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R Durand

Publications and source records attributed to R Durand.

At least 19 recordsLinked to original sources
PubMed1992

Sequence of the phosphoenolpyruvate carboxykinase-encoding cDNA from the rumen anaerobic fungus Neocallimastix frontalis: comparison of the amino acid sequence with animals and yeast.

The nucleotide sequence of the cDNA of the phosphoenolpyruvate carboxykinase-encoding gene from the fungus Neocallimastix frontalis, was determined. The deduced amino acid sequence (608 residues) and the predicted protein structure were compared to their counterparts in animals and yeast. Catalytic regions (substrate-binding site and nucleotide-binding domains) are highly conserved among fungal and animal organisms. The yeast sequence showed no similarity to the fungal sequence.

Amino Acid Sequence↗
PubMed1991

Molecular cloning of genes from the rumen anaerobic fungus Neocallimastix frontalis: expression during hydrolase induction.

Glycoside and polysaccharide hydrolase production by the rumen anaerobic fungus, Neocalimastix frontalis is induced by the presence of crystalline cellulose. A differential screening of a cDNA library was used to isolate DNA sequences transcribed at high levels under growth conditions which induce enzyme production. Seven clones were isolated that preferentially hybridized to the induced cDNA probe versus the non-induced cDNA probe. Southern analysis showed that a cDNA clone (118) hybridized to a DNA probe encoding part of the exo-cellobiohydrolase I (CBH I) gene of Trichoderma reesei. Northern analysis demonstrated that the cDNA 118 was transcribed to yield a 2.1 kb RNA. This transcript was induced in the presence of cellulose.

Anaerobiosis↗
PubMed1991

The effect of hydralazine on blood flow and misonidazole toxicity in human tumour xenografts.

The effect of post-irradiation hypoxia induced by 5 or 30 mg/kg hydralazine has been studied in three human tumour xenografts (two rectocolic adenocarcinomas and one melanoma) treated with two doses of misonidazole similar to those used in patients (0.1 and 0.2 mg/g). Only a small sensitization was detected using an in vitro colony assay. These results are in marked contrast to the results obtained with rodent tumours. This difference between human tumour xenografts and rodent tumours might be explained by differences in the reduction of tumour blood flow after hydralazine administration (5 and/or 10 mg/kg). Using the laser Doppler technique, the tumour blood flow reduction was 33% and 25% of the control for NA11 and HRT18 tumours, respectively. In contrast, hydralazine induced a 60-70% reduction in blood flow in the murine SCCVII tumour. Using the fluorescent marker Hoechst 33342, the reduction in perfusion was again more pronounced in the murine tumour as compared to the Na11 and HRT18 xenografts. The differences between human tumour xenografts and rodent tumours are not linked to the mouse strain used (nude versus C3H) nor to a tumour bed effect.

Animals↗
PubMed1990

Evaluating the effect of a human values seminar series on ethical attitudes toward resuscitation among pediatric residents.

A human values seminar series was designed to provide training in medical ethics for residents in pediatrics. Attitudes of residents toward resuscitation of children were surveyed before and after the seminar series. Nonpediatric residents rotating through pediatrics served as control subjects. Although both groups of residents demonstrated a significant change in attitude on six scales at the end of 1 year of training, pediatricians and nonpediatricians differed from each other with respect to only one of these scales. After the seminar series, pediatric residents expressed decreased willingness to resuscitate certain critically ill infants in the emergency room, whereas nonpediatricians expressed more willingness (P less than 0.009). Attitude change in decision making among pediatricians was significantly correlated with the consideration given to societal needs, sanctity of life, and anticipated poor morbidity (P less than 0.05). Among nonpediatricians, attitude change was significantly correlated with poor prognosis for survival (P less than 0.05), morbidity (P less than 0.05), or anticipated poor intellectual outcome (P less than 0.01). Attitude differences between the two groups of residents were unrelated to differences in preexisting attitudes or demographic factors. These findings show that attitudes about ethical decisions are influenced by both "maturation," or progression through a training program, and formal instruction. The finding that clinical attitudes change rapidly as residents progress through training confirms our previous findings and suggests that residency training programs should provide support and formal instruction in ethics during this time of change. The small change caused by this current method of instruction indicates that preliminary studies are needed to delineate the goals and the structure of future ethics courses.

Adult↗
PubMed1989

Gastrointestinal effects of water reuse for public park irrigation.

