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111.
112.
Cleoni Alves Mendes de Lima Harrison Magdinier Gomes Maraníbia Aparecida Cardoso Oelemann Jesus Pais Ramos Paulo Cezar Caldas Carlos Eduardo Dias Campos Márcia Aparecida da Silva Pereira Fátima Fandinho Onofre Montes Maria do Socorro Calixto de Oliveira Philip Noel Suffys Maria Manuela da Fonseca Moura 《Memórias do Instituto Oswaldo Cruz》2013,108(4):457-462
The main cause of pulmonary tuberculosis (TB) is infection with
Mycobacterium tuberculosis (MTB). We aimed to evaluate the
contribution of nontuberculous mycobacteria (NTM) to pulmonary disease in
patients from the state of Rondônia using respiratory samples and
epidemiological data from TB cases. Mycobacterium isolates were identified using
a combination of conventional tests, polymerase chain reaction-based restriction
enzyme analysis of hsp65 gene and hsp65 gene
sequencing. Among the 1,812 cases suspected of having pulmonary TB, 444 yielded
bacterial cultures, including 369 cases positive for MTB and 75 cases positive
for NTM. Within the latter group, 14 species were identified as
Mycobacterium abscessus, Mycobacterium
avium, Mycobacterium fortuitum,
Mycobacterium intracellulare, Mycobacterium
gilvum, Mycobacterium gordonae,
Mycobacterium asiaticum, Mycobacterium
tusciae, Mycobacterium porcinum,
Mycobacterium novocastrense, Mycobacterium
simiae, Mycobacterium szulgai,
Mycobacterium phlei and Mycobacterium
holsaticum and 13 isolates could not be identified at the species
level. The majority of NTM cases were observed in Porto Velho and the relative
frequency of NTM compared with MTB was highest in Ji-Paraná. In approximately
half of the TB subjects with NTM, a second sample containing NTM was obtained,
confirming this as the disease-causing agent. The most frequently observed NTM
species were M. abscessus and M. avium and
because the former species is resistant to many antibiotics and displays
unsatisfactory cure rates, the implementation of rapid identification of
mycobacterium species is of considerable importance. 相似文献
113.
João Vidigal Mafalda M. Dias Fabiana Fernandes Marco Patrone Cláudia Bispo Cláudia Andrade Rui Gardner Manuel J.T. Carrondo Paula M. Alves Ana P. Teixeira 《Journal of biotechnology》2013
Insect cell lines such as Sf9 and High Five™ have been widely used to produce recombinant proteins mostly by the lytic baculovirus vector system. We have recently established an expression platform in Sf9 cells using a fluorescence-based recombinase mediated cassette exchange (RMCE) strategy which has similar development timelines but avoids baculovirus infection. To expedite cell engineering efforts, a robust fluorescence-activated cell sorting (FACS) protocol optimized for insect cells was developed here. The standard sorting conditions used for mammalian cells proved to be unsuitable, resulting in post-sorting viabilities below 10% for both cell lines. We found that the extreme sensitivity to the shear stress displayed by Sf9 and High Five™ cells was the limiting factor, and using Pluronic F-68 in the cell suspension could increase post-sorting viabilities in a dose dependent manner. The newly developed protocol was then used to sort stable populations of both cell lines tagged with a DsRed-expressing cassette. Before sorting, the average fluorescence intensity of the Sf9 cell population was 3-fold higher than that of the High Five™ cell population. By enriching with the 10% strongest DsRed-fluorescent cells, the productivity of both cell populations could be successfully improved. The established sorting protocol potentiates the use of RMCE technology for recombinant protein production in insect cells. 相似文献
114.
