while egg size and Haugh unit were not affected.


Hematology of Single Comb White Leghorn pullets fed aflatoxin-contaminated and ammonia-treated corn. Erythrocyte count, hemoglobin content, percent hematocrit, or mean corpuscular hemoglobin were not affected by feeding aflatoxin levels of 2.3 ppb and 500 ppb to Single Comb White Leghorn hens. Pullets fed ammoniated corn without aflatoxin and pullets fed corn containing 500 ppb aflatoxin in the finished feed had significantly reduced mean corpuscular volume.


Effects of diazacholesterol, triparanol, and beta-sitosterol on egg cholesterol deposition in coturnix quail. Studies were undertaken to determine the effect of inhibitors of cholesterol synthesis on deposition of cholesterol in eggs of Japanese quail. Results indicate that this bird responds similarly to the laying hen, making it a useful screening device for these types of compounds. Administration of either triparanol or 20,25 diazacholesterol resulted in a decreased cholesterol content of the yolk. Concomitant with this decrease was an increase in desmosterol deposition. Beta sitosterol (2%) fed either alone or with lecithin (2%) did not result in a decrease in egg yolk cholesterol. No beta-sitosterol was found in the egg yolk. Diazacholesterol fed either with sitosterol, or sitosterol plus lecithin, was not effective in reducing the total sterol content of egg.


Effects of furazolidone in the early chick embryo. Effects of furazolidone in early chick embryos were investigated in order to determine the feasibility of using this biological system as a model for studying the mechanism(s) of furazolidone toxicity. Double and triple injections of varying dosages of furazolidone were administered via the yolk sac at early stages of incubation. Toxic levels of the drug affecting both growth and levels of glycogen were dosage-dependent and time-dependent. Double injections of furazolidone at a dose as little as 60 microgram/egg administered at 40 and 72 hr of incubation significantly retarded growth (P less than .001) and increased glycogen levels (P less than .05) in the embryo and yolk sac. Mortality was high in embryos receiving toxic levels of furazolidone, but no gross abnormalities were observed.


[Operability and fitness for anesthesia (author's transl)]. The decision as to operability and fitness for anaesthesia must be made jointly by the surgeon and the anaesthesiologist and only by them. They arrange for the required diagnostic and therapeutic measures to be taken to assure the success of the operation. The advice of other specialists as to the treatment of disorders unconnected with the primary disease does not extend to questions regarding operability and fitness for anaesthesia. The preparation of out-patients for surgery requires particular care on the part of the anaesthesiologist. A detailed pre-operative explanation to the patient of what is involved will greatly help to relieve the patient's anxiety.


[Indications for surgery and operability from the point of view of the surgeon (author's transl)]. The decision as to surgical intervention and operability must also take into account somatic, personal, psychosomatic and social factors. The advances made in medical science have added to rather than detracted from the difficulties of the problem. Although the final decision rests with the surgeon the indications as to surgical intervention and operability present problems which demand interdisciplinary co-operation, first with the general practitioner, then, after admission to hospital, with the various specialists and subsequently again with the general practitioner.


[An attempt to determine optimum dosage of fentanyl in neurolept analgesia (author's transl)]. In a retrospective study a comparison was made of the doses of Fentanyl used by anaesthetists to induce and maintain neurolept analgesia for a variety of surgical operations (158 cases). Dosages differed widely both for different surgical procedures and for different anaesthetists. A method, based on pharmacokinetic considerations, was developed for calculating Fentanyl requirements during any stage of anaesthesia. The dose/time relation, as represented by y = At + B (1--e-kt) makes it possible to calculate the required doses of Fentanyl; this enables the anaesthetist to maintain a stable level of anaesthesia and makes antagonization of Fentanyl unnecessary. Methods for determining the coefficients of the dose-time equation are described. Simulation by an analogue computer showed that by using the suggested procedure substantial variations of Fentanyl concentration in the brain and other body compartments can be avoided.


