Obtenção de mutantes de Bacillus thuringiensis por inserção do transposon Tn-5 e avaliação da toxicidade frente a larvas de Spodoptera frugiperda

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

Pontas e velocidade de deslocamento na deposição de gotas da pulverização na cultura do algodão

Pós-graduação em Agronomia (Proteção de Plantas) - FCA

SNPs (Single Nucleotide Polymorphisms) em genes relacionados a absorção e translocação de Boro em Eucalyptus

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Entrapment characteristics of hydrosoluble vitamins loaded into chitosan and N,N,N-trimethyl chitosan nanoparticles

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

Utilização da percepção subjetiva de esforço para controlar a intensidade do aquecimento realizado através da corrida em crianças

The aim of this study was to verify the use of exertion subjective perception to control warm-up intensity performed by continuous running in children. Twenty-one children (11,4 ± 0,8 years, 45 ± 9,8 kg e 153,6 ± 9,6 cm), performed warm-up by continuous running in randomic order in three conditions with diferent intensities using rate of perceived exertion (RPE). The intensities were chosen according to RPE Borg CR-10 and the arbitrary units used were three (B3), Five (B5) and nine (B9). The heart rate (HR) and mean velocity of the run (VEL) were evaluated to analyse the intensity of the three warm-up conditions. Statistical analysis was done by annova one-way followed by post-hoc tukey test, considering significant level of 5%. There were significant increase in HR (B3= 112 ± 10, B5 = 140 ± 18 e B9 = 176 ± 21 bpm) and VEL (B3 = 4,01 ± 0,5, B5 = 7,09 ± 1,22 e B9 = 10,97 ± 1,70 km/h) between B5 and B9 condition compared to B3. Moreover, in both parameters B9 presented significantly higher results compared to the condition B5. It´s possible to control warm-up intensity performed by continuous running in children using RPE Borg CR-10.

Genus Bifidobacterium: taxonomy studies and gene expression analysis on folate pathway

Folates (vitamin B9) are essential water soluble vitamins, whose deficiency in humans may contribute to the onset of several diseases, such as anaemia, cancer, cardiovascular diseases, neurological problems as well as defects in embryonic development. Human and other mammals are unable to synthesize ex novo folate obtaining it from exogenous sources, via intestinal absorption. Recently the gut microbiota has been identified as an important source of folates and the selection and use of folate producing microorganisms represents an innovative strategy to increase human folate levels. The aim of this thesis was to gain a fundamental understanding of folate metabolism in Bifidobacterium adolescentis. The work was subdivided in three main phases, also aimed to solve different problems encountered working with Bifidobacterium strains. First, a new identification method (based on PCR-RFLP of hsp60 gene) was specifically developed to identify Bifidobacterium strains. Secondly, Bifidobacterium adolescentis biodiversity was explored in order to recognize representing strains of this species to be screened for their folate production ability. Results showed that this species is characterized by a wide variability and support the idea that a possible new taxonomic re-organization would be required. Finally B. adolescentis folate metabolism was studied using a double approach. A quantitative analysis of folate content was complemented by the examination of expression levels of genes involved in folate related pathways. For the normalization process, required to increase the robustness of the qRT-PCR analysis, an appropriate set of reference genes was tested using two different algorithms. Results demonstrate that B.adolescentis strains may represent an endogenous source of natural folate and they could be used to fortify fermented dairy products. This bio-fortification strategy presents many advantages for the consumer, providing native folate forms more bio-available, and not implicated in the discussed controversy concerning the safety of high intake of synthetic folic acid.

Folic acid derivatives for PET imaging and therapy addressing folate receptor positive tumors

Folic acid, also known as vitamin B9, is the oxidized form of 5,6,7,8-tetrahydrofolate, which serves as methyl- or methylene donor (C1-building blocks) during DNA synthesis. Under physiological conditions the required amount of 5,6,7,8-tetrahydrofolate for survival of the cell is accomplished through the reduced folate carrier (RFC). In contrast, the supply of 5,6,7,8-tetrahydrofolate is insufficient under pathophysiological conditions of tumors due to an increased proliferation rate. Consequently, many tumor cells exhibit an (over)expression of the folate receptor. This phenomenon has been applied to diagnostics (PET, SPECT, MR) to image FR-positive tumors and on the other hand to treat malignancies related to a FR (over)expression. Based on this concept, a new 18F-labeled folate for PET imaging has been developed and was evaluated in vivo using tumor-bearing mice. The incorporation of oligoethylene spacers into the molecular structure led to a significant enhancement of the pharmacokinetics in comparison to previously developed 18F-folates. The liver uptake could be reduced by one sixth by remaining a tumor uptake of 3%ID/g leading to better contrast ratios. Encouraged by these results, a clickable 18F-labeled serine-based prosthetic group has been synthesized, again with the idea to improve the metabolic and pharmacokinetic profile of hydrophilic radiotracers. Therefore, an alkyne-carrying azido-functionalized serine derivative for coupling to biomolecules was synthesized and a chlorine leaving group for 18F-labeling, which could be accomplished using a microwave-assisted synthesis, a [K⊂2.2.2]+/carbonate system in DMSO. Radiochemical yields of 77±6% could be achieved.rnThe promising results obtained from the FR-targeting concept in the diagnostic field have been transferred to the boron neutron capture therapy. Therefore, a folate derivative was coupled to different boron clusters and cell uptake studies were conducted. The synthesis of the folate-boron clusters was straightforward. At first, a linker molecule based on maleic acid was synthesized, which was coupled to the boron cluster via Michael Addition of a thiol and alkene and subsequently coupled to the targeting moiety using CuAAC. The new conjugates of folate and boron clusters led to a significant increase of boron concentration in the cell of about 5-times compared to currently used and approved boron pharmaceuticals. rnMoreover, azido-folate derivatives were coupled to macromolecular carrier systems (pHPMA), which showed an enhanced and specific accumulation at target sites (up to 2.5-times) during in vivo experiments. A specific blockade could be observed up to 30% indicating an efficient targeting effect. A new kind of nanoparticles consisting of a PDLLA core and p((HPMA)-b-LMA)) as surfactants were developed and successfully radiolabeled via 18F-click chemistry in good RCYs of 8±3%rnThe nanoparticles were obtained via the miniemulsion technique in combination with solvent evaporation. The 18F-labeled nanoparticles were applied to in vivo testing using a mouse model. PET imaging showed a “mixed” biodistribution of low molecular weight as well as high molecular weight systems, indicating a partial loss of the 18F-labeled surfactant.rnIn conclusion, the presented work successfully utilized the FR-targeting concept in both, the diagnostic field (PET imaging) and for therapeutic approaches (BNCT, drug delivery systems). As a result, the high potential of FR-targeting in oncological applications has been shown and was confirmed by small animal PET imaging.rn

