Cultivo celular in vitro: importância para a pesquisa biomédica e dimensão da problemática de autenticação de linhagens celulares

Experimental models composed by human and animal cell lines are simplified and informative, allowing them to be widely used for biomedical research. Most laboratories that use in vitro cultivated cells maintain a variation of cell lines stored and cultivated. Therefore, misidentification and cross-contamination events can happen during cell lines handling. This problem can generate a repertoire of dubious results and papers, which may prejudice biomedical research. Recently it was created the International Cell Line Authentication Committee (ICLAC), which aims to spread knowledge about cross-contamination and misidentification of in vitro cell lines. Despite of the efforts spent trying to aware scientific community about the importance of the correct identification of cells, the number of papers based on misidentified cell lines it´s still worrying, compromising the reliability of out coming results and conclusions regarding them. The present study aims to analyze and discuss the main advantages and limitations of eukaryote in vitro cell lines use, characterizing the cell lines authentication problems. Therefore, compilation and critical analyses of literature data was realized, aiming to improve the understanding about this subject. Based on information about 445 cell lines with issues published by ICLAC it´s clear that contamination in human cell lines represented 89,2 % of mentioned problems. HeLa cell line was the responsible for most contamination, especially in 92 normal tissue cell lines, representing 44,6% of the contamination. These results reinforce the importance of periodic maintenance of cell lines cultures by labs and implementation of authentication methods as polymorphic STRs, besides obtaining cell lines from reliable sources and cell banks

Avaliação do potencial anti-inflamtório de extratos isolados de microrganismos marinhos

O Brasil possui cerca de 8.000 quilômetros de costa litorânea, a segunda maior do mundo depois da Austrália. Apesar do território marítimo brasileiro apresentar 3,6 milhões de quilômetros quadrados de extensão, as pesquisas relacionadas ao levantamento de potenciais recursos brasileiros obtidos a partir de espécies marinhas ainda são incipientes. O interesse de empresas farmacêuticas por compostos isolados de microrganismos vem crescendo lentamente devido às dificuldades até então encontradas para o isolamento e cultivo dos mesmos. No entanto, com o avanço destas técnicas, recentes estudos demonstram que compostos isolados de microrganismos marinhos representam uma rica e ainda pouco explorada fonte de novos compostos bioativos (estima-se que apenas 1% dos microrganismos marinhos foram cultivados e identificados). Recentes estudos que apontam para o potencial farmacológico de compostos isolados de microrganismos nos instigaram à busca de novos fármacos com potencial imunoestimulador não apenas a partir de invertebrados, mas também de microrganismos marinhos e seus metabólitos secundários bioativos. Neste trabalho, após avaliação do potencial imunomodulador de extratos brutos derivados de microrganismos marinhos, obteve-se a substância pura Pirenocine A, isolada do fungo Penicillium paxilli, que mostrou-se promissora devido sua capacidade de diminuir a produção de óxido nítrico e citocinas pró-inflamatórias em macrófagos estimulados com LPS tanto no protocolo profilático quanto terapêutico ...

Cinética de crescimento do fungo termófilo Myceliophthora thermophila M. 7.7 em cultivo no estado sólido

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Desenvolvimento do cultivo 3D a partir de células primarias de neoplasias mamárias caninas: estudo da apoptose sobre efeito ou não da carboplatina

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Família Enterobacteriaceae, Acinetobacter baumannii e Pseudomonados na microbiota bucal de pacientes mantidos em unidades de terapia intensiva

Enteric organisms, pseudomonads and other opportunistic microorganisms in the oral microbiota have been linked to serious infections in patients hospitalized in intensive care units (ICU). The present study evaluated the presence of family Enterobacteriaceae, Pseudomonas aeruginosa, and Acinetobacter baumannii in the mouth of patients in ICU, correlating it with oral and systemic conditions. Data on health, socioeconomic status, medication use, drug addiction, medical and family histories of patients held for more than 72 hours in the ICU with a diagnosis of severe infection or that developed this condition after entry in said unit were obtained. Fifty patients provided clinical samples of supragingival and subgingival biofilms, saliva and oral mucous membranes were collected, as well as respiratory secretions from patients with pneumonia, blood and urine for sepsis. The presence of target microorganisms was carried out by polymerase chain reaction (PCR) and by culture using selective media. The Chi-square and Mann-Whitney tests were used for statistical analysis, and the significance level was 5%. The intraoral clinical conditions of the patients were poor. The family Enterobacteriaceae was the most prevalent, affecting 39.5% of the supragingival biofilm samples of patients attended in ICU and 18.6% of patients in the control group, besides the rods were the only group found in extraoral samples.

