Conformational changes in Sindbis virus envelope proteins accompanying exposure to low pH.

The attachment of high multiplicities of Sindbis virus to tissue-cultured cells followed by brief treatment at low pH has been shown to produce cell fusion (fusion from without). In this report, experiments to determine the effects of low pH on the physical and biological properties of Sindbis virus are described. Exposure of purified Sindbis virions to mildly acidic conditions resulted in a rapid and irreversible alteration in particle density and sedimentation characteristics, followed by a slower loss of infectivity. Infectivity was not restored by a return to neutral pH; rather, the loss of virus infectivity seemed to be initiated by exposure to low pH but continued at neutral pH. The formation of a virus-cell complex in which virions were attached to the cell surface protected the particles from low-pH inactivation, although low pH could still expose virus functions responsible for cell fusion. Low pH was found to induce a conformational change in the E2 polypeptide of the intact virion. These results are discussed with respect to the process of Sindbis virus infection of tissue-cultured cells.     

Structural alterations in fibronectin correlated with morphological changes in smooth muscle cells

Nontransformed cultures of vascular smooth muscle cells proliferate until they form a confluent sheet of cells. Subsequently, the cells become reorganized to form multicellular nodules that are loosely attached to the substrate. The formation of nodules is facilitated by the addition of medium conditioned by nodular cultures. Nodulation is inhibited by the addition of fibronectin. Fibronectins derived from monolayer culture conditioned medium or from plasma are maximally effective while fibronectin isolated from nodular cell conditioned medium is inactive. Analysis by NaDodSO4-polyacrylamide gel electrophoresis reveals that the nodular cell fibronectin has a molecular weight that is about 20-30 kd less than that of monolayer cell fibronectin. Further, nodular cell conditioned medium contains an activity that can convert both plasma fibronectin and monolayer cell fibronectin to the lower molecular weight correlated with the loss of biological activity.     

A paired-tracer dilution method for characterizing membrane transport in the perfused rat hindlimb. Effects of insulin, feeding and fasting on the kinetics of sugar transport.

We have applied the paired-tracer dilution method to the study of transport processes in a mixed mammalian muscle preparation, the perfused rat hindlimb. The method is suitable for the characterization of the kinetic parameters of sugar and amino acid transport and its regulation by hormones, contractile activity, hypoxia, etc. Insulin stimulates sugar transport by increasing the Vmax. of the process 2-3 fold, but its affinity is unchanged. Starvation increases the affinity of sugar transport in perfused skeletal muscle.     

Probing by aphids in the leaf tissues of resistant and susceptible sugar beet

The probing of Aphis fabae and Myzus persicae in the leaves of sugar beet with inherited resistance or susceptibility to aphids was studied by microscopic examination of samples of whole leaves, prepared after 48 h exposure to adult aphids at approximately three aphids cm^-2.The density of saliva stylet-sheaths left by the aphids (cm^-2) and the proportion reaching phloem differed between sugar beet stocks and were inversely associated. Differences in resistance between stocks could not, however, be related directly to either. All beet stocks examined were probed freely. Seasonal differences in sugar beet grown in the glasshouse affected the proportion of sheaths reaching the phloem, but the differences between beet stocks were similar at all times.The densities of sheaths left by different clones of M. persicae corresponded with the aphids' response to sugar beet as a host plant. Among aphid clones which readily colonize sugar beet, the densities of stylet sheaths which reached phloem suggested that the adults of both A. fabae and M. persicae gained sufficient access to sieve tubes to satisfy their nutritional needs. The phloem of sugar beet from the glasshouse was always within the estimated maximum depth to which the aphids probe; but, in leaves from the field, it appeared that the phloem might be inaccessible to young M. persicae in the sugar beet crop during late summer.

