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Biomedical subjects

G Goffinet

Publications and source records attributed to G Goffinet.

At least 19 recordsLinked to original sources
PubMed2003

Characterization of the primary sonic muscles in Carapus acus (Carapidae): a multidisciplinary approach.

Sound production in carapid fishes results from the action of extrinsic muscles that insert into the swim bladder. Biochemical, histochemical and morphological techniques were used to examine the sonic muscles and compare them with epaxial muscles in Carapus acus. Sonic fibres are thicker than red and thinner than white epaxial fibres, and sonic fibres and myofibrils exhibit an unusual helicoidal organization: the myofibrils of the centre are in a straight line whereas they are more and more twisted towards the periphery. Sonic muscles have both features of red (numerous mitochondria, high glycogen content) and white (alkali-stable ATPase) fibres. They differ also in the isoforms of the light chain (LC3) and heavy chain (HC), in having T tubules at both the Z-line and the A-I junction and in a unique parvalbumin isoform (PAI) that may aid relaxation. All these features lead to the expression of two assumptions about sound generation: the sonic muscle should be able to perform fast and powerful contractions that provoke the forward movement of the forepart of the swim bladder and the stretching and "flapping" of the swim bladder fenestra; the helicoidal organization allows progressive drawing of the swim bladder fenestra which emits a sound when rapidly released in a spring-like manner.

Air Sacs↗
PubMed2003

Effect of ophthalmic viscosurgical devices on lens epithelial cells: a morphological study.

PURPOSE: To investigate the morphological effects of Viscoat (sodium hyaluronate 3.0%-chondroitin sulfate 4.0%) on lens epithelial cells (LECs). SETTING: Department of Ophthalmology, University of Texas Southwestern Medical Center, Dallas, Texas, USA, and the Laboratory of Ultrastructural Morphology, Zoological Institute, University of Liège, Liège, Belgium. METHODS: Human LECs collected via capsulorhexis were examined by light microscopy (LM) and transmission electron microscopy (TEM). Lens epithelial cells from rabbit capsulorhexis samples were studied by LM and TEM following exposure to Provisc (sodium hyaluronate 1.0%) or Viscoat ophthalmic viscoelastic device (OVD). Since Viscoat is hypertonic (340 mOsm), hypertonic, isotonic, and hypotonic solutions were compared to investigate a possible mechanism for the observed effects. The effects of Provisc and Viscoat on rabbit LECs in the intact lens were also compared. RESULTS: Human LECs gathered via capsulorhexis following exposure to Viscoat were generally thinner than control samples and often had condensed nuclei and increased intracellular vacuolization. Rabbit capsular tissue exposed in situ to Viscoat demonstrated changes similar to those seen in humans. Cells exposed to Provisc were similar to cells in untreated controls in humans and rabbits. Corneal endothelial cells exposed to either agent were unaffected. Experiments with hypertonic and hypotonic buffers induced some of the changes noted with Viscoat, but the effects were less severe. Lens epithelial cells in intact rabbit lenses exposed to Viscoat appeared similar to LECs in the control samples. CONCLUSIONS: Light microscopy and TEM of human lens capsule tissue suggest that Viscoat induces significant morphological changes in LECs during cataract surgery. The changes may underlie the improved visualization of these cells that has been reported during cataract surgery. Corneal endothelial cells were unaffected by exposure to Viscoat. Studies in a rabbit model suggest that the hyperosmolarity of Viscoat may play a partial role in the LEC changes.

Animals↗
PubMed2001

Preparation of poly(D,L-lactide) nanoparticles assisted by amphiphilic poly(methyl methacrylate-co-methacrylic acid) copolymers.

