Characterization of new Schistosoma mansoni microsatellite loci in sequences obtained from public DNA databases and microsatellite enriched genomic libraries

In the last decade microsatellites have become one of the most useful genetic markers used in a large number of organisms due to their abundance and high level of polymorphism. Microsatellites have been used for individual identification, paternity tests, forensic studies and population genetics. Data on microsatellite abundance comes preferentially from microsatellite enriched libraries and DNA sequence databases. We have conducted a search in GenBank of more than 16,000 Schistosoma mansoni ESTs and 42,000 BAC sequences. In addition, we obtained 300 sequences from CA and AT microsatellite enriched genomic libraries. The sequences were searched for simple repeats using the RepeatMasker software. Of 16,022 ESTs, we detected 481 (3%) sequences that contained 622 microsatellites (434 perfect, 164 imperfect and 24 compounds). Of the 481 ESTs, 194 were grouped in 63 clusters containing 2 to 15 ESTs per cluster. Polymorphisms were observed in 16 clusters. The 287 remaining ESTs were orphan sequences. Of the 42,017 BAC end sequences, 1,598 (3.8%) contained microsatellites (2,335 perfect, 287 imperfect and 79 compounds). The 1,598 BAC end sequences 80 were grouped into 17 clusters containing 3 to 17 BAC end sequences per cluster. Microsatellites were present in 67 out of 300 sequences from microsatellite enriched libraries (55 perfect, 38 imperfect and 15 compounds). From all of the observed loci 55 were selected for having the longest perfect repeats and flanking regions that allowed the design of primers for PCR amplification. Additionally we describe two new polymorphic microsatellite loci.

Relationships of new world Phlebotomine sand flies (Diptera: Psychodidae) based on fossil evidence

The fossil record and systematics of phlebotomid sand flies, vectors of leishmaniasis and arbovirus in several regions of the world, strongly support that living genera existed long before the Oligocene (38 million years, myr). A common Phlebotominae ancestor was present in the Triassic period before the separations of continents (248 myr).

The use of biodiversity as source of new chemical entities against defined molecular targets for treatment of malaria, tuberculosis, and T-cell mediated diseases: a review

The modern approach to the development of new chemical entities against complex diseases, especially the neglected endemic diseases such as tuberculosis and malaria, is based on the use of defined molecular targets. Among the advantages, this approach allows (i) the search and identification of lead compounds with defined molecular mechanisms against a defined target (e.g. enzymes from defined pathways), (ii) the analysis of a great number of compounds with a favorable cost/benefit ratio, (iii) the development even in the initial stages of compounds with selective toxicity (the fundamental principle of chemotherapy), (iv) the evaluation of plant extracts as well as of pure substances. The current use of such technology, unfortunately, is concentrated in developed countries, especially in the big pharma. This fact contributes in a significant way to hamper the development of innovative new compounds to treat neglected diseases. The large biodiversity within the territory of Brazil puts the country in a strategic position to develop the rational and sustained exploration of new metabolites of therapeutic value. The extension of the country covers a wide range of climates, soil types, and altitudes, providing a unique set of selective pressures for the adaptation of plant life in these scenarios. Chemical diversity is also driven by these forces, in an attempt to best fit the plant communities to the particular abiotic stresses, fauna, and microbes that co-exist with them. Certain areas of vegetation (Amazonian Forest, Atlantic Forest, Araucaria Forest, Cerrado-Brazilian Savanna, and Caatinga) are rich in species and types of environments to be used to search for natural compounds active against tuberculosis, malaria, and chronic-degenerative diseases. The present review describes some strategies to search for natural compounds, whose choice can be based on ethnobotanical and chemotaxonomical studies, and screen for their ability to bind to immobilized drug targets and to inhibit their activities. Molecular cloning, gene knockout, protein expression and purification, N-terminal sequencing, and mass spectrometry are the methods of choice to provide homogeneous drug targets for immobilization by optimized chemical reactions. Plant extract preparations, fractionation of promising plant extracts, propagation protocols and definition of in planta studies to maximize product yield of plant species producing active compounds have to be performed to provide a continuing supply of bioactive materials. Chemical characterization of natural compounds, determination of mode of action by kinetics and other spectroscopic methods (MS, X-ray, NMR), as well as in vitro and in vivo biological assays, chemical derivatization, and structure-activity relationships have to be carried out to provide a thorough knowledge on which to base the search for natural compounds or their derivatives with biological activity.

