Cellular Integrin α5β1 and Exosomal ADAM17 Mediate the Binding and Uptake of Exosomes Produced by Colorectal Carcinoma Cells

Approximately 25% of colorectal cancer (CRC) patients develop peritoneal metastasis, a condition associated with a bleak prognosis. The CRC peritoneal dissemination cascade involves the shedding of cancer cells from the primary tumor, their transport through the peritoneal cavity, their adhesion to the peritoneal mesothelial cells (PMCs) that line all peritoneal organs, and invasion of cancer cells through this mesothelial cell barrier and underlying stroma to establish new metastatic foci. Exosomes produced by cancer cells have been shown to influence many processes related to cancer progression and metastasis. In epithelial ovarian cancer these extracellular vesicles (EVs) have been shown to favor different steps of the peritoneal dissemination cascade by changing the functional phenotype of cancer cells and PMCs. Little is currently known, however, about the roles played by exosomes in the pathogenesis and peritoneal metastasis cascade of CRC and especially about the molecules that mediate their interaction and uptake by target PMCs and tumor cells. We isolated exosomes by size−exclusion chromatography from CRC cells and performed cell-adhesion assays to immobilized exosomes in the presence of blocking antibodies against surface proteins and measured the uptake of fluorescently-labelled exosomes. We report here that the interaction between integrin α5β1 on CRC cells (and PMCs) and its ligand ADAM17 on exosomes mediated the binding and uptake of CRC-derived exosomes. Furthermore, this process was negatively regulated by the expression of tetraspanin CD9 on exosomes.     

Inherited Retinal Diseases Due to RPE65 Variants: From Genetic Diagnostic Management to Therapy

Inherited retinal diseases (IRDs) are a heterogeneous group of conditions that include retinitis pigmentosa (RP) and Leber congenital amaurosis (LCA) and early-onset severe retinal dystrophy (EO[S]RD), which differ in severity and age of onset. IRDs are caused by mutations in >250 genes. Variants in the RPE65 gene account for 0.6–6% of RP and 3–16% of LCA/EORD cases. Voretigene neparvovec is a gene therapy approved for the treatment of patients with an autosomal recessive retinal dystrophy due to confirmed biallelic RPE65 variants (RPE65-IRDs). Therefore, the accurate molecular diagnosis of RPE65-IRDs is crucial to identify ‘actionable’ genotypes—i.e., genotypes that may benefit from the treatment—and is an integral part of patient management. To date, hundreds of RPE65 variants have been identified, some of which are classified as pathogenic or likely pathogenic, while the significance of others is yet to be established. In this review, we provide an overview of the genetic diagnostic workup needed to select patients that could be eligible for voretigene neparvovec treatment. Careful clinical characterization of patients by multidisciplinary teams of experts, combined with the availability of next-generation sequencing approaches, can accelerate patients’ access to available therapeutic options.     

Low prevalence of Listeria monocytogenes in cull sows and pork

The goal of this study was to determine the prevalence of Listeria monocytogenes in sows slaughtered at a single Midwestern plant on two occasions (trial 1, n = 179 sows; trial 2, n = 160 sows). Fecal samples collected antemortem (trial 1) as well as animal tissues, and carcass swabs collected at the abattoir (trials 1 and 2) were analyzed. Eight isolates of L. monocytogenes were recovered from five samples that represented 0.18% of the total samples (n = 2,775). In trial 1, L. monocytogenes was detected in a tonsil sample (0.6%; 1 positive of 181 tonsils), in a carcass (0.6%; 1 positive of 179 carcasses), which was sampled prior to the organic rinse, and in two chopped meat block samples (1.2%; 2 positive of 165 samples). In trial 2, L. monocytogenes was only detected in a single chopped meat block sample (0.15%; 1 positive of 688 total samples). These data indicate the low prevalence of L. monocytogenes in the cull sow.     

Particle shape control using pulse electrodeposition: Methanol oxidation as a probe reaction on Pt dendrites and cubes

Shape controlled Pt particles were synthesized onto tungsten monocarbide (WC) substrates using a pulse electrodeposition method. The particle shape was strongly influenced by the deposition potential, with cubic particles formed using 0.14 V vs. NHE (normal hydrogen electrode) and dendritic particles formed at 0.04 V vs. NHE. The crystalline orientation and active surface area of the Pt particles were estimated using Cu stripping voltammetry. Finally, cyclic voltammetry and chronoamperometry were used to determine the methanol electrooxidation activity, which revealed that the dendritic Pt showed much higher electrochemical activity than the cubic particles. These results demonstrated the possibility of more effectively utilizing Pt electrocatalysts by controlling the shape of Pt particles.     

