Baupain, a plant cysteine proteinase that hinders thrombin-induced human platelet aggregation

Bauninia forficata is trivially known as cow paw, and popularly used in Brazil for treatment of diabetes mellitus. Denominated baupain a cysteine proteinase was purified from B. forficata leaves. In this study, we investigated the baupain effect on aggregation of isolated human platelets in vitro and the results show that baupain hinders thrombin - but not ADP- and collagen- induced platelet aggregation. With synthetic quenched-fluorescent peptides, the kinetics of the cleavage site of human proteinase-activated receptor 1 / 2 / 3 and 4 [PAR-1 / 2 / 3 and 4] by baupain was determined. In conclusion, similar to bromelain and papain, baupain hinders human platelets aggregation, probably through an unspecific cleavage in the Phe-Leu bond of PAR1.     

Bioavailability and dosing strategies of mineral in anaerobic mono‐digestion of maize straw

The influence of the bonding form distribution of Fe, Ni, Co and Mn and their potential bioavailability during the anaerobic degradation of maize straw was investigated. Two reactors were operated over 117 days at 37°C and different dosage strategies of mineral were studied in reactor (R2). Control reactor (R1) was metal‐limited over time. mineral supplementation (1 g L^?1) once a week reported the highest methane yield (257 mL g^?1 VS) with 30% of increment. Ni and Co predominated in their oxidizable bonding forms and Fe mainly existed as residual and oxidizable fractions. The potential bioavailability (Mn ?? Co ≈ Ni ? Fe) of R2 was higher comparing to R1. Metal deprivation in R1 led to depletion of both sequential extraction fractions and total metal concentrations until the end of the process. This study confirmed that the dosage strategy of mineral has a stimulatory effect on methane production from crop maize waste.     

Dieback of riparian alder caused by the Phytophthora alni complex: projected consequences for stream ecosystems

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Spherical vs. granular immobilization support selection and performance on an optical flow cell immunosensor based on the fluorescence of Cyanine-5

A spherical porous glass support Trisoperl (TRISO) with four pore diameters (? 47.8; 55.9; 102.6, and 108.8 nm) was characterized and selected for application in an optical flow cell immunosensor, in comparison with controlled pore glass (CPG). The TRISO support was functionalized with aldehyde and isothiocyanate (-NCS) groups to attach bovine serum albumin and alkaline phosphatase (AP). The TRISO isothiocyanate pore diameter 47.8 nm (TRISO(-NCS) 47.8 nm) showed the better potential to be used in the immunosensor. It immobilized more protein (19.3 mg AP per g support) while presenting an optical performance comparable to the CPG. CPG(-NCS) and TRISO(-NCS) 47.8 nm were tested in the immunosensor model where the saturation of the Goat IgG immobilized in the supports with Monoclonal Anti-Goat IgG conjugated with Cyanine-5 was reached, followed by regeneration with the elution buffer modified PBS pH 2.0. The TRISO(-NCS) 47.8 nm presented lower fluorescence intensity at saturation (around 39 AU) than CPG(-NCS) (150 to 104 AU), but revealed a major advantage related to the uniform arrangement of the spherical particles in the flow cell, generating no significant fluorescence differences between gravity and flow package.     

Cnr interferes with dimerization of the replication protein alpha in phage-plasmid P4

DNA replication of phage-plasmid P4 in its host Escherichia coli depends on its replication protein α. In the plasmid state, P4 copy number is controlled by the regulator protein Cnr (copy number regulation). Mutations in α (α^cr) that prevent regulation by Cnr cause P4 over-replication and cell death. Using the two-hybrid system in Saccharomyces cerevisiae and a system based on λ immunity in E.coli for in vivo detection of protein–protein interactions, we found that: (i) α protein interacts with Cnr, whereas α^cr proteins do not; (ii) both α–α and α^cr–α^cr interactions occur and the interaction domain is located within the C-terminal of α; (iii) Cnr–Cnr interaction also occurs. Using an in vivo competition assay, we found that Cnr interferes with both α–α and α^cr–α^cr dimerization. Our data suggest that Cnr and α interact in at least two ways, which may have different functional roles in P4 replication control.     

