Analysis of neoplasms induced by Cas-Br-M MuLV tumor extracts

Cas-Br-M is an ecotropic murine leukemia virus isolated from wild mice that induces a wide spectrum of hematopoietic neoplasms, including T and B cell lymphomas, myelogenous leukemias, and erythroleukemias. The purpose of this study was to determine if the induction of neoplasms belonging to multiple lineages was due to the ecotropic virus itself or to the generation of cell lineage-specific recombinant viruses. The results demonstrate that in some instances (two of 12 tumor extracts tested), recombinant viruses can be recovered from primary Cas-Br-M-induced tumors that will induce lymphomas of single lineages in mice inoculated as newborns. One of these viruses is a recombinant mink cell focus-forming virus that induces T cell lymphomas, and the other is a replication-defective, fibroblast-transforming virus that induces early B lineage lymphomas in mice. Histologic and flow microfluorometric cell surface antigen analyses of primary and in vitro adapted tumors are presented in support of a modified scheme of hematopoietic cell development.     

Spontaneous deletion formation at the aprt locus of hamster cells: the presence of short sequence homologies and dyad symmetries at deletion termini.

To examine the factors governing the generation of DNA sequence rearrangements in mammalian somatic cells, we have cloned and sequenced novel junctions produced by six spontaneous deletion mutations at the aprt locus of Chinese hamster ovary cells. Our analyses indicate that these rearrangements were produced by non-homologous recombinational events occurring between short (2-7 bp) sequence repeats at the two termini of the deletion which leave one copy of the repeat in the mutant gene. Certain tri- and tetranucleotides recur at the deletion termini, suggesting that these may possibly be a recognition sequence for an enzyme involved in the event. No other gene structural alterations were found at the novel junctions or in neighbouring sequences. The deletions are not randomly distributed over the aprt gene; four termini clustered in a 40-bp sequence. This region of aprt is unusual as it contains both significant stretches of dyad symmetry which could potentially form stable DNA secondary structures and short direct repeats. Regions of dyad symmetry were also found at at least one terminus of all the deletions. In view of the similar properties of this set of deletions, possible mechanisms for the formation of this type of gene rearrangement are considered.     

Evidence for the amino acid sequence of porcine pancreatic elastase

The preparation and purification of tryptic peptides from aminoethylated Dip-elastase and [(14)C]carboxymethylated Dip-elastase, and of peptic peptides from native elastase is described. A summary of the results of chemical studies used to elucidate the amino acid sequence of these peptides is presented. Full details are given in a supplementary paper that has been deposited as Supplementary Publication SUP 50016 at the National Lending Library for Science and Technology, Boston Spa, Yorks. LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1973), 131, 1-20. These results, together with those from previously published papers, are used to establish the complete amino acid sequence of elastase, which is a single polypeptide chain of 240 residues, molecular weight 25900, containing four disulphide bridges.     

Comparison of cell-walls of Lolium multiflorum with cotton cellulose in relation to their digestion with enzymes associated with cellulolysis

Activities of several enzymes associated with cellulolysis were compared using as substrates cell-walls of Lolium multiflorum and cotton cellulose. Purified enzymes C₁ (see Ref. 1 for definition), C._x (CM-cellulase) and β-glucosidase were employed as well as culture filtrates containing C_x. Activities were determined by ability to digest the substrates and to release H₂O-soluble phenolic compounds from the grass cell-walls. The culture filtrates most active on cotton cellulose were obtained using the fungi Trichoderma viride and Fusarium solani; with grass cell-walls the most active were from T. viride, Gliocladium roseum, a species of Basidiomycetes, and one strain of Myrothecium verrucaria (IMI Strain 25 291). For the crude enzyme preparations tested, there were highly significant correlations between the digestibility of grass cell-walls and the UV-absorption of the filtrate at λ_max 290 nm and at λ_max 324 nm but there was no significant correlation between the digestibility of grass cell-walls and that of cotton cellulose. Partially purified C₁ and C_x from two different fungal sources showed activity on both substrates. Differences in MW of the H₂O-soluble phenolic compounds obtained by treatment of grass cell-walls with C₁ and C_x components suggest that these enzymes could have different modes of action. Synergism between C₁ and C_x from T. koningii occurred with both substrates but with C₁ and C_x from F. solani synergism only occurred with cotton cellulose.     

The primary structure of porcine pancreatic elastase: The N-terminus and disulphide bridges

1. The amino acid composition and N-terminal groups of purified elastase show that it is a single peptide chain of 234 residues. 2. The N-terminal sequence is Val-Val-Gly-Gly-Thr-Glu-. 3. The sequences around the four disulphide bridges were determined by using a ;diagonal' electrophoretic technique. 4. These four bridges are homologous with the four common to bovine trypsin and chymotrypsin. 5. Out of 83 residues of the elastase sequence so far determined, 43 are homologous with similar regions of trypsin and chymotrypsin. 6. The evolutionary ancestry of these enzymes is discussed.     

