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Biomedical subjects

D Perrin

Publications and source records attributed to D Perrin.

At least 73 records · Page 4Linked to original sources

Technical and anatomic factors in filling of distal portion of internal pudendal artery during arteriography.

Left and right internal pudendal arteriography was performed in 9 nonimpotent male patients (mean age 29 years) who had normal erections. Filling of distal branches of the internal pudendal artery and inter-pudendal anastomoses was improved by superselective catheterization of the anterior trunk of the internal iliac artery or an ischiopudendal trunk, and by the choice of hyperosmolar contrast medium. In the 5 cases where these conditions were fulfilled, filling of the distal branches of internal pudendal arteries was always symmetrical, or showed bifid artery with bilateral distribution on one side, except for the dorsal artery of the penis which appeared to be single in 2 cases. Interdependence between the two internal pudendal arteries was noted in 7 of 9 patients, through either direct anastomosis (7 cases) or bilaterally distributed deep artery of penis (2 cases).

Adult↗

Ventilatory predictors of pulmonary hypoplasia in congenital diaphragmatic hernia, confirmed by morphologic assessment.

We carried out a prospective study in 66 infants with congenital diaphragmatic hernia within the first 6 hours of life to determine whether outcome is related to the degree of underlying pulmonary hypoplasia, as predicted by preoperative PaCO2, when correlated with an index of ventilation (VI = mean airway pressure X respiratory rate) and confirmed by postmortem analysis of the lung. Those infants with PaCO2 greater than 40 mm Hg before surgery had a 77% mortality; when PaCO2 reduction could be achieved only with VI greater than 1000, the mortality was still greater than 50%. After repair, however, the ability to hyperventilate to PaCO2 less than 40 mm Hg proved to be an important determinant of survival; only one of 31 infants in this group died, whereas only two of 27 infants with PaCO2 greater than 40 mm Hg survived. In 16 infants with PaCO2 greater than 40 mm Hg despite hyperventilation, high-frequency oscillatory ventilation was started. This resulted in a rapid fall in PaCO2, but 14 of the 16 infants had only temporary improvement in oxygenation, and died. In five of the infants who died, alveolar number was assessed by postmortem morphometric analysis; there was a severe reduction to less than 10% of published normal neonatal values. Pulmonary vascular changes of increased muscularization were less remarkable than those observed in infants with persistent pulmonary hypertension. Our findings suggest that the degree of pulmonary hypoplasia (which would not be influenced by surgical repair), rather than the pulmonary vascular abnormality, mainly determines survival. Consideration could therefore be given to an initial nonsurgical approach to congenital diaphragmatic hernia, with the expectation that pulmonary function might improve and pulmonary vascular resistance decrease.

Hernia, Diaphragmatic↗

Operator-constitutive mutations of the Escherichia coli metF gene.

The Escherichia coli metF gene codes for 5,10-methylene-tetrahydrofolate reductase, the enzyme that leads to the formation of N-methyltetrahydrofolate, supplying the methyl group of methionine. Transcription of metF, as well as most of the methionine genes, is repressed by the metJ gene product complexed with S-adenosylmethionine. A metF'-'lacZ gene fusion was used to isolate mutants that have altered expression from the metF promoter. The nucleotide sequences of the metF regulatory region from five such mutants were determined. The mutations were located in the region previously defined as the potential target of the methionine repressor by its similarity to other binding sites. The mutationally defined metF operator thus consists of a 40-base-pair-long region, with five 8-base-pair imperfect palindromes spanning the metF transcription start. The altered operators do not recognize the purified repressor in an in vitro transcription-translation system, although the repressor binds efficiently to the metF wild-type operator.

5,10-Methylenetetrahydrofolate Reductase (FADH2)↗

Palindromic units from E. coli as binding sites for a chromoid-associated protein.

Several hundred copies of a highly conserved extragenic palindromic sequence, 20-40 nucleotides long, exist along the chromosome of E. coli and S. typhimurium. These have been defined as palindromic units (PU) or repetitive extragenic palindromes (REP). No general function for PUs has been identified. In the present work, we provide data showing that a protein associated with a chromoid extract of E. coli protects PU DNA against exonuclease III digestion. This provides the first experimental evidence that PU constitutes binding sites for a chromoid-associated protein. This result supports the hypothesis that PUs could play a role in the structure of the bacterial chromoid.

Bacterial Proteins↗

malM, a new gene of the maltose regulon in Escherichia coli K12. I. malM is the last gene of the malK-lamB operon and encodes a periplasmic protein.

