RESUMO
OBJECTIVE: To describe and analyze a new protocol for the extraction of platelet-rich plasma (PRP) for use in clinical practice and compare this technique with methods that have been previously described in the medical literature. METHODS: We extracted PRP from 20 volunteers using four different protocols (single spin at 1600 ×g, single spin at 600 ×g, double spin at 300 and 700 ×g, and double spin at 600 and 900 ×g). In another group of 12 individuals, we extracted PRP with our new technique (named 'turn down-turn up') consisting of a double spin (200 ×g and 1600 ×g) closed system using standard laboratory equipment (including an ordinary benchtop centrifuge), where the blood remained in the same tube during all processes, reducing the risk of contamination. Platelet counts adjusted to baseline values were compared using analysis of covariance (ANCOVA). RESULTS: Using the four previously described protocols (mentioned above), we obtained concentrations of platelets that were 1.15-, 2.07-, 2.18-, and 3.19-fold greater than the baseline concentration, respectively. With the turn down-turn up technique, we obtained a platelet count that was 4.17-fold (95% confidence interval (CI): 3.09 to 5.25) greater than the baseline platelet count (p=0.063 compared with the double spin at 600 and 900 ×g method). The total cost of the disposable materials used in the extraction process was less than US$10.00 per individual. CONCLUSION: In the present study, we described a simple and safe method for obtaining PRP using low-cost devices.
Assuntos
Centrifugação/métodos , Técnicas de Laboratório Clínico/métodos , Plasma Rico em Plaquetas , Adulto , Centrifugação/economia , Centrifugação/normas , Técnicas de Laboratório Clínico/economia , Técnicas de Laboratório Clínico/normas , Análise Custo-Benefício , Humanos , Masculino , Pessoa de Meia-Idade , Contagem de Plaquetas , Reprodutibilidade dos Testes , Fatores de TempoRESUMO
OBJECTIVE: To describe and analyze a new protocol for the extraction of platelet-rich plasma (PRP) for use in clinical practice and compare this technique with methods that have been previously described in the medical literature. METHODS: We extracted PRP from 20 volunteers using four different protocols (single spin at 1600 ×g, single spin at 600 ×g, double spin at 300 and 700 ×g, and double spin at 600 and 900 ×g). In another group of 12 individuals, we extracted PRP with our new technique (named 'turn down-turn up') consisting of a double spin (200 ×g and 1600 ×g) closed system using standard laboratory equipment (including an ordinary benchtop centrifuge), where the blood remained in the same tube during all processes, reducing the risk of contamination. Platelet counts adjusted to baseline values were compared using analysis of covariance (ANCOVA). RESULTS: Using the four previously described protocols (mentioned above), we obtained concentrations of platelets that were 1.15-, 2.07-, 2.18-, and 3.19-fold greater than the baseline concentration, respectively. With the turn down-turn up technique, we obtained a platelet count that was 4.17-fold (95% confidence interval (CI): 3.09 to 5.25) greater than the baseline platelet count (p=0.063 compared with the double spin at 600 and 900 ×g method). The total cost of the disposable materials used in the extraction process was less than US$10.00 per individual. CONCLUSION: In the present study, we described a simple and safe method for obtaining PRP using low-cost devices.
Assuntos
Humanos , Masculino , Adulto , Pessoa de Meia-Idade , Centrifugação/métodos , Técnicas de Laboratório Clínico/métodos , Plasma Rico em Plaquetas , Contagem de Plaquetas , Fatores de Tempo , Centrifugação/economia , Centrifugação/normas , Reprodutibilidade dos Testes , Análise Custo-Benefício , Técnicas de Laboratório Clínico/economia , Técnicas de Laboratório Clínico/normasRESUMO
Studies on free-living amoebae (FLA), has been increased in recent years, especially related to the genus Acanthamoeba, because these organisms are widely found in the environment. The present work isolated and characterized this organism from biofilms and dust in hospital environment. 135 samples were collected in 15 different environments in a hospital at the south of Brazil. Thirty-one (23%) isolates were identified as morphologically belonging to the Acanthamoeba genus and 10 of these were submitted to temperature and osmotolerance tests as criterion for evaluation of the viability and pathogenicity. The tests indicate that four (40%) of these isolates could be potentially pathogenic because grew at high temperature (40 degrees C) and osmolarity (mannitol 1 M). Some isolates genotypes were determined after ribosomal DNA sequencing. These data revealed that three dust isolates belong to T4, two biofilm isolates to T5 and one dust isolate to T3 genotype. Therefore, Acanthamoeba found in the hospital environment represents a risk for people that circulate there.