To investigate the gastrointestinal effects of employing recycled water as an irrigation source for urban public parks, we studied subjects active in parks irrigated with potable water, nonpotable water of wastewater origin, and nonpotable water of runoff origin. Wet grass conditions during activity and elevated densities of common indicator bacteria, but not exposure to nonpotable irrigation water per se, were found associated with an increased rate of gastrointestinal illness.

Adolescent↗
PubMed1986

RNA mapping on Drosophila mitochondrial DNA: precursors and template strands.

Drosophila melanogaster mitochondrial DNA (mtDNA) is closely related to the mammalian and amphibian mtDNA except for gene organization. In Drosophila, genes are distributed in clusters alternatively coded on each strand. Besides the eleven major foreseeable transcripts previously described (MERTEN and PARDUE, 1981, J. Mol. Biol., 153, 1-21), we have characterized two poly A+ transcripts, one major and one minor which could correspond respectively to the ND3 and ND6 reading frames, and 27 poly A+ minor transcripts (0.2 to greater than 3.2 kb) which are distributed along the mtDNA except in the rRNAs, ND 1 and A+ T rich regions. The mapping and length of 25 of these transcripts strongly suggest a precursor role. They would be processed at the level of tRNA or tRNA-like sequences. Most of them are transcribed from the template strand of each gene cluster and their distribution is in agreement with the hypothesis of several transcription origins and terminations located near the extremities of each gene cluster. Quantitatively our results show a large variation in each presumptive mature transcript compared to the other, even in a given gene cluster, suggesting a specific degradation of some of the mature transcripts.

Animals↗
PubMed1986

Longitudinal development in pediatric residents of attitudes toward neonatal resuscitation.

We used Guttman scaling procedures to devise a quantitative, reproducible measure among pediatric residents of attitude change concerning neonatal resuscitation. Preliminary cross-sectional testing of an incoming group of pediatric level 1 residents and graduating pediatric level 3 residents indicated that pediatric level 3 residents were more reluctant to resuscitate high-risk infants. This reluctance was not due to age differences. The pediatric level 1 residents were retested at the completion of each year of training. Residents showed significantly increased reluctance to resuscitate infants at the end of the first year of training and again at the end of the third year of training. These attitude changes were unrelated to gender, marital status, religious preference, or ethnic background. Data acquired both cross-sectionally and longitudinally indicated that attitudes toward neonatal resuscitation changed during residency training.

Attitude of Health Personnel↗
PubMed1986

Translation of polyuridylic acid in lysed mitochondria.

After osmotic shock with 50 mM Tricine buffer (pH 7.9), isolated mitochondria from D. Melanogaster embryos are treated with a low concentration of Triton X-100 (25 micrograms/mg of protein). The lysed mitochondria are still capable of RNA and protein synthesis. While incorporation of labeled precursor is often higher in lysed than in intact mitochondria, neosynthesized proteins exhibit similar electrophoretic patterns. Studies of labeled precursor incorporation in the presence of various effectors indicate a better accessibility to the translation machinery in lysed mitochondria than in intact mitochondria. Such a system has proven capable of translating an exogenous synthetic mRNA, i.e., poly (U).

Animals↗
PubMed1985

Mitochondrial DNA expression in Drosophila melanogaster: neosynthesized polypeptides in isolated mitochondria.

The expression of mitochondrial genome of D. melanogaster in isolated mitochondria was followed by incorporation of 35S methionine in neosynthesized polypeptides. A high level of protein synthesis was obtained after optimization of all the incubation parameters. Two kinds of energy-generating systems were used: an endogenous system where an oxidizable substrate were added for ATP synthesis; an exogenous system with an energy-rich compound for ATP regeneration, the latter proved to be the most effective. The effect of the oxidative phosphorylation uncoupler (Clccp), and an ATPase inhibitor (oligomycine) allow us to postulate the role of the electrochemical potential in the expression of the mitochondrial genome. Electrophoresis and autoradiography of neosynthesized mitochondrial proteins exhibits 18 to 24 protein bands, ranging from 6.5 to 65 Kd; incubation of KC 0% drosophila cells with 35S methionine and cycloheximide gave similar results. Both our results and those published elsewhere suggest that the expression of mitochondrial genome in higher organisms could be more complex than simple translation of the 13 genes presents on these genomes.

Animals↗
PubMed1984

Complex RNA chain elongation kinetics by wheat germ RNA polymerase II.