Enterococcus spp. em espécimes clínicos de pacientes atendidos em unidades de terapia intensiva

The relationship between the occurrence of enterococci in the oral microbiota and serious infections in patients hospitalized in intensive care units (ICU) has been established. This study evaluated the presence of Enterococcus faecalis and other species of this genus in the mouths of patients on ICU, correlating it with oral and systemic conditions. Data on health and socioeconomic, medication use, medical and family history of patients maintained for 72 hours in the ICU, diagnosed with severe infection or who have developed this condition after the entry to the unit were obtained. Fifty patients provided intraoral and extraoral clinical samples for analysis (above and subgingival biofilm, saliva and buccal mucosa, followed by obtaining samples of respiratory secretions for patients with pneumonia, and blood and urine for sepsis). The presence of target microorganisms was performed by polymerase chain reaction (PCR) and culture using selective media. The chi-square and Mann-Whitney tests were used for statistical analysis, and the significance level was 5%. The intraoral clinical conditions of the patients showed poor. E. faecalis was significantly more frequent microorganism, followed by E. faecium. The use of broadspectrum antimicrobial action was associated with the presence of these opportunistic microorganisms. These bacteria were more frequent in patients with periodontitis or gingivitis. The results showed that enterococci associated with serious infectious processes may originate from resident microbiota of patients and its prevalence is not elevated in healthy individuals.

Opportunistic microorganisms in patients with head and neck trauma

The use of antimicrobial agents has facilitated the dissemination of multi-resistant microorganisms, compromising dental and medical treatment. The aim of this study was to evaluate the distribution of different opportunistic microbial species in patients who suffered head and neck trauma, under temporary maintenance in nosocomial environment, particularly intensive care units, on the occurrence of such microorganisms in the oral cavity of the patients. It was selected 38 patients subjected to head and neck traumas. After emergency surgical procedures, clinical samples of saliva, sub and supragingival biofilms and mucosal surfaces were collected at two different moments: just after stabilization of the patient and soon after patients’ release from medical units. The presence of opportunistic and superinfecting microorganisms was evaluated by culture on selective and non-selective media, and the presence of the family Enterobacteriaceae, as well as genera Enterococcus, Pseudomonas, and Staphylococcus was assessed by PCR. It was found that the use of antimicrobials, even for short periods of time was sufficient to facilitate colonization by microorganisms of the families Enterobacteriaceae and Pseudomonadaceae, as well as yeasts and enterococci. These results support the concept that medical and dental teams should make a periodically change of antimicrobials used in treatment protocols in hospital for head and neck trauma patients, in order to minimize dissemination of opportunistic or superinfecting microorganisms.

Fatores microbiológicos e imunológicos envolvidos no desenvolvimento da cárie precoce da infância

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Uso de resíduos madeireiros da Amazônia brasileira no cultivo in vitro de Lentinus strigosus

Lentinus strigosus (Schwein.) Fr. is an exploitable edible mushroom occurring in the Brazilian Amazon, being part of a huge diversity of edible mushrooms which are little grown. The use of regional waste is recommended to reduce production costs of any kind of edible mushroom. Thus, the mycelial growth of L. strigosus in culture media based on regional wood waste extract by using substrates based on Protium puncticulatum, Cariniana micrantha and Caryocar glabum sawdust, supplemented with 20% of wheat bran (Triticum aestivum), corn bran (Zea sp.) or rice bran (Oryza sp.) was observed. Eucalyptus (Eucaliptus sp.) sawdust was used for comparison with the other wood wastes because it is commonly used in the cultivation of edible fungi. The experimental design employed was totally randomized, in 4 x 3 factorial scheme (sawdust x bran), adding up 12 treatments with 5 repetitions, being that each repetition corresponded to a Petri dish, totalizing 60 dishes, incubated at 35 ºC. The diameter of the colony was daily evaluated until the fungus reached the borders of the Petri dish in one of the treatments. After that period, the media based on P. puncticulatum sawdust obtained thebest results of mycelial growth, showing potential to be used as an alternative residuein a future production of L. strigosus in the state of Amazonas.

Resíduos de bananeira como substrato base para o cultivo in vitro de Coprinus comatus

Coprinus comatus is an edible and lignolitic fungus which has presented great potential for commercial use due to its easy development in the different residues, such as banana tree leave. Thus, the mycelial growth of Coprinus comatus in culture media based on leaves of Thap-Maeo, Prata-Anã, Pelipita and Caipira banana tree cultivars, supplemented with 20% of wheat, soy and rice brans, was evaluated. 7 mm-wide discs of CCO 01/01 strain of C. comatus were inoculated in the middle of Petri dishes containing culture medium, inside a laminar flow chamber. Next, the dishes were arranged totally at random inside an incubator at 25 ºC. The daily measurements of the mycelial growth began after 24 hours, until one of the treatments reached the borders of the Petri dish. According to the results obtained, we verified that there was not effect of the kind of supplementation for culture media based on Thap-Maeo, Prata-Anã and Pelipita; the best growth averages for culture media based on Caipira were provided by wheat and rice brans. Therefore, banana residues may be a viable and ecologically correct choice for the cultivation of C. comatus, especially for Thap-Maeo and Prata Anã sorts, which provided the best growth averages, regardless of the supplementation used.

