Exposure of mice to inhalation of pigmented conidia of Sporothrix schenckii

Summary We have tried to infect mice by inhalation with pigmented conidia ofSporothrix schenckii, in an apparatus ofPiggott &Emmons using pieces of wood artificially contaminated in the laboratory.In a first experiment the infected pieces of wood were placed in the bottom of the flask. We could not infect any of the exposed animals. In the second experiment the pieces of wood were suspended inside the flask and moreover, calcium chloride crystals were used to avoid humidity. A much higher liberation of spores was observed as comparing with the first experience. In two out of the 20 animals positive cultures ofS. schenckii were obtained from the lungs of one and from the liver and spleen of the other.We infer the difficulty in infecting mice with pigmented conidia of the pathogen by this procedure. It is presumed that these results agree with the rarity of the pulmonary forms in sporotrichosis.

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Resumen Se intentó la infección de lauchas mediante inhalación en aparato dePiggott &Emmons, con conidios pigmentados deSporothrix schenckii partiendo de maderas artificialmente contaminadas en el laboratorio. En una primera experiencia las maderas infectadas fueron colocadas en el fondo del aparato. No se logró infectar ninguno de los 14 animales inhalados.En una segunda experiencia las maderas fueron colocadas suspendidas dentro del matraz y además se introdujeron cristales de cloruro de calcio para disminuír la cantidad de vapor de agua. Se comprobó un mayor desprendimiento de esporos con relación a la experiencia preliminar y en dos de los 20 animales inhalados se logró cultivarS. schenckii a partir de pulmón en uno y de higado y bazo en otro. Se deduce la dificultad de infectar lauchas con conidios pigmentados del hongo por este procedimiento.Estos resultados concuerdan con la rareza de la puerta de entrada pulmonar en la esporotricosis.

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Departamento de Parasitología (Sección Micología). Instituto de Higiene, Montevideo, Uruguay.     

Effects of Na+, H+, and Cl− on alanine transport by lobster hepatopancreatic brush border membrane vesicles

Summary Epithelial brush border membrane vesicles (BBMV) of lobster hepatopancreas were formed by a magnesium precipitation technique previously described (Ahearn et al. 1985).³H-l-alanine transport by these vesicles was sodium and potassium insensitive, in contrast to a strong Na-dependency exhibited by³H-d-glucose transport. Initial alanine entry rates (15 s uptake) were stimulated and transient alanine uptake overshoots were observed when external pH was acidic (e. g. pH 4.0, 5.0 or 6.0) and a Cl gradient was imposed across the vesicular wall; at pH_o=7.4 alanine uptake was reduced in rate and hyperbolic in character. Alanine uptake from an acidic extravesicular environment in the absence of Cl responded to a transmembrane electrical potential difference created by an outwardly-directed, valinomycin-induced, potassium diffusion potential, suggesting that the alanine molecule alone carried sufficient charge under these conditions to respond to the electrical gradient. External 5.0 mMl-lysine andl-serine similarly inhibited the influx and overshoot properties of 0.05 mM³H-l-alanine uptake, whereas 5.0 mMl-leucine had virtually no effect. Trans-stimulation of alanine initial uptake rates and an enhancement of alanine accumulation against a concentration gradient were observed by vesicles preloaded with 1 mMl-lysine, but not by vesicles lacking amino acids or those containing 1 mMl-leucine orl-serine.³H-l-alanine influx from acidic external environments in the presence of a Cl gradient occurred by a combination of carrier-mediated transfer and apparent diffusion. Decreasing pH_o from 6.0 to 4.0 elevated alanineK _t from 0.55 to 2.64 mM, while alanineJ _M increased from 55 to 550 pmol/mg protein· 15 s. Apparent diffusional permeability of the membranes to alanine under these conditions increased slightly. These results suggest, but do not conclusively prove, that alanine transport across BBMV of lobster hepatopancreas may occur by way of a classical y+ transprot protein at acidic pH. The extent of this transport is determined by the magnitude of the transmembrane chloride gradient which serves as a powerful driving force for cationic amino acids in this tissue.     

