992 resultados para Enriched genomic library


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Iron is an essential element for nearly all living organisms, and its deficiency is the most common form of malnutrition in the world. The organic forms of trace elements are considered more bioavailable than the inorganic forms. Although Saccharomyces cerevisiae can enrich metal elements and convert inorganic iron to organic species, its tolerability and transforming capacity are limited. The aim of this study was to screen higher biomass and other iron-enriched fungi strains besides Saccharomyces cerevisiae from the natural environment. A PDA medium containing 800 μg/mL iron was used for initial screening. Fifty strains that tolerated high iron concentration were isolated from the natural environment, and only one strain, No.BY1109, grew well at Fe (II) concentration of 10,000μg/ml. According to morphological characterization, 18S rDNA sequence analysis, and biophysical and biochemical characterization, the strain No.BY1109 was identified as Rhodotorula. The iron content of No.BY1109 (10 mg Fe/g dry cell) was determined using atomic absorption spectrometry. The results of distribution of iron in the cells showed that iron ion was mainly chelated in the cell walls and vacuoles. The bioavailability in rats confirmed that strain No.BY1109 had higher absorption efficiency than that of ferrous sulfate after single dose oral administration. The present study introduces new iron supplements, and it is a basis for finding new iron supplements from natural environment.

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AbstractEffects of different β-glucan concentrations in maize flour on the properties of frozen maize tortillas were evaluated. Masa (dough), pre-cooked (PTs), frozen (FTs), thawed (TTs), and cooked tortillas (CTs) were made and analyzed. Moisture content of masa and tortillas significantly decreased as β-glucan concentration increased; however, the water absorption capacity (WAC), ice melting enthalpy, and frozen water in FTs increased. Texture and color of the masa, PTs, and CTs as well as sensory analysis showed differences only between tortillas with 0% and 4% β-glucans. β-glucans did not affect the texture of CTs. Soluble fiber increased by over threefold and fivefold in tortillas with 2% and 4% β-glucans, respectively, than in those without β-glucans. This result was consistent with the observed structural changes in tortillas, showing an increase in high-fiber aggregates with increasing β-glucan concentration. Tortillas with 2% β-glucans showed acceptable physicochemical, functional, and sensory properties, but over three times the soluble fiber. Therefore, it is possible to obtain frozen tortillas with high fiber content and increase their shelf life for subsequent cooking while maintaining good properties.

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Abstract The present work aimed at studying the effect of different drying methods applied to tilapia byproducts (heads, viscera and carcasses) fed with flaxseed, verifying the contents of omega-3 fatty acids. Two diets were given to the tilapia: a control and a flaxseed formulation, over the course of 60 days. After this period, they were slaughtered and their byproducts (heads, viscera and carcasses) were collected. These fish parts were analyzed in natura, lyophilized and oven dried. Byproducts from tilapia fed with flaxseed presented docosapentaenoic, eicopentaenoic and docosahexanoic fatty acids as a result of the enzymatic metabolism of the fish. The byproducts from the oven drying process had lower levels of polyunsaturated fatty acids. In the multivariate analysis, the byproducts from fish fed with flaxseed had a greater composition of fatty acids. The addition of flaxseed in fish diets, as well as the utilization of their byproducts, may become a good business strategy. Additionally, the byproducts may be dried to facilitate transport and storage.

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Teema: Kansainvälisyys.

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Cancer affects more than 20 million people each year and this rate is increasing globally. The Ras/MAPK-pathway is one of the best-studied cancer signaling pathways. Ras proteins are mutated in almost 20% of all human cancers and despite numerous efforts, no effective therapy that specifically targets Ras is available to date. It is now well established that Ras proteins laterally segregate on the plasma membrane into transient nanoscale signaling complexes called nanoclusters. These Ras nanoclusters are essential for the high-fidelity signal transmission. Disruption of nanoclustering leads to reduction in Ras activity and signaling, therefore targeting nanoclusters opens up important new therapeutic possibilities in cancer. This work describes three different studies exploring the idea of membrane protein nanoclusters as novel anti-cancer drug targets. It is focused on the design and implementation of a simple, cell-based Förster Resonance Energy Transfer (FRET)-biosensor screening platform to identify compounds that affect Ras membrane organization and nanoclustering. Chemical libraries from different sources were tested and a number of potential hit molecules were validated on full-length oncogenic proteins using a combination of imaging, biochemical and transformation assays. In the first study, a small chemical library was screened using H-ras derived FRET-biosensors. Surprisingly from this screen, commonly used protein synthesis inhibitors (PSIs) were found to specifically increase H-ras nanoclustering and downstream signalling in a H-ras dependent manner. Using a representative PSI, increase in H-ras activity was shown to induce cancer stem cell (CSC)-enriched mammosphere formation and tumor growth of breast cancer cells. Moreover, PSIs do not increase K-ras nanoclustering, making this screening approach suitable for identifying Ras isoform-specific inhibitors. In the second study, a nanoncluster-directed screen using both H- and K-ras derived FRET biosensors identified CSC inhibitor salinomycin to specifically inhibit K-ras nanocluster organization and downstream signaling. A K-ras nanoclusteringassociated gene signature was established that predicts the drug sensitivity of cancer cells to CSC inhibitors. Interestingly, almost 8% of patient tumor samples in the The Cancer Genome Atlas (TCGA) database had the above gene signature and were associated with a significantly higher mortality. From this mechanistic insight, an additional microbial metabolite screen on H- and K-ras biosensors identified ophiobolin A and conglobatin A to specifically affect K-ras nanoclustering and to act as potential breast CSC inhibitors. In the third study, the Ras FRET-biosensor principle was used to investigate membrane anchorage and nanoclustering of myristoylated proteins such as heterotrimeric G-proteins, Yes- and Src-kinases. Furthermore, Yes-biosensor was validated to be a suitable platform for performing chemical and genetic screens to identify myristoylation inhibitors. The results of this thesis demonstrate the potential of the Ras-derived FRETbiosensor platform to differentiate and identify Ras-isoform specfic inhibitors. The results also highlight that most of the inhibitors identified predominantly perturb Ras subcellular distribution and membrane organization through some novel and yet unknown mechanisms. The results give new insights into the role of Ras nanoclusters as promising new molecular targets in cancer and in stem cells.

