Polyunsaturated fatty acids from Phyllocaulis boraceiensis mucus block the replication of influenza virus.

Influenza viruses cause worldwide outbreaks and pandemics in humans and animals every year with considerable morbidity and mortality. The molecular diversity of secondary metabolites extracted from mollusks is a good alternative for the discovery of novel bioactive compounds with unique structures and diverse biological activities. Phyllocaulis boraceiensis is a hermaphroditic slug that exudes mucus, in which was detected hydroxy polyunsaturated fatty acids that exhibited potent antiviral activity against measles virus. The objective of this study was to evaluate this property against Influenza viruses. Cell viability and toxicity of the mucus were evaluated on Madin-Darby canine kidney (MDCK) cells by MTT assay. Antiviral activity from mucus against influenza viruses was carried out by determination of the virus infection dose and by immunofluorescence assays. The crude mucus and its fractions exhibited low cytotoxicity on MDCK cells. A significant inhibition of viral replication, reduced by the order of eight times, was observed in influenza-induced cytopathic effect. In immunofluorescence assay was observed a decrease of more than 80% of the viral load on infected MDCK cell treated with mucus and its fractions. The viral glycoproteins hemagglutinin and neuraminidase located on the surface of the virus are crucial for the replications and infectivity of the influenza virus. Some authors demonstrated that lipids, such as, polyunsaturated fatty acids exhibited multiple roles in antiviral innate and adaptive responses, control of inflammation, and in the development of antiviral therapeutics. As corroborated by other studies, hydroxy polyunsaturated fatty acids interfered with the binding of influenza virus on host cell receptor and reduced viral titers. The results obtained indicated that polyunsaturated fatty acids from P. boraceiensis crude mucus and fractions 39 exerted antiviral activity against influenza virus.

Proteins with Anti-apoptotic Action in the Hemolymph of Caterpillars of the Megalopygidae Family Acts by Maintaining the Structure of the Cellular Cytoskeleton.

Brazil has a very large biological variety, which is an almost inexhaustible source of substances of pharmacological and biotechnological interest. Several studies have demonstrated the presence of bioactive peptides in insect hemolymph and their potential use as therapeutic agents. However, few data are available regarding molecules extracted from insects with anti-apoptotic action. The objective of this work was to identify the presence of proteins from the hemolymph of caterpillars of the Megalopygidae family with pharmacological and biotechnological interest. This study provides preliminary and innovative information on a new substance that inhibits cellular apoptopsis and stabilizes the tested cells, impacting the cytoskeleton, maintaining cellular structure and its functions. To this, two species of Megalopygidae family were studied, Podalia sp. and Megalopyge albicolis. Cytotoxicity tests on Vero and Sf-9 cells revealed that the hemolymph of both caterpillars was cytotoxic only at concentrations greater than 5%v/v. In the anti-apoptotic activity assays, it was verified that the supplementation of cell cultures with only 1% of hemolymph v/v is sufficient to inhibit cell death by apoptosis induced by different inducers such as terbutyl, actinomycin D, hydrogen peroxide, or even by nutrient depletion. For this study, cells were stained with trypan blue, crystal violet, and fluorescent markers to cytoskeleton (actin and tubulin), mitochondria membrane electric potential (JC-1), and apoptosis marker (acridine orange and ethidium). The protein responsible for anti-apoptotic action was isolated through gel filtration chromatography, using an AKTA purifier high-resolution liquid chromatography system. The hemolymph was fractionated into 3 pools for Podalia sp. and 6 pools for M. abicolis. In the antiapoptotic tests, semi-purified hemolymph from both caterpillars showed anti-apoptotic effect in VERO and SF-9 cells, pre-treated with only 1% v/v of hemolymph and induced to death by different and apoptotic inductors. Was observed that the molecule with anti-apoptotic effect is present in pool 3 in both hemolymphs. This protector effect blocked and attenuated the disruption of the cytoskeleton (actin filaments), being that the protective effect also was observed on the integrity of the mitochondrial membrane of SF-9 cells pre-treated with both hemolymphs and treated with the apoptosis inducer Terbutil at concentrations of 25 to 100 µM. By acting on the mitochondrial pathway of death by apoptosis, and by maintaining the structure of the cytoskeleton and cellular functions, pathway that can cause disorders and diseases neurodegenerative, the substances present in the hemolymph of these and other caterpillars could be good candidates in studies for the treatment of neurodegenerative diseases such as Parkinson's disease and Alzheimer's.

