PubMed HealthSearch

Biomedical subjects

G Laroche

Publications and source records attributed to G Laroche.

At least 19 recordsLinked to original sources

Characterization by infrared spectroscopy of the interaction of a cardiotoxin with phosphatidic acid and with binary mixtures of phosphatidic acid and phosphatidylcholine.

The effect of cardiotoxin IIa from Naja mossambica mossambica, a small basic protein extracted from snake venom, on dimyristoylphosphatidic acid (DMPA) and on equimolar mixtures of DMPA and dimyristoylphosphatidylcholine (DMPC) has been studied by Fourier transform infrared spectroscopy. The interaction of cardiotoxin with DMPA dispersions decreases both the cooperativity of the phase transition of the lipid and the molecular order of the lipid acyl chains in the gel phase. This effect increases with the proportion of the toxin in the complexes and leads to the total abolition of the phase transition of DMPA at a lipid-to-protein molar ratio of 5. Small-angle X-ray results demonstrate that the structure of the lipid-protein complexes is poorly ordered and gives rise to broad diffusion peaks rather than to well-resolved diffraction patterns. Infrared spectra of oriented cardiotoxin-DMPA films show that the protein is not homogeneously oriented with respect to the bilayer surface. The destabilization of the gel-phase structure of DMPA by cardiotoxin also results in a deeper water penetration in the interfacial region of the lipid since more carbonyl ester groups appear to be hydrogen bonded in the presence of the toxin. The infrared results on the phosphate group vibrations also indicate clearly that the basic residues of cardiotoxin interact strongly with the phosphate group of DMPA that becomes partly ionized at a pH as low as 6.5. The results obtained on the interaction of cardiotoxin with an equimolar mixture of DMPA and DMPC clearly demonstrate the ability of this toxin to induce lateral phase separation in this mixture with one phase containing DMPA-rich domains perturbed by cardiotoxin while the second phase is composed of regions enriched in DMPC. Comparison of the results of the current study with those obtained on other basic proteins and polypeptides suggests that charge-induced phase separation occurs only when the charge density on certain regions of the protein structure is high enough to lead to efficient electrostatic interactions with anionic phospholipids. This condition occurs only when the conformation of the protein or polypeptide is well-ordered at the lipid interface.

Cobra Cardiotoxin Proteins

Structure and dynamics of dimyristoylphosphatidic acid/calcium complexes by 2H NMR, infrared, spectroscopies and small-angle x-ray diffraction.

The structural and dynamic properties of complexes of dimyristoylphosphatidic acid (DMPA) and calcium ions have been characterized by 2H NMR, Raman, and infrared spectroscopies and small-angle X-ray diffraction. All techniques used show that these complexes do not undergo a cooperative thermotropic phase transition. Small-angle X-ray diffraction unambiguously demonstrates that the structure of the lipid molecules of the DMPA/Ca2+ complexes remains lamellar even at a temperature as high as 85 degrees C. Raman results indicate that within this temperature range, only a few trans-gauche isomerizations of the C-C bonds of the phospholipid acyl chains arise in this system. The 2H NMR spectra indicate that the DMPA chains are highly motionally restricted up to 65 degrees C and that higher temperatures might activate some low-frequency overall motions of entire lamellar domains. Small-angle X-ray scattering and 2H NMR spectroscopy of 2H2O also show that the interaction of calcium with DMPA promotes an important dehydration of the lipid assembly, even though the latter technique clearly demonstrates that some water molecules remain strongly bond in the DMPA/Ca2+ complexes. The carbonyl stretching mode region of the infrared spectrum of DMPA/Ca2+ complexes suggests that these water molecules are trapped near the interfacial region of the lipid membrane and are hydrogen bonded with the carbonyl groups of the lipid. Finally, comparison of the phosphate stretching mode region of the infrared spectra of complexes of DMPA with calcium ions with those of model compounds provides strong evidence that calcium ions bind to both charges of the phosphate group of DMPA and form bridges between adjacent bilayers.

Glycerophospholipids

Bifidobacterium from fermented milks: survival during gastric transit.

Two Bifidobacterium strains contained in two different fermented milks behave very differently when exposed to an in vitro simulated gastric environment. One strain survives very well during at least 90 min (greater than 10(7)/g), but the second strain studied is much less resistant. These in vitro results, with slight differences, were confirmed by an in vivo study in humans. The assessment of the gastric emptying rate of these products allows an estimation of the amount of Bifidobacterium that may pass into the small intestine.

Adult

Coupled changes between lipid order and polypeptide conformation at the membrane surface. A 2H NMR and Raman study of polylysine-phosphatidic acid systems.

Thermotropism and segmental chain order parameters of sn-2-perdeuteriated dimyristoyl-phosphatidic acid (DMPA)-water dispersions, with and without poly(L-lysine) (PLL) of different molecular weights, have been investigated by solid-state deuterium NMR spectroscopy. The segmental chain order parameter profile of this negatively charged lipid is similar to that already found for other lipids. Addition of long PLL (MW = 200,000) increases the temperature, Tc, of the lipid gel-to-fluid phase transition, whereas short PLL (MW = 4000) has practically no effect on Tc. In the fluid phase both varieties of PLL increase the "plateau" character of segmental order parameters up to carbon position 10. At the same reduced temperature, long PLL more significantly increases the segmental ordering, especially at the methyl terminal position. This leads to the conclusion that polar head-group capping and charge neutralization by PLL induce severe changes in lipid chain ordering, even down to the bilayer core. The structure of PLL bound to the lipid bilayer surface was monitored by Raman spectroscopy, following the amide I bands. Results show that the lipid gel-to-fluid phase transition triggers a conformational transition from ordered beta-sheet to random structure of short PLL, while it does not affect the strongly stabilized beta-sheet structure of long PLL. It is concluded that both short and long PLL can efficiently cap and neutralize lipid head groups, whatever their structure, and that peptide length is a key parameter in whether lipids or peptides are the driving force in conformationally coupled changes of both partners in the membrane.

