WorldWideScience

Sample records for pp71 ppul82 tegument

  1. Nuclear trafficking of the human cytomegalovirus pp71 (ppUL82) tegument protein

    International Nuclear Information System (INIS)

    Shen Weiping; Westgard, Elizabeth; Huang Liqun; Ward, Michael D.; Osborn, Jodi L.; Chau, Nha H.; Collins, Lindsay; Marcum, Benjamin; Koach, Margaret A.; Bibbs, Jennifer; Semmes, O. John; Kerry, Julie A.

    2008-01-01

    The human cytomegalovirus tegument protein pp71 localizes to the nucleus immediately upon infection, and functions to initiate viral gene expression. Analysis of a series of random insertion mutations revealed that sequences within the mid region (MR) of pp71 are important for localization to the nucleus. Fusion of MR sequences with eGFP revealed that amino acids 94 to 300 were sufficient to target proteins to the nucleus. Random substitution mutagenesis within this domain resulted in two double substitution mutants, pp71P203T/T223M and pp71T228M/L275Q, with a predominantly cytoplasmic localization. Disruption of nuclear targeting resulted in relocalization of the fusion proteins to a distinct perinuclear region. Using tandem mass spectrometry, we determined that threonine 223 can be phosphorylated. Mutation of this residue to a phosphomimetic amino acid resulted in abrogation of nuclear targeting. These results strongly suggest that the intracellular trafficking of pp71 is regulated by phosphorylation

  2. Analisis Implementasi Penerapan Standar Akuntansi Pemerintah Berbasis Akrual Sesuai dengan Peraturan Pemerintah ( Pp ) 71 Tahun 2010 ( Study pada Pemerintah Daerah Kabupaten Karimun )

    OpenAIRE

    ', Supriyono; ', Desmiyawati; Azlina, Nur

    2016-01-01

    The formulation of the research problem is as follows: 1). How readiness district government that Karimun indicated by the commitment, human resources, infrastructure and information systems in mengimplementa sikan PP 71 Year 2010 About the Government Accounting Standards (SAP) Based Accrual?, What are the obstacles in the implementation of Regulation No. 71 Year 2010, on Accounting Standards Government (SAP)? 2). What is an obstacle in the implementation of Regulation No. 71 Year 2010, conce...

  3. Tegumental histological effects of Mirazid(®) and myrrh volatile oil on adult Fasciola gigantica.

    Science.gov (United States)

    Massoud, Ahmad Mohamed; Shalaby, Hatem Abdel Mawgoud; El Khateeb, Rabab Mohamed; Mahmoud, Mona Said; Kutkat, Mohamed Abdel Aziz

    2013-06-01

    To evaluated the histological changes within the tegument of adult Fasciola gigantica (F. gigantica) that led to the gross changes that were visible externally. The effects of oleoresin extract of myrrh (Mirazid(®)), myrrh volatile oil and triclabendazole sulphoxide (reference drug) on the tegumental structure of adult F. gigantica following treatment in vitro had been determined by light microscopy. The internal changes in the tegument observed in this study were compatible with surface changes seen in the previous scanning electron microscopy study, using the same drugs. The swelling of tegumental syncytium was a particular feature of their action, but its level was much greater with myrrh volatile oil, in which vacuolization of the tegument and loss of spines were observed. The present study demonstrated the fasciocidal properties of Mirazid(®) oleoresin extract, and it might be possible to reinforce its fasciocidal activity by increasing its content of myrrh volatile oil.

  4. Surface histology, topography, and ultrastructure of the tegument of adult Orthocoelium parvipapillatum (Stiles & Goldberger, 1910).

    Science.gov (United States)

    Anuracpreeda, Panat; Chawengkirttikul, Runglawan; Sobhon, Prasert

    2016-07-01

    Adult Orthocoelium parvipapillatum are common parasites that reside in the rumen and reticulum of ruminants, i.e., cattle, sheep, goats, and buffaloes. The fluke is conical-shaped and slightly concave ventrally and convex dorsally, and measures bout 2.4-3.9 mm in length and 1.0-2.3 mm in width across the mid-section. The tegument of the adult worm is examined using light microscopy (LM) and scanning (SEM) and transmission electron microscopy (TEM). Under LM, the tegument appears as a thick homogeneous layer containing folds alternated with grooves without spines. SEM revealed that the tegumental surface is highly corrugated with ridges and furrows and appears spineless. Two types of sensory papillae are observed, i.e., type 1 is bulbous in shape with nipple-like tips and type 2 has a similar shape with short cilia. In TEM, the tegument has a typical syncytial organization and is divided into four layers. The first layer of the tegument contains ridges and furrows covered by a trilaminate membrane coated externally with the glycocalyx. The second layer is a strait area of cytoplasm that includes numerous ovoid electron-lucent (TG1) and disc-shaped electron-dense (TG2) tegumental granules and lysosomes. The third layer is the widest middle area which contains several evenly distributed mitochondria, TG1 and TG2. The fourth layer rests on a thick basal lamina and contains numerous infoldings of the basal plasma membrane with closely associated mitochondria. Both granules are produced and transported to the tegument by one type of tegumental cells lying in rows below the muscular layers.

  5. Role of a reducing environment in disassembly of the herpesvirus tegument

    Energy Technology Data Exchange (ETDEWEB)

    Newcomb, William W. [Department of Microbiology Immunology and Cancer Biology, University of Virginia Health System, Box 800734, University of Virginia Health System, 1300 Jefferson Park Ave. Charlottesville, VA 22908 (United States); Jones, Lisa M. [Department of Chemistry, Washington University in St. Louis, St. Louis, MO 63130 (United States); Dee, Alexander [Department of Molecular, Microbial and Structural Biology, University of Connecticut Health Center, Farmington, CT 06030 (United States); Chaudhry, Farid [Department of Microbiology Immunology and Cancer Biology, University of Virginia Health System, Box 800734, University of Virginia Health System, 1300 Jefferson Park Ave. Charlottesville, VA 22908 (United States); Brown, Jay C., E-mail: JCB2G@VIRGINIA.EDU [Department of Microbiology Immunology and Cancer Biology, University of Virginia Health System, Box 800734, University of Virginia Health System, 1300 Jefferson Park Ave. Charlottesville, VA 22908 (United States)

    2012-09-15

    Initiation of infection by herpes family viruses involves a step in which most of the virus tegument becomes detached from the capsid. Detachment takes place in the host cell cytosol near the virus entry site and it is followed by dispersal of tegument proteins and disappearance of the tegument as a distinct entity. Here we describe the results of experiments designed to test the idea that the reducing environment of the cytosol may contribute to tegument detachment and disassembly. Non-ionic detergent was used to remove the membrane of purified herpes simplex virus under control and reducing conditions. The effects on the tegument were then examined by SDS-PAGE and electron microscopy. Protein analysis demonstrated that most major tegument proteins were removed under both oxidizing and reducing conditions except for UL49 which required a reducing environment. It is proposed therefore that the reducing conditions in the cytosol are involved in removal of UL49 protein. Electron microscopic analysis revealed that capsids produced under oxidizing conditions contained a coating of protein that was absent in reduced virions and which correlated uniquely with the presence of UL49. This capsid-associated layer is suggested to be the location of UL49 in the extracted virion.

  6. Anti-inflammatory polysaccharides of Azadirachta indica seed tegument

    Directory of Open Access Journals (Sweden)

    Lívia de Paulo Pereira

    2012-06-01

    Full Text Available Azadirachta indica A. Juss., Meliaceae, or Indian neem is a plant used to treat inûammatory disorders. Total polysaccharide (TPL and FI (fractioned by ion exchange chromatography from the seed tegument of A. indica were evaluated in models of acute inflammation (paw edema/peritonitis using Wistar rats. Paw edema (measured by hydroplethysmometry was induced s.c. by Λ-carrageenan (300 µg, histamine (100 µg, serotonin (20 µg, compound 48/80 (10 µg, prostaglandin (PGE2 30 µg or L-arginine (15 µg. Peritonitis (analyzed for leukocyte counts/protein dosage was induced i.p. by carrageenan (500 mg or N-formyl-methionyl-leucyl-phenylalanine (fMLP 50 ng. Animals were treated i.v. with TPL (1 mg/kg or FI (0.01, 0.1, 1 mg/kg 30 min before stimuli. FI toxicity (at 0.1 mg/kg, i.v. for seven days was analyzed by the variation of body/organ mass and hematological/biochemical parameters. TPL extraction yielded 1.3%; FI, presenting high carbohydrate and low protein content, at 0.1 mg/kg inhibited paw edema induced by carrageenan (77%, serotonin (54%, PGE2 (69% and nitric oxide (73%, and the peritonitis elicited by carrageenan (48% or fMLP (67%, being well tolerated by animals. FI exhibited potent anti-inflammatory activity, revealing to be important active component in traditionally prepared remedies to treat inflammatory states.

  7. Paramphistomum cervi: surface topography of the tegument of adult fluke.

    Science.gov (United States)

    Panyarachun, Busaba; Sobhon, Prasert; Tinikul, Yotsawan; Chotwiwatthanakun, Charoonroj; Anupunpisit, Vipavee; Anuracpreeda, Panat

    2010-06-01

    Adult Paramphistomum cervi or rumen fluke are pear-shaped, slightly concave ventrally and convex dorsally. The worm measures about 5-13 mm in length and 2-5 mm in width across the mid-section. As observed by scanning electron microscopy (SEM), the tegumental surface in all part of the body, appears highly corrugated with transverse folds alternating with grooves and is spineless. At high magnification, the surface of the fold is composed of microfolds or ridges separated by microgrooves or pits. Corrugations and invaginations of the ventral surface are also more extensive than on the dorsal surface of the body. Both anterior and posterior suckers have thick rims covered with transverse folds without spine. The genital pore is situated at the anterior third of the body. There are two types of sensory papillae on the surface: type 1 is bulbous in shape, measuring 10-15 microm in diameter at the base with nipple-like tips, and type 2 has a similar shape and size and also a short cilia on top. These sensory papillae usually occur in large clusters, each having between 5 and 20 units depending on the region of the body. Clusters of papillae on the ventral surface and around the anterior suckers tend to be more numerous and larger in size. The dorsal surface of the body has the least number of papillae. Copyright (c) 2009 Elsevier Inc. All rights reserved.

  8. Surface topography and ultrastructural architecture of the tegument of adult Carmyerius spatiosus Brandes, 1898.

    Science.gov (United States)

    Anuracpreeda, Panat; Phutong, Sumittra; Ngamniyom, Arin; Panyarachun, Busaba; Sobhon, Prasert

    2015-03-01

    Adult Carmyerius spatiosus or stomach fluke has an elongate, cylindrical-shaped, straight to slightly curved body, with conical anterior end and truncated posterior end. The worm measures about 8.7-11.2mm in body length and 2.3-3.0mm in body width across the mid-section. When observed by SEM, the tegumental surface in all part of the body appears highly corrugated with ridges and furrows, and having no spines. The ventral surface has more complex corrugation than those of the dorsal surface. Both anterior and posterior suckers have thick edges covered with transverse folds and appear spineless. The genital pore is located at the anterior part of the body. There are two types of sensory papillae on the surface: type 1 is bulbous in shape with nipple-like tips; type 2 has a similar shape with short cilia on the tip. The dorsal surface exhibits similar surface features, but papillae appear less numerous and are smaller. When observed by TEM, the tegument is divided into four layers. The first layer includes the ridges and furrows which are covered by a trilaminate membrane underlined by a dense lamina and coated externally with the glycocalyx. The second layer of the tegument is a narrow region of cytoplasm that contains high concentrations of ovoid electron lucent tegumental granules (TG1), and disc-shaped electron dense tegumental granules (TG2) as well as lysosomes. TG1 close to the surface invariably exocytose their content into bottoms of the ridges, while some TG2 are fused and have their membrane joined up with the surface membrane. The third layer is the widest middle area of the tegument which contains numerous and evenly distributed mitochondria. Both TG1 and TG2 granules are present but in much fewer number than in the first and second layers. The fourth layer is the innermost zone that rests on and couples with a thick basal lamina. The cytoplasm in this layer is loosely packed and contains numerous infoldings of the basal plasma membrane with closely

  9. Human Cytomegalovirus pUL47 Modulates Tegumentation and Capsid Accumulation at the Viral Assembly Complex

    Science.gov (United States)

    Cappadona, Ilaria; Villinger, Clarissa; Schutzius, Gabi; Mertens, Thomas

    2015-01-01

    ABSTRACT Human cytomegalovirus (HCMV) tegument protein pUL47 is an interaction partner of pUL48 and highly conserved among herpesviruses. It is closely associated with the capsid and has an important function early in infection. Here, we report a specific role of pUL47 in the tegumentation of capsids in the cytoplasm. A newly generated mutant virus (TB-47stop), in which expression of pUL47 is blocked, exhibited a severe impairment in cell-to-cell spread and release of infectivity from infected cells. Ultrastructural analysis of TB-47stop-infected cells clearly showed cytoplasmic accumulations of nonenveloped capsids that were only partially tegumented, indicating that these capsids failed to complete tegumentation. Nevertheless, these accumulations were positive for HCMV inner tegument proteins pp150 and pUL48, suggesting that their attachment to capsids occurs independently of pUL47. Despite these morphological alterations, fully enveloped virus particles were found in the extracellular space and at the viral assembly complex (vAC) of TB-47stop-infected cells, indicating that pUL47 is not essential for the generation of virions. We confirmed findings that incorporation of pUL48 into virions is impaired in the absence of pUL47. Interestingly, pUL47 exhibited a strong nuclear localization in transfected cells, whereas it was found exclusively at the vAC in the context of virus infection. Colocalization of pUL47 and pUL48 at the vAC is consistent with their interaction. We also found a shift to a more nuclear localization of pUL47 when the expression of pUL48 was reduced. Summarizing our results, we hypothesize that pUL48 directs pUL47 to the vAC to promote tegumentation and secondary envelopment of capsids. IMPORTANCE Generation of infectious HCMV particles requires an organized and multistep process involving the action of several viral and cellular proteins as well as protein-protein interactions. A better understanding of these processes is important for

  10. EBV tegument protein BNRF1 disrupts DAXX-ATRX to activate viral early gene transcription.

    Directory of Open Access Journals (Sweden)

    Kevin Tsai

    2011-11-01

    Full Text Available Productive infection by herpesviruses involve the disabling of host-cell intrinsic defenses by viral encoded tegument proteins. Epstein-Barr Virus (EBV typically establishes a non-productive, latent infection and it remains unclear how it confronts the host-cell intrinsic defenses that restrict viral gene expression. Here, we show that the EBV major tegument protein BNRF1 targets host-cell intrinsic defense proteins and promotes viral early gene activation. Specifically, we demonstrate that BNRF1 interacts with the host nuclear protein Daxx at PML nuclear bodies (PML-NBs and disrupts the formation of the Daxx-ATRX chromatin remodeling complex. We mapped the Daxx interaction domain on BNRF1, and show that this domain is important for supporting EBV primary infection. Through reverse transcription PCR and infection assays, we show that BNRF1 supports viral gene expression upon early infection, and that this function is dependent on the Daxx-interaction domain. Lastly, we show that knockdown of Daxx and ATRX induces reactivation of EBV from latently infected lymphoblastoid cell lines (LCLs, suggesting that Daxx and ATRX play a role in the regulation of viral chromatin. Taken together, our data demonstrate an important role of BNRF1 in supporting EBV early infection by interacting with Daxx and ATRX; and suggest that tegument disruption of PML-NB-associated antiviral resistances is a universal requirement for herpesvirus infection in the nucleus.

  11. EBV Tegument Protein BNRF1 Disrupts DAXX-ATRX to Activate Viral Early Gene Transcription

    Science.gov (United States)

    Tsai, Kevin; Thikmyanova, Nadezhda; Wojcechowskyj, Jason A.; Delecluse, Henri-Jacques; Lieberman, Paul M.

    2011-01-01

    Productive infection by herpesviruses involve the disabling of host-cell intrinsic defenses by viral encoded tegument proteins. Epstein-Barr Virus (EBV) typically establishes a non-productive, latent infection and it remains unclear how it confronts the host-cell intrinsic defenses that restrict viral gene expression. Here, we show that the EBV major tegument protein BNRF1 targets host-cell intrinsic defense proteins and promotes viral early gene activation. Specifically, we demonstrate that BNRF1 interacts with the host nuclear protein Daxx at PML nuclear bodies (PML-NBs) and disrupts the formation of the Daxx-ATRX chromatin remodeling complex. We mapped the Daxx interaction domain on BNRF1, and show that this domain is important for supporting EBV primary infection. Through reverse transcription PCR and infection assays, we show that BNRF1 supports viral gene expression upon early infection, and that this function is dependent on the Daxx-interaction domain. Lastly, we show that knockdown of Daxx and ATRX induces reactivation of EBV from latently infected lymphoblastoid cell lines (LCLs), suggesting that Daxx and ATRX play a role in the regulation of viral chromatin. Taken together, our data demonstrate an important role of BNRF1 in supporting EBV early infection by interacting with Daxx and ATRX; and suggest that tegument disruption of PML-NB-associated antiviral resistances is a universal requirement for herpesvirus infection in the nucleus. PMID:22102817

  12. Carbohydrate Detection and Lectin Isolation from Tegumental Tissue of Fasciola hepatica

    Directory of Open Access Journals (Sweden)

    MB Molaei Rad

    2010-02-01

    Full Text Available "nBackground: Fascioliasis is a chronic hepatic disease and may be resulted from mechani­cal/molecular parasite adhesion to host liver tissue. The aim of this study was to detect surface car­bohydrate and lectin, carbohydrate-binding protein isolation that might be responsible of this molecular binding."nMethods: The present experimental work was conducted in the Department of Medical Parasitol­ogy and Mycology, School of Public Health, Tehran University of Medical Sciences, Te­hran, Iran.  Fasciola hepatica parasites were collected from abattoir (Saman, Tehran, Iran and surface mannose-carbohydrate was detected by fluorescein isothiocyanate (FITC conju­gated lectin (Lentil. Lectin of tegumental tissue from F. hepatica was isolated by affinity chroma­tography and detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE."nResults: Mannose carbohydrate was observed on the surface of tegumental tissue from para­site under fluorescence microscope. Carbohydrate-binding protein or lectin with MW of 50 kDa also was isolated from homogenized tegument of helminth."nConclusion: These results are important for understanding of molecular pathogenesis of F. hepat­ica at the chronic phase of fascioliasis

  13. Molecular and Structural Characterization of the Tegumental 20.6-kDa Protein in Clonorchis sinensis as a Potential Druggable Target

    OpenAIRE

    Kim, Yu-Jung; Yoo, Won Gi; Lee, Myoung-Ro; Kang, Jung-Mi; Na, Byoung-Kuk; Cho, Shin-Hyeong; Park, Mi-Yeoun; Ju, Jung-Won

    2017-01-01

    The tegument, representing the membrane-bound outer surface of platyhelminth parasites, plays an important role for the regulation of the host immune response and parasite survival. A comprehensive understanding of tegumental proteins can provide drug candidates for use against helminth-associated diseases, such as clonorchiasis caused by the liver fluke Clonorchis sinensis. However, little is known regarding the physicochemical properties of C. sinensis teguments. In this study, a novel 20.6...

  14. Scanning and transmission electron microscopy of the tegument of Paranaella luquei Kohn, Baptista-Farias & Cohen, 2000 (Microcotylidae, Monogenea, parasite of a Brazilian catfish, Hypostomus regani

    Directory of Open Access Journals (Sweden)

    SC Cohen

    2001-05-01

    Full Text Available The surface topography and ultrastructure of the tegument of Paranaella luquei Kohn, Baptista-Farias & Cohen, 2000, a microcotylid monogenean parasite from the gills of Hypostomus regani (Ihering, 1905 (Loricariidae was studied by scanning (SEM and transmission electron microscopy (TEM. By SEM, it was observed that the tegument presents transversal ridges, forming folds in the ventral and dorsal surfaces and microvillous-like tegumental projections in the anterior and median regions of body. These projections were also observed by TEM. The tegument is made up of a syncytium delimited by apical and basal plasma membranes, containing inclusion bodies and mitochondria, connected to the nucleated region by means of cytoplasmatic processes. The tegumental cells present a well developed nucleus and cytoplasm containing inclusion bodies, similar to those found on the external layer, mitochondria, rough endoplasmatic reticulum and free ribossomes.

  15. Identification of interaction domains within the UL37 tegument protein of herpes simplex virus type 1.

    Science.gov (United States)

    Bucks, Michelle A; Murphy, Michael A; O'Regan, Kevin J; Courtney, Richard J

    2011-07-20

    Herpes simplex virus type 1 (HSV-1) UL37 is a 1123 amino acid tegument protein that self-associates and binds to the tegument protein UL36 (VP1/2). Studies were undertaken to identify regions of UL37 involved in these protein-protein interactions. Coimmunoprecipitation assays showed that residues within the carboxy-terminal half of UL37, amino acids 568-1123, are important for interaction with UL36. Coimmunoprecipitation assays also revealed that amino acids 1-300 and 568-1123 of UL37 are capable of self-association. UL37 appears to self-associate only under conditions when UL36 is not present or is present in low amounts, suggesting UL36 and UL37 may compete for binding. Transfection-infection experiments were performed to identify domains of UL37 that complement the UL37 deletion virus, K∆UL37. The carboxy-terminal region of UL37 (residues 568-1123) partially rescues the K∆UL37 infection. These results suggest the C-terminus of UL37 may contribute to its essential functional role within the virus-infected cell. Copyright © 2011 Elsevier Inc. All rights reserved.

  16. Time-dependent tegumental surface changes in juvenile Fasciola gigantica in response to triclabendazole treatment in goat.

    Science.gov (United States)

    Shareef, P A Ahammed; Brennan, Gerard P; McVeigh, Paul; Khan, M A Hannan; Morphew, Russell M; Mousley, Angela; Marks, Nikki J; Saifullah, M K; Brophy, Peter M; Maule, Aaron G; Abidi, S M A

    2014-08-01

    Triclabendazole (TCBZ), the anthelmintic drug active against both mature and immature liver flukes, was used to investigate the effect of in vivo treatment on the tegumental surface of juvenile Fasciola gigantica. Five goats were infected with 150 F. gigantica metacercariae each by oral gavage. Four of them were treated with single dose of TCBZ at 10mg/kg at four weeks post-infection. They were euthanized at 0 (untreated), 24, 48, 72 and 96h post treatment. Juvenile flukes were manually retrieved from the goat livers and processed for scanning electron microscopy. In control flukes, the anterior region was adorned with sharply pointed spines projecting away from the surface, while in the posterior region, spines become shorter and narrower, loosing serration and with the appearance of distinct furrows and papillae. The dorsal surface retained the same pattern of surface architecture similar to that of ventral surface. Flukes obtained from 24h post-treatment did not show any apparent change and were still very active. However, there were limited movements and some blebbing, swelling, deposition of tegumental secretions and some flattening displayed by the flukes of 48h post-treatment. All the worms were found dead 72h post-treatment and showed advanced level of tegumental disruptions, consisting of severe distortion of spines, sloughing off the tegument to expose the basal lamina, formation of pores and isolated patches of lesions. By 96h post-treatment, the disruption was extremely severe and the tegument was completely sheared off causing deeper lesions that exposed the underlying musculature. The disruption was more severe at posterior than anterior region and on ventral than dorsal surface. The present study further establishes the time-course of TCBZ action in vivo with 100% efficacy against the juvenile tropical liver fluke. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Gammaherpesviral Tegument Proteins, PML-Nuclear Bodies and the Ubiquitin-Proteasome System

    Directory of Open Access Journals (Sweden)

    Florian Full

    2017-10-01

    Full Text Available Gammaherpesviruses like Epstein-Barr virus (EBV and Kaposi’s sarcoma-associated herpesvirus (KSHV subvert the ubiquitin proteasome system for their own benefit in order to facilitate viral gene expression and replication. In particular, viral tegument proteins that share sequence homology to the formylglycineamide ribonucleotide amidotransferase (FGARAT, or PFAS, an enzyme in the cellular purine biosynthesis, are important for disrupting the intrinsic antiviral response associated with Promyelocytic Leukemia (PML protein-associated nuclear bodies (PML-NBs by proteasome-dependent and independent mechanisms. In addition, all herpesviruses encode for a potent ubiquitin protease that can efficiently remove ubiquitin chains from proteins and thereby interfere with several different cellular pathways. In this review, we discuss mechanisms and functional consequences of virus-induced ubiquitination and deubiquitination for early events in gammaherpesviral infection.

  18. Identification of host cell proteins which interact with herpes simplex virus type 1 tegument protein pUL37.

    Science.gov (United States)

    Kelly, Barbara J; Diefenbach, Eve; Fraefel, Cornel; Diefenbach, Russell J

    2012-01-20

    The herpes simplex virus type 1 (HSV-1) structural tegument protein pUL37, which is conserved across the Herpesviridae family, is known to be essential for secondary envelopment during the egress of viral particles. To shed light on additional roles of pUL37 during viral replication a yeast two-hybrid screen of a human brain cDNA library was undertaken. This screen identified ten host cell proteins as potential pUL37 interactors. One of the interactors, serine threonine kinase TAOK3, was subsequently confirmed to interact with pUL37 using an in vitro pulldown assay. Such host cell/pUL37 interactions provide further insights into the multifunctional role of this herpesviral tegument protein. Copyright © 2011 Elsevier Inc. All rights reserved.

  19. Interaction and interdependent packaging of tegument protein UL11 and glycoprotein e of herpes simplex virus.

    Science.gov (United States)

    Han, Jun; Chadha, Pooja; Meckes, David G; Baird, Nicholas L; Wills, John W

    2011-09-01

    The UL11 tegument protein of herpes simplex virus plays a critical role in the secondary envelopment; however, the mechanistic details remain elusive. Here, we report a new function of UL11 in the budding process in which it directs efficient acquisition of glycoprotein E (gE) via a direct interaction. In vitro binding assays showed that the interaction required only the first 28, membrane-proximal residues of the cytoplasmic tail of gE, and the C-terminal 26 residues of UL11. A second, weaker binding site was also found in the N-terminal half of UL11. The significance of the gE-UL11 interaction was subsequently investigated with viral deletion mutants. In the absence of the gE tail, virion packaging of UL11, but not other tegument proteins such as VP22 and VP16, was reduced by at least 80%. Reciprocally, wild-type gE packaging was also drastically reduced by about 87% in the absence of UL11, and this defect could be rescued in trans by expressing U(L)11 at the U(L)35 locus. Surprisingly, a mutant that lacks the C-terminal gE-binding site of UL11 packaged nearly normal amounts of gE despite its strong interaction with the gE tail in vitro, indicating that the interaction with the UL11 N terminus may be important. Mutagenesis studies of the UL11 N terminus revealed that the association of UL11 with membrane was not required for this function. In contrast, the UL11 acidic cluster motif was found to be critical for gE packaging and was not replaceable with foreign acidic clusters. Together, these results highlight an important role of UL11 in the acquisition of glycoprotein-enriched lipid bilayers, and the findings may also have important implications for the role of UL11 in gE-mediated cell-to-cell spread.

  20. Fasciola gigantica enolase is a major component of worm tegumental fraction protective against sheep fasciolosis.

    Science.gov (United States)

    Mahana, N; Abd-Allah, H A-S; Salah, M; Tallima, H; El Ridi, R

    2016-06-01

    Infection of cattle and sheep with the parasite Fasciola gigantica is a cause of important economic losses throughout Asia and Africa. Many of the available anthelmintics have undesirable side effects, and the parasite may acquire drug resistance as a result of mass and repeated treatments of livestock. Accordingly, the need for developing a vaccine is evident. Triton-soluble surface membrane and tegumental proteins (TSMTP) of 60, 32, and 28 kDa previously shown to elicit protective immunity in mice against challenge F. gigantica infection were found to be strongly immunogenic in sheep eliciting vigorous specific antibody responses to a titer>1:16,000 as assessed by enzyme-linked immunosorbent assay. Furthermore, the 60 kDa fraction induced production of antibodies able to bind to the surface membrane of newly excysted juvenile flukes and mediate their attrition in antibody-dependent complement- and cell-mediated cytotoxicity assays, and significant (PFasciola hepatica enolase, suggesting that a fasciolosis vaccine might be effective against both tropical and temperate liver flukes. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Evaluation and Characterization of Fasciola hepatica Tegument Protein Extract for Serodiagnosis of Human Fascioliasis

    Science.gov (United States)

    Morales, Adelaida

    2012-01-01

    Tegument protein extract from Fasciola hepatica adult flukes (FhTA) was obtained and assessed for its potential as a diagnostic agent for the serological detection of human fascioliasis using an indirect enzyme-linked immunosorbent assay (ELISA). In an analysis of sera from 45 patients infected with F. hepatica, sera from 41 patients with other parasitic infections, and sera from 33 healthy controls, the FhTA-ELISA showed sensitivity, specificity, and accuracy of 91.1%, 97.3%, and 95%, respectively. Specific IgG1 and IgG4 were the antibody isotypes mainly detected in sera from patients with fascioliasis. Polypeptides of 52, 38, 24 to 26, and 12 to 14 kDa were identified by Western blotting as the most immunoreactive components of the FhTA. A proteomic approach led us to identify enolase, aldolase, glutathione S-transferase, and fatty acid binding protein as the major immunoreactive components of the FhTA. PMID:23015645

  2. Enzymatic shaving of the tegument surface of live schistosomes for proteomic analysis: a rational approach to select vaccine candidates.

    Directory of Open Access Journals (Sweden)

    William Castro-Borges

    2011-03-01

    Full Text Available The membrane-associated and membrane-spanning constituents of the Schistosoma mansoni tegument surface, the parasite's principal interface with the host bloodstream, have recently been characterized using proteomic techniques. Biotinylation of live worms using membrane-impermeant probes revealed that only a small subset of the proteins was accessible to the reagents. Their position within the multilayered architecture of the surface has not been ascertained.An enzymatic shaving approach on live worms has now been used to release the most accessible components, for analysis by MS/MS. Treatment with trypsin, or phosphatidylinositol-specific phospholipase C (PiPLC, only minimally impaired membrane integrity. PiPLC-enriched proteins were distinguished from those released in parasite vomitus or by handling damage, using isobaric tagging. Trypsin released five membrane proteins, Sm200, Sm25 and three annexins, plus host CD44 and the complement factors C3 and C4. Nutrient transporters and ion channels were absent from the trypsin fraction, suggesting a deeper location in the surface complex; surprisingly, two BAR-domain containing proteins were released. Seven parasite and two host proteins were enriched by PiPLC treatment, the vaccine candidate Sm29 being the most prominent along with two orthologues of human CD59, potentially inhibitors of complement fixation. The enzymes carbonic anhydrase and APD-ribosyl cyclase were also enriched, plus Sm200 and alkaline phosphatase. Host GPI-anchored proteins CD48 and CD90, suggest 'surface painting' during worm peregrination in the portal system.Our findings suggest that the membranocalyx secreted over the tegument surface is not the inert barrier previously proposed, some tegument proteins being externally accessible to enzymes and thus potentially located within it. Furthermore, the detection of C3 and C4 indicates that the complement cascade is initiated, while two CD59 orthologues suggest a potential

  3. Plus- and minus-end directed microtubule motors bind simultaneously to herpes simplex virus capsids using different inner tegument structures.

    Directory of Open Access Journals (Sweden)

    Kerstin Radtke

    2010-07-01

    Full Text Available Many viruses depend on host microtubule motors to reach their destined intracellular location. Viral particles of neurotropic alphaherpesviruses such as herpes simplex virus 1 (HSV1 show bidirectional transport towards the cell center as well as the periphery, indicating that they utilize microtubule motors of opposing directionality. To understand the mechanisms of specific motor recruitment, it is necessary to characterize the molecular composition of such motile viral structures. We have generated HSV1 capsids with different surface features without impairing their overall architecture, and show that in a mammalian cell-free system the microtubule motors dynein and kinesin-1 and the dynein cofactor dynactin could interact directly with capsids independent of other host factors. The capsid composition and surface was analyzed with respect to 23 structural proteins that are potentially exposed to the cytosol during virus assembly or cell entry. Many of these proteins belong to the tegument, the hallmark of all herpesviruses located between the capsid and the viral envelope. Using immunoblots, quantitative mass spectrometry and quantitative immunoelectron microscopy, we show that capsids exposing inner tegument proteins such as pUS3, pUL36, pUL37, ICP0, pUL14, pUL16, and pUL21 recruited dynein, dynactin, kinesin-1 and kinesin-2. In contrast, neither untegumented capsids exposing VP5, VP26, pUL17 and pUL25 nor capsids covered by outer tegument proteins such as vhs, pUL11, ICP4, ICP34.5, VP11/12, VP13/14, VP16, VP22 or pUS11 bound microtubule motors. Our data suggest that HSV1 uses different structural features of the inner tegument to recruit dynein or kinesin-1. Individual capsids simultaneously accommodated motors of opposing directionality as well as several copies of the same motor. Thus, these associated motors either engage in a tug-of-war or their activities are coordinately regulated to achieve net transport either to the nucleus during

  4. Molecular and Structural Characterization of the Tegumental 20.6-kDa Protein in Clonorchis sinensis as a Potential Druggable Target

    Directory of Open Access Journals (Sweden)

    Yu-Jung Kim

    2017-03-01

    Full Text Available The tegument, representing the membrane-bound outer surface of platyhelminth parasites, plays an important role for the regulation of the host immune response and parasite survival. A comprehensive understanding of tegumental proteins can provide drug candidates for use against helminth-associated diseases, such as clonorchiasis caused by the liver fluke Clonorchis sinensis. However, little is known regarding the physicochemical properties of C. sinensis teguments. In this study, a novel 20.6-kDa tegumental protein of the C. sinensis adult worm (CsTegu20.6 was identified and characterized by molecular and in silico methods. The complete coding sequence of 525 bp was derived from cDNA clones and encodes a protein of 175 amino acids. Homology search using BLASTX showed CsTegu20.6 identity ranging from 29% to 39% with previously-known tegumental proteins in C. sinensis. Domain analysis indicated the presence of a calcium-binding EF-hand domain containing a basic helix-loop-helix structure and a dynein light chain domain exhibiting a ferredoxin fold. We used a modified method to obtain the accurate tertiary structure of the CsTegu20.6 protein because of the unavailability of appropriate templates. The CsTegu20.6 protein sequence was split into two domains based on the disordered region, and then, the structure of each domain was modeled using I-TASSER. A final full-length structure was obtained by combining two structures and refining the whole structure. A refined CsTegu20.6 structure was used to identify a potential CsTegu20.6 inhibitor based on protein structure-compound interaction analysis. The recombinant proteins were expressed in Escherichia coli and purified by nickel-nitrilotriacetic acid affinity chromatography. In C. sinensis, CsTegu20.6 mRNAs were abundant in adult and metacercariae, but not in the egg. Immunohistochemistry revealed that CsTegu20.6 localized to the surface of the tegument in the adult fluke. Collectively, our results

  5. Molecular and Structural Characterization of the Tegumental 20.6-kDa Protein in Clonorchis sinensis as a Potential Druggable Target.

    Science.gov (United States)

    Kim, Yu-Jung; Yoo, Won Gi; Lee, Myoung-Ro; Kang, Jung-Mi; Na, Byoung-Kuk; Cho, Shin-Hyeong; Park, Mi-Yeoun; Ju, Jung-Won

    2017-03-04

    The tegument, representing the membrane-bound outer surface of platyhelminth parasites, plays an important role for the regulation of the host immune response and parasite survival. A comprehensive understanding of tegumental proteins can provide drug candidates for use against helminth-associated diseases, such as clonorchiasis caused by the liver fluke Clonorchis sinensis . However, little is known regarding the physicochemical properties of C. sinensis teguments. In this study, a novel 20.6-kDa tegumental protein of the C. sinensis adult worm (CsTegu20.6) was identified and characterized by molecular and in silico methods. The complete coding sequence of 525 bp was derived from cDNA clones and encodes a protein of 175 amino acids. Homology search using BLASTX showed CsTegu20.6 identity ranging from 29% to 39% with previously-known tegumental proteins in C. sinensis . Domain analysis indicated the presence of a calcium-binding EF-hand domain containing a basic helix-loop-helix structure and a dynein light chain domain exhibiting a ferredoxin fold. We used a modified method to obtain the accurate tertiary structure of the CsTegu20.6 protein because of the unavailability of appropriate templates. The CsTegu20.6 protein sequence was split into two domains based on the disordered region, and then, the structure of each domain was modeled using I-TASSER. A final full-length structure was obtained by combining two structures and refining the whole structure. A refined CsTegu20.6 structure was used to identify a potential CsTegu20.6 inhibitor based on protein structure-compound interaction analysis. The recombinant proteins were expressed in Escherichia coli and purified by nickel-nitrilotriacetic acid affinity chromatography. In C. sinensis , CsTegu20.6 mRNAs were abundant in adult and metacercariae, but not in the egg. Immunohistochemistry revealed that CsTegu20.6 localized to the surface of the tegument in the adult fluke. Collectively, our results contribute to a

  6. A novel ex vivo immunoproteomic approach characterising Fasciola hepatica tegumental antigens identified using immune antibody from resistant sheep.

    Science.gov (United States)

    Cameron, Timothy C; Cooke, Ira; Faou, Pierre; Toet, Hayley; Piedrafita, David; Young, Neil; Rathinasamy, Vignesh; Beddoe, Travis; Anderson, Glenn; Dempster, Robert; Spithill, Terry W

    2017-08-01

    A more thorough understanding of the immunological interactions between Fasciola spp. and their hosts is required if we are to develop new immunotherapies to control fasciolosis. Deeper knowledge of the antigens that are the target of the acquired immune responses of definitive hosts against both Fasciola hepatica and Fasciola gigantica will potentially identify candidate vaccine antigens. Indonesian Thin Tail sheep express a high level of acquired immunity to infection by F. gigantica within 4weeks of infection and antibodies in Indonesian Thin Tail sera can promote antibody-dependent cell-mediated cytotoxicity against the surface tegument of juvenile F. gigantica in vitro. Given the high protein sequence similarity between F. hepatica and F. gigantica, we hypothesised that antibody from F. gigantica-infected sheep could be used to identify the orthologous proteins in the tegument of F. hepatica. Purified IgG from the sera of F. gigantica-infected Indonesian Thin Tail sheep collected pre-infection and 4weeks p.i. were incubated with live adult F. hepatica ex vivo and the immunosloughate (immunoprecipitate) formed was isolated and analysed via liquid chromatography-electrospray ionisation-tandem mass spectrometry to identify proteins involved in the immune response. A total of 38 proteins were identified at a significantly higher abundance in the immunosloughate using week 4 IgG, including eight predicted membrane proteins, 20 secreted proteins, nine proteins predicted to be associated with either the lysosomes, the cytoplasm or the cytoskeleton and one protein with an unknown cellular localization. Three of the membrane proteins are transporters including a multidrug resistance protein, an amino acid permease and a glucose transporter. Interestingly, a total of 21 of the 38 proteins matched with proteins recently reported to be associated with the proposed small exosome-like extracellular vesicles of adult F. hepatica, suggesting that the Indonesian Thin Tail week

  7. Schistosome tegumental ecto-apyrase (SmATPDase1 degrades exogenous pro-inflammatory and pro-thrombotic nucleotides

    Directory of Open Access Journals (Sweden)

    Akram A. Da’dara

    2014-03-01

    Full Text Available Schistosomes are parasitic worms that can survive in the hostile environment of the human bloodstream where they appear refractory to both immune elimination and thrombus formation. We hypothesize that parasite migration in the bloodstream can stress the vascular endothelium causing this tissue to release chemicals alerting responsive host cells to the stress. Such chemicals are called damage associated molecular patterns (DAMPs and among the most potent is the proinflammatory mediator, adenosine triphosphate (ATP. Furthermore, the ATP derivative ADP is a pro-thrombotic molecule that acts as a strong activator of platelets. Schistosomes are reported to possess at their host interactive tegumental surface a series of enzymes that could, like their homologs in mammals, degrade extracellular ATP and ADP. These are alkaline phosphatase (SmAP, phosphodiesterase (SmNPP-5 and ATP diphosphohydrolase (SmATPDase1. In this work we employ RNAi to knock down expression of the genes encoding these enzymes in the intravascular life stages of the parasite. We then compare the abilities of these parasites to degrade exogenously added ATP and ADP. We find that only SmATPDase1-suppressed parasites are significantly impaired in their ability to degrade these nucleotides. Suppression of SmAP or SmNPP-5 does not appreciably affect the worms’ ability to catabolize ATP or ADP. These findings are confirmed by the functional characterization of the enzymatically active, full-length recombinant SmATPDase1 expressed in CHO-S cells. The enzyme is a true apyrase; SmATPDase1 degrades ATP and ADP in a cation dependent manner. Optimal activity is seen at alkaline pH. The Km of SmATPDase1 for ATP is 0.4 ± 0.02 mM and for ADP, 0.252 ± 0.02 mM. The results confirm the role of tegumental SmATPDase1 in the degradation of the exogenous pro-inflammatory and pro-thrombotic nucleotides ATP and ADP by live intravascular stages of the parasite. By degrading host inflammatory signals

  8. Virus-like particles vaccine containing Clonorchis sinensis tegumental protein induces partial protection against Clonorchis sinensis infection.

    Science.gov (United States)

    Lee, Dong-Hun; Kim, Ah-Ra; Lee, Su-Hwa; Quan, Fu-Shi

    2017-12-29

    Human clonorchiasis, caused by the infection of Clonorchis sinensis, is one of the major health problems in Southeast Asia. However, vaccine efficacy against C. sinensis infection remains largely unknown. In this study, for the first time, we generated virus-like particles (VLPs) vaccine containing the C. sinensis tegumental protein 22.3 kDa (CsTP 22.3) and the influenza matrix protein (M1) as a core protein, and investigated the vaccine efficacy in Sprague-Dawley rats. Intranasal immunization of VLPs vaccine induced C. sinensis-specific IgG, IgG2a and IgG2c in the sera and IgA responses in the feces and intestines. Notably, upon challenge infection with C. sinensis metacercariae, significantly lower adult worm loads (70.2%) were measured in the liver of rats immunized with VLPs, compared to those of naïve rats. Furthermore, VLPs immunization induced antibody secreting cells (ASC) responses and CD4+/CD8+ T cell responses in the spleen. Our results indicated that VLPs vaccine containing C. sinensis CsTP 22.3 kDa provided partial protection against C. sisnensis infection. Thus, VLPs could be a potential vaccine candidate against C. sinensis.

  9. Schistosoma japonicum UDP-glucose 4-epimerase protein is located on the tegument and induces moderate protection against challenge infection.

    Directory of Open Access Journals (Sweden)

    Pingping Liu

    Full Text Available Schistosomiasis is an important global public health problem, as millions of people are at risk of acquiring this infection. An ideal method for sustainable control of schistosomiasis is using a vaccine alone or in combination with drugs. In the present study, we cloned the SjGALE gene and generated the expression product in E. coli. The expression level of SjGALE during different developmental stages of S. japonicum was evaluated by real-time RT-PCR and western blotting. Immunolocalization indicated that the protein was mainly located on the tegument of the parasite. Infection of rSjGALE-immunized mice demonstrated a 34% and 49% reduction of the mean worm burden and liver egg burden, respectively, in two independent experiments, indicating immune protection. The liver egg count from each female adult worm was significantly reduced by 63% in the two trials. The cytokine profile and IgG isotype analysis demonstrated the induction of a Th1 immune profile in response to immunization with this protein, further suggesting protection against infection. In conclusion, these findings indicated that SjGALE is a potential vaccine against S. japonicum.

  10. Inner tegument proteins of Herpes Simplex Virus are sufficient for intracellular capsid motility in neurons but not for axonal targeting

    Science.gov (United States)

    Müller, Oliver; Ivanova, Lyudmila; Bialy, Dagmara; Pohlmann, Anja; Binz, Anne; Hegemann, Maike; Viejo-Borbolla, Abel; Rosenhahn, Bodo; Bauerfeind, Rudolf; Sodeik, Beate

    2017-01-01

    Upon reactivation from latency and during lytic infections in neurons, alphaherpesviruses assemble cytosolic capsids, capsids associated with enveloping membranes, and transport vesicles harboring fully enveloped capsids. It is debated whether capsid envelopment of herpes simplex virus (HSV) is completed in the soma prior to axonal targeting or later, and whether the mechanisms are the same in neurons derived from embryos or from adult hosts. We used HSV mutants impaired in capsid envelopment to test whether the inner tegument proteins pUL36 or pUL37 necessary for microtubule-mediated capsid transport were sufficient for axonal capsid targeting in neurons derived from the dorsal root ganglia of adult mice. Such neurons were infected with HSV1-ΔUL20 whose capsids recruited pUL36 and pUL37, with HSV1-ΔUL37 whose capsids associate only with pUL36, or with HSV1-ΔUL36 that assembles capsids lacking both proteins. While capsids of HSV1-ΔUL20 were actively transported along microtubules in epithelial cells and in the somata of neurons, those of HSV1-ΔUL36 and -ΔUL37 could only diffuse in the cytoplasm. Employing a novel image analysis algorithm to quantify capsid targeting to axons, we show that only a few capsids of HSV1-ΔUL20 entered axons, while vesicles transporting gD utilized axonal transport efficiently and independently of pUL36, pUL37, or pUL20. Our data indicate that capsid motility in the somata of neurons mediated by pUL36 and pUL37 does not suffice for targeting capsids to axons, and suggest that capsid envelopment needs to be completed in the soma prior to targeting of herpes simplex virus to the axons, and to spreading from neurons to neighboring cells. PMID:29284065

  11. Immunization with tegument nucleotidases associated with a subcurative praziquantel treatment reduces worm burden following Schistosoma mansoni challenge

    Directory of Open Access Journals (Sweden)

    Henrique K. Rofatto

    2013-04-01

    Full Text Available Schistosomiasis is a debilitating disease caused by flatworm parasites of the Schistosoma genus and remains a high public health impact disease around the world, although effective treatment with Praziquantel (PZQ has been available since the 1970s. Control of this disease would be greatly improved by the development of a vaccine, which could be combined with chemotherapy. The sequencing of the Schistosoma mansoni transcriptome and genome identified a range of potential vaccine antigens. Among these, three nucleotidases from the tegument of the parasite, presumably involved in purinergic signaling and nucleotide metabolism, were proposed as promising vaccine candidates: an alkaline phosphatase (SmAP, a phosphodiesterase (SmNPP-5 and a diphosphohydrolase (SmNTPDase. Herein, we evaluate the potential of these enzymes as vaccine antigens, with or without subcurative PZQ treatment. Immunization of mice with the recombinant proteins alone or in combination demonstrated that SmAP is the most immunogenic of the three. It induced the highest antibody levels, particularly IgG1, associated with an inflammatory cellular immune response characterized by high TNF-α and a Th17 response, with high IL-17 expression levels. Despite the specific immune response induced, immunization with the isolated or combined proteins did not reduce the worm burden of challenged mice. Nonetheless, immunization with SmAP alone or with the three proteins combined, together with subcurative PZQ chemotherapy was able to reduce the worm burden by around 40%. The immunogenicity and relative exposure of SmAP to the host immune system are discussed, as key factors involved in the apparently synergistic effect of SmAP immunization and subcurative PZQ treatment.

  12. Novel Structure and Unexpected RNA-Binding Ability of the C-Terminal Domain of Herpes Simplex Virus 1 Tegument Protein UL21

    Energy Technology Data Exchange (ETDEWEB)

    Metrick, Claire M.; Heldwein, Ekaterina E. (Tufts-MED)

    2016-04-06

    Proteins forming the tegument layers of herpesviral virions mediate many essential processes in the viral replication cycle, yet few have been characterized in detail. UL21 is one such multifunctional tegument protein and is conserved among alphaherpesviruses. While UL21 has been implicated in many processes in viral replication, ranging from nuclear egress to virion morphogenesis to cell-cell spread, its precise roles remain unclear. Here we report the 2.7-Å crystal structure of the C-terminal domain of herpes simplex virus 1 (HSV-1) UL21 (UL21C), which has a unique α-helical fold resembling a dragonfly. Analysis of evolutionary conservation patterns and surface electrostatics pinpointed four regions of potential functional importance on the surface of UL21C to be pursued by mutagenesis. In combination with the previously determined structure of the N-terminal domain of UL21, the structure of UL21C provides a 3-dimensional framework for targeted exploration of the multiple roles of UL21 in the replication and pathogenesis of alphaherpesviruses. Additionally, we describe an unanticipated ability of UL21 to bind RNA, which may hint at a yet unexplored function.

    IMPORTANCEDue to the limited genomic coding capacity of viruses, viral proteins are often multifunctional, which makes them attractive antiviral targets. Such multifunctionality, however, complicates their study, which often involves constructing and characterizing null mutant viruses. Systematic exploration of these multifunctional proteins requires detailed road maps in the form of 3-dimensional structures. In this work, we determined the crystal structure of the C-terminal domain of UL21, a multifunctional tegument protein that is conserved among alphaherpesviruses. Structural analysis pinpointed surface areas of potential functional importance that provide a starting point for mutagenesis. In addition, the unexpected RNA-binding ability of UL21 may expand its functional repertoire

  13. Tegumental Ca-stimulated adenosine triphosphatase activity in adult Schistosoma mansoni worms Atividade da adenosina trifosfatase estimulada pelo Ca no tegumento de vermes adultos de Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    Italo M. Cesari

    1989-09-01

    Full Text Available A Ca-stimulated ATPase activity (pH 9.5 associated with the tegumental membrane enriched (TME fraction of Schistosoma mansoni adults was partially inhibited by NAP-taurine or by increasing concentrations of chlorpromazine; endogenous calmodulin was found associated with the TME fraction. A similar activity (pH 8.6 was histochemically visualized whithin the tegument of fixed worms on the cytoplasmic leaflet of both the doubel surface membrane and the basement membrane; this reaction was inhibited by 1 µM chloropromazine and it was also observed on the inner side of double membrane vesicles present in the TME fraction. No ATPase activity could be seen at alkaline pH with added Mg or Na/K ions. Without ATP, the addition of external Ca to the fixed worms induced the appearance of lead precipitates on the tegumental discoid bodies; this reaction was inhibited by molybdate and not by chlorpromazine. The intrategumentary regulation of calcium by the systems described and the possible use of phenothiazines against schistosimes are discussed.A atividade ATPse (pH 9.5 estimulada por ions de Ca associados a uma fração enriquecida de membranas do tegumento (fração EMT de vermes adultos de Schistosoma mansoni, foi inibida pro NAP-taurina ou por concentrações crescentes de clorpromacina. Foi encontrada calmodulina enfogena associada principlamente a esta fração. Em vermes adultos fixados com glutaraldeido se detectou histoquimicamente uma atividade ATPase similar (pH 8.6 na face citoplasmática da dupla membrana de superfície e da membrana por 1 µM de clorpromacina e foi também observada na face interna de vesículas de dupla membrana presentes na fração EMT. Não se pôde detectar atividade ATpase em pH alcalino na presença de ions de Mg ou Na/K. A adição externa de Ca, sem ATP, aos vermes fixados induz ao aparecimento de precipitados nos corpos discóides do tegumento; esta reação foi inibida. Os resultados são discutidos em relação a

  14. Assessment of a DNA vaccine encoding an anchored-glycosylphosphatidylinositol tegumental antigen complexed to protamine sulphate on immunoprotection against murine schistosomiasis

    Directory of Open Access Journals (Sweden)

    Eduardo JM Nascimento

    2007-02-01

    Full Text Available Protamine sulphate/DNA complexes have been shown to protect DNA from DNase digestion in a lipid system for gene transfer. A DNA-based vaccine complexed to protamine sulphate was used to induce an immune response against Schistosoma mansoni anchored-glycosylphosphatidylinositol tegumental antigen in BALB/c mice. The protection elicited ranged from 33 to 44%. The spectrum of the elicited immune response induced by the vaccine formulation without protamine was characterized by a high level of IgG (IgG1> IgG2a. Protamine sulphate added to the DNA vaccine formulation retained the green fluorescent protein encoding-plasmid longer in muscle and spleen. The experiments in vivo showed that under protamine sulphate effect, the scope of protection remained unchanged, but a modulation in antibody production (IgG1= IgG2a was observed.

  15. Herpes simplex virus type 1 tegument protein VP22 interacts with TAF-I proteins and inhibits nucleosome assembly but not regulation of histone acetylation by INHAT.

    Science.gov (United States)

    van Leeuwen, Hans; Okuwaki, Mitsuru; Hong, Rui; Chakravarti, Debabrata; Nagata, Kyosuke; O'Hare, Peter

    2003-09-01

    Affinity chromatography was used to identify cellular proteins that interact with the herpes simplex virus (HSV) tegument protein VP22. Among a small set of proteins that bind specifically to VP22, we identified TAF-I (template-activating factor I), a chromatin remodelling protein and close homologue of the histone chaperone protein NAP-1. TAF-I has been shown previously to promote more ordered transfer of histones to naked DNA through a direct interaction with histones. TAF-I, as a subunit of the INHAT (inhibitor of acetyltransferases) protein complex, also binds to histones and masks them from being substrates for the acetyltransferases p300 and PCAF. Using in vitro assays for TAF-I activity in chromatin assembly, we show that VP22 inhibits nucleosome deposition on DNA by binding to TAF-I. We also observed that VP22 binds non-specifically to DNA, an activity that is abolished by TAF-I. However, the presence of VP22 does not affect the property of INHAT in inhibiting the histone acetyltransferase activity of p300 or PCAF in vitro. We speculate that this interaction could be relevant to HSV DNA organization early in infection, for example, by interfering with nucleosomal deposition on the genome. Consistent with this possibility was the observation that overexpression of TAF-I in transfected cells interferes with the progression of HSV-1 infection.

  16. Distinct functional domains within the acidic cluster of tegument protein pp28 required for trafficking and cytoplasmic envelopment of human cytomegalovirus.

    Science.gov (United States)

    Seo, Jun-Young; Jeon, Hyejin; Hong, Sookyung; Britt, William J

    2016-10-01

    Human cytomegalovirus UL99-encoded tegument protein pp28 contains a 16 aa acidic cluster that is required for pp28 trafficking to the assembly compartment (AC) and the virus assembly. However, functional signals within the acidic cluster of pp28 remain undefined. Here, we demonstrated that an acidic cluster rather than specific sorting signals was required for trafficking to the AC. Recombinant viruses with chimeric pp28 proteins expressing non-native acidic clusters exhibited delayed viral growth kinetics and decreased production of infectious virus, indicating that the native acidic cluster of pp28 was essential for wild-type virus assembly. These results suggested that the acidic cluster of pp28 has distinct functional domains required for trafficking and for efficient virus assembly. The first half (aa 44-50) of the acidic cluster was sufficient for pp28 trafficking, whereas the native acidic cluster consisting of aa 51-59 was required for the assembly of wild-type levels of infectious virus.

  17. A Tyrosine-Based Trafficking Motif of the Tegument Protein pUL71 Is Crucial for Human Cytomegalovirus Secondary Envelopment.

    Science.gov (United States)

    Dietz, Andrea N; Villinger, Clarissa; Becker, Stefan; Frick, Manfred; von Einem, Jens

    2018-01-01

    The human cytomegalovirus (HCMV) tegument protein pUL71 is required for efficient secondary envelopment and accumulates at the Golgi compartment-derived viral assembly complex (vAC) during infection. Analysis of various C-terminally truncated pUL71 proteins fused to enhanced green fluorescent protein (eGFP) identified amino acids 23 to 34 as important determinants for its Golgi complex localization. Sequence analysis and mutational verification revealed the presence of an N-terminal tyrosine-based trafficking motif (YXXΦ) in pUL71. This led us to hypothesize a requirement of the YXXΦ motif for the function of pUL71 in infection. Mutation of both the tyrosine residue and the entire YXXΦ motif resulted in an altered distribution of mutant pUL71 at the plasma membrane and in the cytoplasm during infection. Both YXXΦ mutant viruses exhibited similarly decreased focal growth and reduced virus yields in supernatants. Ultrastructurally, mutant-virus-infected cells exhibited impaired secondary envelopment manifested by accumulations of capsids undergoing an envelopment process. Additionally, clusters of capsid accumulations surrounding the vAC were observed, similar to the ultrastructural phenotype of a UL71-deficient mutant. The importance of endocytosis and thus the YXXΦ motif for targeting pUL71 to the Golgi complex was further demonstrated when clathrin-mediated endocytosis was inhibited either by coexpression of the C-terminal part of cellular AP180 (AP180-C) or by treatment with methyl-β-cyclodextrin. Both conditions resulted in a plasma membrane accumulation of pUL71. Altogether, these data reveal the presence of a functional N-terminal endocytosis motif that is an important determinant for intracellular localization of pUL71 and that is furthermore required for the function of pUL71 during secondary envelopment of HCMV capsids at the vAC. IMPORTANCE Human cytomegalovirus (HCMV) is the leading cause of birth defects among congenital virus infections and can

  18. Epstein-Barr virus large tegument protein BPLF1 contributes to innate immune evasion through interference with toll-like receptor signaling.

    Directory of Open Access Journals (Sweden)

    Michiel van Gent

    2014-02-01

    Full Text Available Viral infection triggers an early host response through activation of pattern recognition receptors, including Toll-like receptors (TLR. TLR signaling cascades induce production of type I interferons and proinflammatory cytokines involved in establishing an anti-viral state as well as in orchestrating ensuing adaptive immunity. To allow infection, replication, and persistence, (herpesviruses employ ingenious strategies to evade host immunity. The human gamma-herpesvirus Epstein-Barr virus (EBV is a large, enveloped DNA virus persistently carried by more than 90% of adults worldwide. It is the causative agent of infectious mononucleosis and is associated with several malignant tumors. EBV activates TLRs, including TLR2, TLR3, and TLR9. Interestingly, both the expression of and signaling by TLRs is attenuated during productive EBV infection. Ubiquitination plays an important role in regulating TLR signaling and is controlled by ubiquitin ligases and deubiquitinases (DUBs. The EBV genome encodes three proteins reported to exert in vitro deubiquitinase activity. Using active site-directed probes, we show that one of these putative DUBs, the conserved herpesvirus large tegument protein BPLF1, acts as a functional DUB in EBV-producing B cells. The BPLF1 enzyme is expressed during the late phase of lytic EBV infection and is incorporated into viral particles. The N-terminal part of the large BPLF1 protein contains the catalytic site for DUB activity and suppresses TLR-mediated activation of NF-κB at, or downstream of, the TRAF6 signaling intermediate. A catalytically inactive mutant of this EBV protein did not reduce NF-κB activation, indicating that DUB activity is essential for attenuating TLR signal transduction. Our combined results show that EBV employs deubiquitination of signaling intermediates in the TLR cascade as a mechanism to counteract innate anti-viral immunity of infected hosts.

  19. Retirada do tegumento e da extração dos pigmentos na qualidade protéica do feijão-preto Effect of removal of tegument and extraction of pigments on protein quality of black beans

    Directory of Open Access Journals (Sweden)

    Ana Cristina Nascimento Chiaradia

    1999-08-01

    Full Text Available O feijão é cultivado e consumido mundialmente e é a leguminosa mais consumida na América Latina. O baixo valor biológico das proteínas do feijão pode ser resultante, dentre outros, da presença de fatores antinutricionais presentes no tegumento do grão. Este trabalho visou avaliar, através de ensaios biológicos, o efeito da retirada do tegumento e das antocianinas e outros polifenóis do feijão-preto, na qualidade de suas proteínas. Foram analisados o quociente de eficiência protéica, o quociente de eficiência líquida protéica e a utilização líquida da proteína, e digestibilidade. Os resultados obtidos mostraram que a eliminação parcial do tegumento dos grãos cozidos e secagem em estufa diminuíram significativamente o valor do quociente de eficiência protéica comparativamente ao do grão integral cozido e secado nas mesmas condições. A extração, com etanol acidificado, de 27,5% dos polifenóis não influiu significativamente no valor protéico do feijão-preto. Concluiu-se que a retirada do tegumento e a extração parcial de polifenóis do feijão não elevaram a sua qualidade protéica.Bean seeds (Phaseolus vulgaris, L. are cultivated and consumed all over the world and are a staple food in most countries of South America. The low biological value of bean proteins may be due to, among others, the effect of antinutritional factors on the tegument. The objective of this work was to evaluate, through biological assays, the effect of removal of black bean tegument, anthocyanins and other polyphenols on protein quality. The protein efficiency ratio, the protein liquid efficiency ratio, the protein liquid utilization and the digestibility were analyzed. The results indicated that removal of the tegument of cooked beans reduced their protein value (PER when compared to whole cooked beans. The ethanolic extraction of 27.5% of polyphenols did not increase the protein quality of beans. It was concluded that removal of the

  20. Classificação de lotes comerciais de feijão por meio da claridade do tegumento dos grãos Classify the common bean commerciais lots by the clarity of the grain tegument

    Directory of Open Access Journals (Sweden)

    Nerinéia Dalfollo Ribeiro

    2008-10-01

    Full Text Available A avaliação da claridade do tegumento dos grãos de feijão (valor de "L" é uma medida recente e ainda pouco utilizada no melhoramento. Por isso, o objetivo deste trabalho foi determinar se é possível classificar genótipos de feijão pelo valor de "L", dentro de mesmo grupo comercial, e avaliar o número de repetições necessárias. Dezenove ensaios foram conduzidos em Santa Maria, RS, durante os anos de 1998/1999 a 2005/2006, com número variável de genótipos. A coloração do tegumento dos grãos foi quantificada com colorímetro em grãos com 13% de umidade, imediatamente após a colheita. Considerando que a claridade dos grãos é associada ao valor comercial da cultivar, apenas os valores de "L" foram submetidos às análises. Valores entre 0,50 e 3,33 para o valor de "L" foram significativamente diferentes pelo teste de Tukey. A claridade do tegumento dos grãos pode ser utilizada para classificar lotes de feijão de um mesmo grupo comercial. Duas repetições são suficientes para a avaliação da coloração do tegumento dos grãos em feijão, com 90% de exatidão no prognóstico de seu valor real, e com alta precisão experimental.The estimation of the common bean grain clarity ('L' value is a new measure and it is not much used in the breeding programs. The objective of this research was to determine the possibility to classifying common bean genotypes for the 'L' value, within the same commercial group and to evaluate how many repetitions are necessary. Nineteen experiments were conducted in Santa Maria, in Rio Grande do Sul State, Brazil, from 1998/1999 to 2005/2006 years. The grain tegument colors were determined with 13% moisture. The clarity of grain tegument is associated with the commercial value of common bean cultivar. Values between 0.50 and 3.33 were significative different by Tukey test. The clarity of grains tegument ('L' value can be used to classify common bean lots of same commercial group. Two repetitions are

  1. A proteomic perspective of inbuilt viral protein regulation: pUL46 tegument protein is targeted for degradation by ICP0 during herpes simplex virus type 1 infection.

    Science.gov (United States)

    Lin, Aaron E; Greco, Todd M; Döhner, Katinka; Sodeik, Beate; Cristea, Ileana M

    2013-11-01

    Much like the host cells they infect, viruses must also regulate their life cycles. Herpes simples virus type 1 (HSV-1), a prominent human pathogen, uses a promoter-rich genome in conjunction with multiple viral trans-activating factors. Following entry into host cells, the virion-associated outer tegument proteins pUL46 and pUL47 act to increase expression of viral immediate-early (α) genes, thereby helping initiate the infection life cycle. Because pUL46 has gone largely unstudied, we employed a hybrid mass spectrometry-based approach to determine how pUL46 exerts its functions during early stages of infection. For a spatio-temporal characterization of pUL46, time-lapse microscopy was performed in live cells to define its dynamic localization from 2 to 24 h postinfection. Next, pUL46-containing protein complexes were immunoaffinity purified during infection of human fibroblasts and analyzed by mass spectrometry to investigate virus-virus and virus-host interactions, as well as post-translational modifications. We demonstrated that pUL46 is heavily phosphorylated in at least 23 sites. One phosphorylation site matched the consensus 14-3-3 phospho-binding motif, consistent with our identification of 14-3-3 proteins and host and viral kinases as specific pUL46 interactions. Moreover, we determined that pUL46 specifically interacts with the viral E3 ubiquitin ligase ICP0. We demonstrated that pUL46 is partially degraded in a proteasome-mediated manner during infection, and that the catalytic activity of ICP0 is responsible for this degradation. This is the first evidence of a viral protein being targeted for degradation by another viral protein during HSV-1 infection. Together, these data indicate that pUL46 levels are tightly controlled and important for the temporal regulation of viral gene expression throughout the virus life cycle. The concept of a structural virion protein, pUL46, performing nonstructural roles is likely to reflect a theme common to many viruses

  2. A Dual-Modality Herpes Simplex Virus 2 Vaccine for Preventing Genital Herpes by Using Glycoprotein C and D Subunit Antigens To Induce Potent Antibody Responses and Adenovirus Vectors Containing Capsid and Tegument Proteins as T Cell Immunogens.

    Science.gov (United States)

    Awasthi, Sita; Mahairas, Gregory G; Shaw, Carolyn E; Huang, Meei-Li; Koelle, David M; Posavad, Christine; Corey, Lawrence; Friedman, Harvey M

    2015-08-01

    We evaluated a genital herpes prophylactic vaccine containing herpes simplex virus 2 (HSV-2) glycoproteins C (gC2) and D (gD2) to stimulate humoral immunity and UL19 (capsid protein VP5) and UL47 (tegument protein VP13/14) as T cell immunogens. The HSV-2 gC2 and gD2 proteins were expressed in baculovirus, while the UL19 and UL47 genes were expressed from replication-defective adenovirus vectors. Adenovirus vectors containing UL19 and UL47 stimulated human and murine CD4(+) and CD8(+) T cell responses. Guinea pigs were either (i) mock immunized; (ii) immunized with gC2/gD2, with CpG and alum as adjuvants; (iii) immunized with the UL19/UL47 adenovirus vectors; or (iv) immunized with the combination of gC2/gD2-CpG/alum and the UL19/UL47 adenovirus vectors. Immunization with gC2/gD2 produced potent neutralizing antibodies, while UL19 and UL47 also stimulated antibody responses. After intravaginal HSV-2 challenge, the mock and UL19/UL47 adenovirus groups developed severe acute disease, while 2/8 animals in the gC2/gD2-only group and none in the combined group developed acute disease. No animals in the gC2/gD2 or combined group developed recurrent disease; however, 5/8 animals in each group had subclinical shedding of HSV-2 DNA, on 15/168 days for the gC2/gD2 group and 13/168 days for the combined group. Lumbosacral dorsal root ganglia were positive for HSV-2 DNA and latency-associated transcripts for 5/8 animals in the gC2/gD2 group and 2/8 animals in the combined group. None of the differences comparing the gC2/gD2-only group and the combined group were statistically significant. Therefore, adding the T cell immunogens UL19 and UL47 to the gC2/gD2 vaccine did not significantly reduce genital disease and vaginal HSV-2 DNA shedding compared with the excellent protection provided by gC2/gD2 in the guinea pig model. HSV-2 infection is a common cause of genital ulcer disease and a significant public health concern. Genital herpes increases the risk of transmission and

  3. Efeitos da interação genótipo x ambiente no ciclo e na coloração do tegumento dos grãos do feijoeiro comum Genotype x environment interaction efects in cycle grain in common tegument colour and cycle in bean cultivars

    Directory of Open Access Journals (Sweden)

    Nerinéia Dalfollo Ribeiro

    2004-12-01

    Full Text Available O objetivo deste trabalho foi estudar os efeitos da interação genótipo x ambiente, visando identificar cultivares de feijão com estabilidade para a coloração do tegumento dos grãos e para o ciclo na região da Depressão Central do Rio Grande do Sul. Seis experimentos foram instalados nos anos agrícolas 2000/01, 2001/02 e 2002/03, nos cultivos de safra (semeadura em setembro-outubro e de safrinha (semeadura em janeiro-fevereiro, em área do Departamento de Fitotecnia da Universidade Federal de Santa Maria. O delineamento experimental utilizado foi o de blocos ao acaso, com três repetições, e os tratamentos consistiram de 16 cultivares de feijão. Os resultados evidenciaram que a coloração do tegumento dos grãos em cultivares de feijoeiro comum do grupo comercial carioca é influenciada pelo ambiente. As cultivares Carioca, Diamante Negro, TPS Nobre e TPS Bionobre apresentam alta previsibilidade para obter grãos de feijão de cor de tegumento adequados às exigências do mercado consumidor. Nenhuma cultivar de feijão apresentou estabilidade para ciclo.The objective this work was to assess the effects of genotype x environment interaction in order to identify bean cultivars with stability for grain tegument colour and cycle in the central depression region of Rio Grande do Sul in order to guide breeding programs. Six experiments, with 16 common bean cultivars were conducted during the 2000/01, 2001/02 and 2002/03 growing season and sowing was carried out on Set/Out (Crop 1 and on Jan/Fev (Crop 2 in experimental fields of the Plant Science Department of the Santa Maria Federal University. Complete randomized blocks with three replications as used. Results showed that colour of grain tegument in carioca beans was influenciated by the environment. 'Carioca', 'Diamante Negro', 'TPS Nobre' and 'TPS Bionobre' showed high predictability for production of bean grains with tegument colour with acceptable preference by consumers. Nothing

  4. Human Asymptomatic Epitopes Identified from the Herpes Simplex Virus Tegument Protein VP13/14 (UL47) Preferentially Recall Polyfunctional Effector Memory CD44high CD62Llow CD8+ TEM Cells and Protect Humanized HLA-A*02:01 Transgenic Mice against Ocular Herpesvirus Infection.

    Science.gov (United States)

    Srivastava, Ruchi; Khan, Arif A; Garg, Sumit; Syed, Sabrina A; Furness, Julie N; Vahed, Hawa; Pham, Tiffany; Yu, Howard T; Nesburn, Anthony B; BenMohamed, Lbachir

    2017-01-15

    Herpes simplex virus 1 (HSV-1) infection is widespread among humans. The HSV-1 virion protein 13/14 (VP13/14), also known as UL47, is a tegument antigen targeted by CD8 + T cells from HSV-seropositive individuals. However, whether VP13/14-specific CD8 + T cells play a role in the natural protection seen in asymptomatic (ASYMP) individuals (individuals who have never had a clinical herpetic disease) has not been elucidated. Using predictive computer-assisted algorithms, we identified 10 potential HLA-A*02:01-restricted CD8 + T-cell epitopes from the 693-amino-acid sequence of the VP13/14 protein. Three out of 10 epitopes exhibited a high to moderate affinity of binding to soluble HLA-A*02:01 molecules. The phenotype and function of CD8 + T cells specific for each epitope were compared in HLA-A*02:01-positive ASYMP individuals and symptomatic (SYMP) individuals (individuals who have frequent clinical herpetic diseases) using determination of a combination of tetramer frequency and the levels of granzyme B, granzyme K, perforin, gamma interferon, tumor necrosis factor alpha, and interleukin-2 production and CD107 a/b cytotoxic degranulation. High frequencies of multifunctional CD8 + T cells directed against three epitopes, VP13/14 from amino acids 286 to 294 (VP13/14 286-294 ), VP13/14 from amino acids 504 to 512 (VP13/14 504-512 ), and VP13/14 from amino acids 544 to 552 (VP13/14 544-552 ), were detected in ASYMP individuals, while only low frequencies were detected in SYMP individuals. The three epitopes also predominantly recalled more CD45RA low CD44 high CCR7 low CD62L low CD8 + effector memory T cells (T EM cells) in ASYMP individuals than SYMP individuals. Moreover, immunization of HLA-A*02:01 transgenic mice with the three CD8 + T EM -cell epitopes from ASYMP individuals induced robust and polyfunctional HSV-specific CD8 + T EM cells associated with strong protective immunity against ocular herpesvirus infection and disease. Our findings outline the phenotypic

  5. HLA-A02:01-restricted epitopes identified from the herpes simplex virus tegument protein VP11/12 preferentially recall polyfunctional effector memory CD8+ T cells from seropositive asymptomatic individuals and protect humanized HLA-A*02:01 transgenic mice against ocular herpes.