The decay of similarity over geographical distance in parasite communities of marine fishes

Aim We evaluate, for the first time, the decay of similarity over distance in four species of marine fishes with different vagility: Trachurus murphyi (pelagic, highly vagile), Merluccius gayi (demersal, highly vagile), Sebastes capensis (demersal, low vagility) and Hippoglossina macrops (benthic, non‐vagile). Location We use our own data of the species composition of parasite communities in four host species: T. murphyi (Perciformes), M. gayi (Gadiformes), H. macrops (Pleuronectiformes) and S. capensis (Scorpaeniformes), from the coast of Chile and southern Argentina. Trachurus murphyi, M. gayi and H. macrops in the area studied live closely associated with the cold waters of the Humboldt Current System, while S. capensis in southern Chile is associated with the fjord areas south of c. 43° S, and in Argentina this species lives in close association with the cold water of the Malvinas Current, which mixes with the warm water of the Brazilian Current in the sampled area. Thus, for all four host species there are no known geographical (oceanographic) barriers that interrupt their continuous distributions. Methods Fish were collected and frozen (?18 °C) until examination in the laboratory. After thawing, each visceral organ was dissected separately, washed in running water, filtered using a mesh of 0.25 mm, and examined under a stereomicroscope (20×). For S. capensis, only ectoparasites were included in this study because data on endoparasites for some locations are not available. The approach of Poulin [Journal of Biogeography 30 (2003) 1609] was used. For each host species, we computed similarity in parasite composition using the Bray–Curtis distance on a presence/absence matrix. Similarity between each pair of localities was correlated (linear regression on raw and transformed data) with the distance (in nautical miles) among those sites. Results Among conspecific populations of host fish, there was no correlation between sample size and parasite species richness. The same pattern was evident when all host populations were pooled and treated as a single sample. Except for H. macrops, an important proportion of the geographical ranges of the host species was covered. The decay in similarity of parasite communities over distance was recorded in three of the four hosts analysed. For H. macrops, no relationship between distances and decay of similarity was evident. A linear regression on non‐transformed data provided a good fit, and the regression on log(n + 1) does not change significantly for both r² and probability, but slopes are smaller for log‐transformed than for non‐transformed data. Main conclusions Three of the four fish parasite systems studied fit well with the expected decay of similarity over distance. The slopes of the relationship between distance and similarity in the marine fishes studied are lower, but significant (three orders of magnitude) than those recorded in freshwater fishes by Poulin (2003) . In our case, Hippoglossina macrops (benthic, non‐vagile host) does not show any relationship, suggesting homogeneity over the geographical area studied for this species.     

Metazoan parasites of Engraulis ringens as tools for stock discrimination along the Chilean coast

The value of the metazoan parasites of the anchoveta Engraulis ringens as biological tags for stock discrimination is assessed. Three hundred and fifty-nine specimens, obtained from six landing ports in northern and central Chile (Arica, Antofagasta, Caldera, Coquimbo, Valparaiso and Talcahuano) covering a latitudinal gradient of c . 18°, were analysed. Six metazoan parasite species (three ectoparasites and three endoparasites) were obtained. A correspondence analysis suggested that anchoveta from northern and central Chile are comprised of two stocks. Identification of stocks was largely based on qualitative differences (absence in southern localities of the monogenean Pseudanthocotyloides heterocotyle , a parasite proper of engraulids, and the copepod Caligus sp.) and quantitative differences in the prevalence of infection for the remaining species. Inertia and mass value for the correspondence analysis supported the hypothesis of two stocks.     