Factores Determinantes de la Fase Levaduriforme del Paracoccidioides brasiliensis

Resumen Se cultivaron 4 cepas autóctonas deP. brasiliensis en los medios líquidos siguientes: 1) medio con neopetona Difco dialisado deEvans &Kessel; 2) caldo infusión de carne glucosado al 1 %; 3) caldo extracto de carne glucosado al 1 % y 4) medio de Sabouraud glucosado al 4 %, distribuidos en frascos de Erlenmeyer de 300 cc a razón de 150 ml cada uno e incubados a 28°C en un agitador mecánico de vaivén con un desplazamiento de 5 cm y a 90 ciclos por minuto. En estas condiciones se obtuvo, en todas las cepas, la transformación de la fase filamentosa en levaduriforme o parasitaria con producción de abundantes microformas de 1 µ, aproximadamente, de diámetro.

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Summary We have cultured 4 autochthonous strains ofP. brasiliensis in the following liquid media: 1) media with dialysed Difco Neopeptone ofEvans &Kessel; 2) 1 % glucose meat infusion broth; 3) 1 % glucose meat extract broth and 4) 4 % glucose Sabouraud media, distributed in 300 cc Erlenmeyer flasks containing 150 ml each, incubated at 28°C in an electric shaker with a speed range of 90 strokes per minute and a wide degree of agitation of 2'.We obtained under these conditions the conversion of the filamentous phase of the four strains into the yeast phase producing abundant microforms of about 1 µ of diameter.

     

Engineering of sugar metabolism of Corynebacterium glutamicum for production of amino acid l-alanine under oxygen deprivation

Corynebacterium glutamicum was genetically engineered to produce l-alanine from sugar under oxygen deprivation. The genes associated with production of organic acids in C. glutamicum were inactivated and the alanine dehydrogenase gene (alaD) from Lysinibacillus sphaericus was overexpressed to direct carbon flux from organic acids to alanine. Although the alaD-expressing strain produced alanine from glucose under oxygen deprivation, its productivity was relatively low due to retarded glucose consumption. Homologous overexpression of the gapA gene encoding glyceraldehyde 3-phosphate dehydrogenase (GAPDH) in the alaD-expressing strain stimulated glucose consumption and consequently improved alanine productivity. In contrast gapA overexpression did not affect glucose consumption under aerobic conditions, indicating that oxygen deprivation engendered inefficient regeneration of NAD⁺ resulting in impaired GAPDH activity and reduced glucose consumption in the alanine-producing strains. Inactivation of the alanine racemase gene allowed production of l-alanine with optical purity greater than 99.5%. The resulting strain produced 98 g l⁻¹ of l-alanine after 32 h in mineral salts medium. Our results show promise for amino acid production under oxygen deprivation.     

Biostatistical studies on western EuropeanDactylorhiza (Orchidaceae)—theD. maculata group

Multivariate analysis tools are exploited on a data set composed of quantitative characteristics collected on 35 populations of plants of theDactylorhiza maculata (L.)Soó group from Western-Europe. These samples lead to four well-defined clusters; this, together with qualitative, cytological and ecological arguments, allows for the recognition of four specific entities:D. maculata s.str.,D. fuchsii (Druce)Soó,D. saccifera (Brongn.)Soó andD. caramulensis (Vermeulen)Tyteca. It is concluded that the floral characters play an essential role in the taxonomical distinction. It also appears that the set of characters measured, as well as the methods exploited, are especially well-suited and valuable tools for the morphological study of the genusDactylorhiza.     