When co-precipitated with amphiphilic copolymers from DMSO, poly(D,L-lactide) (PLA) can be readily converted into stable sub-200 nm nanoparticles by addition of an aqueous phase, free of any polymeric stabilizers such as poly(vinyl alcohol) or Poloxamer. In this work, the ability of random poly(methyl methacrylate-co-methacrylic acid) copolymers (PMMA-co-MA) to stabilize PLA nanoparticles was demonstrated, and the properties of PLA/PMMA-co-MA nanoparticles were investigated. When co-precipitated with PMMA-co-MA, PLA was totally converted into nanoparticles using a polymer concentration in DMSO (Cp) below 17.6 mg ml(-1), and a PMMA-co-MA proportion above a critical value depending on the content of MA repeating units (X). For instance, the lowest PMMA-co-MA proportion required was 0.9 mg mg(-1) PLA for X = 12%, and 0.5 mg mg(-1) PLA for X = 25% (for C(PLA) = 16 mg ml(-1) DMSO). The nanoparticle diameter was essentially independent of X, the proportion of PMMA-co-MA, and the PLA molecular weight, except for oligomers for which the nanoparticle diameter was smaller. It decreased when the organic phase was diluted (126 +/- 13 nm for Cp = 17.6 mg ml(-1), and 81 +/- 5 nm for C(P) = 5.6 mg ml(-1)). The time-dependence of the stability and the degradation of PLA/PMMA-co-MA nanoparticles was discussed. One of the main advantages of this technique is the ability to control surface properties and to bring functional groups to otherwise non-functionalized PLA nanoparticles. To illustrate this, a conjugate of PMMA-co-MA25 and biotin was synthesized, and used to prepare biotinylated nanoparticles that could be detected by fluorescence and transmission electron microscopy after infiltration into ligatured rat small intestine.

Animals↗
PubMed1999

Immunocytochemistry of lipids: chemical fixatives have dramatic effects on the preservation of tissue lipids.

We report here the effects of chemical fixatives on lipids studied under conditions simulating the immunogold labelling of phosphatidylserine. Using anti-phosphatidylserine antibodies, it is shown that the labelling intensity of a phosphatidylserine/phosphatidylcholine coating depends largely on the conditions of fixation. In fact, the usual aldehydic fixatives washed out most of the phostphatidylserine, thus preventing the binding of anti-phosphatidylserine antibodies. This was confirmed on biological samples such as rat liver and brain by measuring the loss of radiolabelled lipids during the fixation procedure. Furthermore, the complete procedure of tissue preparation for electron microscopical observation was investigated. The loss of (radiolabelled) lipids was studied in tissue samples during fixation and resin embedding. The results showed that the classical procedure (glutaraldehyde fixation followed by epoxy resin embedding) results in the loss of 73-91% of the tissue lipids whereas in unfixed, freeze-substituted samples, more than 76% of the tissue lipids are preserved.

Animals↗
PubMed1999

In vitro stimulation of human gingival epithelial cell attachment to dentin by surface conditioning.

BACKGROUND: Chemical root conditioning is widely used to improve the outcome of regenerative periodontal therapies by favoring the attachment of the regenerated periodontal structures. Although the effect of root conditioning on periodontal mesenchymal cells is well documented, very little is known about its potential effect on the re-formation of the junctional epithelium, a crucial event for the protection of the wound. The goal of the present study was to test in vitro the consequences of dentin conditioning with citric acid or minocycline on the attachment kinetics and morphology of human gingival keratinocytes (HGK). METHODS: The attachment kinetics of HGK to samples of powdered human dentin (particle size 44 to 76 microm) were examined by use of 3H-labeled cells. The morphology of attached epithelial cells was then determined by scanning electron microscopy (SEM). RESULTS: When the initial adhesion kinetics of cells on untreated dentin were tested, the percentage of attached HGK proved to be dependent on the number of plated cells and the time of incubation (from 0 to 12 hours). Conditioning the dentin by 3% citric acid or by minocycline-HCl (at 0.01, 0.1, or 2.5%) significantly increased (P <0.005) keratinocyte attachment beyond 6 hours, without notable differences between the 2 substances at any concentration. The attachment kinetics of HGK preincubated for 24 hours by 10 microg/ml minocyline-HCl on untreated dentin was found to be similar to that observed for non-preincubated cells. These results are in agreement with the SEM observations: indeed, the surface conditioning of dentin significantly modified the morphology of attached HGK, whereas the preincubation of these cells with minocyline-HCl did not. CONCLUSIONS: These results suggest that minocycline-HCl does not exert a direct effect on human gingival epithelial cells. In contrast, conditioning the dentin by citric acid or by minocycline stimulates the attachment of HGK, which could lead to a rapid periodontal healing by favoring the re-formation of a junctional epithelium.

Anti-Bacterial Agents↗
PubMed1998

Comparison between the natural postnatal maturation and the spermine-induced maturation of the rat intestine.