    Science.gov (United States)

    Srivastava, Ruchi; Khan, Arif A; Spencer, Doran; Vahed, Hawa; Lopes, Patricia P; Thai, Nhi Thi Uyen; Wang, Christine; Pham, Thanh T; Huang, Jiawei; Scarfone, Vanessa M; Nesburn, Anthony B; Wechsler, Steven L; BenMohamed, Lbachir

    2015-03-01

    The HSV type 1 tegument virion phosphoprotein (VP) 11/12 (VP11/12) is a major Ag targeted by CD8(+) T cells from HSV-seropositive individuals. However, whether and which VP11/12 epitope-specific CD8(+) T cells play a role in the "natural" protection seen in seropositive healthy asymptomatic (ASYMP) individuals (who have never had clinical herpes disease) remain to be determined. In this study, we used multiple prediction computer-assisted algorithms to identify 10 potential HLA-A*02:01-restricted CD8(+) T cell epitopes from the 718-aa sequence of VP11/12. Three of 10 epitopes exhibited high-to-moderate binding affinity to HLA-A*02:01 molecules. In 10 sequentially studied HLA-A*02:01-positive and HSV-1-seropositive ASYMP individuals, the most frequent, robust, and polyfunctional effector CD8(+) T cell responses, as assessed by a combination of tetramer frequency, granzyme B, granzyme K, perforin, CD107(a/b) cytotoxic degranulation, IFN-γ, and multiplex cytokines assays, were predominantly directed against three epitopes: VP11/1266-74, VP11/12220-228, and VP11/12702-710. Interestingly, ASYMP individuals had a significantly higher proportion of CD45RA(low)CCR7(low)CD44(high)CD62L(low)CD27(low)CD28(low)CD8(+) effector memory CD8(+) T cells (TEMs) specific to the three epitopes, compared with symptomatic individuals (with a history of numerous episodes of recurrent ocular herpetic disease). Moreover, immunization of HLA-A*02:01 transgenic mice with the three ASYMP CD8(+) TEM cell epitopes induced robust and polyfunctional epitope-specific CD8(+) TEM cells that were associated with a strong protective immunity against ocular herpes infection and disease. Our findings outline phenotypic and functional features of protective HSV-specific CD8(+) T cells that should guide the development of an effective T cell-based herpes vaccine. Copyright © 2015 by The American Association of Immunologists, Inc.

  6. Nuclear body formation and PML body remodeling by the human cytomegalovirus protein UL35

    International Nuclear Information System (INIS)

    Salsman, Jayme; Wang Xueqi; Frappier, Lori

    2011-01-01

    The human cytomegalovirus (HCMV) UL35 gene encodes two proteins, UL35 and UL35a. Expression of UL35 in transfected cells results in the formation of UL35 nuclear bodies that associate with promyelocytic leukemia (PML) protein. PML forms the basis for PML nuclear bodies that are important for suppressing viral lytic gene expression. Given the important relationship between PML and viral infection, we have further investigated the association of UL35 with PML bodies. We demonstrate that UL35 bodies form independently of PML and subsequently recruit PML, Sp100 and Daxx. In contrast, UL35a did not form bodies; however, it could bind UL35 and inhibit the formation of UL35 bodies. The HCMV tegument protein pp71 promoted the formation of UL35 bodies and the cytoplasmic localization of UL35a. Similarly, UL35a shifted pp71 to the cytoplasm. These results indicate that the interplay between UL35, UL35a and pp71 affects their subcellular localization and likely their functions throughout infection.

  7. Neurothekeoma palpebrae in association with multiple superficial angiomyxomas: Tegumental Angiomyxoma- Neurothekeoma syndrome (TAN syndrome

    Directory of Open Access Journals (Sweden)

    Tan Aik Kah

    2011-09-01

    Full Text Available We report a case of 10-year-old Indian girl with history of multiple superficial angiomyxoma, presented with three months history of painless right upper lid swelling. There were no visual dysfunctions. Previously, the patient had multiple superficial angiomyxoma (left pinna, left upper cheek, left upper limb, chest, right axilla, hard palate and epidermal cyst (chin. The histopathological specimens were negative to S-100 protein antibody. Systemic review and family history was unremarkable. Excision biopsy and upper lid reconstruction were performed. Intraoperatively the tumor was multilobulated, firm, well encapsulated and did not invade the underlying tarsal plate. Histopathological features of the upperlid tumor were consistent with nerves sheath myxoma (neurothekeoma. To the best of the authors’ knowledge, this is the first reported case of neurothekeoma in association with multiple superficial angiomyxoma.

  8. Neurothekeoma palpebrae in association with multiple superficial angiomyxomas: Tegumental Angiomyxoma-Neurothekeoma syndrome (TAN syndrome).

    Science.gov (United States)

    Kah, Tan Aik; Yong, Ku Chui; Annuar, Faridah Hanom

    2011-07-01

    We report a case of 10-year-old Indian girl with history of multiple superficial angiomyxoma, presented with three months history of painless right upper lid swelling. There were no visual dysfunctions. Previously, the patient had multiple superficial angiomyxoma (left pinna, left upper cheek, left upper limb, chest, right axilla, hard palate) and epidermal cyst (chin). The histopathological specimens were negative to S-100 protein antibody. Systemic review and family history was unremarkable. Excision biopsy and upper lid reconstruction were performed. Intraoperatively the tumor was multilobulated, firm, well encapsulated and did not invade the underlying tarsal plate. Histopathological features of the upperlid tumor were consistent with nerves sheath myxoma (neurothekeoma). To the best of the authors' knowledge, this is the first reported case of neurothekeoma in association with multiple superficial angiomyxoma.

  9. Characterization of nuclear localization and export signals of the major tegument protein VP8 of bovine herpesvirus-1

    International Nuclear Information System (INIS)

    Zheng Chunfu; Brownlie, Robert; Babiuk, Lorne A.; Hurk, Sylvia van Drunen Littel-van den

    2004-01-01

    Bovine herpesvirus-1 (BHV-1) VP8 is found in the nucleus immediately after infection. Transient expression of VP8 fused to yellow fluorescent protein (YFP) in COS-7 cells confirmed the nuclear localization of VP8 in the absence of other viral proteins. VP8 has four putative nuclear localization signals (NLS). Deletion of pat4 ( 51 RRPR 54 ) or pat7 ( 48 PRVRRPR 54 ) NLS2 abrogated nuclear accumulation, whereas deletion of 48 PRV 50 did not, so pat4 NLS2 is critical for nuclear localization of VP8. Furthermore, NLS1 ( 11 RRPRR 15 ), pat4 NLS2, and pat7 NLS2 were all capable of transporting the majority of YFP to the nucleus. Finally, a 12-amino-acid peptide with the sequence RRPRRPRVRRPR directed all of YFP into the nucleus, suggesting that reiteration of the RRPR motif makes the nuclear localization more efficient. Heterokaryon assays demonstrated that VP8 is also capable of shuttling between the nucleus and cytoplasm of the cell. Deletion mutant analysis revealed that this property is attributed to a leucine-rich nuclear export sequence (NES) consisting of amino acids 485 LSAYLTLFVAL 495 . This leucine-rich NES caused transport of YFP to the cytoplasm. These results demonstrate that VP8 shuttles between the nucleus and cytoplasm

  10. Evaluation of conventional therapeutic methods versus maggot therapy in the evolution of healing of tegumental injuries in Wistar rats with and without diabetes mellitus.

    Science.gov (United States)

    Masiero, Franciéle Souza; Thyssen, Patricia Jacqueline

    2016-06-01

    Larval therapy consists on the application of sterilized carrion flies larvae, reared in laboratory, on acute, chronic, and/or infected wounds in order to promote healing. Conventional methods for treating injuries include mechanical debridement or silver-based dressings; however, they are not always effective for wound healing. Larval therapy is a feasible and safe treatment for therapeutic application and, in many cases, the only and the most recommended alternative for difficult healing injuries. Thus, this study aimed to evaluate the competence of Cochliomyia macellaria F. (Diptera: Calliphoridae) as a suitable species for therapeutic application and evaluate time and effectiveness of the types of treatments most commonly used to treat integumental injuries. C. macellaria eggs were obtained from colonies established in laboratory and sterilized prior to application. Twenty-five larvae were applied for each centimeter squared of lesion. Lesions were induced in 24 Wistar rats; type 1 diabetes mellitus was induced in 12 of them. Animals were divided in four groups with three individuals each, being denominated: larval therapy, larval therapy associated with foam dressing with silver release, mechanical debridement with foam dressing silver and control group, without treatment. All treatments were applied once and held for 24 h. Medical application of larvae was found to be safe, as only dead tissue was removed, and efficient to accelerate healing process when compared to other treatments.

  11. Release of Glycoprotein (GP1 from the Tegumental Surface of Taenia solium by Phospholipase C from Clostridium perfringens Suggests a Novel Protein-Anchor to Membranes

    Directory of Open Access Journals (Sweden)

    Abraham Landa

    2010-01-01

    Full Text Available In order to explore how molecules are linked to the membrane surface in larval Taenia solium, whole cysticerci were incubated in the presence of phospholipase C from Clostridium perfringens (PLC. Released material was collected and analyzed in polyacrylamide gels with sodium dodecyl sulfate. Two major bands with apparent molecular weights of 180 and 43 kDa were observed. Western blot of released material and localization assays in cysticerci tissue sections using antibodies against five known surface glycoproteins of T. solium cysticerci indicated that only one, previously called GP1, was released. Similar localization studies using the lectins wheat-germ-agglutinin and Concanavalin A showed that N-acetyl-D-glucosamine, N-acetylneuraminic, sialic acid, αmethyl-D-mannoside, D-manose/glucose, and N-acetyl-D-glucosamine residues are abundantly present on the surface. On the other hand, we find that treatment with PLC releases molecules from the surface; they do not reveal Cross Reacting Determinant (CRD, suggesting a novel anchor to the membrane for the glycoprotein GP1.

  12. Ultrastructure of the proglottid tegument (neodermis) of the cestode Echinophallus wageneri (Pseudophyllidea: Echinophallidae), a parasite of the bathypelagic fish Centrolophus niger

    Czech Academy of Sciences Publication Activity Database

    Poddubnaya, L. G.; Scholz, Tomáš; Kuchta, Roman; Levron, Céline; Bruňanská, Magdaléna

    2007-01-01

    Roč. 101, č. 2 (2007), s. 373-383 ISSN 0932-0113 R&D Projects: GA ČR GA206/03/1317; GA ČR GD524/03/H133; GA ČR GA524/04/0342; GA ČR GP524/07/P039; GA MŠk LC522 Grant - others:Vedecká grantová agentúra Ministerstva školstva Slovenskej republiky a Slovenskej akadémie vied(SK) VEGA2/4177/04 Institutional research plan: CEZ:AV0Z60220518 Keywords : ultrastructure * microtriches * tapeworms Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 1.512, year: 2007

  13. Contribution to the study of external decontamination procedure. Experiments on a new product in the case of radioactive contamination of the teguments

    International Nuclear Information System (INIS)

    Toulet, J.; Tabernat, J.

    1959-01-01

    General principles of external decontamination with elements of practical organisation in a centre of nuclear studies. Reprint of a paper published in 'Archives des Maladies Professionnelles', Tome 20, n. 3, 1959, p. 272-282

  14. Human cytomegalovirus tegument protein pp65 is detected in all intra- and extra-axial brain tumours independent of the tumour type or grade.

    Directory of Open Access Journals (Sweden)

    Sylwia Libard

    Full Text Available Human cytomegalovirus (HCMV has been indicated being a significant oncomodulator. Recent reports have suggested that an antiviral treatment alters the outcome of a glioblastoma. We analysed the performance of commercial HCMV-antibodies applying the immunohistochemical (IHC methods on brain sample obtained from a subject with a verified HCMV infection, on samples obtained from 14 control subjects, and on a tissue microarray block containing cores of various brain tumours. Based on these trials, we selected the best performing antibody and analysed a cohort of 417 extra- and intra-axial brain tumours such as gliomas, medulloblastomas, primary diffuse large B-cell lymphomas, and meningiomas. HCMV protein pp65 immunoreactivity was observed in all types of tumours analysed, and the IHC expression did not depend on the patient's age, gender, tumour type, or grade. The labelling pattern observed in the tumours differed from the labelling pattern observed in the tissue with an active HCMV infection. The HCMV protein was expressed in up to 90% of all the tumours investigated. Our results are in accordance with previous reports regarding the HCMV protein expression in glioblastomas and medulloblastomas. In addition, the HCMV protein expression was seen in primary brain lymphomas, low-grade gliomas, and in meningiomas. Our results indicate that the HCMV protein pp65 expression is common in intra- and extra-axial brain tumours. Thus, the assessment of the HCMV expression in tumours of various origins and pathologically altered tissue in conditions such as inflammation, infection, and even degeneration should certainly be facilitated.

  15. Evaluasi Penerapan PP No 71/2010 Tentang Standar Akuntansi Pemrintahan Berbasis Akrual: Studi Kasus di Pemda Nganjuk

    Directory of Open Access Journals (Sweden)

    Dahlia Firdaus

    2016-03-01

    Full Text Available The purpose of this research was to determine the suitability of the accrual basis implementation on Governmental Accounting Standards in Peraturan Pemerintah (PP No. 71/2010 at Nganjuk Regency’s Financial Statements year 2013. This research related with Dinas Pendapatan, Pengelolaan Keuangan dan Aset Daerah (Department of Revenue, Finance and Asset Management. Research proves that Nganjuk Regency’s Financial Statements not yet fully implementing the PP 71/2010. Nganjuk Regency’s financial statements still apply Enclosure II of PP 71/2010 that is cash toward accrual basis. This is due to the application of PP 71/2010 accrual basis (Enclosure I of PP 71/010 are required to be implemented in fiscal year 2015.

  16. Resonance – Journal of Science Education | Indian Academy of ...

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 7; Issue 12. Our Shrinking Ozone Layer - Global Perspectives and Remedial Measures. Shashi K Pathak Nigel J Mason. General Article Volume 7 Issue 12 December 2002 pp 71-80 ...

  17. Identification of structural protein-protein interactions of herpes simplex virus type 1.

    Science.gov (United States)

    Lee, Jin H; Vittone, Valerio; Diefenbach, Eve; Cunningham, Anthony L; Diefenbach, Russell J

    2008-09-01

    In this study we have defined protein-protein interactions between the structural proteins of herpes simplex virus type 1 (HSV-1) using a LexA yeast two-hybrid system. The majority of the capsid, tegument and envelope proteins of HSV-1 were screened in a matrix approach. A total of 40 binary interactions were detected including 9 out of 10 previously identified tegument-tegument interactions (Vittone, V., Diefenbach, E., Triffett, D., Douglas, M.W., Cunningham, A.L., and Diefenbach, R.J., 2005. Determination of interactions between tegument proteins of herpes simplex virus type 1. J. Virol. 79, 9566-9571). A total of 12 interactions involving the capsid protein pUL35 (VP26) and 11 interactions involving the tegument protein pUL46 (VP11/12) were identified. The most significant novel interactions detected in this study, which are likely to play a role in viral assembly, include pUL35-pUL37 (capsid-tegument), pUL46-pUL37 (tegument-tegument) and pUL49 (VP22)-pUS9 (tegument-envelope). This information will provide further insights into the pathways of HSV-1 assembly and the identified interactions are potential targets for new antiviral drugs.

  18. Dicty_cDB: Contig-U11810-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available otein; Short=MT... 78 8e-13 S55598( S55598 ) tegument protein 03 - equine herpesv...( S55669 ) tegument protein 75 - equine herpesvirus 2 &U208... 61 1e-07 AE009950_198( AE009950 |pid:none) Py

  19. AcEST: BP911450 [AcEST

    Lifescience Database Archive (English)

    Full Text Available EGU_VZVD Large tegument protein OS=Varicella-zoster virus (strain Dumas) Align length 93 Score (bit) 31.2 E-...8.3 >sp|P09278|TEGU_VZVD Large tegument protein OS=Varicella-zoster virus (strain Dumas) GN=22 PE=3 SV=1 Len

  20. NCBI nr-aa BLAST: CBRC-AGAM-05-0030 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-AGAM-05-0030 ref|NP_851896.1| large tegument protein [Cercopithecine herpesvir...us 1] gb|AAP41454.1| very large tegument protein [Cercopithecine herpesvirus 1] NP_851896.1 3e-07 33% ...

  1. NCBI nr-aa BLAST: CBRC-EEUR-01-1253 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-EEUR-01-1253 ref|NP_851896.1| large tegument protein [Cercopithecine herpesvir...us 1] gb|AAP41454.1| very large tegument protein [Cercopithecine herpesvirus 1] NP_851896.1 0.010 28% ...

  2. NCBI nr-aa BLAST: CBRC-PVAM-01-0933 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-PVAM-01-0933 ref|YP_164479.1| large tegument protein [Cercopithecine herpesvir...us 2] gb|AAU88102.1| very large tegument protein [Cercopithecine herpesvirus 2] YP_164479.1 0.009 24% ...

  3. NCBI nr-aa BLAST: CBRC-MDOM-02-0463 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-02-0463 ref|NP_954911.1| large tegument protein [Bovine herpesvirus 5] gb...|AAR86125.1| UL36 very large tegument protein [Bovine herpesvirus 5] NP_954911.1 8e-11 34% ...

  4. Gödel's Proof

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 12; Issue 5. Gödel's Proof - Incompleteness Theorems. S M Srivastava. General Article Volume 12 Issue 5 May 2007 pp 71-84. Fulltext. Click here to view fulltext PDF. Permanent link: https://www.ias.ac.in/article/fulltext/reso/012/05/0071-0084. Keywords.

  5. Can you see Air?

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 12; Issue 6. Can you see Air? Andal Narayanan Joseph Samuel Supurna Sinha. Classroom Volume 12 Issue 6 June 2007 pp 71-75. Fulltext. Click here to view fulltext PDF. Permanent link: https://www.ias.ac.in/article/fulltext/reso/012/06/0071-0075 ...

  6. Resonance – Journal of Science Education | Indian Academy of ...

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 8; Issue 11. Origami, Modular Packing and the Soma Puzzle. Subramania Ranganathan. General Article Volume 8 Issue 11 November 2003 pp 71-77. Fulltext. Click here to view fulltext PDF. Permanent link:

  7. Evolution of a Test Item

    Science.gov (United States)

    Spaan, Mary

    2007-01-01

    This article follows the development of test items (see "Language Assessment Quarterly", Volume 3 Issue 1, pp. 71-79 for the article "Test and Item Specifications Development"), beginning with a review of test and item specifications, then proceeding to writing and editing of items, pretesting and analysis, and finally selection of an item for a…

  8. Journal of Biosciences | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    Replica-moulded polydimethylsiloxane culture vessel lids attenuate osmotic drift in long-term cell cultures · Axel Blau Tanja Neumann Christiane Ziegler Fabio Benfenati ... otherwise conventional cell culture conditions. pp 71-83 Articles. Molecular evolution of the E8 promoter in tomato and some of its relative wild species.

  9. Sadhana | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    Home; Journals; Sadhana; Volume 33; Issue 2. Issue front cover thumbnail. Volume 33, Issue 2. April 2008, pages 69-174. pp 69-70. Editorial · N Mukunda R N Iyengar · More Details Fulltext PDF. pp 71-82. Emission control with route optimization in solid waste collection process: A case study · Omer Apaydin M Talha ...

  10. C Sacerdoti Coen

    Indian Academy of Sciences (India)

    Home; Journals; Sadhana. C Sacerdoti Coen. Articles written in Sadhana. Volume 34 Issue 1 February 2009 pp 71-144. A compact kernel for the calculus of inductive constructions · A Asperti W Ricciotti C Sacerdoti Coen E Tassi · More Details Abstract Fulltext PDF. The paper describes the new kernel for the Calculus of ...

  11. Journal of Chemical Sciences | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    Home; Journals; Journal of Chemical Sciences; Volume 117; Issue 2. Volume 117, Issue 2. March 2005, pages 71-201. Recent Advances in Bio-Inorganic Chemistry. pp 71-71. Foreword · T K Chandrashekar · More Details Fulltext PDF. pp 73-84. Meso-functionalized octamethoxyporphyrins: A new class of nonasubstituted ...

  12. Resonance – Journal of Science Education | News

    Indian Academy of Sciences (India)

    A W Joshi Alok Kumar · More Details Fulltext PDF. pp 26-31 General Article. A Promising Approach to the Twin Prime Problem ... More Details Fulltext PDF. pp 71-80 Classroom. Brownian Motion: Theory and Experiment A Simple Classroom Measurement of the Diffusion Coefficient · Kasturi Basu Kopijol Baishya.

  13. Download this PDF file

    African Journals Online (AJOL)

    ADOWIE PERE

    chromatography (GC). The results showed that the double salts were able to alter the ... Keywords: Diesel, double salt, Hydrocarbons, Gas Chromatography. The largest source of hydrocarbons found in nature is crude oil. .... Toxicology: In theory and practice. Jeso. International, Owerri, pp71-90. Wellinton, B F, Asmus, A F ...

  14. Microfloral assemblage, age and paleoenvironment of the Upper ...

    African Journals Online (AJOL)

    Microfloral assemblage, age and paleoenvironment of the Upper Cretaceous Patti Formation, southeastern Bida Basin, Nigeria. OJ Ojo, SO Akande. Abstract. No Abstract. Journal of Mining and Geology Vol. 44 (1) 2008: pp. 71-82. Full Text: EMAIL FULL TEXT EMAIL FULL TEXT · DOWNLOAD FULL TEXT DOWNLOAD ...

  15. Hydraulic and hydrochemical characteristics of the phreatic ...

    African Journals Online (AJOL)

    Hydraulic and hydrochemical characteristics of the phreatic basement aquifers in parts of southwestern Nigeria. OA Idowu, O Martins, AM Gbadebo. Abstract. No Abstract. Journal of Mining and Geology Vol. 43 (1) 2007: pp. 71-78. Full Text: EMAIL FULL TEXT EMAIL FULL TEXT · DOWNLOAD FULL TEXT DOWNLOAD ...

  16. Resonance – Journal of Science Education | Indian Academy of ...

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 6; Issue 2. On How to Arrange Infinitely many Marriages! B Sury. Classroom Volume 6 Issue 2 February 2001 pp 71-73. Fulltext. Click here to view fulltext PDF. Permanent link: https://www.ias.ac.in/article/fulltext/reso/006/02/0071-0073. Author Affiliations.

  17. Crowns and Crypts

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 6; Issue 6. Crowns and Crypts - A Fascinating Group of Multidentate Macrocyclic Ligands. Debasis Bandyopadhyay. General Article Volume 6 Issue 6 June 2001 pp 71-79. Fulltext. Click here to view fulltext PDF. Permanent link:

  18. Journal of Biosciences | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    Home; Journals; Journal of Biosciences. M T Tanuja. Articles written in Journal of Biosciences. Volume 26 Issue 1 March 2001 pp 71-76 Articles. Incipient sexual isolation in the nasuta-albomicans complex of Drosophila: No-choice experiments · M T Tanuja N B Ramachandra H A Ranganath · More Details Abstract Fulltext ...

  19. Sadhana | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    Home; Journals; Sadhana. M Talha Gonullu. Articles written in Sadhana. Volume 33 Issue 2 April 2008 pp 71-82. Emission control with route optimization in solid waste collection process: A case study · Omer Apaydin M Talha Gonullu · More Details Abstract Fulltext PDF. Solid waste collection processes are usually carried ...

  20. Moselio Schaechter

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education. Moselio Schaechter. Articles written in Resonance – Journal of Science Education. Volume 12 Issue 2 February 2007 pp 71-82 Reflections. Integrative Microbiology - The Third Golden Age · Moselio Schaechter · More Details Fulltext PDF. Volume 13 Issue 2 ...

  1. Resonance – Journal of Science Education | News

    Indian Academy of Sciences (India)

    Natural Nano-Machines - Discussion on Methods, Materials and Mechanisms · Debashish Chowdhury · More Details Fulltext PDF. pp 59-70 General Article. Gödel's Proof - An Introduction to Mathematical Logic · S M Srivastava · More Details Fulltext PDF. pp 71-82 Reflections. Integrative Microbiology - The Third Golden ...

  2. Resonance – Journal of Science Education | Indian Academy of ...

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education. T Krishnan. Articles written in Resonance – Journal of Science Education. Volume 2 Issue 9 September 1997 pp 32-37 General Article. Fisher's Contributions to Statistics · T Krishnan · More Details Fulltext PDF. Volume 4 Issue 3 March 1999 pp 71-73 Classroom.

  3. F F Sun

    Indian Academy of Sciences (India)

    Home; Journals; Bulletin of Materials Science. F F Sun. Articles written in Bulletin of Materials Science. Volume 37 Issue 1 February 2014 pp 71-76. Study of electroless copper plating on ABS resin surface modified by heterocyclic organosilane self-assembled film · H N Zhang J Wang F F Sun D Liu H Y Wang F Wang.

  4. Fermat's Last Theorem A Theorem at Last!

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 1; Issue 1. Fermat's Last Theorem A Theorem at Last! C S Yogananda. General Article Volume 1 Issue 1 January 1996 pp 71-79. Fulltext. Click here to view fulltext PDF. Permanent link: https://www.ias.ac.in/article/fulltext/reso/001/01/0071-0079 ...

  5. Properties of virion transactivator proteins encoded by primate cytomegaloviruses

    Directory of Open Access Journals (Sweden)

    Barry Peter A

    2009-05-01

    Full Text Available Abstract Background Human cytomegalovirus (HCMV is a betaherpesvirus that causes severe disease in situations where the immune system is immature or compromised. HCMV immediate early (IE gene expression is stimulated by the virion phosphoprotein pp71, encoded by open reading frame (ORF UL82, and this transactivation activity is important for the efficient initiation of viral replication. It is currently recognized that pp71 acts to overcome cellular intrinsic defences that otherwise block viral IE gene expression, and that interactions of pp71 with the cell proteins Daxx and ATRX are important for this function. A further property of pp71 is the ability to enable prolonged gene expression from quiescent herpes simplex virus type 1 (HSV-1 genomes. Non-human primate cytomegaloviruses encode homologs of pp71, but there is currently no published information that addresses their effects on gene expression and modes of action. Results The UL82 homolog encoded by simian cytomegalovirus (SCMV, strain Colburn, was identified and cloned. This ORF, named S82, was cloned into an HSV-1 vector, as were those from baboon, rhesus monkey and chimpanzee cytomegaloviruses. The use of an HSV-1 vector enabled expression of the UL82 homologs in a range of cell types, and permitted investigation of their abilities to direct prolonged gene expression from quiescent genomes. The results show that all UL82 homologs activate gene expression, and that neither host cell type nor promoter target sequence has major effects on these activities. Surprisingly, the UL82 proteins specified by non-human primate cytomegaloviruses, unlike pp71, did not direct long term expression from quiescent HSV-1 genomes. In addition, significant differences were observed in the intranuclear localization of the UL82 homologs, and in their effects on Daxx. Strikingly, S82 mediated the release of Daxx from nuclear domain 10 substructures much more rapidly than pp71 or the other proteins tested. All

  6. Effects of the humic acid extracted from vermicompost on the ...

    African Journals Online (AJOL)

    Felipe Andrade

    2015-05-06

    May 6, 2015 ... Organic matter for soils, waters and sediments have humic substances as their ... the essays (Kononova, 1982; Santos and Camargo,. 1999; Muscolo et al., .... maturity, which influences the permeability of the tegument and ...

  7. Penerapan Sistem Akuntansi Instansi Berbasis Akrual sebagai Implementasi PP No. 71 Tahun 2010 pada Politeknik Negeri Pontianak

    OpenAIRE

    Theresia Siwi Kartikawati 3), Sari Zawitri 1) Ninik Kurniasih 2)

    2015-01-01

    The government regulation (PP) number 71 year 2010 is a new regulation standard of accounting in the public sector. The new regulation standard that inspiring the writers to the research title: Penerapan Sistem Akuntansi Instansi Berbasis Akrual sebagai Implementasi PP nomor 71 tahun 2010 pada Politeknik Negeri Pontianak. The Problems are formulated as (1) The changes of institutional accounting system (SAI) PP 24 year 2005 into PP 71 year 2010 or cash toward accrual into accrual basis? (2) H...

  8. The Shock and Vibration Digest. Volume 16, Number 8

    Science.gov (United States)

    1984-08-01

    Amplitude Free Vibrations of a Square Plate of Variable Thickne- S.K. Chaudhuri 0 Acharya B.N. Seal College, Cooch - Behar , W. Bengal, 841716 India, J...Dimen- discs having variable thickness and density along their sional Stressed State of the Blades of Gas- radius [194]. Calculation of critical rotating...34 Ph.D. Thesis, Turbine Blade Vibrations," Problemy Proch- Ohio State Univ., DA 8305407 (1982). nosti, 156 (6), pp 71-74 (June 1982) (In Rus- .4 sian

  9. Deterioration of Early Holocene coral reef due to sea level rise along west coast of India: Benthic foraminiferal testimony

    Digital Repository Service at National Institute of Oceanography (India)

    Mazumder, A.; Nigam, R.; Henriques, P.J.

    and constituting the main components of the carbonate beach sands: Examples from Ryukyu archipelago, in: Proceeding FORAMS’98 – International Symposium on Foraminifera. Monterrey, Mexico. Kaminski, M.A., Aksu, A., Box, M., Hiscott, R.N., Filipescu, S., Al..., B., 1991. Climatic trends and variability, in: Jager, J., Ferguson, H.L. (Eds.), Climate change, Science, Impact and Policy. Proc. II World Climate Conference, pp. 71-82. Daniel, A., 1972. Marine intertidal barnacles in the Indian Ocean. Proceeding...

  10. Remedial Investigation Addendum Report Data Item A009. Volume 1: Report Test

    Science.gov (United States)

    1993-12-01

    placed over the vegetation. Once the square had settled, phytomacrofauna were collected from submerged macrophytes and the water column with D-frame...with aquatic macrophytes , including sweet water lily (Nymphaea odorata) and water shield (Brasenia schreberi). Submerged macrophytes (primarily water...Miller, G.J., 1984. Chemistry and Ecotoxicology of Pollution. John Wiley & Sons, New York, pp. 71-85. ABB Environmental Services, Inc. W0069310.M80 7005

  11. Applications of Strengths-Based Leadership Theory for the U.S. Army

    Science.gov (United States)

    2012-09-01

    Albert Bandura’s (1994) theory of self-efficacy also helps explain how optimization might take place. According to SBL theory, when individuals invest...improved productivity and satisfaction ( Bandura , 1994). In practice, consistent investment in talents by leaders and subordinates leads to the...References Bandura , A. (1994). Self-efficacy. In V. S. Ramachaudran (Ed.), Encyclopedia of human behavior (pp. 71-81). New York: Academic Press

  12. Validation of models in an imaging infrared simulation

    CSIR Research Space (South Africa)

    Willers, C

    2007-10-01

    Full Text Available threeprocessesfortransformingtheinformationbetweentheentities. Reality/ Problem Entity Conceptual Model Computerized Model Model Validation ModelVerification Model Qualification Computer Implementation Analysisand Modelling Simulationand Experimentation “Substantiationthata....C.Refsgaard ,ModellingGuidelines-terminology andguidingprinciples, AdvancesinWaterResources, Vol27,No1,January2004,?pp.71-82(12),Elsevier. et.al. [5]N.Oreskes,et.al.,Verification,Validation,andConfirmationof NumericalModelsintheEarthSciences,Science,Vol263, Number...

  13. Analisis Penerapan Standar Akuntansi Pemerintah terhadap Aset Tetap Pemerintah Daerah Kabupaten Bolaang Mongondow

    OpenAIRE

    Lauma, Enda Baisida; Morasa, Jenny; Kalangi, Lintje

    2016-01-01

    Government Accounting Standard have changed from Cash Basis is to comply with the Government Regulation number 24 2005s to Government Regulation number 71 2010s. Requirement to implemented Accrual basis SAP by the Local Government was start in the Local Government Financial Statement for the year ended on December 2015s. The District Government of Bolaang Mongondow, before implemented accrual basis PP 71/2010 always received non-WTP opinion, because of asset problems. The objective of the res...

  14. Penerapan Sistem Akuntansi Instansi Berbasis Akrual Sebagai Implementasi PP No. 71 Tahun 2010 pada Politeknik Negeri Pontianak

    Directory of Open Access Journals (Sweden)

    Sari Zawitri

    2015-04-01

    Full Text Available The government regulation (PP number 71 year 2010 is a new regulation standard of accounting in the public sector. The new regulation standard that inspiring the writers to the research title: Penerapan Sistem Akuntansi Instansi Berbasis Akrual sebagai Implementasi PP nomor 71 tahun 2010 pada Politeknik Negeri Pontianak. The Problems are formulated as (1 The changes of institutional accounting system (SAI PP 24 year 2005 into PP 71 year 2010 or cash toward accrual into accrual basis? (2 How preparation and the application transition accrual basis institutional accounting system (SAI or implementation PP 71 year 2010 of the Politeknik Negeri Pontianak? It’s concern to disclosure and measurement of Asset, Liabilities, Earned, and Expense. This the exploratory research method which is narrative and descriptive approach with the Politeknik Negeri Pontianak is an object. The results, as form of adjustment in order to change cash toward accrual into accrual basis, accrual accounting recognizes revenues by treasurer expenditure with adding income that still must be accepted in the accounting period during which the service is performed and/or adding unearned revenue. Politeknik Negeri Pontianak almost meets the six conditions that must have preparation and the application transition cash toward accrual into accrual basis of institutional accounting system as implementation PP 71 year 2010 be general conclusion of this research. Writers suggested all parties related in an accounting system institutional of Politeknik Negeri Pontianak commitment to keeping together in the process of financial reporting with accrual basis.

  15. Bitterness in Almonds1[C][OA

    Science.gov (United States)

    Sánchez-Pérez, Raquel; Jørgensen, Kirsten; Olsen, Carl Erik; Dicenta, Federico; Møller, Birger Lindberg

    2008-01-01

    Bitterness in almond (Prunus dulcis) is determined by the content of the cyanogenic diglucoside amygdalin. The ability to synthesize and degrade prunasin and amygdalin in the almond kernel was studied throughout the growth season using four different genotypes for bitterness. Liquid chromatography-mass spectrometry analyses showed a specific developmentally dependent accumulation of prunasin in the tegument of the bitter genotype. The prunasin level decreased concomitant with the initiation of amygdalin accumulation in the cotyledons of the bitter genotype. By administration of radiolabeled phenylalanine, the tegument was identified as a specific site of synthesis of prunasin in all four genotypes. A major difference between sweet and bitter genotypes was observed upon staining of thin sections of teguments and cotyledons for β-glucosidase activity using Fast Blue BB salt. In the sweet genotype, the inner epidermis in the tegument facing the nucellus was rich in cytoplasmic and vacuolar localized β-glucosidase activity, whereas in the bitter cultivar, the β-glucosidase activity in this cell layer was low. These combined data show that in the bitter genotype, prunasin synthesized in the tegument is transported into the cotyledon via the transfer cells and converted into amygdalin in the developing almond seed, whereas in the sweet genotype, amygdalin formation is prevented because the prunasin is degraded upon passage of the β-glucosidase-rich cell layer in the inner epidermis of the tegument. The prunasin turnover may offer a buffer supply of ammonia, aspartic acid, and asparagine enabling the plants to balance the supply of nitrogen to the developing cotyledons. PMID:18192442

  16. Ultrastructural and Histopathological Studies of the Digenetic ...

    African Journals Online (AJOL)

    The tegument of the cryptogonimid trematode is characterised with some markings and whitish ornamentations. The cephalic and the body region bears some compressed alae. The parasite has the distome body plan of digenetic trematode. The histopathology of the parasitized gastrointestinal tracts revealed necrosis of ...

  17. Ultrastructural alterations in adult Schistosoma mansoni caused by artemether

    Directory of Open Access Journals (Sweden)

    Shuhua Xiao

    2002-07-01

    Full Text Available Progress has been made over the last decade with the development and clinical use of artemether as an agent against major human schistosome parasites. The tegument has been identified as a key target of artemether, implying detailed studies on ultrastructural damage induced by this compound. We performed a temporal examination, employing a transmission electron microscope to assess the pattern and extent of ultrastructural alterations in adult Schistosoma mansoni harboured in mice treated with a single dose of 400 mg/kg artemether. Eight hours post-treatment, damage to the tegument and subtegumental structures was seen. Tegumental alterations reached a peak 3 days after treatment and were characterized by swelling, fusion of distal cytoplasma, focal lysis of the tegumental matrix and vacuolisation. Tubercles and sensory organelles frequently degenerated or collapsed. Typical features of subtegumental alterations, including muscle fibres, syncytium and parenchyma tissues, were focal or extensive lysis, vacuolisation and degeneration of mitochondria. Severe alterations were also observed in gut epithelial cells and vitelline cells of female worms. Our findings of artemether-induced ultrastructural alterations in adult S. mansoni confirm previous results obtained with juvenile S. mansoni and S. japonicum of different ages.

  18. Aporocotyle michaudi n. sp. (Digenea: Aporocotylidae) from the emerald rock cod, Trematomus bernacchii (Teleostei: Perciformes) in Antarctica.

    Science.gov (United States)

    Santoro, Mario; Cipriani, Paolo; Pankov, Plamen; Lawton, Scott P

    2015-10-01

    Aporocotyle michaudi n. sp. is described from the gill blood vessels of the emerald rock cod Trematomus bernacchii in the Ross Sea, Antarctica. It is distinguished from all other species of Aporocotyle by its body tegument showing single conical spines, spinous buccal capsule, and genital atrium positioned medially; all congeners described to date are characterized by clusters of tegumental spines, unspined buccal capsule and genital atrium located in the lateral part of the body. Aporocotyle michaudi n. sp. clearly differs from A. notothenia (the only other species of Aporocotyle found in a perciform fish) in its shape and arrangement of tegumental spines, buccal capsule features, location of genital atrium, body size, ratio of esophagus/body length, anterior caeca/posterior caeca ratio, number of testes, cirrus sac and ovary size and shape, and host. The new species is easily distinguished from A. argentinensis (the species that most closely resembles A. michaudi) by the shape and arrangement of tegumental spines, buccal capsule features, genital atrium location, left anterior caecum longer than right, esophagus/body length ratio, number of testes, cirrus sac size and shape, host and molecular analyses. Phylogenetic analyses of partial 28S rDNA genetic data showed that sequences representing the new species form a distinct clade with all other sequences for species of Aporocotyle and appear basal within the genus. Aporocotyle michaudi n. sp. represents the only species of genus described in Antarctica. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  19. Structural analysis of herpes simplex virus by optical super-resolution imaging

    Science.gov (United States)

    Laine, Romain F.; Albecka, Anna; van de Linde, Sebastian; Rees, Eric J.; Crump, Colin M.; Kaminski, Clemens F.

    2015-01-01

    Herpes simplex virus type-1 (HSV-1) is one of the most widespread pathogens among humans. Although the structure of HSV-1 has been extensively investigated, the precise organization of tegument and envelope proteins remains elusive. Here we use super-resolution imaging by direct stochastic optical reconstruction microscopy (dSTORM) in combination with a model-based analysis of single-molecule localization data, to determine the position of protein layers within virus particles. We resolve different protein layers within individual HSV-1 particles using multi-colour dSTORM imaging and discriminate envelope-anchored glycoproteins from tegument proteins, both in purified virions and in virions present in infected cells. Precise characterization of HSV-1 structure was achieved by particle averaging of purified viruses and model-based analysis of the radial distribution of the tegument proteins VP16, VP1/2 and pUL37, and envelope protein gD. From this data, we propose a model of the protein organization inside the tegument.

  20. Mitosis and cell death in the optic lobes of workers, queens and ...

    Indian Academy of Sciences (India)

    Prakash

    of white, pink, brown and black eyes, and the beginning of tegument ... Our purpose was to investigate cell division and death in the optic lobes (OL) of workers, queens and males during ... new ones in each caste and sex. Nevertheless, most ...

  1. 35 . REVIEW ARTICLE

    African Journals Online (AJOL)

    User

    spikes are dispersed evenly over the entire surface. The nucleocapsid is isometric, and surrounded by the tegument that consists of globular material which is frequently ... molecule of linear double stranded DNA. Total genome length is 120,000-220,000 .Molecular virological techniques have been used to study variation ...

  2. Gas hydrates

    Digital Repository Service at National Institute of Oceanography (India)

    Ramprasad, T.

    , not all of them are white like snow. Some hydrates from the deep Gulf of Mexico are richly colored in shades of yellow, orange, or even red. The ice-like masses are beautiful, and contrast with the dull gray of deep sea muds. Hydrates from the Blake... volcanoes and associated gas hydrates: Marine Geology, v. 167, p. 29-42. Milkov, A.V. and R. Sassen, 2001a, Estimate of gas hydrate resource, northwestern Gulf of Mexico continental slope: Marine Geology, v. 179, pp. 71-83. Milkov, A.V., Sassen, R...

  3. Where are the sensory organs of Nybelinia surmenicola (Trypanorhyncha)? A comparative analysis with Parachristianella sp. and other trypanorhynchean cestodes.

    Science.gov (United States)

    Biserova, Natalia M; Gordeev, Ilya I; Korneva, Janetta V

    2016-01-01

    The sensory organs in tegument of two trypanorhynchean species--Nybelinia surmenicola (plerocercoid) and adult Parachristianella sp. (Cestoda, Trypanorhyncha)--were studied with the aim of ultrastructural description and a comparative analysis. The Nybelinia surmenicola plerocercoid lacks papillae with sensory cilia on the bothria adhesive surface. We found an unciliated sensory organ within the median bothria fold. This unciliated free nerve ending contains the central electron-dense disc, three dense supporting rings, and broad root. The nerve ending locates in the basal matrix under the tegument. The tegument of N. surmenicola has a number of ultrastructural features which make it significantly different from other Trypanorhyncha: (i) the tegumental cytoplasm has a plicated constitution in a form of high apical and deep basal folds, (ii) numerous layers of the basal matrix are presented in the subtegument, and (iii) the squamiform and bristlelike microtriches N. surmenicola lack the base and the basal plate. In contrast, numerous ciliated and unciliated receptors were found in Parachristianella sp.: six types on the bothria and one type in the strobila tegument. Ultrastructural constitution of sensory organs in the form of ciliated free nerve endings as well as unciliated basal nerve endings of Parachristianella sp. has many common features inside Eucestoda. In comparison with other Trypanorhyncha, all Nybelinia species studied have less quantity of the bothrial sensory organs. This fact may reflect behavioral patterns of Nybelinia as well as phylogenetic position into Trypanorhyncha. Our observations of living animals conventionally demonstrate the ability of N. surmenicola plerocercoids to locomote in forward direction on the Petri dish surface. The participation of the bothrial microtriches in a parasite movement has been discussed.

  4. In vitro evaluation of the schistosomicidal effect of the extracts, fractions and major 3-hydroxy-2,6-dialkyl-substituted piperidine alkaloids from the flowers of Senna spectabilis (Fabaceae).

    Science.gov (United States)

    de Castro, Andreísa Teixeira; Castro, Aline Pereira; Silva, Matheus Siqueira; de Souza, Isabella Maria Monteiro; Martins-Souza, Raquel Lopes; Chagas-Paula, Daniela Aparecida; Coelho, Luiz Felipe Leomil; da Silva Bolzani, Vanderlan; Pivatto, Marcos; Viegas, Claudio; Marques, Marcos José

    2016-09-01

    In this work, we present the in vitro schistosomicidal activity evaluation of the most active dichloromethane fraction (FDm) (ED50=83.5μg/mL) and of a mixture of the major alkaloids ((-)-cassine/(-)-spectaline, C/E) (ED50=37.4μg/mL) from the flowers of Senna spectabilis against adult worms and cercariae. We also demonstrate other toxic effects including paralysis of the adult worms, inhibition of the secretory activity, tegument lesions and cercaricidal activity. In the association test of Praziquantel (PZQ)-C/E, we observed up to 80% mortality of Schistosoma mansoni in comparison to PZQ monotherapy. Due to the diversity of the toxic effects, the schistosomicidal activity of C/E is likely a result of a multitarget mechanism involving the tegument, secretory system and neuromotor action. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Antischistosomal Activity of the Terpene Nerolidol

    Directory of Open Access Journals (Sweden)

    Marcos P.N. Silva

    2014-03-01

    Full Text Available Schistosomiasis is a neglected tropical disease that affects hundreds of millions of people worldwide. Since the treatment of this disease currently relies on a single drug, praziquantel, new and safe schistosomicidal agents are urgently required. Nerolidol, a sesquiterpene present in the essential oils of several plants, is found in many foods and was approved by the U.S. Food and Drug Administration. In this study we analysed the in vitro antiparasitic effect of nerolidol on Schistosoma mansoni adult worms. Nerolidol at concentrations of 31.2 and 62.5 μM reduced the worm motor activity and caused the death of all male and female schistosomes, respectively. In addition, confocal laser scanning microscopy revealed morphological alterations on the tegument of worms such as disintegration, sloughing and erosion of the surface, and a correlation between viability and tegumental damage was observed. In conclusion, nerolidol may be a promising lead compound for the development of antischistosomal natural agents.

  6. Antigenic profile and localization of Clonorchis sinensis proteins in the course of infection

    Science.gov (United States)

    Kim, Tae Yun; Song, Kye-Yong; Sohn, Woon-Mok; Kang, Shin-Yong

    2001-01-01

    In the course of Clonorchis sinensis infection, antigens presented to the hosts may be in a close relation to growth of the fluke. The antigenic proteins stimulating IgG antibody production were chronologically identified by immunoblot and localized by immunohistochemical staining. In the early stage of infection until 12 weeks post-infection (PI), antigens were proteins with molecular mass larger than 34 kDa which were derived from the tegument, testes and intrauterine eggs. After 20 weeks PI, antigens recognized were 29, 27 and 26 kDa proteins from the intestine, excretory bladder and reproductive organs. It is suggested that the tegumental proteins are the most potent antigens and the excretory-secretory proteins with middle molecular mass of 26-45 kDa contribute to the high level production of antibodies after 20 weeks of the C. sinensis infection. PMID:11775331

  7. Surface topography of two trematodes parasites infecting grey heron Ardea cinerea Jouyi (Aves, Ciconiiformes) in Qena, Egypt.

    Science.gov (United States)

    Ammar, Khalaf Nour Abd El-Wahed

    2015-04-01

    Apharyngostrigea ardeolina and Echinoparyphium recurvatum are two important digenean parasites that were recovered from small intestine of grey heron with an infection rate (16.2%) and (8.8%) respectively. The surface topography of two species was redescribed by both light and scanning electron microscopy. Using SEM studies showed that the body surface of two trematodes were covered by contact receptors, several types of sensory tegumental papillae which may have useful function in orientation and feeding through increasing the surface area of absorption, could also play a role in sensation or in selection of the materials for ingestion by the fluke. The head collar of E. recurvatum is reniform in shape, bearing uninterrupted double row of 41 collar finger-like spines, a total including 4 end group ones on both ventral corners., tegumental spines were tongue-shaped without a terminal tip.

  8. Isolation and risk assessment of Geotrichum spp. in the white shrimp (Litopenaeus vannamei Boone, 1931 from culture ponds

    Directory of Open Access Journals (Sweden)

    José Luis Ochoa

    2015-09-01

    Full Text Available The present study was done in order to identify the fungus invading some of the supralittoral ponds used for shrimp aquaculture in the CIBNOR facilities in La Paz, Baja California Sur (BCS, México during the summer season. From the walls and bottoms of the ponds, two strains of Geotrichum spp. were isolated and morphologically identified. Fungal adhesion towards hemocytes and primary cultures of various white shrimp (Litopeneaus vannamei tissues (gill, tegument, and gut was analyzed to determine infectivity. Extracellular protease, lipase, and amylase activity were evaluated as virulence factors. Survival of shrimp post-larvae (PL8 exposed to fungal culture supernatant or to their filaments was also investigated. The results showed that shrimp tegument cells and hemocytes were very susceptible to Geotrichum spp. invasion, and that this fungus provokes great mortality of post-larvae. Hence, Geotrichum spp. could be considered an opportunistic pathogen that might represent a serious health risk to shrimp in culture.

  9. Identification of pathogens for differential diagnosis of fever with jaundice in the Central African Republic: a retrospective assessment, 2008–2010

    OpenAIRE

    Gadia, Christelle Luce Bobossi; Manirakiza, Alexandre; Tekpa, Gaspard; Konamna, Xavier; Vickos, Ulrich; Nakoune, Emmanuel

    2017-01-01

    Background Febrile jaundice results clinically in generalized yellow coloration of the teguments and mucous membranes due to excess plasma bilirubin, accompanied by fever. Two types are found: conjugated and unconjugated bilirubin jaundice. Jaundice is a sign in several diseases due to viruses (viral hepatitis and arbovirus), parasites (malaria) and bacteria (leptospirosis). In the Central African Republic (CAR), only yellow fever is included on the list of diseases for surveillance. The aim ...

  10. WIDESPREAD CUTANEOUS LESIONS DUE TO Sporothrix schenckii IN A PATIENT UNDER A LONG TERM STEROIDS THERAPY

    Directory of Open Access Journals (Sweden)

    SEVERO Luiz Carlos

    1999-01-01

    Full Text Available A case of sporotrichosis in a woman presenting 63 cutaneous lesions distributed all over the tegument is related. The patient had both humoral (Immunoglobulins and cellular (Lymphocytes subpopulations immunity within normal limits, but was under treatment with steroid during a long time (Prednisone 10 mg daily for 2 years, due to a sciatic pain. In addition a review of the Brazilian literature on this type of lesions was carried out and commented.

  11. AcEST: BP915632 [AcEST

    Lifescience Database Archive (English)

    Full Text Available ... 31 2.6 sp|P03186|TEGU_EBV Large tegument protein OS=Epstein-Barr virus ... 30 3.3 sp|Q9E7N7|VP4A_LNYV Phosphoprotein OS=Lettuce...E7N7|VP4A_LNYV Phosphoprotein OS=Lettuce necrotic yellows virus GN=P PE=2 SV=1 Le

  12. The surface topography of Callorhynchocotyle callorhynchi (Manter, 1955) (Monogenea: Hexabothriidae), a parasite of the holocephalan fish Callorhinchus capensis.

    Science.gov (United States)

    Poddubnaya, Larisa G; Reed, Cecile; Gibson, David I

    2015-09-01

    A scanning electron microscopical study, incorporating some transmission electron microscopical observations, was undertaken on the surface topography of the gill parasite Callorhynchocotyle callorhynchi (Manter, 1955) (Monogenea: Hexabothriidae) from the Cape elephant fish Callorhinchus capensis (Holocephali) off the western coast of South Africa. The study revealed the presence of several new characteristics for this species. These include the presence of regularly distributed, knob-shaped projections on the surface of the haptor, haptoral appendix and sucker peduncles measuring 0.2 μm and in concentrations of approximately 100 per 10 μm(2) and the existence of a ridge which bisects each sucker lumen, forming two different loculi. We also report, for the first time for any monogenean, the presence of 'true spines'; these occur on the luminal surface of the haptoral suckers and have all of the characteristics of the tegumental spines of digeneans, i.e. they are situated within the distal syncytial tegumental cytoplasm, rest on the basal plasma membrane, have a uniform structure and are covered apically by the tegumental surface plasma membrane. These spines are simple, straight and single-pointed. Under the scanning electron microscopy (SEM), within an area of 20 μm in diameter, 23 such spines were counted, but their concentration and arrangement varies in different regions of the sucker. At their base, they measure about 1.5 μm in width and reach approximately 2 μm in height above the general level of the tegument, but transmission electron microscopy (TEM) measurements of an entire spine indicate that they may reach 3 μm in total length. The presence of spines, possessing similar morphological characteristics in both basal polyopisthocotylean monogeneans and digeneans, represents another characteristic which may prove useful in understanding the evolutionary relationships within the Neodermata.

  13. Glucose Uptake in the Human Pathogen Schistosoma mansoni Is Regulated Through Akt/Protein Kinase B Signaling.

    Science.gov (United States)

    McKenzie, Maxine; Kirk, Ruth S; Walker, Anthony J

    2018-06-05

    In Schistosoma mansoni, the facilitated glucose transporter SGTP4, which is expressed uniquely in the apical surface tegumental membranes of the parasite, imports glucose from host blood to support its growth, development, and reproduction. However, the molecular mechanisms that underpin glucose uptake in this blood fluke are not understood. In this study we employed techniques including Western blotting, immunolocalization, confocal laser scanning microscopy, pharmacological assays, and RNA interference to functionally characterize and map activated Akt in S mansoni. We find that Akt, which could be activated by host insulin and l-arginine, was active in the tegument layer of both schistosomules and adult worms. Blockade of Akt attenuated the expression and evolution of SGTP4 at the surface of the host-invading larval parasite life-stage, and suppressed SGTP4 expression at the tegument in adults; concomitant glucose uptake by the parasite was also attenuated in both scenarios. These findings shed light on crucial mechanistic signaling processes that underpin the energetics of glucose uptake in schistosomes, which may open up novel avenues for antischistosome drug development.

  14. AVALIAÇÃO DA ADAPTABILIDADE DE CAPRINOS AO SEMIÁRIDO ATRAVÉS DE PARÂMETROS FISIOLÓGICOS E ESTRUTURAS DO TEGUMENTO

    Directory of Open Access Journals (Sweden)

    ELISÂNGELA MARIA NUNES DA SILVA

    2010-01-01

    Full Text Available The experiment was carried with the objective of evaluating the adaptability of the breeds goats to Semiárid through physiologic parameters: rectal temperature, respiratory frequency and surface temperature and the structures of the tegument: perspiration glands, fat of glands and hair follicles. 40 female were used goat, of the breeds Anglo-Nubiana, Savana, Boer and Moxotó, being 10 animals of each breed, with medium age of five months. For the physiologic answers, he animals were allocated into a completely randomized design entirely in portions subdivided in the time, with the main treatments constituted by the breeds and the secondary ones by the shifts. For the study steorologic of the tegument was used in a completely randomized design entirely with four treatments and three repetitions. There was in significant teraction (P0.05 of the Boer breed. The breed Anglo-Nubiana it presented larger number of glands perspiration (P<0.05 and the breed Savanna presented the largest average of fat of glands (P<0.05. The studied breeds demonstrated to be physiological well adapted and with characteristics of the favorable tegument to the climatic conditions of semiarid.

  15. Antibody response in cattle, sheep and rats to infection with. gamma. -irradiated metacercariae of Fasciola hepatica

    Energy Technology Data Exchange (ETDEWEB)

    Hughes, D.L.; Doy, T.G. (Agricultural Research Council, Compton (UK). Inst. for Research on Animal Diseases); Hanna, R.E.B. (Queen' s Univ., Belfast, Northern Ireland (UK))

    1982-05-01

    Cattle, sheep and rats were infected orally with ..gamma..-irradiated metacercariae of Fasciola hepatica, or with normal metacercariae. The antibody response was monitored in each host to metacercarial tegument (T0), juvenile tegument (T1), adult tegument (T2) and gut antigens. The response was examined at weekly intervals for cattle and sheep throughout 15 weeks of infection and four weeks after infection in rats, using an indirect fluorescent antibody labelling technique. It was found that the irradiated metacercariae engendered a normal humoral response to T0, T1 and gut antigens in all three hosts although the antibody levels were somewhat reduced due to early death or stunting of the flukes. T0 and T1 appeared to be antigenically similar. Antibodies against T2 appeared late in the animals infected with ..gamma..-irradiated metacercariae and the titres attained were considerably lower than in the controls. The T2 antigen stimulus in the animals given ..gamma..-irradiated metacercariae was probably provided by flukes which 'broke through' the developmental barrier imposed by irradiation and which were found alive at autopsy.

  16. Antibody response in cattle, sheep and rats to infection with γ-irradiated metacercariae of Fasciola hepatica

    International Nuclear Information System (INIS)

    Hughes, D.L.; Doy, T.G.

    1982-01-01

    Cattle, sheep and rats were infected orally with γ-irradiated metacercariae of Fasciola hepatica, or with normal metacercariae. The antibody response was monitored in each host to metacercarial tegument (T0), juvenile tegument (T1), adult tegument (T2) and gut antigens. The response was examined at weekly intervals for cattle and sheep throughout 15 weeks of infection and four weeks after infection in rats, using an indirect fluorescent antibody labelling technique. It was found that the irradiated metacercariae engendered a normal humoral response to T0, T1 and gut antigens in all three hosts although the antibody levels were somewhat reduced due to early death or stunting of the flukes. T0 and T1 appeared to be antigenically similar. Antibodies against T2 appeared late in the animals infected with γ-irradiated metacercariae and the titres attained were considerably lower than in the controls. The T2 antigen stimulus in the animals given γ-irradiated metacercariae was probably provided by flukes which 'broke through' the developmental barrier imposed by irradiation and which were found alive at autopsy. (author)

  17. Estabelecimento e crescimento in vitro de plântulas de louro-pardo

    Directory of Open Access Journals (Sweden)

    Tiago Antonio Fick

    2007-01-01

    Full Text Available The objectives of this work were to develop an in vitro establishment protocol and to quantify growth of louro-pardo (Cordia trichotoma (Vell. Arráb. Ex Steud. seedlings to produce asseptic explants. Seed imbibition and disinfection were studied for in vitro seedling establishment. Seeds were imbibed in distilled and autoclaved water for 0, 24, 48, 72, 96 or 120 h. Seeds without tegument were submerged in a 70% alcohol solution for 30 s and in a hypochloride solution of 2% or 5% for 0, 5, 10, 15 or 20 min. Percentages of disinfection and germination and mean germination time were evaluated. Seedling growth of louro-pardo was quantified in 1/2MS and WPM culture mediums. Number of emerged leaves and roots and length of shoot and primary root were evaluated at 7, 14, 21 and 28 days after inoculation. Imbibition of louro-pardo seeds until 24 hours made easy tegument excision without affecting disinfection and germination. Seed-tegument excision and immersion in a 70% alcohol solution for 30 s were enough for an acceptable production of in vitro asseptic seedlings. Louro-pardo seedlings grow satisfactorily in a WPM culture medium without growth regulators.

  18. Quantitative Evaluation of Protein Heterogeneity within Herpes Simplex Virus 1 Particles.

    Science.gov (United States)

    El Bilali, Nabil; Duron, Johanne; Gingras, Diane; Lippé, Roger

    2017-05-15

    Several virulence genes have been identified thus far in the herpes simplex virus 1 genome. It is also generally accepted that protein heterogeneity among virions further impacts viral fitness. However, linking this variability directly with infectivity has been challenging at the individual viral particle level. To address this issue, we resorted to flow cytometry (flow virometry), a powerful approach we recently employed to analyze individual viral particles, to identify which tegument proteins vary and directly address if such variability is biologically relevant. We found that the stoichiometry of the U L 37, ICP0, and VP11/12 tegument proteins in virions is more stable than the VP16 and VP22 tegument proteins, which varied significantly among viral particles. Most interestingly, viruses sorted for their high VP16 or VP22 content yielded modest but reproducible increases in infectivity compared to their corresponding counterparts containing low VP16 or VP22 content. These findings were corroborated for VP16 in short interfering RNA experiments but proved intriguingly more complex for VP22. An analysis by quantitative Western blotting revealed substantial alterations of virion composition upon manipulation of individual tegument proteins and suggests that VP22 protein levels acted indirectly on viral fitness. These findings reaffirm the interdependence of the virion components and corroborate that viral fitness is influenced not only by the genome of viruses but also by the stoichiometry of proteins within each virion. IMPORTANCE The ability of viruses to spread in animals has been mapped to several viral genes, but other factors are clearly involved, including virion heterogeneity. To directly probe whether the latter influences viral fitness, we analyzed the protein content of individual herpes simplex virus 1 particles using an innovative flow cytometry approach. The data confirm that some viral proteins are incorporated in more controlled amounts, while

  19. Ada Apa Dengan SAP (AADS Akrual?

    Directory of Open Access Journals (Sweden)

    Eka Findi Tresnawati

    2013-08-01

    Full Text Available This article provides anoverview regarding SAP accrual-based and its problems. It compares information content of PP 24/2005’s cash toward accrual and PP 71/2010’s accrual-based toshow the urgency of accrual-based financial reporting requirements. The analysis borrows Abeysekara’s accounting imperialism. Discussion also involves empirical evidence and evaluation of accrual-based implementation probability in Sumenep Regency. Findings show that accrual-based information is not an urgent need and tends to be difficult to be implemented. Reflecting the evidence in Sumenep, local governments are faced with the need of human resources, the question of the use-fulness of accrual information, and technical difficulties on implementation.

  20. The herpes simplex virus 1 UL51 protein interacts with the UL7 protein and plays a role in its recruitment into the virion.

    Science.gov (United States)

    Roller, Richard J; Fetters, Rachel

    2015-03-01

    The alphaherpesvirus UL51 protein is a tegument component that interacts with the viral glycoprotein E and functions at multiple steps in virus assembly and spread in epithelial cells. We show here that pUL51 forms a complex in infected cells with another conserved tegument protein, pUL7. This complex can form in the absence of other viral proteins and is largely responsible for recruitment of pUL7 to cytoplasmic membranes and into the virion tegument. Incomplete colocalization of pUL51 and pUL7 in infected cells, however, suggests that a significant fraction of the population of each protein is not complexed with the other and that they may accomplish independent functions. The ability of herpesviruses to spread from cell to cell in the face of an immune response is critical for disease and shedding following reactivation from latency. Cell-to-cell spread is a conserved ability of herpesviruses, and the identification of conserved viral genes that mediate this process will aid in the design of attenuated vaccines and of novel therapeutics. The conserved UL51 gene of herpes simplex virus 1 plays important roles in cell-to-cell spread and in virus assembly in the cytoplasm, both of which likely depend on specific interactions with other viral and cellular proteins. Here we identify one of those interactions with the product of another conserved herpesvirus gene, UL7, and show that formation of this complex mediates recruitment of UL7 to membranes and to the virion. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  1. Morphological Characteristics of Schistosoma mansoni PZQ-Resistant and -Susceptible Strains Are Different in Presence of Praziquantel

    Science.gov (United States)

    Pinto-Almeida, António; Mendes, Tiago; de Oliveira, Rosimeire Nunes; Corrêa, Sheila de Andrade Penteado; Allegretti, Silmara Marques; Belo, Silvana; Tomás, Ana; Anibal, Fernanda de Freitas; Carrilho, Emanuel; Afonso, Ana

    2016-01-01

    Schistosomiasis is one of the most common human parasitic diseases whose socioeconomic impact is only surpassed by malaria. Praziquantel (PZQ) is the only drug commercially available for the treatment of all schistosome species causing disease in humans. However, there has been stronger evidences of PZQ-resistance on Schistosoma mansoni and thus it is very important to study the phenotypic characteristics associated with it. The aim of this study was to evaluate morphological alterations in S. mansoni PZQ-resistant adult worms and eggs, by comparing a PZQ- resistant strain obtained under PZQ drug pressure with a PZQ-susceptible strain. For this, scanning electronic microscopy was used to assess tegumental responsiveness of both strains under PZQ exposure, and optical microscopy allowed the monitoring of worms and eggs in the presence of the drug. Those assays showed that PZQ-susceptible worms exposed to the drug had more severe tegumental damages than the resistant one, which had only minor alterations. Moreover, contrary to what occurred in the susceptible strain, resistant worms were viable after PZQ exposure and gradually regaining full motility after removal of the drug. Eggs from resistant strain parasites are considerably smaller than those from susceptible strain. Our results suggest that there might be a difference in the tegument composition of the resistant strain and that worms are less responsive to PZQ. Changes observed in egg morphology might imply alterations in the biology of schistosomes associated to PZQ-resistance, which could impact on transmission and pathology of the disease. Moreover, we propose a hypothetical scenario where there is a different egg tropism of the S. mansoni resistant strain. This study is the first comparing two strains that only differ in their resistance characteristics, which makes it a relevant step in the search for resistance determinants. PMID:27199925

  2. A 3D model of the membrane protein complex formed by the white spot syndrome virus structural proteins.

    Directory of Open Access Journals (Sweden)

    Yun-Shiang Chang

    Full Text Available BACKGROUND: Outbreaks of white spot disease have had a large negative economic impact on cultured shrimp worldwide. However, the pathogenesis of the causative virus, WSSV (whit spot syndrome virus, is not yet well understood. WSSV is a large enveloped virus. The WSSV virion has three structural layers surrounding its core DNA: an outer envelope, a tegument and a nucleocapsid. In this study, we investigated the protein-protein interactions of the major WSSV structural proteins, including several envelope and tegument proteins that are known to be involved in the infection process. PRINCIPAL FINDINGS: In the present report, we used coimmunoprecipitation and yeast two-hybrid assays to elucidate and/or confirm all the interactions that occur among the WSSV structural (envelope and tegument proteins VP51A, VP19, VP24, VP26 and VP28. We found that VP51A interacted directly not only with VP26 but also with VP19 and VP24. VP51A, VP19 and VP24 were also shown to have an affinity for self-interaction. Chemical cross-linking assays showed that these three self-interacting proteins could occur as dimers. CONCLUSIONS: From our present results in conjunction with other previously established interactions we construct a 3D model in which VP24 acts as a core protein that directly associates with VP26, VP28, VP38A, VP51A and WSV010 to form a membrane-associated protein complex. VP19 and VP37 are attached to this complex via association with VP51A and VP28, respectively. Through the VP26-VP51C interaction this envelope complex is anchored to the nucleocapsid, which is made of layers of rings formed by VP664. A 3D model of the nucleocapsid and the surrounding outer membrane is presented.

  3. Histological damage and inflammatory response elicited by Monobothrium wageneri (Cestoda in the intestine of Tinca tinca (Cyprinidae

    Directory of Open Access Journals (Sweden)

    Sayyaf Dezfuli Bahram

    2011-12-01

    Full Text Available Abstract Background Among the European cyprinids, tench, Tinca tinca (L., and the pathological effects their cestodes may effect, have received very little or no attention. Most literature relating to Monobothrium wageneri Nybelin, 1922, a common intestinal cestode of tench, for example, has focused on aspects of its morphology rather than on aspects of the host-parasite interaction. Results Immunopathological and ultrastructural studies were conducted on the intestines of 28 tench, collected from Lake Piediluco, of which 16 specimens harboured tight clusters of numerous M. wageneri attached to the intestinal wall. The infection was associated with the degeneration of the mucosal layer and the formation of raised inflammatory swelling surrounding the worms. At the site of infection, the number of granulocytes in the intestine of T. tinca was significantly higher than the number determined 1 cm away from the site of infection or the number found in uninfected fish. Using transmission electron microscopy, mast cells and neutrophils were frequently observed in close proximity to, and inside, the intestinal capillaries; often these cells were in contact with the cestode tegument. At the host-parasite interface, no secretion from the parasite's tegument was observed. Intense degranulation of the mast cells was seen within the submucosa and lamina muscularis, most noticeably at sites close to the tegument of the scolex. In some instances, rodlet cells were encountered in the submucosa. In histological sections, hyperplasia of the mucous cells, notably those giving an alcian blue positive reaction, were evident in the intestinal tissues close to the swelling surrounding the worms. Enhanced mucus secretion was recorded in the intestines of infected tench. Conclusions The pathological changes and the inflammatory cellular response induced by the caryophyllidean monozoic tapeworm M. wageneri within the intestinal tract of an Italian population of wild

  4. (Vellozo Rohwer (Canela-sassafrás

    Directory of Open Access Journals (Sweden)

    Ricardo Cetnarski Filho

    2005-01-01

    Full Text Available The experiment was developed at the Seed Laboratory of Forestry Sciences Department of Federal University of Paraná – UFPR, in order to determinate the effect of the temperature: 20, 25 and 30o C, and the remotion of the outer coat (endocarp and tegument on seeds germination the Ocotea odorifera (Vel. Rohwer. The diaspores were collected at Fazenda Rio Grande, Paraná State, in a remaining Ombrophylous Mixed Forest. It was adopted the entirely randomized statistical model, The diaspores without outer coat presented a higher germination percentage and germination speed.

  5. First evidence of the presence of microtriches in the Gyrocotylidea

    Czech Academy of Sciences Publication Activity Database

    Poddubnaya, L. G.; Bruňanská, Magdaléna; Kuchta, Roman; Scholz, Tomáš

    2006-01-01

    Roč. 92, č. 4 (2006), s. 703-707 ISSN 0022-3395 R&D Projects: GA ČR GA206/03/1317; GA ČR GA524/04/0342; GA ČR GD524/03/H133; GA MŠk LC522 Grant - others:Russian Foundation of Fundamental Research(RU) 05-04-48250 Institutional research plan: CEZ:AV0Z60220518 Keywords : ultrastructure * tegument * phylogeny Subject RIV: EA - Cell Biology Impact factor: 1.300, year: 2006

  6. [The morphology of the causative agent of human diphyllobothriasis in the Far East].

    Science.gov (United States)

    Dovgalev, A S; Valovaia, M A; Piskunova, Iu A; Romanenko, N A; Khodakova, V I; Artamoshin, A S

    1991-01-01

    The morphology of types F, A, C plerocercoids, and mature Diphyllobothriidae of the Far Eastern populations (USSR) was studied by using scanning electron microscopy. New data were obtained on structure the skin muscular sac of plerocercoids type F (presence of tegumental microvilli, multi poly-layer of longitudinal subtegumental musculature, and variation of quantity of muscular filaments along strobila. The morphological identity of mature Diphyllobothriidae from the volunteer (self-infection of plerocercoid "type F"), infested men and seagulls from focuses of diphyllobothriasis in the Pacific Ocean regions in the Far East of the USSR was proved.

  7. Comparison of two procedures for labelling the surface of the hydatid disease organism, Echinococcus granulosus, with /sup 125/I

    Energy Technology Data Exchange (ETDEWEB)

    McManus, D.P.; McLaren, D.J.; Clark, N.W.T.; Parkhouse, R.M.E.