Alexandre G. Rodrigues Liu Yu Ping Priscyla D. Marcato Oswaldo L. Alves Maria C. P. Silva Rita C. Ruiz Itamar S. Melo Ljubica Tasic Ana O. De Souza 《Applied microbiology and biotechnology》2013,97(2):775-782
Aspergillus tubingensis and Bionectria ochroleuca showed excellent extracellular ability to synthesize silver nanoparticles (Ag NP), spherical in shape and 35?±?10 nm in size. Ag NP were characterized by transmission electron microscopy, X-ray diffraction analysis, and photon correlation spectroscopy for particle size and zeta potential. Proteins present in the fungal filtrate and in Ag NP dispersion were analyzed by electrophoresis (sodium dodecyl sulfate polyacrylamide gel electrophoresis). Ag NP showed pronounced antifungal activity against Candida sp, frequently occurring in hospital infections, with minimal inhibitory concentration in the range of 0.11–1.75 μg/mL. Regarding antibacterial activity, nanoparticles produced by A. tubingensis were more effective compared to the other fungus, inhibiting 98.0 % of Pseudomonas. aeruginosa growth at 0.28 μg/mL. A. tubingensis synthesized Ag NP with surprisingly high and positive surface potential, differing greatly from all known fungi. These data open the possibility of obtaining biogenic Ag NP with positive surface potential and new applications. 相似文献
115.
Camila Arantes Hartmann Vilma Regina Martins Flavia Regina Souza Lima 《FEBS letters》2013,587(2):238-244
Prion protein (PrPC) has neuroprotective functions and herein we demonstrate that astrocytes from PrPC-over-expressing mice are more resistant to induced cell death than wild-type astrocytes. The Stress-Inducible-Protein 1 (STI1), a PrPC ligand, prevents cell death in both wild-type and PrPC-over-expressing astrocytes through the activation of protein-kinase-A. Cultured embryonic astrocytes and brain extracts from PrPC-over-expressing mice show higher glial fibrillary acidic protein expression and reduced vimentin and nestin levels when compared to wild-type astrocytes, suggesting faster astrocyte maturation in the former mice. Our data indicate that PrPC levels modulate astrocyte development, and that PrPC–STI1 interaction contributes to protect against astrocyte death. 相似文献
116.
117.
Lucas Cunha Dias de Rezende Fernando Fumagalli Marraiana Schiavon Bortolin Marianne Garcia de Oliveira Murilo Helder de Paula Valter Ferreira de Andrade-Neto Flavio da Silva Emery 《Bioorganic & medicinal chemistry letters》2013,23(16):4583-4586
1,4-Naphthoquinone derivatives are known to have relevant activities against several parasites. Among the treatment options for malaria, atovaquone, a 1,4-naphthoquinone derivative, is widely applied in the treatment and prophylaxis of such disease. Based on the structure simplification of atovaquone, we designed and synthesized four novel naphthoquinoidal derivatives. The compounds were obtained by the underexplored epoxide-opening reaction of 1,4-naphthoquinone using aniline derivatives as nucleophiles. The antiplasmodial activity of the synthesized compounds was performed in vivo using Peter’s 4 days suppression test. Significant parasitemia reduction and increased survival were observed for some of the compounds. 相似文献
118.
Daniela Volcan Almeida Camila de Martinez Gaspar Martins M��rcio de Azevedo Figueiredo Carlos Frederico Ceccon Lanes Adalto Bianchini Luis Fernando Marins 《Transgenic research》2013,22(1):75-88
Growth hormone (GH) transgenic fish are at a critical step for possible approval for commercialization. Since this hormone is related to salinity tolerance in fish, our main goal was to verify whether the osmoregulatory capacity of the stenohaline zebrafish (Danio rerio) would be modified by GH-transgenesis. For this, we transferred GH-transgenic zebrafish (T) from freshwater to 11 ppt salinity and analyzed survival as well as relative changes in gene expression. Results show an increased mortality in T versus non-transgenic (NT) fish, suggesting an impaired mechanism of osmotic acclimation in T. The salinity effect on expression of genes related to osmoregulation, the somatotropic axis and energy metabolism was evaluated in gills and liver of T and NT. Genes coding for Na+, K+-ATPase, H+-ATPase, plasma carbonic anhydrase and cytosolic carbonic anhydrase were up-regulated in gills of transgenics in freshwater. The growth hormone receptor gene was down-regulated in gills and liver of both NT and T exposed to 11 ppt salinity, while insulin-like growth factor-1 was down-regulated in liver of NT and in gills of T exposed to 11 ppt salinity. In transgenics, all osmoregulation-related genes and the citrate synthase gene were down-regulated in gills of fish exposed to 11 ppt salinity, while lactate dehydrogenase expression was up-regulated in liver. Na+, K+-ATPase activity was higher in gills of T exposed to 11 ppt salinity as well as the whole body content of Na+. Increased ATP content was observed in gills of both NT and T exposed to 11 ppt salinity, being statistically higher in T than NT. Taking altogether, these findings support the hypothesis that GH-transgenesis increases Na+ import capacity and energetic demand, promoting an unfavorable osmotic and energetic physiological status and making this transgenic fish intolerant of hyperosmotic environments. 相似文献
119.