Comparison of two commercial formulations of Bacillus thuringiensis var. israelensis for the control of Anopheles aquasalis (Diptera: Culicidae) at three salt concentrations

Anopheles aquasalis larvae are salt water tolerant, preferring concentrations between 10 and 20 parts per thousand (ppt). The larvicidal efficacy of two formulations of Bacillus thuringiensis var. israelensis (Vectobac-12AS® and Bactivec®), was investigated against An. aquasalis at salinities of 0, 10, and 20 ppt. A probit analysis was used to calculate the lethal concentrations (LC50 and LC95) for each product at each salinity. The LC50 and LC95 were higher for Bactivec® than Vectobac-12AS®, and for Bactivec®, the LC50 and LC95 increased with salinity. Vectobac-12AS® should thus be preferred to Bactivec® for An. aquasalis control, especially in saline breeding habitats.

Synthesis of limonene β-amino alcohol derivatives in support of new antileishmanial therapies

A series of seven limonene β-amino alcohol derivatives has been regioselectively synthesised in moderate to good yields. Two of these compounds were found to be significantly effective against in vitro cultures of the Leishmania (Viannia) braziliensis promastigote form in the micromolar range. The activities found for 3b and 3f were about 100-fold more potent than the standard drug, Pentamidine, in the same test, while limonene did not display any activity. This is the first report of antileishmanial activity by limonene β-amino alcohol derivatives.

Sand flies of Nicaragua: a checklist and reports of new collections

Sand flies within the genus Lutzomyia serve as the vectors for all species of the protozoan parasite Leishmania in the New World. In this paper, we present a summary of the 29 species of Lutzomyia and one of Brumptomyia previously reported for Nicaragua and report results of our recent collections of 565 sand flies at eight localities in the country from 2001-2006. Lutzomyia longipalpis was the predominant species collected within the Pacific plains region of western Nicaragua, while Lutzomyia cruciata or Lutzomyia barrettoi majuscula were the species most frequently collected in the central highlands and Atlantic plains regions. The collection of Lutzomyia durani (Vargas & Nájera) at San Jacinto in July 2001 is a new record for Nicaragua. Leishmaniasis is endemic to Nicaragua and occurs in three forms: cutaneous, mucocutaneous and visceral leishmaniasis. Cutaneous infections are the most prevalent type of leishmaniasis in Nicaragua and they occur in two different clinical manifestations, typical cutaneous leishmaniasis and atypical cutaneous leishmaniasis, depending on the species of the infecting Leishmania parasite. The distribution of sand flies collected during this study in relation to the geographic distribution of clinical forms of leishmaniasis in the country is also discussed.

In vitro evaluation of new terpenoid derivatives against Leishmania infantum and Leishmania braziliensis

The activity of five (1-5) abietane phenol derivatives against Leishmania infantum and Leishmania braziliensis was studied using promastigotes and axenic and intracellular amastigotes. Infectivity and cytotoxicity tests were performed with J774.2 macrophage cells using Glucantime as a reference drug. The mechanisms of action were analysed by performing metabolite excretion and transmission electron microscopy ultrastructural studies. Compounds 1-5 were more active and less toxic than Glucantime. The infection rates and mean number of parasites per cell observed in amastigote experiments showed that derivatives 2, 4 and 5 were the most effective against both L. infantum and L. braziliensis. The ultrastructural changes observed in the treated promastigote forms confirmed that the greatest cell damage was caused by the most active compound (4). Only compound 5 caused changes in the nature and amounts of catabolites excreted by the parasites, as measured by ¹H nuclear magnetic resonance spectroscopy. All of the assayed compounds were active against the two Leishmania species in vitro and were less toxic in mammalian cells than the reference drug.

