Structure, genomic DNA typing, and kinetic characterization of the D allozyme of placental alkaline phosphatase (PLAP/ALPP)

The D allozyme of placental alkaline phosphatase (PLAP) displays enzymatic properties at variance with those of the common PLAP allozymes. We have deduced the amino acid sequence of the PLAP D allele by PCR cloning of its gene, ALPP Two coding substitutions were found in comparison With the cDNA of the common PLAP F allele, i.e., 692C>G and 1352A>G, which translate into a P209R and E429G substitution. A single nucleotide primer extension (SNuPE) assay was developed using PCR primers that enable the amplification of a 1.9 kb PLAP fragment. Extension primers were then used on this PCR fragment to detect the 692C>G and 1352A>G substitution. The SNuPE assay on these two nucleotide substitutions enabled us to distinguish the PLAP F and D alleles from the PLAP S/I alleles. Functional studies on the D allozyme were made possible by constructing and expressing a PLAP D cDNA, i.e., [Arg209, Gly429] PLAP, into wildtype Chinese hamster ovary cells. We determined the k(cat) and K-m, of the PLAP S, F. and D allozymes using the non,physiological substrate p-nitrophenylphosphate at an optimal pH (9.8) as well as two physiological substrates, i.e., pyridoxal-5'-phosphate and inorganic pyrophosphate at physiological pH (7.5). We found that the biochemical properties of the D allozyme of PLAP are significantly different from those of the common PLAP allozymes. These biochemical findings suggest that a suboptimal enzymatic function by the PLAP D allozyme may be the basis for the apparent negative selective pressure of the PLAP D allele. The development of the SNuPE assay will enable us to test the hypothesis that the PLAP D allele is subjected to intrauterine selection by examining genomic DNA from statistically informative population samples. Hum Mutat 19:258-267, 2002. (C) 2002 Wiley-Liss, Inc.

Quantization of the superstring with manifest U(5) super-Poincaré invariance

The superstring is quantized in a manner which manifestly preserves a U(5) subgroup of the (Wick-rotated) ten-dimensional super-Poincaré invariance. This description of the superstring contains critical N = 2 worldsheet superconformal invariance and is a natural covariantization of the U(4)-invariant light-cone Green-Schwarz description. © 1999 Published by Elsevier Science B.V. All rights reserved.

Proposal of a new technique for bile duct reconstruction after iatrogenic injury. Study in dogs and review of the literature

Purpose: Interposition of a jejunal tube between the common bile duct and duodenum. Methods: Five adult mongrel dogs of both sexes, weighing on average 22.3 kg (18 to 26.5 kg), were used. Obstructive jaundice was induced by ligation of the distal common bile duct. After one week, a 2.5-cm long jejunal tube was fabricated from a segment of the loop removed 15 cm from the Treitz angle and interposed between the common bile duct and duodenum. Results: The animals presented good clinical evolution and no complications were observed. After 6 weeks, complete integration was noted between the bile duct mucosa, tube and duodenum and a significant reduction in total bilirubin and alkaline phosphatase was observed when compared to the values obtained one week after ligation of the common bile duct. Conclusion: The jejunal tube interposed between the dilated bile duct and duodenum showed good anatomic integration and reduced total bilirubin and alkaline phosphatase levels in the animals studied.

On the influence of the mode-shapes of a marine propulsion shafting system on the prediction of torsional stresses

Torsional vibration predictions and measurements of a marine propulsion system, which has both damping and a highly flexible coupling, are presented in this paper. Using the conventional approach to stress prediction in the shafting system, the numerical predictions and the experimental torsional vibration stress curves in some parts of the shafting system are found to be quite different. The free torsional vibration characteristics and forced torsional vibration response of the system are analyzed in detail to investigate this phenomenon. It is found that the second to fourth natural modes of the shafting system have significant local deformation. This results in large torsional resonant responses in different sections of the system corresponding to different engine speeds. The results show that when there is significant local deformation in the shafting system for different modes, then multi-point measurements should be made, rather than the conventional method of using a single measurement at the free end of the shaft, to obtain the full torsional vibration characteristics of the shafting system.