In the suckling rats, orally provided spermine induced structural and biochemical changes in the intestine, which are characteristics of the postnatal maturation. This induced maturation was compared to that occurring spontaneously. Eight mumol spermine were administered orally once a day, for one or three days, to suckling rats which were 11 days old at the beginning of the experiment. The animals were killed 0, 2, 4, 6, 8, 10 hours or 3 days after the first treatment. Control rats from the same litter were treated in the same way but received only the vehicle. In order to complete the study of the naturally occurring maturation, another group of rats was killed when they were 12, 13, 15, 16, 17, 18, 19, 20, 21 or 30 days old. Animal and intestine weights were measured. Disaccharidase specific activity, and protein, DNA and RNA contents were estimated in the small intestine. Histological and ultrastructural aspects of the intestinal mucosa were examined. For all these parameters, the maturation induced by spermine ingestion appeared close to that occurring naturally at weaning. Consequently, dietary spermine induces all the morphological and biochemical modifications characterizing the intestinal postnatal maturation in the suckling rat suggesting a role of the polyamines in the naturally occurring processes.

Animals↗
PubMed1998

In vitro modulation of human gingival epithelial cell attachment and migration by minocycline-HCL.

Although the influence of tetracyclines on periodontal connective tissue cells has been the topic of many in vitro and in vivo studies, data regarding their effects on gingival epithelial cells are scarce. The present in vitro study was designed to examine the influence of minocycline, a semi-synthetic analog of tetracycline, on human gingival keratinocyte (HGK) attachment and migration. Attachment tests were performed with HGK prelabeled by tritiated amino-acids. Increasing concentrations of minocycline (10, 50, 100 micrograms/ml) in the medium produced no significant modification of cell adhesion kinetics compared to control conditions, except for 100 micrograms/ml which statistically significantly (p < 0.05) reduced the number of attached cells beyond 6 h. A 24-h cell preincubation in 10 micrograms/ml of minocycline did not alter the kinetics of HGK attachment. Scanning electron microscopic observations of attached HGK showed that the presence of 10 micrograms/ml of minocycline in the "attachment medium" induced the production of multiple filopodial extensions. Migration tests in Boyden chambers for 40 h demonstrated that HGK preincubation for 24 h in a 10 micrograms/ml minocycline-HCl solution increased significantly (p < 0.005) cell migration towards a gradient of fetal calf serum. The presence of 10 micrograms/ml of minocycline in contact with the keratinocytes in the upper compartment of the migration chambers also produced a significant (p < 0.005) result. In contrast, the presence of minocycline in the lower compartments did not produce any chemoattractive effect. Within the limits of their significance, these results suggest that, at concentrations not beyond 50 micrograms/ml, minocycline could fasten the periodontal wound coverage by epithelial cells and allow the normal reformation of a junctional epithelium.

Adolescent↗
PubMed1997

"Exolysosomes," enzyme-containing vesicles in the ecdysial space of molting crabs.

Free vesicle-like bodies (VLBs) present in the ecdysial space of cuticle regions undergoing degradation during preecdysis of the Atlantic shore crab Carcinus maenas have been interpreted either as infectious organisms or as secretion structures associated with degradation of the old cuticle. Ultrastructural, cytochemical, and immunocytological investigations were performed to test these hypotheses and to see whether VLBs are peculiar to this crab species. Similar VLBs were systematically found in two other preecdysial crabs, Cancer pagurus and Macropipus puber. In Car, maenas, they originate during early premolt inside Golgi buddings and are often gathered into large vacuoles in epidermal cells. The histochemical azo-dye technique and a cerium-based cytochemical method revealed acid phosphatase activity in both the ecdysial space and the VLBs, while Feulgen's method and immunocytological labeling always failed to reveal any DNA or RNA in either the ecdysial space or the VLBs. We conclude that VLBs are not infectious organisms but "extracellular" cuticle-degrading organelles of lysosomal origin and propose to coin them "exolysosomes."

Acid Phosphatase↗
PubMed1997

Scintigraphic visualization of intrathecal liposome biodistribution.