[Tetanus-Myomechanographic measurement of receptor liberation by pyridostigmine in neuromuscular block produced by d-tubocurarine, gallamine, pancuronium or alcuronium (author's transl)]. In 48 patients anaesthetized with nitrous oxide-oxygen and hexobarbitone the neuromuscular (n-m) block (greater than or equal to 95% depression) produced by roughly equipotent doses of d-tubocurarine (dTC), gallamine (GALL), pancuronium (PANC) or alcuronium (ALC), respectively, was antagonized by 10 mg of pyridostigmine (P) applied intravenously 35-460 min after the relaxant at variable levels of spontaneous recovery from n-m block. Muscular reactions to tetanic stimulation (30 to 400 Hz, 4-5 s each) of the ulnar nerve transmitted by a force-displacement system served as a measure for calculating the relative amount of n-m receptors liberated from relaxant molecules. Within 3-10 min after its injection P increased the number of relaxant-free n-m receptors by 16 +/- 6% (M +/- SD). Thereafter recovery progressed at similar speed as before. Reinjections of 5-10 mg P were comparably as effective as the first injection. No correlations were to be found between the effectiveness of P and the dose of relaxant applied (r = 0,12 to 0,27), the level of recovery reached before P (r = 0,32), or the time at which P was injected after the relaxant (r = -0,39), respectively. However, the amount of receptors liberated by P decreased with increasing recovery from n-m block and with increasing time interval between the relaxant and the antidote injection. P was significantly more effective (P less than 0,01), when applied within 150 min after the relaxant than at applications after that time. The relative number of receptors liberated by this drug was insignificantly larger in the PANC-and GALL-block than in the ALC-and dTC-block.


[Enflurane anaesthesia in a "clinically closed" system (author's transl)]. The following report will deal with the application of "Enflurance" in the clinically closed (rebreathing) system. The term clinically closed system defines a procedure where the phase of approach up to MAC and the phase of subsidence is pursued in a half-closed system and the maintenance in a closed system. Enflurane was chosen as volatile anaesthetic, because it has been increasingly applied world-wide on account of its convenient qualities. Besides enormous savings by way of a low consumption of narcotics, this method offers a reduced environmental pollution. In case of sufficient monitoring, this technique proves attractive to smaller hospitals, disaster medicine and central hospitals likewise.


[Effects of general anaesthesia with halothane and neuroleptic anaesthesia on free plasma amino acids (author's transl)]. Changes of plasma amino acids in animal studies were observed after administration of different narcotic drugs. In a randomised study the effects of neuroleptic anaesthesia and general anaesthesia with halothane on free plasma amino acids in respect to liver toxicity were investigated in patients during microsurgery of the middle ear. 24 hours postoperatively in both groups a significant increase of branched chain amino acids and of phenylalanine as metabolic sign of stress was found. Typical changes of amino acids as shown in hepatic failure could not be seen in both groups. The study showed that none of the narcotics used had a toxic effect on liver function as measured by the levels of free plasma amino acids and that for disagrees with the results of animal studies.


[A new electronic spirometer with automatic drift compensation and its clinical performance (author's transl)]. An electronic spirometer is described which, by means of a device for automatic drift compensation, allows the pulmonary function of artificially ventilated persons to be analysed continuously. The results obtained with the apparatus in artifically ventilated premature births, newborns and infants are discussed.


[Measurement of resistance and deadspace of endotracheal tubes used in paediatric intensive care medicine (author's transl)]. It is reported about the evaluation of resistances and deadspaces of endotracheal tubes. The results show that the resistances of the tubes are similar to the resistances in the patient lungs. The deadspaces are small as compared to the physiological values. It is suggested that a patient with a endotracheal tube has a smaller deadspace than one without a tube.


