Changements de l'epithélium médian des bourgeons palatins de souris au stade de préfusion

Résumé Avant d'entrer en contact, les surfaces de fusion des processus palatins subissent des modifications. Certaines cellules épithéliales superficielles deviennent turgescentes, alors que d'autres conservent un aspect dense et aplati, elles se lysent et éclatent. Des noyaux semblables à ceux des assises profondes se retrouvent en surface, paraissant migrer vers le bourgeon antagoniste. Des projections filamenteuses apparaissent, quelques unes établissant un contact avec l'épithélium opposé, d'autres restant seulement sous forme d'expansion. Ces observations sont concordantes en microscopie photonique et en microscopie électronique à balayage.

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Modification of the medial epithelium of the palatal shelves of mice at the prefusion stage. A light and scanning electron microscopy study

Summary The fusion surfaces of the palatal processes are modified before coming into contact. Several superficial epithelial cells become hypertrophied, undergo lysis and burst, whereas others remain flattened and densely stained. Nuclei, similar to those localized in deeper layers are found on the surface and seem to migrate towards the opposite process. Filamentous projections appear and some establish a linkage with the opposite epithelium, whereas others expand freely from the surface. Light microscope observations agree with scanning electron microscopy findings.

     

A bone preservation protocol that enables evaluation of osseointegration of implants with micro- and nanotextured surfaces

Development of surface treatments has enabled secure attachment of dental implants in less than 1 month. Consequently, it is necessary to characterize accurately the osseointegration of the implant surface in the region of the bone-implant contact (BIC). We developed a method for sample preparation that preserves both bone and BIC to permit analysis of the contact interface. We prepared eight nanotextured implants and implanted them in rabbit tibias. After healing for 30 days, outcomes were analyzed using both our bone preservation protocol and routine decalcification followed by preparation of histological sections stained by hematoxylin and eosin (H & E). Pull-out tests for implant osseointegration were performed after healing. Non-implanted samples of rabbit mandible were used as a control for assessing organic and mineralized bone characteristics and bone structure. Our bone preservation protocol enabled evaluation of many of the same bone characteristics as histological sections stained with H & E. Our protocol enables analysis of implant samples, implant surfaces and osseointegration without risk of BIC damage.     

Photocatalytic antimicrobial activity of thin surface films of TiO(2), CuO and TiO (2)/CuO dual layers on Escherichia coli and bacteriophage T4

TiO₂-coated surfaces are increasingly studied for their ability to inactivate microorganisms. The activity of glass coated with thin films of TiO₂, CuO and hybrid CuO/TiO₂ prepared by atmospheric Chemical Vapour Deposition (Ap-CVD) and TiO₂ prepared by a sol–gel process was investigated using the inactivation of bacteriophage T4 as a model for inactivation of viruses. The chemical oxidising activity was also determined by measuring stearic acid oxidation. The results showed that the rate of inactivation of bacteriophage T4 increased with increasing chemical oxidising activity with the maximum rate obtained on highly active sol–gel preparations. However, these were delicate and easily damaged unlike the Ap-CVD coatings. Inactivation rates were highest on CuO and CuO/TiO₂ which had the lowest chemical oxidising activities. The inactivation of T4 was higher than that of Escherichia coli on low activity surfaces. The combination of photocatalysis and toxicity of copper acted synergistically to inactivate bacteriophage T4 and retained some self-cleaning activity. The presence of phosphate ions slowed inactivation but NaCl had no effect. The results show that TiO₂/CuO coated surfaces are highly antiviral and may have applications in the food and healthcare industries.     

Direct visualization of actin nematic network formation and dynamics

Various actin assemblies within the cell regulate many cellular processes such as cell shape and motility. The mechanical properties of these networks are challenging to measure in vivo. They have been studied in solution by indirect observation methods, such as multiple ball tracking. However, little is known about the behavior of such networks near the crowded cell membrane. Here we used in vitro TIRF microscopy to directly probe the formation of actin networks in real-time near a hydrophilic surface in the presence of crowding agents. We find that under these conditions actin does not form a mesh like network, but either textured nematic liquid crystals or a bundled network. We are directly able to follow the thermal fluctuations of actin filaments within these networks. Prearranged parallel networks of actin filaments near the crowded cell membrane could play a role in the rapid formation of stress fibers or microvilli.     

On the Adsorption of Human Acidic Proline-rich Proteins (PRP-1 and PRP-3) and Statherin at Solid/liquid Interfaces

The objective of the present study was to investigate the adsorption of PRP-1, PRP-3 and statherin to solid surfaces in terms of dependence on concentration, the presence of electrolyte and surface wettability. Time resolved in situ ellipsometry was used to determine the adsorbed amounts and adsorption rates of pure PRP-1, PRP-3 and statherin onto pure (hydrophilic) and methylated (hydrophobized) silica surfaces. The initial film build-up was fast and plateaus were reached within 10 min at all concentrations for both types of surfaces and all proteins. The observed adsorption and calculated diffusion rates of PRP-1, PRP-3 and statherin, respectively, indicated that the initial adsorption was mass transport controlled at low concentrations. At hydrophobic surfaces, isotherm shapes and adsorbed amounts were similar for PRP-1 and PRP-3, while statherin adsorbed to a higher extent. At hydrophilic surfaces only PRP-1 adsorbed substantially, while for PRP-3 and statherin adsorbed amounts were low. The presence of Ca 2+ ions in the phosphate buffer solution increased the adsorption of statherin and PRP-3 on hydrophobic surfaces, while PRP-1 was unaffected. On hydrophilic surfaces, all three proteins adsorbed in higher amounts in NaCl, compared to CaCl 2 at similar ionic strength. It is concluded that acidic PRPs (PRP-1 and PRP-3) and statherin readily form films on a variety of materials and solution conditions, showing that their functions may be fulfilled under a wide range of conditions.     