Photosynthetic limitations in leaves of young Brazilian Green Dwarf coconut (Cocos nucifera L. ‘nana’) palm under well-watered conditions or recovering from drought stress

Young plants of two Brazilian Green Dwarf coconut ecotypes from contrasting regions in relation to climate (UGD, from a hot and humid climate and JGD, from a hot and dry climate) were submitted to three consecutive drying/recovery cycles, under greenhouse conditions to determine the photosynthetic limitations encountered during the recovery phase of water deficiency. Three hypotheses were tested: (1) non-stomatal factors contribute to the incomplete recovery of net photosynthesis rate (A), (2) photochemical impairment is an important component of the non-stomatal limitations to A and (3) the two coconut ecotypes respond differently to the drought recovery. In each drying/recovery cycle, irrigation was suppressed until A reached zero. Then the plants were rewatered and the recovery was accompanied for 8 days. After 4 days of rewatering, CO₂ and light response curve parameters were used to discriminate the stomatal and non-stomatal factors contributing to the incomplete recovery of A. Upon rewatering for 4 days, the pre-dawn leaf water potential of the stressed plants increased from ?1.20 to ?0.15 MPa on average. Only small differences were detected in the intercellular to atmospheric CO₂ concentration ratio but A did not recover completely to the control values. In both ecotypes, light and CO₂-saturated A, carboxylation efficiency, maximum quantum yield of PSII and maximum electron transport rate were significantly lower in stressed plants in comparison with the well-watered ones. Relative stomatal limitation (L_S) ranged from 16 to 20% without significant differences between treatments and between ecotypes. The relative mesophyll limitation (L_m) was significantly higher in JGD in cycle 1 but this changed after repeated drought stress, decreasing to values significantly lower than those in UGD in the other two cycles. The results showed that after 4 days of rewatering stressed plant stomatal limitations were comparable to that of well-watered control plants (～20%). However A did not recover completely after drought stress due to non-stomatal factors. Important differences between the ecotypes were identified. The ecotype JGD presented smaller L_m, faster recovery of the apparent quantum efficiency of CO₂ assimilation, adjustment of some carbon balance components and signs of photosynthetic capacity acclimation after repeated drying/recovery cycles.     

Permanent genetic resources added to Molecular Ecology Resources Database 1 February 2011-31 March 2011

This article documents the addition of 111 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Acipenser oxyrinchus desotoi, Anopheles nuneztovari sensu lato, Asellus aquaticus, Calopteryx splendens, Calopteryx virgo, Centaurea aspera, Centaurea seridis, Chilina dombeyana, Proctoeces cf. lintoni and Pyrenophora teres f. teres.     

The Complete Amino Acid Sequence of a Trypsin Inhibitor from Bauhinia variegata var. candida Seeds

Trypsin inhibitors of two varieties of Bauhinia variegata seeds have been isolated and characterized. Bauhinia variegata candida trypsin inhibitor (BvcTI) and B. variegata lilac trypsin inhibitor (BvlTI) are proteins with M _r of about 20,000 without free sulfhydryl groups. Amino acid analysis shows a high content of aspartic acid, glutamic acid, serine, and glycine, and a low content of histidine, tyrosine, methionine, and lysine in both inhibitors. Isoelectric focusing for both varieties detected three isoforms (pI 4.85, 5.00, and 5.15), which were resolved by HPLC procedure. The trypsin inhibitors show K _i values of 6.9 and 1.2 nM for BvcTI and BvlTI, respectively. The N-terminal sequences of the three trypsin inhibitor isoforms from both varieties of Bauhinia variegata and the complete amino acid sequence of B. variegata var. candida L. trypsin inhibitor isoform 3 (BvcTI-3) are presented. The sequences have been determined by automated Edman degradation of the reduced and carboxymethylated proteins of the peptides resulting from Staphylococcus aureus protease and trypsin digestion. BvcTI-3 is composed of 167 residues and has a calculated molecular mass of 18,529. Homology studies with other trypsin inhibitors show that BvcTI-3 belongs to the Kunitz family. The putative active site encompasses Arg (63)–Ile (64).