Subarachnoid hemorrhage and intracerebral hematoma: incidence, prognostic factors, and outcome

OBJECTIVE: To analyze the incidence and impact of an intracerebral hematoma (ICH) on treatment and outcome in patients with aneurysmal subarachnoid hemorrhage. METHODS: Data of 585 consecutive patients with subarachnoid hemorrhage from June 1999 to December 2005 were prospectively entered in a database. ICH was diagnosed and size was measured by computed tomographic scan before aneurysm occlusion. Fifty patients (8.5%) presented with an ICH larger than 50 cm3. The treatment decision (coil, clip, or hematoma evacuation) was based on an interdisciplinary approach. Patients were stratified into good (Hunt and Hess Grades I-III) versus poor (Hunt and Hess Grades IV and V) grade, and outcome was assessed according to the modified Rankin Scale at 6 months. RESULTS: Overall, 358 patients presented in good grade, with 4 of them having ICH (1.1%); and 227 patients presented in poor grade, with 46 of them having ICH (20.3%, P < 0.01). In good-grade patients with an ICH (n = 4), a favorable outcome (modified Rankin Scale score of 0-2) was achieved in 1 patient (25%), and in 246 patients (75%) without an ICH (P = 0.053; odds ratio, 0.11). A favorable outcome was achieved in 5 poor-grade patients (12.8%) with an ICH and in 40 patients (23.7%) without an ICH (P = 0.19; odds ratio, 0.47). Time to treatment was significantly shorter in patients with an ICH than without an ICH (median, 7 versus 26 h; P < 0.001) and shortest in patients with favorable outcome (3.5 hours; P < 0.01). CONCLUSION: The current data confirm that the presence of an ICH is a predictor of unfavorable outcome. However, despite large ICHs, a significant number of patients have a good outcome. To achieve a favorable outcome, ultra-early treatment with hematoma evacuation and aneurysm obliteration seems to be mandatory.

Crustal uplift rates of 4 GPS stations near Jakobshavn Isbrae (2006-2009)

We analyze 2006-2009 data from four continuous Global Positioning System (GPS) receivers located between 5 and 150 km from the glacier Jakobshavn Isbrae, West Greenland. The GPS stations were established on bedrock to determine the vertical crustal motion due to the unloading of ice from Jakobshavn Isbrae. All stations experienced uplift, but the uplift rate at Kangia North, only 5 km from the glacier front, was about 10 mm/yr larger than the rate at Ilulissat, located only ~45 km further away. This suggests that most of the uplift is due to the unloading of the Earth's surface as Jakobshavn thins and loses mass. Our estimate of Jakobshavn's contribution to uplift rates at Kangia North and Ilulissat are 14.6 ± 1.7 mm/yr and 4.9 ± 1.1 mm/yr, respectively. The observed rates are consistent with a glacier thinning model based on repeat altimeter surveys from NASA's Airborne Topographic Mapper (ATM), which shows that Jakobshavn lost mass at an average rate of 22 ± 2 km**3/yr between 2006 and 2009. At Kangia North and Ilulissat, the predicted uplift rates computed using thinning estimates from the ATM laser altimetry are 12.1 ± 0.9 mm/yr and 3.2 ± 0.3 mm/yr, respectively. The observed rates are slightly larger than the predicted rates. The fact that the GPS uplift rates are much larger closer to Jakobshavn than further away, and are consistent with rates inferred using the ATM-based glacier thinning model, shows that GPS measurements of crustal motion are a potentially useful method for assessing ice-mass change models.

Coccoliths in phytoplankton of the Eastern Bering Sea in August 2001

Based on results of field observations in August 1998, July 2000, and August 2001 composition and quantitative distribution of coccolithophorids in the middle part of the Eastern Bering Sea shelf between 56°052'N and 59°019'N was characterized. Emiliania huxleyi abundance, biomass, and population structure as well as role of species in the coccolithophorid community and phytoplankton as a whole were evaluated. Abundance of the species in the upper mixed layer in bloom areas was 1-3 mln cells/l and biomass made up 30-75 mg C/m**3. E. huxleyi share in total phytoplankton numbers and biomass at that reached 98% and 84% respectively. Significant spatial heterogeneity of E. huxleyi, quantitative distribution and population size structure, as well as asynchronism in population development in neighboring parts of the bloom area were shown. The time period, during which population structure in certain part of the area shifts from domination of juvenile cells without coccoliths to a phase of active detritus formation with dying coccolithophorid cells involved, may be estimated as two weeks. A conclusion is made that after anomalous E. huxleyi bloom in 1997 mass development of coccolithophorids became a characteristic feature of phytoplankton community's seasonal succession in the middle part of the Eastern Bering Sea shelf.