Bacterial Community Composition in Brazilian Anthrosols and Adjacent Soils Characterized Using Culturing and Molecular Identification

Microbial community composition was examined in two soil types, Anthrosols and adjacent soils, sampled from three locations in the Brazilian Amazon. The Anthrosols, also known as Amazonian dark earths, are highly fertile soils that are a legacy of pre-Columbian settlement. Both Anthrosols and adjacent soils are derived from the same parent material and subject to the same environmental conditions, including rainfall and temperature; however, the Anthrosols contain high levels of charcoal-like black carbon from which they derive their dark color. The Anthrosols typically have higher cation exchange capacity, higher pH, and higher phosphorus and calcium contents. We used culture media prepared from soil extracts to isolate bacteria unique to the two soil types and then sequenced their 16S rRNA genes to determine their phylogenetic placement. Higher numbers of culturable bacteria, by over two orders of magnitude at the deepest sampling depths, were counted in the Anthrosols. Sequences of bacteria isolated on soil extract media yielded five possible new bacterial families. Also, a higher number of families in the bacteria were represented by isolates from the deeper soil depths in the Anthrosols. Higher bacterial populations and a greater diversity of isolates were found in all of the Anthrosols, to a depth of up to 1 m, compared to adjacent soils located within 50-500 m of their associated Anthrosols. Compared to standard culture media, soil extract media revealed diverse soil microbial populations adapted to the unique biochemistry and physiological ecology of these Anthrosols.

Fluorimetric determination of intra- and extracellular free amino acids in the microalgae Tetraselmis gracilis (Prasinophyceae) using monolithic column in reversed phase mode

This paper describes the development and application of an RP HPLC method using a C(18) monolithic stationary phase for the separation and quantification of extra- and intracellular amino acids in a batch cultivation of the marine alga Tetraselmis gracilis. Fluorimetric detection was made after separation of the o-phthaldialdehyde 2-mercaptoethanol (OPA-2MCE) derivatives using a binary gradient elution. Separation of 19 amino acids was achieved with resolution >1.5 in about 39 min at a flow rate of 1.5 mL/min. RSD of analyses in seawater medium ranged from 0.36% for Orn (0.50 mu mol/L) to 12% for Ile (0.10 mu mol/L). The main constituents of the intracellular dissolved free amino acids (DFAAs) in the exponential growth phase were arginine (Arg), asparagine (Asn), alanine (Ala), aspartic acid (Asp), glutamic acid (Glu), serine (Ser), glycine (Gly), glutamine (Gln), and leucine (Leu). The major amino acids excreted to the media were valine (Val), Ala, Ser, and Gly. The monolithic phase facilitates the analysis by shortening the separation time and saving solvents and instrumentation costs (indeed conventional HPLC instrumentation can be used, running at lower pressures than those ones used with packed particle columns).

Implementing stepwise solvent elution in sequential injection chromatography for fluorimetric determination of intracellular free amino acids in the microalgae Tetraselmis gracilis