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Zusammenfassung Das Proben von Aphis fabae und Myzus persicae in Blättern von Zuckerrüben mit erblicher Blattlausresistenz bzw.-anfälligkeit wurde untersucht durch mikroskopische Durchmusterung von Speichelscheiden in Proben von ganzen Blatt. Rübenblätter wurden mit genähert drei adulten Läusen cm^-2 besetzt und nach 48 Stunden quergeschnittene Streifen der Blätter in Alkohol fixiert, gefärbt und mit der Unterseite nach oben auf Objektträgern eingeschlossen.23890 Speichelscheiden wurden registriert. Die Dichte der Scheiden von M. persicae (cm^-2) und der Anteil der das Phloem erreichenden Scheiden (SRP) unterschieden sich signifikant zwischen den Rübenstämmen. Bei A. fabae ergaben sich entsprechende, aber nicht gesicherte Unterschiede. Scheidendichte und Prozentsatz SRP waren gegenläufig, zwei Rübenstämme zeigten eine hohe Scheidendichte, zwei andere hatten weniger Scheiden, aber einen höheren Prozentsatz SRP. Diese Gruppierung der Stämme korrespondierte aber nicht mit ihrer Blattlausresistenz. Aus der Scheidendichte ergab sich, dass M. persicae und A. fabae auf allen geprüften Rübenstämmen, resistenten und anfälligen, unbehindert probten, so dass jede Laus das Phloem durchschnittlich etwa viermal am Tag erreichte. Ein Klon von M. persicae, der sich an Rüben nicht entwickelt, hinterliess weniger Scheiden in den Blättern aller Stämme.Der Anteil von SRP war bei Prüfungen im März grösser als im November. Dieser Unterschied war besonders deutlich bei Scheiden von Larven, die im übrigen zu allen Zeiten das Phloem weniger oft erreichten als ihre Eltern. Messungen des Abstandes von der unteren Blattfläche zum Phloem ergaben, dass das Phloem den Läusen in Gewächshaus-Zuckerrüben immer zugänglich war. M. persicae-Larven konnten jedoch in Blättern von Freilandrüben das Phloem nicht erreichen.

     

Fate of corticotrophins in an isolated adrenal-cell bioassay and decrease of peptide breakdown by cell purification

1. The fate of corticotrophins in a trypsin-dispersed rat adrenal-cell assay system was investigated with a view to establishing whether differences in the rate of inactivation might contribute to potency differences observed between analogues. 2. Corticotrophin-(1-24)-tetracosapeptide and to a lesser extent synthetic 1-39 corticotrophins were found to be inactivated during incubation with cell suspension. 3. Peptide fragments were isolated by using [[(3)H(2)]Tyr(23)]corticotrophin-(1-24)- tetracosapeptide as a marker. The fragments indicate a peptidase with a predominantly tryptic specificity. 4. The peptidase is present in the extracellular fluid and is released from cells when they are damaged. 5. Cells were fractionated on an albumin gradient. Cells from the zona fasciculata and the zona intermedia or reticularis were present in fractions which produced fluorogenic steroids in response to corticotrophin. 6. Purification of the cells by centrifugation through albumin decreased degradation by peptidases, so that if the assay is carried out with a dilute suspension of purified cells peptide breakdown should not affect the observed potencies of adrenocorticotrophin analogues. 7. No binding of [[(3)H(2)]Tyr(23)]corticotrophin-(1-24)- tetracosapeptide to cells could be detected at low concentrations of the peptide. This indicated that less than 120 receptors/cell are occupied during stimulation by a dose that would elicit approx. 80% of the maximal response.     

Influence of the Maize Dwarf Mosaic Virns and Simazine upon the Buffer Soluble Proteins and Catalase Activity of a Virus-susceptible and a Virus-tolerant Variety of Zea mays

Inbred corn varieties differing in their tolerance to the maize dwarf mosaic virus infection also differ in varietal character and host response to simazine. Oh 7B, a tolerant cv., has less total soluble and globulin proteins than the sensitive variety, H 55, but more Kjeldahl nitrogen, albumin protein and a doubling of catalase activity. Virus inoculation in Oh 7B causes a reduction in all three protein fractions, Kjeldahl nitrogen and catalase activity. Deleterious virus-simazine interaction does not occur with either variety and the observed varietal and host response differences in protein may account for the conflicting reports of other workers.