Cyclopentac]thiophene oligomers based solution processable D-A copolymers and their application as FET materials

Two new solution processable, low band gap donor-acceptor (D-A) copolymers (P1 and P2) comprising a cyclopentac] thiophene (CPT) based oligomers as donors and benzoc]1,2,5] selenadiazole (BDS) and 2-dodecyl1,2,3]-benzotriazole (BTAz) as acceptors were synthesized and characterized and their field effect transistor properties were studied. The internal charge transfer interaction between the electron-donating CPT based oligothiophene and the electron-accepting BDS or BTAz unit effectively reduces the band gap in polymers to 1.3 and 1.66 eV with low lying highest occupied molecular orbital (HOMO). The absorption spectrum of P1 was found to be more red shifted than that of P2 because of incorporation of the more electron-withdrawing BDS unit. The color of neutral P1 was found to be green in both solution and film states with two major bands in the absorption spectra; however, neutral P2 revealed one dominant absorption exhibiting red color in both solution and film state which could be attributed to the less electron-withdrawing effect of the BTAz unit. The polymers were further characterized by GPC, TGA, DSC and cyclic voltammetry. P1 and P2 exhibited charge carrier mobilities as high as 9 x 10(-3) cm(2) V-1 s(-1) and 2.56 x 10(-3) cm 2 V-1 s(-1), respectively with the current on/off ratio (I-on/I-off) in the order of 10(2).

A variant peptide of buffalo colostrum beta-lactoglobulin inhibits angiotensin I-converting enzyme activity

beta-lactoglobulin is a rich source of bioactive peptides. The LC-MS separated tryptic peptides of buffalo colostrum beta-lactoglobulin (BLG-col) were computed based on MS-MS fragmentation for de novo sequencing. Among the selected peptides (P1-P8), a variant was detected with methionine at position 74 instead of glutamate. The sequences of two peptides were identical to hypocholesterolemic peptides whereas the remaining peptides were in accordance with buffalo milk beta-lactoglobulin. Comparative sequence analysis of BLG-col to milk beta-lactoglobulin was carried out using CLUSTALW2 and a molecular model for BLG-col was constructed (PMDB ID-PM0076812). The synthesized variant pentapeptide (IIAMK, m/z-576 Da) was found to inhibit angiotensin I-converting enzyme (ACE) with an IC50 of 498 +/- 2 mu M, which was rationalized through docking simulations using Molgrow virtual docker. (C) 2012 Elsevier Masson SAS. All rights reserved.

Temperature measurement of reflected shock wave by using chemical indicator

This report describes a new method for measuring the temperature of the gas behind the reflected shock wave in shock tube, corresponding to the reservoir temperature of a shock tunnel, based on the chemical reaction of small amount of CF4 premixed in the test gas. The final product C2F4 is used as the temperature indicator, which is sampled and detected by a gas chromatography in the experiment. The detected concentration of C2F4 is correlated to the temperature of the reflected shock wave with the initial pressure P-1 and test time tau as parameters in the temperature range 3 300 K < T < 5 600 K, pressure range 5 kPa < P1 <12 kPa and tau similar or equal to 0.4 ms.

Influencia de diferentes periodos de destete en conejas primiparas en el municipio de Diriamba, Departamento de Carazo, Nicaragua