The concept of sequential injection chromatography (SIC) was exploited to automate the fluorimetric determination of amino acids after pre-column derivatization with ophthaldialdehyde (OPA) in presence of 2-mercaptoethanol (2MCE) using a reverse phase monolithic C(18) stationary phase. The method is low-priced and based on five steps of isocratic elutions. The first step employs the mixture methanol: tetrahydrofuran: 10 mmol L(-1) phosphate buffer (pH 7.2) at the volumetric ratio of 8:1:91; the other steps use methanol: 10 mmol L-1 phosphate buffer (pH 7.2) at volumetric ratios of 20:80, 35:65, SO:SO and 65:35. At a flow rate of 10 mu L s(-1) a 25 mm long-column was able to separate aspartic acid (Asp), glutamic acid (Glu), asparagine (Asn), serine (Ser), glutamine (Gln), glycine (Gly), threonine (Thr), citruline (Ctr), arginine (Arg), alanine (Ala), tyrosine (Tyr), phenylalanine (Phe), ornithine (Orn) and lysine (Lys) with resolution >1.2 as well as methionine (Met) and valine (Val) with resolution of 0.6. Under these conditions isoleucine (Ile) and leucine (Leu) co-eluted. The entire cycle of amino acids derivatization, chromatographic separation and column conditioning at the end of separation lasted 25 min. At a flow rate of 40 mu L s(-1) such time was reduced to 10 min at the cost of resolution worsening for the pairs Ctr/Arg and Orn/Lys. The detection limits varied from 0.092 mu mol L(-1) for Tyr to 0.51 mu mol L(-1) for Orn. The method was successfully applied to the determination of intracellular free amino acids in the green alga Tetraselmis gracilis during a period of seven days of cultivation. Samples spiked with known amounts of amino acids resulted in recoveries between 94 and 112%. (C) 2008 Elsevier B.V. All rights reserved.

Agrobacterium tumefasciens-mediated transformation of the aquatic fungus Blastocladiella emersonii

Agrobacterium tumefaciens is widely used for plant DNA transformation and more recently, has also been used to transform yeast, filamentous fungi and even human cells. Using this technique, we developed the first transformation protocol for the saprobic aquatic fungus Blastocladiella emersonii, a Blastocladiomycete localized at the base of fungal phylogenetic tree, which has been shown as a promising and interesting model of study of cellular function and differentiation. We constructed binary T-DNA vectors containing hygromycin phosphotransferase (hph) or enhanced green fluorescent protein (egfp) genes, under the control of Aspergillus nidulans trpC promoter and terminator sequences. 24 h of co-cultivation in induction medium (IM) agar plates, followed by transfer to PYG-agar plates containing cefotaxim to kill Agrobacterium tumefsciens and hygromycin to select transformants, resulted in growth and sporulation of resistant transformants. Genomic DNA from the pool o resistant zoospores were shown to contain T-DNA insertion as evidenced by PCR amplification of hph gene. Using a similar protocol we could also evidence the expression of enhanced green fluorescent protein (EGFP) in zoospores derived from transformed cells. This protocol can also open new perspectives for other non-transformable closely related fungi, like the Chytridiomycete class. (C) 2011 Elsevier Inc. All rights reserved.

Structure and Applications of a Rhamnolipid Surfactant Produced in Soybean Oil Waste

Soybean oil soapstock was utilized as an alternative carbon source for the production of rhamnolipids by Pseudomonas aeruginosa LBI strain. The chemical composition and properties of the rhamnolipid mixture obtained were determined to define its potential applications. The chemical characterization of the rhamnolipid has revealed the presence of ten different homologues. The monorhamnolipid RhaC(10)C(10) and the dirhamnolipid Rha(2)C(10)C(10) were the main components of the mixture that showed predominance of 44% and 29%, respectively, after 144-h of cultivation. The biosurfactant was able to form stable emulsions with several hydrocarbons and showed excellent emulsification for soybean oil and chicken fat (100%). The rhamnolipid removed 67% of crude oil present in sand samples and presented antimicrobial activity against Bacillus cereus and Mucor miehei at 64 mu g/mL and inhibition of Neurospora crassa, Staphylococcus aureus, and Micrococcus luteus at 256 mu g/mL. The results demonstrated that the rhamnolipid produced in soybean oil soapstock can be useful in environmental and food industry applications.

