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Biomedical subjects

D Doucet

Publications and source records attributed to D Doucet.

At least 19 recordsLinked to original sources

Foaming and interfacial properties of hydrolyzed beta-lactoglobulin.

beta-lactoglobulin (beta-lg) was hydrolyzed with three different proteases and subsequently evaluated for its foaming potential. Foam yield stress (tau0) was the primary variable of interest. Two heat treatments designed to inactivate the enzymes, 75 degrees C/30 min and 90 degrees C/15 min, were also investigated for their effects on foam tau0. Adsorption rates and dilatational rheological tests at a model air/water interface aided data interpretation. All unheated hydrolysates improved foam tau0 as compared to unhydrolyzed beta-lg, with those of pepsin and Alcalase 2.4L(R) being superior to trypsin. Heat inactivation negatively impacted foam tau0, although heating at 75 degrees C/30 min better preserved this parameter than heating at 90 degrees C/15 min. All hydrolysates adsorbed more rapidly at the air/water interface than unhydrolyzed beta-lg, as evidenced by their capacity to lower the interfacial tension. A previously observed relationship between interfacial dilatational elasticity (E') and tau0 was generally confirmed for these hydrolysates. Additionally, the three hydrolysates imparting the highest tau0 not only had high values of E' (approximately twice that of unhydrolyzed beta-lg), they also had very low phase angles (essentially zero). This highly elastic interfacial state is presumed to improve foam tau0 indirectly by improving foam stability and directly by imparting resistance to interfacial deformation.

Animals↗

Analysis of antifreeze proteins within spruce budworm sister species.

Spruce budworm (Choristoneura) species survive sub-zero winter temperatures by producing antifreeze proteins (AFPs) encoded by a multigene family of short and long isoforms. We report in this study the first analysis of antifreeze proteins from related Choristoneura sister species. The additional thirty amino acid insert found in the longer AFP isoforms maintains the proteins beta-helix and original fifteen amino acid (Thr-X-Thr) repeat motif. Analysis of the beta-helix region shows more divergent residues surround the conserved Thr residues. Maintaining the beta-helix structure and conserved Thr residues appear to be paramount for AFP function and surviving sub-zero winter temperatures. Two other species within the same lepidopteran clade, Ditrysia, do not appear to contain any AFP-like sequences.

Amino Acid Sequence↗

Structure-function relationships in spruce budworm antifreeze protein revealed by isoform diversity.

The spruce budworm, Choristoneura fumiferana, produces antifreeze protein (AFP) to assist in the protection of the overwintering larval stage. AFPs are thought to lower the freezing point of the hemolymph, noncolligatively, by interaction with the surface of ice crystals. Previously, we had identified a cDNA encoding a 9-kDa AFP with 10-30 times the thermal hysteresis activity, on a molar basis, than that shown by fish AFPs. To identify important residues for ice interaction and to investigate the basis for the hyperactivity of the insect AFPs, six new spruce budworm AFP cDNA isoforms were isolated and sequenced. They differ in amino-acid identity as much as 36% from the originally characterized AFP and can be divided into three classes according to the length of their 3' untranslated regions (UTRs). The new isoforms have at least five putative 'Thr-X-Thr' ice-binding motifs and three of the new isoforms encode larger, 12-kDa proteins. These appear to be a result of a 30 amino-acid insertion bearing two additional ice-binding motifs spaced 15 residues apart. Molecular modeling, based on the NMR structure of a short isoform, suggests that the insertion folds into two additional beta-helix loops with their Thr-X-Thr motifs in perfect alignment with the others. The first Thr of the motifs are often substituted by Val, Ile or Arg and a recombinantly expressed isoform with both Val and Arg substitutions, showed wild-type thermal hysteresis activity. The analysis of these AFP isoforms suggests therefore that specific substitutions at the first Thr in the ice binding motif can be tolerated, and have no discernible effect on activity, but the second Thr appears to be conserved. The second Thr is thus likely important for the dynamics of initial ice contact and interaction by these hyperactive antifreezes.

Amino Acid Motifs↗

A polydnavirus from the spruce budworm parasitoid, Tranosema rostrale (Ichneumonidae).

The calyx epithelium of the campoplegine wasp, Tranosema rostrale, contains typical ichneumonid polydnaviruses (PVs) that display an apparently uncommon association with the egg chorion. The latter structure features fine hair-like projections, longest around the egg's apices. In the lumen of the ovary, T. rostrale virus becomes lodged between these projections and forms a particulate coat around the egg. In the host, Choristoneura fumiferana, projections and associated virions are observed in close contact with basement membranes of fat body and muscle tissues, to which the eggs rapidly become attached following introduction into the host hemocoel. We discuss the implications of this unusual virus-chorion association in terms of immune protection, delivery of virus to specific host tissues, and the evolution of PVs.

