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J Mora

Publications and source records attributed to J Mora.

At least 145 records · Page 8Linked to original sources
PubMed1983

Nitrogen source regulates glutamate dehydrogenase NADP synthesis in Neurospora crassa.

Neurospora crassa glutamate dehydrogenase-NADP (EC 1.3.1.3) has a higher activity when mycelium is grown on ammonium or nitrate as nitrogen source than when grown on glutamate or glutamine. Quantitative immunoelectrophoresis established that, under all conditions, enzyme activity corresponded to enzyme concentration. Isotope incorporation studies demonstrated that the nitrogen source exerts its regulation at the level of de novo enzyme synthesis.

Glutamate Dehydrogenase↗
PubMed1983

Neurospora crassa mutant impaired in glutamine regulation.

The final products of the catabolism of arginine that can be utilized as nitrogen sources by Neurospora crassa are ammonium, glutamic acid, and glutamine. Of these compounds, only glutamine represses arginase and glutamine synthetase. We report here the isolation and characterization of a mutant of N. crassa whose arginase, glutamine synthetase, and amino acid accumulations are resistant to glutamine repression (glnI). This mutant has a greater capacity than the wild type (glns) to accumulate most of the arginine and some of the glutamine in osmotically sensitive compartments while growing exponentially. Nonetheless, the major part of the glutamine remains soluble and metabolically available for repression. We propose that the lower repression of glutamine synthetase by glutamine in this mutant could be a necessary condition for sustaining the higher flow of nitrogen for the accumulation of amino acids observed in ammonium excess and that, if glutamine is the nitrogen signal that regulates the arginine accumulation of the vesicle, the glnr mutant has also escaped this control. Finally, in the glnr mutant, some glutamine resynthesis is necessary for arginine biosynthesis and accumulation.

Amino Acids↗
PubMed1983

Genetic and biochemical characterization of glutamine synthetase from Neurospora crassa glutamine auxotrophs and their revertants.

In this paper we present the isolation and characterization of glutamine auxotrophs of Neurospora crassa and their revertants. The results show that although various enrichment procedures were used, we found only two types of auxotrophs. Genetic crosses performed between the different mutants showed that the mutations responsible for their phenotypes were highly linked and probably affected the same gene. The biochemical characterization of the glutamine synthetase polypeptides of the different mutants showed that both types contained the alpha monomer. However, in place of the normal beta monomer, each type had a new polypeptide differing from normal beta either in its molecular weight or in its isoelectric point. On the other hand, the revertants had only the alpha monomer and were capable of growing without glutamine. On the basis of these data, we propose that the lack of glutamine synthetase activity in the auxotrophs is due to the interaction of the altered beta with the alpha monomer, and as a consequence the alpha monomer of the revertants regains its activity because of the absence of the altered beta.

Centrifugation, Density Gradient↗
PubMed1983

Local blood flow, oxygen tension, and oxygen consumption in the rat spinal cord. Part 1: Oxygen metabolism and neuronal function.

A reliable and reproducible microelectrode technique provided consistent simultaneous measurements of local spinal cord blood flow (local SCBF), tissue oxygen tension (TO2), and tissue oxygen consumption (TO2C) in the rat. Local SCBF was measured by the hydrogen clearance technique, local TO2 was measured polarographically, and local TO2C was calculated from the declining slope of local TO2 following temporary arrest of local SCBF. Mean local TO2 values varied only slightly between gray and white matter, while local TO2C and SCBF maintained a 3 to 1 ratio between gray and white matter areas. Measurements were also made of these parameters in specific white matter tracts and laminae of Rexed. Local white matter SCBF was relatively homogeneous throughout the ventral, lateral, and dorsal funiculi. Gray matter blood flow was more variable with topography. The highest local SCBF was recorded in areas rich in internuncial neurons. The somatic motor regions had values slightly higher than recorded in sensory regions.

Animals↗
PubMed1983

Local blood flow, oxygen tension, and oxygen consumption in the rat spinal cord. Part 2: Relation to segmental level.

Using a reliable and reproducible microelectrode technique, consistent simultaneous measurements of local spinal cord blood flow (SCBF), tissue oxygen tension, and tissue oxygen consumption were made at cervical, thoracic, and lumbar levels in the rat spinal cord. These observations showed that the metabolic state is maintained constant along the cord, despite significant variations in vasculature. The physiological and anatomical aspects of these findings are discussed.

Animals↗
PubMed1982

Surveillance study of pediatric patients receiving respiratory therapy, using the Bain circuit system.

The Bain circuit, which has been used previously for anesthesia, was adopted for long-term mechanical ventilation for infants and children in a pediatric intensive care unit. When this ventilation system is used, humidifier temperatures must be kept at temperatures significantly lower than the recommended 50 degrees C to avoid excessive airway temperatures and thermal injury. Routine infection control measures such as daily changing of respiratory therapy equipment and the use of sterile water in Cascade humidifiers were enforced. It was demonstrated that the use of the Bain circuit with a Cascade humidifier kept at a temperature of 36.9 degrees +/- 1.9 degrees C was not associated with nosocomial infection of the lower respiratory tract.

Acinetobacter↗
PubMed1982

Physiology of ammonium assimilation in Neurospora crassa.