Effects of carbohydrate supplementation on competitive runners undergoing overload training followed by a session of intermittent exercise.

This study evaluated the effects of a micro cycle of overload training (1st-8th day) on metabolic and hormonal responses in male runners with or without carbohydrate supplementation and investigated the cumulative effects of this period on a session of intermittent high-intensity running and maximum-performance-test (9th day). The participants were 24 male runners divided into two groups, receiving 61% of their energy intake as CHO (carbohydrate-group) and 54% in the control-group (CON). The testosterone was higher for the CHO than the CON group after the overload training (694.0 +/- 54.6 vs. CON 610.8 +/- 47.9 pmol/l). On the ninth day participants performed 10 x 800 m at mean 3 km velocity. An all-out 1000 m running was performed before and after the 10 x 800 m. Before, during, and after this protocol, the runners received solution containing CHO or the CON equivalent. The performance on 800 m series did not differ in either group between the first and last series of 800 m, but for the all-out 1000 m test the performance decrement was lower for CHO group (5.3 +/- 1.0 vs. 10.6 +/- 1.3%). The cortisol concentrations were lower in the CHO group in relation to CON group (22.4 +/- 0.9 vs. 27.6 +/- 1.4 pmol/l) and the IGF1/IGFBP3 ratio increased 12.7% in the CHO group. During recovery, blood glucose concentrations remained higher in the CHO group in comparison with the CON group. It was concluded that CHO supplementation possibly attenuated the suppression of the hypothalamic-pituitary-gonadal axis and resulted in less catabolic stress, and thus improved running performance.

Patagonin-CRISP: Antimicrobial Activity and Source of Antimicrobial Molecules in Duvernoy's Gland Secretion (Philodryas patagoniensis Snake).

Snake venom contains a variety of toxins with a range of biological activity, among these toxins cysteine-rich secreted proteins (CRISPs) can be found. The proteins of this family have masses of 20-30 kDa and display homologous amino acid sequences containing 16 cysteine residues, forming eight disulfide bonds. Some of these proteins have been explored, characterized, and described in terms of their activity; however, little is known about their range of activities. A search for new antimicrobial molecules is ongoing, as the number of microbial strains resistant to available antibiotics is increasing. We identified antimicrobial activity in the secretion of Duvernoy's gland of the rear-fanged Philodryas patagoniensis. Fractions of this venom were subjected to reverse-phase high performance liquid chromatography and analyzed to determine their antimicrobial activity with a liquid broth inhibition assay. One of the fractions presented activity against a Gram-negative bacterium and a filamentous fungus. This fraction was analyzed with LC-MS/MS, and a protein of 24,848.8 Da was identified. Database searches allowed us to identify it as a CRISP due to the presence of some unique fragments in the molecule. We called it patagonin-CRISP, as the same protein in the venom of P. patagoniensis had previously been characterized as having a different biological activity. Patagonin-CRISP presented activity at very low concentrations and showed no cytotoxic activity. This is the first time that antimicrobial activity has been identified for P. patagoniensis venom or for a CRISP family protein.

Evaluación del efecto inmunomodulador de Bougainvillea xbuttiana (var. naranja) sobre la producción de citoquinas de macrófagos inducida pelo veneno de Bothrops jararaca

To study the effect of 50% ethanol extract of Bougainvillea xbuttianaon the enzymatic activity, cell viability and cytokine production provoked by the venom of Bothrops jararacain macro-phages. Three assays were used to study the effects of B. xbuttianaextract on the damage pro-duced by B. jararaca: Enzymatic activity was detected by measuring the proteolytic and phos-pholipase A2; macrophages cytotoxicity was determined by the MTT method; levels of cytokine were evaluated using ELISA and a biological assay. After treatment with 300 μg/mL B. xbuttianaextract for 30 min, the proteolytic and phospholipase A2activities of the venom were reduced to 95 and 61%, respectively. In macrophages cultures treated with B. xbuttianaextract combined with venom, the production of TNF-α, IL-6 and IFN-γ was reduced, whereas IL-10 was potenti-ated. Our results support the potential effect of the B. xbuttianaextract as a complementary therapy against the toxicity caused by the venom of B. jararacasnakes.

