Age related changes in the thermal transition of Turkey leg flexor tendon collagen

A differential scanning calorimetric, thermogravimetric and electron microscopic investigation has been carried out on the uncalcified areas of turkey leg flexor tendon as a function of age. Rehydrated samples exhibit an increase of thermal stability with age. The H_D values drop from about 11 cal·g^–1 in the first weeks of life down to 7 cal·g^–1 after the 11th week.At about 11 weeks, the collagen fibril diameter distribution passes from unimodal to multimodal. The DSC curves as well as the TG-DTG curves recorded from dried samples do not show any appreciable difference with ageing. The variations in thermal behaviour of rehydrated samples and fibril diameter distribution could be related to modifications in water binding with ageing.The Authors are grateful to Dr. G. Fabris for discussion and help in the selection of the samples. They also wish to thank Mr. G. Pizzuto for excellent technical assistance. The financial support by Consiglio Nazionale delle Ricerche is gratefully acknowledged.     

Extraction methods of red blood cell membrane proteins for Multidimensional Protein Identification Technology (MudPIT) analysis

Since red blood cells (RBCs) lack nuclei and organelles, cell membrane is their main load-bearing component and, according to a dynamic interaction with the cytoskeleton compartment, plays a pivotal role in their functioning. Even if erythrocyte membranes are available in large quantities, the low abundance and the hydrophobic nature of cell membrane proteins complicate their purification and detection by conventional 2D gel-based proteomic approaches. So, in order to increase the efficiency of RBC membrane proteome identification, here we took advantage of a simple and reproducible membrane sub-fractionation method coupled to Multidimensional Protein Identification Technology (MudPIT). In addition, the adoption of a stringent RBC filtration strategy from the whole blood, permitted to remove exhaustively contaminants, such as platelets and white blood cells, and to identify a total of 275 proteins in the three RBC membrane fractions collected and analysed. Finally, by means of software for the elaboration of the great quantity of data obtained and programs for statistical analysis and protein classification, it was possible to determine the validity of the entire system workflow and to assign the proper sub-cellular localization and function for the greatest number of the identified proteins.     

Optical technologies in the CNR telecommunication project

Some new results and Perspectives for optical communications are presented as they emerged from an Italian research program on telecommunications. In particular, the experience gained in the implementation of laboratory prototypes of coherent transmission systems is described, as a first step toward a demonstrator of a coherent multichannel distribution network, which is currently under development. About nonlinear optical effects, several experimentations are illustrated in view of their utilization in all-optical switching applications.     

Analysis of thiols by HPLC after fluorescent prelabelling with 4-(6-methylnaphthalen-2-yl)-4-oxobuten-2-oic acid

Summary The use of 4-(6-methylnaphthalen-2-yl)-4-oxobuten-2-oic acid as a fluorogenic reagent in pre-column derivatization for the high-performance liquid chromatography (HPLC) of biologically important thiols (L-cysteine, glutathione, N-acetylcysteine, homocysteine and mercaptopropionylglycine) was investigated. The aroylacrylic acid reacts selectively and rapidly (15 min. at room temperature) with the thiol compounds to give stable fluorescent adducts which can be separated by reversedphase HPLC and detected fluorometrically (_ex 300nm); _em 445nm). The experimental conditions for the thiol derivatization and chromatographic separation are discussed. Applications to the determination of N-acetylcysteine, mercaptopropionylglycine and cysteine are described.Presented in part at Bononiachem, XVI Congresso Nazionale di Chimica, Bologna, October 9–14, 1988.     

Novel Biologically Inspired Collagen Nanofibers Reconstituted by Electrospinning Method

The possibility to prepare bioinspired collagen nanofibers by electrospinning from aqueous suspension of telopeptide-free collagen molecules avoiding both organic solvents and blends with any synthetic and natural polymers has been investigated. The results have highlighted the need for a basic atmosphere between the needle and the ground collector in order to increase the environmental pH during the collagen molecules self-assembly along the electrostatic force lines. Morphological, spectroscopic and calorimetric analyses carried out on the electrospun collagen nanofibers have opened the possibility to take advantage of this new approach in order to prepare an ideal biomimetic reinforcing component of new biomedical and surgical biomaterials.     

Frequency intermittency and energy pumping by linear attachments

The present paper considers the problem of realizing an effective targeted energy pumping from a linear oscillator to a set of ungrounded linear resonators attached to it. Theoretical as well as numerical results demonstrate the efficacy of using a complex attachment as a passive absorber of broadband energy injected into the primary structure. The paper unveils also the existence of an instantaneous frequency associated with the master response characterized by intermittency: a rather surprising result for a linear autonomous system. Comparison with nonlinear energy sinks demonstrates that the two systems have some analogies in this respect and that the linear complex attachment is a very efficient energy trap.     

Structural and chemical characterization of a cutaneous calcification

Dynamic thermogravimetry, X-ray diffraction, spectroscopic and chemical analyses have been carried out on the cutaneous calcifications present in a case of Porphyria cutanea tarda. The inorganic deposits are constituted by a poor crystalline B carbonated apatite characterized by an almost stoichiometric Ca/P molar ratio, a low magnesium relative content and a high thermal stability. The inorganic crystallites grow in the cutaneous calcification without any preferential orientation in the tissue where the collagen fibrils are isotropically distributed. The results reveal that the cutaneous calcifications display a close similarity with the inorganic deposits isolated from atheromatous plaques of aorta and calcified mitral valves.     

Adsorption and spectroscopic characterization of lactoferrin on hydroxyapatite nanocrystals

Lactoferrin (LF), a well-characterized protein of blood plasma and milk with antioxidant, cariostatic, anticarcinogenic and anti-inflammatory properties, has been adsorbed onto biomimetic hydroxyapatite (HA) nanocrystals at two different pH values (7.4 and 9.0). The interaction was herein investigated by spectroscopic, thermal and microscopic techniques. The positive electrostatic surface potential of LF at pH 7.4 allows a strong surface interaction with the slightly negative HA nanocrystals and avoids the protein-protein interaction, leading to the formation of a coating protein monolayer. In contrast, at pH 9.0 the surface potential of LF is a mix of negative and positive zones favouring the protein-protein interaction and reducing the interaction with HA nanocrystals; as a result a double layer of coating protein was formed. These experimental findings are supported by the good fittings of the adsorption isotherms by different theoretical models according to Langmuir, Freundlich and Langmuir-Freundlich models. The nanosized HA does not appreciably affect the conformation of the adsorbed protein. In fact, using FT-Raman and FT-IR, we found that after adsorption the protein was only slightly unfolded with a small fraction of the α-helix structure being converted into turn, while the β-sheet content remained almost unchanged. The bioactive surface of HA functionalized with LF could be utilized to improve the material performance towards the biological environment for biomedical applications.