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Biomedical subjects

H Bertrand

Publications and source records attributed to H Bertrand.

At least 55 records · Page 3Linked to original sources

Unstable mitochondrial DNA in natural-death nuclear mutants of Neurospora crassa.

The natural-death mutant of Neurospora crassa has an accelerated senescence phenotype caused by a recessive mutation, nd, in a nuclear gene that is located in linkage group I. An examination of mitochondrial functions, however, revealed that the mutant has phenotypic and molecular defects similar to those commonly associated with maternally transmitted fungal senescence syndromes, including (i) deficiencies in cytochromes aa3 and b; (ii) a deficit in small subunits of mitochondrial ribosomes, and hence defective mitochondrial protein synthesis; and (iii) accumulation of gross rearrangements, including large deletions, in the mitochondrial chromosome of vegetatively propagated cells. These traits indicate that the nd+ allele codes for a function that is essential for stable maintenance of the mitochondrial chromosome, possibly a protein involved in replication, repair, or recombination.

Cell Survival↗

Cytochrome oxidase subunit V gene of Neurospora crassa: DNA sequences, chromosomal mapping, and evidence that the cya-4 locus specifies the structural gene for subunit V.

The sequences of cDNA and genomic DNA clones for Neurospora cytochrome oxidase subunit V show that the protein is synthesized as a 171-amino-acid precursor containing a 27-amino-acid N-terminal extension. The subunit V protein sequence is 34% identical to that of Saccharomyces cerevisiae subunit V; these proteins, as well as the corresponding bovine subunit, subunit IV, contain a single hydrophobic domain which most likely spans the inner mitochondrial membrane. The Neurospora crassa subunit V gene (cox5) contains two introns, 398 and 68 nucleotides long, which share the conserved intron boundaries 5'GTRNGT...CAG3' and the internal consensus sequence ACTRACA. Two short sequences, YGCCAG and YCCGTTY, are repeated four times each in the cox5 gene upstream of the mRNA 5' termini. The cox5 mRNA 5' ends are heterogeneous, with the major mRNA 5' end located 144 to 147 nucleotides upstream from the translational start site. The mRNA contains a 3'-untranslated region of 186 to 187 nucleotides. Using restriction-fragment-length polymorphism, we mapped the cox5 gene to linkage group IIR, close to the arg-5 locus. Since one of the mutations causing cytochrome oxidase deficiency in N. crassa, cya-4-23, also maps there, we transformed the cya-4-23 strain with the wild-type cox5 gene. In contrast to cya-4-23 cells, which grow slowly, cox5 transformants grew quickly, contained cytochrome oxidase, and had 8- to 11-fold-higher levels of subunit V in their mitochondria. These data suggest (i) that the cya-4 locus in N. crassa specifies structural information for cytochrome oxidase subunit V and (ii) that, in N. crassa, as in S. cerevisiae, deficiencies in the production of nuclearly encoded cytochrome oxidase subunits result in deficiency in cytochrome oxidase activity. Finally, we show that the lower levels of subunit V in cya-4-23 cells are most likely due to substantially reduced levels of translatable subunit V mRNA.

Amino Acid Sequence↗

Immunological identification of the alternative oxidase of Neurospora crassa mitochondria.

Neurospora crassa mitochondria use a branched electron transport system in which one branch is a conventional cytochrome system and the other is an alternative cyanide-resistant, hydroxamic acid-sensitive oxidase that is induced when the cytochrome system is impaired. We used a monoclonal antibody to the alternative oxidase of the higher plant Sauromatum guttatum to identify a similar set of related polypeptides (Mr, 36,500 and 37,000) that was associated with the alternative oxidase activity of N. crassa mitochondria. These polypeptides were not present constitutively in the mitochondria of a wild-type N. crassa strain, but were produced in high amounts under conditions that induced alternative oxidase activity. Under the same conditions, mutants in the aod-1 gene, with one exception, produced apparently inactive alternative oxidase polypeptides, whereas mutants in the aod-2 gene failed to produce these polypeptides. The latter findings support the hypothesis that aod-1 is a structural gene for the alternative oxidase and that the aod-2 gene encodes a component that is required for induction of alternative oxidase activity. Finally, our results indicate that the alternative oxidase is highly conserved, even between plant and fungal species.

Antibodies, Monoclonal↗

[Infectious complications after surgery of esophageal pathology. Apropos of 60 cases].

Infections after oesophageal surgery are studied on sixty patients who received perioperatively two antibiotics (cefoxitin and amikacin). Postoperative infection rate is 62% (pneumopathies: 27%, leakage of cervical anastomoses: 17%). Different parameters which can induce postoperative infection are analyzed. The only significative data are the duration of total parenteral nutrition and of intensive care stay. The commonest isolated organisms are gram negative bacilli (61%) and streptococci (30%). Yeasts infections are frequent, and significantly correlated with antibiotic treatment duration.

Aged↗

[The value of monitoring arterial oxygenation saturation by pulse oximetry in neuroleptanalgesia].

