The Interplay between HGF/c-met Axis and Nox4 in BRAF Mutated Melanoma

Background: Melanoma is the leading cause of death due to cutaneous malignancy and its incidence is on the rise. Several signaling pathways, including receptor tyrosine kinases, have a role in the development and progression of melanocytic lesions and malignant melanoma. Among those, the hepatocyte growth factor (HGF)/c-met axis is emerging as a critical player because it can play a role in drug resistance. Indeed, 50% of melanoma patients present BRAF mutations, however, all responders develop resistance to the inhibitors typically within one year of treatment. Interestingly, BRAF inhibitors induce reactive oxygen species (ROS) in melanoma cells, therefore, the aim of this study was to investigate a possible interplay between HGF/c-met and ROS sources, such as NADPH oxidases (Nox). Methods: The expression of c-met and Nox were quantified in 60 patients with primary cutaneous melanoma. In vitro experiments on melanoma primary cells and the cell line were performed to dissect the underpinned molecular mechanism. Results: The outcome of interest was the correlation between the high positivity for both Nox4 and c-met and metastasis occurring at least 1 year later than melanoma diagnosis in BRAF mutated patients, in contrast to nonmutated. In vitro experiments demonstrated that the axis HGF/c-met/Nox4/ROS triggers the epithelial-mesenchymal transition. Conclusions: The observed correlation suggests an interplay between c-met and Nox4 in promoting the onset of metastasis. This study suggests that Nox4 inhibitors could be associated to the current therapy used to treat melanoma patients with BRAF mutations.     

Detection of low‐quality extra virgin olive oils by fatty acid alkyl esters evaluation: a preliminary and fast mid‐infrared spectroscopy discrimination by a chemometric approach

A set of eighty‐one extra virgin olive oils (EVOOs) was analysed according to the new quality parameters relative to the total amount of methyl and ethyl esters of fatty acids [Σ (FAMEs + FAEEs)] and the ratio between ethyl and methyl esters [ratio of FAEEs/FAMEs (RFF)]. Acquisition of the mid‐infrared spectra was also performed by Fourier Transform Infrared Spectroscopy (FT‐IR). Chemical and spectroscopy data were chemometrically elaborated, and FT‐IR coupled by Partial Least Square (PLS) methodology was developed. Results were statistically similar to official procedure in terms of analytical performance for Σ (FAMEs + FAEEs) and RFF in EVOOs: a good agreement between predicted and actual values on calibration data sets was found (0.98 and 0.83, respectively) and the limit of quantification was low enough (29.3 mg kg^?1) considering the actual limits for Σ (FAMEs+ FAEEs). This new approach, time‐saving and environmentally friendly, can be considered as a useful tool for screening procedures.     

A comparison of silver ion HPLC plus GC with Fourier-transform IR spectroscopy for the determination of trans double bonds in unsaturated fatty acids

A rapid procedure has been developed for the isolation of methyl esters of trans-monoenoic fatty acids by silver ion high-performance liquid chromatography. The combined saturated plus trans-monoene fraction was collected for analysis by gas chromatography and comparison with the composition of the unfractionated sample. The trans-monoene contents of partially hydrogenated vegetable oils, determined in this way, are compared with data obtained by Fourier-transform infrared spectroscopy. Alternative methods of base-line correction are discussed. The infrared technique gave values that were a little higher than the chromatographic ones, presumably because the former also detected trans unsaturation in di- and polyunsaturated fatty acids.     