Chemical Phenomena

Expanded polytetrafluoroethylene prostheses as secondary blood access sites for hemodialysis: pathological findings in 29 excised grafts.

The popularity of expanded polytetrafluoroethylene (PTFE) arteriovenous fistulas for hemodialysis access is increasing. The low infection rate, low thrombogenicity and low body reactivity make it an important blood access for patients on routine hemodialysis. The authors examined 29 PTFE graft fistulas surgically excised from patients on hemodialysis for one or more of the following complications: infection 7, 2 with associated hemorrhage; thrombosis 9; aneurysm 11, 3 with associated thrombosis; stenosis 5, 3 with associated thrombosis; hemorrhage 4 and arterial steal 2. The explanted prostheses were reinforced Gore-tex in 20, Gore-tex thin wall in 1, Impra I in 4, Impra II in 3 and Vitagraft in 1. It was also noted that the implants failed in the short term mainly because of infection, in the medium term mainly because of thrombosis and stenosis, and in the long term usually because of aneurysm.

Adult

Biologic and structural evaluation of 80 surgically excised human umbilical vein grafts.

Human umbilical vein is a valuable alternative to autogenous saphenous vein in lower-limb revascularization. However, the long-term patency is not as good. To understand better the reasons for this, the authors studied the biologic and structural changes occurring in 80 segments of human umbilical vein (HUV) grafts excised from 70 patients. Morphologic, histologic and scanning electron microscopy examinations were performed on each specimen. It was noted that increasing duration of implantation was associated with greater encapsulation and less surrounding inflammatory reaction. Twenty-six percent of clinically noninfected grafts were found to harbour bacteria. Bacteremic colonization was often in the folds of the luminal surface. As a result of these findings, the authors emphasize the need for complete excision of clinically infected grafts. Anastomoses between HUV segments are discouraged because they are associated with a high frequency of infection and a corresponding decrease in duration of implantation. Delamination of the wall was common, and such sites may represent areas at risk for further degradation. Continued surveillance of the biologic and structural changes occurring in excised grafts remains an important method in increasing our understanding of the evolutive complications of HUV grafts.

Adult

Study of the effect of poly(L-lysine) on phosphatidic acid and phosphatidylcholine/phosphatidic acid bilayers by raman spectroscopy.

The effect of polylysine (PLL) on dimyristoylphosphatidic acid (DMPA), on dimyristoyl-phosphatidylcholine (DMPC), and on mixtures of these lipids was investigated by Raman spectroscopy. These results show that long polylysine (Mr approximately 200,000) increases the stability of the acyl chain matrix of DMPA to form a more closely packed structure with a stoichiometry of one lysine residue per PA molecule. On the other hand, short PLL (Mr 4000) destabilizes the PA bilayer, and the complex formed undergoes a gel to liquid-crystalline transition at a lower temperature than of the pure lipid. For both cases, we have observed that bound polylysine adopts a beta-sheet conformation as opposed to the alpha-helical structure previously found for dipalmitoylphosphatidylglycerol/long PLL complexes [Carrier, D., & Pézolet, M. (1984) Biophys. J. 46, 497-506]. The difference in the thermal behavior of complexes of DMPA with long and short polylysines is believed to be associated with the fact that in the complex the long polypeptide adopts the beta-sheet conformation over the whole range of temperatures investigated while the short one undergoes a change of conformation from beta-sheet of random coil upon heating. Therefore, the conformation of the lipid-bound polypeptides depends on the nature of the polar head group of the lipid, not only on its net charge, and it affects considerably the thermotropism of the lipid. On the other hand, both long and short polylysines show no affinity for phosphatidylcholine since the temperature profiles of DMPC and of DMPC/PLL complexes exhibit exactly the same behavior.(ABSTRACT TRUNCATED AT 250 WORDS)

Dimyristoylphosphatidylcholine

The virgin, modified, human, umbilical vein graft: morphologic characteristics and mechanical properties.

The authors used macrophotography, endoscopy, roentgenography, light microscopy, and transmission and scanning electron microscopy to assess the morphologic characteristics of 37 virgin, human, umbilical vein grafts. The specimens showed deep longitudinal folds (22%) and multiple transverse folds of the intimal surface (27%), irregularity of wall thickness (41%) and intimal breakdown exposing thrombogenic layers of the vein wall to blood (19%). The mechanical properties also studied were dynamic compliance, Young's modulus, breaking strength and breaking strain. The mean dynamic compliance was 6.44 X 10(-4) mm Hg-1 (SD = 2.02 X 10(-4) mm Hg-1, i.e., 31%), which is in agreement with results of others. The specimens showed great variability in Young's modulus, breaking strength and breaking strain. The structural deficiencies and variable mechanical properties of human umbilical vein grafts require investigation to determine their effect on patency rates due to increased thrombogenicity and thus to establish whether there are potential hazards associated with their use.

Biomechanical Phenomena