    1987-03-01

    Living, intact protoscoleces of the British horse and sheep strains of Echinococcus granulosus were subjected to surface radioiodination procedures using /sup 125/I and Iodogen and /sup 125/I-Bolton Hunter reagent. Subsequent combined electron microscopy and autoradiography revealed specific surface membrane labelling with the Iodogen procedure, but significant tegumental labelling with the Bolton-Hunter reagent. The two parasite strains yielded different profiles of electrophoretically separated labelled proteins; the Iodogen method, not surprisingly, resulted in a less complex pattern of labelled polypeptides than the Bolton and Hunter reagent.

  8. A comparison of two procedures for labelling the surface of the hydatid disease organism, Echinococcus granulosus, with 125I

    International Nuclear Information System (INIS)

    McManus, D.P.; McLaren, D.J.; Clark, N.W.T.; Parkhouse, R.M.E.

    1987-01-01

    Living, intact protoscoleces of the British horse and sheep strains of Echinococcus granulosus were subjected to surface radioiodination procedures using 125 I and Iodogen and 125 I-Bolton Hunter reagent. Subsequent combined electron microscopy and autoradiography revealed specific surface membrane labelling with the Iodogen procedure, but significant tegumental labelling with the Bolton-Hunter reagent. The two parasite strains yielded different profiles of electrophoretically separated labelled proteins; the Iodogen method, not surprisingly, resulted in a less complex pattern of labelled polypeptides than the Bolton and Hunter reagent. (author)

  9. The in vitro anthelmintic effects of plumbagin on newly excysted and 4-weeks-old juvenile parasites of Fasciola gigantica.

    Science.gov (United States)

    Lorsuwannarat, Natcha; Piedrafita, David; Chantree, Pathanin; Sansri, Veerawat; Songkoomkrong, Sineenart; Bantuchai, Sirasate; Sangpairot, Kant; Kueakhai, Pornanan; Changklungmoa, Narin; Chaichanasak, Pannigan; Chansela, Piyachat; Sobhon, Prasert

    2014-01-01

    The effect of plumbagin (PB, 5-hydroxy-2-methyl-1,4-naphthoquinone) against newly excysted juveniles (NEJs) and 4-weeks-old immature parasites of Fasciola gigantica were compared with triclabendazole (TCZ). The anthelmintic efficacy of 1, 10 and 100μg/ml of PB or TCZ following incubation in vitro for 1-24h was compared using a combination of relative motility (RM), survival index (SI) and larval migration inhibition (LMI) assays for parasite viability. The RM and SI values of the PB-treated group decreased at a more rapid rate than the TCZ-treated group. For NEJs, the decreased RM values were first observed at 1h incubation with 1μg/ml PB, and 90% of flukes were killed at 24h. In contrast, in TCZ-treated groups a 10-fold higher concentration of TCZ (10μg/ml) resulted in only 9% dead parasites after 24h incubation. In 4-weeks-old juvenile parasites, PB reduced the RM value at 10μg/ml with 100% of flukes dead after 3h, while TCZ decreased RM values at the concentration of 100μg/ml but with only 5% of flukes killed at 24h. NEJs treated with PB exhibited 88%, 99% and 100% of LMIs at the concentrations of 1, 10 and 100μg/ml, respectively. NEJs incubated with TCZ have an LMI of only 32% at the highest concentration of 100μg/ml. Similarly PB had a significantly greater killing of immature 4weeks juvenile stages than TCZ at all concentrations; however, 4-weeks-old juvenile parasites were more resistant to killing by PB or TCZ at all concentrations when compared to NEJs. Further studies were carried out to investigate the alterations of the parasite tegument by scanning electron microscope (SEM). PB caused similar tegumental alterations in 4-weeks-old juveniles as those observed in TCZ treatment but with greater damage at comparative time points, comprising of swelling, blebbing and rupture of the tegument, loss of spines, and eventual erosion, lesion and desquamation of the total tegument. These data indicate that PB had a greater fasciolicidal effect against

  10. Identification of immediate early gene products of bovine herpes virus 1 (BHV-1) as dominant antigens recognized by CD8 T cells in immune cattle

    DEFF Research Database (Denmark)

    Hart, Jane; MacHugh, Niall D.; Sheldrake, Tara

    2017-01-01

    candidate viral gene products with CD8 T-cell lines from 3 BHV-1-immune cattle of defined MHC genotypes identified 4 antigens, including 3 immediate early (IE) gene products (ICP4, ICP22 and Circ) and a tegument protein (UL49). Identification of the MHC restriction specificities revealed that the antigens...... cases refined, the identity of the epitopes. Analyses of the epitope specificity of the CD8 T-cell lines showed that a large component of the response is directed against these IE epitopes. The results indicate that these IE gene products are dominant targets of the CD8 T-cell response in BHV...

  11. Crystal Structure of the N-Terminal Half of the Traffic Controller UL37 from Herpes Simplex Virus 1

    Energy Technology Data Exchange (ETDEWEB)

    Koenigsberg, Andrea L.; Heldwein, Ekaterina E.; Sandri-Goldin, Rozanne M.

    2017-08-02

    Inner tegument protein UL37 is conserved among all three subfamilies of herpesviruses. Studies of UL37 homologs from two alphaherpesviruses, herpes simplex virus 1 (HSV-1) and pseudorabies virus (PRV), have suggested that UL37 plays an essential albeit poorly defined role in intracellular capsid trafficking. At the same time, HSV and PRV homologs cannot be swapped, which suggests that in addition to a conserved function, UL37 homologs also have divergent virus-specific functions. Accurate dissection of UL37 functions requires detailed maps in the form of atomic-resolution structures. Previously, we reported the crystal structure of the N-terminal half of UL37 (UL37N) from PRV. Here, we report the crystal structure of HSV-1 UL37N. Comparison of the two structures reveals that UL37 homologs differ in their overall shapes, distributions of surface charges, and locations of projecting loops. In contrast, the previously identified R2 surface region is structurally conserved. We propose that within the N-terminal half of UL37, functional conservation is centered within the R2 surface region, whereas divergent structural elements pinpoint regions mediating virus-specific functions and may engage different binding partners. Together, the two structures can now serve as templates for a structure-guided exploration of both conserved and virus-specific functions of UL37.

    IMPORTANCEThe ability to move efficiently within host cell cytoplasm is essential for replication in all viruses. It is especially important in the neuroinvasive alphaherpesviruses, such as human herpes simplex virus 1 (HSV-1), HSV-2, and veterinarian pseudorabies virus (PRV), that infect the peripheral nervous system and have to travel long distances along axons. Capsid movement in these viruses is controlled by capsid-associated tegument proteins, yet their specific roles have not yet been defined. Systematic exploration of the roles of tegument proteins in capsid trafficking requires

  12. Protein kinase C and extracellular signal-regulated kinase regulate movement, attachment, pairing and egg release in Schistosoma mansoni.

    Directory of Open Access Journals (Sweden)

    Margarida Ressurreição

    2014-06-01

    Full Text Available Protein kinases C (PKCs and extracellular signal-regulated kinases (ERKs are evolutionary conserved cell signalling enzymes that coordinate cell function. Here we have employed biochemical approaches using 'smart' antibodies and functional screening to unravel the importance of these enzymes to Schistosoma mansoni physiology. Various PKC and ERK isotypes were detected, and were differentially phosphorylated (activated throughout the various S. mansoni life stages, suggesting isotype-specific roles and differences in signalling complexity during parasite development. Functional kinase mapping in adult worms revealed that activated PKC and ERK were particularly associated with the adult male tegument, musculature and oesophagus and occasionally with the oesophageal gland; other structures possessing detectable activated PKC and/or ERK included the Mehlis' gland, ootype, lumen of the vitellaria, seminal receptacle and excretory ducts. Pharmacological modulation of PKC and ERK activity in adult worms using GF109203X, U0126, or PMA, resulted in significant physiological disturbance commensurate with these proteins occupying a central position in signalling pathways associated with schistosome muscular activity, neuromuscular coordination, reproductive function, attachment and pairing. Increased activation of ERK and PKC was also detected in worms following praziquantel treatment, with increased signalling associated with the tegument and excretory system and activated ERK localizing to previously unseen structures, including the cephalic ganglia. These findings support roles for PKC and ERK in S. mansoni homeostasis, and identify these kinase groups as potential targets for chemotherapeutic treatments against human schistosomiasis, a neglected tropical disease of enormous public health significance.

  13. SCHISTOSOMICIDAL ACTIVITY OF THE CRUDE EXTRACT OF ARTOCARPUS LAKOOCHA.

    Science.gov (United States)

    Preyavichyapugdee, Narin; Sangfuang, Manaw; Chaiyapum, Saowapark; Sriburin, Suwatcharaporn; Pootaeng-on, Yupa; Chusongsang, Phiraphol; Jiraungkoorskul, Wannee; Preyavichyapugdee, Mananya; Sobhon, Prasert

    2016-01-01

    Puag-Haad is a traditional anthelmintic drug used to treat taeniasis in Thailand and Lao PDR. It is derived from the aqueous extract of the plant Artocarpus lakoocha. We investigated the in vitro anthelmintic properties of Puag-Haad against Schistosoma mansoni. Adult worms were incubated in M-199 medium containing 250, 500 and 750 μg/ml of Puag-Haad or praziquantel (PZQ) at a concentration of 175 μg/ml for 3, 6, 12 and 24 hours. The relative motility (RM value), survival index (SI) and tegument alterations seen under scanning electron microscope were assessed at each incubation time. The results showed the crude extract of A. lakoocha at a concentration of 250 μg/ml was more effective in causing damage than PZQ at a concentration of 175 μg/ml using RM and SI values. The major target organ affected by Puag-Haad was the tegument. The damage was greater at higher concentrations of the crude extract. It is likely tetrahydroxystilbene (THS), the main compound in Puag-Haad, caused the damage. THS could be a future candidate as a schistosomal drug. Further studies are needed to explore its mechanism, efficiency and safety in vivo.

  14. Proteomic analysis of the herpes simplex virus 1 virion protein 16 transactivator protein in infected cells.

    Science.gov (United States)

    Suk, Hyung; Knipe, David M

    2015-06-01

    The herpes simplex virus 1 virion protein 16 (VP16) tegument protein forms a transactivation complex with the cellular proteins host cell factor 1 (HCF-1) and octamer-binding transcription factor 1 (Oct-1) upon entry into the host cell. VP16 has also been shown to interact with a number of virion tegument proteins and viral glycoprotein H to promote viral assembly, but no comprehensive study of the VP16 proteome has been performed at early times postinfection. We therefore performed a proteomic analysis of VP16-interacting proteins at 3 h postinfection. We confirmed the interaction of VP16 with HCF-1 and a large number of cellular Mediator complex proteins, but most surprisingly, we found that the major viral protein associating with VP16 is the infected cell protein 4 (ICP4) immediate-early (IE) transactivator protein. These results raise the potential for a new function for VP16 in associating with the IE ICP4 and playing a role in transactivation of early and late gene expression, in addition to its well-documented function in transactivation of IE gene expression. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. New perspectives on host-parasite interplay by comparative transcriptomic and proteomic analyses of Schistosoma japonicum.

    Directory of Open Access Journals (Sweden)

    Feng Liu

    2006-04-01

    Full Text Available Schistosomiasis remains a serious public health problem with an estimated 200 million people infected in 76 countries. Here we isolated ~ 8,400 potential protein-encoding cDNA contigs from Schistosoma japonicum after sequencing circa 84,000 expressed sequence tags. In tandem, we undertook a high-throughput proteomics approach to characterize the protein expression profiles of a number of developmental stages (cercariae, hepatic schistosomula, female and male adults, eggs, and miracidia and tissues at the host-parasite interface (eggshell and tegument by interrogating the protein database deduced from the contigs. Comparative analysis of these transcriptomic and proteomic data, the latter including 3,260 proteins with putative identities, revealed differential expression of genes among the various developmental stages and sexes of S. japonicum and localization of putative secretory and membrane antigens, enzymes, and other gene products on the adult tegument and eggshell, many of which displayed genetic polymorphisms. Numerous S. japonicum genes exhibited high levels of identity with those of their mammalian hosts, whereas many others appeared to be conserved only across the genus Schistosoma or Phylum Platyhelminthes. These findings are expected to provide new insights into the pathophysiology of schistosomiasis and for the development of improved interventions for disease control and will facilitate a more fundamental understanding of schistosome biology, evolution, and the host-parasite interplay.

  16. Varicella-zoster virus induces the formation of dynamic nuclear capsid aggregates

    Energy Technology Data Exchange (ETDEWEB)

    Lebrun, Marielle [University of Liege (ULg), GIGA-Infection Immunity and Inflammation, Laboratory of Virology and Immunology, Liege (Belgium); Thelen, Nicolas; Thiry, Marc [University of Liege (ULg), GIGA-Neurosciences, Laboratory of Cellular and Tissular Biology, Liege (Belgium); Riva, Laura; Ote, Isabelle; Condé, Claude; Vandevenne, Patricia [University of Liege (ULg), GIGA-Infection Immunity and Inflammation, Laboratory of Virology and Immunology, Liege (Belgium); Di Valentin, Emmanuel [University of Liege (ULg), GIGA-Viral Vectors Platform, Liege (Belgium); Bontems, Sébastien [University of Liege (ULg), GIGA-Infection Immunity and Inflammation, Laboratory of Virology and Immunology, Liege (Belgium); Sadzot-Delvaux, Catherine, E-mail: csadzot@ulg.ac.be [University of Liege (ULg), GIGA-Infection Immunity and Inflammation, Laboratory of Virology and Immunology, Liege (Belgium)

    2014-04-15

    The first step of herpesviruses virion assembly occurs in the nucleus. However, the exact site where nucleocapsids are assembled, where the genome and the inner tegument are acquired, remains controversial. We created a recombinant VZV expressing ORF23 (homologous to HSV-1 VP26) fused to the eGFP and dually fluorescent viruses with a tegument protein additionally fused to a red tag (ORF9, ORF21 and ORF22 corresponding to HSV-1 UL49, UL37 and UL36). We identified nuclear dense structures containing the major capsid protein, the scaffold protein and maturing protease, as well as ORF21 and ORF22. Correlative microscopy demonstrated that the structures correspond to capsid aggregates and time-lapse video imaging showed that they appear prior to the accumulation of cytoplasmic capsids, presumably undergoing the secondary egress, and are highly dynamic. Our observations suggest that these structures might represent a nuclear area important for capsid assembly and/or maturation before the budding at the inner nuclear membrane. - Highlights: • We created a recombinant VZV expressing the small capsid protein fused to the eGFP. • We identified nuclear dense structures containing capsid and procapsid proteins. • Correlative microscopy showed that the structures correspond to capsid aggregates. • Procapsids and partial capsids are found within the aggregates of WT and eGFP-23 VZV. • FRAP and FLIP experiments demonstrated that they are dynamic structures.

  17. An ultrastructural examination of irradiated, immunizing schistosomula of Schistosoma mansoni during their extended stay in the lungs

    International Nuclear Information System (INIS)

    Mastin, A.; Wilson, R.A.; Bickle, Q.D.

    1985-01-01

    An ultrastructural study has been performed on radiation-attenuated parasites fixed in situ during their extended stay in the lungs. At day 7 post-infection all parasites were located within blood vessels. In contrast to normally migrating parasites, at days 13 and 21 post-infection the majority of radiation-attenuated parasites were lying free within alveoli. It appeared that radiation-attenuated parasites had undergone the normal developmental changes associated with the lung-stage of migration. No ultrastructural evidence was found to suggest why schistosomula failed to migrate further and instead burst into alveoli. No parasites observed on day 7 post-infection were associated with a host inflammatory reaction. At day 13, inflammation was noticeable but could have been a response as much to pulmonary tissue damage as to the presence of parasites. At day 21 post-infection some parasites were internally disrupted but were not associated with host inflammatory reactions and had intact tegumental surfaces. One parasite was observed at day 21 with an adherent neutrophil infiltrate and extensive tegumental damage. The relevance of persisting lung-stage parasites to the induction of resistance to challenge is discussed. (author)

  18. [Prokaryotic expression and histological localization of the Taenia solium CDC37 gene].

    Science.gov (United States)

    Huang, Jiang; Li, Bo; Dai, Jia-Lin; Zhang, Ai-Hua

    2013-02-01

    To express Taenia solium gene encoding cell division cycle 37 protein (TsCDC37) and investigate its antigenicity and localization in adults of Taenia solium. The complete coding sequence of TsCDC37 was amplified by PCR based on the recombinant plasmid clone from the cDNA library of adult Taenia solium. The PCR product was cloned into a prokaryotic expression vector pET-28a (+). The recombinant expression plasmid was identified by PCR, double endonuclease digestion and sequencing. The recombinant plasmid was transformed into E. coli BL21/DE3 and followed by expression of the protein induced by IPTG. The mice were immunized subcutaneously with purified recombinant TsCDC37 formulated in Freund's adjuvant. The antigenicity of the recombinant protein was examined by Western blotting. The localization of TsCDC37 in adult worms was demonstrated by immunofluorescent technique. The recombinant expression vector was constructed successfully. The recombinant protein was about M(r) 52 000, it was then purified and specifically recognized by immuno sera of SD rats and sera from patients infected with Taenia solium, Taenia saginata or Taenia asiatica. The immunofluorescence assay revealed that TsCDC37 located at the tegument of T. solium adult and the eggs. TsCDC37 gene has been expressed with immunoreactivity. The recombinant protein is mainly expressed in tegument and egg, and is a common antigen of the three human taenia cestodes.

  19. INFLUÊNCIA DA COR E MÉTODOS DE SUPERAÇÃO DE DORMÊNCIA EM SEMENTES DE ALBIZIA

    Directory of Open Access Journals (Sweden)

    Clarisse Pereira Benedito

    2009-01-01

    Full Text Available The present work had as objective to evaluate the influence of pre-germinatives treatments to overcome dormancy in seeds of albizia (Albizia lebbeck with different colorations. It took place an experiment at greenhouse UFERSA, in Mossoró-RN, in completely randomized design with the treatments distributed in outline factorial 8x2, with the factors treatments for pre-germination and classes of seeds (clear seeds and dark seeds, in four replications of 25 seeds. The germinative pré-germinative consisted either in the immersion of the seeds in sulfuric acid for ten, 15 and 20 minutes; immersion in sodium hypochlorite for ten, 15 and 20 minutes; cutting in the tegument in the opposite side of the embryonic axis and the control (the control was represented by intact seeds. Were analyzed the following parameters: first germination count, germination speed index, and percentage of germination. . By considering the results, it was observed that the dark seeds were better than clear seed. The immersion treatments in sulfuric acid for 10 and 20 minutes and the cutting in the tegument were the most efficient for the break of seeds dormancy.

  20. The Extracellular Vesicles of the Helminth Pathogen, Fasciola hepatica: Biogenesis Pathways and Cargo Molecules Involved in Parasite Pathogenesis*

    Science.gov (United States)

    Cwiklinski, Krystyna; de la Torre-Escudero, Eduardo; Trelis, Maria; Bernal, Dolores; Dufresne, Philippe J.; Brennan, Gerard P.; O'Neill, Sandra; Tort, Jose; Paterson, Steve; Marcilla, Antonio; Dalton, John P.; Robinson, Mark W.

    2015-01-01

    Extracellular vesicles (EVs) released by parasites have important roles in establishing and maintaining infection. Analysis of the soluble and vesicular secretions of adult Fasciola hepatica has established a definitive characterization of the total secretome of this zoonotic parasite. Fasciola secretes at least two subpopulations of EVs that differ according to size, cargo molecules and site of release from the parasite. The larger EVs are released from the specialized cells that line the parasite gastrodermus and contain the zymogen of the 37 kDa cathepsin L peptidase that performs a digestive function. The smaller exosome-like vesicle population originate from multivesicular bodies within the tegumental syncytium and carry many previously described immunomodulatory molecules that could be delivered into host cells. By integrating our proteomics data with recently available transcriptomic data sets we have detailed the pathways involved with EV biogenesis in F. hepatica and propose that the small exosome biogenesis occurs via ESCRT-dependent MVB formation in the tegumental syncytium before being shed from the apical plasma membrane. Furthermore, we found that the molecular “machinery” required for EV biogenesis is constitutively expressed across the intramammalian development stages of the parasite. By contrast, the cargo molecules packaged within the EVs are developmentally regulated, most likely to facilitate the parasites migration through host tissue and to counteract host immune attack. PMID:26486420

  1. Evaluation of Protection against Schistosoma mansoni Infection in Mice Vaccinated with Irradiated Cercariae

    International Nuclear Information System (INIS)

    Amin, M.M.

    2009-01-01

    The Present study was designed to evaluate the biological changes in mice vaccinated with irradiated Schistosoma mansoni cercariae (40 krad) in comparison with infected non-vaccinated group. The degree of resistance was assessed by parasitological, biochemical, immunological, histopathological as well as scanning electron microscopy studies. The results of the present study revealed a significant reduction in worm burden and ova count in both liver and intestine of the immunized group. In addition, a moderate amelioration was recorded in the liver functions; gamma glutamyltransferase (gamma-GT), alkaline phosphatase (ALP), alanine and aspartate transaminase (ALT and AST). Assessment of tissue nuclear factor-a (TNF-a) and Interleukin -10 (IL-IO) in sera of the experimental groups showed significant protection changes. Histopathological examination of vaccinated mice livers showed protection against parasite maturation and liver damage after challenged as compared to mice infected only without vaccination. Also, scanning electron microscopy of whole worm revealed severe degree of surface tegumental disruption and intensive stunted of the worms as well as loss of the dorsal tegumental spines, On the other hand, there were severe deformations in both the anterior and ventral suckers as a result of the immunization

  2. Herpes Simplex Virus 1 Us3 Deletion Mutant is Infective Despite Impaired Capsid Translocation to the Cytoplasm

    Directory of Open Access Journals (Sweden)

    Peter Wild

    2015-01-01

    Full Text Available Herpes simplex virus 1 (HSV-1 capsids are assembled in the nucleus bud at the inner nuclear membrane into the perinuclear space, acquiring envelope and tegument. In theory, these virions are de-enveloped by fusion of the envelope with the outer nuclear membrane and re-enveloped by Golgi membranes to become infective. Us3 enables the nucleus to cytoplasm capsid translocation. Nevertheless, Us3 is not essential for the production of infective progeny viruses. Determination of phenotype distribution by quantitative electron microscopy, and calculation per mean nuclear or cell volume revealed the following: (i The number of R7041(∆US3 capsids budding at the inner nuclear membrane was significantly higher than that of wild type HSV-1; (ii The mean number of R7041(∆US3 virions per mean cell volume was 2726, that of HSV-1 virions 1460 by 24 h post inoculation; (iii 98% of R7041(∆US3 virions were in the perinuclear space; (iv The number of R7041(∆US3 capsids in the cytoplasm, including those budding at Golgi membranes, was significantly reduced. Cell associated R7041(∆US3 yields were 2.37 × 108 and HSV-1 yields 1.57 × 108 PFU/mL by 24 h post inoculation. We thus conclude that R7041(∆US3 virions, which acquire envelope and tegument by budding at the inner nuclear membrane into the perinuclear space, are infective.

  3. Molecular confirmation of ovine herpesvirus 2-induced malignant catarrhal fever lesions in cattle from Rio Grande do Norte, Brazil

    Directory of Open Access Journals (Sweden)

    Selwyn A. Headley

    2012-12-01

    Full Text Available Molecular findings that confirmed the participation of ovine herpesvirus 2 (OVH-2 in the lesions that were consistent with those observed in malignant catarrhal fever of cattle are described. Three mixed-breed cattle from Rio Grande do Norte state demonstrated clinical manifestations that included mucopurulent nasal discharge, corneal opacity and motor incoordination. Routine necropsy examination demonstrated ulcerations and hemorrhage of the oral cavity, corneal opacity, and lymph node enlargement. Significant histopathological findings included widespread necrotizing vasculitis, non-suppurative meningoencephalitis, lymphocytic interstitial nephritis and hepatitis, and thrombosis. PCR assay performed on DNA extracted from kidney and mesenteric lymph node of one animal amplified a product of 423 base pairs corresponding to a target sequence within the ovine herpesvirus 2 (OVH-2 tegument protein gene. Direct sequencing of the PCR products, from extracted DNA of the kidney and mesenteric lymph node of one cow, amplified the partial nucleotide sequences (423 base pairs of OVH-2 tegument protein gene. Blast analysis confirmed that these sequences have 98-100% identity with similar OVH-2 sequences deposited in GenBank. Phylogenetic analyses, based on the deduced amino acid sequences, demonstrated that the strain of OVH-2 circulating in ruminants from the Brazilian states of Rio Grande do Norte and Minas Gerais are similar to that identified in other geographical locations. These findings confirmed the active participation of OVH-2 in the classical manifestations of sheep associated malignant catarrhal fever.

  4. Activity of praziquantel on in vitro transformed Schistosoma mansoni sporocysts

    Directory of Open Access Journals (Sweden)

    ACA Mattos

    2006-10-01

    Full Text Available Praziquantel (PZQ is effective against all the evolutive phases of Schistosoma mansoni. Infected Biomphalaria glabrata snails have their cercarial shedding interrupted when exposed to PZQ. Using primary in vitro transformed sporocysts, labeled with the probe Hoechst 33258 (indicator of membrane integrity, and lectin of Glycine max (specific for carbohydrate of N-acetylgalactosamine membrane, we evaluated the presence of lysosomes at this evolutive phase of S. mansoni, as well as the influence of PZQ on these acidic organelles and on the tegument of the sporocyst. Although the sporocyst remained alive, it was observed that there was a marked contraction of its musculature, and there occurred a change in the parasite's structure. Also, the acidic vesicles found in the sporocysts showed a larger delimited area after contact of the parasites with PZQ. Damages to the tegument was also observed, as show a well-marked labeling either with Hoechst 33258 or with lectin of Glycine max after contact of sporocysts with the drug. These results could partially explain the interruption/reduction mechanism of cercarial shedding in snails exposed to PZQ.

  5. Reproductive system abnormalities in Schistosoma mansoni adult worms isolated from Nectomys squamipes (Muridae: Sigmodontinae: brightfield and confocal laser scanning microscopy analysis

    Directory of Open Access Journals (Sweden)

    Neves Renata Heisler

    2003-01-01

    Full Text Available Schistosoma mansoni adult worms with genital anomalies isolated from Nectomys squamipes (Muridae: Sigmodontinae were studied by confocal laser scanning microscopy under the reflected mode. One male without testicular lobes (testicular agenesia/anorchism and two females, one with an atrophied ovary and another with 17 uterine eggs, were identified. The absence of testicular lobes occurred in a worm presenting otherwise normal male adult characteristics: tegument, tubercles and a gynaecophoric canal with spines. In both female specimens the digestive tube showed a vacuolated appearance, and the specimen with supernumerary uterine eggs exhibited a developing miracidium and an egg with a formed shell. The area of the ventral sucker was similar in both specimens however the tegument thickness, ovary and vitelline glands of the specimen with the atrophied ovary were smaller than those of the one with supernumerary eggs. These reported anomalies in the reproductive system call attention to the need to improve our understanding of genetic regulation and the possible role of environmental influences upon trematode development.

  6. Morphological aspects of Schistosoma mansoni adult worms isolated from nourished and undernourished mice: a comparative analysis by confocal laser scanning microscopy

    Directory of Open Access Journals (Sweden)

    Neves Renata Heisler

    2001-01-01

    Full Text Available Malnutrition hampers the course of schistosomiasis mansoni infection just as normal growth of adult worms. A comparative morphometric study on adult specimens (male and female recovered from undernourished (fed with a low protein diet - regional basic diet and nourished (rodent commercial laboratory food, NUVILAB white mice was performed. Tomographic images and morphometric analysis of the oral and ventral suckers, reproductive system and tegument were obtained by means of confocal laser scanning microscopy. Undernourished male specimens presented smaller morphometric values (length and width of the reproductive system (first, third and last testicular lobes and thickness of the tegument than controls. Besides that, it was demonstrated that the dorsal surface of the male worms bears large tubercles unevenly distributed, but kept grouped and flat. At the subtegumental region, vacuolated areas were detected. It was concluded that the inadequate nutritional status of the vertebrate host has a negative influence mainly in the reproductive system and topographical somatic development of male adult Schistosoma mansoni, inducing some alterations on the structure of the parasite.

  7. Anthelmintic Activity of Crude Extract and Essential Oil of Tanacetum vulgare (Asteraceae against Adult Worms of Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    Loyana Silva Godinho

    2014-01-01

    Full Text Available Schistosomiasis, a parasitic disease caused by trematode flatworms of the genus Schistosoma, affects more than 200 million people worldwide, and its control is dependent on a single drug, praziquantel. Tanacetum vulgare (Asteraceae is used in folk medicine as a vermifuge. This study aimed to investigate the in vitro schistosomicidal activity of the crude extract (TV and the essential oil (TV-EO from the aerial parts of T. vulgare. TV-EO was obtained by hydrodistillation and analyzed by GC/MS, which allowed the identification of β-thujone (84.13% as the major constituent. TV and TV-EO, at 200 μg/mL, decreased motor activity and caused 100% mortality of all adult worms. At 100 and 50 μg/mL, only TV caused death of all adult worms, while TV-EO was inactive. TV (200 μg/mL was also able to reduce viability and decrease production of developed eggs. Confocal laser scanning microscopy showed morphological alterations in the tegument of the S. mansoni surface after incubation with TV (50 and 100 μg/mL. Quantitative analysis on the schistosomes tegument showed that TV caused changes in the numbers of tubercles of S. mansoni male worms in a dose-dependent manner. The findings suggest that T. vulgare is a potential source of schistosomicidal compounds.

  8. Two-color fluorescence analysis of individual virions determines the distribution of the copy number of proteins in herpes simplex virus particles.

    Science.gov (United States)

    Clarke, Richard W; Monnier, Nilah; Li, Haitao; Zhou, Dejian; Browne, Helena; Klenerman, David

    2007-08-15

    We present a single virion method to determine absolute distributions of copy number in the protein composition of viruses and apply it to herpes simplex virus type 1. Using two-color coincidence fluorescence spectroscopy, we determine the virion-to-virion variability in copy numbers of fluorescently labeled tegument and envelope proteins relative to a capsid protein by analyzing fluorescence intensity ratios for ensembles of individual dual-labeled virions and fitting the resulting histogram of ratios. Using EYFP-tagged capsid protein VP26 as a reference for fluorescence intensity, we are able to calculate the mean and also, for the first time to our knowledge, the variation in numbers of gD, VP16, and VP22 tegument. The measurement of the number of glycoprotein D molecules was in good agreement with independent measurements of average numbers of these glycoproteins in bulk virus preparations, validating the method. The accuracy, straightforward data processing, and high throughput of this technique make it widely applicable to the analysis of the molecular composition of large complexes in general, and it is particularly suited to providing insights into virus structure, assembly, and infectivity.

  9. Operating Characteristics of Statistical Methods for Detecting Gene-by-Measured Environment Interaction in the Presence of Gene-Environment Correlation under Violations of Distributional Assumptions.

    Science.gov (United States)

    Van Hulle, Carol A; Rathouz, Paul J

    2015-02-01

    Accurately identifying interactions between genetic vulnerabilities and environmental factors is of critical importance for genetic research on health and behavior. In the previous work of Van Hulle et al. (Behavior Genetics, Vol. 43, 2013, pp. 71-84), we explored the operating characteristics for a set of biometric (e.g., twin) models of Rathouz et al. (Behavior Genetics, Vol. 38, 2008, pp. 301-315), for testing gene-by-measured environment interaction (GxM) in the presence of gene-by-measured environment correlation (rGM) where data followed the assumed distributional structure. Here we explore the effects that violating distributional assumptions have on the operating characteristics of these same models even when structural model assumptions are correct. We simulated N = 2,000 replicates of n = 1,000 twin pairs under a number of conditions. Non-normality was imposed on either the putative moderator or on the ultimate outcome by ordinalizing or censoring the data. We examined the empirical Type I error rates and compared Bayesian information criterion (BIC) values. In general, non-normality in the putative moderator had little impact on the Type I error rates or BIC comparisons. In contrast, non-normality in the outcome was often mistaken for or masked GxM, especially when the outcome data were censored.

  10. Design and Analysis of Financial Condition Local Government Java and Bali (2013-2014

    Directory of Open Access Journals (Sweden)

    Natrini Nur Dewi

    2017-01-01

    Full Text Available This research aims to identify financial condition of local government in Java and Bali year 2013-2014. It is due to government financial condition, according to several researchers, provides an image on the ability of a government in fulfilling their obligations whether in the form of debt or service fulfillment in timely manner. According to assessment upon financial condition, local government is able to identify how to fulfill public needs, how to utilize resources and how to proceed resources so that it can be more productive. As for the measurement method of financial condition itself, the standard method cannot be determined. Therefore, indicator used for measuring local government financial condition is Brown's (1993[2] indicator development adjusted to Indonesian government. In order to develop the indicator, this research employs qualitative method by comparing GASB No.34, SAP Government Regulation (“Peraturan Pemerintah” - PP 71 Year 2010, SAP PP 24 year 2005 and literature studies and expert validation. In order to obtain a balanced comparison, this research also employs clusters developed by Baidori (2015[1] for government in Java and Bali. Results of this research showed that among 7 analyzed clusters, there are variations of results, even though each cluster has similar socioeconomic condition to each other. This variation upon Indonesian local government financial condition is caused by regional autonomy.

  11. Influence of Schistosoma japonicum programmed cell death protein 10 on the growth and development of schistosomula.

    Science.gov (United States)

    Gao, Yan Ru; Huang, Wen Ling; Tang, Chun Lian; Liu, Rong; Zhao, Qin Ping; Ming, Zhen Ping; Dong, Hui Fen

    2018-01-18

    Schistosomiasis caused by Schistosoma japonicum is among the most serious endemic zoonoses in China. To study interactions between schistosomula, the pre-adult juvenile stage, and hosts, it is important to study the functions of key genes involved in schistosomula growth and development. Programmed cell death protein 10 (pcdp10) is an important apoptosis-related gene with various biological functions. This study described the molecular characterization of S. japonicum PCDP10 (SjPCDP10) and evaluated its functions in schistosomula. Real-time quantitative polymerase chain reaction (qPCR) and western blot were used to detect Sjpcdp10 mRNA and protein levels, respectively, at different developmental stages. Immunolocalization was performed to determine SjPCDP10 expression in the parasite. RNA interference (RNAi) experiments were used to assess gene functions associated with SjPCDP10 in schistosomula growth and development. Real-time qPCR revealed that Sjpcdp10 was expressed during all investigated developmental stages and upregulated during schistosomula growth and development. Histochemical localization showed that SjPCDP10 was mainly distributed in the teguments of schistosomula in all investigated stages and part of the parenchymal area of 14-, 18-, and 21-day-old schistosomula. Following Sjpcdp10 knockdown by RNAi, the lengths, widths, areas, and volumes of schistosomula were significantly lower than those in the control group. Scanning electron microscopy showed that the body surfaces of schistosomula subjected to RNAi were seriously damaged, with few tegumental spines and sensory papillae. Transmission electron microscopy indicated that the teguments of Sjpcdp10-knockdown schistosomula were incomplete, the number of layers was reduced, and the thickness decreased significantly as compared with those in the control group. Furthermore, terminal deoxynucleotidyl transferase dUTP nick-end labelling results showed that the rate of apoptosis in Sjpcdp10-knockdown

  12. Molecular phylogeny of equine herpesvirus 1 isolates from onager, zebra and Thomson's gazelle.

    Science.gov (United States)

    Ghanem, Y M; Fukushi, H; Ibrahim, E S M; Ohya, K; Yamaguchi, T; Kennedy, M

    2008-01-01

    Viruses related to equine herpesvirus type 1 (EHV-1) were isolated from an aborted fetus of an onager (Equus hemionus) in 1984, an aborted fetus of Grevy's zebra (Equus grevyi) in 1984 and a Thomson's gazelle (Gazella thomsoni) with nonsuppurative encephalitis in 1996, all in the USA. The mother of the onager fetus and the gazelle were kept near plains zebras (Equus burchelli). In phylogenetic trees based on the nucleotide sequences of the genes for glycoproteins B (gB), I (gI), and E (gE), and teguments including ORF8 (UL51), ORF15 (UL45), and ORF68 (US2), the onager, Grevy's zebra and gazelle isolates formed a genetic group that was different from several horse EHV-1 isolates. Within this group, the onager and gazelle isolates were closely related, while the Grevy's zebra isolate was distantly related to these two isolates. The epizootiological origin of the viruses is discussed.

  13. Removal of Cd (II) from water using the waste of jatropha fruit ( Jatropha curcas L.)

    Science.gov (United States)

    Nacke, Herbert; Gonçalves, Affonso Celso; Coelho, Gustavo Ferreira; Schwantes, Daniel; Campagnolo, Marcelo Angelo; Leismann, Eduardo Ariel Völz; Junior, Élio Conradi; Miola, Alisson Junior

    2017-10-01

    The aim of this work was to evaluate the removal of Cd (II) from water using three biosorbents originated from the biomass of jatropha (bark, endosperm, and endosperm + tegument). For that, batch tests were performed to verify the effect of solution pH, adsorbent mass, contact time, initial concentration of Cd (II), and the temperature of the process. The adsorption process was evaluated by the studies of kinetics, isotherms, and thermodynamics. The ideal conditions of solution pH were 5.5 and 8 g L-1 of adsorbent mass of biosorbents by solution volume, with an equilibrium time of 60 min. According to the Langmuir model, the maximum adsorption capacity for bark, endosperm, and bark + endosperm of jatropha was, respectively, 29.665, 19.562, and 34.674 mg g-1, predominating chemisorption in monolayers. The biosorbents presented potential for the remediation of waters contaminated with Cd (II).

  14. Effect of high energy electrons on the skin and on the underlying tissues of the rabbit. A clinical and histological study

    International Nuclear Information System (INIS)

    Legeay, G.; Vialettes, H.; Adnet, J.J.; Court, L.; Masse, R.

    1967-01-01

    The authors consider in this report the effects of high-energy electrons on rabbit teguments and on the underlying tissues after a single high dose irradiation. After briefly considering the mechanism of interaction between the electrons and matter as a function of their energy, the authors describe the dosimetry carried out, as a function of the irradiation device. The animal received surface doses of 5700 to 22100 rads in the thigh; the electron energy varied from 21 to 30 MeV. A clinical study was carried out over a period of nine months with a view to following the evolution of the damage and the functional degradation of the underlying tissues. A histological study of the induced damage was made after a second irradiation using 30 MeV electrons to produce doses of 16400 rads. Interesting observations were made concerning the damage caused to muscular and nerve tissues. (authors) [fr

  15. Partial Oculocutaneous Albinism: Two Siblings with Features of both Hermansky Pudlak and Waardenburg's Syndrome.

    Science.gov (United States)

    Ishaq, Mazhar; Niazi, Muhammad Khizar; Khan, Muhammad Saim; Nadeem, Yasser

    2015-04-01

    Albinism is an inherited abnormality of melanin synthesis with incidence of one per 20,000 births. Its clinical manifestations are related to the reduction or absence of pigmentation in the visual system and/or the skin and teguments. The clinical spectrum of Oculocutaneous Albinism (OCA) has four types ranging from OCA 1 - 4, of which OCA 1, A-1 is the most severe form. Partial cutaneous albinism which is a subtype of OCA is associated with systemic immunodeficiency disorders like Chediak Higashi (CHS), Griscelli (GS) and Hermansky-Pudlak (HPS) syndromes. A7 years boy was labeled initially as a case of Hermansky Pudlak syndrome at the age of 01 year. He as well as his 4 years old younger brother when examined in detail along with audiological investigations were diagnosed as a rare presentation of both Hermansky Pudlak and Waardenburg's syndrome.

  16. Molecular identification and characterization of prohibitin from Echinococcus granulosus.

    Science.gov (United States)

    Zhong, Xiuqin; Song, Xingju; Wang, Ning; Hu, Dandan; Liu, Tinayu; Wang, Tao; Gu, Xiaobin; Lai, Weimin; Peng, Xuerong; Yang, Guangyou

    2016-02-01

    Prohibitin (PHB) is a widely distributed protein that functions as a molecular chaperone, is involved in the regulation of cell cycle, and maintains mitochondrial structure and functions of the anti-apoptosis, senescence, and proliferation. The aim of this study was to characterize PHB in Echinococcus granulosus (EgPHB), a harmful cestode parasite of humans, many livestock species, and wild animals. We found that EgPHB is a conserved SPFH (stomatin, prohibitin, flotillin, and HflK/C) domain-containing protein, consisting of 289 amino acids, which shares 42.66-99.31% identity with PHBs from other parasites and mammals. EgPHB was located mainly in the tegument issue of protoscoleces, in the inner body of adult worms, and was expressed widely in the germinal layer. This is the first report on prohibitin from E. granulosus, and EgPHB is considered to be a valuable protein to study more in the future.

  17. Live Cell Imaging of Alphaherpes Virus Anterograde Transport and Spread

    Science.gov (United States)

    Taylor, Matthew P.; Kratchmarov, Radomir; Enquist, Lynn W.

    2013-01-01

    Advances in live cell fluorescence microscopy techniques, as well as the construction of recombinant viral strains that express fluorescent fusion proteins have enabled real-time visualization of transport and spread of alphaherpes virus infection of neurons. The utility of novel fluorescent fusion proteins to viral membrane, tegument, and capsids, in conjunction with live cell imaging, identified viral particle assemblies undergoing transport within axons. Similar tools have been successfully employed for analyses of cell-cell spread of viral particles to quantify the number and diversity of virions transmitted between cells. Importantly, the techniques of live cell imaging of anterograde transport and spread produce a wealth of information including particle transport velocities, distributions of particles, and temporal analyses of protein localization. Alongside classical viral genetic techniques, these methodologies have provided critical insights into important mechanistic questions. In this article we describe in detail the imaging methods that were developed to answer basic questions of alphaherpes virus transport and spread. PMID:23978901

  18. Bovine demodicosis: a case report Demodicose bovina: relato de caso

    Directory of Open Access Journals (Sweden)

    L.S. Lemos

    2005-12-01

    Full Text Available Tissues samples collected at necropsy from a 9 year-old Holstein cow were fixed in buffered 10% formalin for histological examination. Macroscopic finding did not reveal typical nodular cutaneous lesions. Vulvar microscopic sections of cutaneous tegument showed mites (Demodex bovis, alone and grouped in hair follicles and sebaceous glands.Amostras de tecidos colhidas à necropsia de uma vaca da raça Holandesa, nove anos de idade, foram fixadas em formol neutro a 10% e enviadas para exames histológicos. A macroscopia não revelou lesões nodulares cutâneas típicas de demodicose. Secções microscópicas de tegumento cutâneo da vulva evidenciaram ácaros (Demodex bovis isolados ou em grupos no interior de folículos pilosos e glândulas sebáceas.

  19. An autoradiographic study on the distribution of 14C-glycine in clonorchis sinensis

    International Nuclear Information System (INIS)

    Lee, S.H.; Song, C.Y.

    1977-01-01

    To study an aspect of protein metabolism in chinese liverfluke, Clonorchis sinensis, an autoradiographic study was performed. A batch of 25 ml erlenmeyer flasks, each flask containing 10 worms of C. sinensis and 10 ml of Tyrode medium with 2.5 μCi/ml of 14 C-glycine, was incubated for 1 hour in Dubnoff metabolic shaking incubator at 37 0 C. Those worms were processed for microautoradiography immediately after the incubation, and following results were obtained from the autoradiographs. The densities of black silver grains derived from 14 C-glycine were the most apparent in the subparenchymal cells, intestinal epithelium, vitelline gland cells, ovary and the wall of the seminal vesicle. Moderate grade of densities were observed in the tegument, oral sucker, pharynx, intestinal content and in the testes. The reticular tissue, ventral sucker, uterus with eggs, seminal receptacle and the content of seminal vesicle showed trace amount of silver grains. (author)

  20. Echinobothrium chisholmae n. sp. (Cestoda, Diphyllidea) from the giant shovel-nose ray Rhinobatos typus from Australia, with observations on the ultrastructure of its scolex musculature and peduncular spines.

    Science.gov (United States)

    Jones, M K; Beveridge, I

    2001-09-01

    Echinobothrium chisholmae n. sp. is described from Rhinobatos typus Bennett (Rhinobatidae), collected from Heron Island, Great Barrier Reef, Australia. E. chisholmae differs from all congeners in possessing 11 hooks in each dorsal and ventral group on the rostellum and groups of 3-6 hooklets on either side of the hooks. A single metacestode of E. chisholmae was collected from the decapod crustacean Penaeus longistylus Kubo. Yellow pigmentation of the cephalic peduncle in immature adults is caused by the accumulation of large vesicles in the distal cytoplasm of the tegument. The vesicles probably provide materials for spine formation. Ultrastructural examination of the rostellar musculature revealed that the muscles are stratified (striated-like), consisting of a periodic repetition of sarcomeres separated by perforated Z-like lines that are oblique to the long axes of the myofilaments.

  1. Herpesvirus glycoproteins undergo multiple antigenic changes before membrane fusion.

    Directory of Open Access Journals (Sweden)

    Daniel L Glauser

    Full Text Available Herpesvirus entry is a complicated process involving multiple virion glycoproteins and culminating in membrane fusion. Glycoprotein conformation changes are likely to play key roles. Studies of recombinant glycoproteins have revealed some structural features of the virion fusion machinery. However, how the virion glycoproteins change during infection remains unclear. Here using conformation-specific monoclonal antibodies we show in situ that each component of the Murid Herpesvirus-4 (MuHV-4 entry machinery--gB, gH/gL and gp150--changes in antigenicity before tegument protein release begins. Further changes then occurred upon actual membrane fusion. Thus virions revealed their final fusogenic form only in late endosomes. The substantial antigenic differences between this form and that of extracellular virions suggested that antibodies have only a limited opportunity to block virion membrane fusion.

  2. Germination, carbohydrate composition and vigor of cryopreserved Caesalpinia echinata seeds

    Directory of Open Access Journals (Sweden)

    Rafael Fonsêca Zanotti

    2012-10-01

    Full Text Available The present study investigated the germination and vigor of Caesalpinia echinata (Brazilwood seeds stored at negative temperatures. Recently harvested seeds were cryopreserved at -18º or -196ºC and periodically evaluated for germination, seed vigor and carbohydrate composition. The temperatures did not influence the germination percentages or vigor. The germination percentage decreased from 88% in recently harvested seeds to 60% after 730 days of storage. The different temperature and storage times tested did not affect the vigor seed germination as indicated by the measures of plant growth and survival. The different temperatures used did not cause changes in the carbohydrate composition. The tegument cell walls were rich in lignin, arabinose and xylose. The cytoplasm of the cotyledons and embryos had high levels of glucose, fructose, and sucrose. The cryopreservation technique here presented was effective in the conservation of Brazilwood seeds for the medium term.

  3. Adjustment of tetrazolium methodology for the evaluation of Melanoxylon brauna seeds viability

    Directory of Open Access Journals (Sweden)

    Viviana Borges Corte

    2010-09-01

    Full Text Available The objective of this work was to adjust the tetrazolium test methodology to Melanoxylon brauna seeds. The seeds werepre-conditioned by immersion in distilled water at 25ºC for 24 hours. The tegument of the seeds were removed and the embryos weresubmerged in the tetrazolium solution (pH 6.5 at concentration of 0.05, 0.1 and 0.3%, and were kept in the dark at 25ºC temperaturefor 10 and 24 hours. The embryos were evaluated for the intensity of color, presence of milky areas, aspect of the tissues, and locationof the color. The embryos were individually classified into nine viability classes. They were also submitted to the germination andseedling emergence tests. The best correlations between the standard tests of seed germination of Melanoxylon brauna were those with0.3% concentration for 10 hours. The viability is represented by classes 1, 2 and 3.

  4. Miltefosine Lipid Nanocapsules for Single Dose Oral Treatment of Schistosomiasis Mansoni: A Preclinical Study.

    Directory of Open Access Journals (Sweden)

    Maha M Eissa

    Full Text Available Miltefosine (MFS is an alkylphosphocholine used for the local treatment of cutaneous metastases of breast cancer and oral therapy of visceral leishmaniasis. Recently, the drug was reported in in vitro and preclinical studies to exert significant activity against different developmental stages of schistosomiasis mansoni, a widespread chronic neglected tropical disease (NTD. This justified MFS repurposing as a potential antischistosomal drug. However, five consecutive daily 20 mg/kg doses were needed for the treatment of schistosomiasis mansoni in mice. The present study aims at enhancing MFS efficacy to allow for a single 20mg/kg oral dose therapy using a nanotechnological approach based on lipid nanocapsules (LNCs as oral nanovectors. MFS was incorporated in LNCs both as membrane-active structural alkylphospholipid component and active antischistosomal agent. MFS-LNC formulations showed high entrapment efficiency (EE%, good colloidal properties, sustained release pattern and physical stability. Further, LNCs generally decreased MFS-induced erythrocyte hemolytic activity used as surrogate indicator of membrane activity. While MFS-free LNCs exerted no antischistosomal effect, statistically significant enhancement was observed with all MFS-LNC formulations. A maximum effect was achieved with MFS-LNCs incorporating CTAB as positive charge imparting agent or oleic acid as membrane permeabilizer. Reduction of worm load, ameliorated liver pathology and extensive damage of the worm tegument provided evidence for formulation-related efficacy enhancement. Non-compartmental analysis of pharmacokinetic data obtained in rats indicated independence of antischistosomal activity on systemic drug exposure, suggesting possible gut uptake of the stable LNCs and targeting of the fluke tegument which was verified by SEM. The study findings put forward MFS-LNCs as unique oral nanovectors combining the bioactivity of MFS and biopharmaceutical advantages of LNCs

  5. Effect of high energy electrons on the skin and on the underlying tissues of the rabbit. A clinical and histological study; Effets des electrons de haute energie sur la peau et les tissus sous-jacents du lapin. Etude clinique et histologique

    Energy Technology Data Exchange (ETDEWEB)

    Legeay, G; Vialettes, H; Adnet, J J; Court, L; Masse, R [Commissariat a l' Energie Atomique, Fontenay-aux-Roses (France). Centre d' Etudes Nucleaires

    1967-07-01

    The authors consider in this report the effects of high-energy electrons on rabbit teguments and on the underlying tissues after a single high dose irradiation. After briefly considering the mechanism of interaction between the electrons and matter as a function of their energy, the authors describe the dosimetry carried out, as a function of the irradiation device. The animal received surface doses of 5700 to 22100 rads in the thigh; the electron energy varied from 21 to 30 MeV. A clinical study was carried out over a period of nine months with a view to following the evolution of the damage and the functional degradation of the underlying tissues. A histological study of the induced damage was made after a second irradiation using 30 MeV electrons to produce doses of 16400 rads. Interesting observations were made concerning the damage caused to muscular and nerve tissues. (authors) [French] Les auteurs etudient, dans ce rapport, les effets des electrons de haute energie sur les teguments du lapin et les tissus sous-jacents apres une irradiation unique a dose elevee. Apres un rappel du mecanisme de l'interaction des electrons avec la matiere en fonction de leur energie, la dosimetrie realisee est exposee en fonction du dispositif d'irradiation. Les animaux ont recu, au niveau de la cuisse, des doses en surface de 5700 a 22100 rads; les energies des electrons vont de 21 a 30 MeV. Une etude clinique des lesions, observees pendant 9 mois, decrit leur evolution ainsi que les alterations fonctionnelles des tissus sous-jacents. Une etude histologique des lesions induites a ete realisee au cours d'une seconde experience pour des doses de 16400 rads avec des electrons de 30 MeV. D'interessantes observations ont ete faites concernant les lesions des tissus musculaires et nerveux. (auteurs)

  6. Analysis of the expression and antioxidant activity of 2-Cys peroxiredoxin protein in Fasciola gigantica.

    Science.gov (United States)

    Sangpairoj, Kant; Changklungmoa, Narin; Vanichviriyakit, Rapeepun; Sobhon, Prasert; Chaithirayanon, Kulathida

    2014-05-01

    2-Cys peroxiredoxin (Prx) is the main antioxidant enzyme in Fasciola species for detoxifying hydrogen peroxide which is generated from the hosts' immune effector cells and the parasites' own metabolism. In this study, the recombinant Prx protein from Fasciola gigantica (rFgPrx-2) was expressed and purified in a prokaryotic expression system. This recombinant protein with molecular weight of 26 kDa was enzymatically active in reduction of hydrogen peroxide both in presence of thioredoxin and glutathione systems, and also protected the supercoiled plasmid DNA from oxidative damage in metal-catalyzed oxidation (MCO) system in a concentration-dependent manner. By immunoblotting, using antibody against rFgPrx-2 as probe, a native FgPrxs, whose MW at 25 kDa, was detected in all developmental stages of the parasite. Concentrations of native FgPrxs were increasing in all stages reaching highest level in adult stage. The antibody also showed cross reactivities with corresponding proteins in some cattle helminthes. Natural antibody to FgPrxs could be detected in the sera of mice at 3 and 4 weeks after infection with F. gigantica metacercariae. By immunofluorescence, FgPrxs was highly expressed in tegument and tegumental cells, parenchyma, moderately expressed in cecal epithelial cells in early, juvenile and adult worms. Furthermore, FgPrxs was also detected in the female reproductive organs, including eggs, ovary, vitelline cells, and testis, suggesting that FgPrxs might play an essential role in protecting parasite's tissues from free radical attack during their life cycle. Thus, FgPrxs is one potential candidate for drug therapy and vaccine development. Copyright © 2014 Elsevier Inc. All rights reserved.

  7. Biochemical and immunological characterization of annexin B30 from Clonorchis sinensis excretory/secretory products.

    Science.gov (United States)

    He, Lei; Ren, Mengyu; Chen, Xueqing; Wang, Xiaoyun; Li, Shan; Lin, Jinsi; Liang, Chi; Liang, Pei; Hu, Yue; Lei, Huali; Bian, Meng; Huang, Yan; Wu, Zhongdao; Li, Xuerong; Yu, Xinbing

    2014-07-01

    Clonorchis sinensis has been classified as group I biological carcinogen for cholangiocarcinoma by the World Health Organization. Biological studies on excretory/secretory products (ESPs) enabled us to understand the pathogenesis mechanism of C. sinensis and develop new strategies for the prevention of clonorchiasis. In this study, sequence analysis showed that annexin B30 from C. sinensis (CsANXB30) is composed of four annexin repeats which were characterized by type II and III Ca(2+)-binding sites or KGD motif with the capability of Ca(2+)-binding. In addition, immunoblot assay revealed that recombinant CsANXB30 (rCsANXB30) could be recognized by the sera from rats infected with C. sinensis and the sera from rats immunized by CsESPs. Real-time PCR showed that its transcriptional level was the highest at the stage of metacercaria. Immunofluorescence assay was employed to confirm that CsANXB30 was distributed in the tegument, intestine, and egg of adult worms, as well as the tegument and vitellarium of metacercaria. rCsANXB30 was able to bind phospholipid in a Ca(2+)-dependent manner and human plasminogen in a dose-dependent manner. Moreover, cytokine and antibody measurements indicated that rats subcutaneously immunized with rCsANXB30 developed a strong IL-10 production in spleen cells and a high level of IgG1 isotype, indicating that rCsANXB30 could trigger specific humoral and cellular immune response in rats. The present results implied that CsANXB30 might be involved in a host-parasite interaction and affected the immune response of the host during C. sinensis infection.

  8. Identification and Characterization of Paramyosin from Cyst Wall of Metacercariae Implicated Protective Efficacy against Clonorchis sinensis Infection

    Science.gov (United States)

    Lv, Xiaoli; Tian, Yanli; Men, Jingtao; Zhang, Xifeng; Lei, Huali; Zhou, Chenhui; Lu, Fangli; Liang, Chi; Hu, Xuchu; Xu, Jin; Wu, Zhongdao; Li, Xuerong; Yu, Xinbing

    2012-01-01

    Human clonorchiasis has been increasingly prevalent in recent years and results in a threat to the public health in epidemic regions, motivating current strategies of vaccines to combat Clonorchis sinensis (C. sinensis). In this study, we identified C. sinensis paramyosin (CsPmy) from the cyst wall proteins of metacercariae by proteomic approaches and characterized the expressed recombinant pET-26b-CsPmy protein (101 kDa). Bioinformatics analysis indicated that full-length sequences of paramyosin are conserved in helminthes and numerous B-cell/T-cell epitopes were predicted in amino acid sequence of CsPmy. Western blot analysis showed that CsPmy was expressed at four life stages of C. sinensis, both cyst wall proteins and soluble tegumental components could be probed by anti-CsPmy serum. Moreover, immunolocalization results revealed that CsPmy was specifically localized at cyst wall and excretory bladder of metacercaria, as well as the tegument, oral sucker and vitellarium of adult worm. Both immunoblot and immunolocalization results demonstrated that CsPmy was highly expressed at the stage of adult worm, metacercariae and cercaria, which could be supported by real-time PCR analysis. Both recombinant protein and nucleic acid of CsPmy showed strong immunogenicity in rats and induced combined Th1/Th2 immune responses, which were reflected by continuous high level of antibody titers and increased level of IgG1/IgG2a subtypes in serum. In vaccine trials, comparing with control groups, both CsPmy protein and DNA vaccine exhibited protective effect with significant worm reduction rate of 54.3% (psinensis regarding its high immunogenicity and surface localization. PMID:22470461

  9. Apoptosis Governs the Elimination of Schistosoma japonicum from the Non-Permissive Host Microtus fortis

    Science.gov (United States)

    Peng, Jinbiao; Gobert, Geoffrey N.; Hong, Yang; Jiang, Weibin; Han, Hongxiao; McManus, Donald P.; Wang, Xinzhi; Liu, Jinming; Fu, Zhiqiang; Shi, Yaojun; Lin, Jiaojiao

    2011-01-01

    The reed vole, Microtus fortis, is the only known mammalian host in which schistosomes of Schistosoma japonicum are unable to mature and cause significant pathogenesis. However, little is known about how Schistosoma japonicum maturation (and, therefore, the development of schistosomiasis) is prevented in M. fortis. In the present study, the ultrastructure of 10 days post infection schistosomula from BALB/c mice and M. fortis were first compared using scanning electron microscopy and transmission electron microscopy. Electron microscopic investigations showed growth retardation and ultrastructural differences in the tegument and sub-tegumental tissues as well as in the parenchymal cells of schistosomula from M. fortis compared with those in BALB/c mice. Then, microarray analysis revealed significant differential expression between the schistosomula from the two rodents, with 3,293 down-regulated (by ≥2-fold) and 71 up-regulated (≥2 fold) genes in schistosomula from the former. The up-regulated genes included a proliferation-related gene encoding granulin (Grn) and tropomyosin. Genes that were down-regulated in schistosomula from M. fortis included apoptosis-inhibited genes encoding a baculoviral IAP repeat-containing protein (SjIAP) and cytokine-induced apoptosis inhibitor (SjCIAP), genes encoding molecules involved in insulin metabolism, long-chain fatty acid metabolism, signal transduction, the transforming growth factor (TGF) pathway, the Wnt pathway and in development. TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) and PI/Annexin V-FITC assays, caspase 3/7 activity analysis, and flow cytometry revealed that the percentages of early apoptotic and late apoptotic and/or necrotic cells, as well as the level of caspase activity, in schistosomula from M. fortis were all significantly higher than in those from BALB/c mice. PMID:21731652

  10. Low virulence potential and in vivo transformation ability in the honey bee venom treated Clinostomum complanatum.

    Science.gov (United States)

    Rehman, Abdur; Ullah, Rizwan; Jaiswal, Neeshma; Khan, M A Hannan; Rehman, Lubna; Beg, Mirza Ahmar; Malhotra, Sandeep K; Abidi, S M A

    2017-12-01

    The helminth parasites possess great capabilities to adapt themselves within their hosts and also develop strategies to render the commonly used anthelmintics ineffective leading to the development of resistance against these drugs. Besides using anthelmintics the natural products have also been tested for their anti-parasitic effects. Therapeutic efficacy of honey bee venom (HBV) has been tested in various ailments including some protozoal infections but very little is known about its anthelmintic properties. To investigate the anthelmintic effect of HBV the excysted progenetic metacercariae of Clinostomum complanatum, a heamophagic, digenetic trematode with zoonotic potential, infecting a wide variety of hosts, were obtained from Trichogaster fasciatus, a forage fish, which serves as the intermediate host. The metacercarial worms were in vitro incubated in RPMI-1640 medium containing HBV along with the controls which were devoid of HBV for the analysis of worm motility, enzyme activity, polypeptide profile and surface topographical changes. The motility of the worms was significantly reduced in a time dependent manner with an increase in the concentration of HBV. Following incubation of worms the release of cysteine proteases was inhibited in the presence of HBV as revealed by gelatine substrate gel zymography. As well as the polypeptide profile was also significantly influenced, particularly intensity/expression of M r 19.4 kDa, 24 kDa and 34 kDa was significantly reduced upon HBV treatment. The HBV treatment also inhibited antioxidant enzyme, superoxide dismutase (SOD) and Glutathione-S-transferase (GST) significantly (p < 0.05) in the worms. The scanning electron microscopy of the HBV treated worms revealed tegumental disruptions and erosion of papillae as well as spines showing vacuolation in the tegument. The HBV treated worms also showed a marked decline in the transformation rate when introduced into an experimental host which further reflect the

  11. Artyfechinostomum malayanum: Metacercariae Encysted in Pila sp. Snails Purchased from Phnom Penh, Cambodia.

    Science.gov (United States)

    Sohn, Woon-Mok; Yong, Tai-Soon; Eom, Keeseon S; Sinuon, Muth; Jeoung, Hoo-Gn; Chai, Jong-Yil

    2017-06-01

    The metacercariae of Artyfechinostomum malayanum (Leiper, 1911) Mendheim, 1943 were discovered in Pila sp. snails purchased from a market in Phnom Penh, Cambodia. They were isolated from the snails using the artificial digestion technique and were orally fed to 2 hamsters, 1 rat, and 2 mice to obtain the adult flukes. The metacercariae were round, 145-165 μm in diameter, having a cyst wall of 6-10 μm in thickness, a head collar and collar spines, and characteristic features of excretory granules. Adult flukes were recovered in the small intestines of the animals at days 14 and 32 post infection and were morphologically observed using a light microscope and a scanning electron microscope. They were plump or elongated, ventrally curved, 6.0-8.1 × 1.6-2.0 mm in size, and characterized by the head collar bearing 43 collar spines, including 5 end group ones on each side, a long cirrus sac extending beyond the posterior margin of the ventral sucker, a submedian ovary, and 2 deeply lobed testes. Eggs in uteri were operculate, ovoid to ellipsoid, and 120-135 × 68-75 μm in size. In scanning electron microscopy, the head collar was prominent with collar spines looking like horns. Scale-like tegumental spines were densely distributed on the ventral surface between the head collar and ventral sucker. Sensory papillae were distributed mainly on the tegument around suckers. By this study, it has been first confirmed that the life cycle of A. malayanum exists in Cambodia.

  12. Phytochrome controls achene germination in Bidens pilosa L. (Asteraceae by very low fluence response

    Directory of Open Access Journals (Sweden)

    Adriana Amaral-Baroli

    2001-06-01

    Full Text Available Achene without ornament of the tegument were light insensitive with germination under all tested light conditions. Achene with verrucose ornament of the tegument presented low germination under darkness and high germination under light conditions. By pre-incubation at 36° C for remotion of pre-existing Pfr and by comparison of results of counting of dark germinating achenes at the end of experiment and daily under dim green safe light (0.001mumol m-2 s-1 nm-1 we concluded that germination was controlled by phytochrome through very low fluence response.Aquênios sem ornamento do tegumento são insensíveis à luz com ocorrência de germinação sob todas as condições de luz testadas. Aquênios com ornamento verrucoso do tegumento apresentou baixa germinação sob escuro e alta germinação sob luz. A pré-incubação a 36° C para a remoção de Fve pré-existente e pela comparação dos resultados de contagem no final do experimento de aquênios que germinam no escuro e diárias sob luz verde de segurança (0.001mimol m-2s-1nm-1 concluimos que a germinação de Bidens pilosa é controlada pelo fitocromo através da resposta de fluência baixa.

  13. Human Cytomegalovirus (HCMV)-Specific CD4+ T Cells Are Polyfunctional and Can Respond to HCMV-Infected Dendritic Cells In Vitro.

    Science.gov (United States)

    Jackson, Sarah E; Sedikides, George X; Mason, Gavin M; Okecha, Georgina; Wills, Mark R

    2017-03-15

    Human cytomegalovirus (HCMV) infection and periodic reactivation are generally well controlled by the HCMV-specific T cell response in healthy people. While the CD8 + T cell response to HCMV has been extensively studied, the HCMV-specific CD4 + T cell effector response is not as well understood, especially in the context of direct interactions with HCMV-infected cells. We screened the gamma interferon (IFN-γ) and interleukin-10 (IL-10) responses to 6 HCMV peptide pools (pp65, pp71, IE1, IE2, gB, and US3, selected because they were the peptides most frequently responded to in our previous studies) in 84 donors aged 23 to 74 years. The HCMV-specific CD4 + T cell response to pp65, IE1, IE2, and gB was predominantly Th1 biased, with neither the loss nor the accumulation of these responses occurring with increasing age. A larger proportion of donors produced an IL-10 response to pp71 and US3, but the IFN-γ response was still dominant. CD4 + T cells specific to the HCMV proteins studied were predominantly effector memory cells and produced both cytotoxic (CD107a expression) and cytokine (macrophage inflammatory protein 1β secretion) effector responses. Importantly, when we measured the CD4 + T cell response to cytomegalovirus (CMV)-infected dendritic cells in vitro , we observed that the CD4 + T cells produced a range of cytotoxic and secretory effector functions, despite the presence of CMV-encoded immune evasion molecules. CD4 + T cell responses to HCMV-infected dendritic cells were sufficient to control the dissemination of virus in an in vitro assay. Together, the results show that HCMV-specific CD4 + T cell responses, even those from elderly individuals, are highly functional and are directly antiviral. IMPORTANCE Human cytomegalovirus (HCMV) infection is carried for a lifetime and in healthy people is kept under control by the immune system. HCMV has evolved many mechanisms to evade the immune response, possibly explaining why the virus is never eliminated

  14. On astronomical drawing [1846

    Science.gov (United States)

    Smyth, Charles Piazzi

    Reprinted from the Memoirs of the Royal Astronomical Society 15, 1846, pp. 71-82. With annotations and illustrations added by Klaus Hentschel. The activities of the Astronomer Royal for Scotland, Charles Piazzi Smyth (1819-1900), include the triangulation of South African districts, landscape painting, day-to-day or tourist sketching, the engraving and lithographing of prominent architectural sites, the documentary photography of the Egyptian pyramids or the Tenerife Dragon tree, and `instant photographs' of the clouds above his retirement home in Clova, Ripon. His colorful records of the aurora polaris, and solar and terrestrial spectra all profited from his trained eye and his subtle mastery of the pen and the brush. As his paper on astronomical drawing, which we chose to reproduce in this volume, amply demonstrates, he was conversant in most of the print technology repertoire that the 19th century had to offer, and carefully selected the one most appropriate to each sujet. For instance, he chose mezzotint for the plates illustrating Maclear's observations of Halley's comet in 1835/36, so as to achieve a ``rich profundity of shadows, the deep obscurity of which is admirably adapted to reproduce those fine effects of chiaroscuro frequently found in works where the quantity of dark greatly predominates.'' The same expertise with which he tried to emulate Rembrandt's chiaroscuro effects he applied to assessing William and John Herschel's illustrations of nebulae, which appeared in print between 1811 and 1834. William Herschel's positive engraving, made partly by stippling and partly by a coarse mezzotint, receives sharp admonishment because of the visible ruled crossed lines in the background and the fact that ``the objects, which are also generally too light, [have] a much better definition than they really possess.'' On the other hand, John Herschel's illustration of nebulae and star clusters, given in negative, ``in which the lights are the darkest part of the

  15. Installation of groundwater observation tubes OL-PVP36-38 and drilling of shallow drillholes OL-PP70-71 at Olkiluoto in Eurajoki 2011

    International Nuclear Information System (INIS)

    Toropainen, V.

    2012-05-01

    In order to widen the groundwater monitoring network at Olkiluoto, Posiva Oy contracted Suomen Malmi Oy (Smoy) to install new groundwater observation tubes to three locations and to drill two shallow drillholes with standpipes. The identification numbers of the groundwater observation tubes are OL-PVP36, OL-PVP37A, 37B, 37C, OL-PVP38A, 38B, 38C and 38D, and the shallow drillholes are named OL-PP70 and OL-PP71. The observation tubes were installed and the shallow holes drilled between September 22nd and October 12th in 2011. The drilling rig used in the installation work was a GM-200 rig. Drilling equipment consisted of casing tubes (90/77 mm) with drilling bit, 55 mm geo rods and 64 mm drilling bits and T76-equipment for drilling the shallow holes. Monitoring pipes (PVC, 60/52 mm) were lowered into the holes inside the casings. The monitoring pipes consist of a lower section of riser pipe, a middle section of screen pipe and an upper section of riser pipe. The screen pipe slot size is 0.3 mm and the length of the screen section is two metres. Protective stainless steel covers with lock-up caps were installed around the monitoring tubes and the shallow drillholes. In addition to the installation of the tubes, the work included water level measurements after installation. The core samples of the shallow drillholes were logged and reported by geologist. Geological logging included the following parameters: lithology, foliation, fracture parameters, fractured zones, core loss, weathering, fracture frequency, RQD and rock quality. (orig.)

  16. Installation of groundwater observation tubes OL-PVP36-38 and drilling of shallow drillholes OL-PP70-71 at Olkiluoto in Eurajoki 2011

    Energy Technology Data Exchange (ETDEWEB)

    Toropainen, V. [Suomen Malmi Oy, Espoo (Finland)

    2012-05-15

    In order to widen the groundwater monitoring network at Olkiluoto, Posiva Oy contracted Suomen Malmi Oy (Smoy) to install new groundwater observation tubes to three locations and to drill two shallow drillholes with standpipes. The identification numbers of the groundwater observation tubes are OL-PVP36, OL-PVP37A, 37B, 37C, OL-PVP38A, 38B, 38C and 38D, and the shallow drillholes are named OL-PP70 and OL-PP71. The observation tubes were installed and the shallow holes drilled between September 22nd and October 12th in 2011. The drilling rig used in the installation work was a GM-200 rig. Drilling equipment consisted of casing tubes (90/77 mm) with drilling bit, 55 mm geo rods and 64 mm drilling bits and T76-equipment for drilling the shallow holes. Monitoring pipes (PVC, 60/52 mm) were lowered into the holes inside the casings. The monitoring pipes consist of a lower section of riser pipe, a middle section of screen pipe and an upper section of riser pipe. The screen pipe slot size is 0.3 mm and the length of the screen section is two metres. Protective stainless steel covers with lock-up caps were installed around the monitoring tubes and the shallow drillholes. In addition to the installation of the tubes, the work included water level measurements after installation. The core samples of the shallow drillholes were logged and reported by geologist. Geological logging included the following parameters: lithology, foliation, fracture parameters, fractured zones, core loss, weathering, fracture frequency, RQD and rock quality. (orig.)