Tatiane Sanches Soares Ricardo Jose Soares Torquato Francisco Jose Alves Lemos Aparecida Sadae Tanaka 《Insect biochemistry and molecular biology》2013,43(1):9-16
Dengue is a serious disease transmitted by the mosquito Aedes aegypti during blood meal feeding. It is estimated that the dengue virus is transmitted to millions of individuals each year in tropical and subtropical areas. Dengue control strategies have been based on controlling the vector, Ae. aegypti, using insecticide, but the emergence of resistance poses new challenges. The aim of this study was the identification of specific protease inhibitors of the digestive enzymes from Ae. aegypti larvae, which may serve as a prospective alternative biocontrol method. High affinity protein inhibitors were selected by all of the digestive serine proteases of the 4th instar larval midgut, and the specificity of these inhibitors was characterized. These inhibitors were obtained from a phage library displaying variants of HiTI, a trypsin inhibitor from Haematobia irritans, that are mutated in the reactive loop (P1–P4′). Based on the selected amino acid sequence pattern, seven HiTI inhibitor variants were cloned, expressed and purified. The results indicate that the HiTI variants named T6 (RGGAV) and T128 (WNEGL) were selected by larval trypsin-like (IC50 of 1.1 nM) and chymotrypsin-like enzymes (IC50 of 11.6 nM), respectively. The variants T23 (LLGGL) and T149 (GGVWR) inhibited both larval chymotrypsin-like (IC50 of 4.2 nM and 29.0 nM, respectively) and elastase-like enzymes (IC50 of 1.2 nM for both). Specific inhibitors were successfully obtained for the digestive enzymes of Ae. aegypti larvae by phage display. Our data also strongly suggest the presence of elastase-like enzymes in Ae. aegypti larvae. The HiTI variants T6 and T23 are good candidates for the development as a larvicide to control the vector. 相似文献
120.
Claudia Mara Maciel-Rezende Letícia de Almeida Éderson D’Martin Costa Francieli Ribeiro Pires Karina Ferreira Alves Cláudio Viegas Junior Danielle Ferreira Dias Antônio Carlos Doriguetto Marcos José Marques Marcelo Henrique dos Santos 《Bioorganic & medicinal chemistry》2013,21(11):3114-3119
Nine O-alkyl and O-prenyl derivatives were synthesized from commercial 2,4-dihydroxybenzophenone, 4,e4,4′-dihydroxybenzophenone and were evaluated for their leishmanicidal activity against promastigote forms of Leishmania amazonensis, as well their toxicity in murine macrophages. All derivatives exhibited better biological activity than their hydroxylated benzophenones precursors, and new compound LFQM-123 (3c) was 250-fold more active than its precursor 4,4′-dihydroxybenzophenone (3). Moreover, some of the results were comparable to the standard drug Amphotericin B, suggesting that the increase in lipophilicity could facilitate protozoa membrane permeation. In this study we confirmed that benzophenone derivatives exhibit leishmanicidal properties, with relatively low toxicity, and thus could be exploited as promise prototypes for the design and development of new drug for the treatment of leishmaniasis. 相似文献