Identification of fungemia agents using the polymerase chain reaction and restriction fragment length polymorphism analysis

Prompt and specific identification of fungemia agents is important in order to define clinical treatment. However, in most cases conventional culture identification can be considered to be time-consuming and not without errors. The aim of the present study was to identify the following fungemia agents: Candida albicans, Candida parapsilosis, Candida tropicalis, Candida glabrata, Cryptococcus neoformans, Cryptococcus gattii, and Histoplasma capsulatum using the polymerase chain reaction and restriction fragment length polymorphism analysis (PCR/RFLP). More specifically: a) to evaluate 3 different amplification regions, b) to investigate 3 different restriction enzymes, and c) to use the best PCR/RFLP procedure to indentify 60 fungemia agents from a culture collection. All 3 pairs of primers (ITS1/ITS4, NL4/ITS5 and Primer1/Primer2) were able to amplify DNA from the reference strains. However, the size of these PCR products did not permit the identification of all the species studied. Three restriction enzymes were used to digest the PCR products: HaeIII, Ddel and Bfal. Among the combinations of pairs of primers and restriction enzymes, only one (primer pair NL4/ITS5 and restriction enzyme Ddel) produced a specific RFLP pattern for each microorganism studied. Sixty cultures of fungemia agents (selected from the culture collection of Fundação de Medicina Tropical do Amazonas - FMTAM) were correctly identified by PCR/RFLP using the prime pair NL4/ITS5 and Ddel. We conclude that the method proved to be both simple and reproducible, and may offer potential advantages over phenotyping methods.

Evaluation of anti-Wnt/β-catenin signaling agents by pGL4-TOP transfected stable cells with a luciferase reporter system

Refractory and relapsed leukemia is a major problem during cancer therapy, which is due to the aberrant activation of Wnt/β-catenin signaling pathway. Activation of this pathway is promoted by wingless (Wnt) proteins and induces co-activator β-catenin binding to lymphoid enhancer factor (LEF)/T-cell factor protein (TCF). To provide a convenient system for the screening of anti-Wnt/β-catenin agents, we designed a bi-functional pGL4-TOP reporter plasmid that contained 3X β-catenin/LEF/TCF binding sites and a selectable marker. After transfection and hygromycin B selection, HEK 293-TOP and Jurkat-TOP stable clones were established. The luciferase activity in the stable clone was enhanced by the recombinant Wnt-3A (rWnt-3A; 100-400 ng/mL) and GSK3β inhibitor (2’Z,3’E)-6-bromoindirubin-3’-oxime (BIO; 5 µM) but was inhibited by aspirin (5 mM). Using this reporter model, we found that norcantharidin (NCTD; 100 µM) reduced 80% of rWnt-3A-induced luciferase activity. Furthermore, 50 µM NCTD inhibited 38% of BIO-induced luciferase activity in Jurkat-TOP stable cells. Employing ³H-thymidine uptake assay and Western blot analysis, we confirmed that NCTD (50 µM) significantly inhibited proliferation of Jurkat cells by 64%, which are the dominant β-catenin signaling cells and decreased β-catenin protein in a concentration-dependent manner. Thus, we established a stable HEK 293-TOP clone and successfully used it to identify the Wnt/β-catenin signaling inhibitor NCTD.

Prevention of enzymatic browning of yacon flour by the combined use of anti-browning agents and the study of its chemical composition

Yacon roots present functional properties because of the high levels of fructooligosaccharides (FOS), which are considered as prebiotic fibers. In addition, yacon roots are rich in phenolic compounds. During the processing of yacon, the freshly cut surface undergoes rapid enzymatic browning. Control of enzymatic browning during processing is very important to preserve the appearance of yacon flour. In this study, it was evaluated the combined effect of anti-browning agents (ascorbic acid, citric acid and L-cysteine) on the inhibition of enzymatic browning of yacon, using Response Surface Methodology. The yacon pre-treated with anti-browning agents in concentrations of 15.0 mM for ascorbic acid, 7.5 mM for citric acid and 10.0 mM for L-cysteine was used for the processing of flour. Yacon flour presented an attractive color and good sensory properties, without residual aroma. The contents of FOS and phenolic compounds obtained in yacon flour were 28.60 g.100 g- 1 and 1.35 g.100 g- 1. Yacon flour can be considered as a potential functional food, especially due to high levels of FOS, which allows for its use in formulation of various foods.