Some properties of phenylethanolamine-N-methyltransferase of rat brain

Phenylethanolamine-N-methyltransferase (PNMT, EC 2.1.1.28) was partially purified from rat brain. Brain homogenates were subjected to ultracentrifugation, salt fractionation, and gel filtration on Sephadex G-100. To compare the rat brain PNMT with that of adrenals, the same procedure was carried out with rat adrenal homogenates. The brain enzyme was eluted from Sephadex as a single fraction with a molecular weight of 26,900, while the enzyme from adrenals under the same conditions appeared in two fractions with molecular weights of 38,700 and 108,500. The brain fraction separated on Sephadex G-100 was active on phenylethanolamine substrates and inactive on indoleamine and phenylethylamine substrates. Products of the enzyme reaction were identified by bidimensional thin-layer chromatography asN-methyl derivatives of the corresponding amines. Kinetic studies showed that the type of inhibition of PNMT from rat brain and rat adrenals by SK&F 7698 was the same as described for PNMT from rabbit adrenals. Also, when normetanephrine andS-adenosyl-l-methionine were used as substrates, the apparentK _m values found with PNMT from rat adrenals and rat brain were similar.Preliminary reports were presented at XXV Convención Anual AsoVAC, Caracas, Venezuela, October 1975, and at XII Congreso Latinoamericano de Ciencias Fisiológicas, Bogotá, Colombia, November 1975.     

Production of <Emphasis Type="SmallCaps">l</Emphasis>-alanine by metabolically engineered <Emphasis Type="Italic">Escherichia coli</Emphasis>

Escherichia coli W was genetically engineered to produce l-alanine as the primary fermentation product from sugars by replacing the native d-lactate dehydrogenase of E. coli SZ194 with alanine dehydrogenase from Geobacillus stearothermophilus. As a result, the heterologous alanine dehydrogenase gene was integrated under the regulation of the native d-lactate dehydrogenase (ldhA) promoter. This homologous promoter is growth-regulated and provides high levels of expression during anaerobic fermentation. Strain XZ111 accumulated alanine as the primary product during glucose fermentation. The methylglyoxal synthase gene (mgsA) was deleted to eliminate low levels of lactate and improve growth, and the catabolic alanine racemase gene (dadX) was deleted to minimize conversion of l-alanine to d-alanine. In these strains, reduced nicotinamide adenine dinucleotide oxidation during alanine biosynthesis is obligately linked to adenosine triphosphate production and cell growth. This linkage provided a basis for metabolic evolution where selection for improvements in growth coselected for increased glycolytic flux and alanine production. The resulting strain, XZ132, produced 1,279 mmol alanine from 120 g l⁻¹ glucose within 48 h during batch fermentation in the mineral salts medium. The alanine yield was 95% on a weight basis (g g⁻¹ glucose) with a chiral purity greater than 99.5% l-alanine. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

habitat de Nocardia asteroides,Phialophora pedrosoi y Cryptococcus neoformans en Venezuela

Resumen Se estudiaron 336 muestras de tierra, madera y plantas provenientes de diferentes zonas del Edo. Mérida/Venezuela y Estados vecinos. Fueron inoculados cobayos y hamsters intratesticularmente y ratones por via endovenosa. Testículos y pulmones respectivamente fueron examinados por medio de cultivos e histológicamente.De 11 muestras se aislaronNocardia asteroides, de 6Phialophora pedrosoi y de unaCryptococcus neoformans.Se discuten las probables causas de porqué no se logró aislarParacoccidioides brasiliensis.

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Summary 336 samples of soil, wood and plants from several aereas of the State of Mérida/Venezuela and adjacent regions were studied.The samples were injected into the testes of guinea pigs and into mice intravenously. The testes and lungs of the animals respectively were cultured and examined histologically. From eleven samples were isolatedNocardia asteroides, from sixPhialophora pedrosoi and from oneCryptococcus neoformans.It is discussed, why it was not possible to isolateParacoccidioides brasiliensis.

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Zusammenfassung Es wurden 336 Erd-, Holz- und Pflanzenproben aus verschiedenen Gebieten des Staates Mérida/Venezuela und benachbarten Zonen untersucht. Das Material wurde Meerschweinchen intratestikulär und weissen Mäusen intravenös injiziert. Hoden und Lungen der Tiere wurden auf Nährböden verbracht und histologisch untersucht.Elfmal wurdeNocardia asteroides, sechsmalPhialophora pedrosoi und einmalCryptococcus neoformans isoliert und histologisch nachgewiesen. Es werden die möglichen Gründe diskutiert, warum es nicht gelangParacoccidioides brasiliensis zu isolieren.

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Del Instituto de Anatomía Patológica y Laboratorio de Micología de la Universidad de Los Andes, Mérida/Venezuela y del Laboratorio de Micología, Escuela de Medicina de la Universidad de Cincinnati/Ohio EEUU.

