On the corrosion, stress corrosion and cytocompatibility performances of ALD TiO2 and ZrO2 coated magnesium alloys.

Magnesium alloys are increasingly studied as materials for temporary implants. However, their high corrosion rate and susceptibility to corrosion-assisted cracking phenomena, such as stress corrosion cracking (SCC), continue to prevent their mainstream use. Recently, coatings have been considered to provide an effective solution to these issues and researchers have focused their attention on Atomic Layer Deposition (ALD). ALD stands out as a coating technology due to the outstanding film conformality and density achievable, and has shown encouraging preliminary results in terms of reduced corrosion rate and reduced SCC susceptibility. Here, we contribute to the ongoing interest in ALD-coated Mg alloys, providing a comprehensive characterisation of the effect of 100 nm thick ALD TiO2 and ZrO2 coatings on the corrosion behaviour and SCC susceptibility of AZ31 alloy. Moreover, we also investigate the effect of these coatings on the induced biological response. Our results suggest that the ALD coatings can improve the corrosion and SCC resistance of the Mg alloy, with the ZrO2 ALD coating showing the best improvements. We suggest that the different corrosion behaviours are the cause of the cytocompatibility results (only the ZrO2 ALD coating was found to meet the demands for cellular applications). Finally, we leverage on considerations about the coatings' wettability, electrochemical stability and surface integrity to justify the different results.

Improving stress corrosion cracking behavior of AZ31 alloy with conformal thin titania and zirconia coatings for biomedical applications.

Magnesium and its alloys have been widely studied as materials for temporary implant devices. However, corrosion-assisted cracking phenomena such as stress corrosion cracking (SCC) continue to prevent their mainstream use. For the first time, we explore the SCC susceptibility of Atomic Layer Deposition (ALD) coated AZ31 alloys in Simulated Body Fluid (SBF). Conformal 100 nm coatings of titania and zirconia were deposited on standard dogbone specimens and subjected to slow strain rate tests at 3.5 10-6 s-1 and a temperature of 37 °C. Remarkably, the SCC susceptibility index IUTS was reduced by 6% and 40% and the Iε was reduced by more than 70% and 76% with a titania and zirconia coating, respectively. Potentiodynamic polarization, hydrogen evolution and fracture behavior of the samples revealed the drastic corrosion reduction to be the main reason for the susceptibility reduction. We discuss the observed SCC behavior of our samples in light of the coatings' electrochemical activities, wettabilities, surface integrities and mechanical properties. This straightforward conformal surface treatment can be useful as a workaround for one of the major bottlenecks of biomedical Mg based implants and hence provides a possible pathway for making them more commonplace in the field.

Enhancement of stress corrosion cracking of AZ31 magnesium alloy in simulated body fluid thanks to cryogenic machining.

Magnesium and its alloys have recently attracted great attention as potential materials for the manufacture of biodegradable implants. Unfortunately, their inadequate resistance to the simultaneous action of corrosion and mechanical stresses in the human body have hampered their use as implant materials. This work aims at evaluating the Stress Corrosion Cracking (SCC) susceptibility of the AZ31 Mg alloy after being machined under cryogenic cooling. The SCC behaviour was evaluated by means of Slow Strain Rate Tests (SSRTs) in Simulated Body Fluid (SBF) at 37 °C. Prior to testing, a full characterization of the machined surface integrity, including microstructural observations, residual stress, nano-hardness measurements and surface texture analysis was carried out together with the assessment of the corrosion properties through potentiodynamic polarization curves. In addition, the morphology of the fracture surfaces after SSRTs was analysed by means of 3D optical profiler and Scanning Electron Microscopy (SEM). The improved corrosion resistance due to the increased extension of the nano-surface layer and to the compressive residual stresses represents the reason of the reduced SCC susceptibility of cryogenically machined AZ31 samples as compared to dry machined ones.

Consequences of heat shock protein 72 (Hsp72) expression and activity on stress-induced apoptosis in CD30+ NPM-ALK+ anaplastic large-cell lymphomas.