  17. Human IgG1 Responses to Surface Localised Schistosoma mansoni Ly6 Family Members Drop following Praziquantel Treatment.

    Directory of Open Access Journals (Sweden)

    Iain W Chalmers

    Full Text Available The heptalaminate-covered, syncytial tegument is an important anatomical adaptation that enables schistosome parasites to maintain long-term, intravascular residence in definitive hosts. Investigation of the proteins present in this surface layer and the immune responses elicited by them during infection is crucial to our understanding of host/parasite interactions. Recent studies have revealed a number of novel tegumental surface proteins including three (SmCD59a, SmCD59b and Sm29 containing uPAR/Ly6 domains (renamed SmLy6A SmLy6B and SmLy6D in this study. While vaccination with SmLy6A (SmCD59a and SmLy6D (Sm29 induces protective immunity in experimental models, human immunoglobulin responses to representative SmLy6 family members have yet to be thoroughly explored.Using a PSI-BLAST-based search, we present a comprehensive reanalysis of the Schistosoma mansoni Ly6 family (SmLy6A-K. Our examination extends the number of members to eleven (including three novel proteins and provides strong evidence that the previously identified vaccine candidate Sm29 (renamed SmLy6D is a unique double uPAR/Ly6 domain-containing representative. Presence of canonical cysteine residues, signal peptides and GPI-anchor sites strongly suggest that all SmLy6 proteins are cell surface-bound. To provide evidence that SmLy6 members are immunogenic in human populations, we report IgG1 (as well as IgG4 and IgE responses against two surface-bound representatives (SmLy6A and SmLy6B within a cohort of S. mansoni-infected Ugandan males before and after praziquantel treatment. While pre-treatment IgG1 prevalence for SmLy6A and SmLy6B differs amongst the studied population (7.4% and 25.3% of the cohort, respectively, these values are both higher than IgG1 prevalence (2.7% for a sub-surface tegumental antigen, SmTAL1. Further, post-treatment IgG1 levels against surface-associated SmLy6A and SmLy6B significantly drop (p = 0.020 and p < 0.001, respectively when compared to rising Ig

  18. Caracterização da microbiota por fungos filamentosos no tegumento hígido de bovinos de corte Characterization of filamentous fungal flora from the integument of healthy cattle

    Directory of Open Access Journals (Sweden)

    Christiane Duarte Pombo do Amaral

    2011-12-01

    Full Text Available O tegumento de bovinos pode albergar uma grande diversidade de fungos filamentosos, potencialmente capazes de causar dermatoses, incluindo fungos do grupo dos dermatófitos. O presente estudo teve por objetivos caracterizar a microbiota de fungos filamentosos do tegumento hígido de bovinos de corte, verificar a presença de dermatófitos como parte integrante da microbiota, assim como avaliar a ocorrência de dermatófitos no solo dos estabelecimentos estudados. Durante o período de um ano, 56 bovinos sadios de quatro propriedades da fronteira Oeste do RS foram submetidos a coletas mensais de pelos e raspado cutâneo. No mesmo período, foram obtidas amostras de solo para pesquisa de dermatófitos. As análises dos resultados indicaram uma microbiota de fungos filamentosos classificada em 30 gêneros, dos quais os mais frequentes foram (nesta ordem: Nigrospora, Fusarium, Curvularia, Alternaria, Epicoccum, Paecilomyces e Trichoderma. Nas amostras de pelame, obteve-se o isolamento de Trichophyton mentagrophytes (0,4%, Microsporum gypseum (0,3% e Trichophyton verrucosum (0,1%. M. gypseum foi o dermatófito mais frequentemente isolado do solo. Os resultados obtidos evidenciaram que a microbiota fúngica do tegumento hígido de bovinos de corte comporta-se de maneira transitória e que bovinos sadios não atuam como portadores assintomáticos de T. verrucosum, sugerindo que esta espécie de dermatófito é não residente no tegumento dos animais.The tegument of cattle may harbor a great diversity of filamentous fungi that can potentially cause skin diseases, including fungi belonging to the dermatophyte group. This study aimed to characterize the population of filamentous fungi from a healthy coat of cattle, checking for dermatophytes as part of the microbiota, as well as evaluating the presence of dermatophytes in the soil occupied by the animals. During the period of one year, 56 healthy cattle in four properties in the western boundary of the

  19. Tetrazolium test in the evaluation of watermelon physiological seed quality/ Teste de tetrazólio para avaliação da qualidade fisiológica de sementes de melancia

    Directory of Open Access Journals (Sweden)

    Luciana Magda de Oliveira

    2007-08-01

    Full Text Available The tetrazolium test is one of the most promising methods to estimate in a fast way viability and vigor of seeds. Its usage in watermelon seeds requires some care with the imbibition and handling due to presence of mucilaginous layer adherent to the tegument. The objective of this research work was to define the imbibition time and the method of scarification for removing the mucilage, during pre – conditioning of the tetrazolium test in watermelon. Seeds from different watermellon cultivars were immersed in water at 30°C from 12 to 18 hours, and sloughed in cal, fine and thick sand. After longitudinal cut in the distal portion to the embryonic axis, the tegument was removed and embryos were kept in water for additonal 2 hours at 30°C for hand removal of the remaining membrane. After definition of the ideal pre – conditioning method, the embryos were immersed in tetrazolium solution in concentrations of 0,075%; 0, 5% and 1% for 3 and 4 hours at 30°C. The immersion of watermelon seeds in water at 30°C for 12 hours, mucilage removal with fine sand, longitudinal cut in the distal portion to the embryonic axis, and removal of tegument, followed by the permanence of embryos in water for 2 hours at 30°C for the hand removal of the internal membrane and immersion in the tetrazolium solution at 0,075% for 4 hours at 30°C are adequate procedures for the evaluation of watermelon seed quality.O teste de tetrazólio é um dos métodos mais promissores para estimar, de forma rápida, a viabilidade e o vigor das sementes. Sua utilização em sementes de melancia requer cuidados na embebição e manuseio pela presença de camada mucilaginosa aderente ao tegumento. O objetivo do trabalho foi definir o tempo de embebição e do método de escarificação para retirada da mucilagem, no pré-condicionamento do teste de tetrazólio, sementes de diferentes cultivares de melancia. As sementes foram imersas em água a 30oC por 12 e 18 horas, e escarificadas

  20. Germinação in vitro de sementes de alcachofra In vitro artichoke seed germination

    Directory of Open Access Journals (Sweden)

    Cassieli F de Moraes

    2010-03-01

    Full Text Available A baixa taxa de multiplicação e alta de contaminação dos explantes são algumas das dificuldades na micropropagação da alcachofra. A germinação de sementes in vitro pode ser uma alternativa de obtenção de explantes sadios para estabelecimento de futuros cultivos in vitro. O trabalho desenvolvido no Laboratório de Biotecnologia Vegetal da UPF-FAMV teve por objetivo avaliar a germinação in vitro de sementes de alcachofra cv. Nobre, em três experimentos, testando concentrações de cloro ativo na assepsia das sementes; tratamentos do tegumento (mantido intacto, com cortes laterais ou eliminação; condições de luminosidade (claro ou escuro; e dois meios de cultura [meio MS, com concentração de sais reduzida à metade (M1 e meio MS completo(M2]. Em ambos foram adicionados 30 g L-1 de sacarose e 7 g L-1 de ágar, sendo o pH ajustado para 5,6 ± 0,1 com NaOH. Os cultivos foram realizados em câmara de crescimento. A obtenção de plântulas sadias de alcachofra em curto espaço de tempo (sete dias, para utilização como fonte de explantes é viável a partir da germinação in vitro de sementes sem o tegumento (77,5% de germinação, utilizando os meios de cultura M1 ou M2 e câmara de crescimento desprovida de luz. Nestas condições, a assepsia das sementes pode ser realizada com álcool 70% por 30 min e posterior imersão em solução contendo 2% de cloro ativo por dez minutos, antes da remoção do tegumento.Low multiplication rates and high contamination in the explants are some of the difficulties in artichoke micropropagation. In vitro seed germination may be an alternative to obtain healthy explants for use in future in vitro cultivation. This project developed at the laboratory of Universidade de Passo Fundo was established to evaluate cv. 'Nobre' artichoke seeds in vitro germination. In three experiments, active chloride concentrations on seed aseptic technique; tegument treatment (kept intact, with side cuts and

  1. Formação do porta-enxerto Trifoliata: época de semeadura e tegumento na emergência de plântulas Production of Trifoliata rootstock: season and seedcoat in seedling emergency

    Directory of Open Access Journals (Sweden)

    Roberto Pedroso de Oliveira

    2007-02-01

    Full Text Available O objetivo deste trabalho foi avaliar o efeito da época de semeadura e do tegumento da semente na emergência de plântulas e o tempo necessário para formação do porta-enxerto Trifoliata. O experimento foi realizado em casa de vegetação, em Pelotas-RS. A semeadura foi realizada em tubetes de 50cm3, contendo substrato comercial. Os tratamentos compreenderam duas épocas de semeadura (inverno e primavera e três tratamentos do tegumento das sementes (tegumento íntegro, com furo no endosperma, e sem tegumento. Esses foram dispostos em delineamento de blocos ao acaso, em esquema fatorial, com quatro repetições, sendo as unidades experimentais constituídas por 42 tubetes com uma semente cada. Os porta-enxertos foram considerados aptos para o transplantio quando atingiram 15cm de altura. A velocidade e a porcentagem final de germinação de Trifoliata foi significativamente superior na primavera (95,6% em relação ao inverno (83,6%. Independentemente da época do ano, a remoção do tegumento favoreceu a germinação (96,9% em relação ao uso de semente com tegumento íntegro (84,8%. O período necessário para a formação dos porta-enxertos nos tubetes foi de 70 dias na primavera e de 130 dias no inverno.This study was aimed at evaluating the effect of the time of sowing and the seedcoat in seedling emergency and production of Trifoliata rootstock. The experiment was carried out in a glasshouse, in Pelotas-RS. Sowing was performed in dibble tubes of 50cm3, containing commercial substrate. Two sowing plates (winter and spring and three seedcoat treatments (integral tegument, with a hole in the endosperm and without tegument were studied. The experimental design was a randomized complete block, in a factorial schedule, with four replications and experimental units composed by 42 dibble tubes with one seed each. The rootstocks were considered adequate for the transplant when they reached 15cm height. The speed and final germination

  2. Bacterial ecology of tilapia fresh fillets and some factors that can influence their microbial quality Ecologia bacteriana de filés frescos de tilápia e de pontos capazes de influenciar a sua qualidade microbiológica

    Directory of Open Access Journals (Sweden)

    Cleube Andrade Boari

    2008-12-01

    Full Text Available The purpose of research was to investigate the bacterial ecology of tilapia (Oreochromis niloticus fresh fillets and some factors that can influence its microbial quality. Samples of fish cultivation water (n = 20, tilapia tegument and gut (n = 20 and fresh fillets (n = 20 were collected in an experimental tilapia aquaculture located in the city of Lavras, Minas Gerais, Brazil. Staphylococcus spp., Aeromonas spp., Enterococcus spp. and Enterobacteriaceae were quantified using selective plating. For the enumeration of Pseudomonas spp., the most probable number technique (MPN was utilized. Bacterial colonies (n = 198 were identified by Gram strain and biochemical tests. Aeromonas spp., Pseudomonas spp., Enterococcus spp. and Enterobacteriaceae were found in the cultivation water (water from a fishpond cultivation, tegument, gut, and fresh fillets. Staphylococcus spp. was not isolated in the cultivation water. Salmonella spp. was not detected. The count variable of 10 to 10³ CFU or MPN.(g or mL-1. Associated to freshwater tilapia fillet processing, there is a large variety of microorganisms related to foodborne illnesses and fish products deterioration.A presente pesquisa foi conduzida com o objetivo de se investigar a ecologia microbiana de filés frescos de tilápia (Oreochromis niloticus e de pontos capazes de influenciar em sua qualidade microbiológica. Para sua condução, amostras de água de cultivo (n = 20, tegumento e trato gastrintestinal (n = 20 e filés frescos (n = 20 foram coletados em um sistema de produção aqüícola de água doce localizado em Lavras, Minas Gerais, Brasil. Staphylococcus spp., Aeromonas spp., Enterococcus spp., Enterobacteriaceae foram quantificados através de plaqueamento seletivo. Pseudomonas spp. foi enumerada através da técnica do número mais provável. Colônias (n = 198 foram identificadas por coloração de Gram e testes bioquímicos. Aeromonas spp., Pseudomonas spp., Enterococcus spp. e

  3. Insights into the biological features of the antigenic determinants recognized by four monoclonal antibodies in redia and adult stages of the liver fluke Fasciola hepatica.

    Science.gov (United States)

    Alba, Annia; Sánchez, Jorge; Hernández, Hilda; Mosqueda, Maryani; Rodríguez, Suanel Y; Capó, Virginia; Otero, Oscar; Alfonso, Carlos; Marcet, Ricardo; Sarracent, Jorge

    2016-09-01

    Fasciola hepatica is a digenean trematode which infects a wide variety of domestic animals and also humans. Previous studies have demonstrated that four monoclonal antibodies (Mabs) against the total extract of F. hepatica redia (named as 1E4, 6G11, 4E5 and 4G11) also recognized the excretion - secretion antigens (ES Ag) of adult parasites, which is a biologically-relevant mixture of molecules with functional roles during infection and immune evasion on definitive hosts. In the present report we describe the partial characterization of the epitopes recognized by these Mabs by heat treatment, mercaptoethanol reduction, pronase proteolysis and sodium peryodate oxidation, which suggested their predominant protein and conformational nature. Also, a comparative study using immunodetection assays on crude extracts and on histological sections of both rediae and adults of F. hepatica were performed to explore the expression pattern of the antigenic determinants in these developmental stages. From these experiments it was found that the Mabs reacted most likely with the same proteins of approximately 64 and 105 kDa present on both rediae and adult's extracts. However, the 1E4, 6G11 and 4E5 Mabs also recognized other molecules of the total extract of F. hepatica adults, a fact that constitutes an evidence of the antigenic variation between both stages and points at a certain biological relevance of the recognized antigenic determinants. Immunolocalization studies on histological sections revealed that all Mabs reacted with the tegument of F. hepatica in both rediae and adults stages, while the epitopes recognized by 1E4, 6G11 and 4E5 antibodies were also preferentially localized in the intestinal caeca and in different organs of the reproductive system of adult specimens. The immunogenicity of these antigenic determinants, their conserved status among different stages of the life cycle of F. hepatica and their presence in both tegument and ES Ag of adult parasites

  4. Molecular insights into a tetraspanin in the hydatid tapeworm Echinococcus granulosus.

    Science.gov (United States)

    Hu, Dandan; Song, Xingju; Xie, Yue; Zhong, Xiuqin; Wang, Ning; Zheng, Yu; Gu, Xiaobin; Wang, Tao; Peng, Xuerong; Yang, Guangyou

    2015-06-10

    Cystic echinococcosis (hydatid disease), caused by the tapeworm Echinococcus granulosus (class Cestoda; family Taeniidae), is a neglected tropical disease that results in morbidity and mortality in millions of humans, as well as in huge economic losses in the livestock industry globally. Proteins from the tetraspanin family in parasites have recently become regarded as crucial molecules in interaction with hosts in parasitism and are therefore suitable for the development of vaccines and diagnostic agents. However, no information is available to date on E. granulosus tetraspanin. In this study, a uroplakin-I-like tetraspanin (Eg-TSP1) of E. granulosus was cloned and expressed in E. coli. The immunolocalization of Eg-TSP1 in different life stages of E. granulosus was determined using specific polyclonal antibody. The antibody and cytokine profiles of mice that immunized with recombinant Eg-TSP1 (rEg-TSP1) were measured for the immunogenicity analysis of this protein. Additionally, we use RNA interference method to explore the biological function of Eg-TSP1 in larva of E. granulosus. Immunofluorescence analysis showed that endogenous Eg-TSP1 mainly localized in the tegument of larvae and adults. Significantly elevated levels of antibodies IgG1 and IgG2a and of cytokines IFN-γ and IL-12 were observed in the sera of mice after immunization with rEg-TSP1, suggesting a typical T helper (Th)1-mediated immune response elicited by rEg-TSP1. On further probing the role of Eg-TSP1 in E. granulosus by RNA interference, we found that a thinner tegmental distal cytoplasm was induced in protoscoleces treated with siRNA-132 compared to controls. This is the first report characterizing a tetraspanin from the tapeworm E. granulosus. Our results suggest that Eg-TSP1 is associated with biogenesis of the tegument and maintenance of structural integrity of E. granulosus and could therefore be a candidate intervention target for control of hydatid disease.

  5. Molecular cloning and characterization of Fasciola gigantica thioredoxin-glutathione reductase.

    Science.gov (United States)

    Changklungmoa, Narin; Kueakhai, Pornanan; Sangpairoj, Kant; Chaichanasak, Pannigan; Jaikua, Wipaphorn; Riengrojpitak, Suda; Sobhon, Prasert; Chaithirayanon, Kulathida

    2015-06-01

    The Fasciola gigantica thioredoxin-glutathione reductase (FgTGR) gene is a fusion between thioredoxin reductase (TR) and a glutaredoxin (Grx) gene. FgTGR was cloned by polymerase chain reaction (PCR) from adult complementary DNA (cDNA), and its sequences showed two isoforms, i.e., the cytosolic and mitochondrial FgTGR. Cytosolic FgTGR (cytFgTGR) was composed of 2370 bp, and its peptide had no signal sequence and hence was not a secreted protein. Mitochondrial FgTGR (mitFgTGR) was composed of 2506 bp with a signal peptide of 43 amino acids; therefore, it was a secreted protein. The putative cytFgTGR and mitFgTGR peptides comprised of 598 and 641 amino acids, respectively, with a molecular weight of 65.8 kDa for cytFgTGR and mitFgTGR, with a conserved sequence (CPYC) of TR, and ACUG and CVNVGC of Grx domains. The recombinant FgTGR (rFgTGR) was expressed in Escherichia coli BL21 (DE3) and used for production for a polyclonal antibody in rabbits (anti-rFgTGR). The FgTGR protein expression, estimated by indirect ELISA using the rabbit anti-rFgTGR as probe, showed high levels of expression in eggs, and 2- and 4-week-old juveniles and adults. The rFgTGR exhibited specific activities in the 5,5'-dithiobis (2-nitro-benzoic acid) (DTNB) reductase assay for TR activity and in β-hydroxyethul disulfide (HED) for Grx activity. When analyzed by immunoblotting and immunohistochemistry, rabbit anti-rFgTGR reacted with natural FgTGR at a molecular weight of 66 kDa from eggs, whole body fraction (WB) of metacercariae, NEJ, 2- and 4-week-old juveniles and adults, and the tegumental antigen (TA) of adult. The FgTGR protein was expressed at high levels in the tegument of 2- and 4-week-old juveniles. The FgTGR may be one of the major factors acting against oxidative stresses that can damage the parasite; hence, it could be considered as a novel vaccine or a drug target.

  6. Identification and characterization of paramyosin from cyst wall of metacercariae implicated protective efficacy against Clonorchis sinensis infection.

    Directory of Open Access Journals (Sweden)

    Xiaoyun Wang

    Full Text Available Human clonorchiasis has been increasingly prevalent in recent years and results in a threat to the public health in epidemic regions, motivating current strategies of vaccines to combat Clonorchis sinensis (C. sinensis. In this study, we identified C. sinensis paramyosin (CsPmy from the cyst wall proteins of metacercariae by proteomic approaches and characterized the expressed recombinant pET-26b-CsPmy protein (101 kDa. Bioinformatics analysis indicated that full-length sequences of paramyosin are conserved in helminthes and numerous B-cell/T-cell epitopes were predicted in amino acid sequence of CsPmy. Western blot analysis showed that CsPmy was expressed at four life stages of C. sinensis, both cyst wall proteins and soluble tegumental components could be probed by anti-CsPmy serum. Moreover, immunolocalization results revealed that CsPmy was specifically localized at cyst wall and excretory bladder of metacercaria, as well as the tegument, oral sucker and vitellarium of adult worm. Both immunoblot and immunolocalization results demonstrated that CsPmy was highly expressed at the stage of adult worm, metacercariae and cercaria, which could be supported by real-time PCR analysis. Both recombinant protein and nucleic acid of CsPmy showed strong immunogenicity in rats and induced combined Th1/Th2 immune responses, which were reflected by continuous high level of antibody titers and increased level of IgG1/IgG2a subtypes in serum. In vaccine trials, comparing with control groups, both CsPmy protein and DNA vaccine exhibited protective effect with significant worm reduction rate of 54.3% (p<0.05 and 36.1% (p<0.05, respectively. In consistence with immune responses in sera, elevated level of cytokines IFN-γ and IL-4 in splenocytes suggested that CsPmy could induce combined cellular immunity and humoral immunity in host. Taken together, CsPmy could be a promising vaccine candidate in the prevention of C. sinensis regarding its high immunogenicity

  7. Identification and characterization of a member of Rab subfamily, Rab8, from Clonorchis sinensis.

    Science.gov (United States)

    Liang, Pei; He, Lei; Yu, Jinyun; Xie, Zhizhi; Chen, Xueqing; Mao, Qiang; Liang, Chi; Huang, Yan; Lu, Gang; Yu, Xinbing

    2015-05-01

    The Rabs act as a binary molecular switch that utilizes the conformational changes associated with the GTP/GDP cycle to elicit responses from target proteins. It regulates a broad spectrum of cellular processes including cell proliferation, cytoskeletal assembly, and intracellular membrane trafficking in eukaryotes. The Rab8 from Clonorchis sinensis (CsRab8) was composed of 199 amino acids. The deduced amino acid sequence shared above 50% identities with other species from trematode, tapeworm, mammal, insecta, nematode, and reptile, respectively. The homologous analysis of sequences showed the conservative domains: G1 box (GDSGVGKS), G2 box (T), G3 box (DTAG), G4 box (GNKCDL), and G5 box. In addition, the structure modeling had also shown other functional domains: GTP/Mg(2+) binding sites, switch I region, and switch II region. A phylogenic tree analysis indicated that the CsRab8 was clustered with the Rab from Schistosoma japonicum, and trematode and tapeworm came from the same branch, which was different from an evolutional branch built by other species, such as mammal animal, insecta, nematode, and reptile. The recombinant CsRab8 protein was expressed in Escherichia coli and the purified protein was a soluble molecule by 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis. CsRab8 was identified as a component of excretory/secretory products of C. sinensis by western blot analysis. The transcriptional level of CsRab8 at metacercaria stage was the highest at the four stages and higher by 56.49-folds than that at adult worm, 1.23-folds than that at excysted metacercaria, and 2.69-folds than that at egg stage. Immunohistochemical localization analysis showed that CsRab8 was specifically distributed in the tegument, vitellarium, eggs, and testicle of adult worms, and detected on the vitellarium and tegument of metacercaria. Combined with the results, CsRab8 is indispensable for survival and development of parasites, especially for regulating

  8. Cardamonin, a schistosomicidal chalcone from Piper aduncum L. (Piperaceae) that inhibits Schistosoma mansoni ATP diphosphohydrolase.

    Science.gov (United States)

    de Castro, Clarissa C B; Costa, Poliana S; Laktin, Gisele T; de Carvalho, Paulo H D; Geraldo, Reinaldo B; de Moraes, Josué; Pinto, Pedro L S; Couri, Mara R C; Pinto, Priscila de F; Da Silva Filho, Ademar A

    2015-09-15

    Schistosomiasis is one of the world's major public health problems, and praziquantel (PZQ) is the only available drug to treat this neglected disease with an urgent demand for new drugs. Recent studies indicated that extracts from Piper aduncum L. (Piperaceae) are active against adult worms of Schistosoma mansoni, the major etiological agent of human schistosomiasis. We investigated the in vitro schistosomicidal activity of cardamonin, a chalcone isolated from the crude extract of P. aduncum. Also, this present work describes, for the first time, the S. mansoni ATP diphosphohydrolase inhibitory activity of cardamonin, as well as, its molecular docking with S. mansoni ATPDase1, in order to investigate its mode of inhibition. In vitro schistosomicidal assays and confocal laser scanning microscopy were used to evaluate the effects of cardamonin on adult schistosomes. Cell viability was measured by MTT assay, and the S. mansoni ATPase activity was determined spectrophotometrically. Identification of the cardamonin binding site and its interactions on S. mansoni ATPDase1 were made by molecular docking experiments. A bioguided fractionation of the crude extract of P. aduncum was carried out, leading to identification of cardamonin as the active compound, along with pinocembrin and uvangoletin. Cardamonin (25, 50, and 100 µM) caused 100% mortality, tegumental alterations, and reduction of oviposition and motor activity of all adult worms of S. mansoni, without affecting mammalian cells. Confocal laser scanning microscopy showed tegumental morphological alterations and changes on the numbers of tubercles of S. mansoni worms in a dose-dependent manner. Cardamonin also inhibited S. mansoni ATP diphosphohydrolase (IC50 of 23.54 µM). Molecular docking studies revealed that cardamonin interacts with the Nucleotide-Binding of SmATPDase 1. The nature of SmATPDase 1-cardamonin interactions is mainly hydrophobic and hydrogen bonding. This report provides evidence for the in vitro

  9. Effects of Triclabendazole and Al bendazole on treatment of Fascioliasis

    International Nuclear Information System (INIS)

    Mawoad, M.A.F.; Amin, M.M.; Elnashar, D. E.

    2010-01-01

    decrease in liver damage. On the other hand, scanning electron microscopy of whole worm revealed extensive swelling and blebbing of the tegument on ventral surface. Some tegumental loss had occurred in the oral region of the fluke. The oral sucker of Fasciola hepatica became wide and the ventral sucker became also wide and eroded. The obtained results revealed that the TCBZ and ABZ are both effective in getting rid of Fasciola hepatica infection. However, TCBZ tended to has more anthelmintic effect than ABZ. Additionally, TCBZ has an ameliorative effect on liver, it also decreased liver fluke egg counts more than that induced by ABZ

  10. The diterpenoid 7-keto-sempervirol, derived from Lycium chinense, displays anthelmintic activity against both Schistosoma mansoni and Fasciola hepatica.

    Directory of Open Access Journals (Sweden)

    Jennifer Edwards

    2015-03-01

    Full Text Available BACKGROUND: Two platyhelminths of biomedical and commercial significance are Schistosoma mansoni (blood fluke and Fasciola hepatica (liver fluke. These related trematodes are responsible for the chronic neglected tropical diseases schistosomiasis and fascioliasis, respectively. As no vaccine is currently available for anti-flukicidal immunoprophylaxis, current treatment is mediated by mono-chemical chemotherapy in the form of mass drug administration (MDA (praziquantel for schistosomiasis or drenching (triclabendazole for fascioliasis programmes. This overreliance on single chemotherapeutic classes has dramatically limited the number of novel chemical entities entering anthelmintic drug discovery pipelines, raising significant concerns for the future of sustainable blood and liver fluke control. METHODOLOGY/ PRINCIPLE FINDINGS: Here we demonstrate that 7-keto-sempervirol, a diterpenoid isolated from Lycium chinense, has dual anthelmintic activity against related S. mansoni and F. hepatica trematodes. Using a microtiter plate-based helminth fluorescent bioassay (HFB, this activity is specific (Therapeutic index = 4.2, when compared to HepG2 cell lines and moderately potent (LD50 = 19.1 μM against S. mansoni schistosomula cultured in vitro. This anti-schistosomula effect translates into activity against both adult male and female schistosomes cultured in vitro where 7-keto-sempervirol negatively affects motility/behaviour, surface architecture (inducing tegumental holes, tubercle swelling and spine loss/shortening, oviposition rates and egg morphology. As assessed by the HFB and microscopic phenotypic scoring matrices, 7-keto-sempervirol also effectively kills in vitro cultured F. hepatica newly excysted juveniles (NEJs, LD50 = 17.7 μM. Scanning electron microscopy (SEM evaluation of adult F. hepatica liver flukes co-cultured in vitro with 7-keto-sempervirol additionally demonstrates phenotypic abnormalities including breaches in tegumental

  11. Sodium exchange process in lower vertebrates studies by means of {sup 24}Na. Their interpretation as a 'purification' of radioactive water; Les echanges de sodium des vertebres inferieurs etudies a l'aide du {sup 24}Na. Leur interpretation comme une 'epuration' d'eau radioactive

    Energy Technology Data Exchange (ETDEWEB)

    Maetz, J [Commissariat a l' Energie Atomique, Saclay (France). Centre d' Etudes Nucleaires

    1958-07-01

    Fresh-water fish and amphibians live in an aquatic medium which is very hypotonic with respect to their internal medium. By osmosis the water passes through their permeable teguments (gills, mucous membranes, skin). It is known from the work of Krogh (1937-1939) that this water is eliminated by the kidney, which emits a copious diluted urine. Yet however great the dilution, the urinary excretion results in a loss of salts for the organism. This loss is compensated, even in the absence of any nourishment, by active absorption of ions. Some recent workers have attempted, by using radioactive isotopes, to establish the balance between the input and output flows of ions through the permeable membranes in vivo, whereas chemical analytical methods only allow the net result of these exchanges to be obtained. We have attempted to present here a new method for the simultaneous measurement of the sodium flow takes over the whole animal. This method has been applied to the fresh-water fish Carassius auratus and to the frog Rana esculenta. In the course of this investigation we have considered interpreting it as a 'purification of radioactive water'. (author) [French] Les poissons d'eau douce et les amphibiens vivent dans un milieu aquatique tres hypotonique par rapport a leur milieu interieur. Par osmose, l'eau penetre a travers leurs teguments permeables (branchies, muqueuses, peau). On sait depuis les travaux de Krogh (1937-1939) que cette eau est eliminee par le rein qui emet une urine abondante et diluee. Mais si diluee soit-elle, l'excretion uranaire entraine une perte de sels pour l'organisme. Cette perte est compensee, meme en l'absence de toute alimentation, par une absorption active d'ions. Quelques travaux recents ont tente, grace a l'utilisation des isotopes radioactifs, de dresser le bilan des flux d'entree et de sortie des ions a travers les membranes permeables in vivo, alors que les methodes d'analyse chimique permettent seulement de dresser le bilan net de ces

  12. An Atlas for Schistosoma mansoni Organs and Life-Cycle Stages Using Cell Type-Specific Markers and Confocal Microscopy

    Science.gov (United States)

    Cogswell, Alexis; Williams, David L.; Newmark, Phillip A.

    2011-01-01

    Schistosomiasis (bilharzia) is a tropical disease caused by trematode parasites (Schistosoma) that affects hundreds of millions of people in the developing world. Currently only a single drug (praziquantel) is available to treat this disease, highlighting the importance of developing new techniques to study Schistosoma. While molecular advances, including RNA interference and the availability of complete genome sequences for two Schistosoma species, will help to revolutionize studies of these animals, an array of tools for visualizing the consequences of experimental perturbations on tissue integrity and development needs to be made widely available. To this end, we screened a battery of commercially available stains, antibodies and fluorescently labeled lectins, many of which have not been described previously for analyzing schistosomes, for their ability to label various cell and tissue types in the cercarial stage of S. mansoni. This analysis uncovered more than 20 new markers that label most cercarial tissues, including the tegument, the musculature, the protonephridia, the secretory system and the nervous system. Using these markers we present a high-resolution visual depiction of cercarial anatomy. Examining the effectiveness of a subset of these markers in S. mansoni adults and miracidia, we demonstrate the value of these tools for labeling tissues in a variety of life-cycle stages. The methodologies described here will facilitate functional analyses aimed at understanding fundamental biological processes in these parasites. PMID:21408085

  13. An artemisinin derivative of praziquantel as an orally active antischistosomal agent.

    Directory of Open Access Journals (Sweden)

    Lanlan Dong

    Full Text Available Schistosomiasis is a major health problem in tropical and sub-tropical areas caused by species of trematode belonging to the genus Schistosoma. The treatment and control of this disease has been relying on the use of a single drug praziquantel. However, the drug resistance concern urged the development of new drugs against schistosoma. Here, we report our systematic biological evaluation of DW-3-15, a new lead compound developed based on our conjugation design rationale as an effective anti-schistosomal agent.The antischistosomal activity of DW-3-15 was systematically evaluated in S. japonicum infected mouse model for its stage-sensitivity and dose response. The results revealed that DW-3-15 exhibited 60-85% worm reduction rate against different development stage of worm. Scanning electron microscopy (SEM observation indicated that DW-3-15 may damage to the tegument of male schistosomes.Our results demonstrated that DW-3-15 showed potent anti-schistosomal activities in vivo. The results strongly support our conjugation design strategy of artemisinin analogs and further development of DW-3-15 as a new lead compound as anti-schistosomal agent.

  14. Identification of lead chemotherapeutic agents from medicinal plants against blood flukes and whipworms.

    Science.gov (United States)

    Wangchuk, Phurpa; Giacomin, Paul R; Pearson, Mark S; Smout, Michael J; Loukas, Alex

    2016-08-30

    Schistosomiasis and trichuriasis are two of the most common neglected tropical diseases (NTD) that affect almost a billion people worldwide. There is only a limited number of effective drugs to combat these NTD. Medicinal plants are a viable source of parasiticides. In this study, we have investigated six of the 19 phytochemicals isolated from two Bhutanese medicinal plants, Corydalis crispa and Pleurospermum amabile, for their anthelmintic properties. We used the xWORM technique and Scanning Electron Microscope-based imaging to determine the activity of the compounds. Of the six compounds tested, isomyristicin and bergapten showed significant anthelmintic activity against Schistosoma mansoni and Trichuris muris with bergapten being the most efficacious compound one against both parasites (S. mansoni IC50 = 8.6 μg/mL and T. muris IC50 = 10.6 μg/mL) and also against the schistosomulum stage of S. mansoni. These two compounds induced tegumental damage to S. mansoni and affected the cuticle, bacillary bands and bacillary glands of T. muris. The efficacy against multiple phylogenetically distinct parasites and different life stages, especially the schistosomulum where praziquantel is ineffective, makes isomyristicin and bergapten novel scaffolds for broad-spectrum anthelmintic drug development that could be used for the control of helminths infecting humans and animals.

  15. Effects of dietary intake of garlic on intestinal trematodes.

    Science.gov (United States)

    Cortés, Alba; García-Ferrús, Miguel; Sotillo, Javier; Guillermo Esteban, J; Toledo, Rafael; Muñoz-Antolí, Carla

    2017-08-01

    The current strategy for the control of helminth infections relies on chemotherapy. However, resistance appearance is promoting the necessity of developing new drugs against trematodes. Herein, potential trematocidal effects of garlic (Allium sativum) are investigated in the context of intestinal foodborne trematodes, employing the Echinostoma caproni-mouse model. Daily administration of dietary doses of garlic was conducted in three groups of mice: (i) before infection (prophylaxis), (ii) after infection (therapeutic) and (iii) both, before and after infection (continuous). A fourth group of mice, not exposed to garlic, was used as control. No differences in worm recovery, fecundity and local cytokine expression profiles were found with respect to control infections. However, considerable alterations in tegument structure, including swelling, furrowing, vacuolization and changes in secretory bodies were detected in garlic-exposed parasites using scanning and transmission electron microscopy. Protein secretion was markedly reduced in response to garlic, whereas up-regulation of several proteins, such as major vault protein and tER-ATPase, was observed in treated worms. The results presented herein provide new insights in the anthelminthic activity of bioactive garlic compounds and the manner that parasites respond to toxins.

  16. [Responses of functional diversity of aquatic insect community to land use change in middle reach of Qiantang River, East China].

    Science.gov (United States)

    Zhang, Lian-Bo; Liu, Dong-Xiao; Liu, Shuo-Ru; Zhang, Yong; Tong, Xiao-Li; Wang, Bei-Xin

    2013-10-01

    Based on the biological traits such as life history, resistance ability against environmental disturbance, and physiological characteristics of aquatic insects, and by using the fourth-corner statistical method, this paper studied the responses of the functional diversity of aquatic insect community to land use change in the middle reach of Qiantang River, Zhejiang Province of East China. For the test aquatic insect community, some of its biological traits were sensitive to land use change, and altered along human disturbance gradients as expected. With the increasing intensity of human disturbance, the maximal insect body length decreased gradually, the dominant respiration pattern evolved from gill respiration to tegument respiration, and the abundance of burrowers increased significantly. At the same time, the functional diversity measured as Rao's quadratic entropy was significantly higher in reference sites than in disturbed sites (P aquatic community were mainly induced by the land use change caused by human activities, which resulted in the decline of stream water quality and habitat quality and the variations of aquatic insect community composition and biological traits. The aquatic insect biological traits and functional diversity could be the potentially effective indicators in the stream health assessment in the future.

  17. Visualization of the dynamic multimerization of human Cytomegalovirus pp65 in punctuate nuclear foci

    International Nuclear Information System (INIS)

    Cui Zongqiang; Zhang Ke; Zhang Zhiping; Liu Yalan; Zhou Yafeng; Wei Hongping; Zhang Xian-En

    2009-01-01

    The phosphorylated protein pp65 of human Cytomegalovirus (HCMV) is the predominant virion protein and the major tegument constituent. It plays important roles in HCMV infection and virion assembly. Live cell imaging and fluorescence recovery after photobleaching (FRAP) analysis showed that HCMV pp65 accumulated dynamically in punctuate nuclear foci when transiently expressed in mammalian cells. Fluorescence resonance energy transfer (FRET) imaging disclosed that pp65 can self-interact in its localization foci. Yeast two-hybrid assay verified that pp65 is a self-associating protein, and the N-terminal amino acids 14-22 were determined to be essential for pp65 self-association. However, these amino acids were not related to pp65 localization in the specific nuclear foci. The interaction of pp65 and ppUL97 was also studied by FRET microscopy, and the result suggested that there is another signal sequence in pp65, being the ppUL97 phosphorylation site, that is responsible for localization of pp65 in nuclear foci. These results help to understand the function of pp65 in HCMV infection and virion morphogenesis.

  18. Immunological and biochemical studies of fascioliasis in goats and cattle

    Energy Technology Data Exchange (ETDEWEB)

    Reddington, J.J.

    1985-01-01

    Using the goat as a susceptible host and cattle as a resistant species to Fasciola hepatica infections, the humoral response of these animals to the surface of the newly excysted juvenile (NEJ) fluke was examined. Tegumental proteins of the NEJ were labeled with /sup 125/I by lactoperoxidase and analyzed after immunoprecipitation using a double antibody system. In addition, a comparison was made between the infected sera's capacity to immunoprecipitate surface antigens and their in vitro cytotoxic activity against the NEJ. In both goats and cattle the levels of NEJ surface antigens precipitated increased during the first 4 weeks PI. The peak immunoprecipitation of NEJ surface antigens by cattle sera (58%) was significantly higher than that of infected goat sera (33%). Immunoprecipitation of the available radiolabeled NEJ surface proteins by the infected cattle sera remained consistently higher than goat sera until the 16th week PI. The cytotoxic effects of these same caprine sera on NEJs in vitro was limited, while the cytotoxicity of the infected bovine sera closely approximated the sera's ability to precipitate NEJ surface antigens. There was also a qualitative difference between the species in their recognition of /sup 35/S and /sup 125/I radiolabeled NEJ surface antigens. Uninfected goat or cattle sera failed to precipitate any /sup 125/I or /sup 35/S-labeled surface proteins.

  19. Localization of serotonin and ultrastructure of serotonergic neutrons in the nervous system of fasciola hepatica

    International Nuclear Information System (INIS)

    Huang Shile; Cheng Bing; Rong Yaofang

    1993-01-01

    Rabbits antisera were raised against an antigen prepared by coupling 5-HT to bovine serum albumin (BSA) using formaldehyde as a coupling reagent. The fresh adult Fasciola hepatica were fixed with 4% formaldehyde and sectioned on a cryostat. The sections were stained by indirect immunofluorescence technique. Abundant immunofluorescence specific for 5-HT was observed in ganglion cell bodies and their processes, the transverse commissure that connects two ganglia and longitudinal axes extending from the ganglia. Immuno-reactivity to 5-HT was also found in the nerve fibre innervating tegument, gut wall, the epithelium of testes or ovary, the musculature of uterus and ootype, etc. The ultrastructure of serotonergic neurons was visualized. As in other invertebrates, the serotonergic neutrons of Fasciola hepatica consisted of cell bodies, axons, synapses, herring bodies and neuromuscular junctions. The nerve cell bodies were aggregatively located in ganglia and many dispersed spherical granular vesicles were present in cytoplasm. The nerve axons branched out to the muscles forming synapses, where synaptic vesicles contained 5-HT dense-core granules were found. The distribution of 5-HT within the neurons strongly suggested that 5-HT was functioning as a neurotrasmitter in Fasciola hepatica

  20. [Preclinical studies of cucurbita maxima (pumpkin seeds) a traditional intestinal antiparasitic in rural urban areas].

    Science.gov (United States)

    Díaz Obregón, Daysi; Lloja Lozano, Luis; Carbajal Zúñiga, Victor

    2004-01-01

    Experimental research was carried out at the Parasitology and Chemistry laboratories of the Jorge Basadre Grohmann National University, in Tacna. The process involved two phases: (1) determination of the minimum inhibitory concentration (MIC) of Cucurbita Maxima as an antiparasitic agent using canine tapeworms with an intestinal isolation of 5 to 6 hours, and (2) determination of the side-effects of Curbita Maxima on exposed albino rats. It was found that the MIC of 23 gr. of pumpkin seed in 100 ml. of distilled water can produce an antihelminthic effect. This concentration is equivalent to +/- 73 pumpkin seeds (x2 = 5.6, p 23 gr. There is a protheolithic effect with an average survival time of 38.4 minutes. Microscopically the mature proglottids present a destruction of the tegument involving the basal membrane. In the gravid proglottids there is egg destruction. These findings are accentuated when experimenting with Cucurbita Maxima in a concentration of 30 and 32 gr. Superficial non-erosive gastritis was found in weys rats after 4 hours of administering 9 gr/kg.

  1. The life cycle of Ascocotyle (Phagicola) longa (Digenea: Heterophyidae), a causative agent of fish-borne trematodosis.

    Science.gov (United States)

    Simões, Susana Balmant Emerique; Barbosa, Helene Santos; Santos, Cláudia Portes

    2010-03-01

    The complete life cycle of the trematode Ascocotyle (Phagicola) longa (Digenea: Heterophyidae) is elucidated by natural observation validated by experimental infections. The natural first intermediate host of A. (P.) longa, an agent of human heterophyiasis in Brazil, is the cochliopid snail Heleobia australis (new first intermediate host). Metacercariae were found encysted in the body musculature, heart, stomach, liver, kidney, spleen, gonads and mesentery of mullets Mugil liza. Hamsters Mesocricetus auratus were experimentally infected with metacercariae of A. (P.) longa obtained from the mullets, and the adults recovered were used to infect the snails H. australis. Rediae and cercariae of A. (P.) longa are described for the first time. The ultrastructure of the tegument of A. (P.) longa shows a change in spination pattern from the cercaria with single-pointed spines to the metacercaria and adult with multipointed, brush-shaped spines. The life cycle of A. (P.) longa is related to estuaries and coastal lagoons where the recruitment of mugilid juveniles occurs. The high prevalence (100%) of A. (P.) longa encysted in the mullets examined within the urban area of Rio de Janeiro indicates the potentially great public health impact of the consumption of raw mullets. Copyright 2009 Elsevier B.V. All rights reserved.

  2. Curcumin Generates Oxidative Stress and Induces Apoptosis in Adult Schistosoma mansoni Worms.

    Directory of Open Access Journals (Sweden)

    Daniela de Paula Aguiar

    Full Text Available Inducing apoptosis is an interesting therapeutic approach to develop drugs that act against helminthic parasites. Researchers have investigated how curcumin (CUR, a biologically active compound extracted from rhizomes of Curcuma longa, affects Schistosoma mansoni and several cancer cell lines. This study evaluates how CUR influences the induction of apoptosis and oxidative stress in couples of adult S. mansoni worms. CUR decreased the viability of adult worms and killed them. The tegument of the parasite suffered morphological changes, the mitochondria underwent alterations, and chromatin condensed. Different apoptotic parameters were determined in an attempt to understand how CUR affected adult S. mansoni worms. CUR induced DNA damage and fragmentation and increased the expression of SmCASP3/7 transcripts and the activity of Caspase 3 in female and male worms. However, CUR did not intensify the activity of Caspase 8 in female or male worms. Evaluation of the superoxide anion and different antioxidant enzymes helped to explore the mechanism of parasite death further. The level of superoxide anion and the activity of Superoxide Dismutase (SOD increased, whereas the activity of Glutathione-S-Transferase (GST, Glutathione reductase (GR, and Glutathione peroxidase (GPX decreased, which culminated in the oxidation of proteins in adult female and male worms incubated with CUR. In conclusion, CUR generated oxidative stress followed by apoptotic-like-events in both adult female and male S. mansoni worms, ultimately killing them.

  3. Morfologia externa de espécimes adultos de Paratanaisia bragai (Santos, 1934 (Digenea: Eucotylidae External morphology of the adults specimens of Paratanaisia bragai (Santos, 1934(Diginea; Eucotylidae

    Directory of Open Access Journals (Sweden)

    Solange V.P.B. Brandolini

    2007-09-01

    Full Text Available Espécimes adultos de Paratanaisia bragai foram coletados de ductos renais de Columba livia infectados naturalmente, fixados em glutaraldeído 2,5% e processados para microscopia eletrônica de varredura. Foram identificados dois tipos distintos de escamas, escamas bífidas e simples, dispostas em fileiras transversais no tegumento das faces dorsal e ventral do corpo. Também mostrando em detalhes a ventosa oral subterminal, bem desenvolvida e acetábulo de tamanho reduzido, localizado no terço médio do corpo, circundado por pequenas protuberâncias e quatro papilas dispostas regularmente na sua margem.Adult specimens of Paratanaisia bragai collected from renal ducts of Columba livia naturally infected, were fixed in glutaraldehyde 2.5% and processed for scanning electron microscopy. Two distinct types of scales had been identified, bifid and simple scales, disposed in transversal rows, on the tegument of dorsal and ventral face of the body. Also show in details the oral sucker subterminal, well developed and acetabulum of size reduced, located in third medium of the body, surrounded for small protuberance and four papillae on its edge.

  4. The Us2 gene product of herpes simplex virus 2 is a membrane-associated ubiquitin-interacting protein.

    Science.gov (United States)

    Kang, Ming-Hsi; Roy, Bibhuti B; Finnen, Renée L; Le Sage, Valerie; Johnston, Susan M; Zhang, Hui; Banfield, Bruce W

    2013-09-01

    The Us2 gene encodes a tegument protein that is conserved in most members of the Alphaherpesvirinae. Previous studies on the pseudorabies virus (PRV) Us2 ortholog indicated that it is prenylated, associates with membranes, and spatially regulates the enzymatic activity of the MAP (mitogen-activated protein) kinase ERK (extracellular signal-related kinase) through direct binding and sequestration of ERK at the cytoplasmic face of the plasma membrane. Here we present an analysis of the herpes simplex virus 2 (HSV-2) Us2 ortholog and demonstrate that, like PRV Us2, HSV-2 Us2 is a virion component and that, unlike PRV Us2, it does not interact with ERK in yeast two-hybrid assays. HSV-2 Us2 lacks prenylation signals and other canonical membrane-targeting motifs yet is tightly associated with detergent-insoluble membranes and localizes predominantly to recycling endosomes. Experiments to identify cellular proteins that facilitate HSV-2 Us2 membrane association were inconclusive; however, these studies led to the identification of HSV-2 Us2 as a ubiquitin-interacting protein, providing new insight into the functions of HSV-2 Us2.

  5. Identification of pathogens for differential diagnosis of fever with jaundice in the Central African Republic: a retrospective assessment, 2008-2010.

    Science.gov (United States)

    Gadia, Christelle Luce Bobossi; Manirakiza, Alexandre; Tekpa, Gaspard; Konamna, Xavier; Vickos, Ulrich; Nakoune, Emmanuel

    2017-11-29

    Febrile jaundice results clinically in generalized yellow coloration of the teguments and mucous membranes due to excess plasma bilirubin, accompanied by fever. Two types are found: conjugated and unconjugated bilirubin jaundice. Jaundice is a sign in several diseases due to viruses (viral hepatitis and arbovirus), parasites (malaria) and bacteria (leptospirosis). In the Central African Republic (CAR), only yellow fever is included on the list of diseases for surveillance. The aim of this study was to identify the other pathogens that can cause febrile jaundice, for better management of patients. Between 2008 and 2010, 198 sera negative for yellow fever IgM were randomly selected from 2177 samples collected during yellow fever surveillance. Laboratory analyses targeted four groups of pathogens: hepatitis B, C, delta and E viruses; dengue, chikungunya, Zika, Crimean-Congo haemorrhagic fever, West Nile and Rift Valley arboviruses; malaria parasites; and bacteria (leptospirosis). Overall, 30.9% sera were positive for hepatitis B, 20.2% for hepatitis E, 12.3% for hepatitis C and 8.2% for malaria. The majority of positive sera (40.4%) were from people aged 16-30 years. Co-infection with at least two of these pathogens was also found. These findings suggest that a systematic investigation should be undertaken of infectious agents that cause febrile jaundice in the CAR.

  6. Les acides gras hydroxylés : agro-tensioactifs aux propriétés moussantes originales

    Directory of Open Access Journals (Sweden)

    Fameau Anne-Laure

    2013-01-01

    Full Text Available Surfactants, which are molecules bearing a hydrophobic part and a hydrophilic part, are widely used in many daily life products (detergent, cosmetic, pharmaceutical... because of their foaming properties. Indeed, they spontaneously go to the interface between air and water, which stabilizes the thin water films surrounding air bubbles to obtain foams. Currently, since the most used surfactants come from derivatives from petroleum-based products, their replacement by biomolecules is a major issue in the current context of sustainable development and green chemistry. This could promote the use of molecules from agricultural resources, such as hydroxyl fatty acids that can be extracted from oil or from the tegument of fruits and vegetables, as new agro-based surfactants. Here, we present recent research on the surfactant properties of the 12-hydroxy stearic acid coming from the castor oil. This fatty acid has very advantageous foaming properties compared to conventional synthetic surfactant systems: it enables the production of foams which are at the same time very abundant and, remarkably, stables over a span lasting more than 6 months. Moreover, these foams are thermoresponsive because the foam stability can be easily modulated by a simple modification of the temperature.

  7. Directory of Open Access Journals (Sweden)

    I. Gabanyi

    2013-01-01

    Full Text Available The type I herpes simplex virus VP22 tegument protein is abundant and well known for its ability to translocate proteins from one cell to the other. In spite of some reports questioning its ability to translocate proteins by attributing the results observed to fixation artifacts or simple attachment to the cell membrane, VP22 has been used to deliver several proteins into different cell types, triggering the expected cell response. However, the question of the ability of VP22 to enter stem cells has not been addressed. We investigated whether VP22 could be used as a tool to be applied in stem cell research and differentiation due to its capacity to internalize other proteins without altering the cell genome. We generated a VP22.eGFP construct to evaluate whether VP22 could be internalized and carry another protein with it into two different types of stem cells, namely adult human dental pulp stem cells and mouse embryonic stem cells. We generated a VP22.eGFP fusion protein and demonstrated that, in fact, it enters stem cells. Therefore, this system may be used as a tool to deliver various proteins into stem cells, allowing stem cell research, differentiation and the generation of induced pluripotent stem cells in the absence of genome alterations.

  8. The virion-associated open reading frame 49 of murine gammaherpesvirus 68 promotes viral replication both in vitro and in vivo as a derepressor of RTA.

    Science.gov (United States)

    Noh, Cheol-Woo; Cho, Hye-Jeong; Kang, Hye-Ri; Jin, Hyun Yong; Lee, Shaoying; Deng, Hongyu; Wu, Ting-Ting; Arumugaswami, Vaithilingaraja; Sun, Ren; Song, Moon Jung

    2012-01-01

    Replication and transcription activator (RTA), an immediate-early gene, is a key molecular switch to evoke lytic replication of gammaherpesviruses. Open reading frame 49 (ORF49) is conserved among gammaherpesviruses and shown to cooperate with RTA in regulating virus lytic replication. Here we show a molecular mechanism and in vivo functions of murine gammaherpesvirus 68 (MHV-68 or γHV-68) ORF49. MHV-68 ORF49 was transcribed and translated as a late gene. The ORF49 protein was associated with a virion, interacting with the ORF64 large tegument protein and the ORF25 capsid protein. Moreover, ORF49 directly bound to RTA and its negative cellular regulator, poly(ADP-ribose) polymerase-1 (PARP-1), and disrupted the interactions of RTA and PARP-1. Productive replication of an ORF49-deficient mutant virus (49S) was attenuated in vivo as well as in vitro. Likewise, latent infection was also impaired in the spleen of 49S-infected mice. Taken together, our results suggest that the virion-associated ORF49 protein may promote virus replication both in vitro and in vivo by providing an optimal environment in the early phase of virus infection as a derepressor of RTA.

  9. Antischistosomal activity of a calcium channel antagonist on schistosomula and adult Schistosoma mansoni worms

    Directory of Open Access Journals (Sweden)

    Vanessa Silva-Moraes

    2013-08-01

    Full Text Available Current schistosomiasis control strategies are largely based on chemotherapeutic agents and a limited number of drugs are available today. Praziquantel (PZQ is the only drug currently used in schistosomiasis control programs. Unfortunately, this drug shows poor efficacy in patients during the earliest infection phases. The effects of PZQ appear to operate on the voltage-operated Ca2+channels, which are located on the external Schistosoma mansoni membrane. Because some Ca2+channels have dihydropyridine drug class (a class that includes nifedipine sensitivity, an in vitro analysis using a calcium channel antagonist (clinically used for cardiovascular hypertension was performed to determine the antischistosomal effects of nifedipine on schistosomula and adult worm cultures. Nifedipine demonstrated antischistosomal activity against schistosomula and significantly reduced viability at all of the concentrations used alone or in combination with PZQ. In contrast, PZQ did not show significant efficacy when used alone. Adult worms were also affected by nifedipine after a 24 h incubation and exhibited impaired motility, several lesions on the tegument and intense contractility. These data support the idea of Ca2+channels subunits as drug targets and favour alternative therapeutic schemes when drug resistance has been reported. In this paper, strong arguments encouraging drug research are presented, with a focus on exploring schistosomal Ca2+channels.

  10. Synthesis and anthelmintic activity of arctigenin derivatives against Dactylogyrus intermedius in goldfish.

    Science.gov (United States)

    Hu, Yang; Liu, Lei; Liu, Guang-Lu; Tu, Xiao; Wang, Gao-Xue; Ling, Fei

    2017-08-01

    To control the parasitic disease of Dactylogyrus intermedius, a series of new arctigenin derivatives were designed, synthesized and tested in our study. The anthelmintic activity of most of the derivatives ranged from 1 to 10mg/L. Compared to traditional drug praziquantel (EC 50 =2.69mg/L), ether derivatives 2g and 2h exhibited slightly higher anti-parasitic activity, with the EC 50 values of 2.48 and 1.52mg/L, respectively. Furthermore, the arctigenin-imidazole hybrids 4a and 4b also removed D. intermedius effectively, with the EC 50 values of 2.13 and 2.07mg/L, respectively. The structure-activity relationship analysis indicated that four carbon atoms length of linker and imidazole substitute group could significantly increase the anthelmintic activity, and reduced the toxicity. Through the scanning electron microscope observation, compounds 4a and 4b caused the D. intermedius tegumental damage such as intensive wrinkles, holes and nodular structures. Overall, the structural optimization analysis of arctigenin suggested that 4a and 4b can be used for preventing and controlling Dactylogyrus infections and considered as promising lead compounds for the development of commercial drugs. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. VP22 herpes simplex virus protein can transduce proteins into stem cells

    Energy Technology Data Exchange (ETDEWEB)

    Gabanyi, I.; Lojudice, F.H.; Kossugue, P.M. [Centro de Terapia Celular e Molecular, Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, SP (Brazil); Rebelato, E. [Departamento de Fisiologia e Biofísica, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP (Brazil); Demasi, M.A.; Sogayar, M.C. [Centro de Terapia Celular e Molecular, Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, SP (Brazil)

    2013-02-01

    The type I herpes simplex virus VP22 tegument protein is abundant and well known for its ability to translocate proteins from one cell to the other. In spite of some reports questioning its ability to translocate proteins by attributing the results observed to fixation artifacts or simple attachment to the cell membrane, VP22 has been used to deliver several proteins into different cell types, triggering the expected cell response. However, the question of the ability of VP22 to enter stem cells has not been addressed. We investigated whether VP22 could be used as a tool to be applied in stem cell research and differentiation due to its capacity to internalize other proteins without altering the cell genome. We generated a VP22.eGFP construct to evaluate whether VP22 could be internalized and carry another protein with it into two different types of stem cells, namely adult human dental pulp stem cells and mouse embryonic stem cells. We generated a VP22.eGFP fusion protein and demonstrated that, in fact, it enters stem cells. Therefore, this system may be used as a tool to deliver various proteins into stem cells, allowing stem cell research, differentiation and the generation of induced pluripotent stem cells in the absence of genome alterations.

  12. First ultrastructural data on the human tapeworm Taenia asiatica eggs by scanning and transmission electron microscopy (SEM, TEM).

    Science.gov (United States)

    Galán-Puchades, M Teresa; Yang, Yichao; Marcilla, Antonio; Choe, Seongjun; Park, Hansol; Osuna, Antonio; Eom, Keeseon S

    2016-09-01

    Humans are definitive hosts of three species of the Taenia genus, namely Taenia solium, Taenia saginata and Taenia asiatica. The relative novelty of the latter explains the lack of knowledge concerning certain relevant aspects related to this parasite, such as its definite geographical distribution and whether its eggs can infect humans or not. So far, only the eggs of T. solium are known to be infective for humans, producing cysticercosis. Although eggs contain the infective stage, the oncosphere, there is a lack of research on the ultrastructure of eggs of human taeniids. We show, for the first time, the ultrastructure of eggs of T. asiatica by means of SEM and TEM analyses. We detected all the envelopes, namely the egg shell, vitelline layer, outer embryophoric membrane, embryophore, granular layer, basal membrane, oncospheral membrane and oncospheral tegument. Hooks surrounded by myofibrils and glycogen-like particles, the two types of secretory granules of the penetration glands, as well as several nuclei and mitochondria were also revealed in the oncospheres. In addition to the already known structures in eggs from other Taenia species, the presence of two types of small vesicles is described herein, possibly corresponding to exosomes and ectosomes because of their shape and size, which could participate in the host/parasite intercellular communication.

  13. Cloning and characterization of the fatty acid-binding protein gene from the protoscolex of Taenia multiceps.

    Science.gov (United States)

    Nie, Hua-Ming; Xie, Yue; Fu, Yan; Yang, Ying-Dong; Gu, Xiao-Bin; Wang, Shu-Xian; Peng, Xi; Lai, Wei-Ming; Peng, Xue-Rong; Yang, Guang-You

    2013-05-01

    Taenia multiceps (Cestoda: Taeniidae), a worldwide cestode parasite, is emerging as an important helminthic zoonosis due to serious or fatal central nervous system disease commonly known as coenurosis in domestic and wild ruminants including humans. Herein, a fatty acid-binding protein (FABP) gene was identified from transcriptomic data in T. multiceps. This gene, which contains a complete coding sequence, was amplified by reverse-transcriptase polymerase chain reaction. The corresponding protein, which was named TmFABP, had a molecular weight of 14 kDa, and subsequently was recombinantly expressed in Escherichia coli. The fusion protein was purified on Ni-NTA beads (Bio-Rad). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analyses showed that the purified recombinant protein caused immunogenicity. Immunohistochemical studies showed that TmFABP was expressed at the tegumental level in the protoscolices and in the cells between the body wall and parenchyma layer of the cestode. In sections from gravid proglottids, intense staining was detected in the uterus and eggs. Based on this, TmFABP could be switched on during differentiation of germinative layers to protoscoleces and from metacestodes to adult worms. Taken together, our results already reported for T. multiceps suggest the possibility of TmFABP developing a vaccine to control and prevent coenurosis.

  14. Histological and ultrastructural localization of antigen B in the metacestode of Taenia solium

    Energy Technology Data Exchange (ETDEWEB)

    Laclette, J.P.; Merchant, M.T.; Willms, K.

    1987-02-01

    The morphological localization of antigen B (AgB) in the tissues of the Taenia solium metacestode was studied by immunological and biochemical methods. Indirect immunofluorescence carried out on vibratome sections showed that AgB is widely distributed throughout the tissue. A more intense fluorescence was observed in the tegumentary cytons of the bladder wall and in the lumen of the spiral canal of the invaginated scolex. Ultrastructural analysis of larvae washed in PBS after dissection from meat and then incubated with rabbit antibodies against AgB, followed by peroxidase-labeled goat anti-rabbit IgG, did not exhibit electron-dense material on the external surface. Larvae fixed in glutaraldehyde immediately after dissection and exposed to the immunoperoxidase reagents did exhibit electron-dense material on microtriches, indicating that AgB is only loosely bound to the external surface. Crude extracts of surface-radioiodinated cysticerci analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) contained no labeled proteins with the molecular weight of AgB. Autoradiography of the immunoelectrophoretograms in which the crude extract was confronted with antibodies to AgB demonstrated that this antigen was not labeled, and therefore is not exposed on the tegumentary surface. The results suggest that AgB is synthesized by the tegumentary cytons of the parasite and secreted through the tegumental membrane into the host tissues and the lumen of the spiral canal.

  15. In vitro ovicidal and cestocidal effects of toxins from Bacillus thuringiensis on the canine and human parasite Dipylidium caninum.

    Science.gov (United States)

    Peña, Guadalupe; Aguilar Jiménez, Fortino Agustín; Hallal-Calleros, Claudia; Morales-Montor, Jorge; Hernández-Velázquez, Víctor Manuel; Flores-Pérez, Fernando Iván

    2013-01-01

    Bacillus thuringiensis is a gram-positive soil-dwelling bacterium that is commonly used as a biological pesticide. This bacterium may also be used for biological control of helminth parasites in domestic animals. In this study, we evaluated the possible ovicidal and cestocidal effects of a total protein extract of B. thuringiensis native strains on the zoonotic cestode parasite of dogs, Dipylidium caninum (D. caninum). Dose and time response curves were determined by coincubating B. thuringiensis proteins at concentration ranging from 100 to 1000 μ g/mL along with 4000 egg capsules of D. caninum. Egg viability was evaluated using the trypan blue exclusion test. The lethal concentration of toxins on eggs was 600 μ g/ml, and the best incubation time to produce this effect was 3 h. In the adult stage, the motility and the thickness of the tegument were used as indicators of damage. The motility was inhibited by 100% after 8 hours of culture compared to the control group, while the thickness of the cestode was reduced by 34%. Conclusively, proteins of the strain GP526 of B. thuringiensis directly act upon D. caninum showing ovicidal and cestocidal effects. Thus, B. thuringiensis is proposed as a potential biological control agent against this zoonosis.

  16. In Vitro Ovicidal and Cestocidal Effects of Toxins from Bacillus thuringiensis on the Canine and Human Parasite Dipylidium caninum

    Directory of Open Access Journals (Sweden)

    Guadalupe Peña

    2013-01-01

    Full Text Available Bacillus thuringiensis is a gram-positive soil-dwelling bacterium that is commonly used as a biological pesticide. This bacterium may also be used for biological control of helminth parasites in domestic animals. In this study, we evaluated the possible ovicidal and cestocidal effects of a total protein extract of B. thuringiensis native strains on the zoonotic cestode parasite of dogs, Dipylidium caninum (D. caninum. Dose and time response curves were determined by coincubating B. thuringiensis proteins at concentration ranging from 100 to 1000 μg/mL along with 4000 egg capsules of D. caninum. Egg viability was evaluated using the trypan blue exclusion test. The lethal concentration of toxins on eggs was 600 μg/ml, and the best incubation time to produce this effect was 3 h. In the adult stage, the motility and the thickness of the tegument were used as indicators of damage. The motility was inhibited by 100% after 8 hours of culture compared to the control group, while the thickness of the cestode was reduced by 34%. Conclusively, proteins of the strain GP526 of B. thuringiensis directly act upon D. caninum showing ovicidal and cestocidal effects. Thus, B. thuringiensis is proposed as a potential biological control agent against this zoonosis.

  17. Acute schistosomiasis diagnosis: a new tool for the diagnosis of schistosomiasis in a group of travelers recently infected in a new focus of Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    Rafaella Fortini Queiroz Grenfell

    2013-04-01

    Full Text Available Introduction The diagnosis of schistosomiasis mansoni on early stages of infection is important to prevent late morbidity. A simple, cheap, sensitive and specific assay for routine diagnosis of schistosome infection based on the detection of specific IgG for schistosomula tegument antigens (ELISA-SmTeg was developed by our group. Methods We describe here an acute outbreak involving a travel group of 80 individuals from a non-endemic area of the State of Minas Gerais, Brazil. These individuals were in contact with a freshwater pool where Biomphalaria glabrata was found. Results obtained from our new methodology were compared to IgG antibody titers against soluble worm antigenic preparation (SWAP by ELISA and, also to parasitological examination, nuclear magnetic resonance and clinical findings. Results ELISA-SmTeg was capable of detecting 64 positive cases among the 80 individuals participating at the survey with a positivity ratio of 80% and a higher sensitivity than ELISA-SWAP that was only sensitive for 56% of positive cases. Besides, a significant correlation was found for the severity of the infection and the specific IgG titers against SmTeg. Conclusions Our data showed that ELISA-SmTeg might serve as the initial diagnostic tool for acute stages of the infection in community-based helminth control programs or for the surveillance of individuals from non-endemic areas.

  18. Monoclonal antibody-based dipstick assay: a reliable field applicable technique for diagnosis of Schistosoma mansoni infection using human serum and urine samples.

    Science.gov (United States)

    Demerdash, Zeinab; Mohamed, Salwa; Hendawy, Mohamed; Rabia, Ibrahim; Attia, Mohy; Shaker, Zeinab; Diab, Tarek M

    2013-02-01

    A field applicable diagnostic technique, the dipstick assay, was evaluated for its sensitivity and specificity in diagnosing human Schistosoma mansoni infection. A monoclonal antibody (mAb) against S. mansoni adult worm tegumental antigen (AWTA) was employed in dipstick and sandwich ELISA for detection of circulating schistosome antigen (CSA) in both serum and urine samples. Based on clinical and parasitological examinations, 60 S. mansoni-infected patients, 30 patients infected with parasites other than schistosomiasis, and 30 uninfected healthy individuals were selected. The sensitivity and specificity of dipstick assay in urine samples were 86.7% and 90.0%, respectively, compared to 90.0% sensitivity and 91.7% specificity of sandwich ELISA. In serum samples, the sensitivity and specificity were 88.3% and 91.7% for dipstick assay vs. 91.7% and 95.0% for sandwich ELISA, respectively. The diagnostic efficacy of dipstick assay in urine and serum samples was 88.3% and 90.0%, while it was 90.8% and 93.3% for sandwich ELISA, respectively. The diagnostic indices of dipstick assay and ELISA either in serum or in urine were statistically comparable (P>0.05). In conclusion, the dipstick assay offers an alternative simple, rapid, non-invasive technique in detecting CSA or complement to stool examinations especially in field studies.

  19. Ultrastructural and biochemical detection of biotin and biotinylated polypeptides in Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    Santos P.R.P.

    1997-01-01

    Full Text Available Biotinylation is proposed for the identification of surface proteins in Schistosoma mansoni using the streptavidin-HRP conjugate for the detection of labeled polypeptides. However, control samples also showed several endogenous biotinylated polypeptides. In an attempt to determine the possibility of nonspecific binding between the streptavidin-HRP conjugate and polypeptides from S. mansoni, the conjugate was blocked with biotinamidecaproate-N-hydroxysuccinimide ester (BcapNHS before biotin-streptavidin blotting. No bands were detected on the nitrocellulose sheet, demonstrating the specific recognition of biotin by the streptavidin present in the conjugate. Whole cercariae and cercarial bodies and tails showed several endogenous biotinylated polypeptides. The biotin concentration was 13 µg/190,000 cercariae. Adult worms presented less endogenous biotinylated polypeptides than cercariae. These results may be due to changes in the environment from aerobic to anaerobic conditions when cercarial bodies (schistosomula are transformed into adult worms and a decrease in CO2 production may occur. Cercariae, cercarial bodies and adult male worms were examined by transmission electron microscopy employing an avidin-colloidal gold conjugate for the detection of endogenous biotin. Gold particles were distributed mainly on the muscle fibers, but dispersed granules were observed in the tegument, mitochondria and cytosol. The discovery of endogenous biotin in S. mansoni should be investigated in order to clarify the function of this vitamin in the parasite

  20. Anticestodal Activity of Endophytic Pestalotiopsis sp. on Protoscoleces of Hydatid Cyst Echinococcus granulosus

    Directory of Open Access Journals (Sweden)

    Vijay C. Verma

    2013-01-01

    Full Text Available Surgery is still the main treatment in hydatidosis caused by Echinococcus, which is a global health problem in human and animals. So, there is need for some natural protoscolicidal agents for instillation to prevent their reoccurrence at therapeutic doses. In this present investigation, anticestodal activity of one of the endophytic fungi Pestalotiopsis sp. from Neem plant was observed on protoscoleces of hydatid cysts of Echinococcus granulosus. Viability of protoscoleces was confirmed by 0.1% aqueous eosin red stain method, where mortality was observed at different concentrations with respect to time. An average anticestodal activity was observed with different endophytic fungal strains, that is, Nigrospora (479 ± 2.9, Colletotrichum (469 ± 25.8, Fusarium (355 ± 14.5, and Chaetomium (332 ± 28.3 showing 64 to 70% protoscolicidal activity, except Pestalotiopsis sp. (581 ± 15.0, which showed promising scolicidal activity up to 97% mortality just within 30 min of incubation. These species showed significant reduction in viability of protoscoleces. This is the first report on the scolicidal activity of endophytic Pestalotiopsis sp. We conclude that ultrastructural changes in protoscoleces were due to endophytic extract suggesting that there may be some bioactive compounds that have selective action on the tegument layer of protoscoleces. As compared with that of standard drug used, endophytic species of Neem plant shows significant anticestodal activity.

  1. Molecular characterization and serodiagnostic potential of a novel dithiol glutaredoxin 1 from Echinococcus granulosus.

