Análise do efeito in vitro de acridonas na replicação do Vírus da Hepatite C

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influência de Lactobacillus rhamnosus na patogenicidade e na expressão de genes de virulência de Candida albicans: estudo in vitro e in vivo

Pós-graduação em Biopatologia Bucal - ICT

Efeitos de reguladores do metabolismo lipídico no desenvolvimento in vitro de embriões bovinos e na sobrevivência à vitrificação

Pós-graduação em Medicina Veterinária - FCAV

Principais mecanismos envolvidos na maturação oocitária em bovinos: da oogenese a maturação in vitro

Despite the efforts made to improve the production of bovine embryos in vitro, their efficiency is still low, since only 30-40% of developed blastocysts are obtained from oocytes after in vitro maturation (IVM), fertilization and cultured embryos. Assisted reproductive technologies have a limiting impact due a lack of oocytes capable to fertilization.The comprehension of mechanism involved in oocyte maturation are crucial to establish a culture system that allows a larger number production of good quality embryos. The study of the early stages of oocyte and follicle development in vivo is important for a better understanding of the molecular pathways that regulate oogenesis, folliculogenesis and oocyte maturation. Thus the physiological biochemical and molecular mechanisms involved in maturation may contribute to the increased efficiency of in vitro embryo production. Therefore, the aim of this literature review is to understand the basic mechanisms that underlie oocyte maturation in cattle, since oocyte and follicle cells in vivo formation to its use in the in vitro environment.

In vitro production of bovine embryos using Sigma antioxidant supplement®, α-tocopherol and L-ascorbic acid

This study tested the effect of Sigma antioxidant supplement®, α-tocopherol (vitamin E) and L-ascorbic acid (vitamin C) in the culture medium of bovine embryos. In experiment 1, in vitro produced bovine zygotes were cultured in Human Tubal Fluid (HTF): Eagle’s Basic Medium (BME) with: Group 1 – 50 µm vitamin C; Group 2 – 200 µm vitamin E; Group 3 – 25 µm vitamin C and 100 µm vitamin E; Group 4 – 1 µl/ml Sigma antioxidant supplement®; and the Control group – HTF:BME only. In experiment 2, embryos were cultured in high or low oxygen tension with HTF:BME + Sigma antioxidant supplement® or in HTF:BME alone (Control). The data were analyzed using ANOVA followed by Tukey’s test. The results of experiment 1 showed a negative effect (P < 0.05) of vitamin E on blastocyst production in Group 2 (19.7 ± 0.1%). This effect was reduced in Group 3 by the addition of vitamin C (26.1 ± 0.2%). The use of vitamin C alone (34.9 ± 0.3%) or the Sigma antioxidant supplement® (33.3 ± 0.7%) did not increase (P > 0.05) the number of blastocysts produced compared with the control group (30.1 ± 0.5%). During experiment 2, there was no effect (P > 0.05) from the culture medium or the O2 concentrations used, indicating that the reduction of the O2 concentration did not improve blastocyst production.

Efeito do fator de crescimento epidermal (EGF) na maturação in vitro de oócitos caninos

Fundação de Apoio 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.

Avaliação dos fatores de virulência, atividade antimicrobiana e viabilidade celular de bactéria cariogênica na presença do Terpinen-4-ol: estudo in vitro

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação dos fatores de virulência, atividade antimicrobiana e viabilidade celular de bactéria cariogênica na presença do Terpinen-4-ol: estudo in vitro

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Micropropagation of gerbera using chlorine dioxide (ClO2) to sterilize the culture medium

One of the alternatives to autoclaving culture media is chemical sterilization, which may cause fewer changes to the chemical composition of the media. In this study, the effect of chemical sterilization by inclusion of chlorine dioxide (ClO2) in the culture medium on the in vitro development of gerbera (Gerbera jamesonii) cv. AL101, cultured at different stages of micropropagation, was evaluated. The following five concentrations of ClO2 were tested: 0%, 0.0025%, 0.0050%, 0.0075%, and 0.010%. Autoclaved medium was used as the control. ClO2 in the culture medium reduced contamination at rates comparable to autoclaving when tested at three stages of the culture process: in vitro establishment, multiplication, and rooting. Plantlets grown in culture media sterilized with ClO2 showed similar or better development than those grown in autoclaved culture medium. Use of 0.0025% ClO2 to sterilize the culture medium resulted in better plantlet development than autoclaved medium, regardless of the stage of micropropagation.