Este trabajo fue realizado en la Quinta "Myriam", ubicada en el municipio de Diriamba, departamento de Carazo, Nicaragua, propiedad del Dr. Jorge Ferreira. La ubicación geográfica del municipio está comprendida entre los 11° 50' y 12° 00' de Latitud Norte y entre los 86° 00' y 86° 15' de Longitud Oeste, cuenta con una elevación de 551 m.s.n.m., los datos meteorológicos de la zona en los últimos cuatro años (90-93) en cuanto a temperatura, humedad relativa y precipitación promedio fueron: 23.8 °C, 84% y 1184,5 mm respectivamente, (INETER, Departamento de Estadística, 1993). En el presente ensayo, se utilizó un diseño estadístico completamente aleatorizado (DCA), mediante el cual se evaluó el efecto de 3 tratamientos (15, 30 y 45 días de destete) sobre las variables Peso de la Camada al Destete (PCD), Peso de la Madre al Destete (PMD), Peso de la Madre al Segundo Parto (PSP), Ganancia Media Diaria del Parto 1 al Parto 2 (GMDp 1.P2) , con 4 repeticiones por tratamiento, éstas repeticiones estuvieron constituidas por conejas primíparas, las cuales tuvieron un peso que osciló entre 3 - 3.18 kg. y edad homogénea de 7 meses. Las variables fueron sometidas a la prueba de Duncan para determinar la superioridad en rango por tratamiento. Mediante el análisis de varianza se encontró que el efecto de los tratamientos resultó ser altamente significativo sobre la variable Peso de la Camada al Destete (PCD) (P0.01), significativo sobre las variables Peso de la Madre al Segundo Parto (PSP) y Ganancia Media Diaria del Parto 1 al Parto 2 (GMD p 1.P2) (P0.1) y no significativo sobre la variable Peso de la Madre al Destete (PMD) (NS). El efecto de la covariable Peso al Primer Parto (P1P) resultó ser altamente significativo sobre las variables Peso de la Madre al Destete (PMD) y Peso de la Madre al Segundo Parto (PSP) (P0.01). Después de someter las variables a la prueba de DUNCAN se encontraron los siguientes promedios por tratamiento: Peso de la Camada al Destete (PCD) ; T.15 días 1.538 kg P.V.(C), T.30 días 3.360 kg P.V. (B), T.45 días 4.523 kg P.V. (A), Peso de la Madre al Destete (PMD); T.15 días 3.649 kg P.V. (A) T.30 días 3.549 kg P.V. (A), T.45 dias 3.493 kg P.V.(A). Peso de la Madre al Segundo Parto (PSP); T.15 días 3. 449 kg P.V. (B), T.30 días 3.628 kg P.V. (AB), T.45 días 3.777 kg P.V.(A). Ganancia Media Diaria del Parto 1 al Parto 2 (GMD p 1.P2) T .15 días (2.071 g (B), T.30 días 2.718 g (AB), T.45 días 2. 975 g (A). Los promedios y desviaciones standard para las variables Ganancia Media Diaria del Parto 1 al Destete 1 (GMDp1-D1)y Ganancia Media Diaria del Destete l al Parto 2 (GMDD1.P2) por tratamiento fueron: GMD P1-D1; T.l5 días 3.84 g ± 0.50 g, T.30 días 2.54 g ± 0.64 g, T.45 días 2.37 g ± 0.72 g. GMD D1-P2; T.l5 días 2.26 g ± 0.54 g, T.30 días 3.17 g ± 0.74 g, T.45 días 3.93 g ± 0.28 g. Se encontró que el alimento tipo único suministrado a los animales bajo experimentación, cumplió con los requerimientos nutricionales de la especie estudiada. Los costos encontrados por hembra, por tratamiento, fueron C$ 197.21 destete de 15 días, C$ 220.28 destete de 30 días, C$ 243 .1.4 destete de 45 días. Los costos por gazapo destetado fueron de C$ 5.48 destete de 15 días, C$ 6.12 destete de 30 días, C$ 8.10 destete de 45 días.

强激波阵面的非平衡结构研究

利用测量强激波波后N2+第一负系(0,0)带和(1,2)带的辐射,对强激波后振动温度历程的测量过程进行了探索,并利用Langmuir探针技术,在低密度激波管中对强激波后电子数密度历程进行了测量.测量和计算结果进行了对比.结果表明:N2+B2∑u+态的激发比振动能的激发更快;实验测得的振动温度有明显的周期性振荡;在激波速度7.65～7.85km/s、p1=1.33Pa、实验段内径0.8m下,实验有效时间只有约6.5μs,实验中的电子数密度不能达到峰值.在约10倍波前自由程的实验有效区域内,电子数密度的测量值与计算值吻合很好.

Análisis de la estructura varisca y alpina en la transversal Sallent-Biescas (Pirineos Centrales, Huesca)

186 p.

Iso-specific Ziegler-Natta polymerization of α-olefins with a single-component organoyttrium catalyst

The synthesis of a sterically tailored ligand array (M)_2((C_5H_2-2-Si(CH_3)_3-4-C(CH_3)_3)S_2i(CH_3)_2]("M_2Bp") (M = Li, 16; K, 19) is described. Transmetallation of Li_2Bp with YCl_3(THF)_3 affords exclusively the C_2 symmetric product rac-[BpY(µ_2-Cl)_2Li(THF)_2], 20. A X-ray crystal structure of 20 has been determined; triclinic, P1, a= 13.110 (8), b = 17.163 (15), c = 20.623 (14) Å, α = 104.02 (7), β = 99.38 (5), γ = 100.24 (6)° , Z = 4, R = 0.056. Transmetallation of K_2Bp with YCl_3(THF)_3 affords the halide free complex rac-BpYCl, 23. The corresponding rac-BpLaCl, 28, is prepared in an anlogous manner. In all cases the achiral meso isomer is not obtained since only for the racemic isomers are the unfavorable steric interactions between the Si(CH3)_3 groups in the narrow portion of the [Cp-M'-Cp] wedge avoided. Alkylation of 20 or 23 with LiCH(Si(CH_3)_3)_2 affords rac-BpYCH(Si(CH_3)_3)_2, 26 in good yield. Alkylation of 28 with LiCH(Si(CH_3)_3)_2 affords rac-BpLaCH(Si(CH_3)_3)_2 29. Hydrogenation of 26 cleanly affords the bridging hydride species [BpY(µ_2-H)]_2, 27, as the homochiral (R,R) and (S,S) dimeric pairs. 26 is an efficient initiator for the polymerization of ethylene to high molecular weight linear polyethylene. 27 catalyzes the polymerization of propylene (25% v/v in methylcyclohexane) and neat samples of 1-butene, 1-pentene, 1-hexene to moderately high molecular weight polymers: polypropylene (M_n = 4,200, PDI 2.32, T_m 157 °C); poly-1-butene (M_n = 8,500, PDI 3.44, T_m 105 °C); poly-1-pentene (M_n = 20,000, PDI 1.99, T_m 73 °C); poly-1-hexene (M_n = 24,000, PDI 1.75, T_m < 25 °C). ^(13)C NMR spectra at the pentad analysis level indicates that the degree of isotacticity is 99% mmmm for all polymer samples. 27 is the first single component iso-specific α-olefin polymerization catalyst. The presumed origins of the high isospecificity are presented.