[Transcutaneous measurement of PaO2 by the "TCM 1" electrode (author's transl)]. Arterial oxygen pressures were compared with oxygen pressures measured transcutaneously with the aid of "TCM 1", an apparatus manufactured by Radiometer. At normal oxygen partial pressures the transcutaneously determined values for pO2 ran parallel with those measured intraarterially, but were on average 10.7% lower than the latter. In cases where oxygen partial pressures were above or below normal the figures did not agree. In these conditions, therefore, determinations in the arterial blood cannot be dispensed with. Halothane in concentrations of 0.3-0.5% and nitrous oxide (66%) in the inspired air did not affect the transcutaneous determination of pO2, but the method failed during surgical electrocoagulation.


[Antipollution device for the Jackson-Rees-system (author's transl)]. Antipollution-devices in T-piece-systems are as far useless as they convert the system. A new device for the Jackson-Rees system is described, which does not disturb its function and which may be used in fresh-air-climatisated rooms.


[Siphonaptera/fleas (author's transl)].

[Siphonaptera/fleas (author's transl)]. Fleas are small, reddish-brown, wingless insects with a laterally compressed body and a pronounced third pair of legs adapted to leaping. Of the 100 species found in Middle Europe, hardly a dozen are of medical importance, they concern mainly people in contact with domestic animals. The cat flea, Ctenocephalides felis felis, and the bird flea. Ceratophyllus gallinae, are the most important human-pathogenic species in our region. A flea bite shows first as a haemorrhagic punctum, accompanied by itching, and leads to an erythema with or without central wheal. After 12--24 hours a papule appears which persists up to 2 weeks. Linimentum zinci with 10% Neocid alleviates the itching and prevents further infestation. The fleas are destroyed in their hiding places and on their animal host by applying Toxical-, Neocid- or Noflo-powder. The tropical sand flea, Tunga penetrans, is a permanent ectoparasite of man. It is seen in people returning from the tropics. Fleas may, even in our region, be vectors of bacteria, viruses, rickettsiae and intestinal parasites.


[Pinworm infection in kindergartens of Basel (author's transl)]. Within the scope of mass examinations 949 children from kindergartens in Basle were submitted to the scotch tape test of Oxyures. The findings were positive in 64 cases (6,7%). The parents of 50 children accepted the suggestion that their children be treated twice with 100 mg of ciclobendazole each, at a weekly interval. This therapy resulted in complete cure of the oxyuriasis in all treated cases. Despite the impressive and safe action against this disease of ciclobendazole it is recommended in any case to take a second tablet, since the risk of reinfection is high.


[Dynamics of various indicators of natural immunity in diabetes mellitus]. Patients with diabetes mellitus displayed a suppression of humoral and cellular mechanisms of nonspecific factors of immunity protection: of bacteridical serum activity, of phagocytic neutrophil activity, and a sharp disturbance of profound skin autoflora. The amount of serum lysozyme diminished only in young patients with severe form of diabetes mellitus. In other patients the muramidase content was increased. Compensation of the disease led to an even greater elevation of this enzyme, and produced no significant influence on other natural resistance parameters.


[Adrenocorticotropic and somatotropic functions of the pituitary gland in juvenile thyrotoxicosis]. The adrenocorticotropic (ACT) activity of the hypophysis (according to the blood 11-HCS level) was studied in 27 female patients suffering from thyrotoxicosis, aged from 16 to 24 years (19 with moderately severe and 8 with severe form) and in 7 healthy girls. The ST activity of the hypophysis was ascertained in 11 patients by the blood STH content before and after the insulin hypoglycemia. The metopyron test was also carried out in 6 patients. The ACT function of the hypophysis proved to be increased, and the ST function was normal in the patients with thyrotoxicosis at rest and on fasting stomach. The value of the corticotropic and somatotropic response to hypoglycemia was similar in sick and healthy persons. In the patients with a pronounced elevation of the 11-HCS and STH content under the effect of hypoglycemia the metopyron response was determined by the initial adrenal cortex function.