Cultivo in vitro de Pleurotus ostreatus em resíduos de bananeira

The objective of this paper was to evaluate the mycelial growth of Pleurotus ostreatus (strain POS 09/100) in culture media based on different banana tree residues. The experimental design was totally randomized in 3 x 4 factorial scheme and consisted in three combinations of residues (pseudostem, leave and pseudostem + leave) and four banana tree cultivars (Thap Maeo, Prata Anã, Pelipita and Caipira), totalizing twelve treatments each with five repetitions, adding up sixty experimental units. Growth was measured every 24 hours until the mycelium of one of the treatments reached the border of the Petri dish, what occurred five days after the beginning of the experiment. The results obtained showed that all the combinations of banana tree residues were favorable to P. ostreatus mycelial growth, especially pseudostem + leaf of Pelipita, Thap maeo and Prata anã cultivars. Thus, the use of banana tree residues is viable for cultivation of P. ostreatus, and considered as an excellent alternative, besides reducing their disposal in the environment.

Cultivo de Lentinula edodes e Pleurotus ostreatus em bagaço de cana-de-açúcar

Pós-graduação em Microbiologia - IBILCE

Produção de conídios do fungo entomopatogênico Metarhizium anisopliae em diferentes condições de cultivo e em biorreator de bandeja

Pós-graduação em Microbiologia - IBILCE

Produção de conídios do fungo entomopatogênico Metarhizium anisopliae em diferentes condições de cultivo e em biorreator de bandeja

Pós-graduação em Microbiologia - IBILCE

Desarrollo y validación de tecnologías de intensificación lumínica para fotobiorreactores (FBR) para el aprovechamiento de la radiación en el cultivo de microalgas

En la actualidad la humanidad se enfrenta a una serie de problemas de gran transcendencia que van a determinar en alto grado los niveles de calidad de vida en los próximos años. El calentamiento global, el incremento demográfico incontrolado, la contaminación ambiental y la escasez de recursos así como una mala distribución de los mismos, son sólo algunos ejemplos. En este contexto, las microalgas, microrganismos fotosintéticos de alta eficiencia y versatilidad, presentan una serie de características que las convierten a priori en la base de una tecnología con un enorme potencial para formar parte de la solución a estos graves problemas planteados. Uno de los principales factores que impiden una mayor implantación de la tecnología de microalgas es de tipo económico. La baja productividad por unidad de área de los sistemas de cultivo actuales y la alta inversión necesaria en equipos y mantenimiento, hace que solo se justifique el cultivo de productos de muy alto valor añadido. Las soluciones pasan por aumentar el rendimiento global de los cultivos y por disminuir los costes de equipos e instalaciones. La presente tesis investiga sobre la posibilidad de conseguir un mejor aprovechamiento de la luz solar incidente sobre un cultivo de microalgas mediante el uso de una serie de dispositivos ópticos que vienen a denominarse intensificadores lumínicos. De entre los factores que determinan la productividad de un campo de cultivo de microalgas, posiblemente el más determinante sea la cantidad de radiación que las microalgas pueden aprovechar. Los intensificadores aumentan la disponibilidad de luz en el interior de los tubos de cultivo, de forma que la fotosíntesis se vea favorecida y, de este modo, se incremente la tasa de crecimiento de las microalgas. En el desarrollo de la tesis se proponen tres tipologías diferentes de intensificador diseñadas en base a criterios óptico-geométricos. Para cada una de estas tipologías se evalúa el incremento de radiación que se lograría sobre un tubo de cultivo. Paralelamente se desarrolla un modelo que permite la evaluación de la productividad del cultivo. Esto permite añadir el factor biológico al puramente óptico-físico y valorar las distintas propuestas de intensificadores en función de las características propias del microalga utilizada en el cultivo. El modelo es verificado y contrastado frente a datos de producción obtenidos en la bibliografía. Finalmente, la exposición concluye con una presentación general de las futuras líneas de investigación. ABSTRACT Today, humanity is facing a series of problems of global significance that will determine the standard of living in the years ahead. Global warming, uncontrolled population growth, pollution, lack of resources and poor distribution of them are just an example of the challenges we are facing. In this context, microalgae, high efficient photosynthetic microorganisms, have a number of characteristics that turn them into a very promising technology that can contribute or be part of a sustainable solution. One of the main factors preventing the adoption of microalgae technology is economical. The low productivity per unit area of current farming systems and the high investment needed in equipment and maintenance, only justifies the cultivation of algae for high value applications. One solution could be increasing the overall yield of the crops and reduce the equipment and facilities costs. Among the factors that determine the productivity of a microalgae culture, possibly the most influential one is the amount of radiation that microalgae receive. This Thesis develops the possibility of making better use of the sunlight radiation incident on a crop field using a series of devices similar to solar collectors. The solar collectors proposed are intended to increase the availability of light inside the culture tubes and within it, the tax of photosynthesis and the overall growth rate of the microalgae. In this research, three different configurations of collectors are designed, based on optical and geometrical criteria. For each one of these collectors, the increase on radiation that would be expected is evaluated. Furthermore, a model for light distribution inside the culture is developed in terms to estimate the biomass productivity. This allows adding the biological factor to purely optical-physical considerations and to assess the different solar collectors proposed, in terms specific for the microalgae. The model is tested against production data obtained in different scientific literature. Finally, the exposition concludes with some guidelines for future research.