Characterization and optimization of ArtinM lectin expression in Escherichia coli

Background ArtinM is a D-mannose-specific lectin from Artocarpus integrifolia seeds that induces neutrophil migration and activation, degranulation of mast cells, acceleration of wound healing, induction of interleukin-12 production by macrophages and dendritic cells, and protective T helper 1 immune response against Leishmania major, Leishmania amazonensis and Paracoccidioides brasiliensis infections. Considering the important biological properties of ArtinM and its therapeutic applicability, this study was designed to produce high-level expression of active recombinant ArtinM (rArtinM) in Escherichia coli system. Results The ArtinM coding region was inserted in pET29a(+) vector and expressed in E. coli BL21(DE3)-Codon Plus-RP. The conditions for overexpression of soluble ArtinM were optimized testing different parameters: temperatures (20, 25, 30 or 37°C) and shaking speeds (130, 200 or 220 rpm) during induction, concentrations of the induction agent IPTG (0.01-4 mM) and periods of induction (1-19 h). BL21-CodonPlus(DE3)-RP cells induced under the optimized conditions (incubation at 20°C, at a shaking speed of 130 rpm, induction with 0.4 mM IPTG for 19 h) resulted in the accumulation of large amounts of soluble rArtinM. The culture provided 22.4 mg/L of rArtinM, which activity was determined by its one-step purification through affinity chromatography on immobilized D-mannose and glycoarray analysis. Gel filtration showed that rArtinM is monomeric, contrasting with the tetrameric form of the plant native protein (jArtinM). The analysis of intact rArtinM by mass spectrometry revealed a 16,099.5 Da molecular mass, and the peptide mass fingerprint and esi-cid-ms/ms of amino acid sequences of peptides from a tryptic digest covered 41% of the total ArtinM amino acid sequence. In addition, circular dichroism and fluorescence spectroscopy of rArtinM indicated that its global fold comprises β-sheet structure. Conclusions Overall, the optimized process to express rArtinM in E. coli provided high amounts of soluble, correctly folded and active recombinant protein, compatible with large scale production of the lectin.

Contribuição para o estudo dos Rhinotragini (Coleoptera, Cerambycidae).VII. O gênero Ischasioides

Eclipta brasiliensis (Fisher, 1947) is transferred to Ischasioides Tavakilian & Peñaherrera-Leiva, 2003, and synonymized with I. crassitarsis (Gounelle, 1911). Ischasia m. atrocephala Fuchs, 1956 and Ischasia m. nigrovittata Fuchs, 1956, although without nomenclatural status, are confirmed as equal to I. crassitarsis. Ischasioides berkovae sp. nov. is described from Brazil. A key to the species of Ischasioides is provided.

Concentração e redistribuição de nutrientes minerais nos diferentes estádios foliares de seringueira

A seringueira na fase adulta possui um crescimento intermitente com o processo de troca de folhas, que é caracterizado pela senescência. Esse é um mecanismo que as árvores utilizam para reciclarem os nutrientes, por meio dos ciclos bioquímico e biogeoquímico. Estudos sobre o ciclo bioquímico são necessários para verificar o comportamento dos nutrientes minerais, nos diferentes estádios foliares, pois a conservação desses é importante, visto que os seringais são implantados em solos de baixa fertilidade. Desse modo, este trabalho teve por objetivo avaliar a dinâmica dos nutrientes minerais, durante os diferentes estádios foliares da seringueira. As folhas utilizadas foram provenientes de plantas do clone RRIM-600, de um seringal localizado em Nepomuceno, MG. Foram selecionadas oito árvores adultas que possuíam folhas nos diferentes estádios (B1, B2, C e D) e as senescentes (Sen.). As folhas nos estádios B1, B2, C e D foram coletadas nos quatro pontos ortogonais e as senescentes foram coletadas no solo após a agitação mecânica dos galhos. Observou-se que os nutrientes N, P, K, S, Cu e Zn diminuíram os seus teores nos diferentes estádios foliares da seringueira. No entanto, para Ca, Mg, B, Fe e Mn, verificarou-se um comportamento inverso. A redistribuição dos nutrientes foi maior para N, P, K e Cu. A ordem relativa dos teores de macronutrientes encontrados nas folhas foi: N>K>Ca>Mg>S>P e, para os micronutrientes, a ordem foi: Mn>Fe>Zn>B>Cu.