Antiviral action of aqueous extracts of propolis from Scaptotrigona aff. postica (Hymenoptera; Apidae) against Zica, Chikungunya, and Mayaro virus

The limited availability of antivirals for new highly pathogenic strains of virus has become a serious public health. Therefore, news products against these pathogens has become an urgent necessity. Among the multiple sources for news antibiotics and antivirals, insect exudates or their products has become an increasingly frequent option. Insects emerged 350 million years ago and have showed a high adaptability and resistance to the most varied biomes. Their survival for so long, in such diferent environments, is an indication that they have a very efcient protection against environmental infections, despite not having a developed immune system like mammals. Since the ancient civilizations, the products obtained from the bee have been of great pharmacological importance, being used as antimicrobial, anti-infammatory, antitumor and several other functions. Investigations of biological activity of propolis have been carried out, mainly in the species Apis mellifera, and its product have showed activity against some important viruses. However, for the Meliponini species, known as stingless bees, there are few studies, either on their chemical composition or on their biological activities. The importance of studying these bees is because they come from regions with native forests, and therefore with many species of plants not yet studied, in addition to which they are regions still free of pesticides, which guarantees a greater fdelity of the obtained data. Previous studies by our group with crude hydroalcoholic extract of propolis demonstrated an intense antiviral activity against Herpes, infuenza, and rubella viruses. In this work, we chose to use aqueous extracts, which eliminates the presence of other compounds besides those originally present in propolis, in addition to extracting substances diferent from those obtained in alcoholic extracts. Therefore, this study aimed to identify, isolate and characterize compounds with antiviral efects from aqueous propolis extracts from Scaptotrigona af postica, in emerging viruses such as zicavirus, chikungunya, and mayaro virus. The evaluation of the antiviral activity of the crude and purifed material was performed by reducing infectious foci in VERO cell cultures. The results obtained with crude propolis, indicate a high reduction of zica virus (64×) and mayaro (128×) when was used 10% v/v of propolis. The reduction of chikungunya virus was of 256 fold, even when was used 5% v/v of propolis. The chemical characterization of the compounds present in the extracts was performed by high-pressure liquid chromatography. Through the purifcation of propolis by HPLC and mass spectrometry, it was possible to identify and isolate a peak with antiviral activity. This substance showed activity against all viruses tested. When purifed fraction was used, the reduction observed was of 16 fold for zicavirus, 32 fold for mayaro virus and 512 fold for chikungunya virus. Likewise, it was observed that the antiviral response was concentration dependent, being more intense when propolis was added 2 h after the viral infection. Now we are carrying out the chemical characterization of the purifed compounds that showed antiviral action.

Antiproliferative and antiangiogenic effect of Amblyomma sculptum (Acari: Ixodidae) crude saliva in endothelial cells in vitro.

Neovascularization, a process that includes vasculogenesis and angiogenesis, may be a physiological or pathologic event, but in any cases the phenomenon is related to the formation of vascular net and sprouting of endothelial cells from preexisting blood vessel. The tumor environment, which counts on the tumor cell proliferation, is plenty of proangiogenic factors, such as angiogenin, TGF (α and ß), FGF, VEGF, all of them playing a crucial role in angiogenesis, an important hallmark of cancer frequently related to a poor prognosis. Therefore, therapies focusing the inhibition of cancer neovasculogenesis have become an interesting strategy for the development of antitumor therapies. In this work, we investigate the effect of tick saliva on the human endothelial cells, in order to understand its inhibitory effects on angiogenesis. To this end, the HUVEC cells were used as model of angiogenesis in vitro and the anti-proliferative, anti-migratory, cytotoxicity was evaluated. Our data depicts that saliva impairs cell development by causing structural changes while precludes cell proliferation and migration, that are crucial events related to angiogenesis. Aiming the identification of the bioactive components related to antiangiogenic activity, saliva was analyzed through the Mass Spectrometry and among all molecules identified, disintegrins and cathepsin L seems to be primarily responsible for the antiangiogenic effects of saliva.

