Made available in DSpace on 2014-06-11T19:23:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-05-04Bitstream added on 2014-06-13T20:30:10Z : No. of bitstreams: 1 camargo_la_me_arafcf.pdf: 458749 bytes, checksum: e566ad21072ed4474151776ec23f5806 (MD5) / Universidade Estadual Paulista (UNESP) / A presente dissertação permitiu quantificar a presença de glicerol-3-fosfato oxidase (GPO, sn-glicerol-3-fosfato: oxigênio 2-oxidorredutase, EC 1.1.3.21) em extratos de levedura seca de panificação por dois métodos: polarográfico e colorimétrico. A melhor metodologia de purificação da GPO foi obtida por rompimento celular com esferas de vidro, em homogenizador do tipo Bead Beater (Biospec products, USA), por 15 minutos, com 27,6% de eficiência de lise celular. O extrato celular bruto foi tratado com 1% de sulfato de estreptomicina antes da precipitação com igual volume de solução 30% (p/v) de polietilenoglicol 3350, dialisado e a sua atividade otimizada por método colorimétrico. A determinação das características da GPO foi possível em ensaios contendo: 250 mM de glicerol-3-fosfato em tampão 0,1 M Tris-HCl pH 8,0 contendo 0,1% Triton X-100; 0,0133% de 4-aminoantipirina; 0,0266% de fenol; cerca de 0,40 unidade de peroxidase (PO) e água destilada para completar o volume de ensaio. A reação foi iniciada pela adição de 15 æL de extrato enzimático diluído 10 vezes seguido de uma incubação de 2 horas a 60°C e interrompida pela adição de solução 10% de SDS e a coloração desenvolvida foi medida a 500 nm. A GPO apresentou alta estabilidade térmica, pH de estabilidade entre 7,0 - 8,0 e a presença de azida de sódio na concentração de 0,05% manteve a atividade da enzima por 21 dias a 40°C. Este método permitiu também quantificar glicerol-3- fosfato, importante metabólito intermediário da biossíntese lipídica e glicolítica, na faixa de 56 - 250 mM. / The present dissertation allowed to quantify the presence of glycerol-3- phosphate oxidase (GPO, sn-glycerol-3-phosphate: oxygen 2-oxidoreductase, EC 1.1.3.21) in baker s yeast extract by two methods: polarographic and colorimetric. The best methodology of purification of GPO was obtained by cell debris with glass beads, in a Bead Beater homogenizator (Biospec products, USA), for 15 minutes, with 27.6% of efficiency of cellular lysis. The crude cellular extract was treated with 1% of streptomycin sulfate before the precipitation, with equal volume of a solution of 30% (w/v) polyethylene glycol 3350, dialysed and its activity was optimized by colorimetric method. The determination of the characteristics of GPO was possible in assays containing: 250 mM of glycerol-3-phosphate in 0.1 M Tris-HCl buffer, pH 8.0 containing 0.1% Triton X-100; 0.0133% of 4-aminoantipyrine; 0.0266% of phenol; about 0.40 unit of peroxidase (PO) and water distilled to complete the volume. The reaction was started by the addition of 15 ìL of enzymatic extract diluted 10 times, followed by incubation of 2 hours at 60°C, interrupted by the addition of solution 10% of SDS, and the developed coloration was measured at 500 nm. GPO presented high thermal stability, pH of stability among 7.0 - 8.0, and the presence of sodium azide in the concentration of 0.05% maintained the activity of the enzyme for 21 days at 40°C. This method also allowed to quantify glicerol-3- phosphate, important intermediate metabolite of lipid biosynthesis and glycolysis, in the range 56 - 250 mM.

