Generation and characterization of ionic and neutral dihydroxy boron B(OH)(2)(+/0) in the gas phase

The dicoordinated borinium ion, dihydroxyborinium, B(OH)(2)(+) is generated from methyl boronic acid CH3B(OH)(2) by dissociative electron ionization and its connectivity confirmed by collisional activation. Neutralization-reionization (NR) experiments on this ion indicate that the neutral B(OH)(2) radical is a viable species in the gas phase. Both vertical neutralization of B(OH)(2)(+) and reionization of B(OH)(2) in the NR experiment are, however, associated with particularly unfavorable Franck-Condon factors. The differences in adiabatic and vertical electron transfer behavior can be traced back to a particular pi stabilization of the cationic species compared to the sp(2)-type neutral radical. Thermochemical data on several neutral and cationic boron compounds are presented based on calculations performed at the G2 level of theory.

The central region of meso-3,4-diphenylhexane-2,5-dione

As with 1,2-diphenylethane (dpe), X-ray crystallographic methods measure the central bond in meso-3,4-diphenylhexane-2,5-done (dphd) as significantly shorter than normal for an sp(3)-sp(3) bond. The same methods measure the benzylic (ethane C-Ph) bonds in dphd as unusually long for sp(3)-sp(2) liaisons. Torsional motions of the phenyl rings about the C-Ph bonds have been proposed as the artifacts behind the result of a 'short' central bond in dpe. While a similar explanation can, presumably, hold for the even 'shorter' central bond in dphd, it cannot account for the 'long' C-Ph bonds. The phenyl groups, departing much from regular hexagonal shape, adopt highly skewed conformations with respect to the plane constituted by the four central atoms. It is thought that-the thermal motions of the phenyl rings, conditioned by the potential wells in which they are ensconced in the unit cell, are largely libratory around their normal axes. In what appears to be a straightforward explanation under the 'rigid-body' concept, it appears that these libratory motions of the phenyl rings, that account, at the same time, for the 'short' central bond, are the artifacts behind the 'long' measurement of the C-Ph bonds. These motions could be superimposed on torsional motions analogous to those proposed in the case of dpe. An inspection of the ORTEP diagram from the 298 K data on dphd clearly suggests these possibilities. Supportive evidence for these qualitative explanations from an analysis of the differences between the mean square displacements of C(1) and C(7)/C(1a) and C(7a) based on the 'rigid-body model' is discussed. (C) 2002 Elsevier Science B.V. All rights reserved.

Selective Orthosteric Free Fatty Acid Receptor 2 (FFA2) Agonists IDENTIFICATION OF THE STRUCTURAL AND CHEMICAL REQUIREMENTS FOR SELECTIVE ACTIVATION OF FFA2 VERSUS FFA3

Free fatty acid receptor 2 (FFA2; GPR43) is a G protein-coupled seven-transmembrane receptor for short-chain fatty acids (SCFAs) that is implicated in inflammatory and metabolic disorders. The SCFA propionate has close to optimal ligand efficiency for FFA2 and can hence be considered as highly potent given its size. Propionate, however, does not discriminate between FFA2 and the closely related receptor FFA3 (GPR41). To identify FFA2-selective ligands and understand the molecular basis for FFA2 selectivity, a targeted library of small carboxylic acids was examined using holistic, label-free dynamic mass redistribution technology for primary screening and the receptor-proximal G protein [S-35] guanosine 5'-(3-O-thio) triphosphate activation, inositol phosphate, and cAMP accumulation assays for hit confirmation. Structure-activity relationship analysis allowed formulation of a general rule to predict selectivity for small carboxylic acids at the orthosteric binding site where ligands with substituted sp(3)-hybridized alpha-carbons preferentially activate FFA3, whereas ligands with sp(2)- or sp-hybridized alpha-carbons prefer FFA2. The orthosteric binding mode was verified by site-directed mutagenesis: replacement of orthosteric site arginine residues by alanine in FFA2 prevented ligand binding, and molecular modeling predicted the detailed mode of binding. Based on this, selective mutation of three residues to their non-conserved counterparts in FFA3 was sufficient to transfer FFA3 selectivity to FFA2. Thus, selective activation of FFA2 via the orthosteric site is achievable with rather small ligands, a finding with significant implications for the rational design of therapeutic compounds selectively targeting the SCFA receptors.