Proteins with anti-apoptotic action in the hemolymph of caterpillars of the Megalopygidae family acts by maintaining the structure of the cellular cytoskeleton

Brazil has a very large biological variety, which is an almost inexhaustible source of substances of pharmacological and biotechnological interest. Several studies have demonstrated the presence of bioactive peptides in insect hemolymph and their potential use as therapeutic agents. However, few data are available regarding molecules extracted from insects with anti-apoptotic action. The objective of this work was to identify the presence of proteins from the hemolymph of caterpillars of the Megalopygidae family with pharmacological and biotechnological interest. This study provides preliminary and innovative information on a new substance that inhibits cellular apoptopsis and stabilizes the tested cells, impacting the cytoskeleton, maintaining cellular structure and its functions. To this, two species of Megalopygidae family were studied, Podalia sp. and Megalopyge albicolis. Cytotoxicity tests on Vero and Sf-9 cells revealed that the hemolymph of both caterpillars was cytotoxic only at concentrations greater than 5%v/v. In the anti-apoptotic activity assays, it was verified that the supplementation of cell cultures with only 1% of hemolymph v/v is sufficient to inhibit cell death by apoptosis induced by different inducers such as terbutyl, actinomycin D, hydrogen peroxide, or even by nutrient depletion. For this study, cells were stained with trypan blue, crystal violet, and fluorescent markers to cytoskeleton (actin and tubulin), mitochondria membrane electric potential (JC-1), and apoptosis marker (acridine orange and ethidium). The protein responsible for anti-apoptotic action was isolated through gel filtration chromatography, using an AKTA purifier high-resolution liquid chromatography system. The hemolymph was fractionated into 3 pools for Podalia sp. and 6 pools for M. abicolis. In the antiapoptotic tests, semi-purified hemolymph from both caterpillars showed anti-apoptotic effect in VERO and SF-9 cells, pre-treated with only 1% v/v of hemolymph and induced to death by different and apoptotic inductors. Was observed that the molecule with anti-apoptotic effect is present in pool 3 in both hemolymphs. This protector effect blocked and attenuated the disruption of the cytoskeleton (actin filaments), being that the protective effect also was observed on the integrity of the mitochondrial membrane of SF-9 cells pre-treated with both hemolymphs and treated with the apoptosis inducer Terbutil at concentrations of 25 to 100 µM. By acting on the mitochondrial pathway of death by apoptosis, and by maintaining the structure of the cytoskeleton and cellular functions, pathway that can cause disorders and diseases neurodegenerative, the substances present in the hemolymph of these and other caterpillars could be good candidates in studies for the treatment of neurodegenerative diseases such as Parkinson’s disease and Alzheimer’s.

Antiviral activity of red propolis against herpes simplex virus-1

Brazilian Red Propolis is a resinous material produced by Apis mellifera bees through the collection of the exudates of Dalbergia ecastaphyllum, rich in flavonoids, and Symphonia globulifera and Clusia species, which are rich in polyprenylated benzophenones. More than 200 compounds, including flavonoids and polyprenylated benzophenones have been found in Brazilian red propolis. The objective of the present study was to evaluate the chemical composition and antiviral activity of an ethanolic extract of red propolis from Alagoas, Brazil, against Herpes Simplex Virus (HSV-1). In the HPLC-PDA-ESI-MS/MS analysis were detected flavanones, isoflavones, chalcones, pterocarpans and polyprenylated benzophenones. The measurement of antiviral activity of red propolis extract was performed by DNA quantification through quantitative real-time PCR assay and negative staining Electron Microscopy. The pretreatment, post-treatment, and virucide assays using ethanolic extract of red propolis with concentration of 8, 12, 24, 48, or 96 µg/mL, indicated an inhibition of the viral binding and viral entry into cells as well as the replication of HSV-1. In the electron microscopy imaging was observed the disruption of the viral membrane in the HSV-1 treated with red propolis, when compared with HSV-1 that was treated with phosphate buffered saline alone, indicating that ethanolic extract from red propolis can act directly on the viral envelope, through lipid membrane degradation and/or directly blocking the enriched proteins on the viral surface.

Antitumoral effects of Amblyomma sculptum Berlese saliva in neuroblastoma cell lines involve cytoskeletal deconstruction and cell cycle arrest.