A new species of the oligotrophic genus ochroconis (Sympoventuriaceae)

Ochroconis globalis, a novel species of the melanized genus Ochroconis (Sympoventuriaceae, Venturiales), is described and illustrated and distinguished phenotypically and molecularly from existing ochroconis-like species. Phylogenetic analyses using nuclear ribosomal DNA genes (nuSSU, ITS, nuLSU) and coding gene fragments (ACT1, BT2, TEF1) revealed clustering of all strains as a monophyletic clade which was well separated from known Ochroconis species. Most strains of the new species were obtained from human-made environments, while the natural habitat of the species remains enigmatic. The new combination Ochroconis musae is introduced for one of the most commonly encountered Ochroconis species, and a phenotypic key to all species is provided.

Triatominae survey (Hemiptera: Reduviidae: Triatominae) in the south-central region of the state of Bahia, Brazil between 2008 and 2013

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Soil physical and microbiological attributes cultivated with the common bean under two management systems

Agricultural management systems can alter the physical and biological soil quality, interfering with crop development. The objective of this study was to evaluate the physical and microbiological attributes of a Red Latosol, and its relationship to the biometric parameters of the common bean (Phaseolus vulgaris), irrigated and grown under two management systems (conventional tillage and direct seeding), in Campinas in the state of Sao Paulo, Brazil. The experimental design was of randomised blocks, with a split-plot arrangement for the management system and soil depth, analysed during the 2006/7 and 2007/8 harvest seasons, with 4 replications. The soil physical and microbiological attributes were evaluated at depths of 0.00-0.05, 0.05-0.10, 0.10-0.20 and 0.20-0.40 m. The following were determined for the crop: density, number of pods per plant, number of beans per pod, thousand seed weight, total weight of the shoots and harvest index. Direct seeding resulted in a lower soil physical quality at a depth of 0.00-0.05 m compared to conventional tillage, while the opposite occurred at a depth of 0.05-0.10 m. The direct seeding showed higher soil biological quality, mainly indicated by the microbial biomass nitrogen, basal respiration and metabolic quotient. The biometric parameters in the bean were higher under the direct seeding compared to conventional tillage.

Reproductive indices in natural nests of giant Amazon river turtles Podocnemis expansa (Schweigger, 1812) (Testudines, Podocnemididae) in the Environmental Protection Area Meanders of the Araguaia river

Realizou-se a contagem dos ovos não eclodidos, dos filhotes vivos e mortos de Podocnemis expansa oriundos de 327 ninhos naturais, localizados nas praias da Área de Proteção Ambiental (APA) - Meandros do Rio Araguaia, onde se determinou a porcentagem de eclosão dos ovos (94,63%); não eclosão (5,37%); sobrevivência (94,24%) e mortalidade dos filhotes (5,76%), e a média de filhotes mortos durante os 15 dias no berçário (0,97%). A média do total de filhotes por ninho foi determinada pela soma do número de filhotes vivos e mortos divididos pelo total de ninhos, enquanto que a média do total de ovos por ninho foi determinada pela soma do número de filhotes vivos, mortos e ovos não eclodidos divididos pelo total de ninhos. Com isso, obtiveram-se os valores médios do número de filhotes vivos (88,98 ± 23,94); mortos (0,37 ± 0,93); ovos não eclodidos (5,07 ± 9,57), e total de ovos (94,42 ± 21,30). A eficiência reprodutiva da população selvagem de P. expansa pode ser afetada por muitos fatores ambientais, como temperatura, umidade e precipitação. Além disso, fatores influenciados pelo homem, como a presença de produtos químicos na água e a possibilidade de doenças infecciosas, também têm impacto significativo. Os dados dos índices reprodutivos obtidos neste estudo são indispensáveis para futuras investigações de anomalias de incubação.

