Role of uronic acids present in phytopathogenic fungi as inducers of polygalacturonases during autolysis

The presence of uronic acids in the culture fluid and mycelium of the fungi: Alternaria alternata, Botrytis cinerea, Drechslera halodes, Fusarium culmorum, Fusarium oxysporum, Monilinia fructigena, Mucor mucedo, Rhizopus stolonifer and Trichoderma hamatum was detected and quantified. In these fungi the concentration of uronic acids increased during the growth phase and the maximal concentrations were found at the end of the growth phase or onset of autolysis both in the mycelium as well as in the culture fluid. The uronic acids were metabolized during the first days of autolysis decreasing to constant levels until the end of the autolytic period studied.The variations in the activity of polygalacturonase and polymethylgalacturonase present in the culture fluid were determined at the onset and during autolysis in these fungi. These enzymic activities were found in the culture fluid of these fungi, with exception of M. rouxii, and they showed an increasing activity in the first days of autolysis and later a slight increase or decrease was observed. The presence of uronic acids in these phytopathogenic or saprophytic fungi and the low levels detected during autolysis could be related to the induction of pectic enzymes and the pathogenicity of these fungi.     

Relationship of the replication and incompatibility genes of an IncFVI haemolysin plasmid with other F-like haemolysin plasmids

The replication and incompatibility region of the IncFVI plasmid pSU502 has been isolated by in vitro DNA manipulation as part of a 12.6 kb plasmid, denominated pSU503. Plasmid pSU503 was strongly incompatible with its parental plasmid, pSU1, but was fully compatible with the haemolytic plasmids pSU316 (IncFIII/IV), pHly152 (IncI2) and pSU233 (Inc-pSU233). Furthermore, the 6.9 kb EcoRI fragment of pSU503 which carries the replication and incompatibility determinants of pSU1 did not show any detectable homology (less than 70%) with any of the haemolysin-determining plasmids with which it is compatible. Thus, homologous haemolysin determinants have become linked to apparently unrelated replicons.     

SYNTHESIS OF RNA IN NEURONS OF THE HYPOGLOSSAL NERVE NUCLEUS, AFTER SECTION OF THE AXON, IN MICE

Synthesis of RNA in neurons of the hypoglossal nerve nucleus after axonal section was studied by means of [5-³H]uridine administration and radioautographic counting techniques in mice. The results of the experiments were evaluated by counts of silver grains over the nucleoplasm and cytoplasm of the neurons. RNA synthesis was greater in neurons after axonal section, and this increase was evident from 12 hr after the operation. The greatest increases in the operated side were observed in the 1st, 2nd and 3rd days after operation. In the 7th and 14th days RNA synthesis was still greater in the hypoglossal nucleus of the sectioned nerve but the difference in the control nucleus was not so striking. In the 30th day synthesis of RNA in left and right hypoglossal nuclei was comparable.     

Chemical and physiological changes in fungi during autolysis

A comparative study was done on some of the chemical changes occurring during autolysis of cultures ofAspergillus flavus in both physiologically acid and alkaline media. The mycelium ofA. flavus lost during autolysis 44 % of its maximum dry weight in the physiologically alkaline medium, whereas this loss was apparently nil in the physiologically acid medium. Nitrogen containing compounds seemed not to be affected by autolysis either in the physiologically acid or alkaline media. The disappearance of P-containing compounds in mycelium ofA. flavus autolysed in both conditions (NO ₃ ^– and NH ₄ ⁺ as N source) amounted to 64 % in the alkaline autolysis and to nearly 77 % in the acid autolysis. The results we have obtained for the acid autolysis strongly suggest that very little activity is shown by autolytic enzymes in the interval 10–133 days of incubation, when measuring autolysis by the loss in mycelial dry weight.

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Zusammenfassung Eine vergleichende Untersuchung war unternommen an einigen der chemischen Veränderungen, die während der Autolyse der Kulturen vonAspergillus flavus in physiologischen sauren und alkalischen Medien vorkommen. Die Myzelien vonA. flavus haben während der Autolyse 44 % ihres größten Trockengewichtes in physiologisch alkalischem Medium verloren, während dieser Verlust in physiologisch saurem Medium anscheinend Null gewesen ist. Stickstoff enthaltende Substanzen erschienen während der Autolyse weder in physiologisch saueren noch in alkalischen Medien beeinflußt zu sein. Das Verschwinden von P-enthaltenden Substanzen in Myzelien vonA. flavus in Autolyse unter beiden Bedingungen (NO ₃ ^– und NH ₄ ⁺ als Stickstoffquelle) erreichte 64 % in alkalischer Autolyse und beinahe 77 % in der saueren Autolyse. Die Ergebnisse, die wir in der saueren Autolyse erhalten haben legen es sehr nahe, daß autolytische Enzyme eine sehr geringe Aktivität in der Zeitspanne von 10–133 Tagen der Inkubazion zeigen, wenn die Autolyse an dem Verlust des mycelialen Trockengewichtes gemessen wird.