Understanding the mechanisms that control stress-induced apoptosis is critical to explain how tumours respond to treatment, as cancer cells frequently escape drug toxicity by regulating stress response through heat shock protein (HSP) expression. The overexpression of Hsp72, in particular, results in increased incidence of cell transformation, and correlates with poor prognosis in a wide range of cancers. We have shown that Hsp72 assists folding of oncogenic NPM-ALK kinase in anaplastic large-cell lymphomas (ALCLs), but its role in the maintenance of the malignant phenotype remains uncertain. Therefore, we assessed Hsp72 expression in ALCLs, investigating more in detail the mechanisms that regulate its status and activity. We found that Hsp72 is unique among the HSPs involved in tumourigenesis to be overexpressed in ALK(+) tumours and cell lines and to be induced by stress. Different from other HSPs, Hsp72 prevents cell injury, Bax activation and death by apoptosis in ALK(+) cells, acting both upstream and downstream of mitochondria. Conversely, Hsp72 is underexpressed in ALK(-) ALCL cells, and it is unable to protect cells from apoptosis under stress. Moreover, when Hsp72 expression is reduced following NPM-ALK inhibition, lymphoma cells undergo apoptosis, demonstrating the importance of Hsp72 in regulating ALCL stress response and drug sensitivity.

Chronological ageing of human hair keratin fibres.

Examination of very long hair (length > 2.4 m) using a large range of evaluation methods including physical, chemical, biochemical and microscopic techniques has enabled to attain a detailed understanding of natural ageing of human hair keratin fibres. Scrutinizing hair that has undergone little or no oxidative aggression--because of the absence of action of chemical agents such as bleaching or dyeing--from the root to the tip shows the deterioration process, which gradually takes place from the outside to the inside of the hair shaft: first, a progressive abrasion of the cuticle, whilst the cortex structure remains unaltered, is evidenced along a length of roughly 1 m onwards together with constant shine, hydrophobicity and friction characteristics. Further along the fibre, a significant damage to cuticle scales occurs, which correlates well with ceramides and 18-Methyl Eicosanoic Acid (18-MEA) decline, and progressive decrease in keratin-associated protein content. Most physical descriptors of mechanical and optical properties decay significantly. This detailed description of natural ageing of human hair fibres by a fine analysis of hair components and physical parameters in relationship with cosmetic characteristics provides a time-dependent 'damage scale' of human hair, which may help in designing new targeted hair care formulations.

Characterisation and distribution of calcitonin gene-related peptide in a primitive teleost, the eel, Anguilla anguilla and comparison with calcitonin.

Radioimmunoassay (RIA), radioreceptor assay and chromatography were used to study the occurrence of calcitonin gene-related peptide (CGRP) in a primitive teleost, the eel, Anguilla anguilla. Immunologically and biologically active CGRP-like molecules were found in brain, heart, kidney, liver, spleen and ultimobranchial body with the higher concentrations in brain, spleen and heart. Gel exclusion chromatography of heart and spleen extracts followed by SDS-PAGE showed that the eel CGRP-like molecules presented a molecular weight between 3.30 and 3.95 kDa similar to that of human CGRP. The wide distribution of CGRP reflects its multiple role as brain neuromediator and peripheral paracrine effector as described in mammals. In comparison, the distribution of calcitonin (CT) was much more restricted, immunologically and biologically active CT-like molecules being localised in the ultimobranchial bodies (UBB) that is the site of CT synthesis in non-mammalian vertebrates. In plasma, CGRP-like concentrations were 10 to 100 higher than those of CT. These high concentrations in a primitive teleost strengthen the possible endocrine role of CGRP in early vertebrates and emphasise the important role of this hormone in evolution.

Biomineralisation markers during a phase of active growth in Pinctada margaritifera.

To search for the biochemical parameters involved in calcium and carbonate transport during crystal formation and biomineralisation in nacreous molluscs, the carbonic anhydrase activity, the levels of calciotropic hormones in hemolymph and in tissues and the circulating concentration of calcium were measured in pearl oysters (Pinctada margaritifera) during a phase of active growth. Activity of carbonic anhydrase in gill tissue increased linearly with age of the animals, while no age variation in activity was noted for the mantle. The circulating level of total calcium increased during the growth of the animals. Calciotropic hormones were radioimmunoassayed in gill, mantle and hemolymph. Only a calcitonin-gene related peptide (CGRP) could be detected and its concentration decreased as a function of growth, both in hemolymph and mantle. No variation in CGRP concentration with age was observed in gill tissue. Our data demonstrate that carbonic anhydrase and a molecule biologically and immunologically related to CGRP are involved during growth of the animals. In addition, this study shows the presence of three main calcium compartments, gill, hemolymph and mantle, involved in the biomineralisation process.