Structure-, stereochemistry-, and metal-regulated DNA binding/cleaving molecules

In the cell, the binding of proteins to specific sequences of double helical DNA is essential for controlling the processes of protein synthesis (at the level of DNA transcription) and cell proliferation (at the level of DNA replication). In the laboratory, the sequence-specific DNA binding/cleaving properties of restriction endonuclease enzymes (secreted by microorganisms to protect them from foreign DNA molecules) have helped to fuel a revolution in molecular biology. The strength and specificity of a protein:DNA interaction depend upon structural features inherent to the protein and DNA sequences, but it is now appreciated that these features (and therefore protein:DNA complexation) may be altered (regulated) by other protein:DNA complexes, or by environmental factors such as temperature or the presence of specific organic molecules or inorganic ions. It is also now appreciated that molecules much smaller than proteins (including antibiotics of molecular weight less than 2000 and oligonucleotides) can bind to double-helical DNA in sequence-specific fashion. Elucidation of structural motifs and microscopic interactions responsible for the specific molecular recognition of DNA leads to greater understanding of natural processes and provides a basis for the design of novel sequence-specific DNA binding molecules. This thesis describes the synthesis and DNA binding/cleaving characteristics of molecules designed to probe structural, stereochemical, and environmental factors that regulate sequence-specific DNA recognition.

Chapter One introduces the DNA minor groove binding antibiotics Netropsin and Distamycin A, which are di- and tri(N-methylpyrrolecarboxamide) peptides, respectively. The method of DNA affinity cleaving, which has been employed to determine DNA binding properties of designed synthetic molecules is described. The design and synthesis of a series of Netropsin dimers linked in tail-to-tail fashion (by oxalic, malonic, succinic, or fumaric acid), or in head-to-tail fashion (by glycine, β-alanine, and γ-aminobutanoic acid (Gaba)) are presented. These Bis(Netropsin)s were appended with the iron-chelating functionality EDTA in order to make use of the technique of DNA affinity cleaving. Bis(Netropsin)-EDTA compounds are analogs of penta(N-methylpyrrolecarboxamide)-EDTA (P5E), which may be considered a head-to-tail Netropsin dimer linked by Nmethylpyrrolecarboxamide. Low- and high-resolution analysis of pBR322 DNA affinity cleaving by the iron complexes of these molecules indicated that small changes in the length and nature of the linker had significant effects on DNA binding/cleaving efficiency (a measure of DNA binding affinity). DNA binding/cleaving efficiency was found to decrease with changes in the linker in the order β-alanine > succinamide > fumaramide > N-methylpyrrolecarboxamide > malonamide >glycine, γ-aminobutanamide > oxalamide. In general, the Bis(Netropsin)-EDTA:Fe compounds retained the specificity for seven contiguous A:T base pairs characteristic of P5E:Fe binding. However, Bis(Netropsin)Oxalamide- EDTA:Fe exhibited decreased specificity for A:T base pairs, and Bis(Netropsin)-Gaba-EDT A:Fe exhibited some DNA binding sites of less than seven base pairs. Bis(Netropsin)s linked with diacids have C₂-symmmetrical DNA binding subunits and exhibited little DNA binding orientation preference. Bis(Netropsin)s linked with amino acids lack C₂-symmetrical DNA binding subunits and exhibited higher orientation preferences. A model for the high DNA binding orientation preferences observed with head-to-tail DNA minor groove binding molecules is presented.