Interactions between the powdery mildew effector BEC1054 and barley proteins identify candidate host targets

There are over 500 candidate secreted effector proteins (CSEPs) or Blumeria effector candidates (BECs) specific to the barley powdery mildew pathogen Blumeria graminis f.sp. hordei. The CSEP/BEC proteins are expressed and predicted to be secreted by biotrophic feeding structures called haustoria. Eight BECs are required for the formation of functional haustoria. These include the RNase-like effector BEC1054 (synonym CSEP0064). In order to identify host proteins targeted by BEC1054, recombinant BEC1054 was expressed in E. coli, solubilized, and used in pull-down assays from barley protein extracts. Many putative interactors were identified by LC-MS/MS after subtraction of unspecific binders in negative controls. Therefore, a directed yeast-2-hybrid assay, developed to measure the effectiveness of the interactions in yeast, was used to validate putative interactors. We conclude that BEC1054 may target several host proteins, including a glutathione-S-transferase, a malate dehydrogenase, and a pathogen-related-5 protein isoform, indicating a possible role for BEC1054 in compromising well-known key players of defense and response to pathogens. In addition, BEC1054 interacts with an elongation factor 1 gamma. This study already suggests that BEC1054 plays a central role in barley powdery mildew virulence by acting at several levels.

Concentração de potássio na solução nutritiva e a qualidade e número de frutos de melão por planta em hidroponia

O experimento foi conduzido em hidroponia (NFT) no período de junho a novembro de 2001, na FCAV-UNESP em Jaboticabal-SP, situada a 21º15 22 Sul, 48º18 58 Oeste, e altitude de 575 metros. O híbrido Bônus n0 2 foi cultivado em blocos casualizados, em esquema de parcelas subdivididas, com seis repetições. Os elementos avaliados foram número de frutos por planta (2, 3, 4 e fixação livre) e concentração de potássio na solução nutritiva (66; 115,5; 165 e 247,5mg L-1). O experimento teve como objetivo avaliar a concentração de potássio na solução nutritiva e número de frutos por planta sobre a qualidade dos frutos do meloeiro. Concentrações maiores do que 66mg L-1 de potássio na solução nutritiva não promoveram incrementos nas características de qualidade dos frutos do meloeiro como: teor de sólidos solúveis médio de todos os frutos da planta, do segundo fruto, da acidez total titulável média de todos os frutos da planta, do primeiro fruto, do segundo fruto, do índice de maturação, da espessura da polpa e do índice de formato dos frutos. Frutos oriundos de plantas de melão com dois frutos apresentaram maior teor de sólidos solúveis e acidez total titulável, independente da concentração de potássio na solução nutritiva. O maior número de frutos por planta reduziu a espessura do mesocarpo dos frutos.

Efeito da concentração de nitrogênio na solução nutritiva e do número de frutos por planta sobre a produção do meloeiro

O trabalho foi conduzido em casa de vegetação, na UNESP em Jaboticabal (SP), de junho a novembro de 2001, com o objetivo de avaliar a produção do melão (Cucumis melo var. reticulatus), híbrido Bônus nº2, cultivado em sistema hidropônico NFT, em função da concentração de nitrogênio na solução nutritiva (80, 140, 200 e 300 mg L-1) e número de frutos por planta (2, 3, 4 e livre). O delineamento experimental utilizado foi o de blocos ao acaso, em parcelas subdivididas, com seis repetições. Aos 80 dias após o transplantio, foram observados 2, 3, 4 e 5,1 frutos por planta e, posteriormente na colheita, 2, 2,9, 3,0 e 3,4 frutos por planta, respectivamente para os tratamentos com 2, 3, 4 e fixação livre, sendo esta redução atribuída ao abortamento de frutos. Houve redução no peso médio do 1º, 2º e 3º fruto colhido, com o aumento da concentração de nitrogênio. Plantas com o menor número de frutos, apresentaram maior peso médio dos mesmos, porém com menor produção por planta. A maior produção (2.474 g/planta) foi obtida com 80 mg L-1 de nitrogênio na solução nutritiva.