The structure and expression of the distal part of the malK-lamB operon in Escherichia coli was studied. DNA sequencing was performed as far as a HinfI restriction site located 1313 base-pairs downstream from gene lamB. The open reading frame, formerly called molA, which begins 245 base-pairs downstream from gene lamB, is longer than was initially thought, and was renamed malM. It could encode a protein of 306 amino acid residues. The complete malM open reading frame was cloned under control of the tac 12 promoter. In maxicells, the resulting plasmid permitted tac12-promoted synthesis of two polypeptides, encoded by gene malM, with apparent molecular weights of 37 X 10(3) and 34.5 X 10(3). We provide strong evidence that the 34.5 X 10(3) Mr protein is derived from the 37 X 10(3) Mr protein by processing at the amino-terminal end, and that this processed form is located in the periplasmic space. We show that the chromosomal malM gene is expressed as part of the malK-lamB operon, and that its product is periplasmic. Finally, we demonstrate with nuclease S1 mapping experiments that the mRNA terminates at a typical rho-independent terminator located about 45 base-pairs beyond the end of gene malM, which is thus the last gene of the malK-lamB operon.

Amino Acid Sequence↗

Interactions of the Escherichia coli methionine repressor with the metF operator and with its corepressor, S-adenosylmethionine.

The metJ gene encoding the methionine aporepressor was placed under the control of a strong and inducible promoter, ptac. Bacterial strains carrying the recombinant plasmid pIP35 overproduced the regulatory protein by a factor of 200 over the wild type strain as determined by the immunoblot technique. The purified metJ gene product negatively controls the expression of the metF gene, in a cell-free system as shown by repression of beta-galactosidase synthesis under the control of the metF promoter. The metJ protein binds to a DNA fragment containing the potential operator of the metF gene with an affinity which is 10 times greater in the presence of S-adenosylmethionine than in its absence. Equilibrium dialysis experiments showed that the met aporepressor binds 2 mol of S-adenosylmethionine per mol of dimer with a dissociation constant of 200 microM.

Escherichia coli↗

A pH-induced increase in hydrophobicity as a possible step in the penetration of colicin E3 through bacterial membranes.

Exposure to low pH triggers an increase in the hydrophobicity of the colicin E3 molecule. Using a [3H] Triton X-100 binding assay we have shown that the amount of detergent (at supramicellar concentrations) associated with colicin E3 increased dramatically at pH 3.8 and below. Interaction of colicin E3 with asolectin vesicles was monitored by following its cross-linking with two different photoactivatable radioactive phospholipid analogues. At neutral pH colicin E3 was cross-linked with the phospholipid probing the membrane surface whereas at pH 4.5 and below, the bacteriocin reacted with the phospholipid probing the hydrophobic core of the bilayer. With the use of phase partitioning of proteins in Triton X-114 it was shown that at acidic pH whole colicin E3 and its immunity protein segregated in the detergent phase. After trypsin digestion of the colicin-immunity complex, the N-terminal portion of E3 (T1) and the immunity partitioned in the detergent phase at low pH. In contrast, the enzymic domain of the colicin (T2) remained in the aqueous phase and was recovered in a highly active form as a consequence of its dissociation from the immunity protein. These results are discussed in relation to the mechanism of entry of colicin E3 into bacterial cells.

Carbon Radioisotopes↗

Alpha-fodrin in the adrenal gland: localization by immunoelectron microscopy.

Localization of fodrin, the brain equivalent of spectrin (a protein constituent of the erythrocyte membrane cytoskeleton), was investigated at the ultrastructural level in rat adrenal gland. By use of an affinity purified antibody directed against the alpha-fodrin subunit, all chromaffin cells, cortical cells, nerve fibers, and their surrounding Schwann cells were found to be labeled close to the cytoplasmic side of their plasma membranes. The labeling appeared more intense for chromaffin cells, and secretory granules and mitochondria were frequently found to be associated with the zone containing alpha-fodrin in these cells. The immunostained zone was estimated to extend 230 +/- 70 nm into the cytoplasm. This localization is discussed in terms of what is known of the properties of spectrin, and possible roles of the molecule in the chromaffin cell are suggested.

Adrenal Glands↗

Clinical assessment and plasma pharmacokinetics associated with intramuscular or extradural alfentanil.

Patients with postoperative pain received alfentanil in doses of 15 micrograms kg-1 (n = 6) or 30 micrograms kg-1 (n = 6) via the extradural route or 15 micrograms kg-1 (n = 6) i.m. Effective analgesia was obtained in all patients in the extradural groups and in none in the i.m. group. Maximum pain relief developed within 10-15 min and the average duration of adequate analgesia was 78 min with the lower dose and 45 min with the higher dose, but this difference was not statistically significant. The 15-micrograms kg-1 extradural and i.m. groups showed the same pharmacokinetic pattern. Plasma concentrations were greater in the 30-micrograms kg-1 extradural group and this was associated with unwanted systemic effects. We conclude that the extradural administration of alfentanil can produce effective spinal analgesia. Its lipophilic property reduces the onset time, but does not modify its duration of action or impair absorption.

