Further environmental factors affecting the antigenicity of Trichophyton rubrum

Biochemical determinations performed on ammonium sulfate soluble and insoluble fractions of crude mycelial extracts ofTrichophyton rubrum indicated that these antigens were either carbohydrates or carbohydrates with peptide side chains. The antigens contained considerable amounts of galactose and mannose.Gel filtration techniques proved to be effective in separating these antigens. One antigen had a molecular weight greater than or equal to 2.0 × 10⁶. A smaller, more reactive antigen was also found; however, the elution time of this antigen varied with the concentration of dextrose in the medium.Quantitative precipitation tests used to differentiate the serological reactivity of crude antigens, recovered from mycelia grown on media containing variable concentrations of dextrose, indicated that the serological reactivity of the crude antigen was inversely proportional to the concentration of the carbohydrate source, with an optimum reactivity occurring with antigens prepared from mycelia grown in low dextrose concentrations.Nitrogen and carbohydrate concentrations performed on whole mycelia and cell free extracts demonstrated that the total nitrogen, soluble nitrogen, total carbohydrate and soluble carbohydrate concentrations were influenced by the concentration of the carbohydrate source. The optimum carbohydrate concentration necessary for the maximum ratio of protein and carbohydrates per gram of mycelium was 15.0 g/l. This is less than the amount used in most Sabouraud's dextrose media formulations. The effect of these environmental factors on the serological reactivity was discussed.Supported in part by NIH Environmental Health Tranining Grant ES00081-02. The help of Mrs. Gary Swecker is gratefully acknowledged for preparing the graphs.     

Cytologische Untersuchungen an Nematodengallen

Zusammenfassung Zwei verschiedene Faktoren bewirken die Vergrößerung der Riesenzellen (RZ) in den Gallen des NematodenMeloidogyne arenaria (auf Kakteen und anderen Wirten): die Hypertrophie der wachsenden RZ und die Syncytienbildung (Auflösung trennender Zellwände und Verschmelzung kleinerer Zellen).Parallel mit der Entwicklung des Parasiten durchlaufen die RZ und ihre Kerne vier verschiedene Entwicklungsstadien; währenddessen verändern diese Kerne auf charakteristische Weise ihre Größe, Struktur und Gestalt, parallel damit erhöht sich der Polyploidiegrad (die Charakteristika der einzelnen Stadien sind vom jeweiligen Wirt weitgehend unabhängig): der Umriß wandelt sich vorerst durch starke physiologische Beanspruchung des Kerns, in späteren Stadien durch davon unabhängige Mitosestörungen bzw. durch Spindel- und Plattenverschmelzungen während der synchronen Teilungen in den RZ (bei der CrassulaceeCotyledon treten Mikronuklei auf). Die beiden letztgenannten Vorgänge verursachen die Polyploidisierung sowohl in den RZ als auch in manchen unmittelbar an die RZ anschließenden parenchymatischen Zellen, während das übrige Gewebe weitgehend unbeeinflußt bleibt.Eng mit den genannten Ursachen hängt die sehr variable Zahl der Kerne pro RZ und ihre Struktur zusammen: im Stadium der größten physiologischen Beanspruchung der RZ ist der Kern sehr wolkig, später sind die Chromozentren sehr kompakt. Unabhängig vom jeweiligen Entwicklungsstadium der RZ ist das Chromatin an der Peripherie des Kerns konzentriert. Durch die Ursachen, die zu Polyploidisierung und variablem Umriß führen, kommt es zu wahrscheinlich plasmatischen Einfaltungen und Einschlüssen innerhalb des Kerns.Nicht nur im Gallen-, sondern auch im unbeeinflußten Gewebe zeigen Kerne ab einer bestimmten Größe bzw. eines bestimmten Polyploidiegrades stärker lichtbrechende, nicht oder nur wenig anfärbbare, in ihrer Größe zwar vom Kernvolumen abhängige, doch trotzdem kleine Kugeln (in kleineren Kernen sind sie wahrscheinlich nur wegen ihrer Kleinheit nicht auffindbar). Sie sind nur in Glutaraldehyd-fixiertem Material sichtbar, AE als Fixierungsmittel löst sie auf. Sie befinden sich oft in unmittelbarer Umgebung des Nukleolus und hängen wahrscheinlich ursächlich mit ihm zusammen, aber eine exakte Analyse kann nicht gegeben werden.