     

Physiology of a choline-requiring strain of Torulopsis pintolopesii

Summary A choline-requiring strain of Torulopsis pintolopesii when growing on choline-methyl-¹⁴C or choline-methyl-³H excretes the radioactivity incorporated in the first 24–48 h of incubation up to ca. 85–95% of the radioactivity added at the beginning of the incubation. The addition of non-radioactive methionine did not interfere with the uptake and excretion of radioactivity from choline-Me-¹⁴C. Radioactive methyl group of methionine previously incorporated by growing cells of T. pintolopesii on varying concentrations of choline was not excreted in appreciable amounts. No evidence was obtained for the oxidation of choline to betaine, degradation to trimethylamine, or net incorporation of labelled choline into lecithin. The occurrence of a new pathway for the utilization of choline in yeasts is suggested. The requirement of choline by T. pintolopesii is explained tentatively by the formation and excretion of a compound containing the carbon and hydrogen atoms from the methyl groups of choline and whose chemical structure still under study, may comprise a heteroside containing mannitol as the polyhydroxylated moiety.     

Lesiones histoplasmóticas autópsicas como índice epidemiológico de la enfermedad en Los Andes Venezolanos

Resumen En 244 autopsias de los años 1961, 1962 y 1963 de sujetos de más de 2 años de edad fué encontrado un 25% de casos con lesiones histoplasmóticas, en su gran mayoría de carácter residual, en pulmones y ganglios linfáticos. Esta cifra corresponde exactamente al porcentaje de reactores positivos a la prueba de histoplasmina en la población de nuestra región. Fueron corroborados los resultados deStraub &Schwarz, quienes en Cincinnati/Ohio encontraron la misma concomitancia entre los resultados de estos métodos de estudio, ambos igualmente valiosos para determinar el indice epidemiológico de histoplasmosis en una zona determinada.Puesto que en nuestros estudios no fueron aplicadas las mismas técnicas completas como en Cincinnati las cifras positivas en nuestro ambiente pueden eventualmente ser más altas, pero no alcanzan las cifras aún más elevadas, reportadas de otras zonas de Venezuela. En nuestro material el número de casos con lesiones histoplasmóticas antiguas múltiples en el pulmón, que sugieren ser primarios, era mayor que en la primera serie deStraub &Schwarz. Parece que entre nosotros la infección tiene lugar en períodos más tempranos de la vida que en EEUU de Norteamérica.El más alto índice de infección, de casi 50%, fué encontrado en personas provenientes de la región de la Cuenca del Lago de Maracaibo, disminuyendo algo en los valles de baja altura de las montañas y llegando a cifras bajas en los lugares de mayor altura.

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Summary In 244 autopsies in patients from 2 years of age on, in 1961, 1962 and 1963 histoplasmic lesions were found in 25% of all cases, mostly residual foci in lungs and lymph nodes. This percentage is exactly the same as the percentage of histoplasmin sensitivity in our environment. Thus, the results ofStraub &Schwarz who found the same percentage of anatomic lesions and histoplasmin reactors in Cincinnati, Ohio, were confirmed in our material. Morphologic methods in autopsies are a valuable tool for epidemiologic investigations regarding histoplasmosis.Since a more exact technique of examination was applied in Cincinnati, it is probable that the positive percentage in our environment is even higher. A higher number of multiple residual histoplasmotic foci in the lungs was found in our series in comparison with the first material ofStraub &Schwarz. Multiple histoplasmic foci in some of these cases may be due to primary infection. Infection withH. capsulatum occurs apparently in our environment more frequently in younger persons.The highest infection rate of almost 50% was found in patients who lived in the tropical lowlands near the Lake of Maracaibo and the Panamerican Highway. A lower rate occurred in the mountain valleys up to approximately 3000 feet of altitude, and decreased even more in higher altitudes.