Oxide/polymer interfaces for hybrid and organic solar cells: Anatase vs. Rutile TiO2

In this work, we study the effect of the transparent conducting oxide (TCO) and the polymer applied (MEH-PPV or P3HT) on the photovoltaic properties of TCO/TiO₂/polymer/Ag bi-layer solar cells. The solar cells were analyzed under inert atmosphere conditions resembling an encapsulated or sealed device. We demonstrate that the substrate applied, ITO or FTO, modifies the crystalline structure of the TiO₂: on an ITO substrate, TiO₂ is present in its anatase phase, on an FTO, the rutile phase predominates. Devices fabricated on an FTO, where the rutile phase is present, show better stability under inert atmospheres than devices fabricated on an ITO, anatase phase. With respect to the polymer, devices based on MEH-PPV show higher Voc (as high as 1 V), while the application of P3HT results in lower Voc, but higher J_sc and longer device stability. These observations have been associated to (a), the crystalline structure of TiO₂ and (b) to the form the polymer is bonded to the TiO₂ surface. In-situ IPCE analyses of P3HT-based solar cells show a red shift on the peak corresponding to TiO₂, which is not present on the MEH-PPV-based solar cells. The latter suggest that P3HT can be linked to the TiO₂ though the S-end atom, which results in devices with lower Voc. All these observations are also valid for devices, where the bare TiO₂ is replaced by an Nb-TiO₂. The application of an Nb-TiO₂ with rutile structure in these polymer/oxide solar cells is the reason for their higher stability under inert atmospheres. We conclude that the application of TiO₂ in its rutile phase is beneficial for long-term stability devices. Moreover there is an interplay between low Voc and J_sc in devices applying P3HT, since power conversion efficiency can be partially canceled by their lower Voc in comparison with MEH-PPV. These findings are important for polymer/oxide solar cells, but also for organic solar cells, where a layer of semiconductor oxides are in direct contact with a polymer, like in an inverted or tandem organic solar cells.     

Environmental exposure assessment of fluoroquinolone antibacterial agents from sewage to soil

The behavior of fluoroquinolone antibacterial agents (FQs) during mechanical-biological wastewater treatment was studied by mass flow analysis. In addition, the fate of FQs in agricultural soils after sludge application was investigated. Concentrations of FQs in filtered wastewater (raw sewage, primary, secondary, and tertiary effluents) were determined using solid-phase extraction with mixed phase cation exchange disk cartridges and reversed-phase liquid chromatography with fluorescence detection. FQs in suspended solids, sewage sludge (raw, excess, and anaerobically digested sludge), and sludge-treated soils were determined as described for the aqueous samples but preceded by accelerated solvent extraction. Wastewater treatment resulted in a reduction of the FQ mass flow of 88-92%, mainly due to sorption on sewage sludge. A sludge-wastewater partition coefficient (log Kd approximately 4) was calculated in the activated sludge reactors with a hydraulic residence time of about 8 h. No significant removal of FQs occurred under methanogenic conditions of the sludge digesters. These results suggest sewage sludge as the main reservoir of FQ residues and outline the importance of sludge management strategies to determine whether most of the human-excreted FQs enter the environment. Field experiments of sludge-application to agricultural land confirmed the long-term persistence of trace amounts of FQs in sludge-treated soils and indicated a limited mobility of FQs into the subsoil.     

Chemical composition,nutritional value and antioxidant properties of Allium caepa L. Var. tropeana (red onion) seeds

Chemical analysis of Allium caepa L. var Tropeana (red onion) seeds showed high amounts of oil (20.4%), fibre (22.4%), crude protein (24.8%), calcium (175.0 mg/100 g), potassium (1010 mg/100 g), low amounts of sodium (11.2 mg/100 g) and six cysteine derivatives, of which the S-propylmercapto-cysteine has never been reported in onion before. The antioxidant capacity of seed extracts containing cysteine derivatives (SECCD), before and after boiling the seeds, and of cooking water extracts containing cysteine derivatives (CWECCD), was also evaluated, by the ferric reducing antioxidant power (FRAP) and 1,1-diphenyl-2-picrilhydrazyl (DPPH) assays. The extracts showed discrete antioxidant capacity which increased after boiling, although cooking methods caused significant losses of the cysteine derivatives in water.     