BACKGROUND: Liposomes containing local anaesthetics have been administered intrathecally and in the epidural space. Poor attention has been given to the pharmacokinetics of liposomes as drug carriers. Therefore, we observed the biodistribution of liposomes after intrathecal injection in rats by scintigraphic imaging during 24 h. METHODS: We administered 99mTc-labeled multilamellar (MLV) and small unilamellar vesicles (SUV) of defined size and volume dispersities into the cerebrospinal fluid at the lumbar level. Those vesicles were free of contamination by radiolabeled colloids as visualized by light and electron microscopy and of neurotoxic products from phosphatidylcholine hydrolysis and peroxidation, both during the preparation process and after 24 h incubation in cerebrospinal fluid at 37 degrees C in vitro. RESULTS: SUV immediately diffused from the lumbar site of injection to the head and were cleared between 1 and 24 h after injection. MLV were cleared more slowly from the spinal space and appeared in the head region 1 h after injection where they accumulated up to 24 h. These differences were explained in terms of vesicle sizes and volumes. SUV with 0.05 micron diameters were rapidly absorbed into the blood through the arachnoid granulations. In contrast, particles larger than the upper size limit of the arachnoid granulations permeability (+/- 8 microns) could accumulate in the head with a slow elimination rate. CONCLUSION: This difference in clearance from the intrathecal space outlines the importance of defining the size of the liposomes, the distribution of a tracer or a drug inside the liposomal preparation, the chemical stability and the absence of toxic degradation products of liposome formulations before clinical use.

Animals↗
PubMed1996

Ultrastructural modifications in cultured fetal quail hepatocytes exposed to pesticides and PCBs.

There is increasing interest in cultured hepatocytes as a tool for solving toxicological and pharmacological problems while reducing laboratory animal experimentation. In the present study, fetal hepatocytes from the Japanese quail (Coturnix coturnix japonica) were used as an in vitro alternative model for evaluating the effects of PCBs and various pesticide-type chemicals on cell ultrastructure. Major alterations were demonstrated. The most striking effects of toxicants were an increase in the number of cisternae of the rough endoplasmic reticulum (RER), various alterations of mitochondrial morphology, a decreased glycogen content, vacuolization of the cytoplasm, and the appearance of concentric membrane arrays (CMA's), also called myelin-like figures. Other changes were sometimes observed, such as altered cell junctions, an increased lipid content, deformations of the nuclei, or the appearance of crystalline structures. These ultrastructural modifications seem to be dose-dependent. The present in vitro findings are validated by similar observations previously made in vivo on Japanese quail. They confirm the effectiveness of this technique as a biomonitoring tool for the evaluation of environmental quality. Yet the multiplicity of possible toxic effects, even for xenobiotics of a same category, makes it necessary to screen additional indicators of toxicity, such as the detoxifying activity of monooxygenases.

Animals↗
PubMed1996

99mTechnetium-stannous oxinate as marker of liposome formulations.

Liposomes associated with tin(II) dioxinate were prepared from egg yolk phosphatidylcholine and cholesterol as sterile and pyrogen-free multilamellar or unilamellar vesicles. Complexing of liposomal tin(II) dioxinate with 99mTc attained 98% of the added radioactivity. Thirty percent 99mTc were released during 24-h incubation in biological fluids. The absence of tin colloids seen by electron microscopy and the stability of liposomal phospholipid and tin(II) dioxinate during 72-h incubation at 37 degrees C in plasma and cerebrospinal fluid would allow safe and reliable scintigraphic liposome pharmacokinetic studies.

Cholesterol↗
PubMed1995

Immunogold labelling of fatty acyl chains.

For the first time, antibodies against a hydrophobic hapten have been used for immunogold labelling of a lipid antigen (BSA-C18:1 conjugate) coated on polystyrene. The labelling was visualised either directly in transmission electron microscopy or in light microscopy after silver enhancement. Good recognition of the fatty acyl chain was obtained even after treatment of the antigen coat with various cross-linking fixatives used for electron microscopy, i.e. formaldehyde, glutaraldehyde and osmium tetroxide.

Fatty Acids↗
PubMed1995

Cytotoxic effects of Aroclor 1254 on ultrastructure and biochemical parameters in cultured foetal rat hepatocytes.