The interaction between heparin and Lys49 phospholipase A(2) reveals the natural binding of heparin on the enzyme

We have studied at a molecular level the interaction of heparins on bothropstoxin-1 (BthTx-1), a phospholipase A(2) toxin. The protein was monitored using gel filtration chromatography, dynamic light scattering (DLS), circular dichroism (CD), attenuated total reflectance Fourier transform infrared (ATR-FTIR) and intrinsic tryptophan fluorescence emission (ITFE) spectroscopy. The elution profile of the protein presents a displacement of the protein peak to larger complexes when interacting with higher concentration of heparin. The DLS results shows two R-h at a molar ratio of 1, one to the distribution of the protein and the second for the action of heparin on BthTx-I structures, and a large distribution with the increase of protein. The interaction is accompanied by significant changes in the CD spectra, showing two common features: a decrease in signal at 208 nm (3 and 6 kDa heparins) and an isodichroic point near 226 nm (3 kDa heparin). FTIR spectra indicate that only a few amino acid residues are involved in this interaction. Alterations in the ITFE by binding heparins suggest that the initial binding occurs on the ventral face of BthTx-1. Together, these results add an experimental and structural basis on the action mechanism of the heparins over the phospholipases A(2) and provide a molecular model to elucidate the interaction of the enzyme-heparin complex at a molecular level. (c) 2005 Elsevier B.V. All rights reserved.

Study of the thermomechanical and electrical properties of conducting composites containing natural rubber and carbon black

This work describes the preparation and characterization of composite materials obtained by the combination of natural rubber (NR) and carbon black (CB) in different percentages, aiming to improve their mechanical properties, processability, and electrical conductivity, aiming future applications as transducer in pressure sensors. The composites NR/CB were characterized through optical microscopy (OM), DC conductivity, thermal analysis using differential scanning calorimetry (DSC), dynamic mechanical thermal analysis (DMA), thermogravimetry (TGA), and stress-strain test. The electrical conductivity varied between 10(-9) and 10 S m(-1), depending on the percentage of CB in the composite. Furthermore, a linear (and reversible) dependence of the conductivity on the applied pressure between 0 and 1.6 MPa was observed for the sample with containing 80 wt % of NR and 20% of CB. (C) 2007 Wiley Periodicals, Inc.

Measurement of the natural variation of 13C:12C ratio in leaves at Reserva Ducke Forest, central Amazonia

Investigation of carbon isotope fractionation by plants was carried out at two sub-areas located in Reserva Ducke, central Amazonia: open reserve (virgin forest with low density of plant species); and closed reserve (virgin forest with high density of plant species). Preliminary results (δ‰ 13C: 12C values, PDB) of leaf analysis at different plant heights indicate the following: Eschweilera matamata Hub. (Lecythidaceae), common name 'matamata', -31.55±0.61; Protium heptaplyllum March. (Burseraceae), common name 'breu branco', -32.34±1.39; Calophyllum brasiliense Camb. (Guttiferae), common name 'jacareúba', -30.72±0.23; Scleronema micrantthum Ducke. (Bombacaceae), common name 'cardeiro'. -28.81±0.68; and Carapa guianensis Aubl. (Meliaceae), common name 'andiroba', -31.07±0.51. It is possible that the plant species analysed belong to the C3 photosynthetic cycle. In general, the species in the open reserve show differences of the order of 1.66±0.34‰ (greater in 13C) as compared with the same species in the closed reserve. The old leaves show differences in the relative isotopic enrichment (δ) of the order of 1‰, being smaller in new leaves in both reserves. The probable occurrence of an isotopic gradient from the lower (2-5 m) to the upper part (15-20 m) of the plant, of the order of 1.3‰, smaller in 13C, in species from the dense forest was noted. However, only two plants from each species were analysed during a two-year period, data obtained to far are still preliminary, and results should, therefore, be revised. Moreover, according to the literature, the natural carbon isotope fractionation by plants shows metabolic, physiological and environmental dependence. © 1991.