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Sample records for gondii cyclophilin 18-mediated

  1. CYCLOPHILIN A: STRUCTURE AND FUNCTIONS

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    A. A. Kalinina

    2017-01-01

    Full Text Available Cyclophilins belong to a large family of ancient conservative proteins with peptidyl-prolyl-cis-trans isomerase activity. The main member of this family – cyclophilin A – was discovered as an intracellular ligand for cyclosporine A. Further investigations revealed a wide range of functions of cyclophilin A. Cyclophilin A is involved in T-cell signaling, it takes part in folding, assembly and intracellular transport of proteins, as well as acts as an antioxidant. Different cell types secrete cyclophilin A under infection or oxidative stress. Cyclophilin A is one of the main factors involved in inflammation and pathogenesis of autoimmune, cardiovascular and other diseases. This protein is thought to take part in tumor progression. In this review we describe the structure of cyclophilin A and its main known functions in health and disease.

  2. 45 CFR 16.18 - Mediation.

    Science.gov (United States)

    2010-10-01

    ... 45 Public Welfare 1 2010-10-01 2010-10-01 false Mediation. 16.18 Section 16.18 Public Welfare... BOARD § 16.18 Mediation. (a) In cases pending before the Board. If the Board decides that mediation... mediation techniques and will provide or assist in selecting a mediator. The mediator may take any steps...

  3. Recombinant Cyclophilins Lack Nuclease Activity

    OpenAIRE

    Manteca, Angel; Sanchez, Jesus

    2004-01-01

    Several single-domain prokaryotic and eukaryotic cyclophilins have been identified as also being unspecific nucleases with a role in DNA degradation during the lytic processes that accompany bacterial cell death and eukaryotic apoptosis. Evidence is provided here that the supposed nuclease activity of human and bacterial recombinant cyclophilins is due to contamination of the proteins by the host Escherichia coli endonuclease and is not an intrinsic property of these proteins.

  4. Isoform-specific inhibition of cyclophilins.

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    Daum, Sebastian; Schumann, Michael; Mathea, Sebastian; Aumüller, Tobias; Balsley, Molly A; Constant, Stephanie L; de Lacroix, Boris Féaux; Kruska, Fabian; Braun, Manfred; Schiene-Fischer, Cordelia

    2009-07-07

    Cyclophilins belong to the enzyme class of peptidyl prolyl cis-trans isomerases which catalyze the cis-trans isomerization of prolyl bonds in peptides and proteins in different folding states. Cyclophilins have been shown to be involved in a multitude of cellular functions like cell growth, proliferation, and motility. Among the 20 human cyclophilin isoenzymes, the two most abundant members of the cyclophilin family, CypA and CypB, exhibit specific cellular functions in several inflammatory diseases, cancer development, and HCV replication. A small-molecule inhibitor on the basis of aryl 1-indanylketones has now been shown to discriminate between CypA and CypB in vitro. CypA binding of this inhibitor has been characterized by fluorescence anisotropy- and isothermal titration calorimetry-based cyclosporin competition assays. Inhibition of CypA- but not CypB-mediated chemotaxis of mouse CD4(+) T cells by the inhibitor provided biological proof of discrimination in vivo.

  5. Toxoplasma gondii and schizophrenia

    OpenAIRE

    Mokhtari Mohammadreza; Mokhtari Mojgan

    2006-01-01

    Recent epidemiologic studies indicate that infectious agents may contribute to some cases of schizophrenia. In animals, infection with Toxoplasma gondii can alter behavior and neurotransmitter function. In humans, acute infection with T. gondii can produce psychotic symptoms similar to those displayed by persons with schizophrenia. Since 1953, a total of 19 studies of T. gondii antibodies in persons with schizophrenia and other severe psychiatric disorders and in controls have been reported; ...

  6. Suppression of Coronavirus Replication by Cyclophilin Inhibitors

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    Takashi Sasaki

    2013-05-01

    Full Text Available Coronaviruses infect a variety of mammalian and avian species and cause serious diseases in humans, cats, mice, and birds in the form of severe acute respiratory syndrome (SARS, feline infectious peritonitis (FIP, mouse hepatitis, and avian infectious bronchitis, respectively. No effective vaccine or treatment has been developed for SARS-coronavirus or FIP virus, both of which cause lethal diseases. It has been reported that a cyclophilin inhibitor, cyclosporin A (CsA, could inhibit the replication of coronaviruses. CsA is a well-known immunosuppressive drug that binds to cellular cyclophilins to inhibit calcineurin, a calcium-calmodulin-activated serine/threonine-specific phosphatase. The inhibition of calcineurin blocks the translocation of nuclear factor of activated T cells from the cytosol into the nucleus, thus preventing the transcription of genes encoding cytokines such as interleukin-2. Cyclophilins are peptidyl-prolyl isomerases with physiological functions that have been described for many years to include chaperone and foldase activities. Also, many viruses require cyclophilins for replication; these include human immunodeficiency virus, vesicular stomatitis virus, and hepatitis C virus. However, the molecular mechanisms leading to the suppression of viral replication differ for different viruses. This review describes the suppressive effects of CsA on coronavirus replication.

  7. Taxonomy Icon Data: Toxoplasma gondii [Taxonomy Icon

    Lifescience Database Archive (English)

    Full Text Available Toxoplasma gondii Toxoplasma gondii Toxoplasma_gondii_L.png Toxoplasma_gondii_NL.png Toxoplasma..._gondii_S.png Toxoplasma_gondii_NS.png http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Toxoplasma...+gondii&t=L http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Toxoplasma+gondii&t=NL http://biosciencedbc.j...p/taxonomy_icon/icon.cgi?i=Toxoplasma+gondii&t=S http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Toxoplas...ma+gondii&t=NS http://togodb.biosciencedbc.jp/togodb/view/taxonomy_icon_comment_en?species_id=113 ...

  8. The ability of multimerized cyclophilin A to restrict retrovirus infection

    International Nuclear Information System (INIS)

    Javanbakht, Hassan; Diaz-Griffero, Felipe; Yuan Wen; Yeung, Darwin F.; Li Xing; Song Byeongwoon; Sodroski, Joseph

    2007-01-01

    In owl monkeys, the typical retroviral restriction factor of primates, TRIM5α, is replaced by TRIMCyp. TRIMCyp consists of the TRIM5 RING, B-box 2 and coiled-coil domains, as well as the intervening linker regions, fused with cyclophilin A. TRIMCyp restricts infection of retroviruses, such as human immunodeficiency virus (HIV-1) and feline immunodeficiency virus (FIV), with capsids that can bind cyclophilin A. The TRIM5 coiled coil promotes the trimerization of TRIMCyp. Here we show that cyclophilin A that is oligomeric as a result of fusion with a heterologous multimer exhibits substantial antiretroviral activity. The addition of the TRIM5 RING, B-box 2 and Linker 2 to oligomeric cyclophilin A generated a protein with antiretroviral activity approaching that of wild-type TRIMCyp. Multimerization increased the binding of cyclophilin A to the HIV-1 capsid, promoting accelerated uncoating of the capsid and restriction of infection

  9. Cyclophilin A interacts with diverse lentiviral capsids

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    Emerman Michael

    2006-10-01

    Full Text Available Abstract Background The capsid (CA protein of HIV-1 binds with high affinity to the host protein cyclophilin A (CypA. This binding positively affects some early stage of the viral life-cycle because prevention of binding either by drugs that occupy that active site of cyclophilin A, by mutation in HIV-1 CA, or RNAi that knocks down intracellular CypA level diminishes viral infectivity. The closely related lentivirus, SIVcpz also binds CypA, but it was thought that this interaction was limited to the HIV-1/SIVcpz lineage because other retroviruses failed to interact with CypA in a yeast two-hybrid assay. Results We find that diverse lentiviruses, FIV and SIVagmTAN also bind to CypA. Mutagenesis of FIV CA showed that an amino acid that is in a homologous position to the proline at amino acid 90 of HIV-1 CA is essential for FIV interactions with CypA. Conclusion These results demonstrate that CypA binding to lentiviruses is more widespread than previously thought and suggest that this interaction is evolutionarily important for lentiviral infection.

  10. Expression and Significance of Cyclophilin J in Primary Gastric Adenocarcinoma.

    Science.gov (United States)

    Gong, Zhaohua; Mu, Yuling; Chen, Jian; Chu, Hongjin; Lian, Peiwen; Wang, Congcong; Wang, Jiahui; Jiang, Lixin

    2017-08-01

    Biomarkers are essential in early diagnosis and understanding of the molecular mechanism of human cancer. The expression of cyclophilin J, a novel member of the cyclophilin family, was investigated in primary gastric adenocarcinoma. Western blot analysis was carried out on 36 paired tumor and normal tissue samples; immunohistochemical analysis was carried out on 120 gastric carcinoma tissues and normal adjacent tissue. Cyclophilin J protein was overexpressed in 72.2% of gastric carcinoma tissues compared to adjacent normal tissues. Immunohistochemical analysis revealed that cyclophilin J was overexpressed in 49.2% (59/120) and 23.3% (28/120) of gastric carcinoma tissues and adjacent tissues, respectively (pJ was associated with the degree of differentiation, but not with lymph node metastasis, gender or depth of tumor infiltration. The overall survival of patients showed no association with the overexpression of cyclophilin J protein. Cyclophilin J expression was up-regulated in gastric carcinoma compared to normal gastric tissues. However, in order to confirm its association with the survival of patients with gastric cancer, more cases need to be studied. Copyright© 2017, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.

  11. Cyclophilin B enhances HIV-1 infection

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    DeBoer, Jason; Madson, Christian J. [Department of Medical Microbiology and Immunology, Creighton University, Omaha, NE (United States); Belshan, Michael, E-mail: michaelbelshan@creighton.edu [Department of Medical Microbiology and Immunology, Creighton University, Omaha, NE (United States); The Nebraska Center for Virology, University of Nebraska, Lincoln, NE (United States)

    2016-02-15

    Cyclophilin B (CypB) is a member of the immunophilin family and intracellular chaperone. It predominantly localizes to the ER, but also contains a nuclear localization signal and is secreted from cells. CypB has been shown to interact with the Gag protein of human immunodeficiency type 1 (HIV-1). Several proteomic and genetic studies identified it as a potential factor involved in HIV replication. Herein, we show that over-expression of CypB enhances HIV infection by increasing nuclear import of viral DNA. This enhancement was unaffected by cyclosporine treatment and requires the N-terminus of the protein. The N-terminus contains an ER leader sequence, putative nuclear localization signal, and is required for secretion. Deletion of the N-terminus resulted in mislocalization from the ER and suppression of HIV infection. Passive transfer experiments showed that secreted CypB did not impact HIV infection. Combined, these experiments show that intracellular CypB modulates a pathway of HIV nuclear import. - Highlights: • CypB has been identified in several proteomic studies of HIV-1 infection. • CypB expression is upregulated in activated and infected T-cells. • Over-expression of CypB enhances HIV nuclear import and infection. • The N-terminus of CypB is necessary for these effects.

  12. Cyclophilin B enhances HIV-1 infection

    International Nuclear Information System (INIS)

    DeBoer, Jason; Madson, Christian J.; Belshan, Michael

    2016-01-01

    Cyclophilin B (CypB) is a member of the immunophilin family and intracellular chaperone. It predominantly localizes to the ER, but also contains a nuclear localization signal and is secreted from cells. CypB has been shown to interact with the Gag protein of human immunodeficiency type 1 (HIV-1). Several proteomic and genetic studies identified it as a potential factor involved in HIV replication. Herein, we show that over-expression of CypB enhances HIV infection by increasing nuclear import of viral DNA. This enhancement was unaffected by cyclosporine treatment and requires the N-terminus of the protein. The N-terminus contains an ER leader sequence, putative nuclear localization signal, and is required for secretion. Deletion of the N-terminus resulted in mislocalization from the ER and suppression of HIV infection. Passive transfer experiments showed that secreted CypB did not impact HIV infection. Combined, these experiments show that intracellular CypB modulates a pathway of HIV nuclear import. - Highlights: • CypB has been identified in several proteomic studies of HIV-1 infection. • CypB expression is upregulated in activated and infected T-cells. • Over-expression of CypB enhances HIV nuclear import and infection. • The N-terminus of CypB is necessary for these effects.

  13. Cyclophilin B enhances HIV-1 infection.

    Science.gov (United States)

    DeBoer, Jason; Madson, Christian J; Belshan, Michael

    2016-02-01

    Cyclophilin B (CypB) is a member of the immunophilin family and intracellular chaperone. It predominantly localizes to the ER, but also contains a nuclear localization signal and is secreted from cells. CypB has been shown to interact with the Gag protein of human immunodeficiency type 1 (HIV-1). Several proteomic and genetic studies identified it as a potential factor involved in HIV replication. Herein, we show that over-expression of CypB enhances HIV infection by increasing nuclear import of viral DNA. This enhancement was unaffected by cyclosporine treatment and requires the N-terminus of the protein. The N-terminus contains an ER leader sequence, putative nuclear localization signal, and is required for secretion. Deletion of the N-terminus resulted in mislocalization from the ER and suppression of HIV infection. Passive transfer experiments showed that secreted CypB did not impact HIV infection. Combined, these experiments show that intracellular CypB modulates a pathway of HIV nuclear import. Copyright © 2015 Elsevier Inc. All rights reserved.

  14. Role of cyclophilins in somatolactogenic action.

    Science.gov (United States)

    Rycyzyn, M A; Clevenger, C V

    2000-01-01

    Prolactin (PRL) and growth hormone (GH) are members of the somatolactogenic hormone family, the pleiotropic actions of which are necessary for vertebrate growth and mammary differentiation. The basis for the specific function of these hormones has remained uncertain; however, their action is associated with internalization and translocation into the nucleus. A yeast two-hybrid screen identified an interaction between PRL and cyclophilin B (CypB), a peptidyl prolyl isomerase (PPI) found in the endoplasmic reticulum (ER), extracellular space, and nucleus. The interaction between CypB and PRL/GH was confirmed in vitro and in vivo through the use of recombinant proteins and coimmunoprecipitation studies. The exogenous addition of CypB potentiated the proliferation of PRL- and GH-dependent cell lines 18- and 40-fold, respectively. The potentiation of PRL action by CypB was accompanied by a dramatic increase in the nuclear retrotranslocation of PRL. Immunogold electron microscopy has revealed this retrotransport to occur via a vesicular pathway. A CypB mutant, termed CypB-NT, was generated that lacked the putative wild-type N-terminal nuclear localization sequence. Although CypB-NT demonstrated levels of PRL binding and PPI activity equivalent to wild-type CypB, it was incapable of mediating the nuclear retrotranslocation of PRL or enhancing PRL-driven proliferation. These studies reveal CypB as an important chaperone facilitating the nuclear retrotransport and action of the somatolactogenic hormone family.

  15. Toxoplasma gondii and Epilepsy.

    Science.gov (United States)

    Ayaz, Erol; Türkoğlu, Şule Aydın; Orallar, Hayriye

    2016-06-01

    Toxoplasma gondii is a zoonotic parasite can be seen in all the vital organ; in the acute phase, it can be found in the blood, cerebrospinal fluid, semen, tears, saliva, urine, and in almost all body fluids. Transplasental infection can lead to fetal damage and miscarriage. Its last hosts are felines and intermediate hosts are all mammals, including humans. People infected by the ingestion of meat containing cysts in undercooked or raw, are thrown oocysts with cat felines By taking in water and food, from mother to fetus transplacental way, the infected organ transplantation, blood transfusion, laboratory accidents and kaprofaj transmitted by mechanical vectors of the invertebrates. Suppression of the immune system is being transformed to the shape and texture of the cysts with bradyzoite. The parasite settles in the cells of the tissue cysts and causes change in the cellular mechanisms, such as cytokinin task. Depending on changes and type of neurotransmitter (GABA, glutamate, serotonin, dopamine) levels in CSF in ions (Ca, K, Cl, Mg), it is believed that there is a change in their concentration. In this review, literature about the relationship between T. gondii and epilepsy and epileptiform activity the importance of parasites, which settle in the brain, will be highlighted.

  16. Veterinary vaccines against Toxoplasma gondii

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    Elisabeth A Innes

    2009-03-01

    Full Text Available Toxoplasma gondii has a very wide intermediate host range and is thought to be able to infect all warm blooded animals. The parasite causes a spectrum of different diseases and clinical symptoms within the intermediate hosts and following infection most animals develop adaptive humoral and cell-mediated immune responses. The development of protective immunity to T. gondii following natural infection in many host species has led researchers to look at vaccination as a strategy to control disease, parasite multiplication and establishment in animal hosts. A range of different veterinary vaccines are required to help control T. gondii infection which include vaccines to prevent congenital toxoplasmosis, reduce or eliminate tissue cysts in meat producing animals and to prevent oocyst shedding in cats. In this paper we will discuss some of the history, challenges and progress in the development of veterinary vaccines against T. gondii.

  17. Molecular cloning and characterization of Aspergillus nidulans cyclophilin B.

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    Joseph, J D; Heitman, J; Means, A R

    1999-06-01

    Cyclophilins are an evolutionarily conserved family of proteins which serve as the intracellular receptors for the immunosuppressive drug cyclosporin A. Here we report the characterization of the first cyclophilin cloned from the filamentous fungus Aspergillus nidulans (CYPB). Sequence analysis of the cypB gene predicts an encoded protein with highest homology to the murine cyclophilin B protein. The sequence similarity includes an N-terminal sequence predicted to target the protein to the endoplasmic reticulum (ER) as well as a C-terminal sequence predicted to retain the mature protein in the ER. The bacterially expressed hexa-histidine tagged protein displays peptidyl-prolyl isomerase activity which is inhibited by cyclosporin A. In the presence of cyclosporin A, the expressed protein also inhibits purified calcineurin. When the endogenous cypB gene was disrupted and placed under the control of the regulatable alcohol dehydrogenase promoter, the strain demonstrated no detectable growth phenotype under conditions which induce or repress cypB transcription. Induction or repression of the cypB gene also did not effect sensitivity of A. nidulans to cyclosporin A. cypB mRNA levels were significantly elevated under severe heat shock conditions, indicating a possible role for the A. nidulans cyclophilin B protein during growth in high stress environments. Copyright 1999 Academic Press.

  18. Cyclophilin and Viruses: Cyclophilin as a Cofactor for Viral Infection and Possible Anti-Viral Target

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    Koichi Watashi

    2007-01-01

    Full Text Available Cyclophilin (CyP is a peptidyl prolyl cis/trans isomerase, catalyzing the cis-trans isomerization of proline residues in proteins. CyP plays key roles in several different aspects of cellular physiology including the immune response, transcription, mitochondrial function, cell death, and chemotaxis. In addition to these cellular events, a number of reports demonstrated that CyP plays a critical role in the life cycle of viruses, especially human immunodeficiency virus (HIV and hepatitis C virus (HCV. These two viruses are significant causes of morbidity and mortality worldwide, but current therapies are often insufficient. CyP may provide a novel therapeutic target for the management and/or cure of these diseases, in particular HCV.

  19. Human cyclophilin B: A second cyclophilin gene encodes a peptidyl-prolyl isomerase with a signal sequence

    International Nuclear Information System (INIS)

    Price, E.R.; Zydowsky, L.D.; Jin, Mingjie; Baker, C.H.; McKeon, F.D.; Walsh, C.T.

    1991-01-01

    The authors report the cloning and characterization of a cDNA encoding a second human cyclosporin A-binding protein (hCyPB). Homology analyses reveal that hCyPB is a member of the cyclophilin B (CyPB) family, which includes yeast CyPB, Drosophila nina A, and rat cyclophilin-like protein. This family is distinguished from the cyclophilin A (CyPA) family by the presence of endoplasmic reticulum (ER)-directed signal sequences. hCyPB has a hydrophobic leader sequence not found in hCyPA, and its first 25 amino acids are removed upon expression in Escherichia coli. Moreover, they show that hCyPB is a peptidyl-prolyl cis-trans isomerase which can be inhibited by cyclosporin A. These observations suggest that other members of the CyPB family will have similar enzymatic properties. Sequence comparisons of the CyPB proteins show a central, 165-amino acid peptidyl-prolyl isomerase and cyclosprorin A-binding domain, flanked by variable N-terminal and C-terminal domains. These two variable regions may impart compartmental specificity and regulation to this family of cyclophilin proteins containing the conserved core domain. Northern blot analyses show that hCyPB mRNA is expressed in the Jurkat T-cell line, consistent with its possible target role in cyclosporin A-mediated immunosuppression

  20. Solution characterization of the extracellular region of CD147 and its interaction with its enzyme ligand cyclophilin-A

    Energy Technology Data Exchange (ETDEWEB)

    Schlegel, Jennifer; Redzic, Jasmina S.; Porter, Christopher; Yurchenko, Vyacheslav; Bukrinsky, Michael; Labeikovsky, Wladimir; Armstrong, Geoffrey S.; Zhang, Fengli; Isern, Nancy G.; Degregori, James; Hodges, Robert; Eisenmesser, Elan Z.

    2009-08-21

    The CD147 receptor plays an integral role in numerous diseases by stimulating the expression of several protein families and serving as the receptor for extracellular cyclophilins, however, neither CD147 nor its interactions with its cyclophilin ligands have been well characterized in solution. CD147 is a unique protein in that it can function both at the cell membrane and after being released from cells where it continues to retain activity. Thus, the CD147 receptor functions through at least two mechanisms that include both cyclophilin-independent and cyclophilin-dependent modes of action. In regard to CD147 cyclophilin-independent activity, CD147 homophilic interactions are thought to underlie its activity. In regard to CD147 cyclophilin-dependent activity, cyclophilin/CD147 interactions may represent a novel means of signaling since cyclophilins are also peptidyl-prolyl isomerases.

  1. CD147 is a signaling receptor for cyclophilin B.

    Science.gov (United States)

    Yurchenko, V; O'Connor, M; Dai, W W; Guo, H; Toole, B; Sherry, B; Bukrinsky, M

    2001-11-09

    Cyclophilins A and B (CyPA and CyPB) are cyclosporin A binding proteins that can be secreted in response to inflammatory stimuli. We recently identified CD147 as a cell-surface receptor for CyPA and demonstrated that CD147 is an essential component in the CyPA-initiated signaling cascade that culminates in ERK activation and chemotaxis. Here we demonstrate that CD147 also serves as a receptor for CyPB. CyPB induced Ca(2+) flux and chemotaxis of CD147-transfected, but not control, CHO cells, and the chemotactic response of primary human neutrophils to CyPB was blocked by antibodies to CD147. These results suggest that CD147 serves as a receptor for extracellular cyclophilins. Copyright 2001 Academic Press.

  2. Cyclophilin B facilitates the replication of Orf virus

    OpenAIRE

    Zhao, Kui; Li, Jida; He, Wenqi; Song, Deguang; Zhang, Ximu; Zhang, Di; Zhou, Yanlong; Gao, Feng

    2017-01-01

    Background Viruses interact with host cellular factors to construct a more favourable environment for their efficient replication. Expression of cyclophilin B (CypB), a cellular peptidyl-prolyl cis-trans isomerase (PPIase), was found to be significantly up-regulated. Recently, a number of studies have shown that CypB is important in the replication of several viruses, including Japanese encephalitis virus (JEV), hepatitis C virus (HCV) and human papillomavirus type 16 (HPV 16). However, the f...

  3. Feline coronavirus replication is affected by both cyclophilin A and cyclophilin B.

    Science.gov (United States)

    Tanaka, Yoshikazu; Sato, Yuka; Sasaki, Takashi

    2017-02-01

    Feline coronavirus (FCoV) causes the fatal disease feline infectious peritonitis, which is currently incurable by drug treatment, and no effective vaccines are available. Cyclosporin A (CsA), a cyclophilin (Cyp) inhibitor, inhibits the replication of FCoV in vitro and in vivo as well as the replication of human and animal coronaviruses. However, the mechanism underlying the regulation of coronavirus replication by CsA is unknown. In this study, we analysed the role of Cyps in FCoV replication using knockdown and knockout cells specific to Cyps. Inhibition of CypA and CypB reduced FCoV replication, with replication in knockout cells being much less than that in knockdown cells. Furthermore, the proteins expressed by CypA and CypB harbouring mutations in their respective predicted peptidyl-prolyl cis-transisomerase active sites, which also alter the affinities between Cyps and CsA, inhibited FCoV replication. These findings indicate that the peptidyl-prolyl cis-transisomerase active sites of Cyps might be required for FCoV replication.

  4. Seroprevalence of Toxoplasma gondii in Western Romania.

    Science.gov (United States)

    Olariu, Tudor Rares; Petrescu, Cristina; Darabus, Gheorghe; Lighezan, Rodica; Mazilu, Octavian

    2015-08-01

    Toxoplasma gondii is an obligate intracellular parasite that most commonly causes asymptomatic infection in immunocompetent hosts, but can have devastating consequences in congenitally infected infants and immunocompromised patients. We evaluated the seroprevalence of T. gondii in the general population in Western Romania. Sera from 304 individuals were analysed with the Pastorex Toxo test, which allows the simultaneous detection of T. gondii IgG and/or IgM antibodies. T. gondii antibodies were demonstrated in 197 individuals (64.8%) and the prevalence increased with age: 35.0% in those Romania.

  5. Seroprevalence of Toxoplasma gondii infection amongst residents ...

    African Journals Online (AJOL)

    Toxoplasmosis is a zoonotic disease, recognized as a serious public health problem worldwide. Toxoplasma gondii infection has become a major public health concern in recent years due to the ravaging HIV/AIDS pandemic. A serological survey was carried out in Tanga district of north-eastern Tanzania to assess T. gondii ...

  6. Toxoplasma gondii, Mental Health and Shizophrenia

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    Sibel Cevizci

    2013-04-01

    Full Text Available Protecting and promoting of mental health is one of the major application areas of public health. In particular, Toxoplasma gondii, which is a protozoal zoonosis common in Turkey, it is closely related to veterinary public health. In recent years, T.gondii can induce behavioral changes, may play a role in schizophrenia as an etiologic factor. Results of the recently performed studies shows that T.gondii may be a potential factor for some neuropathological changes in brain and suicide attemption. The purpose of this review is to present the data on recent epidemiology of T.gondii, mental health effects (changes in behavior, suicide, etc., the relationship between T.gondii and schizophrenia and offer some recommendations for protecting of public health. [TAF Prev Med Bull 2013; 12(2.000: 199-208

  7. Target cell cyclophilins facilitate human papillomavirus type 16 infection.

    Science.gov (United States)

    Bienkowska-Haba, Malgorzata; Patel, Hetalkumar D; Sapp, Martin

    2009-07-01

    Following attachment to primary receptor heparan sulfate proteoglycans (HSPG), human papillomavirus type 16 (HPV16) particles undergo conformational changes affecting the major and minor capsid proteins, L1 and L2, respectively. This results in exposure of the L2 N-terminus, transfer to uptake receptors, and infectious internalization. Here, we report that target cell cyclophilins, peptidyl-prolyl cis/trans isomerases, are required for efficient HPV16 infection. Cell surface cyclophilin B (CyPB) facilitates conformational changes in capsid proteins, resulting in exposure of the L2 N-terminus. Inhibition of CyPB blocked HPV16 infection by inducing noninfectious internalization. Mutation of a putative CyP binding site present in HPV16 L2 yielded exposed L2 N-terminus in the absence of active CyP and bypassed the need for cell surface CyPB. However, this mutant was still sensitive to CyP inhibition and required CyP for completion of infection, probably after internalization. Taken together, these data suggest that CyP is required during two distinct steps of HPV16 infection. Identification of cell surface CyPB will facilitate the study of the complex events preceding internalization and adds a putative drug target for prevention of HPV-induced diseases.

  8. Target cell cyclophilins facilitate human papillomavirus type 16 infection.

    Directory of Open Access Journals (Sweden)

    Malgorzata Bienkowska-Haba

    2009-07-01

    Full Text Available Following attachment to primary receptor heparan sulfate proteoglycans (HSPG, human papillomavirus type 16 (HPV16 particles undergo conformational changes affecting the major and minor capsid proteins, L1 and L2, respectively. This results in exposure of the L2 N-terminus, transfer to uptake receptors, and infectious internalization. Here, we report that target cell cyclophilins, peptidyl-prolyl cis/trans isomerases, are required for efficient HPV16 infection. Cell surface cyclophilin B (CyPB facilitates conformational changes in capsid proteins, resulting in exposure of the L2 N-terminus. Inhibition of CyPB blocked HPV16 infection by inducing noninfectious internalization. Mutation of a putative CyP binding site present in HPV16 L2 yielded exposed L2 N-terminus in the absence of active CyP and bypassed the need for cell surface CyPB. However, this mutant was still sensitive to CyP inhibition and required CyP for completion of infection, probably after internalization. Taken together, these data suggest that CyP is required during two distinct steps of HPV16 infection. Identification of cell surface CyPB will facilitate the study of the complex events preceding internalization and adds a putative drug target for prevention of HPV-induced diseases.

  9. Cyclophilin 40 facilitates HSP90-mediated RISC assembly in plants.

    Science.gov (United States)

    Iki, Taichiro; Yoshikawa, Manabu; Meshi, Tetsuo; Ishikawa, Masayuki

    2012-01-18

    Posttranscriptional gene silencing is mediated by RNA-induced silencing complexes (RISCs) that contain AGO proteins and single-stranded small RNAs. The assembly of plant AGO1-containing RISCs depends on the molecular chaperone HSP90. Here, we demonstrate that cyclophilin 40 (CYP40), protein phosphatase 5 (PP5), and several other proteins with the tetratricopeptide repeat (TPR) domain associates with AGO1 in an HSP90-dependent manner in extracts of evacuolated tobacco protoplasts (BYL). Intriguingly, CYP40, but not the other TPR proteins, could form a complex with small RNA duplex-bound AGO1. Moreover, CYP40 that was synthesized by in-vitro translation using BYL uniquely facilitated binding of small RNA duplexes to AGO1, and as a result, increased the amount of mature RISCs that could cleave target RNAs. CYP40 was not contained in mature RISCs, indicating that the association is transient. Addition of PP5 or cyclophilin-binding drug cyclosporine A prevented the association of endogenous CYP40 with HSP90-AGO1 complex and inhibited RISC assembly. These results suggest that a complex of AGO1, HSP90, CYP40, and a small RNA duplex is a key intermediate of RISC assembly in plants.

  10. Structural and Biochemical Characterization of the Human Cyclophilin Family of Peptidyl-Prolyl Isomerases

    Energy Technology Data Exchange (ETDEWEB)

    Davis, Tara L.; Walker, John R.; Campagna-Slater, Valérie; Finerty, Jr., Patrick J.; Paramanathan, Ragika; Bernstein, Galina; MacKenzie, Farrell; Tempel, Wolfram; Ouyang, Hui; Lee, Wen Hwa; Eisenmesser, Elan Z.; Dhe-Paganon, Sirano (Toronto); (Colorado)

    2011-12-14

    Peptidyl-prolyl isomerases catalyze the conversion between cis and trans isomers of proline. The cyclophilin family of peptidyl-prolyl isomerases is well known for being the target of the immunosuppressive drug cyclosporin, used to combat organ transplant rejection. There is great interest in both the substrate specificity of these enzymes and the design of isoform-selective ligands for them. However, the dearth of available data for individual family members inhibits attempts to design drug specificity; additionally, in order to define physiological functions for the cyclophilins, definitive isoform characterization is required. In the current study, enzymatic activity was assayed for 15 of the 17 human cyclophilin isomerase domains, and binding to the cyclosporin scaffold was tested. In order to rationalize the observed isoform diversity, the high-resolution crystallographic structures of seven cyclophilin domains were determined. These models, combined with seven previously solved cyclophilin isoforms, provide the basis for a family-wide structure:function analysis. Detailed structural analysis of the human cyclophilin isomerase explains why cyclophilin activity against short peptides is correlated with an ability to ligate cyclosporin and why certain isoforms are not competent for either activity. In addition, we find that regions of the isomerase domain outside the proline-binding surface impart isoform specificity for both in vivo substrates and drug design. We hypothesize that there is a well-defined molecular surface corresponding to the substrate-binding S2 position that is a site of diversity in the cyclophilin family. Computational simulations of substrate binding in this region support our observations. Our data indicate that unique isoform determinants exist that may be exploited for development of selective ligands and suggest that the currently available small-molecule and peptide-based ligands for this class of enzyme are insufficient for isoform

  11. Cyclophilin C Participates in the US2-Mediated Degradation of Major Histocompatibility Complex Class I Molecules.

    Science.gov (United States)

    Chapman, Daniel C; Stocki, Pawel; Williams, David B

    2015-01-01

    Human cytomegalovirus uses a variety of mechanisms to evade immune recognition through major histocompatibility complex class I molecules. One mechanism mediated by the immunoevasin protein US2 causes rapid disposal of newly synthesized class I molecules by the endoplasmic reticulum-associated degradation pathway. Although several components of this degradation pathway have been identified, there are still questions concerning how US2 targets class I molecules for degradation. In this study we identify cyclophilin C, a peptidyl prolyl isomerase of the endoplasmic reticulum, as a component of US2-mediated immune evasion. Cyclophilin C could be co-isolated with US2 and with the class I molecule HLA-A2. Furthermore, it was required at a particular expression level since depletion or overexpression of cyclophilin C impaired the degradation of class I molecules. To better characterize the involvement of cyclophilin C in class I degradation, we used LC-MS/MS to detect US2-interacting proteins that were influenced by cyclophilin C expression levels. We identified malectin, PDIA6, and TMEM33 as proteins that increased in association with US2 upon cyclophilin C knockdown. In subsequent validation all were shown to play a functional role in US2 degradation of class I molecules. This was specific to US2 rather than general ER-associated degradation since depletion of these proteins did not impede the degradation of a misfolded substrate, the null Hong Kong variant of α1-antitrypsin.

  12. Plasma level of cyclophilin A is increased in patients with type 2 diabetes mellitus and suggests presence of vascular disease.

    Science.gov (United States)

    Ramachandran, Surya; Venugopal, Anila; Kutty, V Raman; A, Vinitha; G, Divya; Chitrasree, V; Mullassari, Ajit; Pratapchandran, N S; Santosh, K R; Pillai, M Radhakrishna; Kartha, C C

    2014-02-07

    Cyclophilin A, an immunophilin is secreted from human monocytes activated by high glucose. Given its role as an inflammatory mediator of vascular tissue damage associated with inflammation and oxidative stress, we examined plasma levels of cyclophilin A in normal healthy volunteers and patients with type 2 diabetes (DM), with or without coronary artery disease (CAD). Study subjects comprised of 212 patients with DM and CAD,101 patients with diabetes, 122 patients with CAD and 121 normal healthy volunteers. Diabetes was assessed by HbA1c levels while coronary artery disease was established by a positive treadmill test and/or coronary angiography. Plasma cyclophilin A was measured using a cyclophilin A ELISA Kit. Relationship of plasma cyclophilin A levels with blood markers of type 2 diabetes, blood lipid levels and medication for diabetes and coronary artery disease were also explored. Plasma Cyclophilin levels were higher in diabetes patients with or without CAD compared to normal subjects (P levels and HbA1C levels were positively associated with increased plasma cyclophilin. Patients using metformin had reduced levels of plasma cyclophilin (p levels of total cholesterol, LDL cholesterol and triglycerides had no significant association with plasma cyclophilin levels. In patients with increased serum CRP levels, plasma cyclophilin A was also elevated (p = 0.016). Prevalence odds for DM, DM + CAD and CAD are higher in those with high cyclophilin values, compared to those with lower values, after adjusting for age and sex, indicating strong association of high cyclophilin values with diabetes and vascular disease. Our study demonstrates that patients with type 2 diabetes have higher circulating levels of cyclophilin A than the normal population. Plasma cyclophilin levels were increased in patients with diabetes and coronary artery disease suggesting a role of this protein in accelerating vascular disease in type 2 diabetes. Considering the evidence that

  13. Cyclophilin B expression in renal proximal tubules of hypertensive rats.

    Science.gov (United States)

    Kainer, D B; Doris, P A

    2000-04-01

    Rat cyclophilin-like protein (Cy-LP) is a candidate hypertension gene initially identified by differential hybridization and implicated in renal mechanisms of salt retention and high blood pressure. We report the molecular characterization of rat cyclophilin B (CypB) and demonstrate, through sequence analysis and an allele-specific polymerase chain reaction primer assay, that CypB but not Cy-LP is expressed in rat kidney. CypB is an endoplasmic reticulum-localized prolyl-isomerase that interacts with elongation initiation factor 2-beta, an important regulator of protein translation and a central component of the endoplasmic reticulum stress response to hypoxia or ATP depletion. Active renal transport of sodium is increased in the spontaneously hypertensive rat (SHR), and there is evidence that this coincides with hypoxia and ATP depletion in the renal cortex. In the present studies we have examined expression of CypB in rat proximal tubules, which contributes to the increased renal sodium reabsorption in this model of hypertension. We report that CypB transcript abundance is significantly elevated in proximal convoluted tubules from SHR compared with the control Wistar-Kyoto strain. This upregulation occurs in weanling animals and precedes the development of hypertension, indicating that it is not a simple response to hypertension in SHR. Further, CypB expression is also higher in a proximal tubule cell line derived from SHR compared with a similar line derived from Wistar-Kyoto rats, indicating that this difference is genetically determined. No sequence differences were observed in the CypB cDNA from these 2 strains. These observations suggest that a genetically determined alteration in proximal tubules from SHR occurs that leads to increased expression of CypB. In view of evidence linking CypB to the regulation of elongation initiation factor-2, the upregulation of CypB may result from metabolic stress.

  14. Computational insight into small molecule inhibition of cyclophilins.

    Science.gov (United States)

    Sambasivarao, Somisetti V; Acevedo, Orlando

    2011-02-28

    Cyclophilins (Cyp) are a family of cellular enzymes possessing peptidyl-prolyl isomerase activity, which catalyze the cis-trans interconversion of proline-containing peptide bonds. The two most abundant family members, CypA and CypB, have been identified as valid drug targets for a wide range of diseases, including HCV, HIV, and multiple cancers. However, the development of small molecule inhibitors that possess nM potency and high specificity for a particular Cyp is difficult given the complete conservation of all active site residues between the enzymes. Monte Carlo statistical sampling coupled to free energy perturbation theory (MC/FEP) calculations have been carried out to elucidate the origin of the experimentally observed nM inhibition of CypA by acylurea-based derivatives and the >200-fold in vitro selectivity between CypA and CypB from aryl 1-indanylketone-based μM inhibitors. The computed free-energies of binding were in close accord with those derived from experiments. Binding affinity values for the inhibitors were determined to be dependent upon the stabilization strength of the nonbonded interactions provided toward two catalytic residues: Arg55 and Asn102 in CypA and the analogous Arg63 and Asn110 residues in CypB. Fine-tuning of the hydrophobic interactions allowed for enhanced potency among derivatives. The aryl 1-indanylketones are predicted to differentiate between the cyclophilins by using distinct binding motifs that exploit subtle differences in the active site arrangements. Ideas for the development of new selective compounds with the potential for advancement to low-nanomolar inhibition are presented.

  15. Toxoplasma gondii infection in humans in China

    Directory of Open Access Journals (Sweden)

    He Shenyi

    2011-08-01

    Full Text Available Abstract Toxoplasmosis is a zoonotic infection of humans and animals, caused by the opportunistic protozoan Toxoplasma gondii, a parasite belonging to the phylum Apicomplexa. Infection in pregnant women may lead to abortion, stillbirth or other serious consequences in newborns. Infection in immunocompromised patients can be fatal if not treated. On average, one third of people are chronically infected worldwide. Although very limited information from China has been published in the English journals, T. gondii infection is actually a significant human health problem in China. In the present article, we reviewed the clinical features, transmission, prevalence of T. gondii infection in humans in China, and summarized genetic characterizations of reported T. gondii isolates. Educating the public about the risks associated with unhealthy food and life style habits, tracking serological examinations to special populations, and measures to strengthen food and occupational safety are discussed.

  16. Molecular detection of Toxoplasma gondii in snakes.

    Science.gov (United States)

    Nasiri, Vahid; Teymurzadeh, Shohreh; Karimi, Gholamreza; Nasiri, Mehdi

    2016-10-01

    Toxoplasma gondii, an obligate intracellular protozoan parasite, is responsible for one of the most common zoonotic parasitic diseases in almost all warm-blooded vertebrates worldwide, and it is estimated that about one-third of the world human population is chronically infected with this parasite. Little is known about the circulation of T. gondii in snakes and this study for the first time aimed to evaluate the infection rates of snakes by this parasite by PCR methods. The brain of 68 Snakes, that were collected between May 2012 and September 2015 and died after the hold in captivity, under which they were kept for taking poisons, were examined for the presence of this parasite. DNA was extracted and Nested-PCR method was carried out with two of pairs of primers to detect the 344 bp fragment of T. gondii GRA6 gene. Five positive nested-PCR products were directly sequenced in the forward and reverse directions by Sequetech Company (Mountain View, CA). T. gondii GRA6 gene were detected from 55 (80.88%) of 68 snakes brains. Sequencing of the GRA6 gene revealed 98-100% of similarity with T. gondii sequences deposited in GenBank. To our knowledge, this is the first study of molecular detection of T. gondii in snakes and our findings show a higher frequency of this organism among them. Copyright © 2016 Elsevier Inc. All rights reserved.

  17. Cloning and expression of Toxoplasma gondii tachyzoite P22 protein

    African Journals Online (AJOL)

    Jane

    2011-08-01

    Aug 1, 2011 ... Expressd protein was purified by affinity chromatography and confirmed by western blot ... Key words: Toxoplasma gondii, cloning, recombinant P22. INTRODUCTION. Toxoplasma gondii ..... an ELISA Assay. Iran. J. Immunol.

  18. Toxoplasma gondii seroprevalence in breeding pigs in Estonia

    DEFF Research Database (Denmark)

    Santoro, Azzurra; Tagel, Maarja; Must, Kärt

    2017-01-01

    Background: Toxoplasma gondii is a widespread occurring parasite infecting warm-blooded animals, including pigs and humans. The aims of this study were to estimate the prevalence of anti-T. gondii antibodies and to evaluate risk factors for T. gondii seropositivity in breeding pigs raised in Esto...

  19. Genetic diversity of Toxoplama gondii isolates from Ethiopian feral cats

    Science.gov (United States)

    Recent studies indicate greater genetic variability among isolates of Toxoplasma gondii worldwide than previously thought. However, there is no information on genetic diversity of T. gondii from any host in Ethiopia. In the present study, genotyping was performed on viable T. gondii isolates by bioa...

  20. Cyclophilin B Deficiency Causes Abnormal Dentin Collagen Matrix.

    Science.gov (United States)

    Terajima, Masahiko; Taga, Yuki; Cabral, Wayne A; Nagasawa, Masako; Sumida, Noriko; Hattori, Shunji; Marini, Joan C; Yamauchi, Mitsuo

    2017-08-04

    Cyclophilin B (CypB) is an endoplasmic reticulum-resident protein that regulates collagen folding, and also contributes to prolyl 3-hydroxylation (P3H) and lysine (Lys) hydroxylation of collagen. In this study, we characterized dentin type I collagen in CypB null (KO) mice, a model of recessive osteogenesis imperfecta type IX, and compared to those of wild-type (WT) and heterozygous (Het) mice. Mass spectrometric analysis demonstrated that the extent of P3H in KO collagen was significantly diminished compared to WT/Het. Lys hydroxylation in KO was significantly diminished at the helical cross-linking sites, α1/α2(I) Lys-87 and α1(I) Lys-930, leading to a significant increase in the under-hydroxylated cross-links and a decrease in fully hydroxylated cross-links. The extent of glycosylation of hydroxylysine residues was, except α1(I) Lys-87, generally higher in KO than WT/Het. Some of these molecular phenotypes were distinct from other KO tissues reported previously, indicating the dentin-specific control mechanism through CypB. Histological analysis revealed that the width of predentin was greater and irregular, and collagen fibrils were sparse and significantly smaller in KO than WT/Het. These results indicate a critical role of CypB in dentin matrix formation, suggesting a possible association between recessive osteogenesis imperfecta and dentin defects that have not been clinically detected.

  1. The Roles of CD147 and/or Cyclophilin A in Kidney Diseases

    Science.gov (United States)

    Wang, Chunting; Zhang, Jicheng; Qie, Guoqiang

    2014-01-01

    CD147 is a widely expressed integral plasma membrane glycoprotein and has been involved in a variety of physiological and pathological activities in combination with different partners, including cyclophilins, caveolin-1, monocarboxylate transporters, and integrins. Recent data demonstrate that both CyPA and CD147 significantly contribute to renal inflammation, acute kidney injury, renal fibrosis, and renal cell carcinoma. Here we review the current understanding of cyclophilin A and CD147 expression and functions in kidney diseases and potential implications for treatment of kidney diseases. PMID:25580061

  2. The Roles of CD147 and/or Cyclophilin A in Kidney Diseases

    Directory of Open Access Journals (Sweden)

    Xin Qu

    2014-01-01

    Full Text Available CD147 is a widely expressed integral plasma membrane glycoprotein and has been involved in a variety of physiological and pathological activities in combination with different partners, including cyclophilins, caveolin-1, monocarboxylate transporters, and integrins. Recent data demonstrate that both CyPA and CD147 significantly contribute to renal inflammation, acute kidney injury, renal fibrosis, and renal cell carcinoma. Here we review the current understanding of cyclophilin A and CD147 expression and functions in kidney diseases and potential implications for treatment of kidney diseases.

  3. Cyclophilin B stimulates RNA synthesis by the HCV RNA dependent RNA polymerase

    OpenAIRE

    Heck, Julie A.; Meng, Xiao; Frick, David N.

    2009-01-01

    Cyclophilins are cellular peptidyl isomerases that have been implicated in regulating hepatitis C virus (HCV) replication. Cyclophilin B (CypB) is a target of cyclosporin A (CsA), an immunosuppressive drug recently shown to suppress HCV replication in cell culture. Watashi et al. recently demonstrated that CypB is important for efficient HCV replication, and proposed that it mediates the anti-HCV effects of CsA through an interaction with NS5B (Mol. Cell 19:111). We examined the effects of pu...

  4. The transcriptome of Toxoplasma gondii

    Directory of Open Access Journals (Sweden)

    Roos David S

    2005-12-01

    Full Text Available Abstract Background Toxoplasma gondii gives rise to toxoplasmosis, among the most prevalent parasitic diseases of animals and man. Transformation of the tachzyoite stage into the latent bradyzoite-cyst form underlies chronic disease and leads to a lifetime risk of recrudescence in individuals whose immune system becomes compromised. Given the importance of tissue cyst formation, there has been intensive focus on the development of methods to study bradyzoite differentiation, although the molecular basis for the developmental switch is still largely unknown. Results We have used serial analysis of gene expression (SAGE to define the Toxoplasma gondii transcriptome of the intermediate-host life cycle that leads to the formation of the bradyzoite/tissue cyst. A broad view of gene expression is provided by >4-fold coverage from nine distinct libraries (~300,000 SAGE tags representing key developmental transitions in primary parasite populations and in laboratory strains representing the three canonical genotypes. SAGE tags, and their corresponding mRNAs, were analyzed with respect to abundance, uniqueness, and antisense/sense polarity and chromosome distribution and developmental specificity. Conclusion This study demonstrates that phenotypic transitions during parasite development were marked by unique stage-specific mRNAs that accounted for 18% of the total SAGE tags and varied from 1–5% of the tags in each developmental stage. We have also found that Toxoplasma mRNA pools have a unique parasite-specific composition with 1 in 5 transcripts encoding Apicomplexa-specific genes functioning in parasite invasion and transmission. Developmentally co-regulated genes were dispersed across all Toxoplasma chromosomes, as were tags representing each abundance class, and a variety of biochemical pathways indicating that trans-acting mechanisms likely control gene expression in this parasite. We observed distinct similarities in the specificity and

  5. Crioconservación de toxoplasma gondii

    Directory of Open Access Journals (Sweden)

    Sofía Duque Beltrán

    1992-03-01

    Full Text Available Toxoplasma gondii fue crioconservado en nitrógeno líquido usando como preservativo glicerol al 10% con el fin de mantener el protozoo por un largo período de tiempo. El descongelamiento de T. gondii se llevó a cabo, cuando los parásitos fueron requeridos para uso como antígeno, a los 10, 40 y 270 días siguientes a su crioconservación. La viabilidad y patogenicidad del parásito fue confirmada in vivo. La crioconservación de T. gondii disminuyó los costos de mantenimiento in vivo y de recursos humanos tanto en el bioterio como en el laboratorio.

  6. Sexual transmission of Toxoplasma gondii in sheep.

    Science.gov (United States)

    Lopes, Welber Daniel Zanetti; Rodriguez, Joana D'Ark; Souza, Fernando A; dos Santos, Thais Rabelo; dos Santos, Ricardo Silva; Rosanese, Walter Matheus; Lopes, Werik Renato Zanetti; Sakamoto, Cláudio Alessandro; da Costa, Alvimar José

    2013-07-01

    Male sheep of reproductive age were distributed into three groups: GI, a sheep inoculated (oral) with 2.0×10(5) oocysts of the P strain of Toxoplasma gondii; GII, a sheep infected (subcutaneous) with 1.0×10(6) tachyzoites of the RH strain of T. gondii; and GIII, a sheep kept as a control (not infected). After the inoculation of the males, 12 breeding ewes, which were not pregnant and which were serologically negative for reproductive diseases (particularly toxoplasmosis), were distributed into three groups, synchronized, and subsequently exposed to natural mating with previously inoculated males. The distribution was as follows: five ewes that underwent natural mating with the GI male, five ewes that were exposed to natural mating with the GII male, and two ewes that were mated with the non-infected male (control). Serum samples of all the ewes were collected on days -30, -14, -7, -1, and 0 (days before natural mating) and on days 1, 3, 5, 7, 11, 14, and weekly until birth; the presence of serum antibodies against T. gondii was assessed by IFAT. Using a bioassay and PCR, T. gondii was isolated from the semen of the infected reproducing sheep before mating. Following natural mating, 5 of the 12 females displayed antibodies specific for T. gondii; of these animals, two of the ewes underwent natural mating with the male inoculated with oocysts (GI) and three with the male infected with tachyzoites (GII). One of the females that displayed antibodies specific to this coccidian and that underwent natural mating with the GII sheep had a macerated fetus on the 70th day following coverage. Using a bioassay after the birth, it was possible to isolate T. gondii from samples of the "pool" of tissues from the five females that seroconverted after natural mating and from their respective lambs. Using PCR, the DNA of T. gondii was isolated from the "pool" of tissues from one and two females exposed to natural mating with the reproductive males infected with the oocysts and

  7. Crystal structures of wild-type and mutated cyclophilin B that causes hyperelastosis cutis in the American quarter horse

    Directory of Open Access Journals (Sweden)

    Boudko Sergei P

    2012-11-01

    Full Text Available Abstract Background Hyperelastosis cutis is an inherited autosomal recessive connective tissue disorder. Affected horses are characterized by hyperextensible skin, scarring, and severe lesions along the back. The disorder is caused by a mutation in cyclophilin B. Results The crystal structures of both wild-type and mutated (Gly6->Arg horse cyclophilin B are presented. The mutation neither affects the overall fold of the enzyme nor impairs the catalytic site structure. Instead, it locally rearranges the flexible N-terminal end of the polypeptide chain and also makes it more rigid. Conclusions Interactions of the mutated cyclophilin B with a set of endoplasmic reticulum-resident proteins must be affected.

  8. Delineation of the calcineurin-interacting region of cyclophilin B.

    Science.gov (United States)

    Carpentier, M; Allain, F; Haendler, B; Slomianny, M C; Spik, G

    2000-12-01

    The immunosuppressant drug cyclosporin A (CsA) inhibits T-cell function by blocking the phosphatase activity of calcineurin. This effect is mediated by formation of a complex between the drug and cyclophilin (CyP), which creates a composite surface able to make high-affinity contacts with calcineurin. In vitro, the CyPB/CsA complex is more effective in inhibiting calcineurin than the CyPA/CsA and CyPC/CsA complexes, pointing to fine structural differences in the calcineurin-binding region. To delineate the calcineurin-binding region of CyPB, we mutated several amino acids, located in two loops corresponding to CyPA regions known to be involved, as follows: R76A, G77H, D155R, and D158R. Compared to wild-type CyPB, the G77H, D155R, and D158R mutants had intact isomerase and CsA-binding activities, indicating that no major conformational changes had taken place. When complexed to CsA, they all displayed only reduced affinity for calcineurin and much decreased inhibition of calcineurin phosphatase activity. These results strongly suggest that the three amino acids G77, D155, and D158 are directly involved in the interaction of CyPB/CsA with calcineurin, in agreement with their exposed position. The G77, D155, and D158 residues are not maintained in CyPA and might therefore account for the higher affinity of the CyPB/CsA complex for calcineurin.

  9. Cyclophilin B facilitates the replication of Orf virus.

    Science.gov (United States)

    Zhao, Kui; Li, Jida; He, Wenqi; Song, Deguang; Zhang, Ximu; Zhang, Di; Zhou, Yanlong; Gao, Feng

    2017-06-15

    Viruses interact with host cellular factors to construct a more favourable environment for their efficient replication. Expression of cyclophilin B (CypB), a cellular peptidyl-prolyl cis-trans isomerase (PPIase), was found to be significantly up-regulated. Recently, a number of studies have shown that CypB is important in the replication of several viruses, including Japanese encephalitis virus (JEV), hepatitis C virus (HCV) and human papillomavirus type 16 (HPV 16). However, the function of cellular CypB in ORFV replication has not yet been explored. Suppression subtractive hybridization (SSH) technique was applied to identify genes differentially expressed in the ORFV-infected MDBK cells at an early phase of infection. Cellular CypB was confirmed to be significantly up-regulated by quantitative reverse transcription-PCR (qRT-PCR) analysis and Western blotting. The role of CypB in ORFV infection was further determined using Cyclosporin A (CsA) and RNA interference (RNAi). Effect of CypB gene silencing on ORFV replication by 50% tissue culture infectious dose (TCID 50 ) assay and qRT-PCR detection. In the present study, CypB was found to be significantly up-regulated in the ORFV-infected MDBK cells at an early phase of infection. Cyclosporin A (CsA) exhibited suppressive effects on ORFV replication through the inhibition of CypB. Silencing of CypB gene inhibited the replication of ORFV in MDBK cells. In conclusion, these data suggest that CypB is critical for the efficient replication of the ORFV genome. Cellular CypB was confirmed to be significantly up-regulated in the ORFV-infected MDBK cells at an early phase of infection, which could effectively facilitate the replication of ORFV.

  10. Mitochondrial Cyclophilin D in Vascular Oxidative Stress and Hypertension.

    Science.gov (United States)

    Itani, Hana A; Dikalova, Anna E; McMaster, William G; Nazarewicz, Rafal R; Bikineyeva, Alfiya T; Harrison, David G; Dikalov, Sergey I

    2016-06-01

    Vascular superoxide (O˙2 (-)) and inflammation contribute to hypertension. The mitochondria are an important source of O˙2 (-); however, the regulation of mitochondrial O˙2 (-) and the antihypertensive potential of targeting the mitochondria remain poorly defined. Angiotensin II and inflammatory cytokines, such as interleukin 17A and tumor necrosis factor-α (TNFα) significantly contribute to hypertension. We hypothesized that angiotensin II and cytokines co-operatively induce cyclophilin D (CypD)-dependent mitochondrial O˙2 (-) production in hypertension. We tested whether CypD inhibition attenuates endothelial oxidative stress and reduces hypertension. CypD depletion in CypD(-/-) mice prevents overproduction of mitochondrial O˙2 (-) in angiotensin II-infused mice, attenuates hypertension by 20 mm Hg, and improves vascular relaxation compared with wild-type C57Bl/6J mice. Treatment of hypertensive mice with the specific CypD inhibitor Sanglifehrin A reduces blood pressure by 28 mm Hg, inhibits production of mitochondrial O˙2 (-) by 40%, and improves vascular relaxation. Angiotensin II-induced hypertension was associated with CypD redox activation by S-glutathionylation, and expression of the mitochondria-targeted H2O2 scavenger, catalase, abolished CypD S-glutathionylation, prevented stimulation mitochondrial O˙2 (-), and attenuated hypertension. The functional role of cytokine-angiotensin II interplay was confirmed by co-operative stimulation of mitochondrial O˙2 (-) by 3-fold in cultured endothelial cells and impairment of aortic relaxation incubated with combination of angiotensin II, interleukin 17A, and tumor necrosis factor-α which was prevented by CypD depletion or expression of mitochondria-targeted SOD2 and catalase. These data support a novel role of CypD in hypertension and demonstrate that targeting CypD decreases mitochondrial O˙2 (-), improves vascular relaxation, and reduces hypertension. © 2016 American Heart Association, Inc.

  11. Cloning and Expression of Cyclophilin from Platanus orientalis Pollens in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Mojtaba Sankian

    2012-10-01

    Full Text Available Background: Allergy is a clinical disorder affecting the human population with wide geographical distribution. Platanus orientalis (P. orientalis trees are planted in many countries and their pollen causes allergic reactions. Cyclophilin has recently been identified as one of the most important allergens of P. orientalis pollen. We aimed to clone and purify this allergen in Escherichia coli for further studies and therapeutic and diagnostic purposes for allergy to P. orientalis. Methods: RNA was extracted from P. orientalis. A full-length fragment encoding cyclophilin was prepared by polymerase chain reaction amplification of the first-strand cDNA synthesized from P. orientalis RNA. The cDNA was inserted into the pET32b (+ vector, and the construct transformed into E. coli Top10 and BL21 cells. The expressed protein was purified by the CuSO4 method. Results: The cDNA for the cyclophilin of P. orientalis pollen was cloned, and a specific reactivity of recombinant cyclophin was confirmed by immunoblotting using sera from patients allergic to P. orientalis pollen. Conclusion: The recombinant cyclophilin has a potential for immunologic assays for evaluation of allergy to P. orientalis pollen.

  12. Intracellularly Induced Cyclophilins Play an Important Role in Stress Adaptation and Virulence of Brucella abortus

    Science.gov (United States)

    García Fernández, Lucía; DelVecchio, Vito G.; Briones, Gabriel

    2013-01-01

    Brucella is an intracellular bacterial pathogen that causes the worldwide zoonotic disease brucellosis. Brucella virulence relies on its ability to transition to an intracellular lifestyle within host cells. Thus, this pathogen must sense its intracellular localization and then reprogram gene expression for survival within the host cell. A comparative proteomic investigation was performed to identify differentially expressed proteins potentially relevant for Brucella intracellular adaptation. Two proteins identified as cyclophilins (CypA and CypB) were overexpressed in the intracellular environment of the host cell in comparison to laboratory-grown Brucella. To define the potential role of cyclophilins in Brucella virulence, a double-deletion mutant was constructed and its resulting phenotype was characterized. The Brucella abortus ΔcypAB mutant displayed increased sensitivity to environmental stressors, such as oxidative stress, pH, and detergents. In addition, the B. abortus ΔcypAB mutant strain had a reduced growth rate at lower temperature, a phenotype associated with defective expression of cyclophilins in other microorganisms. The B. abortus ΔcypAB mutant also displays reduced virulence in BALB/c mice and defective intracellular survival in HeLa cells. These findings suggest that cyclophilins are important for Brucella virulence and survival in the host cells. PMID:23230297

  13. Seroprevalence of Toxoplasma gondii and Neospora caninum ...

    African Journals Online (AJOL)

    The aim of this study was to compare the seroprevalence of Toxoplasma gondii and Neospora caninum in goats from two Argentinean provinces raised under different management conditions. A total of 2922 serum samples from adult goats of Córdoba (n=2187) and Buenos Aires provinces (n= 735), Argentina, were ...

  14. Mechanics of the Toxoplasma gondii oocyst wall

    Science.gov (United States)

    The ability of microorganisms to survive under extreme conditions is closely related to the physicochemical properties of their wall. In the ubiquitous protozoan parasite Toxoplasma gondii, the oocyst stage possesses a bilayered wall that protects the dormant but potentially infective parasites from...

  15. Toxoplasma gondii gamma irradiation using Co-60

    International Nuclear Information System (INIS)

    Serra-Freire, Nicolau Maues

    1996-01-01

    The use of nuclear power through radiation for the destruction of microorganisms which cause food deterioration, infections and toxicosis, is specifically for peaceful purposes. Toxoplasma gondii is a protozoa responsible for illnesses in humans and animals. One of the most common ways of transmission is through raw or poorly cooked meat. There is little information on the resistance of T. gondii to radiation. The objective of this research is to determine the Minimum Lethal Dose (MLD) of gamma radiation for those microorganisms. Suspensions of T. gondii containing approximately one million taquizoites/ml were irradiated with doses between up 0,01 up to 0,15 kGy (Kilogray) and inoculated to mice. The surviving T. gondii were re-irradiated with 0,01 up to 0,16 kGy. The irradiated protozoa were totally destroyed with a 0,15 kGy dose (MLD). Taquizoites issued from live protozoa of 0,14 kGy also were completely destroyed with dose of 0,15 kGy. No increase in resistance was observed regarding the non irradiated protozoa. (author)

  16. Uncaria rhynchophylla upregulates the expression of MIF and cyclophilin A in kainic acid-induced epilepsy rats: A proteomic analysis.

    Science.gov (United States)

    Lo, Wan-Yu; Tsai, Fuu-Jen; Liu, Chung-Hsiang; Tang, Nou-Ying; Su, Shan-Yu; Lin, Shinn-Zong; Chen, Chun-Chung; Shyu, Woei-Cherng; Hsieh, Ching-Liang

    2010-01-01

    Uncaria rhynchophylla (Miq) Jack (UR) is a traditional Chinese herb and is used for the treatment of convulsive disorders, including epilepsy. Our previous study has shown that UR, as well as its major component rhynchophylline (RH), has an anticonvulsive effect and this effect is closely related to its scavenging activities of oxygen free radicals. The purpose of the present study was to investigate the effect of (UR) on the expression of proteins using a proteomics analysis in Sprague-Dawley (SD) rats with kainic acid (KA)-induced epileptic seizures. We profiled the differentially expressed proteins on two-dimensional electrophoresis (2-DE) maps derived from the frontal cortex and hippocampus of rat brain tissue 24 hours after KA-induced epileptic seizures. The results indicated that macrophage migration inhibitory factor (MIF) and cyclophilin A were under expressed in frontal cortex by an average of 0.19- and 0.23-fold, respectively. In the frontal cortex, MIF and cyclophilin A were significantly decreased in the KA group and these decreases were confirmed by the Western blots. However, in the hippocampus, only cyclophilin A was significantly decreased in the KA group. In addition, in real-time quantitative PCR (Q-PCR), MIF and cyclophilin A gene expressions were also significantly under expressed in the frontal cortex, and only the cyclophilin A gene was also significantly under expressed in the hippocampus in the KA group. These under expressions of MIF and cyclophilin A could be overcome by the treatment of UR and RH. In conclusion, the under expressions of MIF and cyclophilin A in the frontal cortex and hippocampus in KA-treated rats, which were overcome by both UR and UH treatment, suggesting that both MIF and cyclophilin A at least partly participate in the anticonvulsive effect of UR.

  17. Severe osteogenesis imperfecta in cyclophilin B-deficient mice.

    Directory of Open Access Journals (Sweden)

    Jae Won Choi

    2009-12-01

    Full Text Available Osteogenesis Imperfecta (OI is a human syndrome characterized by exquisitely fragile bones due to osteoporosis. The majority of autosomal dominant OI cases result from point or splice site mutations in the type I collagen genes, which are thought to lead to aberrant osteoid within developing bones. OI also occurs in humans with homozygous mutations in Prolyl-3-Hydroxylase-1 (LEPRE1. Although P3H1 is known to hydroxylate a single residue (pro-986 in type I collagen chains, it is unclear how this modification acts to facilitate collagen fibril formation. P3H1 exists in a complex with CRTAP and the peptidyl-prolyl isomerase cyclophilin B (CypB, encoded by the Ppib gene. Mutations in CRTAP cause OI in mice and humans, through an unknown mechanism, while the role of CypB in this complex has been a complete mystery. To study the role of mammalian CypB, we generated mice lacking this protein. Early in life, Ppib-/- mice developed kyphosis and severe osteoporosis. Collagen fibrils in Ppib-/- mice had abnormal morphology, further consistent with an OI phenotype. In vitro studies revealed that in CypB-deficient fibroblasts, procollagen did not localize properly to the golgi. We found that levels of P3H1 were substantially reduced in Ppib-/- cells, while CRTAP was unaffected by loss of CypB. Conversely, knockdown of either P3H1 or CRTAP did not affect cellular levels of CypB, but prevented its interaction with collagen in vitro. Furthermore, knockdown of CRTAP also caused depletion of cellular P3H1. Consistent with these changes, post translational prolyl-3-hydroxylation of type I collagen by P3H1 was essentially absent in CypB-deficient cells and tissues from CypB-knockout mice. These data provide significant new mechanistic insight into the pathophysiology of OI and reveal how the members of the P3H1/CRTAP/CypB complex interact to direct proper formation of collagen and bone.

  18. Severe Osteogenesis Imperfecta in Cyclophilin B–Deficient Mice

    Science.gov (United States)

    Choi, Jae Won; Sutor, Shari L.; Lindquist, Lonn; Evans, Glenda L.; Madden, Benjamin J.; Bergen, H. Robert; Hefferan, Theresa E.; Yaszemski, Michael J.; Bram, Richard J.

    2009-01-01

    Osteogenesis Imperfecta (OI) is a human syndrome characterized by exquisitely fragile bones due to osteoporosis. The majority of autosomal dominant OI cases result from point or splice site mutations in the type I collagen genes, which are thought to lead to aberrant osteoid within developing bones. OI also occurs in humans with homozygous mutations in Prolyl-3-Hydroxylase-1 (LEPRE1). Although P3H1 is known to hydroxylate a single residue (pro-986) in type I collagen chains, it is unclear how this modification acts to facilitate collagen fibril formation. P3H1 exists in a complex with CRTAP and the peptidyl-prolyl isomerase cyclophilin B (CypB), encoded by the Ppib gene. Mutations in CRTAP cause OI in mice and humans, through an unknown mechanism, while the role of CypB in this complex has been a complete mystery. To study the role of mammalian CypB, we generated mice lacking this protein. Early in life, Ppib-/- mice developed kyphosis and severe osteoporosis. Collagen fibrils in Ppib-/- mice had abnormal morphology, further consistent with an OI phenotype. In vitro studies revealed that in CypB–deficient fibroblasts, procollagen did not localize properly to the golgi. We found that levels of P3H1 were substantially reduced in Ppib-/- cells, while CRTAP was unaffected by loss of CypB. Conversely, knockdown of either P3H1 or CRTAP did not affect cellular levels of CypB, but prevented its interaction with collagen in vitro. Furthermore, knockdown of CRTAP also caused depletion of cellular P3H1. Consistent with these changes, post translational prolyl-3-hydroxylation of type I collagen by P3H1 was essentially absent in CypB–deficient cells and tissues from CypB–knockout mice. These data provide significant new mechanistic insight into the pathophysiology of OI and reveal how the members of the P3H1/CRTAP/CypB complex interact to direct proper formation of collagen and bone. PMID:19997487

  19. Severe osteogenesis imperfecta in cyclophilin B-deficient mice.

    Science.gov (United States)

    Choi, Jae Won; Sutor, Shari L; Lindquist, Lonn; Evans, Glenda L; Madden, Benjamin J; Bergen, H Robert; Hefferan, Theresa E; Yaszemski, Michael J; Bram, Richard J

    2009-12-01

    Osteogenesis Imperfecta (OI) is a human syndrome characterized by exquisitely fragile bones due to osteoporosis. The majority of autosomal dominant OI cases result from point or splice site mutations in the type I collagen genes, which are thought to lead to aberrant osteoid within developing bones. OI also occurs in humans with homozygous mutations in Prolyl-3-Hydroxylase-1 (LEPRE1). Although P3H1 is known to hydroxylate a single residue (pro-986) in type I collagen chains, it is unclear how this modification acts to facilitate collagen fibril formation. P3H1 exists in a complex with CRTAP and the peptidyl-prolyl isomerase cyclophilin B (CypB), encoded by the Ppib gene. Mutations in CRTAP cause OI in mice and humans, through an unknown mechanism, while the role of CypB in this complex has been a complete mystery. To study the role of mammalian CypB, we generated mice lacking this protein. Early in life, Ppib-/- mice developed kyphosis and severe osteoporosis. Collagen fibrils in Ppib-/- mice had abnormal morphology, further consistent with an OI phenotype. In vitro studies revealed that in CypB-deficient fibroblasts, procollagen did not localize properly to the golgi. We found that levels of P3H1 were substantially reduced in Ppib-/- cells, while CRTAP was unaffected by loss of CypB. Conversely, knockdown of either P3H1 or CRTAP did not affect cellular levels of CypB, but prevented its interaction with collagen in vitro. Furthermore, knockdown of CRTAP also caused depletion of cellular P3H1. Consistent with these changes, post translational prolyl-3-hydroxylation of type I collagen by P3H1 was essentially absent in CypB-deficient cells and tissues from CypB-knockout mice. These data provide significant new mechanistic insight into the pathophysiology of OI and reveal how the members of the P3H1/CRTAP/CypB complex interact to direct proper formation of collagen and bone.