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Este trabajo pudo realizarse con la ayuda económica proporcionada por el Sr. Rector de la U.L.A., Dr.P. Rincón Gutiérrez a quien damos nuestras gracias más sinceras.A Wm. Bridge Cooke, Ph. D. y al Dr.G. A. de Vries agradecemos la identificación de las cepas dePhialophora pedrosoi.

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A los Dres.R. Kohler yP. N. Tablante G., y al Sr.Oswaldo Juergenson nuestro agradecimiento por la ayuda prestada en la recolección de las muestras.     

ACandida maltosa mutant defective in alanine aminotransferase: isolation andl-alanine assimilation

Candida maltosa JCM1504 can grow well onl-alanine as a sole carbon and nitrogen source. We found that the activities of alanine aminotransferase (AlaAT) and NAD-dependent glutamate dehydrogenase were remarkably induced when glucose-grown cells were transferred to medium containingl-alanine. This suggested thatC. maltosa has an induciblel-alanine degradation system including the above two enzymes. To assess whether AlaAT is essential for the first step ofl-alanine degradation, we isolated mutant N-07, which was unable to usel-alanine as a nitrogen source, from the wild strain. Mutant N-07 was very similar to the wild strain in terms of growth on pyruvate and on various amino acids other thanl-alanine, suggesting that N-07 lacked onlyl-alanine-assimilating ability. The AlaAT activity in the cell extract of N-07 was very low and was not induced byl-alanine, whereas the NAD-dependent glutamate dehydrogenase activity was the same as that of the wild strain and was inducible. Western blots with antibody raised against purified AlaAT fromC. maltosa indicated that no AlaAT protein was expressed in the mutant N-07. The low level of AlaAT activity described above was possibly due to the pyruvate-forming activity of other enzymes under the assay conditions. From these results, we concluded that AlaAT is an indispensable key enzyme forl-alanine assimilation inC. maltosa.     

The effects of adenohypophysectomy on the hypothalamic-hypophysial neurosecretory system and the adrenal gland of the toad Bufo arenarum Hensel

Summary The effects of adenohypophysectomy were studied on the hypothalamichypophysial neurosecretory system and the adrenal glands of Bufo arenarum Hensel. An increase in vascularization of the pars intermedia was found and the neurosecretory material (NSM) in the glandular region of the median eminence disappeared. Its reappearance later was accompanied by hypertrophy of the pars intermedia; differentiated chromophil cells appeared in the pars intermedia around the vessels and, at the same time, the adrenal reverted to normal. These findings are interpreted as hypertrophy and differentiation of the pars intermedia with replacement, at least partly, of the functions of the pars distalis. The probable mechanism of differentiation and the functions of chromophil cells of the pars intermedia are discussed.This paper was presented at the VI Reunión de la Asociación Latinoamericana de Ciencias Fisiológicas (A.L.A.C.F.), Viña del Mar, Chile, 1964. It was carried out under the auspices of the Consejo Nacional de Investigaciones Científicas y Técnicas (Argentina) and the Rockefeller Foundation (School grant RF-58028).Fellow of the Consejo Nacional de Investigaciones Científicas y Técnicas, Argentina.The authors wish to thank Prof. M. H. Burgos for his constant interest, Prof. B. A. Houssay, Prof. H. Heller and Dr. J. H. Tramezzani for their criticisms, Miss B. Rodriguez and Mr. L. Castro for their technical assistance.     

Effect of ammonium nitrate on nodulation and nitrogen fixation (acetylene reduction) of the tropical legumeSesbania rostrata

Summary The effect of ammonium nitrate on nodulation and nitrogen fixation (acetylene reduction activity) was investigated usingSesbania rostrata, a tropical legume with the ability to nodulate both roots and stems. Plants were grown in plastic pots on silica sand and gravel, inoculated one month after sowing and then continuously irrigated with an automatic irrigation system. Four nitrogen treatments were applied: 0, 1.5, 3.0 and 6.0mm NH₄NO₃. Related symbiotic parameters were evaluated 20 days later. With 3.0mm NH₄NO₃, root nodulation was inhibited. At that concentration, stem nodulation was not affected, but related nitrogenase activity decreased 85% and was completely inhibited at 6.0mm NH₄NO₃. Increasing NH₄NO₃ concentration resulted in a diminution of stem nodule size. Stem and root acetylene reduction were strongly inhibited by high mineral nitrogen concentrations; however root nitrogen fixation was more affected than stem nitrogen fixation.