Chapter Two describes the design, synthesis, and DNA binding properties of a series of chiral molecules: Bis(Netropsin)-EDTA compounds with linkers derived from (R,R)-, (S,S)-, and (RS,SR)-tartaric acids, (R,R)-, (S,S)-, and (RS,SR)-tartaric acid acetonides, (R)- and (S)-malic acids, N ,N-dimethylaminoaspartic acid, and (R)- and (S)-alanine, as well as three constitutional isomers in which an N-methylpyrrolecarboxamide (P1) subunit and a tri(N-methylpyrrolecarboxamide)-EDTA (P3-EDTA) subunit were linked by succinic acid, (R ,R)-, and (S ,S)-tartaric acids. DNA binding/cleaving efficiencies among this series of molecules and the Bis(Netropsin)s described in Chapter One were found to decrease with changes in the linker in the order β-alanine > succinamide > P1-succinamide-P3 > fumaramide > (S)-malicamide > N-methylpyrrolecarboxamide > (R)-malicamide > malonamide > N ,N-dimethylaminoaspanamide > glycine = Gaba = (S,S)-tartaramide = P1-(S,S)-tanaramide-P3 > oxalamide > (RS,SR)-tartaramide = P1- (R,R)-tanaramide-P3 > (R,R)-tartaramide (no sequence-specific DNA binding was detected for Bis(Netropsin)s linked by (R)- or (S)-alanine or by tartaric acid acetonides). The chiral molecules retained DNA binding specificity for seven contiguous A:T base pairs. From the DNA affinity cleaving data it could be determined that: 1) Addition of one or two substituents to the linker of Bis(Netropsin)-Succinamide resulted in stepwise decreases in DNA binding affinity; 2) molecules with single hydroxyl substituents bound DNA more strongly than molecules with single dimethylamino substituents; 3) hydroxyl-substituted molecules of (S) configuration bound more strongly to DNA than molecules of (R) configuration. This stereochemical regulation of DNA binding is proposed to arise from the inherent right-handed twist of (S)-enantiomeric Bis(Netropsin)s versus the inherent lefthanded twist of (R)-enantiomeric Bis(Netropsin)s, which makes the (S)-enantiomers more complementary to the right-handed twist of B form DNA.

Chapter Three describes the design and synthesis of molecules for the study of metalloregulated DNA binding phenomena. Among a series of Bis(Netropsin)-EDTA compounds linked by homologous tethers bearing four, five, or six oxygen atoms, the Bis(Netropsin) linked by a pentaether tether exhibited strongly enhanced DNA binding/cleaving in the presence of strontium or barium cations. The observed metallospecificity was consistent with the known affinities of metal cations for the cyclic hexaether 18-crown-6 in water. High-resolution DNA affinity cleaving analysis indicated that DNA binding by this molecule in the presence of strontium or barium was not only stronger but of different sequence-specificity than the (weak) binding observed in the absence of metal cations. The metalloregulated binding sites were consistent with A:T binding by the Netropsin subunits and G:C binding by a strontium or barium:pentaether complex. A model for the observed positive metalloregulation and novel sequence-specificity is presented. The effects of 44 different cations on DNA affinity cleaving by P5E:Fe were examined. A series of Bis(Netropsin)-EDTA compounds linked by tethers bearing two, three, four, or five amino groups was also synthesized. These molecules exhibited strong and specific binding to A:T rich regions of DNA. It was found that the iron complexes of these molecules bound and cleaved DNA most efficiently at pH 6.0-6.5, while P5E:Fe bound and cleaved most efficiently at pH 7.5-8.0. Incubating the Bis(Netropsin) Polyamine-EDTA:Fe molecules with K₂PdCl₄ abolished their DNA binding/cleaving activity. It is proposed that the observed negative metalloregulation arises from kinetically inert Bis(Netropsin) Polyamine:Pd(II) complexes or aggregates, which are sterically unsuitable for DNA complexation. Finally, attempts to produce a synthetic metalloregulated DNA binding protein are described. For this study, five derivatives of a synthetic 52 amino acid residue DNA binding/cleaving protein were produced. The synthetic mutant proteins carried a novel pentaether ionophoric amino acid residue at different positions within the primary sequence. The proteins did not exhibit significant DNA binding/cleaving activity, but they served to illustrate the potential for introducing novel amino acid residues within DNA binding protein sequences, and for the development of the tricyclohexyl ester of EDTA as a superior reagent for the introduction of EDT A into synthetic proteins.