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Zusammenfassung In 244 Sektionen vom 2. Lebensjahr an aus den Jahren 1961, 1962 und 1963 wurde in 25% aller Fälle Histoplasmose in Form von Residualläsionen in Lungen und Lymphknoten gefunden. Diese Zahl entspricht genau dem Prozentsatz der Bevölkerung in unserer Gegend, die positiv auf Histoplasmin reagiert. Damit werden die Befunde vonStraub &Schwarz, die an Autopsien in Cincinnati, Ohio erhoben wurden, bestätigt. Neben der Histoplasminreaktion ist es auch mittels anatomischer Untersuchungen möglich, Aussagen über das Ausmass der Infektion mitHistoplasma capsulatum in einer bestimmten Gegend zu machen.Da bei den Untersuchungen in Cincinnati genauere Untersuchungsmethoden zur Anwendung kamen, ist es möglich, dass der Prozentsatz unserer positiven Fälle eher noch höher liegt. Die Zahl unserer Fälle mit multiplen residualen Histoplasmoseherden in den Lungen ist noch höher als in der ersten Untersuchungsserie vonStraub &Schwarz. Es könnte sich in einem Teil der Fälle durchaus um multiple Primärkomplexherde handeln. Anscheinend findet die Infektion mitH. capsulatum bei uns in früheren Lebensaltern statt.Die höchste Infektionsrate von fast 50% wurde in Personen aus den gegen den Maracaibosee zu gelegenen tropischen Gebieten gefunden. Sie war etwas geringer in den Gebirgstälern bis zu etwa 1000 m Höhe und gering in Gebieten grösserer Höhe.

     

Effect of lead on blood protoporphyrin levels of a group of ring teal ducks (Callonetta leucophrys)

This research determined the relationships between blood lead level and zinc protoporphyrin (ZnPROTO), protoporphyrin IX (PROTO), and free erythrocyte protoporphyrin (FEP) levels in a group of 18 ring teal ducks. Blood samples were drawn from two groups of ring teal ducks as part of the routine health maintenance program of the New York Zoological Park. One group of six teals had been exposed to what is believed to be lead-contaminated dust, while the second group of twelve teals was unexposed. Blood samples were analyzed for lead by flameless atomic absorption and for protoporphyrins by fluorescence. Blood lead level and log blood lead level had positive correlations with each of the protoporphyrins: the logarithmic correlations were better than the nonlogarithmic correlations, and PROTO correlated better than ZnPROTO. With one exception, PROTO levels were higher than ZnPROTO levels. The results suggest that PROTO, FEP, or ZnPROTO could serve as a biological indicator of lead poisoning in ring teal ducks.     

Role of recurrent hydrophobic residues in catalysis of helix formation by T cell-presented peptides in the presence of lipid vesicles

We tested the hypothesis that the recurrence of hydrophobic amino acids in a polypeptide at positions falling in an axial, hydrophobic strip if the sequence were coiled as an alpha helix, can lead to helical nucleation on a hydrophobic surface. The hydrophobic surface could anchor such residues, whereas the peptide sequence grows in a helical configuration that is stabilized by hydrogen bonds among carbonyl and amido NH groups along the peptidyl backbone of the helix, and by other intercycle interactions among amino acid side chains. Such bound, helical structures might protect peptides from proteases and/or facilitate transport to a MHC-containing compartment and thus be reflected in the selection of T cell-presented segments. Helical structure in a series of HPLC-purified peptides was estimated from circular dichroism measurements in: 1) 0.01 M phosphate buffer, pH 7.0, 2) that buffer with 45% trifluoroethanol (TFE), and 3) that buffer with di-O-hexadecyl phosphatidylcholine vesicles. By decreasing the dielectric constant of the buffer, TFE enhances intrapeptide interactions generally, whereas the lipid vesicles only provide a surface for hydrophobic interactions. The peptides varied in their strip-of-helix hydrophobicity indices (SOHHI; the mean Kyte-Doolittle hydrophobicities of residues in an axial strip of an alpha helix) and in proline content. Structural order for peptides with helical circular dichroism spectra was estimated as percentage helicity from circular dichroism theta 222 nm values and peptide concentration. A prototypic alpha helical peptide with three cycles plus two amino acids and an axial hydrophobic strip of four leucyl residues (SOHHI = 3.8) was disordered in phosphate buffer, 58% helical in that buffer with 48% TFE, and 36% helical in that buffer with vesicles. Percentage helicity in the presence of vesicles of the subset of peptides without proline followed their SOHHI values. Peptides with multiple prolyl residues had circular dichroism spectra with strong signals, but since they did not have altered spectra in the presence of vesicles relative to phosphate buffer alone, the hydrophobic surface of the vesicle did not appear to stabilize those structures.