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Resumen Se estudió el efecto del nitrato amónico en la nodulación y la fijación de nitrógeno, medida mediante la reducción de acetileno, utilizandoSesbania rostrata, una leguminosa tropical que forma nódulos tanto en raíz como en tallo. Las plantas se cultivaron en arena de sílice y grava, en macetas de plástico, se inocularon un mes después de la siembra y a partir de este momento se regaron continua y automáticamente. Se aplicaron cuatro tratamientos de N; 0, 1.5, 3.0, y 6.0mm NH₄NO₃ Los parámetros relativos a la simbiosis se evaluaron 20 días después. La nodulación en raíz se inhibió con 3.0mm NH₄NO₃, sin embargo a esta concentración la nodulación en tallo no estaba inhibida aunque la actividad nitrogenasa disminuyó en un 85%, quedando totalmente inhibida a 6mm NH₄NO₃. El incremento en la concentración de nitrato amónico produjo una disminución del tamaño de los nódulos del tallo. Elevadas concentraciones de nitrógeno mineral inhibieron fuertemente la reducción de acetileno tanto a nivel de raíz como de tallo, aunque la fijación de nitrógeno resultó más afectada en la raíz.

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Résumé L'effet du nitrate d'ammonium sur la nodulation et la fixation d'azote (réduction de l'acétylène) a été étudié avec la légumineuse tropicaleSesbania rostrata, qui présente la particularité de noduler à la fois sur la tige et sur les racines. Les plantes ont été cultivées dans des pots en plastique sur un mélange constitué de sable siliceux et de gravier. Un mois après le semis, elles ont été inoculées et placées sous irrigation continue au moyen d'un système automatique permettant l'apport du milieu nutritif. Quatre concentrations en azote ont été utilisées: 0, 1,5, 3,0 et 6,0 mmol/l NH₄NO₃. Les paramètres symbiotiques ont été analysés 20 jours plus tard. La nodulation racinaire est totalement inhibée en présence de 3,0 mmol/l NH₄NO₃. Cette concentration en azote combiné n'a pas d'effet apparent sur la formation des nodules de tige, mais leur activité nitrogénase diminue de 85%; leur activité est totalement inhibée en présence de 6,0 mmol/l NH₄NO₃. L'augmentation de la concentration en nitrate d'ammonium, se traduit par une diminution de la taille des nodules de tige. Les fortes concentrations en azote combiné ont un effet inhibiteur très marqué sur les activités réductrices d'acétylène racinaire ou caulinaire deS. rostrata; toutefois cet effet est plus sensible au niveau racinaire qu'au niveau caulinaire.

     

Histoplasma capsulatum en el suelo de nuestro ambiente. I. Examenes micologicos

Resumen En 156 muestras de suelo del Edo. Médira/Venezuela y de zonas vecinas se encontróHistoplasma capsulatum en 13(8,3%) mediante la técnica deEmmons modificada porGeorg, Ajello &Papageorge ySilberg &Hinton.Dos muestras positivas provenian de cuevas, una en el Edo. Mérida, otra en el Edo. Zulia, al sur de Machiques.Las 11 muestras restantes eran preferentemente del piso de gallineros descubiertos de 4 poblaciones en la vertiente occidental de Los Andes venezolanos, al sur del Lago de Maracaibo.Mientras que se pudo determinar de ésta manera el habitat del hongo en la zona cerca de la carretera Panamericana, resultaron negativas todas las muestras de las zonas más altas del Edo. Mérida provenientes preferentemente de lugares a lo largo de la carretera Transandina.Se hacen algunos comentarios acerca de la epidemiología de la histoplasmosis comparando los resultados obtenidos en ésta encuesta con la procedencia de los sujetos con histoplasmosis en el material autópsico y biópsico y los resultados de las pruebas de histoplasmina en la población de nuestra región.Además de los cultivos del hígado y bazo de los ratones se realizaron concomitantemente exámenes histológicos.En otro lugar se dará a conocer los resultados de éstos estudios comparandolos con los hallazgos micológicos.