Potencial do flavedo (epicarpo) de Citrus aurantifolia cv. Tahiti no controle do bolor verde e da antracnose em citros

O Brasil é considerado o maior produtor de citros e o maior exportador de suco de laranja. Doenças de pós-colheita representam uma grande perda para a citricultura, sendo que para a exportação de frutos são rígidas as exigências com relação a isenção de resíduos químicos nos mesmos. Patógenos de importância em pós-colheita de citros incluem o Penicillium digitatum, agente causal do bolor-verde e o Colletotrichum gloeosporioides, agente causal da antracnose. Dada a importância econômica que representam estas doenças dos frutos cítricos, tanto em termos de comprometimento da qualidade e dificuldade de controle, a busca de alternativas adicionais que possam viabilizar a capacidade produtiva e garantir a obtenção de frutos com excelentes padrões de qualidade torna-se imprescindível. Portanto, estudou-se os efeitos dos extratos aquosos do flavedo de Citrus aurantifolia var. Tahiti, Lentinula edodes, Agaricus subrufescens (syn. Agaricus brasiliensis), albedo de Citrus sinensis var. Valência e do ácido jasmônico no controle póscolheita do bolor verde e da antracnose e na indução de resistência em frutos de laranjeira Valência (Citrus sinensis). Foi possível observar que o extrato aquoso do flavedo (C. aurantifolia) apresentou efeito inibitório sobre os patógenos, quando tratados em pós-colheita, em função da redução dos sintomas e esporulação. Porém, os extratos de albedo (C. sinensis), L. edodes, A. subrufescens e o ácido jasmônico não apresentaram efeitos sobre P. digitatum e C. gloeosporioides.

A retrospective study of oral manifestations in patients with paracoccidioidomycosis

South American blastomycosis, paracoccidioidomycosis (Pb mycosis) or Lutz disease is an endemically fungal infection in Latin America. It is caused by the dimorphic fungus Paracoccidioides brasiliensis and may cause oral mucosal lesions. The incidence of Pb mycosis oral lesions was evaluated in patients assisted at a Brazilian Dental School's Specialized Oral Diagnosis Service with special focus on the different clinical forms of these lesions, its location, patients' occupation, deleterious habits, and diagnosis methodology. Students' and professionals' initial diagnoses were compared with the definitive diagnosis. Lesions were detected 31 cases (18 patients). The results show that 88.8% of the patients were male with a mean age of 50 years and 39% work(ed) with activities related to agriculture. As much as 88.9% were smokers and 72.2% were alcohol users. Exfoliative cytology was performed in 66.6% of the patients. Oral mucosa (30%), gingiva (16.6%) and lips 16.6% were the most common sites of Pb mycosis oral lesions. Comparing the initial with the definitive diagnosis made by the professionals their accuracy was 33% (6 out of 18 patients). Students' diagnosis was more accurate demonstrating 72.5% of initial correct diagnosis (13 out of 18). Statistical analysis by ANOVA (α=0.05, SPSS WIN) demonstrated a significant difference between the diagnosis of Pb mycosis made by students and professionals when considering initial diagnosis and final diagnosis (after histopathological analysis) (p=0.25). Incisional biopsy and exfoliate cytology are efficient for an early diagnosis of this disease in mouth. Students' training in diagnosis of oral pathologies to recognize lesions is urgent to improve public health.