[Beating heart in "aortic no-touch off-pump" techniques: a simple answer to complex surgical situations]. / Le coeur battant en "no-touch aortique complet": une solution simple à des problémes chirurgicaux complexes.

Three cases reports illustrate the concept of beating heart cardiac surgery and its advantages regarding the prevention of arterial emboli.

TEM-89 beta-lactamase produced by a Proteus mirabilis clinical isolate: new complex mutant (CMT 3) with mutations in both TEM-59 (IRT-17) and TEM-3.

TEM-89 (CMT-3) is the first complex mutant beta-lactamase produced by a clinical strain of Proteus mirabilis (strain Pm 631). This new enzyme, which has a pI of 6.28, is derived from TEM-3 and has a single amino acid substitution also encountered in TEM-59 (inhibitor-resistant TEM beta-lactamase IRT-17): Ser-130 to Gly. TEM-89 hydrolyzed penicillins to the same extent that TEM-3 did but lost almost all hydrolytic activity for cephalosporins and, like TEM-59, was highly resistant to inhibitors.

Effect of calcitonin gene related peptide on the adenylate cyclase activity in abalone gill membranes.

The aim of the present study was to investigate and to compare the effect of calciotropic hormones, human calcitonin gene related peptide (CGRP) I and II, salmon calcitonin (CT) and human amylin on the adenylate cyclase activity in abalone gill membranes. In addition to human CGRPI, human CGRPII and salmon CT stimulated the adenylate cyclase activity. No effect was observed with amylin. The higher effect was observed with human CGRPI and II that induced a 160-170% increase of the enzyme activity. Fifty percent of the maximal activity was observed with 3 and 8 nM of CGRP I and II, respectively. Salmon CT induced a lower effect: the maximal activity was obtained with a hormone concentration of 266 nM and represented a 130% stimulation of the basal activity. In the presence of CGRP 8-37, a competitive antagonist of CGRP action, the stimulation observed with CGRPI was abolished and returned to the basal level. This study points out that, in invertebrates, CGRP receptors present in gill membranes are linked to an adenylate cyclase system similar to that described in vertebrates. In addition, these data are in favour of a role for CGRP in branchial function both in non-mammalian vertebrates and invertebrates

Interaction of cysteine proteases with calciotropic hormone receptors.

The effect of two cysteine proteases: papain and a cathepsin L-like enzyme purified from the oesophagus of Nephrops norvegicus (NCP) was studied on the specific binding of calcitonin (CT) and calcitonin gene related peptide (CGRP) to rat kidney and liver membranes, respectively. In addition, the response of adenylyl cyclase to increasing concentrations of these two enzymes was investigated. Each protease inhibited the initial CGRP and CT binding to rat liver and kidney membranes, respectively, in a manner not significantly different from that obtained in the presence of the unlabeled standard. The adenylyl cyclase activity in rat liver membranes was increased by the addition of each enzyme. The response was higher with papain that induced a fivefold increase of enzyme activity at a 4-microg/ml enzyme concentration. In rat kidney membranes, the magnitude of the response was identical with both enzymes. In contrast with NCP, papain induced a biphasic response. Leupeptin and E(64), two specific inhibitors of cysteine proteases, reversed the observed effects. Trypsin induced an inhibition of the liver membrane adenylyl cyclase activity and an activation in rat kidney membranes at low protease concentration. Thus, cysteine proteases are able to act, in vitro, at the receptor level in target organs specific for calciotropic hormones.

Characterization of binding sites for calcitonin gene related peptide in abalone gill.