Anaesthesia can induce hypoxaemia. Pulse oximetry gives continuous non invasive monitoring of arterial oxygen saturation. No arterial puncture is needed. Ninety-four patients were monitored by pulse oximetry during neuroleptanalgesia for colonoscopy. Eighteen patients showed desaturation episodes of less than 90%, eight linked to opioid-induced respiratory depression. In all patients, pulse oximetry gave clinicians an immediate awareness of the incident. Treatment was facilitated. Pulse oximetry made anaesthesia safer.

Blood Gas Monitoring, Transcutaneous↗

An extrachromosomal plasmid is the etiological precursor of kalDNA insertion sequences in the mitochondrial chromosome of senescent neurospora.

In the kalilo strains of N. intermedia, senescence is initiated by insertion of a 9.0 kb foreign nucleotide sequence, kalDNA, into mitochondrial DNA. A 9.0 kb linear DNA plasmid that is structurally homologous to the mitochondrial kalDNA insertion sequences exists in high copy numbers in close association with the nuclei of presenescent and senescent kalilo cells, but is not present in cells of long-lived normal strains. The free kalilo plasmid has not been detected in mitochondria, suggesting that the element does not contain a mitochondrial origin of replication. Unexpectedly, the nuclear plasmid, like the mitochondrial insertion element, follows a strict pattern of maternal inheritance. We surmise that the extramitochondrial plasmid is the etiological precursor of the kalDNA insertion sequences that appear in the mtDNAs of senescent cell lines and conclude that the kalilo element induces senescence because it is a mutator of mitochondrial genes.

DNA Replication↗

Inversions and recombinations in mitochondrial DNA of the (SG-1) cytoplasmic mutant in two Neurospora species.

The mitochondrial DNAs of [SG-1] cytoplasmically-mutant and wild-type strains of Neurospora crassa and Neurospora sitophila were examined by comparative restriction endonuclease analyses. The mtDNA of N. sitophila wild type of Whitehouse differs from type II mtDNA of N. crassa by insertions of 3.3 kb in EcoRI-9, and 1.2 kb in EcoRI-3, and a deletion of 1.1 kb in EcoRI-5. These DNA heteromorphisms provided convenient markers for tracing N. crassa [SG-1] mtDNA during and after its transfer into N. sitophila. The [SG-1] cytoplasmic mutant in both N. crassa and N. sitophila has a distinctive inversion that connects the fragment EcoRI-4 with HindIII-10a. The [SG-1] mtDNA from N. crassa remained essentially intact after it was transferred by crosses into N. sitophila. In each species, a unique second inversion occurred in the [SG-1] mtDNA after the transfer was made. In N. sitophila, polar recombination in heteroplasmons between [SG-1] and wild-type preferentially yields strains with mtDNAs that contain the maximum possible number of insertions in the cob and co-1 loci of the EcoRI-3 region of the mitochondrial chromosome.

Chromosome Inversion↗

Insertion of a foreign nucleotide sequence into mitochondrial DNA causes senescence in Neurospora intermedia.

The kalilo variants of Neurospora contain a cytoplasmic genetic factor that causes senescence. This factor is a 9.0 kb transposable element (kalDNA) that lacks nucleotide sequence homology with mtDNA and is inserted into the mitochondrial chromosome, often at sites located within the open reading frame in the intron-DNA of the mitochondrial 25S-rRNA gene. Genomes containing the "foreign" DNA insert accumulate during growth, and death occurs as the cells become deficient in functional large and small subunits of mitochondrial ribosomes. The kalDNA transposon may be an "activator" element that causes breaks in mtDNA. Nonsenescing [+] strains of Neurospora do not contain kalDNA.

Base Sequence↗

In vivo neutralization of low-molecular weight heparin fraction CY 216 by protamine.

The neutralization in vivo of a low molecular weight heparin by protamine was investigated. Large doses and excessive doses of intravenously administered CY 216 were studied. An intravenous injection of protamine given 10 minutes after the administration of CY 216 did not cause the studied biologic parameters to return to normal levels, but merely attenuated them, whatever the protamine dosage tested. In contrast, the bleeding time and the volume of blood loss resumed normal values that were close to those observed in the controls. This dissociation of actions cannot be explained at present. The ratio of protamine to CY 216 dosage that produced the best results was 1 antiheparin U of protamine to 2 anti-Xa U of CY 216. Nevertheless CY 216 appeared to have a small hemorrhagic potential.

Animals↗

(2'-5') Oligoadenylate synthetase in the maturation of rabbit reticulocytes.

The (2'-5') oligoadenylate synthetase normally found in interferon-treated cells has also been detected in considerable amounts in normal rabbit reticulocytes not exposed to interferon. The activity of this enzyme has been followed during the development of the reticulocytes to erythrocytes. A high level was found just after the formation of reticulocytes and this activity decayed with a half-life of about 3 days. In lymphocytes the (2'-5') oligoadenylate synthetase was found to stay at a constant level, which indicates the absence of interferon in the plasma.

2',5'-Oligoadenylate Synthetase↗

Age changes in hepatic metabolic characteristics and their modulation by dietary manipulation.