Animals↗

The antifreeze potential of the spruce budworm thermal hysteresis protein.

Antifreeze proteins (AFP) inhibit ice growth by surface adsorption that results in a depression of the freezing point below the melting point. The maximum level of this thermal hysteresis shown by the four structurally unrelated fish AFP is approximately 1.5 degrees C. In contrast, hemolymph and crude extracts from insects can have 5 degrees to 10 degrees C of thermal hysteresis. Based on the isolation, cloning, and expression of a thermal hysteresis protein (THP) from spruce budworm (Choristoneura fumiferana), the vastly greater activity is attributable to a 9 kDa protein. This novel, threonine- and cysteine-rich THP has striking effects on ice crystal morphology, both before and during freezing. It is also 10 to 30 times more active than any known fish AFP, offering the prospect of superior antifreeze properties in cryoprotective applications.

Amino Acid Sequence↗

Toxicologic profile of iobitridol, a new nonionic low-osmolality contrast medium.

PURPOSE: The toxicologic profile of iobitridol, a new nonionic low-osomolality contrast medium, was evaluated in compliance with the current regulatory requirements in Europe, the USA and Canada. MATERIAL AND METHODS: The toxicity of iobitridol was tested following acute or repeated i.v. administration in several different species (mouse, rat, dog); single oral administration in the mouse and intracisternal injection in the rat. Furthermore, teratogenicity and mutagenicity were evaluated in the rat and rabbit. Local perivenous toxicity was assessed in the rabbit. RESULTS: The acute toxicity of iobitridol in the mouse is equivalent to that of iohexol, a reference product tested under the same conditions. Chronic administration (daily injections i.v. injection over 4 weeks) in the rat and dog did not demonstrate any particular toxicity for iobitridol. It should be noted that, unlike iohexol, iobitridol did not provoke any vacuolization of the renal tubular cells in the rat following repeated injections. Furthermore, this contrast agent did not show any teratogenic or mutagenic potential. The typical local inflammatory signs observed following perivenous injection in the rabbit were low in intensity and reversible. CONCLUSION: The toxicologic profile of iobitridol appears to be favorable and does not show any particular risk for clinical use under the usual indications of water soluble iodinated contrast agents.

Administration, Oral↗

Pharmacologic profile of iobitridol, a nonionic iodinated contrast medium.

The present article combines and summarizes the preclinic studies carried out in vitro and in vivo to determine the pharmacologic and biochemical profile of iobitridol, a new nonionic iodinated low-osmolality contrast medium (CM). The effects of this product on the main hemodynamic, bronchopulmonary, neurologic, renal, blood chemistry and electrophysiologic parameters and RBC morphology were studied in detail in comparison with CM in the same chemical category or with reference substances of the same osmolality. The in vivo studies were performed under conditions resembling clinical use. Iobitridol showed an excellent pharmacologic and biochemical profile, which was identical or superior to that of other products in its category.

Animals↗

[Electrocorticographic evaluation of the neurologic tolerability of iobitridol (Xenetix), a new non-ionic contrast medium in rabbits].

Electro-corticographic tracings (two longitudinal leads, bipolar assembly) were recorded from curarized rabbits (5/group) receiving selective internal carotid artery injections of either iobitridol, a new non-ionic contrast medium, or iohexol, the high osmolar diatrizoate or hypertonic mannitol (isotonic to the non-ionic agents). A further group was submitted to the surgical preparation but was not injected. The solutions were injected at a dose of 2.5 ml during 30 seconds. The animals were anaesthetized (halothane) during the surgical period. The permeability of the blood-brain barrier was assessed by means of the extravasation of Evans'blue. Tracings were visually assessed and a semi-quantitative method for blind evaluation of fast/slow rhythms was used. This method was pharmacologically validated by the use of pentobarbital and pentylenetetrazole. Diatrizoate and iohexol induced respectively 3 and 2 paroxystic tracings during or immediately after the injection period. Iobitridol and hypertonic mannitol did not cause such effect. Paroxystic tracings in the iohexol group were not associated with extravasation of Evans'blue in the cerebral parenchyma. Blood brain barrier was disrupted in all rabbits receiving the high osmolar agent diatrizoate. Tracings of the control group were characterized by a progressive increase of fast rhythms, as those of the iobitridol and mannitol groups. On the contrary, iohexol and especially diatrizoate induced an increase in the proportion of slow waves. Taken together, these data suggest that iobitridol shows an excellent tolerability potential for clinical use.