The antitumor properties of ticks salivary gland extracts or recombinant proteins have been reported recently, but little is known about the antitumor properties of the secreted components of saliva. The goal of this study was to investigate the in vitro effect of the saliva of the hard tick Amblyomma sculptum on neuroblastoma cell lines. SK-N-SK, SH-SY5Y, Be(2)-M17, IMR-32, and CHLA-20 cells were susceptible to saliva, with 80% reduction in their viability compared to untreated controls, as demonstrated by the methylene blue assay. Further investigation using CHLA-20 revealed apoptosis, with approximately 30% of annexin-V positive cells, and G0/G1-phase accumulation (>60%) after treatment with saliva. Mitochondrial membrane potential (Δψm) was slightly, but significantly (p < 0.05), reduced and the actin cytoskeleton was disarranged, as indicated by fluorescent microscopy. The viability of human fibroblast (HFF-1 cells) used as a non-tumoral control decreased by approximately 40%. However, no alterations in cell cycle progression, morphology, and Δψm were observed in these cells. The present work provides new perspectives for the characterization of the molecules present in saliva and their antitumor properties.

Antimicrobial activity of flavonoids glycosides and pyrrolizidine alkaloids from propolis of Scaptotrigona aff. postica

Stingless bees, or simply meliponines, are a large group of bees (about 550 described species), comprising the tribe Meliponini. They belong in the family Apidae, and are closely related to Apis mellifera. Scaptotrigona affinis postica Latreille, 1807 from northeast of Brazil is popularly known as “tubi” in Maranhão State. Flavones di-C-glycosides, and the pyrrolizidine alkaloid 3-methoxy-7-(2-methylbutyryl)-9-echimidinyl retronecine (7) were reported previously in propolis from S. postica and detected only in samples collected in October, November and December. The presence of pyrrolizidine alkaloids and their N-oxides were reported in honey and pollen. In the present study, the preparative liquid chromatography in reversed-phase column from ethanolic extract of propolis allowed the separation of fractions 40AEP and 40 MEP, which were analyzed by LC-MS. Many pyrrolizidine alkaloids, among them, lithosenine (14), lithosenine arabinoside (19), 7-angeloyl-9-(2,3- dihydroxybutyryl) retronecine (1), 7-(2- methylbutyryl) retronecine (3), 9-sarracinoyl retronecine (13) and viridinatine (8), besides the flavonoids schaftoside (15), aromadendrin-7-O-methyl ether (12), 7- methoxy-5,6,3’,4’,5’,6’-hexahydroxy-flavone-3-O-glucuronide (11), mangiferin (10) and mangiferin-O-methyl ether (17) were detected in these fractions. Fraction 40MEP showed antibacterial effect against gram positive and gram negative bacterial pathogens with MIC ranging from 62.5 μg/mL to 200 μg/mL, while fractions 40AEP and 40MEP showed antimicrobial activity against gram negative bacteria, including Escherichia coli D31- streptomycin resistant. The cells were treated with 40MEP and 40AEP at a concentration of 200 μg/mL and were not observed statistical difference between treated and untreated cells.

A highly complex stingless bee propolis: composition and influence of the period of collection

BACKGROUND Stingless bees or meliponines (family Apidae, subfamily Meliponinae, tribe Meliponini) are eusocial bees from tropical and subtropical regions. Propolis of Scaptotrigona aff. Postica (Latreille, 1807) is used in the state of Maranhão (Northeast Brazil) in ointments to treat tumors and wounds. Samples of propolis of Scaptotrigona aff. Postica (Apidae, Meliponini) were collected monthly from an apiary located in Barra do Corda (state of Maranhão, northeast Brazil). Extracts of the twelve samples were obtained with 80% ethanol. Constituents of the samples were characterized by HPLC-PAD-ESI-MS/MS analysis, amounting to 100 substances. RESULTS Representatives of several classes of secondary metabolites were characterized, including benzoic and cinnamic acids, flavonoids (chalcones, flavone-C-glycosides, flavonol aglycones and glycosides), alkyl and alkenyl resorcinols, xanthones, diterpenes, cycloartane-type triterpenoids, pentacyclic triterpenoids, pyrrolizidine alkaloids and hydroxycinnamic acid amides (HCAAs). Considerable qualitative differences in chemical composition among samples were observed, depending on the year period of collection. Principal Coordinate Analysis recognized three distinct year periods (Jan-Mar, April-Sep, Oct-Dec) according to the corresponding chemical profiles. CONCLUSION Comparing with previous studies, the present work indicates that considerable differences in chemical composition occur also from year to year. Contrary to most propolis types reported so far, which derives exclusively or mostly from a single botanical source, the propolis from Barra do Corda seemingly depends on several resin sources. It is suggested that chalcones and flavonols stem from Mimosa tenuiflora (Mimosoideae); resorcinols, xanthones and cycloartane-type triterpenoids, from fruits of Mangifera indica (Anacardiaceae); pyrrolizidine alkaloids, possibly from some Crotalaria species (Faboideae); HCAAs probably originate from pollen contaminating the propolis samples. The propolis of S. aff. Postica poses challenges and possibilities of study for apicultural researchers, chemists and pharmacologists.

