Secretion of an endogenous subtilisin by Pichia pastoris strains GS115 and KM71.

The methylotrophic yeast Pichia pastoris is widely used for the expression of heterologous enzymes. While the purity of the desired expression product is of major importance for many applications, we found that recombinant enzymes produced in methanol medium were contaminated by a 37-kDa endogenous yeast protease. This enzyme was completely inhibited by phenylmethanesulfonyl fluoride (PMSF) but not by 1,10-phenanthroline, EDTA, and pepstatin A, suggesting the nature of a serine protease. Its secretion was abolished in P. pastoris strains GS115 and KM71 by specific mutagenesis of a subtilisin gene (SUB2) but not by inactivation of the gene encoding vacuolar proteinase B (PRB). Bioinformatic comparisons of Sub2 protein with subtilisins from other fungal genomes and phylogenetic analyses indicated that this enzyme is not an orthologue of the vacuolar protease cerevisin generally present in yeasts but is more closely related to another putative subtilisin found in a small number of yeast genomes. During growth of P. pastoris, Sub2 was produced as a secreted enzyme at a concentration of 10 microg/ml of culture supernatant after overexpression of the full-length SUB2 gene. During fermentative production of recombinant enzymes in methanol medium, 1 ml of P. pastoris culture supernatant was found to contain approximately 3 ng of Sub2, while the enzyme was not detected during growth in a medium containing glycerol as a carbon source. The mutant strain GS115-sub2 was subsequently used as a host for the production of recombinant proteases without endogenous subtilisin contamination.

Description of Photuris fulvipes (Blanchard) immatures (Coleoptera, Lampyridae, Photurinae) and bionomic aspects under laboratory conditions

Larvae and pupae of the cryptic firefly Photuris fulvipes (Blanchard,1837) are described. They were reared to adult stage from eggs laid by a female collected in São Luiz do Paraitinga, São Paulo, Brazil. Additionally, adult diagnosis and some bionomic data observed in the laboratory are presented. The bionomical and morphological characteristics of P. fulvipes were compared to other unidentified species of the genus and to Bycellonycha sp. from literature. At moment, the data are insufficient to propose a diagnosis for the species, but some characteristics appear to define species group or, at least, they may have generic value.

Vaccine potentials of an intrinsically unstructured fragment derived from the blood stage-associated Plasmodium falciparum protein PFF0165c.

We have identified new malaria vaccine candidates through the combination of bioinformatics prediction of stable protein domains in the Plasmodium falciparum genome, chemical synthesis of polypeptides, in vitro biological functional assays, and association of an antigen-specific antibody response with protection against clinical malaria. Within the predicted open reading frame of P. falciparum hypothetical protein PFF0165c, several segments with low hydrophobic amino acid content, which are likely to be intrinsically unstructured, were identified. The synthetic peptide corresponding to one such segment (P27A) was well recognized by sera and peripheral blood mononuclear cells of adults living in different regions where malaria is endemic. High antibody titers were induced in different strains of mice and in rabbits immunized with the polypeptide formulated with different adjuvants. These antibodies recognized native epitopes in P. falciparum-infected erythrocytes, formed distinct bands in Western blots, and were inhibitory in an in vitro antibody-dependent cellular inhibition parasite-growth assay. The immunological properties of P27A, together with its low polymorphism and association with clinical protection from malaria in humans, warrant its further development as a malaria vaccine candidate.

The adequacy of artificial oviposition substrates for laboratory rearing of Piezodorus guildinii (Westwood) (Heteroptera, Pentatomidae)

Studies in the laboratory tested the suitability of synthetic wool string, cotton string, cheesecloth, and commercial cotton ball as artificial oviposition substrates for the small green stink bug, Piezodorus guildinii (Westwood) (Heteroptera, Pentatomidae). In confined cages, 54% of total egg masses was laid on synthetic wool string, 31% on cotton string, and only 15% on cheesecloth. In an additional test, the best substrate selected, synthetic wool string, received 92% of egg masses compared to 8% on the commonly used substrate, cotton ball. Synthetic wool string received the most egg masses of any size, in particular those in the range 11-20 eggs/mass. Because the eggs of P. guildinii are laid in two parallel double rows, the egg masses fit the wool string perfectly.