Adult↗

Chromaffin granule membrane-F-actin interactions and spectrin-like protein of subcellular organelles: a possible relationship.

The membrane of chromaffin granule, the secretory vesicle of adrenal medullary cells storing catecholamines, enkephalins, and many other components, interacts with F-actin. Using low shear falling ball viscometry to estimate actin binding to membranes, we demonstrated that mitochondrial and plasma membranes from chromaffin cells also provoked large increases in viscosity of F-actin solutions. Mitochondrial membranes also had the capacity to cause complete gelation of F-actin. In addition, vasopressin-containing granules from neurohypophysial tissue were shown to bind F-actin and to increase the viscosity of F-actin solutions. Using an antibody directed against human erythrocyte spectrin, it was found that a spectrin-like protein was associated with secretory granule membrane, mitochondrial membrane, and plasma membrane. The chromaffin granule membrane-associated spectrin-like protein faces the cytoplasmic side, is composed of two subunits (240 kD and 235kD ), the alpha-subunit (240 kD, pHi5 .5) being recognized by the antibody. Nonionic detergents such as Triton X-100 or Nonidet P40 failed to release fully active spectrin-like protein. In contrast, Kyro EOB , a different nonionic detergent, was found to release spectrin-like protein while keeping intact F-actin binding capacity, at least below 0.5% Kyro EOB concentration. Chromaffin cells in culture were stained with antispectrin antibody, showing the presence of spectrin-like protein in the cell periphery close to the cell membrane but also in the cytoplasm. We conclude that in living cells the interaction of F-actin with chromaffin granule membrane spectrin observed in vitro is important in controlling the potential function of secretory vesicles.

Acetylcholinesterase↗

[Populations of circulating T lymphocytes in patients with alcoholic cirrhosis].

The characteristics, the origin and the role in the determinism and/or in the course of hepatocellular necrosis of immune disorders in patients with alcoholic liver cirrhosis still remain unclear. In this study, the T-lymphocyte population was determined by sheep-rosette formation and labelling with OKT3-monoclonal antibodies. T-cell subsets were investigated using OKT4 and OKT8 monoclonal antibodies in the peripheral blood of 40 alcoholic patients with cirrhosis (CA) and of 23 non-cirrhotic alcoholic (ANC) patients. The results were compared to those in 34 healthy volunteers. A decrease in the percentage of T-lymphocytes in the two groups of alcoholic subjects was noted. However, this decrease was significant only in the CA group and only when OKT3 antibodies were used as markers. The lymphocyte-subset identified by OKT4 antibodies was decreased as well in the CA and ANC groups; however the OKT8+ subset was significantly decreased in the CA group only (p less than 0.001). No correlation was found between this decrease in OKT8+ subpopulation or between the increased OKT4/OKT8 ratio and all biochemical parameters of hepatic function measured in this study. The significance of this T-cell imbalance in CA has still to be elucidated, concerning both the functional activity of the reduced pool of OKT8+ cells (suppressor or cytotoxic?) and the mechanism of this decrease.

Adult↗

Opioid peptide effects on insulin release and c-AMP in islets of Langerhans.

The time course and specificity of the effect of opioid peptides on c-AMP production in the islets of Langerhans was examined. An enkephalin analogue, d-Ala2Me Phe4 Met(O)-ol enkephalin (DAMME, Sandoz) produced a significant stimulation of basal c-AMP levels, with a peak of stimulation at 5 minutes and a decline thereafter. These changes in intracellular c-AMP levels were of the same order of magnitude as those induced by other secretagogues, but did not coincide in time with the more rapid peak of enkephalin-induced insulin release. The rise in islet c-AMP and insulin secretion induced by DAMME and alpha-endorphin but not leu enkephalin was antagonised by naloxone. The effects of high and low concentrations of a variety of opioid peptides and naloxone on insulin release and islet c-AMP levels were determined, alpha-endorphin, dynorphin, leu enkephalin and met enkephalin all stimulated insulin secretion significantly, though not to the same extent. Higher concentrations of alpha-endorphin, dynorphin and met enkephalin inhibited insulin release relative to effects at low opiate concentrations. However, higher concentrations of leu enkephalin stimulated insulin release further. We conclude from these results that the mode of action of opioid peptides in stimulating insulin release is not via increased islet c-AMP exclusively. Furthermore, the results obtained with different classes of opioid suggest the presence of distinctive types of opiate receptor in islets of Langerhans.

Animals↗