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Summary Histoplasma capsulatum was found in 13 (8,3%) out of 156 samples of soil from the State of Mérida (Venezuela) and adjacent zones by means of theEmmons technique modified byGeorg, Ajello &Papageorge andSilberg &Hinton.Two positive samples had been taken from caves located in the States of Mérida and Zulia (south of Machiques), respectively.The remaining eleven (positive) samples came predominantly from the soil of chicken yards found in four villages situated on the western slopes of the Venezuelan Andes south of Lake Maracaibo.Whilst it was possible in this way to determine the habitat of the fungus in the zone near the Pan-American Highway, all the samples from the higher zones of the State of Mérida — coming predominantly from sites along the Trans-Andian Highway — proved to be negative.Some comments are made on the epidemiology of histoplasmosis, comparing the results obtained in this investigation with the origin of the subjects showing signs of histoplasmosis in the autopsic and biopsic material and the results of the histoplasmine tests carried out amongst the population of our region.Apart from the cultivation of rat livers and spleens, histological examinations were made concomitantly. The results of this research work as well as a comparison with mycological findings will be published elsewhere.

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Zusammenfassung In 156 Bodenproben aus dem Staat Mérida/Venezuela und angrenzenden Gebieten fand sichHistoplasma capsulatum in 13 (8,3%). Die Proben wurden nach den Angaben vonEmmons, die später vonGeorg, Ajello &Papageorge undSilberg &Hinton modifiziert wurden, untersucht.2 positive Proben stammten aus je einer Höhle in den Staaten Mérida und Zulia.Die restlichen positiven Bodenproben stammten hauptsächlich aus unbedeckten Hühnerställen von 4 Dörfern am Westabhang der venezolanischen Anden, im Süden des Maracaibosees.Während der Habitat der Pilze auf diese Weise in der Gegend der panamerikanischen Autostrasse festgestellt werden konnte, waren alle Proben aus den hochgelegenen Gebrigszonen negativ.Es wird die Frage aufgeworfen, wo und wie sich unsere Patienten mit nachgewiesener Histoplasmose infiziert haben und wie die Histoplasminsensibilität der Bevölkerung in den Gebirgsgegenden zu erklären ist.Ausser den Kulturen von Mäuselebern und -Milzen wurden histologische Untersuchungen durchgeführt, über die an anderer Stelle berichtet werden soll.

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El doctorCapretti, Profesor de Micología de la Universidad de Los Andes, falleció en el mes de diciembre de 1960 a consecuencia de una infección pulmonar porHistoplasma capsulatum contraida durante la ejecución de éste trabajo.Publicamos los resultados obtenidos de éstos estudios en homenaje póstumo a nuestro querido amigo, colaborador y maestro.     

Response ofWestiellopsis prolifica andAnabaena sp. to salt stress

Summary The response of a salt-tolerantWestiellopsis prolifica ARM 366 and a sensitiveAnabaena C-10 to NaCl was studied. While the former could tolerate up to 400mm NaCl, the latter was highly sensitive to concentrations above 50mm NaCl. Under salt stress, the tolerantW. prolifica showed an increased nitrogen demand as exemplified by high nitrogenase activity, and growth inhibition at higher concentrations of NaCl did not appear to be a direct consequence of inhibition of nitrogen fixation. One of the striking responses to Na⁺ challenge by the tolerantW. prolifica was the excess production of extracellular polysaccharides which adsorbed the bulk of the Na⁺. The influx of Na⁺ into the cell was comparatively small.