    Science.gov (United States)

    Song, Xingju; Yan, Min; Hu, Dandan; Wang, Yu; Wang, Ning; Gu, Xiaobin; Peng, Xuerong; Yang, Guangyou

    2016-08-17

    The larval stage of Echinococcus granulosus is the etiological agent of cystic echinococcosis (CE), which causes serious morbidity and mortality in many areas. There is no reliable method to monitor sheep CE. Here, we characterize E. granulosus glutaredoxin 1 (Eg-Grx1) and report an improved immunodiagnostic method for CE. We cloned and expressed recombinant Eg-Grx1 and generated antibodies. We analyzed the location of the protein in different parasite stages by fluorescence immunohistochemistry, detected the immunogenicity of recombinant Eg-Grx1, and developed an indirect ELISA (iELISA) for CE serodiagnosis. Eg-Grx1 is a classic dithiol Grx with several GSH-binding motifs. Native Eg-Grx1 protein was distributed in the tegument of protoscoleces, the whole germinal layer, and the parenchymatous tissue of adult worms. Recombinant Eg-Grx1 exhibited good immunoreactivity to CE-infected sheep serum. An iELISA using this antigen showed specificity of 64.3 % (9/14) and sensitivity of 1:3200, and the diagnostic accordance rate was 97.9 % (47/48) compared with the results of necropsy. We characterized a novel Grx (Eg-Grx1) from a parasitic helminth and present a comprehensive analysis of the sequence and structure of this protein. The recombinant Eg-Grx1 protein showed good potential serodiagnostic performance, and we established an iELISA method, which may contribute to the surveillance of sheep CE in epidemic areas.

  2. Identification of Albizia lebbeck seed coat chitin-binding vicilins (7S globulins) with high toxicity to the larvae of the bruchid Callosobruchus maculatus

    Energy Technology Data Exchange (ETDEWEB)

    Souza, A.J. [Laboratório de Química e Função de Proteínas e Peptídeos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, RJ (Brazil); Ferreira, A.T.S.; Perales, J.; Beghini, D.G. [Laboratório de Toxinologia, Departamento de Fisiologia e Farmacodinâmica, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, RJ (Brazil); Fernandes, K.V.S.; Xavier-Filho, J.; Venancio, T.M.; Oliveira, A.E.A. [Laboratório de Química e Função de Proteínas e Peptídeos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, RJ (Brazil)

    2011-01-27

    Seed coat is a specialized maternal tissue that interfaces the embryo and the external environment during embryogenesis, dormancy and germination. In addition, it is the first defensive barrier against penetration by pathogens and herbivores. Here we show that Albizia lebbeck seed coat dramatically compromises the oviposition, eclosion and development of the bruchid Callosobruchus maculatus. Dietary supplementation of bruchid larvae with A. lebbeck seed coat flour causes severe weight loss and reduces survival. By means of protein purification, mass spectrometry and bioinformatic analyses, we show that chitinbinding vicilins are the main source of A. lebbeck tegumental toxicity to C. maculatus. At concentrations as low as 0.1%, A. lebbeck vicilins reduce larval mass from 8.1 ± 1.7 (mass of control larvae) to 1.8 ± 0.5 mg, which corresponds to a decrease of 78%. Seed coat toxicity constitutes an efficient defense mechanism, hindering insect predation and preventing embryo damage. We hypothesize that A. lebbeck vicilins are good candidates for the genetic transformation of crop legumes to enhance resistance to bruchid predation.

  3. Dormancy Breaking in Ormosia arborea Seeds

    Directory of Open Access Journals (Sweden)

    Edilma Pereira Gonçalves

    2011-01-01

    Full Text Available Ormosia arborea is a tree species planted in urban areas and used to restore degraded areas. Its seeds are dormant and propagation is difficult. This study compares different dormancy breaking methods and physiological seed quality and seedling production. The seeds were germinated in sand in the laboratory of the Universidade Estadual de Santa Cruz, Ilhéus, Bahia, Brazil. The following dormancy breaking treatments were applied: control (intact seeds, 100°C water immersion; boiling water immersion followed by 24 hours of soaking; scarification with number 100 and number 50 sandpaper opposite from root emergence; sulfuric acid immersion for 1 hour, 50, 45, and 30 minutes. Seed immersion in 100°C and boiling water did not break the dormancy. The study species showed a greater vigor of seedling when its seeds were submitted to treatments associated with tegument rupturing by sandpaper or sulfuric acid. On the other hand, seed scarification with sulfuric acid for 1 hour, 50, 45, and 30 minutes or sandpaper favored seed germination and vigor.

  4. Degradation of rotenone in yam bean seeds ( Pachyrhizus sp.) through food processing.

    Science.gov (United States)

    Catteau, Lucy; Lautié, Emmanuelle; Koné, Oumou; Coppée, Marie; Hell, Kerstin; Pomalegni, Charles Bertrand; Quetin-Leclercq, Joëlle

    2013-11-20

    The purpose of this research is to screen different processes that could potentially decrease or even eliminate rotenone, a toxic isoflavonoid, from Pachyrhizus seeds. Yam bean seeds have very interesting nutritional characteristics, especially their high protein and lipid contents, and could potentially increase food security in under-nourished populations. However, they contain rotenone, a natural molecule previously used as an insecticide inhibiting the respiratory mitochondrial chain. It was also proven to be toxic to mammals as chronic exposure leads to the development of Parkinson-like symptoms in rats. As the thermosensitivity of rotenone had been reported, this study tested different processes (drying, roasting, boiling, frying, alcohol extraction), tegument removal, and traditional Beninese culinary recipes. Rotenone was then quantified in end-products by a validated method, associating microwave extraction, solid phase extraction (SPE), and HPLC-UV. With these processes a rotenone removal of up to 80% was obtained. The most effective methods were the drying and roasting of the seeds and the maceration of their flour in local alcohol. Rotenone degradation and elimination were confirmed by cytotoxic assays, effectively inducing a decrease in sample toxicity.

  5. In vitro organogenesis optimization and plantlet regeneration in Citrus sinensis and C. limonia

    Directory of Open Access Journals (Sweden)

    Almeida Weliton Antonio Bastos de

    2002-01-01

    Full Text Available Exogenous genes can be introduced in plants by genetic transformation techniques. However, an efficient tissue culture system with high rates of plant recovery is necessary for gene introduction. This work aimed to define organogenesis and plant regeneration protocols for sweet orange varieties Natal, Valencia and Hamlin (Citrus sinensis L. Osbeck and Rangpur lime (Citrus limonia L. Osbeck which can be used in plant transformation experiments. Seeds of which teguments were removed, were germinated in vitro and maintained in the dark for three weeks, followed by one week at 16-h photoperiod (40 µmol m-2 s-1 and 27 ± 2°C. Organogenesis induction was done by introducing epicotyl segments in MT medium with 25 g L-1 sucrose and different BAP concentrations. After adventitious bud growth, the shoots were transferred to MT medium with either NAA or IBA (1 mg L-1, or absence of auxin, for rooting. The best results were obtained with 1 mg L-1 BAP for bud induction and 1 mg L-1 IBA for rooting for all three sweet orange cultivars. The use of 0.5-2.5 mg L-1 BAP, followed by 1 mg L-1 IBA were the best growth regulator combinations for bud induction and rooting, respectively, for 'Rangpur' lime. The protocols presented in this work are suitable for associations with genetic transformation experiments for these cultivars.

  6. Sm29, but not Sm22.6 retains its ability to induce a protective immune response in mice previously exposed to a Schistosoma mansoni infection.

    Directory of Open Access Journals (Sweden)

    Clarice Carvalho Alves

    2015-02-01

    Full Text Available BACKGROUND: A vaccine against schistosomiasis would have a great impact in disease elimination. Sm29 and Sm22.6 are two parasite tegument proteins which represent promising antigens to compose a vaccine. These antigens have been associated with resistance to infection and reinfection in individuals living in endemic area for the disease and induced partial protection when evaluated in immunization trials using naïve mice. METHODOLOGY/PRINCIPALS FINDINGS: In this study we evaluated rSm29 and rSm22.6 ability to induce protection in Balb/c mice that had been previously infected with S. mansoni and further treated with Praziquantel. Our results demonstrate that three doses of the vaccine containing rSm29 were necessary to elicit significant protection (26%-48%. Immunization of mice with rSm29 induced a significant production of IL-2, IFN-γ, IL-17, IL-4; significant production of specific antibodies; increased percentage of CD4+ central memory cells in comparison with infected and treated saline group and increased percentage of CD4+ effector memory cells in comparison with naïve Balb/c mice immunized with rSm29. On the other hand, although immunization with Sm22.6 induced a robust immune response, it failed to induce protection. CONCLUSION/SIGNIFICANCE: Our results demonstrate that rSm29 retains its ability to induce protection in previously infected animals, reinforcing its potential as a vaccine candidate.

  7. Histological and ultrastructural localization of antigen B in the metacestode of Taenia solium

    International Nuclear Information System (INIS)

    Laclette, J.P.; Merchant, M.T.; Willms, K.

    1987-01-01

    The morphological localization of antigen B (AgB) in the tissues of the Taenia solium metacestode was studied by immunological and biochemical methods. Indirect immunofluorescence carried out on vibratome sections showed that AgB is widely distributed throughout the tissue. A more intense fluorescence was observed in the tegumentary cytons of the bladder wall and in the lumen of the spiral canal of the invaginated scolex. Ultrastructural analysis of larvae washed in PBS after dissection from meat and then incubated with rabbit antibodies against AgB, followed by peroxidase-labeled goat anti-rabbit IgG, did not exhibit electron-dense material on the external surface. Larvae fixed in glutaraldehyde immediately after dissection and exposed to the immunoperoxidase reagents did exhibit electron-dense material on microtriches, indicating that AgB is only loosely bound to the external surface. Crude extracts of surface-radioiodinated cysticerci analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) contained no labeled proteins with the molecular weight of AgB. Autoradiography of the immunoelectrophoretograms in which the crude extract was confronted with antibodies to AgB demonstrated that this antigen was not labeled, and therefore is not exposed on the tegumentary surface. The results suggest that AgB is synthesized by the tegumentary cytons of the parasite and secreted through the tegumental membrane into the host tissues and the lumen of the spiral canal

  8. Characterization and interactome study of white spot syndrome virus envelope protein VP11.

    Directory of Open Access Journals (Sweden)

    Wang-Jing Liu

    Full Text Available White spot syndrome virus (WSSV is a large enveloped virus. The WSSV viral particle consists of three structural layers that surround its core DNA: an outer envelope, a tegument and a nucleocapsid. Here we characterize the WSSV structural protein VP11 (WSSV394, GenBank accession number AF440570, and use an interactome approach to analyze the possible associations between this protein and an array of other WSSV and host proteins. Temporal transcription analysis showed that vp11 is an early gene. Western blot hybridization of the intact viral particles and fractionation of the viral components, and immunoelectron microscopy showed that VP11 is an envelope protein. Membrane topology software predicted VP11 to be a type of transmembrane protein with a highly hydrophobic transmembrane domain at its N-terminal. Based on an immunofluorescence assay performed on VP11-transfected Sf9 cells and a trypsin digestion analysis of the virion, we conclude that, contrary to topology software prediction, the C-terminal of this protein is in fact inside the virion. Yeast two-hybrid screening combined with co-immunoprecipitation assays found that VP11 directly interacted with at least 12 other WSSV structural proteins as well as itself. An oligomerization assay further showed that VP11 could form dimers. VP11 is also the first reported WSSV structural protein to interact with the major nucleocapsid protein VP664.

  9. Mutational analysis of varicella-zoster virus (VZV) immediate early protein (IE62) subdomains and their importance in viral replication

    Energy Technology Data Exchange (ETDEWEB)

    Khalil, Mohamed I., E-mail: mkhalil2@stanford.edu [Departments of Pediatrics and Microbiology & Immunology, Stan ford University School of Medicine, Stanford, CA (United States); Department of Molecular Biology, National Research Centre, El-Buhouth St., Cairo (Egypt); Che, Xibing; Sung, Phillip; Sommer, Marvin H. [Departments of Pediatrics and Microbiology & Immunology, Stan ford University School of Medicine, Stanford, CA (United States); Hay, John [Department of Microbiology and Immunology, School of Medicine and Biomedical Science, University at Buffalo, Buffalo, NY (United States); Arvin, Ann M. [Departments of Pediatrics and Microbiology & Immunology, Stan ford University School of Medicine, Stanford, CA (United States)

    2016-05-15

    VZV IE62 is an essential, immediate-early, tegument protein and consists of five domains. We generated recombinant viruses carrying mutations in the first three IE62 domains and tested their influence on VZV replication kinetics. The mutations in domain I did not affect replication kinetics while domain II mutations, disrupting the DNA binding and dimerization domain (DBD), were lethal for VZV replication. Mutations in domain III of the nuclear localization signal (NLS) and the two phosphorylation sites S686A/S722A resulted in slower growth in early and late infection respectively and were associated with IE62 accumulation in the cytoplasm and nucleus respectively. This study mapped the functional domains of IE62 in context of viral infection, indicating that DNA binding and dimerization domain is essential for VZV replication. In addition, the correct localization of IE62, whether nuclear or cytoplasmic, at different points in the viral life cycle, is important for normal progression of VZV replication. - Highlights: • Mutation of IE62 domain I did not affect VZV replication in melanoma cells. • IE62 domain II and III are important for VZV replication in melanoma cells. • Mutations of IE62 domain II (DBD) were lethal for virus replication. • Mutations of IE62 NLS and phosphorylation sites inhibited VZV replication. • NLS and S686A/S722A mutations altered localization of IE62 during early and late infection.

  10. The draft genome of the carcinogenic human liver fluke Clonorchis sinensis

    Science.gov (United States)

    2011-01-01

    Background Clonorchis sinensis is a carcinogenic human liver fluke that is widespread in Asian countries. Increasing infection rates of this neglected tropical disease are leading to negative economic and public health consequences in affected regions. Experimental and epidemiological studies have shown a strong association between the incidence of cholangiocarcinoma and the infection rate of C. sinensis. To aid research into this organism, we have sequenced its genome. Results We combined de novo sequencing with computational techniques to provide new information about the biology of this liver fluke. The assembled genome has a total size of 516 Mb with a scaffold N50 length of 42 kb. Approximately 16,000 reliable protein-coding gene models were predicted. Genes for the complete pathways for glycolysis, the Krebs cycle and fatty acid metabolism were found, but key genes involved in fatty acid biosynthesis are missing from the genome, reflecting the parasitic lifestyle of a liver fluke that receives lipids from the bile of its host. We also identified pathogenic molecules that may contribute to liver fluke-induced hepatobiliary diseases. Large proteins such as multifunctional secreted proteases and tegumental proteins were identified as potential targets for the development of drugs and vaccines. Conclusions This study provides valuable genomic information about the human liver fluke C. sinensis and adds to our knowledge on the biology of the parasite. The draft genome will serve as a platform to develop new strategies for parasite control. PMID:22023798

  11. Antibodies against Clonorchis sinensis LDH could cross-react with LDHB localizing on the plasma membrane of human hepatocarcinoma cell SMMC-7721 and induce apoptosis.

    Science.gov (United States)

    Song, Tianzhang; Gan, Wenjia; Chen, Jintao; Huang, Lilin; Yin, Hongling; He, Tailong; Huang, Huaiqiu; Hu, Xuchu

    2016-04-01

    Lactate dehydrogenase (LDH) is a terminal enzyme in anaerobic glycolytic pathway. It widely exists in various organisms and is in charge of converting the glycolysis product pyruvic acid to lactic acid. Most parasites, including Clonorchis sinensis, predominantly depend on glycolysis to provide energy. Bioinformatic analysis predicts that the LDHs from many species have more than one transmembrane region, suggesting that it may be a membrane protein. C. sinensis LDH (CsLDH) has been confirmed as a transmembrane protein mainly located in the tegument. The antibodies against CsLDH can inhibit the worm's energy metabolism, kill the worm, and may have the same effects on human cancer cells. In this study, we cloned and characterized human LDHA (HsLDHA), HsLDHB, and CsLDH. Semi-quantitative real-time RCP showed that HsLDHB only existed in hepatocarcinoma cell SMMC-7721. Confocal microscopy and Western blot experiments revealed that HsLDHB was localized in the plasma membrane of SMMC-7721 cells, and the antibodies against CsLDH could cross-react with it. This cross-reaction could inhibit the enzymatic activity of HsLDHB. The cancer cells co-cultured with anti-CsLDH sera showed a significant decrease in cell proliferation rate and increases in caspase 9 and reactive oxygen species (ROS) levels. Therefore, anti-CsLDH antibodies can induce the apoptosis of cancer cells SMMC-7721 and may serve as a new tool to inhibit tumor.

  12. ESCARIFICAÇÃO ÁCIDA NA SUPERAÇÃO DA DORMÊNCIA DE SEMENTES DE PAU FERRO (Caesalpinea ferrea Mart.ex Tu. var. leiostachya Benth.

    Directory of Open Access Journals (Sweden)

    Edna Ursulino Alves

    2009-01-01

    Full Text Available Seeds of Caesalpinea ferrea Mart. ex Tu. var. leiostachya Benth. present low and irregular germination due to dormancy caused by the impermeability of the tegument. With the purpose to determine an efficient method to accelerate and uniformize seed germination, they were submitted to different immersion periods (0, 5, 10, 15, 20, 25 and 30 minutes in sulfuric acid (95-98%, influence emergency and vigor (first count, speed index, medium time and relative frequency of emergency, length and dry matter of the seedlings. The experiment was carried out under greenhouse conditions with an experimental design of completely randomized, with seven treatments and four replications. The pre-conditioning of the seeds with immersion in concentrated sulfuric acid was efficient to overcoming seed dormancy, by increasing the percentage and speed emergency, the first count of emergency, length and dry matter of seedlings and reduction in the medium time for emergency. The efficiency of this chemical treatment with concentrated sulfuric depends on the immersion time, and 10 to 20 minutes were the most adequate to provide larger emergency uniformity percentages and vigor.

  13. The survival of monogenean (platyhelminth) parasites on fish skin.

    Science.gov (United States)

    Kearn, G C

    1999-01-01

    This review deals with the problems faced by those monogenean (platyhelminth) parasites that attach themselves to fish skin. The structure of the skin and the ways in which the posterior hook-bearing haptor achieves virtually permanent attachment to the skin are considered. Small marginal hooklets are specialized for attachment to superficial host epidermal cells, finding anchorage in the terminal web of keratinous tonofilaments, while large hooks (hamuli) may penetrate into and lodge in the collagenous dermis. The complementary roles of suction and sticky secretions in haptor attachment and the role of the pharynx in temporary attachment during feeding are also considered. During leech-like locomotion the haptor is briefly detached and, at this critical time, the anterior end is strongly fixed to the wet, current-swept and possibly slimy skin by a sticky secretion. This secretion is deployed on paired pads or discs, the latter sometimes backed up by suction. After attachment by the haptor is re-established, the special tegument covering the anterior adhesive areas may be instrumental in their instant release. The role of fish skin in the phenomenon of host specificity and in the generation of a defensive response against monogeneans is considered and site-specificity of parasites on the host's body is discussed. Possible selection pressures exerted by predatory 'cleaner' organisms are briefly evaluated.

  14. Effects of dietary antioxidant of tomato extract and lycopene on Carassius auratus and Xiphophorus maculatus

    Directory of Open Access Journals (Sweden)

    Cynthia Montoya M.

    2014-06-01

    Full Text Available Objective. Evaluate the effect on tegument pigmentation, survival, growth and antioxidant capacity in diets supplemented with tomato extract and lycopene as additives in experimental feed for Carassius auratus and Xiphophorus maculatus. Materials and methods. The additives were added in different concentrations to a basic diet. We performed beginning and an ending biometrics for 100% of the population in each bioassay. The growth and survival of organisms were evaluated. The antioxidant capacity was analyzed by ABTS assay, both in the tomato extract sample as well as in foods used in different bioassays. The concentration of lycopene was determined in food and liver and muscle samples of fish fed with it. Acquired pigmentation of fish was assessed through photographs analyzed with Adobe Photoshop®. The results were evaluated by analysis of variance, and when differences were found (p0.05 on pigmentation and growth of the organisms under the established experimental conditions was obtained. Significant differences in antioxidant capacity (p<0.05 were obtained in foods with added lycopene. Conclusions. The inclusion of lycopene or tomato extract in food for the organisms used is not recommended to improve pigmentation, but further studies are needed to demonstrate antioxidant effect.

  15. Anthelmintic effect of thymol and thymol acetate on sheep gastrointestinal nematodes and their toxicity in mice

    Directory of Open Access Journals (Sweden)

    Weibson Paz Pinheiro André

    Full Text Available Abstract Thymol is a monoterpene and acetylation form of this compound can reduce the toxicity and enhance its biological effects. The objective of this study was to evaluate the effect of thymol and thymol acetate (TA on egg, larva and adult Haemonchus contortus and the cuticular changes, acute toxicity in mice and the efficacy on sheep gastrointestinal nematodes. In vitro tests results were analyzed by analysis of variance (ANOVA and followed by comparison with Tukey test or Bonferroni. The efficacy of in vivo test was calculated by the BootStreet program. In the egg hatch test (EHT, thymol (0.5 mg/mL and TA (4 mg/mL inhibited larval hatching by 98% and 67.1%, respectively. Thymol and TA (8 mg/mL inhibited 100% of larval development. Thymol and TA (800 µg/mL reduced the motility of adult worms, by 100% and 83.4%, respectively. Thymol caused cuticular changes in adult worm teguments. In the acute toxicity test, the LD50 of thymol and TA were 1,350.9 mg/kg and 4,144.4 mg/kg, respectively. Thymol and TA reduced sheep egg count per gram of faeces (epg by 59.8% and 76.2%, respectively. In in vitro tests thymol presented better anthelmintic activity than TA. However TA was less toxic and in in vivo test efficacy was similar.

  16. Identification of Albizia lebbeck seed coat chitin-binding vicilins (7S globulins) with high toxicity to the larvae of the bruchid Callosobruchus maculatus

    International Nuclear Information System (INIS)

    Souza, A.J.; Ferreira, A.T.S.; Perales, J.; Beghini, D.G.; Fernandes, K.V.S.; Xavier-Filho, J.; Venancio, T.M.; Oliveira, A.E.A.

    2011-01-01

    Seed coat is a specialized maternal tissue that interfaces the embryo and the external environment during embryogenesis, dormancy and germination. In addition, it is the first defensive barrier against penetration by pathogens and herbivores. Here we show that Albizia lebbeck seed coat dramatically compromises the oviposition, eclosion and development of the bruchid Callosobruchus maculatus. Dietary supplementation of bruchid larvae with A. lebbeck seed coat flour causes severe weight loss and reduces survival. By means of protein purification, mass spectrometry and bioinformatic analyses, we show that chitinbinding vicilins are the main source of A. lebbeck tegumental toxicity to C. maculatus. At concentrations as low as 0.1%, A. lebbeck vicilins reduce larval mass from 8.1 ± 1.7 (mass of control larvae) to 1.8 ± 0.5 mg, which corresponds to a decrease of 78%. Seed coat toxicity constitutes an efficient defense mechanism, hindering insect predation and preventing embryo damage. We hypothesize that A. lebbeck vicilins are good candidates for the genetic transformation of crop legumes to enhance resistance to bruchid predation

  17. Identification of Albizia lebbeck seed coat chitin-binding vicilins (7S globulins) with high toxicity to the larvae of the bruchid Callosobruchus maculatus.

    Science.gov (United States)

    Souza, A J; Ferreira, A T S; Perales, J; Beghini, D G; Fernandes, K V S; Xavier-Filho, J; Venancio, T M; Oliveira, A E A

    2012-02-01

    Seed coat is a specialized maternal tissue that interfaces the embryo and the external environment during embryogenesis, dormancy and germination. In addition, it is the first defensive barrier against penetration by pathogens and herbivores. Here we show that Albizia lebbeck seed coat dramatically compromises the oviposition, eclosion and development of the bruchid Callosobruchus maculatus. Dietary supplementation of bruchid larvae with A. lebbeck seed coat flour causes severe weight loss and reduces survival. By means of protein purification, mass spectrometry and bioinformatic analyses, we show that chitin-binding vicilins are the main source of A. lebbeck tegumental toxicity to C. maculatus. At concentrations as low as 0.1%, A. lebbeck vicilins reduce larval mass from 8.1 ± 1.7 (mass of control larvae) to 1.8 ± 0.5 mg, which corresponds to a decrease of 78%. Seed coat toxicity constitutes an efficient defense mechanism, hindering insect predation and preventing embryo damage. We hypothesize that A. lebbeck vicilins are good candidates for the genetic transformation of crop legumes to enhance resistance to bruchid predation.

  18. Proteomic characterization of murid herpesvirus 4 extracellular virions.

    Directory of Open Access Journals (Sweden)

    Sarah Vidick

    Full Text Available Gammaherpesvirinae, such as the human Epstein-Barr virus (EBV and the Kaposi's sarcoma associated herpesvirus (KSHV are highly prevalent pathogens that have been associated with several neoplastic diseases. As EBV and KSHV are host-range specific and replicate poorly in vitro, animal counterparts such as Murid herpesvirus-4 (MuHV-4 have been widely used as models. In this study, we used MuHV-4 in order to improve the knowledge about proteins that compose gammaherpesviruses virions. To this end, MuHV-4 extracellular virions were isolated and structural proteins were identified using liquid chromatography tandem mass spectrometry-based proteomic approaches. These analyses allowed the identification of 31 structural proteins encoded by the MuHV-4 genome which were classified as capsid (8, envelope (9, tegument (13 and unclassified (1 structural proteins. In addition, we estimated the relative abundance of the identified proteins in MuHV-4 virions by using exponentially modified protein abundance index analyses. In parallel, several host proteins were found in purified MuHV-4 virions including Annexin A2. Although Annexin A2 has previously been detected in different virions from various families, its role in the virion remains controversial. Interestingly, despite its relatively high abundance in virions, Annexin A2 was not essential for the growth of MuHV-4 in vitro. Altogether, these results extend previous work aimed at determining the composition of gammaherpesvirus virions and provide novel insights for understanding MuHV-4 biology.

  19. Parasite enzymes as a tool to investigate immune responses

    Directory of Open Access Journals (Sweden)

    Italo M. Cesari

    1992-01-01

    Full Text Available Previous evidences reported by us and by other authors revealed the presence of IgG in sera of Schistosoma mansoni-infected patients to immunodominant antigens which are enzymes. Besides their immunological interest as possible inductors of protection, several of these enzume antigens might be also intersting markers of infection in antibody-detecting immunocapture assays which use the intrinsic catalytic property of these antigens. It was thus thought important to define some enzymatic and immunological characteristics of these molecules to better exploit their use as antigens. Four different enzymes from adult worms were partially characterized in their biochemical properties and susceptibility to react with antibodies of infected patients, namely alkaline phosphatase (AKP, Mg*+, pH 9.5, type I phosphodiesterase (PDE, pH 9.5, cysteine proteinase (CP, dithiothreitol, pH 5.5 and N-acetyl-ß-D-glucosaminidase (NAG, pH 5.5. The AKP and PDE are distinct tegumental membrane-bound enzymes whereas CP and NAG are soluble acid enzymes. Antibodies in infected human sera differed in their capacity to react with and to inhibit these enzyme antigens. Possibly, the specificity of the antibodies related to the extent of homology between the parasite and the host enzyme might be in part responsible for the above differences. The results are also discussed in view of the possible functional importance of these enzymes.

  20. Morphology and surface topography of the schistosome Bivitellobilharzia nairi from the Asian elephant (Elephas maximus maximus) in Sri Lanka.

    Science.gov (United States)

    Rajapakse, R P V J; Iwagami, M; Wickramasinghe, S; Walker, S M; Agatsuma, T

    2013-09-01

    Bivitellobilharzia nairi was first recorded from an Indian elephant (Elephas maximus) in Berlin. Infections with this parasite have become increasingly important in E. maximus maximus populations in Sri Lanka. The present work is the first morphological description of this schistosome from Sri Lanka. A number of adult worms were recovered from a dead Asian elephant near the elephant orphanage, Pinnawala, in Sri Lanka. The observed clinical features of the infected elephant included emaciation, subventral oedema and anaemia. Post-mortem results indicated that the liver was enlarged and adult schistosomes were found in the blood vessels of the liver parenchyma. The total number of worms recovered from a portion of the liver was 129,870, which is an average of 22 worms per 100 g of liver. The present study uses both light microscopic and scanning electron microscope (SEM) techniques for the morphological and topographical characterization of this parasite and to permit comparison with other species of schistosomes. Morphologically, these worms correspond very well to the description of B. nairi by Dutt & Srivastava (1955). Moreover, it is clear that B. nairi is a distinctive species easily differentiated from other schistosomes. The SEM study of the tegument of male worms shows that the surface of B. nairi is smoother than in other schistosomes.

  1. Immunological and biochemical studies of fascioliasis in goats and cattle

    International Nuclear Information System (INIS)

    Reddington, J.J.

    1985-01-01

    Using the goat as a susceptible host and cattle as a resistant species to Fasciola hepatica infections, the humoral response of these animals to the surface of the newly excysted juvenile (NEJ) fluke was examined. Tegumental proteins of the NEJ were labeled with 125 I by lactoperoxidase and analyzed after immunoprecipitation using a double antibody system. In addition, a comparison was made between the infected sera's capacity to immunoprecipitate surface antigens and their in vitro cytotoxic activity against the NEJ. In both goats and cattle the levels of NEJ surface antigens precipitated increased during the first 4 weeks PI. The peak immunoprecipitation of NEJ surface antigens by cattle sera (58%) was significantly higher than that of infected goat sera (33%). Immunoprecipitation of the available radiolabeled NEJ surface proteins by the infected cattle sera remained consistently higher than goat sera until the 16th week PI. The cytotoxic effects of these same caprine sera on NEJs in vitro was limited, while the cytotoxicity of the infected bovine sera closely approximated the sera's ability to precipitate NEJ surface antigens. There was also a qualitative difference between the species in their recognition of 35 S and 125 I radiolabeled NEJ surface antigens. Uninfected goat or cattle sera failed to precipitate any 125 I or 35 S-labeled surface proteins

  2. Immunodetection of Fasciola gigantica Circulating Antigen in Sera of Infected Individuals for Laboratory Diagnosis of Human Fascioliasis

    Science.gov (United States)

    Attallah, Abdelfattah M.; Bughdadi, Faisal A.; El-Shazly, Atef M.

    2013-01-01

    Currently, the laboratory diagnosis of human fascioliasis is based on the parasitological examination of parasite eggs in stool specimens and serological detection of specific antibodies in serum samples, which are often unreliable diagnostic approaches. Ideally, a sensitive and specific diagnostic test for Fasciola infection should be based on the detection of circulating Fasciola antigen, which implies active infection. Here, a 27-kDa-molecular-mass antigen was identified in a Fasciola gigantica adult worm antigen preparation, excretory-secretory products, and sera from F. gigantica-infected individuals, and it was not detected in antigenic extracts of other parasites and sera from noninfected individuals. The target antigen was isolated and partially characterized as a protein. Immunoperoxidase staining located the target epitope within teguments and guts of F. gigantica adult worms. The performance characteristics of a newly developed enzyme-linked immunosorbent assay (ELISA) based on F. gigantica circulating antigen detection in serum (FgCA-27 ELISA) were investigated using sera of 120 parasitologically diagnosed F. gigantica-infected individuals and 80 noninfected individuals. The area under the receiving operating characteristic (ROC) curve (AUC) for ELISA was significantly high (AUC = 0.961, P 93%), and a significant correlation (r = 0.715, P fascioliasis. PMID:23945158

  3. Immunodetection of Fasciola gigantica circulating antigen in sera of infected individuals for laboratory diagnosis of human fascioliasis.

    Science.gov (United States)

    Attallah, Abdelfattah M; Bughdadi, Faisal A; El-Shazly, Atef M; Ismail, Hisham

    2013-10-01

    Currently, the laboratory diagnosis of human fascioliasis is based on the parasitological examination of parasite eggs in stool specimens and serological detection of specific antibodies in serum samples, which are often unreliable diagnostic approaches. Ideally, a sensitive and specific diagnostic test for Fasciola infection should be based on the detection of circulating Fasciola antigen, which implies active infection. Here, a 27-kDa-molecular-mass antigen was identified in a Fasciola gigantica adult worm antigen preparation, excretory-secretory products, and sera from F. gigantica-infected individuals, and it was not detected in antigenic extracts of other parasites and sera from noninfected individuals. The target antigen was isolated and partially characterized as a protein. Immunoperoxidase staining located the target epitope within teguments and guts of F. gigantica adult worms. The performance characteristics of a newly developed enzyme-linked immunosorbent assay (ELISA) based on F. gigantica circulating antigen detection in serum (FgCA-27 ELISA) were investigated using sera of 120 parasitologically diagnosed F. gigantica-infected individuals and 80 noninfected individuals. The area under the receiving operating characteristic (ROC) curve (AUC) for ELISA was significantly high (AUC = 0.961, P 93%), and a significant correlation (r = 0.715, P fascioliasis.

  4. Using composting for control seed germination of invasive plant (water hyacinth) in Extremadura (Spain)

    Science.gov (United States)

    Labrador, Juana; Gordillo, Judit; Ruiz, Trinidad; Albano, Eva; Moreno, Marta M.

    2016-04-01

    The biotransformation of the invasive water hyacinth (Eichhornia crassipes) by composting has been showed as a viable alternative to offset the economic cost of eliminating an invasive plant giving a value to the by-product; however, as result of the propagative plant capacity, it was necessary to check if the composting process could eliminate the germination seed rate. Despite the high temperatures and the biochemical biotransformation processes of the composting components, in the case of seed water hyacinth, with a recovery rate of 100%, damage was observed in some parts of the seed anatomy such as in the outer teguments; however, other parts of the seed coat and the endosperm maintained their integrity. A microscopic analysis revealed that the embryo was noticeable and this was supported by the rate of seed germination observed (3.5 ± 0.96%). The results indicate that the use of water hyacinth for compost production is not completely safe from an environmental perspective. Keywords: Eichhornia crassipes, water hyacinth, invasive plant, seed anatomy, seed germination rate, compost. References: Ruiz T., Martín de Rodrigo E., Lorenzo G., Albano E., Morán R., Sánchez J.M. 2008. The Water Hyacinth, Eichhornia crassipes: an invasive plant in the Guadiana River Basin (Spain). Aquatic Invasions Volume 3, Issue 1:42-53.

  5. Advances in the processing of policromat images as diagnostic method to determine white spot syndrome virus in white shrimp (Litopenaeus vannamei)

    Science.gov (United States)

    Chavez-Sanchez, Cristina M.; Alvarez-Borrego, Josue; Montoya-Rodriguez, L.; Garcia-Gasca, A.; Fajer Avila, Emma J.; Pacheco-Marges, R.

    2004-10-01

    White spot syndrome (WSSV) is a viral disease which affects many crustacean species including commercial shrimps. Adequate, precise and quick methods to diagnose on time the presence of the disease in order to apply different strategies to avoid the dispersion and to reduce mortalities is necessary. Histopathology is an important diagnostic method. However, histopathology has the problem that requires time to prepare the histological slides and time to arrive to some diagnosis because this depend on the nature of the tissues, the pathogen(s) to find, the number of organisms, number of slides to analyze and the skill of the technician. This paper try to demonstrate the sensibility of one digital system of processing and recognition of images using color correlation with phase filters, to identify inclusion bodies of WSSV. Infected tissues were processed to obtain histological slides and to verify that the inclusion bodies observed were of WSV, in situ hybridization were carried out. The sensibility results of the recognition of the inclusion bodies of WSSV with the color correlation program was 86.1%. The highest percentage of recognition was in nervous system and tegument glands with 100%. The values in the stomach epithelium and heart tissue was 78.45% of recognition. Tissues with the lowest recognition values were lymphoid organ and hematopoietic tissue. It is necessary further studies to increase the sensibility and to obtain the specificity.

  6. Identification of Albizia lebbeck seed coat chitin-binding vicilins (7S globulins with high toxicity to the larvae of the bruchid Callosobruchus maculatus

    Directory of Open Access Journals (Sweden)

    A.J. Souza

    2012-02-01

    Full Text Available Seed coat is a specialized maternal tissue that interfaces the embryo and the external environment during embryogenesis, dormancy and germination. In addition, it is the first defensive barrier against penetration by pathogens and herbivores. Here we show that Albizia lebbeck seed coat dramatically compromises the oviposition, eclosion and development of the bruchid Callosobruchus maculatus. Dietary supplementation of bruchid larvae with A. lebbeck seed coat flour causes severe weight loss and reduces survival. By means of protein purification, mass spectrometry and bioinformatic analyses, we show that chitin-binding vicilins are the main source of A. lebbeck tegumental toxicity to C. maculatus. At concentrations as low as 0.1%, A. lebbeck vicilins reduce larval mass from 8.1 ± 1.7 (mass of control larvae to 1.8 ± 0.5 mg, which corresponds to a decrease of 78%. Seed coat toxicity constitutes an efficient defense mechanism, hindering insect predation and preventing embryo damage. We hypothesize that A. lebbeck vicilins are good candidates for the genetic transformation of crop legumes to enhance resistance to bruchid predation.

  7. Comparative Evaluation of Anthelmintic Activity of Edible and Ornamental Pomegranate Ethanolic Extracts against Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    Doaa A. Yones

    2016-01-01

    Full Text Available Due to the development of praziquantel (PZQ schistosomes resistant strains, the discovery of new antischistosomal agents is of high priority in research. This work reported the in vitro and in vivo effects of the edible and ornamental pomegranate extracts against Schistosoma mansoni. Leaves and stem bark ethanolic extracts of both dried pomegranates were prepared at 100, 300, and 500 μg/mL for in vitro and 600 and 800 mg/kg for in vivo. Adult worms Schistosoma mansoni in RPMI-1640 medium for in vitro and S. mansoni infected mice for in vivo tests were obtained from Theodor Bilharz Research Institute, Cairo, Egypt. In vitro activity was manifested by significant coupled worms separation, reduction of motor activity, lethality, and ultrastructural tegumental alterations in adult worms. In vivo activity was manifested revealed by significant reduction of hepatic granulomas number and diameter, decreased number of bilharzial eggs in liver tissues, lowered liver inflammatory infiltration, decreased hepatic fibrosis, and inducible nitric oxide synthase (iNOS expression. Ethanolic stem bark extract of edible pomegranate exhibited highest antischistosomal activities both in vitro and in vivo. Therefore, pomegranate showed a good potential to be used as a promising new candidate for the development of new schistosomicidal agents.

  8. Foliar additional nutrition in the fruit growing field

    International Nuclear Information System (INIS)

    Soare, M.; Borlan, Z.; Gavriluta, I.; Budoi, G.; Marinca, C.; Bandu, G.G.

    1999-01-01

    This paper presents data concerning the influence of foliar application of some types of complex foliar fertilizers under the conditions of SCPP Caransebes (Caras-Severin district). The composition of these nutrients fulfils the nutritional needs of the fruit growing species on the fruit yield obtained in orchards. The application of different types of foliar nutrients on plants teguments resulted in some significant yield increases for the two species that were studied: apple and plum tree. The novelty of this paper is represented by the apparent degrees of productive use in yields of the macro and micronutrients from foliar fertilizers, as well as the productive use degrees of nutrients present in soil and of the nutrients applied in soil (we took into account the mean values for the studied years). The apparent degrees of productive use (in yield increases) of the nutrients from complex foliar fertilizers applied on apple-trees and plum-trees generally exceed 100 %. They determine high levels of productive use of the nutrients from soil and foliar nutrients applied. The experimental data emphasize the ecological protection effect of the supplementary foliar fertilization for the yield stimulation especially on soils with light texture and sloping soils. Refs. 6 (author)

  9. Hoopoes color their eggs with antimicrobial uropygial secretions

    Science.gov (United States)

    Soler, Juan J.; Martín-Vivaldi, M.; Peralta-Sánchez, J. M.; Arco, L.; Juárez-García-Pelayo, N.

    2014-09-01

    Uropygial gland secretions are used as cosmetics by some species of birds to color and enhance properties of feathers and teguments, which may signal individual quality. Uropygial secretions also reach eggshells during incubation and, therefore, may influence the coloration of birds' eggs, a trait that has attracted the attention of evolutionary biologists for more than one century. The color of hoopoe eggs typically changes along incubation, from bluish-gray to greenish-brown. Here, we test experimentally the hypothesis that dark uropygial secretion of females is responsible for such drastic color change. Moreover, since uropygial secretion of hoopoes has antimicrobial properties, we also explore the association between color and antimicrobial activity of the uropygial secretion of females. We found that eggs stayed bluish-gray in nests where female access to the uropygial secretion was experimentally blocked. Furthermore, experimental eggs that were maintained in incubators and manually smeared with uropygial secretion experienced similar color changes that naturally incubated eggs did, while control eggs that were not in contact with the secretions did not experience such color changes. All these results strongly support the hypothesis that female hoopoes use their uropygial gland secretion to color the eggs. Moreover, saturation of the uropygial secretion was associated with antimicrobial activity against Bacillus licheniformis. Given the known antimicrobial potential of uropygial secretions of birds, this finding opens the possibility that in scenarios of sexual selection, hoopoes in particular and birds in general signal antimicrobial properties of their uropygial secretion by mean of changes in egg coloration along incubation.

  10. Lethal dose and clinical signs of Aeromonas hydrophila in Arapaima gigas (Arapaimidae), the giant fish from Amazon.

    Science.gov (United States)

    Dias, Marcia K R; Sampaio, Luciana S; Proietti-Junior, Aldo A; Yoshioka, Eliane T O; Rodrigues, Dália P; Rodriguez, Anselmo F R; Ribeiro, Ricardo A; Faria, Fernando S E D V; Ozório, Rodrigo O A; Tavares-Dias, Marcos

    2016-05-30

    Aeromonas hydrophila is causing substantial economic losses in world aquaculture. This study determined the tolerance limit (LD50-96h) of A. hydrophila in Arapaima gigas, and also investigated the clinical signs after intradermal inoculation. Arapaima gigas fingerlings were inoculated intraperitoneally with 0 (control), 1.0×10(5), 1.0×10(6), 1.0×10(7), 1.0×10(9) and 1.0×10(10)CFU/mL of A. hydrophila for the determination of LD50-96h, which was 1.8×10(8)CFU/mL. In another trial with intradermal inoculation of 1.8×10(8)CFU/mL A. hydrophila, there was a 91.6% of mortality between 8 and 23h, and several clinical signs were found. As follows: depigmentation in the tegument, lesions in the tail and fins, loss of balance, reduction of respiratory movements, hemorrhagic foci, necrotic hemorrhages in the kidney, liver and swim bladder, splenomegaly, ascites in the abdominal cavity and hyperemia, enlargement of the gall bladder, among other clinical signs observed. The results showed that A. gigas has a relative tolerance to A. hydrophila when compared to other Neotropical fish species. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Pectinase production by Penicillium viridicatum RFC3 by solid state fermentation using agricultural wastes and agro-industrial by-products

    Directory of Open Access Journals (Sweden)

    Silva Dênis

    2002-01-01

    Full Text Available Pectin lyase and polygalacturonase production by newly isolated Penicillium viridicatum strain Rfc3 was carried out by means of solid state fermentation using orange bagasse, corn tegument, wheat bran and mango and banana peels as carbon sources. The maximal activity value of polygalacturonase (Pg (30U.g-1 was obtained using wheat bran as carbon source while maximal pectin lyase (Pl (2000 U.g-1 activity value was obtained in medium composed of orange bagasse. Mixtures of banana or mango peels with sugar cane bagasse resulted in increased Pg and Pl production compared to fermentations in which this residue was not used. The mixture of orange bagasse and wheat bran (50% increased the production of Pg and Pl to 55 U.g-1 and 3540 U.g -1 respectively. Fractions of Pg and Pl, isolated by gel filtration in Sephadex G50, presented optimum activity at pH 5.0 and 10.5 respectively. Maximal activity of Pg and Pl fractions was determined at 55ºC and 50ºC respectively. Pg was stable in neutral pH range and at 40ºC whereas Pl was stable in acidic pH and at 35ºC, for 1 h.

  12. VP22 herpes simplex virus protein can transduce proteins into stem cells

    International Nuclear Information System (INIS)

    Gabanyi, I.; Lojudice, F.H.; Kossugue, P.M.; Rebelato, E.; Demasi, M.A.; Sogayar, M.C.

    2013-01-01

    The type I herpes simplex virus VP22 tegument protein is abundant and well known for its ability to translocate proteins from one cell to the other. In spite of some reports questioning its ability to translocate proteins by attributing the results observed to fixation artifacts or simple attachment to the cell membrane, VP22 has been used to deliver several proteins into different cell types, triggering the expected cell response. However, the question of the ability of VP22 to enter stem cells has not been addressed. We investigated whether VP22 could be used as a tool to be applied in stem cell research and differentiation due to its capacity to internalize other proteins without altering the cell genome. We generated a VP22.eGFP construct to evaluate whether VP22 could be internalized and carry another protein with it into two different types of stem cells, namely adult human dental pulp stem cells and mouse embryonic stem cells. We generated a VP22.eGFP fusion protein and demonstrated that, in fact, it enters stem cells. Therefore, this system may be used as a tool to deliver various proteins into stem cells, allowing stem cell research, differentiation and the generation of induced pluripotent stem cells in the absence of genome alterations

  13. From genomes to vaccines via the proteome

    Directory of Open Access Journals (Sweden)

    R Alan Wilson

    2004-08-01

    Full Text Available An effective vaccine against schistosomiasis mansoni would be a valuable control tool and the high levels of protection elicited in rodents and primates by radiation-attenuated cercariae provide proof of principle. A major obstacle to vaccine development is the difficulty of identifying the antigens that mediate protection, not least because of the size of the genome at 280Mb DNA encoding 14,000 to 20,000 genes. The technologies collectively called proteomics, including 2D electrophoresis, liquid chromatography and mass spectrometry, now permit any protein to be identified provided there is extensive DNA data, and preferably a genome sequence. Applied to soluble (cytosolic proteins from schistosomes, proteomics reveals the great similarity in composition between life cycle stages, with several WHO vaccine candidates amongst the most abundant constituents. The proteomic approach has been successfully applied to identify the secretions used by cercaria to penetrate host skin, the gut secretions of adult worms and the proteins exposed on the tegument surface. Soluble proteins can also be separated by 2D electrophoresis before western blotting to identify the full range of antigenic targets present in a parasite preparation. The next step is to discover which target proteins represent the weak points in the worm's defences.

  14. Solution Structure, Membrane Interactions, and Protein Binding Partners of the Tetraspanin Sm-TSP-2, a Vaccine Antigen from the Human Blood Fluke Schistosoma mansoni*

    Science.gov (United States)

    Jia, Xinying; Schulte, Leigh; Loukas, Alex; Pickering, Darren; Pearson, Mark; Mobli, Mehdi; Jones, Alun; Rosengren, Karl J.; Daly, Norelle L.; Gobert, Geoffrey N.; Jones, Malcolm K.; Craik, David J.; Mulvenna, Jason

    2014-01-01

    The tetraspanins (TSPs) are a family of integral membrane proteins that are ubiquitously expressed at the surface of eukaryotic cells. TSPs mediate a range of processes at the surface of the plasma membrane by providing a scaffold for the assembly of protein complexes known as tetraspanin-enriched microdomains (TEMs). We report here the structure of the surface-exposed EC2 domain from Sm-TSP-2, a TSP from Schistosoma mansoni and one of the better prospects for the development of a vaccine against schistosomiasis. This is the first solution structure of this domain, and our investigations of its interactions with lipid micelles provide a general model for interactions between TSPs, membranes, and other proteins. Using chemical cross-linking, eight potential protein constituents of Sm-TSP-2-mediated TEMs were also identified. These include proteins important for membrane maintenance and repair, providing further evidence for the functional role of Sm-TSP-2- and Sm-TSP-2-mediated TEMs. The identification of calpain, Sm29, and fructose-bisphosphate aldolase, themselves potential vaccine antigens, suggests that the Sm-TSP-2-mediated TEMs could be disrupted via multiple targets. The identification of further Sm-TSP-2-mediated TEM proteins increases the available candidates for multiplex vaccines and/or novel drugs targeting TEMs in the schistosome tegument. PMID:24429291

  15. Solution structure, membrane interactions, and protein binding partners of the tetraspanin Sm-TSP-2, a vaccine antigen from the human blood fluke Schistosoma mansoni.

    Science.gov (United States)

    Jia, Xinying; Schulte, Leigh; Loukas, Alex; Pickering, Darren; Pearson, Mark; Mobli, Mehdi; Jones, Alun; Rosengren, Karl J; Daly, Norelle L; Gobert, Geoffrey N; Jones, Malcolm K; Craik, David J; Mulvenna, Jason

    2014-03-07

    The tetraspanins (TSPs) are a family of integral membrane proteins that are ubiquitously expressed at the surface of eukaryotic cells. TSPs mediate a range of processes at the surface of the plasma membrane by providing a scaffold for the assembly of protein complexes known as tetraspanin-enriched microdomains (TEMs). We report here the structure of the surface-exposed EC2 domain from Sm-TSP-2, a TSP from Schistosoma mansoni and one of the better prospects for the development of a vaccine against schistosomiasis. This is the first solution structure of this domain, and our investigations of its interactions with lipid micelles provide a general model for interactions between TSPs, membranes, and other proteins. Using chemical cross-linking, eight potential protein constituents of Sm-TSP-2-mediated TEMs were also identified. These include proteins important for membrane maintenance and repair, providing further evidence for the functional role of Sm-TSP-2- and Sm-TSP-2-mediated TEMs. The identification of calpain, Sm29, and fructose-bisphosphate aldolase, themselves potential vaccine antigens, suggests that the Sm-TSP-2-mediated TEMs could be disrupted via multiple targets. The identification of further Sm-TSP-2-mediated TEM proteins increases the available candidates for multiplex vaccines and/or novel drugs targeting TEMs in the schistosome tegument.

  16. Morphological effects on helminth parasites caused by herbicide under experimental conditions

    Directory of Open Access Journals (Sweden)

    Tainá Carneiro de Castro Monte

    2018-02-01

    Full Text Available Abstract Helminth parasites have been studied as potential accumulators for different pollutants. Echinostoma paraensei is a foodborne trematode whose vertebrate host, the rodent Nectomys squamipes, is naturally exposed to environmental pesticides. However, little information exists regarding the pesticide’s effects on helminths. This study investigated the morphological effects on the trematode, E. paraensei, after experimental Roundup® herbicide exposure, in concentrations below those recommended for agricultural use. After two hours of exposure, scanning electron microscopy (SEM showed changes to the tegument, such as furrowing, shrinkage, peeling, spines loss on the peristomic collar, and histopathological evidence of altered cells in the cecum and acinus vitelline glands with vacuoles and structural changes to the muscular layers. Glycidic content was decreased, primarily in the connective tissue. As E. paraensei is an intestinal parasite of the semi-aquatic wild rodent, N. squamipes, it is predisposed to pesticide exposure resulting from agricultural practices. Therefore, we emphasize the need to evaluate its impact on helminth parasites, due to their pivotal role in regulating host populations.

  17. Mutation induction of pleurotus ferulae by low-energy N+ ion implantation and characters of the selected mutant

    International Nuclear Information System (INIS)

    Chen Henglei; Wan Honggui; Zhang Jun; Zeng Xianxian

    2008-01-01

    In order to obtain Pleurotus ferulae with high temperature tolerance, mycelium mono-cells of wild type strain ACK was treated by nitrogen ion (5-30 keV, 1.5x10 15 -1.5x10 16 cm -2 ) implantation, and mutant CGMCC1762 was selected through auxotrophy screening method, which was Lys, VB6 auxotrophy stress with high temperature. We found that during riper period the surface layer mycelium of the mutant was not aging neither grew tegument even above 30 degree C. The mycelium endurable temperature of the mutant was increased 7 degree C compared with that of the wild type strain. The fruiting bodies growth temperature of the mutant was 16-20 degree C in daytime and was 6-12 degree C at night. The highest growth temperature of fruiting bodies of the mutant was increased by 5 degree C than that of original strain. Through three generation investigation, we found that the mutant CGMCC1762 was stable with high temperature tolerance. (authors)

  18. Mutational analysis of varicella-zoster virus (VZV) immediate early protein (IE62) subdomains and their importance in viral replication

    International Nuclear Information System (INIS)

    Khalil, Mohamed I.; Che, Xibing; Sung, Phillip; Sommer, Marvin H.; Hay, John; Arvin, Ann M.

    2016-01-01

    VZV IE62 is an essential, immediate-early, tegument protein and consists of five domains. We generated recombinant viruses carrying mutations in the first three IE62 domains and tested their influence on VZV replication kinetics. The mutations in domain I did not affect replication kinetics while domain II mutations, disrupting the DNA binding and dimerization domain (DBD), were lethal for VZV replication. Mutations in domain III of the nuclear localization signal (NLS) and the two phosphorylation sites S686A/S722A resulted in slower growth in early and late infection respectively and were associated with IE62 accumulation in the cytoplasm and nucleus respectively. This study mapped the functional domains of IE62 in context of viral infection, indicating that DNA binding and dimerization domain is essential for VZV replication. In addition, the correct localization of IE62, whether nuclear or cytoplasmic, at different points in the viral life cycle, is important for normal progression of VZV replication. - Highlights: • Mutation of IE62 domain I did not affect VZV replication in melanoma cells. • IE62 domain II and III are important for VZV replication in melanoma cells. • Mutations of IE62 domain II (DBD) were lethal for virus replication. • Mutations of IE62 NLS and phosphorylation sites inhibited VZV replication. • NLS and S686A/S722A mutations altered localization of IE62 during early and late infection.

  19. Herpes simplex virus type 1 gene UL14: phenotype of a null mutant and identification of the encoded protein.

    Science.gov (United States)

    Cunningham, C; Davison, A J; MacLean, A R; Taus, N S; Baines, J D

    2000-01-01

    Herpes simplex virus type 1 (HSV-1) gene UL14 is located between divergently transcribed genes UL13 and UL15 and overlaps the promoters for both of these genes. UL14 also exhibits a substantial overlap of its coding region with that of UL13. It is one of the few HSV-1 genes for which a phenotype and protein product have not been described. Using mass spectrometric and immunological approaches, we demonstrated that the UL14 protein is a minor component of the virion tegument of 32 kDa which is expressed late in infection. In infected cells, the UL14 protein was detected in the nucleus at discrete sites within electron-dense nuclear bodies and in the cytoplasm initially in a diffuse distribution and then at discrete sites. Some of the UL14 protein was phosphorylated. A mutant with a 4-bp deletion in the central region of UL14 failed to produce the UL14 protein and generated small plaques. The mutant exhibited an extended growth cycle at low multiplicity of infection and appeared to be compromised in efficient transit of virus particles from the infected cell. In mice injected intracranially, the 50% lethal dose of the mutant was reduced more than 30,000-fold. Recovery of the mutant from the latently infected sacral ganglia of mice injected peripherally was significantly less than that of wild-type virus, suggesting a marked defect in the establishment of, or reactivation from, latent infection.

  20. Urban archaeology: new perspectives and possibilities

    Science.gov (United States)

    Leucci, Giovanni; De Giorgi, Lara; Persico, Raffaele

    2017-04-01

    The study of ancient remains is more difficult in urban environments than in an archaeological site, because the ancient town and the modern one superpose to each other and precious testimonies are present just under the current irremovable roads and the buildings. However, modern techniques allows to investigate the past under the present, and allows to retrieve information and possibly create a fruition of the ancient site. IBAM-CNR has been engaged for years in this kind of problems, making use of GPR, ERT and other geophysical techniques [1-3], virtual reality [4] and minimally invasive diagnostics [5] in several towns, in particular in southern Italy and Sicily. The valorization of sites in urban areas require precise projects, founding and clear ideas and agreements about the management of the cultural heritage, because only in this case the work performed will be really exploited and enjoyed by specialists and common people. At the conference, some examples will be shown regarding monuments in the town of Lecce, Italy. References [1] M. Pieraccini, L. Noferini, D. Mecatti, C. Atzeni, R. Persico, F. Soldovieri, Advanced Processing Techniques for Step-frequency Continuous-Wave Penetrating Radar: the Case Study of "Palazzo Vecchio" Walls (Firenze, Italy), Research on Nondestructive Evaluation, vol. 17, pp. 71-83, 2006. [2] Masini N, Persico R., Rizzo E, Calia A, Giannotta M. T., Quarta G., Pagliuca A., "Integrated Techniques for Analysis and Monitoring of Historical Monuments: the case of S.Giovanni al Sepolcro in Brindisi (Southern Italy)." Near Surface Geophysics, vol. 8, n. 5, pp. 423-432, 2010. [3] G. Leucci, N. Masini, R. Persico, F. Soldovieri." GPR and sonic tomography for structural restoration : the case of the Cathedral of Tricarico", Journal of Geophysics and Engineering, vol. 8, pp. S76-S92, Aug. 2011. [4] F. Gabellone, G. Leucci, N. Masini, R. Persico, G. Quarta, F. Grasso, "Nondestructive Prospecting and virtual reconstruction of the chapel of the

  1. Modelflow Estimates of Stroke Volume Do Not Correlate With Doppler Ultrasound Estimates During Upright Posture

    Science.gov (United States)

    Ferguson, Connor R.; Lee, Stuart M. C.; Stenger, Michael B.; Platts, Steven H.; Laurie, Steven S.

    2014-01-01

    Orthostatic intolerance affects 60-80% of astronauts returning from long-duration missions, representing a significant risk to completing mission-critical tasks. While likely multifactorial, a reduction in stroke volume (SV) represents one factor contributing to orthostatic intolerance during stand and head up tilt (HUT) tests. Current measures of SV during stand or HUT tests use Doppler ultrasound and require a trained operator and specialized equipment, restricting its use in the field. BeatScope (Finapres Medical Systems BV, The Netherlands) uses a modelflow algorithm to estimate SV from continuous blood pressure waveforms in supine subjects; however, evidence supporting the use of Modelflow to estimate SV in subjects completing stand or HUT tests remain scarce. Furthermore, because the blood pressure device is held extended at heart level during HUT tests, but allowed to rest at the side during stand tests, changes in the finger arterial pressure waveform resulting from arm positioning could alter modelflow estimated SV. The purpose of this project was to compare Doppler ultrasound and BeatScope estimations of SV to determine if BeatScope can be used during stand or HUT tests. Finger photoplethysmography was used to acquire arterial pressure waveforms corrected for hydrostatic finger-to-heart height using the Finometer (FM) and Portapres (PP) arterial pressure devices in 10 subjects (5 men and 5 women) during a stand test while simultaneous estimates of SV were collected using Doppler ultrasound. Measures were made after 5 minutes of supine rest and while subjects stood for 5 minutes. Next, SV estimates were reacquired while each arm was independently raised to heart level, a position similar to tilt testing. Supine SV estimates were not significantly different between all three devices (FM: 68+/-20, PP: 71+/-21, US: 73+/-21 ml/beat). Upon standing, the change in SV estimated by FM (-18+/-8 ml) was not different from PP (-21+/-12), but both were significantly

  2. Multidisciplinary Investigations embedded in a photogrammatric three dimensional survey in an archaeological site and St Peter and Paul Church in Agro Valley (Messina, Italy)

    Science.gov (United States)

    Crupi, Vincenza; D'Amico, Sebastiano; Majolino, Domenico; Paladini, Giuseppe; Persico, Raffaele; Saccone, Mauro; Spagnolo, Grazia; Venuti, Valentina

    2017-04-01

    for Step-frequency Continuous-Wave Penetrating Radar: the Case Study of "Palazzo Vecchio" Walls (Firenze, Italy), Research on Nondestructive Evaluation, vol. 17, pp. 71-83, 2006 [2] Masini N, Persico R., Rizzo E, Calia A, Giannotta M. T., Quarta G., Pagliuca A., "Integrated Techniques for Analysis and Monitoring of Historical Monuments: the case of S.Giovanni al Sepolcro in Brindisi (Southern Italy)." Near Surface Geophysics, vol. 8 (5), pp. 423-432, 2010. [3] F. Soldovieri, R. Persico and G. Leone, "Effect of source and receiver radiation characteristics in subsurface prospecting within the DBA", Radio Science, vol. 40, RS3006, May 2005.

  3. Efeito de substratos na formação de porta-enxertos de Psidium guajava L. cv. Ogawa em tubetes Effect of substrates in the formation of Psidium guajava L. cv. Ogawa rootstocks in root plugs

    Directory of Open Access Journals (Sweden)

    Diva Correia

    2005-04-01

    Full Text Available Avaliou-se a formação de porta-enxertos de goiabeira em tubetes e em diferentes substratos: vermiculita + vermicomposto, casca de arroz carbonizada + vermicomposto, vermicomposto + tegumento de amêndoa de caju decomposto, casca de arroz carbonizada + esterco bovino curtido, casca de arroz carbonizada + vermiculita e pó da casca de coco maduro + folha de carnaubeira triturada e decomposta. Utilizou-se o delineamento experimental inteiramente casualizado, com 6 tratamentos, 4 repetições e 14 plantas por parcela. Foram avaliados a altura da parte aérea, o diâmetro do caule e o número de folhas aos 60; 90 e 120 dias após a semeadura. Avaliaram-se a massa da matéria seca da parte aérea e das raízes, a facilidade de retirada da muda do tubete e a agregação do substrato às raízes, aos 120 dias. Porta-enxertos produzidos aos 120 dias propiciaram os maiores valores de altura da parte aérea (30,2 cm, diâmetro do caule (5,3 mm, massa da matéria seca da parte aérea (5,7 g e das raízes (3,0 g, e número de folhas (14,5 quando foi usado o substrato vermiculita + vermicomposto. Maior facilidade de retirada da muda do tubete foi encontrada nos substratos vermiculita + vermicomposto, vermicomposto + tegumento de amêndoa de caju decomposto, casca de arroz carbonizada + vermiculita e casca de arroz carbonizada + esterco bovino curtido. Todos os substratos apresentaram ótima agregação do substrato às raízes.The formation of guava rootstocks in root plugs was evaluated in differents substrates: vermiculite + vermicompost, carbonized rice hulls + vermicompost, vermicompost + decompose cashewnut tegument, carbonized rice hulls + cattle manure, carbonized rice hulls + vermiculite and dust peel of ripened coconut fruit + decompose leave dust from carnauba palm tree. The experimental design was completely randomized with 6 treatments, 4 replicates and 14 plants per plot. The plant height, stem diameter and number of leaves were measured in

  4. Identification of herpesvirus proteins that contribute to G1/S arrest.

    Science.gov (United States)

    Paladino, Patrick; Marcon, Edyta; Greenblatt, Jack; Frappier, Lori

    2014-04-01

    Lytic infection by herpesviruses induces cell cycle arrest at the G1/S transition. This appears to be a function of multiple herpesvirus proteins, but only a minority of herpesvirus proteins have been examined for cell cycle effects. To gain a more comprehensive understanding of the viral proteins that contribute to G1/S arrest, we screened a library of over 200 proteins from herpes simplex virus type 1, human cytomegalovirus, and Epstein-Barr virus (EBV) for effects on the G1/S interface, using HeLa fluorescent, ubiquitination-based cell cycle indicator (Fucci) cells in which G1/S can be detected colorimetrically. Proteins from each virus were identified that induce accumulation of G1/S cells, predominantly tegument, early, and capsid proteins. The identification of several capsid proteins in this screen suggests that incoming viral capsids may function to modulate cellular processes. The cell cycle effects of selected EBV proteins were further verified and examined for effects on p53 and p21 as regulators of the G1/S transition. Two EBV replication proteins (BORF2 and BMRF1) were found to induce p53 but not p21, while a previously uncharacterized tegument protein (BGLF2) was found to induce p21 protein levels in a p53-independent manner. Proteomic analyses of BGLF2-interacting proteins identified interactions with the NIMA-related protein kinase (NEK9) and GEM-interacting protein (GMIP). Silencing of either NEK9 or GMIP induced p21 without affecting p53 and abrogated the ability of BGLF2 to further induce p21. Collectively, these results suggest multiple viral proteins contribute to G1/S arrest, including BGLF2, which induces p21 levels likely by interfering with the functions of NEK9 and GMIP. Most people are infected with multiple herpesviruses, whose proteins alter the infected cells in several ways. During lytic infection, the viral proteins block cell proliferation just before the cellular DNA replicates. We used a novel screening method to identify proteins

  5. Viral FGARAT ORF75A promotes early events in lytic infection and gammaherpesvirus pathogenesis in mice

    Science.gov (United States)

    Hogan, Chad H.; Oldenburg, Darby G.; Kara, Mehmet

    2018-01-01

    Gammaherpesviruses encode proteins with homology to the cellular purine metabolic enzyme formyl-glycinamide-phosphoribosyl-amidotransferase (FGARAT), but the role of these viral FGARATs (vFGARATs) in the pathogenesis of a natural host has not been investigated. We report a novel role for the ORF75A vFGARAT of murine gammaherpesvirus 68 (MHV68) in infectious virion production and colonization of mice. MHV68 mutants with premature stop codons in orf75A exhibited a log reduction in acute replication in the lungs after intranasal infection, which preceded a defect in colonization of multiple host reservoirs including the mediastinal lymph nodes, peripheral blood mononuclear cells, and the spleen. Intraperitoneal infection rescued splenic latency, but not reactivation. The 75A.stop virus also exhibited defective replication in primary fibroblast and macrophage cells. Viruses produced in the absence of ORF75A were characterized by an increase in the ratio of particles to PFU. In the next round of infection this led to the alteration of early events in lytic replication including the deposition of the ORF75C tegument protein, the accelerated kinetics of viral gene expression, and induction of TNFα release and cell death. Infecting cells to deliver equivalent genomes revealed that ORF75A was required for initiating early events in infection. In contrast with the numerous phenotypes observed in the absence of ORF75A, ORF75B was dispensable for replication and pathogenesis. These studies reveal that murine rhadinovirus vFGARAT family members ORF75A and ORF75C have evolved to perform divergent functions that promote replication and colonization of the host. PMID:29390024

  6. Morpho-anatomy, imbibition, viability and germination of the seed of Anadenanthera colubrina var. cebil (Fabaceae).

    Science.gov (United States)

    Varela, Rodolfo Omar; Albornoz, Patricia Liliana

    2013-09-01

    Seed biology is a relevant aspect of tropical forests because it is central to the understanding of processes of plant establishment, succession and natural regeneration. Anadenanthera colubrina var. cebil is a timber tree from South America that produces large seeds with thin weak teguments, which is uncommon among legumes. This study describes the morphology and anatomy of the seed coat, the viability, imbibition, and germination in this species. Seeds used during the essays came from 10 trees that grow naturally in Horco Molle, province of Tucumán, Argentina. Seed morphology was described from a sample of 20 units. The seed coat surface was examined with a scanning electron microscope. Transverse sections of hydrated and non-hydrated seeds were employed to describe the histological structure of the seed coat. Hydration, viability and germination experiments were performed under laboratory controlled conditions; and the experimental design consisted of 10 replicas of 10 seeds each. Viability and germination tests were conducted using freshly fallen seeds and seeds stored for five months. Morphologically the seeds of A. colubrina var. cebil are circular to subcircular, laterally compressed, smooth, bright brown and have a horseshoe fissure line (= pleurogram) on both sides. The seed coat comprises five tissue layers and a double (external and internal) cuticle. The outer cuticle (on the epidermis) is smooth and interrupted by microcracks and pores of variable depth. The epidermis consists of macroesclereids with non-lignified secondary walls. This layer is separated from the underlying ones during seed hydration. The other layers of internal tissues are comprised of osteosclereids, parenchyma, osteosclereids, and macrosclereids. The percentage of viable seeds was 93%, decreasing to 75% in seeds with five months old. Seed mass increased 76% after the first eight hours of hydration. Germination percentage was 75% after 76 hours. Germination of seeds stored for five

  7. Relação entre características morfológicas da cariopse e fusariose em milho Relationship between caryopsis morphological characteristics and fusarium ear rot in corn

    Directory of Open Access Journals (Sweden)

    Raquel Silva Costa

    2003-01-01

    Full Text Available Fusarium moniliforme causa sérios prejuízos no rendimento do milho. Suspeita-se que a entrada do fungo na cariopse ocorra pela cicatriz do estilete, e que este fato e o nível de infestação por F. moniliforme estejam relacionados com características morfológicas da cariopse. O objetivo deste trabalho foi identificar características morfológicas que conferem resistência a F. moniliforme em cariopses de milho. Cariopses de seis linhagens de milho, com e sem inoculação do patógeno, foram avaliadas utilizando-se de estereomicroscópio e o microscópio eletrônico de varredura. Características morfológicas da cariopse, tais como: tegumento com saliências e reentrâncias acentuadas, pericarpo pouco espesso, amido menos compacto e presença do canal estilar favorecem a penetração de F. moniliforme.Fusarium ear rot of corn caused by Fusarium moniliforme is responsible for severe yield losses. Probably fungal penetration in the caryopsis takes place at the silk scar and is related to the level of infection and the morphological characteristics of the caryopsis. The objective of this work was to identify morphological characteristics that confer resistance to the fungus in caryopsis of corn. Caryopsis of six lines of corn, with and without inoculation with the pathogen, were evaluated using a stereomicroscope and scanning electron microscope. Morphological characteristics of the caryopsis such as tegument with saliences and reentrances accentuated, pericarp no much thick, starch less compact and presence of the silk canal favor the penetration of F. moniliforme.

  8. SOMATIC EMBRYOGENESIS AND MORPHOANATOMY OF Ocotea porosa SOMATIC EMBRYOS

    Directory of Open Access Journals (Sweden)

    Luciana Luiza Pelegrini

    2013-01-01

    Full Text Available Ocotea porosa seeds have strong tegument dormancy, recalcitrant behavior, low and irregular germination and that makes its natural propagation difficult. The aim of this study was to establish a protocol of regeneration of Ocotea porosa from somatic embryogenesis. Immature embryonic axes were inoculated on WPM culture medium supplemented with 2.4-D (200 μM combined or not with hydrolyzed casein or glutamine (0.5 or 1 g l-1, during 90 days. The repetitive embryogenesis was induced on medium with 2.4-D (22.62 μM combined with 2-iP (2.46 μM followed by transfer to culture medium with hydrolyzed casein or glutamine (1 g l-1 during 90 days. The maturation of somatic embryos was tested in culture medium containing NAA (0.5 μM and 2-iP (5; 10 and 20 μM. The highest percentage of somatic embryos induction (8.3% was observed in WPM culture medium containing 200 μM 2.4-D and 1 g L-1 hydrolyzed casein and the development of somatic embryos occurred indirectly. Repetitive somatic embryogenesis was promoted in WPM medium containing hydrolyzed casein or glutamine. However, the culture medium containing hydrolyzed casein promoted the maintenance of embryogenic capacity for more than two years. During the maturity phase, there was a low progression of globular embryos to cordiform and torpedo stages. The different ontogenetic stages of somatic embryos of Ocotea porosa were characterized by histological studies.

  9. Analysis of complex patterns of human exposure and immunity to Schistosomiasis mansoni: the influence of age, sex, ethnicity and IgE.