Phosphatidylinositol 3-kinase is recruited to a specific site in the activated IL-1 receptor I

We have developed an on-line archive of neuronal geometry to encourage the use of realistic dendritic structures in morphometry and for neuronal modeling, located at web address www.neuro.soton.ac.uk. Initially we have included full three-dimensional representations of 87 neurons from the hippocampus, obtained following intracellular staining with biocytin and reconstruction using Neurolucida. The archive system includes a structure editor for correcting any departures from valid branching geometry and which allows simple errors in the digitisation to be corrected. The editor employs a platform-independent file format which enforces the constraints that there should be no isolated branches and no closed loops. It also incorporates software for interconversion between the archive format and those used by various neuronal reconstruction and modelling packages. The raw data from digitisation software can be included in the archive as well as edited reconstructions and any further information available. Cross-referenced tables and indexes are updated automatically and are sorted according to a number of fields including the cell type, contributor, submission date and published reference. Both the archive and the structure editor should facilitate the quantitative use of full three-dimensional reconstructions of neurons from the hippocampus and other brain regions.     

Immunocytochemical evaluation of protein kinase C translocation to the inner nuclear matrix in 3T3 mouse fibroblasts after IGF-I treatment

The complex pathway which links the agonist-cell membrane receptor binding to the response at the genome level involves, among other elements, protein kinase C (PKC). Agonists acting at the cell membrane can affect an autonomous nuclear polyphosphoinositide signaling system inducing an activation of nuclear phosphoinositidase activity and a subsequent translocation of PKC to the nuclear region. The fine localization of PKC has been investigated by means of electron microscopy quantitative immunogold labeling in 3T3 mouse fibroblasts, mitogenically stimulated by IGF-I. The enzyme, which in untreated cells is present in the cytoplasm, except for the organelles, and in the nucleoplasm, after IGF-I treatment is reduced in the cytoplasm and almost doubled in the nucleus. The PKC isoform translocated to the nucleus is the alpha isozyme, which is found not only associated with the nuclear envelope but mainly with the interchromatin domains. By using in situ matrix preparations, PKC appears to be retained at the nuclear matrix level, both at the nuclear lamina and at the inner nuclear matrix, suggesting a direct involvement in the phosphorylation of nuclear proteins which are responsible for the regulation of DNA replication.     

A New Triazole-Thiol Compound Organofunctionalized on the Silica Gel Surface: Chemical Properties and Copper Sorption in Ethanol / Water Media

Silicon - This study describes the preparation and characterization of 3-chloropropyl silica gel with 4-amino-5-(4pyridyl)-4H-1,2,4-triazole-3-thiol (SGA) for copper adsorption in different media....     

Gas chromatographic study on high-temperature thermal degradation products of methyl linoleate hydroperoxides

Chromatographic techniques were used to separate secondary products generated by thermal degradation of methyl linoleate hydroperoxides (MLHP). The MLHP were obtained by oxidation, selected, and concentrated by solid-phase extraction (SPE) and thin-layer chromatography (TLC). The purified MLHP were then thermo-degraded in the gas-chromatographic glass liner and analyzed on-line by gas chromatography-mass spectrometry (GC-MS). The MLHP were also thermodegraded and collected in a short silicic acid-packed column, eluted, separated by TLC, and then analyzed by GC. By considering the elution in TLC, the GC retention times and the GC-MS analyses, it was possible to characterize the mono- and the dioxygenated secondary products, particularly those having a boiling point higher than methyl linoleate. The peaks that corresponded to the mono-oxygenated products (epoxy, hydroxy, and keto) were identified, and, on the basis of their MS spectra, molecular structures were proposed. A specific elution order was suggested for keto derivatives: 9-keto,Δ^10,12- and 13-keto,Δ^9,11-octadecadienoate. The hydroxy derivatives, which show the typical fragmentations of 9-hydroxy,Δ^10,12- and 13-hydroxy,Δ^9,11-octadecadienoate, were also identified. On the other hand, identification of the di-oxygenated compounds was more difficult, and, therefore, it was not possible to indicate each positional isomer; however, their elution order could be epoxy-hydroxy and epoxy-keto derivatives.     

The ultrastructure of the rabbit submandibular gland

The secretory granules of the terminal and pre-terminal tracts of the rabbit submandibular gland display distinct features, while the cell morphology is quite similar. Among these cells, some smaller ones presenting an highly dilated endoplasmic reticulum have been identified in the pre-terminal tracts. Some hypothesis have been formulated concerning the function of these cells.