[Clinico-morphological comparisons of autoimmune processes in the thyroid gland in the endemic goiter region]. A total of 2558 patients (95% of patients who sustained strumectomy in the Chelyabinsk region in 1969--1977 for euthyroid nodular goiter) were examined. There proved to be a relationship between the iodine content in the food products of the population and the frequency of the morphologically-definable autoimmune processes in the thyroid gland; this dependence was direct for the marked autoaggression processes, and indirect--for the initial ones. This dependence is also confirmed by a rare combination of multinodular goiter with diffuse autoimmune thyroiditis and a frequent one--with the initial autoaggression forms.


[Study of the possibility of using the "artificial pancreas" in the treatment of diabetes mellitus]. A short survey of the literature data on the use of an artificial pancreas in the treatment of patients suffering from diabetes mellitus is presented. These data point to the high efficacy of the artificial pancreas in diabetic patients, this confirming the author's personal observations.


[Effect of hydrocortisone on the state of liver chromatin in rats of different age]. Hydrocortisone (5 mg/100 g) injected intraperitoneally induced redistribution of hepatic chromatin fractions in pubertal (aged 6--8 months) and old (aged 24--26 months) rats. Changes in the content of these chromatin fractions begin earlier and end later in old animals. Hydrocortisone stimulated 14C-orotic acid incorporation into RNA of active and inactive hepatic chromatin in rats of both age groups. The maximal incorporation of 14C-orotic acid into RNA of inactive chromatin in pubertal rats occurred 7 hours after the hormone administration, and in old rats after 4 hours; in case of active chromatin these figures were 17 and 6 hours, respectively.


[Blood serum immunoglobulins in small boys and adolescents with hypogonadism]. The immunoglobulin (Ig) concentration was studied in the blood serum of 31 boys with secondary hypogonadism. There was revealed a marked fall in the content of Ig of the A, M, and G classes, particularly of the M class, in all the patients before the treatment. After disappearance of the clinical hypogonadism signs and normalization of the blood testosterone level under the effect of chorionic gonadotropin therapy the plasma Ig concentration was found to be restored to the normal; the greatest relative elevation was revealed in case of IgM. Possible relations of testosterone with the immunocompetent organs and tissues are discussed.


[Diagnostic and prognostic value of the glucose tolerance test in doubtful cases]. Many-year observation over the changes in glucose tolerance test of different type in 964 women, aged from 17 to 57 years, on free diet and free regimen, was carried out. The changes in glucose tolerance were the more unfavourable--the greater were its deviations during the initial examination. The doubtful tolerance changes, diagnosed by hyperglycemia on fasting stomach, with the normal blood glucose level one and two hours after glucose load, were identical to the normal tolerance changes, serving as a variant of the latter.


[Effect of chronic prostatitis on the endocrine activity of the testes]. It was shown in experiments on rabbits that androgenopoiesis was disturbed in the testes under conditions of aseptic chronic inflammation of the prostate: against the background of a sharp reduction of testosterone and androstendion formation there was observed production and discharge into the blood of great amounts of progesterone, this being not characteristic of the incretory activity of the intact gonads. It is suggested that the search for the ways of steroidogenesis restoration in the testes under conditions of induced pathology should be directed to the activation of enzymes catalyzing the processes of progesterone transformation into 17-alpha-hydroxyprogesteron, androstendion, and testosterone.


[Change in the level of acid-soluble adenine nucleotides in rat tissues in experimental hypo- and hypercorticism]. The level of free adenine nucleotides was studied in the rat tissues in deficiency and excess of the adrenal cortex hormones in the organism. Hypocorticism induced by adrenalectomy led to reduction of the concentration of total and individual adenine nucleotides in all the tissues examined, except ADP in the spleen and AMP in the myocardium. Single administration of hydrocortisone to intact rats caused an increase in the level of total and individual adenine nucleotides in the liver and reduction of their concentration in the spleen and the skeletal muscle. Along with a fall of total nucleotides and ADP concentration in the myocardium, the AMP content increased and the ATP remained unchanged. Experiments carried out pointed to the participation of glucocorticoids in the metabolism of free tissue adenine nucleotides. Specificity of the reaction of individual tissues to the changes in glucocorticoid hormones level in the organism was noted.