  20. Long-term inhibition of cyclophilin D results in intracellular translocation of calcein AM from mitochondria to lysosomes.

    Science.gov (United States)

    Shinohe, Daisuke; Kobayashi, Asuka; Gotoh, Marina; Tanaka, Kotaro; Ohta, Yoshihiro

    2017-01-01

    Cyclophilin D is a peptidyl-prolyl cis-trans isomerase localized in the mitochondrial matrix. Although its effects on mitochondrial characteristics have been well studied, its relation to the uptake of molecules by mitochondria remains unknown. Here, we demonstrated the effects of cyclophilin D on the intracellular translocation of calcein AM. Following addition of calcein AM to control cells or cells overexpressing wild-type cyclophilin D, calcein fluorescence was observed in mitochondria. However, long-term inhibition of cyclophilin D in these cells altered the localization of calcein fluorescence from mitochondria to lysosomes without changing mitochondrial esterase activity. In addition, depletion of glucose from the medium recovered calcein localization from lysosomes to mitochondria. This is the first demonstration of the effects of cyclophilin D on the intracellular translocation of molecules other than proteins and suggests that cyclophilin D may modify mitochondrial features by inducing the translocation of molecules to the mitochondria through the mechanism associated with cellular energy metabolism. Copyright © 2016 Elsevier Inc. All rights reserved.

  1. 21 CFR 866.3780 - Toxoplasma gondii serological reagents.

    Science.gov (United States)

    2010-04-01

    ... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3780 Toxoplasma... (immunofluorescent reagents) used to identify Toxoplasma gondii from clinical specimens. The identification aids in...

  2. Toxoplasma gondii in small neotropical wild felids

    Directory of Open Access Journals (Sweden)

    William Alberto Cañon-Franco

    2013-02-01

    Full Text Available In the last decade, studies on wildlife worldwide have discovered key epidemiological aspects of the sylvatic cycle of Toxoplasma gondii. However, despite the known role of wild felines as definitive hosts in the transmission and maintenance of this parasite, few studies have focused on the involvement of these animals. Brazil exhibits the largest number of wild felid species in the Americas, all of which have a critical conservation status. However, serological detections, epidemiological studies and some molecular characterizations of T. gondii have primarily used Neotropical felid populations that are maintained in captivity, which does not reflect the disease behavior in free-living conditions. A systematic review of the worldwide scientific literature was conducted focusing on toxoplasmosis in small Neotropical felids. This review covered a number of aspects, including the state of scientific research, parasite transmission in the wild, the genetic characteristics of isolates, the relationship between these genetic characteristics and the pathogenicity of the parasite, and the risk factors linked to conflicts with humans. The present review shows the relevance of studying these felid populations based on their frequent interactions with humans in peri-urban areas and the need for further comprehensive studies to establish the real significance of T. gondii in public and animal health in tropical and temperate regions.

  3. A proteome study of secreted prostatic factors affecting osteoblastic activity, identification and characterisation of cyclophilin A

    DEFF Research Database (Denmark)

    Andersen, H.; Jensen, O.N.; Eriksen, E.F.

    2003-01-01

    ranging from 5 to 30 kDa were analysed by matrix-assisted laser desorption/ionisation time of flight mass spectrometry (MALDI-TOF MS). One of these spots was identified as cyclophilin A. We examined whether cyclophilin A alone or in combination with insulin-like growth factor-I (IGF-I) had any effects...... was to characterise the protein profile of conditioned medium (CM) from PC3 cells in the molecular weight range of 5-30 kDa using proteome analysis. A protein profile of the CM from PC3 cells was performed by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Thirty protein spots with molecular weights...

  4. Hypoxia induces cyclophilin B through the activation of transcription factor 6 in gastric adenocarcinoma cells

    OpenAIRE

    JEONG, KWON; KIM, KIYOON; KIM, HUNSUNG; OH, YOOJUNG; KIM, SEONG-JIN; JO, YUNHEE; CHOE, WONCHAE

    2015-01-01

    Hypoxia is an important form of physiological stress that induces cell death, due to the resulting endoplasmic reticulum (ER) stress, particularly in solid tumors. Although previous studies have indicated that cyclophilin B (CypB) plays a role in ER stress, there is currently no direct information supporting the mechanism of CypB involvement under hypoxic conditions. However, it has previously been demonstrated that ER stress positively regulates the expression of CypB. In the present study, ...

  5. Cyclophilin40 isomerase activity is regulated by a temperature-dependent allosteric interaction with Hsp90.

    Science.gov (United States)

    Blackburn, Elizabeth A; Wear, Martin A; Landré, Vivian; Narayan, Vikram; Ning, Jia; Erman, Burak; Ball, Kathryn L; Walkinshaw, Malcolm D

    2015-09-01

    Cyclophilin 40 (Cyp40) comprises an N-terminal cyclophilin domain with peptidyl-prolyl isomerase (PPIase) activity and a C-terminal tetratricopeptide repeat (TPR) domain that binds to the C-terminal-EEVD sequence common to both heat shock protein 70 (Hsp70) and Hsp90. We show in the present study that binding of peptides containing the MEEVD motif reduces the PPIase activity by ∼30%. CD and fluorescence assays show that the TPR domain is less stable than the cyclophilin domain and is stabilized by peptide binding. Isothermal titration calorimetry (ITC) shows that the affinity for the-MEEVD peptide is temperature sensitive in the physiological temperature range. Results from these biophysical studies fit with the MD simulations of the apo and holo (peptide-bound) structures which show a significant reduction in root mean square (RMS) fluctuation in both TPR and cyclophilin domains when-MEEVD is bound. The MD simulations of the apo-protein also highlight strong anti-correlated motions between residues around the PPIase-active site and a band of residues running across four of the seven helices in the TPR domain. Peptide binding leads to a distortion in the shape of the active site and a significant reduction in these strongly anti-correlated motions, providing an explanation for the allosteric effect of ligand binding and loss of PPIase activity. Together the experimental and MD results suggest that on heat shock, dissociation of Cyp40 from complexes mediated by the TPR domain leads to an increased pool of free Cyp40 capable of acting as an isomerase/chaperone in conditions of cellular stress. © 2015 Authors.

  6. Seroprevalence of Toxoplasma gondii infection in Norwegian dairy goats

    Directory of Open Access Journals (Sweden)

    Stormoen Marit

    2012-12-01

    Full Text Available Abstract Background Toxoplasma gondii is a major problem for the sheep industry as it may cause reproduction problems. The importance of T. gondii in Norwegian goat herds is uncertain, but outbreaks of toxoplasmosis in dairy goat farms have been recorded. The aim of this study was to describe the prevalence of T. gondii infection in Norwegian dairy goats by using serology. Findings Goat serum originally collected as part of two nationwide surveillance and control programmes between 2002 and 2008 were examined for T. gondii antibodies by using direct agglutination test. In total, 55 of 73 herds (75% had one or more serologically positive animals, while 377 of 2188 (17% of the individual samples tested positive for T. gondii antibodies. Conclusions This is the first prevalence study of T. gondii infection in Norwegian goats. The results show that Norwegian goat herds are commonly exposed to T. gondii. Nevertheless, the majority of goat herds have a low prevalence of antibody positive animals, which make them vulnerable to infections with T. gondii during the gestation period.

  7. Seroprevalence of Toxoplasma gondii infection in slaughtered pigs ...

    African Journals Online (AJOL)

    Toxoplasmosis is a parasitic disease/infection of medical and veterinary importance. The causative agent; Toxoplasma gondii, can infect warm blooded animals, birds as well as humans. This study was designed to determine the seroprevalence of Toxoplasma gondii infection in slaughtered pigs in Makurdi, Nigeria.

  8. Seroprevalence of Toxoplasma gondii and potential risk factors in ...

    African Journals Online (AJOL)

    Toxoplasma gondii infection is important in pigs and humans may get infected through the consumption of undercooked infected pork. This study conducted in Oyo state, Nigeria for 15 months (between February, 2012 and April, 2013) investigated the seroprevalence of Toxoplasma gondii infection in pigs reared on farms ...

  9. On the determination of Toxoplasma gondii virulence in mice

    Science.gov (United States)

    Toxoplasma gondii is one of the most successful pathogens on earth, capable of infecting mammals and birds. Numerous papers and reports are published on isolation of T .gondii from various natural sources worldwide. The house mouse (Mus musculus) has been used as the laboratory animal model to deter...

  10. Prevalence of Toxoplasma gondii in common moles (Talpa europaea)

    NARCIS (Netherlands)

    Krijger, I.M.; Cornelissen, J.B.W.J.; Wisselink, H.J.; Meerburg, B.G.

    2014-01-01

    Background The prevalence of Toxoplasma gondii in common moles, Talpa europaea, was investigated in order to determine whether moles can serve as an indicator species for T. gondii infections in livestock. Findings In total, 86 moles were caught from 25 different sites in the Netherlands. Five

  11. Seroprevalence of Toxoplasma gondii and Neospora caninum in ...

    African Journals Online (AJOL)

    Background: Toxoplasma gondii and Neospora caninum are protozoans infecting a wide range of mammals; the etiologic agents of Toxoplasmosis and Neosporosis respectively, This study investigated the prevalence of antibodies to Toxoplasma gondii and Neospora caninum in dogs from southwestern Nigeria. Materials ...

  12. Increased apoptosis skull of pups born to Toxoplasma gondii ...

    African Journals Online (AJOL)

    Background: Toxoplasma gondii is an intracellular obligate protozoan parasite that infects most warm-blooded animals including humans. It can cause congenital infection with clinical symptoms ranging from mild to severe including microcephaly. At the cellular level, infection T. gondii causes apoptosis in some tissues and ...

  13. Toxoplasma gondii seroprevalence in dairy and beef cattle

    DEFF Research Database (Denmark)

    Jokelainen, Pikka; Tagel, Maarja; Motus, Kerli

    2017-01-01

    Toxoplasma gondii is a zoonotic protozoan parasite that thrives in Estonia. In this nationwide cross-sectional study, we tested sera from 3991 cattle, collected from 228 farms in 2012–2013, for anti-T. gondii immunoglobulin G antibodies using a commercial direct agglutination test. Titer of 100 w...

  14. Biochemical characterization of the prolyl 3-hydroxylase 1.cartilage-associated protein.cyclophilin B complex.

    Science.gov (United States)

    Ishikawa, Yoshihiro; Wirz, Jackie; Vranka, Janice A; Nagata, Kazuhiro; Bächinger, Hans Peter

    2009-06-26

    The rough endoplasmic reticulum-resident protein complex consisting of prolyl 3-hydroxylase 1 (P3H1), cartilage-associated protein (CRTAP), and cyclophilin B (CypB) can be isolated from chick embryos on a gelatin-Sepharose column, indicating some involvement in the biosynthesis of procollagens. Prolyl 3-hydroxylase 1 modifies a single proline residue in the alpha chains of type I, II, and III collagens to (3S)-hydroxyproline. The peptidyl-prolyl cis-trans isomerase activity of cyclophilin B was shown previously to catalyze the rate of triple helix formation. Here we show that cyclophilin B in the complex shows peptidyl-prolyl cis-trans isomerase activity and that the P3H1.CRTAP.CypB complex has another important function: it acts as a chaperone molecule when tested with two classical chaperone assays. The P3H1.CRTAP.CypB complex inhibited the thermal aggregation of citrate synthase and was active in the denatured rhodanese refolding and aggregation assay. The chaperone activity of the complex was higher than that of protein-disulfide isomerase, a well characterized chaperone. The P3H1.CRTAP.CypB complex also delayed the in vitro fibril formation of type I collagen, indicating that this complex is also able to interact with triple helical collagen and acts as a collagen chaperone.

  15. Cyclophilin B stimulates RNA synthesis by the HCV RNA dependent RNA polymerase.

    Science.gov (United States)

    Heck, Julie A; Meng, Xiao; Frick, David N

    2009-04-01

    Cyclophilins are cellular peptidyl isomerases that have been implicated in regulating hepatitis C virus (HCV) replication. Cyclophilin B (CypB) is a target of cyclosporin A (CsA), an immunosuppressive drug recently shown to suppress HCV replication in cell culture. Watashi et al. recently demonstrated that CypB is important for efficient HCV replication, and proposed that it mediates the anti-HCV effects of CsA through an interaction with NS5B [Watashi K, Ishii N, Hijikata M, Inoue D, Murata T, Miyanari Y, et al. Cyclophilin B is a functional regulator of hepatitis C virus RNA polymerase. Mol Cell 2005;19:111-22]. We examined the effects of purified CypB proteins on the enzymatic activity of NS5B. Recombinant CypB purified from insect cells directly stimulated NS5B-catalyzed RNA synthesis. CypB increased RNA synthesis by NS5B derived from genotype 1a, 1b, and 2a HCV strains. Stimulation appears to arise from an increase in productive RNA binding. NS5B residue Pro540, a previously proposed target of CypB peptidyl-prolyl isomerase activity, is not required for stimulation of RNA synthesis.

  16. Genetic diversity of Toxoplasma gondii isolates from Ethiopian feral cats.

    Science.gov (United States)

    Dubey, J P; Choudhary, S; Tilahun, G; Tiao, N; Gebreyes, W A; Zou, X; Su, C

    2013-09-01

    Recent studies indicate greater genetic variability among isolates of Toxoplasma gondii worldwide than previously thought. However, there is no information on genetic diversity of T. gondii from any host in Ethiopia. In the present study, genotyping was performed on viable T. gondii isolates by bioassays in mice from tissues and feces of 27 cats from Ethiopia. Viable T. gondii was isolated from hearts of 26 cats, feces alone of 1 cat, and feces and tissues of 6 cats; in total there were 33 isolates. Genotyping was performed on DNA from cell-cultured derived T. gondii tachyzoites and by using 10 PCR-restriction fragment length polymorphism markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico). Four genotypes were recognized, including ToxoDB #1 (Type II clonal, nine isolates), ToxoDB #2 (Type III, five isolates), Toxo DB #3 (Type II variant, ten isolates), and ToxoDB #20 (nine isolates). Of interest is the isolation of different genotypes from tissues and feces of two cats, suggesting re-infection or mixed strain T. gondii infection. These findings are of epidemiological significance with respect to shedding of oocysts by cats. This is the first report of genotyping of T. gondii from any host in Ethiopia. Published by Elsevier B.V.

  17. Isolation and Genotyping of Toxoplasma gondii in Brazilian Dogs.

    Science.gov (United States)

    da Silva, Jamille Rodrigues; Maciel, Bianca Mendes; de Santana Souza Santos, Luana Karla Nogueira; Carvalho, Fábio Santos; de Santana Rocha, Daniele; Lopes, Carlos Wilson Gomes; Albuquerque, George Rêgo

    2017-06-01

    Strains of Toxoplasma gondii in Brazil are highly genetically diverse compared to strains from North America and Europe. Dogs are epidemiologically important because they act as sentinels for T. gondii infections in humans and are good indicators of environmental contamination. The aim of this study was to isolate and genetically characterize T. gondii strains from tissues of naturally infected Brazilian dogs. For this study, 21 blood samples were collected from dogs at the Zoonosis Control Centers of Ilhéus and Itabuna cities, Bahia, Brazil. The sera were examined for T. gondii antibodies using the indirect hemagglutination test. Brains and hearts of seropositive dogs were bioassayed in mice to isolate and characterize T. gondii parasites by PCR-RFLP using 10 genetic markers (SAG1, newSAG2, SAG3, BTUB, c22-8, c29-2, GRA6, PK1, APICO, and L358). However, T. gondii was isolated from only 4 (57.1%) dogs, designated TgDgBr6, 13, 17, and 21. All strains were virulent, causing clinical changes (rough hair coat, lethargy, and abdominal distention) and the death of all mice within 8-20 days after inoculation. Genetic analysis of these 4 T. gondii isolates revealed 4 distinct genotypes with different clonal lineage combinations (types I, II, and III) and 2 atypical alleles. Using PCR-RFLP with several markers, this study contributes to evaluations of the genetic diversity of strains circulating in Brazil.

  18. Toxoplasma gondii antibodies in the white stork Ciconia ciconia.

    Science.gov (United States)

    Andrzejewska, Izabela; Tryjanowski, Piotr; Zduniak, Piotr; Dolata, Pawel T; Ptaszyk, Jerzy; Cwiertnia, Piotr

    2004-01-01

    The prevalence of Toxoplasma gondii in chicks of wild birds and captive individuals was studied in the Poznań environs and in the Poznań Zoological Garden in the years 2002-2003. Bird blood was tested for T. gondii antibodies by an indirect fluorescent antibody test. T. gondii antibodies were detected from 5.8% of 205 analysed white stork chicks and 13.6% of 44 analysed adult storks in the zoo. Because toxoplasmosis is one of the more common parasitic zoonoses worldwide, we briefly discuss the potential epidemiological importance of stork toxoplasmosis to humans.

  19. Extracellular cyclophilin levels associate with parameters of asthma in phenotypic clusters.

    Science.gov (United States)

    Stemmy, Erik J; Benton, Angela S; Lerner, Jennifer; Alcala, Sarah; Constant, Stephanie L; Freishtat, Robert J

    2011-12-01

    Leukocyte persistence during chronic (quiescent) phases of asthma is a major hallmark of the disease. The mechanisms regulating these persistent leukocyte populations are not clearly understood. An alternative family of chemoattracting proteins, cyclophilins (Cyps), has recently been shown to contribute to leukocyte recruitment in animal models of allergic asthma. The goals of this study were to determine whether Cyps are present in asthma patients during the chronic phase of the disease and to investigate whether levels of Cyps associate with clinical parameters of disease severity. Nasal wash samples from an urban cohort of 137 patients of age 6-20 years with physician-diagnosed asthma were examined for the presence of cyclophilin A (CypA), cyclophilin B (CypB), as well as several other classical chemokines. Linear, logistic, or ordinal regressions were performed to identify associations between Cyps, chemokines, and clinical parameters of asthma. The asthma cohort was further divided into previously established phenotypic clusters (cluster 1: n = 55; cluster 2: n = 31; and cluster 3: n = 51) and examined for associations. Levels of CypB in the asthma group were highly elevated compared to nonasthmatic controls, while a slight increase in Monocyte Chemotactic Protein-1 (MCP-1) was also observed. CypA and MCP-1 were associated with levels of eosinophil cationic protein (ECP; a marker of eosinophil activation). Cluster-specific associations were found for CypA and CypB and clinical asthma parameters [e.g. forced expiratory volume in 1 second (FEV(1)) and ECP]. Cyps are present in nasal wash samples of asthma patients and may be a novel biomarker for clinical parameters of asthma severity.

  20. Evidence that human milk isolated cyclophilin B corresponds to a truncated form.

    Science.gov (United States)

    Mariller, C; Allain, F; Kouach, M; Spik, G

    1996-03-07

    Cyclophilin B (CyPB) is a member of the cyclophilin family (cyclosporin A-binding proteins) with specific N- and C-terminal extensions. In contrast to cyclophilin A, CyPB owns a signal sequence leading to its translocation in the endoplasmic reticulum. CyPB was reported to be present in human blood and milk, suggesting it is secreted. For this purpose, CyPB was purified to homogeneity from human milk and compared to recombinant CyPB expressed in E. coli. Ion spray mass spectrometry revealed that CyPB secreted in human milk exhibits a lower molecular mass than the one expected. Identification of phenylalanine as the C-terminus amino-acid residue of human milk CyPB indicates that the difference in molecular mass may be explained by the absence of the five C-terminal amino-acid residues AIAKE. These results suggest that in the sequence VEKPFAIAKE known to be responsible for retention of CyPB in the endoplasmic reticulum, the sequence AIAKE is more particularly necessary. Our findings raise the possibility that the CyPB may be processed to promote its release. As recombinant CyPB was shown to bind specifically to Jurkat cells, a lymphoblastic T-cell line, we then wanted to investigate the binding of human milk CyPB to these cells. Despite lacking the five C-terminal amino-acid residues, human milk CyPB is able to inhibit the binding of recombinant CyPB to Jurkat T cells.

  1. Fatal Toxoplasma gondii infection in the giant panda.

    Science.gov (United States)

    Ma, Hongyu; Wang, Zedong; Wang, Chengdong; Li, Caiwu; Wei, Feng; Liu, Quan

    2015-01-01

    Toxoplasma gondii can infect nearly all warm-blooded animals. We report an acute fatal T. gondii infection in the endangered giant panda (Ailuropoda melanoleuca) in a zoo in China, characterized by acute gastroenteritis and respiratory symptoms. T. gondii infection was confirmed by immunological and molecular methods. Multilocus nested PCR-RFLP revealed clonal type I at the SAG1 and c29-2 loci, clonal type II at the SAG2, BTUB, GRA6, c22-8, and L358 loci, and clonal type III at the alternative SAG2 and SAG3 loci, thus, a potential new genotype of T. gondii in the giant panda. Other possible pathogens were not detected. To our knowledge, this is the first report of clinical toxoplasmosis in a giant panda. © H. Ma et al., published by EDP Sciences, 2015.

  2. Fatal Toxoplasma gondii infection in the giant panda

    Directory of Open Access Journals (Sweden)

    2015-01-01

    Full Text Available Toxoplasma gondii can infect nearly all warm-blooded animals. We report an acute fatal T. gondii infection in the endangered giant panda (Ailuropoda melanoleuca in a zoo in China, characterized by acute gastroenteritis and respiratory symptoms. T. gondii infection was confirmed by immunological and molecular methods. Multilocus nested PCR-RFLP revealed clonal type I at the SAG1 and c29-2 loci, clonal type II at the SAG2, BTUB, GRA6, c22-8, and L358 loci, and clonal type III at the alternative SAG2 and SAG3 loci, thus, a potential new genotype of T. gondii in the giant panda. Other possible pathogens were not detected. To our knowledge, this is the first report of clinical toxoplasmosis in a giant panda.

  3. Outbreak of caprine abortion by Toxoplasma gondii in Midwest Brazil

    Directory of Open Access Journals (Sweden)

    Flávio Henrique Bravim Caldeira

    2011-11-01

    Full Text Available An outbreak of abortion by Toxoplasma gondii in goats on a farm in the Brazilian Midwest is reported. Gross lesions were not observed in seven aborted fetuses submitted to the Veterinary Pathology Laboratory, Federal University of Mato Grosso, for necropsy investigation. The main histologic lesions were mononuclear cell pneumonia and necrotizing encephalitis in varying degrees of intensity. PCR for Brucella abortus and Neospora caninum and aerobic cultures were negative in all cases. Antibody titles against T. gondii varying from 1:1024 to 1:32.768 were detected in serum samples from four aborted goats. Nested-PCR assay for T. gondii were positive in brain samples of all cases submitted. These findings indicate that T. gondii infection should be considered in the diagnosis of abortion in goats in Midwest Brazil.

  4. Experimental toxoplasma gondii infection in grey seals (Halichoerus grypus)

    DEFF Research Database (Denmark)

    Gajadhar, A. A.; Measures, L.; Forbes, L. B.

    2004-01-01

    Laboratory-reared animals were used to assess the susceptibility of seals (Halichoerus grypus) to Toxoplasma gondii infection. Four seals were each orally inoculated with 100 or 10,000 oocysts of T. gondii (VEG strain), and another 4 seals served as negative controls. Occasionally, mild behavioral...... changes were observed in all inoculated seals but not in control animals. A modified agglutination test revealed the presence of antibodies to T. gondii in sera collected from inoculated seals and mice inoculated as controls. No evidence of the parasite was found on an extensive histological examination...... of seal tissues, and immunohistochemical staining of tissue sections from inoculated seals revealed a single tissue cyst in only 1 seal. Control mice inoculated with 10 oocysts from the same inoculum given to seals became serologically and histologically positive for T. gondii. Cats that were fed brain...

  5. Elevated Serum Cyclophilin B Levels Are Associated with the Prevalence and Severity of Metabolic Syndrome

    OpenAIRE

    Zhang, Hang; Fan, Qin; Xie, Hongyang; Lu, Lin; Tao, Rong; Wang, Fang; Xi, Rui; Hu, Jian; Chen, Qiujing; Shen, Weifeng; Zhang, Ruiyan; Yan, Xiaoxiang

    2017-01-01

    Objective Inflammation plays a central role in the pathogenesis of metabolic syndrome (MetS). Cyclophilin B (CypB) can be constitutively secreted in response to inflammatory stimuli and oxidative stress, participating in tissue or systemic inflammation. We investigated the relationship between CypB and MetS in both humans and mice. Methods Serum CypB levels were determined in 211 subjects with MetS and 292 subjects without MetS (non-MetS) (133 healthy controls and 159 high-risk subjects with ...

  6. Elevated Serum Cyclophilin B Levels Are Associated with the Prevalence and Severity of Metabolic Syndrome

    OpenAIRE

    Hang Zhang; Hang Zhang; Qin Fan; Qin Fan; Hongyang Xie; Hongyang Xie; Lin Lu; Lin Lu; Rong Tao; Fang Wang; Rui Xi; Jian Hu; Qiujing Chen; Weifeng Shen; Ruiyan Zhang

    2017-01-01

    ObjectiveInflammation plays a central role in the pathogenesis of metabolic syndrome (MetS). Cyclophilin B (CypB) can be constitutively secreted in response to inflammatory stimuli and oxidative stress, participating in tissue or systemic inflammation. We investigated the relationship between CypB and MetS in both humans and mice.MethodsSerum CypB levels were determined in 211 subjects with MetS and 292 subjects without MetS (non-MetS) (133 healthy controls and 159 high-risk subjects with one...

  7. Mitochondrial cyclophilin-D as a critical mediator of ischaemic preconditioning

    OpenAIRE

    Hausenloy, Derek J.; Lim, Shiang Y.; Ong, Sang-Ging; Davidson, Sean M.; Yellon, Derek M.

    2010-01-01

    Aims It has been suggested that mitochondrial reactive oxygen species (ROS), Akt and Erk1/2 and more recently the mitochondrial permeability transition pore (mPTP) may act as mediators of ischaemic preconditioning (IPC), although the actual interplay between these mediators is unclear. The aim of the present study is to determine whether the cyclophilin-D (CYPD) component of the mPTP is required by IPC to generate mitochondrial ROS and subsequently activate Akt and Erk1/2. Methods and results...

  8. Adenosine metabolism in Toxoplasma gondii: potential targets for chemotherapy.

    Science.gov (United States)

    el Kouni, Mahmoud H

    2007-01-01

    Toxoplasma gondii is an intracellular parasitic protozoan that infects approximately a billion people worldwide. Infection with T. gondii represents a major health problem for immunocompromised individuals, such as AIDS patients, organ transplant recipients, and the unborn children of infected mothers. Currently available drugs usually do not eradicate infection and as many as 50% of the patients do not respond to this therapy. Furthermore, they are ineffective against T. gondii tissue cysts. In addition, prolonged exposure to these drugs induces serious host toxicity forcing the discontinuation of the therapy. Finally, there is no effective vaccine currently available for the treatment of toxoplasmosis. Therefore, it is necessary to develop new and effective drugs for the treatment and management of toxoplasmosis. The rational design of a drug depends on the exploitation of fundamental biochemical or physiological differences between pathogens and their host. Some of the most striking differences between T. gondii and their mammalian host are found in purine metabolism. T. gondii, like most parasites studied, lack the ability to synthesize purines do novo and depend on the salvage of purines from their host to satisfy their requirements of purines. In this respect, the salvage of adenosine is the major source of purines in T. gondii. Therefore, interference with adenosine uptake and metabolism in T. gondii can be selectively detrimental to the parasite. The host cells, on the other hand, can still obtain their purine requirements by their de novo pathways. This review will focus on the broad aspects of the adenosine transport and the enzyme adenosine kinase (EC 2.7.1.20) which are the two primary routes for adenosine utilization in T. gondii, in an attempt to illustrate their potentials as targets for chemotherapy against this parasite.

  9. Mutation in cyclophilin B that causes hyperelastosis cutis in American Quarter Horse does not affect peptidylprolyl cis-trans isomerase activity but shows altered cyclophilin B-protein interactions and affects collagen folding.

    Science.gov (United States)

    Ishikawa, Yoshihiro; Vranka, Janice A; Boudko, Sergei P; Pokidysheva, Elena; Mizuno, Kazunori; Zientek, Keith; Keene, Douglas R; Rashmir-Raven, Ann M; Nagata, Kazuhiro; Winand, Nena J; Bächinger, Hans Peter

    2012-06-22

    The rate-limiting step of folding of the collagen triple helix is catalyzed by cyclophilin B (CypB). The G6R mutation in cyclophilin B found in the American Quarter Horse leads to autosomal recessive hyperelastosis cutis, also known as hereditary equine regional dermal asthenia. The mutant protein shows small structural changes in the region of the mutation at the side opposite the catalytic domain of CypB. The peptidylprolyl cis-trans isomerase activity of the mutant CypB is normal when analyzed in vitro. However, the biosynthesis of type I collagen in affected horse fibroblasts shows a delay in folding and secretion and a decrease in hydroxylysine and glucosyl-galactosyl hydroxylysine. This leads to changes in the structure of collagen fibrils in tendon, similar to those observed in P3H1 null mice. In contrast to cyclophilin B null mice, where little 3-hydroxylation was found in type I collagen, 3-hydroxylation of type I collagen in affected horses is normal. The mutation disrupts the interaction of cyclophilin B with the P-domain of calreticulin, with lysyl hydroxylase 1, and probably other proteins, such as the formation of the P3H1·CypB·cartilage-associated protein complex, resulting in less effective catalysis of the rate-limiting step in collagen folding in the rough endoplasmic reticulum.

  10. Mutation in Cyclophilin B That Causes Hyperelastosis Cutis in American Quarter Horse Does Not Affect Peptidylprolyl cis-trans Isomerase Activity but Shows Altered Cyclophilin B-Protein Interactions and Affects Collagen Folding*

    Science.gov (United States)

    Ishikawa, Yoshihiro; Vranka, Janice A.; Boudko, Sergei P.; Pokidysheva, Elena; Mizuno, Kazunori; Zientek, Keith; Keene, Douglas R.; Rashmir-Raven, Ann M.; Nagata, Kazuhiro; Winand, Nena J.; Bächinger, Hans Peter

    2012-01-01

    The rate-limiting step of folding of the collagen triple helix is catalyzed by cyclophilin B (CypB). The G6R mutation in cyclophilin B found in the American Quarter Horse leads to autosomal recessive hyperelastosis cutis, also known as hereditary equine regional dermal asthenia. The mutant protein shows small structural changes in the region of the mutation at the side opposite the catalytic domain of CypB. The peptidylprolyl cis-trans isomerase activity of the mutant CypB is normal when analyzed in vitro. However, the biosynthesis of type I collagen in affected horse fibroblasts shows a delay in folding and secretion and a decrease in hydroxylysine and glucosyl-galactosyl hydroxylysine. This leads to changes in the structure of collagen fibrils in tendon, similar to those observed in P3H1 null mice. In contrast to cyclophilin B null mice, where little 3-hydroxylation was found in type I collagen, 3-hydroxylation of type I collagen in affected horses is normal. The mutation disrupts the interaction of cyclophilin B with the P-domain of calreticulin, with lysyl hydroxylase 1, and probably other proteins, such as the formation of the P3H1·CypB·cartilage-associated protein complex, resulting in less effective catalysis of the rate-limiting step in collagen folding in the rough endoplasmic reticulum. PMID:22556420

  11. Toxoplasma gondii in wild and domestic animals from New Caledonia

    Directory of Open Access Journals (Sweden)

    Roqueplo C.

    2011-11-01

    Full Text Available Samples (serum or meat juice collected from 205 animals in New Caledonia in April 2009 were tested for antibodies against Toxoplasma gondii by ELISA using the multi-species ID Screen® Toxoplasmosis Indirect kit (IDVET, Montpellier. Antibodies to T. gondii were detected in 2% (1/49 of the pigs, in 3.3% (1/30 of the cattle, in 13.8% (4/29 of Rusa deers, in 16% (4/25 of the horses, in 32.8% (21/64 of the dogs, and in 50% (4/8 of cats. Statistically, no significant difference was observed between T. gondii seroprevalence and age or sex. No survey on the prevalence of T. gondii in animals has ever been conducted in New Caledonia and this is the first serological evidence of T. gondii in Rusa deer (Cervus timorensis russa. These results indicate an important circulation of T. gondii exists in the animal populations of New Caledonia. In view of humans being exposed, it is advisable to insist on sanitary education and on respect for good hygienic and food practice.

  12. Toxoplasma gondii infection and chronic schizophrenia: is there any association?

    Directory of Open Access Journals (Sweden)

    Salvina Maria de Campos-Carli

    Full Text Available ABSTRACT Background: Toxoplasma gondii (T. gondii infection has been identified as a risk factor for schizophrenia. Objectives: Herein, we sought to evaluate the association between T. gondii infection and clinical symptoms and quality of life in patients with schizophrenia. Methods: We conducted a cross-sectional study with 48 patients with chronic schizophrenia and 40 controls. Peripheral blood was drawn, and IgM and IgG anti-T. gondii antibodies were evaluated by Enzyme-Linked Immunosorbent Assay (ELISA. Depressive, positive and negative symptoms were assessed, respectively, by the Calgary Depression Scale (CDS and the Positive and Negative Syndrome Scale (PANSS. Cognitive performance was assessed in patients by the Brazilian version of the Schizophrenia Cognition Rating Scale (SCoRS-BR. Quality of life was assessed by the Brazilian version of the Quality of Life in Schizophrenia scale (QLS-BR. Results: The prevalence and titers of T. gondii IgM and IgG antibodies did not differ between patients and controls. The positive serology for T. gondii IgG antibodies was not associated with illness symptoms, cognitive performance, depressive symptoms or quality of life. Discussion: Our findings suggest that toxoplasmosis infection is not associated with severity of symptoms, quality of life, cognitive or depressive symptoms in schizophrenia patients.

  13. Diagnosis of toxoplasmosis and typing of Toxoplasma gondii.

    Science.gov (United States)

    Liu, Quan; Wang, Ze-Dong; Huang, Si-Yang; Zhu, Xing-Quan

    2015-05-28

    Toxoplasmosis, caused by the obligate intracellular protozoan Toxoplasma gondii, is an important zoonosis with medical and veterinary importance worldwide. The disease is mainly contracted by ingesting undercooked or raw meat containing viable tissue cysts, or by ingesting food or water contaminated with oocysts. The diagnosis and genetic characterization of T. gondii infection is crucial for the surveillance, prevention and control of toxoplasmosis. Traditional approaches for the diagnosis of toxoplasmosis include etiological, immunological and imaging techniques. Diagnosis of toxoplasmosis has been improved by the emergence of molecular technologies to amplify parasite nucleic acids. Among these, polymerase chain reaction (PCR)-based molecular techniques have been useful for the genetic characterization of T. gondii. Serotyping methods based on polymorphic polypeptides have the potential to become the choice for typing T. gondii in humans and animals. In this review, we summarize conventional non-DNA-based diagnostic methods, and the DNA-based molecular techniques for the diagnosis and genetic characterization of T. gondii. These techniques have provided foundations for further development of more effective and accurate detection of T. gondii infection. These advances will contribute to an improved understanding of the epidemiology, prevention and control of toxoplasmosis.

  14. Synthesis of heparan sulfate with cyclophilin B-binding properties is determined by cell type-specific expression of sulfotransferases.

    NARCIS (Netherlands)

    Deligny, A.; Denys, A.; Marcant, A.; Melchior, A.; Mazurier, J.; Kuppevelt, A.H.M.S.M. van; Allain, F.

    2010-01-01

    Cyclophilin B (CyPB) induces migration and adhesion of T lymphocytes via a mechanism that requires interaction with 3-O-sulfated heparan sulfate (HS). HS biosynthesis is a complex process with many sulfotransferases involved. N-Deacetylases/N-sulfotransferases are responsible for N-sulfation, which

  15. IMPDH2 Is an Intracellular Target of the Cyclophilin A and Sanglifehrin A Complex

    Directory of Open Access Journals (Sweden)

    Khian Hong Pua

    2017-01-01

    Full Text Available Summary: Natural products have demonstrated utility in the clinic and can also act as probes to understand complex cellular pathways. Sanglifehrin A (SFA is a mixed polyketide and non-ribosomal peptide synthase natural product with sub-nano-molar affinity for its receptor cyclophilin A (PPIA. It has been shown to behave in vitro as an immune suppressant. Here, we identify inosine-5′-monophosphate dehydrogenase 2 (IMPDH2 as an intracellular target of the PPIA-SFA binary complex. The formation of this ternary complex does not inhibit the enzymatic activity of IMPDH2. Rather, ternary complex formation modulates cell growth through interaction with the cystathionine-β-synthase (CBS domain of IMPDH2. We further demonstrate that the SFA complex is highly isoform selective for IMPDH2 (versus IMPDH1. This work reveals a role for the CBS domains of IMPDH2 in cellular proliferation, suggesting a more complex role than previously suspected for IMPDH2 in T cell activation and proliferation. : Pua et al. identify IMPDH2 as an intracellular target of the PPIA-SFA complex and show that the CBS domains of IMPDH2 are required for cellular proliferation. Keywords: cyclophilin A, sanglifehrin A, inosine-5′-monophosphate dehydrogenase, cystathionine-β-synthase domains, protein-protein interactions

  16. The level of CD147 expression correlates with cyclophilin-induced signalling and chemotaxis

    Directory of Open Access Journals (Sweden)

    Constant Stephanie

    2011-10-01

    Full Text Available Abstract Background Previous studies identified CD147 as the chemotactic receptor on inflammatory leukocytes for extracellular cyclophilins (eCyp. However, CD147 is not known to associate with signal transducing molecules, so other transmembrane proteins, such as proteoglycans, integrins, and CD98, were suggested as receptor or co-receptor for eCyp. CD147 is ubiquitously expressed on many cell types, but relationship between the level of CD147 expression and cellular responses to eCyp has never been analyzed. Given the role of eCyp in pathogenesis of many diseases, it is important to know whether cellular responses to eCyp are regulated at the level of CD147 expression. Results Here, we manipulated CD147 expression levels on HeLa cells using RNAi and investigated the signalling and chemotactic responses to eCypA. Both Erk activation and chemotaxis correlated with the level of CD147 expression, with cells exhibiting low level expression being practically unresponsive to eCypA. Conclusions Our results provide the first demonstration of a chemotactic response of HeLa cells to eCypA, establish a correlation between the level of CD147 expression and the magnitude of cellular responses to eCypA, and indicate that CD147 may be a limiting factor in the receptor complex determining cyclophilin-induced Erk activation and cell migration.

  17. Cyclophilin D links programmed cell death and organismal aging in Podospora anserina.

    Science.gov (United States)

    Brust, Diana; Daum, Bertram; Breunig, Christine; Hamann, Andrea; Kühlbrandt, Werner; Osiewacz, Heinz D

    2010-10-01

    Cyclophilin D (CYPD) is a mitochondrial peptidyl prolyl-cis,trans-isomerase involved in opening of the mitochondrial permeability transition pore (mPTP). CYPD abundance increases during aging in mammalian tissues and in the aging model organism Podospora anserina. Here, we show that treatment of the P. anserina wild-type with low concentrations of the cyclophilin inhibitor cyclosporin A (CSA) extends lifespan. Transgenic strains overexpressing PaCypD are characterized by reduced stress tolerance, suffer from pronounced mitochondrial dysfunction and are characterized by accelerated aging and induction of cell death. Treatment with CSA leads to correction of mitochondrial function and lifespan to that of the wild-type. In contrast, PaCypD deletion strains are not affected by CSA within the investigated concentration range and show increased resistance against inducers of oxidative stress and cell death. Our data provide a mechanistic link between programmed cell death (PCD) and organismal aging and bear implications for the potential use of CSA to intervene into biologic aging. © 2010 The Authors Aging Cell © 2010 Blackwell Publishing Ltd/Anatomical Society of Great Britain and Ireland.

  18. Preclinical characterization of naturally occurring polyketide cyclophilin inhibitors from the sanglifehrin family.

    Science.gov (United States)

    Gregory, Matthew A; Bobardt, Michael; Obeid, Susan; Chatterji, Udayan; Coates, Nigel J; Foster, Teresa; Gallay, Philippe; Leyssen, Pieter; Moss, Steven J; Neyts, Johan; Nur-e-Alam, Mohammad; Paeshuyse, Jan; Piraee, Mahmood; Suthar, Dipen; Warneck, Tony; Zhang, Ming-Qiang; Wilkinson, Barrie

    2011-05-01

    Cyclophilin inhibitors currently in clinical trials for hepatitis C virus (HCV) are all analogues of cyclosporine (CsA). Sanglifehrins are a group of naturally occurring cyclophilin binding polyketides that are structurally distinct from the cyclosporines and are produced by a microorganism amenable to biosynthetic engineering for lead optimization and large-scale production by fermentation. Preclinical characterization of the potential utility of this class of compounds for the treatment of HCV revealed that the natural sanglifehrins A to D are all more potent than CsA at disrupting formation of the NS5A-CypA, -CypB, and -CypD complexes and at inhibition of CypA, CypB, and CypD isomerase activity. In particular, sanglifehrin B (SfB) was 30- to 50-fold more potent at inhibiting the isomerase activity of all Cyps tested than CsA and was also shown to be a more potent inhibitor of the 1b subgenomic replicon (50% effective concentrations [EC50s] of 0.070 μM and 0.16 μM in Huh 5-2 and Huh 9-13 cells, respectively). Physicochemical and mouse pharmacokinetic analyses revealed low oral bioavailability (F5 h). These data demonstrate that naturally occurring sanglifehrins are suitable lead compounds for the development of novel analogues that are less immunosuppressive and that have improved metabolism and pharmacokinetic properties.

  19. Preclinical Characterization of Naturally Occurring Polyketide Cyclophilin Inhibitors from the Sanglifehrin Family▿†

    Science.gov (United States)

    Gregory, Matthew A.; Bobardt, Michael; Obeid, Susan; Chatterji, Udayan; Coates, Nigel J.; Foster, Teresa; Gallay, Philippe; Leyssen, Pieter; Moss, Steven J.; Neyts, Johan; Nur-e-Alam, Mohammad; Paeshuyse, Jan; Piraee, Mahmood; Suthar, Dipen; Warneck, Tony; Zhang, Ming-Qiang; Wilkinson, Barrie

    2011-01-01

    Cyclophilin inhibitors currently in clinical trials for hepatitis C virus (HCV) are all analogues of cyclosporine (CsA). Sanglifehrins are a group of naturally occurring cyclophilin binding polyketides that are structurally distinct from the cyclosporines and are produced by a microorganism amenable to biosynthetic engineering for lead optimization and large-scale production by fermentation. Preclinical characterization of the potential utility of this class of compounds for the treatment of HCV revealed that the natural sanglifehrins A to D are all more potent than CsA at disrupting formation of the NS5A-CypA, -CypB, and -CypD complexes and at inhibition of CypA, CypB, and CypD isomerase activity. In particular, sanglifehrin B (SfB) was 30- to 50-fold more potent at inhibiting the isomerase activity of all Cyps tested than CsA and was also shown to be a more potent inhibitor of the 1b subgenomic replicon (50% effective concentrations [EC50s] of 0.070 μM and 0.16 μM in Huh 5-2 and Huh 9-13 cells, respectively). Physicochemical and mouse pharmacokinetic analyses revealed low oral bioavailability (F 5 h). These data demonstrate that naturally occurring sanglifehrins are suitable lead compounds for the development of novel analogues that are less immunosuppressive and that have improved metabolism and pharmacokinetic properties. PMID:21383094

  20. Cyclophilin B Supports Myc and Mutant p53 Dependent Survival of Glioblastoma Multiforme Cells

    Science.gov (United States)

    Choi, Jae Won; Schroeder, Mark A.; Sarkaria, Jann N.; Bram, Richard J.

    2014-01-01

    Glioblastoma multiforme (GBM) is an aggressive, treatment-refractory type of brain tumor for which effective therapeutic targets remain important to identify. Here we report that cyclophilin B (CypB), a prolyl isomerase residing in the endoplasmic reticulum (ER), provides an essential survival signal in GBM cells. Analysis of gene expression databases revealed that CypB is upregulated in many cases of malignant glioma. We found that suppression of CypB reduced cell proliferation and survival in human GBM cells in vitro and in vivo. We also found that treatment with small molecule inhibitors of cyclophilins, including the approved drug cyclosporine, greatly reduced the viability of GBM cells. Mechanistically, depletion or pharmacologic inhibition of CypB caused hyperactivation of the oncogenic RAS-MAPK pathway, induction of cellular senescence signals, and death resulting from loss of MYC, mutant p53, Chk1 and JAK/STAT3 signaling. Elevated reactive oxygen species, ER expansion and abnormal unfolded protein responses in CypB-depleted GBM cells indicated that CypB alleviates oxidative and ER stresses and coordinates stress adaptation responses. Enhanced cell survival and sustained expression of multiple oncogenic proteins downstream of CypB may thus contribute to the poor outcome of GBM tumors. Our findings link chaperone-mediated protein folding in the ER to mechanisms underlying oncogenic transformation, and they make CypB an attractive and immediately targetable molecule for GBM therapy. PMID:24272483

  1. First characterization of three cyclophilin family proteins in the oyster, Crassostrea ariakensis Gould.

    Science.gov (United States)

    Xu, Ting; Xie, Jiasong; Yang, Shoubao; Ye, Shigen; Luo, Ming; Wu, Xinzhong

    2016-08-01

    Cyclophilins (CyPs) are a family of proteins that bind the immunosuppressive agent cyclosporin A (CsA) with high-affinity and belong to one of the three superfamilies of peptidyl-prolyl cis-trans isomerases (PPIase). In this report, three cyclophilin genes (Ca-CyPs), including Ca-CyPA, Ca-CyPB and Ca-PPIL3, were identified from oyster, Crassostrea ariakensis Gould in which Ca-CyPA encodes a protein with 165 amino acid sequences, Ca-CyPB encodes a protein with 217 amino acid sequences and Ca-PPIL3 encodes a protein with 162 amino acid sequences. All of the three Ca-CyPs genes contain a typical CyP-PPIase domain with its signature sequences and Ca-CyPB contains an N-signal peptide sequences. Tissue distribution study revealed that Ca-CyPs were ubiquitously expressed in all examined tissues and the highest levels were observed in hemocytes. RLO incubation upregulated the mRNA expression levels of Ca-CyPs, indicating that three Ca-CyPs might be involved in oyster immune response against RLO infection. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. Cyclophilin B supports Myc and mutant p53-dependent survival of glioblastoma multiforme cells.

    Science.gov (United States)

    Choi, Jae Won; Schroeder, Mark A; Sarkaria, Jann N; Bram, Richard J

    2014-01-15

    Glioblastoma multiforme is an aggressive, treatment-refractory type of brain tumor for which effective therapeutic targets remain important to identify. Here, we report that cyclophilin B (CypB), a prolyl isomerase residing in the endoplasmic reticulum (ER), provides an essential survival signal in glioblastoma multiforme cells. Analysis of gene expression databases revealed that CypB is upregulated in many cases of malignant glioma. We found that suppression of CypB reduced cell proliferation and survival in human glioblastoma multiforme cells in vitro and in vivo. We also found that treatment with small molecule inhibitors of cyclophilins, including the approved drug cyclosporine, greatly reduced the viability of glioblastoma multiforme cells. Mechanistically, depletion or pharmacologic inhibition of CypB caused hyperactivation of the oncogenic RAS-mitogen-activated protein kinase pathway, induction of cellular senescence signals, and death resulting from loss of MYC, mutant p53, Chk1, and Janus-activated kinase/STAT3 signaling. Elevated reactive oxygen species, ER expansion, and abnormal unfolded protein responses in CypB-depleted glioblastoma multiforme cells indicated that CypB alleviates oxidative and ER stresses and coordinates stress adaptation responses. Enhanced cell survival and sustained expression of multiple oncogenic proteins downstream of CypB may thus contribute to the poor outcome of glioblastoma multiforme tumors. Our findings link chaperone-mediated protein folding in the ER to mechanisms underlying oncogenic transformation, and they make CypB an attractive and immediately targetable molecule for glioblastoma multiforme therapy.

  3. Overexpressed cyclophilin B suppresses apoptosis associated with ROS and Ca2+ homeostasis after ER stress.

    Science.gov (United States)

    Kim, Jinhwan; Choi, Tae Gyu; Ding, Yan; Kim, Yeonghwan; Ha, Kwon Soo; Lee, Kyung Ho; Kang, Insug; Ha, Joohun; Kaufman, Randal J; Lee, Jinhwa; Choe, Wonchae; Kim, Sung Soo

    2008-11-01

    Prolonged accumulation of misfolded proteins in the endoplasmic reticulum (ER) results in ER stress-mediated apoptosis. Cyclophilins are protein chaperones that accelerate the rate of protein folding through their peptidyl-prolyl cis-trans isomerase (PPIase) activity. In this study, we demonstrated that ER stress activates the expression of the ER-localized cyclophilin B (CypB) gene through a novel ER stress response element. Overexpression of wild-type CypB attenuated ER stress-induced cell death, whereas overexpression of an isomerase activity-defective mutant, CypB/R62A, not only increased Ca(2+) leakage from the ER and ROS generation, but also decreased mitochondrial membrane potential, resulting in cell death following exposure to ER stress-inducing agents. siRNA-mediated inhibition of CypB expression rendered cells more vulnerable to ER stress. Finally, CypB interacted with the ER stress-related chaperones, Bip and Grp94. Taken together, we concluded that CypB performs a crucial function in protecting cells against ER stress via its PPIase activity.

  4. Toxoplasma gondii impairs memory in infected seniors.

    Science.gov (United States)

    Gajewski, Patrick D; Falkenstein, Michael; Hengstler, Jan G; Golka, Klaus

    2014-02-01

    Almost 30% of humans present a Toxoplasma gondii positive antibody status and its prevalence increases with age. The central nervous system is the main target. However, little is known about the influence of asymptomatic i.e. latent Toxoplasmosis on cognitive functions in humans. To investigate neurocognitive dysfunctions in asymptomatic older adults with T. gondii positive antibody status a double-blinded neuropsychological study was conducted. The participants were classified from a population-based sample (N=131) of healthy participants with an age of 65 years and older into two groups with 42 individuals each: Toxoplasmosis positive (T-pos; IgG>50 IU/ml) and Toxoplasmosis negative (T-neg; IgG=0 IU/ml). The outcome measures were a computer-based working-memory test (2-back) and several standardized psychometric tests of memory and executive cognitive functions. T-pos seniors showed an impairment of different aspects of memory. The rate of correctly detected target symbols in a 2-back task was decreased by nearly 9% (P=0.020), corresponding to a performance reduction of about 35% in working memory relative to the T-neg group. Moreover, T-pos seniors had a lower performance in a verbal memory test, both regarding immediate recall (10% reduction; P=0.022), delayed recognition (6%; P=0.037) and recall from long-term memory assessed by the word fluency tests (12%; P=0.029). In contrast, executive functions were not affected. The effects remained mostly unchanged after controlling for medication. The impairment of memory functions in T-pos seniors was accompanied by a decreased self-reported quality of life. Because of the high prevalence of asymptomatic Toxoplasmosis and an increasing population of older adults this finding is of high relevance for public health. Copyright © 2013 Elsevier Inc. All rights reserved.

  5. High prevalence of Toxoplasma gondii antibodies in dogs in Veracruz, Mexico

    Science.gov (United States)

    Little is known concerning the prevalence of Toxoplasma gondii infection in dogs in Mexico. Here, we investigated antibodies to T. gondii and associated risk factors in 101 dogs from an animal shelter in Veracruz State, Mexico. Canine sera were assayed for T. gondii IgG antibodies by using the modif...

  6. Use of filter papers to determine seroprevalence of Toxoplasma gondii among hunted ungulates

    Science.gov (United States)

    Toxoplasmosis is a zoonosis caused by the protozoan Toxoplasma gondii, and it is found worldwide. To determine whether ungulates are reservoirs of T. gondii in an isolated and remote region of the northeastern Peruvian Amazon, antibodies to T. gondii were determined in 5 species of ungulates by the...

  7. Serotyping of Toxoplasma gondii in Cats (Felis domesticus) Reveals Predominance of Type II Infections in Germany

    Science.gov (United States)

    Background: Cats are definitive hosts of Toxoplasma gondii and play an essential role in the epidemiology of this parasite. The study aims at clarifying whether cats are able to develop specific antibodies against different clonal types of T. gondii and to determine by serotyping the T. gondii clona...

  8. Isolation and genetic characterization of Toxoplasma gondii from the gray wolf Canis lupus

    Science.gov (United States)

    Little is known of the genetic diversity of Toxoplasma gondii circulating in wildlife. In the present study feral gray wolf (Canis lupus) from Minnesota were examined for T. gondii infection. Antibodies to T. gondii were detected in 130 (52.4%) of 248 wolves tested by the modified agglutination test...

  9. Seroprevalence and risk factors for Toxoplasma gondii infection in domestic cats in The Netherlands

    NARCIS (Netherlands)

    Opsteegh, M.; Haveman, R.; Swart, A.; Mensink-Beerepoot, M.E.; Hofhuis, A.; Langelaar, M.F.M.; van der Giessen, J.W.B.

    2012-01-01

    Cats, as definitive hosts, play an important role in the transmission of Toxoplasma gondii. To determine the seroprevalence and risk factors for T. gondii infection in Dutch domestic cats, serum samples of 450 cats were tested for T. gondii antibodies by indirect ELISA. Binary mixture analysis was

  10. Prevalence of Toxoplasma gondii in Canadian market-age pigs.

    Science.gov (United States)

    Gajadhar, A A; Aramini, J J; Tiffin, G; Bisaillon, J R

    1998-08-01

    During 1991 and 1992, 2,800 market-age pigs were sampled at federally inspected abattoirs from across Canada. Anti-Toxoplasma gondii IgG at titers of > or =1:32 were found in 240 pigs examined by a commercial, latex agglutination test. Seroprevalences ranged from 3.5 to 13.2% in the different regions of the country. Tissue hybridization studies using a previously developed probe demonstrated T. gondii ribosomal RNA in 9 of 36 animals, whereas mouse bioassay testing of heart muscle and diaphragm from all 2,800 pigs failed to demonstrate the presence of infective stages of T. gondii in tissues. Although serology results from this study indicated that Canadian market-age pigs are infected with T. gondii at rates similar to those reported from other parts of North America, mouse bioassay results suggested that Canadian pork products contain low levels of infective organisms. This apparent discrepancy suggests that serological evidence of T. gondii infection in pigs alone does not accurately assess the public health risks associated with consuming improperly cooked pork products.

  11. Meat juice serology for Toxoplasma gondii infection in chickens

    Directory of Open Access Journals (Sweden)

    Alice Vismarra

    2016-01-01

    Full Text Available Toxoplasma gondii is an important foodborne zoonosis. Free-range chickens are at particularly high risk of infection and are also excellent indicators of soil contamination by oocysts. In the present study, hearts of 77 freerange chickens were collected at slaughter. T. gondii meat juice enzyme-linked immunosorbent assay was performed with a commercial kit, following validation with positive controls, from experimentally infected chickens, and negative ones. Out of 77 samples, only 66 gave sufficient meat juice for serology. Of these, 24 (36.4% were positive for T. gondii considering the 5*standard deviation values (calculated on the optical density of negative controls, while all the samples were negative considering sample/positive% values. Parasite-specific polymerase chain reaction was carried out on all samples obtained from heart tissue and none were positive for the presence of T. gondii DNA. Results would suggest that further study on the use of meat juice with a validated serological test to detect T. gondii in chickens could lead to widespread epidemiological studies in this important intermediate host. However, sample collection and test specificity require further evaluation.

  12. Prevalence of encysted Toxoplasma gondii in raptors from Alabama.

    Science.gov (United States)

    Lindsay, D S; Smith, P C; Hoerr, F J; Blagburn, B L

    1993-12-01

    Little is known about the prevalence of encysted Toxoplasma gondii in wild birds. We examined the hearts and breast muscles from 101 raptors for encysted T. gondii. All of the raptors had been submitted for necropsy to the State Veterinary Diagnostic Laboratory, Auburn, Alabama. Tissues were digested in acid-pepsin solution and inoculated into groups of 3-5 laboratory mice. Toxoplasma gondii was isolated from 27 of 101 (26.7%) raptors: 8 of 12 (66.7%) red-shouldered hawks (Buteo lineatus), 13 of 27 (41.1%) red-tailed hawks (Buteo jamaicensis), 1 of 4 (25%) Cooper's hawks (Accipiter cooperi), 1 of 5 (20%) great horned owls (Bubo virginianus), 4 of 15 (26.7%) barred owls (Strix varia), and 1 of 3 (33.3%) kestrels (Falco sparverius). Toxoplasma gondii was not isolated from 3 broad-winged hawks (Buteo platypterus), 3 sharp-shinned hawks (Accipiter striatus), 6 barn owls (Tyto alba), 9 screech owls (Asio otus), a Mississippi kite (Ictinia misisippiensis), 2 golden eagles (Aquila chrysaetos), a bald eagle (Haliaeetus leucocephalus), 4 ospreys (Pandion haliaetus), 4 turkey vultures (Cathartes aura), or 2 black vultures (Coragyps atratus). No significant difference (P > 0.05) in prevalence was detected based on sex using chi-square analysis. Chi-square analysis of the data demonstrated that adult raptors had encysted stages of T. gondii significantly (P < 0.05) more often than did immature raptors.

  13. Toxoplasma gondii coinfection with diseases and parasites in wild rabbits in Scotland.

    Science.gov (United States)

    Mason, Sam; Dubey, J P; Smith, Judith E; Boag, Brian

    2015-09-01

    In wild rabbits (Oryctolagus cuniculus) on an estate in Perthshire, central Scotland, the seroprevalence of Toxoplasma gondii was 18/548 (3·3%). The wild rabbit could be a T. gondii reservoir and it has potential value as a sentinel of T. gondii in environmental substrates. Toxoplasma gondii was associated with female sex (P myxomatosis caused by the virus Myxomatosis cuniculi, the intensity of roundworm eggs, the year or season, rabbit age or distance from farm buildings. Coinfections could have been affected by gestational down regulation of type 1 T helper cells. A sudden influx or release of T. gondii oocysts might have occurred. This is the first report of T. gondii in any wild herbivore in Scotland and also the first report of lapine T. gondii as a coinfection with E. stiedae, M. cuniculi and helminths.

  14. Urine sample used for detection of toxoplasma gondii infection by loop-mediated isothermal amplification (LAMP).

    Science.gov (United States)

    Hu, Xin; Pan, Chang-Wang; Li, Ya-Fei; Wang, Han; Tan, Feng

    2012-02-01

    In this study, a loop-mediated isothermal amplification (LAMP) assay was established to detect Toxoplasma gondii DNA in mice infected with T. gondii PRU strain. This LAMP assay was based on the sequence of highly repetitive B1 gene. The detection limit of T. gondii LAMP assay was 1 pg of T. gondii DNA, which was evaluated using 10-fold serially diluted DNA of cultured parasites. The LAMP assay was also highly specific for T. gondii and able to detect T. gondii DNA in urine of mice treated with dexamethasone at 90 day post infection (p.i.), although this assay could not detect the DNA in mice urine 2-6 days p.i. These results demonstrated that LAMP is effective for evaluation of therapy effectiveness for T. gondii infection. The established LAMP assay may represent a useful and practical tool for the routine diagnosis and therapeutic evaluation of human toxoplasmosis.

  15. Detection of Toxolasma gondii in captive wild felids.

    Science.gov (United States)

    Buddhirongawatr, Ruangrat; Tungsudjai, Siriporn; Chaichoune, Kridsada; Sangloung, Charoonluk; Tantawiwattananon, Nitipan; Phonaknguen, Rassameepen; Sukthana, Yaowalark

    2006-01-01

    Toxoplasma gondii can infect all species of warm-blooded animals, including humans, and causes serious diseases in immunocompromized hosts. Live tachyzoites derived from serial passage in HeLa culture were used in the Sabin-Feldman dye test for detection of Toxoplasma gondii antibody in serum samples of 21 captive wild felids including one fishing cat (Prion nailurus viverrina), one leopard (Panthera pardus), two flat-headed cats (Prion nailurus planiceps), 6 tigers (Panthera tigris), two leopard cats (Felis bengalensis), two clouded leopards (Felis nebulosa), 3 pumas (Puma concolor), and 4 jungle cats (Felis chaus). Antibodies to Toxoplasma gondii were founded in 9 of 21 felids (42.8%). This study revealed that cell culture-derived tachyzoites can be used successfully as a source of live organisms in a gold standard Sabin-Feldman dye test, which is simpler, cheaper and less ethically sensitive than in vivo inoculation.

  16. Lack of cyclophilin B in osteogenesis imperfecta with normal collagen folding.

    Science.gov (United States)

    Barnes, Aileen M; Carter, Erin M; Cabral, Wayne A; Weis, MaryAnn; Chang, Weizhong; Makareeva, Elena; Leikin, Sergey; Rotimi, Charles N; Eyre, David R; Raggio, Cathleen L; Marini, Joan C

    2010-02-11

    Osteogenesis imperfecta is a heritable disorder that causes bone fragility. Mutations in type I collagen result in autosomal dominant osteogenesis imperfecta, whereas mutations in either of two components of the collagen prolyl 3-hydroxylation complex (cartilage-associated protein [CRTAP] and prolyl 3-hydroxylase 1 [P3H1]) cause autosomal recessive osteogenesis imperfecta with rhizomelia (shortening of proximal segments of upper and lower limbs) and delayed collagen folding. We identified two siblings who had recessive osteogenesis imperfecta without rhizomelia. They had a homozygous start-codon mutation in the peptidyl-prolyl isomerase B gene (PPIB), which results in a lack of cyclophilin B (CyPB), the third component of the complex. The proband's collagen had normal collagen folding and normal prolyl 3-hydroxylation, suggesting that CyPB is not the exclusive peptidyl-prolyl cis-trans isomerase that catalyzes the rate-limiting step in collagen folding, as is currently thought. 2010 Massachusetts Medical Society

  17. Cyclosporine A-sensitive, cyclophilin B-dependent endoplasmic reticulum-associated degradation.

    Directory of Open Access Journals (Sweden)

    Riccardo Bernasconi

    2010-09-01

    Full Text Available Peptidyl-prolyl cis/trans isomerases (PPIs catalyze cis/trans isomerization of peptide bonds preceding proline residues. The involvement of PPI family members in protein refolding has been established in test tube experiments. Surprisingly, however, no data is available on the involvement of endoplasmic reticulum (ER-resident members of the PPI family in protein folding, quality control or disposal in the living cell. Here we report that the immunosuppressive drug cyclosporine A (CsA selectively inhibits the degradation of a subset of misfolded proteins generated in the ER. We identify cyclophilin B (CyPB as the ER-resident target of CsA that catalytically enhances disposal from the ER of ERAD-L(S substrates containing cis proline residues. Our manuscript presents the first evidence for enzymatic involvement of a PPI in protein quality control in the ER of living cells.

  18. Cyclophilin B is a functional regulator of hepatitis C virus RNA polymerase.

    Science.gov (United States)

    Watashi, Koichi; Ishii, Naoto; Hijikata, Makoto; Inoue, Daisuke; Murata, Takayuki; Miyanari, Yusuke; Shimotohno, Kunitada

    2005-07-01

    Viruses depend on host-derived factors for their efficient genome replication. Here, we demonstrate that a cellular peptidyl-prolyl cis-trans isomerase (PPIase), cyclophilin B (CyPB), is critical for the efficient replication of the hepatitis C virus (HCV) genome. CyPB interacted with the HCV RNA polymerase NS5B to directly stimulate its RNA binding activity. Both the RNA interference (RNAi)-mediated reduction of endogenous CyPB expression and the induced loss of NS5B binding to CyPB decreased the levels of HCV replication. Thus, CyPB functions as a stimulatory regulator of NS5B in HCV replication machinery. This regulation mechanism for viral replication identifies CyPB as a target for antiviral therapeutic strategies.

  19. Cyclosporine A-Sensitive, Cyclophilin B-Dependent Endoplasmic Reticulum-Associated Degradation

    Science.gov (United States)

    Luban, Jeremy; Molinari, Maurizio

    2010-01-01

    Peptidyl-prolyl cis/trans isomerases (PPIs) catalyze cis/trans isomerization of peptide bonds preceding proline residues. The involvement of PPI family members in protein refolding has been established in test tube experiments. Surprisingly, however, no data is available on the involvement of endoplasmic reticulum (ER)-resident members of the PPI family in protein folding, quality control or disposal in the living cell. Here we report that the immunosuppressive drug cyclosporine A (CsA) selectively inhibits the degradation of a subset of misfolded proteins generated in the ER. We identify cyclophilin B (CyPB) as the ER-resident target of CsA that catalytically enhances disposal from the ER of ERAD-LS substrates containing cis proline residues. Our manuscript presents the first evidence for enzymatic involvement of a PPI in protein quality control in the ER of living cells. PMID:20927389

  20. The clinical implication of serum cyclophilin A in patients with chronic obstructive pulmonary disease

    Directory of Open Access Journals (Sweden)

    Zhang M

    2018-01-01

    Full Text Available Ming Zhang,1 Jingjing Tang,1 Jiafeng Yin,2 Xiaoying Wang,3 Xiangli Feng,1 Xia Yang,1 Hu Shan,1 Qiuhong Zhang,1 Jie Zhang,1 Yali Li1 1Department of Respiratory and Critical Care Medicine, The Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an, 2Department of Laboratory Examination, The Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an, 3Health Examination Center, The Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an, People’s Republic of China Background: Cyclophilin A (CyPA is a secreted molecule that is regulated by inflammatory stimuli. Although inflammation has an important role in the pathogenesis of chronic obstructive pulmonary disease (COPD, little is known regarding the relationship between serum CyPA and COPD. Methods: Ninety-three COPD patients with acute exacerbation were enrolled in the study and were reassessed during the convalescence phase. Eighty-eight controls were matched for age, gender, body mass index, smoking index and comorbidity. The basic clinical information and pulmonary function of all participants were collected. Serum levels of CyPA and other inflammation indexes were further measured. Results: Serum CyPA was significantly increased in convalescent COPD patients compared to healthy controls, and further elevated in COPD patients with acute exacerbation. Serum CyPA positively correlated with serum interleukin-6, matrix metalloproteinase-9 and high-sensitivity C-reactive protein in both the exacerbation and convalescence phases of COPD. Furthermore, it negatively correlated with percent value of forced expiratory volume in 1 second (FEV1% predicted and FEV1/forced vital capacity in convalescent COPD patients. Conclusion: These results suggest that serum CyPA can be used as a potential inflammatory biomarker for COPD and assessment of serum CyPA may reflect the severity of inflammation in COPD. Keywords: cyclophilin A, chronic obstructive pulmonary disease

  1. [Cloning and expressing of cyclophilin B gene from Schistosoma japonnicum and the analysis of immunoprotective effect].