In Neurospora crassa the assimilation of high and low concentrations of ammonium occurs by two different pathways. When the fungi are growing exponentially on ammonium excess, this compound is fixed by a glutamic dehydrogenase and an octameric glutamine synthetase (GS). The synthesis of this GS polypeptide (beta) is regulated by the nitrogen source present in excess; being higher on glutamate, intermediate on ammonium, and lower on glutamine. When N. crassa is growing in fed-batch ammonium-limited cultures a different polypeptide of GS (alpha), arranged as a tetramer, is synthesized. In both conditions synthesis in vivo correlates with the data obtained with an in vitro translation system primed with N. crassa RNA. This different expression of alpha and beta GS polypeptides was also observed when the cultures were shifted from excess to low nitrogen, and vice versa. By agarose gel electrophoresis in the presence of methylmercury hydroxide, some separation of different mRNAs that direct the in vitro synthesis of alpha and beta GS polypeptides has been accomplished. Data are presented that establish the operation of the tetrameric alpha GS and of glutamate synthase in the assimilation of ammonium in low concentration.

Ammonium Chloride↗
PubMed1981

Modified Mapleson D system for long-term mechanical ventilation of infants and children.

The Bain circuit, a modified Mapleson D system, is a lightweight, simple circuit which has been used primarily for anesthesia. This report describes its use for long-term mechanical ventilation for infants and children. The use of this circuit improved warming of inspired gas reducing patient heat loss and, additionally, it was believed to have resulted in increased humidity of the inspired gas. There were no instances of accidental extubation nor were any of the endotracheal tubes blocked by inspissated secretions. Temperatures of the humidifiers had to be lower than conventionally recommended but this did not result in any nosocomial infections. This circuit is an effective and safe circuit to use for long-term mechanical ventilation of children.

Child↗
PubMed1979

Glutamine metabolism in nitrogen-starved conidia of Neurospora crassa.

During nitrogen deprivation, de novo synthesis of glutamine synthetase was induced in non-growing conidia of Neurospora crassa. When ammonia or glutamine was added to conidia which had been deprived of nitrogen, glutamine and arginine accumulated at a higher rate than in condia not deprived of nitrogen. The degradation of exogenous glutamine to glutamate is apparently a necessary step in the accumulation of glutamine and arginine within the conidia. In non-growing conidia, a cycle probably operates in which glutamine is degraded and resynthesized. The advantages of such a cycle would be that the carbon and nitrogen could be used to synthesize amino acids in general, as well as for the synthesis and accumulation of arginine and/or glutamine in particular.

Arginine↗
PubMed1979

Immunochemical characterization of glutamine synthetase from Neurospora crassa glutamine auxotrophs.

Glutamine synthetase derived from two Neurospora crassa glutamine auxotrophs was characterized. Previous genetic studies indicated that the mutations responsible for the glutamine auxotrophy are allelic and map in chromosome V. When measured in crude extracts, both mutant strains had lower glutamine synthetase specific activity than that found in the wild-type strain. The enzyme from both auxotrophs and the wild-type strain was partially purified from cultures grown on glutamine as the sole nitrogen source, and immunochemical studies were performed in crude extracts and purified fractions. Quantitative rocket immunoelectrophoresis indicated that the activity per enzyme molecule is lower in the mutants than in the wild-type strain; immunoelectrophoresis and immunochemical titration of enzyme activity demonstrated structural differences between the enzymes from both auxotrophs. On the other hand, the monomer of glutamine synthetase of both mutants was found to be of a molecular weight similar to that of the wild-type strain. These data indicate that the mutations are located in the structural gene of N. crassa glutamine synthetase.

Genes↗
PubMed1978

Nitrogen regulation of glutamine synthetase in Neurospora crassa.

A higher activity of glutamine synthetase (EC 6.3.1.2) was found in Neurospora crassa when NH4+ was limiting as nitrogen source than when glutamate was limiting. When glutamate, glutamine or NH4+ were in excess, a lower activity was found. Immunological titration and sucrose gradient sedimentation of the enzyme established that under all these conditions enzyme activity corresponded to enzyme concentration and that the octamer was the predominant oligomeric form. When N. crassa was shifted from nitrogen-limiting substrates to excess product as nitrogen source, the concentration of glutamine synthetase was adjusted with kinetics that closely followed dilution by growth. When grown on limiting amounts of glutamate, a lower oligomer was present in addition to the octameric form of the enzyme. When the culture was shifted to excess NH4+, glutamine accululated at a high rate; nevertheless, there was only a slow decrease in enzyme activity and no modification of the oligomeric pattern.

Culture Media↗
PubMed1978

Genetics and physiology of Neurospora crassa glutamine auxotrophs.

This work reports on the isolation and characterization of two glutamine auxotrophs in Neurospora crassa. The mutations responsible for the glutamine-requiring phenotype were very closely linked, and one of them proved to be recessive to wild type. The mutations impaired the conversion of glutamic acid to glutamine and resulted in changes of both the activity and oligomeric structure of the enzyme glutamine synthetase.

Ammonia↗
PubMed1978

Nitrogen source regulates glutamine synthetase mRNA levels in Neurospora crassa.

Neurospora crassa glutamine synthetase mRNA was measured by its capacity to direct the synthesis of the specific protein in a cell-free system derived from rabbit reticulocytes. N. crassa cultures grown on glutamate as the sole nitrogen source had higher mRNA activities than did those grown on glutamine. The differences were about 10-fold when polysomal RNA was used for translation and about 5-fold when either total cellular RNA or polyadenylic acid-enriched cellular RNA was used. These data indicate that in exponentially growing N. crassa, the nitrogen source regulates glutamine synthetase by adjusting specific mRNA levels.

Cell-Free System↗