Chapter Four describes the discovery and characterization of a new DNA binding/cleaving agent, [SalenMn(III)]OAc. This metal complex produces single- and double-strand cleavage of DNA, with specificity for A:T rich regions, in the presence of oxygen atom donors such as iodosyl benzene, hydrogen peroxide, or peracids. Maximal cleavage by [SalenMn(III)]OAc was produced at pH 6-7. A comparison of DNA singleand double-strand cleavage by [SalenMn(III)]⁺ and other small molecules (Methidiumpropyl-EDTA:Fe, Distamycin-EDTA:Fe, Neocarzinostatin, Bleomycin:Fe) is presented. It was found that DNA cleavage by [SalenMn(III)]⁺ did not require the presence of dioxygen, and that base treatment of DNA subsequent to cleavage by [SalenMn(III)]⁺ afforded greater cleavage and alterations in the cleavage patterns. Analysis of DNA products formed upon DNA cleavage by [SalenMn(III)] indicated that cleavage was due to oxidation of the sugar-phosphate backbone of DNA. Several mechanisms consistent with the observed products and reaction requirements are discussed.

Chapter Five describes progress on some additional studies. In one study, the DNA binding/cleaving specificities of Distamycin-EDTA derivatives bearing pyrrole N-isopropyl substituents were found to be the same as those of derivatives bearing pyrrole N-methyl substituents. In a second study, the design of and synthetic progress towards a series of nucleopeptide activators of transcription are presented. Five synthetic plasmids designed to test for activation of in vitro run-off transcription by DNA triple helix-forming oligonucleotides or nucleopeptides are described.

Chapter Six contains the experimental documentation of the thesis work.

Observação do transporte de sedimentos em suspensão ao longo do Canal Norte do rio Amazonas durante condições de baixa descarga (outubro 2008)

The Amazon river, located in northernBrazil, discharges between 80,000 and 250,000 m3s-1 of water onto the adjacent shelf, creating a plume of brackish water that extends hundreds of kilometers away from the river mouth. This river also carries a large amount of fine sediments to the ocean where fluid mud has been found in the topset and upper foreset layers of the subaqueous delta formed on the mid-shelf. One of the main goals of this dissertation is to describe how turbulence and suspended sediment concentration vary along the Northern Channel of the Amazon river. Water column measurements were carried out in October 2008 at six anchor stations (P1, P3, P5, P6, P8 e P9) located seaward of the river mouth; P1 and P9 were 125 km apart. Each station was occupied during 13 hours during which current speed and direction were continuously sampled with a 600 kHz Teledyne-RDI ADCP; hourly profiles of temperature, salinity, turbidity and depth were also obtained. Water samples were collected for determination of Suspended Particulate Matter (SPM) concentration and calibration of the turbidity sensor. Current speed reached values above 1.5 m s1 in the along-channel direction (NE-SW); a remarkable ebb-flood asymmetry was observed and flows were strongly ebb-dominated. Throughout the water column, SPM concentration at stations P1 and P3 varied between 100 and 300 mg L1 in association with the presence of freshwater. In contrast, a strong salinity gradient was observed between stations P6 and P9, coinciding with the occurrence of concentrations of SPM above 10 g L-1 (fluid mud). At stations P3, P5 and P6, interface between freshwater from the Amazon river and salt water from the continental shelf, shear stresses wereestimated through four diferents methods: Reynolds, Turbulent Kinetic Energy (TKE), modified TKE and Quadratic Law; in the nearbed region (3 mab) the computed values varied between 0 and 3 Pa. At the three stations (P3, P5 and P6) the lowest and the highest shear stress values were obtained through, respectively, the Reynolds and the TKE methods. Over the whole water column turbulence intensity was estimated through the standard deviation of the turbulent component of the along-channel current velocity (root-mean square of u); from these values, it was estimated the turbulent dissipation of energy (G), whose values at 3 mab varied between zero and 20 s1.

Participação do estresse oxidativo na lesão pulmonar induzida por lipopolissacarídeo: repercussões inflamatórias estruturais e funcionais