    Science.gov (United States)

    Peng, Jinbiao; Han, Hongxiao; Hong, Yang; Wang, Yan; Guo, Fanji; Shi, Yaojun; Fu, Zhiqiang; Liu, Jinming; Cheng, Guofeng; Lin, Jiaojiao

    2010-03-01

    The present study was intend to clone and express the cDNA encoding Cyclophilin B (CyPB) of Schistosoma japonicum, its preliminary biological function and further immunoprotective effect against schistosome infection in mice. RT-PCR technique was applied to amplify a full-length cDNA encoding protein Cyclophilin B (Sj CyPB) from schistosomula cDNA. The expression profiles of Sj CyPB were determined by Real-time PCR using the template cDNAs isolated from 7, 13, 18, 23, 32 and 42 days parasites. The cDNA containing the Open Reading Frame of CyPB was then subcloned into a pGEX-6P-1 vector and transformed into competent Escherichia coli BL21 for expressing. The recombinant protein was renaturated, purified and its antigenicity were detected by Western blotting, and the immunoprotective effect induced by recombinant Sj CyPB was evaluated in Balb/C mice. The cDNA containing the ORF of Sj CyPB was cloned with the length of 672 base pairs, encoding 223 amino acids. Real-time PCR analysis revealed that the gene had the highest expression in 18-day schistosomula, suggesting that Sj CyPB was schistosomula differentially expressed gene. The recombinant protein showed a good antigenicity detected by Western blotting. Animal experiment indicated that the vaccination of recombinant CyPB protein in mice led to 31.5% worm and 41.01% liver egg burden reduction, respectively, compared with those of the control. A full-length cDNA differentially expressed in schistosomula was obtained. The recombinant Sj CyPB protein could induce partial protection against schistosome infection.

  2. Toxoplasma gondii infections in sheep in Sicily, southern Italy

    DEFF Research Database (Denmark)

    Vesco, G; Buffolano, W; La Chiusa, S

    2007-01-01

    is very prevalent, and eating unprocessed sheep and lamb meat has a high risk of transmitting infections to humans. The presence of cats on the farm, farm size and using surface water as drinking water for the animals were risk factors for infection in sheep, with age as a significant confounder...... outdoor on pasture and only one was claiming organic farming. Having cats on the farm, age of the animals, farm size and the use of surface water sources for drinking were all significantly associated with T. gondii-infected animals on the farm. T. gondii infection in mutton used for human consumption...

  3. A dual inhibitor against prolyl isomerase Pin1 and cyclophilin discovered by a novel real-time fluorescence detection method

    International Nuclear Information System (INIS)

    Mori, Tadashi; Hidaka, Masafumi; Lin, Yi-Chin; Yoshizawa, Ibuki; Okabe, Takayoshi; Egashira, Shinichiro; Kojima, Hirotatsu; Nagano, Tetsuo; Koketsu, Mamoru; Takamiya, Mari; Uchida, Takafumi

    2011-01-01

    Research highlights: → A Pin1 (prolyl isomerase) inhibitor, TME-001, has been discovered by using a new established high-throughput screening method. → The TME-001 showed a cell-active inhibition with lower cytotoxic effect than known Pin1 inhibitors. → Kinetic analyses revealed that the TME-001 is the first compound that exhibits dual inhibition of Pin1 and another type of prolyl isomerase, cyclophilin. → Thus, similarities of structure and reaction mechanism between Pin1 and cyclophilin are proposed. -- Abstract: Pin1, a peptidyl prolyl cis/trans isomerase (PPIase), is a potential target molecule for cancer, infectious disease, and Alzheimer's disease. We established a high-throughput screening method for Pin1 inhibitors, which employs a real-time fluorescence detector. This screening method identified 66 compounds that inhibit Pin1 out of 9756 compounds from structurally diverse chemical libraries. Further evaluations of surface plasmon resonance methods and a cell proliferation assay were performed. We discovered a cell-active inhibitor, TME-001 (2-(3-chloro-4-fluoro-phenyl)-isothiazol-3-one). Surprisingly, kinetic analyses revealed that TME-001 is the first compound that exhibits dual inhibition of Pin1 (IC 50 = 6.1 μM) and cyclophilin, another type of PPIase, (IC 50 = 13.7 μM). This compound does not inhibit FKBP. This finding suggests the existence of similarities of structure and reaction mechanism between Pin1 and cyclophilin, and may lead to a more complete understanding of the active sites of PPIases.

  4. Biochemical Characterization of the Prolyl 3-Hydroxylase 1·Cartilage-associated Protein·Cyclophilin B Complex*

    Science.gov (United States)

    Ishikawa, Yoshihiro; Wirz, Jackie; Vranka, Janice A.; Nagata, Kazuhiro; Bächinger, Hans Peter

    2009-01-01

    The rough endoplasmic reticulum-resident protein complex consisting of prolyl 3-hydroxylase 1 (P3H1), cartilage-associated protein (CRTAP), and cyclophilin B (CypB) can be isolated from chick embryos on a gelatin-Sepharose column, indicating some involvement in the biosynthesis of procollagens. Prolyl 3-hydroxylase 1 modifies a single proline residue in the α chains of type I, II, and III collagens to (3S)-hydroxyproline. The peptidyl-prolyl cis-trans isomerase activity of cyclophilin B was shown previously to catalyze the rate of triple helix formation. Here we show that cyclophilin B in the complex shows peptidyl-prolyl cis-trans isomerase activity and that the P3H1·CRTAP·CypB complex has another important function: it acts as a chaperone molecule when tested with two classical chaperone assays. The P3H1·CRTAP·CypB complex inhibited the thermal aggregation of citrate synthase and was active in the denatured rhodanese refolding and aggregation assay. The chaperone activity of the complex was higher than that of protein-disulfide isomerase, a well characterized chaperone. The P3H1·CRTAP·CypB complex also delayed the in vitro fibril formation of type I collagen, indicating that this complex is also able to interact with triple helical collagen and acts as a collagen chaperone. PMID:19419969

  5. Evaluation of Porin Interaction with Adenine Nucleotide Translocase and Cyclophilin-D Proteins after Brain Ischemia and Reperfusion

    Directory of Open Access Journals (Sweden)

    Mohammad Ali Atlasi

    2011-01-01

    Full Text Available Objective (s Porin is a mitochondrial outer membrane channel, which usually functions as the pathway for the movement of various substances in and out of the mitochondria and is considered to be a component of the permeability transition (PT pore complex that plays a role in the PT. We addressed the hypothesis that porin interacts with other mitochondrial proteins after ischemic injury.Materials and MethodsFor this purpose, we used in vivo 4-vessel occlusion model of rat brain and porin purification method by hydroxyapatite column. After SDS gel electrophoresis and silver nitrate staining, Western blotting was done for porin, adenine nucleotide translocase and cyclophilin-D proteins.Results Porin was purified from mitochondrial mixture in ischemic brain and control groups. Investigation of interaction of adenine nucleotide transposes (ANT and cyclophilin-D with porin by Western blotting showed no proteins co-purified with porin from injured tissues.Conclusion The present study implies that there may not be interaction between porin, and ANT or cyclophilin-D, and if there is any, it is not maintained during the purification procedure.

  6. SPORULATION AND SURVIVAL OF TOXOPLASMA GONDII OOCYSTS IN SEA WATER

    Science.gov (United States)

    Since 1992, we have been collaborating in studies on southern sea otters (Enhdyra lutris nereis) as part of a program to define factors which may be responsible for limiting the growth of the southern sea otter population. We previously demonstrated Toxoplasma gondii in sea otter...

  7. Detection of Toxoplasma gondii DNA in Brazilian oysters (Crassostrea rhizophorae).

    Science.gov (United States)

    Ribeiro, L A; Santos, L K N S S; Brito, P A; Maciel, B M; Da Silva, A V; Albuquerque, G R

    2015-05-04

    The aim of this study was to detect evidence of Toxoplasma gondii using polymerase chain reaction (PCR)-based techniques in oysters (Crassostrea rhizophorae) obtained from the southern coastal region of Bahia, Brazil. A total of 624 oysters were collected, and the gills and digestive glands were dissected. Each tissue sample was separated into pools containing tissues (of the same type) from three animals, leading to a total of 416 experimental samples for analysis (208 samples each from the gills and digestive glands). Molecular analysis using PCR-based detection of the T. gondii AF 146527 repetitive fragment yielded negative results for all samples. However, when nested-PCR was used for detection of the T. gondii SAG-1 gene, 17 samples were positive, with the gills being the tissue with maximal detection of the parasite. These positive results were confirmed by sample sequencing. It is therefore suggested that C. rhizophorae oysters are capable of filtering and retaining T. gondii oocysts in their tissue. This represents a risk to public health because they are traditionally ingested in natura.

  8. Anti- toxoplasma gondii activity of constituents from Balsamocitrus ...

    African Journals Online (AJOL)

    Isolation, characterization and anti-Toxoplasma gondii activity of constituents from the CH2Cl2/MeOH (1/1) extract of the roots of the cameroonian plant Balsamocitrus camerunensis L. were investigated in this study. Four known coumarins derivatives were isolated, namely, marmin (1), imperatorin (2), xanthoxyletin (3), ...

  9. Antibodies to Toxoplasma gondii in Backyard and Roaming Pigs ...

    African Journals Online (AJOL)

    Toxoplasma gondii, the etiologic agent of Toxoplasmosis, can be transmitted to pigs through the ingestion of oocysts, and to humans through consumption of pork containing viable cysts causing neonatal deaths and abortion in animals, and opportunistic infections in immunocompromised humans. The objective of this ...

  10. Cloning and expression of Toxoplasma gondii tachyzoite P22 protein

    African Journals Online (AJOL)

    Delay in diagnosis of Toxoplasma gondii infection in pregnant women who have been infected during the first trimester of gestation can lead to death of her fetus. Serological tests based on recombinant proteins are the main diagnosis methods for the detection of anti Toxoplasma antibody in serum samples. The aim of this ...

  11. Isolation of Toxoplasma gondii from horse meat in Egypt.

    Science.gov (United States)

    Shaapan, R M; Ghazy, A A

    2007-01-01

    Portions of heart, liver, skeletal and diaphragmatic muscles obtained from 150 slaughtered horses at Giza-Zoo abattoir were used for bioassays in mice and cats. T. gondii tachyzoites were isolated successfully from the peritoneal exudates of the inoculated mice 6-8 days post inoculation with pooled horse tissues. Whereas, T. gondii tissue cysts containing bradyzoites were detected in the impression smears of mice brain on the 45th days or more post infection. The oocysts were detected in feces of cats 3-6 days post feeding on horse tissues containing tissue cysts. The oocysts became sporulated within 3-5 days in 2.5% Potassium dichromate. A total of 79 out of 150 horse meat samples were found to be infected with an incidence rate of 52.6 %. This is the first trial for isolation of T. gondii infective stages from horses in Egypt. Moreover, this study pointed out to the high infection rate of T. gondii in horse meat which may be considered as an important source of infection to wild zoo-animals in Egypt and humans in some countries if consumed raw or insufficiently cooked.

  12. Seroprevalence of Toxoplasma gondii infection in feral cats in Qatar.

    Science.gov (United States)

    Boughattas, Sonia; Behnke, Jerzy; Sharma, Aarti; Abu-Madi, Marawan

    2017-01-18

    Cats are essential in the life cycle of Toxoplasma gondii as they can shed the environmentally resistant oocysts after acquiring infection. Human populations living in cities with high densities of feral cats are therefore likely to be at risk of infection. The current study is the first to estimate the seroprevalence of T. gondii in the feral cat population in Qatar. We investigated the seroprevalence of T. gondii among 495 adult cats from urban and suburban districts in Qatar. Using results from the Modified Agglutination Test, we fitted statistical models with host sex, area and season as explanatory factors and seropositivity as the outcome. The analysis revealed an overall seroprevalence of 82%. Seroprevalence was significantly higher in the summer season (P = 0.006). No significant difference was detected (P > 0.05) between seroprevalence in female and male cats and in cats from urban and suburban districts of Qatar. Despite the seasonal difference, the observed seroprevalence of T. gondii suggests high environmental contamination throughout the year, with some female cats generating more intense responses compared to males. Both findings merit further investigations.

  13. Structure of Toxoplasma gondii fructose-1,6-bisphosphate aldolase

    International Nuclear Information System (INIS)

    Boucher, Lauren E.; Bosch, Jürgen

    2014-01-01

    The structure of T. gondii fructose-1,6-bisphosphate aldolase, a glycolytic enzyme and structural component of the invasion machinery, was determined to a resolution of 2.0 Å. The apicomplexan parasite Toxoplasma gondii must invade host cells to continue its lifecycle. It invades different cell types using an actomyosin motor that is connected to extracellular adhesins via the bridging protein fructose-1,6-@@bisphosphate aldolase. During invasion, aldolase serves in the role of a structural bridging protein, as opposed to its normal enzymatic role in the glycolysis pathway. Crystal structures of the homologous Plasmodium falciparum fructose-1,6-bisphosphate aldolase have been described previously. Here, T. gondii fructose-1,6-bisphosphate aldolase has been crystallized in space group P22 1 2 1 , with the biologically relevant tetramer in the asymmetric unit, and the structure has been determined via molecular replacement to a resolution of 2.0 Å. An analysis of the quality of the model and of the differences between the four chains in the asymmetric unit and a comparison between the T. gondii and P. falciparum aldolase structures is presented

  14. Toxoplasma gondii decreases the reproductive fitness in mice

    Czech Academy of Sciences Publication Activity Database

    Dvořáková-Hortová, K.; Šídlová, A.; Děd, Lukáš; Hladovcová, D.; Vieweg, M.; Weidner, W.; Steger, K.; Stopka, P.; Paradowska-Dogan, A.

    2014-01-01

    Roč. 9, č. 6 (2014), s. 1-11 E-ISSN 1932-6203 R&D Projects: GA MŠk(CZ) ED1.1.00/02.0109 Institutional support: RVO:86652036 Keywords : Toxoplasma gondii * reproductive fitness * DNA methylation Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.234, year: 2014

  15. Seroprevalence and risk factors of Toxoplasma gondii infection ...

    African Journals Online (AJOL)

    The infection is also associated with human immune deficiency virus/acquired immunodeficiency syndrome (HIV/AIDS). In Rwanda, the burden and risk factors of T. gondii infection among pregnant women and among HIV infected pregnant women is largely unknown. This cross-sectional study aimed at determining the ...

  16. Sero-prevalence of Toxoplasma gondii in Danish pigs

    DEFF Research Database (Denmark)

    Kofoed, Kristina Grønbech; Vorslund-Kiær, Mia; Nielsen, Henrik Vedel

    2017-01-01

    In 2015, the World Health Organisation rated toxoplasmosis as one of the most important food borne zoonotic diseases in the world. In addition, recent studies have associated Toxoplasma gondii sero-positivity with severe mental illnesses such as schizophrenia. Intake of raw or insufficiently cooked...

  17. The neurotropic parasite Toxoplasma gondii increases dopamine metabolism

    Science.gov (United States)

    The common parasite Toxoplasma gondii induces behavioral alterations in its hosts including phenotypes increasing the likelihood of its transmission in rodents and reports of psychobehavioral alterations in humans. We have found that elevated levels of dopamine are associated with the encysted stage...

  18. Hepatoprotective activity of Thymus vulgaris extract against Toxoplasma gondii infection

    Directory of Open Access Journals (Sweden)

    Nagwa Mostafa El-Sayed

    2017-05-01

    Full Text Available Objective: To evaluate the hepatoprotective activity of Thymus vulgaris (T. vulgaris extract against Toxoplasma gondii (T. gondii infection in experimentally infected mice. Methods: Sixty mice were divided into six groups (Group I–Group VI. Group I was normal control (non-infected, non-treated; Group II was non-infected and treated with T. vulgaris extract (500 mg/kg; Group III was T. gondii infected-non-immunosuppressed control; Group IV consisted of infected immunosuppressed mice; Group V was infected and treated with T. vulgaris extract; Group VI consisted of infected immunosuppressed mice treated with T. vulgaris extract. Hepatoprotective effect of T. vulgaris extract was evaluated by histopathological examination of tissue sections stained with hematoxylin and eosin, determination of liver function parameters (alanine aminotransaminase, aspartate aminotransaminase and alkaline phosphates, total bilirubin, total protein concentrations and assessment of hepatocytes genotoxicity by comet assay.Antigenotoxic effect of T. vulgaris was assessed by several comet assay parameters that were provided by the image analysis software, including % tailed cells, % of DNA in the tail, tail length, and tail moment. Results: Treatment with T. vulgaris in both Groups V and VI improved T. gondii induced pathological lesions in the infected liver that regressed to near the normal picture especially in Group V. Also, it restored the altered values of liver function parameters near to the normal levels significantly (P < 0.05 compared with Groups III and IV respectively. Regarding comet assay parameters, all of them were significantly increased (P < 0.05 after T. gondii infection (Group III and reached the greatest values in infected immunosuppressed group (Group IV compared to the normal controls (Group I. With treatment by T. vulgaris in Groups V and VI, there was a significant decrease (P < 0.05 in all values compared to Groups III and V respectively. The

  19. Toxoplasma gondii abortion storm in sheep on a Texas farm and isolation of mouse virulent atypical genotype T. gondii from an aborted lamb from a chronically infected ewe

    Science.gov (United States)

    Sheep are commonly infected with the protozoan parasite, Toxoplasma gondii. Infection may cause early embryonic death and resorption, fetal death and mummification, abortion, stillbirth, and neonatal death. Most sheep acquire T. gondii infection after birth. Recent studies reported that repeat ovine...

  20. Seroprevalensi Toxoplasma gondii pada Kambing dan Bioassay Patogenitasnya pada Kucing

    Directory of Open Access Journals (Sweden)

    Ni Made Yunik Novita Dewi Dewi

    2013-11-01

    Full Text Available Normal 0 false false false EN-US X-NONE X-NONE The study aimed to determine seroprevalence of Toxoplasmosis in goats sloughtered at Kampung Jawa, Denpasar, Bali and to evaluate their pathogenicities through bioassay in cats.One hundred serums and meats of goats were collected. Anti-Toxoplasma gondii antibody was determined using Indirect Haemaglutination (IHA test. The pathogenicity bioassay of Toxoplasma gondii was carried out through inoculating the meats of goats which had seropositive of Toxoplasma gondii to the cats. The pathogenicity was evaluated using the intensity of oocyte sheding from the cats. The result showed that the seroprevalence of Toxoplasmosis was 46%. There was not significant difference between pathogenicity of Toxoplasma gondii in cat inoculated with meat of goat which had a high and low titer of antibody against Toxoplasma gondii. /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; text-align:justify; line-height:150%; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin;}

  1. Neospora caninum and Toxoplasma gondii antibody prevalence in Alaska wildlife.

    Science.gov (United States)

    Stieve, Erica; Beckmen, Kimberlee; Kania, Stephen A; Widner, Amanda; Patton, Sharon

    2010-04-01

    Free-ranging caribou and moose populations in some regions of Alaska undergo periodic declines in numbers. Caribou and moose are managed by the state as valuable resources for not only sustenance and subsistence, but also for cultural heritage. Incidence and prevalence of diseases that may impact herd health and recruitment from year to year are relevant to management decisions aimed to protect the long-term viability of these herds. Neospora caninum and Toxoplasma gondii are two apicomplexan parasites that can cause neurologic disease and abortions in their intermediate hosts and less frequently cause disease in their definitive hosts. The definitive hosts of N. caninum and T. gondii are canids and felids, respectively, and prevalence in the environment is in part dependent on maintenance of the life cycle through the definitive hosts. Serum samples from caribou (Rangifer tarandus, n=453), wolf (Canis lupus, n=324), moose (Alces alces, n=201), black-tailed deer (Odocoileus hemionus, n=55), coyote (Canis latrans, n=12), and fox (Vulpes vulpes, n=9) collected in Alaska were assayed for N. caninum- and T. gondii-reactive antibodies with an immunofluorescent antibody test (IFAT) and a modified agglutination test (MAT), respectively. Seroprevalence of N. caninum was greater in caribou (11.5%) than in wolves (9.0%), moose (0.5%), or black-tailed deer (0%). Seroprevalence of T. gondii was greater in wolves (17.8%) than in caribou (0.4%), moose (0%), or black-tailed deer (0%). Seroprevalence of N. caninum and T. gondii were 16.7% and 0.0% in coyotes and 0.0% and 12.5% in fox, but small sample sizes prevented further analysis. Antibodies to N. caninum in young caribou compared to adult caribou suggest that vertical transmission may be an important component of new infections in Alaskan caribou. The spatial distribution of antibody-positive individuals across Alaska may reflect differences in frequency of definitive hosts and alteration of predation patterns among regions.

  2. Epidemiology and pathology of Toxoplasma gondii in free-ranging California sea lions (Zalophus californianus)

    OpenAIRE

    Carlson-Bremer, D; Colegrove, KM; Gulland, FMD; Conrad, PA; Mazet, JAK; Johnson, CK

    2015-01-01

    © Wildlife Disease Association 2015. The coccidian parasite Toxoplasma gondii infects humans and warm-blooded animals worldwide. The ecology of this parasite in marine systems is poorly understood, although many marine mammals are infected and susceptible to clinical toxoplasmosis. We summarized the lesions associated with T. gondii infection in the California sea lion (Zalophus californianus) population and investigated the prevalence of and risk factors associated with T. gondii exposure, a...

  3. Seroprevalence of Toxoplasma gondii in free-ranging and feral cats on Amami Oshima Island, Japan

    OpenAIRE

    MATSUU, Aya; YOKOTA, Shin-ichi; ITO, Keiko; MASATANI, Tatsunori

    2017-01-01

    On Amami Oshima Island, free-ranging and feral cats are harmful to wildlife populations. In this study, the seroprevalence of Toxoplasma gondii in these cats was examined using a newly developed Gaussia luciferase immunoprecipitation system assay. Of 1,363 cats, 123 cats (9.0%) was positive for T. gondii. The prevalence was significantly different in different areas; among cats in the rural area, where many wild animals live, including endangered species, T. gondii infection was more prevalen...

  4. Seroepidemiology of Toxoplasma gondii infection in pregnant women in a public hospital in northern Mexico

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    Díaz-García Juan

    2006-07-01

    Full Text Available Abstract Background Toxoplasma gondii (T. gondii infection in pregnant women represents a risk for congenital disease. There is scarce information about the epidemiology of T. gondii infection in pregnant women in Mexico. Therefore, we sought to determine the prevalence of T. gondii infection and associated socio-demographic, clinical and behavioural characteristics in a population of pregnant women of Durango City, Mexico. Methods Three hundred and forty three women seeking prenatal care in a public hospital of Durango City in Mexico were examined for T. gondii infection. All women were tested for anti-T. gondii IgM and IgG antibodies by using IMx Toxo IgM and IMx Toxo IgG 2.0 kits (Abbott Laboratories, Abbott Park, IL, USA, respectively. Socio-demographic, clinical and behavioural characteristics from each participant were also obtained. Results Twenty one out of the 343 (6.1% women had IgG anti-T. gondii antibodies. None of the 343 women had IgM anti-T. gondii antibodies. Multivariate analysis using logic regression showed that T. gondii infection was associated with living in a house with soil floor (adjusted OR = 7.16; 95% CI: 1.39–36.84, residing outside of Durango State (adjusted OR = 4.25; 95% CI: 1.72–10.49, and turkey meat consumption (adjusted OR = 3.85; 95% CI: 1.30–11.44. Other characteristics as cat contact, gardening, and food preferences did not show any association with T. gondii infection. Conclusion The prevalence of T. gondii infection in pregnant women of Durango City is low as compared with those reported in other regions of Mexico and the majority of other countries. Poor housing conditions as soil floors, residing in other Mexican States, and turkey meat consumption might contribute to acquire T. gondii infection.

  5. ELISA-seroprevalence of Toxoplasma gondii in draught horses in Greater Cairo, Egypt.

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    Haridy, Fouad M; Shoukry, Nahla M; Hassan, Aly Awad; Morsy, Tosson A

    2009-12-01

    Toxoplasma gondii is one of the important zoonotic parasites of worldwide. In this paper the seroprevalence of T. gondii in draught horses (3-15 years) including 90 males and 10 females in the first half of the year 2009 was studied. The result showed that the overall ELISA-T. gondii antibodies were 25% of the horses in Greater Cairo, 50% (females) and 22.2% (males).

  6. Seropositivity of Toxoplasma gondii in domestic donkeys (Equus asinus) and isolation of T. gondii from farm cats.

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    Dubey, J P; Ness, S L; Kwok, O C H; Choudhary, S; Mittel, L D; Divers, T J

    2014-01-17

    Donkeys (Equus asinus) are used as both companion and working animals throughout the world and in some countries, their meat and milk are used for human consumption. Here we report the first serological survey of Toxoplasma gondii in donkeys in the United States. Serum samples from 373 donkeys from eight farms in five states were tested for T. gondii antibodies by the modified agglutination test (MAT). Twenty-four of 373 (6.4%) of donkeys were seropositive, with MAT titers ranging from 25 to ≥ 200. All seropositive donkeys were Miniature breed. Seropositivity prevalence was 7.0% in female donkeys (20/282) and 4.1% in male donkeys (4/91). No donkeys less than 24 months of age (129) were seropositive, suggesting postnatal transmission of infection. Domestic cats were present on six of the eight farms. Three cats from one farm had MAT titers of 200. Viable T. gondii was isolated from the hearts of two cats, but not from brain tissues. Genotyping of isolate DNA extracted from culture-derived tachyzoites using 10 PCR-restriction fragment length polymorphism (RFLP) markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, PK1, L358 and Apico loci) revealed that both isolates were clonal Type II (ToxoDB PCR-RFLP genotype #1). This is the first serological survey for T. gondii in donkeys in the United States, and suggests that donkey milk and meat should be considered as a potential source for human infection. The role of barn cats in the transmission of T. gondii to donkeys on farms warrents further investigation. Copyright © 2013 Elsevier B.V. All rights reserved.

  7. Detection of infection with toxoplasma Gondii in manatees (Trichechus inunguis) of the peruvian amazon

    International Nuclear Information System (INIS)

    Mathews Delgado, Patrick; Sanchez Perea, Nofre; Mathews Delgado, John Paul; Biffi Garcia, Claudia; Malheiros, Antonio Francisco; Garcia Davila, Carmen Rosa

    2013-01-01

    The Amazonian manatee (trichechus inunguis) is an aquatic mammal that inhabits freshwater environments and is endemic to the amazon basin. The presence of Toxoplasma gondii antibodies was investigated in 19 manatees in one rescue unit in the northern region of Peru. Antibodies to T. gondii were detected in 12 (63.2 %) of 19 animals by using the modified agglutination test (titer, 1:25), and no association between sex and age of the animals and the presence of T. gondii antibodies was observed (p < 0.05). the results suggest a contamination by T. gondii oocysts in the aquatic environment where these animals live.

  8. A novel multifunctional oligonucleotide microarray for Toxoplasma gondii

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    Chen Feng

    2010-10-01

    Full Text Available Abstract Background Microarrays are invaluable tools for genome interrogation, SNP detection, and expression analysis, among other applications. Such broad capabilities would be of value to many pathogen research communities, although the development and use of genome-scale microarrays is often a costly undertaking. Therefore, effective methods for reducing unnecessary probes while maintaining or expanding functionality would be relevant to many investigators. Results Taking advantage of available genome sequences and annotation for Toxoplasma gondii (a pathogenic parasite responsible for illness in immunocompromised individuals and Plasmodium falciparum (a related parasite responsible for severe human malaria, we designed a single oligonucleotide microarray capable of supporting a wide range of applications at relatively low cost, including genome-wide expression profiling for Toxoplasma, and single-nucleotide polymorphism (SNP-based genotyping of both T. gondii and P. falciparum. Expression profiling of the three clonotypic lineages dominating T. gondii populations in North America and Europe provides a first comprehensive view of the parasite transcriptome, revealing that ~49% of all annotated genes are expressed in parasite tachyzoites (the acutely lytic stage responsible for pathogenesis and 26% of genes are differentially expressed among strains. A novel design utilizing few probes provided high confidence genotyping, used here to resolve recombination points in the clonal progeny of sexual crosses. Recent sequencing of additional T. gondii isolates identifies >620 K new SNPs, including ~11 K that intersect with expression profiling probes, yielding additional markers for genotyping studies, and further validating the utility of a combined expression profiling/genotyping array design. Additional applications facilitating SNP and transcript discovery, alternative statistical methods for quantifying gene expression, etc. are also pursued at

  9. The Activation of Phytophthora Effector Avr3b by Plant Cyclophilin is Required for the Nudix Hydrolase Activity of Avr3b.

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    Kong, Guanghui; Zhao, Yao; Jing, Maofeng; Huang, Jie; Yang, Jin; Xia, Yeqiang; Kong, Liang; Ye, Wenwu; Xiong, Qin; Qiao, Yongli; Dong, Suomeng; Ma, Wenbo; Wang, Yuanchao

    2015-08-01

    Plant pathogens secrete an arsenal of effector proteins to impair host immunity. Some effectors possess enzymatic activities that can modify their host targets. Previously, we demonstrated that a Phytophthora sojae RXLR effector Avr3b acts as a Nudix hydrolase when expressed in planta; and this enzymatic activity is required for full virulence of P. sojae strain P6497 in soybean (Glycine max). Interestingly, recombinant Avr3b produced by E. coli does not have the hydrolase activity unless it was incubated with plant protein extracts. Here, we report the activation of Avr3b by a prolyl-peptidyl isomerase (PPIase), cyclophilin, in plant cells. Avr3b directly interacts with soybean cyclophilin GmCYP1, which activates the hydrolase activity of Avr3b in a PPIase activity-dependent manner. Avr3b contains a putative Glycine-Proline (GP) motif; which is known to confer cyclophilin-binding in other protein substrates. Substitution of the Proline (P132) in the putative GP motif impaired the interaction of Avr3b with GmCYP1; as a result, the mutant Avr3bP132A can no longer be activated by GmCYP1, and is also unable to promote Phytophthora infection. Avr3b elicits hypersensitive response (HR) in soybean cultivars producing the resistance protein Rps3b, but Avr3bP132A lost its ability to trigger HR. Furthermore, silencing of GmCYP1 rendered reduced cell death triggered by Avr3b, suggesting that GmCYP1-mediated Avr3b maturation is also required for Rps3b recognition. Finally, cyclophilins of Nicotiana benthamiana can also interact with Avr3b and activate its enzymatic activity. Overall, our results demonstrate that cyclophilin is a "helper" that activates the enzymatic activity of Avr3b after it is delivered into plant cells; as such, cyclophilin is required for the avirulence and virulence functions of Avr3b.

  10. Mitochondrial permeability transition and cell death: the role of cyclophilin D

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    Sabzali eJavadov

    2013-04-01

    Full Text Available Mitochondria serve as a powerhouse which provides near 90% of ATP necessary for cell life. However, recent studies provide strong evidence that mitochondria also play a central role in cell death. Irreversible mitochondrial permeability transition (mPT at high conductance in response to oxidative or other cellular stresses is accompanied by formation of pathological and non-specific mPT pores (mPTP in the inner membrane of mitochondria. Mitochondrial PTP can serve as a target to prevent cell death under pathological conditions such as cardiac and brain ischemia/reperfusion injury and diabetes. On the other hand, mPTP can be used as an executioner to specifically induce cell death thus blocking tumorigenesis in cancer diseases. Despite many studies, the molecular identity of the mPTP remains unclear. At present, cyclophilin D (CyP-D represents the only mPTP protein which plays an essential role in pore formation. This review will discuss direct and indirect mechanisms underlying CyP-D interaction with a target protein of the mPTP complex. Understanding of the mechanisms of mPTP formation will be helpful to further develop new pharmacological agents targeting mitochondria-mediated cell death.

  11. Regulatory polymorphisms in the cyclophilin A gene, PPIA, accelerate progression to AIDS.

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    Ping An

    2007-06-01

    Full Text Available Human cyclophilin A, or CypA, encoded by the gene peptidyl prolyl isomerase A (PPIA, is incorporated into the HIV type 1 (HIV-1 virion and promotes HIV-1 infectivity by facilitating virus uncoating. We examined the effect of single nucleotide polymorphisms (SNPs and haplotypes within the PPIA gene on HIV-1 infection and disease progression in five HIV-1 longitudinal history cohorts. Kaplan-Meier survival statistics and Cox proportional hazards model were used to assess time to AIDS outcomes. Among eight SNPs tested, two promoter SNPs (SNP3 and SNP4 in perfect linkage disequilibrium were associated with more rapid CD4(+ T-cell loss (relative hazard = 3.7, p = 0.003 in African Americans. Among European Americans, these alleles were also associated with a significant trend to more rapid progression to AIDS in a multi-point categorical analysis (p = 0.005. Both SNPs showed differential nuclear protein-binding efficiencies in a gel shift assay. In addition, one SNP (SNP5 located in the 5' UTR previously shown to be associated with higher ex vivo HIV-1 replication was found to be more frequent in HIV-1-positive individuals than in those highly exposed uninfected individuals. These results implicate regulatory PPIA polymorphisms as a component of genetic susceptibility to HIV-1 infection or disease progression, affirming the important role of PPIA in HIV-1 pathogenesis.

  12. Ricinus communis cyclophilin: functional characterisation of a sieve tube protein involved in protein folding.

    Science.gov (United States)

    Gottschalk, Maren; Dolgener, Elmar; Xoconostle-Cázares, Beatriz; Lucas, William J; Komor, Ewald; Schobert, Christian

    2008-09-01

    The phloem translocation stream of the angiosperms contains a special population of proteins and RNA molecules which appear to be produced in the companion cells prior to being transported into the sieve tube system through the interconnecting plasmodesmata. During this process, these non-cell-autonomous proteins are thought to undergo partial unfolding. Recent mass spectroscopy studies identified peptidyl-prolyl cis-trans isomerase (PPIases) as potential molecular chaperones functioning in the phloem translocation stream (Giavalisco et al. 2006). In the present study, we describe the cloning and characterisation of a castor bean phloem cyclophilin, RcCYP1 that has high peptidyl-prolyl cis-trans isomerase activity. Equivalent enzymatic activity was detected with phloem sap or purified recombinant (His)(6)-tagged RcCYP1. Mass spectrometry analysis of proteolytic peptides, derived from a 22 kDa band in HPLC-fractionated phloem sap, immunolocalisation studies and Western analysis of proteins extracted from castor bean tissues/organs indicated that RcCYP1 is an abundant protein in the companion cell-sieve element complex. Microinjection experiments established that purified recombinant (His)(6)-RcCYP1 can interact with plasmodesmata to both induce an increase in size exclusion limit and mediate its own cell-to-cell trafficking. Collectively, these findings support the hypothesis that RcCYP1 plays a role in the refolding of non-cell-autonomous proteins after their entry into the phloem translocation stream.

  13. Structural and functional characterization of the interaction between cyclophilin B and a heparin-derived oligosaccharide.

    Science.gov (United States)

    Hanoulle, Xavier; Melchior, Aurélie; Sibille, Nathalie; Parent, Benjamin; Denys, Agnès; Wieruszeski, Jean-Michel; Horvath, Dragos; Allain, Fabrice; Lippens, Guy; Landrieu, Isabelle

    2007-11-23

    The chemotaxis and integrin-mediated adhesion of T lymphocytes triggered by secreted cyclophilin B (CypB) depend on interactions with both cell surface heparan sulfate proteoglycans (HSPG) and the extracellular domain of the CD147 membrane receptor. Here, we use NMR spectroscopy to characterize the interaction of CypB with heparin-derived oligosaccharides. Chemical shift perturbation experiments allowed the precise definition of the heparan sulfate (HS) binding site of CypB. The N-terminal extremity of CypB, which contains a consensus sequence for heparin-binding proteins was modeled on the basis of our experimental NMR data. Because the HS binding site extends toward the CypB catalytic pocket, we measured its peptidyl-prolyl cis-trans isomerase (PPIase) activity in the absence or presence of a HS oligosaccharide toward a CD147-derived peptide. We report the first direct evidence that CypB is enzymatically active on CD147, as it is able to accelerate the cis/trans isomerization of the Asp(179)-Pro(180) bond in a CD147-derived peptide. However, HS binding has no significant influence on this PPIase activity. We thus conclude that the glycanic moiety of HSPG serves as anchor for CypB at the cell surface, and that the signal could be transduced by CypB via its PPIase activity toward CD147.

  14. Phase 1 clinical study of cyclophilin B peptide vaccine for patients with lung cancer.

    Science.gov (United States)

    Gohara, Rumi; Imai, Nobue; Rikimaru, Toru; Yamada, Akira; Hida, Naoya; Ichiki, Masao; Kawamoto, Mayumi; Matsunaga, Kazuko; Ashihara, Junko; Yano, Sayoko; Tamura, Mayumi; Ohkouchi, Shinya; Yamana, Hideaki; Oizumi, Kotaro; Itoh, Kyogo

    2002-01-01

    Cyclophilin B (CypB) possesses two antigenic epitopes (CypB(84-92) and CypB(91-99) ) recognized by HLA-A24-restricted and tumor-specific cytotoxic T lymphocytes (CTLs). To determine the safety of CypB-derived peptides and its ability to generate antitumor immune responses, patients with advanced lung cancer received subcutaneous vaccinations of these peptides or their modified peptides. All 16 patients were vaccinated with CypB(91-99) or its modified peptide, whereas only two patients were vaccinated with the modified CypB(84-92), as immediate-type hypersensitivity to CypB(84-92) or its modified peptide was observed in the remaining patients. No severe adverse events were associated with the vaccination. No significant increase in cellular responses to either peptides or tumor cells was observed in the postvaccination PBMCs by the conventional CTL assays in any patients tested. These results suggest that the vaccination of CypB(91-99) peptide was safe, but failed to induce objective immune responses at this regimen.

  15. Hypoxia induces cyclophilin B through the activation of transcription factor 6 in gastric adenocarcinoma cells.

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    Jeong, Kwon; Kim, Kiyoon; Kim, Hunsung; Oh, Yoojung; Kim, Seong-Jin; Jo, Yunhee; Choe, Wonchae

    2015-06-01

    Hypoxia is an important form of physiological stress that induces cell death, due to the resulting endoplasmic reticulum (ER) stress, particularly in solid tumors. Although previous studies have indicated that cyclophilin B (CypB) plays a role in ER stress, there is currently no direct information supporting the mechanism of CypB involvement under hypoxic conditions. However, it has previously been demonstrated that ER stress positively regulates the expression of CypB. In the present study, it was demonstrated that CypB is transcriptionally regulated by hypoxia-mediated activation of transcription factor 6 (ATF6), an ER stress transcription factor. Subsequently, the effects of ATF6 on CypB promoter activity were investigated and an ATF6-responsive region in the promoter was identified. Hypoxia and ATF6 expression each increased CypB promoter activity. Collectively, these results demonstrate that ATF6 positively regulates the expression of CypB by binding to an ATF6-responsive region in the promoter, which may play an important role in the attenuation of apoptosis in the adaption to hypoxia. These results suggest that CypB may be a key molecule in the adaptation of cells to hypoxic conditions.

  16. Neuroprotective effects of overexpressed cyclophilin B against Aβ-induced neurotoxicity in PC12 cells.

    Science.gov (United States)

    Oh, Yoojung; Kim, Eun Young; Kim, Yeonghwan; Jin, Jizi; Jin, Byung Kwan; Jahng, Geon-Ho; Jung, Min Hyung; Park, Chan; Kang, Insug; Ha, Joohun; Choe, Wonchae

    2011-08-15

    Accumulated amyloid-β (Aβ) is a well-known cause of neuronal apoptosis in Alzheimer disease and functions in part by generating oxidative stress. Our previous work suggested that cyclophilin B (CypB) protects against endoplasmic reticulum (ER) stress. Therefore, in this study we examined the ability of CypB to protect against Aβ toxicity. CypB is present in the neurons of rat and mouse brains, and treating neural cells with Aβ(25-35) mediates apoptotic cell death. Aβ(25-35)-induced neuronal toxicity was inhibited by the overexpression of CypB as measured by cell viability, apoptotic morphology, sub-G1 cell population, intracellular reactive oxygen species accumulation, activated caspase-3, PARP cleavage, Bcl-2 proteins, mitogen-activated protein kinase (MAPK) activation, and phosphoinositide 3-kinase (PI-3-K) activation. CypB/R95A PPIase mutants did not reduce Aβ(25-35) toxicity. We showed that Aβ(25-35)-induced apoptosis is more severe in a CypB knockdown model, confirming that CypB protects against Aβ(25-35)-induced toxicity. Consequently, these findings suggest that CypB may protect against Aβ toxicity by its antioxidant properties, by regulating MAPK and PI-3-K signaling, and through the ER stress pathway. Copyright © 2011 Elsevier Inc. All rights reserved.

  17. Cyclophilin B binding to platelets supports calcium-dependent adhesion to collagen.

    Science.gov (United States)

    Allain, F; Durieux, S; Denys, A; Carpentier, M; Spik, G

    1999-08-01

    We have recently reported that cyclophilin B (CyPB), a secreted cyclosporine-binding protein, could bind to T lymphocytes through interactions with two types of binding sites. The first ones, referred to as type I, involve interactions with the conserved domain of CyPB and promote the endocytosis of surface-bound ligand, while the second type of binding sites, termed type II, are represented by glycosaminoglycans (GAG). Here, we further investigated the interactions of CyPB with blood cell populations. In addition to lymphocytes, CyPB was found to interact mainly with platelets. The binding is specific, with a dissociation constant (kd) of 9 +/- 3 nmol/L and the number of sites estimated at 960 +/- 60 per cell. Platelet glycosaminoglycans are not required for the interactions, but the binding is dramatically reduced by active cyclosporine derivatives. We then analyzed the biologic effects of CyPB and found a significant increase in platelet adhesion to collagen. Concurrently, CyPB initiates a transmembranous influx of Ca(2+) and induces the phosphorylation of the P-20 light chains of myosin. Taken together, the present results demonstrate for the first time that extracellular CyPB specifically interacts with platelets through a functional receptor related to the lymphocyte type I binding sites and might act by regulating the activity of a receptor-operated membrane Ca(2+) channel.

  18. Effect of cyclophilin A on gene expression in human pancreatic cancer cells.

    Science.gov (United States)

    Li, Min; Wang, Hao; Li, Fei; Fisher, William E; Chen, Changyi; Yao, Qizhi

    2005-11-01

    We previously found that cyclophilin A (CypA) is overexpressed in human pancreatic cancer cells and stimulates cell proliferation through CD147. In this study, we further investigated the effect of CypA on gene expression of several key molecules that are involved in pancreatic cancer cell proliferation. Human pancreatic cancer cell lines (Panc-1, MIA PaCa-2, and BxPC-3) and human pancreatic ductal epithelial (HPDE) cells were used. The messenger RNA (mRNA) levels of CypA, CypB, CD147, neuropilins (NRPs), vascular endothelial growth factor (VEGF), and VEGF receptors upon the treatment of exogenous recombinant human CypA were determined by real-time reverse-transcription polymerase chain reaction. Exogenous human recombinant CypA reduced the mRNA levels of NRP-1 and VEGF, but not endogenous CypA, CypB, and CD147, in Panc-1, MIA PaCa-2, and BxPC-3 cells. In contrast, HPDE cells showed a decrease of endogenous CypA and CD147 mRNA, but not detectable changes of CypB, NRPs, and VEGF mRNA levels upon exogenous CypA treatment. These data show that exogenous CypA downregulates NRP-1 and VEGF expression in pancreatic cancer cells. This effect is different in normal HPDE cells. Thus, soluble CypA may affect cell growth of pancreatic cancer.

  19. Role of cyclophilin B in tumorigenesis and cisplatin resistance in hepatocellular carcinoma in humans.

    Science.gov (United States)

    Kim, Yeonghwan; Jang, Miran; Lim, Sangbin; Won, Hyeran; Yoon, Kyung-Sik; Park, Jae-Hoon; Kim, Hyo Jong; Kim, Byung-Ho; Park, Won-Sang; Ha, Joohun; Kim, Sung-Soo

    2011-11-01

    Cyclophilin B (CypB) performs diverse roles in living cells, but its role in hepatocellular carcinoma (HCC) is largely unclear. To reveal its role in HCC, we investigated the induction of CypB under hypoxia and its functions in tumor cells in vitro and in vivo. Here, we demonstrated that hypoxia-inducible factor 1α (HIF-1α) induces CypB under hypoxia. Interestingly, CypB protected tumor cells, even p53-defective HCC cells, against hypoxia- and cisplatin-induced apoptosis. Furthermore, it regulated the effects of HIF-1α, including those in angiogenesis and glucose metabolism, via a positive feedback loop with HIF-1α. The tumorigenic and chemoresistant effects of CypB were confirmed in vivo using a xenograft model. Finally, we showed that CypB is overexpressed in 78% and 91% of the human HCC and colon cancer tissues, respectively, and its overexpression in these cancers reduced patient survival. These results indicate that CypB induced by hypoxia stimulates the survival of HCC via a positive feedback loop with HIF-1α, indicating that CypB is a novel candidate target for developing chemotherapeutic agents against HCC and colon cancer. Copyright © 2011 American Association for the Study of Liver Diseases.

  20. Spatiotemporal behavior of nuclear cyclophilin B indicates a role in RNA transcription.

    Science.gov (United States)

    Dieriks, Birger; Van Oostveldt, Patrick

    2012-06-01

    Cyclophilin B (CypB) is an ubiquitously expressed protein, which performs several intra- and extracellular functions. Despite its abundant use as a household protein, little is known about its exact cellular localization and dynamics. In the present study we show that endogenous CypB localizes in one of two distinct compartments, either within the endoplasmic reticulum (ER) or inside the nucleus, accumulating in the fibrillar centers of the nucleoli. By means of a genetic deletion screen, we identified a minimal nucleolar localization signal for efficient relocation to the nucleoli. Within the fibrillar centers, CypB colocalized with RNA polymerase, upstream binding factor-1 (UBF), fibrillarin and dyskerin (DCK1). Even after chemical disruption of the nucleoli, a strong interaction with these proteins remained. Using live cell imaging, we showed a persistent colocalization of CypB with proteins involved in the ribosome biogenesis during the transcriptionally more active phases of the cell cycle. Supported by in silico data, our observations suggest that CypB interacts with these proteins and is involved in ribosome biogenesis and RNA transcription.

  1. A BAX/BAK and cyclophilin D-independent intrinsic apoptosis pathway.

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    Sebastián Zamorano

    Full Text Available Most intrinsic death signals converge into the activation of pro-apoptotic BCL-2 family members BAX and BAK at the mitochondria, resulting in the release of cytochrome c and apoptosome activation. Chronic endoplasmic reticulum (ER stress leads to apoptosis through the upregulation of a subset of pro-apoptotic BH3-only proteins, activating BAX and BAK at the mitochondria. Here we provide evidence indicating that the full resistance of BAX and BAK double deficient (DKO cells to ER stress is reverted by stimulation in combination with mild serum withdrawal. Cell death under these conditions was characterized by the appearance of classical apoptosis markers, caspase-9 activation, release of cytochrome c, and was inhibited by knocking down caspase-9, but insensitive to BCL-X(L overexpression. Similarly, the resistance of BIM and PUMA double deficient cells to ER stress was reverted by mild serum withdrawal. Surprisingly, BAX/BAK-independent cell death did not require Cyclophilin D (CypD expression, an important regulator of the mitochondrial permeability transition pore. Our results suggest the existence of an alternative intrinsic apoptosis pathway emerging from a cross talk between the ER and the mitochondria.

  2. Pharmacological Cyclophilin Inhibitors Prevent Intoxication of Mammalian Cells with Bordetella pertussis Toxin.

    Science.gov (United States)

    Ernst, Katharina; Eberhardt, Nina; Mittler, Ann-Katrin; Sonnabend, Michael; Anastasia, Anna; Freisinger, Simon; Schiene-Fischer, Cordelia; Malešević, Miroslav; Barth, Holger

    2018-05-01

    The Bordetella pertussis toxin (PT) is one important virulence factor causing the severe childhood disease whooping cough which still accounted for approximately 63,000 deaths worldwide in children in 2013. PT consists of PTS1, the enzymatically active (A) subunit and a non-covalently linked pentameric binding/transport (B) subunit. After endocytosis, PT takes a retrograde route to the endoplasmic reticulum (ER), where PTS1 is released into the cytosol. In the cytosol, PTS1 ADP-ribosylates inhibitory alpha subunits of trimeric GTP-binding proteins (Giα) leading to increased cAMP levels and disturbed signalling. Here, we show that the cyclophilin (Cyp) isoforms CypA and Cyp40 directly interact with PTS1 in vitro and that Cyp inhibitors cyclosporine A (CsA) and its tailored non-immunosuppressive derivative VK112 both inhibit intoxication of CHO-K1 cells with PT, as analysed in a morphology-based assay. Moreover, in cells treated with PT in the presence of CsA, the amount of ADP-ribosylated Giα was significantly reduced and less PTS1 was detected in the cytosol compared to cells treated with PT only. The results suggest that the uptake of PTS1 into the cytosol requires Cyps. Therefore, CsA/VK112 represent promising candidates for novel therapeutic strategies acting on the toxin level to prevent the severe, life-threatening symptoms caused by PT.

  3. Interaction with glycosaminoglycans is required for cyclophilin B to trigger integrin-mediated adhesion of peripheral blood T lymphocytes to extracellular matrix

    OpenAIRE

    Allain, Fabrice; Vanpouille, Christophe; Carpentier, Mathieu; Slomianny, Marie-Christine; Durieux, Sandrine; Spik, Geneviève

    2002-01-01

    Cyclophilins A and B (CyPA and CyPB) are cyclosporin A-binding proteins that are involved in inflammatory events. We have reported that CyPB interacts with two types of cell-surface-binding sites. The first site corresponds to a functional receptor and requires interaction with the central core of CyPB. This region is highly conserved in cyclophilins, suggesting that CyPA and CyPB might share biological activities mediated by interaction with this receptor. The second site is identified with ...

  4. Seroprevalence of Toxoplasma gondii infection in goats from the south-west region of Poland and the detection of T. gondii DNA in goat milk.

    Science.gov (United States)

    Sroka, Jacek; Kusyk, Pawel; Bilska-Zajac, Ewa; Karamon, Jacek; Dutkiewicz, Jacek; Wojcik-Fatla, Angelina; Zajac, Violetta; Stojecki, Krzysztof; Rozycki, Miroslaw; Cencek, Tomasz

    2017-07-11

    Toxoplasma gondii (Nicolle et Manceaux, 1908) is an obligatory intracellular protozoan parasite prevalent in animals and humans worldwide having medical and veterinary importance on account of causing abortion or congenital disease in intermediate hosts, including man. Since T. gondii has already been identified in the milk of goats, Capra aegagrus hircus (Linnaeus), the possibility of acquiring infection by ingesting unpasteurised goat milk should be taken into consideration. Thus, the aim of the present study was to determine the presence of T. gondii DNA in goat milk. First, 73 goats (females) from 36 farms located in Poland were examined serologically by direct agglutination test (DAT) to estimate the T. gondii serological status. Milk samples from 60 selected lactating females were examined for the presence of T. gondii DNA by Real time PCR and nested PCR (B1 gene). To estimate the clonal type of detected T. gondii, multiplex PCR was performed using 6 markers. In DAT, positive results were found in 70% of 73 goats. Among examined 60 milk samples, 65% were positive in Real time PCR and 43% in nested PCR. It is noteworthy that 11 samples positive in PCR were collected from seronegative goats. The multilocus PCR analysis mostly revealed the occurrence of genotype III, which is relatively rare in Europe. The recorded high prevalence of anti-Toxoplasma antibodies in tested goats (70%), associated with a high prevalence of T. gondii DNA in goat milk samples (65%), indicates a potential risk of the parasite transmission through goat milk ingestion.

  5. First report of Toxoplasma gondii seroprevalence in peafowls in Yunnan Province, Southwestern China

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    Tian Yi-Ming

    2012-09-01

    Full Text Available Abstract Background Toxoplasma gondii is an intracellular protozoan parasite infecting almost all warm-blooded animals, including birds, with a worldwide distribution. Surveys of T. gondii infection in wild birds have been reported extensively in the world, but little is known of T. gondii infection in peafowls worldwide. This study was performed to determine the seroprevalence of T. gondii infection in peafowls in Yunnan Province, southwestern China. Methods Sera from 277 peafowls, including 272 blue peafowls (Pavo cristatus and 5 green peafowls (Pavo muticus originated from two geographic areas in Yunnan Province were assayed for T. gondii antibodies using the modified agglutination test (MAT. Results Specific T. gondii antibodies were detected in 35 of 277 (12.64% peafowls (MAT titer ≥ 1:5. Seropositive birds were found in both species, 33 in 272 blue peafowls and 2 in 5 green peafowls. There was no significant difference in T. gondii seroprevalence between the adolescent birds (6.74% and the adult birds (6.67% (P > 0.05. The geographical origins of peafowls was found to be highly associated with T. gondii infection in the present study, a statistically significant difference in T. gondii seropositivity was observed between peafowls from Kunming (31.08% and those from Xishuangbanna Dai Autonomous Prefecture (5.91% (OR = 10.956, 95% CI = 1.632-73.545, P = 0.014. Statistical analyses showed that there were no significant interactions between ages and geographical origins of peafowls (P > 0.05. Conclusions The results of the present survey indicated that infection of peafowls with T. gondii is widespread in Yunnan Province, which has significant public health concerns and implications for prevention and control of toxoplamosis in this province. To our knowledge, this is the first seroprevalence report of T. gondii infection in China’s southwestern Yunnan Province.

  6. First report of Toxoplasma gondii seroprevalence in peafowls in Yunnan Province, Southwestern China.

    Science.gov (United States)

    Tian, Yi-Ming; Dai, Fei-Yan; Huang, Si-Yang; Deng, Zu-Hong; Duan, Gang; Zhou, Dong-Hui; Yang, Jian-Fa; Weng, Ya-Biao; Zhu, Xing-Quan; Zou, Feng-Cai

    2012-09-19

    Toxoplasma gondii is an intracellular protozoan parasite infecting almost all warm-blooded animals, including birds, with a worldwide distribution. Surveys of T. gondii infection in wild birds have been reported extensively in the world, but little is known of T. gondii infection in peafowls worldwide. This study was performed to determine the seroprevalence of T. gondii infection in peafowls in Yunnan Province, southwestern China. Sera from 277 peafowls, including 272 blue peafowls (Pavo cristatus) and 5 green peafowls (Pavo muticus) originated from two geographic areas in Yunnan Province were assayed for T. gondii antibodies using the modified agglutination test (MAT). Specific T. gondii antibodies were detected in 35 of 277 (12.64%) peafowls (MAT titer ≥ 1:5). Seropositive birds were found in both species, 33 in 272 blue peafowls and 2 in 5 green peafowls. There was no significant difference in T. gondii seroprevalence between the adolescent birds (6.74%) and the adult birds (6.67%) (P > 0.05). The geographical origins of peafowls was found to be highly associated with T. gondii infection in the present study, a statistically significant difference in T. gondii seropositivity was observed between peafowls from Kunming (31.08%) and those from Xishuangbanna Dai Autonomous Prefecture (5.91%) (OR = 10.956, 95% CI = 1.632-73.545, P = 0.014). Statistical analyses showed that there were no significant interactions between ages and geographical origins of peafowls (P > 0.05). The results of the present survey indicated that infection of peafowls with T. gondii is widespread in Yunnan Province, which has significant public health concerns and implications for prevention and control of toxoplamosis in this province. To our knowledge, this is the first seroprevalence report of T. gondii infection in China's southwestern Yunnan Province.

  7. Isolation and characterization of new genetic types of toxoplasma gondii and prevalence of trichinella murrelli from black bear (Ursus americanus)

    Science.gov (United States)

    Black bears (Ursus americanus) are hosts for two important zoonotic parasites, Toxoplasma gondii and Trichinella spp. and bears are hunted for human consumption in the USA. Little is known of the genetic diversity of T. gondii circulating in wildlife. In the present study, antibodies to T. gondii we...

  8. Toxoplasma gondii peptide ligands open the gate of the HLA class I binding groove

    DEFF Research Database (Denmark)

    McMurtrey, Curtis; Trolle, Thomas; Sansom, Tiffany

    2016-01-01

    HLA class I presentation of pathogen-derived peptide ligands is essential for CD8+ T cell recognition of Toxoplasma gondii infected cells. Currently, little data exist pertaining to peptides that are presented after T. gondii infection. Herein we purify HLA-A*02:01 complexes from T. gondii infect...

  9. High prevalence of toxoplasmosis in cats from Egypt: isolation of viable Toxoplasma Gondii, tissue distribution, and isolate designation

    Science.gov (United States)

    Cats are important in the epidemiology of Toxoplasma gondii because they are the only hosts that excrete environmentally resistant oocysts in feces. In the present study, 158 feral cats from Giza, Egypt were examined for T. gondii infection. Antibodies to T. gondii were found in 97.4% with modified ...

  10. Long-Term Protective Immune Response Elicited by Vaccination with an Expression Genomic Library of Toxoplasma gondii

    OpenAIRE

    Fachado, Alberto; Rodriguez, Alexandro; Molina, Judith; Silvério, Jaline C.; Marino, Ana P. M. P.; Pinto, Luzia M. O.; Angel, Sergio O.; Infante, Juan F.; Traub-Cseko, Yara; Amendoeira, Regina R.; Lannes-Vieira, Joseli

    2003-01-01

    Immunization of BALB/c mice with an expression genomic library of Toxoplasma gondii induces a Th1-type immune response, with recognition of several T. gondii proteins (21 to 117 kDa) and long-term protective immunity against a lethal challenge. These results support further investigations to achieve a multicomponent anti-T. gondii DNA vaccine.

  11. Prevalence of Toxoplasma gondii in native donkeys in Mosul

    Directory of Open Access Journals (Sweden)

    Kh. J. Hussain

    2011-01-01

    Full Text Available The aim of the study was to determine the prevalence of Toxoplasma gondii in native donkeys in Mousl, Iraq. Fifty two sera (9 males and 43 females were examined by Latex agglutination test, Modified latex agglutination test with 2- mercaptoethanol test and Indirect ELISA test (Indirect IgG ELISA. The prevalence of Toxoplasma gondii in native donkeys was 46.15 %. Acute cases 8.33% and chronic cases 91.67 % when differentiated by Modified latex agglutination test with 2- mercaptoethanol test. The percentages of female and male infections were 51.16% (22/43 and 22.22% (2/9, respectively by using latex agglutination test, and the titeration of antibodies ranged between 1:20 - 1:1280 and for Indirect IgG ELISA it was 22.72% positive cases.

  12. Implication of the Whitefly Bemisia tabaci Cyclophilin B Protein in the Transmission of Tomato yellow leaf curl virus.

    Science.gov (United States)

    Kanakala, Surapathrudu; Ghanim, Murad

    2016-01-01

    Tomato yellow leaf curl virus (TYLCV) is a single-stranded (ssDNA) begomoviruses that causes severe damage to tomato and several other crops worldwide. TYLCV is exclusively transmitted by the sweetpotato whitefly, Bemisia tabaci in a persistent circulative and propagative manner. Previous studies have shown that the transmission, retention, and circulation of TYLCV in its vector involves interaction with insect and endosymbiont proteins, which aid in the transmission of the virus, or have a protective role in response to the presence of the virus in the insect body. However, only a low number of such proteins have been identified. Here, the role of B. tabaci Cyclophilin B (CypB) in the transmission of TYLCV protein was investigated. Cyclophilins are a large family of cellular prolyl isomerases that have many molecular roles including facilitating protein-protein interactions in the cell. One cyclophilin protein has been implicated in aphid-luteovirus interactions. We demonstrate that the expression of CypB from B. tabaci is altered upon TYLCV acquisition and retention. Further experiments used immunocapture-PCR and co-immunolocalization and demonstrated a specific interaction and colocalization between CypB and TYLCV in the the midgut, eggs, and salivary glands. Membrane feeding of anti-CypB antibodies and TYLCV-infected plants showed a decrease in TYLCV transmission, suggesting a critical role that CypB plays in TYLCV transmission. Further experiments, which used membrane feeding with the CypB inhibitor Cyclosporin A showed decrease in CypB-TYLCV colocalization in the midgut and virus transmission. Altogether, our results indicate that CypB plays an important role in TYLCV transmission by B. tabaci .

  13. High level expression and characterization of the cyclophilin B gene from the anaerobic fungus Orpinomyces sp. strain PC-2.

    Science.gov (United States)

    Chen, Huizhong; Li, Xin-Liang; Xu, Haiyan; Ljungdahl, Lars G; Cerniglia, Carl E

    2006-01-01

    Cyclophilins are an evolutionarily conserved family of peptidyl-prolyl cis-trans isomerases (PPIases). A cyclophilin B (cypB) gene from the anaerobic fungus Orpinomyces sp. strain PC-2 was cloned and overexpressed in Escherichia coli. It was expressed as an amino-terminal 6 x His-tagged recombinant protein to facilitate purification. Highly purified protein (26.5 kDa) was isolated by two chromatographic steps involving affinity and gel filtration for biochemical studies of the enzyme. The recombinant CypB displayed PPIase activity with a k(cat)/K(m) of 8.9 x 10(6) M(-1) s(-1) at 10 degrees C and pH 7.8. It was inhibited by cyclosporin A (CsA) with an IC(50) of 23.5 nM, similar to those of the native protein and other cyclophilin B enzymes from animals. Genomic DNA analysis of cypB revealed that it was present as a single copy in Orpinomyces PC-2 and contained two introns, indicating it has a eukaryotic origin. It is one of the most heavily interrupted genes with intron sequences found in anaerobic fungi. The three-dimensional model of Orpinomyces PC-2 CypB was predicted with a homology modeling approach using the Swiss-Model Protein Modeling Server and three dimensional structure of human CypB as a template. The overall architecture of the CypB molecule is very similar to that of human CypB.

  14. Implication of the whitefly Bemisia tabaci Cyclophilin B protein in the transmission of Tomato yellow leaf curl virus

    Directory of Open Access Journals (Sweden)

    Surapathrudu Kanakala

    2016-11-01

    Full Text Available Tomato yellow leaf curl virus (TYLCV is a single-stranded (ssDNA begomoviruses that causes severe damage to tomato and several other crops worldwide. TYLCV is exclusively transmitted by the sweetpotato whitefly, Bemisia tabaci in a persistent circulative and propagative manner. Previous studies have shown that the transmission, retention and circulation of TYLCV in its vector involves interaction with insect and endosymbiont proteins, which aid in the transmission of the virus, or have a protective role in response to the presence of the virus in the insect body. However, only a low number of such proteins have been identified. Here, the role of B. tabaci Cyclophilin B (CypB in the transmission of TYLCV protein was investigated. Cyclophilins (Cyps are a large family of cellular prolyl isomerases that have many molecular roles including facilitating protein-protein interactions in the cell. One cyclophilin protein has been implicated in aphid-luteovirus interactions. We demonstrate that the expression of CypB from B. tabaci is altered upon TYLCV acquisition and retention. Further experiments used immunocapture-PCR and co-immunolocalization and demonstrated a specific interaction and colocalization between CypB and TYLCV in the the midgut, eggs and salivary glands. Membrane feeding of anti-CypB antibodies and TYLCV infected plants showed a decrease in TYLCV transmission, suggesting a critical role that CypB plays in TYLCV transmission. Further experiments, which used membrane feeding with the CypB inhibitor Cyclosporin A (CsA showed decrease in CypB-TYLCV colocalization in the midgut and virus transmission. Altogether, our results indicate that CypB plays an important role in TYLCV transmission by B. tabaci.

  15. Risk factors for Toxoplasma gondii infection in Kohat District, Pakistan

    OpenAIRE

    Gul Naila; Zareen Shehzad; Ur Rehman Faiz; Ur Rehman Hameed; Qayyum Sumbel; Khan Sumiya; Khan Feroz; Ali Khan Munsif; Saeed Kausar; Ayub Abid; Hayat Azam; Ateeq Muhammad; Ahmad Waqar

    2017-01-01

    Toxoplasma gondii is a widespread zoonotic parasite that is the causative agent for toxoplasmosis in human and completes its life cycle in separate hosts. Considering the significance of the infection, the current study was designed to asses to various risk factors for the parasite transmission to human in Kohat District, Pakistan. A total of 122 suspected individuals were asked to fill pre-designed questionnaire. A total of 44 (36.07%) individuals were found to be infected wit...

  16. Toxoplasma gondii actively inhibits neuronal function in chronically infected mice.

    Directory of Open Access Journals (Sweden)

    Fahad Haroon

    Full Text Available Upon infection with the obligate intracellular parasite Toxoplasma gondii, fast replicating tachyzoites infect a broad spectrum of host cells including neurons. Under the pressure of the immune response, tachyzoites convert into slow-replicating bradyzoites, which persist as cysts in neurons. Currently, it is unclear whether T. gondii alters the functional activity of neurons, which may contribute to altered behaviour of T. gondii-infected mice and men. In the present study we demonstrate that upon oral infection with T. gondii cysts, chronically infected BALB/c mice lost over time their natural fear against cat urine which was paralleled by the persistence of the parasite in brain regions affecting behaviour and odor perception. Detailed immunohistochemistry showed that in infected neurons not only parasitic cysts but also the host cell cytoplasm and some axons stained positive for Toxoplasma antigen suggesting that parasitic proteins might directly interfere with neuronal function. In fact, in vitro live cell calcium (Ca(2+ imaging studies revealed that tachyzoites actively manipulated Ca(2+ signalling upon glutamate stimulation leading either to hyper- or hypo-responsive neurons. Experiments with the endoplasmatic reticulum Ca(2+ uptake inhibitor thapsigargin indicate that tachyzoites deplete Ca(2+ stores in the endoplasmatic reticulum. Furthermore in vivo studies revealed that the activity-dependent uptake of the potassium analogue thallium was reduced in cyst harbouring neurons indicating their functional impairment. The percentage of non-functional neurons increased over time In conclusion, both bradyzoites and tachyzoites functionally silence infected neurons, which may significantly contribute to the altered behaviour of the host.

  17. Genotyping of polymorphic effectors of Toxoplasma gondii isolates from China

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    Weisheng Cheng

    2017-11-01

    Full Text Available Abstract Background Toxoplasma gondii is an opportunistic protozoan apicomplexan and obligate intracellular parasite that infects a wide range of animals and humans. Rhoptry proteins 5 (ROP5, ROP16, ROP18 and dense granules 15 (GRA15 are the important effectors secreted by T. gondii which link to the strain virulence for mice and modulate the host’s response to the parasite. Little has been known about these molecules as well as GRA3 in type Chinese 1 strains that show polymorphism among strains of archetypical genotypes. This study examined the genetic diversity of these effectors and its correlated virulence in mice among T. gondii isolates from China. Results Twenty-one isolates from stray cats were detected, of which 15 belong to Chinese 1, and 6 to ToxoDB #205. Wh6 isolate, a Chinese 1 strain, has an avirulent phenotype. PCR-RFLP results of ROP5 and ROP18 presented few variations among the strains. Genotyping of GRA15 and ROP16 revealed that all the strains belong to type II allele except Xz7 which carries type I allele. ROP16 amino acid alignment at 503 locus demonstrated that 17 isolates are featured as type I or type III (ROP16I/III, and the other 4 as type II (ROP16II. The strains investigated may be divided into four groups based on GRA3 amino acid alignment, and all isolates of type Chinese 1 belong to the μ-1 allele except Wh6 which is identical to type II strain. Conclusions PCR-RFLP and sequence alignment analyses of ROP5, ROP16, ROP18, GRA3, and GRA15 in T. gondii revealed that strains with the same genotype may have variations in some of their key genes. GRA3 variation exhibited by Wh6 strain may be associated with the difference in phenotype and pathogenesis.

  18. Global Analysis of Palmitoylated Proteins in Toxoplasma gondii.

    Science.gov (United States)

    Foe, Ian T; Child, Matthew A; Majmudar, Jaimeen D; Krishnamurthy, Shruthi; van der Linden, Wouter A; Ward, Gary E; Martin, Brent R; Bogyo, Matthew

    2015-10-14

    Post-translational modifications (PTMs) such as palmitoylation are critical for the lytic cycle of the protozoan parasite Toxoplasma gondii. While palmitoylation is involved in invasion, motility, and cell morphology, the proteins that utilize this PTM remain largely unknown. Using a chemical proteomic approach, we report a comprehensive analysis of palmitoylated proteins in T. gondii, identifying a total of 282 proteins, including cytosolic, membrane-associated, and transmembrane proteins. From this large set of palmitoylated targets, we validate palmitoylation of proteins involved in motility (myosin light chain 1, myosin A), cell morphology (PhIL1), and host cell invasion (apical membrane antigen 1, AMA1). Further studies reveal that blocking AMA1 palmitoylation enhances the release of AMA1 and other invasion-related proteins from apical secretory organelles, suggesting a previously unrecognized role for AMA1. These findings suggest that palmitoylation is ubiquitous throughout the T. gondii proteome and reveal insights into the biology of this important human pathogen. Copyright © 2015 Elsevier Inc. All rights reserved.