Kinetics of RNA chain elongation catalyzed by wheat germ RNA polymerase II have been studied using various synthetic DNA templates in the presence of excess dinucleotide monophosphate primers. With single- or double-stranded homopolymer templates, the double reciprocal plots 1/(velocity) as a function of 1/(nucleotide substrate) exhibit positive, negative or no curvature. With poly(dAT) as template, the mechanism of nucleoside monophosphate incorporation into RNA is not the ping-pong kinetic mechanism which was derived for E. coli RNA polymerase (6). Noncomplementary nucleoside triphosphates inhibit RNA transcription allosterically. Cordycepin triphosphate behaves as ATP, and not only inhibits AMP incorporation but also that of UMP and GMP on appropriate templates. The reason for this complex kinetic behavior is not yet understood. Possibilities are raised that there are several nucleoside triphosphate binding sites on wheat germ RNA polymerase II, that additional nucleoside triphosphate dependent enzymatic activities are required for reaction to occur or that the Km value for incorporation of a given nucleoside monophosphate into RNA is dependent on the length of the RNA chain and/or the nucleotide sequence surrounding the complementary base on the DNA template.

Kinetics↗
PubMed1984

[Isolation of the RNA-poly A+ fraction from mitochondria of Drosophila melanogaster embryos. Incorporation of methionine-35S into newly-synthesized proteins in whole and lysed mitochondria].

The poly A+ RNA fraction of embryos mitochondria of Drosophila melanogaster was prepared by fractionation by oligo dT-cellulose chromatography. It accounted for 5% of total insoluble RNA. 15 bands were identified by electrophoresis, from 500 to 2,000 base pairs. This poly A+ RNA fraction hybridizes with mitochondrial DNA clones. It contains mitochondrial transcripts: messenger RNA and the large ribosomal RNA. The sum of the transcripts (29 kb) exceed the coding capacities of the mitochondrial DNA. Some of the transcripts may be precursors, greater in size, of the mRNA corresponding to a gene. Our results are compared to those previously published (9, 11, 12). The relationship between mitochondrial transcripts and proteins was studied in whole and lysed isolated mitochondria. In the former, incubated in the presence of 35S-methionine, 20 to 30 radioactive bands were identified (M. Wt. 100 to 16 kd). This incorporation was totally inhibited by chloramphenicol. When the mitochondria were opened in the presence of detergent (lysis). 35S-methionine incorporation was markedly enhanced. In all cases the number of proteins identified was greater than the number of potential genes (12). This finding is discussed.

Animals↗
PubMed1984

Newborn ethical dilemmas: intensive care and intermediate care nursing attitudes.

A self-administered questionnaire concerning various neonatal ethical issues was completed by 39 ICU and 36 intermediate care unit (INT) nurses. Guttman scale analysis indicated that ICU nurses were more reluctant than INT nurses to resuscitate certain high-risk newborn infants. Work experience and a stated religious preference accentuated an INT nurse's disinclination to resuscitate high-risk babies. ICU nurses were more likely than INT nurses to favor passive and active euthanasia. Further, ICU nurses were more likely than INT nurses to view termination of life support for a sick infant as a necessity. ICU nurses were less likely than INT nurses to favor physician intervention in the care of all acutely sick neonates. ICU and INT nurses agreed, however, that the decision to terminate a baby's life support should be made jointly by the child's parents and physicians. Although differences in ICU and INT nursing attitudes may reflect quantitative and qualitative differences in work experience, these findings suggest a common need for emotional support and continuing education programs for nurses who care for high-risk newborns.

Adult↗
PubMed1983

Comparative transcription of right- and left-handed poly[d(G-C)] by wheat germ RNA polymerase II.