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Resumen Se estudió la respuesta al NaCl deWestiellopsis prolifica ARM 366, organismo tolerante a la sal, y deAnabaena C-10 que es sensible. El primero pudo tolerar concentraciones de hasta 400mm NaCl, sin embargo Anabaena C-10 se mostró extremadament sensible a concentraciones superiores a 50mm de NaCl. Bajo condiciones de estrés salinoW. prolifica mostró un incremento en la demanda de N₂, como se deduce de la elevada actividad nitrogenásica; la inhibición del crecimiento a concentraciones superiores de NaCl no se pudo relacionar directamente con la inhibición de la fijación de nitrógeno. Una de las respuestas más sorprendentes deW. prolifica frente al Na⁺ fue la producción de un exceso de polisaccaridos extracelulares que absorbieron gran parte del Na⁺. El flujo de Na⁺ hacia la célula fue, pues, comparativemente pequeño.

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Résumé On a étudié la réponse au NaCl d'une souche halo-tolérante deWestiellopsis prolifica (ARM 366) et, d'une souche halo-sensible d'Anabaena (C-10). Alors que la première tolère jusqu'a 400mm NaCl, la seconde est hautement sensible aux concentrations supérieures à 50mm. Soumise à un choc de salinité, la souche halo-tolérante deW. prolifica présente une demande d'azote accrue, comme le montre une activité nitrogénase élevée, et l'inhibition de la croissance par une forte concentration en NaCl ne parait pas être la conséquence directe d'une inhibition de la fixation d'azote. Une réponse remarquable deW. prolifica à l'agression par Na⁺ est la production en excès de polysaccharides exo-cellulaires, qui absorbent la plus grande partie du Na⁺. La pénétration de Na⁺ dans la cellule est relativement faible.

     

Estudio micologico de un caso de Micetoma por Cephalosporium en Puerto Rico

Resumen Hemos descrito un Cephalosporium como agente etiológico en un caso de Pié de Madura puertorriqueño. Este hongo crece bien en los medios ordinarios de laboratorio produciendo colonias características y dando origen a un pigmento difusible cuyo color varía de acuerdo con los ingredientes dal agar. Al examen microscópico, los cultivos nos revelan un gran número de conidios alargados que se caracterizan por su gran tamaño, su pronunciado eneorvamiento y su tabicación interna, a más de lo cual se notan muchos clamidosporos de morfología variadísima y, finalmente, estructuras nodulares muy semejantes a las que se desarrollan en ciertas especies del género Trichophyton. La identificatión específica del parásito ha quedado pendiente hasta tanto pueda hacerse un estudio comparativo de otras tres especies cefalospóricas que parecen guardar estrecha analogía con la nuestra a juzgar por las descripciones que de aquellas se han hecho.Sea cual fuere su posición sistemática, la especie aquí descrita no había sido encontrada nunca antes en las lesiones del micetoma. La literatura médica solamente registra otro caso de esa enfermedad producido por Cephalosporium. Se trataba entonces de una infectión contraida en el Brasil que tuvo como agente etiológico en parásito, distinto del nuestro, el cual fué estudiado y clasificado por Leão y Lobo como especie nueva : el Cephalosporium recifei.     

Fed-batch two-phase production of alanine by a metabolically engineered Escherichia coli

dl-Alanine was produced from glucose in an Escherichia coli pfl pps poxB ldhA aceEF pTrc99A-alaD strain which lacked pyruvate-formate lyase, phosphoenolpyruvate (PEP) synthase, pyruvate oxidase, lactate dehydogenase, components of the pyruvate dehydogenase complex and over-produced alanine dehydrogenase (ALD). A two-phase process was developed with cell growth under aerobic conditions followed by alanine production under anaerobic conditions. Using the batch mode, cells grew to 5.3 g/l in 9 h with the accumulation of 6–10 g acetate/l, and under subsequent anaerobic conditions achieved 34 g alanine/l in 13 h with a yield of 0.86 g/g glucose. Using the fed-batch mode at μ = 0.15 h⁻¹, only about 1 g acetate/l formed in the 25 h required for the cells to reach 5.6 g/l, and 88 g alanine/l accumulated during the subsequent 23 h. This fed-batch process attained an alanine volumetric productivity of 4 g/lh during the production phase, and a yield that was essentially 1 g/g.     

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.

     

Des considérations écologiques peuvent-elles apporter une contribution a la connaissance du cycle biologique des colonies deCremastogaster (Hymenoptera-Formicoidea)?