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Pictured here from left to right - Back Row: John Burtniak, Cataloguing. Nick Krenton, Head Cataloguing. Front Row: Sylvia Osterbind, Reference. Arthur Vespry, Chief Librarian. Mara Karnupe, Technical Services. Dianna Kertland, Circulation.

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View of the interior of the original library.

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Catharanthus roseus is the sole biological source of the medicinal compounds vinblastine and vincristine. These chemotherapeutic compounds are produced in the aerial organs of the plant, however they accumulate in small amounts constituting only about 0.0002% of the fresh weight of the leaf. Their limited biological supply and high economical value makes its biosynthesis important to study. Vinblastine and vincristine are dimeric monoterpene indole alkaloids, which consists of two monomers vindoline and catharanthine. The monoterpene indole alkaloids (MIA's) contain a monoterpene moiety which is derived from the iridoid secologanin and an indole moiety tryptamine derived from the amino acid tryptophan. The biosynthesis of the monoterpene indole alkaloids has been localized to at least three cell types namely, the epidermis, the laticifer and the internal phloem assisted parenchyma. Carborundum abrasion (CA) technique was developed to selectively harvest epidermis enriched plant material. This technique can be used to harvest metabolites, protein or RNA. Sequencing of an expressed sequence tagged (EST) library from epidermis enriched mRNA demonstrated that this cell type is active in synthesizing a variety of secondary metabolites namely, flavonoids, lipids, triterpenes and monoterpene indole alkaloids. Virtually all of the known genes involved in monterpene indole alkaloid biosynthesis were sequenced from this library.This EST library is a source for many candidate genes involved in MIA biosynthesis. A contig derived from 12 EST's had high similarity (E'^') to a salicylic acid methyltransferase. Cloning and functional characterization of this gene revealed that it was the carboxyl methyltransferase imethyltransferase (LAMT). In planta characterization of LAMT revealed that it has a 10- fold enrichment in the leaf epidermis as compared to the whole leaf specific activity. Characterization of the recombinant enzyme revealed that vLAMT has a narrow substate specificity as it only accepts loganic acid (100%) and secologanic acid (10%) as substrates. rLAMT has a high Km value for its substrate loganic acid (14.76 mM) and shows strong product inhibition for loganin (Kj 215 |iM). The strong product inhibition and low affinity for its substrate may suggest why the iridoid moiety is the limiting factor in monoterpene indole alkaloid biosynthesis. Metabolite profiling of C. roseus organs shows that secologanin accumulates within these organs and constitutues 0.07- 0.45% of the fresh weight; however loganin does not accumulate within these organs suggesting that the product inhibition of loganin with LAMT is not physiologically relevant. The limiting factor to iridoid and MIA biosynthesis seems to be related to the spatial separation of secologanin and the MIA pathway, although secologanin is synthesized in the epidermis, only 2-5% of the total secologanin is found in the epidermis while the remaining secologanin is found within the leaf body inaccessable to alkaloid biosynthesis. These studies emphasize the biochemical specialization of the epidermis for the production of secondary metabolites. The epidermal cells synthesize metabolites that are sequestered within the plant and metabolites that are secreted to the leaf surface. The secreted metabolites comprise the epidermome, a layer separating the plant from its environment.

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A history of Niagara Falls, including the Programme for the Conference.

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Clara Irvin Chapman's last visit to the Chapman campus was in March, 1967 for the opening of the Thurmond Clarke Memorial Library. She is in the Heritage Room by a portrait of her late husband, Charles Clarke Chapman, for whom the college is named.

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Clara Irvin Chapman's last visit to the Chapman campus was in March, 1967 for the opening of the Thurmond Clarke Memorial Library. She is in the Heritage Room by a portrait of her late husband, Charles Clarke Chapman, for whom the college is named.

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Clara Irvin Chapman's last visit to the Chapman campus was in March, 1967 for the opening of the Thurmond Clarke Memorial Library. She is in the Heritage Room of the library.

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Clara Irvin Chapman's last visit to the Chapman campus was in March, 1967 for the opening of the Thurmond Clarke Memorial Library. She is in the Heritage Room by a portrait of her late husband, Charles Clarke Chapman, for whom the college is named.

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