Bacteriological quality of raw oysters in Trinidad and the attitudes, knowledge and perceptions of the public about its consumption

In Trinidad and Tobago, raw oyster cocktails are a delicacy, but they are generally believed to be responsible for illness in the consumer. The microbial loads of raw oysters, condiments/spices and ready-to-consume oyster cocktails were determined in four sampling areas. Questionnaires were also administered to 72 oyster vendors to determine practices that affect the bacteriological quality of the products. Three hundred members of the public were interviewed to determine the attitudes, knowledge and perceptions of the public about raw oyster consumption. The mean total aerobic plate count (TAPC) per g of 50 samples each, of raw oysters, condiments/spices and ready-to-consume raw oyster cocktails averaged from 1.0 x 10(7)+/-4.3 x 10(7) to 1.4 x 10(8)+/-6.4 x 10(8), 2.0 x 10(5)+/-1.0 x 10(6) to 2.0 x 10(7)+/-1.4 x 10(8), and 4.3 x 10(5)+/-1.0 x 10(6) to 2.2 x 10(6)+/-1.0 x 10(7), respectively. The difference for each product among the four areas was statistically significant (P < or = 0.05; chi2). Using a recommended maximum standard of TAPC per g of 5.0 x 10(5), 115 (57.5%), 27 (13.5%) and 51 (25.5%) of 200 samples each, of raw oysters, condiments/spices and oyster cocktails, respectively, were considered unfit for human consumption. Amongst vendor practices, source of oyster harvest and length of time between separation of oyster meat from shell and sale, significantly affected the mean TAPC per g and the prevalence of unfit oyster cocktail samples. Consumption of raw oyster cocktails was more prevalent amongst males (73.6%) than females (26.4%) (P < 0.002), East Indians (63.2%) as compared with other respondents (36.8%) (P < 0.001), individuals < or = 40 years old (82.1%) than in individuals > 40 years of age (17.9%) (P < 0.01), and in individuals who were aware that raw oysters are considered to be a sexual enhancer (86.8%) as compared to those who did not have this perception (11.3%) (P < 0.03). Fear of falling ill prevented 44 (37.9%) of 116 non-consumers from eating oyster cocktails, while 13 of 106 consumers (12.3%) reported having experienced an oyster-borne illness. The rather high prevalence of raw oyster cocktails found to be unfit for human consumption, coupled with the perceptions and attitudes of the consumers about the product, pose a significant health risk to the public.     

Polysaccharidzus?tze in eiwei?reichen Lebensmitteln

Zusammenfassung Aus der Literatur bekannte Verfahren zur Isolierung von Polysacchariden aus eiweißreichen Lebensmitteln, insbesondere Frischkäsen, erwiesen sich im praktisch interessanten Bereich (0,01–0,2% Bindemittelzusatz) als wenig brauchbar. Stattdessen wird folgende Probenaufbereitung vorgeschlagen: 1. Entfettung der Proben mit einem Gemisch von Äthanol, Diäthyläther und Petroläther; 2. vollständige Hydrolyse der Proteine mit Pepsin, Pronase E und einem Gemisch von Aminopeptidase M und Prolidase; 3. Abtrennung der Polysaccharide von den niedermolekularen Substanzen und von Enzymresten durch Gelfiltration an Biogel P-2; 4. Konzentrierung der polysaccharidhaltigen Fraktion durch Gefriertrocknung, Ultrafiltration oder Eindampfen.- Das Verfahren wurde am Beispiel von Speisequark und Doppelrahmfrischkäse erprobt, ist im Prinzip aber verallgemeinerungsfähig.

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Polysaccharide additives in foods rich in proteinsI. Sample preparation for estimation, especially in fresh cheese

Summary Published procedures for the isolation of polysaccharides from foods rich in proteins, especially fresh cheese, proved not very useful for the concentration in question (0,01–0.2%). We, instead, propose the following sample preparation: 1. Removal of lipids by an ethanol - ethyl ether - petrol ether mixture; 2. complete hydrolysis of proteins with pepsin, pronase E and a mixture of Amnopeptidase M and prolidase; 3. separation of the polysaccharides from the low molecular substances and from enzyme residues by gel filtration on Biogel P-2; 4. concentration of the polysaccharides containing fraction by freeze drying, ultrafiltration or evaporation. - The method was tested with quarg and double cream fresh cheese, it can, however, be adopted to use in other products.

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Deutsches Wollforschungsinstitut, Aachen.     

Spatial Input for Temporal Navigation in Scientific Visualizations

Scientific-visualization tools can make time-varying simulations easier to understand. The growing efficiency of today's high-performance computers enables simulation of physical phenomena with a high temporal resolution. Consequently, visualization systems require efficient navigation in the temporal dimension. This 3D user interface employs direct-manipulation metaphors for temporal navigation in scientific visualizations. By interacting with objects using their 3D trajectory, users can navigate in time by specifying spatial inputs. This article is part of a special issue on 3D user interfaces.