Bacterial leakage along the implant-abutment interface: culture and DNA Checkerboard hybridization analyses

Objective Bacterial species have been found harboring the internal surface of dental implants as consequence of their failed connections. The aim of the present study was to compare the detection frequency of bacterial leakage from human saliva through the implantabutment interface, under non-loading conditions, using either DNA Checkerboard or culture method. Materials and methods Thirty dental implants with hexagonal platforms were connected to pre-machined abutments according to the manufacturers specifications. The assemblies were individually incubated in human saliva under anaerobic conditions for 7 similar to days at 37 degrees C. Afterward, contents from the inner parts of the implants were collected and evaluated with either DNA Checkerboard (s similar to=similar to 15) or culture (n similar to=similar to 15). Subsequently, identification and quantitation of bacterial species from saliva and implants were carried out for the group evaluated with the DNA Checkerboard method. Results Both DNA Checkerboard and culture showed positive signals of bacterial leakage in 6 of the 15 evaluated samples. Capnocytophaga gingivalis and Streptococcus mutans were the most frequently detected species harboring the internal surface of the implants followed by Veillonella parvula. Conclusion Occurrence of bacterial leakage along the implantabutment interface is comparably detected with both DNA Checkerboard hybridization and conventional culture methods.

Characterization and in vitro evaluation of bacterial cellulose membranes functionalized with osteogenic growth peptide for bone tissue engineering

The aim of this study was to characterize the physicochemical properties of bacterial cellulose (BC) membranes functionalized with osteogenic growth peptide (OGP) and its C-terminal pentapeptide OGP[10-14], and to evaluate in vitro osteoinductive potential in early osteogenesis, besides, to evaluate cytotoxic, genotoxic and/or mutagenic effects. Peptide incorporation into the BC membranes did not change the morphology of BC nanofibers and BC crystallinity pattern. The characterization was complemented by Raman scattering, swelling ratio and mechanical tests. In vitro assays demonstrated no cytotoxic, genotoxic or mutagenic effects for any of the studied BC membranes. Culture with osteogenic cells revealed no difference in cell morphology among all the membranes tested. Cell viability/proliferation, total protein content, alkaline phosphatase activity and mineralization assays indicated that BC-OGP membranes enabled the highest development of the osteoblastic phenotype in vitro. In conclusion, the negative results of cytotoxicity, genotoxicity and mutagenicity indicated that all the membranes can be employed for medical supplies, mainly in bone tissue engineering/regeneration, due to their osteoinductive properties.

Histologic and functional evaluation of lungs reconditioned by ex vivo lung perfusion

BACKGROUND: Only about 15% of donor lungs are considered suitable for transplantation (LTx). Ex vivo lung perfusion (EVLP) has been developed as a method to reassess and repair damaged lungs. We report our experience with EVLP in non-acceptable donor lungs and evaluate its ability to recondition these lungs. METHODS: We studied lungs from 16 brain-dead donors rejected for LTx. After harvesting, the lungs were stored at 4 degrees C for 10 hours and subjected to normothermic EVLP with Steen Solution (Vitro life, Goteborg, Sweden) for 60 minutes. For functional evaluation, the following variables were assessed: partial pressure of arterial oxygen (Pao(2)), pulmonary vascular resistance (PVR), and lung compliance (LC). For histologic assessment, lung biopsy was done before harvest and after EVLP. Tissue samples were examined under light microscopy. To detect and quantify apoptosis, terminal deoxynucleotide transferase-mediated deoxy uridine triphosphate nick-end labeling assay was used. RESULTS: Thirteen lima donors were refused for having impaired lung function. The mean Pao(2) obtained in the organ donor at the referring hospital was 193.7 mm Hg and rose to 489 mm Hg after EVLP. During EVLP, the mean PVR was 652.5 dynes/sec/cm(5) and the mean LC was 48 ml/cm H2O. There was no significant difference between the mean Lung Injury Score before harvest and after EVLP. There was a trend toward a reduction in the median number of apoptotic cells after EVLP. CONCLUSIONS: EVLP improved lung function (oxygenation capacity) of organs considered unsuitable for transplantation. Lung tissue structure did not deteriorate even after 1 hour of normothermic perfusion. J Heart Lung Transplant 2012;31:305-9 (C) 2012 International Society for Heart and Lung Transplantation. All rights reserved.