Membranes of Saccharomyces cerevisiae

A crude small particle pellet, obtained from postmitochondrial supernatant fractions of Saccharomyces cerevisiae, contains about half the ergosterol and phospholipid of crude cell homogenates. Most of the phospholipid of this pellet is in a “heavy” fraction which, with the aid of electron microscopy, shows membranous elements in addition to discrete particles. The “heavy” fraction, upon treatment with deoxycholate, can be freed of membranes, or, with ribonuclease treatment, ribosomes can be removed, leaving relatively clean membranes. The “heavy” fraction resembles the microsomes of animal cells, but contains considerably less lipids, including phospholipids, thus suggesting a less well-developed intracellular membrane system.     

Temperature Response in Animals Infected with Bacillus anthracis

Rats, rabbits, swine, guinea pigs, and monkeys were infected with anthrax spores, and their temperature responses were recorded. These were characteristic for a species and appeared independent of resistance or susceptibility of the species toward establishment of the disease. The rabbit appeared unique in that it not only failed to demonstrate a dose-response relationship over an 8-log dose range, but acted independently producing erratic body temperatures depending on spore dose. This limits the usefulness of the rabbit in studying anthrax pathogenesis, and poses questions regarding published data with the rabbit as the test animal.     

Spectrophotometric Measurements of Phytochrome in vivo and Their Correlation with Photomorphogenic Responses of Phaseolus

Direct in vivo measurements of phytochrome have been made in Phaseolus vulgaris by 2-filter difference spectrophotometry (Ratiospect). All measurements were made at 730 versus 800 nm and it is assumed that the Δ (ΔOD) is directly proportional to the PFR concentration of phytochrome present. Dose response curves were determined for both physiological and spectrophotometric responses for red induction and far-red photoinactivation. For induction, saturation occurs at 100 mj/cm² and for inactivation at 30 mj/cm². The rate of hook opening and the physiological response measured 20 hours after induction are both shown to be directly proportional to the initial amount of PFR present spectrophotometrically. The sensitivity of the tissue correlates well with the absolute amount of phytochrome present, the inner portion of the hook having the maximum concentration of 0.042 Δ (ΔOD)/g fresh weight. If the total reversible phytochrome concentration is reduced by exposure to red light and allowing PFR to decay out of the system the remaining sensitivity of the tissue is shown to be directly correlated with the amount of PR remaining in the tissue. PFR disappears rapidly in the dark at 25°, and is not detectable after 6 hours. There is no indication that PFR reverts in the system to PR. At 4°, PFR does not disappear measurably up to 1 hour and is nearly totally reversible to PR.     

On the ancestral form of true organ of fructification in the saprophytic stage of the dermatophytes of the faviform group: Favomicrosporon pinettii,N.SP. and its perfect form: Anixiopsis stercoraria (hansen) hansen, 1897

Summary The discovery of the hidden, built-in macroconidia in the four members of the Faviform Group of the dermatophytes, i.e.,Achorion schoenleinii, Trichophyton violaceum, Trichophyton verrucosum andMicrosporon ferrugineum, is described.To bring the hidden, built-in macroconidia to full fructification, i.e., to force the production of imperfect and perfect organs of fructification (macroconidia, cleistothecia), two entirely different techniques have been used: 1) the hair-soil method, 2) the yeast extract method.The two techniques, entirely independent from each other, yielded the same result: the ancestral form of the four members of the Faviform Group of dermatophytes. The imperfect form is described asFavomicrosporon pinettii,Benedek, 1965, sp. nov. The perfect form isAnixiopsis stercoraria (Hansen)Hansen, 1897.The ancestral form was found not only in and cultured from the strains of those dermatophytes derived from pathological material, but it was also recovered from its saprophytic habitat, from the soil (potting soil).

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Zusammenfassung Die Entdeckung der verborgenen, eingebauten Makrokonidien in den vier Representanten der Faviformen Gruppe der Dermatophyten, i.e.Achorion schoenleinii, T. violaceum, T. verrucosum, Microsporon ferrugineum, wird beschrieben.Um die verborgenen, eingebauten Makrokonidien zur vollen Fruchtbildung zu bringen, i.e. um die Produktion der imperfekten und perfekten Organe der Fruktifikation (Makrokonidien, Kleistothecien) zu erzwingen, sind zwei völlig verschiedene Methoden benutzt worden: 1) die Haar-Erde-Methode, und 2) die Hefeextrakt-Methode.Beide Methoden, völlig unabhängig von einander, haben zu demselben Ergebnis geführt, i.e. zur Entdeckung der Urform von den vier Representanten der Faviformen Gruppe der Dermatophyten. Die imperfekte Form wird alsFavomicrosporon pinettii,Benedek, 1965, sp. nov. beschrieben. Die perfekte Form istAnixiopsis stercoraria (Hansen)Hansen, 1897.Die ancestrale Form wurde nicht nur aus den Stämmen jener Dermatophyten gezüchtet, die aus pathologischen Produkten gewonnen worden sind, sondern auch aus dem natürlichen Habitat: von der Erde (potting soil).