Glädje i vårdandets värld

Föreliggande vårdvetenskapliga studie har som syfte att avtäcka och synliggöra glädjesom idé i vårdandets värld. Glädje sammanbinds med vårdandet genom kärleken somden tongivande kraften hos glädje. Vårdandets sak har genom historien alltid varit attlindra lidande och att tjäna liv och hälsa i en anda av kärlek och barmhärtighet. Dennastudie om glädje i vårdandet har en övergripande idéhistorisk inriktning och resultatetsammanställs i form av ett idémönster. Tolkningen sker genom en hermeneutisk ansatsmed fokus på förståelse av själva vårdandet. För att djupare förstå glädje, dessursprungsidé, väsen och uttrycksformer granskas begreppet ’glädje’ och de näraliggandebegreppen ’glad’ och ’ljus’ i etymologiska ordböcker samt i svenska, engelska ochlatinska ordböcker. Som stöd för tolkningen används klassiska texter innehållandefilosofers tankar om glädje. Glädje som idé glimtar fram i form av ett sjufaldigtmönster. Detta mönster innehåller särdrag hos glädje och det hjärtas natur somrespektive särdrag förverkligas i. I andlig mening utgör hjärtat livets medelpunkt ochkänslornas hemvist. Mönstret bildar bakgrund och blir vägledande vid den hermeneutiska läsningen omglädje, så som den framträder i berättelser om vårdande under åren 1900–1933. Dehistoriska källorna utgörs av facktidskriften Svensk sjukskötersketidning, böcker medberättelser om vårdande, arkivmaterial samt läroböcker om vårdarbete. Resultatetmynnar ut i ett idémönster, där idéer om det som gör glädje verksamt som vårdandeframträder. Dessa är det sanna hjärtats rena glädje – kärlek, glädje är ett kärleksbevis.Det brinnande hjärtats djupa glädje – livsglädje, glädje inspirerar och genererar kraft.Det bärande hjärtats glansfulla glädje – generositet, glädje är en gåva till den andra medlöfte om hjälp. Det inbjudande hjärtats glittrande glädje – gemenskap, glädje inbjudertill gemenskap. Det upprymda hjärtats uppsluppna glädje – integration, glädje gör attmänniskan kan glömma sitt lidande och närma sig den hon önskar vara. Detstämningsfulla hjärtats högtidliga glädje – bevärdigande, glädje skapar en anda ochatmosfär där människan upplever sig hedrad. Det fridfulla hjärtats stora glädje –räddning, en glädje visar sig när människan har erhållit det som kan begäras av gott ochundsluppet ont och är förnöjd med sin levnads lott. Förhoppningen är att dennagrundforskning ska öppna för ett nytt seende som kan leda till att glädjeuppmärksammas i vårdandets värld och artikuleras där.

To keep me safe from harm? Transgender prisoners and the experience of imprisonment

The experience of imprisonment for a transgender person is often a terrifying one. He or she is extremely vulnerable in such an environment from sexual violence from other prisoners. In addition, he or she may be exposed to inadequate or inappropriate medical care. Consequently transgender prisoners are often denied the protection offered by role of law. A significant reason for this treatment is the erasure of the transgender experience in informing the nature of the prison regime. In particular, the failure to give sufficient weight to gender self identification by transgender prisoners exposes them to risks which other prisoners do not have to endure. It is suggested that the only way to reduce such harm is through the cultivation of a prison regime based upon the lives of transgender prisoners.

Bringing shared values out of the shadows

As part of its wider promotion of a world that is peaceful and tolerant, the United Nations declared 2005 the International Year of Sport and Physical Education. At the fore of the UN's proclamation was recognition of the important role sport and PE play in nurturing the health and harmony of society. Sport and PE, the UN declared, provide important nodes for social connection around which shared values and understandings can be formed. In the wake of the UN's endorsement of the role that sport and PE can play in fostering social and emotional development, it is more important that ever that we reflect on and refine our practices towards this end. This paper draws on two research narratives to illustrate how easily this potential can be undermined. Indeed, behind some of the wonderful opportunities for connection that exist through a participation in PE and sport exists a dark shadow of alienation and oppression. The challenge raised through this paper centres on the need for PE teachers and coaches to work deliberately and strategically towards the cultivation of inclusive, tolerant and responsible forms of participation, and not leave their development to chance.