  19. The neurotropic parasite Toxoplasma gondii increases dopamine metabolism.

    Directory of Open Access Journals (Sweden)

    Emese Prandovszky

    Full Text Available The highly prevalent parasite Toxoplasma gondii manipulates its host's behavior. In infected rodents, the behavioral changes increase the likelihood that the parasite will be transmitted back to its definitive cat host, an essential step in completion of the parasite's life cycle. The mechanism(s responsible for behavioral changes in the host is unknown but two lines of published evidence suggest that the parasite alters neurotransmitter signal transduction: the disruption of the parasite-induced behavioral changes with medications used to treat psychiatric disease (specifically dopamine antagonists and identification of a tyrosine hydroxylase encoded in the parasite genome. In this study, infection of mammalian dopaminergic cells with T. gondii enhanced the levels of K+-induced release of dopamine several-fold, with a direct correlation between the number of infected cells and the quantity of dopamine released. Immunostaining brain sections of infected mice with dopamine antibody showed intense staining of encysted parasites. Based on these analyses, T. gondii orchestrates a significant increase in dopamine metabolism in neural cells. Tyrosine hydroxylase, the rate-limiting enzyme for dopamine synthesis, was also found in intracellular tissue cysts in brain tissue with antibodies specific for the parasite-encoded tyrosine hydroxylase. These observations provide a mechanism for parasite-induced behavioral changes. The observed effects on dopamine metabolism could also be relevant in interpreting reports of psychobehavioral changes in toxoplasmosis-infected humans.

  20. Latent Toxoplasma gondii infection leads to improved action control.

    Science.gov (United States)

    Stock, Ann-Kathrin; Heintschel von Heinegg, Evelyn; Köhling, Hedda-Luise; Beste, Christian

    2014-03-01

    The parasite Toxoplasma gondii has been found to manipulate the behavior of its secondary hosts to increase its own dissemination which is commonly believed to be to the detriment of the host (manipulation hypothesis). The manipulation correlates with an up-regulation of dopaminergic neurotransmission. In humans, different pathologies have been associated with T. gondii infections but most latently infected humans do not seem to display overt impairments. Since a dopamine plus does not necessarily bear exclusively negative consequences in humans, we investigated potential positive consequences of latent toxoplasmosis (and the presumed boosting of dopaminergic neurotransmission) on human cognition and behavior. For this purpose, we focused on action cascading which has been shown to be modulated by dopamine. Based on behavioral and neurophysiological (EEG) data obtained by means of a stop-change paradigm, we were able to demonstrate that healthy young humans can actually benefit from latent T. gondii infection as regards their performance in this task (as indicated by faster response times and a smaller P3 component). The data shows that a latent infection which is assumed to affect the dopaminergic system can lead to paradoxical improvements of cognitive control processes in humans. Copyright © 2013 Elsevier Inc. All rights reserved.

  1. The Ultrastructural Effects of Sulfachloropyrazine on Toxoplasma Gondii Tachyzoites

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    YB Zeng

    2013-03-01

    Full Text Available Background: Toxoplasmosis is one of the most common parasitic infections of humans and other mammals. This study was aimed to understand the mechanism of action of veterinary medicine-sulfachlo­ropyrazine (SPZ, 99.97% against Toxoplasma gondii.Methods: T. gondii tachyzoites were soaked in PBS (as a control or SPZ (250 mg/mL for 2 h at 37 °C. After being processed, any ultrastructural changes of the tachyzoites that had occurred were observed by Scanning Electron Microscopy (SEM and Transmission Electron Microscopy (TEM.Results: The tachyzoites from control groups with a uniform size had a smooth surface and intact cell or nuclear membranes. In addition, an oval-shaped nucleus, conoids and micronemes were also observed. By contrast, many parasites from the SPZ-treated groups were detrimentally affected by the treatment. Some appeared to be of the vacuolization in their cytoplasm, with the substantial reduc­tion in the number of dense granules and the blur of some organelles.Conclusion: The morphology and ultrastructure of tachyzoites can be affected significantly by SPZ, which might kill the parasite by inhibiting its energy metabolism, inducing apoptosis and damaging its structure. The study provides an experimental basis for further study on the mechanism of SPZ against T. gondii.

  2. Seroprevalence of Toxoplasma gondii infection among pregnant women in Cameroon

    Directory of Open Access Journals (Sweden)

    Anna L. Njunda

    2011-09-01

    Full Text Available Toxoplasmosis is caused by an intracellular protozoan, Toxoplasma gondii, which has a wide geographical distribution. The congenital form results in a gestational form that can present a temporary parasiteamia that will infect the fetus. For this reason early diagnosis in pregnancy is highly desirable, allowing prompt intervention in cases of infection. The aim of this study was to determine the seroprevalence of Toxoplasma gondii antibodies among pregnant women attending the Douala General Hospital. The study was carried out between March and July 2009, whereby 110 pregnant women were tested for IgG and IgM antibodies and information about eating habits and hygienic conditions was collected using a questionnaire. These women’s ages ranged from 20-44 years old with an average of 29.9 years; the overall IgG and IgM seroprevalence was 70% and 2.73 % respectively. Seroprevalence was significantly high amongst women who ate raw vegetables (76.39%, P<0.05 and there was a significant trend towards a higher seroprevalence in women who did not have a good source of water (75.58%, P<0.05. This research showed that consumption raw vegetables and poor quality drinking water are two risk factors associated with Toxoplasma gondii infection amongst pregnant women attending the Douala General Hospital in Cameroon.

  3. Genomic and proteomic analysis of Schizaphis graminum reveals cyclophilin proteins are involved in the transmission of cereal yellow dwarf virus.

    Directory of Open Access Journals (Sweden)

    Cecilia Tamborindeguy

    Full Text Available Yellow dwarf viruses cause the most economically important virus diseases of cereal crops worldwide and are transmitted by aphid vectors. The identification of aphid genes and proteins mediating virus transmission is critical to develop agriculturally sustainable virus management practices and to understand viral strategies for circulative movement in all insect vectors. Two cyclophilin B proteins, S28 and S29, were identified previously in populations of Schizaphisgraminum that differed in their ability to transmit the RPV strain of Cereal yellow dwarf virus (CYDV-RPV. The presence of S29 was correlated with F2 genotypes that were efficient virus transmitters. The present study revealed the two proteins were isoforms, and a single amino acid change distinguished S28 and S29. The distribution of the two alleles was determined in 12 F2 genotypes segregating for CYDV-RPV transmission capacity and in 11 genetically independent, field-collected S. graminum biotypes. Transmission efficiency for CYDV-RPV was determined in all genotypes and biotypes. The S29 isoform was present in all genotypes or biotypes that efficiently transmit CYDV-RPV and more specifically in genotypes that efficiently transport virus across the hindgut. We confirmed a direct interaction between CYDV-RPV and both S28 and S29 using purified virus and bacterially expressed, his-tagged S28 and S29 proteins. Importantly, S29 failed to interact with a closely related virus that is transported across the aphid midgut. We tested for in vivo interactions using an aphid-virus co-immunoprecipitation strategy coupled with a bottom-up LC-MS/MS analysis using a Q Exactive mass spectrometer. This analysis enabled us to identify a third cyclophilin protein, cyclophilin A, interacting directly or in complex with purified CYDV-RPV. Taken together, these data provide evidence that both cyclophilin A and B interact with CYDV-RPV, and these interactions may be important but not sufficient to mediate

  4. Cyclophilin B as a co-regulator of prolactin-induced gene expression and function in breast cancer cells

    OpenAIRE

    Fang, Feng; Zheng, Jiamao; Galbaugh, Traci L; Fiorillo, Alyson A; Hjort, Elizabeth E; Zeng, Xianke; Clevenger, Charles V

    2010-01-01

    The effects of prolactin (PRL) during the pathogenesis of breast cancer are mediated in part though Stat5 activity enhanced by its interaction with its transcriptional inducer, the prolyl isomerase cyclophilin B (CypB). We have demonstrated that knockdown of CypB decreases cell growth, proliferation, and migration, and CypB expression is associated with malignant progression of breast cancer. In this study, we examined the effect of CypB knockdown on PRL signaling in breast cancer cells. CypB...

  5. Expression of Cyclophilin B is Associated with Malignant Progression and Regulation of Genes Implicated in the Pathogenesis of Breast Cancer

    OpenAIRE

    Fang, Feng; Flegler, Ayanna J.; Du, Pan; Lin, Simon; Clevenger, Charles V.

    2009-01-01

    Cyclophilin B (CypB) is a 21-kDa protein with peptidyl-prolyl cis-trans isomerase activity that functions as a transcriptional inducer for Stat5 and as a ligand for CD147. To better understand the global function of CypB in breast cancer, T47D cells with a small interfering RNA-mediated knockdown of CypB were generated. Subsequent expression profiling analysis showed that 663 transcripts were regulated by CypB knockdown, and that many of these gene products contributed to cell proliferation, ...

  6. Involvement of two classes of binding sites in the interactions of cyclophilin B with peripheral blood T-lymphocytes.

    OpenAIRE

    Denys, A; Allain, F; Carpentier, M; Spik, G

    1998-01-01

    Cyclophilin B (CyPB) is a cyclosporin A (CsA)-binding protein, mainly associated with the secretory pathway, and is released in biological fluids. We recently reported that CyPB specifically binds to T-lymphocytes and promotes enhanced incorporation of CsA. The interactions with cellular binding sites involved, at least in part, the specific N-terminal extension of the protein. In this study, we intended to specify further the nature of the CyPB-binding sites on peripheral blood T-lymphocytes...

  7. Sequence variation in TgROP7 gene among Toxoplasma gondii ...

    African Journals Online (AJOL)

    Yomi

    2012-03-27

    Mar 27, 2012 ... Toxoplasma gondii can infect a wide range of hosts including mammals and birds, causing toxoplasmosis which is one of the most common parasitic zoonoses worldwide. The present study examined sequence variation in rhoptry 7 (ROP7) gene among different T. gondii isolates from different hosts and ...

  8. Sporulation and survival of Toxoplasma gondii oocysts in different types of commercial cat litters

    Science.gov (United States)

    Toxoplasma gondii oocysts are environmentally resistant and can survive outdoors for months in the dry and cold climates. In the present study, sporulation and survival of T. gondii oocysts was studied in different types of cat litters commercially available in the US. Oocysts sporulated within 2-...

  9. Role of NETs in the difference in host susceptibility to Toxoplasma gondii between sheep and cattle.

    Science.gov (United States)

    Yildiz, Kader; Gokpinar, Sami; Gazyagci, Aycan Nuriye; Babur, Cahit; Sursal, Neslihan; Azkur, Ahmet Kursat

    2017-07-01

    The main aim of this study was to compare extracellular traps (NETs) formation by polymorphonuclear neutrophils (PMNs) of cattle and sheep when exposed to T. gondii tachyzoites in vitro. The effects of parasite concentrations and different incubation periods on NETs development in cattle and sheep PMNs were studied. The effect of NET structures on host cell invasion by tachyzoites was also studied. This is the first report of NETs development by sheep and cattle PMNs against T. gondii in vitro. T. gondii-induced extracellular DNA production from PMNs was dependent on tachyzoite concentrations and incubation time in both sheep and cattle. Many nuclear and cytoplasmic changes were observed in sheep and cattle PMNs after exposure to T. gondii tachyzoites. The typical appearance of NETs, with MPO, NE and histone (H3) attached to extracellular DNA, was observed. Tachyzoites were entrapped within this structure. Myeloperoxidase (MPO) activity was higher in the cattle PMN-tachyzoite co-cultures than sheep. NETs structures released from sheep PMNs caused mechanical immobilisation of T. gondii tachyzoites, however, NET structures released from cattle PMNs may be lethal to tachyzoites. Bovine MPO may have a lethal effect on T. gondii tachyzoites in vitro during a 3h incubation. Besides other mechanisms that effect on host susceptibility to T. gondii in sheep and cattle, extracellular traps formation as a part of immunological reactions may be play a role in host susceptibility to T. gondii. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Toxoplasmosis in dogs: First report of Toxoplasma gondii infection in any animal species in Angola

    Science.gov (United States)

    Despite the worldwide importance of zoonotic parasite Toxoplasma gondii nothing is known of toxoplasmosis in animals in Angola. The present study aimed at estimating the seroprevalence and also assessing correlates of T. gondii infection in pet dogs from Luanda, Angola. Dogs (n = 103) brought to a v...

  11. A quantitative microbial risk assessment for meatborne Toxoplasma gondii infection in The Netherlands

    NARCIS (Netherlands)

    Opsteegh, M.; Prickaerts, S.; Frankena, K.; Evers, E.G.

    2011-01-01

    Toxoplasma gondii is an important foodborne pathogen, and the cause of a high disease burden due to congenital toxoplasmosis in The Netherlands. The aim of this study was to quantify the relative contribution of sheep, beef and pork products to human T. gondii infections by Quantitative Microbial

  12. Design of a Risk based Control System for Toxoplasma gondii in a pork supply chain

    NARCIS (Netherlands)

    Heres, L.; Swanenburg, M.; Koeijer, de A.A.; Boender, G.J.; Oorburg, D.; Urlings, B.; Wisselink, H.J.

    2015-01-01

    Toxoplasma gondii is recognized as one of the major foodborne pathogens. The European Food Safety Authority (EFSA) concluded that T. gondii is one of the public health hazards in pigs to be covered within a modern meat inspection and advised to include serological testing of pigs and audits of pig

  13. Inactivation of Toxoplasma gondii on blueberries using low dose irradiation without affecting quality

    Science.gov (United States)

    Toxoplasma gondii is a common protozoan parasite, whose environmentally-resistant stage, the oocyst, can contaminate irrigation water and fresh edible produce. Current washing steps in produce processing may not be effective for eliminating T. gondii from at-risk varieties of produce. The objective ...

  14. The first isolation and molecular characterization of Toxoplasma gondii from horses in Serbia.

    NARCIS (Netherlands)

    Klun, Ivana; Uzelac, Aleksandra; Villena, Isabelle; Mercier, Aurélien; Bobić, Branko; Nikolić, Aleksandra; Rajnpreht, Irena; Opsteegh, Marieke; Aubert, Dominique; Blaga, Radu; van der Giessen, Joke; Djurković-Djaković, Olgica

    2017-01-01

    Consumption of undercooked or insufficiently cured meat is a major risk factor for human infection with Toxoplasma gondii. Although horsemeat is typically consumed rare or undercooked, information on the risk of T. gondii from infected horse meat to humans is scarce. Here, we present the results of

  15. "Latent" infection with Toxoplasma gondii: association with trait aggression and impulsivity in healthy adults.

    Science.gov (United States)

    Cook, Thomas B; Brenner, Lisa A; Cloninger, C Robert; Langenberg, Patricia; Igbide, Ajirioghene; Giegling, Ina; Hartmann, Annette M; Konte, Bettina; Friedl, Marion; Brundin, Lena; Groer, Maureen W; Can, Adem; Rujescu, Dan; Postolache, Teodor T

    2015-01-01

    Latent chronic infection with Toxoplasma gondii (T. gondii), a common neurotropic pathogen, has been previously linked with suicidal self-directed violence (SSDV). We sought to determine if latent infection with T. gondii is associated with trait aggression and impulsivity, intermediate phenotypes for suicidal behavior, in psychiatrically healthy adults. Traits of aggression and impulsivity were analyzed in relationship to IgG antibody seropositivity for T. gondii and two other latent neurotropic infections, herpes simplex virus 1 (HSV1) and cytomegalovirus (CMV). One thousand community-residing adults residing in the Munich metropolitan area with no Axis I or II conditions by SCID for DSM-IV (510 men, 490 women, mean age 53.6 ± 15.8, range 20-74). Plasma samples were tested for IgG antibodies to T. gondii, HSV-1 and CMV by ELISA. Self-reported ratings of trait aggression scores (Questionnaire for Measuring Factors of Aggression [FAF]) and trait impulsivity (Sensation-Seeking Scale-V [SSS-V]) were analyzed using linear multivariate methods. T. gondii IgG seropositivity was significantly associated with higher trait reactive aggression scores among women (p impulsive sensation-seeking (SSS-V Disinhibition) among younger men (p impulsivity, personality traits considered as endophenotypes for SSDV, are associated with latent T. gondii infection in a gender and age-specific manner, and could be further investigated as prognostic and treatment targets in T. gondii-positive individuals at risk for SSDV. Published by Elsevier Ltd.

  16. Dual Sarcocystis neurona and Toxoplasma gondii infection in a northern sea otter from Washington state, USA

    Science.gov (United States)

    Lindsay, D.S.; Thomas, N.J.; Rosypal, A.C.; Dubey, J.P.

    2001-01-01

    Dual Sarcocystis neurona and Toxoplasma gondii infection was observed in a Northern sea otter from Washington, USA. The animal was found stranded, convulsed, and died shortly thereafter. Encephalitis caused by both S. neurona and T. gondii was demonstrated in histological sections of brain. Immunohistochemical examination of sections with S. neurona specific antisera demonstrated developmental stages that divided by endopolygeny and produced numerous merozoites. PCR of brain tissue from the sea otter using primer pairs JNB33/JNB54 resulted in amplification of a 1100 bp product. This PCR product was cut in to 884 and 216 bp products by Dra I but was not cut by Hinf I indicating that it was S. neurona [J. Parasitol. 85 (1999) 221]. No PCR product was detected in the brain of a sea otter which had no lesions of encephalitis. Examination of brain sections using T. gondii specific antisera demonstrated tachyzoites and tissue cysts of T. gondii. The lesions induced by T. gondii suggested that the sea otter was suffering from reactivated toxoplasmosis. T. gondii was isolated in mice inoculated with brain tissue. A cat that was fed infected mouse brain tissue excreted T. gondii oocysts which were infective for mice. This is apparently the first report of dual S. neurona and T. gondii in a marine mammal.

  17. Seroepidemiology of Toxoplasma gondii infection in bats from São Paulo city, Brazil

    Science.gov (United States)

    Toxoplasmosis is a parasitic zoonosis caused by Toxoplasma gondii with a worldwide distribution. The aim of this study was to determine the frequency of IgG anti-T. gondii antibodies in bats from São Paulo city, Brazil. A total of 616 serum samples were collected from 22 species of bats. Anti-T. gon...

  18. SAG2 locus genotyping of Toxoplasma gondii in meat products of ...

    African Journals Online (AJOL)

    Toxoplasmosis is an infection caused by Toxoplasma gondii, an intracellular obligate parasite. Its transmission is usually attributed to ingestion of undercooked or raw meat. The aim of this study was the detection and genotyping of T. gondii in meat products using the molecular method in East Azerbaijan. DNA was ...

  19. Toxoplasma gondii IgG antibodies in HIV/AIDS patients attending ...

    African Journals Online (AJOL)

    Background: Toxoplasmosis is caused by Toxoplasma gondii, a parasite that gradually evolved to be the most opportunistic parasite that complicates the course of HIV/AIDS in developing countries. Aim: This study was undertaken to investigate the presence of Toxoplasma gondii IgG antibodies in HIVinfected patients ...

  20. Prevalence of Toxoplasma gondii IgG and IgM and associated risk ...

    African Journals Online (AJOL)

    Prevalence of Toxoplasma gondii IgG and IgM and associated risk factors among HIV-positive and HIV-negative patients in Vhembe district of South Africa. ... shown a high prevalence of T. gondii (IgG) among patients attending different HIV clinics in the Vhembe district with no current infections among pregnant women.

  1. Transplacental transmission of Toxoplasma gondii in minipigs infected with strains of different virulence

    DEFF Research Database (Denmark)

    Jungersen, Gregers; Bille-Hansen, Vivi; Jensen, Lene Bai

    2001-01-01

    Infections with the Zoonotic protozoan Toxoplasma gondii during pregnancy can result in severe fetal infections. To investigate the use of pigs as animal models for congenital toxoplasmosis, tachyzoites of 5 T. gondii strains, with low to intermediate virulence in mice, were intravenously...... animal models fur studies of transplacental transmission and pathogenesis of congenital toxoplasmosis....

  2. Toxoplasma gondii seroprevalence in free-ranging moose (Alces alces) hunted for human consumption in Estonia

    DEFF Research Database (Denmark)

    Remes, Noora; Kärssin, Age; Must, Kärt

    2018-01-01

    In Estonia, northeastern Europe, antibodies against Toxoplasma gondii are common in many host species, including wildlife, domestic animals, and humans. Our nationwide study aimed to estimate T. gondii seroprevalence and its geographical distribution, and to evaluate plausible risk factors for se...

  3. Seroprevalence and isolation of viable Toxoplasma gondii from raptors in the southeastern USA

    Science.gov (United States)

    Raptors are good indicators of the prevalence of Toxoplasma gondii in the environment because they prey on small mammals and birds. These prey species are a major source of infection in felids, which shed the environmentally resistant oocysts. We assessed T. gondii infection in 281 opportunistically...

  4. Isolation of Viable Toxoplasma gondii from Tissues and Feces of Cats from Addis Ababa, Ethiopia

    Science.gov (United States)

    Cats are important in the epidemiology of Toxoplasma gondii because they are the only hosts that excrete environmentally resistant oocysts in feces. In the present study, hearts, serum, and feces from 36 feral cats from Addis Ababa area, Ethiopia were examined for T. gondii infection. Antibodies to ...

  5. Toxoplasma gondii exposure and Parkinson's disease: a case–control study

    Science.gov (United States)

    Alvarado-Esquivel, Cosme; Méndez-Hernández, Edna Madai; Salas-Pacheco, José Manuel; Ruano-Calderón, Luis Ángel; Hernández-Tinoco, Jesús; Arias-Carrión, Oscar; Sánchez-Anguiano, Luis Francisco; Castellanos-Juárez, Francisco Xavier; Sandoval-Carrillo, Ada Agustina; Liesenfeld, Oliver; Ramos-Nevárez, Agar

    2017-01-01

    Objectives To determine the association between Toxoplasma gondii infection and Parkinson's disease and to investigate whether T. gondii seropositivity is associated with the general characteristics of patients with Parkinson's disease. Design Case–control study. Setting Cases and controls were enrolled in Durango City, Mexico. Participants 65 patients with Parkinson's disease and 195 age- and gender-matched control subjects without Parkinson's disease. Primary and secondary outcome measures Serum samples of participants were analysed for anti-T. gondii IgG and IgM antibodies by commercially available enzyme-linked immunoassays. Prevalence of T. gondii DNA was determined in seropositive subjects using PCR. The association between clinical data and infection was examined by bivariate analysis. Results Anti-T. gondii IgG antibodies were found in 6/65 cases (9.2%) and in 21/195 controls (10.8%) (OR 0.84; 95% CI 0.32 to 2.18; p=0.81). The frequency of high (>150 IU/mL) antibody levels was similar among cases and controls (p=0.34). None of the anti-T. gondii IgG positive cases and four of the anti-T. gondii IgG positive controls had anti-T. gondii IgM antibodies (p=0.54). The prevalence of T. gondii DNA was comparable in seropositive cases and controls (16.7% and 25%, respectively; p=1.0). Seroprevalence of T. gondii infection was associated with a young age onset of disease (p=0.03), high Unified Parkinson Disease Rating Scale scores (p=0.04) and depression (p=0.02). Seropositivity to T. gondii infection was lower in patients treated with pramipexole than in patients without this treatment (p=0.01). However, none of the associations remained significant after Bonferroni correction. Conclusions The results do not support an association between T. gondii infection and Parkinson's disease. However, T. gondii infection might have an influence on certain symptoms of Parkinson's disease. Further research to elucidate the role of T. gondii exposure on Parkinson's disease

  6. Prevalence of Toxoplasma gondii and Toxocara canis among Patients with Uveitis.

    Science.gov (United States)

    Lim, Su Jin; Lee, Sang Eun; Kim, Sun Hyun; Hong, Sung-Hee; You, Yong Sung; Kwon, Oh Woong; Kim, Hyeun Seung

    2015-04-01

    To investigate the prevalence of Toxoplasma gondii and Toxocara canis in patients with uveitis. Patients with uveitis were examined. Serum antibodies to T. gondii and T. canis were tested by using the enzyme-linked immunosorbent assay. Polymerase chain reaction (PCR) was done using blood and aqueous humor (AH). Ninety-eight patients were enrolled. Mean age was 43.5 ± 13.2 years. Six patients were seropositive for T. gondii with the following pattern: anterior uveitis, 1; posterior uveitis with retinitis, 2; pan uveitis, 2. One patient had a positive PCR result for T. gondii in AH, who showed panuveitis. Twenty-three patients were positive to serum IgG for T. canis with the following clinical manifestation: granuloma, 6; pigmented scar, 3; vitritis, 6--but none were PCR positive. T. gondii and T. canis are still important causes of uveitis. Ocular toxocariasis is not an uncommon cause of uveitis, even in adults.

  7. Cyclophilin D deficiency rescues Aβ-impaired PKA/CREB signaling and alleviates synaptic degeneration.

    Science.gov (United States)

    Du, Heng; Guo, Lan; Wu, Xiaoping; Sosunov, Alexander A; McKhann, Guy M; Chen, John Xi; Yan, Shirley ShiDu

    2014-12-01

    The coexistence of neuronal mitochondrial pathology and synaptic dysfunction is an early pathological feature of Alzheimer's disease (AD). Cyclophilin D (CypD), an integral part of mitochondrial permeability transition pore (mPTP), is involved in amyloid beta (Aβ)-instigated mitochondrial dysfunction. Blockade of CypD prevents Aβ-induced mitochondrial malfunction and the consequent cognitive impairments. Here, we showed the elimination of reactive oxygen species (ROS) by antioxidants probucol or superoxide dismutase (SOD)/catalase blocks Aβ-mediated inactivation of protein kinase A (PKA)/cAMP regulatory-element-binding (CREB) signal transduction pathway and loss of synapse, suggesting the detrimental effects of oxidative stress on neuronal PKA/CREB activity. Notably, neurons lacking CypD significantly attenuate Aβ-induced ROS. Consequently, CypD-deficient neurons are resistant to Aβ-disrupted PKA/CREB signaling by increased PKA activity, phosphorylation of PKA catalytic subunit (PKA C), and CREB. In parallel, lack of CypD protects neurons from Aβ-induced loss of synapses and synaptic dysfunction. Furthermore, compared to the mAPP mice, CypD-deficient mAPP mice reveal less inactivation of PKA-CREB activity and increased synaptic density, attenuate abnormalities in dendritic spine maturation, and improve spontaneous synaptic activity. These findings provide new insights into a mechanism in the crosstalk between the CypD-dependent mitochondrial oxidative stress and signaling cascade, leading to synaptic injury, functioning through the PKA/CREB signal transduction pathway. Copyright © 2013 Elsevier B.V. All rights reserved.

  8. Cyclophilin B Expression Is Associated with In Vitro Radioresistance and Clinical Outcome after Radiotherapy

    Directory of Open Access Journals (Sweden)

    Paul D. Williams

    2011-12-01

    Full Text Available The tools for predicting clinical outcome after radiotherapy are not yet optimal. To improve on this, we applied the COXEN informatics approach to in vitro radiation sensitivity data of transcriptionally profiled human cells and gene expression data from untreated head and neck squamous cell carcinoma (HNSCC and bladder tumors to generate a multigene predictive model that is independent of histologic findings and reports on tumor radiosensitivity. The predictive ability of this 41-gene model was evaluated in patients with HNSCC and was found to stratify clinical outcome after radiotherapy. In contrast, this model was not useful in stratifying similar patients not treated with radiation. This led us to hypothesize that expression of some of the 41 genes contributes to tumor radioresistance and clinical recurrence. Hence, we evaluated the expression the 41 genes as a function of in vitro radioresistance in the NCI-60 cancer cell line panel and found cyclophilin B (PPIB, a peptidylprolyl isomerase and target of cyclosporine A (CsA, had the strongest direct correlation. Functional inhibition of PPIB by small interfering RNA depletion or CsA treatment leads to radiosensitization in cancer cells and reduced cellular DNA repair. Immunohistochemical evaluation of PPIB expression in patients with HNSCC was found to be associated with outcome after radiotherapy. This work demonstrates that a novel 41-gene expression model of radiation sensitivity developed in bladder cancer cell lines and human skin fibroblasts predicts clinical outcome after radiotherapy in head and neck cancer patients and identifies PPIB as a potential target for clinical radiosensitization.

  9. PpiA, a surface PPIase of the cyclophilin family in Lactococcus lactis.

    Directory of Open Access Journals (Sweden)

    Nicolas Trémillon

    Full Text Available BACKGROUND: Protein folding in the envelope is a crucial limiting step of protein export and secretion. In order to better understand this process in Lactococcus lactis, a lactic acid bacterium, genes encoding putative exported folding factors like Peptidyl Prolyl Isomerases (PPIases were searched for in lactococcal genomes. RESULTS: In L. lactis, a new putative membrane PPIase of the cyclophilin subfamily, PpiA, was identified and characterized. ppiA gene was found to be constitutively expressed under normal and stress (heat shock, H(2O(2 conditions. Under normal conditions, PpiA protein was synthesized and released from intact cells by an exogenously added protease, showing that it was exposed at the cell surface. No obvious phenotype could be associated to a ppiA mutant strain under several laboratory conditions including stress conditions, except a very low sensitivity to H(2O(2. Induction of a ppiA copy provided in trans had no effect i on the thermosensitivity of an mutant strain deficient for the lactococcal surface protease HtrA and ii on the secretion and stability on four exported proteins (a highly degraded hybrid protein and three heterologous secreted proteins in an otherwise wild-type strain background. However, a recombinant soluble form of PpiA that had been produced and secreted in L. lactis and purified from a culture supernatant displayed both PPIase and chaperone activities. CONCLUSIONS: Although L. lactis PpiA, a protein produced and exposed at the cell surface under normal conditions, displayed a very moderate role in vivo, it was found, as a recombinant soluble form, to be endowed with folding activities in vitro.

  10. EMMPRIN and its ligand cyclophilin A as novel diagnostic markers in inflammatory cardiomyopathy.

    Science.gov (United States)

    Seizer, Peter; Geisler, Tobias; Bigalke, Boris; Schneider, Martin; Klingel, Karin; Kandolf, Reinhard; Stellos, Konstantinos; Schreieck, Jürgen; Gawaz, Meinrad; May, Andreas E

    2013-03-10

    During inflammatory cardiomyopathy matrix metalloproteinases are crucially involved in cardiac remodeling. The aim of the present study was to investigate whether the "extracellular matrix metalloproteinase inducer" EMMPRIN (CD147) and its ligand Cyclophilin A (CyPA) are upregulated in inflammatory cardiomyopathy and may serve as diagnostic markers. Therefore, a series of 102 human endomyocardial biopsies were analyzed for the expression of EMMPRIN and CyPA and correlated with histological and immunohistological findings. Endomyocardial biopsies were stained for EMMPRIN and CyPA in addition to standard histology (HE, Trichrom) and immunohistological stainings (MHC-II, CD68, CD3). 39 (38.2%) biopsies met the immunohistological criteria of an inflammatory cardiomyopathy. EMMPRIN, which was predominantly expressed on cardiomyocytes, was slightly (but significantly) upregulated in non inflammatory cardiomyopathies compared to normal histopathological findings and highly upregulated in inflammatory cardiomyopathy compared to both non inflammatory cardiomyopathy and normal histopathology. In contrast, CyPA reveals no enhanced expression in non inflammatory cardiomyopathies and a highly enhanced expression in inflammatory cardiomyopathy, where it is closely associated with leucocytes infiltrates. We found a strong correlation between both EMMPRIN and CyPA with the expression of MHC-II molecules (correlation coefficient 0.475 and 0.527, pEMMPRIN and CyPA with CD68 (correlation coefficient 0.393 and 0.387, pEMMPRIN is enhanced in both inflammatory and non inflammatory cardiomyopathies and can serve as a marker of myocardial remodeling. CyPA may represent a novel and specific marker for cardiac inflammation. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  11. Cyclophilin D Promotes Brain Mitochondrial F1FO ATP Synthase Dysfunction in Aging Mice.

    Science.gov (United States)

    Gauba, Esha; Guo, Lan; Du, Heng

    2017-01-01

    Brain aging is the known strongest risk factor for Alzheimer's disease (AD). In recent years, mitochondrial deficits have been proposed to be a common mechanism linking brain aging to AD. Therefore, to elucidate the causative mechanisms of mitochondrial dysfunction in aging brains is of paramount importance for our understanding of the pathogenesis of AD, in particular its sporadic form. Cyclophilin D (CypD) is a specific mitochondrial protein. Recent studies have shown that F1FO ATP synthase oligomycin sensitivity conferring protein (OSCP) is a binding partner of CypD. The interaction of CypD with OSCP modulates F1FO ATP synthase function and mediates mitochondrial permeability transition pore (mPTP) opening. Here, we have found that increased CypD expression, enhanced CypD/OSCP interaction, and selective loss of OSCP are prominent brain mitochondrial changes in aging mice. Along with these changes, brain mitochondria from the aging mice demonstrated decreased F1FO ATP synthase activity and defective F1FO complex coupling. In contrast, CypD deficient mice exhibited substantially mitigated brain mitochondrial F1FO ATP synthase dysfunction with relatively preserved mitochondrial function during aging. Interestingly, the aging-related OSCP loss was also dramatically attenuated by CypD depletion. Therefore, the simplest interpretation of this study is that CypD promotes F1FO ATP synthase dysfunction and the resultant mitochondrial deficits in aging brains. In addition, in view of CypD and F1FO ATP synthase alterations seen in AD brains, the results further suggest that CypD-mediated F1FO ATP synthase deregulation is a shared mechanism linking mitochondrial deficits in brain aging and AD.

  12. Role of cyclophilin B in prolactin signal transduction and nuclear retrotranslocation.

    Science.gov (United States)

    Rycyzyn, M A; Reilly, S C; O'Malley, K; Clevenger, C V

    2000-08-01

    The pleiotropic actions of PRL are necessary for mammary growth and differentiation and in vitro lymphoid proliferation. The proximal action of this ligand is mediated by its cell surface receptor via associated networks. PRL action, however, is also associated with the internalization and translocation of this hormone into the nucleus. To delineate the mechanism of this retrotranslocation, a yeast two-hybrid screen was performed and revealed an interaction between PRL and cyclophilin B (CypB). CypB is a peptidyl prolyl isomerase (PPI) found in the endoplasmic reticulum, extracellular space, and nucleus. The interaction between CypB and PRL was subsequently confirmed in vitro and in vivo through the use of recombinant proteins and coimmunoprecipitation studies. The exogenous addition of CypB potentiated the 3H-thymidine incorporation of PRL-dependent cell lines up to 18-fold. CypB by itself was nonmitogenic and did not potentiate the action of GH or other interleukins. CypB did not alter the affinity of the PRL receptor (PRLr) for its ligand, or increase the phosphorylation of PRLr-associated Jak2 or Stat5a. The potentiation of PRL-action by CypB, however, was accompanied by a dramatic increase in the nuclear retrotranslocation of PRL. A CypB mutant, termed CypB-NT, was generated that lacked the wild-type N-terminal nuclear localization sequence. Although CypB-NT demonstrated levels of PRL binding and PPI activity equivalent to wild-type CypB, it was incapable of mediating the nuclear retrotranslocation of PRL or enhancing PRL-driven proliferation. These studies reveal CypB as an important chaperone facilitating the nuclear retrotransport and action of the lactogenic hormones.

  13. Distribution of cyclophilin B-binding sites in the subsets of human peripheral blood lymphocytes.

    Science.gov (United States)

    Denys, A; Allain, F; Foxwell, B; Spik, G

    1997-08-01

    Cyclophilin B (CyPB) is a cyclosporin A (CsA)-binding protein, mainly associated with the secretory pathway and released in biological fluids. We have recently demonstrated that both free CyPB and CyPB-CsA complex specifically bind to peripheral blood T lymphocytes and are internalized. These results suggest that CyPB might promote the targeting of the drug into sensitive cells. Peripheral blood lymphocytes are subdivided in several populations according to their biological functions and sensitivity to CsA. We have investigated the binding of CyPB to these different subsets using a CyPB derivatized by fluorescein through its single cysteine which retains its binding properties. We have confirmed that only T cells were involved in the interaction with CyPB. The ligand binding was found to be heterogeneously distributed on the different T-cell subsets and surface-bound CyPB was mainly associated with the CD4-positive cells. No significant difference was noted between the CD45RA and CD45RO subsets, demonstrating that CyPB-binding sites were equally distributed between native and memory T cells. CD3 stimulation of T lymphocytes led to a decrease in the CyPB-binding capacity, that may be explained by a down-regulation of the CyPB-receptor expression upon T-cell activation. Finally, we demonstrated that CyPB-receptor-positive cells, isolated on CyPB sulphydryl-coupled affinity matrices, are more sensitive to CyPB-complexed CsA than mixed peripheral blood lymphocytes, suggesting that CyPB potentiates CsA activity through the binding of the complex. Taken together, our results demonstrate that CyPB-binding sites are mainly associated with resting cells of the helper T lymphocyte, and that CyPB might modulate the distribution of CsA through the drug targeting to sensitive cells.

  14. Cyclophilins contribute to Stat3 signaling and survival of multiple myeloma cells.

    Science.gov (United States)

    Bauer, K; Kretzschmar, A K; Cvijic, H; Blumert, C; Löffler, D; Brocke-Heidrich, K; Schiene-Fischer, C; Fischer, G; Sinz, A; Clevenger, C V; Horn, F

    2009-08-06

    Signal transducer and activator of transcription 3 (Stat3) is the major mediator of interleukin-6 (IL-6) family cytokines. In addition, Stat3 is known to be involved in the pathophysiology of many malignancies. Here, we show that the cis-trans peptidyl-prolyl isomerase cyclophilin (Cyp) B specifically interacts with Stat3, whereas the highly related CypA does not. CypB knockdown inhibited the IL-6-induced transactivation potential but not the tyrosine phosphorylation of Stat3. Binding of CypB to Stat3 target promoters and alteration of the intranuclear localization of Stat3 on CypB depletion suggested a nuclear function of Stat3/CypB interaction. By contrast, CypA knockdown inhibited Stat3 IL-6-induced tyrosine phosphorylation and nuclear translocation. The Cyp inhibitor cyclosporine A (CsA) caused similar effects. However, Stat1 activation in response to IL-6 or interferon-gamma was not affected by Cyp silencing or CsA treatment. As a result, Cyp knockdown shifted IL-6 signaling to a Stat1-dominated pathway. Furthermore, Cyp depletion or treatment with CsA induced apoptosis in IL-6-dependent multiple myeloma cells, whereas an IL-6-independent line was not affected. Thus, Cyps support the anti-apoptotic action of Stat3. Taken together, CypA and CypB both play pivotal roles, yet at different signaling levels, for Stat3 activation and function. These data also suggest a novel mechanism of CsA action.

  15. Receptor-mediated transcytosis of cyclophilin B through the blood-brain barrier.

    Science.gov (United States)

    Carpentier, M; Descamps, L; Allain, F; Denys, A; Durieux, S; Fenart, L; Kieda, C; Cecchelli, R; Spik, G

    1999-07-01

    Cyclophilin B (CyPB) is a cyclosporin A (CsA)-binding protein mainly located in intracellular vesicles and secreted in biological fluids. In previous works, we demonstrated that CyPB interacts with T lymphocytes and enhances in vitro cellular incorporation and activity of CsA. In addition to its immunosuppressive activity, CsA is able to promote regeneration of damaged peripheral nerves. However, the crossing of the drug from plasma to neural tissue is restricted by the relative impermeability of the blood-brain barrier. To know whether CyPB might also participate in the delivery of CsA into the brain, we have analyzed the interactions of CyPB with brain capillary endothelial cells. First, we demonstrated that CyPB binds to two types of binding sites present at the surface of capillary endothelial cells from various species of tissues. The first type of binding sites (K(D) = 300 nM; number of sites = 3 x 10(6)) is related to interactions with negatively charged compounds such as proteoglycans. The second type of binding sites, approximately 50,000 per cell, exhibits a higher affinity for CyPB (K(D) = 15 nM) and is involved in an endocytosis process, indicating it might correspond to a functional receptor. Finally, the use of an in vitro model of blood-brain barrier allowed us to demonstrate that CyPB is transcytosed by a receptor-mediated pathway (flux = 16.5 fmol/cm2/h). In these conditions, CyPB did not significantly modify the passage of CsA, indicating that it is unlikely to provide a pathway for CsA brain delivery.

  16. Different mechanisms of hepatitis C virus RNA polymerase activation by cyclophilin A and B in vitro.

    Science.gov (United States)

    Weng, Leiyun; Tian, Xiao; Gao, Yayi; Watashi, Koichi; Shimotohno, Kunitada; Wakita, Takaji; Kohara, Michinori; Toyoda, Tetsuya

    2012-12-01

    Cyclophilins (CyPs) are cellular proteins that are essential to hepatitis C virus (HCV) replication. Since cyclosporine A was discovered to inhibit HCV infection, the CyP pathway contributing to HCV replication is a potential attractive stratagem for controlling HCV infection. Among them, CyPA is accepted to interact with HCV nonstructural protein (NS) 5A, although interaction of CyPB and NS5B, an RNA-dependent RNA polymerase (RdRp), was proposed first. CyPA, CyPB, and HCV RdRp were expressed in bacteria and purified using combination column chromatography. HCV RdRp activity was analyzed in vitro with purified CyPA and CyPB. CyPA at a high concentration (50× higher than that of RdRp) but not at low concentration activated HCV RdRp. CyPB had an allosteric effect on genotype 1b RdRp activation. CyPB showed genotype specificity and activated genotype 1b and J6CF (2a) RdRps but not genotype 1a or JFH1 (2a) RdRps. CyPA activated RdRps of genotypes 1a, 1b, and 2a. CyPB may also support HCV genotype 1b replication within the infected cells, although its knockdown effect on HCV 1b replicon activity was controversial in earlier reports. CyPA activated HCV RdRp at the early stages of transcription, including template RNA binding. CyPB also activated genotype 1b RdRp. However, their activation mechanisms are different. These data suggest that both CyPA and CyPB are excellent targets for the treatment of HCV 1b, which shows the greatest resistance to interferon and ribavirin combination therapy. Copyright © 2012 Elsevier B.V. All rights reserved.

  17. Enhancing the effect of secreted cyclophilin B on immunosuppressive activity of cyclosporine.

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    Denys, A; Allain, F; Masy, E; Dessaint, J P; Spik, G

    1998-04-27

    Cyclophilin B (CyPB) is a cyclosporine (CsA)-binding protein, located within intracellular vesicles and secreted in biological fluids. In previous works, we reported that CyPB specifically interacts with the T-cell membrane and potentiates the ability of CsA to inhibit CD3-induced proliferation of T lymphocytes. CyPB levels were measured in plasma from healthy donors and transplant patients. The role of extracellular CyPB on the distribution and activity of CsA was investigated first by studies on the uptake of free and CyPB-complexed drug by blood cells, and second by studies on the inhibitory effects of these two compounds on the CD3-induced proliferation of peripheral blood mononuclear cells. A significant increase in plasma CyPB level was observed for CsA-treated patients (13+/-6.4 nM, n=42) in comparison with untreated donors (4.3+/-2.1 nM, n=34). In vitro, extracellular CyPB dose dependently modified CsA distribution between plasma, erythrocyte, and lymphocyte contents, by both retaining the complexed drug extracellularly and promoting its specific accumulation within peripheral blood mononuclear cells. Moreover, the enhanced ability of CyPB-complexed CsA to suppress CD3-induced T-cell proliferation was preserved in the presence of other blood cells, implying specific targeting of the drug to sensitive cells. Furthermore, although a large interindividual variability of sensitivity to the drug was confirmed for 18 individuals, we found that CyPB potentiated the activity of CsA in restoring a high sensitivity to the immunosuppressant. These results suggest that plasma CyPB may contribute to the acceptance and the good maintenance of organ transplantation by enhancing the immunosuppressive activity of CsA through a receptor-mediated incorporation of CyPB-complexed CsA within peripheral blood lymphocytes.

  18. The human TRPV6 channel protein is associated with cyclophilin B in human placenta.

    Science.gov (United States)

    Stumpf, Tobias; Zhang, Qi; Hirnet, Daniela; Lewandrowski, Urs; Sickmann, Albert; Wissenbach, Ulrich; Dörr, Janka; Lohr, Christian; Deitmer, Joachim W; Fecher-Trost, Claudia

    2008-06-27

    Transcellular calcium transport in the kidney, pancreas, small intestine, and placenta is partly mediated by transient receptor potential (TRP) channels. The highly selective TRPV6 calcium channel protein is most likely important for the calcium transfer in different specialized epithelial cells. In the human placenta the protein is expressed in trophoblast tissue, where it is implicated in the transepithelial calcium transfer from mother to the fetus. We enriched the TRPV6 channel protein endogenously expressed in placenta together with annexin A2 and cyclophilin B (CypB), which is a member of the huge immunophilin family. In the human placenta TRPV6 and CypB are mainly located intracellularly in the syncytiotrophoblast layer, but a small amount of the mature glycosylated TRPV6 channel protein and CypB is also expressed in microvilli apical membranes, the fetomaternal barrier. To understand the role of CypB on the TRPV6 channel function, we evaluated the effect of CypB co-expression on TRPV6-mediated calcium uptake into Xenopus laevis oocytes expressing TRPV6. A significant increase of TRPV6-mediated calcium uptake was observed after CypB/TRPV6 co-expression. This stimulatory effect of CypB was reversed by the immunosuppressive drug cyclosporin A, which inhibits the enzymatic activity of CypB. Cyclosporin A had no significant effect on TRPV6 and CypB protein expression levels in the oocytes. In summary, our results establish CypB as a new TRPV6 accessory protein with potential involvement in TRPV6 channel activation through its peptidyl-prolyl cis/trans isomerase activity.

  19. The SARS-coronavirus-host interactome: identification of cyclophilins as target for pan-coronavirus inhibitors.

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    Susanne Pfefferle

    2011-10-01

    Full Text Available Coronaviruses (CoVs are important human and animal pathogens that induce fatal respiratory, gastrointestinal and neurological disease. The outbreak of the severe acute respiratory syndrome (SARS in 2002/2003 has demonstrated human vulnerability to (Coronavirus CoV epidemics. Neither vaccines nor therapeutics are available against human and animal CoVs. Knowledge of host cell proteins that take part in pivotal virus-host interactions could define broad-spectrum antiviral targets. In this study, we used a systems biology approach employing a genome-wide yeast-two hybrid interaction screen to identify immunopilins (PPIA, PPIB, PPIH, PPIG, FKBP1A, FKBP1B as interaction partners of the CoV non-structural protein 1 (Nsp1. These molecules modulate the Calcineurin/NFAT pathway that plays an important role in immune cell activation. Overexpression of NSP1 and infection with live SARS-CoV strongly increased signalling through the Calcineurin/NFAT pathway and enhanced the induction of interleukin 2, compatible with late-stage immunopathogenicity and long-term cytokine dysregulation as observed in severe SARS cases. Conversely, inhibition of cyclophilins by cyclosporine A (CspA blocked the replication of CoVs of all genera, including SARS-CoV, human CoV-229E and -NL-63, feline CoV, as well as avian infectious bronchitis virus. Non-immunosuppressive derivatives of CspA might serve as broad-range CoV inhibitors applicable against emerging CoVs as well as ubiquitous pathogens of humans and livestock.

  20. Toxoplasma gondii in Blood Donors: A Study in Boyer-Ahmad County, Southwest Iran

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    Abdolali Moshfe

    2018-01-01

    Full Text Available Toxoplasma gondii is an important foodborne protozoan that can be transmitted through infected blood containing tachyzoite form of the parasite. The current study aimed to evaluate the prevalence of T. gondii infection and related risk factors among healthy blood donors in Boyer-Ahmad County, southwest Iran. Blood samples were taken from 285 healthy blood donors who voluntarily agreed to participate in this study. Sera and buffy coat were isolated from the blood samples for serological and molecular evaluations. The sera were tested for anti-T. gondii antibodies (both IgG and IgM, using a commercial ELISA kit. The buffy coat of seropositive cases was evaluated for detection of T. gondii DNA by PCR. Moreover, a structured questionnaire, containing socioepidemiological data and possible risk factors, was filled out by each participant during sample collection. Anti-T. gondii antibodies were detected in sera of 48/285 (16.8% participants. Only two of the subjects (0.7% were seropositive for both IgG and IgM antibodies. T. gondii DNA was not detected in buffy coat of any of the seropositive cases. Risk factors such as contact with soil (OR, 9.7; 95% CI, 4.9–19.4 and consumption of semicooked meat (OR, 2.5; 95% CI, 1.2–5.03 were statistically associated with seropositivity to T. gondii. The seroprevalence rate of T. gondii antibodies in the blood donors of Boyer-Ahmad County was not high in comparison with other regions in Iran. In this study, consumption of undercooked meats, job, and contact with soil were independent risk factors associated with T. gondii infection, which can be considered as potential sources of T. gondii infection.

  1. Prevalence and genetic characterization of Toxoplasma gondii in donkeys in northeastern China.

    Science.gov (United States)

    Zhang, Xiao-Xuan; Shi, Wei; Zhang, Nian-Zhang; Shi, Kun; Li, Jian-Ming; Xu, Peng; Zhao, Quan; Du, Rui

    2017-10-01

    Toxolasma gondii is one of the most important obligate intracellular protozoan parasites. Recently, toxoplasmosis is of increasing concerns because T. gondii not only has a worldwide distribution but also can infect virtually all warm-blooded hosts including donkeys. However, limited information is available concerning the genetic characterization of T. gondii in donkeys in northeastern China. In this study, a total of 302 brain tissue samples collected from donkeys from Jilin (n=108) and Liaoning (n=194) provinces, northeastern China, were examined for T. gondii infection by semi-nested PCR of B1 gene. The positive samples were genotyped at 11 loci (i.e., SAG1, alternative SAG2, 5'-and 3'-SAG2, SAG3, L358, BTUB, c22-8, GRA6, c29-2, PK1 and Apico) using polymerase chain reaction-restriction fragment length polymorphism technology. Of 302 brain tissue samples, 19 (6.29%) were PCR-positive for T. gondii. The prevalence rates of T. gondii were 6.48% (7/108) and 6.19% (12/194) in Jilin Province and Liaoning Province, respectively. The prevalence of T. gondii in different season groups varied from 5.56% to 7.41%. The prevalence of T. gondii in "Cage-free" donkeys was higher than that in "Captive" donkeys. Of the 19 positive samples, only two isolates were successfully genotyped at all loci, three were genotyped at 9 loci. In total, four samples belong to ToxoDB genotype #9, one belong to ToxoDB genotype #10. This is firstly characterized the T. gondii isolates from donkeys in northeastern China. The results of the present study will improve the information of the distribution of T. gondii genotypes in China. Copyright © 2017. Published by Elsevier B.V.

  2. Toxoplasma gondii: infection natural congenital in cattle and an experimental inoculation of gestating cows with oocysts.

    Science.gov (United States)

    Costa, Gustavo Henrique Nogueira; da Costa, Alvimar José; Lopes, Welber Daniel Zanetti; Bresciani, Katia Denise Saraiva; dos Santos, Thais Rabelo; Esper, César Roberto; Santana, Aureo Evangelista

    2011-01-01

    Two studies, of a natural infection and an experimental infection, were performed in order to study congenital transmission of Toxoplasma gondii in cattle. In the first study, 50 fetuses were harvested from gestating cows that were eutanasied at a municipal slaughterhouse in Jaboticabal, São Paulo state, Brazil. In the second study, 11 gestating cows were divided into four groups for inoculation with T. gondii: GI consisted of three cows inoculated with 1.0 × 10(5) oocysts during their first trimester of gestation; GII consisted of three cows inoculated with 1.0 × 10(5) oocysts during their second trimester of gestation; GIII consisted of three cows inoculated with 1.0 × 10(5) oocysts during their last trimester of gestation; and GIV consisted of two control cows, one during its first and the other during its second trimester of gestation. In both studies, the presence of T. gondii was confirmed both indirectly by immunofluorescence assay (IFAT). In the natural infection experiment, 18% (9/50) of the gestating cows were confirmed to have specific antibodies (IFAT--1:64) against T. gondii. The bioassay was able to diagnose the presence of T. gondii in the tissue samples from three calves. In the second experiment, the nine cows from groups I, II and III presented with specific antibodies (IFAT) against T. gondii. In contrast, T. gondii could not be detected by IFAT, histopathological examination or the bioassay in any of the nine calves born to cows experimentally infected with T. gondii oocysts. Based on the results from both studies, we conclude that congenital infection of T. gondii in cattle, while infrequent, does occur naturally. The pathogenicity of the strain of T. gondii may influence the likelihood of this route of transmission. Copyright © 2010 Elsevier Inc. All rights reserved.

  3. Detection and genotyping of Toxoplasma gondii DNA in the blood and milk of naturally infected donkeys (Equus asinus).

    Science.gov (United States)

    Mancianti, Francesca; Nardoni, Simona; Papini, Roberto; Mugnaini, Linda; Martini, Mina; Altomonte, Iolanda; Salari, Federica; D'Ascenzi, Carlo; Dubey, Jitender P

    2014-04-03

    Toxoplasma gondii is a worldwide zoonotic protozoan. Consumption of raw milk from infected animals is considered a risk factor for acquiring toxoplasmosis in humans. Recently, donkey milk has been indicated for therapeutic and nutritional purposes and T. gondii infection is common in donkeys. The purpose of the present paper was to detect the presence of parasite DNA in milk of T. gondii positive donkeys. Antibodies to T. gondii were found in 11 out of 44 healthy lactating donkeys by IFAT. T. gondii DNA was detected by PCR in blood of 6 and milk of 3 seropositive jennies. Results of limited RFLP-PCR genotyping indicated the presence of T. gondii genotype II or III, commonly found in Europe. The occurrence of T. gondii DNA in milk suggests that the consumption of raw milk from seropositive donkeys could be a potential source of human infection.

  4. Expression, purification, crystallization and preliminary X-ray diffraction analysis of Arabidopsis thaliana cyclophilin 38 (AtCyp38)

    International Nuclear Information System (INIS)

    Vasudevan, Dileep; Gopalan, Gayathri; He, Zengyong; Luan, Sheng; Swaminathan, Kunchithapadam

    2005-01-01

    Crystallization of Arabidopsis thaliana cyclophilin 38. The crystal diffracts X-rays to 2.5 Å resolution. AtCyp38 is one of the highly divergent multidomain cyclophilins from Arabidopsis thaliana. A recombinant form of AtCyp38 (residues 83–437) was expressed in Escherichia coli and purified to homogeneity. The protein was crystallized using the vapour-batch technique with PEG 6000 and t-butanol as precipitants. Crystals of recombinant AtCyp38 diffracted X-rays to better than 2.5 Å resolution at 95 K using a synchrotron-radiation source. The crystal belongs to the C-centred orthorhombic space group C222 1 , with unit-cell parameters a = 58.2, b = 95.9, c = 167.5 Å, and contains one molecule in the asymmetric unit. The selenomethionine derivative of the AtCyp38 protein was overexpressed, purified and crystallized in the same space group and data were collected to 3.5 Å at the NSLS synchrotron. The structure is being solved by the MAD method

  5. Cellular growth kinetics distinguish a cyclophilin inhibitor from an HSP90 inhibitor as a selective inhibitor of hepatitis C virus.

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    Rudolf K F Beran

    Full Text Available During antiviral drug discovery, it is critical to distinguish molecules that selectively interrupt viral replication from those that reduce virus replication by adversely affecting host cell viability. In this report we investigate the selectivity of inhibitors of the host chaperone proteins cyclophilin A (CypA and heat-shock protein 90 (HSP90 which have each been reported to inhibit replication of hepatitis C virus (HCV. By comparing the toxicity of the HSP90 inhibitor, 17-(Allylamino-17-demethoxygeldanamycin (17-AAG to two known cytostatic compounds, colchicine and gemcitabine, we provide evidence that 17-AAG exerts its antiviral effects indirectly through slowing cell growth. In contrast, a cyclophilin inhibitor, cyclosporin A (CsA, exhibited selective antiviral activity without slowing cell proliferation. Furthermore, we observed that 17-AAG had little antiviral effect in a non-dividing cell-culture model of HCV replication, while CsA reduced HCV titer by more than two orders of magnitude in the same model. The assays we describe here are useful for discriminating selective antivirals from compounds that indirectly affect virus replication by reducing host cell viability or slowing cell growth.

  6. Cyclophilin B protects SH-SY5Y human neuroblastoma cells against MPP(+)-induced neurotoxicity via JNK pathway.

    Science.gov (United States)

    Oh, Yoojung; Jeong, Kwon; Kim, Kiyoon; Lee, Young-Seok; Jeong, Suyun; Kim, Sung Soo; Yoon, Kyung-Sik; Ha, Joohun; Kang, Insug; Choe, Wonchae

    2016-09-23

    Parkinson's disease (PD) is the second most common neurodegenerative disorder of aging. PD involves a progressive loss of dopaminergic neurons in the substantia nigra pars compacta. 1-Methyl-4-phenyl-1, 2, 3, 6-tetrahydropyidine (MPTP) and its toxic metabolite 1-methyl-4-phenylpyridinium ion (MPP+) inhibit the complex I of the mitochondrial electron transport chain, and have been widely used to construct PD models. Cyclophilin B (CypB) is an endoplasmic reticulum protein that binds to cyclosporine A as a cyclophilin family member. CypB has peptidyl-prolyl cis-trans isomerase (PPIase) activity. We investigated the protective effects of overexpressed CypB on MPP+-induced neurocytotoxicity in SH-SY5Y human neuroblastoma cells. Overexpressed CypB decreased MPP(+)-induced oxidative stress through the modulation of antioxidant enzymes including manganese superoxide dismutase and catalase, and prevented neurocytotoxicity via mitogen-activated protein kinase, especially the c-Jun N-terminal kinase pathway. In addition, CypB inhibited the activation of MPP(+)-induced the pro-apoptotic molecules poly (ADP-ribose) polymerase, Bax, and Bcl-2, and attenuated MPP(+)-induced mitochondrial dysfunction. The data suggest that overexpressed CypB protects neuronal cells from MPP+-induced dopaminergic neuronal cell death. Copyright © 2016 Elsevier Inc. All rights reserved.

  7. Cloning, purification, crystallization and preliminary X-ray crystallographic analysis of a cyclophilin A-like protein from Piriformospora indica

    International Nuclear Information System (INIS)

    Bhatt, Harshesh; Trivedi, Dipesh Kumar; Pal, Ravi Kant; Johri, Atul Kumar; Tuteja, Narendra; Bhavesh, Neel Sarovar

    2012-01-01

    Cyclophin A like protein from Piriformospora indica involved in salt-stress tolerance was cloned, overexpressed, purified and crystallized. The crystals obtained diffracted X-rays to 2 Å resolution and belonged to space group C222 1 . Cyclophilins are widely distributed both in eukaryotes and prokaryotes and have a primary role as peptidyl-prolyl cis–trans isomerases (PPIases). This study focuses on the cloning, expression, purification and crystallization of a salinity-stress-induced cyclophilin A (CypA) homologue from the symbiotic fungus Piriformospora indica. Crystallization experiments in the presence of 56 mM sodium phosphate monobasic monohydrate, 1.34 M potassium phosphate dibasic pH 8.2 yielded crystals that were suitable for X-ray diffraction analysis. The crystals belonged to the orthorhombic space group C222 1 , with unit-cell parameters a = 121.15, b = 144.12, c = 110.63 Å. The crystals diffracted to a resolution limit of 2.0 Å. Analysis of the diffraction data indicated the presence of three molecules of the protein per asymmetric unit (V M = 4.48 Å 3 Da −1 , 72.6% solvent content)

  8. Sequence Variation in Toxoplasma gondii rop17 Gene among Strains from Different Hosts and Geographical Locations

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    Nian-Zhang Zhang

    2014-01-01

    Full Text Available Genetic diversity of T. gondii is a concern of many studies, due to the biological and epidemiological diversity of this parasite. The present study examined sequence variation in rhoptry protein 17 (ROP17 gene among T. gondii isolates from different hosts and geographical regions. The rop17 gene was amplified and sequenced from 10 T. gondii strains, and phylogenetic relationship among these T. gondii strains was reconstructed using maximum parsimony (MP, neighbor-joining (NJ, and maximum likelihood (ML analyses. The partial rop17 gene sequences were 1375 bp in length and A+T contents varied from 49.45% to 50.11% among all examined T. gondii strains. Sequence analysis identified 33 variable nucleotide positions (2.1%, 16 of which were identified as transitions. Phylogeny reconstruction based on rop17 gene data revealed two major clusters which could readily distinguish Type I and Type II strains. Analyses of sequence variations in nucleotides and amino acids among these strains revealed high ratio of nonsynonymous to synonymous polymorphisms (>1, indicating that rop17 shows signs of positive selection. This study demonstrated the existence of slightly high sequence variability in the rop17 gene sequences among T. gondii strains from different hosts and geographical regions, suggesting that rop17 gene may represent a new genetic marker for population genetic studies of T. gondii isolates.

  9. Seroprevalence of Toxoplasma gondii in humans and pigs in North Sulawesi, Indonesia.

    Science.gov (United States)

    Tuda, Josef; Adiani, Sri; Ichikawa-Seki, Madoka; Umeda, Kousuke; Nishikawa, Yoshifumi

    2017-10-01

    Toxoplasma gondii, an intracellular protozoan parasite, is a major public health concern throughout the world. Importantly, toxoplasmosis has several adverse effects, including neurological and ocular diseases. There are currently no data on the prevalence of T. gondii infection in humans or animals in North Sulawesi, although Indonesia is known to have a high seroprevalence of this parasite. In this study, the prevalence of T. gondii was determined in samples of humans and pigs from North Sulawesi using the latex agglutination test. In total, 856 human were sampled and 58.5% of whom were positive for T. gondii. Although the antibody prevalence in male and female children aged 0-9years was 40% in both sexes, suggesting that the transmission rate of the parasite to humans is extremely high in this area. However, the overall prevalence of T. gondii in pigs was only 2.3%. Our study indicates a high incidence of T. gondii infection in humans. Therefore, a survey of the prevalence of T. gondii among different infection sources is required to determine the major risk factors for infection in North Sulawesi. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Non-association between anti-Toxoplasma gondii antibodies and ABO blood group system

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    ACF Rodrigues

    2011-01-01

    Full Text Available Toxoplasma gondii infects humans through the gastrointestinal tract (GIT, which elicits humoral immune response with specific antibodies. The expression of the ABO blood group glycoconjugates also occurs in this same system and may influence the human susceptibility of infection by T. gondii. The aim of the present study was to investigate the association between ABO blood group phenotypes and the presence of anti-T. gondii antibodies. Data - including age, results of serology tests for T. gondii infection and ABO blood group phenotypes - were assembled from the medical records of 1,006 pregnant women attended in the Base Hospital of the Medical School of São José do Rio Preto, Brazil, between 2001 and 2004. The chi-square test was used to compare the results with the level of significance set at 5%. Of the studied cases, 64.1% (645/1006 and 35.9% (391/1006 presented respectively positive and negative serology tests for anti-T. gondii antibodies. The mean age of those who tested positive was higher than those with negative serology tests (p = 0.0004. The frequencies of ABO blood group phenotypes were similar in those with and without anti-T. gondii antibodies (p = 0.35. In conclusion, the ABO blood group system is not associated with the presence or absence of anti-T. gondii antibodies.

  11. Prevalence of Toxoplasma gondii infection in household and feral cats in Korea.

    Science.gov (United States)

    Kim, Sung-Eon; Choi, Ran; Kang, Seung-Won; Hyun, Changbaig

    2017-09-01

    This study was designed to investigate the prevalence rate of Toxoplasma gondii ( T. gondii ) infection in household cats in Korea. One hundred household cats and 50 feral cats from nine of the largest cities in Korea were enrolled in this study. The tests performed in this survey was an in-house rapid screen IgG and IgM combo test, faecal PCR test for T. gondii oocysts, and an ELISA immunoassay for IgG antibodies. There were no household cats positive for T. gondii infection detected using the in-house IgG and IgM rapid screen combo test, although 6/50 and 0/50 feral cats were positive in IgG and IgM tests, respectively. This initial finding was confirmed by subsequent ELISA test for IgG antibody and PCR for T. gondii in faeces. Despite the higher prevalence rate of the disease in feral cats in Korea, we did not find any household cats that were either infected or exposed previously to T. gondii in our study population. Our study indicates that there is minimal risk of T. gondii transmission from household cats to human in Korea.

  12. LILRB4 Decrease on uDCs Exacerbate Abnormal Pregnancy Outcomes Following Toxoplasma gondii Infection

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    Shaowei Zhan

    2018-03-01

    Full Text Available Toxoplasma gondii (T. gondii infection in early pregnancy can result in miscarriage, dead fetus, and other abnormalities. The LILRB4 is a central inhibitory receptor in uterine dendritic cells (uDCs that plays essential immune-regulatory roles at the maternal–fetal interface. In this study, T. gondii-infected human primary uDCs and T. gondii-infected LILRB4-/- pregnant mice were utilized. The immune mechanisms underlying the role of LILRB4 on uDCs were explored in the development of abnormal pregnancy outcomes following T. gondii infection in vitro and in vivo. Our results showed that the expression levels of LILRB4 on uDCs from normal pregnant mice were obviously higher than non-pregnant mice, and peaked in mid-gestation. The LILRB4 expression on uDC subsets, especially tolerogenic subsets, from mid-gestation was obviously down-regulated after T. gondii infection and LILRB4 decrease could further regulate the expression of functional molecules (CD80, CD86, and HLA-DR or MHC II on uDCs, contributing to abnormal pregnancy outcomes. Our results will shed light on the molecular immune mechanisms of uDCs in abnormal pregnancy outcomes by T. gondii infection.

  13. Concentration and retention of Toxoplasma gondii surrogates from seawater by red abalone (Haliotis rufescens)

    Science.gov (United States)

    Schott, Kristen C; Krusor, Colin; Tinker, M. Tim; Moore, James G.; Conrad, Patricia A.; Shapiro, Karen

    2016-01-01

    Small marine snails and abalone have been identified as high- and low-risk prey items, respectively, for exposure of threatened southern sea otters to Toxoplasma gondii, a zoonotic parasite that can cause fatal encephalitis in animals and humans. While recent work has characterized snails as paratenic hosts for T. gondii, the ability of abalone to vector the parasite has not been evaluated. To further elucidate why abalone predation may be protective against T. gondii exposure, this study aimed to determine whether: (1) abalone are physiologically capable of acquiring T. gondii; and (2) abalone and snails differ in their ability to concentrate and retain the parasite. Abalone were exposed to T. gondii surrogate microspheres for 24 h, and fecal samples were examined for 2 weeks following exposure. Concentration of surrogates was 2–3 orders of magnitude greater in abalone feces than in the spiked seawater, and excretion of surrogates continued for 14 days post-exposure. These results indicate that, physiologically, abalone and snails can equally vector T. gondii as paratenic hosts. Reduced risk of T. gondii infection in abalone-specializing otters may therefore result from abalone's high nutritional value, which implies otters must consume fewer animals to meet their caloric needs.

  14. Concentration and retention of Toxoplasma gondii surrogates from seawater by red abalone (Haliotis rufescens).

    Science.gov (United States)

    Schott, Kristen C; Krusor, Colin; Tinker, M Tim; Moore, James; Conrad, Patricia A; Shapiro, Karen

    2016-11-01

    Small marine snails and abalone have been identified as high- and low-risk prey items, respectively, for exposure of threatened southern sea otters to Toxoplasma gondii, a zoonotic parasite that can cause fatal encephalitis in animals and humans. While recent work has characterized snails as paratenic hosts for T. gondii, the ability of abalone to vector the parasite has not been evaluated. To further elucidate why abalone predation may be protective against T. gondii exposure, this study aimed to determine whether: (1) abalone are physiologically capable of acquiring T. gondii; and (2) abalone and snails differ in their ability to concentrate and retain the parasite. Abalone were exposed to T. gondii surrogate microspheres for 24 h, and fecal samples were examined for 2 weeks following exposure. Concentration of surrogates was 2-3 orders of magnitude greater in abalone feces than in the spiked seawater, and excretion of surrogates continued for 14 days post-exposure. These results indicate that, physiologically, abalone and snails can equally vector T. gondii as paratenic hosts. Reduced risk of T. gondii infection in abalone-specializing otters may therefore result from abalone's high nutritional value, which implies otters must consume fewer animals to meet their caloric needs.