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Résumé La découverte des macroconidies occultes et encastrées dans les quatre membres du Groupe Faviforme des dermatophytes:Achorion schoenleinii, Trichophyton violaceum, Trichophyton verrucosum etMicrosporon ferrugineum, est décrite.Pour forcer les macroconidies occultes et encastrées à la fructification complète, i.e. de produire des organs de reproduction imparfaits et parfaits, macroconidies et cleistothecia, on a fait l'usage de deux techniques complètement différentes: 1) des cheveux sur sol et 2) de l'extraction de levure.Toutes les deux méthodes, complètement indépendantes l'une de l'autre, ont produit le même résultat: la forme ancestrale des quatre membres de la Groupe Faviforme des dermatophytes. La forme imparfaite est décrite commeFavomicrosporon pinettii,Benedek, 1965, sp. nov. et la forme parfaite commeAnixiopsis stercoraria (Hansen)Hansen, 1897.La forme ancestrale a été trouvée non seulement dans les souches des dermatophytes indiquées et en cultivées, provenantes des produits pathologiques, mais aussi du sol, du terrain jardinier.

     

Single-step purification of pertussis toxin and its subunits by heat-treated fetuin-sepharose affinity chromatography

A general procedure for purifying biologically active pertussis toxin from Bordetella pertussis fermentation broth using affinity chromatography on heat-treated fetuin-Sepharose CL-4B is described. Diethanolamine is used as eluent in this single-step purification to prepare endotoxin-free pertussis toxin in good yield (70%) and high purity (greater than 95%). This one-step affinity chromatography procedure can be easily applied for large-scale preparation of pertussis toxin S1 subunit and its B-component. The affinity-purified S1 subunit is devoid of any of the histamine-sensitizing activity normally associated with pertussis toxin. The chromatographically purified pertussis toxin and its subunits retained their immunogenicity and could induce high levels of anti-toxin neutralizing antibodies.     

Resistance to H-2-restricted but not to allo-H2-specific graft and cytotoxic T lymphocyte responses in lymphoma mutant

The lymphoma mutant RMA-S escaped graft rejection after transplantation over a minor histocompatibility barrier, whereas it was rejected in H-2 allogeneic mice. The parental control line was rejected in both situations. The mutant, which had been selected against MHC class I molecules retained 5 to 10% of the wild-type H-2Db, Kb, and beta 2-microglobulin expression on the cell surface. It remained sensitive to allo-H-2b CTL in vitro, but was completely resistant to minor histocompatibility antigen-specific, H-2b-restricted CTL. It was equally resistant to other H-2b-restricted responses against internally derived Ag, such as tumor-specific CTL or a CTL clone specific for the influenza virus nucleoprotein. The results indicate a target cell defect that selectively abolishes the sensitivity to H-2-restricted CTL directed against internally processed Ag. This appears sufficient to shift the transplantation response over a minor histocompatibility Ag barrier from rejection to acceptance. There are two possible explanations for the results: 1) a block in the MHC class I-directed pathway for internal Ag processing, and 2) subthreshold H-2/Ag ligand density in relation to triggering requirements of restricted CTL. Regardless of the type of defect, the results demonstrate a difference between allo-H-2-specific and H-2-restricted CTL recognition at the level of the target cell.     

A rapid posttranslational myristylation of a 68-kD protein in D. discoideum

Cells incubated with [3H]myristate were shown to rapidly and specifically acylate a 68-kD protein, p68, in a developmentally-regulated manner. The fatty acid incorporated into p68 was identified as myristate, and is linked to the protein via an amide bond, apparently to an NH2-terminal glycine. The acylation of p68 in D. discoideum displays some unusual properties. Unexpectedly, myristylation of p68 is a posttranslational event and occurs in the presence of inhibitors of protein synthesis. Another unusual finding was that although p68 is a stable protein, the acyl moiety is removed with a half time of approximately 15 min.