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Biomedical subjects

L Robert

Publications and source records attributed to L Robert.

At least 19 recordsLinked to original sources

Lack of postmortem digestion of tapeworms in Golden hamsters experimentally infected with Taenia solium.

Taenia solium causes human neurocysticercosis, a public health problem in Mexico and other developing countries. Surprisingly, tapeworm carriers are very rarely found and in necropsy studies practically no tapeworms have been reported. In this paper we analyze the possibility that, after the death of the host, tapeworms could easily be destroyed in the intestine. Our experiments, performed in the hamster model, suggest that the absence of tapeworms in human intestine during necropsy is not due to postmortem digestion.

Animals↗

The metabolic syndrome and the Maillard reaction. An introduction.

With the rapid increase of life expectancy in western societies, appeared also a new phenomenon, obesity, which took during the recent decades pandemic dimensions. One of the consequences was the appearance of type II diabetes in much younger persons than before. The result of intensive research in this field over the last decades led, among other achievements to the identification of biological and molecular symptoms which together were reclassified as the "metabolic syndrome". Questionable as far as its originality is concerned, it did however good service to practitioners by formulating criteria for diagnostic and therapeutical purposes. Among the underlying biochemical mechanisms the Maillard reaction, the non enzymatic glycosylation of proteins and nucleotide bases followed by the formation of advanced glycation and products (AGE-s) plays an important role. Several recent experimental results confirm this statement, some of them are published in this issue. The reviews and original contributions form together an up to date report on this important pathology and support again the importance of posttranslational and environmental factors influencing gene expression and cellular functions.

Aging↗

Effect of advanced glycation end-products on cell proliferation and cell death.

The effect of advanced glycation end products (AGE-s) was studied on the proliferation and cell death of human skin fibroblasts in culture. Several AGE-products were prepared from proteins, a peptide and amino acids, using Glucose or Fructose, with or without Fe2+. The AGE preparations increased cell death at the 7th day, after only 72 hours of incubation. Some of these glycation products modified also proliferation. This effect of AGE-s was even maintained without these products in fresh medium for a second period of incubation up to 10 days from the start of the experiment. In order to explore the role of AGE-receptors, especially of AGE-receptor and of growth factor receptors (fibroblast and epidermal growth factors receptors), antibodies to these receptors were added to cell cultures and their effect on both cell death and proliferation were determined as for the AGE-s. These anti-receptor antibodies imitated to some extent the results obtained with AGE-s, producing increase of cell death and proliferation, followed above a certain concentration of antibodies by a decrease and a new increase or plateau. This might correspond to the internalization of the receptors followed by a re-expression on the cell membrane. The role of receptor-mediated Reactive Oxygen Species-production was also explored using scavengers: N-acetyl-cysteine (NAC), L-Carnosine, superoxide dismutase (SOD) and Catalase. Several of these scavengers decreased cell death, suggesting that Reactive Oxygen Species-production is partially involved in the observed phenomena.

Antibody Formation↗

Pharmacological properties of rhamnose-rich polysaccharides, potential interest in age-dependent alterations of connectives tissues.

Rhamnose-rich oligo- and polysaccharides (RROPs) were tested for their potential pharmacological properties using human skin fibroblasts in serial cultures. The substances tested were shown to stimulate cell proliferation, decrease elastase-type activity, stimulate collagen biosynthesis, and protect hyaluronan against free radical mediated degradation. These reactions appear to be triggered by the mediation of a specific alpha-L-rhamnose recognizing lectin-site acting as a receptor, transmitting signals to the cell-interior. The rapid increase of intracellular free calcium after addition of RROP-1 and preliminary data using micro arrays appear also to confirm this contention.

Adult↗

Protection by rhamnose-rich polysaccharides against the cytotoxicity of Maillard reaction products.

Previous experiments have shown that AGE-products added to human skin fibroblast cultures increased the number of dead cells floating on top of the culture fluid and took up vital dye [1]. In these experiments, we tested several rhamnose-rich polysaccharides for protection against the cytotoxic effect of AGE-s. Added at relatively low concentrations (between 10 and 100 microg/ml) to the culture medium, several of the tested rhamnose-rich oligo- and polysaccharides (RROP-s) gave a significant protection against AGE-induced cytotoxicity. Their effect on cell proliferation was also tested. The number of cells at saturation density was also shown to be influenced by AGE-products added to the cultures. This effect was also, at least partially, corrected by the rhamnose-rich oligo- and polysaccharides. These substances might therefore be considered as of potential therapeutical interest against hyperglycemia induced cytotoxic effects as in type II-diabetes.