[Role of glucocorticoids in the mitochondrial DNA replication]. The liver mitochondrial DNA biosynthesis was found to be enhanced in adrenalectomized rats. Cortisol administration to adrenalectomized rats caused a decrease of the liver mitochondrial DNA biosynthesis. Sedimentation analysis of the mitochondrial DNA in CsCl--EtBr gradient has demonstrated that 3H-thymidine incorporation into the mictochondrial DNA increased mainly in the "open circular" molecules fraction.


[Insulin receptors in isolated cells of murine lactating mammary gland]. Insulin receptors in isolated cells from lactating mouse mammary gland were investigated by 125I-labeled insulin. A modified chloramin T-method after Hunter and Greenwood (1962) was used for insulin labelling with 125I; 125I-insulin with a high specific activity (80--120 muCi per microgram), high immunoreactivity and preserved biological properties was obtained. Three methods of labeled insulin purification were also studied: absorption on the cellulose column, gel chromatography on different types of Sephadex (G-25, G-75, G-100) and electrophoresis on polyacrylamide gel. The best conditons for iodination and purification of 125I-insulin suitable for studying the receptor-insulin binding in the isolated cells from mouse mammary gland were selected. Insulin binding to its receptor was found to be a specific reversible process, having high affinity and a tendency to saturation. The receptor-insulin-binding equilibrium was reached in 30 min at 24 degrees C. Specific binding was observed at a very low concentration of 125I-insulin (0.4.10(-9)M), close to the physiological level. Saturation was observed at a concentration of over 1.5.10(-9)M. Native, non-labeled insulin, at a concentration of 1 ng per ml lowered the labeled insulin binding by 10--20 per cent, whereas 10 ng per ml led to a 60 per cent lowering. The affinity constant of the process was about 10(9).M-1. Each cell had about 3 000 to 4 000 insulin binding sites.


The use of Doppler shift radar to monitor physiological and drug induced activity patterns in the rat. Doppler shift radar was used to monitor circadian activity patterns in the rat and to study the behavioural effects of p-chloroamphetamine and d-amphetamine. Activity was classified in two ways:-(a) slow (non-locomotory) and high (locomotory) speed movements. (b) the number of starts of activity, within either the slow or high speed zones of activity during a pre-set time. p-Choloramphetamine (5 mg/kg) produced a biphasic activity response; an initial increase in continuous non-locomotory activity followed by a longer lasting increase in exploratory locomotion containing regular starts of activity. d-Amphetamine (2.5 mg/kg) produced an increase in both non-locomotory and locomotory movements but a marked reduction in starts of activity (i.e. continuous non-exploratory activity). The combination of information on the amount and pattern (starts) of activity allows a more detailed analysis of the effects of drugs on activity to be made than with existing automated methods.


LiCl-induced selective depression of saccharin drinking in the mouse. Water-deprived mice were injected with various concentrations of LiCl or NaCl 15 min before they were allowed to drink either water or 0.1% saccharin. The NaCl injections produced a dose-dependent increase in intake of both fluids: however, the higher dosages of LiCl produced a selective depression of fluid intakes. Saccharin intakes were depressed for less than one hr but water intakes were not affected. LiCl injections also depressed general activity and produced an apparent shift of water from blood into cells. The LiCl-induced depression of saccharin was not significantly influenced by extensive previous experience with the drinking fluid.