    Directory of Open Access Journals (Sweden)

    Angela Pinot de Moira

    2010-09-01

    Full Text Available Numerous factors may influence Schistosoma infection intensity and prevalence within endemic communities, including exposure-related factors such as local environment and behaviour, and factors relating to susceptibility to infection such as immunology and genetics. While animal studies performed in the laboratory can be tightly controlled, human populations are highly heterogeneous, varying according to demographic characteristics, genetic background and exposure to infection. The heterogeneous nature of human water contact behaviour in particular makes it difficult to distinguish between a lack of cercarial exposure and reduced susceptibility to infection as the cause for low levels of infection in the field.In this study we investigate risk factors for Schistosoma mansoni infection in a rural Ugandan fishing community receiving treatment as part of a multi-disciplinary longitudinal reinfection study. More specifically, we examine the influence that age, sex and ethnic background have on susceptibility to reinfection after anti-helminth drug treatment, but use individual estimates of cercarial exposure and multivariable methods in an attempt to remove noise created by environmental and behavioural heterogeneities. We then investigate whether schistosome-specific IgE immune responses could account for any remaining variations in susceptibility to reinfection. Our findings suggest that observed ethnic- and sex-related variations in S. mansoni reinfection were due to variations in cercarial exposure, as opposed to biological differences in susceptibility to infection. Age-related differences in reinfection were not explained by exposure, however, and appeared linked to the balance of IgE and IgG(4 to the tegumental antigen SmTAL1 (formerly Sm22.6, which itself was significantly related to resistance to reinfection.This study highlights the benefit of taking a multidisciplinary approach in complex field settings; it allows the ecology of a

  10. Altered patterns of gene expression underlying the enhanced immunogenicity of radiation-attenuated schistosomes.

    Directory of Open Access Journals (Sweden)

    Gary P Dillon

    2008-05-01

    Full Text Available Schistosome cercariae only elicit high levels of protective immunity against a challenge infection if they are optimally attenuated by exposure to ionising radiation that truncates their migration in the lungs. However, the underlying molecular mechanisms responsible for the altered phenotype of the irradiated parasite that primes for protection have yet to be identified.We have used a custom microarray comprising probes derived from lung-stage parasites to compare patterns of gene expression in schistosomula derived from normal and irradiated cercariae. These were transformed in vitro and cultured for four, seven, and ten days to correspond in development to the priming parasites, before RNA extraction. At these late times after the radiation insult, transcript suppression was the principal feature of the irradiated larvae. Individual gene analysis revealed that only seven were significantly down-regulated in the irradiated versus normal larvae at the three time-points; notably, four of the protein products are present in the tegument or associated with its membranes, perhaps indicating a perturbed function. Grouping of transcripts using Gene Ontology (GO and subsequent Gene Set Enrichment Analysis (GSEA proved more informative in teasing out subtle differences. Deficiencies in signalling pathways involving G-protein-coupled receptors suggest the parasite is less able to sense its environment. Reduction of cytoskeleton transcripts could indicate compromised structure which, coupled with a paucity of neuroreceptor transcripts, may mean the parasite is also unable to respond correctly to external stimuli.The transcriptional differences observed are concordant with the known extended transit of attenuated parasites through skin-draining lymph nodes and the lungs: prolonged priming of the immune system by the parasite, rather than over-expression of novel antigens, could thus explain the efficacy of the irradiated vaccine.

  11. Stimulating Neoblast-Like Cell Proliferation in Juvenile Fasciola hepatica Supports Growth and Progression towards the Adult Phenotype In Vitro.

    Science.gov (United States)

    McCusker, Paul; McVeigh, Paul; Rathinasamy, Vignesh; Toet, Hayley; McCammick, Erin; O'Connor, Anna; Marks, Nikki J; Mousley, Angela; Brennan, Gerard P; Halton, David W; Spithill, Terry W; Maule, Aaron G

    2016-09-01

    Fascioliasis (or fasciolosis) is a socioeconomically important parasitic disease caused by liver flukes of the genus Fasciola. Flukicide resistance has exposed the need for new drugs and/or a vaccine for liver fluke control. A rapidly improving 'molecular toolbox' for liver fluke encompasses quality genomic/transcriptomic datasets and an RNA interference platform that facilitates functional genomics approaches to drug/vaccine target validation. The exploitation of these resources is undermined by the absence of effective culture/maintenance systems that would support in vitro studies on juvenile fluke development/biology. Here we report markedly improved in vitro maintenance methods for Fasciola hepatica that achieved 65% survival of juvenile fluke after 6 months in standard cell culture medium supplemented with 50% chicken serum. We discovered that this long-term maintenance was dependent upon fluke growth, which was supported by increased proliferation of cells resembling the "neoblast" stem cells described in other flatworms. Growth led to dramatic morphological changes in juveniles, including the development of the digestive tract, reproductive organs and the tegument, towards more adult-like forms. The inhibition of DNA synthesis prevented neoblast-like cell proliferation and inhibited growth/development. Supporting our assertion that we have triggered the development of juveniles towards adult-like fluke, mass spectrometric analyses showed that growing fluke have an excretory/secretory protein profile that is distinct from that of newly-excysted juveniles and more closely resembles that of ex vivo immature and adult fluke. Further, in vitro maintained fluke displayed a transition in their movement from the probing behaviour associated with migrating stage worms to a slower wave-like motility seen in adults. Our ability to stimulate neoblast-like cell proliferation and growth in F. hepatica underpins the first simple platform for their long-term in vitro study

  12. Immunocytochemical demonstration of neuropeptides in the fish-gill parasite, Diclidophora merlangi (Monogenoidea).

    Science.gov (United States)

    Maule, A G; Halton, D W; Johnston, C F; Fairweather, I; Shaw, C

    1989-05-01

    Using the indirect immunofluorescence technique, immunoreactivity (IR) to three mammalian and one invertebrate regulatory peptide has been demonstrated in the nervous system of the monogenean gill parasite Diclidophora merlangi. IR to pancreatic polypeptide (PP), peptide tyrosine tyrosine (PYY) and FMRFamide was evident throughout central and peripheral nervous tissues, whereas vasoactive intestinal polypeptide (VIP)-IR was confined to a portion of the longitudinal ventral nerve cords. Staining patterns revealed the orthogonal arrangement of the nervous system consisting of paired cerebral ganglia, connecting post-pharyngeal commissure, three pairs of longitudinal nerve cords and associated neurones. PP-IR, PYY-IR and FMRFamide-IR were intense throughout the central nervous system of the worm. A small plexus of nerve fibres and somata in each peduncle was immunoreactive for FMRFamide and provided innervation to each of the eight posterior clamps. In the peripheral nervous system, PP-IR, PYY-IR and FMRFamide-IR occurred in an extensive nerve-net with fine, possibly sensory nerve endings in the tegument. PP-IR was also present in nerve fibres in the walls of the ootype, seminal vesicle and uterus. PYY- and FMRFamide-IRs, while evident in nerve fibres of the ootype wall, were also present in a distinct population of cells that encircles the ootype, and which are linked to it by fine cytoplasmic connectives. The majority of these somata were bipolar or multipolar. PYY-IR and FMRFamide-IR were also associated with nerve fibres and bipolar cells in the wall of the vitelline reservoir. Regulatory peptides would appear to play an integral role in neuronal functioning and egg development in D. merlangi.

  13. Reduction of microbial contamination and improvement of germination of sweet basil (Ocimum basilicum L.) seeds via surface dielectric barrier discharge

    Science.gov (United States)

    Ambrico, Paolo F.; Šimek, Milan; Morano, Massimo; De Miccolis Angelini, Rita M.; Minafra, Angelantonio; Trotti, Pasquale; Ambrico, Marianna; Prukner, Václav; Faretra, Francesco

    2017-08-01

    Naturally contaminated basil seeds were treated by a surface dielectric barrier discharge driven in the humid air by an amplitude modulated AC high voltage to avoid heat shock. In order to avoid direct contact of seeds with microdischarge filaments, the seeds to be treated were placed at sufficient distance from the surface discharge. After treatment, the seeds were analyzed in comparison with control samples for their microbial contamination as well as for the capability of germination and seedling growth. Moreover, chemical modification of seed surface was observed through the elemental energy dispersive x-ray analysis and wettability tests. We found that treatment applied at 20% duty cycle (effective discharge duration up to 20 s) significantly decreases microbial load without reducing the viability of the seeds. On the other side, seedling growth was considerably accelerated after the treatment, and biometric growth parameters of seedlings (total length, weight, leaf extension) considerably increased compared to the controls. Interestingly, scanning electron microscopy images taken for the different duration of treatment revealed that seed radicle micropylar regions underwent significant morphological changes while the coat was substantially undamaged. Inside the seed, the embryo seemed to be well preserved while the endosperm body was detached from the epithelial tegument. A total of 9 different genera of fungi were recovered from the analyzed seeds. Scanning electron microscopy images revealed that conidia were localized especially in the micropylar region, and after plasma treatment, most of them showed substantial damages. Therefore, the overall effect of the treatment of naturally contaminated seeds by reactive oxygen and nitrogen species produced by plasma and the consequent changes in surface chemistry and microbial load can significantly improve seed vigor.

  14. Potentilla fulgens (Family Rosaceae), a medicinal plant of north-east India: a natural anthelmintic?

    Science.gov (United States)

    Roy, Bishnupada; Swargiary, Ananta; Syiem, D; Tandon, V

    2010-10-01

    The cestode parasite, Raillietina echinobothrida and the trematode, Gastrothylax crumenifer were exposed to the ethanolic root peel extract of Potentilla fulgens, an antiparasitic local medicinal plant of Meghalaya, India, to evaluate the anthelmintic efficacy of the plant. The parasites were incubated in 1, 5, 10, 20, 50 and 100 mg crude alcoholic extract per ml of phosphate buffered saline (PBS) at a temperature of 37 ± 1°C. Paralysis and death were observed at 2.00 ± 0.05 and 2.80 ± 0.06 h for the cestode and 1.21 ± 0.06 and 2.18 ± 0.04 h for the trematode parasites at the highest test concentration of the plant extract. The commercial anthelmintic, Praziquantel (PZQ) showed higher activity at the tested concentration (0.02 mg/ml). To further investigate the efficacy of the plant extract, vital tegumental enzymes of the parasite viz. Acid phosphatase (AcPase), Alkaline phosphatase (AlkPase) and Adenosine triphosphatase (ATPase) were studied. Quantitatively, the total enzyme activity of AcPase, AlkPase and ATPase was found to be reduced significantly by 69.20, 66.43 and 29.63% for R. echinobothrida and 47.96, 51.79 and 42.63% for G. crumenifer, respectively compared to the respective controls; histochemical study also showed reduction in the visible staining of the enzymes. The reference drug, PZQ also showed more or less similar effect like that of the plant extract. The result suggests that phytochemicals of P. fulgens have anthelmintic potential.

  15. Anthelmintic Potential of Thymoquinone and Curcumin on Fasciola gigantica.

    Directory of Open Access Journals (Sweden)

    Rizwan Ullah

    Full Text Available Fasciolosis an economically important global disease of ruminants in the temperate and tropical regions, caused by Fasciola hepatica and F. gigantica, respectively, also poses a potential zoonotic threat. In India alone it causes huge losses to stakeholders. Anthelmintics including triclabendazole have been used to control this menace but the emerging resistance against the available compounds necessitates identification of novel and alternative therapeutic measures involving plant derived natural compounds for their anthelmintic potential. Thymoquinone (T and curcumin (C, the active ingredients of Nigella sativa and Curcuma longa respectively have been used as antiparasitic agents but the information on their flukicidal effect is very limited. Adult flukes of F. gigantica were in vitro exposed to different concentrations of thymoquinone and curcumin separately for 3h at 37+ 1°C. A significant (p<0.05 reduction in the worm motility at 60 μM concentration of both T and C was observed though all the worms remained alive after 3h exposure, whereas the effect on egg shedding was statistically insignificant. Pronounced tegumental disruptions and erosion of spines in the posterior region and around the acetabulum was evident. A significant (p<0.05 decrease in glutathione-S-transferase and superoxide dismutase activity and reduced glutathione (GSH level was observed, while protein carbonylation increased differentially. A significant inhibition of CathepsinL (CatL gene expression in thymoquinone treated worms was also evident. Further, in silico molecular docking of T and C with CatL revealed a stronger interaction of curcumin with the involvement of higher number of amino acids as compared to thymoquinone that could be more effective in inhibiting the antioxidant enzymes of F. gigantica. It is concluded that both the compounds understudy will decrease the detoxification ability of F. gigantica, while inhibition of CatL will significantly affect

  16. De Novo Herpes Simplex Virus VP16 Expression Gates a Dynamic Programmatic Transition and Sets the Latent/Lytic Balance during Acute Infection in Trigeminal Ganglia.

    Science.gov (United States)

    Sawtell, Nancy M; Thompson, Richard L

    2016-09-01

    The life long relationship between herpes simplex virus and its host hinges on the ability of the virus to aggressively replicate in epithelial cells at the site of infection and transport into the nervous system through axons innervating the infection site. Interaction between the virus and the sensory neuron represents a pivot point where largely unknown mechanisms lead to a latent or a lytic infection in the neuron. Regulation at this pivot point is critical for balancing two objectives, efficient widespread seeding of the nervous system and host survival. By combining genetic and in vivo in approaches, our studies reveal that the balance between latent and lytic programs is a process occurring early in the trigeminal ganglion. Unexpectedly, activation of the latent program precedes entry into the lytic program by 12 -14hrs. Importantly, at the individual neuronal level, the lytic program begins as a transition out of this acute stage latent program and this escape from the default latent program is regulated by de novo VP16 expression. Our findings support a model in which regulated de novo VP16 expression in the neuron mediates entry into the lytic cycle during the earliest stages of virus infection in vivo. These findings support the hypothesis that the loose association of VP16 with the viral tegument combined with sensory axon length and transport mechanisms serve to limit arrival of virion associated VP16 into neuronal nuclei favoring latency. Further, our findings point to specialized features of the VP16 promoter that control the de novo expression of VP16 in neurons and this regulation is a key component in setting the balance between lytic and latent infections in the nervous system.

  17. Acanthocephalus amini n. sp. (Acanthocephala: Echinorhynchidae) from the freshwater fish Cichlasoma urophthalmus (Günther) (Cichlidae) in Mexico.

    Science.gov (United States)

    Salgado-Maldonado, Guillermo; Novelo-Turcotte, María Teresa

    2009-07-01

    Acanthocephalus amini n. sp. (Palaeacanthocephala: Echinorhynchidae) is described from the intestine of Cichlasoma urophthalmus (Günther) (Pisces: Cichlidae) collected in the Río Champotón, a river in Campeche State, Mexico. It is the fourth species of Acanthocephalus Koelreuther, 1771 described from North American freshwater fishes, although two other species are known from South America. The new species is distinguished from other members of Acanthocephalus by features of its trunk, which is small, clavate, slightly expanded medially and bluntly pointed posteriorly. It is further distinguished by having a cylindrical proboscis armed with 13-14 longitudinal rows of 11-12 stout hooks; the apical and medial proboscis hooks are almost uniform in size and shape, decreasing in size towards the base; the posteriormost hooks are smaller, straighter and more slender than the anterior and middle hooks; and the lateral rows of hooks are more widely spaced, forming a conspicuous longitudinal area devoid of hooks. Furthermore, the lemnisci are saccate and shorter than the proboscis receptacle; and the neck is very short with a thick collar of trunk tegument, which encircles the base of proboscis. In males, the testes are in the middle third of trunk, diagonal, spherical and small relative to the body size, and there are six clavate cement glands. In females, the uterus forms a conspicuous, elongate, cylindrical egg reservoir. The new species is most similar to A. alabamensis Amin & Williams, 1983, but can be distinguished by its swollen, clavate trunk, the largest proboscis hooks being present apically and medially, smaller testes, a shorter male reproductive system relative to body size and females with a prominent uterus. They have different hosts and geographical distribution. The new species can be differentiated from Brasacanthus sphoeroides Thatcher, 2001, a similar species in a monotypic echinorhynchid genus, because the latter is larger, has smaller proboscis hooks

  18. Herpesvirus gB-induced fusion between the virion envelope and outer nuclear membrane during virus egress is regulated by the viral US3 kinase.

    Science.gov (United States)

    Wisner, Todd W; Wright, Catherine C; Kato, Akihisa; Kawaguchi, Yasushi; Mou, Fan; Baines, Joel D; Roller, Richard J; Johnson, David C

    2009-04-01

    Herpesvirus capsids collect along the inner surface of the nuclear envelope and bud into the perinuclear space. Enveloped virions then fuse with the outer nuclear membrane (NM). We previously showed that herpes simplex virus (HSV) glycoproteins gB and gH act in a redundant fashion to promote fusion between the virion envelope and the outer NM. HSV mutants lacking both gB and gH accumulate enveloped virions in herniations, vesicles that bulge into the nucleoplasm. Earlier studies had shown that HSV mutants lacking the viral serine/threonine kinase US3 also accumulate herniations. Here, we demonstrate that HSV gB is phosphorylated in a US3-dependent manner in HSV-infected cells, especially in a crude nuclear fraction. Moreover, US3 directly phosphorylated the gB cytoplasmic (CT) domain in in vitro assays. Deletion of gB in the context of a US3-null virus did not add substantially to defects in nuclear egress. The majority of the US3-dependent phosphorylation of gB involved the CT domain and amino acid T887, a residue present in a motif similar to that recognized by US3 in other proteins. HSV recombinants lacking gH and expressing either gB substitution mutation T887A or a gB truncated at residue 886 displayed substantial defects in nuclear egress. We concluded that phosphorylation of the gB CT domain is important for gB-mediated fusion with the outer NM. This suggested a model in which the US3 kinase is incorporated into the tegument layer (between the capsid and envelope) in HSV virions present in the perinuclear space. By this packaging, US3 might be brought close to the gB CT tail, leading to phosphorylation and triggering fusion between the virion envelope and the outer NM.

  19. Reduction of microbial contamination and improvement of germination of sweet basil ( Ocimum basilicum L.) seeds via surface dielectric barrier discharge

    International Nuclear Information System (INIS)

    Ambrico, Paolo F; Ambrico, Marianna; Šimek, Milan; Prukner, Václav; Morano, Massimo; De Miccolis Angelini, Rita M; Trotti, Pasquale; Faretra, Francesco; Minafra, Angelantonio

    2017-01-01

    Naturally contaminated basil seeds were treated by a surface dielectric barrier discharge driven in the humid air by an amplitude modulated AC high voltage to avoid heat shock. In order to avoid direct contact of seeds with microdischarge filaments, the seeds to be treated were placed at sufficient distance from the surface discharge. After treatment, the seeds were analyzed in comparison with control samples for their microbial contamination as well as for the capability of germination and seedling growth. Moreover, chemical modification of seed surface was observed through the elemental energy dispersive x-ray analysis and wettability tests. We found that treatment applied at 20% duty cycle (effective discharge duration up to 20 s) significantly decreases microbial load without reducing the viability of the seeds. On the other side, seedling growth was considerably accelerated after the treatment, and biometric growth parameters of seedlings (total length, weight, leaf extension) considerably increased compared to the controls. Interestingly, scanning electron microscopy images taken for the different duration of treatment revealed that seed radicle micropylar regions underwent significant morphological changes while the coat was substantially undamaged. Inside the seed, the embryo seemed to be well preserved while the endosperm body was detached from the epithelial tegument. A total of 9 different genera of fungi were recovered from the analyzed seeds. Scanning electron microscopy images revealed that conidia were localized especially in the micropylar region, and after plasma treatment, most of them showed substantial damages. Therefore, the overall effect of the treatment of naturally contaminated seeds by reactive oxygen and nitrogen species produced by plasma and the consequent changes in surface chemistry and microbial load can significantly improve seed vigor. (paper)

  20. PHYSIOLOGICAL QUALITY OF Araucaria angustifolia (Bertol. Kuntze SEEDS SUBMITTED TO DIFFERENT STORAGE CONDITIONS AND ESCARIFICATION

    Directory of Open Access Journals (Sweden)

    Álvaro Valente Caçola

    2010-08-01

    Full Text Available The seeds of Araucaria angustifolia (Bertol. Kuntze are recalcitrants and, therefore, they lose rapidly the viability after the harvest, limiting their use for nursery production of seedlings. This work was carried out to investigate the effects of cold storage duration, conservation method, and escarification on germination and vigour of seeds of Araucaria angustifolia. The seeds were cold stored (0-1oC/90-95% RH for 0, 60, 120, and 180 days, under normal air condictions (NA, modified atmosphere (MA, and controlled atmosphere (CA storage. For each cold storage duration, after removal from cold storage, the seeds were submitted or not to scarification (by cutting a small portion of the apical seed tegument, avoiding the damage of the endosperm, letting the germinate in a chamber with controlled environment, in plastic trays filled with vermiculite, for 60 days. The experiment followed the completely randomized factorial design (4x3x2, with four cold storage durations (0, 60, 120, and 180 days, three storage methods (NA, MA, and CA, with or without escarification, and four replicates. There was a good preservation of physiological quality for seeds submitted to different conservation methods in cold storage along the 180-day period. However, there was a faster germination and initial growth of seedlings for seeds left in cold storage for 60 days than in seeds assessed at harvest or left in cold storage for 120-180 days. The NA storage of seeds in perfurated plastic bags was as efficient as MA and CA storage to preserve seeds quality. Seeds scarification increased vigor and promoted seedlings initial growth of Araucaria angustifolia after being removed from cold storage.

  1. Anthelmintic activity in vivo of epiisopiloturine against juvenile and adult worms of Schistosoma mansoni.

    Directory of Open Access Journals (Sweden)

    Maria A Guimarães

    2015-03-01

    Full Text Available Schistosomiasis is a serious disease currently estimated to affect more that 207 million people worldwide. Due to the intensive use of praziquantel, there is increasing concern about the development of drug-resistant strains. Therefore, it is necessary to search for and investigate new potential schistosomicidal compounds. This work reports the in vivo effect of the alkaloid epiisopiloturine (EPI against adults and juvenile worms of Schistosoma mansoni. EPI was first purified its thermal behavior and theoretical solubility parameters charaterised. In the experiment, mice were treated with EPI over the 21 days post-infection with the doses of 40 and 200 mg/kg, and 45 days post-infection with single doses of 40, 100 and 300 mg/kg. The treatment with EPI at 40 mg/kg was more effective in adult worms when compared with doses of 100 and 300 mg/kg. The treatment with 40 mg/kg in adult worms reduced parasite burden significantly, lead to reduction in hepatosplenomegaly, reduced the egg burden in faeces, and decreased granuloma diameter. Scanning electron microscopy revealed morphological changes to the parasite tegument after treatment, including the loss of important features. Additionally, the in vivo treatment against juvenile with 40 mg/kg showed a reduction of the total worm burden of 50.2%. Histopathological studies were performed on liver, spleen, lung, kidney and brain and EPI was shown to have a DL50 of 8000 mg/kg. Therefore EPI shows potential to be used in schistosomiasis treatment. This is the first time that schistosomicidal in vivo activity of EPI has been reported.

  2. Androgens Exert a Cysticidal Effect upon Taenia crassiceps by Disrupting Flame Cell Morphology and Function

    Science.gov (United States)

    Ambrosio, Javier R.; Valverde-Islas, Laura; Nava-Castro, Karen E.; Palacios- Arreola, M. Isabel; Ostoa-Saloma, Pedro; Reynoso-Ducoing, Olivia; Escobedo, Galileo; Ruíz-Rosado, Azucena; Dominguez-Ramírez, Lenin; Morales-Montor, Jorge

    2015-01-01

    The effects of testosterone (T4) and dihydrotestosterone (DHT) on the survival of the helminth cestode parasite Taenia crassiceps, as well as their effects on actin, tubulin and myosin expression and their assembly into the excretory system of flame cells are described in this paper. In vitro evaluations on parasite viability, flow cytometry, confocal microscopy, video-microscopy of live flame cells, and docking experiments of androgens interacting with actin, tubulin, and myosin were conducted. Our results show that T4 and DHT reduce T. crassiceps viability in a dose- and time-dependent fashion, reaching 90% of mortality at the highest dose used (40 ng/ml) and time exposed (10 days) in culture. Androgen treatment does not induce differences in the specific expression pattern of actin, tubulin, and myosin isoforms as compared with control parasites. Confocal microscopy demonstrated a strong disruption of the parasite tegument, with reduced assembly, shape, and motion of flame cells. Docking experiments show that androgens are capable of affecting parasite survival and flame cell morphology by directly interacting with actin, tubulin and myosin without altering their protein expression pattern. We show that both T4 and DHT are able to bind actin, tubulin, and myosin affecting their assembly and causing parasite intoxication due to impairment of flame cell function. Live flame cell video microscopy showing a reduced motion as well changes in the shape of flame cells are also shown. In summary, T4 and DHT directly act on T. crassiceps cysticerci through altering parasite survival as well as the assembly and function of flame cells. PMID:26076446

  3. Cu,Zn superoxide dismutase: cloning and analysis of the Taenia solium gene and Taenia crassiceps cDNA.

    Science.gov (United States)

    Parra-Unda, Ricardo; Vaca-Paniagua, Felipe; Jiménez, Lucia; Landa, Abraham

    2012-01-01

    Cytosolic Cu,Zn superoxide dismutase (Cu,Zn-SOD) catalyzes the dismutation of superoxide (O(2)(-)) to oxygen and hydrogen peroxide (H(2)O(2)) and plays an important role in the establishment and survival of helminthes in their hosts. In this work, we describe the Taenia solium Cu,Zn-SOD gene (TsCu,Zn-SOD) and a Taenia crassiceps (TcCu,Zn-SOD) cDNA. TsCu,Zn-SOD gene that spans 2.841 kb, and has three exons and two introns; the splicing junctions follow the GT-AG rule. Analysis in silico of the gene revealed that the 5'-flanking region has three putative TATA and CCAAT boxes, and transcription factor binding sites for NF1 and AP1. The transcription start site was a C, located at 22 nucleotides upstream of the translation start codon (ATG). Southern blot analysis showed that TcCu,Zn-SOD and TsCu,Zn-SOD genes are encoded by a single copy. The deduced amino acid sequences of TsCu,Zn-SOD gene and TcCu,Zn-SOD cDNA reveal 98.47% of identity, and the characteristic motives, including the catalytic site and β-barrel structure of the Cu,Zn-SOD. Proteomic and immunohistochemical analysis indicated that Cu,Zn-SOD does not have isoforms, is distributed throughout the bladder wall and is concentrated in the tegument of T. solium and T. crassiceps cysticerci. Expression analysis revealed that TcCu,Zn-SOD mRNA and protein expression levels do not change in cysticerci, even upon exposure to O(2)(-) (0-3.8 nmol/min) and H(2)O(2) (0-2mM), suggesting that this gene is constitutively expressed in these parasites. Published by Elsevier Inc.

  4. In Vitro Study of Taenia solium Postoncospheral Form.

    Directory of Open Access Journals (Sweden)

    Nancy Chile

    2016-02-01

    Full Text Available The transitional period between the oncosphere and the cysticercus of Taenia solium is the postoncospheral (PO form, which has not yet been completely characterized. The aim of this work was to standardize a method to obtain T. solium PO forms by in vitro cultivation. We studied the morphology of the PO form and compared the expression of antigenic proteins among the PO form, oncosphere, and cysticerci stages.T. solium activated oncospheres were co-cultured with ten cell lines to obtain PO forms, which we studied at three stages of development--days 15, 30, and 60. A high percentage (32% of PO forms was obtained using HCT-8 cells in comparison to the other cell lines. The morphology was observed by bright field, scanning, and transmission electron microscopy. Morphology of the PO form changed over time, with the six hooks commonly seen in the oncosphere stage disappearing in the PO forms, and vesicles and microtriches observed in the tegument. The PO forms grew as they aged, reaching a diameter of 2.5 mm at 60 days of culture. 15-30 day PO forms developed into mature cysticerci when inoculated into rats. Antigenic proteins expressed in the PO forms are also expressed by the oncosphere and cysticerci stages, with more cysticerci antigenic proteins expressed as the PO forms ages.This is the first report of an in vitro production method of T. solium PO forms. The changes observed in protein expression may be useful in identifying new targets for vaccine development. In vitro culture of PO form will aid in understanding the host-parasite relationship, since the structural changes of the developing PO forms may reflect the parasite's immunoprotective mechanisms. A wider application of this method could significantly reduce the use of animals, and thus the costs and time required for further experimental investigations.

  5. In Vitro Study of Taenia solium Postoncospheral Form.

    Science.gov (United States)

    Chile, Nancy; Clark, Taryn; Arana, Yanina; Ortega, Ynes R; Palma, Sandra; Mejia, Alan; Angulo, Noelia; Kosek, Jon C; Kosek, Margaret; Gomez-Puerta, Luis A; Garcia, Hector H; Gavidia, Cesar M; Gilman, Robert H; Verastegui, Manuela

    2016-02-01

    The transitional period between the oncosphere and the cysticercus of Taenia solium is the postoncospheral (PO) form, which has not yet been completely characterized. The aim of this work was to standardize a method to obtain T. solium PO forms by in vitro cultivation. We studied the morphology of the PO form and compared the expression of antigenic proteins among the PO form, oncosphere, and cysticerci stages. T. solium activated oncospheres were co-cultured with ten cell lines to obtain PO forms, which we studied at three stages of development--days 15, 30, and 60. A high percentage (32%) of PO forms was obtained using HCT-8 cells in comparison to the other cell lines. The morphology was observed by bright field, scanning, and transmission electron microscopy. Morphology of the PO form changed over time, with the six hooks commonly seen in the oncosphere stage disappearing in the PO forms, and vesicles and microtriches observed in the tegument. The PO forms grew as they aged, reaching a diameter of 2.5 mm at 60 days of culture. 15-30 day PO forms developed into mature cysticerci when inoculated into rats. Antigenic proteins expressed in the PO forms are also expressed by the oncosphere and cysticerci stages, with more cysticerci antigenic proteins expressed as the PO forms ages. This is the first report of an in vitro production method of T. solium PO forms. The changes observed in protein expression may be useful in identifying new targets for vaccine development. In vitro culture of PO form will aid in understanding the host-parasite relationship, since the structural changes of the developing PO forms may reflect the parasite's immunoprotective mechanisms. A wider application of this method could significantly reduce the use of animals, and thus the costs and time required for further experimental investigations.

  6. Endoplasmic reticulum-to-Golgi transitions upon herpes virus infection [version 2; referees: 1 approved, 3 approved with reservations

    Directory of Open Access Journals (Sweden)

    Peter Wild

    2018-02-01

    Full Text Available Background: Herpesvirus capsids are assembled in the nucleus, translocated to the perinuclear space by budding, acquiring tegument and envelope, or released to the cytoplasm via impaired nuclear envelope. One model proposes that envelopment, “de-envelopment” and “re-envelopment” is essential for production of infectious virus. Glycoproteins gB/gH were reported to be essential for de-envelopment, by fusion of the “primary” envelope with the outer nuclear membrane. Yet, a high proportion of enveloped virions generated from genomes with deleted gB/gH were found in the cytoplasm and extracellular space, suggesting the existence of alternative exit routes. Methods: We investigated the relatedness between the nuclear envelope and membranes of the endoplasmic reticulum and Golgi complex, in cells infected with either herpes simplex virus 1 (HSV-1 or a Us3 deletion mutant thereof, or with bovine herpesvirus 1 (BoHV-1 by transmission and scanning electron microscopy, employing freezing technique protocols. Results:  The Golgi complex is a compact entity in a juxtanuclear position covered by a membrane on the cis face. Golgi membranes merge with membranes of the endoplasmic reticulum forming an entity with the perinuclear space. All compartments contained enveloped virions. After treatment with brefeldin A, HSV-1 virions aggregated in the perinuclear space and endoplasmic reticulum, while infectious progeny virus was still produced. Conclusions: The data suggest that virions derived by budding at nuclear membranes are intraluminally transported from the perinuclear space via Golgi -endoplasmic reticulum transitions into Golgi cisternae for packaging. Virions derived by budding at nuclear membranes are infective like Us3 deletion mutants, which  accumulate in the perinuclear space. Therefore, i de-envelopment followed by re-envelopment is not essential for production of infective progeny virus, ii the process taking place at the outer nuclear

  7. Antischistosomal activity of ginger (Zingiber officinale) against Schistosoma mansoni harbored in C57 mice.

    Science.gov (United States)

    Mostafa, Osama M S; Eid, Refaat A; Adly, Mohamed A

    2011-08-01

    The repeated chemotherapy of schistosomiasis has resulted in the emergence of drug-resistant schistosome strains. The development of such resistance has drawn the attention of many authors to alternative drugs. Many medicinal plants were studied to investigate their antischistosomal potency. The present work aimed to evaluate antischistosomal activity of crude aqueous extract of ginger against Schistosoma mansoni. Sixteen mice of C57 strain were exposed to 100 ± 10 cercariae per mouse by the tail immersion method; the mice were divided into two groups: untreated group and ginger-treated one. All mice were sacrificed at the end of 10th week post-infection. Worm recovery and egg counting in the hepatic tissues and faeces were determined. Surface topography of the recovered worms was studied by scanning electron microscopy. Histopathological examination of liver and intestine was done using routine histological procedures. The worm burden and the egg density in liver and faeces of mice treated with ginger were fewer than in non-treated ones. Scanning electron microscopical examination revealed that male worms recovered from mice treated with ginger lost their normal surface architecture, since its surface showed partial loss of tubercles' spines, extensive erosion in inter-tubercle tegumental regions and numerous small blebs around tubercles. Histopathological data indicated a reduction in the number and size of granulomatous inflammatory infiltrations in the liver and intestine of treated mice compared to non-treated mice. The results of the present work suggested that ginger has antischistosomal activities and provided a basis for subsequent experimental and clinical trials.

  8. Polymeric biocomposites obtained from the amylacea and lignocellulose fraction of the mango core (Mangifera indica), Tommy Atkins

    International Nuclear Information System (INIS)

    Cordeiro, Edna Maria Silva

    2013-01-01

    This employment has the function the utilization of mango seeds Tommy Atkins, like starch source to obtain biopolymers and fibers source and nanowhiskers cellulose also, that will be use like reinforcing fillers in micro and nano biocomposites polymeric. The fibers in natura removed from tegument mango seed were characterized, as well as the treated fibers and nanowhiskers of cellulose extracted from them. The starch extract showed a good performance (32%) and a high purity. The chemicals analyzes, of crystallinity and morphological of the fibers in natura, treated fibers and nanowhiskers of cellulose confirmed the efficacy of the chemical treatment performed to remove amorphous constituents (hemicellulose and lignina). The thermoplastic starch (TPS) obtained from two sources, corn starch and starchy material mango, was produced in a twin screw extruder with composition mass of 62,5% of starch, 9,4% of water and 28,1% of glycerol. The starch material mango was the main objective of this work for the production of biodegradable materials, and the starch corn was utilized during the production stage to evaluate the processability of the starch and use as parameter for comparison, according of being a conventional source for obtaining conventional commercial starch. The incorporation of fibers (6% in mass) and nanowhiskers cellulose (1% in mass) in matrix of TPS to obtain composite and nanocomposite, respectively, it was performed in single screw extruder. The biocomposites and bionanocomposites polymeric were obtained and the TPS from starchy material mango presented better results of thermal and mechanicals properties when compared to TPS corn starch. Concludes that the sediment generated of the agroindustrial processing mango used presents potential to producing of biodegradable materials. (author)

  9. In vitro effect of sodium arsenite on Echinococcus granulosus protoscoleces.

    Science.gov (United States)

    Xing, Guoqiang; Wang, Bo; Lei, Ying; Liu, Chunli; Wang, Zhuo; Shi, Hongjuan; Yang, Rentan; Qin, Wenjuan; Jiang, Yufeng; Lv, Hailong

    2016-06-01

    Cystic echinococcosis (CE) caused by the metacestodes of Echinococcus granulosus is an important cosmopolitan zoonosis. Surgery is the main treatment option for CE. Meanwhile, chemotherapy is used as an significant adjunct to surgery. However, the benzimidazole carbamate group and the existing scolicidal agents may not be as effective as hoped. In this study, we aimed to explore the in vitro effect of sodium arsenite (NaAsO2) on Echinococcus granulosus protoscoleces, the causative agents of CE. Protoscoleces of E. granulosus were incubated in vitro with 4, 8, 12, 16, and 20μM NaAsO2. Viability and changes in morphology were investigated by 0.1% eosin staining. The ultrastructural alterations were observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Additionally, caspase-3 activity was measured by colorimetric assay. Obvious protoscolicidal effect was seen with NaAsO2 at concentrations of 16μM and 20μM. Protoscolex mortality was 83.24% (16μM) and 100% (20μM) after 6 days post-incubation. SEM showed that the primary site of drug damage was the tegument of the protoscoleces. TEM analysis demonstrated that the internal tissues were severely affected and revealed an increase in the number of lipid droplets and vacuoles after treatment with 16μM NaAsO2. Meanwhile, the caspase-3 activity significantly increased in protoscoleces after 24h of NaAsO2 incubation compared to the untreated controls. Our study demonstrated the clear in vitro scolicidal effect of NaAsO2 against E. granulosus protoscoleces. However, the in vivo efficacy, specific mechanism, and any possible side effects of NaAsO2 remain to be investigated. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Tratamentos pré-germinativos em sementes de Didymopanax morototoni (Aubl. Dcne. et Planch

    Directory of Open Access Journals (Sweden)

    Elci Terezinha Franco Henz

    2002-01-01

    Full Text Available Didymopanax morototoni (Aubl. Dcne. et Planch, (caixeta, is a native forest tree with economical importance in Brazil. It has a large natural distribution, with rapid growth and is a pioneer species. But the seeds of that species have a low rate of germination. The aim of this work was to evaluate the effect of the pre-treatments on the germination capacity. The following treatments were tested: immersion of seed in water for 8 h; chemical scarification with HCl and H2SO4 a 70%; washing with water and alcohol + water (1:1 at different times (15, 30 and 45 min. and mechanical scarification. In vitro germination tests were also carried out. Seeds without test were inoculated in the WPM medium solidified with agar (6 g/l in presence of kinetin and gibberelic acid on the concentration of 0; 0,1; 0,5; 1,0 mg/l, plus hydrolysed casein on the concentration of 250 mg/l. The incubation was at de 25 ±1°C, in germination chambers under photoperiod of 16 hours light under14, 3 μE.m-². s-¹ or in the darkness. The germination was improved by the pre-treatments rinsed in water or water plus alcohol in 45 minutes. The addition of kinetin and gibberelic acid allowed an improvement in the germination of 40 to 70%. The results confirmed that there was a partial dormancy due to teguments and point out to possible presence of inhibitory substances in them which can be removed by washing the seeds.

  11. Polymeric biocomposites obtained from the amylacea and lignocellulose fraction of the mango core (Mangifera indica), Tommy Atkins; Biocompositos polimericos obtidos a partir da fracao lignocelulosica e amilacea do caroco de manga (Mangifera indica), Tommy Atkins

    Energy Technology Data Exchange (ETDEWEB)

    Cordeiro, Edna Maria Silva

    2013-07-01

    This employment has the function the utilization of mango seeds Tommy Atkins, like starch source to obtain biopolymers and fibers source and nanowhiskers cellulose also, that will be use like reinforcing fillers in micro and nano biocomposites polymeric. The fibers in natura removed from tegument mango seed were characterized, as well as the treated fibers and nanowhiskers of cellulose extracted from them. The starch extract showed a good performance (32%) and a high purity. The chemicals analyzes, of crystallinity and morphological of the fibers in natura, treated fibers and nanowhiskers of cellulose confirmed the efficacy of the chemical treatment performed to remove amorphous constituents (hemicellulose and lignina). The thermoplastic starch (TPS) obtained from two sources, corn starch and starchy material mango, was produced in a twin screw extruder with composition mass of 62,5% of starch, 9,4% of water and 28,1% of glycerol. The starch material mango was the main objective of this work for the production of biodegradable materials, and the starch corn was utilized during the production stage to evaluate the processability of the starch and use as parameter for comparison, according of being a conventional source for obtaining conventional commercial starch. The incorporation of fibers (6% in mass) and nanowhiskers cellulose (1% in mass) in matrix of TPS to obtain composite and nanocomposite, respectively, it was performed in single screw extruder. The biocomposites and bionanocomposites polymeric were obtained and the TPS from starchy material mango presented better results of thermal and mechanicals properties when compared to TPS corn starch. Concludes that the sediment generated of the agroindustrial processing mango used presents potential to producing of biodegradable materials. (author)

  12. Anthelmintic Potential of Thymoquinone and Curcumin on Fasciola gigantica.

    Science.gov (United States)

    Ullah, Rizwan; Rehman, Abdur; Zafeer, Mohd Faraz; Rehman, Lubna; Khan, Yasir A; Khan, M A Hannan; Khan, Shahper N; Khan, Asad U; Abidi, S M A

    2017-01-01

    Fasciolosis an economically important global disease of ruminants in the temperate and tropical regions, caused by Fasciola hepatica and F. gigantica, respectively, also poses a potential zoonotic threat. In India alone it causes huge losses to stakeholders. Anthelmintics including triclabendazole have been used to control this menace but the emerging resistance against the available compounds necessitates identification of novel and alternative therapeutic measures involving plant derived natural compounds for their anthelmintic potential. Thymoquinone (T) and curcumin (C), the active ingredients of Nigella sativa and Curcuma longa respectively have been used as antiparasitic agents but the information on their flukicidal effect is very limited. Adult flukes of F. gigantica were in vitro exposed to different concentrations of thymoquinone and curcumin separately for 3h at 37+ 1°C. A significant (p<0.05) reduction in the worm motility at 60 μM concentration of both T and C was observed though all the worms remained alive after 3h exposure, whereas the effect on egg shedding was statistically insignificant. Pronounced tegumental disruptions and erosion of spines in the posterior region and around the acetabulum was evident. A significant (p<0.05) decrease in glutathione-S-transferase and superoxide dismutase activity and reduced glutathione (GSH) level was observed, while protein carbonylation increased differentially. A significant inhibition of CathepsinL (CatL) gene expression in thymoquinone treated worms was also evident. Further, in silico molecular docking of T and C with CatL revealed a stronger interaction of curcumin with the involvement of higher number of amino acids as compared to thymoquinone that could be more effective in inhibiting the antioxidant enzymes of F. gigantica. It is concluded that both the compounds understudy will decrease the detoxification ability of F. gigantica, while inhibition of CatL will significantly affect their virulence

  13. Immunolocalization and immunodetection of the excretory/secretory (ES) antigens of Fasciola gigantica.

    Science.gov (United States)

    Khan, M A Hannan; Ullah, Rizwan; Rehman, Abdur; Rehman, Lubna; P A, Ahammed Shareef; Abidi, S M A

    2017-01-01

    The digenetic trematode Fasciola gigantica is a parasite of great agricultural and economic importance. Along with Fasciola hepatica, F. gigantica incurs huge economic losses to the agricultural sector. Because of unavailability of an effective and commercial vaccine, the earliest diagnosis of the disease is the only way to control the disease. The conventional coprological techniques are able to detect the disease only after the parasites get matured and starts releasing their eggs with the faeces of host, therefore prepatent infection remain undiagnosed. The alternative method is by serological tests that uses circulatory antigens. Despite high sensitivity, their reliability is quite low because of the common antigens shared between different helminth parasites. To overcome this, investigation was shifted to identify the copro-antigens which could be more sensitive and reliable. In the present study, we tried to identify some of the immunodominant proteins from the Excretory Secretory (ES) product of F. gigantica which can be further characterized and used for early detection of infection and also as drug and vaccine candidates. The ES products of F. gigantica were collected and used for raising the polyclonal antibody in rabbit. The polypeptide profile was generated as well as immunogenic polypeptides were identified. The Source of ES antigen was immunolocalized using confocal microscopy and dot blot assay was performed to diagnose field infection. The polypeptide profile of ES products revealed a total of 24 polypeptides out of which 12 immunogenic polypeptides were identified by western blotting. Confocal micrographs showed the immunolocalization of antigens in the intestinal caecae, vitalline glands, gonads as well as in the tegument of the worm. The dot blot assay confirmed the utility of ES products for the detection of field infection. Subsequently, cross reactivity was found negative with Gigantocotyle explanatum; an amphitome parasite of same habitat

  14. Immunolocalization and immunodetection of the excretory/secretory (ES antigens of Fasciola gigantica.

    Directory of Open Access Journals (Sweden)

    M A Hannan Khan

    Full Text Available The digenetic trematode Fasciola gigantica is a parasite of great agricultural and economic importance. Along with Fasciola hepatica, F. gigantica incurs huge economic losses to the agricultural sector. Because of unavailability of an effective and commercial vaccine, the earliest diagnosis of the disease is the only way to control the disease. The conventional coprological techniques are able to detect the disease only after the parasites get matured and starts releasing their eggs with the faeces of host, therefore prepatent infection remain undiagnosed. The alternative method is by serological tests that uses circulatory antigens. Despite high sensitivity, their reliability is quite low because of the common antigens shared between different helminth parasites. To overcome this, investigation was shifted to identify the copro-antigens which could be more sensitive and reliable. In the present study, we tried to identify some of the immunodominant proteins from the Excretory Secretory (ES product of F. gigantica which can be further characterized and used for early detection of infection and also as drug and vaccine candidates. The ES products of F. gigantica were collected and used for raising the polyclonal antibody in rabbit. The polypeptide profile was generated as well as immunogenic polypeptides were identified. The Source of ES antigen was immunolocalized using confocal microscopy and dot blot assay was performed to diagnose field infection. The polypeptide profile of ES products revealed a total of 24 polypeptides out of which 12 immunogenic polypeptides were identified by western blotting. Confocal micrographs showed the immunolocalization of antigens in the intestinal caecae, vitalline glands, gonads as well as in the tegument of the worm. The dot blot assay confirmed the utility of ES products for the detection of field infection. Subsequently, cross reactivity was found negative with Gigantocotyle explanatum; an amphitome parasite

  15. Stimulating Neoblast-Like Cell Proliferation in Juvenile Fasciola hepatica Supports Growth and Progression towards the Adult Phenotype In Vitro

    Science.gov (United States)

    Rathinasamy, Vignesh; Toet, Hayley; McCammick, Erin; O’Connor, Anna; Marks, Nikki J.; Mousley, Angela; Brennan, Gerard P.; Halton, David W.; Spithill, Terry W.; Maule, Aaron G.

    2016-01-01

    Fascioliasis (or fasciolosis) is a socioeconomically important parasitic disease caused by liver flukes of the genus Fasciola. Flukicide resistance has exposed the need for new drugs and/or a vaccine for liver fluke control. A rapidly improving ‘molecular toolbox’ for liver fluke encompasses quality genomic/transcriptomic datasets and an RNA interference platform that facilitates functional genomics approaches to drug/vaccine target validation. The exploitation of these resources is undermined by the absence of effective culture/maintenance systems that would support in vitro studies on juvenile fluke development/biology. Here we report markedly improved in vitro maintenance methods for Fasciola hepatica that achieved 65% survival of juvenile fluke after 6 months in standard cell culture medium supplemented with 50% chicken serum. We discovered that this long-term maintenance was dependent upon fluke growth, which was supported by increased proliferation of cells resembling the “neoblast” stem cells described in other flatworms. Growth led to dramatic morphological changes in juveniles, including the development of the digestive tract, reproductive organs and the tegument, towards more adult-like forms. The inhibition of DNA synthesis prevented neoblast-like cell proliferation and inhibited growth/development. Supporting our assertion that we have triggered the development of juveniles towards adult-like fluke, mass spectrometric analyses showed that growing fluke have an excretory/secretory protein profile that is distinct from that of newly-excysted juveniles and more closely resembles that of ex vivo immature and adult fluke. Further, in vitro maintained fluke displayed a transition in their movement from the probing behaviour associated with migrating stage worms to a slower wave-like motility seen in adults. Our ability to stimulate neoblast-like cell proliferation and growth in F. hepatica underpins the first simple platform for their long-term in

  16. Cryopreservation and distribution of radiation-attenuated helminth larvae and the use of radioisotopes to monitor their survival. Coordinated programme on preparation of irradiated vaccines against some human diseases

    International Nuclear Information System (INIS)

    James, E.

    1982-06-01

    Techniques for the cryopreservation of schistosomula are described, from the methanol/two-step cooling technique, through a technique which uses 40% methanol and rapid cooling to the current technique which employs a two-step addition of ethanediol and rapid cooling. Levels of survival with these techniques have improved from 0.3% to 5.9% and now to 47% of control values. The 40% methanol/rapid cooling technique is described in detail as this forms the basis for understanding the role of cryoprotective additives and cooling and warming rates in the cryopreservation of schistosomula. The toxicity of 12 different potentially cryoprotective compounds is described. Cryopreservation of S.japonicum and S.bovis is described. The effect of the age of the schistosomula and their cryopreservability is related to the development of water sensitivity and the permeation and damage produced by glycerol and it is postulated that morphological changes occurring in the tegument during transformation from a cercaria to schistosomulum may account for these observations. Studies with 14 C-ethanediol are described which attempt to provide an understanding of permeability of cryoprtectants to schistosomula of different ages and at different temperatures. Vaccination studies with cryopreserved and radiation-attenuated schistosomula are also reported. Radiation-attenuated schistosomula are also reported. Radiation-attenuated cercariae and schistosomula produced high (64% to 89%) levels of protection in baboons, and cryopreserved schistosomula produced comparable levels of protection in vaccinated mice to normal schistosomula. Cryopreserved radiation-attenuated schistosomula produced a significant level of protection (49% reduction) in sheep although the numbers of normally motile organisms injected was low (1,000 per dose, 2 doses). It is concluded that normally-motile cryopreserved radiation-attenuated schistosomula are as immunogenic as fresh organisms

  17. Aspectos morfológicos de frutos e sementes e caracterização citogenética de Crotalaria lanceolata E. Mey. (Papilionoideae - Fabaceae Morphological aspects of fruits and seeds and cytogenetic characterization of Crotalaria lanceolata E. Mey. (Papilionoideae - Fabaceae

    Directory of Open Access Journals (Sweden)

    Débora Aparecida Verde de Andrade

    2008-09-01

    Full Text Available Frutos, sementes e plântulas de Crotalaria lanceolata, conhecida popularmente como guizo-de-cascavel, chocalho-de-cobra, xique-xique ou feijão-de-guizo, planta tóxica infestante que ocorre no Estado de São Paulo, foram estudadas morfologicamente e citogeneticamente. Os frutos são secos, deiscentes, polispérmicos e do tipo legume. As sementes são reniformes e o embrião é constituído de eixo embrionário e dois cotilédones. A testa pode apresentar variadas tonalidades de castanhos. A germinação é epígea e fanerocotiledonar. A espécie apresenta número cromossômico diplóide 2n = 16 com formulação cariotípica 12M + 4SM e comprimento cromossômico médio geral de 3,340 ± 0,689.Crotalaria lanceolata, known popularly as "guizo-de-cascavel", "xique-xique" or "feijão-de-guizo", is a toxic weed that occurs in the state of Sao Paulo. Fruits, seeds and seedlings were studied morphologically and cytogenetically. The fruits are dry, dehiscent, polyspermic legumes. The seeds are kidney shaped and the embryo consists of an embryonic axis and two cotyledons. The seed tegument comes in various chestnut tones. Germination is phanerocotylar and epigeal. The species has a mitotic chromosome number 2n = 16 with karyotype formula 12M + 4SM; overall mean chromosome length is 3.340 ± 0.689.

  18. The morphology and attachment of Protopolystoma xenopodis (Monogenea: Polystomatidae infecting the African clawed frog Xenopus laevis

    Directory of Open Access Journals (Sweden)

    Theunissen Maxine

    2014-01-01

    Full Text Available The African clawed frog Xenopus laevis (Anura: Pipidae is host to more than 25 parasite genera encompassing most of the parasitic invertebrate groups. Protopolystoma xenopodis Price, 1943 (Monogenea: Polystomatidae is one of two monogeneans infecting X. laevis. This study focussed on the external morphology of different developmental stages using scanning electron microscopy, histology and light microscopy. Eggs are released continuously and are washed out when the frog urinates. After successful development, an active swimming oncomiracidium leaves the egg capsule and locates a potential post-metamorphic clawed frog. The oncomiracidium migrates to the kidney where it attaches and starts to feed on blood. The parasite then migrates to the urinary bladder where it reaches maturity. Eggs are fusiform, about 300 μm long, with a smooth surface and are operculated. Oncomiracidia are elongated and cylindrical in shape, with an oval posterior cup-shaped haptor that bears a total of 20 sclerites; 16 marginal hooklets used for attachment to the kidney of the host and two pairs of hamulus primordia. Cilia from the 64 ciliated cells enable the oncomiracidium to swim for up to 24 h when the cilia subsequently curl up, become non-functional and are shed from the body. The tegument between the ciliated cells bears a series of sensory papillae. The body of the mature parasite is elongated and pyriform and possesses an opisthaptor armed with three pairs of suckers and two pairs of falciform hooks to ensure a firm grip on the flexible internal surface of the urinary bladder.

  19. Characterization of a gene family encoding SEA (sea-urchin sperm protein, enterokinase and agrin-domain proteins with lectin-like and heme-binding properties from Schistosoma japonicum.

    Directory of Open Access Journals (Sweden)

    Evaristus Chibunna Mbanefo

    Full Text Available BACKGROUND: We previously identified a novel gene family dispersed in the genome of Schistosoma japonicum by retrotransposon-mediated gene duplication mechanism. Although many transcripts were identified, no homolog was readily identifiable from sequence information. METHODOLOGY/PRINCIPAL FINDINGS: Here, we utilized structural homology modeling and biochemical methods to identify remote homologs, and characterized the gene products as SEA (sea-urchin sperm protein, enterokinase and agrin-domain containing proteins. A common extracellular domain in this family was structurally similar to SEA-domain. SEA-domain is primarily a structural domain, known to assist or regulate binding to glycans. Recombinant proteins from three members of this gene family specifically interacted with glycosaminoglycans with high affinity, with potential implication in ligand acquisition and immune evasion. Similar approach was used to identify a heme-binding site on the SEA-domain. The heme-binding mode showed heme molecule inserted into a hydrophobic pocket, with heme iron putatively coordinated to two histidine axial ligands. Heme-binding properties were confirmed using biochemical assays and UV-visible absorption spectroscopy, which showed high affinity heme-binding (K D = 1.605×10(-6 M and cognate spectroscopic attributes of hexa-coordinated heme iron. The native proteins were oligomers, antigenic, and are localized on adult worm teguments and gastrodermis; major host-parasite interfaces and site for heme detoxification and acquisition. CONCLUSIONS: The results suggest potential role, at least in the nucleation step of heme crystallization (hemozoin formation, and as receptors for heme uptake. Survival strategies exploited by parasites, including heme homeostasis mechanism in hemoparasites, are paramount for successful parasitism. Thus, assessing prospects for application in disease intervention is warranted.

  20. Molecular characterization of severin from Clonorchis sinensis excretory/secretory products and its potential anti-apoptotic role in hepatocarcinoma PLC cells.

    Directory of Open Access Journals (Sweden)

    Xueqing Chen

    Full Text Available BACKGROUND: Clonorchiasis, caused by the infection of Clonorchis sinensis (C. sinensis, is a kind of neglected tropical disease, but it is highly related to cholangiocarcinoma and hepatocellular carcinoma (HCC. It has been well known that the excretory/secretory products of C. sinensis (CsESPs play key roles in clonorchiasis associated carcinoma. From genome and transcriptome of C. sinensis, we identified one component of CsESPs, severin (Csseverin, which had three putative gelsolin domains. Its homologues are supposed to play a vital role in apoptosis resistance of tumour cell. METHODOLOGY/PRINCIPAL FINDINGS: There was significant similarity in tertiary structures between human gelsolin and Csseverin by bioinformatics analysis. We identified that Csseverin expressed at life stage of adult worm, metacercaria and egg by the method of quantitative real-time PCR and western blotting. Csseverin distributed in vitellarium and intrauterine eggs of adult worm and tegument of metacercaria by immunofluorence assay. We obtained recombinant Csseverin (rCsseverin and confirmed that rCsseverin could bind with calciumion in circular dichroism spectrum analysis. It was demonstrated that rCsseverin was of the capability of actin binding by gel overlay assay and immunocytochemistry. Both Annexin V/PI assay and mitochondrial membrane potential assay of human hepatocarcinoma cell line PLC showed apoptosis resistance after incubation with different concentrations of rCsseverin. Morphological analysis, apoptosis-associated changes of mitochondrial membrane potential and Annexin V/PI apoptosis assay showed that co-incubation of PLC cells with rCsseverin in vitro led to an inhibition of apoptosis induced by serum-starved for 24 h. CONCLUSIONS/SIGNIFICANCE: Collectively, the molecular properties of Csseverin, a molecule of CsESPs, were characterized in our study. rCsseverin could cause obvious apoptotic inhibition in human HCC cell line. Csseverin might exacerbate the

  1. Molecular Characterization of Severin from Clonorchis sinensis Excretory/Secretory Products and Its Potential Anti-apoptotic Role in Hepatocarcinoma PLC Cells

    Science.gov (United States)

    He, Lei; Wang, Xiaoyun; Liang, Pei; Chen, Wenjun; Bian, Meng; Ren, Mengyu; Lin, Jinsi; Liang, Chi; Xu, Jin; Wu, Zhongdao; Li, Xuerong; Huang, Yan; Yu, Xinbing

    2013-01-01

    Background Clonorchiasis, caused by the infection of Clonorchis sinensis (C. sinensis), is a kind of neglected tropical disease, but it is highly related to cholangiocarcinoma and hepatocellular carcinoma (HCC). It has been well known that the excretory/secretory products of C. sinensis (CsESPs) play key roles in clonorchiasis associated carcinoma. From genome and transcriptome of C. sinensis, we identified one component of CsESPs, severin (Csseverin), which had three putative gelsolin domains. Its homologues are supposed to play a vital role in apoptosis resistance of tumour cell. Methodology/Principal Findings There was significant similarity in tertiary structures between human gelsolin and Csseverin by bioinformatics analysis. We identified that Csseverin expressed at life stage of adult worm, metacercaria and egg by the method of quantitative real-time PCR and western blotting. Csseverin distributed in vitellarium and intrauterine eggs of adult worm and tegument of metacercaria by immunofluorence assay. We obtained recombinant Csseverin (rCsseverin) and confirmed that rCsseverin could bind with calciumion in circular dichroism spectrum analysis. It was demonstrated that rCsseverin was of the capability of actin binding by gel overlay assay and immunocytochemistry. Both Annexin V/PI assay and mitochondrial membrane potential assay of human hepatocarcinoma cell line PLC showed apoptosis resistance after incubation with different concentrations of rCsseverin. Morphological analysis, apoptosis-associated changes of mitochondrial membrane potential and Annexin V/PI apoptosis assay showed that co-incubation of PLC cells with rCsseverin in vitro led to an inhibition of apoptosis induced by serum-starved for 24 h. Conclusions/Significance Collectively, the molecular properties of Csseverin, a molecule of CsESPs, were characterized in our study. rCsseverin could cause obvious apoptotic inhibition in human HCC cell line. Csseverin might exacerbate the process of HCC

  2. Comparison of two serpins of Clonorchis sinensis by bioinformatics, expression, and localization in metacercaria.

    Science.gov (United States)

    Yang, Yabo; Hu, Dong; Wang, Lexun; Liang, Chi; Hu, Xuchu; Xu, Jin; Huang, Yan; Yu, Xinbing

    2014-06-01

    Clonorchiasis, which has been an important public health problem in China, is caused by ingestion of raw or undercooked fish contaminated by live metacercaria. Therefore, preventing fish from infecting is of great significance for controlling the disease. SERPINs (serine protease inhibitors) are well known as negative regulators of hemostasis, thrombolysis, and innate immune responses. In the present study, two full-length sequences encoding SERPIN were identified from metacercaria cDNA library of Clonorchis sinensis (C. sinensis) and were denominated as CsSERPIN and CsSERPIN3, respectively. Bioinformatics analysis showed that the two sequences shares 35.9% identity to each other. Both of the sequences have SERPIN domain and the greatest difference between the two domains is the reactive centre loop. Transmembrane region was found in CsSERPIN3 while not in CsSERPIN. The expression of the two CsSERPINs was significantly higher at the life stage of metacercaria than that of adult. The transcription levels of CsSERPIN and CsSERPIN3 at metacercaria stage were 3.249- and 11.314-fold of that at adult stage, respectively. Furthermore, the expression of CsSERPIN was 4.32-fold of that of CsSERPIN3 at metacercaria stage. Immunobiochemistry revealed that CsERPIN was dispersed at subtegument and oral sucker of metacercaria, while CsSERPIN3 localized intensely in the tegument of metacercaria of C. sinensis inside of the cyst wall. All these indicated that the CsSERPINs play important roles at metacercaria stage of the parasite. CsSERPIN may take part in regulation of endogenous serine proteinase and CsSERPIN3 may be involved in immune evasion and be a potential candidate for vaccine and drug target for clonorchiasis.

  3. In vitro activity of Origanum vulgare essential oil against Candida species

    Directory of Open Access Journals (Sweden)

    Marlete Brum Cleff

    2010-03-01

    Full Text Available The aim of this study was to evaluate the in vitro activity of the essential oil extracted from Origanum vulgare against sixteen Candida species isolates. Standard strains tested comprised C. albicans (ATCC strains 44858, 4053, 18804 and 3691, C. parapsilosis (ATCC 22019, C. krusei (ATCC 34135, C. lusitaniae (ATCC 34449 and C. dubliniensis (ATCC MY646. Six Candida albicans isolates from the vaginal mucous membrane of female dogs, one isolate from the cutaneous tegument of a dog and one isolate of a capuchin monkey were tested in parallel. A broth microdilution technique (CLSI was used, and the inoculum concentration was adjusted to 5 x 10(6 CFU mL-1. The essential oil was obtained by hydrodistillation in a Clevenger apparatus and analyzed by gas chromatography. Susceptibility was expressed as Minimal Inhibitory Concentration (MIC and Minimal Fungicidal Concentration (MFC. All isolates tested in vitro were sensitive to O. vulgare essential oil. The chromatographic analysis revealed that the main compounds present in the essential oil were 4-terpineol (47.95%, carvacrol (9.42%, thymol (8.42% and □-terpineol (7.57%. C. albicans isolates obtained from animal mucous membranes exhibited MIC and MFC values of 2.72 µL mL-1 and 5 µL mL-1, respectively. MIC and MFC values for C. albicans standard strains were 2.97 µL mL-1 and 3.54 µL mL-1, respectively. The MIC and MFC for non-albicans species were 2.10 µL mL-1 and 2.97 µL mL-1, respectively. The antifungal activity of O. vulgare essential oil against Candida spp. observed in vitro suggests its administration may represent an alternative treatment for candidiasis.

  4. A NEW SPECIES AND A COMBINATION IN CARDIOSPERMUM (SAPINDACEAE

    Directory of Open Access Journals (Sweden)

    María Silvia Ferrucci

    1993-01-01

    Full Text Available Cardiospermum heringeri Ferrucci, a new species from Brazil isdescribed and illustrated. It shares the sect. Ceratadenia ·RadJk.· with C. grandiflorum Sw. and C. integerrimum Radlk., a key for recognizing them is given. C. integerrimum is closely related with the new taxa, observations on the morphology of seed tegument and pollen of both species, complement this study.Based on Paullinia urvilleoides Radlk., the new combinationCardiospermum urvilleoides (Radlk. Ferrucci is established; it belongs to the sect. Cardiospermum. Fruit characters and a detailed palynological description provides evidence in favour of its removal from Paullinia L. Cardiospermum L. comparte la subtribu Paulliniinae Radlk. con los géneros Serjania Miller, Houssayanthus Hunz., Paullinia L., Urvillea Kunth y Lophostigma Radlk. Los caracteres comunes a todos los géneros son las flores zigomorfas, con las escamas de los pétalos en forma de capucha, y los frutos, que pueden ser esquizocárpicos o bien cápsulas. Dentro de la subtribu, Cardiospermum es el género más heterogéneo e interesantepor la variabilidad morfológica y cromosómica.El estudio de una colección de Sapindáceas procedentes de Brasil (Bahia, Minas Gerais y Espírito Santo, permite presentar una especie nueva para la ciencia: C. heringeri Ferrucci que en este trabajo se comparacon la especie afín C. integerrimum Radlk. También se establece una nuevacombinación en la sección Cardiospermum

  5. Inhibition or knockdown of ABC transporters enhances susceptibility of adult and juvenile schistosomes to Praziquantel.

    Directory of Open Access Journals (Sweden)

    Ravi S Kasinathan

    2014-10-01

    Full Text Available Parasitic flatworms of the genus Schistosoma cause schistosomiasis, a neglected tropical disease that affects hundreds of millions. Treatment of schistosomiasis depends almost entirely on the drug praziquantel (PZQ. Though essential to treating and controlling schistosomiasis, a major limitation of PZQ is that it is not active against immature mammalian-stage schistosomes. Furthermore, there are reports of field isolates with heritable reductions in PZQ susceptibility, and researchers have selected for PZQ-resistant schistosomes in the laboratory. P-glycoprotein (Pgp; ABCB1 and other ATP binding cassette (ABC transporters remove a wide variety of toxins and xenobiotics from cells, and have been implicated in multidrug resistance (MDR. Changes in ABC transporter structure or expression levels are also associated with reduced drug susceptibility in parasitic helminths, including schistosomes. Here, we show that the activity of PZQ against schistosome adults and juveniles ex vivo is potentiated by co-administration of either the highly potent Pgp inhibitor tariquidar or combinations of inhibitors targeting multiple ABC multidrug transporters. Adult worms exposed to sublethal PZQ concentrations remain active, but co-administration of ABC transporter inhibitors results in complete loss of motility and disruption of the tegument. Notably, juvenile schistosomes (3-4 weeks post infection, normally refractory to 2 µM PZQ, become paralyzed when transporter inhibitors are added in combination with the PZQ. Experiments using the fluorescent PZQ derivative (R-PZQ-BODIPY are consistent with the transporter inhibitors increasing effective intraworm concentrations of PZQ. Adult worms in which expression of ABC transporters has been suppressed by RNA interference show increased responsiveness to PZQ and increased retention of (R-PZQ-BODIPY consistent with an important role for these proteins in setting levels of PZQ susceptibility. These results indicate that

  6. BinAligner: a heuristic method to align biological networks.

    Science.gov (United States)

    Yang, Jialiang; Li, Jun; Grünewald, Stefan; Wan, Xiu-Feng

    2013-01-01

    The advances in high throughput omics technologies have made it possible to characterize molecular interactions within and across various species. Alignments and comparison of molecular networks across species will help detect orthologs and conserved functional modules and provide insights on the evolutionary relationships of the compared species. However, such analyses are not trivial due to the complexity of network and high computational cost. Here we develop a mixture of global and local algorithm, BinAligner, for network alignments. Based on the hypotheses that the similarity between two vertices across networks would be context dependent and that the information from the edges and the structures of subnetworks can be more informative than vertices alone, two scoring schema, 1-neighborhood subnetwork and graphlet, were introduced to derive the scoring matrices between networks, besides the commonly used scoring scheme from vertices. Then the alignment problem is formulated as an assignment problem, which is solved by the combinatorial optimization algorithm, such as the Hungarian method. The proposed algorithm was applied and validated in aligning the protein-protein interaction network of Kaposi's sarcoma associated herpesvirus (KSHV) and that of varicella zoster virus (VZV). Interestingly, we identified several putative functional orthologous proteins with similar functions but very low sequence similarity between the two viruses. For example, KSHV open reading frame 56 (ORF56) and VZV ORF55 are helicase-primase subunits with sequence identity 14.6%, and KSHV ORF75 and VZV ORF44 are tegument proteins with sequence identity 15.3%. These functional pairs can not be identified if one restricts the alignment into orthologous protein pairs. In addition, BinAligner identified a conserved pathway between two viruses, which consists of 7 orthologous protein pairs and these proteins are connected by conserved links. This pathway might be crucial for virus packing and

  7. submetidas a diferentes condições de armazenamento e a escarificação

    Directory of Open Access Journals (Sweden)

    Álvaro Valente Caçola

    2006-01-01

    Full Text Available The seeds of Araucaria angustifolia (Bertol. Kuntze are recalcitrants and, therefore, they lose rapidly the viability after the harvest, limiting their use for nursery production of seedlings. This work was carried out to investigate the effects of cold storage duration, conservation method, and escarification on germination and vigour of seeds of Araucaria angustifolia. The seeds were cold stored (0-1oC/90-95% RH for 0, 60, 120, and 180 days, under normal air condictions (NA, modified atmosphere (MA, and controlled atmosphere (CA storage. For each cold storage duration, after removal from cold storage, the seeds were submitted or not to scarification (by cutting a small portion of the apical seed tegument, avoiding the damage of the endosperm, letting the germinate in a chamber with controlled environment, in plastic trays filled with vermiculite, for 60 days. The experiment followed the completely randomized factorial design (4x3x2, with four cold storage durations (0, 60, 120, and 180 days, three storage methods (NA, MA, and CA, with or without escarification, and four replicates. There was a good preservation of physiological quality for seeds submitted to different conservation methods in cold storage along the 180-day period. However, there was a faster germination and initial growth of seedlings for seeds left in cold storage for 60 days than in seeds assessed at harvest or left in cold storage for 120-180 days. The NA storage of seeds in perfurated plastic bags was as efficient as MA and CA storage to preserve seeds quality. Seeds scarification increased vigor and promoted seedlings initial growth of Araucaria angustifolia after being removed from cold storage.

  8. New 1,3-benzodioxole derivatives: Synthesis, evaluation of in vitro schistosomicidal activity and ultrastructural analysis.

    Science.gov (United States)

    Mariz Gomes da Silva, Luana Maria; de Oliveira, Jamerson Ferreira; Silva, Willams Leal; da Silva, Anekecia Lauro; de Almeida Junior, Antônio Sérgio Alves; Barbosa Dos Santos, Victor Hugo; Alves, Luiz Carlos; Brayner Dos Santos, Fábio André; Costa, Vlaudia Maria Assis; Aires, André de Lima; de Lima, Maria do Carmo Alves; Albuquerque, Monica Camelo Pessoa de Azevedo

    2018-03-01

    Schistosomiasis is considered a serious public health problem in 78 countries and territories located in Africa, Asia and America and it is estimated in more than 249 million people infected by any of the species of Schistosoma. The exclusive use of praziquantel (PZQ), effective drug against all species of Schistosoma, has been the basis of the development of a possible resistance against the strains of this parasite. In addition, PZQ is not effective against young forms of worms. Thus, there is a need for the development of new drugs with schistosomicidal activity. The objective of this work was to synthesize and to evaluate the therapeutic potential of new benzodioxole derivatives (3-14) candidates for schistosomicidal drugs. All compounds synthesized showed in vitro schistosomicidal activity. The derivative 12 was considered the best compound, since it took 100% of worms to mortality in the first 72 h of exposure at the concentration of 100 μM and 83.3% at the concentration of 50 μM. Furthermore, male and female adult worms, incubated for 24 h with the compound 12 showed tegument damages characterized by extensive desquamation and edema, tuber destruction, bubble formation and exposure of the muscle layer. This compound has a restricted structure, where the thiazolidinone is attached to the 4-position of the 1,3-benzodioxol ring. The structural conformation of derivative 12 was probably responsible for the promising schistosomicidal activity, where the presence of an electron/conformational restriction of the thiazolidine ring, as well as the action of bromine as a bulk substitute, favored an increase in biological activity. In addition, tegumentary changes caused by derivative 12 may also have been responsible for the death of adult worms of Schistosoma mansoni. Therefore, we verified that the results obtained in this study make benzodioxole derivatives possible candidates for prototypes of new schistosomicidal drugs. Copyright © 2018 Elsevier B

  9. In-vitro replication of Chelonid herpesvirus 5 in organotypic skin cultures from Hawaiian green turtles (Chelonia mydas)

    Science.gov (United States)

    Work, Thierry M.; Dagenais, Julie; Weatherby, Tina; Ackermann, Mathias; Balazs, George H.