A new species of the genus Pimelerodius Vanin from the Amazon Region, with notes on the geografic distribution of Pimelerodius motacilla (Boheman) (Coleoptera, Curculionidae, Otidocephalini)

Pimelerodius punctiventris sp. nov. (type locality Brazil, Amazonas, Itacoatiara) is described and illustrated. The new taxon is compared with similar species, being distinguished from the other 12 known species of the genus by the presence of punctures in ventrite I. The available published key for identification of species of Pimelerodius is adapted to include the new species. A modification of the generic description of the aedeagus of Pimelerodius is provided, a necessity due to the differences observed in the aedeagus of the new species. The occurrence of P. motacilla (Boheman, 1843) in the Amazon Region, recorded in sympatry with P. punctiventris in Itacoatiara, AM, is discussed and confirmed, based on the study of 41 available specimens.

Morphological and physiological variation between queens and workers of Protonectarina sylveirae (de Saussure) (Hymenoptera, Vespidae, Epiponini)

Morphological and physiological variation between queens and workers of Protonectarina sylveirae (de Saussure) (Hymenoptera, Vespidae, Epiponini). The Neotropical swarm-founding wasps, Epiponini, range from the absence of morphological differentiation between castes to highly distinct castes. We measured eight body parts of females of two colonies of Protonectarina sylveirae (de Saussure, 1854). ANOVA and Discriminant Analysis evidenced significant differences between castes, as previously observed by other authors for other species of Epiponini. However, some females previously categorized as queens, were actually workers, supported by our statistic analyses. These individuals showed intermediate morphological features between queens and workers, having distinct patterns of hairs and clypeal spots. The castes of P. sylveirae are distinct, however intermediate individuals may be found in colonies promoting social flexibility.

On p-beauty contest integer games

In this paper we provide a full characterization of the pure-strategyNash Equilibria for the p-Beauty Contest Game when we restrict player schoices to integer numbers. Opposed to the case of real number choices,equilibrium uniqueness may be lost depending on the value of p and thenumber of players: in particular, as p approaches 1 any symmetric profileconstitutes a Nash Equilibrium. We also show that any experimental p-BeautyContest Game can be associated to a game with the integer restriction andthus multiplicity of equilibria becomes an issue. Finally, we show thatin these games the iterated deletion of weakly dominated strategies maynot lead to a single outcome while the iterated best-reply process alwaysdoes (though the outcome obtained depends on the initial conditions).

Systematic Assessment of Factors Influencing Preferences of Crohn's Disease Patients in Selecting an Anti-TNF agent (CHOOSE TNF TRIAL)

Background a nd A ims: I nfliximab (IFX), adalimumab (ADA)and certolizumab pegol (CZP) have similar efficacy for inductionand maintenance of clinical response and remission in Crohn'sdisease (CD). Given the comparable nature of t hese drugs,patients' p references m ay i nfluence the choice o f the product.Goal: to identify factors contributing to CD patients' decision inselecting one anti-TNF agent over the others.Methods: A p rospectdive s urvey was performed a mong a nti-TNF-naïve CD patients. Prior to completion of a questionnaire,patients were provided with a description of the three anti-TNFagents f ocusing on indications, route of administration, s ideeffects, and scientific evidence of efficacy and safety.Results: One hundred patients (47f/53m, mean age 45±16yrs)completed the questionnaire. Disease location was ileal, colonicand ileocolonic in 33%, 40% and 27% of patients, respectively.Thirty-six percent preferred ADA as medication of choice, while28% and 2 5% p referred CZP and IFX; 11% were u ndecided.Patients' decision in selecting an anti-TNF drug was influencedby t he following f actors: side effects ( 76%), p hysician'srecommendation (66%), route of administration (54%), efficacydata (52%), time required for therapy administration (27%),recommendations by other CD patients (21%) and interactionswith other medications (12%).Conclusions: T he majority of p atients p referred anti-TNFmedications t hat were a dministered by s ubcutaneous i njectionrather t han b y intravenous i nfusion. Side effect profile andphysicians' r ecommendation are t wo m ajor factors influencingthe patients' s election of a specific anti-TNF d rug. Patients'concerns about safety and lifestyle habits should be taken intoaccount when prescribing anti-TNF drugs.