On the action of nicotine and cotinine on central 5-hydroxytryptamine neurons. The actions of nicotine, and its main metabolite cotinine, on 5-hydroxytryptamine (5-HT) neurons in the brain of the rat have been assessed biochemically (on turnover, uptake, release, overflow and binding of 5-HT in brain) and functionally (on extensor reflex activity, which is 5-HT dependent). Nicotine and cotinine in repeated doses of 2 mg/kg caused a reduction of brain 5-HT turnover, which was not blocked by pretreatment with mecamylamine, and nicotine sifnificantly inhibited the effects of norfenfluramine and 5-methoxydimethyltryptamine on extensor reflex activity, effects counteracted by mecamylamine. In low concentrations cotinine weakly inhibits the uptake and retention of 5-HT and also increases its spontaneous release in vitro. The biochemical findings suggest that the reduction of 5-HT and also increases its spontaneous release in vitro. The biochemical findings suggest that the reduction of 5-HT turnover caused by high doses of nicotine are mediated, at least in part, by its main metabolite cotinine. The experiments on extensor reflexes indicate that nicotine can block the functional expression of 5-HT receptor activity in the spinal cord by an action beyond the 5-HT receptor at nicotine-like cholinergic receptors whose location is also discussed.


Lateralisation of function in the chicken fore-brain. There is lateralisation of function in the chicken fore-brain. This was revealed by examining the behavioural modifications produced by administration of cycloheximide into the left or right hemisphere on Day 2 of post-hatched life. Visual discrimination learning of a task requiring a search for food was found to be performed either entirely or, at least, to a greater extent by the left hemisphere. Visual habituation learning was not found to be lateralised. The left hemisphere is more involved in auditory habituation than is the right; administration of cycloheximide to the left hemisphere slowed auditory habituation, as did bilateral administration, but treatment of the right hemisphere was ineffective. There are indications that the right hemisphere plays a more important role in response to novelty. A side-preference for response to stimuli seen by the left eye was demonstrated. These results are discussed with reference to head orientation during development in the egg.


Ascending 5-HT pathways and behavioural habituation. Microinjections of 5,7 -dihydroxytryptamine into both the dorsal and median raphe nuclei resulted in 70-85% depletions in striatal and hippocampal 5-HT concentrations but did not affect habituation of orienting in the lick-distraction test, habituation of activity in an open-field or habituation of exploration in the holeboard test. Lesioned animals were hypoactive in the latter two tests and defaecated more than control rats in the open-field suggesting an increase in emotionality or fear. Rats with selective 5,7 -dihydroxtryptamine lesions of either the dorsal or the median raphe nucleus also showed no impairment of habituation of orienting or exploration. However, median raphe lesioned animals were hyperactive at some stages of the holeboard test. In contrast to previous studies, the results suggest intact ascending 5-HT pathways are not necessary for behavioural habituation of orienting, activity or exploration. Rather, 5-HT neurones may be involved in modulation of activity or responsiveness to aversive environments.


Juvenile-onset obesity and deficits in caloric regulation in MSG-treated rats. Caloric regulation and the development of obesity were examined in rats which had received parenteral injections of monosodium glutamate (MSG) as neonates. Rats were injected with either 2 mg/g or 4 mg/g MSG on alternate days for the first 20 days of life. Lee Indices of obesity were calculated at 22, 70, and 130 days of age. Animals in the 4 mg/g group were significantly more obese than controls at all three ages. However, both food intake and body weight of this group were significantly lower than those of controls. In adulthood, the ability to regulate caloric intake was tested by allowing animals access to diets of varying caloric densities. While control animals maintained relatively constant caloric intakes across dietary conditions, MSG-treated animals demonstrated an inability to respond to caloric challenges. Treated animals decreased caloric intake on a diluted diet and consumed more calories than controls when presented with a calorically dense diet. This inability to regulate caloric intake is compared with regulatory deficits observed in animals sustaining lesions of the ventromedial hypothalamus. The value of an animal model of juvenile-onset obesity is also discussed.


Development of motor activity in young rats following perinatal methadone exposure. Ambulatory behaviors of 21, 45, and 60 day old rats exposed to methadone (5 mg/kg) during gestation and/or lactation were studied by assessing locomotion in an activity cage, open field, and activity wheel, and latency times to step down from an elevated platform. Methadone-exposed rats were found to be generally less active than controls at 21 days of age and more active than saline-treated pups at the 45 and 60 day test periods. In addition, behavioral responses appeared to be dependent on the timing and duration of opioid treatment. These data suggest that prenatal and/or postnatal methadone exposure affects behavior in young rats and provide a functional correlate to our earlier observations of microscopic and neurochemical changes in the brains of methadone-treated offspring.