Gut transcriptome analysis on females of Ornithodoros mimon (Acari: Argasidae) and phylogenetic inference of ticks.

Ornithodoros mimon is an argasid tick that parasitizes bats, birds and opossums and is also harmful to humans. Knowledge of the transcripts present in the tick gut helps in understanding the role of vital molecules in the digestion process and parasite-host relationship, while also providing information about the evolution of arthropod hematophagy. Thus, the present study aimed to know and ascertain the main molecules expressed in the gut of argasid after their blood meal, through analysis on the gut transcriptome of engorged females of O. mimon using 454-based RNA sequencing. The gut transcriptome analysis reveals several transcripts associated with hemoglobin digestion, such as serine, cysteine, aspartic proteases and metalloenzymes. The phylogenetic analysis on the peptidases confirmed that most of them are clustered with other tick genes. We recorded the presence a cathepsin O peptidase-coding transcript in ticks. The topology of the phylogenetic inferences, based on transcripts of inferred families of homologues, was similar to that of previous reports based on mitochondrial genome and nuclear rRNA sequences. We deposited 2,213 sequence of O. mimon to the public databases. Our findings may help towards better understanding of important argasid metabolic processes, such as digestion, nutrition and immunity.

Evaluation of genotoxic and cytogenetic effects of saponins and aluminum hydroxide adjuvant on viral vaccine

Bovine papillomavirus (BPV) is the etiological agent of bovine papillomatosis, infectious disease characterized by the presence of benign tumors that can progress to malignancy. The phylogenetic classification of the PVs is performed based on the sequence homology of the Open Reading Frame L1, the most conserved among different viral serotypes. Given the immunogenicity of saponins, it,s has been used as a candidate as adjuvant use. For this reason, the safety of using saponin as an adjuvant has to be better determined to human or veterinary vaccine use. So, this study aimed to evaluate the mutagenic and genotoxic effect of saponins in comparison with the adjuvant widely used aluminum hydroxide using an isolated and purified L1 protein from BPV as model. In this study, genomic lesions, which after processed without repair can result in mutations, were detected by comet assay. Possible damages to genetic material caused by structural chromosomal changes (clastogenesis), as well as chromosomal losses (aneugenesis) were evaluated by the micronucleus test. Both tests were done on polychromatic erythrocytes and Vero cells. The evaluation of apoptosis and necrosis of treated Vero cells was made by Annexin V / PI staining and flow cytometry. The two vaccine products (L1 + Saponin and L1 + Aluminum Hydroxide) showed damages compatible with the positive control in the comet assay and both slightly elevated the micronucleus levels, in the Cell Viability Assay the results with Aluminum Hydroxide were satisfactory, characterizing Aluminum Hydroxide as a safer adjuvant according to the proposed tests, better than the saponins. Some fractions of the saponin extract separated by High Performance Liquid Chromatography were evaluated against genotoxic activity by comet assay, and their identities were confirmed by similarity to the reference standard by mass spectrometry.

Historical analysis of leishmaniasis cases in the transamazonian region: from 2009 to 2019 / Análise histórica dos casos de leishmaniose na região transamazônica: de 2009 a 2019 / Análisis histórico de casos de leishmaniasis en la región transamazónica: de 2009 a 2019

Objective: To determine the total number of notified cases, new cases and recurrence of American Tegumentary Leishmaniasis (ATL), to investigate the association between HIV coinfection and the presence of cutaneous lesion, as well to assess the concordance of two different laboratorial procedures: parasitological and histopathological in this region. Methods:This is a cross-sectional study. The research used data from the Information System for Notifiable Diseases, hence forth designated as SINAN (Sistema de Informação de Agravos de Notificação) that contained 6,183 cases of leishmaniasis in 9 municipalities. Results:As results, 5795 were of new cases and 351 recurrence cases. Among the cases 1,146 patients were tested for HIV, 16 presented coinfection from these 15 developed cutaneous lesion. In the overall studied population 5,690 subjects manifested cutaneous lesion. There was no agreement of the results regarding the quality of the parasitological and histopathological exams, demonstrating that they are poorly replicable (p <0.005). Conclusion:The ATL is endemic in the transamazonian region posing as a major public health problem. All patients with leishmaniasis must be tested for HIV and health professionals must register on the notification form. The tests to diagnose leishmaniasis need to be more specific and sensitive.