Analysis of digestive tract content of the larvae of Polybia scutellaris (White) (Hymenoptera, Vespidae)

Polybia scutellaris (White, 1841) is a social wasp of biological interest for its role as pollinator and maybe as biological control agent of sanitary and agricultural pests. This study examines the digestive tract contents of the larvae of P. scutellaris from four nests in Magdalena (Buenos Aires province, Argentina). Contents included both animal (arthropod parts) and plant (pollen, leaf and fruit epidermis) parts. The pollen content analysis showed that the wasps visited 19 different taxa of plants during the last active period of the colony before the nests had been collected. The range of sources used by P. scutellaris allows us characterizing the species as a generalist flower visitor. Wasps visited both native and exotic plants located nearby the nest. Most of the epidermal plant remains found in the larval digestive tract belonged to Malvaceae, a family not exploited by the studied colonies as pollen source.

Effect of gossypol on survival and reproduction of the zoophytophagous stinkbug Podisus nigrispinus (Dallas)

Effect of gossypol on survival and reproduction of the zoophytophagous stinkbug Podisus nigrispinus (Dallas). Gossypol is a sesquiterpene aldehyde found in cotton plants conferring resistance against herbivory. Although the effect of this sesquiterpenoid on insect pests of cotton is known, the interaction of this compound with zoophytophagous predators such as Podisus nigrispinus (Dallas) (Hemiptera, Pentatomidae) has not been studied so far. Thus, the objective of this study was to evaluate the effect of the purified gossypol on nymphs and adults of P. nigrispinus. Nymphs and adults of this predator were fed on Tenebrio molitor pupae and supplemented with solutions of gossypol at concentrations of 0.00, 0.05, 0.10, and 0.20% (w/v) during the nymphal and adult stages or, only during the adult stage of P. nigrispinus. The nymphal stage of the predator was, on average, two days longer when suplemmented with gossypol. Emerged adults had lower fecundity and egg hatching, especially at the highest gossypol concentration (0.20%) ingested during the nymphal and adult stages. However, this predator was not affected when it ingested the compound only during the adult stage. P. nigrispinus can have delayed nymphal development and lower reproductive performance when ingesting the gossypol during the nymphal and adult stages, but only at higher concentrations of gossypol than that produced by cotton plants.

Mortality of Plutella xylostella (Lepidoptera, Plutellidae) by parasitoids in the Province of Santa Fe, Argentina

Mortality of Plutella xylostella (Lepidoptera, Plutellidae) by parasitoids in the Province of Santa Fe, Argentina. Plutella xylostella (Linnaeus, 1758) (Lepidoptera, Plutellidae) larvae cause severe economic damage on cabbage, Brassica oleracea L. variety capitata (Brassicaceae), in the horticultural fields in the Province of Santa Fe, Argentina. Overuse of broad spectrum insecticides affects the action of natural enemies of this insect on cabbage. The objectives of this work were to identify the parasitoids of P. xylostella and to determine their influence on larva and pupa mortality. Weekly collections of larvae and pupae were randomly conducted in cabbage crops during spring 2006 and 2007. The immature forms collected were classified according to their developmental stage: L1 and L2 (Ls = small larvae), L3 (Lm = medium larvae), L4 (Ll = large larvae), pre-pupae and pupae (P). Each individual was observed daily in the laboratory until the adult pest or parasitoid emergence. We identified parasitoids, the number of instar and the percentage of mortality of P. xylostella for each species of parasitoid. Parasitoids recorded were: Diadegma insulare (Cresson, 1875) (Hymenoptera, Ichneumonidae), Oomyzus sokolowskii (Kurdjumov, 1912) (Hymenoptera, Eulophidae), Cotesia plutellae (Kurdjumov, 1912) (Hymenoptera, Braconidae) and an unidentified species of Chalcididae (Hymenoptera). Besides parasitoids, an unidentified entomopathogenic fungus was also recorded in 2006 and 2007. In 2006, the most successful parasitoids were D. insulare and O. sokolowskii, while in 2007 only D. insulare exerted a satisfactory control and it attacked the early instars of the pest.