  15. Spatial epidemiology of Toxoplasma gondii infection in goats in Serbia

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    Vitomir Djokić

    2014-05-01

    Full Text Available A major risk factor for Toxoplasma gondii infection is consumption of undercooked meat. Increasing demand for goat meat is likely to promote the role of this animal for human toxoplasmosis. As there are virtually no data on toxoplasmosis in goats in Serbia, we undertook a cross-sectional serological study, including prediction modelling using geographical information systems (GIS. Sera from 431 goats reared in 143 households/farms throughout Serbia, sampled between January 2010 and September 2011, were examined for T. gondii antibodies by a modified agglutination test. Seroprevalence was 73.3% at the individual level and 84.6% at the farm level. Risk factor analysis showed above two-fold higher risk of infection for goats used for all purposes compared to dairy goats (P = 0.012, almost seven-fold higher risk for goats kept as sole species versus those kept with other animals (P = 0.001 and a two-fold lower risk for goats introduced from outside the farm compared to those raised on the farm (P = 0.027. Moreover, households/farms located in centre-eastern Serbia were found to be less often infected than those in northern Serbia (P = 0.004. The risk factor analysis was fully supported by spatial analysis based on a GIS database containing data on origin, serology, land cover, elevation, meteorology and a spatial prediction map based on kriging analysis, which showed western Serbia as the area most likely for finding goats positive for T. gondii and centre-eastern Serbia as the least likely. In addition, rainfall favoured seropositivity, whereas temperature, humidity and elevation did not.

  16. Toxoplasma gondii Actively Inhibits Neuronal Function in Chronically Infected Mice

    Science.gov (United States)

    Haroon, Fahad; Händel, Ulrike; Angenstein, Frank; Goldschmidt, Jürgen; Kreutzmann, Peter; Lison, Holger; Fischer, Klaus-Dieter; Scheich, Henning; Wetzel, Wolfram; Schlüter, Dirk; Budinger, Eike

    2012-01-01

    Upon infection with the obligate intracellular parasite Toxoplasma gondii, fast replicating tachyzoites infect a broad spectrum of host cells including neurons. Under the pressure of the immune response, tachyzoites convert into slow-replicating bradyzoites, which persist as cysts in neurons. Currently, it is unclear whether T. gondii alters the functional activity of neurons, which may contribute to altered behaviour of T. gondii–infected mice and men. In the present study we demonstrate that upon oral infection with T. gondii cysts, chronically infected BALB/c mice lost over time their natural fear against cat urine which was paralleled by the persistence of the parasite in brain regions affecting behaviour and odor perception. Detailed immunohistochemistry showed that in infected neurons not only parasitic cysts but also the host cell cytoplasm and some axons stained positive for Toxoplasma antigen suggesting that parasitic proteins might directly interfere with neuronal function. In fact, in vitro live cell calcium (Ca2+) imaging studies revealed that tachyzoites actively manipulated Ca2+ signalling upon glutamate stimulation leading either to hyper- or hypo-responsive neurons. Experiments with the endoplasmatic reticulum Ca2+ uptake inhibitor thapsigargin indicate that tachyzoites deplete Ca2+ stores in the endoplasmatic reticulum. Furthermore in vivo studies revealed that the activity-dependent uptake of the potassium analogue thallium was reduced in cyst harbouring neurons indicating their functional impairment. The percentage of non-functional neurons increased over time In conclusion, both bradyzoites and tachyzoites functionally silence infected neurons, which may significantly contribute to the altered behaviour of the host. PMID:22530040

  17. Group 1 innate lymphoid cells in Toxoplasma gondii infection.

    Science.gov (United States)

    Dunay, I R; Diefenbach, A

    2018-02-01

    Innate lymphoid cells (ILCs) are a group of lymphocytes that carry out important functions in immunity to infections and in organ homeostasis at epithelial barrier surfaces. ILCs are innate immune cells that provide an early source of cytokines to initiate immune responses against pathogens. Cytotoxic ILCs (i.e. conventional (c)NK cells) and several subsets of helper-like ILCs are the major branches of the ILC family. Conventional NK cells and group 1 ILCs share several characteristics such as surface receptors and the ability to produce IFN-γ upon activation, but they differ in their developmental paths and in their dependence on specific transcription factors. Infection of mice with the intracellular parasite Toxoplasma gondii is followed by a strong Th1-mediated immune response. Previous studies indicate that NK1.1 + cells contribute to the production of IFN-γ and TNF and cytotoxicity during acute T. gondii infection. Upon oral infection, the parasite infects intestinal enterocytes, and within the lamina propria, innate immune responses lead to initial parasite control although the infection disseminates widely and persists long-term in immune privileged sites despite adaptive immunity. Upon parasite entry into the small intestine, during the acute stage, ILC1 produce high levels of IFN-γ and TNF protecting barrier surfaces, thus essentially contributing to early parasite control. We will discuss here the role of innate lymphocytes during T. gondii infection in the context of the only recently appreciated diversity of ILC subsets. © 2018 John Wiley & Sons Ltd.

  18. Antiretroviral activity of protease inhibitors against Toxoplasma gondii

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    Lianet Monzote

    2013-02-01

    Full Text Available The introduction of highly active antiretroviral therapy (HAART has caused a marked reduction in the occurrence and severity of parasitic infections, including the toxoplasmic encephalitis (TE. These changes have been attributed to the restoration of cell-mediated immunity. This study was developed to examine the activity of six antiretroviral protease inhibitors (API on Toxoplasma gondii tachyzoites. The six API showed anti-Toxoplasma activity, with IC50 value between 1.4 and 6.6 µg/mL. Further studies at the molecular level should be performed to clarify if the use of API could be beneficial or not for AIDS patients with TE.

  19. Toxoplasma gondii Infection in Seagull Chicks Is Related to the Consumption of Freshwater Food Resources

    Science.gov (United States)

    Cabezón, Oscar; Cerdà-Cuéllar, Marta; Morera, Virginia; García-Bocanegra, Ignacio; González-Solís, Jacob; Napp, Sebastian; Ribas, Maria P.; Blanch-Lázaro, Berta; Fernández-Aguilar, Xavier; Antilles, Noelia; López-Soria, Sergio; Lorca-Oró, Cristina; Dubey, Jitender P.; Almería, Sonia

    2016-01-01

    Understanding the spread of Toxoplasma gondii (T. gondii) in wild birds, particularly in those with opportunistic feeding behavior, is of interest for elucidating the epidemiological involvement of these birds in the maintenance and dissemination of the parasite. Overall, from 2009 to 2011, we collected sera from 525 seagull chicks (Yellow-legged gull (Larus michahellis) and Audouin’s gull (L. audouinii)) from 6 breeding colonies in Spain and tested them using the modified agglutination test (MAT) for the presence of antibodies against T. gondii. Chick age was estimated from bill length. Main food source of seagull chicks was evaluated using stable isotope analyses from growing scapular feathers. Overall T. gondii seroprevalence was 21.0% (IC95% 17.5–24.4). A generalized linear mixed-effects model indicated that year (2009) and food source (freshwater) were risk factors associated to the individual risk of infection by T. gondii, while age (days) was close to significance. Freshwater food origin was related to the highest seroprevalence levels, followed by marine origin, supporting freshwater and sewages as important routes of dispersion of T. gondii. Year differences could indicate fluctuating rates of exposure of seagull chicks to T. gondii. Age ranged from 4 to 30 days and seropositivity tended to increase with age (P = 0.07), supporting that seropositivity is related to T. gondii infection rather than to maternal transfer of antibodies, which in gulls is known to sharply decrease with chick age. This study is the first to report T. gondii antibodies in Yellow-legged and Audouin’s gulls, thereby extending the range of intermediate hosts for this parasite and underscoring the complexity of its epidemiology. PMID:26974667

  20. Epidemiological aspects of Toxoplasma gondii infection in riverside communities in the Southern Brazilian Amazon

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    Sérgio Neto Vitaliano

    2015-06-01

    Full Text Available INTRODUCTION: Toxoplasma gondii infection is widely prevalent in humans and other animals worldwide. Information on the prevalence of T. gondii infection is scarce in some regions of Brazil, including riverside communities along the Amazon River basin. M METHODS: The prevalence of T. gondii in 231 people, aged 1-85 years, who were living in four riverside communities along the Purus River, Lábrea, State of Amazonas, Brazil, was determined. Antibodies against T. gondii were assayed using a commercial enzyme-linked immunosorbent assay (ELISA kit. The hearts and brains of 50 chickens, which were raised free-range in the communities, were pooled according to the community of origin and bioassayed in mice. The isolates were genotyped using polymorphisms at 12 nuclear markers (SAG1, 5' and 3'-SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico and CS3. RESULTS: The overall seroprevalence of T. gondii was 56.7% (131/231. IgG antibodies were presented by 117 (89.3% and IgM by 14 (10.7% of the 131 positive individuals. No association between age group and gender with prevalence was observed (chi-square test, p > 0.05; however, the comparison between localities showed that the seroprevalence of T. gondii was significantly lower among the individuals living in the Boca do Ituxi (p < 0.05 community. Five isolates of T. gondii were obtained in the mouse bioassay, and genotyping revealed two complete genotypes that had not been described previously and three mixed isolates. CONCLUSIONS: These results support previous findings that T. gondii population genetics are highly diverse in Brazil and that T. gondii infection is active in these riverside communities.

  1. Chronic Toxoplasma gondii in Nurr1-null heterozygous mice exacerbates elevated open field activity.

    Science.gov (United States)

    Eells, Jeffrey B; Varela-Stokes, Andrea; Guo-Ross, Shirley X; Kummari, Evangel; Smith, Holly M; Cox, Erin; Lindsay, David S

    2015-01-01

    Latent infection with Toxoplasma gondii is common in humans (approximately 30% of the global population) and is a significant risk factor for schizophrenia. Since prevalence of T. gondii infection is far greater than prevalence of schizophrenia (0.5-1%), genetic risk factors are likely also necessary to contribute to schizophrenia. To test this concept in an animal model, Nurr1-null heterozygous (+/-) mice and wild-type (+/+) mice were evaluate using an emergence test, activity in an open field and with a novel object, response to bobcat urine and prepulse inhibition of the acoustic startle response (PPI) prior to and 6 weeks after infection with T. gondii. In the emergence test, T. gondii infection significantly decreased the amount of time spent in the cylinder. Toxoplasma gondii infection significantly elevated open field activity in both +/+ and +/- mice but this increase was significantly exacerbated in +/- mice. T. gondii infection reduced PPI in male +/- mice but this was not statistically significant. Aversion to bobcat urine was abolished by T. gondii infection in +/+ mice. In female +/- mice, aversion to bobcat urine remained after T. gondii infection while the male +/- mice showed no aversion to bobcat urine. Antibody titers of infected mice were a critical variable associated with changes in open field activity, such that an inverted U shaped relationship existed between antibody titers and the percent change in open field activity with a significant increase in activity at low and medium antibody titers but no effect at high antibody titers. These data demonstrate that the Nurr1 +/- genotype predisposes mice to T. gondii-induced alterations in behaviors that involve dopamine neurotransmission and are associated with symptoms of schizophrenia. We propose that these alterations in murine behavior were due to further exacerbation of the altered dopamine neurotransmission in Nurr1 +/- mice.

  2. Chronic Toxoplasma gondii in Nurr1-null heterozygous mice exacerbates elevated open field activity.

    Directory of Open Access Journals (Sweden)

    Jeffrey B Eells

    Full Text Available Latent infection with Toxoplasma gondii is common in humans (approximately 30% of the global population and is a significant risk factor for schizophrenia. Since prevalence of T. gondii infection is far greater than prevalence of schizophrenia (0.5-1%, genetic risk factors are likely also necessary to contribute to schizophrenia. To test this concept in an animal model, Nurr1-null heterozygous (+/- mice and wild-type (+/+ mice were evaluate using an emergence test, activity in an open field and with a novel object, response to bobcat urine and prepulse inhibition of the acoustic startle response (PPI prior to and 6 weeks after infection with T. gondii. In the emergence test, T. gondii infection significantly decreased the amount of time spent in the cylinder. Toxoplasma gondii infection significantly elevated open field activity in both +/+ and +/- mice but this increase was significantly exacerbated in +/- mice. T. gondii infection reduced PPI in male +/- mice but this was not statistically significant. Aversion to bobcat urine was abolished by T. gondii infection in +/+ mice. In female +/- mice, aversion to bobcat urine remained after T. gondii infection while the male +/- mice showed no aversion to bobcat urine. Antibody titers of infected mice were a critical variable associated with changes in open field activity, such that an inverted U shaped relationship existed between antibody titers and the percent change in open field activity with a significant increase in activity at low and medium antibody titers but no effect at high antibody titers. These data demonstrate that the Nurr1 +/- genotype predisposes mice to T. gondii-induced alterations in behaviors that involve dopamine neurotransmission and are associated with symptoms of schizophrenia. We propose that these alterations in murine behavior were due to further exacerbation of the altered dopamine neurotransmission in Nurr1 +/- mice.

  3. Cyclophilin A enhances cell proliferation and tumor growth of liver fluke-associated cholangiocarcinoma

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    Sawanyawisuth Kanlayanee

    2011-08-01

    Full Text Available Abstract Background Cyclophilin A (CypA expression is associated with malignant phenotypes in many cancers. However, the role and mechanisms of CypA in liver fluke-associated cholangiocarcinoma (CCA are not presently known. In this study, we investigated the expression of CypA in CCA tumor tissues and CCA cell lines as well as regulation mechanisms of CypA in tumor growth using CCA cell lines. Methods CypA expression was determined by real time RT-PCR, Western blot or immunohistochemistry. CypA silence or overexpression in CCA cells was achieved using gene delivery techniques. Cell proliferation was assessed using MTS assay or Ki-67 staining. The effect of silencing CypA on CCA tumor growth was determined in nude mice. The effect of CypA knockdown on ERK1/2 activation was assessed by Western blot. Results CypA was upregulated in 68% of CCA tumor tissues. Silencing CypA significantly suppressed cell proliferation in several CCA cell lines. Likewise, inhibition of CypA peptidyl-prolyl cis-trans isomerase (PPIase activity using cyclosporin A (CsA decreased cell proliferation. In contrast, overexpression of CypA resulted in 30% to 35% increases in proliferation of CCA cell lines. Interestingly, neither silence nor overexpression of CypA affected cell proliferation of a non-tumor human cholangiocyte cell line, MMNK1. Suppression of CypA expression attenuated ERK1/2 activity in CCA M139 cells by using both transient and stable knockdown methods. In the in vivo study, there was a 43% reduction in weight of tumors derived from CypA-silenced CCA cell lines compared with control vector CCA tumors in mice; these tumors with stable CypA silencing showed a reduced cell proliferation. Conclusions CypA is upregulated in majority of CCA patients' tissues and confers a significant growth advantage in CCA cells. Suppression of CypA expression decreases proliferation of CCA cell lines in vitro and reduces tumor growth in the nude mouse model. Inhibition of Cyp

  4. Elevated Serum Cyclophilin B Levels Are Associated with the Prevalence and Severity of Metabolic Syndrome

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    Hang Zhang

    2017-12-01

    Full Text Available ObjectiveInflammation plays a central role in the pathogenesis of metabolic syndrome (MetS. Cyclophilin B (CypB can be constitutively secreted in response to inflammatory stimuli and oxidative stress, participating in tissue or systemic inflammation. We investigated the relationship between CypB and MetS in both humans and mice.MethodsSerum CypB levels were determined in 211 subjects with MetS and 292 subjects without MetS (non-MetS (133 healthy controls and 159 high-risk subjects with one to two MetS components. Additionally, CypB expression in metabolic organs was examined in mice fed with high-fat diet (HFD and genetically obese (ob/ob mice.ResultsSerum CypB level was significantly higher in MetS subjects compared with both groups of non-MetS subjects (193.80 ± 83.22 vs. 168.38 ± 65.01 vs. 124.26 ± 47.83 ng/mL, P < 0.001. Particularly, serum CypB level was significantly higher in subjects with hypertension, central obesity, diabetes mellitus or hyperglycemia, elevated levels of triglycerides, or reduced levels of high-density lipoprotein than in those without. Moreover, CypB was positively associated with the number of MetS components (r = 0.404, P < 0.001, indicating that a higher serum CypB level reflected more severe MetS. Multivariate regression revealed that a one SD increase in CypB was associated with an odds ratio of 1.506 (1.080–2.101, P = 0.016 for MetS prevalence after adjusting for age, gender, conventional risk factors, and medication. Stratified analyses by age and gender demonstrated that subjects >60 years old with higher CypB levels were more likely to have MetS, and the risk for MetS was higher and more significant in women compared with men. Additionally, CypB expression levels were lower at baseline and dramatically enhanced in metabolic organs (such as the liver and visceral and subcutaneous adipose tissue from HFD-induced obese mice and ob/ob mice.ConclusionIncreased CypB levels were

  5. Elevated Serum Cyclophilin B Levels Are Associated with the Prevalence and Severity of Metabolic Syndrome.

    Science.gov (United States)

    Zhang, Hang; Fan, Qin; Xie, Hongyang; Lu, Lin; Tao, Rong; Wang, Fang; Xi, Rui; Hu, Jian; Chen, Qiujing; Shen, Weifeng; Zhang, Ruiyan; Yan, Xiaoxiang

    2017-01-01

    Inflammation plays a central role in the pathogenesis of metabolic syndrome (MetS). Cyclophilin B (CypB) can be constitutively secreted in response to inflammatory stimuli and oxidative stress, participating in tissue or systemic inflammation. We investigated the relationship between CypB and MetS in both humans and mice. Serum CypB levels were determined in 211 subjects with MetS and 292 subjects without MetS (non-MetS) (133 healthy controls and 159 high-risk subjects with one to two MetS components). Additionally, CypB expression in metabolic organs was examined in mice fed with high-fat diet (HFD) and genetically obese (ob/ob) mice. Serum CypB level was significantly higher in MetS subjects compared with both groups of non-MetS subjects (193.80 ± 83.22 vs. 168.38 ± 65.01 vs. 124.26 ± 47.83 ng/mL, P  CypB level was significantly higher in subjects with hypertension, central obesity, diabetes mellitus or hyperglycemia, elevated levels of triglycerides, or reduced levels of high-density lipoprotein than in those without. Moreover, CypB was positively associated with the number of MetS components ( r  = 0.404, P  CypB level reflected more severe MetS. Multivariate regression revealed that a one SD increase in CypB was associated with an odds ratio of 1.506 (1.080-2.101, P  = 0.016) for MetS prevalence after adjusting for age, gender, conventional risk factors, and medication. Stratified analyses by age and gender demonstrated that subjects >60 years old with higher CypB levels were more likely to have MetS, and the risk for MetS was higher and more significant in women compared with men. Additionally, CypB expression levels were lower at baseline and dramatically enhanced in metabolic organs (such as the liver) and visceral and subcutaneous adipose tissue from HFD-induced obese mice and ob/ob mice. Increased CypB levels were significantly and independently associated with the presence and severity of MetS, indicating that CypB could be

  6. Serosurvey of Leptospira spp. and Toxoplasma gondii in rats captured from two zoos in Southern Brazil.

    Science.gov (United States)

    Pellizzaro, Maysa; Conrado, Francisco de Oliveira; Martins, Camila Marinelli; Joaquim, Sâmea Fernandes; Ferreira, Fernando; Langoni, Helio; Biondo, Alexander Welker

    2017-01-01

    Norway rats (Rattus norvegicus) are zoonotic reservoirs for Leptospira spp. and Toxoplasma gondii, and influence diseases in urban areas. Free-ranging and laboratory-raised rats from two zoos in southern Brazil were tested for Leptospira spp. and T. gondii using microscopic agglutination and modified agglutination tests, respectively. Overall, 25.6% and 4.6% free-ranging rats tested positive for Leptospira spp. and T. gondii, respectively, with co-seropositivity occurring in two animals. For laboratory-raised rats, 20% tested positive for Leptospira spp. Also, Leptospira biflexa serovar Patoc and Leptospira noguchii serovar Panama were found. Serosurveys can show the environmental prevalence of zoonotic pathogens.

  7. Alterações cognitivas em ratos infectados com Toxoplasma gondii

    OpenAIRE

    Maia, Raquel da Silveira

    2012-01-01

    O Toxoplasma gondii é um protozoário parasito que induz alterações comportamentais em roedores. O objetivo do presente estudo foi avaliar o efeito da infecção pelo T.gondii durante a fase crônica na memória operacional e na impulsividade de roedores, bem como o efeito de antipsicóticos em reverter as eventuais alterações comportamentais decorrentes da infecção. Ratos wistar fêmeas (n=40) foram infectadas com 25 cistos da cepa ME-49 do T. gondii, após 4 meses os animais foram submetidos aos te...

  8. Seroepidemiology of infection with Toxoplasma gondii in healthy blood donors of Durango, Mexico

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    Estrada-Martínez Sergio

    2007-07-01

    Full Text Available Abstract Background Toxoplasma gondii (T. gondii infection in blood donors could represent a risk for transmission in blood recipients. There is scarce information about the epidemiology of T. gondii infection in blood donors in Mexico. Therefore, we sought to determine the prevalence of T. gondii infection and associated socio-demographic and behavioral characteristics in a population of healthy blood donors of Durango City, Mexico. Methods Four hundred and thirty two blood donors in two public blood banks of Durango City, Mexico were examined for T. gondii infection between August to September 2006. Blood donors were tested for anti-T. gondii IgG and IgM antibodies by using enzyme-linked immunoassays (Diagnostic Automation Inc., Calabasas, CA, USA. Socio-demographic and behavioral characteristics from each participant were also obtained. Results Thirty two (7.4% of 432 blood donors had IgG anti-T. gondii antibodies. Eight (1.9% of them had also IgM anti-T. gondii antibodies. Multivariate analysis using logic regression showed that T. gondii infection was associated with the presence of cats at home (adjusted OR = 3.81; 95% CI: 1.45–10.01. The age group of 45–60 years showed a significantly higher frequency of T. gondii infection than the group of 25–34 years (p = 0.02. Blood donors without education had a significantly higher frequency of infection (15.8% than those with 13–19 years of education (4.5% (p = 0.04. Other characteristics of blood donors including male gender, consumption of undercooked meat or blood transfusion did not show an association with infection. Conclusion The prevalence of T. gondii infection in healthy blood donors of Durango City, Mexico is lower than those reported in blood donors of south and central Mexico, and is one of the lowest reported in blood donors worldwide. T. gondii infection in our blood donors was most likely acquired by contact with cats. Prevalence of infection increased with age and decreased

  9. Seroprevalence of Toxoplasma gondii in Danish farmed mink (Mustela vison S.)

    DEFF Research Database (Denmark)

    Henriksen, P; Dietz, H. H.; Uttenthal, Åse

    1994-01-01

    One hundred and ninety-five mink sera randomly selected from 17 Danish mink farms were evaluated for the presence of Toxoplasma gondii antibodies in the latex agglutination test. Six (3%) sera contained T. gondii antibodies in titres of 1:64 or more. The estimated 3% prevalence means that 300 000...... mink out of a total mink population of ten million might be infected with Toxoplasma gondii. This large number of possible sero-positive mink in Denmark indicates that there exists a potential risk of acquiring toxoplasmosis by pelting mink....

  10. Antibody reaction of human anti-Toxoplasma gondii positive and negative sera with Neospora caninum antigens

    OpenAIRE

    Nam, Ho-Woo; Kang, Seung-Won; Choi, Won-Young

    1998-01-01

    Anti-Neospora caninum antibody was detected in anti-Toxoplasma gondii positive and negative human sera by ELISA, western blot and immunofluorescence assay (IFA). Twelve cases out of 172 (6.7%) Toxoplasma-positive sera cross-reacted with both T. gondii and N. caninum antigens, and one out of 110 Toxoplasma-negative sera reacted with N. caninum antigen by ELISA. By western blot, all 12 sera reacted with T. gondii antigens with various banding patterns but specifically at 30 kDa (SAG1) and 22 kD...

  11. Essential role of cyclophilin A for hepatitis C virus replication and virus production and possible link to polyprotein cleavage kinetics.

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    Artur Kaul

    2009-08-01

    Full Text Available Viruses are obligate intracellular parasites and therefore their replication completely depends on host cell factors. In case of the hepatitis C virus (HCV, a positive-strand RNA virus that in the majority of infections establishes persistence, cyclophilins are considered to play an important role in RNA replication. Subsequent to the observation that cyclosporines, known to sequester cyclophilins by direct binding, profoundly block HCV replication in cultured human hepatoma cells, conflicting results were obtained as to the particular cyclophilin (Cyp required for viral RNA replication and the underlying possible mode of action. By using a set of cell lines with stable knock-down of CypA or CypB, we demonstrate in the present work that replication of subgenomic HCV replicons of different genotypes is reduced by CypA depletion up to 1,000-fold whereas knock-down of CypB had no effect. Inhibition of replication was rescued by over-expression of wild type CypA, but not by a mutant lacking isomerase activity. Replication of JFH1-derived full length genomes was even more sensitive to CypA depletion as compared to subgenomic replicons and virus production was completely blocked. These results argue that CypA may target an additional viral factor outside of the minimal replicase contributing to RNA amplification and assembly, presumably nonstructural protein 2. By selecting for resistance against the cyclosporine analogue DEBIO-025 that targets CypA in a dose-dependent manner, we identified two mutations (V2440A and V2440L close to the cleavage site between nonstructural protein 5A and the RNA-dependent RNA polymerase in nonstructural protein 5B that slow down cleavage kinetics at this site and reduce CypA dependence of viral replication. Further amino acid substitutions at the same cleavage site accelerating processing increase CypA dependence. Our results thus identify an unexpected correlation between HCV polyprotein processing and CypA dependence

  12. Essential role of cyclophilin A for hepatitis C virus replication and virus production and possible link to polyprotein cleavage kinetics.

    Science.gov (United States)

    Kaul, Artur; Stauffer, Sarah; Berger, Carola; Pertel, Thomas; Schmitt, Jennifer; Kallis, Stephanie; Zayas, Margarita; Lopez, Margarita Zayas; Lohmann, Volker; Luban, Jeremy; Bartenschlager, Ralf

    2009-08-01

    Viruses are obligate intracellular parasites and therefore their replication completely depends on host cell factors. In case of the hepatitis C virus (HCV), a positive-strand RNA virus that in the majority of infections establishes persistence, cyclophilins are considered to play an important role in RNA replication. Subsequent to the observation that cyclosporines, known to sequester cyclophilins by direct binding, profoundly block HCV replication in cultured human hepatoma cells, conflicting results were obtained as to the particular cyclophilin (Cyp) required for viral RNA replication and the underlying possible mode of action. By using a set of cell lines with stable knock-down of CypA or CypB, we demonstrate in the present work that replication of subgenomic HCV replicons of different genotypes is reduced by CypA depletion up to 1,000-fold whereas knock-down of CypB had no effect. Inhibition of replication was rescued by over-expression of wild type CypA, but not by a mutant lacking isomerase activity. Replication of JFH1-derived full length genomes was even more sensitive to CypA depletion as compared to subgenomic replicons and virus production was completely blocked. These results argue that CypA may target an additional viral factor outside of the minimal replicase contributing to RNA amplification and assembly, presumably nonstructural protein 2. By selecting for resistance against the cyclosporine analogue DEBIO-025 that targets CypA in a dose-dependent manner, we identified two mutations (V2440A and V2440L) close to the cleavage site between nonstructural protein 5A and the RNA-dependent RNA polymerase in nonstructural protein 5B that slow down cleavage kinetics at this site and reduce CypA dependence of viral replication. Further amino acid substitutions at the same cleavage site accelerating processing increase CypA dependence. Our results thus identify an unexpected correlation between HCV polyprotein processing and CypA dependence of HCV

  13. Toxoplasma gondii: II. Tachyzoite antigenic characterization of eigth strains

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    Regina Mitsuka

    1998-01-01

    Full Text Available Eight Toxoplasma gondii strains were analyzed using ELISA and Western blot techniques, in order to demonstrate possible immunological differences. The analyzed strains were: LIV IV, LIV V and S 11 isolated from swine, RH and VPS from a human being, AS 28 from a wild mouse, HV III from a dog and CN from a cat. With the ELISA assay the eight strains showed similar reactivity with homologous and heterologous sera. The antigenic suspension, consisting of total cellular extract of tachyzoites, was effective in the indirect ELISA assay, with the positive sera reacting strongly and the negative not reacting with the antigens. The Western blot analysis showed that the T. gondii strains have similar antigenic profiles with a few variations. Three bands were observed in all strains: one of about 33 kDa (p33, another of 54 kDa (p54 and a third one of 66 kDa (p66. The HV III strain, isolated from a dog, did not show three antigens (50, 70 and 75 kDa that were present in the others. However, this difference was not detected by the ELISA assay. Only two antigens (62 kDa of the CN and 67 kDa of the LIV IV were strain-specific antigens.

  14. Risk factors for Toxoplasma gondii infection in Kohat District, Pakistan

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    Gul Naila

    2017-01-01

    Full Text Available Toxoplasma gondii is a widespread zoonotic parasite that is the causative agent for toxoplasmosis in human and completes its life cycle in separate hosts. Considering the significance of the infection, the current study was designed to asses to various risk factors for the parasite transmission to human in Kohat District, Pakistan. A total of 122 suspected individuals were asked to fill pre-designed questionnaire. A total of 44 (36.07% individuals were found to be infected with Toxoplasma gondii. Handling pets and birds, untreated water, unpasteurized milk and undercooked meat were found to be associated with infection. Raw vegetables and blood transfusion were not found to be associated with infection in our study. Thus, it can be concluded that Toxoplasma gondiiis is a prevalent zoonotic agent in Kohat, Pakistan and various prophylactic measures like hand washing, cooking meat properly, wearing gloves while contacting pets and birds, treating water and pasteurizing milk will be very helpful to reduce disease burden in the study area.

  15. Calcium uptake and proton transport by acidocalcisomes of Toxoplasma gondii.

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    Peter Rohloff

    Full Text Available Acidocalcisomes are acidic calcium stores found in diverse organisms, being conserved from bacteria to humans. They possess an acidic matrix that contains several cations bound to phosphates, which are mainly present in the form of short and long polyphosphate chains. Their matrix is acidified through the action of proton pumps such as a vacuolar proton ATPase and a vacuolar proton pyrophosphatase. Calcium uptake occurs through a Ca(2+/H(+ countertransporting ATPase located in the membrane of the organelle. Acidocalcisomes have been identified in a variety of microorganisms, including Apicomplexan parasites such as Plasmodium and Eimeria species, and in Toxoplasma gondii. We report the purification and characterization of an acidocalcisome fraction from T. gondii tachyzoites after subcellular fractionation and further discontinuous iodixanol gradient purification. Proton and calcium transport activities in the fraction were characterized by fluorescence microscopy and spectrophotometric methods using acridine orange and arsenazo III, respectively. This work will facilitate the understanding of the function of acidocalcisomes in Apicomplexan parasites, as we can now isolate highly purified fractions that could be used for proteomic analysis to find proteins that may clarify the biogenesis of these organelles.

  16. Extracellular and Intracellular Cyclophilin A, Native and Post-Translationally Modified, Show Diverse and Specific Pathological Roles in Diseases.

    Science.gov (United States)

    Xue, Chao; Sowden, Mark P; Berk, Bradford C

    2018-05-01

    CypA (cyclophilin A) is a ubiquitous and highly conserved protein with peptidyl prolyl isomerase activity. Because of its highly abundant level in the cytoplasm, most studies have focused on the roles of CypA as an intracellular protein. However, emerging evidence suggests an important role for extracellular CypA in the pathogenesis of several diseases through receptor (CD147 or other)-mediated autocrine and paracrine signaling pathways. In this review, we will discuss the shared and unique pathological roles of extracellular and intracellular CypA in human cardiovascular diseases. In addition, the evolving role of post-translational modifications of CypA in the pathogenesis of disease is discussed. Finally, recent studies with drugs specific for extracellular CypA show its importance in disease pathogenesis in several animal models and make extracellular CypA a new therapeutic target. © 2018 American Heart Association, Inc.

  17. Chemotactic Activity of Cyclophilin A in the Skin Mucus of Yellow Catfish (Pelteobagrus fulvidraco) and Its Active Site for Chemotaxis

    Science.gov (United States)

    Dawar, Farman Ullah; Tu, Jiagang; Xiong, Yang; Lan, Jiangfeng; Dong, Xing Xing; Liu, Xiaoling; Khattak, Muhammad Nasir Khan; Mei, Jie; Lin, Li

    2016-01-01

    Fish skin mucus is a dynamic barrier for invading pathogens with a variety of anti-microbial enzymes, including cyclophilin A (CypA), a multi-functional protein with peptidyl-prolyl cis/trans isomerase (PPIase) activity. Beside various other immunological functions, CypA induces leucocytes migration in vitro in teleost. In the current study, we have discovered several novel immune-relevant proteins in yellow catfish skin mucus by mass spectrometry (MS). The CypA present among them was further detected by Western blot. Moreover, the CypA present in the skin mucus displayed strong chemotactic activity for yellow catfish leucocytes. Interestingly, asparagine (like arginine in mammals) at position 69 was the critical site in yellow catfish CypA involved in leucocyte attraction. These novel efforts do not only highlight the enzymatic texture of skin mucus, but signify CypA to be targeted for anti-inflammatory therapeutics. PMID:27589721

  18. Chemotactic Activity of Cyclophilin A in the Skin Mucus of Yellow Catfish (Pelteobagrus fulvidraco and Its Active Site for Chemotaxis

    Directory of Open Access Journals (Sweden)

    Farman Ullah Dawar

    2016-08-01

    Full Text Available Fish skin mucus is a dynamic barrier for invading pathogens with a variety of anti-microbial enzymes, including cyclophilin A (CypA, a multi-functional protein with peptidyl-prolyl cis/trans isomerase (PPIase activity. Beside various other immunological functions, CypA induces leucocytes migration in vitro in teleost. In the current study, we have discovered several novel immune-relevant proteins in yellow catfish skin mucus by mass spectrometry (MS. The CypA present among them was further detected by Western blot. Moreover, the CypA present in the skin mucus displayed strong chemotactic activity for yellow catfish leucocytes. Interestingly, asparagine (like arginine in mammals at position 69 was the critical site in yellow catfish CypA involved in leucocyte attraction. These novel efforts do not only highlight the enzymatic texture of skin mucus, but signify CypA to be targeted for anti-inflammatory therapeutics.

  19. Overexpressed cyclophilin B suppresses aldosterone-induced proximal tubular cell injury both in vitro and in vivo.

    Science.gov (United States)

    Wang, Bin; Lin, Lilu; Wang, Haidong; Guo, Honglei; Gu, Yong; Ding, Wei

    2016-10-25

    The renin-angiotensin-aldosterone system (RAAS) is overactivated in patients with chronic kidney disease. Oxidative stress and endoplasmic reticulum stress (ERS) are two major mechanisms responsible for aldosterone-induced kidney injury. Cyclophilin (CYP) B is a chaperone protein that accelerates the rate of protein folding through its peptidyl-prolyl cis-trans isomerase (PPIase) activity. We report that overexpression of wild-type CYPB attenuated aldosterone-induced oxidative stress (evidenced by reduced production of reactive oxygen species and improved mitochondrial dysfunction), ERS (indicated by reduced expression of the ERS markers glucose-regulated protein 78 [GRP78] and C/-EBP homologous protein [CHOP]), and tubular cell apoptosis in comparison with aldosterone-induced human kidney-2 (HK-2) cells. The in vivo study also yielded similar results. Hence, CYPB performs a crucial function in protecting cells against aldosterone-induced oxidative stress, ERS, and tubular cell injury via its PPIase activity.

  20. Molecular analysis of Toxoplasma gondii Surface Antigen 1 (SAG1) gene cloned from Toxoplasma gondii DNA isolated from Javanese acute toxoplasmosis

    Science.gov (United States)

    Haryati, Sri; Agung Prasetyo, Afiono; Sari, Yulia; Dharmawan, Ruben

    2018-05-01

    Toxoplasma gondii Surface Antigen 1 (SAG1) is often used as a diagnostic tool due to its immunodominant-specific as antigen. However, data of the Toxoplasma gondii SAG1 protein from Indonesian isolate is limited. To study the protein, genomic DNA was isolated from a Javanese acute toxoplasmosis blood samples patient. A complete coding sequence of Toxoplasma gondii SAG1 was cloned and inserted into an Escherichia coli expression plasmid and sequenced. The sequencing results were subjected to bioinformatics analysis. The Toxoplasma gondii SAG1 complete coding sequences were successfully cloned. Physicochemical analysis revealed the 336 aa of SAG1 had 34.7 kDa of weight. The isoelectric point and aliphatic index were 8.4 and 78.4, respectively. The N-terminal methionine half-life in Escherichia coli was more than 10 hours. The antigenicity, secondary structure, and identification of the HLA binding motifs also had been discussed. The results of this study would contribute information about Toxoplasma gondii SAG1 and benefits for further works willing to develop diagnostic and therapeutic strategies against the parasite.

  1. Discovery of non-peptidic small molecule inhibitors of cyclophilin D as neuroprotective agents in Aβ-induced mitochondrial dysfunction

    Science.gov (United States)

    Park, Insun; Londhe, Ashwini M.; Lim, Ji Woong; Park, Beoung-Geon; Jung, Seo Yun; Lee, Jae Yeol; Lim, Sang Min; No, Kyoung Tai; Lee, Jiyoun; Pae, Ae Nim

    2017-10-01

    Cyclophilin D (CypD) is a mitochondria-specific cyclophilin that is known to play a pivotal role in the formation of the mitochondrial permeability transition pore (mPTP).The formation and opening of the mPTP disrupt mitochondrial homeostasis, cause mitochondrial dysfunction and eventually lead to cell death. Several recent studies have found that CypD promotes the formation of the mPTP upon binding to β amyloid (Aβ) peptides inside brain mitochondria, suggesting that neuronal CypD has a potential to be a promising therapeutic target for Alzheimer's disease (AD). In this study, we generated an energy-based pharmacophore model by using the crystal structure of CypD—cyclosporine A (CsA) complex and performed virtual screening of ChemDiv database, which yielded forty-five potential hit compounds with novel scaffolds. We further tested those compounds using mitochondrial functional assays in neuronal cells and identified fifteen compounds with excellent protective effects against Aβ-induced mitochondrial dysfunction. To validate whether these effects derived from binding to CypD, we performed surface plasmon resonance (SPR)—based direct binding assays with selected compounds and discovered compound 29 was found to have the equilibrium dissociation constants (KD) value of 88.2 nM. This binding affinity value and biological activity correspond well with our predicted binding mode. We believe that this study offers new insights into the rational design of small molecule CypD inhibitors, and provides a promising lead for future therapeutic development.

  2. Keratinocyte secretion of cyclophilin B via the constitutive pathway is regulated through its cyclosporin-binding site.

    Science.gov (United States)

    Fearon, Paula; Lonsdale-Eccles, Ann A; Ross, O Kehinde; Todd, Carole; Sinha, Aparna; Allain, Fabrice; Reynolds, Nick J

    2011-05-01

    Cyclophilin B (CypB) is an endoplasmic reticulum (ER)-resident member of the cyclophilin family of proteins that bind cyclosporin A (CsA). We report that as in other cell types, CypB trafficked from the ER and was secreted by keratinocytes into the media in response to CsA. Concentrations as low as 1 pM of CsA induced secretion of CypB. Using brefeldin A, we showed that CypB is secreted from keratinocytes via the constitutive secretory pathway. We defined that substitution of tryptophan residue 128 in the CsA-binding site of CypB with alanine resulted in dissociation of CypB(W128A)-green fluorescent protein (GFP) from the ER. Photobleaching studies revealed a significant reduction in the diffusible mobility of CypB(W128A)-GFP compared with CypB(WT)-GFP, consistent with redistribution of CypB(W128A)-GFP into secretory vesicles disconnected from the ER/Golgi network. Furthermore, CsA significantly decreased the mobility of CypB(WT)-GFP but not CypB(W128A)-GFP. These studies demonstrate that therapeutically relevant concentrations of CsA regulate secretion of CypB by keratinocytes, and that a key residue within the CsA-binding site of CypB controls retention of CypB within the ER and regulates entry into the secretory pathway. As keratinocytes express CypB receptors (CD147) and CypB exhibits chemotactic properties, these data have implications for the therapeutic effects of CsA in inflammatory skin disease.

  3. Involvement of two classes of binding sites in the interactions of cyclophilin B with peripheral blood T-lymphocytes.

    Science.gov (United States)

    Denys, A; Allain, F; Carpentier, M; Spik, G

    1998-12-15

    Cyclophilin B (CyPB) is a cyclosporin A (CsA)-binding protein, mainly associated with the secretory pathway, and is released in biological fluids. We recently reported that CyPB specifically binds to T-lymphocytes and promotes enhanced incorporation of CsA. The interactions with cellular binding sites involved, at least in part, the specific N-terminal extension of the protein. In this study, we intended to specify further the nature of the CyPB-binding sites on peripheral blood T-lymphocytes. We first provide evidence that the CyPB binding to heparin-Sepharose is prevented by soluble sulphated glycosaminoglycans (GAG), raising the interesting possibility that such interactions may occur on the T-cell surface. We then characterized CyPB binding to T-cell surface GAG and found that these interactions involved the N-terminal extension of CyPB, but not its conserved CsA-binding domain. In addition, we determined the presence of a second CyPB binding site, which we termed a type I site, in contrast with type II for GAG interactions. The two binding sites exhibit a similar affinity but the expression of the type I site was 3-fold lower. The conclusion that CyPB binding to the type I site is distinct from the interactions with GAG was based on the findings that it was (1) resistant to NaCl wash and GAG-degrading enzyme treatments, (2) reduced in the presence of CsA or cyclophilin C, and (3) unmodified in the presence of either the N-terminal peptide of CyPB or protamine. Finally, we showed that the type I binding sites were involved in an endocytosis process, supporting the hypothesis that they may correspond to a functional receptor for CyPB.

  4. Ultra-violet B (UVB)-induced skin cell death occurs through a cyclophilin D intrinsic signaling pathway

    Energy Technology Data Exchange (ETDEWEB)

    Ji, Chao [Department of Dermatology, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210024, Jiangsu (China); Yang, Bo [Department of Dermatology, Huashan Hospital, Fudan University, Shanghai 200040 (China); Yang, Zhi; Tu, Ying [Department of Dermatology, The First Affiliated Hospital of Kunming Medical University, Yunnan Provincial Institute of Dermatology, Kunming 650032, Yunnan (China); Yang, Yan-li [Department of Otorhinolaryngology, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210024, Jiangsu (China); He, Li, E-mail: heli2662@yahoo.com.cn [Department of Otorhinolaryngology, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210024, Jiangsu (China); Bi, Zhi-Gang, E-mail: eltonbibenqhospital@yahoo.com.cn [Department of Dermatology, BenQ Medical Center, Nanjing Medical University, Nanjing 210019, Jiangsu (China)

    2012-09-07

    Highlights: Black-Right-Pointing-Pointer UVB radiated skin keratinocytes show cyclophilin D (Cyp-D) upregulation. Black-Right-Pointing-Pointer NAC inhibits UVB induced Cyp-D expression, while H{sub 2}O{sub 2} facilitates it. Black-Right-Pointing-Pointer Cyp-D-deficient cells are significantly less susceptible to UVB induced cell death. Black-Right-Pointing-Pointer Over-expression of Cyp-D causes spontaneous keratinocytes cell death. -- Abstract: UVB-induced skin cell damage involves the opening of mitochondrial permeability transition pore (mPTP), which leads to both apoptotic and necrotic cell death. Cyclophilin D (Cyp-D) translocation to the inner membrane of mitochondrion acts as a key component to open the mPTP. Our Western-Blot results in primary cultured human skin keratinocytes and in HaCaT cell line demonstrated that UVB radiation and hydrogen peroxide (H{sub 2}O{sub 2}) induced Cyp-D expression, which was inhibited by anti-oxidant N-acetyl cysteine (NAC). We created a stable Cyp-D deficiency skin keratinocytes by expressing Cyp-D-shRNA through lentiviral infection. Cyp-D-deficient cells were significantly less susceptible than their counterparts to UVB- or H{sub 2}O{sub 2}-induced cell death. Further, cyclosporine A (Cs-A), a Cyp-D inhibitor, inhibited UVB- or H{sub 2}O{sub 2}-induced keratinocytes cell death. Reversely, over-expression of Cyp-D in primary keratinocytes caused spontaneous keratinocytes cell death. These results suggest Cyp-D's critical role in UVB/oxidative stress-induced skin cell death.

  5. Ultra-violet B (UVB)-induced skin cell death occurs through a cyclophilin D intrinsic signaling pathway

    International Nuclear Information System (INIS)

    Ji, Chao; Yang, Bo; Yang, Zhi; Tu, Ying; Yang, Yan-li; He, Li; Bi, Zhi-Gang

    2012-01-01

    Highlights: ► UVB radiated skin keratinocytes show cyclophilin D (Cyp-D) upregulation. ► NAC inhibits UVB induced Cyp-D expression, while H 2 O 2 facilitates it. ► Cyp-D-deficient cells are significantly less susceptible to UVB induced cell death. ► Over-expression of Cyp-D causes spontaneous keratinocytes cell death. -- Abstract: UVB-induced skin cell damage involves the opening of mitochondrial permeability transition pore (mPTP), which leads to both apoptotic and necrotic cell death. Cyclophilin D (Cyp-D) translocation to the inner membrane of mitochondrion acts as a key component to open the mPTP. Our Western-Blot results in primary cultured human skin keratinocytes and in HaCaT cell line demonstrated that UVB radiation and hydrogen peroxide (H 2 O 2 ) induced Cyp-D expression, which was inhibited by anti-oxidant N-acetyl cysteine (NAC). We created a stable Cyp-D deficiency skin keratinocytes by expressing Cyp-D-shRNA through lentiviral infection. Cyp-D-deficient cells were significantly less susceptible than their counterparts to UVB- or H 2 O 2 -induced cell death. Further, cyclosporine A (Cs-A), a Cyp-D inhibitor, inhibited UVB- or H 2 O 2 -induced keratinocytes cell death. Reversely, over-expression of Cyp-D in primary keratinocytes caused spontaneous keratinocytes cell death. These results suggest Cyp-D’s critical role in UVB/oxidative stress-induced skin cell death.

  6. First molecular evidence of Toxoplasma gondii in opossums (Didelphis virginiana from Yucatan, Mexico

    Directory of Open Access Journals (Sweden)

    M. Torres-Castro

    2016-03-01

    Full Text Available Toxoplasma gondii is an obligate intracellular parasite recognized as a causal agent of toxoplasmosis; zoonotic disease endemic in many countries worldwide, including Mexico. Different species of animals participate in the wild cycle infection, including opossums of the species Didelphis virginiana. Thirteen D. virginiana were captured in Yucatan, Mexico. Detection of T. gondii was achieved by Polymerase Chain Reaction, which determined an infection of 76.9% (10/13 in brains. Positive amplicons were sequenced for analysis, this produced results similar to T. gondii with identity and coverage values of 98% and 96-100%, respectively. This study presents the first molecular evidence of the circulation of T. gondii in D. virginiana from Mexico.

  7. First molecular evidence of Toxoplasma gondii in opossums (Didelphis virginiana) from Yucatan, Mexico.

    Science.gov (United States)

    Torres-Castro, M; Noh-Pech, H; Puerto-Hernández, R; Reyes-Hernández, B; Panti-May, A; Hernández-Betancourt, S; Yeh-Gorocica, A; González-Herrera, L; Zavala-Castro, J; Puerto, F I

    2016-01-01

    Toxoplasma gondii is an obligate intracellular parasite recognized as a causal agent of toxoplasmosis; zoonotic disease endemic in many countries worldwide, including Mexico. Different species of animals participate in the wild cycle infection, including opossums of the species Didelphis virginiana. Thirteen D. virginiana were captured in Yucatan, Mexico. Detection of T. gondii was achieved by Polymerase Chain Reaction, which determined an infection of 76.9% (10/13) in brains. Positive amplicons were sequenced for analysis, this produced results similar to T. gondii with identity and coverage values of 98% and 96-100%, respectively. This study presents the first molecular evidence of the circulation of T. gondii in D. virginiana from Mexico.

  8. Prevalence of Toxoplasma gondii antibodies and intestinal parasites in stray cats from Nigde, Turkey

    Directory of Open Access Journals (Sweden)

    Bengi Dündar

    2010-01-01

    Full Text Available The prevalence of antibodies to Toxoplasma gondii was investigated by the Sabin-Feldman Dye test (SFDT in 72 stray cats from Nigde, Turkey. A total of 55 (76.4% of the analysed sera had antibodies to T. gondii. The seropositivity of T. gondii was 77.1% in male and 75.7% in female cats (P>0.05. Faeces of these cats were also examined by zinc sulphate flotation method for the presence of parasite oocysts and eggs of other parasites. Two protozoan parasites were identified as Isospora spp. (12.5% and Eimeria spp. (4.1% in cats. Toxoplasma gondii oocysts were not found in any faecal samples analysed. Two parasitic helminth species were observed: Toxocara cati (15.2% and Toxascaris leonina (20.8%. These common ascarids were recorded for the first time in cats from Nigde.

  9. Brain cancer mortality rates increase with Toxoplasma gondii seroprevalence in France

    Science.gov (United States)

    Vittecoq, Marion; Elguero, Eric; Lafferty, Kevin D.; Roche, Benjamin; Brodeur, Jacques; Gauthier-Clerc, Michel; Missé, Dorothée; Thomas, Frédéric

    2012-01-01

    The incidence of adult brain cancer was previously shown to be higher in countries where the parasite Toxoplasma gondii is common, suggesting that this brain protozoan could potentially increase the risk of tumor formation. Using countries as replicates has, however, several potential confounding factors, particularly because detection rates vary with country wealth. Using an independent dataset entirely within France, we further establish the significance of the association between T. gondii and brain cancer and find additional demographic resolution. In adult age classes 55 years and older, regional mortality rates due to brain cancer correlated positively with the local seroprevalence of T. gondii. This effect was particularly strong for men. While this novel evidence of a significant statistical association between T. gondii infection and brain cancer does not demonstrate causation, these results suggest that investigations at the scale of the individual are merited.

  10. Environmental Exposures Are Important Risk Factors for Infection Toxoplasma gondii and Helicobacter pylori

    Science.gov (United States)

    Background: An estimated 70% of Americans suffer chronic infections. Helicobacter pylori and Toxoplasma gondii affect an estimated 35% and 15% of Americans, respectively. Despite their heavy burden, environmental transmission of these infections is not well understood. Object...

  11. 97 original article toxoplasma gondii infection in hiv/aids: prevalence

    African Journals Online (AJOL)

    Keywords: Toxoplasma gondii , IgG, Seroprevalence, HIV positive, CD4 cells. ... with immunosuppressive cancer and transplant ... due to active parasitaemia during pregnancy can .... especially in the night where they look for leftover.

  12. Seroprevalence of Toxoplasma gondii from free-ranging black bears ( Ursus americanus ) from Florida.

    Science.gov (United States)

    Chambers, D L; Ulrey, W A; Guthrie, J M; Kwok, O C H; Cox, J J; Maehr, D S; Dubey, J P

    2012-06-01

    Toxoplasma gondii is a significant worldwide parasitic protozoan. In the present study, prevalence of antibodies of T. gondii was examined from 29 free-ranging black bears ( Ursus americanus ) from south-central Florida where the host species was listed as state threatened during this project. Overall T. gondii prevalence was found to be 44.8%, specifically 46.2% in male and 43.8% in female U. americanus , using a modified agglutination test (1:25 titer). Seroprevalence differences between sexes were not significant (P > 0.05). Results of the present study add supportive data to the growing body of evidence suggesting that U. americanus has one of the highest T. gondii seroprevalences among all known intermediate hosts. In addition, our data emphasize the importance of understanding parasitic disease dynamics from a conservation perspective.

  13. Toxoplasma gondii in horse meat intended for human consumption in Romania.

    Science.gov (United States)

    Paştiu, Anamaria Ioana; Györke, Adriana; Kalmár, Zsuzsa; Bolfă, Pompei; Rosenthal, Benjamin Martin; Oltean, Miruna; Villena, Isabelle; Spînu, Marina; Cozma, Vasile

    2015-09-15

    The prevalence of Toxoplasma gondii, an economically important zoonotic protozoan, was investigated in horses slaughtered for export and human consumption in the North of Romania. Pairs of samples, sera and heart tissues, were collected from 82 slaughtered horses. Examination of horse sera by ELISA at a dilution of 1:10, and by modified agglutination test (MAT) at a dilution of 1:6, revealed that 32 (39%) and 31(37.8%) horses, respectively, had antibodies against T. gondii. Using polymerase chain reaction (PCR) analysis, T. gondii DNA was not found in any heart sample collected from horses. By bioassay in mice, we obtained viable isolates of T. gondii from two of ten horses determined to be strongly positive by serological assay/ELISA. The prevalence estimated in horses highlighted the potential risk for human contamination by consumption of raw or undercooked meat. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Literature Reference for Toxoplasma gondii (Applied and Environmental Microbiology. 2004. 70(7): 4035–4039)

    Science.gov (United States)

    Procedures are described for analysis of water samples and may be adapted for assessment of solid, particulate and liquid samples. The method uses real-time PCR assay for detecting Toxoplasma gondii DNA using gene-specific primers and probe.

  15. Infection rate of toxoplasma gondii and age distribution in female patients with sterility

    International Nuclear Information System (INIS)

    Li Shuhong; Dai Pei; Cui Liming; Zong Shan; Zuo Wenjing

    2006-01-01

    Objective: To discuss the relationship between the infection of Toxoplasma gondii and female sterility. Methods: Toxoplasma gondii serum antibody were determined in 882 women with sterility (experimental group) and 107 normal bearing women (control group) by using ELISA. At the same time the differences of the infection with Toxoplasma gondii between the ages of the sterility women were analyzed. Results: The positive rate in experimental group was 15.87% (140/882), the positive rate in control group was 5.61% (6/107), remarkable difference was found between two groups (P<0.01). The infection rate in the different age groups (20-24, 25-29, 30-34, 35-39 and ≥40) is 5.63%, 15.24%, 17.91%, 19.44% and 15.38%. Conclusion: Toxoplasma gondii infection may be one of the factors which can cause sterility, and the infection rates at different ages have no instinct differences. (authors)

  16. Detection of toxoplasma gondii antigens in sera from experimentally infected mice

    International Nuclear Information System (INIS)

    Shojaee, S.; Keshavarz, H.; Rezaian, M.; Mohebali, M.

    2007-01-01

    Detection of Toxoplasma antigen in serum of mice by Immunoblotting. strain. IgG isolated from rabbits that were immunized with T. gondii Immunoblotting was performed to detect T. gondii antigens in sera of mice. Serum samples from mice experimentally infected with T. gondii RH strain. The value of Immunoblotting in diagnosis of toxoplasmosis in acute stage of infection. The antigen bands detected in serum sample of mice were experimentally infected with T. gondii tachyzoite in immunoblotting. Six bands demonstrated on seventh post infection day six bands were identified. Similarly on sixth day four bands, on day five three bands and on fourth post infection day two bands were identified. No band was detected in control group sera. Immunoblotting is a sensitive method for diagnosis of acute stage of toxoplasmosis. (author)

  17. Detection of Toxoplasma gondii DNA in sera samples of mice experimentally infected

    Directory of Open Access Journals (Sweden)

    H. Langoni

    2006-04-01

    Full Text Available Detection of Toxoplasma gondii (T. gondii DNA in blood can help to diagnose the disease in its acute phase; however, it must be considered that hemoglobin, present in blood, can inhibit polymerase activity, making impracticable the detection of DNA in samples. Mice were experimentally infected via oral route with ME49 and BTU2 strains cysts and RH strain tachyzoites; polymerase chain reaction was used to detect T. gondii DNA in mice sera 18, 24, 48, 96, and 192 hours post infection (PI. Toxoplama gondii DNA was detected in only one animal infected with BTU2 strain, genotype III (isolated from a dog with neurological signs 18 hours PI. The agent's DNA was not detected in any sample of the other experimental groups. New studies must be carried out to verify the technique sensitivity in researches on this agent's genetic material using sera samples of acute-phase toxoplasmosis patients, especially in cases of immunosuppression.

  18. Anti-t B toxopla Balsam asma g mocitru gondii us cam activit merune ...

    African Journals Online (AJOL)

    SAM

    s in humans igate intracel .... infertility, and influenza (Tsassi et al., 2010; Asase et al.,. 2005). .... Human foreskin fibroblasts (HFF) cells were infected by tachyzoites of T. gondii .... Prophylaxis against opportunistic infections in patients with ...

  19. The current status of Toxoplasma gondii infection among Egyptian rheumatoid arthritis patients

    Directory of Open Access Journals (Sweden)

    Nagwa Mostafa El-Sayed

    2016-10-01

    Full Text Available Objective: To ascertain a relationship between Toxoplasma gondii (T. gondii infection and rheumatoid arthritis (RA disease among Egyptian patients. Methods: One hundred RA patients and 50 healthy subjects participated in this study. The patients were classified into three groups, namely GI, G2 and G3. Patients in G1 were recently diagnosed with RA with the disease duration of less than one year (prior treatment; G2 included RA patients receiving anti-tumor necrosis factor agents and RA patients in G3 received disease modifying anti-rheumatic drugs (methotrexate, antimalarial, corticosteroids. Serum samples of all participants were examined for the presence of anti-Toxoplasma immunoglobulin G (IgG antibodies and positive samples were further analyzed for antiToxoplasma IgM antibodies to detect the possibility of reactivation of latent toxoplasmosis. Also, the association between Toxoplasma seropositivity and clinical, laboratory and radiological features of these patients were determined. Results: There was a significantly higher percentage of T. gondii IgG positivity in RA patients (54% than in the controls (32%. At the same time, 20.40% of T. gondii IgG positive patients had anti-T. gondii IgM antibodies with a statistically significant difference as comparing to T. gondii IgG positive controls. Out of T. gondii seropositive patients, 20.37% had a lower IgG level with a mean titer of (65.3 ± 17.7 IU/mL, 46.29% had moderate level with a mean titer of (184.2 ± 60.0 IU/mL and 33.33% had higher level with a mean titer of (404.3 ± 50.0 IU/ mL. A positive correlation was found between disease activity and Toxoplasma seropositivity. T. gondii seropositive RA patients had longer disease duration, longer time morning stiffness, higher numbers of tender and swollen joints and also increase in disease severity markers (erythrocyte sedimentation rate, C-reactive protein, disease activity score 28, anti-cyclic citrullinated peptide anti

  20. First report of Toxoplasma gondii seroprevalence in pet parrots in China.

    Science.gov (United States)

    Zhang, Xiao-Xuan; Zhang, Nian-Zhang; Tian, Wei-Peng; Zhou, Dong-Hui; Xu, Ying-Tian; Zhu, Xing-Quan

    2014-06-01

    Toxoplasmosis, caused by the obligate intracellular protozoan parasite Toxoplasma gondii, has become a serious public health problem worldwide. T. gondii can infect almost all warm-blooded animals, including parrots. However, little is known of T. gondii infection in parrots in China. Antibodies against T. gondii in 311 parrots including 202 Budgerigars (Melopsittacus undulatus), 26 Lovebirds (Agapornis sp.), 22 Cockatiels (Nymphicus hollandicus), and 61 Alexandrine Parakeets (Psittacula eupatria) in the cities of Beijing and Weifang in north China were tested using the modified agglutination test (MAT). Twenty-six (8.36%) out of 311 serum samples were positive for T. gondii at the cutoff of 1:5. Among the four species, a higher seroprevalence of T. gondii was found in Cockatiels (13.64%, 95% confidence interval [CI] 0.00-27.98), although the difference was not statistically significant (p=0.61). Seropositivity rates against T. gondii in male parrots (10.43%, 95% CI 5.74-15.12) were not statistically different from that in female parrots (6.08%, 95% CI 2.23-9.93, p=0.17). The seropositivity of T. gondii in parrots from Weifang and Beijing was 11.11% (95% CI 6.13-16.09) and 5.70% (95% CI 2.08-9.31), respectively. The seroprevalence varied in parrots of different age groups, ranging from 5.71% (95% CI 1.27-10.15) to 13.00% (95% CI 6.41-19.69), however, the difference among age groups was not statistically significant (p=0.12). The seroprevalence of T. gondii infection in parrots in summer (11.63%, 95% CI 6.84-16.42) was significantly higher than in spring (4.32%, 95% CI 0.94-7.70, p=0.02). The results of the present survey indicated that parrots in China are exposed to T. gondii. To our knowledge, this is the first report of T. gondii seroprevalence in parrots in China.

  1. Toxoplasma gondii infection in interstate truck drivers: a case-control seroprevalence study.

    Science.gov (United States)

    Alvarado-Esquivel, Cosme; Pacheco-Vega, Sandy Janet; Hernández-Tinoco, Jesús; Salcedo-Jáquez, Misael; Sánchez-Anguiano, Luis Francisco; Berumen-Segovia, Luis Omar; Rábago-Sánchez, Elizabeth; Liesenfeld, Oliver

    2015-02-05

    Infection with Toxoplasma gondii can be acquired via the ingestion of undercooked or raw meat containing tissue cysts, or via ingestion of water contaminated with oocysts. Professional long distance truck driving may have epidemiological importance for food-borne infections since drivers eat out of home and in places where hygiene and cooking practices are uncertain. We aimed to determine whether interstate truck drivers in Durango, Mexico have an increased risk of infection with T. gondii as indicated by seropositivity; and to determine the socio-demographic, work, clinical, and behavioral characteristics associated with T. gondii seropositivity in interstate truck drivers. Through a case-control study design, 192 truck drivers and 192 controls from the general population of the same region matched by gender and age were examined with enzyme-linked immunoassays for the presence of anti-Toxoplasma IgG and IgM antibodies. Socio-demographic, work, clinical and behavioral characteristics from the truck drivers were obtained. Anti-T. gondii IgG antibodies were found in 23 (12.0%) of 192 truck drivers and in 13 (6.8%) of 192 controls (OR = 21.0; 95% CI: 1.23-358.38; P = 0.002). Anti-T. gondii IgM antibodies were found in 7 (3.6%) cases and in 7 (3.6%) controls (P = 1.00). The seroprevalence of T. gondii infection was higher in drivers with reflex impairment than in those without this impairment (4/13, 30.8% vs 19/179, 10.6%, respectively; P = 0.05), and in drivers with hearing impairment than in those without this impairment (3/7, 42.9% vs 20/185, 10.8%, respectively; P = 0.03). Multivariate analysis of work and behavioral characteristics of truck drives showed positive associations of T. gondii exposure with trips to the south of Mexico (OR = 3.11; 95% CI: 1.02-9.44; P = 0.04) and consumption of horse meat (OR = 5.18; 95% CI: 1.62-16.55; P = 0.005). Results suggest that interstate truck drivers may have an increased risk for T

  2. First molecular evidence of Toxoplasma gondii in opossums (Didelphis virginiana) from Yucatan, Mexico

    OpenAIRE

    Torres-Castro, M.; Noh-Pech, H.; Puerto-Hern?ndez, R.; Reyes-Hern?ndez, B.; Panti-May, A.; Hern?ndez-Betancourt, S.; Yeh-Gorocica, A.; Gonz?lez-Herrera, L.; Zavala-Castro, J.; Puerto, F.I.

    2016-01-01

    Toxoplasma gondii is an obligate intracellular parasite recognized as a causal agent of toxoplasmosis; zoonotic disease endemic in many countries worldwide, including Mexico. Different species of animals participate in the wild cycle infection, including opossums of the species Didelphis virginiana. Thirteen D. virginiana were captured in Yucatan, Mexico. Detection of T. gondii was achieved by Polymerase Chain Reaction, which determined an infection of 76.9% (10/13) in brains. Positive amplic...

  3. Prevalence of Toxoplasma gondii infection in wild birds in Durango, Mexico.

    Science.gov (United States)

    Alvarado-Esquivel, C; Rajendran, C; Ferreira, L R; Kwok, O C H; Choudhary, S; Alvarado-Esquivel, D; Rodríguez-Peña, S; Villena, I; Dubey, J P

    2011-10-01

    There is a lack of information concerning the prevalence of Toxoplasma gondii infection in wild birds in Mexico. In the present study, serum samples and tissues from 653 birds from Durango State, Mexico, were evaluated for T. gondii infection. Antibodies to T. gondii (modified agglutination test, titer 1∶25 or higher) were found in 17 (2.6%) of the 653 birds, including 1 of 2 curve-billed thrashers (Toxostoma curvirostre), 2 (1 Anas platyrhynchos, 1 Anas diazi) of 4 ducks, 1 of 2 eagles (Aquila sp.), 5 (27.8%) of 18 great-tailed grackles (Quiscalus mexicanus), 7 (1.3%) of 521 rock pigeons (Columba livia), and 1 (14.3%) of 7 quail (Coturnix coturnix). The seroprevalence of T. gondii infection in birds captured in a park outside the city zoo (11.6%, 8/69) was significantly higher than that found in birds from other regions (1.5%, 9/584, OR  =  8.38; 95% CI: 2.82-24.77; P  =  0.0001). Brains and hearts of 23 birds (17 seropositive, 6 seronegative) were bioassayed in mice for the isolation of T. gondii . Viable T. gondii was isolated from 1 of 7 seropositive pigeons. The DNA obtained from the T. gondii isolate from the pigeon was genotyped using the PCR-RFLP typing using 11 markers (B1, SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico) and revealed an atypical genotype. This is the first report of T. gondii infection in great-tailed grackles, the Mexican duck, and curved-billed thrashers and the first survey of wild birds in Mexico.

  4. Epidemiology and pathology of Toxoplasma gondii in free-ranging California sea lions (Zalophus californianus).

    Science.gov (United States)

    Carlson-Bremer, Daphne; Colegrove, Kathleen M; Gulland, Frances M D; Conrad, Patricia A; Mazet, Jonna A K; Johnson, Christine K

    2015-04-01

    The coccidian parasite Toxoplasma gondii infects humans and warm-blooded animals worldwide. The ecology of this parasite in marine systems is poorly understood, although many marine mammals are infected and susceptible to clinical toxoplasmosis. We summarized the lesions associated with T. gondii infection in the California sea lion (Zalophus californianus) population and investigated the prevalence of and risk factors associated with T. gondii exposure, as indicated by antibody. Five confirmed and four suspected cases of T. gondii infection were identified by analysis of 1,152 medical records of necropsied sea lions from 1975-2009. One suspected and two confirmed cases were identified in aborted fetuses from a sea lion rookery. Toxoplasmosis was the primary cause of death in five cases, including the two fetuses. Gross and histopathologic findings in T. gondii-infected sea lions were similar to those reported in other marine mammals. The most common lesions were encephalitis, meningitis, and myocarditis. The antibody prevalence in stranded, free-ranging sea lions for 1998-2009 was 2.5% (±0.03%; IgG titer 640). There was an increase in odds of exposure in sea lions with increasing age, suggesting cumulative risk of exposure and persistent antibody over time. The occurrence of disseminated T. gondii infection in aborted fetuses confirms vertical transmission in sea lions, and the increasing odds of exposure with age is consistent with additional opportunities for horizontal transmission in free-ranging sea lions over time. These data suggest that T. gondii may have two modes of transmission in the sea lion population. Overall, clinical disease was uncommon in our study which, along with low prevalence of T. gondii antibody, suggests substantially less-frequent exposure and lower susceptibility to clinical disease in California sea lions as compared to sympatric southern sea otters (Enhydra lutris nereis).