Animals↗

Modulation of the renal effects of contrast media by endothelium-derived nitric oxide in the rat.

RATIONALE AND OBJECTIVES: A possible involvement of endothelium derived relaxing nitric oxide (NO) in the pathogenesis of iodinated contrast media (CM)-induced nephrotoxicity was investigated in the rat. METHODS: Male rats (6 to 12 per group) were uninephrectomized. Six days later, the aorta was clamped above the renal artery and a low-osmolar contrast medium (CM), ioxaglate, was injected (1 mL/min; 3 minutes) via an aortic puncture in the single remaining kidney. Contrast medium was injected with or without the NO-synthase inhibitor L-NAME (100 mg/kg intravenously [i.v.] 5 minutes before CM). One group received L-Arginine, the physiological precursor of NO (100 mg/kg i.v.), 5 minutes before L-NAME. Phenylephrine (300 micrograms/kg; 30 min) was used as a vasoconstrictive NO-independent control. The effects of iohexol, another low-osmolar CM, on creatinine clearance (CrCl) were also studied with and without pretreatment with L-NAME. A control group was subjected to a 3-minute renal ischemia only. Creatinine clearance and urinary N-acetyl-beta-D-glucosaminidase (NAG) excretion were determined before, and 24 and 48 hours after CM administration. Blinded histologic analysis was carried out after completion of the study. RESULTS: When administered alone, neither L-NAME nor L-arginine modified CrCl. Ioxaglate mildly but significantly decreased CrCl at 24 hours (-26.5% of preinjection value). This was similar to the effect observed in the control group subjected to ischemia only. When associated with L-NAME, ioxaglate markedly decreased CrCl (-58 + 11% at 24 hours, P < .05 vs. ioxaglate alone). A similar interaction was noted in the case of iohexol. L-NAME also markedly increased ioxaglate-induced urinary NAG excretion. Phenylephrine had a similar impact on renal function. L-arginine pretreatment reduced the increase in serum creatinine induced by L-NAME+ioxaglate (68 + 17 mumol/L vs. 175 + 59 mumol/L for L-NAME+ioxaglate; P < .05) and urinary NAG excretion. Ioxaglate alone induced only tubular epithelial vacuolization. When associated with L-NAME, this CM induced tubular and vascular lesions, as well as necrosis in the outer medulla. Such histologic effects were clearly inhibited by L-arginine. CONCLUSION: These data indicate that L-NAME, a specific inhibitor of NO-synthase, and phenylephrine, accentuate the nephrotoxicity of CM in the rat. This is consistent with results from the literature showing that CM-toxicity is enhanced by renal ischemia.

Acetylglucosaminidase↗

Pharmacokinetic study of gadolinium-DOTA in control and streptozocin diabetic rats.

The pharmacokinetics of gadolinium tetraazacyclododecanetetraacetic acid (Gd-DOTA), a contrast agent used in magnetic resonance imaging, have been evaluated in control and streptozocin-diabetic rats of different ages. In control rats, an age-related decrease in the Gd-DOTA elimination rate was noted, supported by a significantly lower apparent total body clearance and a significantly higher mean residence time. In diabetic rats, a similar but less important age-related change in the mean residence time and the apparent total body clearance was observed. Regardless of age-related differences in the pharmacokinetic parameters, a diabetic state induced several alterations in the Gd-DOTA pharmacokinetic parameters. The apparent total body clearance was significantly higher and the mean residence time significantly lower in diabetic rats indicating a higher elimination rate of Gd-DOTA. An important age-related increase in the volume of distribution at steady-state was noted in diabetic rats.

Animals↗

[Pharmacotoxicological profile of a non-ionic LOCM at high hydrophilicity: ioversol].

The authors summarize the results of the main preclinical studies conducted to evaluate the tolerance of ioversol (Optiray), a new non ionic Low Osmolality Contrast Medium (LOCM). The various studies conducted by different routes of administration, in a number of animal species and at different dosages led to the conclusion that the product was equivalent or superior to the other non ionic agents used as references. This high inertia towards the biological systems particularly well evidenced in the neurologic tolerance studies has to be relied to the non ionic LOCM character of the agent but also to its highest hydrophilicity in its class and to the good distribution of this hydrophilicity around the molecule. These physiochemical characteristics which differentiate ioversol from the other products of its class make it a promising product for clinical use in humans.

Animals↗