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Biomedical subjects

E Quentin

Publications and source records attributed to E Quentin.

At least 19 recordsLinked to original sources

An integrated study of threonine-pathway enzyme kinetics in Escherichia coli.

We have determined the kinetic parameters of the individual steps of the threonine pathway from aspartate in Escherichia coli under a single set of experimental conditions chosen to be physiologically relevant. Our aim was to summarize the kinetic behaviour of each enzyme in a single tractable equation that takes into account the effect of the products as competitive inhibitors of the substrates in the forward reaction and also, when appropriate (e.g. near-equilibrium reactions), as substrates of the reverse reactions. Co-operative feedback inhibition by threonine and lysine was also included as necessary. We derived the simplest rate equations that describe the salient features of the enzymes in the physiological range of metabolite concentrations in order to incorporate them ultimately into a complete model of the threonine pathway, able to predict quantitatively the behaviour of the pathway under natural or engineered conditions.

Aspartate Kinase↗

Temperature dependence of the coupling efficiency of rat liver oxidative phosphorylation: role of adenine nucleotide translocator.

We have reinvestigated the temperature dependence of the coupling efficiency of energy conversion in isolated rat liver mitochondria. We observed that respiratory control increased with temperature. Moreover, in the same conditions, the ATP/O ratio increased. The measurement of the control coefficients of adenine nucleotide translocator on respiratory and ATP synthesis rates showed that at 28 degrees C, this translocator exerted the same control (about 0.5) on both fluxes. At 4 degrees C, it no longer exerted control on respiratory flux when its control on ATP synthesis flux came close to 1. In addition, ATP/O ratio values and control coefficients on ATP synthesis flux were bound by a unique linear relationship irrespective of temperature. In conclusion, the decrease in ATP/O ratio with temperature is a direct consequence of an increase in the kinetic control exerted by the adenine nucleotide translocator on ATP synthesis.

Adenosine Diphosphate↗

A novel large dermatan sulfate proteoglycan from human fibroblasts.

Human skin fibroblasts express, in addition to versican, a second large chondroitin sulfate/dermatan sulfate proteoglycan, which has been investigated with the aid of a specific antiserum in cultures of fetal fibroblasts. Its core protein, obtained after chondroitin ABC lyase treatment, exhibits an apparent molecular mass of about 740 kDa in the absence of a reducing agent whereas reduction produces two core proteins of 460 and 300 kDa, respectively. Both subunits carry one or very few dermatan sulfate chains of about 20 kDa which are of similar chemical composition irrespective of the type of subunits to which they are attached. Tryptic peptide maps of [35S]methionine-labeled core proteins indicated that both subunits are related neither to each other nor to versican, suggesting that the proteoglycan exists predominantly as a heterodimeric molecule. It is insensitive to collagenase and does not interact with hyaluronan. Pulse-chase experiments suggested that the core proteins are different gene products. Dimerization begins soon after core protein synthesis but requires more than 2 h for completion. Glycosaminoglycan synthesis occurs immediately prior to secretion. A small proportion of both subunits may be secreted in form of a monomeric proteoglycan. The heterodimeric proteoglycan is a major proteoglycan species of fetal fibroblasts. The secreted product represents 10-20% of [35S]methionine and about 5-10% of [35S]sulfate incorporated into secreted proteoglycans.

Cells, Cultured↗

Elevated plasma levels of the immunosuppressive complement fragment Ba in renal failure.

The complement fragment Ba is a 33 kD activation product of factor B which suppresses human B-lymphocyte functions in vitro. We report that plasma levels of Ba are highly elevated in patients with chronic renal failure (4.84 +/- 3.58 micrograms/ml) and in patients with end-stage renal disease undergoing regular hemodialysis (16.1 +/- 6.1 micrograms/ml) as compared to normals (1.01 +/- 0.30 micrograms/ml). Ba levels were strictly correlated with the creatinine clearance. The urinary excretion of Ba was 165-fold higher in patients with tubular proteinuria than in normals. These results indicate that the kidney is the major catabolic site for Ba. In addition, direct evidence was obtained for an enhanced turnover of the alternative pathway of complement in renal failure that, although it appears to be less important than the renal retention of Ba, contributes to elevated Ba plasma levels in these patients. Ba concentrations in dialysis patients who responded to hepatitis B vaccination were significantly lower than in non-responders. Furthermore, the in vitro IgM synthesis by purified mononuclear cells was negatively correlated with Ba concentrations determined in the plasma of these patients. These results suggest that the accumulation of Ba contributes to the defective immune response in patients with renal failure.