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Summary Two determining factors induce the enlargement of giant cells in galls caused by the root-knot nematode (Meloidogyne arenaria in roots of some Cactaceae and other hosts): hypertrophy of the growing giant cells and formation of syncytia.Corresponding with the evolution of the parasitic larva the giant cells and their nuclei become altered through four different stages; the nuclei change their volume, structure, shape and their degree of polyploidy, independent of the specific host: the contour of the nuclei is altered during the development of the giant cells first by physiological factors, on the other hand — later on — by mitotic inhibition resp. by fusing mitotic spindles or mitotic figures during synchronous mitotic divisions in the giant cells (micronuclei occur inCotyledon, Crassulaceae). Polyploidization is induced by the last two mentioned factors in giant cells as well as in some parenchymatous cells surrounding giant cells.Conditioned by these mentioned factors the number of nuclei per giant cell, their structure and shape are very variable. All nuclei in the giant cells possess a significant feature: accumulation of the chromatin material at the nuclear periphery, while the centre of the nucleus is almost optically empty. This structure occurs also during the stage with the greatest physiological stress. Plasmatical foldings and inclusions occur in some voluminous nuclei, produced by the factors leading to polyploidy resp. to variable shape.Not only in giant cells, but also in normal tissues — if their nuclei have reached a low degree of polyploidy — small, refractioning, poor stainable globules exist (they cannot be seen in small nuclei, probably they are too small): they are often sitting upon the nucleolus and are surely corresponding with him, their exact constitution and origin is unknown. They can only be seen in Glutaraldehyd-fixed material, in acetic-alcohol-fixation they are dissolved.

     

Antibacterial Action of Essential Oils of Artemisia as an Ecological Factor: II. Antibacterial Action of the Volatile Oils of Artemisia tridentata (Big Sagebrush) on Bacteria from the Rumen of Mule Deer

Rumen microorganisms of wild and captive deer were subjected to increasing amounts of volatile oils. The oils had a marked antibacterial effect on the rumen bacteria when the concentration reached approximately 16 muliters of oil per 10 ml of rumen fluid nutrient broth. The gross reactions of rumen bacteria obtained from wild, as well as captive, deer to the volatile oils seemed to be of the same magnitude; thus no adaptation by the bacteria to the oils was apparent.     

Secondary and Apparent Primary Antibody Responses After Group A Streptococcal Vaccination of 21 Children

A partially purified M protein, extracted from a mouse-virulent strain of type 3, group A streptococcus, was administered subcutaneously in gradually increasing amounts at weekly intervals to 21 children in a Family Program. Seven children with type 3 bactericidal antibody in prevaccination sera showed a secondary response. Of 14 children with no detectable type 3 bactericidal antibody prior to vaccination, 13 developed definite type 3 antibody during or soon after vaccination. This response appeared to be of the primary variety in at least some of the 13 children because (i) the total incidence of antibody response (20 of 21) was greater than can be accounted for by the documented incidence of clinical and subclinical type 3 infections among children of our Family Program during a period of 14 years, (ii) the response in the 13 children with no detectable antibody prior to vaccination was more delayed than in those showing a definite secondary response, and (iii) on the average, the amount of vaccine needed for a response in these 13 children was 15 to 28 times greater than that required for the secondary response. Local skin reactions were variable among the vaccinated children. Systemic reactions were infrequent and not severe. The giving of multiple injections of partially purified M protein did not seem to cause cutaneous hypersensitivity.     

Dry Heat or Gaseous Chemical Resistance of Bacillus subtilis var. niger Spores Included Within Water-soluble Crystals

Inclusion of spores of Bacillus subtilis var. niger in water-soluble crystals increased the resistance of the spores to dry heat and to a gaseous mixture of methyl bromide and ethylene oxide. Resistance of spores in glycine crystals to dry heat at 125 C was increased 5 to 24 times compared to unprotected spores. There appeared to be a positive correlation between the size of the crystal and the degree of resistance. The resistance to dry heat of spores included in sodium chloride crystals was about six times greater than unprotected spores. A gaseous mixture of methyl bromide (964 mg/liter) and ethylene oxide (642 mg/liter) at 37% relative humidity was ineffective in sterilizing spores enclosed within these water-soluble crystals, as was ethylene oxide alone. However, if the relative humidity was sufficiently high to dissolve the crystals during exposure to the vapor, viable-spore counts were drastically reduced or were negative. The surfaces of crystals grossly contaminated with dry spores were sterilized by exposure to gaseous ethylene oxide. Sterilization of heat-labile or moisture-labile materials with a critical requirement for sterility, as in planetary probes or drugs, may be complicated by the presence of spores in naturally occurring water-soluble crystals. This phenomenon is similar to the protection afforded spores entrapped in solid plastics.