Starting at 6 weeks of age, male Fischer 344 rats were provided five different dietary regimens: group 1, fed ad libitum; group 2, restricted to 60% of the food intake of group 1; group 3, restricted to 60% of the food intake of group 1 until 6 months of age and then fed ad libitum; group 4, fed ad libitum until 6 months of age and then restricted to 60% of the food intake of group 1; group 5, given the same caloric intake as group 1 but 60% of the protein intake. The weight of the liver was maintained at about 2.5% of body weight over the wide range of body weight, age (6 through 30 months of age) and diets of this study. Liver cholesterol concentration increased with age in the rats of group 1 but not in the other groups; the hepatic cholesterol concentration was lower in the rats of groups 2 and 4 than in the others. The liver microsomal 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase activity was higher in 4-month-old rats than in rats 12 months of age and older; also, the microsomal activity from group 2 and group 4 rats was higher than that from rats of the other groups. Liver triglyceride levels increased with age in groups 1, 2, 3 and 5 but not in group 4. At all ages, postabsorptive (15 h without food) hepatic glycogen concentrations were much higher in groups 2 and 4 than in the other groups. Liver phospholipid concentration was not affected by age or diet. Basal adenylate cyclase activity in liver homogenates increased with age in all but group 4, but hormone-stimulated adenylate cyclase activity was little affected by age or diet. The age-related hepatic changes in lipids and in hormone responsiveness noted in the present study were much less marked than those previously found in blood and adipose tissue.

Adenylyl Cyclases↗

RNA splicing in Neurospora mitochondria: nuclear mutants defective in both splicing and 3' end synthesis of the large rRNA.

We have identified nuclear mutants of Neurospora that are defective in splicing the mitochondrial large rRNA and that accumulate unspliced pre-rRNA (35S RNA). In cyt-4 mutants, the unspliced pre-rRNA contains short 3' end extensions (110 nucleotides) that are not present in pre-rRNAs from the other mutants. This and other characteristics suggest that the cyt-4 mutants may be primarily defective in 3' end synthesis and the RNA splicing defect occurs secondarily as a result of impaired RNA folding. The cyt-4 mutants also accumulate a "short" intron RNA and small exon RNAs that may reflect aberrant RNA cleavages. The 5' end of the short intron is about 285 nucleotides downstream from the 5' splice site at or near the base of the "central hairpin", a putative intermediate in folding of the pre-rRNA. Furthermore, the aberrant cleavage sites are immediately after a six nucleotide sequence (GAUAAU) homologous to the final splice junction (GAU/AAC).

Base Sequence↗

Temporal and compositional dietary restrictions modulate age-related changes in serum lipids.

Life-lon restriction of food, which markedly increases the length of life of rats, also delays the occurrence of age-related physiological deteriorations including the age-related changes in serum lipids. The aim of this study was to explore the effects of 1) food restriction limited to early life, 2) food restriction initiated in adult life and 3) restriction of protein without restriction of calories on the age-related changes in serum lipids. Food restriction started at 6 months of age was as effective as food restriction started at 6 weeks of age in modulating the age-related changes in serum lipids, whereas food restriction from 6 weeks to 6 months of age and restriction of protein but not calories were much less effective. Changes in the rate of very low density lipoprotein-triglyceride removal from the blood were not responsible for the age-related increase in triglyceride concentration or for the modulation of this increase by the dietary regimens. Age-related changes in serum or plasma levels of glucose, ketone bodies, insulin and glucagon were also measured and the modulating effects of the dietary regimens studied; possible relationships between the concentrations of these substances and the concentrations of the serum lipids are discussed.

Aging↗

Studies on a highly active anticoagulant fraction of high molecular weight isolated from porcine sodium heparin.

We have studied heparin fractionation using gel filtration and ion-exchange chromatographic methods. The starting material was commercial grade porcine mucosal sodium heparin (PSH). The fractionation was monitored employing synthetic substrates for assaying both antithrombin (with H-D-Phe-Pip-Arg-pNA ; S-2238) and anti-FXa (with Bz-Ileu-Glu-Gly-Arg-pNA ; S-2222) activities. The resulting fractions were evaluated in different amidolytic and coagulation methods used to determine heparin potency by comparison with PSH. By gel filtration of PSH on Ultrogel Aca 54, both strong anti-FXa and antithrombin activities were associated with the fractions eluted in the high molecular weight range (MW congruent to 20 x 10(3)). These fractions also had potent anticoagulant action when assayed by conventional clotting methods. PSH was also subjected to fractionation by an ion-exchange technique (DEAE-Sephacel) with increasing salt molarity. The patterns for antithrombin and anti-FXa activities were again closely related, if not identical. Four fractions were usually distinguished, with respectively negligible, intermediate, high and very high activities when compared to PSH. The very highly active fraction (HAF), approximately 15% by weight, was eluted at high salt molarity (greater than 0.8 M NaCl). On a weight basis its anticoagulant activity was congruent to 2-3 times that of PSH as determined by amidolytic as well as clotting methods. Intravenous injection of HAF to rabbits and dogs (1.0 and 2.5 mg/kg) produced a much stronger anticoagulant response than PSH, also showing an effect which persisted for a longer duration.

Animals↗