Target organs for calcitonin gene related peptide were investigated in the abalone. To elucidate the function of this neuropeptide in the biomineralization process, we have localized, in different tissues from abalone, specific binding sites for human calcitonin gene related peptide (hCGRP). Highest binding was observed in gill membranes where two classes of affinity components were identified. The affinity constants and the number of binding sites per mg of proteins for the site I were 5 X 10(9)M(-1)and 8.8 x 10(10). For the site II, the affinity constant was 1.34 X 10(7) M(-1)and the number of binding sites per mg of proteins was 1.1 x 10(12). In contrast, no specific calcitonin binding could be detected in every tested tissue, but the similar displacement of the 125I-labeled CGRP binding with unlabeled hCGRP and sCT suggested that, in the abalone, the identified receptor could belong to a third class receptor subtype, that recognize both hCGRPalpha and sCT. These data suggest that, during evolution, the role of CGRP in gill function is particularly well conserved and that this neuropeptide is likely to participate in the control of hydromineral metabolism in aquatic species.

Isolation of an acid fraction from a fish protein hydrolysate with a calcitonin-gene-related-peptide-like biological activity.

The possibility of obtaining calcitonin and/or calcitonin-gene-related peptide (CGRP) immunorelated molecules from partly digested proteins was investigated with fish and shrimp hydrolysates. These two peptides were quantified by both radioimmunoassay and radioreceptor assay; the positive extracts were partly purified. Different hydrolysates were analysed: cod head, stomach and viscera hydrolysates, a shrimp hydrolysate and two sardine hydrolysates. Although each cod extract interacted in the CGRP radioimmunoassay, none of these extracts was able to displace the CT binding to its antibody. In contrast, shrimp and sardine hydrolysates interacted with both radioimmunoassays. Radioreceptor assays performed on the same extracts demonstrated that only three extracts contained the structural determinants that allowed them to interact in the CGRP radioreceptor assay. No interaction with the calcitonin radioreceptor assay could be demonstrated. Molecular sieving of the two sardine extracts showed that the immunoreactivity was resolved into two main fractions. The higher-molecular-mass fraction interacted only in the CGRP radioreceptor assay. The results obtained suggest the presence of a biologically related CGRP molecule in peptone hydrolysates and requires further investigation into the role of these peptide fragments in the regulation of intestinal function by partly digested proteins.

CGRP-like molecules and carbonic anhydrase activity during the growth of Pecten maximus.

In order to investigate the physiological role of calcitonin gene-related peptide (CGRP) in mollusc, both circulating CGRP-related molecules and gill or mantle carbonic anhydrase activity were analysed during the annual growth of Pecten maximus. CGRP like molecules measured by radioreceptor assay increased significantly during the annual cycle. Similarly, gill carbonic anhydrase activity increased and showed a maximum activity when growth is stimulated to the greatest extent. Correlation studies showed a significant relationship between the tissue weight and either the gill carbonic anhydrase activity or the CGRP-related molecules determined by radioreceptorassay. This observation suggests a possible interaction between carbonic anhydrase activity and CGRP. Accordingly, we searched for a direct effect of CGRP on the gill carbonic anhydrase activity. In gill membranes, CGRP stimulated the carbonic anhydrase activity. The maximum effect was obtained at a CGRP concentration of 50 nM.

Characterization of a calcitonin gene related peptide-like molecule in the abalone, Haliotis tuberculata.

Immunoreactive related CGRP molecules (ir-CGRP) were identified in the abalone, Haliotis tuberculata, mainly in mantle and cephalic part extracts. Ir-CGRP in both tissues accounted for 461 and 455.6 pg per mg of proteins, respectively. These CGRP-immunoreactive molecules were further analyzed for their ability to interact with the CGRP radioreceptor assay. In specific target tissues for CGRP (rat liver membranes), 50% inhibition of 125I-labeled CGRP specific binding was observed with 4.7 micrograms and 21.1 micrograms of proteins from mantle and cephalic part extract, respectively. These molecules were submitted to gel-filtration chromatography on a Sephacryl S-100 column and were further analyzed in the radioreceptor assay specific for CGRP. The elution position of these molecules suggested a molecular weight close to that of synthetic salmon calcitonin.

A lobster cysteine protease with immunoreactivity and activities of calcitonin and CGRP.

