The use of alternative systems for the ecotoxicological screening of complex mixtures on fish populations

This paper presents the results of the use of alternative systems in a screening study of four complex mixtures. The following tests were performed: in vitro induction of micronuclei in a rainbow trout-derived cell line by flow cytometry, and hatching percentage, time of hatching and teratogenic alterations on the embryolarval development on medaka fish eggs. The results obtained with the proposed tests in this study allows an increase in the information level in a short period of time (2 weeks), using very low sample volumes (< 100 ml). Inclusion of chronic and specific effects (genotoxicity and teratogenicity) allows the selection of the most sensitive endpoint to increase security factors in the ecotoxicological assessment of complex mixtures, so that detailed studies can be focused only on those samples which require further research.     

Experimental study and finite element analysis of the elastic instability of composite lattice structures for aeronautic applications

The work presented in this paper is focused on the finite element modelling (FEM) of the failure behaviour of lattice composite hollow structures that have been subjected to an external hydrostatic pressure. Furthermore, the development of an experimental procedure to measure the aforementioned resistance, and to test the FEM model is also presented. Carbon fibres composite hollow cylinders with a lattice structure and with different geometries were produced and tested. In order to develop a design tools for such structures, all the experiments performed were computer simulated using finite element modelling. The results obtained with FEM simulation provide further insight to analyze and investigate the failure mechanism. The elastic instability of the studied structures was therefore analyzed and the influence of element geometry on the collapsing resistance thereof was considered. As a result of the study it has been possible to locate three different failure modes which where strictly related to the length of the cylinders. Both the shape of the broken tube and the lever of failure stresses were correctly predicted by the FEM model.     

Plasma vitellogenin levels and gonadal morphology of wild carp (Cyprinus carpio L.) in a receiving rivers downstream of sewage treatment plants

A field study was carried out in order to assess, in a wild cyprinid fish, the biological effects related to the exposure to estrogenic compounds. The gonadosomic index (GSI), plasma vitellogenin (VTG) level and histological observations of gonads were performed to evaluate the reproductive health in common carp (Cyprinus carpio) from three rivers receiving sewage treatment plant (STP) effluents (Guadarrama, Henares and Jarama rivers, Spain) and from a reference site (Lozoya river). No significative differences in male GSI between sites were described. Only in the 18% of fish from Guadarrama river were elevated plasma VTG levels detected. The presence of oocytes in the testis was not found in any of males examined. The results suggest that the released estrogenic compounds by STPs effluents are not enough capable to adversely affect the gonadal development of the fish studied. Additional studies will be required to assess the risk of estrogenic compounds on fish from other sites of this area.     

Short-term transplantation of isolated human ovarian follicles and cortical tissue into nude mice

This study was designed to evaluate follicular survival and growth after short-term transplantation of fresh isolated human follicles and ovarian cortical tissue to nude mice. Ovarian biopsies were obtained from nine women undergoing laparoscopy. Twelve nude mice were xenografted with an ovarian cortical fragment in the right ovarian bursa, and a clot containing isolated follicles in the left, for a period of 7 days. One ungrafted fragment was used as a control. Histological sections were analyzed to determine follicle number and stage. The proliferative status of follicular cells was assessed by Ki-67 immunostaining. A total of 659 follicles was analyzed by histology and 545 follicles by immunohistochemistry. The percentage of primordial follicles was found to be markedly reduced 1 week post-grafting when compared with ungrafted tissue, while the percentage of primary follicles had significantly increased. Only 8% of follicles showed Ki-67-positive granulosa cells before grafting, whereas 1 week after grafting, 71% of follicles in fragments and 67% of isolated follicles were Ki-67-positive (P<0.001). Moreover, the histological aspect of isolated follicle grafts was similar to that of grafted fragments: follicles were surrounded by vimentin-positive stroma-like tissue of human origin, as confirmed by fluorescent in situ hybridization with human-specific probes. Our results demonstrate, for the first time, that isolated human follicles are able to survive and grow after xenografting. This study also shows massive in vivo follicular activation after transplantation of grafted fragments and isolated follicles. One week after grafting, well-structured stroma-like tissue of human origin was observed around the isolated follicles. The potential origin of this stroma is discussed.     