Respiratory depressant effects of GABA alpha- and beta-receptor agonists in the cat

The aim of this study was to evaluate the cardiorespiratory effects of intravenously administered gamma-aminobutyric acid (GABA) alpha-(4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol, THIP) and beta-(baclofen) receptor agonists and to locate the site of action of these drugs in the brain. THIP and baclofen were administered to alpha-chloralose-anesthetized cats while minute ventilation (VE), arterial blood pressure (AP), and heart rate were monitored. THIP, in doses of 0.5 to 2 mg/kg decreased VE, tidal volume (VT), and AP. No changes in respiratory rate (f) or inspiratory (TI) or expiratory (TE) duration were observed. Baclofen, in doses of 0.5 to 4 mg/kg, decreased VE, f, and AP. VT and TI increased and an "apneustic" breathing pattern was seen. THIP (9.5 micrograms), applied bilaterally to the glycine-sensitive area of the ventral medulla, reproduced the effects seen with intravenous administration. Application of 10 micrograms of bicuculline bilaterally to this area reversed the effects of intravenous THIP but not those of baclofen. Baclofen (5.6-56 micrograms), administered by the intracisternal route, produced the same respiratory effects seen with intravenous administration. We conclude that activation of GABA alpha- and beta-receptors produces cardiorespiratory depression. However, this is accomplished by different mechanisms and by actions exerted at different central nervous system sites.     

slit: an EGF-homologous locus of D. melanogaster involved in the development of the embryonic central nervous system

A family of loci homologous to the EGF-like portion of Notch, a gene involved in neurogenesis, have been identified in D. melanogaster. The sequence, spatial, and temporal distribution of both RNA and protein of one of these loci suggest a possible role in the development of the central nervous system (CNS). In situ hybridization and antibody staining of embryos show initial localization in cells along the midline of the neuroepithelium. High level expression is restricted in the developing embryo to a subset of six midline glial cells abutting growing axons. Extracellular localization is suggested by the presence of EGF-like repeats in the deduced protein sequence and antibody staining. Cytological, immunocytochemical, genetic, and molecular data show that this gene corresponds to the slit locus. Mutations in this locus result in the collapse of the regular scaffold of commissural and longitudinal axon tracts in the embryonic central nervous system.     

Antifeedant tests with larvae of Pseudaletia unipuncta: variability of behavioral response

The egg development of the African bushcricket Ruspolia differens is shown to be direct, without any delay, and to be highly synchronised. It is also strongly temperature dependent and closely fits the thermal summation equation Days=(T°C–14.8)/252, even if incubation temperatures (T) are changed during the course of development. In contrast egg development in the closely related Southern European Ruspolia nitidula is subject to a form of diapause after anatrepsis but its other developmental characteristics are very similar to R. differens. Diapause in R. nitidula may disappear in time without change of temperature and is thus unlike any diapause reported for other Tettigoniidae. Chilling diapausing eggs progressively synchronises and advances hatching, the full effect being achieved in 8 weeks at 8°C. Development in post-diapause eggs is independent of diapause treatment.The effect of partial cooling is clearly shown by the bimodality of hatching curves and a method of analysing and comparing these is discussed.The diapause syndrome acquired by R. nitidula is seen as a method by which this one species of a primarily tropical genus has been able to move into temperate zones. The role of this type of diapause in the phenology of R. nitidula is discussed.

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Zusammenfassung Die Eientwicklung der afrikanischen Laubheuschrecke Ruspolia differens (Orthoptera, Tettigoniidae) verläuft direkt, ohne Verzögerung, und ist sehr gut sychronisiert. Sie ist streng abhängig von der Temperatur und entspricht gut der Wärmesummenregel Tage=(T°C–14,8)/252 und zwar sogar dann, wenn die Zuchttemperatur (T) während der Entwicklung verändert wird. Im Gegensatz dazu durchläuft die Eientwicklung der nah verwandten südeuropäischen Ruspolia nitidula eine Art Diapause nach der Anatrepsis. Die übrigen Entwicklungsschritte sind sehr ähnlich mit R. differens. Die Diapause kann bei R. nitidula rechtzeitig ohne Temperaturwechsel verschwinden. Damit unterscheidet sie sich von den übrigen bei Tettigoniiden beschriebenen Diapauseformem. Durch Abkühlen wird das Schlüpfen diapausierender Eier synchronisiert und beschleunigt und zwar wird der volle Effekt nach 8 Wochen bei 8^o erreicht. Die Etwicklung der Nachdiapauseeier ist unabhängig von der Diapauseent-wicklung.Der Effekt partieller Kühlung wird klar ersichtlich aus der Bimodalität der Schlüpfkurven. Eine Methode zur Analyse und zum Vergleich der Kurven wird diskutiert. Das Diapauseverhalten, das sich R. nitidula erworben hat, kann als Methode betrachtet werden, mit der eine vorwiegend tropische Art in gemässigte Zonen vordringen konnte.