Objetivo: Determinar o número total de casos notificados, casos novos e recorrência da Leishmaniose Tegumentar Americana (LTA), investigar a associação entre a coinfecção pelo HIV e a presença de lesão cutânea, bem como avaliar a concordância de dois procedimentos laboratoriais distintos: parasitológico e histopatológico nessa região. Métodos: Este é um estudo transversal. A pesquisa utilizou dados do Sistema de Informação de Agravos de Notificação, doravante denominado SINAN, que continha 6.183 casos de leishmaniose em 9 municípios. Resultados: Como resultados, 5795 eram de casos novos e 351 casos de recorrência. Dos casos 1.146 pacientes testados para HIV, 16 apresentaram coinfecção, destes 15 desenvolveram lesão cutânea. Na população geral estudada 5.690 indivíduos manifestaram lesão cutânea. Não houve concordância dos resultados quanto à qualidade dos exames parasitológicos e histopatológicos, demonstrando que são pouco replicáveis ​​(p <0,005). Conclusão: A ATL é endêmica na região transamazônica e constitui um importante problema de saúde pública. Todos os pacientes com leishmaniose devem ser testados para HIV e os profissionais de saúde devem se cadastrar na ficha de notificação. Os testes para diagnosticar a leishmaniose precisam ser mais específicos e sensíveis.

Rondonin: antimicrobial properties and mechanism of action

Infectious diseases are among the major causes of death in the human population. A wide variety of organisms produce antimicrobial peptides (AMPs) as part of their first line of defense. A peptide from Acanthoscurria rondoniae plasma, rondonin – with antifungal activity, a molecular mass of 1,236 Da and primary sequence IIIQYEGHKH – was previously studied (Uniprot accession number B3EWP8). It showed identity with the C-terminus of subunit "D" of the hemocyanin of the Aphonopelma hentzi spider. This result led us to propose a new pathway of the immune system of arachnids that suggests a new function to hemocyanin: production of antimicrobial peptides. Rondonin does not interact with model membranes and was able to bind to yeast nucleic acids but not bacteria. It was not cytotoxic against mammalian cells. The antifungal activity of rondonin is pH dependent and peaks at pH ~4-5. The peptide presents synergism with Gomesin (spider hemocyte antimicrobial peptide - UniProtKB - P82358) against human yeast pathogens, suggesting a new potential alternative treatment option. Antiviral activity was detected against RNA viruses, measles, H1N1 and encephalomyocarditis. This is the first report of an arthropod hemocyanin fragment with activity against human viruses. Currently, it is vital to invest in the search for natural and synthetic antimicrobial compounds that, above all, present alternative mechanisms of action to first-choice antimicrobials.

Patagonin-CRISP: antimicrobial activity and source of antimicrobial molecules in duvernoy's gland secretion (Philodryas patagoniensis Snake)

Snake venom contains a variety of toxins with a range of biological activity, among these toxins cysteine-rich secreted proteins (CRISPs) can be found. The proteins of this family have masses of 20–30 kDa and display homologous amino acid sequences containing 16 cysteine residues, forming eight disulfide bonds. Some of these proteins have been explored, characterized, and described in terms of their activity; however, little is known about their range of activities. A search for new antimicrobial molecules is ongoing, as the number of microbial strains resistant to available antibiotics is increasing. We identified antimicrobial activity in the secretion of Duvernoy's gland of the rear-fanged Philodryas patagoniensis. Fractions of this venom were subjected to reverse-phase high performance liquid chromatography and analyzed to determine their antimicrobial activity with a liquid broth inhibition assay. One of the fractions presented activity against a Gram-negative bacterium and a filamentous fungus. This fraction was analyzed with LC-MS/MS, and a protein of 24,848.8 Da was identified. Database searches allowed us to identify it as a CRISP due to the presence of some unique fragments in the molecule. We called it patagonin-CRISP, as the same protein in the venom of P. patagoniensis had previously been characterized as having a different biological activity. Patagonin-CRISP presented activity at very low concentrations and showed no cytotoxic activity. This is the first time that antimicrobial activity has been identified for P. patagoniensis venom or for a CRISP family protein.