Marks of Fire, Value and Faith. Swords with Ferrous Inlays in Finland during the Late Iron Age (ca. 700–1200 AD)

This study explores swords with ferrous inlays found in Finland and dating from the late Iron Age, ca. 700–1200 AD. These swords reflect profound changes not only in styles and fashion but also in the technology of hilts and blades. This study explores how many of these kinds of swords are known from Finland, how they were made and where, what their status was in Late Iron Age Finland, and where the Finnish finds stand in accordance with other areas of Europe. The various methods included measuring of the finds and statistics. The main method of revealing the inlaid marks was radiography due to its non-destructive nature. In cases where inlays were visible without radiography, their details were inspected via microscopy. To study the materials and manufacture of inlaid swords, a sample of them was metallographically analysed to determine the forging technologies and nature of used materials. Furthermore, the manufacture was also studied with experimental approaches. As a result, a catalogue of 151 swords with ferrous inlays was created. This number is relatively high compared with other European countries, although systematic studies have been conducted in only some countries. The inlaid motifs were classified into five distinct categories to help the classification. To summarize, almost every documented inlaid sword was unique in some respect including measurements, inlaid motifs and materials of blades and inlays. Technological variation was also present, some blades being poorer and some of higher quality in spite of the inlaid motifs. Misspelt inscriptions as well as letter-like marks were common in Finland and also in Scandinavia. Furthermore, the provenance of iron and steel used in some blades hints at Scandinavian ores. The above observations, along with the experimental results indicating the existence of multiple alternative techniques of inlaying, suggest that these swords were manufactured locally in Scandinavia, most likely in imitation of Continental European models. Inlaid swords were valued partly for their assumed functionality in combat, as evidenced by damage on some examined blades, or they were valued for their inlays, which could have had fashionable or symbolical meanings bound to local beliefs.

Cours de construction ; 1re-2e parties, 1-2. Cours de construction destiné aux conducteurs et employés des Ponts et chaussées, agents-voyers, architectes, garde-mines, employés des compagnies de chemins de fer, entrepreneurs, maîtres ouvriers (charpentiers, menuisiers, serruriers, maçons, tailleurs de pierre, etc.), et à toutes les personnes s'occupant de travaux à un titre quelconque. Matériaux de construction et leur emploi / par Gustave Oslet,...

Collection : Encyclopédie théorique et pratique des connaissances civiles et militaires ; partie 1, livre 6

Cours de construction ; 1re-2e parties, 1-2. Cours de construction destiné aux conducteurs et employés des Ponts et chaussées, agents-voyers, architectes, garde-mines, employés des compagnies de chemins de fer, entrepreneurs, maîtres ouvriers (charpentiers, menuisiers, serruriers, maçons, tailleurs de pierre, etc.), et à toutes les personnes s'occupant de travaux à un titre quelconque. Traité de géodésie / par Gustave Oslet,...

Collection : Encyclopédie théorique et pratique des connaissances civiles et militaires ; partie 1, livre 6

Bulletin consulaire français : recueil des rapports commerciaux adressés au Ministère des affaires étrangères par les agents diplomatiques de France à l'étranger

1887/01 (VOL13)-1887/06.

Bulletin consulaire français : recueil des rapports commerciaux adressés au Ministère des affaires étrangères par les agents diplomatiques de France à l'étranger

1881 (A1881).

Bulletin consulaire français : recueil des rapports commerciaux adressés au Ministère des affaires étrangères par les agents diplomatiques de France à l'étranger

1887/07 (VOL13)-1887/12.

Bulletin consulaire français : recueil des rapports commerciaux adressés au Ministère des affaires étrangères par les agents diplomatiques de France à l'étranger

1885 (A1885).

Bulletin consulaire français : recueil des rapports commerciaux adressés au Ministère des affaires étrangères par les agents diplomatiques de France à l'étranger

1877 (A1877).

Bulletin consulaire français : recueil des rapports commerciaux adressés au Ministère des affaires étrangères par les agents diplomatiques de France à l'étranger

1883 (A1883).