Cell Death↗

Sensitivity of a Lamb wave sensor with 2 microm AlN membrane.

Anti-symmetrical Lamb wave mode A0 presents a large sensitivity to mass loading and can be used in contact with liquids with a small attenuation. The advantages of this system are the possibility to get a large mass sensitivity. The sensitivity increases when the thickness of membrane decreases. Therefore the problem is to obtain thin piezoelectric membranes. A membrane of AlN with a thickness of 2 microm has been made. The measured mass sensitivity with a fluid is 200 cm(2) g(-1). In a practical use point of view, the problem in this kind of sensor is its temperature sensitivity. In order to reduce effective temperature sensitivity, a device with thin metallic strips is presented. On the same membrane two different waves with perpendicular propagating directions are produced. Experimentally, temperature sensitivity is rather different depending on the propagation direction but mass sensitivity is almost the same, this allows distinguishing temperature effects from those due to mass loading on the frequency shift measurements.

Acoustics↗

Evidence for complete surface wave band gap in a piezoelectric phononic crystal.

A complete surface acoustic wave band gap is found experimentally in a two-dimensional square-lattice piezoelectric phononic crystal etched in lithium niobate. Propagation in the phononic crystal is studied by direct generation and detection of surface waves using interdigital transducers. The complete band gap extends from 203 to 226 MHZ, in good agreement with theoretical predictions. Near the upper edge of the complete band gap, it is observed that radiation to the bulk of the substrate dominates. This observation is explained by introducing the concept of the sound line.

Journal Article↗

Effect of procyanidolic oligomers on corneal collagen of rabbits treated by excimer laser photoablation.

Procyanidolic oligomers (PCO) are mainly used for their therapeutic effect on the vascular wall. We could show that the mechanism of this effect involves interactions with mesenchymal cells and extracellular matrix (ECM). Recently we demonstrated in vitro that they also act on cornea, a tissue rich in ECM. For instance they stimulate the corneal biosynthesis of type VI collagen and proteoglycans. A potent antiprotease effect also could be demonstrated on corneas. In our present work we examined in vivo action of PCO on corneas. A group of rabbits received during 1 week on their right eye a treatment by procinaidolic oligomers, the left eyes were kept as controls. After 1 week both eyes underwent excimer laser photobalation. Another group of rabbits also received the PCO treatment on the right eye, not on the left, but was not treated by photoablation. One week after the surgical intervention corneas were collected and biochemical and morphological observations were carried out. Ocular administration of PCO was well tolerated and no toxic or inflammatory side-effects could be seen. In the control group photoablation was followed by a decrease of the content of corneas in type I collagen and a strong increase in type III collagen. On the corneas treated by PCO these alterations of the composition were not observed. These results indicate that PCO treatment before excimer laser photoablation maintains within normal limits the biochemical composition of the operated corneas.

Animals↗

Effect of high glucose concentration on corneal collagen biosynthesis.

The effect of high glucose concentration (3 g/l) on bovine corneal total protein and collagen biosynthesis was studied, using 3H-proline incorporation in explant cultures with protein and collagen determinations. The high glucose concentration increased the incorporation of 3H-proline in total corneal proteins as well as in collagens. The specific radioactivity of stromal collagens was strongly increased in these conditions. Mannitol was used to control the osmotic effect of the high glucose concentration, both at 1 and 3 g/l concentrations. Mannitol did not increase the incorporation of 3H-proline in total proteins or collagens, but on the contrary decreased it. The high glucose concentration decreased the excretion of neosynthesized proteins and collagens in the culture medium, but did not affect the total protein or collagen content of the corneas. The strong increase in the specific radioactivity of corneal collagens in the presence of 3 g/l glucose suggests an increased turnover of collagens in diabetic corneas. The increased biosynthesis of collagens together with their decreased elimination in the extracellular compartment can create the conditions for the formation and accumulation of advanced glycation endproducts by the Maillard reaction. This can induce and stimulate the liquefaction of the vitreous body leading to sight-threatening disorders such as diabetic retinopathy, retinal detachment, glaucoma, cataract formation and age-related macular degeneration.