  5. The common parasite Toxoplasma gondii induces prostatic inflammation and microglandular hyperplasia in a mouse model.

    Science.gov (United States)

    Colinot, Darrelle L; Garbuz, Tamila; Bosland, Maarten C; Wang, Liang; Rice, Susan E; Sullivan, William J; Arrizabalaga, Gustavo; Jerde, Travis J

    2017-07-01

    Inflammation is the most prevalent and widespread histological finding in the human prostate, and associates with the development and progression of benign prostatic hyperplasia and prostate cancer. Several factors have been hypothesized to cause inflammation, yet the role each may play in the etiology of prostatic inflammation remains unclear. This study examined the possibility that the common protozoan parasite Toxoplasma gondii induces prostatic inflammation and reactive hyperplasia in a mouse model. Male mice were infected systemically with T. gondii parasites and prostatic inflammation was scored based on severity and focality of infiltrating leukocytes and epithelial hyperplasia. We characterized inflammatory cells with flow cytometry and the resulting epithelial proliferation with bromodeoxyuridine (BrdU) incorporation. We found that T. gondii infects the mouse prostate within the first 14 days of infection and can establish parasite cysts that persist for at least 60 days. T. gondii infection induces a substantial and chronic inflammatory reaction in the mouse prostate characterized by monocytic and lymphocytic inflammatory infiltrate. T. gondii-induced inflammation results in reactive hyperplasia, involving basal and luminal epithelial proliferation, and the exhibition of proliferative inflammatory microglandular hyperplasia in inflamed mouse prostates. This study identifies the common parasite T. gondii as a new trigger of prostatic inflammation, which we used to develop a novel mouse model of prostatic inflammation. This is the first report that T. gondii chronically encysts and induces chronic inflammation within the prostate of any species. Furthermore, T. gondii-induced prostatic inflammation persists and progresses without genetic manipulation in mice, offering a powerful new mouse model for the study of chronic prostatic inflammation and microglandular hyperplasia. © 2017 Wiley Periodicals, Inc.

  6. Low titer of antibody against Toxoplasma gondii may be related to resistant to cancer

    OpenAIRE

    Maryam Sharafi Seyedeh; Salehi Nahid; Mortazavi Nahid; Danesh Pour Shima; Yousefi Morteza; Yousofi Darani Hossein

    2015-01-01

    Context: Toxoplasma gondii is a protozoan parasite with a world-wide distribution. However, the majority of infected cases remain symptomless. There are raising scientific evidences indicating that parasitic infections induce antitumor activity against certain types of cancers. The inhibitory effect of T. gondii on cancer growth has also been shown in cell culture and mouse model. Aims: Considering the anti-tumor effect of this parasite, in this study the relationship between low titer of ...

  7. Seropositivity and risk factors associated with Toxoplasma gondii infection in wild birds from Spain.

    Directory of Open Access Journals (Sweden)

    Oscar Cabezón

    Full Text Available Toxoplasma gondii is a zoonotic intracellular protozoan parasite of worldwide distribution that infects many species of warm-blooded animals, including birds. To date, there is scant information about the seropositivity of T. gondii and the risk factors associated with T. gondii infection in wild bird populations. In the present study, T. gondii infection was evaluated on sera obtained from 1079 wild birds belonging to 56 species (including Falconiformes (n=610, Strigiformes (n=260, Ciconiiformes (n=156, Gruiformes (n=21, and other orders (n=32, from different areas of Spain. Antibodies to T. gondii (modified agglutination test, MAT titer ≥1:25 were found in 282 (26.1%, IC(95%:23.5-28.7 of the 1079 birds. This study constitute the first extensive survey in wild birds species in Spain and reports for the first time T. gondii antibodies in the griffon vulture (Gyps fulvus, short-toed snake-eagle (Circaetus gallicus, Bonelli's eagle (Aquila fasciata, golden eagle (Aquila chrysaetos, bearded vulture (Gypaetus barbatus, osprey (Pandion haliaetus, Montagu's harrier (Circus pygargus, Western marsh-harrier (Circus aeruginosus, peregrine falcon (Falco peregrinus, long-eared owl (Asio otus, common scops owl (Otus scops, Eurasian spoonbill (Platalea leucorodia, white stork (Ciconia ciconia, grey heron (Ardea cinerea, common moorhen (Gallinula chloropus; in the International Union for Conservation of Nature (IUCN "vulnerable" Spanish imperial eagle (Aquila adalberti, lesser kestrel (Falco naumanni and great bustard (Otis tarda; and in the IUCN "near threatened" red kite (Milvus milvus. The highest seropositivity by species was observed in the Eurasian eagle owl (Bubo bubo (68.1%, 98 of 144. The main risk factors associated with T. gondii seropositivity in wild birds were age and diet, with the highest exposure in older animals and in carnivorous wild birds. The results showed that T. gondii infection is widespread and can be at a high level in many wild

  8. Chronic Toxoplasma gondii in Nurr1-Null Heterozygous Mice Exacerbates Elevated Open Field Activity

    OpenAIRE

    Eells, Jeffrey B.; Varela-Stokes, Andrea; Guo-Ross, Shirley X.; Kummari, Evangel; Smith, Holly M.; Cox, Erin; Lindsay, David S.

    2015-01-01

    Latent infection with Toxoplasma gondii is common in humans (approximately 30% of the global population) and is a significant risk factor for schizophrenia. Since prevalence of T. gondii infection is far greater than prevalence of schizophrenia (0.5-1%), genetic risk factors are likely also necessary to contribute to schizophrenia. To test this concept in an animal model, Nurr1-null heterozygous (+/-) mice and wild-type (+/+) mice were evaluate using an emergence test, activity in an open fie...

  9. Behavioral changes in Rattus norvegicus coinfected by Toxocara canis and Toxoplasma gondii

    Directory of Open Access Journals (Sweden)

    Maisa Leite de Queiroz

    2013-02-01

    Full Text Available Using an elevated plus maze apparatus and an activity cage, behavioral changes in Rattus norvegicus concomitantly infected by Toxocara canis and Toxoplasma gondii were studied, during a period of 120 days. Rats infected by Toxocara canis or Toxoplasma gondii showed significant behavioral changes; however, in the group coinfected by both parasites a behavioral pattern similar to that found in the group not infected was observed thirty days after infection, suggesting the occurrence of modulation in the behavioral response.

  10. Loop-mediated isothermal amplification (LAMP): Early detection of Toxoplasma gondii infection in mice

    OpenAIRE

    Kong, Qing-Ming; Lu, Shao-Hong; Tong, Qun-Bo; Lou, Di; Chen, Rui; Zheng, Bin; Kumagai, Takashi; Wen, Li-Yong; Ohta, Nobuo; Zhou, Xiao-Nong

    2012-01-01

    Abstract Background Toxoplasmosis is a widespread zoonotic parasitic disease that occurs in both animals and humans. Traditional molecular assays are often difficult to perform, especially for the early diagnosis of Toxoplasma gondii infections. Here, we established a novel loop-mediated isothermal amplification targeting the 529 bp repeat element (529 bp-LAMP) to detect T. gondii DNA in blood samples of experimental mice infected with tachyzoites of the RH strain. Findings The assay was perf...

  11. THE ROLE OF HORMONES ON Toxoplasma gondii INFECTION: A SYSTEMATIC REVIEW

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    María De La Luz Galván-Ramírez

    2014-10-01

    Full Text Available Background: Toxoplasma gondii is the causal agent of toxoplasmosis in which one third of the world’s population has been infected. In pregnant women, it may cause abortion and severe damage to the fetal central nervous system. During pregnancy, the prevalence of toxoplasmosis increases throughout the second and third quarter of gestation, simultaneously progesterone and 17β-estradiol also increase. Thus, it has been suggested that these hormones can aggravate or reduce parasite reproduction. The aim of this study was reviewing the relationship between hormones and infection caused by T. gondii in several experimental animal models and humans, focused mainly on: a congenital transmission, b parasite reproduction, c strain virulence, d levels of hormone in host induced by T. gondii infection and e participation of hormone receptors in Toxoplasma gondii infection.Are the hormones specific modulators of T. gondii infection?A systematic review methodology was used to consult several databases (Pub Med, Lilacs, Medline, Science direct, Scielo, Ebsco, Sprinker, Wiley and Google Scholar dated from September, 2013 to March, 2014. Results: 30 studies were included; eight studies in humans and 22 in animals and cell cultures. In the human studies, the most studied hormones were testosterone, progesterone, prolactin and 17-ß estradiol. Type I (RH and BK and Type II (Prugniaud, SC, ME49,T45, P78 and T38 were the most frequent experimental strains. Conclusions: Thirty-five years have passed since the first studies regarding Toxoplasma gondii infection and its relationship with hormones. This systematic review suggests that hormones modulate Toxoplasma gondii infection in different animal models. However, given that data were not comparable, further studies are required to determine the mechanism of hormone action in the Toxoplasma gondii infectious process.

  12. Screening and identification of novel B cell epitopes of Toxoplasma gondii SAG1

    OpenAIRE

    Wang, Yanhua; Wang, Guangxiang; Zhang, Delin; Yin, Hong; Wang, Meng

    2013-01-01

    Background The identification of protein epitopes is useful for diagnostic purposes and for the development of peptide vaccines. In this study, the epitopes of Toxoplasma gondii SAG1 were identified using synthetic peptide techniques with the aid of bioinformatics. Findings Eleven peptides derived from T. gondii SAG1 were assessed by ELISA using pig sera from different time points after infection. Four (PS4, PS6, PS10 and PS11), out of the eleven peptides tested were recognized by all sera. T...

  13. Detection of Toxoplasma gondii DNA in horse meat from supermarkets in France and performance evaluation of two serological tests

    OpenAIRE

    Aroussi Abdelkrim; Vignoles Philippe; Dalmay François; Wimel Laurence; Dardé Marie-Laure; Mercier Aurélien; Ajzenberg Daniel

    2015-01-01

    In France, some cases of severe toxoplasmosis have been linked to the consumption of horse meat that had been imported from the American continent where atypical strains of Toxoplasma gondii are more common than in Europe. Many seroprevalence studies are presented in the literature but risk assessment of T. gondii infection after horse meat consumption is not possible in the absence of validated serological tests and the unknown correlation between detection of antibodies against T. gondii an...

  14. Incidence of adult brain cancers is higher in countries where the protozoan parasite Toxoplasma gondii is common

    OpenAIRE

    Thomas, Frédéric; Lafferty, Kevin D.; Brodeur, Jacques; Elguero, Eric; Gauthier-Clerc, Michel; Missé, Dorothée

    2011-01-01

    We explored associations between the common protozoan parasite Toxoplasma gondii and brain cancers in human populations. We predicted that T. gondii could increase the risk of brain cancer because it is a long-lived parasite that encysts in the brain, where it provokes inflammation and inhibits apoptosis. We used a medical geography approach based on the national incidence of brain cancers and seroprevalence of T. gondii. We corrected reports of incidence for national gross domestic product b...

  15. Seroprevalence of Toxoplasma gondii and Neospora caninum in red deer from Central Italy

    Directory of Open Access Journals (Sweden)

    Guido Rocchigiani

    2016-09-01

    Full Text Available Neospora caninum and Toxoplasma gondii are cosmopolite protozoan parasites impacting on human and animal health. In particular, T. gondii commonly infects human beings and all warm-blooded animals, while N. caninum is responsible for bovine abortion and neuromuscular disease in dogs. The aim of the presented survey was to evaluate the occurrence and prevalence of these parasites in the most numerous Italian red deer population. The sera of 60 red deer ( Cervus elaphus inhabiting Central Italy (43°56’N 10°55’E and killed by selective hunting were examined using an indirect fluorescent antibody test (IFAT for both N. caninum and T. gondii antibodies. White blood cells (buffy coat were also checked by PCR and T. gondii DNA was genotyped. Thirteen out of 60 sera (22% scored positive for Toxoplasma, 17 samples (28% were Neospora positive. Coinfection was recorded in 5 cases (8%. T. gondii (genotype II and N. caninum DNA was detected in one and 3 samples of buffy coat, respectively. The presented study is the first to examine the occurrence of these parasites in the most numerous red deer Italian population, confirming this animal species as carrier of the investigated pathogens. These animals spread near human settlements, co-inhabiting with final hosts of [i]T. gondii[/i] and N. caninum and could contribute to their transmission to domestic ruminants and humans. In particular, the seroprevalence value for N. caninum was the highest among European records.

  16. Toxoplasma gondii transmission by artificial insemination in sheep with experimentally contaminated frozen semen.

    Science.gov (United States)

    Consalter, Angélica; Silva, Andressa F; Frazão-Teixeira, Edwards; Matos, Luis F; de Oliveira, Francisco C R; Leite, Juliana S; Silva, Franciele B F; Ferreira, Ana M R

    2017-03-01

    Toxoplasma gondii is a parasite considered one of the major causes of reproductive problems in sheep. Furthermore, the presence of the agent in ram semen urges the possibility of sexual transmission in this species. The aim of this study was to evaluate if ram's frozen semen spiked with T. gondii tachyzoites would be able to cause infection in sheep by laparoscopic artificial insemination (AI). Nine ewes tested seronegative to anti-T. gondii antibodies by the modified agglutination test (MAT) were superovulated and inseminated to collect embryos. Animals were divided into two groups: G1 (n = 5), ewes inseminated with semen containing 4 × 10 7 tachyzoites; and G2 (n = 4), ewes inseminated with tachyzoite-free semen (control group). To confirm infection, ewe's blood samples were collected on days -14, -7, 0, 7, 14, 21, 28, 35, 49 and 57 after AI for analysis by MAT and PCR. Tissue samples of these ewes were also collected for histopathology and immunohistochemistry (IHC). Seven days after AI, all ewes of group G1 had specific antibodies to T. gondii, while those of G2 were negative. Toxoplasma gondii DNA was detected in the blood of one ewe and parasites were observed in tissues of all five animals inseminated with contaminated semen, indicating that semen freezing protocol does not affect T. gondii transmission by artificial insemination in sheep. Copyright © 2016 Elsevier Inc. All rights reserved.

  17. A qualitative assessment of Toxoplasma gondii risk in ready-to-eat smallgoods processing.

    Science.gov (United States)

    Mie, Tanya; Pointon, Andrew M; Hamilton, David R; Kiermeier, Andreas

    2008-07-01

    Toxoplasma gondii is one of the most common parasitic infections of humans and other warm-blooded animals. In most adults, it does not cause serious illness, but severe disease may result from infection in fetuses and immunocompromised people. Consumption of raw or undercooked meats has consistently been identified as an important source of exposure to T. gondii. Several studies indicate the potential failure to inactivate T. gondii in the processes of cured meat products, This article presents a qualitative risk-based assessment of the processing of ready-to-eat smallgoods, which include cooked or uncooked fermented meat, pâté, dried meat, slow cured meat, luncheon meat, and cooked muscle meat including ham and roast beef. The raw meat ingredients are rated with respect to their likelihood of containing T. gondii cysts and an adjustment is made based on whether all the meat from a particular source is frozen. Next, the effectiveness of common processing steps to inactivate T. gondii cysts is assessed, including addition of spices, nitrates, nitrites and salt, use of fermentation, smoking and heat treatment, and the time and temperature during maturation. It is concluded that processing steps that may be effective in the inactivation of T. gondii cysts include freezing, heat treatment, and cooking, and the interaction between salt concentration, maturation time, and temperature. The assessment is illustrated using a Microsoft Excel-based software tool that was developed to facilitate the easy assessment of four hypothetical smallgoods products.

  18. Computational Prediction of MicroRNAs from Toxoplasma gondii Potentially Regulating the Hosts’ Gene Expression

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    Müşerref Duygu Saçar

    2014-10-01

    Full Text Available MicroRNAs (miRNAs were discovered two decades ago, yet there is still a great need for further studies elucidating their genesis and targeting in different phyla. Since experimental discovery and validation of miRNAs is difficult, computational predictions are indispensable and today most computational approaches employ machine learning. Toxoplasma gondii, a parasite residing within the cells of its hosts like human, uses miRNAs for its post-transcriptional gene regulation. It may also regulate its hosts’ gene expression, which has been shown in brain cancer. Since previous studies have shown that overexpressed miRNAs within the host are causal for disease onset, we hypothesized that T. gondii could export miRNAs into its host cell. We computationally predicted all hairpins from the genome of T. gondii and used mouse and human models to filter possible candidates. These were then further compared to known miRNAs in human and rodents and their expression was examined for T. gondii grown in mouse and human hosts, respectively. We found that among the millions of potential hairpins in T. gondii, only a few thousand pass filtering using a human or mouse model and that even fewer of those are expressed. Since they are expressed and differentially expressed in rodents and human, we suggest that there is a chance that T. gondii may export miRNAs into its hosts for direct regulation.

  19. Serological evidence of Toxoplasma gondii infection in captive marine mammals in Mexico.

    Science.gov (United States)

    Alvarado-Esquivel, C; Sánchez-Okrucky, R; Dubey, J P

    2012-03-23

    Toxoplasma gondii infection in marine mammals is important because they are considered as a sentinel for contamination of seas with T. gondii oocysts, and toxoplasmosis causes mortality in these animals, particularly sea otters. Serological evidence of T. gondii infection was determined in 75 captive marine mammals from four facilities in southern and central geographical regions in Mexico using the modified agglutination test (MAT). Antibodies (MAT, 1:25 or higher) to T. gondii were found in 55 (87.3%) of 63 Atlantic bottlenose dolphins (Tursiops truncatus truncatus), 3 of 3 Pacific bottlenose dolphins (Tursiops truncatus gillii), 2 of 4 California sea lions (Zalophus californianus), but not in 3 West Indian manatees (Trichechus manatus), and 2 Patagonian sea lions (Otaria flavescens). Seropositive marine mammals were found in all 4 (100%) facilities sampled. All marine mammals were healthy and there has not been any case of clinical toxoplasmosis in the facilities sampled for at least the last 15 years. The seroprevalence of T. gondii infection in marine mammals of the same species did not vary significantly with respect to sex and age. This is the first report on the detection of antibodies to T. gondii in marine mammals in Mexico. Published by Elsevier B.V.

  20. Toxoplasmosis in dogs: first report of Toxoplasma gondii infection in any animal species in Angola.

    Science.gov (United States)

    Lopes, Ana Patrícia; Granada, Sara; Oliveira, Ana Cristina; Brancal, Hugo; Dubey, Jitender P; Cardoso, Luís; Vilhena, Hugo

    2014-10-01

    Despite the worldwide importance of zoonotic parasite Toxoplasma gondii, nothing is known of toxoplasmosis in animals in Angola. The present study aimed at estimating the seroprevalence and also at assessing correlates of T. gondii infection in pet dogs from Luanda, Angola. Dogs (n = 103) brought to a veterinary clinic in the city of Luanda were investigated. Serum samples were tested for antibodies to T. gondii with a modified agglutination test (MAT) commercial kit, at serial dilutions of 1∶20 to 1∶160. In accordance with the established cutoff value (MAT ≧20), 16 dogs [15·5%; 95% confidence interval (CI): 9·2-24·0%] had antibodies to T. gondii: 10 had a titer of 20, two had a titer of 40, and four had a titer of 80. Age (≧12 months) was found to be a risk factor for infection [odds ratio (OR) = 9·23; 95% CI: 1·16-73·27). For each 1-year increase in age, the risk of a dog being found seropositive significantly increased by an OR of 1·18 (95% CI: 1·02-1·36). The present study, which represents the first serological survey of T. gondii in any animal species from Angola, reveals a 15·5% seroprevalence of infection in pet dogs in Luanda. Further studies are needed to better understand the epidemiology of zoonotic T. gondii infection in Luanda and also in Angola.

  1. Prevalence of Toxoplasma gondii in Chicken samples from delta of Egypt using ELISA, histopathology and immunohistochemistry.

    Science.gov (United States)

    Ibrahim, Hany M; Abdel-Ghaffar, Fathy; Osman, Gamalat Y; El-Shourbagy, Safinaz H; Nishikawa, Yoshifumi; Khattab, Reham A

    2016-06-01

    Estimates of the zoonotic diseases are helpful for monitoring and improving public health. Laboratory-based surveillance provides crucial information for assessing zoonotic disease trends and developments. Toxoplasmosis is considered as a zoonotic disease and has both medical and veterinary importance since it leads to abortion in humans and several animal species. In view of the worldwide importance of T. gondii, this study aimed to estimate the prevalence of T. gondii in chickens from the Delta of Egypt. A total of 304 blood and brain samples were collected from Egyptian chickens from Gharbiya, Qalyoubiya, Minufiya, Beheira, Kafr EL-Shaykh and Dakahlia Provinces. In order to determine the serological and histopathological prevalence of T. gondii, the samples were examined by ELISA, histopathology and immunohistochemistry (IHC). The prevalence of T. gondii was 11.18, 6.91, 6.91 % by ELISA, histopathology and IHC, respectively. Statistically significant differences in the prevalence of T. gondii were observed on the basis of season, sex and habitat. These data provide valuable information regarding the epidemiology of T. gondii infections in Egyptian chickens, which can be employed in developing efficient strategies for disease management and control.

  2. Stillbirth history and Toxoplasma gondii infection in women attending public health centers in a northern Mexican City

    Science.gov (United States)

    Alvarado-Esquivel, C.; Pacheco-Vega, S. J.; Salcedo-Jaquez, M.; Sánchez-Anguiano, L. F.; Hernández-Tinoco, J.; Rábago-Sánchez, E.; Centeno-Tinoco, M. M.; Flores-Garcia, I. D.; Ramos-Nevarez, A.; Cerrillo-Soto, S. M.; Guido-Arreola, C. A.; Beristain-García, I.; Liesenfeld, O.; Berumen-Segovia, L. O.; Saenz-Soto, L.; Sifuentes-Álvarez, A.

    2015-01-01

    Through a cross-sectional study design, 150 women attending public health centers with a history of stillbirths were examined for anti-Toxoplasma gondii IgG and IgM antibodies in Durango City, Mexico. Bivariate and multivariate analyses were used to assess the association of T. gondii seropositivity with the characteristics of the women with stillbirth history. Of the 150 women (mean age: 32.09 ± 9.16 years) studied, 14 (9.3%) had anti-T. gondii IgG antibodies and six (42.9%) of them were also positive for anti-T. gondii IgM antibodies. Multivariate analysis showed that T. gondii seropositivity was associated with high frequency (4–7 days a week) of eating meat (OR = 5.52; 95% CI: 1.48–20.59; P = 0.01), history of lymphadenopathy (OR = 4.52; 95% CI: 1.14–17.82; P = 0.03), and history of surgery (OR = 8.68; 95% CI: 1.04–72.15; P = 0.04). This is the first study on the seroepidemiology of T. gondii infection in women with a history of stillbirths in Mexico. The association of T. gondii exposure with a history of surgery warrants for further research. Risk factors for T. gondii infection found in the present survey may help to design optimal educational programs to avoid T. gondii infection. PMID:26185685

  3. Prevalence of Toxoplasma gondii antibodies in domestic donkeys (Equus asinus) in Durango, Mexico slaughtered for human consumption.

    Science.gov (United States)

    Alvarado-Esquivel, Cosme; Alvarado-Esquivel, Domingo; Dubey, Jitender P

    2015-01-17

    Nothing is known about Toxoplasma gondii prevalence in donkeys in Mexico. Meat from donkey is consumed by humans in Mexico and also exported to other countries. We sought to determine the presence of antibodies against T. gondii in 239 domestic donkeys (Equus asinus) for slaughter in Durango, Mexico using the modified agglutination test (MAT). Donkeys were sampled in four premises (trade centers) where donkeys were gather for shipment to abattoirs in other Mexican states. Antibodies to T. gondii were found in 26 (10.9%) of 239 donkeys, with MAT titers of 1:25 in 7, 1:50 in 11, 1:100 in 6, and 1:200 in 2. Seropositive donkeys were found in three (75%) of the four gathering premises studied. Seroprevalence in donkeys varied from 0% to 23.1% among gathering premises. The seroprevalence of T. gondii infection was comparable among donkeys regardless their age, sex or health status. Seropositivity to T. gondii was found in donkeys between 1 to 12 years old. Multivariate analysis showed that seropositivity to T. gondii was associated with the gathering premises (OR = 1.58; 95% CI: 1.11-2.24; P = 0.009). This is the first report of T. gondii infection in donkeys in Mexico. Results indicate that consumption of undercooked or raw meat from T. gondii-infected donkeys is potentially a source of T. gondii infection for humans.

  4. Anti-Toxoplasma gondii secretory IgA in tears of patients with ocular toxoplasmosis: immunodiagnostic validation by ELISA

    OpenAIRE

    Lynch,Maria Isabel; Malagueño,Elizabeth; Lynch,Luiz Felipe; Ferreira,Silvana; Stheling,Raphael; Oréfice,Fernando

    2009-01-01

    Toxoplasma gondii causes posterior uveitis and the specific diagnosis is based on clinical criteria. The presence of anti-T. gondii secretory IgA (sIgA) antibodies in patients' tears has been reported and an association was found between ocular toxoplasmosis and the anti-T. gondii sIgA isotype in Brazilian patients. The purpose of this study was to provide an objective validation of the published ELISA test for determining the presence of anti-T. gondii sIgA in the tears of individuals with o...

  5. From Drosophila to humans: Reflections on the roles of the prolyl-isomerases and chaperones, cyclophilins, in cell function and disease

    Science.gov (United States)

    Ferreira, Paulo A.; Orry, Andrew

    2013-01-01

    Despite remarkable advances in human genetics and other genetic model systems, the fruit fly, Drosophila melanogaster, remains a powerful experimental tool to probe with ease the inner workings of a myriad of biological and pathological processes, even when evolutionary forces impart apparent divergences to some of such processes. The understanding of such evolutionary differences provides mechanistic insights into genotype-phenotype correlations underpinning biological processes across metazoans. The pioneering work developed by the William Pak laboratory for the past four decades, and the work of others, epitomize the notion of how the Drosophila system breaks new fertile ground or complements research fields of high scientific and medical relevance. Among the three major genetic complementation groups produced by the Pak's laboratory and impairing distinct facets of photoreceptor neuronal function, the nina group (ninaA….J) selectively affects the biogenesis of G protein-coupled receptors (GPCR) mediating the photoconversion and transduction of light-stimuli. Among the nina genes identified, ninaA arguably assumes heightened significance for several reasons. First, it presents unique physiological selectivity toward the biogenesis of a subset of GPCRs, a standalone biological manifestation yet to be discerned for most mammalian homologues of NinaA. Second, NinaA belongs to a family of proteins, immunophilins, which are the primary targets for immunosuppressive drugs at the therapeutic forefront of a multitude of medical conditions. Third, NinaA closest homologue, cyclophilin-B (CyPB/PPIB), is an immunophilin whose loss-of-function was found recently to cause osteogenesis imperfecta in the human. This report highlights advances made by studies on some members of immunophilins, the cyclophilins. Finally, it re-examines critically data and dogmas derived from past and recent genetic, structural, biological and pathological studies on NinaA and few other

  6. Molecular dynamics simulations of site point mutations in the TPR domain of cyclophilin 40 identify conformational states with distinct dynamic and enzymatic properties

    Science.gov (United States)

    Gur, Mert; Blackburn, Elizabeth A.; Ning, Jia; Narayan, Vikram; Ball, Kathryn L.; Walkinshaw, Malcolm D.; Erman, Burak

    2018-04-01

    Cyclophilin 40 (Cyp40) is a member of the immunophilin family that acts as a peptidyl-prolyl-isomerase enzyme and binds to the heat shock protein 90 (Hsp90). Its structure comprises an N-terminal cyclophilin domain and a C-terminal tetratricopeptide (TPR) domain. Cyp40 is overexpressed in prostate cancer and certain T-cell lymphomas. The groove for Hsp90 binding on the TPR domain includes residues Lys227 and Lys308, referred to as the carboxylate clamp, and is essential for Cyp40-Hsp90 binding. In this study, the effect of two mutations, K227A and K308A, and their combinative mutant was investigated by performing a total of 5.76 μs of all-atom molecular dynamics (MD) simulations in explicit solvent. All simulations, except the K308A mutant, were found to adopt two distinct (extended or compact) conformers defined by different cyclophilin-TPR interdomain distances. The K308A mutant was only observed in the extended form which is observed in the Cyp40 X-ray structure. The wild-type, K227A, and combined mutant also showed bimodal distributions. The experimental melting temperature, Tm, values of the mutants correlate with the degree of compactness with the K308A extended mutant having a marginally lower melting temperature. Another novel measure of compactness determined from the MD data, the "coordination shell volume," also shows a direct correlation with Tm. In addition, the MD simulations show an allosteric effect with the mutations in the remote TPR domain having a pronounced effect on the molecular motions of the enzymatic cyclophilin domain which helps rationalise the experimentally observed increase in enzyme activity measured for all three mutations.

  7. Evasion of Human Neutrophil-Mediated Host Defense during Toxoplasma gondii Infection.

    Science.gov (United States)

    Lima, Tatiane S; Gov, Lanny; Lodoen, Melissa B

    2018-02-13

    Neutrophils are a major player in host immunity to infection; however, the mechanisms by which human neutrophils respond to the intracellular protozoan parasite Toxoplasma gondii are still poorly understood. In the current study, we found that, whereas primary human monocytes produced interleukin-1beta (IL-1β) in response to T. gondii infection, human neutrophils from the same blood donors did not. Moreover, T. gondii inhibited lipopolysaccharide (LPS)-induced IL-1β synthesis in human peripheral blood neutrophils. IL-1β suppression required active parasite invasion, since heat-killed or mycalolide B-treated parasites did not inhibit IL-1β release. By investigating the mechanisms involved in this process, we found that T. gondii infection of neutrophils treated with LPS resulted in reduced transcript levels of IL-1β and NLRP3 and reduced protein levels of pro-IL-1β, mature IL-1β, and the inflammasome sensor NLRP3. In T. gondii -infected neutrophils stimulated with LPS, the levels of MyD88, TRAF6, IKKα, IKKβ, and phosphorylated IKKα/β were not affected. However, LPS-induced IκBα degradation and p65 phosphorylation were reduced in T. gondii- infected neutrophils, and degradation of IκBα was reversed by treatment with the proteasome inhibitor MG-132. Finally, we observed that T. gondii inhibited the cleavage and activity of caspase-1 in human neutrophils. These results indicate that T. gondii suppression of IL-1β involves a two-pronged strategy whereby T. gondii inhibits both NF-κB signaling and activation of the NLRP3 inflammasome. These findings represent a novel mechanism of T. gondii evasion of human neutrophil-mediated host defense by targeting the production of IL-1β. IMPORTANCE Toxoplasma gondii is an obligate intracellular parasite that infects approximately one-third of humans worldwide and can invade virtually any nucleated cell in the human body. Although it is well documented that neutrophils infiltrate the site of acute T. gondii

  8. Hepatitis C virus NS5A protein is a substrate for the peptidyl-prolyl cis/trans isomerase activity of cyclophilins A and B.

    Science.gov (United States)

    Hanoulle, Xavier; Badillo, Aurélie; Wieruszeski, Jean-Michel; Verdegem, Dries; Landrieu, Isabelle; Bartenschlager, Ralf; Penin, François; Lippens, Guy

    2009-05-15

    We report here a biochemical and structural characterization of domain 2 of the nonstructural 5A protein (NS5A) from the JFH1 Hepatitis C virus strain and its interactions with cyclophilins A and B (CypA and CypB). Gel filtration chromatography, circular dichroism spectroscopy, and finally NMR spectroscopy all indicate the natively unfolded nature of this NS5A-D2 domain. Because mutations in this domain have been linked to cyclosporin A resistance, we used NMR spectroscopy to investigate potential interactions between NS5A-D2 and cellular CypA and CypB. We observed a direct molecular interaction between NS5A-D2 and both cyclophilins. The interaction surface on the cyclophilins corresponds to their active site, whereas on NS5A-D2, it proved to be distributed over the many proline residues of the domain. NMR heteronuclear exchange spectroscopy yielded direct evidence that many proline residues in NS5A-D2 form a valid substrate for the enzymatic peptidyl-prolyl cis/trans isomerase (PPIase) activity of CypA and CypB.

  9. Serological study for antibodies to toxoplasma gondii in Jakarta, Indonesia.

    Science.gov (United States)

    Gandahusada, S

    1978-09-01

    A total of 280 sera from medical students, laboratory personnel from the University of Indonesia and other persons living in Jakarta were tested for antibodies to Toxoplasma gondii by the indirect hemagglutination test. Antibody titers equal to or greater than 1:256 were considered positive and were detected in 35 or 12.5% of the persons tested. The sero-positivity rates were not significant between 178 males (13.5%) and 102 females (10.8%) but were significantly different between persons of Indonesian ancestry (14.3%) and those of Chinese ancestry (2.3%). No correlation could be found between ownership of domestic cats and eating habits and positive titers.

  10. Toxoplasma gondii, Neospora caninum, Sarcocystis neurona, and Sarcocystis canis-like infections in marine mammals

    Science.gov (United States)

    Dubey, J.P.; Zarnke, R.; Thomas, N.J.; Wong, S.K.; Vanbonn, W.; Briggs, M.; Davis, J.W.; Ewing, R.; Mense, M.; Kwok, O.C.H.; Romand, S.; Thulliez, P.

    2003-01-01

    Toxoplasma gondii, Neospora caninum, Sarcocystis neurona, and S. canis are related protozoans that can cause mortality in many species of domestic and wild animals. Recently, T. gondii and S. neurona were recognized to cause encephalitis in marine mammals. As yet, there is no report of natural exposure of N. caninum in marine mammals. In the present study, antibodies to T. gondii and N. caninum were assayed in sera of several species of marine mammals. For T. gondii, sera were diluted 1:25, 1:50, and 1:500 and assayed in the T. gondii modified agglutination test (MAT). Antibodies (MAT a?Y1:25) to T. gondii were found in 89 of 115 (77%) dead, and 18 of 30 (60%) apparently healthy sea otters (Enhydra lutris), 51 of 311 (16%) Pacific harbor seals (Phoca vitulina), 19 of 45 (42%) sea lions (Zalophus californianus), 5 of 32 (16%) ringed seals (Phoca hispida), 4 of 8 (50%) bearded seals (Erignathus barbatus), 1 of 9 (11.1%) spotted seals (Phoca largha), 138 of 141 (98%) Atlantic bottlenose dolphins (Tursiops truncatus), and 3 of 53 (6%) walruses (Odobenus rosmarus). For N. caninum, sera were diluted 1:40, 1:80, 1:160, and 1:320 and examined with the Neospora agglutination test (NAT) using mouse-derived tachyzoites. NAT antibodies were found in 3 of 53 (6%) walruses, 28 of 145 (19%) sea otters, 11 of 311 (3.5%) harbor seals, 1 of 27 (3.7%) sea lions, 4 of 32 (12.5%) ringed seals, 1 of 8 (12.5%) bearded seals, and 43 of 47 (91%) bottlenose dolphins. To our knowledge, this is the first report of N. caninum antibodies in any marine mammal, and the first report of T. gondii antibodies in walruses and in ringed, bearded, spotted, and ribbon seals. Current information on T. gondii-like and Sarcocystis-like infections in marine mammals is reviewed. New cases of clinical S. canis and T. gondii infections are also reported in sea lions, and T. gondii infection in an Antillean manatee (Trichechus manatus manatus).

  11. Antibody Detection, Isolation, Genotyping, and Virulence of Toxoplasma gondii in Captive Felids from China

    Directory of Open Access Journals (Sweden)

    Yu-Rong Yang

    2017-07-01

    Full Text Available The felids are the only definitive hosts of Toxoplasma gondii, which could excrete oocysts into the environment and provide an infection source for toxoplasmosis in various warm-blooded animal species, particularly the captive felids that live close to human communities. The infection rate of the captive felids is a perfect standard in detecting the presence of Toxoplasma gondii oocysts in the environment. In this study, sera or tissue samples from zoo (1 young tiger, 2 adult tigers, 6 young lions, farm (10 masked palm civets, and pet hospital (28 cats from Henan Province (China were collected. The sera (n = 47 were tested for immunoglobulin G (IgG antibodies against T. gondii by using modified agglutination test (MAT, whereas the hearts tissue (n = 40 were bioassayed in mice to isolate T. gondii strains. The genotype was distinguished by using PCR-RFLP of 10 loci (SAG1, SAG2, SAG3, GRA6, BTUB, L358, c22-8, PK1, c29-2, and Apico. The detection rate for the T. gondii antibody in captive felids was 21.3% (10/47. One viable T. gondii strain (TgCatCHn4 was obtained from a cat heart tissue, and its genotype was ToxoDB#9. The oocysts of ToxoDB#9 were collected from a T. gondii-free cat. The virulence of TgCatCHn4 was low and no cysts were detected in the brain of mice at 60 days post-inoculation. The finding of the present study suggested a widespread exposure of T. gondii for felids in Henan Province of central China, particularly those from the zoological gardens and homes. ToxoDB#9 was the predominant strain in China. Preventive measures against T. gondii oocyst contamination of various components of the environment should thus be implemented, including providing pre-frozen meat, well-cooked cat food, cleaned fruits and vegetables, monitoring birds and rodents, inactive T. gondii oocysts in felids feces, and proper hygiene.

  12. The crystal structure of Toxoplasma gondii pyruvate kinase 1.

    Directory of Open Access Journals (Sweden)

    Rebecca Bakszt

    2010-09-01

    Full Text Available Pyruvate kinase (PK, which catalyzes the final step in glycolysis converting phosphoenolpyruvate to pyruvate, is a central metabolic regulator in most organisms. Consequently PK represents an attractive therapeutic target in cancer and human pathogens, like Apicomplexans. The phylum Aplicomplexa, a group of exclusively parasitic organisms, includes the genera Plasmodium, Cryptosporidium and Toxoplasma, the etiological agents of malaria, cryptosporidiosis and toxoplasmosis respectively. Toxoplasma gondii infection causes a mild illness and is a very common infection affecting nearly one third of the world's population.We have determined the crystal structure of the PK1 enzyme from T. gondii, with the B domain in the open and closed conformations. We have also characterized its enzymatic activity and confirmed glucose-6-phosphate as its allosteric activator. This is the first description of a PK enzyme in a closed inactive conformation without any bound substrate. Comparison of the two tetrameric TgPK1 structures indicates a reorientation of the monomers with a concomitant change in the buried surface among adjacent monomers. The change in the buried surface was associated with significant B domain movements in one of the interacting monomers.We hypothesize that a loop in the interface between the A and B domains plays an important role linking the position of the B domain to the buried surface among monomers through two α-helices. The proposed model links the catalytic cycle of the enzyme with its domain movements and highlights the contribution of the interface between adjacent subunits. In addition, an unusual ordered conformation was observed in one of the allosteric binding domains and it is related to a specific apicomplexan insertion. The sequence and structural particularity would explain the atypical activation by a mono-phosphorylated sugar. The sum of peculiarities raises this enzyme as an emerging target for drug discovery.

  13. The crystal structure of Toxoplasma gondii pyruvate kinase 1.

    Science.gov (United States)

    Bakszt, Rebecca; Wernimont, Amy; Allali-Hassani, Abdellah; Mok, Man Wai; Hills, Tanya; Hui, Raymond; Pizarro, Juan C

    2010-09-14

    Pyruvate kinase (PK), which catalyzes the final step in glycolysis converting phosphoenolpyruvate to pyruvate, is a central metabolic regulator in most organisms. Consequently PK represents an attractive therapeutic target in cancer and human pathogens, like Apicomplexans. The phylum Aplicomplexa, a group of exclusively parasitic organisms, includes the genera Plasmodium, Cryptosporidium and Toxoplasma, the etiological agents of malaria, cryptosporidiosis and toxoplasmosis respectively. Toxoplasma gondii infection causes a mild illness and is a very common infection affecting nearly one third of the world's population. We have determined the crystal structure of the PK1 enzyme from T. gondii, with the B domain in the open and closed conformations. We have also characterized its enzymatic activity and confirmed glucose-6-phosphate as its allosteric activator. This is the first description of a PK enzyme in a closed inactive conformation without any bound substrate. Comparison of the two tetrameric TgPK1 structures indicates a reorientation of the monomers with a concomitant change in the buried surface among adjacent monomers. The change in the buried surface was associated with significant B domain movements in one of the interacting monomers. We hypothesize that a loop in the interface between the A and B domains plays an important role linking the position of the B domain to the buried surface among monomers through two α-helices. The proposed model links the catalytic cycle of the enzyme with its domain movements and highlights the contribution of the interface between adjacent subunits. In addition, an unusual ordered conformation was observed in one of the allosteric binding domains and it is related to a specific apicomplexan insertion. The sequence and structural particularity would explain the atypical activation by a mono-phosphorylated sugar. The sum of peculiarities raises this enzyme as an emerging target for drug discovery.

  14. The Crystal Structure of Toxoplasma gondii Pyruvate Kinase 1

    Energy Technology Data Exchange (ETDEWEB)

    Bakszt, R.; Wernimont, A; Allali-Hassani, A; Mok, M; Hills, T; Hui, R; Pizarro, J

    2010-01-01

    Pyruvate kinase (PK), which catalyzes the final step in glycolysis converting phosphoenolpyruvate to pyruvate, is a central metabolic regulator in most organisms. Consequently PK represents an attractive therapeutic target in cancer and human pathogens, like Apicomplexans. The phylum Aplicomplexa, a group of exclusively parasitic organisms, includes the genera Plasmodium, Cryptosporidium and Toxoplasma, the etiological agents of malaria, cryptosporidiosis and toxoplasmosis respectively. Toxoplasma gondii infection causes a mild illness and is a very common infection affecting nearly one third of the world's population. We have determined the crystal structure of the PK1 enzyme from T. gondii, with the B domain in the open and closed conformations. We have also characterized its enzymatic activity and confirmed glucose-6-phosphate as its allosteric activator. This is the first description of a PK enzyme in a closed inactive conformation without any bound substrate. Comparison of the two tetrameric TgPK1 structures indicates a reorientation of the monomers with a concomitant change in the buried surface among adjacent monomers. The change in the buried surface was associated with significant B domain movements in one of the interacting monomers. We hypothesize that a loop in the interface between the A and B domains plays an important role linking the position of the B domain to the buried surface among monomers through two {alpha}-helices. The proposed model links the catalytic cycle of the enzyme with its domain movements and highlights the contribution of the interface between adjacent subunits. In addition, an unusual ordered conformation was observed in one of the allosteric binding domains and it is related to a specific apicomplexan insertion. The sequence and structural particularity would explain the atypical activation by a mono-phosphorylated sugar. The sum of peculiarities raises this enzyme as an emerging target for drug discovery.

  15. Proteomic characterization of the subpellicular cytoskeleton of Toxoplasma gondii tachyzoites.

    Science.gov (United States)

    Gómez de León, Carmen T; Díaz Martín, Rubén Darío; Mendoza Hernández, Guillermo; González Pozos, Sirenia; Ambrosio, Javier R; Mondragón Flores, Ricardo

    2014-12-05

    Toxoplasma, the causative agent of toxoplasmosis in animals and humans, has a subpellicular cytoskeleton that is involved in motility, cell shape and invasion. Knowledge of components of the cytoskeleton is necessary to understand the invasion mechanisms as well as for the identification of possible therapeutic targets. To date, most cytoskeletal components of Toxoplasma remain unidentified due mainly to the lack of reproducible methods for their isolation. Based on the successful isolation of the cytoskeleton, it was possible to report for the first time, the proteomic characterization of the subpellicular cytoskeleton of Toxoplasma formed by 95 cytoskeletal proteins through proteomic analysis by tandem mass spectrometry of one dimension SDS PAGE. By bioinformatic analysis of the data, proteins were classified as: 18 conventional cytoskeletal proteins; 10 inner membrane complex proteins, including 7 with alveolin repeats; 5 new proteins with alveolin like repeats; 37 proteins associated with other organelles and 25 novel proteins of unknown function. One of the alveolin like proteins not previously described in Toxoplasma named TgArticulin was partially characterized with a specific monoclonal antibody. Presence of TgArticulin was exclusively associated with the cytoskeleton fraction with a cortical distribution. Functions for the several molecules identified are proposed. This manuscript describes, for the first time, the proteome of the subpellicular cytoskeleton of Toxoplasma gondii. The importance of this study is related to the role of the cytoskeleton in the highly invasive capability of a parasite that causes abortion, blindness, and death by encephalitis in immunocompromised patients. Proteomic characterization of the cytoskeleton of T. gondii tachyzoites was possible by the development of a successful procedure for the isolation of the subpellicular cytoskeleton. Knowledge of the composition of the cytoskeleton of Toxoplasma is fundamental for the

  16. Seroepidemiology of Toxoplasma gondii Infection in Women of Reproductive Age: A Cross-Sectional Study in a Northwestern Mexican City

    Science.gov (United States)

    Alvarado-Esquivel, Cosme; Corella-Madueno, Maria Alba Guadalupe; Hernandez-Tinoco, Jesus; Rascon-Careaga, Antonio; Sanchez-Anguiano, Luis Francisco; Martinez-Robinson, Karla Guadalupe; Aldana-Madrid, Maria Lourdes; Quizan-Plata, Trinidad; Canez-Carrasco, Maria Guadalupe; Perez-Martinez, Cinthia Jhovanna

    2018-01-01

    Background Through a cross-sectional survey, we determined the seroprevalence and correlates of Toxoplasma gondii (T. gondii) infection in women of reproductive age in Hermosillo City, Mexico. Methods We studied 445 women of reproductive age in Hermosillo City in the northwestern Mexican state of Sonora. Women were enrolled in the University of Sonora. Sera of women were examined for IgG and IgM antibodies to T. gondii by commercially available enzyme immunoassays. The association of T. gondii seropositivity with the characteristics of the pregnant women was determined by bivariate and multivariate analyses. Results Of the 445 women (mean age: 22.18 ± 5.6 years) studied, 16 (3.6%) had IgG antibodies to T. gondii, and two (12.5%) were also positive for IgM antibodies to T. gondii. Of the 16 anti-T. gondii IgG-positive women, six (37.5%) had IgG levels higher than 150 IU/mL, four (25.0%) between 100 and 150 IU/mL, and six (37.5%) between 9 and 99 IU/mL. Multivariate analysis of socio-demographic and behavioral variables showed that T. gondii seropositivity was associated with older age (odds ratio (OR): 5.30; 95% confidence interval (CI): 1.37 - 20.50; P = 0.01) and boar meat consumption (OR: 6.86; 95% CI: 1.27 - 37.07; P = 0.02). Conclusions Women of reproductive age in Hermosillo City had a low seroprevalence of T. gondii infection. However, this finding indicates that most of these women were susceptible to a primary infection. Factors associated with T. gondii infection found in this study may be useful for the optimal planning of preventive measures against T. gondii infection and its sequelae. PMID:29416579

  17. Long-term humoral antibody responses by various serologic tests in pigs orally inoculated with oocysts of four strains of Toxoplasma gondii

    DEFF Research Database (Denmark)

    Dubey, J.P.; Andrews, C.D.; Thulliez, P.

    1997-01-01

    Antibody titers to Toxoplasma gondii were determined in 16 pigs orally inoculated with 1000 or 10000 oocysts of one of the four strains (GT-1, ME-49, TS-2, TC-2) of T. gondii. Pigs were euthanized on postinoculation days 103-875 and their tissues were bioassayed for T. gondii. Antibody titers wer...

  18. Seroprevalence of Anti-Toxoplasma gondii Antibodies Among Lebanese Pregnant Women.

    Science.gov (United States)

    Nahouli, Hasan; El Arnaout, Nour; Chalhoub, Elias; Anastadiadis, Elie; El Hajj, Hiba

    2017-12-01

    Toxoplasma gondii, the causative agent of toxoplasmosis, is a zoonotic obligate intracellular protozoan parasite responsible for the infection of almost one-third of the world's population. T. gondii is particularly threatening for primo-infected pregnant women and may lead, following vertical transplacental transmission, to spontaneous abortion, miscarriage, or severe manifestations in the newborn. The aim of this study was to provide an updated estimate of the seroprevalence of anti-T. gondii antibodies among a group of Lebanese pregnant women and its seroconversion rate. This is a retrospective cohort study, in which medical records of 11,000 pregnant women were screened. These women visited a private Obstetrics and Gynecology clinic located in Beirut, the capital of Lebanon, during the period of January 1994 till September 2015. Serological results of anti-T. gondii immunoglobulin G (IgG) and immunoglobulin M (IgM) results of 2456 Lebanese pregnant women who fulfilled the inclusion criteria were included in the analysis. Seropositivity and seroconversion rates for women with repeated tests were reported according to age and area of residence. The overall anti-T. gondii IgG and IgM seropositivity among 2456 Lebanese pregnant women was 82.6% and 1.8% respectively. The highest IgG seropositivity is among the age group of 35-44 years (87.81%) and at the governorate of "Mount Lebanon" (82.95%). Sixty-four seroconversions were detected and two abortions due to T. gondii infection during pregnancy were recorded. The seroprevalence of anti-T. gondii IgG among the screened pregnant women in Lebanon is the highest in the Arab region. These results highlight the importance of running a national sample survey to estimate the real potential burden of this infection and its impact on maternal and fetal health.

  19. New, previously unreported correlations between latent Toxoplasma gondii infection and excessive ethanol consumption.

    Science.gov (United States)

    Samojłowicz, Dorota; Borowska-Solonynko, Aleksandra; Kruczyk, Marcin

    2017-11-01

    A number of world literature reports indicate that a latent Toxoplasma gondii infection leads to development of central nervous system disorders, which in turn may lead to altered behavior in the affected individuals. T. gondii infection has been observed to play the greatest role in drivers, suicides, and psychiatric patients. Studies conducted for this manuscript involve a different, never before really reported correlation between latent T. gondii infection and ethanol abuse. A total of 538 decedents with a known cause of death were included in the study. These individuals were divided into three groups: the risky behavior group, inconclusively risky behavior group, and control group. The criterion for this division was the likely effect of the individual's behavior on the mechanism and cause of his/her death. The material used for analyses were blood samples collected during routine medico-legal examinations in these cases. The blood samples were used to measure anti-T. gondii IgG antibodies with an enzyme-linked immunosorbent assay (ELISA). Moreover, the following data were recorded for each decedent: sex, age, circumstances of death, cause of death, time from death to autopsy, and (if provided) substance abuse status (alcohol, illicit drugs). In those cases where blood alcohol level or toxicology tests were requested by the Prosecutor's Office, their results were also included in our analysis. Test results demonstrated a strong correlation between latent T. gondii infection and engaging in risky behaviors leading to death. Moreover, analyses demonstrated a positive correlation between the presence of anti-T. gondii IgG antibodies and psychoactive substance (especially ethanol) abuse, however, the causal relationship remains unclear. Due to the fact that alcohol abuse constitutes a significant social problem, searching for eliminable risk factors for addiction is extremely important. Our analyses provided new important information on the possible effects of

  20. Lack of association between Toxoplasma gondii infection and occupational exposure to animals

    Science.gov (United States)

    Pacheco-Vega, Sandy Janet; Hernández-Tinoco, Jesús; Saldaña-Simental, Diana Elizabeth; Sánchez-Anguiano, Luis Francisco; Salcedo-Jáquez, Misael; Ramos-Nevárez, Agar; Liesenfeld, Oliver; Márquez-Conde, José Ángel; Cerrillo-Soto, Sandra Margarita; Martínez-Ramírez, Lucio; Guido-Arreola, Carlos Alberto

    2014-01-01

    The association of infection with Toxoplasma gondii and occupational exposure to animals has been scantly determined. We performed a case-control study with 200 subjects from Durango Province, Mexico, occupationally exposed to animals and 200 age- and gender-matched subjects without this occupation. Sera from all participants were analyzed for anti-T. gondii IgG and IgM antibodies using enzyme-linked immunoassays. The association of seroprevalence with sociodemographic, work, clinical, and behavioral characteristics in cases was determined. Cases and controls had similar frequencies of anti-T. gondii IgG antibodies (12/200: 6.0% and 11/200: 5.5%, respectively) (OR = 3.0; 95% CI: 0.12–73.64; P = 1.0). The frequency of sera with high (>150 IU/ml) levels of anti-T. gondii IgG antibodies was comparable among cases and controls (P = 0.61). Seroprevalence of anti-T. gondii IgM antibodies was similar in cases (4, 2.0%) than in controls (4, 2.0%) (P = 1.0). Multivariate analysis showed that seropositivity was associated with eating while working (OR = 7.14; 95% CI: 1.91–26.72; P = 0.003) and consumption of duck meat (OR = 5.43; 95% CI: 1.43–20.54; P = 0.01). No association between seropositivity to T. gondii and occupational exposure to animals was found. However, risk factors for infection found should be taken into account to reduce the exposure to T. gondii. PMID:25544890

  1. Mouse-virulent Toxoplasma gondii isolated from feral cats on Mona Island, Puerto Rico.

    Science.gov (United States)

    Dubey, J P; López-Torres, H Y; Sundar, N; Velmurugan, G V; Ajzenberg, D; Kwok, O C H; Hill, R; Dardé, M L; Su, C

    2007-12-01

    Cats are essential in the life cycle of Toxoplasma gondii because they are the only hosts that can excrete the environmentally resistant oocysts. Samples of serum, feces, and tissues from cats from Mona, a remote island off the coast of Puerto Rico, were examined for T. gondii infection. Antibodies to T. gondii were assayed by the modified agglutination test and found in 16 of 19 (84.2%) of cats, with titers of 1:10 in 2, 1:80 in 1, 1:160 in 4, 1:320 in 3, and 1:1,280 or higher in 6. Tissues of 19 of the 20 cats were bioassayed in mice for T. gondii infection. Toxoplasma gondii was isolated from tissues of 12 cats: from the hearts of 9, skeletal muscle of 10, and brain of 1 cat. All infected mice from 10 of 12 isolates died of acute toxoplasmosis during primary infection. Genotyping of these 12 T. gondii isolates (designated (TgCatPr 1-12) by 10 multilocus PCR-RFLP markers, i.e., SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and an apicoplast marker Apico, and the 6 multilocus microsatellite markers TUB2, W35, TgM-A, B18, B17, and M33, revealed 7 genotypes; 5 isolates had Type I alleles at all loci except at 1 microsatellite locus, and the remainder were atypical. The latter isolates of T. gondii were different biologically and phenotypically from the feline isolates from the rest of the Americas. One isolate (TgCatPr 12) was a mixed infection with 2 genotypes.

  2. An experimental Toxoplasma gondii dose response challenge model to study therapeutic or vaccine efficacy in cats.

    Directory of Open Access Journals (Sweden)

    Jan B W J Cornelissen

    Full Text Available High numbers of Toxoplasma gondii oocysts in the environment are a risk factor to humans. The environmental contamination might be reduced by vaccinating the definitive host, cats. An experimental challenge model is necessary to quantitatively assess the efficacy of a vaccine or drug treatment. Previous studies have indicated that bradyzoites are highly infectious for cats. To infect cats, tissue cysts were isolated from the brains of mice infected with oocysts of T. gondii M4 strain, and bradyzoites were released by pepsin digestion. Free bradyzoites were counted and graded doses (1000, 100, 50, 10, and 250 intact tissue cysts were inoculated orally into three cats each. Oocysts shed by these five groups of cats were collected from faeces by flotation techniques, counted microscopically and estimated by real time PCR. Additionally, the number of T. gondii in heart, tongue and brains were estimated, and serology for anti T. gondii antibodies was performed. A Beta-Poisson dose-response model was used to estimate the infectivity of single bradyzoites and linear regression was used to determine the relation between inoculated dose and numbers of oocyst shed. We found that real time PCR was more sensitive than microscopic detection of oocysts, and oocysts were detected by PCR in faeces of cats fed 10 bradyzoites but by microscopic examination. Real time PCR may only detect fragments of T. gondii DNA without the presence of oocysts in low doses. Prevalence of tissue cysts of T. gondii in tongue, heart and brains, and anti T. gondii antibody concentrations were all found to depend on the inoculated bradyzoite dose. The combination of the experimental challenge model and the dose response analysis provides a suitable reference for quantifying the potential reduction in human health risk due to a treatment of domestic cats by vaccination or by therapeutic drug application.

  3. Aborto espontâneo e toxoplasmose ocular em um casal infectado com Toxoplasma gondii = Spontaneous abortion and ocular toxoplasmosis in a couple infected with Toxoplasma gondii

    Directory of Open Access Journals (Sweden)

    Souza, Carla Zangari de

    2015-01-01

    Full Text Available Objetivos: Relatar os casos de um casal, em que a esposa apresentou toxoplasmose na gestação e o esposo, após 15 anos, foi diagnosticado com toxoplasmose ocular. Relato dos casos: Gestante, grávida de gêmeos, residente de um estado na região Sul do Brasil. No primeiro mês de gestação apresentou sorologia não reagente para IgG e IgM anti-Toxoplasma gondii. Na sétima semana de gestação houve soroconversão (IgM e IgG anti-T. gondii reagentes e aborto natural. O esposo, após 15 anos desde o episódio da esposa, queixou-se de visão embaçada/irritada. O exame de fundoscopia revelou lesões típicas de coriorretinite recidivante por toxoplasmose no olho direito. Os exames sorológicos foram IgG anti-T. gondii reagente e IgM anti-T. gondii não reagente. Conclusões: Este relato, de um casal com desfechos clínicos importantes de toxoplasmose, em diferentes formas, enfatiza a relevância do problema da toxoplasmose, que nas suas diversas manifestações, pode trazer graves consequências para a vida de uma família

  4. Toxoplasma gondii seroprevalence in donkeys (Equus asinus) from the U.S.A. and isolation of T. gondii from cats on donkey farms

    Science.gov (United States)

    Donkeys (Equus asinus) are used as companion animals, in leisure activities as working animals, for onotherapy, and their meat and milk is used for human consumption in certain countries. Here we report first serological survey for Toxoplasma gondii in donkeys in the U.S.A. Serum samples were collec...

  5. Characterisation of Toxoplasma gondii isolates using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) of the non-coding Toxoplasma gondii (TGR)-gene sequences

    DEFF Research Database (Denmark)

    Høgdall, Estrid; Vuust, Jens; Lind, Peter

    2000-01-01

    of using TGR gene variants as markers to distinguish among T. gondii isolates from different animals and different geographical sources. Based on the band patterns obtained by restriction fragment length polymorphism (RFLP) analysis of the polymerase chain reaction (PCR) amplified TGR sequences, the T...

  6. Cyclosporin A treatment of Leishmania donovani reveals stage-specific functions of cyclophilins in parasite proliferation and viability.

    Directory of Open Access Journals (Sweden)

    Wai-Lok Yau

    Full Text Available BACKGROUND: Cyclosporin A (CsA has important anti-microbial activity against parasites of the genus Leishmania, suggesting CsA-binding cyclophilins (CyPs as potential drug targets. However, no information is available on the genetic diversity of this important protein family, and the mechanisms underlying the cytotoxic effects of CsA on intracellular amastigotes are only poorly understood. Here, we performed a first genome-wide analysis of Leishmania CyPs and investigated the effects of CsA on host-free L. donovani amastigotes in order to elucidate the relevance of these parasite proteins for drug development. METHODOLOGY/PRINCIPAL FINDINGS: Multiple sequence alignment and cluster analysis identified 17 Leishmania CyPs with significant sequence differences to human CyPs, but with highly conserved functional residues implicated in PPIase function and CsA binding. CsA treatment of promastigotes resulted in a dose-dependent inhibition of cell growth with an IC50 between 15 and 20 microM as demonstrated by proliferation assay and cell cycle analysis. Scanning electron microscopy revealed striking morphological changes in CsA treated promastigotes reminiscent to developing amastigotes, suggesting a role for parasite CyPs in Leishmania differentiation. In contrast to promastigotes, CsA was highly toxic to amastigotes with an IC50 between 5 and 10 microM, revealing for the first time a direct lethal effect of CsA on the pathogenic mammalian stage linked to parasite thermotolerance, independent from host CyPs. Structural modeling, enrichment of CsA-binding proteins from parasite extracts by FPLC, and PPIase activity assays revealed direct interaction of the inhibitor with LmaCyP40, a bifunctional cyclophilin with potential co-chaperone function. CONCLUSIONS/SIGNIFICANCE: The evolutionary expansion of the Leishmania CyP protein family and the toxicity of CsA on host-free amastigotes suggest important roles of PPIases in parasite biology and implicate

  7. Latitudinal variability in the seroprevalence of antibodies against Toxoplasma gondii in non-migrant and Arctic migratory geese

    NARCIS (Netherlands)

    Sandstrom, Cecilia A. M.; Buma, Anita G. J.; Hoye, Bethany J.; Prop, Jouke; van der Jeugd, Henk; Voslamber, Berend; Madsen, Jesper; Loonen, Maarten J. J. E.

    2013-01-01

    Toxoplasma gondii is an intracellular coccidian parasite found worldwide and is known to infect virtually all warm-blooded animals. It requires a cat (family Felidae) to complete its full life cycle. Despite the absence of wild felids on the Arctic archipelago of Svalbard, T. gondii has been found

  8. Comparative genomic sequence variation of Toxoplasma gondii reveals local admixture drives concerted expansion and diversification of secreted pathogenesis determinants

    Science.gov (United States)

    Toxoplasma gondii is among the most abundant parasites world-wide, infecting many wild and domestic animals and causing zoonotic infections in humans. T. gondii differs substantially in its distribution from closely related parasites that typically have narrow, specialized host ranges. We undertook...

  9. Comparative sequence analysis of Toxoplasma gondii reveals local genomic admixture drives concerted expansion and diversification of secreted pathogenesis determinants

    Science.gov (United States)

    Toxoplasma gondii is among the most prevalent parasites worldwide, infecting many wild and domestic animals and causing zoonotic infections in humans. T. gondii differs substantially in its broad distribution from closely related parasites that typically have narrow, specialized host ranges. To un...

  10. Toxoplasmosis in geese and detection of two new atypical Toxoplasma gondii strains from naturally infected Canada geese (Branta canadensis)

    Science.gov (United States)

    The protozoan Toxoplasma gondii infects virtually all warm-blooded animals, including birds, humans, livestock, and marine mammals. The consumption of raw or undercooked meat infected with T. gondii is considered an important source of infection in humans. Canada goose (Branta canadensis), the most ...

  11. Detection of Toxoplasma gondii DNA in horse meat from supermarkets in France and performance evaluation of two serological tests

    Directory of Open Access Journals (Sweden)

    Aroussi Abdelkrim

    2015-01-01

    Full Text Available In France, some cases of severe toxoplasmosis have been linked to the consumption of horse meat that had been imported from the American continent where atypical strains of Toxoplasma gondii are more common than in Europe. Many seroprevalence studies are presented in the literature but risk assessment of T. gondii infection after horse meat consumption is not possible in the absence of validated serological tests and the unknown correlation between detection of antibodies against T. gondii and presence of tissue cysts. We performed magnetic-capture polymerase chain reaction (MC-PCR to detect T. gondii DNA in 231 horse meat samples purchased in supermarkets in France and evaluated the performance and level of agreement of the modified agglutination test (MAT and enzyme-linked immunosorbent assay (ELISA in the meat juices. The serological tests lacked sensitivity, specificity, and agreement between them, and there was no correlation with the presence of T. gondii DNA in horse meat, raising concerns about the reliability of T. gondii seroprevalence data in horses from the literature. T. gondii DNA was detected in 43% of horse meat samples but the absence of strain isolation in mice following inoculation of more than 100 horse meat samples suggests a low distribution of cysts in skeletal muscles and a low risk of T. gondii infection associated with horse meat consumption. However, to avoid any risk of toxoplasmosis, thorough cooking of horse meat is recommended.

  12. Detection of Toxoplasma gondii DNA in horse meat from supermarkets in France and performance evaluation of two serological tests.

    Science.gov (United States)

    Aroussi, Abdelkrim; Vignoles, Philippe; Dalmay, François; Wimel, Laurence; Dardé, Marie-Laure; Mercier, Aurélien; Ajzenberg, Daniel

    2015-01-01

    In France, some cases of severe toxoplasmosis have been linked to the consumption of horse meat that had been imported from the American continent where atypical strains of Toxoplasma gondii are more common than in Europe. Many seroprevalence studies are presented in the literature but risk assessment of T. gondii infection after horse meat consumption is not possible in the absence of validated serological tests and the unknown correlation between detection of antibodies against T. gondii and presence of tissue cysts. We performed magnetic-capture polymerase chain reaction (MC-PCR) to detect T. gondii DNA in 231 horse meat samples purchased in supermarkets in France and evaluated the performance and level of agreement of the modified agglutination test (MAT) and enzyme-linked immunosorbent assay (ELISA) in the meat juices. The serological tests lacked sensitivity, specificity, and agreement between them, and there was no correlation with the presence of T. gondii DNA in horse meat, raising concerns about the reliability of T. gondii seroprevalence data in horses from the literature. T. gondii DNA was detected in 43% of horse meat samples but the absence of strain isolation in mice following inoculation of more than 100 horse meat samples suggests a low distribution of cysts in skeletal muscles and a low risk of T. gondii infection associated with horse meat consumption. However, to avoid any risk of toxoplasmosis, thorough cooking of horse meat is recommended. © A. Aroussi et al., published by EDP Sciences, 2015.

  13. New Toxoplasma gondii genotypes isolated from free-range chickens from the Fernando de Noronha, Brazil: unexpected findings

    Science.gov (United States)

    Worldwide comparison of Toxoplasma gondii isolates from free range chickens has indicated that T. gondii isolates from Brazil are phenotypically and genetically different than isolates from other countries; most strains from Brazil are pathogenic to mice, there is great genetic variability, most iso...

  14. Diagnosis of pulmonary infection with Toxoplasma gondii in immunocompromised HIV-positive patients by real-time PCR

    DEFF Research Database (Denmark)

    Petersen, E; Edvinsson, B; Lundgren, B

    2006-01-01

    The aim of the study presented here was to evaluate the use of PCR for improving the diagnosis of Toxoplasma gondii infection in immunocompromised hosts. Three hundred thirty-two bronchoalveolar lavage (BAL) fluid samples were analyzed by real-time PCR targeting a 529 bp element of T. gondii. In ...

  15. Genetic characterization of Toxoplasma gondii isolates from Portugal, Austria, and Israel reveals higher genetic variability within the type II lineage

    Science.gov (United States)

    This study compared genetic diversity of Toxoplasma gondii isolates from Portugal, Austria and Israel. For this, we genotyped 90 T. gondii isolates (16 from Portugal, 67 from Austria and 7 from Israel) using 10 nested PCR-restriction length polymorphism (RFLP) genetic markers and 15 microsatellite (...

  16. Occurrence of Toxoplasma gondii antibodies in wild rodents and marsupials from the Atlantic forest, State of Sao Paulo, Brazil

    Science.gov (United States)

    Toxoplasma gondii is a protozoan parasite that infects a large spectrum of warm-blooded animals, including humans. Small mammals and rodents play an important role in the epidemiology of T. gondii because they are sources of infection for domestic and feral cats. Serum samples from 151 rodents and 4...

  17. Seroprevalence of Toxoplasma gondii in wild marsupials and rodents from the Atlantic forest of Pernambuco State, Northeastern region, Brazil

    Science.gov (United States)

    Felids are important in the epidemiology of Toxoplasma gondii because they are the only hosts that can excrete the environmentally resistant oocysts in their feces. Cats are considered to acquire T. gondii infection in nature by ingesting tissues of small mammals and birds. Serum samples of feral 22...

  18. Production, characterization and applications for Toxoplasma gondii-specific polyclonal chicken egg yolk immunoglobulins.

    Directory of Open Access Journals (Sweden)

    Álvaro Ferreira Júnior

    Full Text Available Toxoplasma gondii may cause abortions, ocular and neurological disorders in warm-blood hosts. Immunized mammals are a wide source of hyperimmune sera used in different approaches, including diagnosis and the study of host-parasite interactions. Unfortunately, mammalian antibodies present limitations for its production, such as the necessity for animal bleeding, low yield, interference with rheumatoid factor, complement activation and affinity to Fc mammalian receptors. IgY antibodies avoid those limitations; therefore they could be an alternative to be applied in T. gondii model.In this study we immunized hens with soluble tachyzoite antigens of T. gondii (STAg and purified egg yolk antibodies (IgY by an inexpensive and simple method, with high yield and purity degree. IgY anti-STAg antibodies presented high avidity and were able to recognize a broad range of parasite antigens, although some marked differences were observed in reactivity profile between antibodies produced in immunized hens and mice. Interestingly, IgY antibodies against Neospora caninum and Eimeria spp. did not react to STAg. We also show that IgY antibodies were suitable to detect T. gondii forms in paraffin-embedded sections and culture cell monolayers.Due to its cost-effectiveness, high production yield and varied range of possible applications, polyclonal IgY antibodies are useful tools for studies involving T. gondii.

