Melanin Distribution in Human Skin: Influence of Cytoskeletal,Polarity, and Centrosome-Related Machinery of Stratum basale Keratinocytes

Melanin granules cluster within supra-nuclear caps in basal keratinocytes (KCs) of the human epidermis, where they protect KC genomic DNA against ultraviolet radiation (UVR) damage. While much is known about melanogenesis in melanocytes (MCs) and a moderate amount about melanin transfer from MC to KC, we know little about the fate of melanin once inside KCs. We recently reported that melanin fate in progenitor KCs is regulated by rare asymmetric organelle movement during mitosis. Here, we explore the role of actin, microtubules, and centrosome-associated machinery in distributing melanin within KCs. Short-term cultures of human skin explants were treated with cytochalasin-B and nocodazole to target actin filaments and microtubules, respectively. Treatment effects on melanin distribution were assessed by the Warthin–Starry stain, on centrosome-associated proteins by immunofluorescence microscopy, and on co-localisation with melanin granules by brightfield microscopy. Cytochalasin-B treatment disassembled supra-nuclear melanin caps, while nocodazole treatment moved melanin from the apical to basal KC domain. Centrosome and centriolar satellite-associated proteins showed a high degree of co-localisation with melanin. Thus, once melanin granules are transferred to KCs, their preferred apical distribution appears to be facilitated by coordinated movement of centrosomes and centriolar satellites. This mechanism may control melanin’s strategic position within UVR-exposed KCs.     

Cardioinotropic Effects in Subchronic Intoxication of Rats with Lead and/or Cadmium Oxide Nanoparticles

Subchronic intoxication was induced in outbred male rats by repeated intraperitoneal injections with lead oxide (PbO) and/or cadmium oxide (CdO) nanoparticles (NPs) 3 times a week during 6 weeks for the purpose of examining its effects on the contractile characteristics of isolated right ventricle trabeculae and papillary muscles in isometric and afterload contractions. Isolated and combined intoxication with these NPs was observed to reduce the mechanical work produced by both types of myocardial preparation. Using the in vitro motility assay, we showed that the sliding velocity of regulated thin filaments drops under both isolated and combined intoxication with CdO–NP and PbO–NP. These results correlate with a shift in the expression of myosin heavy chain (MHC) isoforms towards slowly cycling β–MHC. The type of CdO–NP + PbO–NP combined cardiotoxicity depends on the effect of the toxic impact, the extent of this effect, the ratio of toxicant doses, and the degree of stretching of cardiomyocytes and muscle type studied. Some indices of combined Pb–NP and CdO–NP cardiotoxicity and general toxicity (genotoxicity included) became fully or partly normalized if intoxication developed against background administration of a bioprotective complex.     

Bone Marrow-Mesenchymal Stromal Cell Secretome as Conditioned Medium Relieves Experimental Skeletal Muscle Damage Induced by Ex Vivo Eccentric Contraction

Bone marrow-mesenchymal stem/stromal cells (MSCs) may offer promise for skeletal muscle repair/regeneration. Growing evidence suggests that the mechanisms underpinning the beneficial effects of such cells in muscle tissue reside in their ability to secrete bioactive molecules (secretome) with multiple actions. Hence, we examined the effects of MSC secretome as conditioned medium (MSC-CM) on ex vivo murine extensor digitorum longus muscle injured by forced eccentric contraction (EC). By combining morphological (light and confocal laser scanning microscopies) and electrophysiological analyses we demonstrated the capability of MSC-CM to attenuate EC-induced tissue structural damages and sarcolemnic functional properties’ modifications. MSC-CM was effective in protecting myofibers from apoptosis, as suggested by a reduced expression of pro-apoptotic markers, cytochrome c and activated caspase-3, along with an increase in the expression of pro-survival AKT factor. Notably, MSC-CM also reduced the EC-induced tissue redistribution and extension of telocytes/CD34⁺ stromal cells, distinctive cells proposed to play a “nursing” role for the muscle resident myogenic satellite cells (SCs), regarded as the main players of regeneration. Moreover, it affected SC functionality likely contributing to replenishment of the SC reservoir. This study provides the necessary groundwork for further investigation of the effects of MSC secretome in the setting of skeletal muscle injury and regenerative medicine.     

Effect of HCl pre-treatment on corrosion resistance of cerium-based conversion coatings on magnesium and magnesium alloys

The corrosion protection afforded by a cerium conversion coating, formed by immersion in a solution containing rare earth salt and hydrogen peroxide, on pure magnesium and two magnesium alloys, AZ91 and AM50, has been studied. The effect of HCl pre-treatments on the morphology and on the corrosion resistance of the cerium conversion layer was investigated. A thicker and more homogeneous distribution of the conversion coating was obtained when the sample surface was pre-treated with acid. Higher amounts of cerium on the surface of the pre-treated samples were detected. The cerium conversion coating increased the corrosion resistance of the alloys because it ennobled the corrosion potential and decreased both the anodic and cathodic current. The acid pre-treatment further increased the corrosion resistance of the coated alloys. After five days of immersion in chloride environment the untreated samples showed localized corrosion while the chemical conversion coated samples appeared unaffected.     