Adult↗

Determination of the molecular structure of the human free secretory component.

Here, we present the experimental data, leading to determination of the primary structure, the linkage of the carbohydrates and the arrangements of the disulfide bonds of the human free secretory component. Methods of protein chemistry were used. The protein can be divided into five homology regions and is a member of the immunoglobulin superfamily.

Amino Acid Sequence↗

A genetic defect in the biosynthesis of dermatan sulfate proteoglycan: galactosyltransferase I deficiency in fibroblasts from a patient with a progeroid syndrome.

A small proteoglycan that contains only a single dermatan sulfate chain is the main proteoglycan synthesized by skin fibroblasts. Fibroblasts from a patient with progeroidal appearance and symptoms of the Ehlers-Danlos syndrome have a reduced ability of converting the core protein of this proteoglycan into a mature glycosaminoglycan chain-bearing species. This abnormality is the consequence of a deficiency in galactosyltransferase I (xylosylprotein 4-beta-galactosyltransferase; EC 2.4.1.133), which catalyzes the second glycosyl transfer reaction in the assembly of the dermatan sulfate chain. The glycosaminoglycan-free core protein secreted by the patient's fibroblasts bears an unsubstituted xylose residue. The mutant enzyme is abnormally thermolabile. Preincubation of fibroblasts at 41 degrees C leads to a further reduction in the production of mature proteoglycan and affects the capacity for glycosaminoglycan synthesis on p-nitrophenyl beta-D-xyloside more strongly in the mutant than in control cells.

Carbohydrate Sequence↗

Complement activation in patients with renal failure as detected through the quantitation of fragments of the complement proteins C3, C5, and factor B.

Using sensitive and highly specific enzyme-linked immunosorbent assays fragments of the complement proteins C3, C5, and factor B were quantitated in patients with renal failure. During hemodialysis on new cuprophan membranes raised levels not only of C3a, but in addition of activated C3, C5a, and Ba were demonstrated. In patients with chronic renal failure and end-stage renal disease plasma concentrations of Ba and activated C3 were markedly elevated independent of hemodialysis. This finding is taken as an indication of a continuous recruitment of the alternative pathway of complement in these patients. As the detected complement protein fragments are known to exert immune regulatory functions these findings may imply that these peptides are involved in the maintenance of the immune suppressed state in renal failure.

Acute Kidney Injury↗

[Modification of atrial natriuretic peptide (ANP) and cyclic GMP by hemofiltration and hemodialysis].

ANP and c-GMP concentrations in 7 patients with chronic renal failure (CRF) undergoing regular hemofiltration (HF) were determined. After switching to hemodialysis (HD) under identical ultrafiltration and treatment time no significant difference of the ANP and c-GMP profiles was detected, suggesting that the type of treatment does not affect ANP and c-GMP plasma levels. In both procedures a continuous decrease of ANP and c-GMP was observed. Head down tilting to compensate hypotension during HD was immediately followed by an increase in ANP and c-GMP during ultrafiltration. An acute onset of tachyarrhythmia absoluta during HD was also accompanied by a rise in ANP plasma concentrations. This demonstrates that ANP secretion is not altered in patients with CRF. Since ANP plasma levels closely correlate with intravascular volume, periodic determination of this hormone in HD/HF patients may provide diagnostic information to detect volume overload.

Atrial Natriuretic Factor↗

[Loss of renal functional reserve following kidney transplantation and in patients with advanced disorders of liver function].

Renal functional reserve capacity was evaluated in healthy controls, kidney transplant recipients and patients with impaired liver function by simultaneous measurements of periodic clearances of inulin, PAH and creatinine every 30 minutes before, during and after infusion of an amino acid (AA) solution. During AA infusion glomerular filtration rate rose in 10 healthy controls to about 35% above basal values (inulin clearance from 107 +/- 6 to 144 +/- 7 ml/min, p less than or equal to 0.0005), renal plasma flow increased by 27% (PAH clearance from 530 +/- 25 to 675 +/- 40 ml/min, p less than or equal to 0.002). 8 renal transplant recipients with good and stable renal function (creatinine clearance above 65 ml/min) showed no rise in GFR and RPF, as did 10 patients with severe impairment of liver function and normal basal kidney function (creatinine clearance above 100 ml/min). The lack of renal functional reserve in kidney transplant recipients might indicate a hyperfiltration of the transplanted kidney. This could affect the longtime prognosis of these patients. The liver seems to play a role in the mediation of the amino acid-induced rise of GFR, supporting the hypothesis of a putative liver hormone regulating GFR after protein ingestion or AA infusion.