The cytotoxicity of a commercial PCB mixture, Aroclor 1254, was assessed on cultured foetal rat hepatocytes. Under control conditions, dexamethasone stimulates immature hepatocytes to differentiate into both hepatocytes and biliary epithelial cells. Consequently, foetal rat hepatocytes maintain, in vitro, a liver-like organization with spaces corresponding to the lumen of biliary canalicules, many mitochondria, and a well-developed rough endoplasmic reticulum (RER). This in vivo-like organization of cultured rat hepatocytes remains unchanged in medium supplemented with Aroclor 1254 at concentrations below 25 microM. In the 25-125 microM concentration range, however, PCBs severely alter some cellular organelles, notably causing important development of the RER and the appearance of cytoplasmic lacunae containing laminated concentric membrane arrays. In addition, the number of lipid droplets increases, the glycogen islets disappear, and dramatic local alterations of the mitochondrial cristae occur. In exposed and unexposed cells, the following biochemical parameters were measured: the DNA content, protein synthesis, lipid peroxidation, and urea formation. The results show that Aroclor 1254 at concentrations exceeding 25 microM (but not at lower concentrations) causes irreversible damage to cultured hepatocytes. The observed ultrastructural modifications are in good agreement with several in vivo studies on rat liver. Thus, isolated foetal rat hepatocytes have considerable potential as an alternative to whole animals for use in (eco)toxicological studies.

Analysis of Variance↗
PubMed1995

Biodistribution of liposomes after extradural administration in rodents.

We have mapped over 24 h the biodistribution of 99mTc-labelled multilamellar and small unilamellar liposomes in rabbits and rats by scintigraphic imaging after extradural injection. Multilamellar vesicles formed a depot at the site of injection; small unilamellar vesicles spread immediately along the extradural space and entered the systemic compartment 30 min after injection. Well-delineated liver and kidney labellings were seen after 24 h. The use of 3H-cholesterol-labelled small unilamellar vesicles suggested hepatic capture of intact liposomes with sizes averaging 0.05 microns drained unmodified into the systemic circulation through the extradural lymphatics. These results have led to the selection of multilamellar vesicles (0.1-15 microns size range) for clinical trials using liposome-associated local anaesthetics.

Animals↗
PubMed1995

Biodistribution of liposome-associated bupivacaine after extradural administration to rabbits.

After one extradural injection of 0.25% bupivacaine 0.3 ml and 3H-bupivacaine 0.005 mCi in multilamellar liposomes, no systemic radioactivity (plasma, liver, heart muscle) was obtained for 1 h, and the labelling was less than that of systemic distribution of plain bupivacaine for the following 3 h. In contrast, radioactivity in the lumbar spinal nerves peaked in the first hour and remained higher than that of plain bupivacaine for 4 h. No radioactivity was measured in cerebrospinal fluid. Small unilamellar vesicles incorporating 3H-cholesterol did not significantly label spinal nerves and central nervous structures indicating that the mode of action of liposomal bupivacaine did not involve uptake by nerve structures. Rapid uptake of radioactivity by spinal nerves suggested exchange of bupivacaine between liposomes and nerve sheaths.

Anesthetics, Local↗
PubMed1994

Expression and induction of drug-metabolizing enzymes in cultured fetal rat hepatocytes.

An in vitro experimental model, fetal rat hepatocytes in culture, was metabolically characterized. Several enzymatic activities were expressed in these hepatocytes, namely, testosterone hydroxylations. Hepatocytes cultured up to 3 weeks in the presence of dexamethasone and phenobarbital still expressed some drug-metabolizing enzyme activities (e.g., ECOD). The enzymatic activities were measured both directly on monolayers during culture and on the corresponding harvested and homogenized cells. The results correlate perfectly with each other. The 'on cell' procedure allows us to repeat the assay or to measure several activities on the same cells at different time intervals. The presence of dexamethasone in the culture medium allows the expression and the induction of several cytochrome P450 isoenzymes, namely, those hydroxylating testosterone. This makes the model particularly attractive for induction experiments as well as for metabolic or toxicological studies needing longer treatments.

7-Alkoxycoumarin O-Dealkylase↗
PubMed1993

Cytokeratin in lens epithelial cells and its effect on anterior lens capsule opacification.

Postoperative lens capsule opacification is a complication of extracapsular cataract extraction. To avoid this problem, we aspirated the epithelial cells lining the internal surface of the capsular bag. Anterior capsule specimens collected three and six years after the first surgical intervention (extracapsular cataract extraction with intercapsular implantation of a poly[methyl methacrylate] intraocular lens) and in vitro studies of epithelial cell cultures revealed the presence of intracellular and extracellular cytokeratin. After several years, the cytokeratin layers were arranged in a pattern analogous to that of liquid crystalline phases and many nonhuman extraocular tissues.

Cataract↗