Lufaxin, a Novel Factor Xa Inhibitor From the Salivary Gland of the Sand Fly Lutzomyia longipalpis Blocks Protease-Activated Receptor 2 Activation and Inhibits Inflammation and Thrombosis In Vivo

Objective-Blood-sucking arthropods' salivary glands contain a remarkable diversity of antihemostatics. The aim of the present study was to identify the unique salivary anticoagulant of the sand fly Lutzomyia longipalpis, which remained elusive for decades. Methods and Results-Several L. longipalpis salivary proteins were expressed in human embryonic kidney 293 cells and screened for inhibition of blood coagulation. A novel 32.4-kDa molecule, named Lufaxin, was identified as a slow, tight, noncompetitive, and reversible inhibitor of factor Xa (FXa). Notably, Lufaxin's primary sequence does not share similarity to any physiological or salivary inhibitors of coagulation reported to date. Lufaxin is specific for FXa and does not interact with FX, Dansyl-Glu-Gly-Arg-FXa, or 15 other enzymes. In addition, Lufaxin blocks prothrombinase and increases both prothrombin time and activated partial thromboplastin time. Surface plasmon resonance experiments revealed that FXa binds Lufaxin with an equilibrium constant approximate to 3 nM, and isothermal titration calorimetry determined a stoichiometry of 1:1. Lufaxin also prevents protease-activated receptor 2 activation by FXa in the MDA-MB-231 cell line and abrogates edema formation triggered by injection of FXa in the paw of mice. Moreover, Lufaxin prevents FeCl3-induced carotid artery thrombus formation and prolongs activated partial thromboplastin time ex vivo, implying that it works as an anticoagulant in vivo. Finally, salivary gland of sand flies was found to inhibit FXa and to interact with the enzyme. Conclusion-Lufaxin belongs to a novel family of slow-tight FXa inhibitors, which display antithrombotic and anti-inflammatory activities. It is a useful tool to understand FXa structural features and its role in prohemostatic and proinflammatory events. (Arterioscler Thromb Vasc Biol. 2012;32:2185-2196.)

A new species of spider fly in the genus Sabroskya Schlinger from Malawi, with a key to Acrocerinae world genera (Diptera, Acroceridae)

In this paper we diagnose the genus Sabroskya Schlinger, 1960 and describe Sabroskya schlingeri sp. n. from Malawi. We also provide dichotomous keys to species of Sabroskya and to world genera of the subfamily Acrocerinae, both extant and extinct.

Is cardiac tissue more susceptible than lung to oxidative effects induced by chronic nasotropic instillation of residual oil fly ash (ROFA)?

The current study aimed to determine the role of oxidants in cardiac and pulmonary toxicities induced by chronic exposure to ROFA. Eighty Wistar rats were divided into four groups: G1 (10 mu L Saline), G2 (ROFA 50 mu g/10 mu L), G3 (ROFA 250 mu g/10 mu L) and G4 (ROFA 500 mu g/10 mu L). Rats received ROFA by nasotropic instillation for 90 days. After that, they were euthanized and bronchoalveolar lavage (BAL) was performed for total count of leukocytes, protein and lactate dehydrogenase (LDH) determinations. Lungs and heart were removed to measure lipid peroxidation (MDA), catalase (CAT) and superoxide dismutase (SOD) activity. BAL presented an increase in leukocytes count in G4 in comparison to the Saline group (p = 0.019). In lung, MDA level was not modified by ROFA, while CAT was higher in G4 when compared to all other groups (p = 0.013). In heart, G4 presented an increase in MDA (p = 0.016) and CAT (p = 0.027) levels in comparison to G1. The present study demonstrated cardiopulmonary oxidative changes after a chronic ROFA exposure. More specifically, the heart tissue seems to be more susceptible to oxidative effects of long-term exposure to ROFA than the lung.

Description of Two New Sand Fly Species Related to Nyssomyia antunesi (Diptera, Psychodidae, Phlebotominae)

Both sexes of two new Brazilian phlebotomine sand flies of the genus Nyssomyia Barretto 1962 [= Lutzomyia (Nyssomyia) sensu Barretto], Nyssomyia delsionatali n. sp. and Nyssomyia urbinattii n. sp., presenting close affinity with Nyssomyia antunesi (Coutinho), are described and illustrated. N. delsionatali n. sp was captured on the edge of a riparian Amazonian forest on the Juruena river in the northwest of Mato Grosso state and N. urbinattii n. sp in a riparian Amazonian forest on the Teles Pires river between Mato Grosso and Para states. Some measurements of both sexes of N. antunesi and illustration of the male genitalia and the female spermathecae as well as an identi_cation key for males and females of the genus Nyssomyia are provided.