O objetivo do presente estudo foi investigar o envolvimento do estresse oxidativo na lesão pulmonar aguda (LPA) induzida por lipopolissacarídeo (LPS) e as repercussões inflamatórias, estruturais e funcionais, através de análises bioquímicas de estresse oxidativo, prova de função pulmonar, análise histológica e RT-PCR para citocinas e fatores de transcrição pró-inflamatórios. Na primeira etapa foram utilizados camundongos machos C57BL6 foram divididos em sete grupos: Grupo controle (CTR) (50 μL de solução fisiológica) administrados via intratraqueal [it], LPS 6 horas (10 μL de LPS) [it], LPS 12 horas (10 μL de LPS) [it], LPS 24 horas (10 μL de LPS) [i], LPS 48 horas (10 μL de LPS). Para verificar que as alterações observadas eram estresse oxidativo dependentes camundongos machos C57BL6 foram pré-tratados com N-acetilcisteína (NAC) 1 hora antes do estímulo com LPS e sacrifícados 24 horas depois do estímulo com LPS. Os animais foram divididos da seguinte forma: Grupo LPS 24 horas (10 μL) [it], grupo NAC 40 mg/kg (gavagem) + LPS (10 μL) [it] e grupo NAC 100 mg/kg (gavagem) + LPS (10 μL) [it]. O sistema antioxidante enzimático protegeu o pulmão do estresse oxidativo nas primeiras 12 horas. O estresse oxidativo foi caracterizado em 24 horas e em 48 horas observou-se falência do sistema antioxidante enzimático. Parâmetros de função pulmonar se mostraram alterados nos grupo estimulados com LPS principalmente no grupo LPS. A elastância (p<0,001), resistência de via aérea periférica (ΔP2) (p<0,001), resistência de via aérea central (ΔP1) (p<0,001) e resistência de via aérea total (ΔPtot) (p<0,001) se mostraram principlamente alteradas no grupo LPS 24 horas. O pré-tratamento com NAC impediu o aumento dos parâmetros de elastância (p<0,001), resistência de via aérea periférica (ΔP2) (p<0,001) resistência de via aérea central (ΔP1) (p<0,05) e resistência de via aérea total (ΔPtot) (p<0,001) comparado com o grupo LPS 24 horas. As alterações histológicas como espessamento de septo alveolar, influxo de células inflamatórias e hemorragia mostraram-se tempo dependentes. O pré-tratamento NAC impediu as alterações observadas nos grupo estimulados com LPS. Alterações inflamatórias foram observadas no grupo estimulado com LPS como IL-6 (p<0,001), iNOS (p<0,001), COX2 (p<0,05), TNF-α (p<0,001) e NFκB (p<0,001) quando comparados ao grupo controle. O pré-tratamento com NAC impediu o aumento desses parâmetros como IL-6 (p<0,001), iNOS (p<0,001), COX2 (p<0,05), TNF-α (p<0,05) e NFκB (p<0,001) quando comparados ao grupo LPS 24 horas. Nossos resultados sugerem que o estresse oxidativo desempenha um papel importante nas respostas inflamatórios, estruturais e funcionais no modelo de LPA induzido por LPS e que essas alterações são estresse oxidativo dependentes.

Progressive Decrease in the Potential Usefulness of Meningococcal Serogroup B Vaccine (4CMenB, Bexsero (R)) in Gipuzkoa, Northern Spain

The effectiveness of a vaccine is determined not only by the immunogenicity of its components, but especially by how widely it covers the disease-causing strains circulating in a given region. Because vaccine coverage varies over time, this study aimed to detect possible changes that could affect vaccine protection during a specific period in a southern European region. The 4CMenB vaccine is licensed for use in Europe, Canada, and Australia and is mainly directed against Neisseria meningitidis serogroup B. This vaccine contains four main immunogenic components: three recombinant proteins, FHbp, Nhba and NadA, and an outer membrane vesicle [PorA P1.4]. The allelic distribution of FHbp, Nhba, NadA, and PorA antigens in 82 invasive isolates (B and non-B serogroups) isolated from January 2008 to December 2013 were analyzed. 4CMenB was likely protective against 61.8% and 50% of serogroup B and non-B meningococci, respectively, in the entire period, but between 2012 and 2013, the predicted protection fell below 45% (42.1% for serogroup B isolates). The observed decreasing trend in the predicted protection during the 6 years of the study (X-2 for trend = 4.68, p=0.03) coincided with a progressive decrease of several clonal complexes (e. g., cc11, cc32 and cc41/44), which had one or more antigens against which the vaccine would offer protection.

Aplicação de Inteligência Computacional para a Solução de Problemas Inversos de Transferência Radiativa em Meios Participantes Unidimensionais

Esta pesquisa consiste na solução do problema inverso de transferência radiativa para um meio participante (emissor, absorvedor e/ou espalhador) homogêneo unidimensional em uma camada, usando-se a combinação de rede neural artificial (RNA) com técnicas de otimização. A saída da RNA, devidamente treinada, apresenta os valores das propriedades radiativas [ω, τ0, ρ1 e ρ2] que são otimizadas através das seguintes técnicas: Particle Collision Algorithm (PCA), Algoritmos Genéticos (AG), Greedy Randomized Adaptive Search Procedure (GRASP) e Busca Tabu (BT). Os dados usados no treinamento da RNA são sintéticos, gerados através do problema direto sem a introdução de ruído. Os resultados obtidos unicamente pela RNA, apresentam um erro médio percentual menor que 1,64%, seria satisfatório, todavia para o tratamento usando-se as quatro técnicas de otimização citadas anteriormente, os resultados tornaram-se ainda melhores com erros percentuais menores que 0,04%, especialmente quando a otimização é feita por AG.