Animals↗

MMP-type endopeptidase activity in the cornea. Its evolution during organ culture storage at the Eye Bank. Effect of hyaluronan.

About 46% of total corneas obtained from donors in the French Eye Bank cannot be grafted for several reasons as loss of endothelium or other. Corneal cells express proteolytic enzymes, essentially matrix metallo-proteinase MMP-2 and MMP-9. In presence of hyaluronan and some other GAG-s their activity increases as could be shown on keratocyte cultures. Hyaluronan concentration increases during in vitro preservation and can represent a serious hazard for corneal conservation. The control of MMP-release and activation might well be one of the factors involved in graft deterioration. We could show however that only a slight fraction (< or =12%) of total, relatively high endopeptidase activity of the cornea is released in the media during storing. It appears therefore that most of the proteolytic activity determined in corneal extracts remains confined to the stroma and might not represent an important risk for preservation, at least for the endothelium.

Cornea↗

Effect of procyanidolic oligomers of Vitis vinifera on the biosynthesis and excretion of corneal glycosaminoglycans.

Glycosaminoglycans (GAG) as hyaluronan and proteoglycans (PG) as dermatan sulfate (DSPG) and keratan sulfate (KSPG) play important role in the structure and physiological functions of cornea. Several corneal pathologies are associated with qualitative and quantitative alterations of GAG and/or PG biosynthesis. For these reasons we investigated the effect of procyanidolic oligomers (PCO) on the biosynthesis and excretion of GAG-s in bovine corneal explant cultures. This was accomplished by the determination of their chemical components, hexoses, hexosamines and uronic acids as well as the incorporation of (35)S-sulfate in the presence and without PCO in the culture medium. In presence of 1 mg/ml PCO total hexoses, hexosamines and uronic acids increased at 5 and 24 h of incubation, hexoses less than the two other components. Stimulation of biosynthesis concerned essentially the uronic acid containing GAG-s, DS and hyaluronan. (35)S-sulfate incorporation decreased in presence of PCO, showing a decreased sulfation of GAG-s. The most striking effect was however the strongly increased excretion of neosynthesized GAG-s in the culture medium. This effect could be beneficial in decreasing corneal hydration in inflammatory conditions, together with the previously demonstrated protection of corneal macromolecules against proteolytic degradation.

Animals↗

[Effect of procyanidolic oligomers on corneal collagen fibrillogenesis].

PURPOSE: We showed in a previous study that procyanidolic oligomers (PCOs) from grape seeds effectively protect corneal stroma against degradation by bacterial collagenase. Here we report the study of the effect of PCOs on protein and collagen composition of cornea and on the biosynthesis of corneal collagens. METHODS: Bovine corneas were used in explant cultures. We quantitatively determined total proteins and collagen as well as the incorporation of 3H-proline in separated collagen types. Collagens type I, V, and VI were separated and quantitated. In order to understand some of the results obtained, we studied the interaction of PCO with collagen type I separately. RESULTS: In the absence of PCOs, collagen typing and 3H-proline incorporation yielded the expected results for a normal cornea, with the usual proportion of the three major types of collagens. In presence of PCOs at 1 mg/ml and after 24 h incubation, total proteins and collagens decreased, as did papain-extractable collagens. Proteins in the final residue solubilized in 1 M KOH-80% v/v aqueous ethanol increased. The proportion of the three principal collagens was also modified: type I became preponderant, and the proportions of the two others (type V and VI) decreased. The study of the interaction of collagen type I with PCOs showed that 30% of total PCOs do not interact with collagen, 20% interact reversibly, and 50% of PCOs are strongly and irreversibly fixed. This strongly fixed fraction could not be separated from collagen by either column chromatography or collagenase or KOH in aqueous ethanol. CONCLUSIONS: Bovine corneas in explant cultures in presence of PCOs undergo a modification of their protein and collagen content, with a concurrent modification of the proportion of collagens types I, V, and VI. Collagen type I predominates, and the two other types decrease. Study of the collagen type I-PCO interaction showed that roughly 50% of PCOs become irreversibly fixed to collagen, resisting collagenase or other methods of separation. This strong PCO-collagen type I interaction explains the decrease in solubility.