Effect of processing on sphingolipid content in soybean products

Soybeans are believed to be a rich source of sphingolipids, a class of polar lipids that has received attention for their possible cancer-inhibiting activities. The effect of processing on the sphingolipid content of various soybean products has not been determined. Glucosylceramide (GlcCer), the major sphingolipid type in soybeans, was measured in several processed soybean products to illustrate which product(s) GlcCer is partitioned into during processing and where it is lost. Whole soybeans were processed into full-fat flakes, from which crude oil was extracted. Crude oil was refined by conventional methods, and defatted soy flakes were further processed into alcohol-washed and acid-washed soy protein concentrates (SPC) and soy protein isolates (SPI) by laboratory-scale methods that simulated industrial practices. GlcCer was isolated from the samples by solvent extraction, solvent partition, and TLC and was quantified by HPLC. GlcCer remained mostly within the defatted soy flakes (91%) rather than in the oil (9%) after oil extraction. Only 52, 42, and 26% of GlcCer from defatted soy flakes was recovered in the acid-washed SPC, alcohol-washed SPC, and SPI products, respectively. All protein products had a similar GlcCer concentration of about 281 nmol/g (dry wt basis). The minor quantity of GlcCer in the crude oil was almost completely removed by water degumming.     

Synthesis and properties of aminopropyl nucleic acids

Oligonucleotides that contain up to three aminopropyl nucleoside analogues have been synthesized. Dimers of aminopropyl adenine and thymidine were prepared and used as building blocks by applying phosphoramidite chemistry. Both R and S isomers of the aminopropyl nucleosides were used. This incorporation led to a reduction of thermal stability of double-stranded DNA. Furthermore, the (R)-adenine analogue, which yielded (S)-APNA, can be considered as a candidate for universal base pairing.     

Suppression of carbon monoxide formation in oxide-coated TZM molybdenum X-ray rotating anodes

Higher performance designs for rotating anode X-ray tubes have increased the average rotating anode temperature from below 1100 °C to well above 1300 °C. This temperature increase has accelerated the formation of carbon monoxide by reaction of carbon from the alloy substrate with oxygen from the emissive coating. The dominant carbon source is thought to be Mo₂C grain boundary precipitates in the TZM molybdenum alloy substrate. The dominant oxygen source is thought to be TiO in the emissive coating. Placement of a monocarbide-forming reactive layer between the alloy substrate and the emissive coating has been demonstrated to lower the thermodynamic activity of the carbon source and dramatically reduce the rate of formation of carbon monoxide.     

Understanding student participation in undergraduate course communities: A case study

Participation is the cornerstone of any community. Promoting, understanding and properly managing it allows not only keeping the community sustainable, but also providing personalized services to its members and managers. This article presents a case study in which student participation in a course community was motivated using two different extrinsic mechanisms, and mediated by a software platform. The results were compared with a baseline community of the same course, in which participation was not motivated by external means. The analysis of these results indicates that managing a partially virtual course community requires the introduction of monitoring services, community managers and extrinsic mechanisms to motivate participation. These findings allow community managers to improve their capability for promoting participation and keeping the community sustainable. The findings also raise several implications that should be considered in the design of software supporting this kind of community, when managing the participation of its members.     

A laboratory assessment of the impact of brewery wastewater discharge on sulfide and methane production in a sewer

The impact of brewery wastewater discharge on sulfide and methane production in a sewer was assessed. Experiments were carried out on laboratory scale sewer reactors consisting of both an experimental and a control reactor. The control reactor was intermittently fed with real fresh sewage while the experimental reactor was fed with a mixture of brewery and domestic wastewater at two different proportions (10 and 25% v/v). 10% v/v discharge of brewery wastewater increased the H2S and CH4 production rates in the sewer reactor by 40% and 30%, respectively. When the brewery wastewater fraction was increased to 25% v/v, the H2S production rate of the experimental reactor decreased to the level of the control reactor. In contrast, the CH4 production rate maintained at a level that was 30% higher than that in the control reactor. These results indicate that the discharge of brewery wastewater into sewers can give negative impacts in relation to odour and corrosion management of the systems and will increase the greenhouse gas emissions from sewers. The study also reveals that the impact of trade waste on the biological reactions in sewers is complex, and requires careful experimental assessment in each case.     

Texture and shape information fusion for facial expression and facial action unit recognition

A novel method based on fusion of texture and shape information is proposed for facial expression and Facial Action Unit (FAU) recognition from video sequences. Regarding facial expression recognition, a subspace method based on Discriminant Non-negative Matrix Factorization (DNMF) is applied to the images, thus extracting the texture information. In order to extract the shape information, the system firstly extracts the deformed Candide facial grid that corresponds to the facial expression depicted in the video sequence. A Support Vector Machine (SVM) system designed on an Euclidean space, defined over a novel metric between grids, is used for the classification of the shape information. Regarding FAU recognition, the texture extraction method (DNMF) is applied on the differences images of the video sequence, calculated taking under consideration the neutral and the expressive frame. An SVM system is used for FAU classification from the shape information. This time, the shape information consists of the grid node coordinate displacements between the neutral and the expressed facial expression frame. The fusion of texture and shape information is performed using various approaches, among which are SVMs and Median Radial Basis Functions (MRBFs), in order to detect the facial expression and the set of present FAUs. The accuracy achieved using the Cohn–Kanade database is 92.3% when recognizing the seven basic facial expressions (anger, disgust, fear, happiness, sadness, surprise and neutral), and 92.1% when recognizing the 17 FAUs that are responsible for facial expression development.