Pineus boerneri Annand, 1928 (Hemiptera, Adelgidae) - a new species to Brazil: morphology of eggs, nymphs and adults

Pineus boerneri Annand, 1928 (Hemiptera, Adelgidae) _ a new species to Brazil: morphology of eggs, nymphs and adults. Pineus boerneri represents the first Adelgidae species recorded in Brazil. This aphid species forms extensive colonies on branches and trunk of Pinus spp., with apterous oviparous females, eggs and nymphs covered with white wax. The aim of this research is to study the morphology of eggs, nymphs, and adults to provide useful data for species identification in order to solve taxonomic issues. The study was based on morphometric data and optical and scanning electron microscopy images. First instar nymphs are active and can be easily distinguished from the others by their elongate minute yellowish body; well developed legs bearing a pair of sensorial setae at the apex of the second tarsomere; and antenna with three segments with a large rhinarium and distinct apical setae on the last segment. From the second to the fourth instar, nymphs are sessile, with round red body; they loose the third antennal segment and its sensorial structures, as well as the setae on the second tarsomere. The oviparous female is reddish-brown, with round body with about 0.76 mm diameter; legs reduced; antennae one-segmented; ovipositor distinct; numerous wax glands are present, mainly on the head. Accurate characterization of the species and distinction of the nymphal instars of P. boerneri were made possible by canonical analysis of morphometric data and morphological characters.

Genetic structure of populations of Pissodes castaneus (De Geer) (Coleoptera, Curculionidae) using amplified fragment length polymorphism

Genetic structure of populations of Pissodes castaneus (De Geer) (Coleoptera, Curculionidae) using amplified fragment length polymorphism. The objective of this study was to determine the genetic structure of populations of Pissodes castaneus from different areas and on different species of Pinus using the PCR-AFLP technique. Twenty samples were analyzed, representing 19 populations from Brazil and one from Florence, Italy, which is the region of origin of P. castaneus. The four combinations of primers generated a total of 367 fragments of DNA, and 100% of polymorphic loci, indicating high degree of molecular polymorphism. The dendrogram did not reveal trends for grouping the populations in relation to origin. The low genetic similarity (0.11 between the most distant groups) and genetic distances of 0.13 and 0.44 for 10 out of the 20 samples may indicate several founding events or multiple introductions of heterogeneous strains into Brazil. The allelic fixation index (Fst) was 0.3851, considered high, and the number of migrants (Nm) was 0.3991, indicating low gene flow among populations. The highest genetic distances were between the population from Irani, SC and Cambará do Sul, RS and Bituruna, PR, indicating an independent founding event or a particular allelic fixation in the former location. The high genetic diversity among populations points out that the populations are genetically heterogeneous with a diverse gene pool in the surveyed areas, what makes them to respond differently to control measures.

The type-species of Psilochlorops Duda (Diptera, Chloropidae) and its position in the phylogeny of the genus, with the description of a new species

The type-species of Psilochlorops Duda (Diptera, Chloropidae) and its position in the phylogeny of the genus, with the description of a new species. The genus Psilochlorops is known only for the Neotropical Region and had six described species to date. Psilochlorops niger sp. nov. is herein described and the male genitalia of P. clavitibia, the type-species of the genus, is described in detail. A new cladistic analysis of Psilochlorops is presented, including all known species of the genus.

Seasonality of Pelecinus polyturator (Drury) (Hymenoptera, Pelecinidae) in the Atlantic Rainforest of São Paulo State, Brazil

Seasonality of Pelecinus polyturator (Drury) (Hymenoptera, Pelecinidae) in the Atlantic Rainforest of São Paulo State, Brazil. A survey of the parasitoid wasp Pelecinus polyturator (Drury, 1773) (Hymenoptera, Pelecinidae) was carried out with five Malaise traps/area in five areas in the Atlantic Rainforest of São Paulo State, Brazil, between November 2009 and October 2010. The sampling effort in each locality amounted to 1,825 trap-days. Data were obtained from a total of 317 exemplars of P. polyturator, corresponding to 108 females and 209 males. The average sex ratio of the studied population was 0.52. The highest occurrence of P. polyturator was observed between November and March with frequency peak in January; about 95% of the specimens studied were captured at altitudes close to 1,000 m above sea level.