Recent progress on the study of the microstructure and mechanical properties of ECAE copper

Results on the microstructure and the tensile properties of equal channel angular extruded (ECAE) copper processed for one to 16 passes are presented and compared with the available literature data. With increasing number of passes (N), the microstructure changes from a strongly elongated shear band structure after N = 1 and 2, towards a more equiaxed subgrain and grain structure. This is accompanied by a decrease in the cell wall or subgrain-boundary widths and an increase in recovered or even recrystallised grain structures with low dislocation densities. Electron backscatter diffraction measurements have indicated that for lower N, the location of Σ3 boundaries is restricted to shear bands, while at greater N, Σ3 boundaries were found to be more widely distributed. Texture measurements indicate close similarity with simple shear texture components and a spread of the orientation components with greater N. Upon comparing the tensile behaviour of as-ECAE Cu with the surveyed literature, broad agreement on the strength of the material is achieved. However, a strong variation in the percentage elongation to failure is also noted. Strain hardening and deformation kinetic analysis via strain rate jump tests indicate an evolution from stage III to V hardening during post-ECAE compression and a saturation in the strain rate sensitivity after N = 4 resulting in maximum values of ∼0.02. Our results suggest that rather than a change in deformation mechanism, the increase in ductility with increasing N is associated with an increase in the mean free path of dislocations—with the grain boundaries remaining actively involved as the transmitter of plastic strain and their interaction with dislocations being the rate controlling deformation mechanism.

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Keratins extracted from Merino wool and Brown Alpaca fibres as potential fillers for PLLA-based biocomposites

This paper reports on the promising perspectives of using keratins extracted by sulphitolysis reaction from Merino wool (KM) and Brown Alpaca fibres (KA) in poly (l-lactide) (PLLA)-based biomaterials. Both types of keratin were dispersed in chloroform (CHCl₃) and tetrahydrofuran (THF), and optimisation of dispersion methods and parameters using the two selected solvents was considered. The extraction yield, as well as supermolecular structures, morphology and thermal behaviour of the two proteins before and after the regeneration in CHCl₃ was investigated. The results indicated that the supermolecular structures and thermal behaviour of the two proteins were affected by the interaction with CHCl₃, producing decrease of the amount of α-helix structures in KM and an increase for KA, a slight decrease of β-sheet structures and a reduced thermal stability of α-crystallites for both keratins. Biocomposite films based on PLLA polymer matrix and two different contents of Merino wool and Brown Alpaca keratins (1 % and 5 % wt) were successfully developed by solvent casting in chloroform and the resulting morphologies after incorporation of different keratins (as a function of content and source) give evidence of different surface topographies, with a random distribution of keratin in flask-like structure. PLLA/5KA and PLLA/5KM samples with 1 % and 5 % wt of keratins show a specific pore-like surface microstructure, induced by solvent evaporation.     

The role of the interphase on the shear induced failure of multiwall carbon nanotubes reinforced epoxy nanocomposites

Multiwall Carbon Nanotubes (MWCNT) with an elevated aspect ratio were chemically functionalized with amines and two types of epoxide groups. Thermogravimetric analysis and Fourier Transform‐Infrared Spectroscopy (FTIR) analysis corroborated that the functionalization degree was substantial (up to 30 wt %) and the presence of a covalent bond with the MWCNT. The functionalized MWCNT (f‐CNT) were incorporated into an epoxy matrix after its dispersion in the diglycidyl ether of bisphenol A (DGEBA) precursor. To induce a shear failure mode, a short‐beam (SB) experimental setup was implemented. The SB shear strength (SBSS) proved that the functionalization had a strong influence on its value. For the case of pristine CNT, a neutral effect was obtained. A strong detrimental effect (?17.2% ± 9.5) was measured for the amine type f‐CNT and a positive effect (up to 10.9% ± 8.9) was measured of the epoxide type f‐CNT. Fractographic analysis of each formulation was correlated with SBSS performance, proving that the surface texture of the fractured samples was strongly correlated to its value. Furthermore, dynamic mechanical analysis proved that the damping factor and the crosslink molecular weight were correlated with the SBSS performance. A lower full width at half maximum of the damping factor was associated to an improvement of SBSS. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 41364.     