The high concentrations of molecules immunologically related to salmon calcitonin (CT) and/or to human calcitonin gene-related peptide (CGRP) in the oesophagus of the norway lobster Nephrops norvegicus have been examined. In the present study. We report the purification of these molecules by means of a specific radioimmunoassay for calcitonin and calcitonin gene related peptide. The immunoreactive molecules were tested for their functional similarities with CT and CGRP. This was investigated by measuring their ability to interact with CGRP and CT radioreceptor assays and to stimulate the adenylate cyclase activity in rat liver and kidney membranes, respectively. In addition, the purified product was injected in young rats in order to check for a CT-like biological activity of these molecules. The combination of these tests led us to purify a molecular form of 33 kDa. N-terminal sequence analysis of this protein revealed a considerable homology with the lobster cysteine proteases and the human cathepsin L. Control experiments performed with the highly purified American lobster cysteine protease I showed that crustacean cysteine proteases given in vivo to rats induce a fall in the plasma calcium and phosphate levels. This study therefore adds further documentation for a common ancestral origin of CT, CGRP and the much large cysteine proteases from invertebrates.

Characterization of binding sites for calcitonin gene-related peptide in the mollusc gill.

Target organs for calcitonin gene-related peptide (CGRP) were investigated in Pecten maximus using 125I-labelled human CGRP. CGRP was shown to interact specifically with mantle and gill tissue. Receptor studies using branchial membrane preparations indicated that the binding was time dependent. Scatchard analysis of binding data showed that there was a single class of binding sites. The affinity constant was found to be 0.7.10(8) M-1 and the number of binding sites 2600.10(8)/mg of protein. Salmon CT inhibited the binding of 125I-labelled CGRP to branchial membranes with a lesser efficiency than that of the unlabelled hormone. A 40% inhibition of the 125I-labelled CGRP binding was observed in the presence of 2.6 and 26 nM CGRP and salmon CT, respectively. In addition, 200 nM human CGRP inhibited 25 and 10% of the basal branchial and mantle adenylate cyclase activity, respectively. These data suggest that CGRP participates in the regulation of the branchial function in molluscs probably via a vasoconstrictor role.

[Value of ilio-hypogastric block in appendectomy in children]. / Intérêt du bloc ilio-hypogastrique pour appendicectomie chez l'enfant.

This prospective study aimed to evaluate the efficiency of ilio-hypogastric nerve block for control of post appendicectomy pain in children. Forty-two children aged 3-15 years scheduled for appendicectomy were anaesthetized in the same way. After randomization, a preoperative ilio-hypogastric nerve block was performed in 21 patients. Twenty one were not blocked. The postoperative pain assessment showed a better analgesia in the blocked children group. Analgics were required less in group. Five inefficient blocks were recorded. No complications were noted. Ilio-hypogastric block was found to be safe and efficient for control of post-appendicectomy pain in most children.

Epithelial ovarian tumors in young women under 35 years.

The aim of this study was to investigate the clinical-pathological pattern of primary malignant epithelial ovarian tumors in women under 35 years old. We have retrospectively analyzed data from 545 patients with these tumors, identifying 49 patients under age 35. We found a higher frequency of borderline tumors and early-stage tumors in young women than in those over 35. The influence of age as a factor in survival was evaluated by comparing the outcome in young patients and in women over 35. The 5-year survival rate, according to stage and aggressiveness factors, was significantly better in young patients. These findings support the concept of a preclinical phase of epithelial carcinoma and show that young women may be selected for conservative surgery, allowing a good quality of life.

Calcitonin gene-related peptide stimulates carbonic anhydrase activity in trout gill membranes.

The effect of human calcitonin gene-related peptide (hCGRP) on carbonic anhydrase activity in trout branchial membranes was investigated. hCGRP stimulated the carbonic anhydrase activity, 5-fold over the basal value. This effect was specific: in the presence of sCT only a 2.4-fold increase was observed. Rat amylin that has a 43% homology with hCGRP had no effect on carbonic anhydrase activity. On the other hand hCGRP(8-37), which is a partial antagonist, inhibited the hCGRP-stimulated carbonic anhydrase activity. This effect was observed at low hormonal concentration and suggests the involvement of hCGRP in regulating carbonic anhydrase activity in this important target organ for CO2 exchange.