Resumo: A presente dissertação permitiu quantificar a presença de glicerol-3-fosfato oxidase (GPO, sn-glicerol-3-fosfato: oxigênio 2-oxidorredutase, EC 1.1.3.21) em extratos de levedura seca de panificação por dois métodos: polarográfico e colorimétrico. A melhor metodologia de purificação da GPO foi obtida por rompimento celular com esferas de vidro, em homogenizador do tipo Bead Beater (Biospec products, USA), por 15 minutos, com 27,6% de eficiência de lise celular. O extrato celular bruto foi tratado com 1% de sulfato de estreptomicina antes da precipitação com igual volume de solução 30% (p/v) de polietilenoglicol 3350, dialisado e a sua atividade otimizada por método colorimétrico. A determinação das características da GPO foi possível em ensaios contendo: 250 mM de glicerol-3-fosfato em tampão 0,1 M Tris-HCl pH 8,0 contendo 0,1% Triton X-100; 0,0133% de 4-aminoantipirina; 0,0266% de fenol; cerca de 0,40 unidade de peroxidase (PO) e água destilada para completar o volume de ensaio. A reação foi iniciada pela adição de 15 æL de extrato enzimático diluído 10 vezes seguido de uma incubação de 2 horas a 60°C e interrompida pela adição de solução 10% de SDS e a coloração desenvolvida foi medida a 500 nm. A GPO apresentou alta estabilidade térmica, pH de estabilidade entre 7,0 - 8,0 e a presença de azida de sódio na concentração de 0,05% manteve a atividade da enzima por 21 dias a 40°C. Este método permitiu também quantificar glicerol-3- fosfato, importante metabólito intermediário da biossíntese lipídica e glicolítica, na faixa de 56 - 250 mM. / Abstract: The present dissertation allowed to quantify the presence of glycerol-3- phosphate oxidase (GPO, sn-glycerol-3-phosphate: oxygen 2-oxidoreductase, EC 1.1.3.21) in baker’s yeast extract by two methods: polarographic and colorimetric. The best methodology of purification of GPO was obtained by cell debris with glass beads, in a Bead Beater homogenizator (Biospec products, USA), for 15 minutes, with 27.6% of efficiency of cellular lysis. The crude cellular extract was treated with 1% of streptomycin sulfate before the precipitation, with equal volume of a solution of 30% (w/v) polyethylene glycol 3350, dialysed and its activity was optimized by colorimetric method. The determination of the characteristics of GPO was possible in assays containing: 250 mM of glycerol-3-phosphate in 0.1 M Tris-HCl buffer, pH 8.0 containing 0.1% Triton X-100; 0.0133% of 4-aminoantipyrine; 0.0266% of phenol; about 0.40 unit of peroxidase (PO) and water distilled to complete the volume. The reaction was started by the addition of 15 ìL of enzymatic extract diluted 10 times, followed by incubation of 2 hours at 60°C, interrupted by the addition of solution 10% of SDS, and the developed coloration was measured at 500 nm. GPO presented high thermal stability, pH of stability among 7.0 - 8.0, and the presence of sodium azide in the concentration of 0.05% maintained the activity of the enzyme for 21 days at 40°C. This method also allowed to quantify glicerol-3- phosphate, important intermediate metabolite of lipid biosynthesis and glycolysis, in the range 56 - 250 mM. / Orientador: Edwil Aparecida de Lucca Gattás / Coorientador: Maristela de Freitas Sanches Peres / Banca: Valdir Augusto Neves / Banca: Maria de Lourdes T. de Moraes Polizeli / Mestre

Group Purchasing Organizations (GPOs) play a significant role in the healthcare industry. The presence of GPOs helps the healthcare centers to offload their responsibilities so that they can focus on more critical areas which require attention like providing quality care. This thesis involves the comparison of three models of procurement operations in terms of cost efficiency. This cost comparison model features a healthcare organization associated with a national GPO, a healthcare organization which procures by self sourcing (not associated with a GPO), and a hybrid procurement model involving a national GPO and a regional GPO. The comparison model highlighted the cost effectiveness of these three different ways of procurement, which threw significant light on the purchasing operations of healthcare organizations. In the second part of this research study, we formulated a method to measure the degree of access to innovative products across the above mentioned procurement models either involving on-contract (from a GPO) purchasing, or off-contract purchasing (from individual manufacturers not affiliated to GPO) or both. We also identified the metrics for innovation and measure the innovativeness of products. Based on the literature study, it was found that purchasing groups may also be an entry barrier to new suppliers (Zweig 1998), with big national GPOs dominating the market and dictating the pricing of commodities. The first hypothesis H1 of this research study was stated as "National GPOs (Group Purchasing Organizations) enable the healthcare establishments to lower the cost of medical services and operations." The second hypothesis H2 of this research study was acknowledged as "National GPOs a barrier to entry of Innovative product manufacturers in the healthcare industry." This thesis will identify the advantages and disadvantages of each type of procurement operation and address the economic issues which affect the relationship between a healthcare center and a GPO. The proposed research would indirectly help to identify whether cost savings are being shared by the links in the downstream supply chain and the savings are being percolated to the patients for the added welfare of the society. It will also identify the importance of innovative products in the society and will raise the bar of specialty treatments without compromising on the level of service being offered to the patients. This thesis will also highlight positive aspects of niche manufacturers of innovative products with smaller volumes which are currently marginalized in the market by the big national players. To the best of the author's knowledge, the research objective of measuring innovation of products has not been addressed yet in academic literature and will have the benefit of comparing three different purchasing models used in healthcare industry.

Apples datorer med operativsystemet OS X blir allt vanligare på arbetsplatserna. För att företagets IT-avdelning ska kunna upprätthålla en stabil och säker miljö strävar man efter att standardisera system och hårdvara. Verkligheten stämmer inte alltid överens med IT-avdelningens strävan och anställda vill i ökande grad använda sig av produkter de valt själva. Arbetet har undersökt möjligheterna och begränsningarna med att integrera Apple OS X i Microsofts Active Directory. För att kunna besvara de frågor som använts som avgränsning inleddes arbetet med att metodiskt studera litteratur och på internet publicerade artiklar. Därefter har laborationer med och utan tredjepartsmjukvarorna ADmitMac och Centrify genomförts med inriktning på arbetets avgränsningar. Parallellt med detta har intervjuer av organisationer med olika bakgrund genomförts. Intervjuer och laborationer gav ett resultat som sedan diskuterades i rapportens avslutande del. I analysen gjordes en jämförelse av resultaten från intervjuer samt laborationer. Från detta kunde bland annat följande slutsats dras; För att integrera en dator med Apple OS X i Active Directory fungerar det inbyggda stödet men med begränsad funktionalitet. Med hjälp av tredjepartsmjukvaror utökas stödet och fler funktioner i Active Directory kan användas.

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