Assuntos
Acanthamoeba/isolamento & purificação , Acanthamoeba/patogenicidade , Acanthamoeba/genética , Acanthamoeba/fisiologia , Animais , Biofilmes , Brasil , Criança , DNA Ribossômico/genética , Poeira/análise , Serviço Hospitalar de Emergência/normas , Flagelos/fisiologia , Genótipo , Hospitais Públicos , Humanos , Unidades de Terapia Intensiva/normas , Manitol/metabolismo , Reação em Cadeia da Polimerase , RNA Ribossômico 18S/genética , Centros Cirúrgicos/normasRESUMO
A study on the presence of free-living amoebae in a public hospital was developed in the city of Porto Alegre, State of Rio Grande do Sul. Dust and biofilms were collected using sterile swabs that had been prepared for this study, from 15 hospital environments, including the intensive care center, pediatric intensive care unit, kitchen, emergency room, outpatient surgical center, clinical surgical center, water storage tanks, taps and six drinking fountains for general use, every month from July 2004 to March 2005. The FLAs were isolated by culturing, using non-nutrient agar medium with the addition of heat-killed Escherichia coli. The protozoa were identified by morphological observation of cysts and trophozoites, in accordance with Page's morphological criteria (1988). Among the 135 samples collected from the 15 environments, 47 (35%) were positive for FLAs. Of these, thirty-four percent presented morphological characteristics particular to the genus Acanthamoeba.
Assuntos
Acanthamoeba/isolamento & purificação , Amoeba/isolamento & purificação , Biofilmes , Poeira , Acanthamoeba/fisiologia , Amoeba/classificação , Amoeba/fisiologia , Animais , Brasil , Hospitais Públicos , HumanosRESUMO
Um estudo sobre a presença de amebas de vida livre em um hospital público foi desenvolvido na Cidade de Porto Alegre, RS. Poeira e biofilmes de 15 ambientes hospitalares, incluindo CTI, UTI pediátrica, cozinha, emergência, centro cirúrgico ambulatorial e centro cirúrgico, reservatórios de água, torneira e 6 bebedouros coletivos foram coletados mensalmente, de julho de 2004 a março de 2005, usando-se suabes estéreis, preparados para a pesquisa. As AVL foram isoladas em cultivo, utilizando-se meio de ágar não nutriente adicionado de Escherichia coli, mortas pelo calor. A identificação dos protozoários foi feita pela observação morfológica de cistos e trofozoítos, segundo critérios morfológicos de Page (1988). Das 135 amostras coletadas dos 15 ambientes estudados, 47 (35 por cento) foram positivas para AVL. Destas, 34 por cento apresentaram características morfológicas próprias do gênero Acanthamoeba.
A study on the presence of free-living amoebae in a public hospital was developed in the city of Porto Alegre, State of Rio Grande do Sul. Dust and biofilms were collected using sterile swabs that had been prepared for this study, from 15 hospital environments, including the intensive care center, pediatric intensive care unit, kitchen, emergency room, outpatient surgical center, clinical surgical center, water storage tanks, taps and six drinking fountains for general use, every month from July 2004 to March 2005. The FLAs were isolated by culturing, using non-nutrient agar medium with the addition of heat-killed Escherichia coli. The protozoa were identified by morphological observation of cysts and trophozoites, in accordance with Page's morphological criteria (1988). Among the 135 samples collected from the 15 environments, 47 (35 percent) were positive for FLAs. Of these, thirty-four percent presented morphological characteristics particular to the genus Acanthamoeba.
Assuntos
Humanos , Animais , Acanthamoeba/isolamento & purificação , Amoeba/isolamento & purificação , Biofilmes , Poeira , Amoeba/classificação , Brasil , Hospitais PúblicosRESUMO
Análises coproparasitológicas foram realizadas em 222 crianças de um loteamento da periferia de Porto Alegre-RS, entre agosto de 2004 e setembro de 2005, pelo método de Hoffman, Pons e Janner. A positividade para enteroparasitos foi de 102 (46por cento) amostras. A faixa etária de 12 a 14 anos foi a que apresentou maior percentual de positividade (58,5por cento). O parasito mais freqüente foi Trichuris trichiura (18,9por cento), seguido por Ascaris lumbricoides e pelo comensal Entamoeba coli (ambos com ocorrência de 16,7por cento), Giardia lamblia (2,7por cento), Strongyloides stercoralis (4,5por cento), Hymenolepis nana (2,7por cento) e Enterobius vermicularis (2,2por cento). O monoparasitismo foi observado em 63,7por cento dos casos positivos. A associação entre A. lumbricoides e T. trichiura ocorreu 16 vezes no estudo. Os resultados obtidos apontam para a necessidade contínua de investimentos dos setores públicos em saúde e infra-estrutura somados a investimentos em educação e treinamento de educadores visando a uma melhor aplicação dos conhecimentos sobre a prevenção das parasitoses e à conseqüente melhora na qualidade de vida da população.