Description of a new genus and new species of New World Phlebotominae (Diptera, Psychodidae)

A new genus and new species of Phlebotominae, Edentomyia piauiensis (Diptera, Psychodidae) from a cave in Piauí State, Brazil, are described. This new genus belongs to Phlebotomini, but its inclusion in any subdivision of this tribe depends on further study.

Hybotinae (Diptera, Empidoidea, Hybotidae) from the Dominican Republic: new records and description of new species

Hybotinae is being recorded for the first time to Dominican Republic where the subfamily becomes represented by the genera Euhybus Coquillett, 1895, Neohybos Ale-Rocha & Carvalho, 2003 and Syneches Walker, 1852. Euhybus and Neohybos occur exclusively in the Americas, and Syneches is cosmopolitan. Six new species are described and illustrated: Euhybus martiniensis sp. nov., Neohybos cinereus sp. nov., N. longicornis sp. nov., N. pruinosus sp. nov., N. setosus sp. nov. and Syneches dominicanus sp. nov.. Identification keys are provided for the species of the three hybotine genera registered in West Indies. The following species are recorded for the first time from the Dominican Republic: Syneches inversus Curran, 1928, S. pallidus Wilder, 1974 and S. vineus Wilder, 1974. The previously unknown female of S. inversus is described.

Descriptions of new species with a key to identification of the genus Neodexiopsis Malloch (Diptera, Muscidae) in Brazil

Nine new species of Neodexiopsis Malloch from Paraná, southern Brazil, are described: Neodexiopsis cinerea sp. nov. and N. paranaensis sp. nov. from Ponta Grossa; N. facilis sp. nov., N. legitima sp. nov., N. similis sp. nov. and N. uber sp. nov. from Guarapuava; N. erecta sp. nov., N. pura sp. nov. and N. rara sp. nov. from Colombo. A key to the identification of the Brazilian species of Neodexiopsis is also presented.

The genus Lycoderides Sakakibara, stat. nov., its composition and descriptions of new species (Hemiptera, Membracidae, Stegaspidinae)

The genus Lycoderides Sakakibara, stat. nov. , its composition and descriptions of new species (Hemiptera, Membracidae, Stegaspidinae).The subgenus Lycoderes (Lycoderides) Sakakibara, 1972 is raised to the genus category - Lycoderides stat. nov.. - : and it now includes: Lycoderides amazonicus (Sakakibara, 1991), comb. nov. , Lycoderides brevilobus (Sakakibara, 1972), comb. nov. , Lycoderides burmeisteri (Fairmaire, 1846), comb. nov. , Lycoderides cultratus (Sakakibara, 1991), comb. nov. , Lycoderides fernandezi (Strümpel, 1988), comb. nov. , Lycoderides fuscus (Amyot & Serville, 1843), comb. nov. , Lycoderides gradatus (Sakakibara, 1972), comb. nov. , Lycoderides hippocampus (Fabricius, 1803), comb. nov. , Lycoderides luteus (Funkhouser, 1940), comb. nov. , Lycoderides marginalis (Walker, 1851), comb. nov. , Lycoderides nathanieli (Cryan, 1999), comb. nov. , Lycoderides obtusus (Sakakibara, 1991), comb. nov. , Lycoderides pennyi (Sakakibara, 1991), comb. nov. , Lycoderides phasianus (Fowler, 1896), comb. nov. (= Enchenopa minamen Buckton, 1901,SYN. NOV: ), Lycoderides protensus (Sakakibara, 1991), comb. nov. , Lycoderides serraticornis (Fowler, 1896), comb. nov. , and Lycoderides strumpeli (Sakakibara, 1991), comb. nov. The following new species are described: Lycoderides abditus, sp. nov. , Lycoderides brulei,SP. NOV. (: both from French Guiana), Lycoderides capixaba, sp. nov. (from Brazil, Espírito Santo), Lycoderides cavichiolii, sp. nov. (from Brazil, Rio de Janeiro), Lycoderides meloi, sp. nov. (from Brazil, Bahia), and Lycoderides oliviae, sp. nov. (from Brazil, Minas Gerais). Other nomenclatural change: Stegaspis bracteata (Fabricius, 1787) = Lycoderes capitata Buckton, 1903, syn. nov. New records of geographical distribution and a key to the species are provided.