Resumen Ciertos nùmeros de luchos experimentales y de observación de cuatro especies deCremastogaster: Cr. auberti, Cr. scutellaris (de Francia);Cr. vandeli, Cr. skounensis (de Cambodia) han conducido a formular una hipotesis acerca del ciclo biologico de ciertos especies deCremastogaster.Hay alternancia entre dos generaciones, una agama reproduciéndose por partenogenesis, otra sexuada. La generación sexuada comprende machos y hembras aladas destinadas al origen para la diseminación del especie aprovechando las condiciones ecológicas favorables (vuelo de enjambre). La descendencia normal de las hembras est a constituida por obreras (sin embargo puede haber en las hembras partenogenesis arrhenotoque accidental del tipo himenoptere). La generación agama comprende solo hembras apteras (obreras) y està destinada para la conservación del especie durante las estationes con condiciones ecológicas defavorables. Esas obreras son poco prolificas y ciertamente solo por un breve periódo de su vida. La generación asexuada vuelve a dar una generación sexuada por partenogenesis deuterotoque (La deuterotoquia està bajo la influencia de factores troficos y, por consiguiente, ecológicos).

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Summary Some number of expérimental samples and observations on for species ofCremastogaster: Cr. auberti; Cr. scutellaris (of France);Cr. vandeli, Cr. skounensis (of Cambodia) brings to formule hypothesis on biological cycle of some species ofCremastogaster.There is alternance between two generations, the one without sexual types, which reproduces by parthenogenesis, the other with sexual types. The sexual generation comports winged males and winged females for dispersion of species (in beginning) which profit of ecological favourable conditions (wing of swarming). The normal descendance of females is constitued by workers (however by males it can existe accidental arrhenotocal parthenogenesis of Hymenopteral type). The without sexuals generation comports only wingless females (workers) and it is destined to conserve the species during seasons under unfavourable ecological conditions. These workers are not to much prolific and certainly during a short time of their life. The asexual generation gives again sexual generation by deuterotocal parthenogenesis (Deuterotoquia is under the influence of trophic and, from this fact, ecological factors).

     

Cinco cepas de Prototheca de origen humano

Resumen Se han aislado cinco cepas dePrototheca de muestras de origen humano: raspado vulvar, tinea corporis, secreción de oído, plieques de la mano, onicopatía inespecífica. Se estudian los caracteres morfológicos (propiedades asimilativas, termofilia) de estas cepas. Dos de ellas pertenecen a la especiePrototheca zopfii. Dos a la especieP. wickerhamii y la quinta es unaP. trispora. El autor hace resalter la frecuencia cada vez mayor con que se viene aislando este género de algas incoloras en el material patológico y la necesidad de su reconocimiento ya que son fácilmente confundibles con las levaduras.

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FivePrototheca strains of human origin are isolated. Morphological and physiological characters are studied. Two strains belong to the speciesPrototheca zophii, two to the speciesP. wickerhamii and the other one toProtheca trispora. The author emphasizes the frequency, with which this genus of chloric algae are lately reported, and the importance of their identification easily confused with that of the yeasts.Ultimamente hemos venido encontrando cada vez con mayor frecuencia en el material clínico, ejemplares del género Prototheca. La ubicación taxonómica de este género es aún dudosa; se le ha considerado un grupo intermedio entre algas incoloras y hongos (Tubaki, 1959). Nuestras 5 cepas han sido encontradas en un lapso de cuatro años, de un total de 320 muestras.

     

Pichia faecalis nov. comb.

In this paper, the auther re-examinesKloeckera faecalis Batista &Silveria 1959 and transfers to genusPichia.

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Resumen El autor hace una revisión de la levaduraKloeckera faecalis llevandola al géneroPichia.

     

Pichia faecalis nov. comb. revisión de una cepa de levadura nombradaKloeckera faecalis Batista et Silveria, 1959

In this paper, the author re-examinesKloeckera faecalis Bat. &Sil. 1959 and transfers to genusPichia.

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Resume El autor hace una revisión de la levaduraKloeckera faecalis llevàndola al géneroPichia.