    2017-01-01

    Fibropapillomatosis (FP) is a tumor disease of marine turtles associated with Chelonid herpesvirus 5 (ChHV5) that has historically been refractory to growth in tissue culture. Here, we show for the first time de novo formation of ChHV5-positive intranuclear inclusions in cultured green turtle cells, which is indicative for active lytic replication of the virus. The minimal requirements to achieve lytic replication in cultured cells included 1) either in-vitro culturing of ChHV5-positive tumor biopsies (plugs) or organotypic cultures (rafts) consisting of ChHV5-positive turtle fibroblasts in collagen rafts seeded with turtle keratinocytes and 2) keratinocyte maturation induced by raising raft or biopsy cultures to the air-liquid interface. Virus growth was confirmed by detailed electron microscopic studies revealing intranuclear sun-shaped capsid factories, tubules, various stages of capsid formation, nuclear export by budding into the perinuclear space, tegumentation, and envelopment to complete de novo virus production. Membrane synthesis was also observed as a sign for active viral replication. Interestingly, cytoplasmic particles became associated with keratin filaments, a feature not seen in conventional monolayer cell cultures where most studies of herpesvirus replication have been performed. Our findings draw a rich and realistic picture of ChHV5 replication in cells derived from its natural host and may be crucial not only to better understand ChHV5 circulation but also to eventually complete Koch's postulates for FP. Moreover, the principles described here may serve as model to culture other viruses that are resistant to replication in conventional cell culture.

  10. Herpesvirus capsid assembly and DNA packaging

    Science.gov (United States)

    Heming, Jason D.; Conway, James F.; Homa, Fred L.

    2017-01-01

    Herpes simplex virus type I (HSV-1) is the causative agent of several pathologies ranging in severity from the common cold sore to life-threatening encephalitic infection. During productive lytic infection, over 80 viral proteins are expressed in a highly regulated manner, resulting in the replication of viral genomes and assembly of progeny virions. The virion of all herpesviruses consists of an external membrane envelope, a proteinaceous layer called the tegument, and an icosahedral capsid containing the double-stranded linear DNA genome. The capsid shell of HSV-1 is built from four structural proteins: a major capsid protein, VP5, which forms the capsomers (hexons and pentons), the triplex consisting of VP19C and VP23 found between the capsomers, and VP26 which binds to VP5 on hexons but not pentons. In addition, the dodecameric pUL6 portal complex occupies one of the 12 capsid vertices, and the capsid vertex specific component (CVSC), a heterotrimer complex of pUL17, pUL25 and pUL36 binds specifically to the triplexes adjacent to each penton. The capsid is assembled in the nucleus where the viral genome is packaged into newly assembled closed capsid shells. Cleavage and packaging of replicated, concatemeric viral DNA requires the seven viral proteins encoded by the UL6, UL15, UL17, UL25, UL28, UL32, and UL33 genes. Considerable advances have been made in understanding the structure of the herpesvirus capsid and the function of several of the DNA packaging proteins by applying biochemical, genetic, and structural techniques. This review is a summary of recent advances with respect to the structure of the HSV-1 virion capsid and what is known about the function of the seven packaging proteins and their interactions with each other and with the capsid shell. PMID:28528442

  11. Novel therapeutic and prevention approaches for schistosomiasis: review.

    Science.gov (United States)

    El Ridi, Rashika A F; Tallima, Hatem A-M

    2013-09-01

    Schistosomiasis is a debilitating disease affecting approximately 600 million people in 74 developing countries, with 800 million, mostly children at risk. To circumvent the threat of having praziquantel (PZQ) as the only drug used for treatment, several PZQ derivatives were synthesized, and drugs destined for other parasites were used with success. A plethora of plant-derived oils and extracts were found to effectively kill juvenile and adult schistosomes, yet none was progressed to pre- and clinical studies except an oleo-gum resin extracted from the stem of Commiphora molmol, myrrh, which action was challenged in several trials. We have proposed an essential fatty acid, a component of our diet and cells, the polyunsaturated fatty acid arachidonic acid (ARA) as a remedy for schistosomiasis, due to its ability to activate the parasite tegument-bound neutral sphingomyelinase, with subsequent hydrolysis of the apical lipid bilayer sphingomyelin molecules, allowing access of specific antibody molecules, and eventual worm attrition. This concept was convincingly supported using larval and adult Schistosoma mansoni and Schistosoma haematobium worms in in vitro experiments, and in vivo studies in inbred mice and outbred hamsters. Even if ARA proves to be an entirely effective and safe therapy for schistosomiasis, it will not prevent reinfection, and accordingly, the need for developing an effective vaccine remains an urgent priority. Our studies have supported the status of S. mansoni calpain, glutathione-S-transferase, aldolase, triose phosphate isomerase, glyceraldehyde 3-phosphate dehydrogenase, enolase, and 2-cys peroxiredoxin as vaccine candidates, as they are larval excreted-secreted products and, contrary to the surface membrane molecules, are entirely accessible to the host immune system effector elements. We have proposed that the use of these molecules, in conjunction with Th2 cytokines-inducing adjuvants for recruiting and activating eosinophils and basophils

  12. Novel Therapeutic and Prevention Approaches for Schistosomiasis: Review

    Directory of Open Access Journals (Sweden)

    Rashika A.F. El Ridi

    2013-09-01

    Full Text Available Schistosomiasis is a debilitating disease affecting approximately 600 million people in 74 developing countries, with 800 million, mostly children at risk. To circumvent the threat of having praziquantel (PZQ as the only drug used for treatment, several PZQ derivatives were synthesized, and drugs destined for other parasites were used with success. A plethora of plant-derived oils and extracts were found to effectively kill juvenile and adult schistosomes, yet none was progressed to pre- and clinical studies except an oleo-gum resin extracted from the stem of Commiphora molmol, myrrh, which action was challenged in several trials. We have proposed an essential fatty acid, a component of our diet and cells, the polyunsaturated fatty acid arachidonic acid (ARA as a remedy for schistosomiasis, due to its ability to activate the parasite tegument-bound neutral sphingomyelinase, with subsequent hydrolysis of the apical lipid bilayer sphingomyelin molecules, allowing access of specific antibody molecules, and eventual worm attrition. This concept was convincingly supported using larval and adult Schistosoma mansoni and Schistosoma haematobium worms in in vitro experiments, and in vivo studies in inbred mice and outbred hamsters. Even if ARA proves to be an entirely effective and safe therapy for schistosomiasis, it will not prevent reinfection, and accordingly, the need for developing an effective vaccine remains an urgent priority. Our studies have supported the status of S. mansoni calpain, glutathione-S-transferase, aldolase, triose phosphate isomerase, glyceraldehyde 3-phosphate dehydrogenase, enolase, and 2-cys peroxiredoxin as vaccine candidates, as they are larval excreted-secreted products and, contrary to the surface membrane molecules, are entirely accessible to the host immune system effector elements. We have proposed that the use of these molecules, in conjunction with Th2 cytokines-inducing adjuvants for recruiting and activating

  13. The nature and combination of subunits used in epitope-based Schistosoma japonicum vaccine formulations affect their efficacy

    Directory of Open Access Journals (Sweden)

    Liu Feng

    2010-11-01

    Full Text Available Abstract Background Schistosomiasis remains a major public health problem in endemic countries and is caused by infections with any one of three primary schistosome species. Although there are no vaccines available to date, this strategy appears feasible since natural immunity develops in individuals suffering from repeated infection during a lifetime. Since vaccinations resulting in both Th1- and Th2-type responses have been shown to contribute to protective immunity, a vaccine formulation with the capacity for stimulating multiple arms of the immune response will likely be the most effective. Previously we developed partially protective, single Th- and B cell-epitope-based peptide-DNA dual vaccines (PDDV (T3-PDDV and B3-PDDV, respectively capable of eliciting immune responses against the Schistosoma japonicum 22.6 kDa tegument antigen (Sj22.6 and a 62 kDa fragment of myosin (Sj62, respectively. Results In this study, we developed PDDV cocktails containing multiple epitopes of S. japonicum from Sj22.6, Sj62 and Sj97 antigens by predicting cytotoxic, helper, and B-cell epitopes, and evaluated vaccine potential in vivo. Results showed that mice immunized with a single-epitope PDDV elicited either Tc, Th, or B cell responses, respectively, and mice immunized with either the T3- or B3- single-epitope PDDV formulation were partially protected against infection. However, mice immunized with a multicomponent (3 PDDV components formulation elicited variable immune responses that were less immunoprotective than single-epitope PDDV formulations. Conclusions Our data show that combining these different antigens did not result in a more effective vaccine formulation when compared to each component administered individually, and further suggest that immune interference resulting from immunizations with antigenically distinct vaccine targets may be an important consideration in the development of multicomponent vaccine preparations.

  14. Psettarium anthicum sp. n. (Digenea: Sanguinicolidae) from the heart of cobia Rachycentron canadum (Rachycentridae) in the northern Gulf of Mexico.

    Science.gov (United States)

    Bullard, Stephen A; Overstreet, Robin M

    2006-06-01

    Psettarium anthicum sp. n. (Digenea: Sanguinicolidae) infects the myocardium and atrial wall of the cobia Rachycentron canadum (Linnaeus, 1766) (Rachycentridae) in the northern Gulf of Mexico off Mississippi, USA. It is the first member of Psettarium Goto et Ozaki, 1930 reported from other than the Indian Ocean or Pacific Ocean and the second species of the genus reported from cobia. It differs from its congeners by the combination of having posterior caeca with lateral projections appearing as thorns in lateral view and the male pore anterior to the oötype. The species of Psettarium, P. japonicum (Goto et Ozaki, 1929) (type species), P. tropicum Manter, 1940, P. sebastodorum Holmes, 1971, P. rachycentri (Lebedev et Parukhin, 1972) comb. n. (syn. Psettarioides rachycentri Lebedev et Parukhin, 1972) and P. anthicum sp. n., differ from other sanguinicolids by the combination of having an elongate body with a sinistral posterolateral protuberance, minute, straight tegumental body spines in ventrolateral transverse rows, posterior caeca greater than seven time the anterior caeca length, the o6type near the posterior end of the body, a uterus primarily between the ovary and oötype and an oviduct and vitelline duct extending posteriad primarily between the uterus and dextral body margin. We emend Psettarium and provide a diagnostic key to the species. Psettarioides is regarded as a junior synonym of Psettarium because herein we return its type species, P. tropicum, to Psettarium. Regarding the three other sanguinicolids formerly of Psettarioides, we suspect that P. pseudupenei Lebedev et Parukhin, 1972 belongs to Psettarium but include it only tentatively pending an examination of type or other material; we tentatively place P. kurochkini Parukhin, 1976 in Cardicola Short, 1952; and we designate P. grandis (Lebedev et Mamaev, 1968) as incertae sedis pending examination of type or other appropriate material.

  15. Direct interaction between EgFABP1, a fatty acid binding protein from Echinococcus granulosus, and phospholipid membranes.

    Directory of Open Access Journals (Sweden)

    Jorge L Porfido

    Full Text Available Growth and maintenance of hydatid cysts produced by Echinococcus granulosus have a high requirement for host lipids for biosynthetic processes, membrane building and possibly cellular and developmental signalling. This requires a high degree of lipid trafficking facilitated by lipid transporter proteins. Members of the fatty acid binding protein (FABP family have been identified in Echinococcus granulosus, one of which, EgFABP1 is expressed at the tegumental level in the protoscoleces, but it has also been described in both hydatid cyst fluid and secretions of protoscoleces. In spite of a considerable amount of structural and biophysical information on the FABPs in general, their specific functions remain mysterious.We have investigated the way in which EgFABP1 may interact with membranes using a variety of fluorescence-based techniques and artificial small unilamellar vesicles. We first found that bacterial recombinant EgFABP1 is loaded with fatty acids from the synthesising bacteria, and that fatty acid binding increases its resistance to proteinases, possibly due to subtle conformational changes induced on EgFABP1. By manipulating the composition of lipid vesicles and the ionic environment, we found that EgFABP1 interacts with membranes in a direct contact, collisional, manner to exchange ligand, involving both ionic and hydrophobic interactions. Moreover, we observed that the protein can compete with cytochrome c for association with the surface of small unilamellar vesicles (SUVs.This work constitutes a first approach to the understanding of protein-membrane interactions of EgFABP1. The results suggest that this protein may be actively involved in the exchange and transport of fatty acids between different membranes and cellular compartments within the parasite.

  16. Cell-cycle-dependent localization of human cytomegalovirus UL83 phosphoprotein in the nucleolus and modulation of viral gene expression in human embryo fibroblasts in vitro.

    Science.gov (United States)

    Arcangeletti, Maria-Cristina; Rodighiero, Isabella; Mirandola, Prisco; De Conto, Flora; Covan, Silvia; Germini, Diego; Razin, Sergey; Dettori, Giuseppe; Chezzi, Carlo

    2011-01-01

    The nucleolus is a multifunctional nuclear compartment widely known to be involved in several cellular processes, including mRNA maturation and shuttling to cytoplasmic sites, control of the cell cycle, cell proliferation, and apoptosis; thus, it is logical that many viruses, including herpesvirus, target the nucleolus in order to exploit at least one of the above-mentioned functions. Recent studies from our group demonstrated the early accumulation of the incoming ppUL83 (pp65), the major tegument protein of human cytomegalovirus (HCMV), in the nucleolus. The obtained results also suggested that a functional relationship might exist between the nucleolar localization of pp65, rRNA synthesis, and the development of the lytic program of viral gene expression. Here we present new data which support the hypothesis of a potentially relevant role of HCMV pp65 and its nucleolar localization for the control of the cell cycle by HCMV (arrest of cell proliferation in G1-G1/S), and for the promotion of viral infection. We demonstrated that, although the incoming pp65 amount in the infected cells appears to be constant irrespective of the cell-cycle phase, its nucleolar accumulation is prominent in G1 and G1/S, but very poor in S or G2/M. This correlates with the observation that only cells in G1 and G1/S support an efficient development of the HCMV lytic cycle. We propose that HCMV pp65 might be involved in regulatory/signaling pathways related to nucleolar functions, such as the cell-cycle control. Co-immunoprecipitation experiments have permitted to identify nucleolin as one of the nucleolar partners of pp65.

  17. Development of Two Antibody Detection Enzyme-Linked Immunosorbent Assays for Serodiagnosis of Human Chronic Fascioliasis

    Science.gov (United States)

    Cabán-Hernández, Kimberly; Gaudier, José F.; Ruiz-Jiménez, Caleb

    2014-01-01

    Coprological examination based on egg detection in stool samples is currently used as the gold standard for the diagnosis of human fascioliasis. However, this method is not effective during the acute phase of the disease and has poor sensitivity during the chronic phase. Serodiagnosis has become an excellent alternative to coprological examination in efforts to combat the effects of fascioliasis on human and animal health. Two novel recombinant Fasciola hepatica proteins, i.e., a ferritin (FhFtn-1) and a tegument-associated protein (FhTP16.5), were used as antigens to develop in-house enzyme-linked immunosorbent assay (ELISA) methods. The assays were optimized and validated using 152 serum samples from humans with a known infection status, including healthy subjects, patients with chronic fascioliasis, and patients with other parasitic diseases. The FhFtn-1 ELISA was shown to be 96.6% sensitive and 95.7% specific; the respective parameters for the FhTP16.5 ELISA were 91.4% and 92.4%. The performances of the FhFtn-1 and FhTP16.5 ELISAs were compared with that of an available commercial test (the DRG test) using a subset of serum samples. Our in-house tests were slightly more sensitive than the DRG test in detecting antibodies against F. hepatica, but the differences were not statistically significant. In conclusion, the present study provides evidence for the potential of the FhFtn-1 and FhTP16.5 ELISAs as diagnostic tools for human fascioliasis, as might be implemented in conjunction with standard assays for large-scale screenings in areas where the disease is endemic and for the detection of occasional cases in clinical laboratories. PMID:24353000

  18. Fasciola hepatica Surface Coat Glycoproteins Contain Mannosylated and Phosphorylated N-glycans and Exhibit Immune Modulatory Properties Independent of the Mannose Receptor.

    Directory of Open Access Journals (Sweden)

    Alessandra Ravidà

    2016-04-01

    Full Text Available Fascioliasis, caused by the liver fluke Fasciola hepatica, is a neglected tropical disease infecting over 1 million individuals annually with 17 million people at risk of infection. Like other helminths, F. hepatica employs mechanisms of immune suppression in order to evade its host immune system. In this study the N-glycosylation of F. hepatica's tegumental coat (FhTeg and its carbohydrate-dependent interactions with bone marrow derived dendritic cells (BMDCs were investigated. Mass spectrometric analysis demonstrated that FhTeg N-glycans comprised mainly of oligomannose and to a lesser extent truncated and complex type glycans, including a phosphorylated subset. The interaction of FhTeg with the mannose receptor (MR was investigated. Binding of FhTeg to MR-transfected CHO cells and BMDCs was blocked when pre-incubated with mannan. We further elucidated the role played by MR in the immunomodulatory mechanism of FhTeg and demonstrated that while FhTeg's binding was significantly reduced in BMDCs generated from MR knockout mice, the absence of MR did not alter FhTeg's ability to induce SOCS3 or suppress cytokine secretion from LPS activated BMDCs. A panel of negatively charged monosaccharides (i.e. GlcNAc-4P, Man-6P and GalNAc-4S were used in an attempt to inhibit the immunoregulatory properties of phosphorylated oligosaccharides. Notably, GalNAc-4S, a known inhibitor of the Cys-domain of MR, efficiently suppressed FhTeg binding to BMDCs and inhibited the expression of suppressor of cytokine signalling (SOCS 3, a negative regulator the TLR and STAT3 pathway. We conclude that F. hepatica contains high levels of mannose residues and phosphorylated glycoproteins that are crucial in modulating its host's immune system, however the role played by MR appears to be limited to the initial binding event suggesting that other C-type lectin receptors are involved in the immunomodulatory mechanism of FhTeg.

  19. Stimulating Neoblast-Like Cell Proliferation in Juvenile Fasciola hepatica Supports Growth and Progression towards the Adult Phenotype In Vitro.

    Directory of Open Access Journals (Sweden)

    Paul McCusker

    2016-09-01

    Full Text Available Fascioliasis (or fasciolosis is a socioeconomically important parasitic disease caused by liver flukes of the genus Fasciola. Flukicide resistance has exposed the need for new drugs and/or a vaccine for liver fluke control. A rapidly improving 'molecular toolbox' for liver fluke encompasses quality genomic/transcriptomic datasets and an RNA interference platform that facilitates functional genomics approaches to drug/vaccine target validation. The exploitation of these resources is undermined by the absence of effective culture/maintenance systems that would support in vitro studies on juvenile fluke development/biology. Here we report markedly improved in vitro maintenance methods for Fasciola hepatica that achieved 65% survival of juvenile fluke after 6 months in standard cell culture medium supplemented with 50% chicken serum. We discovered that this long-term maintenance was dependent upon fluke growth, which was supported by increased proliferation of cells resembling the "neoblast" stem cells described in other flatworms. Growth led to dramatic morphological changes in juveniles, including the development of the digestive tract, reproductive organs and the tegument, towards more adult-like forms. The inhibition of DNA synthesis prevented neoblast-like cell proliferation and inhibited growth/development. Supporting our assertion that we have triggered the development of juveniles towards adult-like fluke, mass spectrometric analyses showed that growing fluke have an excretory/secretory protein profile that is distinct from that of newly-excysted juveniles and more closely resembles that of ex vivo immature and adult fluke. Further, in vitro maintained fluke displayed a transition in their movement from the probing behaviour associated with migrating stage worms to a slower wave-like motility seen in adults. Our ability to stimulate neoblast-like cell proliferation and growth in F. hepatica underpins the first simple platform for their long

  20. Development of two antibody detection enzyme-linked immunosorbent assays for serodiagnosis of human chronic fascioliasis.

    Science.gov (United States)

    Cabán-Hernández, Kimberly; Gaudier, José F; Ruiz-Jiménez, Caleb; Espino, Ana M

    2014-03-01

    Coprological examination based on egg detection in stool samples is currently used as the gold standard for the diagnosis of human fascioliasis. However, this method is not effective during the acute phase of the disease and has poor sensitivity during the chronic phase. Serodiagnosis has become an excellent alternative to coprological examination in efforts to combat the effects of fascioliasis on human and animal health. Two novel recombinant Fasciola hepatica proteins, i.e., a ferritin (FhFtn-1) and a tegument-associated protein (FhTP16.5), were used as antigens to develop in-house enzyme-linked immunosorbent assay (ELISA) methods. The assays were optimized and validated using 152 serum samples from humans with a known infection status, including healthy subjects, patients with chronic fascioliasis, and patients with other parasitic diseases. The FhFtn-1 ELISA was shown to be 96.6% sensitive and 95.7% specific; the respective parameters for the FhTP16.5 ELISA were 91.4% and 92.4%. The performances of the FhFtn-1 and FhTP16.5 ELISAs were compared with that of an available commercial test (the DRG test) using a subset of serum samples. Our in-house tests were slightly more sensitive than the DRG test in detecting antibodies against F. hepatica, but the differences were not statistically significant. In conclusion, the present study provides evidence for the potential of the FhFtn-1 and FhTP16.5 ELISAs as diagnostic tools for human fascioliasis, as might be implemented in conjunction with standard assays for large-scale screenings in areas where the disease is endemic and for the detection of occasional cases in clinical laboratories.

  1. A Herpes Simplex Virus Type 1 Human Asymptomatic CD8+ T-Cell Epitopes-Based Vaccine Protects Against Ocular Herpes in a “Humanized” HLA Transgenic Rabbit Model

    Science.gov (United States)

    Srivastava, Ruchi; Khan, Arif A.; Huang, Jiawei; Nesburn, Anthony B.; Wechsler, Steven L.; BenMohamed, Lbachir

    2015-01-01

    Purpose. A clinical vaccine that protects from ocular herpes simplex virus type 1 (HSV-1) infection and disease still is lacking. In the present study, preclinical vaccine trials of nine asymptomatic (ASYMP) peptides, selected from HSV-1 glycoproteins B (gB), and tegument proteins VP11/12 and VP13/14, were performed in the “humanized” HLA–transgenic rabbit (HLA-Tg rabbit) model of ocular herpes. We recently reported that these peptides are highly recognized by CD8+ T cells from “naturally” protected HSV-1–seropositive healthy ASYMP individuals (who have never had clinical herpes disease). Methods. Mixtures of three ASYMP CD8+ T-cell peptides derived from either HSV-1 gB, VP11/12, or VP13/14 were delivered subcutaneously to different groups of HLA-Tg rabbits (n = 10) in incomplete Freund's adjuvant, twice at 15-day intervals. The frequency and function of HSV-1 epitope-specific CD8+ T cells induced by these peptides and their protective efficacy, in terms of survival, virus replication in the eye, and ocular herpetic disease were assessed after an ocular challenge with HSV-1 (strain McKrae). Results. All mixtures elicited strong and polyfunctional IFN-γ– and TNF-α–producing CD107+CD8+ cytotoxic T cells, associated with a significant reduction in death, ocular herpes infection, and disease (P herpes, and provide a prototype vaccine formulation that may be highly efficacious for preventing ocular herpes in humans. PMID:26098469

  2. Prunasin Hydrolases during Fruit Development in Sweet and Bitter Almonds1[C][W][OA

    Science.gov (United States)

    Sánchez-Pérez, Raquel; Belmonte, Fara Sáez; Borch, Jonas; Dicenta, Federico; Møller, Birger Lindberg; Jørgensen, Kirsten

    2012-01-01

    Amygdalin is a cyanogenic diglucoside and constitutes the bitter component in bitter almond (Prunus dulcis). Amygdalin concentration increases in the course of fruit formation. The monoglucoside prunasin is the precursor of amygdalin. Prunasin may be degraded to hydrogen cyanide, glucose, and benzaldehyde by the action of the β-glucosidase prunasin hydrolase (PH) and mandelonitirile lyase or be glucosylated to form amygdalin. The tissue and cellular localization of PHs was determined during fruit development in two sweet and two bitter almond cultivars using a specific antibody toward PHs. Confocal studies on sections of tegument, nucellus, endosperm, and embryo showed that the localization of the PH proteins is dependent on the stage of fruit development, shifting between apoplast and symplast in opposite patterns in sweet and bitter cultivars. Two different PH genes, Ph691 and Ph692, have been identified in a sweet and a bitter almond cultivar. Both cDNAs are 86% identical on the nucleotide level, and their encoded proteins are 79% identical to each other. In addition, Ph691 and Ph692 display 92% and 86% nucleotide identity to Ph1 from black cherry (Prunus serotina). Both proteins were predicted to contain an amino-terminal signal peptide, with the size of 26 amino acid residues for PH691 and 22 residues for PH692. The PH activity and the localization of the respective proteins in vivo differ between cultivars. This implies that there might be different concentrations of prunasin available in the seed for amygdalin synthesis and that these differences may determine whether the mature almond develops into bitter or sweet. PMID:22353576

  3. Some Diagnosis and Therapy Techniques in Rainbow Trout (Oncorhynchus Mykiss Furunculosis, in Intensive System Bred

    Directory of Open Access Journals (Sweden)

    Octavian Negrea

    2010-10-01

    Full Text Available Clinical and paraclinical investigations (morpho-pathologic, bacterioscopic, bacteriologic and sensitiveness test toantibodies done on 20 fish (rainbow trout, dead or in agony state, taken from a furunculosis pesthole in Salmonidaeand proceeded from an intensive exploitation unit of trout, in Cluj county, put in evidence the following aspects :Toclinical exam, in breeding basins, it appears individuals (5 % which have presented locomotor troubles, listless anduntidy swimming in water surface, lateral swimming anorexia, and on tegument it is observed diffuse hemorrhagiczones, lost of fish scale and necrosis of fins. It appears also secondary infections with fungi of Saprolegnia genusunder the form of a white- dirty, downy film. From the 20 trut corpses were isolated 2 bacterial strains, from anteriorkidney and from raclage in fin basis, with congestive lesions. Bacteria isolated are developing only after anincubation at 20 – 250 C and does not grow up to 370 C, and to microscopic exam directly from sauce, the mobility isabsent Classic biochemical tests put in evidence the following positive biochemic properties: cataloze, oxidaze,indolal and the presence of hemolytic activity. Biochemical properties testing on API20E gallery also puts inevidence properties as : positive gelatinousis and sucraze fermentation. Based on bacterial strains developmentisolated only to 20 - 250C and the above mentioned biochemic properties, respectively the absence of pigment,bacterial strains isolated are appointed in Aeromonas genus, Aeromonas salmonicida species, achromogenessubspecies. By sensitiveness test in different medicine substances, using antibiogram technique, bacterial strainsisolated from pesthole are sensitive in a decreasing order to: nalidixic acid, oxitetracycline, florfenicol andeurofloxacin and resistant to ampicyline, amoxiclav and colistin.

  4. Serotonin Signaling in Schistosoma mansoni: A Serotonin–Activated G Protein-Coupled Receptor Controls Parasite Movement

    Science.gov (United States)

    Rashid, Mohammed; Ribeiro, Paula

    2014-01-01

    Serotonin is an important neuroactive substance in all the parasitic helminths. In Schistosoma mansoni, serotonin is strongly myoexcitatory; it potentiates contraction of the body wall muscles and stimulates motor activity. This is considered to be a critical mechanism of motor control in the parasite, but the mode of action of serotonin is poorly understood. Here we provide the first molecular evidence of a functional serotonin receptor (Sm5HTR) in S. mansoni. The schistosome receptor belongs to the G protein-coupled receptor (GPCR) superfamily and is distantly related to serotonergic type 7 (5HT7) receptors from other species. Functional expression studies in transfected HEK 293 cells showed that Sm5HTR is a specific serotonin receptor and it signals through an increase in intracellular cAMP, consistent with a 5HT7 signaling mechanism. Immunolocalization studies with a specific anti-Sm5HTR antibody revealed that the receptor is abundantly distributed in the worm's nervous system, including the cerebral ganglia and main nerve cords of the central nervous system and the peripheral innervation of the body wall muscles and tegument. RNA interference (RNAi) was performed both in schistosomulae and adult worms to test whether the receptor is required for parasite motility. The RNAi-suppressed adults and larvae were markedly hypoactive compared to the corresponding controls and they were also resistant to exogenous serotonin treatment. These results show that Sm5HTR is at least one of the receptors responsible for the motor effects of serotonin in S. mansoni. The fact that Sm5HTR is expressed in nerve tissue further suggests that serotonin stimulates movement via this receptor by modulating neuronal output to the musculature. Together, the evidence identifies Sm5HTR as an important neuronal protein and a key component of the motor control apparatus in S. mansoni. PMID:24453972

  5. SOMATIC EMBRYOGENESIS AND MORPHOANATOMY OF Ocotea porosa SOMATIC EMBRYOS

    Directory of Open Access Journals (Sweden)

    Luciana Luiza Pelegrini

    2013-12-01

    Full Text Available http://dx.doi.org/10.5902/1980509812343Ocotea porosa seeds have strong tegument dormancy, recalcitrant behavior, low and irregular germinationand that makes its natural propagation difficult. The aim of this study was to establish a protocol ofregeneration of Ocotea porosa from somatic embryogenesis. Immature embryonic axes were inoculatedon WPM culture medium supplemented with 2.4-D (200 μM combined or not with hydrolyzed casein orglutamine (0.5 or 1 g l-1, during 90 days. The repetitive embryogenesis was induced on medium with 2.4-D(22.62 μM combined with 2-iP (2.46 μM followed by transfer to culture medium with hydrolyzed caseinor glutamine (1 g l-1 during 90 days. The maturation of somatic embryos was tested in culture mediumcontaining NAA (0.5 μM and 2-iP (5; 10 and 20 μM. The highest percentage of somatic embryos induction(8.3% was observed in WPM culture medium containing 200 μM 2.4-D and 1 g L-1 hydrolyzed casein andthe development of somatic embryos occurred indirectly. Repetitive somatic embryogenesis was promotedin WPM medium containing hydrolyzed casein or glutamine. However, the culture medium containinghydrolyzed casein promoted the maintenance of embryogenic capacity for more than two years. Duringthe maturity phase, there was a low progression of globular embryos to cordiform and torpedo stages.The different ontogenetic stages of somatic embryos of Ocotea porosa were characterized by histologicalstudies.

  6. Developmental gene expression profiles of the human pathogen Schistosoma japonicum

    Directory of Open Access Journals (Sweden)

    McManus Donald P

    2009-03-01

    Full Text Available Abstract Background The schistosome blood flukes are complex trematodes and cause a chronic parasitic disease of significant public health importance worldwide, schistosomiasis. Their life cycle is characterised by distinct parasitic and free-living phases involving mammalian and snail hosts and freshwater. Microarray analysis was used to profile developmental gene expression in the Asian species, Schistosoma japonicum. Total RNAs were isolated from the three distinct environmental phases of the lifecycle – aquatic/snail (eggs, miracidia, sporocysts, cercariae, juvenile (lung schistosomula and paired but pre-egg laying adults and adult (paired, mature males and egg-producing females, both examined separately. Advanced analyses including ANOVA, principal component analysis, and hierarchal clustering provided a global synopsis of gene expression relationships among the different developmental stages of the schistosome parasite. Results Gene expression profiles were linked to the major environmental settings through which the developmental stages of the fluke have to adapt during the course of its life cycle. Gene ontologies of the differentially expressed genes revealed a wide range of functions and processes. In addition, stage-specific, differentially expressed genes were identified that were involved in numerous biological pathways and functions including calcium signalling, sphingolipid metabolism and parasite defence. Conclusion The findings provide a comprehensive database of gene expression in an important human pathogen, including transcriptional changes in genes involved in evasion of the host immune response, nutrient acquisition, energy production, calcium signalling, sphingolipid metabolism, egg production and tegumental function during development. This resource should help facilitate the identification and prioritization of new anti-schistosome drug and vaccine targets for the control of schistosomiasis.

  7. Morphology and histochemistry of the aesthetasc-associated epidermal glands in terrestrial hermit crabs of the genus Coenobita (Decapoda: Paguroidea.

    Directory of Open Access Journals (Sweden)

    Oksana Tuchina

    Full Text Available Crustaceans have successfully adapted to a variety of environments including fresh- and saltwater as well as land. Transition from an aquatic to a terrestrial lifestyle required adaptations of the sensory equipment of an animal, particularly in olfaction, where the stimulus itself changes from hydrophilic to mainly hydrophobic, air-borne molecules. Hermit crabs Coenobita spp. (Anomura, Coenobitidae have adapted to a fully terrestrial lifestyle as adults and have been shown to rely on olfaction in order to detect distant food items. We observed that the specialized olfactory sensilla in Coenobita, named aesthetascs, are immersed in a layer of mucous-like substance. We hypothesized that the mucous is produced by antennal glands and affects functioning of the aesthetascs. Using various microscopic and histochemical techniques we proved that the mucous is produced by aesthetasc-associated epidermal glands, which we consider to be modified rosette-type aesthetasc tegumental glands known from aquatic decapods. These epidermal glands in Coenobita are multicellular exocrine organs of the recto-canal type with tubulo-acinar arrangement of the secretory cells. Two distinct populations of secretory cells were clearly distinguishable with light and electron microscopy. At least part of the secretory cells contains specific enzymes, CUB-serine proteases, which are likely to be secreted on the surface of the aesthetasc pad and take part in antimicrobial defense. Proteomic analysis of the glandular tissue corroborates the idea that the secretions of the aesthetasc-associated epidermal glands are involved in immune responses. We propose that the mucous covering the aesthetascs in Coenobita takes part in antimicrobial defense and at the same time provides the moisture essential for odor perception in terrestrial hermit crabs. We conclude that the morphological modifications of the aesthetasc-associated epidermal glands as well as the functional characteristics

  8. Characterization of a Gene Family Encoding SEA (Sea-urchin Sperm Protein, Enterokinase and Agrin)-Domain Proteins with Lectin-Like and Heme-Binding Properties from Schistosoma japonicum

    Science.gov (United States)

    Mbanefo, Evaristus Chibunna; Kikuchi, Mihoko; Huy, Nguyen Tien; Shuaibu, Mohammed Nasir; Cherif, Mahamoud Sama; Yu, Chuanxin; Wakao, Masahiro; Suda, Yasuo; Hirayama, Kenji

    2014-01-01

    Background We previously identified a novel gene family dispersed in the genome of Schistosoma japonicum by retrotransposon-mediated gene duplication mechanism. Although many transcripts were identified, no homolog was readily identifiable from sequence information. Methodology/Principal Findings Here, we utilized structural homology modeling and biochemical methods to identify remote homologs, and characterized the gene products as SEA (sea-urchin sperm protein, enterokinase and agrin)-domain containing proteins. A common extracellular domain in this family was structurally similar to SEA-domain. SEA-domain is primarily a structural domain, known to assist or regulate binding to glycans. Recombinant proteins from three members of this gene family specifically interacted with glycosaminoglycans with high affinity, with potential implication in ligand acquisition and immune evasion. Similar approach was used to identify a heme-binding site on the SEA-domain. The heme-binding mode showed heme molecule inserted into a hydrophobic pocket, with heme iron putatively coordinated to two histidine axial ligands. Heme-binding properties were confirmed using biochemical assays and UV-visible absorption spectroscopy, which showed high affinity heme-binding (K D = 1.605×10−6 M) and cognate spectroscopic attributes of hexa-coordinated heme iron. The native proteins were oligomers, antigenic, and are localized on adult worm teguments and gastrodermis; major host-parasite interfaces and site for heme detoxification and acquisition. Conclusions The results suggest potential role, at least in the nucleation step of heme crystallization (hemozoin formation), and as receptors for heme uptake. Survival strategies exploited by parasites, including heme homeostasis mechanism in hemoparasites, are paramount for successful parasitism. Thus, assessing prospects for application in disease intervention is warranted. PMID:24416467

  9. Exocrine glands of Lepeophtheirus salmonis (Copepoda: Caligidae): Distribution, developmental appearance, and site of secretion.

    Science.gov (United States)

    Øvergård, Aina-Cathrine; Hamre, Lars A; Harasimczuk, Ewa; Dalvin, Sussie; Nilsen, Frank; Grotmol, Sindre

    2016-12-01

    Exocrine glands of blood-feeding parasitic copepods are believed to be important in host immune response modulation and inhibition of host blood coagulation, but also in the production of substances for integument lubrication and antifouling. In this study, we aimed to characterize the distribution of different types of salmon louse (Lepeophtheirus salmonis) exocrine glands and their site of secretion. The developmental appearance of each gland type was mapped and genes specifically expressed by glands were identified. Three types of tegumental (teg 1-3) glands and one labial gland type were found. The first glands to appear during development were teg 1 and teg 2 glands. They have ducts extending both dorsally and ventrally suggested to be important in lubricating the integument. Teg 1 glands were found to express two astacin metallopeptidases and a gene with fibronectin II domains, while teg 2 glands express a heme peroxidase. The labial glands were first identified in planktonic copepodids, with reservoirs that allows for storage of glandular products. The last gland type to appear during development was named teg 3 and was not seen before the preadult I stage when the lice become more virulent. Teg 3 glands have ducts ending ventrally at the host-parasite contact area, and may secrete substances important for the salmon lice virulence. Salmon lice teg 3 and labial glands are thus likely to be especially important in the host-parasite interaction. Proteins secreted from the salmon louse glands to its salmonid host skin or blood represents a potential interface where the host immune system can meet and elicit effective responses to sea lice antigens. The present study thus represents a fundamental basis for further functional studies and identification of possible vaccine candidates. J. Morphol. 277:1616-1630, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  10. The shifting zoom: new possibilities for inverse scattering on electrically large domains

    Science.gov (United States)

    Persico, Raffaele; Ludeno, Giovanni; Soldovieri, Francesco; De Coster, Alberic; Lambot, Sebastien

    2017-04-01

    . Atzeni, R. Persico, F. Soldovieri, Advanced Processing Techniques for Step-frequency Continuous-Wave Penetrating Radar: the Case Study of "Palazzo Vecchio" Walls (Firenze, Italy), Research on Nondestructive Evaluation, vol. 17, pp. 71-83, 2006. [2] N. Masini, R. Persico, E. Rizzo, A. Calia, M. T. Giannotta, G. Quarta, A. Pagliuca, "Integrated Techniques for Analysis and Monitoring of Historical Monuments: the case of S.Giovanni al Sepolcro in Brindisi (Southern Italy)." Near Surface Geophysics, vol. 8 (5), pp. 423-432, 2010. [3] E. Pettinelli, A. Di Matteo, E. Mattei, L. Crocco, F. Soldovieri, J. D. Redman, and A. P. Annan, "GPR response from buried pipes: Measurement on field site and tomographic reconstructions", IEEE Transactions on Geoscience and Remote Sensing, vol. 47, n. 8, 2639-2645, Aug. 2009. [4] O. Lopera, E. C. Slob, N. Milisavljevic and S. Lambot, "Filtering soil surface and antenna effects from GPR data to enhance landmine detection", IEEE Transactions on Geoscience and Remote Sensing, vol. 45, n. 3, pp.707-717, 2007. [5] R. Persico, "Introduction to Ground Penetrating Radar: Inverse Scattering and Data Processing". Wiley, 2014. [6] R. Persico, J. Sala, "The problem of the investigation domain subdivision in 2D linear inversions for large scale GPR data", IEEE Geoscience and Remote Sensing Letters, vol. 11, n. 7, pp. 1215-1219, doi 10.1109/LGRS.2013.2290008, July 2014. [7] R. Persico, F. Soldovieri, S. Lambot, Shifting zoom in 2D linear inversions performed on GPR data gathered along an electrically large investigation domain, Proc. 16th International Conference on Ground Penetrating Radar GPR2016, Honk-Kong, June 13-16, 2016

  11. Revised stratigraphy and reinterpretation of the Miocene Pohang basinfill, SE Korea: sequence development in response to tectonism and eustasy in a back-arc basin margin

    Science.gov (United States)

    Sohn, Y. K.; Rhee, C. W.; Shon, H.

    2001-09-01

    -level fluctuations. We suggest that the four alternations of conglomerates (lowstand systems) and hemipelagic mudstones (condensed intervals) resulted most probably from the 3rd-order glacioeustatic cycles during the middle Miocene. This finding implies that the signatures of global sea-level fluctuations can be deciphered from a tectonically active sedimentary basin if the timing of regional tectonic development is well constrained, and the global sea-level chart of Haq et al. ( Haq, B.W., Hardenbol, J., Vail, P.R., 1987, Chronology of fluctuating sea levels since the Triassic. Science 235, 1156-1167; Haq, B.U., Hardenbol, J., Vail, P.R., 1988. Mesozoic and Cenozoic chronostratigraphy and eustatic cycles. In: Wilgus, C.K., Hastings, B.S., Posamentier, H., Van Wagoner, J., Ross, C.A., Kendall, C.G.S.C. (Eds.), Sea-Level Changes: an Integrated Approach: Soc. Econ. Paleont. Mineral. Spec. Publ. 42, pp. 71-108) may serve as a guide to basinfill interpretation even in tectonically active sedimentary basins.

  12. Ground Penetrating Radar employment for searching ancient cisterns.

    Science.gov (United States)

    Semeraro, Grazia; Notario, Corrado; Persico, Raffaele

    2017-04-01

    , M. Noviello, M. Ciminale, R. Persico, Integration of multisensor data: an experiment in the archaeological park of Egnazia (Apulia, Southern Italy), Near Surface Geophysics, Vol. 13, n. 6, pp. 613-621, 2015 4) G. Leucci, N. Masini, R. Persico, F. Soldovieri." GPR and sonic tomography for structural restoration : the case of the Cathedral of Tricarico", Journal of Geophysics and Engineering, vol. 8, pp. S76-S92, Aug. 2011. 5) M. Pieraccini, L. Noferini, D. Mecatti, C. Atzeni, R. Persico, F. Soldovieri, Advanced Processing Techniques for Step-frequency Continuous-Wave Penetrating Radar: the Case Study of "Palazzo Vecchio" Walls (Firenze, Italy), Research on Nondestructive Evaluation, vol. 17, pp. 71-83, 2006. 6) N. Masini, R. Persico, E. Rizzo, A. Calia, M.T. Giannotta, G. Quarta, A. Pagliuca, "Integrated Techniques for Analysis and Monitoring of Historical Monuments: the case of S.Giovanni al Sepolcro in Brindisi (Southern Italy)." Near Surface Geophysics, vol. 8, n. 5, pp. 423-432, 2010.

  13. Polypyridylruthenium(II complexes exert anti-schistosome activity and inhibit parasite acetylcholinesterases.

    Directory of Open Access Journals (Sweden)

    Madhu K Sundaraneedi

    2017-12-01

    Full Text Available Schistosomiasis affects over 200 million people and there are concerns whether the current chemotherapeutic control strategy (periodic mass drug administration with praziquantel (PZQ-the only licenced anti-schistosome compound is sustainable, necessitating the development of new drugs.We investigated the anti-schistosome efficacy of polypyridylruthenium(II complexes and showed they were active against all intra-mammalian stages of S. mansoni. Two compounds, Rubb12-tri and Rubb7-tnl, which were among the most potent in their ability to kill schistosomula and adult worms and inhibit egg hatching in vitro, were assessed for their efficacy in a mouse model of schistosomiasis using 5 consecutive daily i.v. doses of 2 mg/kg (Rubb12-tri and 10 mg/kg (Rubb7-tnl. Mice treated with Rubb12-tri showed an average 42% reduction (P = 0.009, over two independent trials, in adult worm burden. Liver egg burdens were not significantly decreased in either drug-treated group but ova from both of these groups showed significant decreases in hatching ability (Rubb12-tri-68%, Rubb7-tnl-56% and were significantly morphologically altered (Rubb12-tri-62% abnormal, Rubb7-tnl-35% abnormal. We hypothesize that the drugs exerted their activity, at least partially, through inhibition of both neuronal and tegumental acetylcholinesterases (AChEs, as worms treated in vitro showed significant decreases in activity of these enzymes. Further, treated parasites exhibited a significantly decreased ability to uptake glucose, significantly depleted glycogen stores and withered tubercules (a site of glycogen storage, implying drug-mediated interference in this nutrient acquisition pathway.Our data provide compelling evidence that ruthenium complexes are effective against all intra-mammalian stages of schistosomes, including schistosomula (refractory to PZQ and eggs (agents of disease transmissibility. Further, the results of this study suggest that schistosome AChE is a target of

  14. Antiparasitic activity of menadione (vitamin K3) against Schistosoma mansoni in BABL/c mice.

    Science.gov (United States)

    Kapadia, Govind J; Soares, Ingrid A O; Rao, G Subba; Badoco, Fernanda R; Furtado, Ricardo A; Correa, Mariana B; Tavares, Denise C; Cunha, Wilson R; Magalhães, Lizandra G

    2017-03-01

    Schistosomiasis is one of the neglected tropical diseases affecting nearly quarter of a billion people in economically challenged tropical and subtropical countries of the world. Praziquantel (PZQ) is the only drug currently available to treat this parasitic disease in spite being ineffective against juvenile worms and concerns about developing resistance to treat reinfections. Our earlier in vitro viability studies demonstrated significant antiparasitic activity of menadione (MEN) (vitamin K 3 ) against Schistosoma mansoni adult worms. To gain insight into plausible mechanism of antischistosomal activity of MEN, its effect on superoxide anion levels in adult worms were studied in vitro which showed significant increases in both female and male worms. Further confirmation of the deleterious morphological changes in their teguments and organelles were obtained by ultrastructural analysis. Genotoxic and cytotoxic studies in male Swiss mice indicated that MEN was well tolerated at the oral dose of 500mg/kg using the criteria of MNPCE frequency and PCE/RBC ratio in the bone marrow of infected animals. The in vivo antiparasitic activity of MEN was conducted in female BALB/c mice infected with S. mansoni and significant reductions (P<0.001) in total worm burden were observed at single oral doses of 40 and 400mg/kg (48.57 and 61.90%, respectively). Additionally, MEN significantly reduced (P<0.001) the number of eggs in the liver of infected mice by 53.57 and 58.76%, respectively. Similarly, histological analysis of the livers showed a significant reduction (P<0.001) in the diameter of the granulomas. Since MEN is already in use globally as an over-the-counter drug for a variety of common ailments and a dietary supplement with a safety record in par with similar products when used in recommended doses, the above antiparasitic results which compare reasonably well with PZQ, make a compelling case for considering MEN to treat S. mansoni infection in humans. Copyright © 2016

  15. First insight into CD59-like molecules of adult Fasciola hepatica.

    Science.gov (United States)

    Shi, Yunliang; Toet, Hayley; Rathinasamy, Vignesh; Young, Neil D; Gasser, Robin B; Beddoe, Travis; Huang, Weiyi; Spithill, Terry W

    2014-09-01

    The present study focussed on investigating CD59-like molecules of Fasciola hepatica. A cDNA encoding a CD59-like protein (termed FhCD59-1) identified previously in the membrane fraction of the F. hepatica tegument was isolated. This homologue was shown to encode a predicted open reading frame (ORF) of 122 amino acids (aa) orthologous to human CD59 with a 25 aa signal peptide, a mature protein containing 10 cysteines and a conserved CD59/Ly-6 family motif "CCXXXXCN". An analysis of cDNAs from two different adult specimens of F. hepatica revealed seven variable types of FhCD59-1 sequences, designated FhCD59-1.1 to FhCD59-1.7, which had 94.3-99.7% amino acid sequence identity upon pairwise comparison. Molecular modeling of FhCD59-1.1 with human CD59 confirmed the presence of the three-finger protein domain found in the CD59 family and predicted three disulphide bonds in the F. hepatica sequence. The interrogation of F. hepatica databases identified two additional sequences, designated FhCD59-2 and FhCD59-3, which had only 23.4-29.5% amino acid identity to FhCD59-1.1. Orthologues of the inferred CD59 protein sequences of F. hepatica were also identified in other flatworms, including Fasciola gigantica, Fascioloides magna, Schistosoma haematobium, Schistosoma japonicum, Schistosoma mansoni, Clonorchis sinensis, Opisthorchis viverrini, Taenia solium, Echinococcus granulosus and the free living Schmidtea mediterannea. The results revealed a considerable degree of sequence complexity in the CD59-like sequence families in F. hepatica and flatworms. Phylogenetic analysis of CD59-like aa sequences from F. hepatica and flatworms showed that FhCD59-2 clustered with the known surface-associated protein SmCD59-2 of S. mansoni. Relatively well-supported clades specific to schistosomes, fasciolids and opisthorchiids were identified. The qPCR analysis of gene transcription showed that the relative expression of these 3 FhCD59-like sequences varied by 11-47-fold during fluke

  16. DNA Vaccine Encoding the Chimeric Form of Schistosoma mansoni Sm-TSP2 and Sm29 Confers Partial Protection against Challenge Infection

    Science.gov (United States)

    Gonçalves de Assis, Natan Raimundo; Batistoni de Morais, Suellen; Figueiredo, Bárbara Castro Pimentel; Ricci, Natasha Delaqua; de Almeida, Leonardo Augusto; da Silva Pinheiro, Carina; Martins, Vicente de Paulo; Oliveira, Sergio Costa

    2015-01-01

    Schistosomiasis is an important parasitic disease worldwide that affects more than 207 million people in 76 countries and causes approximately 250,000 deaths per year. The best long-term strategy to control schistosomiasis is through immunization combined with drug treatment. Due to the ability of DNA vaccines to generate humoral and cellular immune responses, such vaccines are considered a promising approach against schistosomiasis. Sm29 and tetraspanin-2 (Sm-TSP2) are two proteins that are located in the S. mansoni tegument of adult worms and schistosomula and induce high levels of protection through recombinant protein immunization. In this study, we transfected BHK-21 cells with plasmids encoding Sm29, Sm-TSP2 or a chimera containing both genes. Using RT-PCR analysis and western blot, we confirmed that the DNA vaccine constructs were transcribed and translated, respectively, in BHK-21 cells. After immunization of mice, we evaluated the reduction in worm burden. We observed worm burden reductions of 17-22%, 22%, 31-32% and 24-32% in animals immunized with the pUMVC3/Sm29, pUMVC3/SmTSP-2, pUMVC3/Chimera and pUMVC3/Sm29 + pUMVC3/SmTSP-2 plasmids, respectively. We evaluated the humoral response elicited by DNA vaccines, and animals immunized with pUMVC3/Sm29 and pUMVC3/Sm29 + pUMVC3/SmTSP-2 showed higher titers of anti-Sm29 antibodies. The cytokine profile produced by the spleen cells of immunized mice was then evaluated. We observed higher production of Th1 cytokines, such as TNF-α and IFN-γ, in vaccinated mice and no significant production of IL-4 and IL-5. The DNA vaccines tested in this study showed the ability to generate a protective immune response against schistosomiasis, probably through the production of Th1 cytokines. However, future strategies aiming to optimize the protective response induced by a chimeric DNA construct need to be developed. PMID:25942636

  17. Cloning and characterisation of Schistosoma japonicum insulin receptors.

    Directory of Open Access Journals (Sweden)

    Hong You

    2010-03-01

    Full Text Available Schistosomes depend for growth and development on host hormonal signals, which may include the insulin signalling pathway. We cloned and assessed the function of two insulin receptors from Schistosoma japonicum in order to shed light on their role in schistosome biology.We isolated, from S. japonicum, insulin receptors 1 (SjIR-1 and 2 (SjIR-2 sharing close sequence identity to their S. mansoni homologues (SmIR-1 and SmIR-2. SjIR-1 is located on the tegument basal membrane and the internal epithelium of adult worms, whereas SjIR-2 is located in the parenchyma of males and the vitelline tissue of females. Phylogenetic analysis showed that SjIR-2 and SmIR-2 are close to Echinococcus multilocularis insulin receptor (EmIR, suggesting that SjIR-2, SmIR-2 and EmIR share similar roles in growth and development in the three taxa. Structure homology modelling recovered the conserved structure between the SjIRs and Homo sapiens IR (HIR implying a common predicted binding mechanism in the ligand domain and the same downstream signal transduction processing in the tyrosine kinase domain as in HIR. Two-hybrid analysis was used to confirm that the ligand domains of SjIR-1 and SjIR-2 contain the insulin binding site. Incubation of adult worms in vitro, both with a specific insulin receptor inhibitor and anti-SjIRs antibodies, resulted in a significant decrease in worm glucose levels, suggesting again the same function for SjIRs in regulating glucose uptake as described for mammalian cells.Adult worms of S. japonicum possess insulin receptors that can specifically bind to insulin, indicating that the parasite can utilize host insulin for development and growth by sharing the same pathway as mammalian cells in regulating glucose uptake. A complete understanding of the role of SjIRs in the biology of S. japonicum may result in their use as new targets for drug and vaccine development against schistosomiasis.

  18. A Survey of Mucilaginous Testa in Chamaesyce

    Directory of Open Access Journals (Sweden)

    Jordan, M. S.

    1992-12-01

    cèl-lules superficials la que secreta mucilags. Les cèl-lules secretores de mucílags i les macroesclereides de la testa madura s' interpreten com desenvolupades a partir de les capes epidèrmiques dels teguments extern i intern, respectivament. El mucilag de les granes possiblement juga un paper en la hidratació i/o dispersió de les granes.

  19. The Translesion Polymerase Pol η Is Required for Efficient Epstein-Barr Virus Infectivity and Is Regulated by the Viral Deubiquitinating Enzyme BPLF1.

    Science.gov (United States)

    Dyson, Ossie F; Pagano, Joseph S; Whitehurst, Christopher B

    2017-10-01

    Epstein-Barr virus (EBV) infection and lytic replication are known to induce a cellular DNA damage response. We previously showed that the virally encoded BPLF1 protein interacts with and regulates several members of the translesion synthesis (TLS) pathway, a DNA damage tolerance pathway, and that these cellular factors enhance viral infectivity. BPLF1 is a late lytic cycle gene, but the protein is also packaged in the viral tegument, indicating that BPLF1 may function both early and late during infection. The BPLF1 protein expresses deubiquitinating activity that is strictly conserved across the Herpesviridae ; mutation of the active site cysteine results in a loss of enzymatic activity. Infection with an EBV BPLF1 knockout virus results in decreased EBV infectivity. Polymerase eta (Pol η), a specialized DNA repair polymerase, functions in TLS and allows for DNA replication complexes to bypass lesions in DNA. Here we report that BPLF1 interacts with Pol η and that Pol η protein levels are increased in the presence of functional BPLF1. BPLF1 promotes a nuclear relocalization of Pol η molecules which are focus-like in appearance, consistent with the localization observed when Pol η is recruited to sites of DNA damage. Knockdown of Pol η resulted in decreased production of infectious virus, and further, Pol η was found to bind to EBV DNA, suggesting that it may allow for bypass of damaged viral DNA during its replication. The results suggest a mechanism by which EBV recruits cellular repair factors, such as Pol η, to sites of viral DNA damage via BPLF1, thereby allowing for efficient viral DNA replication. IMPORTANCE Epstein-Barr virus is the causative agent of infectious mononucleosis and infects approximately 90% of the world's population. It causes lymphomas in individuals with acquired and innate immune disorders and is strongly associated with Hodgkin's lymphoma, Burkitt's lymphoma, diffuse large B-cell lymphomas, nasopharyngeal carcinoma (NPC), and

  20. Epstein-Barr Virus BKRF4 Gene Product Is Required for Efficient Progeny Production.

    Science.gov (United States)

    Masud, H M Abdullah Al; Watanabe, Takahiro; Yoshida, Masahiro; Sato, Yoshitaka; Goshima, Fumi; Kimura, Hiroshi; Murata, Takayuki

    2017-12-01

    Epstein-Barr virus (EBV), a member of human gammaherpesvirus, infects mainly B cells. EBV has two alternative life cycles, latent and lytic, and is reactivated occasionally from the latent stage to the lytic cycle. To combat EBV-associated disorders, understanding the molecular mechanisms of the EBV lytic replication cycle is also important. Here, we focused on an EBV lytic gene, BKRF4. Using our anti-BKRF4 antibody, we revealed that the BKRF4 gene product is expressed during the lytic cycle with late kinetics. To characterize the role of BKRF4, we constructed BKRF4-knockout mutants using the bacterial artificial chromosome (BAC) and CRISPR/Cas9 systems. Although disruption of the BKRF4 gene had almost no effect on viral protein expression and DNA synthesis, it significantly decreased progeny virion levels in HEK293 and Akata cells. Furthermore, we show that BKRF4 is involved not only in production of progeny virions but also in increasing the infectivity of the virus particles. Immunoprecipitation assays revealed that BKRF4 interacted with a virion protein, BGLF2. We showed that the C-terminal region of BKRF4 was critical for this interaction and for efficient progeny production. Immunofluorescence analysis revealed that BKRF4 partially colocalized with BGLF2 in the nucleus and perinuclear region. Finally, we showed that BKRF4 is a phosphorylated, possible tegument protein and that the EBV protein kinase BGLF4 may be important for this phosphorylation. Taken together, our data suggest that BKRF4 is involved in the production of infectious virions. IMPORTANCE Although the latent genes of EBV have been studied extensively, the lytic genes are less well characterized. This study focused on one such lytic gene, BKRF4, which is conserved only among gammaherpesviruses (ORF45 of Kaposi's sarcoma-associated herpesvirus or murine herpesvirus 68). After preparing the BKRF4 knockout virus using B95-8 EBV-BAC, we demonstrated that the BKRF4 gene was involved in infectious

  1. DNA Vaccine Encoding the Chimeric Form of Schistosoma mansoni Sm-TSP2 and Sm29 Confers Partial Protection against Challenge Infection.

    Directory of Open Access Journals (Sweden)

    Natan Raimundo Gonçalves de Assis

    Full Text Available Schistosomiasis is an important parasitic disease worldwide that affects more than 207 million people in 76 countries and causes approximately 250,000 deaths per year. The best long-term strategy to control schistosomiasis is through immunization combined with drug treatment. Due to the ability of DNA vaccines to generate humoral and cellular immune responses, such vaccines are considered a promising approach against schistosomiasis. Sm29 and tetraspanin-2 (Sm-TSP2 are two proteins that are located in the S. mansoni tegument of adult worms and schistosomula and induce high levels of protection through recombinant protein immunization. In this study, we transfected BHK-21 cells with plasmids encoding Sm29, Sm-TSP2 or a chimera containing both genes. Using RT-PCR analysis and western blot, we confirmed that the DNA vaccine constructs were transcribed and translated, respectively, in BHK-21 cells. After immunization of mice, we evaluated the reduction in worm burden. We observed worm burden reductions of 17-22%, 22%, 31-32% and 24-32% in animals immunized with the pUMVC3/Sm29, pUMVC3/SmTSP-2, pUMVC3/Chimera and pUMVC3/Sm29 + pUMVC3/SmTSP-2 plasmids, respectively. We evaluated the humoral response elicited by DNA vaccines, and animals immunized with pUMVC3/Sm29 and pUMVC3/Sm29 + pUMVC3/SmTSP-2 showed higher titers of anti-Sm29 antibodies. The cytokine profile produced by the spleen cells of immunized mice was then evaluated. We observed higher production of Th1 cytokines, such as TNF-α and IFN-γ, in vaccinated mice and no significant production of IL-4 and IL-5. The DNA vaccines tested in this study showed the ability to generate a protective immune response against schistosomiasis, probably through the production of Th1 cytokines. However, future strategies aiming to optimize the protective response induced by a chimeric DNA construct need to be developed.

  2. The Schistosoma mansoni phylome: using evolutionary genomics to gain insight into a parasite’s biology

    Directory of Open Access Journals (Sweden)

    Silva Larissa

    2012-11-01

    Full Text Available Abstract Background Schistosoma mansoni is one of the causative agents of schistosomiasis, a neglected tropical disease that affects about 237 million people worldwide. Despite recent efforts, we still lack a general understanding of the relevant host-parasite interactions, and the possible treatments are limited by the emergence of resistant strains and the absence of a vaccine. The S. mansoni genome was completely sequenced and still under continuous annotation. Nevertheless, more than 45% of the encoded proteins remain without experimental characterization or even functional prediction. To improve our knowledge regarding the biology of this parasite, we conducted a proteome-wide evolutionary analysis to provide a broad view of the S. mansoni’s proteome evolution and to improve its functional annotation. Results Using a phylogenomic approach, we reconstructed the S. mansoni phylome, which comprises the evolutionary histories of all parasite proteins and their homologs across 12 other organisms. The analysis of a total of 7,964 phylogenies allowed a deeper understanding of genomic complexity and evolutionary adaptations to a parasitic lifestyle. In particular, the identification of lineage-specific gene duplications pointed to the diversification of several protein families that are relevant for host-parasite interaction, including proteases, tetraspanins, fucosyltransferases, venom allergen-like proteins, and tegumental-allergen-like proteins. In addition to the evolutionary knowledge, the phylome data enabled us to automatically re-annotate 3,451 proteins through a phylogenetic-based approach rather than solely sequence similarity searches. To allow further exploitation of this valuable data, all information has been made available at PhylomeDB (http://www.phylomedb.org. Conclusions In this study, we used an evolutionary approach to assess S. mansoni parasite biology, improve genome/proteome functional annotation, and provide insights into

  3. Consideraciones cardiovasculares del síndrome de Marfán en edades pediátricas Cardiovascular considerations about Marfan's syndrome at pediatric ages

    Directory of Open Access Journals (Sweden)

    Giselle Serrano Ricardo

    2012-06-01

    Full Text Available El síndrome de Marfán es una enfermedad hereditaria del tejido conectivo, que se describe en niños y en adultos, causada por una mutación en el gen que codifica la glicoproteína fibrilina tipo 1. Afecta múltiples órganos y sistemas, fundamentalmente cardiovascular, esquelético, oftalmológico, piel y tegumentos. Se presenta una revisión de los aspectos más actuales del diagnóstico, y la atención multidisciplinaria para lograr una reducción de la morbilidad y mortalidad en los pacientes pediátricos. Se concluye que el uso precoz de betabloqueadores e inhibidores del receptor AT-1 de la angiotensina II (losartán, constituyen actualmente los pilares fundamentales de la terapéutica farmacológica, pues disminuyen la frecuencia de complicaciones cardiovasculares, las cuales determinan el pronóstico de la enfermedad. La cirugía programada de la raíz aórtica, especialmente con preservación valvular, permite mejorar la expectativa de vida al evitar la alta mortalidad de los eventos agudos. Alternativas prometedoras son los procederes híbridos y el intervencionismo endovascular.Marfan syndrome is a hereditary disease of the connective tissue caused by mutation of type 1 fibrillin glycoprotein-coding gene in children and adults. This disease affects organs and systems, mainly cardiovascular, skeletal, ophthalmologic systems, skin and teguments. The review of the most current aspects of diagnosis, and the multidisciplinary care to reduce morbidity and mortality of pediatric patients were presented. It was concluded that the early use of betablockers and angiotensin II AT-1 receptor blocker (losarfan are the fundamental pillars of drug therapy, since they reduce the frequency of cardiovascular complications that determine the disease prognosis. The scheduled surgery of the aortic root, particularly valve preservation, allows improving the life expectancies because it prevents high mortality from acute events. Hybrid procedures and

  4. Inhibition of HSV-1 replication by laser diode-irradiation: possible mechanism of action.

    Science.gov (United States)

    Donnarumma, G; De Gregorio, V; Fusco, A; Farina, E; Baroni, A; Esposito, V; Contaldo, M; Petruzzi, M; Pannone, G; Serpico, R

    2010-01-01

    Herpes labialis are the most frequent clinical manifestations of HSV-1 infection. Epithelial cells are able to respond to HSV-1 presence inducing the expression of IL-6, IL-1, TNF-α and IL-8. These proinflammatory cytokines have a function in the acute-phase response mediation, chemotaxis, inflammatory cell activation and antigen-presenting cells. In the human epithelial cell models, it has been demonstrated that, after an early induction of proinflammatory host response, HSV-1 down-modulates the proinflammatory cytokine production through the accumulation of two viral proteins, ICP4 and ICP27, whose transcription is induced by tegument protein VP16. These viral proteins, through the decreasing of stabilizing the mRNAs of proinflammatory genes, delay cytokine production to an extent that allows the virus to replicate. Moreover, viral transactivating proteins, ICP-0 and VP-16 induce IL-10 expression. The conventional treatment of herpes labialis involves the topical and systemic use of antiviral drugs but it is necessary to find new therapies that can act in a selective and non-cytotoxic manner in viral infection. Laser diode therapy has been considered as a non-invasive alternative treatment to the conventional treatment of herpes labialis in pain therapy, in modulation of inflammation and in wound healing. This study aims to report a possible mechanism of action of laser diode irradiation in prevention and reduction of severity of labial manifestations of herpes labialis virus. We investigated, in an in vitro model of epithelial cells HaCat, the laser-effect on HSV-1 replication and we evaluated the modulation of expression of certain proinflammatory cytokines (TNF-α, IL-1β and IL-6), antimicrobial peptide HBD2, chemokine IL-8 and the immunosuppressive cytokine, IL-10. Our results lead us to hypothesize that LD-irradiation acts in the final stage of HSV-1 replication by limiting viral spread from cell to cell and that laser therapy acts also on the host immune

  5. In vitro schistosomicidal and antiviral activities of Arctium lappa L. (Asteraceae) against Schistosoma mansoni and Herpes simplex virus-1.

    Science.gov (United States)

    Dias, Mirna Meana; Zuza, Ohana; Riani, Lorena R; de Faria Pinto, Priscila; Pinto, Pedro Luiz Silva; Silva, Marcos P; de Moraes, Josué; Ataíde, Ana Caroline Z; de Oliveira Silva, Fernanda; Cecílio, Alzira Batista; Da Silva Filho, Ademar A

    2017-10-01

    Schistosomiasis and herpes diseases represent serious issues to the healthcare systems, infecting a large number of people worldwide, mainly in developing countries. Arctium lappa L. (Asteraceae), known as "bardana" and "burdock", is a medicinal plant popularly used for several purposes, including as antiseptic. In this study, we evaluated the in vitro schistosomicidal and antiherpes activities of the crude extract of A. lappa, which have not yet been described. Fruits of A. lappa L. were extracted by maceration with ethanol: H 2 O (96:4 v/v) in order to obtain the hydroalcoholic extract of A. lappa (AL). In vitro schistosomicidal assays were assessed against adult worms of Schistosoma mansoni, while the in vitro antiviral activity of AL was evaluated on replication of Herpes simplex virus type-1 (HSV-1). Cell viability was measured by MTT assay, using Vero cells and chemical composition of AL was determined by qualitative UPLC-ESI-QTOF-MS analysis. UPLC-ESI-QTOF-MS analysis of AL revealed the presence of dibenzylbutyrolactone lignans, such as arctiin and arctigenin. Results showed that AL was not cytotoxic to Vero cells even when tested at 400μg/mL. qPCR results indicated a significant viral load decreased for all tested concentrations of AL (400, 50, and 3.125μg/mL), which showed similar antiviral effect to acyclovir (50μg/mL) when tested at 400μg/mL. Also, AL (400, 200, and 100μg/mL) caused 100% mortality and significantly reduction on motor activity of all adult worms of S. mansoni. Confocal laser scanning microscopy showed tegumental morphological alterations and changes on the numbers of tubercles of S. mansoni worms in a dose-dependent manner after treatment with AL. This report provides the first evidence for the in vitro schistosomicidal and antiherpes activities of AL, opening the route to further schistosomicidal and antiviral studies with AL and their compounds, especially lignans. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  6. A new view on the morphology and phylogeny of eugregarines suggested by the evidence from the gregarine Ancora sagittata (Leuckart, 1860 Labbé, 1899 (Apicomplexa: Eugregarinida

    Directory of Open Access Journals (Sweden)

    Timur G. Simdyanov

    2017-05-01

    . Comparative morphological analyses of gregarine teguments and attachment organelles lead us to revise their terminology. The terms “longitudinal folds” and “mucron” are restricted to archigregarines, whereas the terms “epicystic crests” and “epimerite” are proposed to describe the candidate synapomorphies of eugregarines, which, consequently, are considered as a monophyletic group. Abolishing the suborders Aseptata and Septata, incorporating neogregarines into the Eugregarinida, and treating the major molecular phylogenetic lineages of eugregarines as superfamilies appear as the best way of reconciling recent morphological and molecular evidence. Accordingly, the diagnosis of the order Eugregarinida Léger, 1900 is updated.

  7. Whole-organ isolation approach as a basis for tissue-specific analyses in Schistosoma mansoni.

    Directory of Open Access Journals (Sweden)

    Steffen Hahnel

    Full Text Available BACKGROUND: Schistosomiasis is one of the most important parasitic diseases worldwide, second only to malaria. Schistosomes exhibit an exceptional reproductive biology since the sexual maturation of the female, which includes the differentiation of the reproductive organs, is controlled by pairing. Pathogenicity originates from eggs, which cause severe inflammation in their hosts. Elucidation of processes contributing to female maturation is not only of interest to basic science but also considering novel concepts combating schistosomiasis. METHODOLOGY/PRINCIPAL FINDINGS: To get direct access to the reproductive organs, we established a novel protocol using a combined detergent/protease-treatment removing the tegument and the musculature of adult Schistosoma mansoni. All steps were monitored by scanning electron microscopy (SEM and bright-field microscopy (BF. We focused on the gonads of adult schistosomes and demonstrated that isolated and purified testes and ovaries can be used for morphological and structural studies as well as sources for RNA and protein of sufficient amounts for subsequent analyses such as RT-PCR and immunoblotting. To this end, first exemplary evidence was obtained for tissue-specific transcription within the gonads (axonemal dynein intermediate chain gene SmAxDynIC; aquaporin gene SmAQP as well as for post-transcriptional regulation (SmAQP. CONCLUSIONS/SIGNIFICANCE: The presented method provides a new way of getting access to tissue-specific material of S. mansoni. With regard to many still unanswered questions of schistosome biology, such as elucidating the molecular processes involved in schistosome reproduction, this protocol provides opportunities for, e.g., sub-transcriptomics and sub-proteomics at the organ level. This will promote the characterisation of gene-expression profiles, or more specifically to complete knowledge of signalling pathways contributing to differentiation processes, so discovering involved

  8. In Vitro Anti-Echinococcal and Metabolic Effects of Metformin Involve Activation of AMP-Activated Protein Kinase in Larval Stages of Echinococcus granulosus

    Science.gov (United States)

    Loos, Julia A.; Cumino, Andrea C.