Efficacy of chlorhexidine as a postmilking teat disinfectant for the prevention of bovine mastitis during lactation

A natural exposure trial was conducted for 12 mo in a herd of 150 lactating Jersey cows to determine efficacy of a .35% chlorhexidine teat dip containing a glycerine emollient for the prevention of bovine intramammary infections. Right teats of cows were dipped in the experimental teat dip after milking machine removal and left teats were not dipped. The herd was free of Streptococcus agalactiae and had a low prevalence of Staphylococcus aureus. Most new major pathogen intramammary infections resulted from Streptococcus species, primarily Streptococcus uberis and Streptococcus dysgalactiae. New infections by Streptococcus species were significantly lower in teats dipped in chlorhexidine than in undipped teats. Overall efficacy of the chlorhexidine teat dip against major mastitis pathogens was 50%. The experimental teat dip also reduced coagulase-negative Staphylococcus species infections 49.0% and Corynebacterium bovis infections 65.2%. Overall efficacy against minor mastitis pathogens was 54.0%. No irritation or chapping of teats dipped in the experimental teat dip was observed.     

Dose effect on the uptake and accumulation of hydroxytyrosol and its metabolites in target tissues in rats

Hydroxytyrosol (HT) is the most prominent phenolic compound of virgin olive oil and due to its scientifically validated biological activities it is entering to the market as a potentially useful supplement for cardiovascular disease prevention. The aim of the present study was to investigate the relationship between the HT dose intake and its tissue uptake in rats, and thus, providing complementary information in relation to the target‐dose relationship. Rats were given a refined olive oil enriched with HT at different doses (1, 10, and 100 mg/kg) and they were sacrificed after 5 h to ensure the cell tissue uptake of HT and its metabolites. Plasma samples and different organs as liver, kidney, heart and brain were obtained, and HT metabolites were analyzed by UPLC‐MS/MS. The results showed that HT and its metabolites could be accumulated in a dose‐dependent manner basically in the liver, kidney, and brain and were detected in these tissues even at nutritionally relevant human doses. The detection of free HT in liver and kidney was noteworthy. To date, this appears to be the only biologically active form, and thus, it provides relevant information for optimizing the potential applications of HT to prevent certain hepatic and renal diseases. In recent years, HT and its derivatives have led to a great interest from the virgin olive oil producers and manufacturers of nutraceutical supplements. The increasing interest in HT is mainly due to the European Food Safety Agency (EFSA) Panel on Dietetic Products, Nutrition, and Allergies (NDA) scientific opinion that established a cause‐and‐effect relationship between the consumption of olive oil polyphenols and protection of LDL particles from oxidative damage 1 . Based on this positive opinion, the health claim “Olive oil polyphenols contribute to the protection of blood lipids from oxidative stress” was included in the list of health claims 2 , being the only authorized health claim in the European Union regarding polyphenols and health.     

Prevalence and behaviour of Bacillus cereus in a REPFED of Italian origin

This paper reports on the prevalence and behaviour of Bacillus cereus in gnocchi, a REPFED of Italian origin. A survey of gnocchi under varying storage conditions, revealed that, although B. cereus was found in 33% of the samples, the contamination level was lower than 10(2) CFU/g for the unstored and the refrigerated (8 degrees C) samples. Counts increased with increasing storage temperatures and prolonged storage times in samples prepared without sorbic acid. The effect of different formulations (sorbic, citric and lactic acid) and storage conditions (8, 12 and 20 degrees C) have been evaluated in a challenge testing with spores of B. cereus. Results indicate that the use of sorbic acid in association with citric or lactic acid to pH 5.0 is effective in inhibiting growth of B. cereus and the anticipated shelf life of the product is safe even if temperature abuse occurs. If sorbic acid is omitted, lactic acid can inhibit B. cereus growth during storage at 8 degrees C. On the contrary, when temperature abuse occurs (12 and 20 degrees C), lactic or citric acid are not able to prevent growth of B. cereus.