  19. Toxoplasma gondii exposes phosphatidylserine inducing a TGF-β1 autocrine effect orchestrating macrophage evasion

    International Nuclear Information System (INIS)

    Seabra, Sergio H.; Souza, Wanderley de; Matta, Renato A. da

    2004-01-01

    Toxoplasmosis is a worldwide disease caused by Toxoplasma gondii. Activated macrophages control T. gondii growth by nitric oxide (NO) production. However, T. gondii active invasion inhibits NO production, allowing parasite persistence. Here we show that the mechanism used by T. gondii to inhibit NO production persisting in activated macrophages depends on phosphatidylserine (PS) exposure. Masking PS with annexin-V on parasites or activated macrophages abolished NO production inhibition and parasite persistence. NO production inhibition depended on a transforming growth factor-β 1 (TGF-β 1 ) autocrine effect confirmed by the expression of Smad 2 and 3 in infected macrophages. TGF-β 1 led to inducible nitric oxide synthase (iNOS) degradation, actin filament (F-actin) depolymerization, and lack of nuclear factor-κB (NF-κB) in the nucleus. All these features were reverted by TGF-β 1 neutralizing antibody treatment. Thus, T. gondii mimics the evasion mechanism used by Leishmania amazonensis and also the anti-inflammatory response evoked by apoptotic cells

  20. Atypical Toxoplasma gondii genotype in feral cats from the Fernando de Noronha Island, northeastern Brazil.

    Science.gov (United States)

    Melo, R P B; Almeida, J C; Lima, D C V; Pedrosa, C M; Magalhães, F J R; Alcântara, A M; Barros, L D; Vieira, R F C; Garcia, J L; Mota, R A

    2016-07-15

    Toxoplasma gondii isolates from Brazil have a different phenotypic and genotypic pattern, with predominance of virulent isolates and recombinant genotypes, compared to the North Hemisphere. Considering that a new T. gondii genotype, non-pathogenic to mice, was previously identified from free-range chickens from the Fernando de Noronha Island, Brazil, this study aimed to identify genotypes of this parasite in tissue samples of feral cats (Felis catus) from this Brazilian Island. Anti-T. gondii IgG antibodies were detected in 18/31 (58%) feral cats. Two non-virulent T. gondii isolates were obtained by mouse bioassay. Genotyping was performed by PCR-RFLP using 10 genetic markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, PK1, L358 and Apico) and an atypical strain of T. gondii (ToxoDB #146) was identified. This is the first report of this genotype in feral cats. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Seroprevalence of Toxoplasma gondii in wild boars, red deer and roe deer in Poland

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    Witkowski Lucjan

    2015-01-01

    Full Text Available Little is known about the prevalence of Toxoplasma gondii in wild life, particularly game animals in Poland. Meat juice collected during the 2009/2010 and 2010/2011 hunting seasons from 552 red deer (Cervus elaphus, 367 wild boars (Sus scrofa and 92 roe deer (Capreolus capreolus was tested for T. gondii antibodies using the multi-species ID Screen Toxoplasmosis Indirect kit (IDvet, Montpellier, France. Antibodies to T. gondii were detected in 24.1% of red deer (95% CI: 20.7%, 27.8%, 37.6% of wild boar (95% CI: 32.8%, 42.7% and 30.4% of roe deer (95% CI: 22.0%, 40.5%. To the authors’ best knowledge, this is the first epidemiological report of T. gondii prevalence in red deer, roe deer and wild boars in Poland. T. gondii is present in wildlife animal tissues and consumption of the game may be a potential source of infection for humans.

  2. Fatal toxoplasmosis in an immunosuppressed domestic cat from Brazil caused by Toxoplasma gondii clonal type I

    Directory of Open Access Journals (Sweden)

    Hilda Fátima de Jesus Pena

    Full Text Available Abstract The objective of the study was to report on a fatal case of feline toxoplasmosis with coinfection with the feline leukemia virus (FeLV. A domestic cat (Felis silvestris catus presented intense dyspnea and died three days later. In the necropsy, the lungs were firm, without collapse and with many white areas; moderate lymphadenomegaly and splenomegaly were also observed. The histopathological examination showed severe necrotic interstitial bronchopneumonia and mild necrotic hepatitis, associated with intralesional cysts and tachyzoites of Toxoplasma gondii that were positive by anti-T. gondii immunohistochemical (IHC evaluation. The bone marrow showed chronic myeloid leukemia and the neoplastic cells were positive by anti-FeLV IHC evaluation. DNA extracted from lungs was positive for T. gondii by PCR targeting REP-529. T. gondii was characterized by PCR-RFLP and by the microsatellites technique. ToxoDB-PCR-RFLP #10, i.e. the archetypal type I, was identified. Microsatellite analysis showed that the strain was a variant of type I with two atypical alleles. This was the first time that a T. gondii clonal type I genotype was correlated with a case of acute toxoplasmosis in a host in Brazil.

  3. Low Seroprevalence of Leishmania infantum and Toxoplasma gondii in the Horse Population in Israel.

    Science.gov (United States)

    Aharonson-Raz, Karin; Baneth, Gad; Lopes, Ana Patrícia; Brancal, Hugo; Schallig, Henk; Cardoso, Luís; Steinman, Amir

    2015-12-01

    A cross-sectional investigation was done on the seroprevalence of Leishmania infantum and Toxoplasma gondii infection among apparently healthy horses in Israel. This survey included 383 horses distributed in 22 farms throughout Israel during the years 2011-2013. Serum samples were tested for the presence of immunoglobulin G (IgG) antibodies using the direct agglutination test (DAT) specific to Leishmania and by the modified agglutination test (MAT) for the presence of IgG antibodies to T. gondii. Low seroprevalences were detected for both L. infantum and T. gondii in the horse population in Israel; of the 338 horses tested, 6 (1.4%) were found to be seropositive for L. infantum and 11 (2.5%) for T. gondii, with no significant association between seroprevalence and demographic/environmental factors. An ongoing geographical expansion of L. infantum, previously reported in humans and dogs in Israel, was also supported by our results in horses. Here we present evidence of exposure of horses to L. infantum and T. gondii in Israel. Continuous seroprevalence surveillance in horses, such as the one performed in this study, might further elucidate the eco-epidemiology of these two important zoonotic parasites in this country.

  4. Occurrence of Toxoplasma gondii antibodies in lowland tapirs maintained ex situ in Brazil and Paraguay

    Directory of Open Access Journals (Sweden)

    Maria Fernanda Naegeli Gondim

    Full Text Available ABSTRACT: Lowland Tapir ( Tapirus terrestris is the second largest South American land mammal. It is strictly herbivorous and its exposure to Toxoplasma gondii should be indicative of environmental contamination by oocysts.In the present study antibodies to T. gondii in 47 Brazilian tapirs maintained ex situ in 10 Brazilian and in one Paraguayan institution were sought in serum samples by the modified agglutination test (MAT ≥25. None of the animals presented clinical signs during the study. From 47 animals 35 (74.5% were positive with titers of 25 in 8, 50 in 6, 100 in 12, 200 in 5, 400 in 1 and 800 in 3. One animal had samples collected on twice, and 19 were born in captivity. There was no association between occurrence of T. gondii antibodies and gender, and positive animals were reported in all institutions. The high occurrence of seropositive tapirs born ininstitutions (54.3% confirmed the high exposure of these mammals to T. gondii in captivity. Only two cases ofabortion were reported, but it was not possible to correlate these abortions to T. gondii infection.

  5. Host Mitochondrial Association Evolved in the Human Parasite Toxoplasma gondii via Neofunctionalization of a Gene Duplicate.

    Science.gov (United States)

    Adomako-Ankomah, Yaw; English, Elizabeth D; Danielson, Jeffrey J; Pernas, Lena F; Parker, Michelle L; Boulanger, Martin J; Dubey, Jitender P; Boyle, Jon P

    2016-05-01

    In Toxoplasma gondii, an intracellular parasite of humans and other animals, host mitochondrial association (HMA) is driven by a gene family that encodes multiple mitochondrial association factor 1 (MAF1) proteins. However, the importance of MAF1 gene duplication in the evolution of HMA is not understood, nor is the impact of HMA on parasite biology. Here we used within- and between-species comparative analysis to determine that the MAF1 locus is duplicated in T. gondii and its nearest extant relative Hammondia hammondi, but not another close relative, Neospora caninum Using cross-species complementation, we determined that the MAF1 locus harbors multiple distinct paralogs that differ in their ability to mediate HMA, and that only T. gondii and H. hammondi harbor HMA(+) paralogs. Additionally, we found that exogenous expression of an HMA(+) paralog in T. gondii strains that do not normally exhibit HMA provides a competitive advantage over their wild-type counterparts during a mouse infection. These data indicate that HMA likely evolved by neofunctionalization of a duplicate MAF1 copy in the common ancestor of T. gondii and H. hammondi, and that the neofunctionalized gene duplicate is selectively advantageous. Copyright © 2016 by the Genetics Society of America.

  6. Seroprevalence of Toxoplasma gondii and Neospora caninum infections in goats from two Argentinean provinces

    Directory of Open Access Journals (Sweden)

    María Laura Gos

    2017-11-01

    Full Text Available The aim of this study was to compare the seroprevalence of Toxoplasma gondii and Neospora caninum in goats from two Argentinean provinces raised under different management conditions. A total of 2922 serum samples from adult goats of Córdoba (n=2187 and Buenos Aires provinces (n= 735, Argentina, were assayed by indirect fluorescence antibody test (IFAT, cut-off 1:100 for antibodies to T. gondii and N. caninum. Seroprevalence was 40.8% (CI 39.0%-42.6% and 5.5% (CI 4.7%-6.4% for T. gondii and N. caninum, respectively. The seropositivity for both infections was higher in goats from dairy farms, resulting as follows: for T. gondii 32.7% (CI 30.8%-34.8% in extensive farms and 59.3% (CI 56.1%-62.6% in dairy farms and for N. caninum 4.1% (CI 3.2%-4.9% in extensive farms and 8.8% (CI 6.9%-10.7% in dairy farms. This is the first extensive seroepidemiology investigation for T. gondii and N. caninum in goats in Argentina.

  7. Portugal and Angola: similarities and differences in Toxoplasma gondii seroprevalence and risk factors in pregnant women.

    Science.gov (United States)

    Lobo, M L; Patrocinio, G; Sevivas, T; DE Sousa, B; Matos, O

    2017-01-01

    In this study we determined the presence of IgM/IgG antibodies to Toxoplasma gondii in sera of 155 and 300 pregnant women from Lisbon (Portugal) and Luanda (Angola), respectively, and evaluated the potential risk factors associated with this infection. DNA detection was performed by PCR assays targeting T. gondii regions (RE/B1). Overall, 21·9% (10·9% IgG, 10·9% IgG/IgM) of the Lisbon women and 27·3% (23·7%, IgG, 2% IgM, 1·7% IgG/IgM) of the Luanda women had antibodies to T. gondii. Single variable and binary logistic regression analyses were conducted. Based on the latter, contacts with cats (family/friends), and having more than two births were identified as risk factors for Toxoplasma infection in Lisbon women. In Luanda, the risk factors for T. gondii infection suggested by the single variable analysis (outdoor contact with cats and consumption of pasteurized milk/dairy products) were not confirmed by binary logistic regression. This study shows original data from Angola, and updated data from Portugal in the study of infection by T. gondii in pregnant women, indicating that the prevalence of anti-Toxoplasma antibodies is high enough to alert the government health authorities and implement appropriate measures to control this infection.

  8. Occurrence of antibodies to Toxoplasma gondii in rheas (Rhea americana and ostriches (Struthio camelus from farms of different Brazilian regions

    Directory of Open Access Journals (Sweden)

    Amanda Brentano Almeida

    Full Text Available This study aimed to verify the occurrence of antibodies againstToxoplasma gondii in rheas (Rhea americana and ostriches (Struthio camelus commercially breeding in Brazil. Blood samples from 20 rheas and 46 ostriches (young and adults were serologically tested using a technique known as modified agglutination test (MAT at an initial titration of 1:16 for ostriches and 1:25 for rheas. Antibodies against T. gondii were found in 50% (10/20 of the rheas, with titers ranging from 1:25 to 1:6,400. The incidence of antibodies against T. gondii in ostriches was 17.4% (8/46 with titers ranging from 1:16 to 1:256. Birds showing titers higher than 1:200 forT. gondii were mainly the young ones. Therefore, rheas and ostriches may be parasitized by T. gondii, showing high levels of antibodies against this parasite.

  9. Seroprevalence and Risk Factors of Toxoplasma gondii Infection in Buffaloes, Sheep and Goats in Yunnan Province, Southwestern China

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    Fengcai ZOU

    2015-12-01

    Full Text Available Background: The seroprevalence of Toxoplasma gondii infection in buffaloes, sheep and goats in Yunnan Province, southwestern China was conducted between May 2012 and December 2013.Methods: A total of 973 (427 buffaloes, 154 sheep and 392 goats serum samples were collected from seven administrative regions of Yunnan Province, and exam­ined for T. gondii antibodies by indirect hemagglutination (IHA test. Some risk fac­tors related to species, age, gender and geographical origin were determined using a multinomial logistic regression.Results: The overall seroprevalence of T. gondii in ruminant species was estimated at 11.9%. The final logistic regression model demonstrated that host species and geographical origin were the main risk factors associated with T. gondii infection (P﹤0.05.Conclusion: Taken together, the results of the present study revealed a high expo­sure to T. gondii in ruminant species in Yunnan Province, which has an important implication for public health.

  10. Prevalence of Toxoplasma gondii infection in HIV-infected patients and food animals and direct genotyping of T. gondii isolates, Southern Ghana.

    Science.gov (United States)

    Pappoe, Faustina; Cheng, Weisheng; Wang, Lin; Li, Yuanling; Obiri-Yeboah, Dorcas; Nuvor, Samuel Victor; Ambachew, Henock; Hu, Xiaodong; Luo, Qingli; Chu, Deyong; Xu, Yuanhong; Shen, Jilong

    2017-06-01

    Toxoplasma gondii is of public health and veterinary importance causing severe diseases in immunocompromised individuals including HIV/AIDS patients and in congenital cases and animals. There is limited information on the epidemiology of T. gondii infection in humans, particularly HIV patients and food animals and the parasite genotypes in Ghana. A total of 394 HIV-infected patients from three hospitals were screened for T. gondii anti-IgG and IgM using ELISA. DNAs from blood samples of seropositve participants and 95 brain tissues of food animals were PCR assayed to detect Toxoplasma gra6. DNA positive samples were genotyped using multilocus nested polymerase chain reaction restriction fragment length polymorphism at 10 loci: sag1, alt.sag2, sag3, btub, gra6, l358, c22-8, c29-2, pk1, and apico. The overall seroprevalence was 74.37% (293/394). Toxoplasma DNAs were detected in 3.07% of the seropositive participants and 9.47% of the animals. Six of the human DNA positive samples were partly typed at sag3: 33.33, 50, and 16.67% isolates had type I, II, and III alleles, respectively. All nine isolates from food animals typed at nine loci except apico were atypical: six isolates were identical to ToxoDB #41 and #145, and one was identical to TgCkBrRj2 all identified in Brazil. The genotype of two isolates has not been reported previously and was named as TgCtGh1. T. gondii seroprevalence is high among the HIV-infected individuals with T. gondii circulating in Ghana being genetically diverse.

  11. Dual Identification and Analysis of Differentially Expressed Transcripts of Porcine PK-15 Cells and Toxoplasma gondii during in vitro Infection

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    Chun-Xue eZhou

    2016-05-01

    Full Text Available Toxoplasma gondii is responsible for causing toxoplasmosis, one of the most prevalent zoonotic parasitoses worldwide. The mechanisms that mediate T. gondii infection of pigs (the most common source of human infection and renal tissues are still unknown. To identify the critical alterations that take place in the transcriptome of both porcine kidney (PK-15 cells and T. gondii following infection, infected cell samples were collected at 1, 3, 6, 9, 12, 18, and 24 h post infection and RNA-Seq data were acquired using Illumina Deep Sequencing. Differential Expression of Genes (DEGs analysis was performed to study the concomitant gene-specific temporal patterns of induction of mRNA expression of PK-15 cells and T. gondii. High sequence coverage enabled us to thoroughly characterize T. gondii transcriptome and identify the activated molecular pathways in host cells. More than 6G clean bases/ sample, including > 40 million clean reads were obtained. These were aligned to the reference genome of T. gondii and wild boar (Sus scrofa. DEGs involved in metabolic activities of T. gondii showed time-dependent down-regulation. However, DEGs involved in immune or disease related pathways of PK-15 cells peaked at 6 h PI, and were highly enriched as evidenced by KEGG analysis. Protein-protein interaction analysis revealed that TGME49_120110 (PCNA, TGME49_049180 (DHFR-TS, TGME49_055320 and TGME49_002300 (ITPase are the four hub genes with most interactions with T. gondii at the onset of infection. These results reveal altered profiles of gene expressed by PK-15 cells and T. gondii during infection and provide the groundwork for future virulence studies to uncover the mechanisms of T. gondii interaction with porcine renal tissue by functional analysis of these DEGs.

  12. Reciprocal moderation by Toxoplasma gondii seropositivity and blood phenylalanine - tyrosine ratio of their associations with trait aggression.

    Science.gov (United States)

    Mathai, Ashwin Jacob; Lowry, Christopher A; Cook, Thomas B; Brenner, Lisa A; Brundin, Lena; Groer, Maureen W; Peng, Xiaoqing; Giegling, Ina; Hartmann, Annette M; Konte, Bettina; Friedl, Marion; Fuchs, Dietmar; Rujescu, Dan; Postolache, Teodor T

    2016-12-01

    We previously reported that trait aggression, proposed as an endophenotype for suicidal behavior, is positively associated with Toxoplasma gondii ( T. gondii ) seropositivity in females, but not in males. Additionally, older males seropositive for T. gondii had lower scores on measures of trait aggression, including self-aggression. Trait aggression may be influenced by dopaminergic signaling, which is known to be moderated by gender and age, and potentially enhanced in T. gondii positives through the intrinsic production of dopamine by the microorganism. Therefore, we investigated associations between trait aggression and interactions between T. gondii enzyme-linked immunoabsorbant assay (ELISA) IgG titer-determined seropositivity and high-performance liquid chromatography- (HPLC-) measured blood levels of dopamine precursors phenylalanine (Phe), tyrosine (Tyr), and their ratio in a sample of 1000 psychiatrically healthy participants. Aggressive traits were assessed using the questionnaire for measuring factors of aggression (FAF), the German version of the Buss-Durkee hostility questionnaire. We found that 1) the decrease in trait aggression scores in T. gondii -positive older males was only present in individuals with a low Phe:Tyr ratio, and 2) that there was a positive correlation between Phe:Tyr ratio and total aggression and selected subscales of aggression in T. gondii -positive males, but not in T. gondii -negative males. These findings point toward a gender-specific reciprocal moderation by Phe:Tyr ratio and T. gondii seropositivity of their associations with aggression scores, and lead to experimental interventions geared to manipulating levels of dopamine precursors in selected T. gondii positive individuals with increased propensity for aggression.

  13. Genotyping of samples from German patients with ocular, cerebral and systemic toxoplasmosis reveals a predominance of Toxoplasma gondii type II.

    Science.gov (United States)

    Herrmann, Daland C; Maksimov, Pavlo; Hotop, Andrea; Groß, Uwe; Däubener, Walter; Liesenfeld, Oliver; Pleyer, Uwe; Conraths, Franz J; Schares, Gereon

    2014-10-01

    Toxoplasmosis is an important zoonosis transmitted from animals to humans world-wide. In order to determine Toxoplasma gondii genotypes in individuals living in Germany and to compare findings with those in animals, we analysed nine independent and unlinked genetic markers (nSAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico) by PCR-RFLP in 83 archived T. gondii-positive DNA samples from patients with ocular toxoplasmosis (n=35), toxoplasmic encephalitis (n=32), systemic toxoplasmosis after bone-marrow transplantation (n=15) and congenital toxoplasmosis (n=1). In 46 of these 83 samples the presence of T. gondii DNA was confirmed by conventional end-point PCR. Among these, 17 T. gondii-positive samples were typed at all nine loci. The majority (15/17, 88.2%) of these samples were of T. gondii type II (i.e., including both, the Apico type II and Apico type I variants). In addition, in one sample a T. gondii type II/type III allele combination and in another sample a T. gondii genotype displaying type III alleles at all markers was observed. In the remaining 11 samples, in which T. gondii could only be partially typed, exclusively type II (n=10) or type III (n=1) alleles were observed. Results of the present study suggest that the majority of patients in Germany are infected with type II T. gondii regardless of the clinical manifestation of toxoplasmosis. This finding is in accord with the predominance of type II T. gondii in oocysts isolated from cats and in tissues of other intermediate hosts in Germany. Copyright © 2014 Elsevier GmbH. All rights reserved.

  14. Detection of Toxoplasma gondii oocysts in soils in northwestern China using a new semi-nested PCR assay.

    Science.gov (United States)

    Wang, Meng; Meng, Peng; Ye, Qiang; Pu, Yuan-Hua; Yang, Xiao-Yu; Luo, Jian-Xun; Zhang, Nian-Zhang; Zhang, De-Lin

    2014-09-28

    Toxoplasma gondii is a zoonotic pathogen that can infect a range of animals and humans. Ingestion of T. gondii oocysts in soil is a significant transmission route for humans and animals acquiring toxoplasmosis. In the present study, we developed a new semi-nested PCR method to determine T. gondii oocysts distribution in soils in northwestern China. The one tube semi-nested PCR assay was developed to detect the oocysts of T. gondii in soil, targeting the repetitive 529 bp fragment of T. gondii genomic DNA. Then a total of 268 soil samples, including 148 samples from Gansu Province and 120 samples from Qinghai Province, northwestern China, were examined by the semi-nested PCR method. One third of the positive samples were sequenced. The sensitivity of the semi-nested PCR assay was 10(2)  T. gondii oocysts in 5 g soil sample. Investigation of soil samples from northwestern China showed that 34 out of 268 soil samples (12.69%) were T. gondii positive. Sequences of the partial 529 bp fragments varied from 0-1.2% among the sequenced samples. The prevalence of T. gondii oocysts in soil from cities (24/163) was slightly higher than that in soils from pasturing areas (10/105) (P = 0.21). Among the different regions in cities, the prevalence of T. gondii oocysts in soils from parks was 14.15%, whereas that in soils from schools was 19.05%. The present study firstly reported the prevalence of T. gondii oocysts in soils in northwest China using a novel semi-nested PCR assay, which provided baseline data for the effective prevention and control of toxoplasmosis in this region.

  15. Seroprevalence and Associated Risk Factors for Toxoplasma gondii Infection in Healthy Blood Donors: A Cross-Sectional Study in Sonora, Mexico.

    Science.gov (United States)

    Alvarado-Esquivel, Cosme; Rascón-Careaga, Antonio; Hernández-Tinoco, Jesús; Corella-Madueño, María Alba Guadalupe; Sánchez-Anguiano, Luis Francisco; Aldana-Madrid, María Lourdes; Velasquez-Vega, Edgar; Quizán-Plata, Trinidad; Navarro-Henze, José Luis; Badell-Luzardo, Joel Alberto; Gastélum-Cano, José María; Liesenfeld, Oliver

    2016-01-01

    Toxoplasma gondii (T. gondii) can be transmitted by blood transfusion. We determined the prevalence of T. gondii infection in healthy blood donors in Hermosillo city, Mexico, and the association of infection with T. gondii with the sociodemographic, clinical, and behavioral characteristics of blood donors. Four hundred and eight blood donors who attended two public blood banks in Hermosillo city were examined for anti-T. gondii IgG and IgM antibodies by using enzyme-linked immunoassays. Of the 408 blood donors (mean age 31.77 ± 9.52; range 18-60 years old) studied, 55 (13.5%) were positive for anti-T. gondii IgG antibodies, and 12 (21.8%) of them were also positive for anti-T. gondii IgM antibodies. Multivariate analysis showed that seropositivity to T. gondii was associated with age (OR = 1.74; 95% CI: 1.03-2.94; P = 0.03) and tobacco use (OR = 2.09; 95% CI: 1.02-4.29; P = 0.04). Seropositivity to T. gondii was correlated with the number of pregnancies, deliveries, and cesarean sections. The seroprevalence of T. gondii infection in blood donors in Sonora is the highest reported in blood donors in northern Mexico so far. This is the first report of an association of T. gondii exposure and tobacco use. Further research to confirm this association is needed.

  16. Pancreatic acinar cells-derived cyclophilin A promotes pancreatic damage by activating NF-κB pathway in experimental pancreatitis

    International Nuclear Information System (INIS)

    Yu, Ge; Wan, Rong; Hu, Yanling; Ni, Jianbo; Yin, Guojian; Xing, Miao; Shen, Jie; Tang, Maochun; Chen, Congying; Fan, Yuting; Xiao, Wenqin; Zhao, Yan; Wang, Xingpeng

    2014-01-01

    Highlights: • CypA is upregulated in experimental pancreatitis. • CCK induces expression and release of CypA in acinar cell in vitro. • rCypA aggravates CCK-induced acinar cell death and inflammatory cytokine production. • rCypA activates the NF-κB pathway in acinar cells in vitro. - Abstract: Inflammation triggered by necrotic acinar cells contributes to the pathophysiology of acute pancreatitis (AP), but its precise mechanism remains unclear. Recent studies have shown that Cyclophilin A (CypA) released from necrotic cells is involved in the pathogenesis of several inflammatory diseases. We therefore investigated the role of CypA in experimental AP induced by administration of sodium taurocholate (STC). CypA was markedly upregulated and widely expressed in disrupted acinar cells, infiltrated inflammatory cells, and tubular complexes. In vitro, it was released from damaged acinar cells by cholecystokinin (CCK) induction. rCypA (recombinant CypA) aggravated CCK-induced acinar cell necrosis, promoted nuclear factor (NF)-κB p65 activation, and increased cytokine production. In conclusion, CypA promotes pancreatic damage by upregulating expression of inflammatory cytokines of acinar cells via the NF-κB pathway

  17. CD147 stimulates hepatoma cells escaping from immune surveillance of T cells by interaction with Cyclophilin A.

    Science.gov (United States)

    Ren, Yi-Xin; Wang, Shu-Jing; Fan, Jian-Hui; Sun, Shi-Jie; Li, Xia; Padhiar, Arshad Ahmed; Zhang, Jia-Ning

    2016-05-01

    T cells play an important role in tumor immune surveillance. CD147 is a member of immunoglobulin superfamily present on the surface of many tumor cells and mediates malignant cell behaviors. Cyclophilin A (CypA) is an intracellular protein promoting inflammation when released from cells. CypA is a natural ligand for CD147. In this study, CD147 specific short hairpin RNAs (shRNA) were transfected into murine hepatocellular carcinoma Hepa1-6 cells to assess the effects of CD147 on hepatoma cells escaping from immune surveillance of T cells. We found extracellular CypA stimulated cell proliferation through CD147 by activating ERK1/2 signaling pathway. Downregulation of CD147 expression on Hepa1-6 cells significantly suppressed tumor progression in vivo, and decreased cell viability when co-cultured with T cells in vitro. Importantly, knockdown of CD147 on Hepa1-6 cells resulted in significantly increased T cells chemotaxis induced by CypA both in vivo and in vitro. These findings provide novel mechanisms how tumor cells escaping from immune surveillance of T cells. We provide a potential therapy for hepatocellular carcinoma by targeting CD147 or CD147-CypA interactions. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  18. Cyclophilin B induces chemoresistance by degrading wild type p53 via interaction with MDM2 in colorectal cancer.

    Science.gov (United States)

    Choi, Tae Gyu; Nguyen, Minh Nam; Kim, Jieun; Jo, Yong Hwa; Jang, Miran; Nguyen, Ngoc Ngo Yen; Yun, Hyeong Rok; Choe, Wonchae; Kang, Insug; Ha, Joohun; Tang, Dean G; Kim, Sung Soo

    2018-06-06

    Colorectal cancer (CRC) is one of the leading causes of cancer-related deaths worldwide. Chemoresistance is a major problem for effective therapy in CRC. Here, we investigated the mechanism by which peptidylprolyl isomerase B (PPIB; cyclophilin B, CypB) regulates chemoresistance in CRC. We found that CypB is a novel wild type p53 (p53WT)-inducible gene but a negative regulator of p53WT in response to oxaliplatin treatment. Overexpression of CypB shortens the half-life of p53WT and inhibits oxaliplatin-induced apoptosis in CRC cells, whereas knockdown of CypB lengthens the half-life of p53WT and stimulates p53WT dependent apoptosis. CypB interacts directly with MDM2, and enhances MDM2-dependent p53WT ubiquitination and degradation. Furthermore, we firmly validated using bioinformatics analyses that overexpression of CypB is associated with poor prognosis in CRC progression and chemoresistance. Hence, we suggest a novel mechanism of chemoresistance caused by overexpressed CypB, which may help to develop new anti-cancer drugs. We also propose that CypB may be utilized as a predictive biomarker in CRC patients. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  19. Inhibition of cyclophilin A suppresses H2O2-enhanced replication of HCMV through the p38 MAPK signaling pathway.

    Science.gov (United States)

    Xiao, Jun; Song, Xin; Deng, Jiang; Lv, Liping; Ma, Ping; Gao, Bo; Zhou, Xipeng; Zhang, Yanyu; Xu, Jinbo

    2016-09-01

    Human cytomegalovirus (HCMV) infection can be accelerated by intracellular and extracellular hydrogen peroxide (H2O2) stimulation, mediated by the activation of the p38 mitogen-activated protein kinase (MAPK) pathway. However, it remains unknown whether host gene expression is involved in H2O2-upregulated HCMV replication. Here, we show that the expression of the host gene, cyclophilin A (CyPA), could be facilitated by treatment with H2O2 in a dose-dependent manner. Experiments with CyPA-specific siRNA, or with cyclosporine A, an inhibitor of CyPA, confirmed that H2O2-mediated upregulation of HCMV replication is specifically mediated by upregulation of CyPA expression. Furthermore, depletion or inhibition of CyPA reduced H2O2-induced p38 activation, consistent with that of H2O2-upregulated HCMV lytic replication. These results show that H2O2 is capable of activating ROS-CyPA-p38 MAPK interactions to enhance HCMV replication.

  20. Pancreatic acinar cells-derived cyclophilin A promotes pancreatic damage by activating NF-κB pathway in experimental pancreatitis

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Ge [Department of Gastroenterology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai (China); Wan, Rong [Department of Gastroenterology, Shanghai First People’s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai (China); Hu, Yanling [Department of Gastroenterology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai (China); Ni, Jianbo [Department of Gastroenterology, Shanghai First People’s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai (China); Yin, Guojian; Xing, Miao [Department of Gastroenterology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai (China); Shen, Jie [Department of Gastroenterology, Shanghai First People’s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai (China); Tang, Maochun [Department of Gastroenterology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai (China); Chen, Congying [Department of Gastroenterology, Shanghai First People’s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai (China); Fan, Yuting; Xiao, Wenqin; Zhao, Yan [Department of Gastroenterology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai (China); Wang, Xingpeng, E-mail: wangxingpeng@hotmail.com [Department of Gastroenterology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai (China); Department of Gastroenterology, Shanghai First People’s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai (China); and others

    2014-01-31

    Highlights: • CypA is upregulated in experimental pancreatitis. • CCK induces expression and release of CypA in acinar cell in vitro. • rCypA aggravates CCK-induced acinar cell death and inflammatory cytokine production. • rCypA activates the NF-κB pathway in acinar cells in vitro. - Abstract: Inflammation triggered by necrotic acinar cells contributes to the pathophysiology of acute pancreatitis (AP), but its precise mechanism remains unclear. Recent studies have shown that Cyclophilin A (CypA) released from necrotic cells is involved in the pathogenesis of several inflammatory diseases. We therefore investigated the role of CypA in experimental AP induced by administration of sodium taurocholate (STC). CypA was markedly upregulated and widely expressed in disrupted acinar cells, infiltrated inflammatory cells, and tubular complexes. In vitro, it was released from damaged acinar cells by cholecystokinin (CCK) induction. rCypA (recombinant CypA) aggravated CCK-induced acinar cell necrosis, promoted nuclear factor (NF)-κB p65 activation, and increased cytokine production. In conclusion, CypA promotes pancreatic damage by upregulating expression of inflammatory cytokines of acinar cells via the NF-κB pathway.

  1. Study of the participation of MMP-7, EMMPRIN and cyclophilin A in the pathogenesis of periodontal disease.

    Science.gov (United States)

    de Oliveira Nóbrega, Fernando José; de Oliveira, Denise Hélen Imaculada Pereira; Vasconcelos, Rodrigo Gadelha; Nonaka, Cassiano Francisco Weege; Queiroz, Lélia Maria Guedes

    2016-12-01

    Periodontal disease is an infectious disease resulting from the immunoinflammatory response of the host to microorganisms present in the dental biofilm which causes tissue destruction. The objective of this study was to evaluate the immunohistochemical expression of matrix metalloproteinase 7 (MMP-7), extracellular matrix metalloproteinase inducer (EMMPRIN) and cyclophilin A (CypA) in periodontal disease. Gingival tissue samples were divided as follows: clinically healthy gingiva (n=32), biofilm-induced gingivitis (n=28), and chronic periodontitis (n=30). Histological sections of 3μm were submitted to immunoperoxidase method and undergone quantitative analysis. The results were analyzed statistically by the Mann-Whitney and Spearman correlation tests, with the level of significance set at 0.05 (α=0.05). Immunopositivity for MMP-7, EMMPRIN and CypA differed significantly between the three groups, with higher percentages of staining in chronic periodontitis specimens, followed by chronic gingivitis and healthy gingiva specimens (pEMMPRIN (r=0.289; p=0.006). In addition, there was a significant positive correlation between probing depth and expression of MMP-7 (r=0.726; pEMMPRIN (r=0.345; p=0.001), and CypA (r=0.803; pEMMPRIN and CypA are associated with the pathogenesis and progression of periodontal disease. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. Cyclophilin B is involved in p300-mediated degradation of CHOP in tumor cell adaptation to hypoxia.

    Science.gov (United States)

    Jeong, K; Kim, H; Kim, K; Kim, S-J; Hahn, B-S; Jahng, G-H; Yoon, K-S; Kim, S S; Ha, J; Kang, I; Choe, W

    2014-03-01

    The regulation of CCAAT/enhancer-binding protein-homologous protein (CHOP), an endoplasmic reticulum (ER) stress-response factor, is key to cellular survival. Hypoxia is a physiologically important stress that induces cell death in the context of the ER, especially in solid tumors. Although our previous studies have suggested that Cyclophilin B (CypB), a molecular chaperone, has a role in ER stress, currently, there is no direct information supporting its mechanism under hypoxia. Here, we demonstrate for the first time that CypB is associated with p300 E4 ligase, induces ubiquitination and regulates the proteasomal turnover of CHOP, one of the well-known pro-apoptotic molecules under hypoxia. Our findings show that CypB physically interacts with the N-terminal α-helix domain of CHOP under hypoxia and cooperates with p300 to modulate the ubiquitination of CHOP. We also show that CypB is transcriptionally induced through ATF6 under hypoxia. Collectively, these findings demonstrate that CypB prevents hypoxia-induced cell death through modulation of ubiquitin-mediated CHOP protein degradation, suggesting that CypB may have an important role in the tight regulation of CHOP under hypoxia.

  3. Cyclophilin B as a co-regulator of prolactin-induced gene expression and function in breast cancer cells.

    Science.gov (United States)

    Fang, Feng; Zheng, Jiamao; Galbaugh, Traci L; Fiorillo, Alyson A; Hjort, Elizabeth E; Zeng, Xianke; Clevenger, Charles V

    2010-06-01

    The effects of prolactin (PRL) during the pathogenesis of breast cancer are mediated in part though Stat5 activity enhanced by its interaction with its transcriptional inducer, the prolyl isomerase cyclophilin B (CypB). We have demonstrated that knockdown of CypB decreases cell growth, proliferation, and migration, and CypB expression is associated with malignant progression of breast cancer. In this study, we examined the effect of CypB knockdown on PRL signaling in breast cancer cells. CypB knockdown with two independent siRNAs was shown to impair PRL-induced reporter expression in breast cancer cell line. cDNA microarray analysis was performed on these cells to assess the effect of CypB reduction, and revealed a significant decrease in PRL-induced endogenous gene expression in two breast cancer cell lines. Parallel functional assays revealed corresponding alterations of both anchorage-independent cell growth and cell motility of breast cancer cells. Our results demonstrate that CypB expression levels significantly modulate PRL-induced function in breast cancer cells ultimately resulting in enhanced levels of PRL-responsive gene expression, cell growth, and migration. Given the increasingly appreciated role of PRL in the pathogenesis of breast cancer, the actions of CypB detailed here are of biological significance.

  4. Identification of cellular and viral factors related to anti-hepatitis C virus activity of cyclophilin inhibitor.

    Science.gov (United States)

    Goto, Kaku; Watashi, Koichi; Inoue, Daisuke; Hijikata, Makoto; Shimotohno, Kunitada

    2009-10-01

    We have so far reported that an immunosuppressant cyclosporin A (CsA), a well-known cyclophilin (CyP) inhibitor (CPI), strongly suppressed hepatitis C virus (HCV) replication in cell culture, and that CyPB was a cellular cofactor for viral replication. To further investigate antiviral mechanisms of CPI, we here developed cells carrying CsA-resistant HCV replicons, by culturing the HCV subgenomic replicon cells for 4 weeks in the presence of CsA with G418. Transfection of total RNA from the isolated CsA-resistant cells to naïve Huh7 cells conferred CsA resistance, suggesting that the replicon RNA itself was responsible for the resistant phenotype. Of the identified amino acid mutations, D320E in NS5A conferred the CsA resistance. The replicon carrying the D320E mutation was sensitive to interferon-alpha, but was resistant to CsA and other CPIs including NIM811 and sanglifehrin A. Knockdown of individual CyP subtypes revealed CyP40, in addition to CyPA and CyPB, contributed to viral replication, and CsA-resistant replicons acquired independence from CyPA for efficient replication. These data provide important evidence on the mechanisms underlying the regulation of HCV replication by CyP and for designing novel and specific anti-HCV strategies with CPIs.

  5. Comparing human pancreatic cell secretomes by in vitro aptamer selection identifies cyclophilin B as a candidate pancreatic cancer biomarker.

    Science.gov (United States)

    Ray, Partha; Rialon-Guevara, Kristy L; Veras, Emanuela; Sullenger, Bruce A; White, Rebekah R

    2012-05-01

    Most cases of pancreatic cancer are not diagnosed until they are no longer curable with surgery. Therefore, it is critical to develop a sensitive, preferably noninvasive, method for detecting the disease at an earlier stage. In order to identify biomarkers for pancreatic cancer, we devised an in vitro positive/negative selection strategy to identify RNA ligands (aptamers) that could detect structural differences between the secretomes of pancreatic cancer and non-cancerous cells. Using this molecular recognition approach, we identified an aptamer (M9-5) that differentially bound conditioned media from cancerous and non-cancerous human pancreatic cell lines. This aptamer further discriminated between the sera of pancreatic cancer patients and healthy volunteers with high sensitivity and specificity. We utilized biochemical purification methods and mass-spectrometric analysis to identify the M9-5 target as cyclophilin B (CypB). This molecular recognition-based strategy simultaneously identified CypB as a serum biomarker and generated a new reagent to recognize it in body fluids. Moreover, this approach should be generalizable to other diseases and complementary to traditional approaches that focus on differences in expression level between samples. Finally, we suggest that the aptamer we identified has the potential to serve as a tool for the early detection of pancreatic cancer.

  6. Ultra-violet B (UVB)-induced skin cell death occurs through a cyclophilin D intrinsic signaling pathway.

    Science.gov (United States)

    Ji, Chao; Yang, Bo; Yang, Zhi; Tu, Ying; Yang, Yan-li; He, Li; Bi, Zhi-Gang

    2012-09-07

    UVB-induced skin cell damage involves the opening of mitochondrial permeability transition pore (mPTP), which leads to both apoptotic and necrotic cell death. Cyclophilin D (Cyp-D) translocation to the inner membrane of mitochondrion acts as a key component to open the mPTP. Our Western-Blot results in primary cultured human skin keratinocytes and in HaCaT cell line demonstrated that UVB radiation and hydrogen peroxide (H(2)O(2)) induced Cyp-D expression, which was inhibited by anti-oxidant N-acetyl cysteine (NAC). We created a stable Cyp-D deficiency skin keratinocytes by expressing Cyp-D-shRNA through lentiviral infection. Cyp-D-deficient cells were significantly less susceptible than their counterparts to UVB- or H(2)O(2)-induced cell death. Further, cyclosporine A (Cs-A), a Cyp-D inhibitor, inhibited UVB- or H(2)O(2)-induced keratinocytes cell death. Reversely, over-expression of Cyp-D in primary keratinocytes caused spontaneous keratinocytes cell death. These results suggest Cyp-D's critical role in UVB/oxidative stress-induced skin cell death. Copyright © 2012 Elsevier Inc. All rights reserved.

  7. Molecular detection of Toxoplasma gondii infection in slaughtered ruminants (sheep, goats and cattle) in Northwest Tunisia.

    Science.gov (United States)

    Amdouni, Yosra; Rjeibi, Mohamed Ridha; Rouatbi, Mariem; Amairia, Safa; Awadi, Sofia; Gharbi, Mohamed

    2017-11-01

    The present study aimed to estimate the molecular prevalence of T. gondii infection in meat from slaughtered sheep, goats and cattle in Northwest Tunisia (Béja district). PCRs were performed on genomic DNA extracted from 420 meat samples (150 ewes, 120 goats and 150 cows). The overall molecular prevalence of T. gondii in sheep, goats and cattle were 33.3 (50/150), 32.5 (39/120) and 19.3% (29/150), respectively. Toxoplasma gondii molecular prevalences in the three meat ruminant species were significantly higher in adults compared to young animals (pgoats (pmeat. Extension programmes should be implemented to decrease the risk of infection related to sheep, goats and cattle meat manipulation and raw or undercooked meat consumption. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Seroprevalence of Toxoplasma gondii infection in zoo and domestic animals in Jiangxi Province, China

    Directory of Open Access Journals (Sweden)

    Luo Houqiang

    2017-01-01

    Full Text Available Toxoplasma gondii is a zoonotic protozoan parasite that infects a wide range of warm-blooded animals throughout the world. In the present study, antibodies to T. gondii were determined using a commercial indirect hemagglutination (IHA test in wild animals in a zoo. Three of 11 giraffes (Giraffa camelopardalis (27%, 1 of 5 wolves (Canis lupus laniger (20%, 1 of 6 hippopotamuses (Hippopotamus amphibious (17%, and 2 of 9 tundra swans (Cygnus columbianus (22% were found to be positive. No antibodies were detected in leopards (Panthera pardus, wild geese (Anser cygnoides, and Eastern grey kangaroos (Macropus giganteus. Domestic species from 13 counties of Jiangxi Province, China were also investigated by an indirect hemagglutination (IHA test. Thirty-five of 340 goats (10%, 94 of 560 water buffaloes (17%, and 4 of 35 cattle (11% were found to be seropositive. This is the first report of T. gondii infection in animals kept in zoos and domestic animals in this province.

  9. Seroprevalence of Toxoplasma gondii in free-ranging and feral cats on Amami Oshima Island, Japan.

    Science.gov (United States)

    Matsuu, Aya; Yokota, Shin-Ichi; Ito, Keiko; Masatani, Tatsunori

    2017-11-17

    On Amami Oshima Island, free-ranging and feral cats are harmful to wildlife populations. In this study, the seroprevalence of Toxoplasma gondii in these cats was examined using a newly developed Gaussia luciferase immunoprecipitation system assay. Of 1,363 cats, 123 cats (9.0%) was positive for T. gondii. The prevalence was significantly different in different areas; among cats in the rural area, where many wild animals live, including endangered species, T. gondii infection was more prevalent than in the urban area of the island. This finding indicates a possible risk to wildlife of infection from free-ranging and feral cats. Therefore, management of cats is important for wildlife conservation.

  10. Screening of chemical compound libraries identified new anti-Toxoplasma gondii agents.

    Science.gov (United States)

    Adeyemi, Oluyomi Stephen; Sugi, Tatsuki; Han, Yongmei; Kato, Kentaro

    2018-02-01

    Toxoplasma gondii is the etiological agent of toxoplasmosis, a common parasitic disease that affects nearly one-third of the human population. The primary infection can be asymptomatic in healthy individuals but may prove fatal in immunocompromised individuals. Available treatment options for toxoplasmosis patients are limited, underscoring the urgent need to identify and develop new therapies. Non-biased screening of libraries of chemical compounds including the repurposing of well-characterized compounds is emerging as viable approach to achieving this goal. In the present investigation, we screened libraries of natural product and FDA-approved compounds to identify those that inhibited T. gondii growth. We identified 32 new compounds that potently inhibit T. gondii growth. Our findings are new and promising, and further strengthen the prospects of drug repurposing as well as the screening of a wide range of chemical compounds as a viable source of alternative anti-parasitic therapeutic agents.

  11. ARF6, PI3-kinase and host cell actin cytoskeleton in Toxoplasma gondii cell invasion

    International Nuclear Information System (INIS)

    Vieira da Silva, Claudio; Alves da Silva, Erika; Costa Cruz, Mario; Chavrier, Philippe; Arruda Mortara, Renato

    2009-01-01

    Toxoplasma gondii infects a variety of different cell types in a range of different hosts. Host cell invasion by T. gondii occurs by active penetration of the host cell, a process previously described as independent of host actin polymerization. Also, the parasitophorous vacuole has been shown to resist fusion with endocytic and exocytic pathways of the host cell. ADP-ribosylation factor-6 (ARF6) belongs to the ARF family of small GTP-binding proteins. ARF6 regulates membrane trafficking and actin cytoskeleton rearrangements at the plasma membrane. Here, we have observed that ARF6 is recruited to the parasitophorous vacuole of tachyzoites of T. gondii RH strain and it also plays an important role in the parasite cell invasion with activation of PI3-kinase and recruitment of PIP 2 and PIP 3 to the parasitophorous vacuole of invading parasites. Moreover, it was verified that maintenance of host cell actin cytoskeleton integrity is important to parasite invasion.

  12. DNA chip-assisted diagnosis of a previously unknown etiology of intermediate uveitis- Toxoplasma gondii

    Directory of Open Access Journals (Sweden)

    Basu Soumyava

    2010-01-01

    Full Text Available We report the use of DNA chip technology in the identification of Toxoplasma gondii as the etiological agent in two patients with recurrent intermediate uveitis (IU. Both patients had recurrent episodes of vitritis (with no focal retinochoroidal lesion over varying time intervals and were diagnosed to have IU. The tuberculin test was negative in both. Blood counts, erythrocyte sedimentation rate, and serum angiotensin convertase enzyme levels were normal. In both cases, the vitreous fluid tested positive for the T. gondii DNA sequence by using a uveitis DNA chip (XCyton Pvt. Ltd., Bangalore, India. It contained complimentary sequences to "signature genes" of T. gondii, Mycobacterium tuberculosis, M. chelonae, and M. fortuitum. The enzyme-linked immunosorbent assay (ELISA detected elevated serum antitoxoplasma IgG levels in both. They responded to the antitoxoplasma therapy with oral co-trimoxazole (and additional intravitreal clindamycin in patient 1, with no recurrence during follow-ups of 6 and 8 months, respectively.

  13. Toxoplasma gondii infection reduces predator aversion in rats through epigenetic modulation in the host medial amygdala.

    Science.gov (United States)

    Hari Dass, Shantala Arundhati; Vyas, Ajai

    2014-12-01

    Male rats (Rattus novergicus) infected with protozoan Toxoplasma gondii relinquish their innate aversion to the cat odours. This behavioural change is postulated to increase transmission of the parasite to its definitive felid hosts. Here, we show that the Toxoplasma gondii infection institutes an epigenetic change in the DNA methylation of the arginine vasopressin promoter in the medial amygdala of male rats. Infected animals exhibit hypomethylation of arginine vasopressin promoter, leading to greater expression of this nonapeptide. The infection also results in the greater activation of the vasopressinergic neurons after exposure to the cat odour. Furthermore, we show that loss of fear in the infected animals can be rescued by the systemic hypermethylation and recapitulated by directed hypomethylation in the medial amygdala. These results demonstrate an epigenetic proximate mechanism underlying the extended phenotype in the Rattus novergicus-Toxoplasma gondii association. © 2014 John Wiley & Sons Ltd.

  14. A seroepidemiological survey of Toxoplasma gondii infection in referred dogs to Veterinary Hospital of Ahvaz, Iran

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    Fatemeh Zarra-Nezhad

    2017-12-01

    Full Text Available Toxoplasma gondii is an intracellular protozoan parasite, which is the cause of toxoplasmosis and can infect a wide variety of warm-blooded animals, including dogs and humans. The present study evaluated the seroprevalence of T. gondii infection in pet dogs in Ahvaz, southwest city of Iran, and investigated the related possible risk factors. A total of 180 serum samples were collected from dogs referred to Veterinary Hospital of Ahvaz. The samples were then tested by indirect enzyme-linked immunosorbent assay. The prevalence of T. gondii antibodies were 46.67%. Logistic regression and chi square tests were used for evaluating of risk factors. The positivity increased statistically significantly with dog’s gender (56% females and 39% males, P = 0.001, age (18% in <2 years old, 96% in ≥4 years old, P = 0.001 and place of living (47% outdoor dogs 38% house hold dogs, P = 0.025. However, no statistically significant association was found with dogs’ breed, deworming, food ingestion or contact with cats. Overall, the results showed a relatively high seroprevalence of T. gondii infection in dogs in southwest Iran (Ahvaz and proved association of T. gondii prevalence rates with the dog’s age, gender and place of living. Keywords: Toxoplasma gondii, Toxoplasmosis, Indirect enzyme-linked immunosorbent assay, Outdoor dogs, House hold dogs, Deworming

  15. toxoMine: an integrated omics data warehouse for Toxoplasma gondii systems biology research.

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    Rhee, David B; Croken, Matthew McKnight; Shieh, Kevin R; Sullivan, Julie; Micklem, Gos; Kim, Kami; Golden, Aaron

    2015-01-01

    Toxoplasma gondii (T. gondii) is an obligate intracellular parasite that must monitor for changes in the host environment and respond accordingly; however, it is still not fully known which genetic or epigenetic factors are involved in regulating virulence traits of T. gondii. There are on-going efforts to elucidate the mechanisms regulating the stage transition process via the application of high-throughput epigenomics, genomics and proteomics techniques. Given the range of experimental conditions and the typical yield from such high-throughput techniques, a new challenge arises: how to effectively collect, organize and disseminate the generated data for subsequent data analysis. Here, we describe toxoMine, which provides a powerful interface to support sophisticated integrative exploration of high-throughput experimental data and metadata, providing researchers with a more tractable means toward understanding how genetic and/or epigenetic factors play a coordinated role in determining pathogenicity of T. gondii. As a data warehouse, toxoMine allows integration of high-throughput data sets with public T. gondii data. toxoMine is also able to execute complex queries involving multiple data sets with straightforward user interaction. Furthermore, toxoMine allows users to define their own parameters during the search process that gives users near-limitless search and query capabilities. The interoperability feature also allows users to query and examine data available in other InterMine systems, which would effectively augment the search scope beyond what is available to toxoMine. toxoMine complements the major community database ToxoDB by providing a data warehouse that enables more extensive integrative studies for T. gondii. Given all these factors, we believe it will become an indispensable resource to the greater infectious disease research community. © The Author(s) 2015. Published by Oxford University Press.

  16. Age-related Toxoplasma gondii seroprevalence in Dutch wild boar inconsistent with lifelong persistence of antibodies.

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    Marieke Opsteegh

    Full Text Available Toxoplasma gondii is an important zoonotic pathogen that is best known as a cause of abortion or abnormalities in the newborn after primary infection during pregnancy. Our aim was to determine the prevalence of T. gondii in wild boar to investigate the possible role of their meat in human infection and to get an indication of the environmental contamination with T. gondii. The presence of anti-T. gondii antibodies was determined by in-house ELISA in 509 wild boar shot in 2002/2003 and 464 wild boar shot in 2007. Most of the boar originated from the "Roerstreek" (n = 673 or the "Veluwe" (n = 241. A binormal mixture model was fitted to the log-transformed optical density values for wild boar up to 20 months old to estimate the optimal cut-off value (-0.685 and accompanying sensitivity (90.6% and specificity (93.6%. The overall seroprevalence was estimated at 24.4% (95% CI: 21.1-27.7%. The prevalence did not show variation between sampling years or regions, indicating a stable and homogeneous infection pressure from the environment. The relation between age and seroprevalence was studied in two stages. Firstly, seroprevalence by age group was determined by fitting the binary mixture model to 200 animals per age category. The prevalence showed a steep increase until approximately 10 months of age but stabilized at approximately 35% thereafter. Secondly, we fitted the age-dependent seroprevalence data to several SIR-type models, with seropositives as infected (I and seronegatives as either susceptible (S or resistant (R. A model with a recovery rate (SIS was superior to a model without a recovery rate (SI. This finding is not consistent with the traditional view of lifelong persistence of T. gondii infections. The high seroprevalence suggests that eating undercooked wild boar meat may pose a risk of infection with T. gondii.

  17. Comparison of three diagnostic methods for the detection of Toxoplasma gondii in free range chickens.

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    Hamidinejat, H; Nabavi, L; Mayahi, M; Ghourbanpoor, M; Pourmehdi Borojeni, M; Norollahi Fard, S; Shokrollahi, M

    2014-09-01

    Detection of Toxoplasma gondii in free range chickens is an indicator of the prevalence and distribution pattern of T. gondii in the environment. For this purpose, serologic assays especially modified agglutination test (MAT) is the main approach in the literature. The main goal of this study was to compare the polymerase chain reaction (PCR) based on amplification of first internal transcribed spacer (ITS-1) of ribosomal DNA gene, ELISA, and MAT to demonstrate T. gondii infection in free range chicken. A total of 106 adult free - range chickens were killed. Blood, whole heart and brain samples were taken. Sera were examined for the presence of T. gondii antibodies by ELISA and MAT as well. Selected tissues were used for PCR and bioassay in mice. The results revealed that 48.11%, 51.89%, 46.23% and 27.36% of chickens were positive in ELISA, MAT, PCR and bioassay in mice respectively. Good correlation between the results of PCR, ELISA and MAT were detected, but not with bioassay in mice. Compared with PCR, the sensitivity and specificity of ELISA were 92.16% and 96.36% respectively and also for MAT, the sensitivity was 81.81% and the specificity was 92.15%. The specific diagnosis of T. gondii infection in chickens is central to a better understanding of the epidemiology and dynamics of transmission among the various host population and is particularly important for planning effective optimal prevention and control programs. Our data in the present study demonstrated that PCR, ELISA and the MAT are helpful and precise methods to detect T. gondii in naturally infected free-range chickens.

  18. Anti-gluten immune response following Toxoplasma gondii infection in mice.

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    Emily G Severance

    Full Text Available Gluten sensitivity may affect disease pathogenesis in a subset of individuals who have schizophrenia, bipolar disorder or autism. Exposure to Toxoplasma gondii is a known risk factor for the development of schizophrenia, presumably through a direct pathological effect of the parasite on brain and behavior. A co-association of antibodies to wheat gluten and to T. gondii in individuals with schizophrenia was recently uncovered, suggesting a coordinated gastrointestinal means by which T. gondii and dietary gluten might generate an immune response. Here, we evaluated the connection between these infectious- and food-based antigens in mouse models. BALB/c mice receiving a standard wheat-based rodent chow were infected with T. gondii via intraperitoneal, peroral and prenatal exposure methods. Significant increases in the levels of anti-gluten IgG were documented in all infected mice and in offspring from chronically infected dams compared to uninfected controls (repetitive measures ANOVAs, two-tailed t-tests, all p≤0.00001. Activation of the complement system accompanied this immune response (p≤0.002-0.00001. Perorally-infected females showed higher levels of anti-gluten IgG than males (p≤0.009 indicating that T. gondii-generated gastrointestinal infection led to a significant anti-gluten immune response in a sex-dependent manner. These findings support a gastrointestinal basis by which two risk factors for schizophrenia, T. gondii infection and sensitivity to dietary gluten, might be connected to produce the immune activation that is becoming an increasingly recognized pathology of psychiatric disorders.

  19. Toxoplasma gondii Infection Is Associated with Mitochondrial Dysfunction in-Vitro

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    Genevieve Syn

    2017-12-01

    Full Text Available Upon invasion of host cells, the ubiquitous pathogen Toxoplasma gondii manipulates several host processes, including re-organization of host organelles, to create a replicative niche. Host mitochondrial association to T. gondii parasitophorous vacuoles is rapid and has roles in modulating host immune responses. Here gene expression profiling of T. gondii infected cells reveals enrichment of genes involved in oxidative phosphorylation (OXPHOS and mitochondrial dysfunction 6 h post-infection. We identified 11 hub genes (HIF-1α, CASP8, FN1, POU5F1, CD44, ISG15, HNRNPA1, MDM2, RPL35, VHL, and NUPR1 and 10 predicted upstream regulators, including 4 endogenous regulators RICTOR, KDM5A, RB1, and D-glucose. We characterized a number of mitochondrial parameters in T. gondii infected human foreskin fibroblast cells over a 36 h time-course. In addition to the usual rapid recruitment and apparent enlargement of mitochondria around the parasitophorous vacuole we observed fragmented host mitochondria in infected cells, not linked to cellular apoptosis, from 24 h post-infection. An increase in mitochondrial superoxide levels in T. gondii infected cells was observed that required active parasite invasion and peaked at 30 h post-infection. Measurement of OXPHOS proteins showed decreased expression of Complex IV in infected cells at 24 h post-infection, followed by decreased expression of Complexes I and II at 36 h post-infection. No change occurred in Complex V. No difference in host mitochondrial membrane potential between infected and mock-infected cells was observed at any time. Our results show perturbation of host mitochondrial function following T. gondii infection that likely impacts on pathogenesis of disease.

  20. A unique dual activity amino acid hydroxylase in Toxoplasma gondii.

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    Elizabeth A Gaskell

    Full Text Available The genome of the protozoan parasite Toxoplasma gondii was found to contain two genes encoding tyrosine hydroxylase; that produces L-DOPA. The encoded enzymes metabolize phenylalanine as well as tyrosine with substrate preference for tyrosine. Thus the enzymes catabolize phenylalanine to tyrosine and tyrosine to L-DOPA. The catalytic domain descriptive of this class of enzymes is conserved with the parasite enzyme and exhibits similar kinetic properties to metazoan tyrosine hydroxylases, but contains a unique N-terminal extension with a signal sequence motif. One of the genes, TgAaaH1, is constitutively expressed while the other gene, TgAaaH2, is induced during formation of the bradyzoites of the cyst stages of the life cycle. This is the first description of an aromatic amino acid hydroxylase in an apicomplexan parasite. Extensive searching of apicomplexan genome sequences revealed an ortholog in Neospora caninum but not in Eimeria, Cryptosporidium, Theileria, or Plasmodium. Possible role(s of these bi-functional enzymes during host infection are discussed.

  1. Cancer in the parasitic protozoans Trypanosoma brucei and Toxoplasma gondii.

    Science.gov (United States)

    Lun, Zhao-Rong; Lai, De-Hua; Wen, Yan-Zi; Zheng, Ling-Ling; Shen, Ji-Long; Yang, Ting-Bo; Zhou, Wen-Liang; Qu, Liang-Hu; Hide, Geoff; Ayala, Francisco J

    2015-07-21

    Cancer is a general name for more than 100 malignant diseases. It is postulated that all cancers start from a single abnormal cell that grows out of control. Untreated cancers can cause serious consequences and deaths. Great progress has been made in cancer research that has significantly improved our knowledge and understanding of the nature and mechanisms of the disease, but the origins of cancer are far from being well understood due to the limitations of suitable model systems and to the complexities of the disease. In view of the fact that cancers are found in various species of vertebrates and other metazoa, here, we suggest that cancer also occurs in parasitic protozoans such as Trypanosoma brucei, a blood parasite, and Toxoplasma gondii, an obligate intracellular pathogen. Without treatment, these protozoan cancers may cause severe disease and death in mammals, including humans. The simpler genomes of these single-cell organisms, in combination with their complex life cycles and fascinating life cycle differentiation processes, may help us to better understand the origins of cancers and, in particular, leukemias.

  2. Toxoplasma gondii and schizophrenia: a review of published RCTs.

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    Chorlton, Sam D

    2017-07-01

    Over the last 60 years, accumulating evidence has suggested that acute, chronic, and maternal Toxoplasma gondii infections predispose to schizophrenia. More recent evidence suggests that chronically infected patients with schizophrenia present with more severe disease. After acute infection, parasites form walled cysts in the brain, leading to lifelong chronic infection and drug resistance to commonly used antiparasitics. Chronic infection is the most studied and closely linked with development and severity of schizophrenia. There are currently four published randomized controlled trials evaluating antiparasitic drugs, specifically azithromycin, trimethoprim, artemisinin, and artemether, in patients with schizophrenia. No trials have demonstrated a change in psychopathology with adjunctive treatment. Published trials have either selected drugs without evidence against chronic infection or used them at doses too low to reduce brain cyst burden. Furthermore, trials have failed to achieve sufficient power or account for confounders such as previous antipsychotic treatment, sex, age, or rhesus status on antiparasitic effect. There are currently no ongoing trials of anti-Toxoplasma therapy in schizophrenia despite ample evidence to justify further testing.

  3. Pathology, clinical signs, and tissue distribution of Toxoplasma gondii in experimentally infected reindeer (Rangifer tarandus

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    Émilie Bouchard

    2017-12-01

    Full Text Available Toxoplasma gondii is a zoonotic parasite found in vertebrates worldwide for which felids serve as definitive hosts. Despite low densities of felids in northern Canada, Inuit people in some regions show unexpectedly high levels of exposure, possibly through handling and consumption of Arctic wildlife. Free-ranging caribou (Rangifer tarandus are widely harvested for food across the Canadian North, show evidence of seroexposure to T. gondii, and are currently declining in numbers throughout the Arctic. We experimentally infected three captive reindeer (conspecific with caribou with 1000, 5000 or 10,000 oocysts of T. gondii via stomach intubation to assess clinical signs of infection, pathology, and tissue distribution. An unexposed reindeer served as a negative control. Signs of stress, aggression, and depression were noted for the first two weeks following infection. By 4 weeks post infection, all infected reindeer were positive on a modified agglutination test at the highest titer tested (1:200 for antibodies to T. gondii. At 20 weeks post infection, no gross abnormalities were observed on necropsy. Following histopathology and immunohistochemistry, tissue cysts were visualized in the reindeer given the highest and lowest dose of oocysts. Focal pleuritis and alveolitis were associated with respiratory problems in reindeer given the middle dose. DNA of T. gondii was detected following traditional DNA extraction and conventional PCR on 25 mg samples from 17/33 muscles and organs, and by magnetic capture DNA extraction from 100 g samples from all 26 tissues examined. This research demonstrated that reindeer/caribou can serve as intermediate hosts for T. gondii, and that the parasite may be associated with health effects in wildlife. The presence of T. gondii in all tissues tested, many of which are commonly consumed raw, smoked, or dried in northern communities, suggests that caribou may serve as a source of human exposure to T. gondii

  4. Prevalence of Toxoplasma gondii infections in Pennsylvania black bears, Ursus americanus.

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    Briscoe, N; Humphreys, J G; Dubey, J P

    1993-10-01

    Serum samples from 665 hunter-killed black bears killed in 1989 to 1992 throughout Pennsylvania (USA) were tested for Toxoplasma gondii antibodies by the agglutination test in dilutions of 1:25, 1:50, and 1:500. Toxoplasma gondii antibodies were found in 535 of 665 (80%) bears. Considering the highest dilutions at which antibodies were detected, prevalences were 10% at 1:25, 37% at 1:50 and 33% at 1:500. No significant difference in antibody prevalence was found between males (79%) and females (80%), but a significant difference was found between juvenile (65%) and adult (83%) bears.

  5. Seroprevalence of Toxoplasma gondii and Trichinella spiralis in North Carolina black bears (Ursus americanus).

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    Nutter, F B; Levine, J F; Stoskopf, M K; Gamble, H R; Dubey, J P

    1998-10-01

    Serum samples from 143 hunter-killed black bears were collected during the 1996 and 1997 black bear hunting seasons in eastern North Carolina. All samples were tested for antibodies to Toxoplasma gondii by the modified agglutination test. Antibodies to T. gondii were present in 120 of 143 (84%) bears. Females had significantly higher titers than males (Wilcoxon rank sums test, P = 0.045), and titers increased with age (Jonckheere test, P = 0.01). Samples collected during 1996 (n = 79) were tested for antibodies to Trichinella spiralis by enzyme-linked immunosorbent assay. No samples were positive for antibodies to T. spiralis.

  6. Seroepidemiology of Toxoplasma gondii in dogs in the State of Rio de Janeiro

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    Nathalie Costa da Cunha

    2016-11-01

    Full Text Available ABSTRACT. Cunha N.C., Cordeiro M.D., Bravo S.A.C., Matos P.C.M., Almosny N.R.P. & da Fonseca A.H. [Seroepidemiology of Toxoplasma gondii in dogs in the State of Rio de Janeiro.] Soroepidemiologia de Toxoplasma gondii em cães no estado do Rio de Janeiro. Revista Brasileira de Medicina Veterinária 38(supl. 3: 114-121, 2016. Departamento de Saúde Coletiva Veterinária e Saúde Pública, Faculdade de Veterinária. Universidade Federal Fluminense, Vital Brazil, Niterói, RJ, Brazil. E-mail: nathaliecunha@id.uff.br Toxoplasmosis is a serious public health problem worldwide as it can cause prenatal and neonatal morbidity and mortality in humans. Although dogs are not definitive hosts of T. gondii, they play an important role in the mechanical dissemination of oocysts. This study aimed to carry out a seroepidemiological investigation of anti-Toxoplasma gondii antibodies in domestic dogs from seven municipalities in the state of Rio de Janeiro, Brazil. A crosssectional epidemiological study was carried out to evaluate the profile of anti-Toxoplasma gondii IgG antibodies in dogs from canine sera from different municipalities in the state of Rio de Janeiro. The municipalities studied were Cachoeiras de Macacu, Guapimirim, Itaboraí, Magé, Resende, Seropédica and Silva Jardim. The detection of antibodies of the IgG class anti-Toxoplasma gondii was performed using the indirect enzyme immunoadsorption (ELISA assay and the statistical analyzes used were the chi-square test and the prevalence ratio. Of the 651 samples tested, 300 were reactive for T. gondii, representing a relative frequency of 46.08% of seroreactive dogs. It was concluded that dogs are good sentinels for evaluations of risk for occurrence of T. gondii, emphasizing those coming from rural areas and that there was no difference in the occurrence of serorreative dogs in front of different municipalities studies of the state of Rio de Janeiro.

  7. Prevalence of Toxoplasma gondii in small mammals from the Ardennes region, France.

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    Afonso, Eve; Poulle, Marie-Lazarine; Lemoine, Mélissa; Villena, Isabelle; Aubert, Dominique; Gilot-Fromont, Emmanuelle

    2007-11-01

    Serum samples from 218 small mammals trapped in forest and grassland in the Ardennes region (North-eastern France) were tested for antibodies to Toxoplasma gondii. Using the modified agglutination test, positive results were found in 4/92 Apodemus sp., 3/64 Clethrionomys glareolus, 0/26 Microtus agrestis, 0/4 Micromys minutus, 3/5 Sorex sp., 2/9 Arvicola terrestris, and 7/18 Talpa europaea. Toxoplasma gondii was not isolated from the heart of seropositive individuals after bioassay in mice. Seroprevalence was significantly higher in large fossorial mammals living in grassland than in small forest mammals, probably related to ecological factors.