Estudo da cromatina nos sítios de fixação à matriz nuclear no domínio do gene TP53 e das modificações epigenéticas e no modelo de progressão tumoral mamária 21T

Dentre os diversos tipos de câncer agressivos, o câncer de mama é o mais comum em mulheres. Mutações hereditárias e adquiridas, assim como alterações epigenéticas atuam em sinergia na carcinogênese mamária e na progressão tumoral. A proteína P53 é uma supressora de tumor e possui uma atuação fundamental na integridade genômica. Apesar do vasto conhecimento sobre o controle da P53 a nível de proteína, ainda pouco se sabe sobre o controle transcricional do gene TP53. A série 21T, uma série de 4 linhagens celulares originadas da mama da mesma paciente, representando diferentes estágios de progressão tumoral mamária, é um eficiente modelo para investigação das alterações epigenéticas e suas influências na expressão gênica ao longo da progressão do câncer de mama. Nós analisamos a organização do domínio do gene TP53 através da técnica de arranjo de DNA, em diversas linhagens celulares de câncer de mama e linhagens controle, e realizamos uma tentativa de caracterizar estes elementos de DNA nas linhagens controle não-tumorais HB2 e MCF10A e nas tumorais MCF-7, MDA-MB-231, T47D, através dos marcadores epigenéticos de eucromatina, H4Ac, e heterocromatina, H3K9me3. Ainda analisamos a ligação de proteínas à região associada à matriz nuclear (MAR), denominada MAR 2, e a possível ligação da proteína ligante à matriz nuclear (MARBP), PARP-1, através de ensaios de gel shift (EMSA). Detectamos que na linhagem controle epitelial mamária, HB2, o gene TP53 está posicionado num domínio de DNA relativamente pequeno, aproximadamente 50 kb, delimitado por dois sítios de fixação à matriz nuclear. Interessantemente, esta estrutura de domínio se apresentou radicalmente diferente nas linhagens de câncer de mama estudadas, MCF7, T47D, MDA-MB-231 e BT474, nos quais o tamanho do domínio estudado estava aumentado e a transcrição do TP53 diminuída. Os enriquecimentos com os marcadores epigenéticos de cromatina H4Ac e H3K9me3 estão diferentemente distribuídos nas MARs nas linhagens celulares. Surpreendentemente, a MAR 2 apresentou uma ligação altamente específica, o que poderia representar a atuação de fatores transcricionais envolvidos na organização da cromatina. Através de programas de bioinformática, detectamos putativos sítios para interessantes fatores de transcrição, tais como o c/EBP-beta e c-myb, que poderiam atuar em cis regulando a expressão do gene TP53 e outros flanqueadores. Nós propusemos um modelo para a organização da cromatina na região de domínio do gene TP53 com os genes flanqueadores. Através da série 21T, detectamos uma hipometilação global genômica, nas células cancerosas 21NT e 21MT1. Uma importante diminuição da expressão global do marcador H4Ac nas células metastáticas 21MT1, foi detectada em relação às outras linhagens. Os níveis de RNAm das principais enzimas relacionadas as modificações epigenéticas são consistentes com as observadas hipometilação genômica e hipoacetilação. Através de microscopia confocal, verificamos que o marcador H4Ac está localizado, na maior parte na periferia e o marcador H3K9me3, pericêntrico nos núcleos tumorais. Por fim, verificamos que o promotor P1 do gene TP53 apresenta um estado de cromatina aberta, e a expressão do gene TP53 é similar em todas as células da série 21T.