Astyanax baird & girard 1854 (Ostariophysi: Characiformes: Characidae) do sistema do Alto rio Paraná: taxonomia

Pós-graduação em Biologia Animal - IBILCE

Descrição cariotípica de peixes dos gêneros Baryancistrus, Parancistrus, Peckoltia e Ancistrus (Ancistrinae, Loricariidae) da Bacia Amazônica

A subfamília Ancistrinae é uma das mais diversificadas entre os Loricariidae, incluindo cerca de 200 espécies distribuídas em 26 gêneros. Esses peixes são facilmente reconhecidos pela presença de placas ósseas dispostas em séries ao longo do corpo e pela presença de boca em posição ventral anterior. São vulgarmente conhecidos por acaris, bodós, cascudos. As espécies da subfamília Ancistrinae representam um importante recurso sócio-econômico, constituindo uma das mais importantes atividades comerciais no município de Altamira-PA. Foram analisadas, através das técnicas convencionais (Giemsa, bandeamento C e Ag-NORs) e técnica de fluorocromo (Cromomicina A3), dez espécies de peixes da subfamília Ancistrinae pertencentes a quatro gêneros (Baryancistrus, Parancistrus, Peckoltia e Ancistrus). As espécies do gênero Baryancistrus revelaram um número diplóide 2n= 52 e NF=104. A NOR foi encontrada em posição intersticial no braço curto de um par cromossômico do tipo meta/submetacêntrico. A espécie B. aff. niveatus apresentou grandes blocos heterocromáticos ricos em pares de bases G-C como apomorfia, sendo esta espécie considerada como mais derivada cariotipicamente entre os Baryancistrus. As espécies do gênero Parancistrus apresentaram uma estrutura cariotípica muito similar àquela encontrada em Baryancistrus, apresentando as Regiões Organizadoras de Nucléolos como uma provável sinapomorfia entre os dois gêneros. Os representantes do gênero Peckoltia possuem número diplóide 2n=52 e NF=102. Todas as espécies analisadas apresentaram grandes blocos heterocromáticos, envolvendo quase todos os braços longos de alguns pares cromossomos do tipo submetacêntricos e subtelocêntricos, sendo esta característica uma provável sinapornorfia para este grupo. A NOR foi localizada no braço longo de um par de cromossomos submetacêntricos em P. vittata e em no máximo três cromossomos nas espécies Peckoltia sp.1 e Peckoltia sp.2. A espécie Ancistrus ranunculus foi a que apresentou o cariótipo mais derivado entre as espécies estudadas, com o número dipló ide igual a 48 cromossomos e NF 80. As análises citogenéticas feitas até agora sugerem que os principais eventos de diversificação cariotípica para os Ancistrinae foram às inversões, a exceção de Ancistrus ranunculus que apresentou também rearranjos Robertsonianos.

A simple geometrical path towards hybrid orbitals

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Aplicação de reguladores vegetais na produção e qualidade de frutos do tomateiro

Pós-graduação em Agronomia (Horticultura) - FCA

The role of suppressor factors in the regulation of immune responses by ultraviolet radiation -induced suppressor T lymphocytes