Animals↗

Effect of a preparation containing a fucose-rich polysaccharide on periorbital wrinkles of human voluntaries.

BACKGROUND: Skin aging is accompanied by wrinkle formation. At some sites, as the periorbital skin, this is a relatively early phenomenon, variable from one woman to the other and even between the right and left eye. PURPOSE: We tested the effect of a preparation with fucose-rich oligo- and polysaccharide (FROP-3) on periorbital wrinkles (crow's feet) on a team of voluntaries. METHOD: The efficiency of a preparation containing as active principle a fucose-rich polysaccharide in a base-cream for the treatment of temporal periorbital wrinkles on 20 voluntary women was tested as follows: the women applied the cream twice a day for 4 weeks. Negative replicas were taken on the periorbital skin before and after 4 weeks of application of the cream. The results were evaluated by semi-automated morphometry on the plastic replicas and expressed as the 'wrinkle-factor' obtained by multiplying total wrinkle length in millimeters by the average wrinkle width. RESULTS: After 4 weeks of treatment, there was a significant improvement of the periorbital wrinkles for the majority of the voluntary team who tested the cream. On the 29 periocular wrinkles examined 65% showed an improvement at the end of the treatment. In two cases, the improvement was 100% on one eye (disappearance of the crow's feet) and of 75% and 79% on the other eye. On six eyes, there was no significant change after the end of the treatment, and on four eyes a worsening was observed. The age-dependent modifications showed that improvement was independent of age and was the result of individual skin reactions to the cream. The occurrence and the severity of crow's feet is highly individual, and differs from one side of the face to the other for the same person. These strong individual variations explain the highly variable results, not only from one woman to the other but also comparing one side of the face to the other for the same person. CONCLUSION: Under the effect of 4 weeks of treatment with the FROP-3-containing cream, most of the voluntaries had their periorbital wrinkles attenuated and some women showed a total regression of crow's feet on one of their eyes.

Adult↗

Effects of L-fucose and fucose-rich oligo- and polysaccharides (FROP-s) on collagen biosynthesis by human skin fibroblasts. Modulation of the effect of retinol, ascorbate and alpha-tocopherol.

The action of L-fucose and a fucose-rich oligosaccharide (FROP-3) on skin explant cultures and fibroblast cell cultures, alone or together with three vitamins (A, C, E, often used in topical preparations) was studied, using as criteria collagen accumulation and collagen and non-collagen protein biosynthesis. In order to confirm the results observed, several variants of the chemical procedure were used. Collagen accumulation in cell cultures and (3)H-proline incorporation in proteins and collagen gave comparable results. L-Fucose produced a slight but significant inhibition of collagen accumulation and of incorporation of radioactive proline in cell associated collagen and also modified the ratio of excreted (soluble) to cell- or tissue-bound collagen and non-collagen proteins. The oligosaccharide (FROP-3) did not produce a comparable inhibition of collagen biosynthesis. Both L-fucose and FROP-3 modulated the action of the two above-mentioned vitamins in most experimental conditions used. The combined action of the three vitamins (all-trans retinol, ascorbate, alpha-tocopherol) with L-fucose and even more so with FROP-3 can be considered as favourable for the modulation of the biosynthetic activity of fibroblasts.

Ascorbic Acid↗

Pharmacology of skin aging. Stimulation of glycosaminoglycan biosynthesis by L-fucose and fucose-rich polysaccharides, effect of in vitro aging of fibroblasts.

The effect of L-fucose and fucose-rich polysaccharides (FROP-s [Biomed. Pharmacother., 2003; 57: 187-94]) was investigated, using human skin fibroblast cultures at several passages. The cells were incubated with radioactive glucosamine for 24 h, followed by the determination of individual glycosaminoglycans (GAG-s) by selective hydrolysis using specific enzymes. The effect of L-fucose and of FROP-3 [Biomed. Pharmacother., in 2003; 57: 187-94], both at 1 and 10 microg/ml, added to the culture medium, was investigated. L-Fucose stimulated the incorporation of the tracer in heparan sulfates by fibroblasts at the ninth passage by 20%. FROP-3 stimulated incorporation in keratan sulfates by 45% by fibroblasts of the fifth passage. This effect was identical at both concentrations tested. For fibroblasts at the ninth passage FROP-3 stimulated incorporation in dermatan sulfate. This effect was dose dependent of the order of +67% at 1 microg/ml and +128% at 10 microg/ml. Incorporation in hyaluronan was also stimulated by about +27%. These stimulations of GAG-biosynthesis might play a role in the increase of total skin thickness of hairless rats treated with L-fucose, as well as in several other favorable results recorded for FROP-3 such as the increased hydration (resistance to pressure) and elasticity of human skin (Robert C, Robert AM, Robert L).