INHIBITION OF OVIPOSITION BY NEEM EXTRACT: A BEHAVIORAL PERSPECTIVE FOR THE CONTROL OF THE MEDITERRANEAN FRUIT FLY (DIPTERA: TEPHRITIDAE)

The behavioral inhibitory effect of methanol extracts from neem leaves (Azadirachta indica A. Juss) at different concentrations (0, 10,000, 18,000, 32,000 and 56,000 ppm) was evaluated using naive and experienced medflies (Ceratitis capitata (Wied.)) ovipositing on the fruits of grape cv. 'Italia'. The grapes were immersed in the specific treatments and were exposed for 24 h to 3 pairs of female and male medflies, both nave and experienced, in a choice test. At concentrations >= 18,000 ppm, the extract that was applied to the grapes inhibited oviposition. The previous experience with treated grapes did not affect the response of the medflies. This study is the first step toward the application of the behavior control of the medflies as a tool in grape vineyards. The potential for using oviposition inhibitory in behavioral control of C. capitata are discussed.

Cutting Edge: Brazilian Pemphigus Foliaceus Anti-Desmoglein 1 Autoantibodies Cross-React with Sand Fly Salivary LJM11 Antigen

The environmental factors that contribute to the development of autoimmune diseases are largely unknown. Endemic pemphigus foliaceus in humans, known as Fogo Selvagem (FS) in Brazil, is mediated by pathogenic IgG4 autoantibodies against desmoglein 1 (Dsg1). Clusters of FS overlap with those of leishmaniasis, a disease transmitted by sand fly (Lutzomyia longipalpis) bites. In this study, we show that salivary Ags from the sand fly, and specifically the LJM11 salivary protein, are recognized by FS Abs. Anti-Dsg1 monoclonal autoantibodies derived from FS patients also cross-react with LJM11. Mice immunized with LJM11 generate anti-Dsg1 Abs. Thus, insect bites may deliver salivary Ags that initiate a cross-reactive IgG4 Ab response in genetically susceptible individuals and lead to subsequent FS. Our findings establish a clear relationship between an environmental, noninfectious Ag and the development of potentially pathogenic autoantibodies in an autoimmune disease. The Journal of Immunology, 2012, 189: 1535-1539.

Treatment with probiotics in experimental oral colonization by Candida albicans in murine model (DBA/2)

The aim of this study is to evaluate the oral colonization by Candida albicans in experimental murine immunosuppressed DBA/2 and treatment with probiotic bacteria. To achieve these objectives, 152 DBA/2-immunosuppressed mice were orally inoculated with a suspension of C. albicans containing 10(8) viable yeast cells, the animals were treated with nystatin or with the probiotics (Lactobacillus acidophilus and Lactobacillus rhamnosus). Evaluations were performed by Candida count from oral mucosa swabbing. The oral mucosa colonization by C. albicans started at day 1 after inoculation, remained maximal from day 3 until day 7, and then decreased significantly. Probiotics reduced the C. albicans colonization significantly on the oral mucosa in comparison with the untreated animal group. In the group treated with L. rhamnosus, the reduction in yeast colonization was significantly higher compared with that of the group receiving nystatin. Immunosuppressed animal model DBA/2 is a relevant model for experimental Candida oral colonization, and the treatment with probiotics in this model may be an effective alternative to prevent it. Oral Diseases (2012) 18, 260-264

A remarkable new species of flesh-fly mimicking weevil (Coleoptera: Curculionidae: Conoderinae) from Southeastern Brazil

Timorus sarcophagoides, new species (type-locality: Brazil, Minas Gerais, Santana do Riacho - Serra do Cipo, 43 degrees 35'W 19 degrees 17'S, 1200-1300m ASL), is described and illustrated. The new species can be distinguished mainly from the other species of the genus by the sexual dimorphism of the male rostrum armed with a hooked tubercle at the base of the dorsal carina, while the tubercle is absent in the female and the corresponding region of the carina is only tumid. The new species has a striking pattern of coloration and behavior that mimics flesh-flies in the family Sarcophagidae. Observations on the natural history of the new species are reported and discussed.

Fly ash as zeolites for reducing nitrogen losses by volatilization

The structural and chemical characteristics of fly ash from coal-fired mineral and fly ash zeolitized are similar to those of zeolites. Urea was added with these materials in the proportions of urea: fly ashes of 100:10, 100:20, 100:50, 100:100, with a control containing just urea. These treatments were applied in soil surface and the experimental design was a randomized block with clay and sandy soil. Nitrogen losses by ammonia volatilization and the chemical characteristics of soil fertility were evaluated. In sandy soil there was reduction of ammonia volatilization for the proportions of 100:10 and 100:20, while fly ash zeolitized and fly ash had no difference.