滇西北地区晚更新世以来植被演替与气候变化研究

随着全球环境的变化，人们迫切的希望了解整个地球环境的变化过程、规律和未来发展的趋势。对地质历史时期发生过的环境事件的了解，能为我们研究现在的和预测未来的环境变化提供有价值的资料。 孢粉分析作为一种古环境的代用指标，在恢复古植被与古气候方面起着不可替代的作用。而精确的孢粉分析建立在对现代花粉与当地植被的关系正确的认识基础上，由此才可以更准确地提取地层中的花粉所蕴含的植被和环境信息。 云南省地理位置较特殊，地貌复杂，气候类型多样，与之对应的植物种类多样性丰富，植被类型多样。本项研究选择云南省南部和西部地区作为研究地点，采集表土、地表苔藓、蜘蛛网、树皮和树上地衣苔藓五种天然孢粉捕捉器来研究现代花粉与当地植被的关系，为认识现代花粉雨提供新的研究思路。同时，我们在滇西北选择拉市海、文海、哈里谷和属都湖这四个高原湖泊来研究晚更新世以来滇西北地区植被演替和气候变化，为云南历史时期气候变化提供新的研究资料，同时也为全面认识晚更新世以来的气候变化提供新的证据。研究结果如下： 1. 现代花粉雨与当地植被关系的研究 对云南省西北部、中部和南部采集的19个蜘蛛网样品分析得出：云南省西北部样品中共鉴定20个孢粉类型，分属于16个科;云南省中部样品中共鉴定28个类型，分属于23个科;在云南省南部勐腊县采集的样品中共鉴定28个类型，分属于25个科;在望天树北京植物园中采集的样品共鉴定38个类型，分属于34个科;在西双版纳北京植物园中采集的样品中共有11个类型，分属于10个科。结果表明：蜘蛛网样品中分析出的孢粉类型反映了云南省从南到北不同的植被类型，说明蜘蛛网可以作为一种天然孢粉捕捉器来进行现代花粉雨的研究。 同时，在云南西北部文笔水库采集的五种天然孢粉捕捉器（表土、地表苔藓、树皮、蜘蛛网、树上地衣和苔藓）进行了对比研究。结果表明：表土所得的孢粉中木本植物花粉的百分含量为90.5-96%，远大于地表苔藓（53.1-81.7%）、蜘蛛网（61.7%）、树皮（53.8%）和树上地衣、苔藓（50-53.6%）所获得的木本植物花粉的百分含量。与当地植被进行对比结果表明：这五种天然孢粉捕捉器中，表土、地表苔藓和蜘蛛网比树皮和树上地衣、苔藓能更好的反映当地的植被。 2. 滇西北地区晚更新世以来植被演替与气候变化 在过去2, 5000年左右的地质历史时期，滇西北地区植被和气候变化大致经历了以下三个发展阶段。 （1） 2, 5381 B.P—1, 9335 B.P.：属都湖地区（海拔：3620m）主要是以蓼科植物为主的草甸植被，气候寒冷湿润。 （2） 1, 9335 B.P.—1, 2426 B.P.：属都湖地区是以藜科或蒿属为主的草甸植被，气候从湿润过渡到干旱;而在海拔稍低的文海地区（海拔3080m），主要是阔叶栎林或针阔混交林，气候温凉略湿到寒冷干旱。 （3） 1, 2426 B.P.—至今：属都湖地区是以蓼科为主的草甸植被，气候仍寒冷湿润;海拔稍低的哈里谷（海拔3277m）是以松和冷杉为主的针叶林，气候冷干;文海地区的植被逐渐演替为以松和冷杉为主的针阔叶混交林，但栎属花粉的含量远高于哈里谷地区，说明当时文海的气温虽然低，却比哈里谷稍高;而海拔更低的拉市海（海拔2440m）则是以松为主的针叶林，气候比其他几个地区温暖。 在全新世气候最适宜期（约8000 B.P.以来），哈里谷是以栎和松为主的针阔混交林;文海是以松为主的针阔混交林;而拉市海是以松和蕨类植物为主。说明随着海拔的升高（从拉市海到属都湖），温度逐渐下降，植被类型也由暖性针叶林过渡到针阔混交林再到草甸。 从6000 B.P.至今，文海和哈里谷植被在这一阶段都是以松和冷杉为主的暗针叶林，少量混生一些阔叶类植物，气候变得冷湿;属都湖则仍是以蓼科为主的草甸，气候变得更加寒冷。 研究证明：滇西北地区总的气候变化趋势与全球变化规律基本一致，在1,1805—9990 B.P.期间发生新仙女木事件，在8000B.P.以来，气候变得温暖湿润，为全新世气候最适宜期。此后，气候逐渐变得接近现代。

Trends in counts of manatees Trichechus manatus latirostris from 1987 to 2006 in waters of Sarasota County, Florida, USA

To demonstrate the utility of distributional surveys for assessing relative abundance and trends in counts for a discrete area of coastline, aerial survey data from Sarasota County, Florida, USA, were analyzed for the years 1987 to 2006. The study area was divided into 3 regions: the Sarasota Bay Region (SBR; N = 353 surveys), Lemon Bay (N = 368), and the Myakka River (N = 209). Manatee counts varied significantly across seasons (p < 0.0001) for all 3 regions. Manatees within Sarasota County utilized open bays primarily in the warmer months. Such usage may have been influenced by resource availability. Conversely, usage of the Myakka River peaked in winter months when manatees seek warm-water refugia such as Warm Mineral Spring. Marginal means for yearly counts within Lemon Bay and the SBR increased significantly, beginning midway through the survey period (1996) until the early 2000s. In contrast, mean yearly counts within the Myakka River decreased over this time period. After record lows in 2003 for Lemon Bay and the Myakka River, and a considerable decline in 2004 for the SBR, mean yearly counts for all 3 regions showed an increasing trend over the remaining 2 yr of the study. Greater protection of manatee habitat and availability of forage coincided with the increase in numbers of manatees using Sarasota County waters during the 1990s, and the subsequent decline in numbers may be indicative of the increase in mortality in recent years due to watercraft collisions and severe red tide events.