The template properties of left-handed synthetic polymers, the Z* form of poly[d(G-C)] and the Z form of poly[d(G-m5C)], have been investigated using an eucaryotic RNA polymerase, the class II enzyme from wheat germ. Results from a comparative kinetic study of transcription using the polynucleotide substrates in the B and Z conformations are reported. Optimal conditions for enzyme activity compatible with the preservation of the desired template conformation were determined. On the basis of several criteria, both physical (c.d. spectra of the polymers, sedimentability of the Z* form) and biochemical, it was demonstrated that the left-handed conformations of poly[d(G-C)] and poly[d(G-m5C)] serve as templates for wheat germ RNA polymerase II. The level of incorporation was less than that exhibited by the B form of poly[d(G-C)], the relative activity being a function of the precise experimental conditions. Activity ratios (Z*/B or Z/B) ranged from 0.1 to 0.5. The effect of various incubation parameters, including pH, salt concentration, temperature, and the presence of dinucleoside monophosphate primers were investigated. The Km values for nucleoside triphosphate substrates were slightly smaller for the Z* form of poly[d(G-C)] than for the B conformation. Titration of DNA (Z* or B) with enzyme and reciprocal experiments suggested that the reduced activity of left-handed templates might derive from the availability of fewer and/or lower affinity sites for initiation and/or translocation on these templates. Specific antibodies raised against left-handed DNA strongly inhibited the observed transcription of Z* and Z DNAs by wheat germ RNA polymerase II.

Ammonium Sulfate↗
PubMed1983

High-performance liquid chromatographic analysis of MTX, 7-OH-MTX and MTX derivatives: application to intracellular metabolism in tumor cells (HT 29).

In order to study the intracellular metabolism of Methotrexate (MTX) and the cytotoxicity of the antifolates, a specific paired-ion HPLC method has been developed which permits the simultaneous determination of DAMPA, MTX, 7-OH-MTX, MTX-G1, MTX-G2 and MTX-G3. Cells were incubated with 3H-MTX. The MTX metabolites were extracted, purified on SEP-PAK cartridges and further analysed by high-performance liquid chromatography (HPLC). The stationary phase was constituted by a C18 muBondapak and the mobile phase by 5 mM phosphate buffer (pH 7.4) containing 2.5 mM tetrabutylammonium nitrate. The elution was performed with a linear methanol gradient (20--30%). HPLC fractions were collected and radioactivity evaluated by beta counting (retention times: DAMPA = 12.93 min; MTX = 18.29 min; 7-OH-MTX = 21.13 min; MTX-G1 = 22.69 min; MTX-G2 = 26.81 min; MTX-G3 = 30.61 min). This analytical procedure was applied to separate and characterize multiple forms of MTX polyglutamate derivatives in HT 29, a human adenocarcinoma cell line varying the incubation time (4--18 h) and MTX concentration (0.6--10.6 micrometer). The incorporation process seems to be characteristic of a cell line resistant to MTX. The incorporation were very low and after a 4-h exposure time only 5% of the MTX was converted to polyglutamates. Between 1.6 and 10.6 microM MTX, no difference was observed in the polyglutamization. The defect in the incorporation of the drug and in the metabolization process in vitro could partially explain the failure of the MTX treatment in colorectal cancer.

Adenocarcinoma↗
PubMed1982

Non-processive transcription of poly[d(A-T)] by wheat germ RNA polymerase II.

RNA product distribution obtained during the transcription of poly[d(A-T)] by wheat germ RNA polymerase IIA under various experimental conditions was analyzed by high resolution polyacrylamide gel electrophoresis. Poly[r(A-U)] synthesis proceeded as if wheat germ RNA polymerase II was a non-processive enzyme: a ladder of RNA products of increasing lengths was obtained, which apparently, terminated at every other nucleotide. RNA release was not dependent upon nucleoside triphosphate substrate concentrations. A likely explanation would be that ternary complexes enzyme: DNA: RNA were very much unstable; moreover, oligonucleotides released were not re-used for further elongation by the enzyme.

DNA-Directed RNA Polymerases↗
PubMed1982

Metabolic type of muscles of the sheep. III. evolution with age and influence of sex.

Myofibrillar ATPase activity (in the presence of Ca2+ and Mg2+), activities of various glycolytic and mitochondrial enzymes, and haemin iron, glycogen, ATP and lactic acid levels were determined in the heart and five skeletal muscles of the sheep as a function of age and sex. During growth from 2 to 13 months, ATPase activity remained stable whereas most mitochondrial and glycolytic activities tended to decrease, particularly between 2 and 6 months. The isoenzyme composition of lactic dehydrogenase evolved markedly, with in particular a rise in the percentage of M4. Haemin iron level rose from 6 months. These modifications occurred in most of the skeletal muscles but rarely in the heart. Glycogen and ATP decreased progressively between 2 and 13 months. Overall, results indicated a regression of the metabolic differentiation among muscles between 2 and 13 months, particularly marked before 6 months. At 3 and 7 months, sex had practically no influence on the enzyme activities studied.

Adenosine Triphosphatases↗