The purpose of these studies was to determine the role of suppressor factors (TsF) in the regulation of immune responses by ultraviolet radiation-induced suppressor T lymphocytes (Ts). The Ts were induced following epicutaneous sensitization with contact allergens to an unirradiated site on mice irradiated five days earlier with 40 kJ/m$\sp2$ UVB (280-320 nm) radiation. The spleens of such mice contain afferent, hapten-specific, Thy-1$\sp+$, Lyt-1$\sp+$,2$\sp-$ Ts that suppress in vivo contact hypersensitivity (CHS) and antibody responses and the in vitro generation of cytotoxic T lymphocytes (CTL). Four approaches were used to determine the role of TsF. First, lysates produced from sonically-disrupted Ts were injected i.v. into normal animals; they inhibited CHS in vivo in a nonspecific manner. The lysates suppressed the induction and elicitation of CHS, and they inhibited the in vitro generation of CTL. Lysates prepared from splenocytes obtained from unirradiated mice or UV-irradiated, unsensitized mice failed to inhibit either response. Second, supernatants from cultures containing Ts, normal syngeneic responder lymphocytes, and hapten-modified stimulator cells were injected i.v. into normal recipients. They inhibited the induction of CHS and did so in a hapten-specific manner. Cellular and kinetic requirements were observed for the generation of suppressive activity. Splenocytes from mice treated with Ts supernatants suppressed CHS when transferred into normal animals. The supernatants also suppressed the in vitro generation of specific CTL. Third, the TsF-specific B16G monoclonal antibody was tested for its ability to modulate the effects of UV radiation in vivo. The i.v. injection of B16G into UV-irradiated mice reduced the suppression of CHS. Splenocytes of B16G-treated mice transferred into normal recipients, and they suppressed CHS, indicating that the Ts were not depleted. Fourth, B16G was used to isolate a putative TsF by antibody immunoadsorbance. When the B16G-bound fraction was eluted and injected i.v. into normal animals, it suppressed CHS and represented a 900-fold enrichment of activity over the starting material, based on specific activity. By SDS-PAGE, the B16G-bound material contained nondisulfide-linked 45- and 50-kDa components. These results suggest that TsF may play an immunoregulatory role in CHS. The isolation of a UV radiation-induced TsF lends credence to the involvement of such molecules. ^

Structural determinants of subunit -subunit interactions and subcellular localization of the calcium /calmodulin -dependent protein kinase II

The multifunctional Ca$\sp{2+}$/calmodulin-dependent protein kinase II (CaM kinase) is a Ser/Thr directed protein kinase that participates in diverse Ca$\sp{2+}$ signaling pathways in neurons. The function of CaM kinase depends upon the ability of subunits to form oligomers and to interact with other proteins. Oligomerization is required for autophosphorylation which produces significant functional changes that include Ca$\sp{2+}$/calmodulin-independent activity and calmodulin trapping. Associations with other proteins localize CaM kinase to specific substrates and effectors which serves to optimize the efficiency and speed of signal transduction. In this thesis, we investigate the interactions that underlie the appropriate positioning of CaM kinase activity in cells. We demonstrate that the subcellular distribution of CaM kinase is dynamic in hippocampal slices exposed to anoxic/aglycemic insults and to high K$\sp{+}$-induced depolarization. We determine the localization of CaM kinase domains expressed in neurons and PC-12 cells and find that the C-terminal domain of the $\alpha$ subunit is necessary for localization to dendrites. Moreover, monomeric forms of the enzyme gain access to the nucleus. Attempts made to identify novel CaM kinase binding proteins using the yeast two-hybrid system resulted in the isolation of hundreds of positive clones. Those that have been sequenced are identical to CaM kinase isoforms. Finally, we report the discovery of specific regions within the C-terminal domain that are necessary and sufficient for subunit-subunit interactions. Differences between the $\alpha$ and $\beta$ isoforms were discovered that indicate unique structural requirements for oligomerization. A model for how CaM kinase subunits interact to form holoenzymes and how structural heterogeneity might influence CaM kinase function is presented. ^