Adult↗

Glycosaminoglycan-free small proteoglycan core protein is secreted by fibroblasts from a patient with a syndrome resembling progeroid.

A male patient, 4 years 9 mo old and having progeroidal appearance, exhibited delayed mental development and multiple abnormalities of connective tissues including growth failure, osteopenia of all and dysplasia of some bones, defective deciduous teeth, loose but elastic skin, delayed wound healing with formation of thin atrophic scars, scanty scalp hair, hypotonic muscles, and hypermobile joints. Skin fibroblasts of the patient converted only about half of the core protein of the small proteodermatan sulfate to a mature glycosaminoglycan chain-bearing proteoglycan. The remaining core protein, which contained complex-type asparagine-bound oligosaccharides, was secreted with almost normal kinetics. Xylosyltransferase activity and the synthesis of other proteoglycan types were normal. Normal induction of glycosaminoglycan synthesis occurred in the presence of 1 mM, but there was very little induction in the presence of 0.01 mM p-nitrophenyl-beta-xyloside. An antibody against an N-terminal pentadecapeptide of the core protein recognized the glycosaminoglycan-free core protein from the patient less well than the chain-bearing protein treated with chondroitin ABC lyase. Though these results do not define the basic defect unambiguously, they provide the first report of a disorder being due to an abnormality in small proteoglycan biosynthesis.

Abnormalities, Multiple↗

Biosynthesis and processing of proteodermatan sulphate.

The biosynthesis and processing of the small iduronic acid-rich proteodermatan sulphate (PDS) was studied in cultured human skin fibroblasts and arterial smooth muscle cells (SMC) with the aid of core-directed antibodies and various inhibitors of protein synthesis, intracellular transport, and glycosylation. Components of the linkage region became attached to the core protein most likely in a pre-Golgi compartment. Phosphorylation of PDS precursors also occurred in the endoplasmic reticulum with a minor contribution by the Golgi complex. Serine residues and the linkage region were identified as phosphorylated species in secreted PDS. Blockade of transport by monensin did not affect 6-sulphation but affected uronic acid epimerization and 4-sulphation. On relief from the monensin block, additional sulphation along the glycosaminoglycan chain was possible, whereas chain elongation was as in the continuous presence of the drug. Asparagine-bound oligosaccharides or glycosaminoglycan chains were not required for secretion of PDS or core protein. PDS from fibroblast and SMC secretions differed markedly in the composition of the glycosaminoglycan chains. No significant difference, however, was found on isoelectric focusing of core protein and after limited proteolysis of chondroitin ABC lyase-treated core protein. Tryptic and chymotryptic peptide maps of iodinated core proteins were similar.

Arteries↗

[The primary structure of human free secretory component and the arrangement of disulfide bonds].

The amino-acid sequence and the arrangement of the disulfide bonds of the human free secretory component were completely elucidated by the methods of protein chemistry. The free secretory component is a monomeric glycoprotein (Mr approximately 86000), consisting of 558 amino acids with 7 carbohydrate chains bound to asparagine. The protein contains 20 cysteine residues but, as a special feature, no methionine. The polypeptide chain is divided into five regions of internal homology, 104 to 114 amino acids in length. The 20 cysteine residues form 10 disulfide bonds, 9 of which confirm the internal homology by their characteristic arrangement. The free secretory component also shows homology to immunoglobulins in some sections. A computer-supported tertiary structure is proposed for the free secretory component.

Amino Acid Sequence↗

[Renal cancer today - suspicion, diagnosis or therapy? (author's transl)].

The present-day situation of recording and diagnosis of malignant renal tumours was investigated on 50 patients recently seen by us. It was found that the complex radiological diagnosis (composed of excretion urography, isotope nephrography, renoscintigraphy, renovasography and additional investigations) is very well suited to show renal cancer in all stages taking into consideration localisation, extent and relations to surrounding tissue. As ever, the difficulty for a clarifying diagnosis still lies in seeing the patient as early as possible. This is due to the almost exclusively uncharacteristic symptoms which, additionally, very frequently coincide with symptoms of other diseases. The only chief symptom is clearly 'painless haematuria'. Relatively large renal tumours also escape palpation. The possibility of tracing haematurias during screening tests with 'Reagnost haemoglobin' tablets is discussed.

Aged↗