Killing bacteria present on surfaces in films or in droplets using microwave UV lamps

The killing of bacteria on surfaces by two types of UV sources generated by microwave radiation is described. In both cases, UV radiation is produced by gas-discharge electrodeless lamps (Ar/Hg) excited by microwaves generated by a power supply from a standard domestic microwave oven. For UV lamp excitation, one of these sources makes use of a coaxial line with a truncated outer electrode that allows the excitation of gases and gaseous mixtures over a wide range of pressures at a comparatively low microwave power. In the second source, UV lamps are placed inside a microwave oven. Ultraviolet generated by the two sources was used to destroy vegetative Escherichia coli bacteria dispersed in thin films and in droplets on surfaces. Two types of UV lamps were used in the study. The first was constructed of quartz that filtered UV below 200 nm preventing the dissociation of oxygen in air and, hence, ozone production. The second type of tube was transparent to UV below 200 nm facilitating ozone production in air surrounding it. It was shown that bacterial cells dispersed in films on surfaces are killed more rapidly than cells present in droplets when using the lamps producing ozone and UV radiation. The UV sources described can effect rapid killing and constitute a cost-effective treatment of food and other surfaces, and, the destruction of airborne viruses and bacteria. The lamps can also be utilised for the rapid eradication of microorganisms in liquids.     

Engineering Microtools in Polymers to Study Cell Biology

Recent advances in surface engineering and soft lithography provide tools to fabricate patterned surfaces and microfluidic devices with dimensions comparable to the sizes of single mammalian cells. These technologies enable the studies of individual cells on spatially well‐defined, patterned surfaces, and in contact with patterned liquid media. They provide information about cells impossible to obtain from traditional biochemical techniques.     

An Activated GOPS‐poly‐L‐Lysine‐ Coated Glass Surface for the Immobilization of 60mer Oligonucleotides

To explore a method for enhancing the immobilization and hybridization efficiency of oligonucleotides on DNA microarrays, conventional protocols of poly‐L‐lysine coating were modified by means of surface chemistry, namely, the slides were prepared by the covalently coupling of poly‐L‐lysine to a glycidoxy‐modified glass surface. The modified slides were then used to print microarrays for the detection of the SARS coronavirus by means of 60mer oligonucleotide probes. The characteristics of the modified slides concerning immobilization efficiency, hybridization dynamics, and probe stripping cycles were determined. The improved surface exhibited high immobilization efficiency, a good quality uniformity, and satisfactory hybridization dynamics. The spotting concentration of 10 μmol/L can meet the requirements of detection; the spots were approximately 170 nm in diameter; the mean fluorescence intensity of the SARS spots were between 3.2 × 10⁴ and 5.0 × 10⁴ after hybridization. Furthermore, the microarrays prepared by this method demonstrated more resistance to consecutive probe stripping cycles. The activated GOPS‐PLL slide could undergo hybridization stripping cycles for at least three cycles, and the highest loss in fluorescence intensity was found to be only 11.9 % after the third hybridization. The modified slides using the above‐mentioned method were superior to those slides treated with conventional approaches, which theoretically agrees with the fact that modification by surface chemistry attaches the DNA covalently firmly to the slides. This protocol may have great promise in the future for application in large‐scale manufacture.     

Statistical analysis of interface similarity in crystals of homologous proteins

Many proteins function as homo-oligomers and are regulated via their oligomeric state. For some proteins, the stoichiometry of homo-oligomeric states under various conditions has been studied using gel filtration or analytical ultracentrifugation experiments. The interfaces involved in these assemblies may be identified using cross-linking and mass spectrometry, solution-state NMR, and other experiments. However, for most proteins, the actual interfaces that are involved in oligomerization are inferred from X-ray crystallographic structures using assumptions about interface surface areas and physical properties. Examination of interfaces across different Protein Data Bank (PDB) entries in a protein family reveals several important features. First, similarities in space group, asymmetric unit size, and cell dimensions and angles (within 1%) do not guarantee that two crystals are actually the same crystal form, containing similar relative orientations and interactions within the crystal. Conversely, two crystals in different space groups may be quite similar in terms of all the interfaces within each crystal. Second, NMR structures and an existing benchmark of PDB crystallographic entries consisting of 126 dimers as well as larger structures and 132 monomers were used to determine whether the existence or lack of common interfaces across multiple crystal forms can be used to predict whether a protein is an oligomer or not. Monomeric proteins tend to have common interfaces across only a minority of crystal forms, whereas higher-order structures exhibit common interfaces across a majority of available crystal forms. The data can be used to estimate the probability that an interface is biological if two or more crystal forms are available. Finally, the Protein Interfaces, Surfaces, and Assemblies (PISA) database available from the European Bioinformatics Institute is more consistent in identifying interfaces observed in many crystal forms compared with the PDB and the European Bioinformatics Institute's Protein Quaternary Server (PQS). The PDB, in particular, is missing highly likely biological interfaces in its biological unit files for about 10% of PDB entries.     

Inference of macromolecular assemblies from crystalline state

We discuss basic physical-chemical principles underlying the formation of stable macromolecular complexes, which in many cases are likely to be the biological units performing a certain physiological function. We also consider available theoretical approaches to the calculation of macromolecular affinity and entropy of complexation. The latter is shown to play an important role and make a major effect on complex size and symmetry. We develop a new method, based on chemical thermodynamics, for automatic detection of macromolecular assemblies in the Protein Data Bank (PDB) entries that are the results of X-ray diffraction experiments. As found, biological units may be recovered at 80-90% success rate, which makes X-ray crystallography an important source of experimental data on macromolecular complexes and protein-protein interactions. The method is implemented as a public WWW service.