Cells, Cultured↗

Effect of L-fucose and fucose-rich polysaccharides on elastin biosynthesis, in vivo and in vitro.

With increasing age elastic fibres in human skin are progressively lysed and skin elasticity is also decreasing. Still there is an age-dependent increase of elastic fibre surface density, mostly due to an alteration of the fibres. The present experiments were undertaken to explore if L-fucose and fucose-rich polysaccharides (FROP-s) could influence elastin biosynthesis. We show here, that topical application of a fucose-containing preparation to the skin of hairless rats increased after 4 weeks the elastic fibre surface density by about 40%, shown by quantitative morphology. Using human skin fibroblasts in explant cultures, the addition of L-fucose or of FROP-3 increased the biosynthesis of immunoprecipitable tropoelastin by about 40%. No increase was found however of desmosine-isodesmosine in skin explant cultures after 72 h of incubation. The effect of L-fucose and FROP-3 on the biosynthesis of collagen and non-collagen proteins excreted by the skin explant cultures was also investigated. L-fucose, but not FROP-3, decreased collagen biosynthesis but both increased non-collagen protein biosynthesis. These results show that L-fucose and FROP-3 stimulate tropoelastin biosynthesis in vitro, and elastic fibre formation in vivo. This stimulation concerns also several non-collagen proteins secreted by skin explant cultures. Elastic fibre formation necessitates the simultaneous synthesis of several microfibrillar glycoproteins as well as of tropoelastin. The increased elastic fibre density in the in vivo experiments suggests that this is indeed achieved by L-fucose and FROP-3, further demonstrating their efficiency in the control of age-dependent modifications of connective tissues in general and of skin in particular.

Administration, Cutaneous↗

Combination of raltitrexed and oxaliplatin is an active regimen in malignant mesothelioma: results of a phase II study.

PURPOSE: The aim of this open-label phase II study was to evaluate the activity of raltitrexed (Tomudex; AstraZeneca, Cergy, France) and oxaliplatin combination therapy in patients with diffuse malignant pleural mesothelioma. PATIENT AND METHODSs: Fifteen pretreated and 55 chemotherapy-naive patients (median age, 60 years; World Health Organization performance status of < or = 2) were enrolled. Most patients (66%) had advanced disease. Patients received raltitrexed 3 mg/m2 followed by oxaliplatin 130 mg/m2 every 3 weeks. RESULTS: Twenty-four patients (34%) were classified as having a poor prognosis. In the overall study population, 14 patients (20%) had a partial response, and 32 patients (46%) had stable disease. The symptomatic response rates were as follows: shortness of breath, 36%; pain, 30%; activity, 23%; appetite, 21%; and asthenia, 20%. Median time to disease progression was 18 weeks (95% confidence interval [CI], 13 to 22 weeks). In chemotherapy-naive patients, median survival was 31 weeks (95% CI, 23 to 40 weeks) from the start of treatment and 49 weeks (95% CI, 40 to 52 weeks) from diagnosis of mesothelioma. In pretreated patients, median survival was 44 weeks (95% CI, 24 to 40 weeks) from the start of treatment and 226 weeks (95% CI, 63 to 292 weeks) from the diagnosis of mesothelioma. Overall 1-year survival was 26% (95% CI, 15.5% to 36.4%), survival was 22% (95% CI, 10.9% to 33.2%) in chemotherapy-naive patients and 40% (95% CI, 15.2% to 64.8%) in pretreated patients. Hematologic toxicity was mild, and there was no alopecia. The most common adverse events were asthenia, nausea/vomiting, and paraesthesia, and no treatment-related deaths were reported. CONCLUSION: The raltitrexed and oxaliplatin combination is an active outpatient regimen in malignant mesothelioma and has an acceptable tolerability profile.

Adult↗