Field performance of new cowpea cultivars inoculated with efficient nitrogen-fixing rhizobial strains in the Brazilian Semiarid

The objective of this work was to evaluate the contribution of efficient nitrogen-fixing rhizobial strains to grain yield of new cowpea cultivars, indicated for cultivation in the Brazilian Semiarid region, in the sub-medium of the São Francisco River Valley. Two experiments were set up at the irrigated perimeters of Mandacaru (Juazeiro, state of Bahia) and Bebedouro (Petrolina, state of Pernambuco). The treatments consisted of single inoculation of five rhizobial strains - BR 3267, BR 3262, INPA 03-11B, UFLA 03-84 (Bradyrhizobiumsp.), and BR 3299T(Microvirga vignae) -, besides a treatment with nitrogen and a control without inoculation or N application. The following cowpea cultivars were evaluated: BRS Pujante, BRS Tapaihum, BRS Carijó, and BRS Acauã. A randomized complete block design, with four replicates, was used. Inoculated plants showed similar grain yield to the one observed with plants fertilized with 80 kg ha-1 N. The cultivars BRS Tapaihum and BRS Pujante stood out in grain yield and protein contents when inoculated, showing their potential for cultivation in the sub-medium of the São Francisco River Valley.

Synthesis and characterization of new amino acyl-4-thiazolidones

A series of heterocyclic compounds with a 4-thiazolidone nucleus and amino acyl moiety were synthesized by protection reaction of thiosemicarbazide using the symmetrical anhydride (Boc)2O and cyclization with chloroacetic acid under mild conditions. Trifluoroacetic acid was used to obtain 4-thiazolidone and the alpha-amino acid condensation reactions were carried out using strategies for peptide synthesis. The characterization of this new class of compounds was performed using IR and ¹H-NMR spectroscopy.

Electrical and magnetic properties of new compounds of the conductor anion [Ni(dmit)2]- and nitronyl nitroxide magnetic radicals

Three compounds have been synthesized with formulae [3-MeRad][Ni(dmit)2] (1), [4-MeRad][Ni(dmit)2] (2) and [4-PrRad][Ni(dmit)2] (3) where [Ni(dmit)2]- is an anionic pi-radical (dmit = 1,3-dithiol-2-thione-4,5-dithiolate) and [3-MeRad]+ is 3-N-methylpyridinium alpha-nitronyl nitroxide, [4-MeRad]+ is 4-N-methylpyridinium alpha-nitronyl nitroxide and [4-PrRad]+ is 4-N-propylpyridinium alpha-nitronyl nitroxide. The temperature-dependent magnetic susceptibility of 1 revealed that an antiferromagnetic interaction operates between the 3-MeRad+ radical cations with exchange coupling constants of J1 = - 1.72 cm-1 and antiferromagnetism assigned to the spin ladder chains of the Ni(dmit)2 radical anions. Compound 1 exhibits semiconducting behavior and 3 presents capacitor behavior in the temperature range studied (4 - 300 K).

Development of new dissolution test and HPLC-RP method for anti-parasitic ornidazole coated tablets

This work aimed the development and validation of a new dissolution test for ornidazole coated tablets. The dissolution conditions were determined after testing Sink conditions, dissolution medium, apparatus, stirring speed, 24 h stability and medium filtration influence. The best conditions were paddle at a stirring speed of 75 rpm and 900 mL of 0.1 M HCl. A new HPLC quantification method was developed and validated. The dissolution test and quantification method showed to be adequate for their purposes and could be applied for quality control of ornidazole coated tablets, since there is no official monograph.