The antiviral effect of mollusk mucus on measles virus.

Measles is a viral disease highly contagious spread by respiratory transmission. Although infection can be controlled by vaccination, numerous cases of measles have been registered in many areas of the world, highlighting the need for additional interventions. Terrestrial gastropods exude mucus on their body surface when traveling, to protect the body from mechanical injury, desiccation or contact with harmful substances. The mucus of mollusks has been studied as a source of new natural compounds with diverse biological activities. In this study, the antiviral activity of the mucus of the land slug P. boraceiensis was demonstrated in vitro using Vero cells infected with measles virus. The crude sample and four fractions were tested in cultures infected with measles virus and the antiviral activity was assessed by the cytopathic effect in infected cell cultures as well as by immunofluorescence and qPCR. Fractions 39 and 50 of the mucus from P. boraceiensis were analyzed by HPLC-DAD-ESI-MS/MS and infrared spectroscopy. A mixture of polyunsaturated fatty acids was found in the two fractions. A reduction in the growth of the measles virus was observed, measured by qPCR, with a protection index of 80% in Vero cells infected with measles and treated with fraction 39. Fraction 39 exhibited the best antiviral action in vitro and high contents of hydroxy-tritriacontapentaenoic acid and hydroxy-pentatriacontapentaenoic acid were found in this fraction.

Co-infection between influenza virus and flagellated bacteria.

Trypsin is required in the hemagglutinin (HA) cleavage to in vitro influenza viruses activation. This HA cleavage is necessary for virus cell entry by receptor-mediated endocytosis. Bacteria in the respiratory tract are potential sources of proteases that could contribute to the cleavage of influenza virus in vivo. From 47 samples collected from horses, pigs, and from humans, influenza presence was confirmed in 13 and these samples demonstrated co-infection of influenza with flagellated bacteria, Stenotrophomonas maltophilia from the beginning of the experiments. Despite treatment with antibiotics, the bacteria remained resistant in several of the co-infected samples (48.39%). These bacteria, considered opportunistic invaders from environmental sources, are associated with viral infections in upper respiratory tract of hosts. The protease (elastase), secreted by Stenotrophomonas maltophilia plays a role in the potentiation of influenza virus infection. Proteolytic activity was detected by casein agar test. Positive samples from animals and humans had either a potentiated influenza infectivity or cytopathic effect (CPE) in MDCK and NCI H292 cells, Stenotrophomonas maltophilia were always present. Virus and bacteria were observed ultrastructurally. These in vitro findings show that microbial proteases could contribute to respiratory complications by host protease activity increasing inflammation or destroying endogenous cell protease inhibitors.

Anti-inflammatory effect of Bougainvillea × buttiana (Var. Orange) extract in experimental Bothrops jararaca envenomation

Background: Bougainvillea x buttiana is a plant used in folk Mexican medicine to treat different inflammatory diseases. Objective: In this study, the anti-inflammatory effect of B. x buttiana orange extract (BxbO) was evaluated on edema formation, cytokine production, and lethality in mice in response to venom of Bothrops jararaca (VBj) snake. Materials and Methods: The BxbO extract was tested in vitro to determine its effect on phospholipase A2(PLA2) and in vivo for the formation of edema, the paw edema model was used, as well as the toxicity of the extract and the production of cytokines. Lethality induced by VBj, the survival percentage, was calculated. Results: BxbO extract significantly inhibited in vitro PLA2and in vivo, blocked the edema formation and cytokine production, and prevented lethality induced by VBj. The constituents of BxbO may bind to components of VBj, such as PLA2, thereby blocking the proteolytic action of the venom. In animals treated with BxbO extract injected 1 h after the venom injection, no difference was observed in the cytokine secretion. In contrast, for all mice treated with BxbO extract for 1 h before VBj administration or together with VBj, the pro-inflammatory cytokine secretion in the serum was attenuated and an exacerbated production anti-inflammatory cytokine. In the presence of the BxbO extract injected 1 h before the VBj injection or together with the VBj injection, mortality was significantly lower. Conclusion: Altogether, our results show that BxbO extract can inhibit the local and systemic activities of VBj. However, new studies are still required to identify the interaction mechanisms between bioactive compounds and cellular components.