Effects of a fixed time schedule and body weight on ethanol self-administration. Two experiments are reported. The first study showed that the patterns of ethanol acquisition using the schedule induced self-injection paradigm, are more similar to those of heroin than methadone or nicotine but dissimilar to that of d-amphetamine. The second study showed that ethanol intake once established was not maintained by removal of the schedule. The results also showed that schedule induced behavior may be stressful as indexed by a significant increase of plasma corticosterone levels.


Facilitation of conditioned motor suppression by microinjections of dopamine in the caudate nucleus of cats. The effects of Caudate Nucleus (CN) injections of catecholamines on the suppression of a motor conditioned response (MCR), lever pressing, was investigated. Cats were trained to press a lever to obtain 0.5 ml of milk when a conditioned discriminative stimulus was on (CS-on-reward on, MCR) and to suppress the response when the light was off (CS-off-reward off, suppression of motor conditioned response, SMCR). The bilateral application of 5 or 10 micrograms of Dopamine (DA) through chronically implanted cannulae in the CN significantly decreased the lever pressing in the non rewarding situation without changing the MCR. Injection of 5 micrograms of L-DOPA caused very small effects assessed during the following 10 min. However, 10 micrograms of L-DOPA produced a significant decrement of lever pressing (CS-off) in two out of four injections. These findings further support the postulation that catecholamines in the CN have a behavioral inhibitory action upon a motor conditioned response.


Differential effects on learning by ventromedial vs lateral hypothalamic posttrial injection of substance P. The effects of post-trial injection of substance P (SP) into the lateral hypothalamus (LH) and the ventromedial hypothalamus (VMH) on passive avoidance learning was studied in rats. In the VMH, 50 ng and 500 ng SP influenced neither learning of a step-down avoidance nor of an alcove avoidance response. In contrast to these findings, 500 ng SP injected into the LH significantly enhanced retention of the alcove avoidance task. Similarly, in the step-down avoidance experiment, learning was strongly facilitated by posttrial injection of 50 ng as well as 500 ng SP into the LH. These results, together with our previous data showing amnesia with posttrial injection of SP into amygdala and substantia nigra, suggest that exogenously applied SP influences the activity of those brain regions shown to contain high densities of SP-positive nerve terminals. Interestingly, the effects of posttrial SP injection parallel the effects of post-trial electrical brain stimulation on passive avoidance learning. Hence, posttrial SP retroactively facilitates or impairs learning depending on where in the brain it is injected.


Toxic interactions of ethanol with other central depressants: antagonism by naloxone to narcosis and lethality. The effects of naloxone on narcosis and/or lethality induced by diazepam, lithium, methaqualone and phenobarbital either alone or in combination with ethanol were studied in mice. Interaction toxicities between ethanol and the various psychotropic drugs were dose-dependent and so was the degree of antagonism by naloxone. Treatment with phenobarbital (10 mg/kg) or methaqualone (50 mg/kg) or lithium (4 meq/kg) prolonged the narcosis induced by ethanol (5 g/kg) by 45, 269 and 107% respectively. Naloxone (10 mg/kg) shortened the ethanol (5 g/kg) induced narcosis by 38%. Naloxone (10 mg/kg) also shortened narcosis induced by ethanol (5 g/kg) in combination with phenobarbital (10 mg/kg) or methaqualone (50 mg/kg) or lithium (2meq/kg) by 31, 12 and 38% respectively. At 10 mg/kg of naloxone, the LD50 due to methaqualone was increased from 240 mg/kg to 416 mg/kg, and the LD50 due to ethanol was increased from 9.2 g/kg to 10.8 g/kg. Multiple injections of naloxone significantly (p less than 0.01) protected against the lethality of phenobarbital but not that of lithium. These findings provide further evidence of naloxone antagonism towards various CNS depressants.