Mass-rearing of Mediterranean fruit fly using low-cost yeast products produced in Brazil

Ceratitis capitata is one of the most important pests of fruits for exportation, and Sterile Insect Technique (SIT) has been the most efficient and environmental friendly technique used to control fruit fly populations around the world. A key goal in achieving a successful SIT program is a mass rearing system producing high quality insects at low cost. Providing adults with an artificial diet containing hydrolysed protein has been the major obstacle for bio-production facilities in Brazil, because it is expensive and has to be imported. Two other commercial products, autolysed yeast (AY) and yeast extract (YE), of domestic origin and low cost, were tested as substitutes of the imported hydrolyzed protein. To compare their efficiency we observed the female fecundity, adult survival and egg viability of flies raised on diets containing one of each of the different protein products. Flies reared on the domestic yeast products had equivalent or superior performance to the flies reared on imported protein. Both AY and YE can be a possible substitute for imported hydrolyzed protein for C. capitata mass-rearing, as they are cheaper and are readily available in the national market.

Colonization of gastropods on subtidal reefs depends on density in adjacent habitats, not on disturbance regime

[EN] Habitats dominated by algal canopies are often altered by physical disturbances of varying severity, changing environmental conditions and biological processes. We used Artificial Seaweed Units (ASUs) to test whether severity of physical disturbances on algal canopies affects the post-disturbance colonization of gastropods on subtidal reefs. Specifically, we examined patterns of assemblage structure of gastropods to test the hypothesis that the extent and intensity of canopy removal affects the post-disturbance colonization of ASUs, testing the consistency of these effects among four regions encompassing a 68 latitudinal gradient in southwestern Australia. Because adjacent habitats can act as a source of new colonists (either as drifting migrants or as a source of propagules) from the perimeter surrounding perturbed areas, we also predicted that patterns of colonization (types and total abundances of colonizers) were influenced by the available pool of individuals at the scale of reefs. Three reefs were selected within each region. On each reef, ASUs were placed in the centre of circular canopy clearings of different size (0, 3, 13 and 50 m2) and intensity (50% vs 100%), and retrieved after 3 months. Resulting assemblages occupying the ASUs were quantitatively representative of the adjacent (undisturbed), algal-associated, assemblages at the scale of reef. Within reefs, recruited assemblages largely mimicked those associated with erect red algae. However, neither disturbance size nor intensity affected the colonization patterns across reefs and regions. These results suggest that algal-associated gastropods, regardless of the prevalent mode of dispersion, are resilient to physical disturbances to canopies across broad geographical regions as long as the pool of potential colonists is maintained. A high dispersal ability of gastropods likely ensures a quick colonization of recovering algal habitats.

Mechanism of colonization in the Mediterranean-sea. Asperarca nodulosa: morphological and genetic analysis

Il mar Mediterraneo è un bacino acquifero peculiare per la recente colonizzazione di specie aliene, per l’evento geologico legato alla Crisi di salinità del Messiniano e per l’ampio range di salinità. L’individuazione dei meccanismi di colonizzazione si è incentrata sullo studio morfologico, istologico e molecolare delle specie Asperarca nodulosa ed Anadara demiri (Arcidae-Bivalvia-Mollusca). La ricerca si è basata sulla caratterizzazione morfologica, con utilizzo del microscopio elettronico a scansione, al fine di individuare il tipo di sviluppo larvale. Successivamente i dati rilevati al S.E.M. sono stati supportati dall’indagine istologica che ha evidenziato la presenza di gonadi a sessi distinti e la non incubazione larvale. L’ulteriore analisi filogenetica ha permesso di evidenziare la netta suddivisione tra le tre popolazioni studiate, indagine effettuata tramite marcatori arbitrari (RAPDs) e nucleari specifici (ITS). I risultati ottenuti trovano supporto da quanto noto su base morfologica. I dati, nel complesso, mostrano una perdita delle capacità di diffusione della specie tramite sviluppo larvale plantotrofico a favore di quello lecitotrofico o diretto; tale tesi è ulteriormente supportata dai dati molecolari che mostrano una netta separazione delle popolazioni prese in esame ed un conseguente isolamento tra individui appartenenti a zone di profondità del Mediterraneo (sub-bacini abissali). La ricerca ha, inoltre,esaminato i meccanismi di introduzione attuali nel bacino acquifero che è soggetto ad una nuova invasione da parte specie aliene dovuta all’apertura del canale di Suez. L’analisi si è focalizzata sullo studio per l’ individuazione dell’origine della specie aliena A. demiri , di presunta derivazione Indo-Pacifica, ma rivelatasi, nei dati preliminari, di origine Atlantica.