Range extensions along western Atlantic for Epialtidae crabs (Brachyura, Majoidea) genera Acanthonyx Latreille, 1828 and Epialtus H. Milne Edwards, 1834

The present study provided information extending the known geographical distribution of three species of majoid crabs, the epialtids Acanthonyx dissimulatus Coelho, 1993, Epialtus bituberculatus H. Milne Edwards, 1834, and E. brasiliensis Dana, 1852. Specimens of both genera from different carcinological collections were studied by comparing morphological characters. We provide new data that extends the geographical distributions of E. bituberculatus to the coast of the states of Paraná and Santa Catarina (Brazil), and offer new records from Belize and Costa Rica. Epialtus brasiliensis is recorded for the first time in the state of Rio Grande do Sul (Brazil), and A. dissimulatus is reported from Quintana Roo, Mexico. The distribution of A. dissimulatus, previously known as endemic to Brazil, has a gap between the states of Espírito Santo and Rio de Janeiro. However, this restricted southern distribution is herein amplified by the Mexican specimens.

Feeding habits of the atlantic spotted dolphin, Stenella frontalis, in southeastern Brazil

This study presents new information on the feeding habits of the Atlantic spotted dolphins, Stenella frontalis, in the Western South Atlantic. Nine stomach contents of S. frontalis incidentally caught in fishing operations conducted by the gillnet fleet based on main harbour of Cananéia (25°00'S; 47°55'W), southeastern Brazil, were analyzed. These specimens were captured between 2005 and 2007. A total of 1 422 cephalopod beaks, 147 otoliths and three crustaceans were recovered from the stomach contents. The dolphins assessed preyed on at least eight different fish species of the families Trichiuridae, Carangidae, Sparidae, Merluccidae, Engraulidae, Sciaenidae, Congridae and Scombridae, five cephalopod species of the families Loliginidae, Sepiolidae, Tremoctopodidae and Thysanoteuthidae, and one shrimp species of the Penaeidae family. Based on the analysis of the Index of Relative Importance (IRI), the Atlantic cutlassfish, Trichiurus lepturus, was the most important fish species represented. Of the cephalopods, the squid Doryteuthis plei was by far the most representative species. Several items were reported for the first time as prey of the S. frontalis: Xiphopenaeus kroyeri, Tremoctopus violaceus, Semirossia tenera, Merluccius hubbsi, Pagrus pagrus and Paralonchurus brasiliensis. S. frontalis presented teuthophagous and ichthyofagous feeding habits, with apparent predominance of the first, and preyed mainly on pelagic and demersal items.

Brazilian sandy beach macrofauna production: a review

The state of the art of the studies on the production of Brazilian sandy beach macrofauna was analyzed on the basis of the data available in the literature. For this purpose, the representativeness of the production dataset was examined by latitudinal distribution, degree of exposure and morphodynamic state of beaches, taxonomic groups, and methods employed. A descriptive analysis was, further, made to investigate the trends in production of the more representative taxonomic groups and species of sandy beach macrofauna. A total of 69 macrofauna annual production estimates were obtained for 38 populations from 25 studies carried out between 22º56'S and 32º20'S. Production estimates were restricted to populations on beaches located on the southern and southeastern Brazilian coast. Most of the populations in the dataset inhabit exposed dissipative sandy beaches and are mainly represented by mollusks and crustaceans, with a smaller number of polychaetes. The trends in production among taxonomic groups follow a similar pattern to that observed on beaches throughout the world, with high values for bivalves and decapods. The high turnover rate (P/B ratio) of the latter was due to the presence of several populations of the mole crab Emerita brasiliensis, which can attain high values of productivity, in the dataset. Most of the studies focus on the comparison of production and, especially, of P/B ratio according to life history traits in populations of the same species/taxonomic group. Despite the importance of life history-production studies, other approaches, such as the effect of man-induce disturbances on the macrofauna, should be undertaken in these threatened environments.