Benthic foraminifera abundance data of sediment core GeoB7926-2

We present a high resolution, multiproxy study of the relationship between pelagic and benthic environments of a coastal upwelling system in the subtropical NE Atlantic Ocean. Marine sediments corresponding to late MIS3 to the Holocene in the radiocarbon dated core GeoB7926, retrieved off Mauritania (21°N) were analysed to reconstruct productivity in surface waters and its linkage to deep waters during the last 35 ka BP. High latitude cold events and changes in atmospheric and oceanographic dynamics influenced upwelling intensity over this time period. Subsequently, this caused changes in primary productivity off this low-latitude coastal upwelling locality. The benthic foraminiferal fauna displays four main community shifts corresponding to fundamental climatic events, first of all during late MIS3 (35-28 ka BP), secondly from 28 to 19 ka BP (including Heinrich event 2 and the LGM), thirdly within Heinrich event 1, the Bølling Allerød and the Younger Dryas (18-11.5 ka BP) and finally during the Holocene (11.5-0 ka BP). In particular, strong pelagic-benthic coupling is apparent in MIS 3, as demonstrated by increased primary productivity, indicated by moderate DAR and the dominance of benthic foraminiferal species which prefer fresh phytodetritus. A decline in upwelling intensity and nutrient availability follows, which resulted in a proportionately larger amount of older, degraded matter, provoking a shift in the benthic foraminifera fauna composition. This rapid response of the benthic environment continues with a progressive increase in upwelling intensity due to sea level and oceanographic changes and according high surface production during the LGM. During Heinrich event 1 and the Younger Dryas, extreme levels of primary production actually hindered benthic environment through the development of low oxygen conditions. After this period, a final change in benthic foraminiferal community composition occurs which indicates a return to more oxygenated conditions during the Holocene.

Total and species abundance of Polychaeta during POLARSTERN cruise ANT-XXIV/2, ANDEEP-SYSTCO, in the Southern Ocean

In the course of the ANDEEP-SYSTCO project, during the ANT XXIV-2 expedition in austral summer 2007/2008, the diversity and composition of the Polychaeta of the Antarctic deep-sea and adjacent South Atlantic basins were analyzed. A total of 847 individuals of 31 families were found belonging to 86 different species. Calculation of diversity (Shannon-Wiener Index, Pielou's Evenness) and the general species composition of Polychaeta showed patterns typical for the deep sea, with high species richness and low abundances. Lowest diversity was found in the Agulhas Basin in over 4000 m water depth. Lowest Evenness was found on top of Maud Rise where one-third of all Polychaeta belonged to one species. Cluster analyses resulted in higher affinities of Maud Rise to the Agulhas Basin than to the Antarctic continental slope. Explanations are sought in similarities of environmental factors (e.g., sediment, food input).

Benthic foraminiferal record from Santa Barbara basin

The oxygen minimum zone (OMZ) of the late Quaternary California margin experienced abrupt and dramatic changes in strength and depth in response to changes in intermediate water ventilation, ocean productivity, and climate at orbital through millennial time scales. Expansion and contraction of the OMZ is exhibited at high temporal resolution (107-126 year) by quantitative benthic foraminiferal assemblage changes in two piston cores forming a vertical profile in Santa Barbara Basin (569 m, basin floor; 481 m, near sill depth) to 34 and 24 ka, respectively. Variation in the OMZ is quantified by new benthic foraminiferal groupings and new dissolved oxygen index based on documented relations between species and water-mass oxygen concentrations. Foraminiferal-based paleoenvironmental assessments are integrated with principal component analysis, bioturbation, grain size, CaCO3, total organic carbon, and d13C to reconstruct basin oxygenation history. Fauna responded similarly between the two sites, although with somewhat different magnitude and taxonomic expression. During cool episodes (Younger Dryas and stadials), the water column was well oxygenated, most strongly near the end of the glacial episode (17-16 ka; Heinrich 1). In contrast, the OMZ was strong during warm episodes (Bølling/Allerød, interstadials, and Pre-Boreal). During the Bølling/Allerød, the OMZ shoaled to <360 m of contemporaneous sea level, its greatest vertical expansion of the last glacial cycle. Assemblages were then dominated by Bolivina tumida, reflecting high concentrations of dissolved methane in bottom waters. Short decadal intervals were so severely oxygen-depleted that no benthic foraminifera were present. The middle to late Holocene (6-0 ka) was less dysoxic than the early Holocene.