    2015-01-01

    Metformin (Met) is a biguanide anti-hyperglycemic agent, which also exerts antiproliferative effects on cancer cells. This drug inhibits the complex I of the mitochondrial electron transport chain inducing a fall in the cell energy charge and leading 5'-AMP-activated protein kinase (AMPK) activation. AMPK is a highly conserved heterotrimeric complex that coordinates metabolic and growth pathways in order to maintain energy homeostasis and cell survival, mainly under nutritional stress conditions, in a Liver Kinase B1 (LKB1)-dependent manner. This work describes for the first time, the in vitro anti-echinococcal effect of Met on Echinococcus granulosus larval stages, as well as the molecular characterization of AMPK (Eg-AMPK) in this parasite of clinical importance. The drug exerted a dose-dependent effect on the viability of both larval stages. Based on this, we proceeded with the identification of the genes encoding for the different subunits of Eg-AMPK. We cloned one gene coding for the catalytic subunit (Eg-ampkɑ) and two genes coding for the regulatory subunits (Eg-ampkβ and Eg-ampkγ), all of them constitutively transcribed in E. granulosus protoscoleces and metacestodes. Their deduced amino acid sequences show all the conserved functional domains, including key amino acids involved in catalytic activity and protein-protein interactions. In protoscoleces, the drug induced the activation of AMPK (Eg-AMPKɑ-P176), possibly as a consequence of cellular energy charge depletion evidenced by assays with the fluorescent indicator JC-1. Met also led to carbohydrate starvation, it increased glucogenolysis and homolactic fermentation, and decreased transcription of intermediary metabolism genes. By in toto immunolocalization assays, we detected Eg-AMPKɑ-P176 expression, both in the nucleus and the cytoplasm of cells as in the larval tegument, the posterior bladder and the calcareous corpuscles of control and Met-treated protoscoleces. Interestingly, expression of Eg

  9. OVERCOMING DORMANCY IN SANSÃO-DO-CAMPO (Mimosa caesalpiniaefolia L. SEEDS SUPERAÇÃO DE DORMÊNCIA EM SEMENTES DE SANSÃO-DO-CAMPO (Mimosa caesalpiniaefolia L.

    Directory of Open Access Journals (Sweden)

    Eduardo Garcia Frasseto

    2007-09-01

    Full Text Available

    The Sansão-do-campo, also known as sabiá, is a plant whose wood has multiple uses, besides its ornamental value. It is used in forestry, for hedges and as forage. The flowers are visited by bees and tree stalk bark is used to treat lung diseases. The Sansão-do-campo seeds have shown germination problems due to tegument impermeability. To study this problem an experiment was carried out with seeds collected in August 2000 in Jussara, Goiás State. Pure seed samples were immersed in sulfuric acid (H2SO4 P. A. for 0, 5, 10, 15, and 20 minutes and then washed in running water for 10 minutes to remove acid residues. Seeds were dried and then submitted to the standard germination test. Statistical analysis showed that H2SO4 immersion for 10 to 15 minutes resulted in the highest germination rate (98%.

    KEY-WORDS: Sansão-do-campo; seeds; dormency.

    O Sansão-do-campo é uma planta cuja madeira apresenta múltiplas utilidades, além de se constituir em planta ornamental. É usado para reflorestamentos e cercas vivas, e suas folhas constituem valiosa forragem. As sementes dessa espécie, entretanto, têm apresentado dificuldades de germinação em virtude da impermeabilidade do tegumento. Para a superar o problema, instalou-se um ensaio no Laboratório de Análise de Sementes da Universidade Federal de Goiás, com sementes colhidas na cidade de Jussara, Goiás, em agosto de 2000. As amostras de sementes foram submetidas a tratamento com ácido sulfúrico (H2SO4 P. A. por períodos de 0, 5, 10, 15 e 20 minutos e, posteriormente, ao teste-padrão de germinação. A análise dos dados revelou que a imersão das sementes em H2SO4 por um período de cinco a quinze minutos produziu a maior percentagem de germinação (98%.

  10. Production and characterization of a monoclonal antibody against recombinant cathepsin L1 of Fasciola gigantica.

    Science.gov (United States)

    Anuracpreeda, Panat; Srirakam, Thippawan; Pandonlan, Sudarat; Changklungmoa, Narin; Chotwiwatthanakun, Charoonroj; Tinikul, Yotsawan; Poljaroen, Jaruwan; Meemon, Krai; Sobhon, Prasert

    2014-08-01

    Monoclonal antibodies (MoAbs) against a recombinant cathepsin L1 of Fasciola gigantica (rFgCatL1) were produced in vitro by fusion of BALB/c mice spleen cells immunized with rFgCatL1 and mouse myeloma cells. Reactivity and specificity of these MoAbs were evaluated by indirect ELISA and immunoblotting techniques. Seven MoAb clones were selected from the stable hybridoma clones, namely 1E10, 1F5, 3D11, 4B10, 4D3, 4E3 and 5E7. Clones 1E10, 1F5 and 3D11 were IgM, whereas clones 4B10, 4D3, 4E3 and 5E7 were IgG1. All MoAbs had kappa light chain isotypes. All MoAbs reacted with rCatL1 at molecular weight (MW) 30kDa and with the native CatL1 at MW 27kDa in whole body (WB) extracts of metacercariae (Met), newly excysted juveniles (NEJ), 1, 3, 5-week-old juveniles (Ju), adult WB and adult excretory-secretory (ES) fractions, but not with adult tegumental antigens (TA). All of these MoAbs showed no cross-reactions with antigens of other parasites commonly found in ruminants and human, including Paramphistomum cervi, Eurytrema pancreaticum, Gigantocotyle explanatum, Schistosoma spindale, Schistosoma mansoni, Moniezia benedeni, Avitellina centripunctata, Trichuris sp., Haemonchus placei and Setaria labiato-papillosa. Localization of CatL1 in each developmental stages of F. gigantica by immunoperoxidase technique, using these MoAbs as probes, indicated that CatL1 was present at high concentration in the caecal epithelium and caecal lumen of metacercariae, NEJ, 1, 3, 5-week-old juveniles and adult fluke. This finding indicated that CatL1 is a copiously expressed parasite protein that is released into the ES, thus CatL1 and its MoAb could be a good candidate for immunodiagnosis of fasciolosis in ruminant and human. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. Understanding the main route of drug entry in adult Fasciola hepatica: Further insights into closantel pharmacological activity.

    Science.gov (United States)

    Ceballos, L; Canton, C; Cadenazzi, G; Larsen, K; Virkel, G; Moreno, L; Fairweather, I; Lanusse, C; Alvarez, L

    2017-10-01

    Closantel (CLS) is highly effective against adult liver flukes after its oral or subcutaneous (sc) administration in ruminants. Trans-tegumental diffusion and oral ingestion are the two potential routes available for the entry of drugs into Fasciola hepatica. The work reported here contributes to improve the understanding of CLS pharmacology. The main goals of were: I) to determine the pattern of in vivo CLS accumulation into adult F. hepatica and relevant tissues in CLS-treated sheep; II) to investigate the influence of the physicochemical composition of the incubation medium on the CLS diffusion process into adult F. hepatica; III) to assess the ovicidal activity of CLS against F. hepatica eggs; and IV) to investigate the in vivo effect of CLS treatment on glutathione S-transferases activity in adult liver flukes exposed to CLS. Fourteen healthy sheep were each orally infected with 75 F. hepatica metacercariae. Sixteen (16) weeks after infection, animals were treated with CLS by oral (n = 6, 10 mg/kg) or sub-cutaneous (sc) (n = 6, 5 mg/kg) route. At 12, 24 and 36 h post-treatment, animals were sacrificed (n = 2) and samples of blood, bile and adult F. hepatica were collected. In addition, flukes recovered from non-treated sheep (n = 2) were ex vivo incubated (60 min) in the presence of CLS in either RPMI or bile as incubation medium. CLS concentration was measured by HPLC. The ovicidal activity of CLS was investigated using eggs obtained from the bile of untreated sheep. Finally, glutathione S-transferase activity in F. hepatica recovered from untreated and CLS-treated sheep was assessed. In the in vivo studies, the highest CLS concentrations were measured in plasma and adult liver flukes. A positive correlation was observed between CLS concentration in plasma and in F. hepatica. Results obtained in the current work indicate that the in vivo accumulation of CLS into adult liver flukes occurs mainly by the oral route. After ex

  12. Clonorchis sinensis acetoacetyl-CoA thiolase: identification and characterization of its potential role in surviving in the bile duct.

    Science.gov (United States)

    Lin, Jinsi; Qu, Hongling; Chen, Guishan; He, Lei; Xu, Yanquan; Xie, Zhizhi; Ren, Mengyu; Sun, Jiufeng; Li, Shan; Chen, Wenjun; Chen, Xueqing; Wang, Xiaoyun; Li, Xuerong; Liang, Chi; Huang, Yan; Yu, Xinbing

    2015-02-25

    Clonorchis sinensis (C. sinensis) inhabits in bile duct of the host. However, the mechanisms involved in why C. sinensis can survive in the bile environment containing lipids have not yet been explored. In this study, C. sinensis acetoacetyl-CoA thiolase (CsACAT), a member of the thiolase family which has a key role in the beta oxidation pathway of fatty acid production, was identified and characterized to understand its potential role in adapting to the bile environment. The encoding sequence, conserved domains and spatial structure of CsACAT were identified and analyzed by bioinformatic tools. Recombinant CsACAT (rCsACAT) was obtained using a procaryotic expression system. The expression pattern of CsACAT was confirmed by quantitative real-time PCR, western blotting, and immunofluorescence. Gradients of lecithin were then set to culture C. sinensis adults in vitro and the survival rate of C. sinensis was analyzed, as well as the expression level and enzymatic activity of CsACAT in different lipid environments. Hypercholesteremia rabbit models were established by feeding with a hyperlipidemic diet and then infected intragastrically with C. sinensis. One and a half months later, the worm burdens and the expression level of CsACAT was detected. CsACAT was confirmed to be a member of the thiolase family and present in the excretory/secretory proteins of C. sinensis. CsACAT was specifically localized at the vitellarium and sub-tegumental muscle layer in adult worms. The mRNA level of CsACAT in eggs was higher than those in adult worms and metacercariae. When adult worms were cultured with higher concentration of lecithin, the expression level and enzyme activity of CsACAT were up-regulated. The survival rate of adult worms was higher than control group. More adult worms were recovered from hypercholesteremia rabbit models. The expression level of CsACAT in these worms was higher than control group. Our results implied that C. sinensis might sense lipid levels and

  13. In silico analysis of the fucosylation-associated genome of the human blood fluke Schistosoma mansoni: cloning and characterization of the enzymes involved in GDP-L-fucose synthesis and Golgi import.

    Science.gov (United States)

    Peterson, Nathan A; Anderson, Tavis K; Wu, Xiao-Jun; Yoshino, Timothy P

    2013-07-09

    Carbohydrate structures of surface-expressed and secreted/excreted glycoconjugates of the human blood fluke Schistosoma mansoni are key determinants that mediate host-parasite interactions in both snail and mammalian hosts. Fucose is a major constituent of these immunologically important glycans, and recent studies have sought to characterize fucosylation-associated enzymes, including the Golgi-localized fucosyltransferases that catalyze the transfer of L-fucose from a GDP-L-fucose donor to an oligosaccharide acceptor. Importantly, GDP-L-fucose is the only nucleotide-sugar donor used by fucosyltransferases and its availability represents a bottleneck in fucosyl-glycotope expression. A homology-based genome-wide bioinformatics approach was used to identify and molecularly characterize the enzymes that contribute to GDP-L-fucose synthesis and Golgi import in S. mansoni. Putative functions were further investigated through molecular phylogenetic and immunocytochemical analyses. We identified homologs of GDP-D-mannose-4,6-dehydratase (GMD) and GDP-4-keto-6-deoxy-D-mannose-3,5-epimerase-4-reductase (GMER), which constitute a de novo pathway for GDP-L-fucose synthesis, in addition to a GDP-L-fucose transporter (GFT) that putatively imports cytosolic GDP-L-fucose into the Golgi. In silico primary sequence analyses identified characteristic Rossman loop and short-chain dehydrogenase/reductase motifs in GMD and GMER as well as 10 transmembrane domains in GFT. All genes are alternatively spliced, generating variants of unknown function. Observed quantitative differences in steady-state transcript levels between miracidia and primary sporocysts may contribute to differential glycotope expression in early larval development. Additionally, analyses of protein expression suggest the occurrence of cytosolic GMD and GMER in the ciliated epidermal plates and tegument of miracidia and primary sporocysts, respectively, which is consistent with previous localization of highly

  14. In Vitro Replication of Chelonid Herpesvirus 5 in Organotypic Skin Cultures from Hawaiian Green Turtles (Chelonia mydas).

    Science.gov (United States)

    Work, Thierry M; Dagenais, Julie; Weatherby, Tina M; Balazs, George H; Ackermann, Mathias

    2017-09-01

    Fibropapillomatosis (FP) is a tumor disease of marine turtles associated with chelonid herpesvirus 5 (ChHV5), which has historically been refractory to growth in tissue culture. Here we show, for the first time, de novo formation of ChHV5-positive intranuclear inclusions in cultured green turtle cells, which is indicative of active lytic replication of the virus. The minimal requirements to achieve lytic replication in cultured cells included (i) either in vitro cultures of ChHV5-positive tumor biopsy specimens (plugs) or organotypic cultures (rafts) consisting of ChHV5-positive turtle fibroblasts in collagen rafts seeded with turtle keratinocytes and (ii) keratinocyte maturation induced by raising raft or biopsy cultures to the air-liquid interface. Virus growth was confirmed by detailed electron microscopic studies that revealed intranuclear sun-shaped capsid factories, tubules, various stages of capsid formation, nuclear export by budding into the perinuclear space, tegument formation, and envelopment to complete de novo virus production. Membrane synthesis was also observed as a sign of active viral replication. Interestingly, cytoplasmic particles became associated with keratin filaments, a feature not seen in conventional monolayer cell cultures, in which most studies of herpesvirus replication have been performed. Our findings draw a rich and realistic picture of ChHV5 replication in cells derived from its natural host and may be crucial not only to better understand ChHV5 circulation but also to eventually complete Koch's postulates for FP. Moreover, the principles described here may serve as a model for culture of other viruses that are resistant to replication in conventional cell culture. IMPORTANCE A major challenge in virology is the study of viruses that cannot be grown in the laboratory. One example is chelonid herpesvirus 5 (ChHV5), which is associated with fibropapillomatosis, a globally distributed, debilitating, and fatal tumor disease of

  15. A Simple Proteomics-Based Approach to Identification of Immunodominant Antigens from a Complex Pathogen: Application to the CD4 T Cell Response against Human Herpesvirus 6B.

    Directory of Open Access Journals (Sweden)

    Aniuska Becerra-Artiles

    Full Text Available Most of humanity is chronically infected with human herpesvirus 6 (HHV-6, with viral replication controlled at least in part by a poorly characterized CD4 T cell response. Identification of viral epitopes recognized by CD4 T cells is complicated by the large size of the herpesvirus genome and a low frequency of circulating T cells responding to the virus. Here, we present an alternative to classical epitope mapping approaches used to identify major targets of the T cell response to a complex pathogen like HHV-6B. In the approach presented here, extracellular virus preparations or virus-infected cells are fractionated by SDS-PAGE, and eluted fractions are used as source of antigens to study cytokine responses in direct ex vivo T cell activation studies. Fractions inducing significant cytokine responses are analyzed by mass spectrometry to identify viral proteins, and a subset of peptides from these proteins corresponding to predicted HLA-DR binders is tested for IFN-γ production in seropositive donors with diverse HLA haplotypes. Ten HHV-6B viral proteins were identified as immunodominant antigens. The epitope-specific response to HHV-6B virus was complex and variable between individuals. We identified 107 peptides, each recognized by at least one donor, with each donor having a distinctive footprint. Fourteen peptides showed responses in the majority of donors. Responses to these epitopes were validated using in vitro expanded cells and naturally expressed viral proteins. Predicted peptide binding affinities for the eight HLA-DRB1 alleles investigated here correlated only modestly with the observed CD4 T cell responses. Overall, the response to the virus was dominated by peptides from the major capsid protein U57 and major antigenic protein U11, but responses to other proteins including glycoprotein H (U48 and tegument proteins U54 and U14 also were observed. These results provide a means to follow and potentially modulate the CD4 T-cell immune

  16. Human Asymptomatic Epitope Peptide/CXCL10-Based Prime/Pull Vaccine Induces Herpes Simplex Virus-Specific Gamma Interferon-Positive CD107+ CD8+ T Cells That Infiltrate the Cornea and Trigeminal Ganglia of Humanized HLA Transgenic Rabbits and Protect against Ocular Herpes Challenge.

    Science.gov (United States)

    Khan, Arif A; Srivastava, Ruchi; Vahed, Hawa; Roy, Soumyabrata; Walia, Sager S; Kim, Grace J; Fouladi, Mona A; Yamada, Taikun; Ly, Vincent T; Lam, Cynthia; Lou, Anthony; Nguyen, Vivianna; Boldbaatar, Undariya; Geertsema, Roger; Fraser, Nigel W; BenMohamed, Lbachir

    2018-06-13

    Herpes simplex virus 1 (HSV-1) is a prevalent human pathogen that infects the cornea causing potentially blinding herpetic disease. A clinical herpes vaccine is still lacking. In the present study, a novel prime/pull vaccine was tested in Human Leukocyte Antigen- (HLA-) transgenic rabbit model of ocular herpes (HLA Tg rabbit). Three asymptomatic (ASYMP) peptide epitopes were selected from the HSV-1 membrane glycoprotein C (UL44 400-408 ), the DNA replication binding helicase (UL9 196-204 ), and the tegument protein (UL25 572-580 ), all preferentially recognized by CD8 + T cells from "naturally protected" HSV-1-seropositive healthy ASYMP individuals (who never had recurrent corneal herpetic disease). HLA Tg rabbits were immunized with a mixture of these three ASYMP CD8 + T cell peptide epitopes (UL44 400-408 , UL9 196-204 and UL25 572-580 ), delivered subcutaneously with CpG 2007 adjuvant (prime). Fifteen days later, half of the rabbits received a topical ocular treatment with a recombinant neurotropic AAV8 vector, expressing the T cell-attracting CXCL10 chemokine (pull). The frequency, function of HSV-specific CD8 + T cells induced by the prime/pull vaccine were assessed in peripheral blood, cornea, and trigeminal ganglia (TG). Compared to peptides alone, the peptides/CXCL10 prime/pull vaccine generated frequent polyfunctional gamma interferon-positive (IFN-γ + ) CD107 + CD8 + T cells that infiltrated both the cornea and TG. CD8 + T cells mobilization into cornea and TG of prime/pull- vaccinated rabbits was associated with a significant reduction in corneal herpes infection and disease following an ocular HSV-1 challenge (McKrae). These findings draw attention to the novel prime/pull vaccine strategy to mobilize anti-viral CD8 + T cells into tissues protecting them against herpes infection and disease. IMPORTANCE There is an urgent need for a vaccine against widespread herpes simplex virus infections. The present study demonstrates that immunization of HLA

  17. Schistosome syntenin partially protects vaccinated mice against Schistosoma mansoni infection.

    Directory of Open Access Journals (Sweden)

    Barbara C Figueiredo

    2014-08-01

    Full Text Available Schistosomiasis is a neglected tropical disease caused by several species of trematode of the genus Schistosoma. The disease affects more than 200 million people in the world and causes up to 280,000 deaths per year, besides having high morbidity due to chronic illness that damages internal organs. Current schistosomiasis control strategies are mainly based on chemotherapy, but many researchers believe that the best long-term strategy to control disease is a combination of drug treatment and immunization with an anti-schistosome vaccine. Among the most promising molecules as vaccine candidates are the proteins present in the tegument and digestive tract of the parasite.In this study, we describe for the first time Schistosoma mansoni syntenin (SmSynt and we evaluate its potential as a recombinant vaccine. We demonstrate by real-time PCR that syntenin is mainly expressed in intravascular life stages (schistosomula and adult worms of the parasite life cycle and, by confocal microscopy, we localize it in digestive epithelia in adult worms and schistosomula. Administration of siRNAs targeting SmSynt leads to the knock-down of syntenin gene and protein levels, but this has no demonstrable impact on parasite morphology or viability, suggesting that high SmSynt gene expression is not essential for the parasites in vitro. Mice immunization with rSmSynt, formulated with Freund's adjuvant, induces a Th1-type response, as suggested by the production of IFN-γ and TNF-α by rSmSynt-stimulated cultured splenocytes. The protective effect conferred by vaccination with rSmSynt was demonstrated by 30-37% reduction of worm burden, 38-43% reduction in the number, and 35-37% reduction in the area, of liver granulomas.Our report is the first characterization of syntenin in Schistosoma mansoni and our data suggest that this protein is a potential candidate for the development of a multi-antigen vaccine to control schistosomiasis.

  18. In vivo activity of albendazole in combination with thymol against Echinococcus multilocularis.

    Science.gov (United States)

    Albani, Clara María; Pensel, Patricia Eugenia; Elissondo, Natalia; Gambino, Guillermo; Elissondo, María Celina

    2015-09-15

    Human alveolar echinococcosis (AE) is caused by the fox tapeworm Echinococcus multilocularis and is usually lethal if left untreated. The current strategy for treating human AE is surgical resection of the parasite mass complemented by chemotherapy with benzimidazole compounds. However, reliable chemotherapeutic alternatives have not yet been developed stimulating the research of new treatment strategies such as the use of medicinal plants. The aim of the current study was to investigate the efficacy of the combination albendazole (ABZ)+thymol on mice infected with E. multilocularis metacestodes. For this purpose, mice infected with parasite material were treated daily for 20 days with ABZ (5 mg/kg), thymol (40 mg/kg) or ABZ (5 mg/kg)+thymol (40 mg/kg) or left untreated as controls. After mice were euthanized, cysts were removed from the peritoneal cavity and the treatment efficacy was evaluated by the mean cysts weight, viability of protoscoleces and ultrastructural changes of cysts and protoscoleces. The application of thymol or the combination of ABZ+thymol resulted in a significant reduction of the cysts weight compared to untreated mice. We also found that although ABZ and thymol had a scolicidal effect, the combination of the two compounds had a considerably stronger effect showing a reduction in the protoscoleces viability of 62%. These results were also corroborated by optical microscopy, SEM and TEM. Protoscoleces recovered from ABZ or thymol treated mice showed alterations as contraction of the soma region, rostellar disorganization and presence of blebs in the tegument. However both drugs when combined lead to a total loss of the typical morphology of protoscoleces. All cysts removed from control mice appeared intact and no change in ultrastructure was detected. In contrast, cysts developed in mice treated with ABZ revealed changes in the germinal layer as reduction in cell number, while the treatment with thymol or the ABZ+thymol combination

  19. GPR Prospecting and Endoscopic Investigation in a Renaissance Church

    Science.gov (United States)

    Persico, Raffaele; Matera, Loredana; Bianco, Nadia; Masini, Nicola; Leopizzi, Giuseppe

    2015-04-01

    Nondestructive Evaluation, vol. 17, pp. 71-83, 200

  20. Germination of seeds of the invasive plant Sesbania virgata (cav. pers. under effects of light, temperature, and dormancy overcoming Germinação de sementes da invasora Sesbania virgata (cav. pers. sob efeito de luz, temperatura e superação de dormência

    Directory of Open Access Journals (Sweden)

    Irenice Gomes de Oliveira

    2010-12-01

    Full Text Available The work had as objectives to evaluate the influence of the light, temperature, and chemical scarification on the germination of seeds of S. virgata, species still little studied and that is invasive in several niches of the caatinga ecosystem. The seeds of S. virgata were collected in the municipal district of Natuba-PB in March/2008. Eight treatments were used as follows: intact seeds in the presence and absence of light, at 25ºC; intact seeds in the presence and absence of light, at 30ºC; submerged seeds in sulfuric acid for five minutes in the presence and absence of light, at 25ºC; submerged seeds in sulfuric acid for five minutes in the presence and absence of light, at 30ºC. The germination tests were accomplished in germination cameras under light and continuous darkness at constant temperatures of 25ºC and 30ºC, in a 12 hours photoperiod. The seeds were put to germinate in plastic boxes (gerbox with filter paper. The seeds of S. virgata have tegument dormancy and show low germination tax when intact, which is a strategy for seed banks formation. The acid scarification increased germination percentage and speed germination index. Este trabalho teve como objetivos avaliar a influência da luz, da temperatura e da escarificação química sobre a germinação de sementes de S. virgata, espécie ainda pouco estudada e que se constata invasora em diversos nichos do bioma Caatinga. Sementes de S. virgata foram coletadas no município de Natuba-PB em março de 2008 e submetidas a oito tratamentos, descritos a seguir: sementes intactas, na presença e ausência de luz, a 25ºC; sementes intactas na presença e ausência de luz, a 30ºC; sementes imersas em ácido sulfúrico por cinco minutos na presença e ausência de luz, a 25ºC; sementes imersas em ácido sulfúrico por cinco minutos na presença e ausência de luz, a 30ºC. Para estimar os efeitos dos tratamentos foram realizados testes de germinação em câmaras de germinação sob

  1. Germinação in vitro de eixos embrionários zigóticos de imbuia (ocotea porosa (NEES EX MARTIUS Liberato Barroso In vitro germination of zygotic embryonic axes of imbuia (Ocotea porosa (NEES EX MARTIUS liberato barroso

    Directory of Open Access Journals (Sweden)

    Luciana Luiza Pelegrini

    2013-04-01

    Full Text Available A imbuia apresenta sementes recalcitrantes, com forte dormência tegumentar, irregularidade e baixa germinação, dificultando sua propagação natural. O objetivo deste trabalho foi avaliar a germinação in vitro de eixos embrionários de imbuia, testando o efeito de diferentes concentrações de sacarose, de carvão ativado e de formulações salinas para acelerar a produção de mudas. Eixos embrionários zigóticos foram inoculados em meio de cultura MS/2 contendo diferentes concentrações de sacarose (15, 30, 60 ou 90 g L-1, de carvão ativado (0, 1, 2 ou 3 g L-1 e diferentes formulações salinas (MS, WPM, MS/2 ou WPM/2. Os resultados indicaram que a adição de 15 g L-1 de sacarose foi insuficiente para promover a germinação dos eixos embrionários e as demais concentrações promoveram entre 45,5 e 59,4% de germinação. Com relação às formulações salinas, os melhores resultados foram obtidos com os meios de cultura MS e MS/2 obtendo-se 67,7 e 74,2% de germinação, respectivamente. A adição de carvão ativado favoreceu a germinação in vitro dos eixos embrionários, enquanto que na ausência de carvão ativado não ocorreu germinação e houve elevada porcentagem de oxidação. A germinação in vitro de imbuia é viável e pode ser obtida com eixos embrionários zigóticos maduros com a formulação salina MS/2, suplementada com 30 g L-1 de sacarose e 1g L-1 de carvão ativado.Imbuia has recalcitrant seeds with strong tegumental dormancy, irregular and low germination, which impair natural propagation. The objective of this study was to evaluate the in vitro germination of imbuia embryonic axes by testing the effect of different concentrations of sucrose, activated charcoal and salt formulations in order to accelerate the production of plants. Zygotic embryonic axes were inoculated in MS/2 culture medium with different concentrations of sucrose (15, 30, 60 or 90 g L-1, of activated charcoal (0, 1, 2 or 3 gL-1 and in

  2. Dormancy breaking in Mauritia flexuosa seeds by using mechanical scarification and soaking = Superação de dormência em sementes de buriti por meio da escarificação mecânica e embebição

    Directory of Open Access Journals (Sweden)

    Alexsander Seleguini

    2012-12-01

    Full Text Available The majority of Arecaceae species presents difficulties to germinate, even under appropriate conditions, complicating the seedlings production process. This study aimed at evaluating the influence of seed scarification and soaking in the emergence and development of Mauritia flexuosa seedlings. The treatments consisted of a combination of seed soaking and scarification methods, in a completely randomized design, in a 2 x 3 + 1 factorial scheme, with five replications. Seeds with and without scarification, combined with soaking methods (not soaked seeds and seeds soaked for 30 days, with and without daily water renewal, plus an additional treatment, where the seeds were scarified after soaking for 30 days, were tested. The seedlingemergence up to 146 days after sowing (d.a.s. was evaluated, as well as the emergence rate index (46 and 146 d.a.s. and growth components (146 d.a.s.. The results were submitted to the variance analysis (F test and means compared by using the Tukey test, at 5%. It was possible to conclude that the Mauritia flexuosa seeds presented tegument dormancy. The seed scarification increased the germination rate, however, it contributed to increase the seeds mortality rate. The soaking of nonscarified seeds for 30 days, with daily water renewal, improved the potential for seed germination. The mechanical scarification, without or after soaking the seeds in water, increased the Mauritia flexuosa seedlings mortality rate, and it is not, therefore, a suitable method for dormancy breaking. = A maioria das espécies de Arecaceae apresenta dificuldades para germinar, mesmo sob condições adequadas, dificultando a produção de mudas. Objetivou-se, com este trabalho, avaliar a influência da escarificação e embebição de sementes na emergência e desenvolvimento de mudas de buritizeiro. Os tratamentos foram compostos pela combinação de métodos de embebição e escarificação de sementes, adotando-se o delineamento inteiramente

  3. Caracterização morfológica e química de frutos de rambutan Morphologic and chemical characterization of rambutan fruits

    Directory of Open Access Journals (Sweden)

    Renata Aparecida de Andrade

    2008-12-01

    genotype distinction and the genetic distance among the materials. It was evaluated: fruit size, filament length, peel thickness, seed size, fruit, peel and seed weight, pulp percentage, soluble solid contents (ºBrix, ascorbic acid and titrable acidity. Morever, the following observations were conducted: peel coloration, filaments and pulp, adherence of the pulp to the peel, adherence of the pulp to the seed and presence of tegument of the seed in the pulp. Based on the obtained results in this research, it can be concluded that the materials A51 and B03 present a better group of desirable characteristics, revealing to be promising for the producers in the installation of new culture plantings.

  4. Prophylactic Herpes Simplex Virus 2 (HSV-2) Vaccines Adjuvanted with Stable Emulsion and Toll-Like Receptor 9 Agonist Induce a Robust HSV-2-Specific Cell-Mediated Immune Response, Protect against Symptomatic Disease, and Reduce the Latent Viral Reservoir.

    Science.gov (United States)

    Hensel, Michael T; Marshall, Jason D; Dorwart, Michael R; Heeke, Darren S; Rao, Eileen; Tummala, Padmaja; Yu, Li; Cohen, Gary H; Eisenberg, Roselyn J; Sloan, Derek D

    2017-05-01

    Several prophylactic vaccines targeting herpes simplex virus 2 (HSV-2) have failed in the clinic to demonstrate sustained depression of viral shedding or protection from recurrences. Although these vaccines have generated high titers of neutralizing antibodies (NAbs), their induction of robust CD8 T cells has largely been unreported, even though evidence for the importance of HSV-2 antigen-specific CD8 T cells is mounting in animal models and in translational studies involving subjects with active HSV-2-specific immune responses. We developed a subunit vaccine composed of the NAb targets gD and gB and the novel T cell antigen and tegument protein UL40, and we compared this vaccine to a whole-inactivated-virus vaccine (formaldehyde-inactivated HSV-2 [FI-HSV-2]). We evaluated different formulations in combination with several Th1-inducing Toll-like receptor (TLR) agonists in vivo In mice, the TLR9 agonist cytosine-phosphate-guanine (CpG) oligodeoxynucleotide formulated in a squalene-based oil-in-water emulsion promoted most robust, functional HSV-2 antigen-specific CD8 T cell responses and high titers of neutralizing antibodies, demonstrating its superiority to vaccines adjuvanted by monophosphoryl lipid A (MPL)-alum. We further established that FI-HSV-2 alone or in combination with adjuvants as well as adjuvanted subunit vaccines were successful in the induction of NAbs and T cell responses in guinea pigs. These immunological responses were coincident with a suppression of vaginal HSV-2 shedding, low lesion scores, and a reduction in latent HSV-2 DNA in dorsal root ganglia to undetectable levels. These data support the further preclinical and clinical development of prophylactic HSV-2 vaccines that contain appropriate antigen and adjuvant components responsible for programming elevated CD8 T cell responses. IMPORTANCE Millions of people worldwide are infected with herpes simplex virus 2 (HSV-2), and to date, an efficacious prophylactic vaccine has not met the rigors

  5. Cultivo de embriões imaturos de citros em diferentes concentrações de carvão ativado e ácido giberélico Activated charcoal and giberellic acid concentrations on immature embryos culture

    Directory of Open Access Journals (Sweden)

    Edvan Alves Chagas

    2005-12-01

    Full Text Available Adição de carvão ativado e giberelina no meio de cultura podem proporcionar melhores condições no desenvolvimento de embriões imaturos de citros. Objetivou-se avaliar o efeito de carvão ativado e GA3 (ácido giberélico no cultivo de embriões imaturos provenientes do cruzamento entre laranjeira 'Pêra Rio' x tangerineira 'Poncã'. Após 118 dias da polinização, frutos imaturos, com 3 a 4 cm de diâmetro, foram coletados, suas sementes removidas e tratadas com álcool (70% por cinco minutos, hipoclorito de sódio (2% por 20 minutos e, posteriormente, lavadas três vezes em água destilada e autoclavada. Em condições assépticas, os tegumentos das sementes foram separados, os embriões globulares excisados e inoculados em tubos de ensaio contendo 15 mL do meio MT, acrescido de carvão ativado (0; 0,5; 1; 1,5 e 2 g L-1 e GA3 (0; 0,01; 0,1; 1 e 10 mg L-1. Após a inoculação, os embriões permaneceram por 90 dias em sala de crescimento a 27+1ºC, fotoperíodo de 16 horas e irradiância de 32 mmol m-2 s-1. Maior comprimento da parte aérea foi obtido em meio MT, acrescido de 0,1 e 1 mg L-1 de GA3, combinado com 2 g L-1 de carvão ativado. Maior comprimento do sistema radicular, massa da matéria fresca e número de folhas de plântulas foram obtidos em meio MT, acrescido de 0,01 mg L-1 de GA3, na ausência de carvão ativado. A adição de carvão ativado influenciou na concentração de ácido giberélico acrescido no meio de cultura.Activated charcoal and gibberelin provides better conditions on development of citrus immature embryos. Activated charcoal and GA3 (gibberelic acid on 'Pêra Rio' sweet orange x 'Poncã' mandarin immature embryos culture was evaluated. After 118 days-pollination, imature fruits with 3 to 4 cm of diameter were collected, seeds removed and treated with alcohol (70% for five min., sodium hypoclorite (2% for 20 min. and three times washed with distilled and autoclaved water. In aseptic conditions, the teguments

  6. Métodos para quebra de dormência de unidades de dispersão de Zizyphus joazeiro Mart. (Rhamnaceae Methods for dormancy breaking of Zizyphus joazeiro Mart. dispersal units

    Directory of Open Access Journals (Sweden)

    Edna Ursulino Alves

    2008-06-01

    . The evaluated variables included emergence percentage, first count and emergence speed index, length and weight of plant dry matter. The treatments showing higher Z. joazeiro seedling emergence were immersion of dispersal units in cold water for 48 hours, immersion in water at 70 C for three minutes and manual scarification with sandpaper, overcoming tegument hardness in dispersal units of this species.

  7. Clonorchis sinensis granulin: identification, immunolocalization, and function in promoting the metastasis of cholangiocarcinoma and hepatocellular carcinoma.

    Science.gov (United States)

    Wang, Caiqin; Lei, Huali; Tian, Yanli; Shang, Mei; Wu, Yinjuan; Li, Ye; Zhao, Lu; Shi, Mengchen; Tang, Xin; Chen, Tingjin; Lv, Zhiyue; Huang, Yan; Tang, Xiaoping; Yu, Xinbing; Li, Xuerong

    2017-05-25

    Long-term infections by Clonorchis sinensis are associated with cholangitis, cholecystitis, liver fibrosis, cirrhosis, and even liver cancer. Molecules from the worm play vital roles in disease progress. In the present study, we identified and explored molecular characterization of C. sinensis granulin (CsGRN), a growth factor-like protein from C. sinensis excretory/secretory products (CsESPs). The encoding sequence and conserved domains of CsGRN were identified and analysed by bioinformatics tools. Recombinant CsGRN (rCsGRN) protein was expressed in Escherichia coli BL21 (DE3). The localisation of CsGRN in adult worms and Balb/c mice infected with C. sinensis was investigated by immunofluorescence and immunohistochemistry, respectively. Stable CsGRN-overexpressed cell lines of hepatoma cells (PLC-GRN cells) and cholangiocarcinoma cells (RBE-GRN cells) were constructed by transfection of eukaryotic expression plasmid of pEGFP-C1-CsGRN. The effects on cell migration and invasion of CsGRN were assessed through the wound-healing assay and transwell assay. The levels of matrix metalloproteinase 2 and 9 (MMP2 and MMP9) in PLC-GRN or RBE-GRN cells were detected by real-time PCR (qRT-PCR). The levels of E-cadherin, vimentin, N-cadherin, zona occludens proteins (ZO-1), β-catenin, phosphorylated ERK (p-ERK) and phosphorylated AKT (p-AKT) were analysed by Western blotting. CsGRN, including the conserved GRN domains, was confirmed to be a member of the granulin family. CsGRN was identified as an ingredient of CsESPs. CsGRN was localised in the tegument and testes of the adult worm. Furthermore, it appeared in the cytoplasm of hepatocytes and biliary epithelium cells from infected Balb/c mouse. The enhancement of cell migration and invasion of PLC-GRN and RBE-GRN cells were observed. In addition, CsGRN upregulated the levels of vimentin, N-cadherin, β-catenin, MMP2 and MMP9, while it downregulated the level of ZO-1 in PLC-GRN/RBE-GRN cells. In total proteins of liver tissue

  8. Número necessário de experimentos para a comparação de cultivares de feijão Number of necessary experiments for the comparison in common bean cultivars

    Directory of Open Access Journals (Sweden)

    Alberto Cargnelutti Filho

    2006-12-01

    Full Text Available O objetivo deste trabalho foi determinar o número de medições (experimentos necessários à predição do desempenho de cultivares de feijão (Phaseolus vulgaris L.. Quatorze cultivares de feijão foram avaliadas em nove experimentos conduzidos em Santa Maria, Estado do Rio Grande do Sul (latitude 29°42'S, longitude 53°49'W e 95m de altitude, nos anos agrícolas de 2000/2001 a 2004/2005. As avaliações foram constituídas por produtividade de grãos, número de vagens por planta, número de sementes por vagem, massa de cem grãos, população final de plantas, número de dias da emergência ao florescimento, número de dias da emergência à colheita, altura de inserção da primeira vagem, altura de inserção da última vagem, grau de acamamento e coloração do tegumento dos grãos. As estimativas dos coeficientes de repetibilidade foram obtidas por três métodos - análise de variância, componentes principais e análise estrutural. Sete experimentos possibilitam a identificação de cultivares superiores de feijão em relação às características de produção, de fenologia e de morfologia, com 85% de exatidão no prognóstico de seu valor real.The objective of this research was to determine how many evaluations (experiments were necessary to predict the performance of common bean (Phaseolus vulgaris cultivars. Fourteen common beans cultivars were evaluated in nine experiments conducted in Santa Maria, in Rio Grande do Sul State, Brazil (latitude 29°42'S, longitude 53°49'W, altitude 95m in agricultural years from 2000/2001 to 2004/2005. The evaluations were constituted by grain yield, number of pods per plant, number of seeds per pod, weight of 100 grains, final population of plants, number of days of the emergency to flowering, number of days of the emergency to harvest, height of first pod insertion, height of the final pod insertion, degree of the down and colour of grain tegument. Repeatability estimations were obtained

  9. Superação da dormência e avaliação da qualidade fisiológica de sementes de Sesbania virgata Dormancy break and evaluation of physiological quality of Sesbania virgata seeds

    Directory of Open Access Journals (Sweden)

    Valquíria Nogueira Camargos

    2008-12-01

    tegument cuts, followed by germination tests. To characterize lots and adapt a tetrazolium-test, in a second stage, seeds were sanded in the area opposite to the embryonic axis and submitted to the following tests: tetrazolium, germination, vigor index, fresh and dry weight. Two preconditioning conditions (soaked paper for 18 h at 30 ºC and immersion in water for 12 h at 30 ºC and three immersion timespan (one, two and three hours in tetrazolium solution 0.5 % were used. In the S. virgata lot profile analysis, variations were detected in its physiological quality according to vigor index and germination results. Scarification method with sandpaper better broke S. virgata seed dormancy, and preconditioning seeds in soaked paper followed by immersion in tetrazolium solution for two hours was the appropriate procedure to assess seed viability.

  10. Alterações morfológicas induzidas por butirato, propionato e lactato sobre a mucosa ruminal e epiderme de bezerros: II. Aspectos ultra-estruturais Lactate, propionate, and butyrate induced morphological alterations on calf ruminal mucosa and epidermis: II. Ultra-structurals aspects

    Directory of Open Access Journals (Sweden)

    S.F. Costa

    2008-02-01

    Full Text Available Avaliou-se o efeito de ácidos graxos voláteis (AGV sobre a integridade do epitélio no rúmen, no plano nasolabial, na epicera e no perioplum traseiro e dianteiro de bezerros e validou-se a feitura de biópsias tegumentares como indicadores de alterações morfológicas da mucosa ruminal. Dezessete bezerros, com sonda no rúmen, receberam infusões intra-ruminais de AGV ou salina, durante 37 dias. Aos 89 dias de vida, após o abate, foram colhidas amostras dos tecidos. Os AGV aumentaram a área de epitélio total e a área de células metabolicamente ativas no epitélio ruminal, embora o butirato não tenha induzido ao desenvolvimento papilar. A área de epitélio não queratinizado no plano nasolabial foi reduzida pela infusão de AGV. Butirato e lactato foram mais indutores de alterações patológicas no epitélio ruminal. Não foram observadas lesões histológicas nos epitélios do plano nasolabial, da epicera e do perioplum, mostrando que essas são conseqüências do efeito direto dos AGV sobre o epitélio ruminal. Os efeitos indireto e direto dos AGV sobre a morfologia dos tecidos epiteliais queratinizados não foram iguais. Biópsias tegumentares podem ter utilidade como indicadores de alterações morfológicas da mucosa ruminal.The effect of volatile fatty acids (VFA on rumen wall, epidermis of nasolabial surface, perioplum, and epicera of calves was evaluated. The experiment also aimed to validate the procedure of tegument biopsies as indicators of ruminal mucosa alterations. Seventeen neonatal calves with foley catheters received intraruminal infusions of VFA or saline, during 37 days. At 89-day-old, the animals were slaughtered and tissue samples were collected from rumen, nasolabial surface, epicera, and perioplum from face and hindquarters. VFA infusion increased total epithelium area and metabolically active ruminal cell area; although butirate did not induce the papilar development. The effect of nasolabial surface VFA

  11. Eventos externos e internos da infecção de larvas e ninfas de Rhipicephalus sanguineus por Metarhizium anisopliae External and internal events of Rhipicephalus sanguineus larvae and nymphs infection by Metarhizium anisopliae

    Directory of Open Access Journals (Sweden)

    M.V. Garcia

    2008-08-01

    than 18 hours post-inoculation and the fungus penetration through the tegument into the larvae and nymphs in, respectively, two and three days post-infection. Following penetration, the fungus invaded the body of the ticks and colonized it diffusely without a noticeable predilection for tissue, but no apparent lesions were observed. Death of larvae and nymphs occurred on the 3rd and 4th post-infection days and pathogen sporulation over the dead tick began on the 6th post-infection day.

  12. Superação de dormência de sementes como estratégia para restauração florestal de pastagem tropical Seed dormancy overcoming as a strategy for forest restoration in tropical pasture

    Directory of Open Access Journals (Sweden)

    Silvia Rahe Pereira

    2013-02-01

    H. stigonocarpa, mechanical scarification; C. langsdorffii and M. caesapiniifolia, chemical scarification with H2SO4; P. nitens, tegument puncture; and P. dubium, immersion in water at 80ºC. Recruitment of plants in the field were followed up for 21 months. Dormancy overcoming promoted greater emergency in four out of the six tested species, in the greenhouse, and in five species in the field. Furthermore, it reduced the emergency timing of the species and increased the recruitment of M. caesalpiniifolia, P. nitens, D. mollis e H. stigonocarpa, at 127 days, and of H. stigonocarpa at 659 days. Seed dormancy overcoming treatments can increase the efficiency of direct seeding in restoration projects of degraded areas in the Cerrado, promoting greater seed emergency and the recruitment of plants.

  13. Genitores potenciais para hibridações identificados por divergência genética em feijão carioca Bean parents for hybridization identified by genetic divergence in "carioca" bean

    Directory of Open Access Journals (Sweden)

    Nerinéia Dalfollo Ribeiro

    2003-06-01

    Full Text Available Noventa genótipos de feijão carioca (Phaseolus vulgaris L. foram avaliados, em dois anos agrícolas, em Santa Maria, RS, a fim de definir quais características agromorfológicas constituem-se como melhores descritores, realizar agrupamento em função de dissimilaridade genética e de definir quais combinações híbridas mais promissoras serão obtidas para o desenvolvimento de populações segregantes. Dos 20 caracteres agromorfológicos avaliados, apenas nove (ferrugem nos legumes, acamamento, nota geral, cor do tegumento, rendimento de grãos, massa de 100 sementes, altura de inserção do primeiro legume, altura de inserção do último legume e número de sementes por legume apresentaram maior contribuição para a divergência genética. Os genótipos de feijão carioca foram agrupados pelo método hierárquico de ligação completa. Populações segregantes, com variabilidade genética superior, podem ser obtidas com hibridações entre o genótipo ESAL 550 com genótipos do grupo 2 (LH-6, 17-4-32, R-78, H-4-5 e R-102 e/ou com genótipos do grupo 3 (FT 97-188, Cati-Taquari, CII-328, Carioca Precoce, FT 97-41, LH-11, FT 91-4067, Iapar 31, CI 102, Carioca MG, CII-54 e R-102.Carioca bean genotypes (Phaseolus vulgaris L. were evaluated in two growing seasons in Santa Maria, RS, Brazil. The objectives of this work were to evaluate which morpho-agronomic characteristics were the best descriptors, to group the genotypes in relation to genetic diversity and to determine which hybrid combinations are promissing to obtain higher segregation populations in carioca bean. From the 20 morpho-agronomic characteristics evaluated, only seven (pod rust, lodging, general note, colour of seed tegument, grain yield, 100 seed weight, height of first and final pod insertion and number of seeds per pod showed higher contribution to genetic diversity. The evaluated carioca bean genotypes were clustered by the complete linkage method. The following hybrid

  14. Una nueva especie de Meloe Linnaeus, 1758 del suroeste de Marruecos incluida en el grupo de M. (Eurymeloe rugosus Marsham, 1802 (Coleoptera: Meloidae

    Directory of Open Access Journals (Sweden)

    Ruiz, José L.

    2015-06-01

    Full Text Available The scientific collections of the Museo Nacional de Ciencias Naturales (MNCN-CSIC, Madrid hold an extense set of entomological materials collected in Morocco along the first decades of the XXth century by the preeminent naturalist M. Martínez de la Escalera. Morphological studies of the specimens of the genus Meloe Linnaeus, 1758 (Coleoptera: Meloidae reveals the existence of populations morphologically differentiated along the coastal regions of Essaouira and Ifni. These populations are included within the Meloe rugosus Marsham, 1802 species group in the subgenus Eurymeloe Reitter, 1911. Their differential traits with respect to all other North African and European species of the Meloe rugosus species group are constant, and permit considering these populations as a taxonomic independent unit described herein, Meloe baamarani n. sp. This new species is characterized by having a black, opaque, general coloration all over the body and appendages; black short vestiture; broad head with broadly rounded temples, without median longitudinal groove; long antennae, with segments III to VIII subcylindrical and longer than wide; pronotum transverse, with convergent sides toward the base, without median groove; head and pronotum punctuation dense; aedeagus narrow, median lobe wide and strong, dorsally sinuous, with ventral hooks close to the apex. Meloe baamarani can be only confused on the western regions of northern Africa with Meloe mediterraneus Müller, 1925. This species shares a general appearance with M. baamarani, but differs in many morphological traits. Among those, tegument micro-reticulation, absence of median groove along the head, pronotum morphology and macrosculpture, and configuration of the male genitalia, are included.Las colecciones científicas del Museo Nacional de Ciencias Naturales (MNCN-CSIC, Madrid albergan un extenso material entomológico recogido en Marruecos a principios del siglo XX por el insigne naturalista M. Mart

  15. Ectoparasitos de tilápia do Nilo (Oreochromis niloticus, das linhagens Chitralada e GIFT, em diferentes densidades e alimentadas com dois níveis de proteína - DOI: 10.4025/actascianimsci.v29i4.1015 Ectoparasites in Nile tilapia (Oreochromis niloticus from Chitralada and GIFT strains, in different densities, fed with two protein levels - DOI: 10.4025/actascianimsci.v29i4.1015

    Directory of Open Access Journals (Sweden)

    Maria de los Angeles Peres Lizama

    2008-03-01

    Full Text Available Foi verificada a infestação por ectoparasitos em tilápia do Nilo (Oreochromis niloticus, nas linhagens Chitralada e GIFT, em tanques e viveiros, utilizando-se ração com dois níveis de proteína. Durante o ensaio, foram analisados a temperatura, o pH, o oxigênio dissolvido e a condutividade elétrica. Foram realizadas amostras de raspados de tegumento e brânquias de machos pós-revertidos, em duas fases do experimento. A primeira, em 240 alevinos provenientes de 18 caixas de fibra de vidro de 500 L em três densidades. A prevalência total de parasitos na linhagem Chitralada (densidades de 30, 40 e 50 peixes m-3 foi 72,2, 83,3 e 59,5%, com predominância de Trichodina (38,9, 63,3 e 26,2%, respectivamente. Para a linhagem GIFT, nas mesmas densidades, foram observados 83,3, 73,3 e 80,9%, com maior predominância também de Trichodina (33,3, 73,3 e 45,2%, respectivamente. Na segunda fase, foram analisados 90 peixes de cada linhagem, de dois viveiros (140 m2 e duas dietas com 25 e 30% de proteína bruta. A prevalência total, para as linhagens Chitralada e GIFT com 25% PB foi 86,7 e 76,7%, respectivamente, e para 30% PB foi 60,0%, para ambas as linhagens. O nível de 30% PB, independentemente da linhagem, apresentou a menor prevalência parasitária.Ectoparasites infestation in Nile Tilapia (Oreochromis niloticus was observed in Chitralada and GIFT strains cultivated in cages and ponds, using rations with two protein levels. During the assay, temperature, pH, dissolved oxygen and electric conductivity were analyzed. Tegument scraping and gill samples from reverted males were evaluated, in two experiment phases. The first phase was carried out with 240 fingerlings from 18 500 L fiberglass boxes using three stocking densities. Total prevalence of parasites in the Chitralada strain (stocking density of 30, 40 and 50 fish m-3 was 72.2, 83.3 and 59.5%, with Trichodina predominance (38.9, 63.3 and 26.2%, respectively. For the GIFT strain, at the

  16. Efeitos do tratamento da Classe II divisão 1 em pacientes dolicofaciais tratados segundo a Terapia Bioprogressiva (AEB cervical e arco base inferior, com ênfase no controle vertical Treatment effects on Class II division 1 high angle patients treated according to the Bioprogressive therapy (cervical headgear and lower utility arch, with emphasis on vertical control

    Directory of Open Access Journals (Sweden)

    Viviane Santini Tamburús

    2011-06-01

    the use of cervical headgear (CHG in the maxillary arch, lower utility arch in the mandibular arch until normal occlusion of the molars was obtained and finished in accordance with Bioprogressive therapy, with a mean duration of 56 months. The values of FMA, SN.GoGn, ANB, Fg-S, S-FPm, maxillary length, mandibular length, posterior facial height (PFH, anterior facial height (AFH, facial height index (FHI, occlusal plane angle (OPA, palatal plane angle (PPA, total chin (TC, upper lip (UL and Z angle were evaluated. RESULTS: The results showed that treatment promoted stability of the mandibular, occlusal and palatal planes. Anteroposterior correction of the apical bases occurred, verified by the significant reduction in the variable ANB. The maxilla presented slight anterior displacement and increase in the anteroposterior dimension. The mandible presented improvement in its position in relation to the cranial base and its anteroposterior dimension increased significantly. The posterior and anterior facial heights remained in equilibrium, with no significant alteration in FHI. The tegumental profile presented significant improvement. CONCLUSION: The treatment performed produced correction of the apical basis with control of the horizontal planes and facial heights, and was effective for vertical control.

  17. Functionality of the plastron in adults of Neochetina eichhorniae Warner (Coleoptera, Curculionidae: aspects of the integument coating and submersion laboratory experiments Funcionalidade do plastrão em adultos de Neochetina eichhorniae Warner (Coleoptera, Curculionidae: aspectos do revestimento tegumentar e experimentos laboratoriais de submersão

    Directory of Open Access Journals (Sweden)

    Wesley Oliveira de Sousa

    2012-09-01

    Full Text Available The plastron theory was tested in adults of Neochetina eichhorniae Warner, 1970, through the analysis of the structure that coats these insects' integument and also through submersion laboratorial experiments. The tegument processes were recognized in three types: agglutinated scales with large perforations, plumose scales of varied sizes and shapes, and hairs. The experiments were carried out on 264 adult individuals which were kept submerged at different time intervals (n = 11 and in two types of treatment, natural non-aerated water and previously boiled water, with four repetitions for each treatment. The tests showed a maximum mortality after 24 hours of immersion in the previously boiled water treatment. The survival of the adults was negative and significantly correlated with the types of treatment employed and within the different time intervals. The values of oxygen dissolved in water (mg/l differed significantly within the types of treatment employed. They were positively correlated with the survival of the adults in the two types of treatment, although more markedly in the treatment with previously boiled water. The mortality of adults after 24 hours of submersion in the treatment with previously boiled water may be associated with the physical-chemical conditions of the non-tested water in this study, such as low surface tension and concentration of solutes. These results suggest plastron functionality in the adults of this species.A teoria plastrão foi testada em adultos de Neochetina eichhorniae Warner, 1970, por meio da análise da estrutura que reveste o tegumento destes insetos e em experimentos laboratoriais de submersão. Os processos tegumentares foram reconhecidos em três tipos: escamas aglutinadas e com perfurações largas; escamas plumosas de tamanhos e formas variadas; e pêlos. Os experimentos realizados com 264 indivíduos adultos os quais permaneciam submersos por diferentes intervalos de tempo (n = 11 e em dois

  18. Effect of the sun on visible clinical signs of aging in Caucasian skin

    Directory of Open Access Journals (Sweden)

    Flament F

    2013-09-01

    Full Text Available Frederic Flament,1 Roland Bazin,2 Sabine Laquieze,3 Virginie Rubert,1 Elisa Simonpietri,4 Bertrand Piot1 1Department of Applied Research and Development, L'Oreal Research and Innovation, Paris, France; 2RB Consult, Bievres, France; 3Private Dermatology Consultancy Practice, Montpellier, France; 4BIOTHERM International, Levallois-Perret, France Objectives: Aging signs can be classified into four main categories: wrinkles/texture, lack of firmness of cutaneous tissues (ptosis, vascular disorders, and pigmentation heterogeneities. During a lifetime, skin will change in appearance and structure not only because of chronological and intrinsic processes but also due to several external factors such as gravity, sun and ultraviolet exposure, and high levels of pollution; or lifestyle factors that have important and obvious effects on skin aging, such as diet, tobacco, illness, or stress. The effect of these external factors leads to progressive degradations of tegument that appear with different kinetics. The aim of this study was to clinically quantify the effect of sun exposure on facial aging in terms of the appearance of new specific signs or in terms of increasing the classical signs of aging. Materials and methods: This study was carried out on 298 Caucasian women from 30 years to 78 years old. The participants were divided into two groups according to their sun exposure history: 157 women were characterized as sun-seeking, and the other 141 were classified as sun-phobic. This division was made possible by dermatologist grading of heliodermal status on the basis of several observations of classic criteria: wrinkles, sagging, pigmentation heterogeneities, vascular disorders, elastosis, and so on. This work was an opportunity to complete clinical photographic tools by adding in our portfolio new scales for signs observed in the two groups. Thus, 22 clinical parameters were investigated by a panel of twelve trained experts to characterize each woman

  19. Wildfire Ash: Chemical Composition, Ash-Soil Interactions and Environmental Impacts

    Science.gov (United States)

    Brook, Anna; Hamzi, Seham; Wittenberg, Lea

    2015-04-01

    S., Morag H., 2013. The effect of wildfires on vegetation cover and dune activity in Australia's desert dunes: a multisensor analysis International Journal of Wildland Fire, vol. 21 (4), pp. 459-475. Lugassi R., Ben-Dor E., Eshel G., 2013. Reflectance spectroscopy of soils post-heating'Assessing thermal alterations in soil minerals. Geoderma, vol. 231, pp. 268-279. Pereira P., Úbeda X., Martin D., Mataix-Solera J., Guerrero C. 2011. Effects of a low prescribed fire in ash water soluble elements in a Cork Oak (Quercus suber) forest located in Northeast of Iberian Peninsula, Environmental Research, vol. 111(2), pp. 237-247. Shakesby R.A., 2011. Post-wildfire soil erosion in the Mediterranean: Review and future research directions Earth Science Reviews, vol. 105, pp. 71-100. Woods, S.W., Balfour, V.N. 2010. The effects of soil texture and ash thickness on the post-fire hydrological response from ash-covered soils, Journal of Hydrology, vol. 393, pp. 274-286.

  20. The impact of agriculture terraces on soil organic matter, aggregate stability, water repellency and bulk density. A study in abandoned and active farms in the Sierra de Enguera, Eastern Spain.

    Science.gov (United States)

    Cerdà, Artemi; Burguet, Maria; Keesstra, Saskia; Prosdocimi, Massimo; Di Prima, Simone; Brevik, Erik; Novara, Agata; Jordan, Antonio; Tarolli, Paolo

    2016-04-01

    results shows that the abandoned terrace is developing soils with more organic matter (7.34 % in average) than the control plot (5.37 %), with lower soil bulk density (1.01 g/cm3 against 1.05 g/cm3), higher WDPT (54 seconds against 42 seconds) and more stable aggregates (87 against 76 %). On the contrary, the active terrace shown soils with low more organic matter (2.05% in average) than the control plot non-terraced (5.39 %), with higher soil bulk density (1.12 g/cm3 against 1.06 g/cm3), lower WDPT (2.54 seconds against 43 seconds) and unstable aggregates (39 % surviving aggregates against 74 %). This results shown that terraces when abandoned are developing soils rich in organic matter, high aggregate stability, water repellent and low bulk density, but when active, the ploughing results in soils more degraded than the ones developed nearby. Acknowledgements The research leading to these results has received funding from the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement n° 603498 (RECARE project). Cerdà, A. The influence of aspect and vegetation on seasonal changes in erosion under rainfall simulation on a clay soil in Spain (1998) Canadian Journal of Soil Science, 78 (2), pp. 321-330. Cerdà, A., Lavee, H., Romero-Díaz, A., Hooke, J., Montanarella, L. 2010. Soil erosion and degradation in mediterranean type ecosystems. Land Degradation and Development, 21 (2), pp. 71-74..DOI: 10.1002/ldr.968 Gao Y., Dang X., Yu Y., Li Y., Liu Y., Wang J. Effects of Tillage Methods on Soil Carbon and Wind Erosion. (2015) Land Degradation and Development, . Article in Press. DOI: 10. 1002/ldr. 2404 Keesstra, S.D., 2007. Impact of natural reforestation on floodplain sedimentation in the Dragonja basin, SW Slovenia. Earth Surface Processes and Landforms, 32(1): 49-65. DOI: 10.1002/esp.1360 Li X. H., Yang J., Zhao C. Y., Wang B. Runoff and sediment from orchard terraces in southeastern China. (2014) Land Degradation and Development, 25 (2), pp. 184

  1. Crohn's Disease: current state of biological therapy Doença de Crohn: estado atual da terapia biológica

    Directory of Open Access Journals (Sweden)

    Júlio César Monteiro Santos Júnior

    2011-12-01

    Full Text Available The inflammatory bowel diseases (IBD are defined as nonspecific chronic intestinal inflammations with possible systemic involvement. IBD have unknown etiology. The inflammatory process is complex and heterogeneous, both as to the characterization of the disease that affects the digestive tract, without an intelligible pattern of revelation and balance, and in its different systemic damages when including the extensive and severe extraintestinal symptoms. Aparently, the natural history of the disease is irregular in relation to the offending agent system and the attacked system, both in the intestinal and extraintestinal teguments. Isolated aspects showing irregularity in this balance gives us the notion that IBD, especially Crohn's disease, can be caused by the stimulation of an immune response caused by damaging agents (intestinal bacteria, but mediated by inadequate genetic factors, whose expressions determine different individual susceptibilities. These observations have been shown in genetic studies that emphasize the importance of pathological interaction between host and bacteria subsidized by a genomic region that contains genes producing proteins (NOD2 - nucleotide-binding oligomerization domain containing 2 participating in an enhanced defense response by the tissue. Increased numbers and the activation of these cells in the intestinal mucosa elevate local levels of tumor necrosis factor α (TNF-α, interleukin-1β, interferon-Γ, and cytokines of the interleukin-23-Th17 pathway. So, it can be assumed that the susceptibility, which is a result of genetic alterations, is connected to an exaggerated response in the pro-inflammatory phase because of a dysfunction in the intestinal immune system. The identification of tumor necrosis factor (TNF-α as the active element in the pro-inflammatory inadequate response gave rise to the heightened production of biological substances that could block TNF-α, at different levels, opening a large

  2. Ciclo reprodutivo do caranguejo violinista Uca rapax (Smith (Crustacea, Brachyura, Ocypodidae habitante de um estuário degradado em Paraty, Rio de Janeiro, Brasil Reproductive cicle of the fiddler crab Uca rapax (Smith (Crustacea, Brachyura, Ocypodidae at a degraded estuary in Paraty, Rio de Janeiro, Brazil

    Directory of Open Access Journals (Sweden)

    Daniela da S. Castiglioni

    2006-06-01

    cycles in a degraded mangrove area in Paraty, State of Rio de Janeiro. Two collectors captured the crabs monthly (from July 2001 to June 2002 during 15 minutes by procedure of capture per unit effort in low tide periods. Crabs were sexed, measured (carapace width - CW; 0.01 mm precision and females were checked for eggs. Crab' stages of gonad development of both sexes were determined by direct and macroscopic observation and molt stage was estimated by the hardness of the tegument. A total of 1,558 specimens were collected, being 801 males and 757 females (16 ovigerous females. Ovigerous females accounted only about 3% of the population, perhaps because females usually remain underground in closed burrows during the incubation, which can not be seen easily in the field. Crabs with mature gonads were found year-round, but most reproduction in U. rapax occurred during the warmer months of the year (spring-summer. The frequency of crabs in molt activity was lower (12.8% than intermolt crabs. Probably, U. rapax burrowed in this critic period that is the change of exoskeleton, minimizing the risks imposed by the occurrence of a soft skeleton. Despite of U. rapax be occurring in a properly degraded mangrove, its reproductive cycle seems to be not affected by the habitat condition, when it is compared with areas of the mangroves no impacted by human action. This fact is supported by the presence of potentially mature crabs year-round at the study area.

  3. Storage and methods for overcoming dormancy of the coral tree seedsArmazenamento e métodos para a superação da dormência de sementes de mulungu

    Directory of Open Access Journals (Sweden)

    Ana Claudia da Silva

    2013-03-01

    Full Text Available The coral tree (Erythrina velutina Willd. is an arboreal species that occurs in northeast Brazil and is used in popular medicine to fight cough, asthma, oral diseases, nervous breakdowns, muscle aches and fevers. The process of formation of seedlings of the species is difficult because of the occurrence of seed dormancy. The aim of this study was to evaluate the effect of different methods for overcoming dormancy in seeds of the coral tree stored in different conditions and periods. The experimental design was completely randomized in factorial scheme 5 x 2 (methods for overcoming dormancy and storage types, in four replicates of 25 seeds. The methods used were (S1 control, (S2 scarification opposite the micropyle; (S3 by the scarification micropyle; (S4 seeds immersed in water at 80 °C for five minutes and (S5 seeds immersed in water at 100 °C for two minutes. The storage types were for 25 months at cold chamber and for a month at the laboratory environment. The characteristics evaluated were emergence percentage (PE, time of emergence (TM, shoot length (CPA and root (CR, diameter (D, leaf number (NF, dry biomass of roots (MSPA and of shoots (MSR of seedlings. The emergence percentage was low in treatments S1, S4 and S5 and the seeds stored for one month were more affected when immersed in hot water. The highest values of CPA, CR, MSPA and NF treatments were observed in S2 and S3, which occurred in the lowest average time of emergence. The tegument dormancy of seeds of Erythrina velutina is overcome by scarification with sandpaper on the end opposite the micropyle or next to it. The seed dormancy is not affected by the types of storage and is not overcome by immersion in hot water. O mulungu (Erythrina velutina Willd. é uma espécie arbórea que ocorre no Nordeste brasileiro utilizada na medicina popular no combate a tosse, asma, afecções bucais, crises nervosas, dores musculares e febres. O processo de formação de mudas da espécie

  4. Métodos para acelerar a germinação de sementes de Bacuri (Platonia insignis Mart. Methods to accelerate the germination of Bacuri seeds (Platonia insignis Mart.

    Directory of Open Access Journals (Sweden)

    FRANCISCO DAS CHAGAS OLIVEIRA

    2002-04-01

    Full Text Available Este trabalho teve o objetivo de avaliar métodos para acelerar a germinação de sementes de bacuri (Platonia insignis Mart., e foi conduzido no laboratório de Fisiologia Vegetal e na Câmara de Nebulização da Embrapa Meio-Norte, Teresina-PI. O delineamento utilizado foi o de blocos ao acaso, constituído por dez tratamentos e quatro repetições, sendo a unidade experimental constituída por dez sementes. Foram testados os seguintes tratamentos: testemunha (T1; remoção do tegumento da semente (T2; remoção do meristema fundamental cortical por meio de cortes em planos perpendiculares ao plano dorsal/ventral, nos dois lados da semente, sem atingir o meristema fundamental medular (T3; T3 mais a remoção do meristema fundamental cortical por meio de cortes em planos paralelos ao plano dorsal/ventral, na região dorsal, sem atingir o meristema fundamental medular (T4; T3 e T4 mantidos em água a 40ºC por vinte minutos (T5 e T6; T3 e T4 mantidos em etanol 80% por cinco minutos (T7 e T8; T3 e T4 mantidos em acetona 80% por cinco minutos (T9 e T10. As variáveis estudadas foram a percentagem de emergência da radícula aos 14; 21; 28 e 35 dias após a semeadura e o índice de velocidade de emergência da radícula (IVE, no 35º dia após a semeadura. As sementes submetidas aos tratamentos T2, T3, T4, T5 e T6 apresentaram 72,5%, 65,0%, 72,5%, 52,5% e 67,5% de emergência, respectivamente, aos 14 dias, sendo superiores (pThe objective of this study was to evaluate methods to accelerate the germination of bacuri seeds (Platonia insignis Mart.. The experiment was conducted in the laboratory of Plant Physiology and in the greenhouse of Embrapa Meio-Norte (Teresina-PI, in a randomized block design, constituted by ten treatments and four repetitions, being the experimental unit constituted by ten seeds. The following treatments were tested: control (T1; removal of the seed tegument (T2; removal of the fundamental meristem, through cuts in

  5. Colonização e lesão em fêmeas ingurgitadas do carrapato Rhipicephalus sanguineus causadas pelo fungo Metarhizium anisopliae Colonization and lesions on engorged female Rhipicephalus sanguineus, caused by Metarhizium anisopliae

    Directory of Open Access Journals (Sweden)

    Marcos Valerio Garcia

    2004-10-01

    . (SOROKIN, 1883], in to the Rhipicephalus sanguineus ticks (LATREILLE, 1806, as well as the lesions caused in the tissues of the acarus. The form of adherence and penetration of the fungus in the deep tissue was studied through scanning microscopy and the fungus action on the internal tissues was evaluated in conventional hystological sections. In order to evaluate these events, experimental infestations were conducted in 11 engorged females groups of the tick R. sanguineus, containing 12 engorged females in each. Aiming this goal the engorged females were submited to a bath in a conidial suspension in a concentration of 108 conidia/mL during three minutes under manual agitation. In case of control groups the bath was done only in the suspension vehicle. Ticks were processed for histological and scanning microscopy surveys at different intervals after the infection: 1 and 18h and one, two, three, four, five, six, seven, eight, nine and eleven days. It has been observed that the major part of the conidians have germinated within 18h after the inoculation and that the fungus the penetrated the acarus body, trough it´s tegument 48h after the infection. After the penetration the fungus colonized the tick’s body in a diffuse manner, without any preference for a specific tissue. Among the lesions caused by the fungus, rupture of the tick’s intestinal wall and the dispersion of the fluid into the hemocel were the most outstanding features. Death of the host occurred 96h pos-infection and the sporulation of the pathogen over the dead acarus began 120 to 144h pos-infection. This work aims to contribute to the establishement of techniques for biological control of the ticks by fungus as an ecological alternative for acaricides.

  6. [Traumatic knee dislocation with popliteal vascular disruption: retrospective study of 14 cases].

    Science.gov (United States)

    Bonnevialle, P; Chaufour, X; Loustau, O; Mansat, P; Pidhorz, L; Mansat, M

    2006-12-01

    . On average, limb revascularization was achieved after 10.07 hours ischemia. Intravenous heparin was instituted for 810 days followed by low-molecular-weight heparin. The dislocation was stabilized by a femorotibial fixator in nine patients and a cruropedious cast in five. An incision was made in the anterolateral and posterior leg compartments in twelve patients. A revision procedure was necessary on day one in one patient because of recurrent ischemia; a second bypass using an autologous venous graft was successful. One other 75-year-old patient also presented recurrent ischemia on day five; the bypass was reconstructed but the patient died from multiple injuries. Seven thin skin grafts were used to cover the aponeurotomy surfaces. Mean duration of the external fixator was 3.4 months. The five patients treated with a plaster case were immobilized for 2.7 months on average. Ligament repair was performed in three patients (one lateral reconstruction and one double reconstruction of the central pivot for the two others). A total prosthesis with a rotating hinge was implanted in two patients aged 67 and 74 years after removal of the external fixator at six and seven months. Failure of the ligament repair also led to arthroplasty in a third patient. Blood supply to the lower limb was successfully restored as proven by the renewed coloration of the teguments and-or presence of distal pulses in 13 patients. Transient acute renal failure required dialysis in one patient. Four patients developed pin track discharges and there was one case of septic arthritis of the knee joint which was cured after arthrotomy for wash-out and adapted antibiotics. Outcome was assessed a minimum 18 months follow-up (average 22 months) for the 13 survivors. The three sciatic palsies recovered partially at five and six months in the tibial territory but with persistent paralysis in the territory of the common peroneal nerve. The nine cases of common peroneal nerve palsy noted initially regressed