In pulmonary paracoccidioidomycosis IL-10 deficiency leads to increased immunity and regressive infection without enhancing tissue pathology

BACKGROUND: Cellular immunity is the main defense mechanism in paracoccidioidomycosis (PCM), the most important systemic mycosis in Latin America. Th1 immunity and IFN-γ activated macrophages are fundamental to immunoprotection that is antagonized by IL-10, an anti-inflammatory cytokine. Both in human and experimental PCM, several evidences indicate that the suppressive effect of IL-10 causes detrimental effects to infected hosts. Because direct studies have not been performed, this study was aimed to characterize the function of IL-10 in pulmonary PCM. METHODOLOGY/PRINCIPAL FINDINGS: Wild type (WT) and IL-10(-/-) C57BL/6 mice were used to characterize the role of IL-10 in the innate and adaptive immunity against Paracoccidioides brasiliensis (Pb) infection. We verified that Pb-infected peritoneal macrophages from IL-10(-/-) mice presented higher phagocytic and fungicidal activities than WT macrophages, and these activities were associated with elevated production of IFN-γ, TNF-α, nitric oxide (NO) and MCP-1. For in vivo studies, IL-10(-/-) and WT mice were i.t. infected with 1×10(6) Pb yeasts and studied at several post-infection periods. Compared to WT mice, IL-10(-/-) mice showed increased resistance to P. brasiliensis infection as determined by the progressive control of pulmonary fungal loads and total clearance of fungal cells from dissemination organs. This behavior was accompanied by enhanced delayed-type hypersensitivity reactions, precocious humoral immunity and controlled tissue pathology resulting in increased survival times. In addition, IL-10(-/-) mice developed precocious T cell immunity mediated by increased numbers of lung infiltrating effector/memory CD4(+) and CD8(+) T cells. The inflammatory reactions and the production of Th1/Th2/Th17 cytokines were reduced at late phases of infection, paralleling the regressive infection of IL-10(-/-) mice. CONCLUSIONS/SIGNIFICANCE: Our work demonstrates for the first time that IL-10 plays a detrimental effect to pulmonary PCM due to its suppressive effect on the innate and adaptive immunity resulting in progressive infection and precocious mortality of infected hosts.

Imported human rabies in Switzerland, 2012: a diagnostic conundrum

Human rabies is rare in Western Europe. It is not easily recognized in the absence of a history of exposure. We describe the clinical course, diagnosis and follow-up of an imported human rabies case in Switzerland. The patient, a U.S. citizen, presented at an outpatient clinic in Iraq with pain in his right shoulder on July 5, 2012. On July 8 he was transferred to a hospital in the United Arab Emirates, where he exhibited progressive encephalitis with coma. On July 29, he was transferred to a hospital in Switzerland, where he died on July 31, 2012. The autopsy showed severe encephalitis. Rabies was diagnosed by the rapid fluorescent focus inhibition test (RFFIT) and confirmed by fluorescence antibody testing (FAT) in brain smears and immunohistochemistry on paraffin-embedded brain sections. The viral strain was characterized by RT-PCR followed by sequencing and phylogenetic analysis as an American bat rabies strain associated with Tadarida brasiliensis. Close contacts and exposed health care workers received postexposure prophylaxis (PEP).

Mineralogy and nannoplankton distribution in ODP Hole 194-1195A

Significant synchronous shifts in the chemistry, mineralogy, grain sizes and color of the sediments at 6 m below sea floor (mbsf) at ODP Site 1195 on the Marion Plateau (NE Australia) are interpreted to reflect a major regional paleoceanographic change: the initiation of the southern province of the Great Barrier Reef (GBR). The onset of this massive carbonate production centre nearby resulted primarily in increased deposition of carbonate-rich sediments of neritic origin. Both sedimentation rate and terrigenous input record a coincident decline attributed to inshore trapping of materials behind the reefs. Our best estimate places the development of reef framework in the southern part of the GBR between 560 and 670 kyr B.P., based on an age model combining magnetostratigraphic and biostratigraphic data. The proposed estimation agrees with previous studies reporting an age between 500 and 930 kyr B.P., constraining more tightly their results. However, it does not support research placing the birth of the GBR in Marine Isotope Stage (MIS) 11 (~400 kyr), nor the theory of a worldwide modern barrier reef development at that time.

Ostracoda fauna of surface sediments from the Persian Gulf

During the "Meteor"-Expedition to the Persian Gulf in March-May 1965, approximately 300 samples were collected. Most of them have been already studied by various authors in sedimentological as well as micropaleontological respects. 49 samples were selected for ostracode studies. These samples are arranged to form a long-axis section ("Laengsprofil"), and 4 shorter cross-profiles, perpendicular to the long-axis profile in the Persian Gulf and Gulf of Oman. 52 species of ostracodes in this area were specifically determined; 39 of them are described under open nomenclature. 13 species are already known from surrounding sea areas: 2 species from the Red Sea; 2 species from the east coast of Africa; 1 species from the Mediterranean Sea; and others from the Indian and Pacific Oceans. 12 species show close relationships to species from the Indopacific Ocean. The ostracode species found in the area can be grouped after the method of BRAUN-BLANQUT into 2 bioassociations. Association 1 with the following 4 characteristic species : Cytherella cf. pulchra, Loxoconcha sp. A, Neomonoceratina sp. A, Alocopocythere reticulata. Association 2 with 1 characteristic species: Ruggieria (Ruggieria) sp. B. The association 1 is widespread in the entire studied area of the Persian Gulf, where it is considered to characterize the shallow water region down to 200 m. The association 2 is restricted to the deeper water below 200 m of the inner part of the Oman Gulf. Only a few species known from the shallow water association of the Persian Gulf are present. Within the two ostracode associations mentioned above 4 zones from the total studied area could be related to the water depth. The zones A-D are characterized more or less readily by the relative abundance of certain species: Zone A : 7-30 m depth, on substrates of poorly coarse-grained clayey marl; Zone B: 30-94 m depth, on substrates of richly coarse-grained calcareous marl; Zone C: 94-1961208 m depth, on substrates of richly coarse-grained calcareous marl; Zone D: 196/208-500 m depth, on substrates of calcareous clay, poor in benthos. The regional and bathymetric distribution of the ostracode fauna in the area studied was compared in relation to 10 environmental factors: water depth, temperature, salinity, water density, O2-concentration, phosphate-silica contents, pH-values, stratification of the water body, water currents and type of sediments. The major environmental factors which appear to control the ostracode distribution are water depth (as a complex factor), O2-concentration and the type of sediment. At 3 stations (GIK01058, GIK01074 and GIK01204) species of the shallow water association were found together with a few bathyal species. These stations are situated at the outer Biaban shelf, in an area where the bottom water of the Persian Gulf flows down the slope towards the Oman Gulf. Several samples of the Zone B in the major part of the Persian Gulf show also a high species diversity containing a high percentage of subfossil ostracode carapaces. It is probable that the recent biocoenosis has been mixed with a late quarternary thanatocoenosis.

The Global Plankton Database: an inventory and data from the Former Soviet Union expeditions

At present time, there is a lack of knowledge on the interannual climate-related variability of zooplankton communities of the tropical Atlantic, central Mediterranean Sea, Caspian Sea, and Aral Sea, due to the absence of appropriate databases. In the mid latitudes, the North Atlantic Oscillation (NAO) is the dominant mode of atmospheric fluctuations over eastern North America, the northern Atlantic Ocean and Europe. Therefore, one of the issues that need to be addressed through data synthesis is the evaluation of interannual patterns in species abundance and species diversity over these regions in regard to the NAO. The database has been used to investigate the ecological role of the NAO in interannual variations of mesozooplankton abundance and biomass along the zonal array of the NAO influence. Basic approach to the proposed research involved: (1) development of co-operation between experts and data holders in Ukraine, Russia, Kazakhstan, Azerbaijan, UK, and USA to rescue and compile the oceanographic data sets and release them on CD-ROM, (2) organization and compilation of a database based on FSU cruises to the above regions, (3) analysis of the basin-scale interannual variability of the zooplankton species abundance, biomass, and species diversity.