  8. Toxoplasma gondii seroprevalence and genotype diversity in select wildlife species from the southeastern United States

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    Richard W. Gerhold

    2017-10-01

    Full Text Available Abstract Background Toxoplasma gondii is a widespread protozoan parasite that infects humans and other animals. Previous studies indicate some genotypes of T. gondii are more frequently isolated in wildlife than agricultural animals, suggesting a wild/feral animal diversity model. To determine seroprevalence and genetic diversity of T. gondii in southeastern US wildlife, we collected sera from 471 wild animals, including 453 mammals and 18 birds, between 2011 and 2014. These serum samples were assayed for T. gondii infection using the modified agglutination test (MAT. Heart or tongue tissues from 66 seropositive animals were bioassayed in mice and 19 isolates were obtained. The isolated parasites were genotyped by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP method employing 10 genetic markers. Results One hundred and ninety-six of 471 samples (41.6% had a titer ≥1:32 and were considered positive for T. gondii infection. Of 453 mammals, 195 (43% were seropositive, whereas only one (5.6% of 18 birds was seropositive. The seroprevalence in mammals was significantly higher than in the birds. Mammalian hosts with adequate samples size (≥ 20 comprised white-tailed deer (n = 241, feral hogs (n = 100, raccoons (n = 34 and coyotes (n = 22, with seroprevalences of 41.0%, 51.0%, 50.0% and 72.7%, respectively. Coyotes had significantly higher seroprevalence than the white-tailed deer. Genotyping revealed five distinct genotypes, including the ToxoDB PCR-RFLP genotype #5 (a.k.a type 12 for 15 isolates, genotype #3 (a.k.a. type II for 1 isolate, and genotypes #154, #167 and #216, each for 1 isolate. The results showed moderate to high infection rates of T. gondii in white-tailed deer, feral hogs, raccoons and coyotes. Genotyping results indicated limited genetic diversity and a dominance of genotype #5, which has been reported as a major type in wildlife in North America. Conclusions We conclude that T. gondii

  9. Carne de consumo e risco de transmissão de Toxoplasma gondii

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    Rosado, Joana de Carvalho Serra

    2009-01-01

    O consumo de carne de suíno (Sus domesticus) é considerado o maior factor de risco para a transmissão da toxoplasmose aos humanos. No entanto, apesar dos suínos serem muito usados na gastronomia portuguesa, tanto pelo consumo de carne, como pelo de enchidos, pouco se sabe sobre a prevalência e caracterização genética de Toxoplasma gondii nestes produtos alimentares, no país. Neste trabalho, pretendeu-se determinar a seroprevalência de anticorpos IgG anti-T. gondii em suínos de ...

  10. TAL effectors target the C-terminal domain of RNA polymerase II (CTD by inhibiting the prolyl-isomerase activity of a CTD-associated cyclophilin.

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    Mariane Noronha Domingues

    Full Text Available Transcriptional activator-like (TAL effectors of plant pathogenic bacteria function as transcription factors in plant cells. However, how TAL effectors control transcription in the host is presently unknown. Previously, we showed that TAL effectors of the citrus canker pathogen Xanthomonas citri, named PthAs, targeted the citrus protein complex comprising the thioredoxin CsTdx, ubiquitin-conjugating enzymes CsUev/Ubc13 and cyclophilin CsCyp. Here we show that CsCyp complements the function of Cpr1 and Ess1, two yeast cyclophilins that regulate transcription by the isomerization of proline residues of the regulatory C-terminal domain (CTD of RNA polymerase II. We also demonstrate that CsCyp, CsTdx, CsUev and four PthA variants interact with the citrus CTD and that CsCyp co-immunoprecipitate with the CTD in citrus cell extracts and with PthA2 transiently expressed in sweet orange epicotyls. The interactions of CsCyp with the CTD and PthA2 were inhibited by cyclosporin A (CsA, a cyclophilin inhibitor. Moreover, we present evidence that PthA2 inhibits the peptidyl-prolyl cis-trans isomerase (PPIase activity of CsCyp in a similar fashion as CsA, and that silencing of CsCyp, as well as treatments with CsA, enhance canker lesions in X. citri-infected leaves. Given that CsCyp appears to function as a negative regulator of cell growth and that Ess1 negatively regulates transcription elongation in yeast, we propose that PthAs activate host transcription by inhibiting the PPIase activity of CsCyp on the CTD.

  11. [The seroprevalence of Toxoplasma gondii in women from Sanliurfa, a province with a high raw meatball consumption].

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    Tekay, Fikret; Ozbek, Erdal

    2007-01-01

    Toxoplasma gondii is an obligate intracellular protozoan that can infect all kind of birds and all mammals including humans and is common throughout the world. The prevalence varies according to social and cultural habits, pet cats in homes and geographic factors. Domestic cats are considered to be an important source of Toxoplasma gondii infection. From January to June 2006, the prevalence of toxoplasmosis was retrospectively monitored from blood samples that had been sent to our laboratory in order to determine the levels of IgM and IgG. All the subjects were women and 2,586 blood samples were investigated with the chemiluminescence immunoassay method. The rates of Toxoplasma gondii IgM antibodies were found to be 3.0% (78/2,586) and that of Toxoplasma gondii IgG antibodies, 69.5% (1.798/2,586). The total rate of positivity of Toxoplasma gondii antibodies was 69.6% (1,801/2,586) and the negativity, 30.4% (785/2,586). The highest positive rates have been reported in the southeastern region of Turkey and the 69.6% detected in our study seems to be the highest rate. Raw meatball consumption is common in our region and raw meat has a high risk of Toxoplasma gondii infection by direct ingestion of tissue cysts. As a result we consider that the high frequency of Toxoplasma gondii seropositivity in this region is due to raw meatball consumption.

  12. Toxoplasma gondii seroprevalence and association with equine protozoal myeloencephalitis: A case-control study of Californian horses.

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    James, K E; Smith, W A; Packham, A E; Conrad, P A; Pusterla, N

    2017-06-01

    While toxoplasmosis is not commonly considered a clinical disease of equines, previous seroprevalence studies have reported differing background rates of Toxoplasma gondii infection in horses globally. The objective of this study was to evaluate possible associations between T. gondii seroprevalence and clinical signs of equine protozoal myeloencephalitis (EPM) in horses. Using a case-control study design, 720 Californian horses with neurologic signs compatible with EPM were compared to healthy, non-neurologic horses for the presence of T. gondii antibodies (using indirect fluorescent antibody tests [IFAT]). Toxoplasma gondii seroprevalence among cases and controls was determined at standard serum cut-offs: 40, 80, 160, 320, and 640. At a T. gondii titre cut-off of 320, horses with clinical signs compatible with EPM had 3.55 times the odds of a seropositive test compared to those without clinical signs (Pequines. Serologic testing of cerebrospinal fluid and isolation of T. gondii in EPM suspect cases should be considered. Future studies investigating the relationship between T. gondii and EPM are warranted. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Seroprevalence and risk factors of Neospora spp. and Toxoplasma gondii infections among horses and donkeys in Nigeria, West Africa.

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    Bártová, Eva; Sedlák, Kamil; Kobédová, Kateřina; Budíková, Marie; Joel Atuman, Yakubu; Kamani, Joshua

    2017-09-26

    Neospora spp. and Toxoplasma gondii are considered to be a globally distributed parasites affecting wide range of warm-blooded animals. Neosporosis has caused clinical illness in horses and consumption of horse meat has been epidemiologically linked to clinical toxoplasmosis in humans. This study was conducted to determine Neospora spp. and T. gondii antibodies and risk factors of infection in horses and donkeys from three states of Nigeria. A total of 144 samples were collected from clinically healthy animals (120 horses and 24 donkeys). The sera were tested for antibodies to Neospora spp. and T. gondii by indirect fluorescence antibody test, a titer ≥ 50 was considered positive. Seroprevalence data were statistically analyzed, considering the variables of gender, age, use, state, origin of breed and type of management. Antibodies to Neospora spp. and T. gondii were detected in 8% horses with titers 50 and in 24% horses with titers 50-800, respectively. Co-infection of both parasites was proved in three horses (3%). Statistical differences were found only for T. gondii seroprevalence in horses with different use, locality, origin and management (p-value ≤ 0.05). Antibodies to T. gondii were detected in four (17%) of 24 donkeys with statistical difference (p-value ≤ 0.05) in animals of different use; antibodies to Neospora spp. were not proved in any of the donkeys. This is the first seroprevalence study of Neospora spp. and T. gondii in equids from Nigeria.

  14. Seroprevalence of Toxoplasma gondii Infection in Patients of Intensive Care Unit in China: A Hospital Based Study

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    Yong-Biao Zhang

    2015-01-01

    Full Text Available The objective of this study was to estimate the seroprevalence of Toxoplasma gondii infection in 394 patients of intensive care unit (ICU in a hospital between April 2010 and March 2012 and analyze the association between T. gondii infection and ICU patients according to the species of disease. Toxoplasma serology was evaluated by ELISA method using a commercially available kit. Data of patients were obtained from the patients, informants, and medical examination records. Seventy-four (18.78% of 394 patients were positive for anti-T. gondii IgG antibodies demonstrating latent infection. Of these, the highest T. gondii seroprevalence was found in the age group of 31–45 years (27.45%, and the lowest was found in the age group of <30 years (12.5%. In addition, females (21.6% had a higher seroprevalence than males (18.36%. With respect to the species of disease, the patients with kidney diseases (57.14%, lung diseases (27.84%, and brain diseases (24% had high T. gondii seroprevalence. The present study represents the first survey of T. gondii seroprevalence in ICU patients in China, revealing an 18.78% seropositivity. Considering the particularities of ICU patients, molecular identification, genetic characterization, and diagnosis of T. gondii should be considered in future study.

  15. Molecular and serological prevalence of Toxoplasma gondii and Anaplasma spp. infection in goats from Chongqing Municipality, China

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    Zhou Zuoyong

    2018-01-01

    Full Text Available Toxoplasmosis and anaplasmosis are severe zoonotic diseases, the former caused by Toxoplasma gondii and the latter by Anaplasma spp. In the present study, 332 goat blood samples were randomly collected from Chongqing Municipality, China to screen for T. gondii and Anaplasma spp. We used a polymerase chain reaction (PCR to detect DNA, and enzyme-linked immunosorbent assay (ELISA to test for T. gondii antibodies. The prevalence of T. gondii and Anaplasma spp. was 38% and 35% respectively by PCR, and 42% for T. gondii antibodies by ELISA. The co-infection rate by T. gondii and Anaplasma was 13%, where the two predominant pathogens co-infecting were Anaplasma phagocytophilum + A. bovis (10%, followed by T. gondii + A. phagocytophilum (9.64%. While co-infection by three pathogens varied ranging from 1.81% to 5.72%, less than 1% of goats were found to be positive for four pathogens. This is the first investigation of T. gondii and Anaplasma spp. infection in goats from Chongqing.

  16. Incidence of adult brain cancers is higher in countries where the protozoan parasite Toxoplasma gondii is common

    Science.gov (United States)

    Thomas, Frédéric; Lafferty, Kevin D.; Brodeur, Jacques; Elguero, Eric; Gauthier-Clerc, Michel; Missé, Dorothée

    2012-01-01

    We explored associations between the common protozoan parasite Toxoplasma gondii and brain cancers in human populations. We predicted that T. gondii could increase the risk of brain cancer because it is a long-lived parasite that encysts in the brain, where it provokes inflammation and inhibits apoptosis. We used a medical geography approach based on the national incidence of brain cancers and seroprevalence of T. gondii. We corrected reports of incidence for national gross domestic product because wealth probably increases the ability to detect cancer. We also included gender, cell phone use and latitude as variables in our initial models. Prevalence of T. gondii explained 19 per cent of the residual variance in brain cancer incidence after controlling for the positive effects of gross domestic product and latitude among nations. Infection with T. gondii was associated with a 1.8-fold increase in the risk of brain cancers across the range of T. gondii prevalence in our dataset (4–67%). These results, though correlational, suggest that T. gondii should be investigated further as a possible oncogenic pathogen of humans.

  17. First report of Toxoplasma gondii infection in market-sold adult chickens, ducks and pigeons in northwest China

    Directory of Open Access Journals (Sweden)

    Cong Wei

    2012-06-01

    Full Text Available Abstract Background Toxoplasma gondii infection is a global concern, affecting a wide range of warm-blooded animals and humans worldwide, including poultry. Domestic and companion birds are considered to play an important role in the transmission of T. gondii to humans and other animals. However, little information on T. gondii infection in domestic birds in Lanzhou, northwest China was available. Therefore, this study was performed to determine the seroprevalence of T. gondii infection in domestic birds in Lanzhou, northwest China. Methods In the present study, the seroprevalence of T. gondii antibodies in 413 (305 caged and 108 free-range adult chickens, 334 (111 caged and 223 free-range adult ducks and 312 adult pigeons in Lanzhou, northwest China, were examined using the modified agglutination test (MAT. Results 30 (7.26% chickens, 38 (11.38% ducks and 37 (11.86% pigeons were found to be positive for T. gondii antibodies at the cut-off of 1:5. The prevalences in caged and free-range chickens were 6.23% and 10.19% respectively, however, statistical analysis showed that the difference was not significant (P > 0.05. The seroprevalences in caged and free-range ducks were 6.31% and 13.90% respectively, but the difference was not statistically significant (P > 0.05. Conclusions The results of the present survey indicated the presence of T. gondii infection in adult chickens, ducks and pigeons sold for meat in poultry markets in Lanzhou, northwest China, which poses a potential risk for T. gondii infection in humans and other animals in this region. This is the first seroprevalence study of T. gondii infection in domestic birds in this region.

  18. Toxoplasma gondii in experimentally infected Bos taurus and Bos indicus semen and tissues Toxoplasma gondii em semen e tecidos de Bos taurus and Bos indicus experimentalmente infectados

    Directory of Open Access Journals (Sweden)

    Leslie Scarpelli

    2009-01-01

    Full Text Available Eighteen young steers were inoculated with Toxoplasma gondii and randomly distributed into three groups of six animals each: GI, 2.5x10(5 "P" strain oocysts, GII, 5.0x10(6 "RH" strain tachyzoites, and GIII (Control. Clinical, serological and parasitemia exams were realized. Parasite investigation by bioassay and PCR was realized on semen and fragments of skeletal musculature, lymph nodes, brain, retina, spleen, liver, lung, testicle, epididymis and seminal vesicle. Blood and semen samples were collected on days -2, -1, 1, 3, 5, 7, 14 and weekly thereafter, up to postinfection day (PID 84. The inoculated steers (GI and GII presented hyperthermia from PID 3 to 16. Antibodies against T. gondii were detected through the indirect fluorescence antibody test (IFAT on PID 5 (1:16 in both inoculated groups (oocysts and tachyzoites, reaching peaks of 1:4096 on PID 7. Parasitemia outbursts occurred in all infected bovines, principally from PID 7 to 28, independent of the strain and inoculate used. Bioassays revealed the presence of parasites in semen samples of animals infected with oocysts (GI and tachyzoites (GII on several experimental days between PID 7 and 84. Tissue parasitism by T. gondii was diagnosed by bioassay and the PCR technique in several organ and tissue fragments. These findings suggest the possibility of sexual transmission of T. gondii in the bovine species.Dezoito bovinos foram inoculados com Toxoplasma gondii e distribuídos aleatoriamente em três grupos de seis bovinos cada: GI (2,5x10(5 oocistos da cepa "P", GII (5,0x10(6 taquizoítos da cepa "RH" e GIII (controle. Exames clínicos, sorológicos e parasitêmicos foram realizados. Pesquisas do parasito, por meio da bioprova e pela técnica de Reação em Cadeia pela Polimerase (PCR, foram realizadas no sêmen e em fragmentos de musculatura esquelética, linfonodos, cérebro, retina, baço, fígado, pulmão, testículo, epidídimo e vesícula seminal. Amostras de sangue e sêmen foram

  19. Diagnosis of pulmonary infection with Toxoplasma gondii in immunocompromised HIV-positive patients by real-time PCR

    DEFF Research Database (Denmark)

    Petersen, E.; Edvinsson, B.; Lundgren, Bettina

    2006-01-01

    . In positive samples, the genotype of the parasite was determined by sequence analysis of the GRA6 gene. Positive results were achieved for 2% (7/332) of the samples tested. Genotyping was possible in two samples and revealed GRA6 type II T. gondii. PCR for detecting T. gondii in BAL samples should...... be performed in all immunosuppressed HIV-positive patients with symptoms of a systemic infection of unknown etiology. Trimethoprim-sulfamethoxazole prophylaxis does not exclude concomitant infection with T. gondii....

  20. Occurrence of infection by Toxoplasma gondii in slaughtered swine in the northwestern region of Paraná, Brazil

    Directory of Open Access Journals (Sweden)

    Hannah Lia Ettiene Peruch Lemos dos Santos

    2015-07-01

    Full Text Available This study aimed to determine the prevalence of Toxoplasma gondii in swine slaughtered in Iporã, Northwest Paraná state. Blood samples were obtained from 500 finishing swine. All animals were raised under intensive farming; the blood samples were analyzed using the immunofluorescence antibody test (IFAT. Animals with titers of ?64 were considered positive for T. gondii infection. Of the tested samples, 63 (12.6% were positive by IFAT; 58 (92.06% of these showed titers of 64 (4.7%, with titers 256 and two (3.1% titers of 1024. These pigs might be considered a source of T. gondii infection for humans

  1. A new set of primers for the detection of Toxoplasma gondii in amniotic fluid using polymerase chain reaction.

    Science.gov (United States)

    Pelloux, H; Weiss, J; Simon, J; Muet, F; Fricker-Hidalgo, H; Goullier-Fleuret, A; Ambroise-Thomas, P

    1996-04-15

    A new PCR system including a pair of primers, a probe and an internal control were designed from the B1 gene of Toxoplasma gondii. The system described allowed the detection of less than 10 tachyzoites of the RH strain of T. gondii. Among 21 amniotic fluid samples, this system diagnosed the cases of congenital toxoplasmosis which were simultaneously diagnosed using mice inoculation, in vitro culture, and serology from both amniotic fluid and fetal blood. These results show that these new primers allow for a highly sensitive detection of T. gondii DNA.

  2. Cyclosporine A suppresses immunoglobulin G biosynthesis via inhibition of cyclophilin B in murine hybridomas and B cells.

    Science.gov (United States)

    Lee, Jisun; Choi, Tae Gyu; Ha, Joohun; Kim, Sung Soo

    2012-01-01

    Immunoglubulin G (IgG) is a major isotype of antibody, which is predominantly involved in immune response. The complete tetramer is needed to fold and assemble in endoplasmic reticulum (ER) prior to secretion from cells. Protein quality control guided by ER chaperons is most essential for full biological activity. Cyclophilin B (CypB) was initially identified as a high-affinity binding protein for the immunosuppressive drug Cyclosporine A (CsA). CsA suppresses organ rejection by halting productions of pro-inflammatory molecules in T cell and abolishes the enzymatic property of CypB that accelerates the folding of proteins by catalysing the isomerization of peptidyl-proline bonds in ER. Here, we reported that CsA significantly inhibited IgG biosynthesis at posttranslational level in antibody secreting cells. Moreover, CsA stimulated the extracellular secretion of CypB and induced ROS generation, leading to expressions of ER stress markers. In addition, the absence of intracellular CypB impaired the formation of ER multiprotein complex, which is most important for resisting ER stress. Interestingly, CsA interrupted IgG folding via occupying the PPIase domain of CypB in ER. Eventually, unfolded IgG is degraded via Herp-dependent ERAD pathway. Furthermore, IgG biosynthesis was really abrogated by inhibition of CypB in primary B cells. We established for the first time the immunosuppressive effect of CsA on B cells. Conclusively, the combined results of the current study suggest that CypB is a pivotal molecule for IgG biosynthesis in ER quality control. Copyright © 2011 Elsevier B.V. All rights reserved.

  3. Vital role for cyclophilin B (CypB) in asexual development, dimorphic transition and virulence of Beauveria bassiana.

    Science.gov (United States)

    Chu, Zhen-Jian; Sun, Huan-Huan; Ying, Sheng-Hua; Feng, Ming-Guang

    2017-08-01

    Cyclophilin B (CypB) was previously revealed as one of many putative secretory proteins in the transcriptome of Beauveria bassiana infection to a lepidopteran pest. Here we show a main localization of CypB in hyphal cell walls and septa and its essential role in the in vitro and in vivo asexual cycles of the fungal insect pathogen. Deletion of cypB reduced colony growth by 16-42% on two rich media and 30 scant media with different carbon or nitrogen sources. The deletion mutant suffered a delayed conidiation on a standard medium and a final 47% reduction in conidial yield, accompanied with drastic transcript depression of several key genes required for conidiation and conidial maturation. The mutant conidia required 10h longer to germinate 50% at optimal 25°C than wild-type conidia. Intriguingly, cultivation of the mutant conidia in a trehalose-peptone broth mimic to insect hemolymph resulted in 83% reduction in blastospore yield but only slight decrease in biomass level, indicating severe defects in transition of hyphae to blastospores. LT 50 for the deletion mutant against Galleria mellonella larvae through normal cuticle infection was prolonged to 7.4d from a wild-type estimate of 4.7d. During colony growth, additionally, the deletion mutant displayed hypersensitivity to Congo red, menadione, H 2 O 2 and heat shock but increased tolerance to cyclosporine A and rapamycin. All of changes were restored by targeted gene complementation. Altogether, CypB takes part in sustaining normal growth, aerial conidiation, conidial germination, dimorphic transition, stress tolerance and pathogenicity in B. bassiana. Copyright © 2017 Elsevier Inc. All rights reserved.

  4. Synthesis of Heparan Sulfate with Cyclophilin B-binding Properties Is Determined by Cell Type-specific Expression of Sulfotransferases*

    Science.gov (United States)

    Deligny, Audrey; Denys, Agnès; Marcant, Adeline; Melchior, Aurélie; Mazurier, Joël; van Kuppevelt, Toin H.; Allain, Fabrice

    2010-01-01

    Cyclophilin B (CyPB) induces migration and adhesion of T lymphocytes via a mechanism that requires interaction with 3-O-sulfated heparan sulfate (HS). HS biosynthesis is a complex process with many sulfotransferases involved. N-Deacetylases/N-sulfotransferases are responsible for N-sulfation, which is essential for subsequent modification steps, whereas 3-O-sulfotransferases (3-OSTs) catalyze the least abundant modification. These enzymes are represented by several isoforms, which differ in term of distribution pattern, suggesting their involvement in making tissue-specific HS. To elucidate how the specificity of CyPB binding is determined, we explored the relationships between the expression of these sulfotransferases and the generation of HS motifs with CyPB-binding properties. We demonstrated that high N-sulfate density and the presence of 2-O- and 3-O-sulfates determine binding of CyPB, as evidenced by competitive experiments with heparin derivatives, soluble HS, and anti-HS antibodies. We then showed that target cells, i.e. CD4+ lymphocyte subsets, monocytes/macrophages, and related cell lines, specifically expressed high levels of NDST2 and 3-OST3 isoforms. Silencing the expression of NDST1, NDST2, 2-OST, and 3-OST3 by RNA interference efficiently decreased binding and activity of CyPB, thus confirming their involvement in the biosynthesis of binding sequences for CyPB. Moreover, we demonstrated that NDST1 was able to partially sulfate exogenous substrate in the absence of NDST2 but not vice versa, suggesting that both isoenzymes do not have redundant activities but do have rather complementary activities in making N-sulfated sequences with CyPB-binding properties. Altogether, these results suggest a regulatory mechanism in which cell type-specific expression of certain HS sulfotransferases determines the specific binding of CyPB to target cells. PMID:19940140

  5. Synthesis of heparan sulfate with cyclophilin B-binding properties is determined by cell type-specific expression of sulfotransferases.

    Science.gov (United States)

    Deligny, Audrey; Denys, Agnès; Marcant, Adeline; Melchior, Aurélie; Mazurier, Joël; van Kuppevelt, Toin H; Allain, Fabrice

    2010-01-15

    Cyclophilin B (CyPB) induces migration and adhesion of T lymphocytes via a mechanism that requires interaction with 3-O-sulfated heparan sulfate (HS). HS biosynthesis is a complex process with many sulfotransferases involved. N-Deacetylases/N-sulfotransferases are responsible for N-sulfation, which is essential for subsequent modification steps, whereas 3-O-sulfotransferases (3-OSTs) catalyze the least abundant modification. These enzymes are represented by several isoforms, which differ in term of distribution pattern, suggesting their involvement in making tissue-specific HS. To elucidate how the specificity of CyPB binding is determined, we explored the relationships between the expression of these sulfotransferases and the generation of HS motifs with CyPB-binding properties. We demonstrated that high N-sulfate density and the presence of 2-O- and 3-O-sulfates determine binding of CyPB, as evidenced by competitive experiments with heparin derivatives, soluble HS, and anti-HS antibodies. We then showed that target cells, i.e. CD4+ lymphocyte subsets, monocytes/macrophages, and related cell lines, specifically expressed high levels of NDST2 and 3-OST3 isoforms. Silencing the expression of NDST1, NDST2, 2-OST, and 3-OST3 by RNA interference efficiently decreased binding and activity of CyPB, thus confirming their involvement in the biosynthesis of binding sequences for CyPB. Moreover, we demonstrated that NDST1 was able to partially sulfate exogenous substrate in the absence of NDST2 but not vice versa, suggesting that both isoenzymes do not have redundant activities but do have rather complementary activities in making N-sulfated sequences with CyPB-binding properties. Altogether, these results suggest a regulatory mechanism in which cell type-specific expression of certain HS sulfotransferases determines the specific binding of CyPB to target cells.

  6. Expression of cyclophilin B is associated with malignant progression and regulation of genes implicated in the pathogenesis of breast cancer.

    Science.gov (United States)

    Fang, Feng; Flegler, Ayanna J; Du, Pan; Lin, Simon; Clevenger, Charles V

    2009-01-01

    Cyclophilin B (CypB) is a 21-kDa protein with peptidyl-prolyl cis-trans isomerase activity that functions as a transcriptional inducer for Stat5 and as a ligand for CD147. To better understand the global function of CypB in breast cancer, T47D cells with a small interfering RNA-mediated knockdown of CypB were generated. Subsequent expression profiling analysis showed that 663 transcripts were regulated by CypB knockdown, and that many of these gene products contributed to cell proliferation, cell motility, and tumorigenesis. Real-time PCR confirmed that STMN3, S100A4, S100A6, c-Myb, estrogen receptor alpha, growth hormone receptor, and progesterone receptor were all down-regulated in si-CypB cells. A linkage analysis of these array data to protein networks resulted in the identification of 27 different protein networks that were impacted by CypB knockdown. Functional assays demonstrated that CypB knockdown also decreased cell growth, proliferation, and motility. Immunohistochemical and immunofluorescent analyses of a matched breast cancer progression tissue microarray that was labeled with an anti-CypB antibody demonstrated a highly significant increase in CypB protein levels as a function of breast cancer progression. Taken together, these results suggest that the enhanced expression of CypB in malignant breast epithelium may contribute to the pathogenesis of this disease through its regulation of the expression of hormone receptors and gene products that are involved in cell proliferation and motility.

  7. UNC93B1 mediates host resistance to infection with Toxoplasma gondii.

    Directory of Open Access Journals (Sweden)

    Mariane B Melo

    2010-08-01

    Full Text Available UNC93B1 associates with Toll-Like Receptor (TLR 3, TLR7 and TLR9, mediating their translocation from the endoplasmic reticulum to the endolysosome, hence allowing proper activation by nucleic acid ligands. We found that the triple deficient '3d' mice, which lack functional UNC93B1, are hyper-susceptible to infection with Toxoplasma gondii. We established that while mounting a normal systemic pro-inflammatory response, i.e. producing abundant MCP-1, IL-6, TNFα and IFNγ, the 3d mice were unable to control parasite replication. Nevertheless, infection of reciprocal bone marrow chimeras between wild-type and 3d mice with T. gondii demonstrated a primary role of hemopoietic cell lineages in the enhanced susceptibility of UNC93B1 mutant mice. The protective role mediated by UNC93B1 to T. gondii infection was associated with impaired IL-12 responses and delayed IFNγ by spleen cells. Notably, in macrophages infected with T. gondii, UNC93B1 accumulates on the parasitophorous vacuole. Furthermore, upon in vitro infection the rate of tachyzoite replication was enhanced in non-activated macrophages carrying mutant UNC93B1 as compared to wild type gene. Strikingly, the role of UNC93B1 on intracellular parasite growth appears to be independent of TLR function. Altogether, our results reveal a critical role for UNC93B1 on induction of IL-12/IFNγ production as well as autonomous control of Toxoplasma replication by macrophages.

  8. Canine distemper virus and Toxoplasma gondii co-infection in dogs with neurological signs

    Directory of Open Access Journals (Sweden)

    D.M. Aguiar

    2012-02-01

    Full Text Available O presente estudo relata a ocorrência de co-infecção entre o vírus da cinomose canina (CDV e Toxoplama gondii em cães com sinais neurológicos. Amostras de soro e tecido nervoso (pos-mortem de 21 cães, suspeitos de cinomose canina foram analisadas pela Reação de Imunofluorecência indireta (RIFI para pesquisa de anticorpos contra T. gondii e N. caninum e por RT-PCR para CDV. Dezessete (80,9% cães foram positivos para o CDV pela RT-PCR e 8 (38,1% foram positivos para anticorpos contra T. gondii. Sete cães (41,1% apresentaram-se positivos para ambos agentes, caracterizando processo de co-infecção. Somente 1 (4,7% cão foi soropositivo para N. caninum (RIFI=100, entretanto este mesmo animal foi positivo para T. gondii (RIFI=4096 e para CDV (RT-PCR.

  9. Frequency of exposure of endangered Caspian seals to Canine distemper virus, Leptospira interrogans, and Toxoplasma gondii.

    Directory of Open Access Journals (Sweden)

    Somayeh Namroodi

    Full Text Available Canine distemper virus (CDV, Leptospira interrogans, and Toxoplasma gondii are potentially lethal pathogens associated with decline in marine mammal populations. The Caspian Sea is home for the endangered Caspian seal (Pusa caspica. In the late 1990s and early 2000s, CDV caused a series of mortality events involving at least several thousand Caspian seals. To assess current infection status in Caspian seals, we surveyed for antibodies to three pathogens with potential to cause mortality in marine mammals. During 2015-2017, we tested serum samples from 36, apparently healthy, Caspian seals, accidentally caught in fishing nets in the Caspian Sea off Northern Iran, for antibodies to CDV, L. interrogans, and T. gondii, by virus neutralization, microscopic agglutination, and modified agglutination, respectively. Twelve (33%, 6 (17%, and 30 (83% samples were positive for CDV, L. interrogans and T. gondii antibodies, respectively. The highest titers of CDV, L. interrogans, and T. gondii antibodies were 16, 400, and 50, respectively. Frequencies of antibody to these pathogens were higher in seals >1 year old compared to seals <1 year old. Two serovars of L. interrogans (Pomona and Canicola were detected. Our results suggest a need for additional studies to clarify the impact of these pathogens on Caspian seal population decline and the improvement of management programs, including systematic screening to detect and protect the remaining population from disease outbreaks.

  10. Cytomegalovirus and Toxoplasma gondii seroprevalence in a Brazilian liver transplant waiting

    Directory of Open Access Journals (Sweden)

    R. A. M. B. Almeida

    2007-01-01

    Full Text Available Cytomegalovirus (CMV disease is a major cause of morbidity and mortality in solid organ transplantation. Disseminated toxoplasmosis after liver transplantation is a rare but fatal event. Serologic screening of the donor and the recipient is essential to prophylactic management, early diagnosis and therapeutic strategies to minimize the consequences of these infections. The aim of the present study was to determine the seroprevalence of CMV and Toxoplasma gondii (TG in a Brazilian liver transplant waiting list (LTWL. Serological data were collected from 44 candidates on the LTWL between May 2003 and November 2004. Serological investigation of antibodies IgM and IgG against CMV (anti-CMV and TG (anti-T. gondii was performed using fluorometry commercial kits. IgG anti-CMV was positive in 37 patients (94.9% out of 39 available results. There were not IgM anti-CMV positive results. Out of 36 analyzed patients, 22 (61.1% presented positive IgG anti-T. gondii and none had positive IgM anti-T. gondii. The high CMV seroprevalence among our LTWL reinforces the need for appropriate protocols to avoid related complications, like reactivation and superinfection by CMV. Environmental and drug prophylactic strategies against primary infection and reactivation, as well as early diagnosis and treatment of toxoplasmosis complications, are essential for the good outcome of transplant patients.

  11. Uptake and transmission of Toxoplasma gondii oocysts by migratory filter-feeding fish

    Science.gov (United States)

    Toxoplasma gondii is a ubiquitous parasitic protozoan known to cause disease and death in warm-blooded animals. Bottlenose dolphins, walruses, sea otters, and other marine animals worldwide have died from toxoplasmosis, but the source of this parasite in the marine environment h...

  12. Low Seroprevalence of Leishmania infantum and Toxoplasma gondii in the Horse Population in Israel

    NARCIS (Netherlands)

    Aharonson-Raz, Karin; Baneth, Gad; Lopes, Ana Patrícia; Brancal, Hugo; Schallig, Henk; Cardoso, Luís; Steinman, Amir

    2015-01-01

    A cross-sectional investigation was done on the seroprevalence of Leishmania infantum and Toxoplasma gondii infection among apparently healthy horses in Israel. This survey included 383 horses distributed in 22 farms throughout Israel during the years 2011-2013. Serum samples were tested for the

  13. Infectivity of cysts of the ME-49 Toxoplasma gondii strain in bovine milk and homemade cheese

    Directory of Open Access Journals (Sweden)

    Hiramoto RM

    2001-01-01

    Full Text Available OBJECTIVE: Analyze the infectivity and storage resistance of cysts of the ME-49 strain of Toxoplasma gondii in artificially infected bovine milk and homemade fresh cheese. METHODS: Pasteurized bovine milk was infected with 10 cysts/ml of the ME-49 strain of T.gondii and inoculated in different groups of mice, immediately or after storage at 4ºC for 5, 10 and 20 days. Homemade fresh cheese was prepared with artificially infected milk, and also tested in groups of mice, using the same storage process. Infection was identified by the presence of cysts in the brain or serological testing in challenged mice after 5 weeks, confirmed by Western Blot and histology. RESULTS: The infectivity of cysts of the ME-49 strain of T.gondii was maintained in the milk even after storage for 20 days at refrigerator temperatures. Cysts were also able to survive the production process of homemade fresh cheese and storage for a period of 10 days in the same conditions. CONCLUSIONS: These data demonstrated that milk and dairy products could be an important source of T.gondii in human contamination, reinforcing the importance of milk pasteurization before any processing or ingestion.

  14. Toxoplasma gondii in stranded marine mammals from the North Sea and Eastern Atlantic Ocean

    NARCIS (Netherlands)

    Velde, van de Norbert; Devleesschauwer, Brecht; Leopold, Mardik; Begeman, Lineke; IJsseldijk, Lonneke; Hiemstra, Sjoukje; IJzer, Jooske; Brownlow, Andrew; Davison, Nicholas; Haelters, Jan

    2016-01-01

    The occurrence of the zoonotic protozoan parasite Toxoplasma gondii in marine mammals remains a poorly understood phenomenon. In this study, samples from 589 marine mammal species and 34 European otters (Lutra lutra), stranded on the coasts of Scotland, Belgium, France, The Netherlands and

  15. Serological Evidence of Toxoplasma gondii infection in five species of bats in China

    Science.gov (United States)

    Toxoplasma gondii is an obligate intracellular protozoan parasite which can infect almost all warm-blooded animals and humans with a worldwide distribution. Bats are reservoirs for an increasing number of emerging zoonotic viruses, such as henipaviruses and severe acute respiratory syndrome (SARS) c...

  16. PREVALENCE OF ANTIBODIES AGAINST TOXOPLASMA GONDII IN POLAR BEARS (URSUS MARITIMUS) FROM SVALBARD AND EAST GREENLAND

    Science.gov (United States)

    Serum samples from 419 polar bears (Ursus maritimus) from Svalbard and the Barents Sea (collected 1990 - 2000) and 108 polar bears from East Greenland (collected 1999 - 2004) were assayed for antibodies against Toxoplasma gondii using the modified agglutination test (MAT). Antibody prevalences were ...

  17. Isolation of Toxoplasma gondii in a pig from a local abattoir in ...

    African Journals Online (AJOL)

    Inoculation of pepsin digested muscle (mixed heart and diaphragm) from a slaughter pig into six Balb C mice resulted in the isolation of Toxoplasma gondii which proved to be pathogenic to mice. These infected mice were retarded in growth when compared to the two uninoculated control animals. Mouse No.3 and Mouse ...

  18. Frequency of Toxoplasma gondii in the retina in eye banks in Brazil.

    Science.gov (United States)

    Costa, Deise F; Nascimento, Heloisa; Sutili, Aline; Nobrega, Fernando A J; Fowler, Flavio; Nobrega, Mario Junqueira; Garrido, Cristina; de Oliveira Dias, Janaina; Adán, Consuelo B D; Rizzo, Luiz Vicente; Silveira, Claudio; Belfort, Rubens; Commodaro, Alessandra G

    2017-07-01

    Ocular toxoplasmosis is the main cause of posterior uveitis worldwide frequently leading to vision loss. In Brazil, the seroprevalence of Toxoplasma gondii infection ranges from 50 to 80% depending of the region studied. The frequency of toxoplasmic retinal scar may reach 18% of the adults in the South of Brazil. Our goal was to determine the frequency of T. gondii DNA in retinas from eye banks from different regions in Brazil. A total of 162 eyes were obtained from eye banks in Manaus (n = 60), Sao Paulo (n = 60), Chapeco (n = 26), and Joinville (n = 16). The retinas were macroscopically analyzed and collected for DNA extraction. Real-time PCR (qPCR) was performed using the T. gondii B1 marker. By qPCR, a higher frequency of T. gondii DNA in the retinas from the eye bank of Joinville (25%) was found when compared to Manaus (5%). The retinas from Sao Paulo and Chapeco were qPCR negative. Clinical examination determined the retina lesions to be compatible with toxoplasmosis in the following frequencies: Joinville (62.5%), Manaus (10%), Sao Paulo (6.7%), and Chapeco (15.4%).

  19. Seroprevalence of Toxoplasma gondii in American Black Bears ( Ursus americanus ) of the Central Appalachians, USA.

    Science.gov (United States)

    Cox, John J; Murphy, Sean M; Augustine, Ben C; Guthrie, Joseph M; Hast, John T; Maehr, Sutton C; McDermott, Joseph

    2017-07-01

    We assessed Toxoplasma gondii seroprevalence in 53 free-ranging American black bears ( Ursus americanus ) in the Central Appalachian Mountains, US. Seroprevalence was 62% with no difference between males and females or between juvenile and adult bears. Wildlife agencies should consider warnings in hunter education programs to reduce the chances for human infection from this source.

  20. Kennel dogs as sentinels of Leishmania infantum, Toxoplasma gondii and Neospora caninum in Majorca Island, Spain

    Science.gov (United States)

    Kennel dogs can serve as sentinels and/or reservoirs of diseases of veterinary and zoonotic interest because they have often roamed free and lived outdoors, being exposed to pathogens. We tested for evidence of infection with three protozoans, Leishmania infantum, Toxoplasma gondii and Neospora cani...

  1. Behavioural and biochemical profiles in rats infected with Toxoplasma gondii or lipopolysaccharide: animal models of schizophrenia

    Czech Academy of Sciences Publication Activity Database

    Tejkalová, H.; Kodym, P.; Kačer, P.; Klaschka, Jan; Horáček, J.

    2013-01-01

    Roč. 24, e-Suppl. 1 (2013), e22 ISSN 0955-8810. [Biennal meeting of the EBPS /15./. 06.09.2013-09.09.2013, La Rochelle] Institutional support: RVO:67985807 Keywords : animal models * schizophrenia * behaviour * LPS * Toxoplasma gondii Subject RIV: FH - Neurology

  2. Conjugated Linoleic Acid Stimulates Apoptosis in RH and Tehran Strains of Toxoplasma gondii, in Vitro.

    Directory of Open Access Journals (Sweden)

    Jebreil Shamseddin

    2015-06-01

    Full Text Available The aim of this study was to evaluate the effects of conjugated linoleic acid (CLA on apoptosis of tachyzoites of T. gondii, RH strain (type I and the cyst-forming Tehran strain (type II in vitro.Toxoplasma strains were injected into the peritoneal cavity of BALB/c mice. The Tehran strain forms cysts in the brain of mice. Bradyzoites within the cysts are reactivated to proliferative tachyzoites, by dexamethasone. Tachyzoites were aspirated from the peritoneum of infected mice, and the percentage of viable parasites was estimated with trypan blue staining. Tachyzoites were inoculated into HeLa cells cultivated in DMEM medium. Different concentrations of CLA were evaluated on T. gondii in HeLa cells by the tetrazolium (MTT colorimetric assay. Differentiation between apoptosis and cell death was determined by flow cytometry using Annexin V and propidium iodide (PI double staining. The statistical analysis performed by GraphPad Prism version 6.00.CLA induces apoptosis in virulent (RH and avirulent (Tehran strains of T. gondii. The results of MTT indicated that CLA could decrease the proliferation of tachyzoites of both strains in HeLa cells.Conjugated linoleic acid has anti-toxoplasmacidal activity on tachyzoites of T. gondii. Therefore, we recommended further studies on this component in order to achieve a new drug against the parasite.

  3. Association of Toxoplasma gondii infection with schizophrenia and its relationship with suicide attempts in these patients.

    Science.gov (United States)

    Ansari-Lari, Maryam; Farashbandi, Hassan; Mohammadi, Fahimeh

    2017-10-01

    To investigate the association between schizophrenia and Toxoplasma gondii, and to assess the association of infection with suicide attempts and age of onset of schizophrenia in these patients. Case-control study Fars Province, southern Iran. Cases were individuals with psychiatric diagnosis of schizophrenia as per Diagnostic and Statistical Manual of Mental Disorders (DSM-IV) criteria. Controls were healthy blood donors, frequency-matched with patients according to age and sex. For the detection of IgG antibodies, enzyme-linked immunosorbent assay (ELISA) was used. Data about demographic information in all subjects and duration of illness and history of suicide attempts in patients with schizophrenia were collected using a brief questionnaire and hospital records. Chi-square test and multivariable logistic regression were used for statistical analyses. Among 99 cases, 42 individuals (42%) were positive for T. gondii antibody, vs. 41 (27%) among 152 controls (OR = 2, 95% CI: 1.2-3.4, P = 0.012). We compared the suicide attempts in patients with schizophrenia based on their T. gondii serologic status. There was a lower rate of suicide attempts in seropositive male patients than seronegative ones (OR = 0.3, 95% CI: 0.1-0.97, P = 0.04). Age of onset of schizophrenia did not differ between T. gondii-infected and non-infected patients. These findings may have implications for schizophrenia and suicide prevention programmes. However, clearly further studies are required to confirm them. © 2017 John Wiley & Sons Ltd.

  4. Toxoplasma gondii influences aversive behaviors of female rats in an estrus cycle dependent manner.

    Science.gov (United States)

    Golcu, Doruk; Gebre, Rahiwa Z; Sapolsky, Robert M

    2014-08-01

    The protozoan Toxoplasma gondii (T. gondii) manipulates the behavior of its rodent intermediate host to facilitate its passage to its feline definitive host. This is accomplished by a reduction of the aversive response that rodents show towards cat odors, which likely increases the predation risk. Females on average show similar changes as males. However, behaviors that relate to aversion and attraction are usually strongly influenced by the estrus cycle. In this study, we replicated behavioral effects of T. gondii in female rats, as well as expanded it to two novel behavioral paradigms. We also characterized the role of the estrus cycle in the behavioral effects of T. gondii on female rats. Uninfected females preferred to spend more time in proximity to rabbit rather than bobcat urine, and in a dark chamber rather than a lit chamber. Infected females lost both of these preferences, and also spent more time investigating social novelty (foreign bedding in their environment). Taken together, these data suggest that infection makes females less risk averse and more exploratory. Furthermore, this effect was influenced by the estrus cycle. Uninfected rats preferred rabbit urine to bobcat urine throughout the cycle except at estrus and metestrus. In contrast, infected rats lost this preference at every stage of the cycle except estrus. Commensurate with the possibility that this was a hormone-dependent effect, infected rats had elevated levels of circulating progesterone, a known anxiolytic. Copyright © 2014 Elsevier Inc. All rights reserved.

  5. Prevalence of antibodies to Leishmania infantum and Toxoplasma gondii in horses from the north of Portugal

    Science.gov (United States)

    Background Leishmania infantum and Toxoplasma gondii are protozoa with zoonotic and economic importance. Prevalences of antibodies to these agents were assessed in 173 horses from the north of Portugal. Findings Antibodies to L. infantum were detected by the direct agglutination test (DAT); seven (...

  6. Isolation of viable Toxoplasma gondii, molecular characterization, and seroprevalence in elk (Cervus canadensis) in Pennsylvania, USA

    Science.gov (United States)

    Toxoplasmosis is a worldwide zoonosis. The ingestion of uncooked/undercooked meat and consumption of water contaminated with Toxoplasma gondii oocysts excreted by felids are the main modes of transmission of this parasite. Thousands of wild cervids are hunted or killed in traffic accidents yearly bu...

  7. Serological Survey and Factors Associated with Toxoplasma gondii Infection in Domestic Goats in Myanmar

    Science.gov (United States)

    Bawm, Saw; Maung, Wint Yi; Win, Myat Yee; Thu, May June; Chel, Hla Myet; Khaing, Tin Aye; Wai, Soe Soe; Htun, Lat Lat; Myaing, Tin Tin; Tiwananthagorn, Saruda; Igarashi, Makoto; Katakura, Ken

    2016-01-01

    Goat farming is important for the livelihood of millions of rural people because it contributes to food security and creation of assets. However, infection of goats with Toxoplasma gondii could be a source of parasite transmission to humans. The information on T. gondii infection of goat was not reported yet in Myanmar. A total of 119 goat serum samples were collected from three cities in the central region of Myanmar for T. gondii antibody survey. With the occurrence value obtained in this first study, a second one, more complete, with larger number (162) of animals and properties, was carried out and the risk factors and prevalence were determined. In both studies the samples were analyzed by the LAT. Of these, 32 (11.4%) samples were showed to be positive. The infection was associated with the presence of cats at the farm (odds ratio [OR] = 4.66, 95% confidential interval [CI] = 1.03–21.06), farming with different animal species (sheep, cattle, and pigs) (OR = 4.33, 95% CI = 1.57–11.94), and farming without good management practices (OR = 0.23, 95% CI = 0.06–0.83). This is the first T. gondii prevalence study in goats in the country. PMID:26904362

  8. Cats and Goat Whey Associated with Toxoplasma gondii Infection in Pigs

    NARCIS (Netherlands)

    Meerburg, B.G.; Riel, van J.W.; Cornelissen, J.B.W.J.; Kijlstra, A.; Mul, M.F.

    2006-01-01

    In organic livestock production systems, farm-management factors are thought to play an important role in the on-farm prevalence of Toxoplasma gondii. Serological results and the results of an HACCP analysis were combined to determine important risk factors for the prevalence of this protozoan

  9. Serological Survey and Factors Associated with Toxoplasma gondii Infection in Domestic Goats in Myanmar

    Directory of Open Access Journals (Sweden)

    Saw Bawm

    2016-01-01

    Full Text Available Goat farming is important for the livelihood of millions of rural people because it contributes to food security and creation of assets. However, infection of goats with Toxoplasma gondii could be a source of parasite transmission to humans. The information on T. gondii infection of goat was not reported yet in Myanmar. A total of 119 goat serum samples were collected from three cities in the central region of Myanmar for T. gondii antibody survey. With the occurrence value obtained in this first study, a second one, more complete, with larger number (162 of animals and properties, was carried out and the risk factors and prevalence were determined. In both studies the samples were analyzed by the LAT. Of these, 32 (11.4% samples were showed to be positive. The infection was associated with the presence of cats at the farm (odds ratio [OR] = 4.66, 95% confidential interval [CI] = 1.03–21.06, farming with different animal species (sheep, cattle, and pigs (OR = 4.33, 95% CI = 1.57–11.94, and farming without good management practices (OR = 0.23, 95% CI = 0.06–0.83. This is the first T. gondii prevalence study in goats in the country.

  10. Prevalence of antibodies to Leishmania infantum and Toxoplasma gondii in horses from the north of Portugal

    NARCIS (Netherlands)

    Lopes, Ana Patrícia; Sousa, Susana; Dubey, Jp; Ribeiro, Ana J.; Silvestre, Ricardo; Cotovio, Mário; Schallig, Henk Dfh; Cardoso, Luís; Cordeiro-da-Silva, Anabela

    2013-01-01

    Leishmania infantum and Toxoplasma gondii are protozoa with zoonotic and economic importance. Prevalences of antibodies to these agents were assessed in 173 horses from the north of Portugal. Antibodies to L. infantum were detected by the direct agglutination test (DAT); seven (4.0%) horses were

  11. The Evolution of Host Mitochondrial Association and its Impact on Toxoplasma gondii Infection

    Science.gov (United States)

    English, Elizabeth D.

    The association of intracellular pathogens with host mitochondria has been observed across taxa, from bacterial pathogens, such as Legionella pneumophila and Chlamydia trachomati, to the eukaryotic pathogen Toxoplasma gondii. However the functional impact of host mitochondrial association (HMA) remains difficult to assess in most of these species because in many cases the genes responsible for this phenomenon have not yet been identified. The recent discovery of the T. gondii gene responsible for HMA, Mitochondrial Association Factor 1 ( MAF1) has provided us with the tools to begin to understand the evolution and impact of HMA. Here we use multispecies sequence analysis to determine that the MAF1 locus is tandemly duplicated and diversified in both T. gondii and its nearest extant relative Hammondia hammondi, but not another close relative Neospora caninum. Using cross-species complementation we find that T. gondii and H. hammondi harbor copies of MAF1 able to mediate HMA, while N. caninum does not. We have begun mutational analysis using naturally occurring HMA+ and HMA- paralogs of MAF1 in order to determine the portions of MAF1 protein necessary for HMA. Additionally, we have identified the first in vivo phenotypes associated with HMA using multiple mouse models, for both acute and chronic infection. Taken together these data indicate that HMA likely evolved via neofunctionalization of a duplicated ancestral MAF1 gene, and that the neofunctionalized, HMA competent copy of MAF1 provides a selective advantage.

  12. Frequency of exposure of endangered Caspian seals to Canine distemper virus, Leptospira interrogans, and Toxoplasma gondii.

    Science.gov (United States)

    Namroodi, Somayeh; Shirazi, Amir S; Khaleghi, Seyyed Reza; N Mills, James; Kheirabady, Vahid

    2018-01-01

    Canine distemper virus (CDV), Leptospira interrogans, and Toxoplasma gondii are potentially lethal pathogens associated with decline in marine mammal populations. The Caspian Sea is home for the endangered Caspian seal (Pusa caspica). In the late 1990s and early 2000s, CDV caused a series of mortality events involving at least several thousand Caspian seals. To assess current infection status in Caspian seals, we surveyed for antibodies to three pathogens with potential to cause mortality in marine mammals. During 2015-2017, we tested serum samples from 36, apparently healthy, Caspian seals, accidentally caught in fishing nets in the Caspian Sea off Northern Iran, for antibodies to CDV, L. interrogans, and T. gondii, by virus neutralization, microscopic agglutination, and modified agglutination, respectively. Twelve (33%), 6 (17%), and 30 (83%) samples were positive for CDV, L. interrogans and T. gondii antibodies, respectively. The highest titers of CDV, L. interrogans, and T. gondii antibodies were 16, 400, and 50, respectively. Frequencies of antibody to these pathogens were higher in seals >1 year old compared to seals <1 year old. Two serovars of L. interrogans (Pomona and Canicola) were detected. Our results suggest a need for additional studies to clarify the impact of these pathogens on Caspian seal population decline and the improvement of management programs, including systematic screening to detect and protect the remaining population from disease outbreaks.

  13. Antibodies to Toxoplasma gondii and Leishmania spp. in domestic cats from Luanda, Angola

    Science.gov (United States)

    Toxoplasma gondii and Leishmania spp. are zoonotic agents of importance to public health, with domestic cats as potential reservoirs for both protozoal infections. The present study aimed at assessing for the first time the seroprevalence of these zoonotic parasites in a domestic feline population l...

  14. Macrophages facilitate the excystation and differentiation of Toxoplasma gondii sporozoites into tachyzoites following oocyst internalization

    Science.gov (United States)

    Toxoplasma gondii is a common parasite of humans and domestic animals, which is transmitted via oocysts in cat faeces or tissue cysts in contaminated meat. The oocyst and sporocyst walls are multilayered polymeric structures that protect the infective sporozoites from deleterious physical and chemic...

  15. An experimental Toxoplasma gondii dose response challenge model to study therapeutic or vaccine efficacy in cats

    NARCIS (Netherlands)

    Cornelissen, J.B.W.J.; Giessen, van der J.W.B.; Takumi, K.; Teunis, P.F.M.; Wisselink, H.J.

    2014-01-01

    High numbers of Toxoplasma gondii oocysts in the environment are a risk factor to humans. The environmental contamination might be reduced by vaccinating the definitive host, cats. An experimental challenge model is necessary to quantitatively assess the efficacy of a vaccine or drug treatment.

  16. Toxoplasma gondii infection induces suppression in a mouse model of allergic airway inflammation.

    Directory of Open Access Journals (Sweden)

    Ignacio M Fenoy

    Full Text Available Allergic asthma is an inflammatory disorder characterized by infiltration of the airway wall with inflammatory cells driven mostly by activation of Th2-lymphocytes, eosinophils and mast cells. There is a link between increased allergy and a reduction of some infections in Western countries. Epidemiological data also show that respiratory allergy is less frequent in people exposed to orofecal and foodborne microbes such as Toxoplasma gondii. We previously showed that both acute and chronic parasite T. gondii infection substantially blocked development of airway inflammation in adult BALB/c mice. Based on the high levels of IFN-γ along with the reduction of Th2 phenotype, we hypothesized that the protective effect might be related to the strong Th1 immune response elicited against the parasite. However, other mechanisms could also be implicated. The possibility that regulatory T cells inhibit allergic diseases has received growing support from both animal and human studies. Here we investigated the cellular mechanisms involved in T. gondii induced protection against allergy. Our results show for the first time that thoracic lymph node cells from mice sensitized during chronic T. gondii infection have suppressor activity. Suppression was detected both in vitro, on allergen specific T cell proliferation and in vivo, on allergic lung inflammation after adoptive transference from infected/sensitized mice to previously sensitized animals. This ability was found to be contact-independent and correlated with high levels of TGF-β and CD4(+FoxP3(+ cells.

  17. Seropositivity of Toxoplasma gondii and Toxocara spp. in Children with Cryptogenic Epilepsy, Benha, Egypt.

    Science.gov (United States)

    Eraky, Maysa Ahmad; Abdel-Hady, Soha; Abdallah, Karim Fetouh

    2016-06-01

    The present study aimed to investigate the possible association of Toxoplasma gondii and Toxocara spp. infections with cryptogenic epilepsy in children. The study was carried out between June 2014 and March 2015. Total 90 children (40 with cryptogenic epilepsy, 30 with non-cryptogenic epilepsy, and 20 healthy control children) were evaluated to determine the anti-Toxocara and anti-T. gondii IgG seropositivity using ELISA kits. Epileptic cases were selected from those attending the pediatrics outpatient clinic of Benha University Hospital, Pediatrics Neurology Unit, and from Benha Specialized Hospital of children. The results showed that the level of anti-T. gondii IgG seropositivity was significantly higher among children with cryptogenic epilepsy (20%) than among children with non-cryptogenic children (0%). In healthy controls (10%), there was no association between toxocariasis seropositivity and cryptogenic epilepsy (only 5.7%; 4 out of 70 cases) among cases and 10% (2 out of 20) among controls. Among toxocariasis IgG positive cases, 3 (7.5%) were cryptogenic, and only 1 (3.3%) was non-cryptogenic. These statistically significant results support the association between T. gondii infection and cryptogenic epilepsy while deny this association with toxocariasis.

  18. Interaction and cystogenesis of Toxoplasma gondii within skeletal muscle cells in vitro

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    Erick Vaz Guimarães

    2009-03-01

    Full Text Available Infection by the protozoan parasite Toxoplasma gondii is widely prevalent in humans and animals. To prevent human infection, all meat should be well cooked before consumption, since the parasite is present in skeletal muscle. In this context, the use of skeletal muscle cells (SkMCs as a cellular model opens up new approaches to investigate T. gondii-host cell interactions. Immunofluorescent detection of proteins that are stage-specific for bradyzoites indicated that complete cystogenesis of T. gondii in in vitro cultures of SkMCs occurs after 96 h of infection. Ultrastructural analysis showed that, after 48 h of interaction, there were alterations on the parasitophorous vacuole membrane, including greater thickness and increased electron density at the inner face of the membrane. The present study demonstrates the potential use of primary cultures of SkMCs to evaluate different molecular aspects of T. gondii invasion and cystogenesis and presents a promising in vitro model for the screening of drug activities toward tissue cysts and bradyzoites.

  19. Antibodies to Toxoplasma gondii and Leishmania spp. in domestic cats from Luanda, Angola

    NARCIS (Netherlands)

    Lopes, Ana Patrícia; Oliveira, Ana Cristina; Granada, Sara; Rodrigues, Filipa T.; Papadopoulos, Elias; Schallig, Henk; Dubey, Jitender P.; Cardoso, Luís

    2017-01-01

    Toxoplasma gondii and Leishmania spp. are zoonotic protozoa of importance to animal and public health. The present study aimed to assess for the first time the seroprevalence of these zoonotic parasites in a domestic feline population living in Luanda, Angola. One hundred and two cats were sampled

  20. [Awareness and infection of Toxoplasma gondii in married childbearing women in Chengde Region].

    Science.gov (United States)

    Li, Xue-jing; Xu, Tao; Song, Ren-hao

    2014-08-01

    To understand Toxoplasma gondii infection and awareness condition of married childbearing women in Chengde Region, so as to provide the evidence for the establishment of control measures. Totally 733 married childbearing women who took physical examination in Chengde Hospital of Traditional Chinese Medicine from July to December in 2013 were investigated by questionnaire to understand the awareness condition on T. gondii infection, then 490 women among them from 3 counties and 2 districts were randomly chosen to detect the Toxoplasma antibodies by ELISA. A total of 733 questionnaires were returned, and 126 women knew related knowledge about T. gondii infection, and the awareness rate was 17.19%( 126/733). Sixty-three women were determined as infected cases, and the infection rate was 12.86%( 63/490). The infection rates of the women who with higher educational level, working as medical staff and living in urban were lower, and the awareness rates of them were higher. The infection rate of T. gondii among the married childbearing women in Chengde Region is high, and the awareness rate of them is low. In order to decrease the infection rate as well as to increase the awareness rate of the population, the health education should be strengthened.

  1. Presence of Toxoplasma gondii in Drinking Water from an Endemic Region in Southern Mexico.

    Science.gov (United States)

    Hernandez-Cortazar, Ivonne B; Acosta-Viana, Karla Y; Guzman-Marin, Eugenia; Ortega-Pacheco, Antonio; Segura-Correa, Jose C; Jimenez-Coello, Matilde

    2017-05-01

    Toxoplasmosis can be acquired through the ingestion of contaminated drinking water with oocysts of Toxoplasma gondii, highly resistant to the routinely disinfection processes; based on chlorination commonly used in the water supply industry. The aim of this study was to determine the presence of T. gondii DNA in samples of public drinking water from an endemic region of southern Mexico. In total 74 samples of water (5 L each) were collected from the three well fields (I, II, and III) and 71 independent wells, distributing public drinking water to the city of Merida Yucatan, after passing through the chlorination process. Water samples were filtered and concentrated by a sucrose solution, then DNA was extracted and evaluated through a nested-PCR (nPCR) specific for T. gondii. Positive samples were detected in 5.4% (4/74) of the water samples. This is the first report of the presence of T. gondii DNA in public drinking water from a large city in southern Mexico, where their consumption without any postpurification treatment could pose a risk for acquiring the infection in the urban population.

  2. Serology and genetics of Toxoplasma gondii in endangered Hawaiian (Nene) geese (Branta sandvicensis)

    Science.gov (United States)

    Toxoplasma gondii is parasite transmitted by feral cats that has historically caused mortality in native Hawaiian birds. A recent study revealed that this parasite accounts for ca. 4% of causes of mortality in native Hawaiian geese (nene-Branta sandvicensis). To know how widespread exposure to the...

  3. Targeted Delivery of Toxoplasma gondii Antigens to Dendritic Cells Promote Immunogenicity and Protective Efficiency against Toxoplasmosis

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    Zineb Lakhrif

    2018-02-01

    Full Text Available Toxoplasmosis is a major public health problem and the development of a human vaccine is of high priority. Efficient vaccination against Toxoplasma gondii requires both a mucosal and systemic Th1 immune response. Moreover, dendritic cells play a critical role in orchestrating the innate immune functions and driving specific adaptive immunity to T. gondii. In this study, we explore an original vaccination strategy that combines administration via mucosal and systemic routes of fusion proteins able to target the major T. gondii surface antigen SAG1 to DCs using an antibody fragment single-chain fragment variable (scFv directed against DEC205 endocytic receptor. Our results show that SAG1 targeting to DCs by scFv via intranasal and subcutaneous administration improved protection against chronic T. gondii infection. A marked reduction in brain parasite burden is observed when compared with the intranasal or the subcutaneous route alone. DC targeting improved both local and systemic humoral and cellular immune responses and potentiated more specifically the Th1 response profile by more efficient production of IFN-γ, interleukin-2, IgG2a, and nasal IgA. This study provides evidence of the potential of DC targeting for the development of new vaccines against a range of Apicomplexa parasites.

  4. Toxoplasma gondii antibodies in domiciled cats from rio branco Municipality, Acre State, Brazil

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    Soraia Figueiredo de Souza

    2015-12-01

    Full Text Available Blood samples were collected from 89 cats to assess the prevalence of IgG antibodies against Toxoplasma gondii by indirect immunofluorescence (IIF and the possible risk factors associated with feline Toxoplasma gondii infection. An epidemiological questionnaire was developed and implemented for owners of domestic cats domiciled in Rio Branco, Acre. The results were statistically evaluated with the odds ratio and chi-square tests, considering the significance level of 5%. Of 89 animals’ samples, 22 had antibodies against Toxoplasma gondii. Among the 22 reactive animals, 15 (68.19% were female, 15 (68.19% were less than one year old and 20 (90% were cross breed. Concerning risk factors, there was no difference (p > 0.05 between the variables evaluated by the chi-square test. Moreover, 16 (72% cats were fed a mixed diet, 20 (90% of the cats had hunting habits, 18 (81% had contact with animals of another species, 11 (50% had access to the street, and 22 (95% lived in homes that had areas of grass or dirt. In conclusion, the prevalence of anti-Toxoplasma gondii in domestic cats was 22.7%, and there were no significant risk factors for feline toxoplasmosis in the municipality of Rio Branco, Acre.

  5. Lacrimal secretory IgA in active posterior uveitis induced by Toxoplasma gondii

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    Maria Isabel Lynch

    2004-12-01

    Full Text Available It is quite difficult to diagnose active toxoplasmosis in patients with ocular toxoplasmosis. Active posterior uveitis presumably due to Toxoplasma gondii infection (APUPT is seldom produced during a prime-infection; hence most patients do not show high IgM antibodies. High levels of IgA have been described in active toxoplasmosis. The purpose of this study was to investigate possible association between APUPT and the specific anti-parasite sIgA in tears. The study was carried out as case-control. Tears of 25 clinically confirmed APUPT patients and 50 healthy control subjects were analyzed. All were IgG seropositive. Specific sIgA was determined by ELISA assay using T. gondii RH strain crude extract. Anti-T. gondii sIgA was found in 84% of the cases and in 22% of the control subjects. The intensity of the reaction was higher in APUPT cases (P = 0.007. There was strong association between APUPT patients and lacrimal sIgA (odds-ratio 18.61, P = 0.0001. ELISA test sensitivity was 84% and specificity 78% . Our data suggest that anti-T.gondii secretory IgA found in tears may become an important marker for active ocular toxoplasmosis.

  6. Lacrimal secretory IgA in active posterior uveitis induced by Toxoplasma gondii.

    Science.gov (United States)

    Lynch, Maria Isabel; Cordeiro, Francisco; Ferreira, Silvana; Ximenes, Ricardo; Oréfice, Fernando; Malagueño, Elizabeth

    2004-12-01

    It is quite difficult to diagnose active toxoplasmosis in patients with ocular toxoplasmosis. Active posterior uveitis presumably due to Toxoplasma gondii infection (APUPT) is seldom produced during a prime-infection; hence most patients do not show high IgM antibodies. High levels of IgA have been described in active toxoplasmosis. The purpose of this study was to investigate possible association between APUPT and the specific anti-parasite sIgA in tears. The study was carried out as case-control. Tears of 25 clinically confirmed APUPT patients and 50 healthy control subjects were analyzed. All were IgG seropositive. Specific sIgA was determined by ELISA assay using T. gondii RH strain crude extract. Anti-T. gondii sIgA was found in 84% of the cases and in 22% of the control subjects. The intensity of the reaction was higher in APUPT cases (P = 0.007). There was strong association between APUPT patients and lacrimal sIgA (odds-ratio 18.61, P = 0.0001). ELISA test sensitivity was 84% and specificity 78%. Our data suggest that anti-T.gondii secretory IgA found in tears may become an important marker for active ocular toxoplasmosis.

  7. Toxoplasma gondii infection induces dendritic retraction in basolateral amygdala accompanied by reduced corticosterone secretion

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    Rupshi Mitra

    2013-03-01

    Pathological anxiety is thought to reflect a maladaptive state characterized by exaggerated fear. Naturally occurring perturbations that reduce fear can be crucial in the search for new treatments. The protozoan parasite Toxoplasma gondii invades rat brain and removes the fear that rats have of cat odors, a change believed to be parasitic manipulation of host behavior aimed at increasing parasite transmission. It is likely that mechanisms employed by T. gondii can be used as a heuristic tool to understand possible means of fear reduction in clinical settings. Male Long-Evans rats were infected with T. gondii and compared with sham-infected animals 8 weeks after infection. The amount of circulating plasma corticosterone and dendritic arborization of basolateral amygdala principal neurons were quantified. Previous studies have shown that corticosterone, acting within the basolateral amygdala, enhances the fear response to environmental stimuli. Here we show that T. gondii infection causes a dendritic retraction in basolateral amygdala neurons. Such dendritic retraction is accompanied by lower amounts of circulating corticosterone, both at baseline and when induced by an aversive cat odor. The concerted effects of parasitism on two pivotal physiological nodes of the fear response provide an animal model relevant to interactions between stress hormones and amygdalar plasticity.

  8. Activation of CD147 with Cyclophilin A Induces the Expression of IFITM1 through ERK and PI3K in THP-1 Cells

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    Ju-Young Kim

    2010-01-01

    Full Text Available CD147, as a receptor for Cyclophilins, is a multifunctional transmembrane glycoprotein. In order to identify genes that are induced by activation of CD147, THP-1 cells were stimulated with Cyclophilin A and differentially expressed genes were detected using PCR-based analysis. Interferon-induced transmembrane 1 (IFITM1 was detected to be induced and it was confirmed by RT-PCR and Western blot analysis. CD147-induced expression of IFITM1 was blocked by inhibitors of ERK, PI3K, or NF-κB, but not by inhibitors of p38, JNK, or PKC. IFITM1 appears to mediate inflammatory activation of THP-1 cells since cross-linking of IFITM1 with specific monoclonal antibody against it induced the expression of proinflammatory mediators such as IL-8 and MMP-9. These data indicate that IFITM1 is one of the pro-inflammatory mediators that are induced by signaling initiated by the activation of CD147 in macrophages and activation of ERK, PI3K, and NF-κB is required for the expression of IFITM1.

  9. Involvement of the N-terminal part of cyclophilin B in the interaction with specific Jurkat T-cell binding sites.

    Science.gov (United States)

    Mariller, C; Haendler, B; Allain, F; Denys, A; Spik, G

    1996-07-15

    Cyclophilin B (CyPB) is secreted in biological fluids such as blood or milk and binds to a specific receptor present on the human lymphoblastic cell line Jurkat and on human peripheral blood lymphocytes. This study was intended to specify the areas of CyPB that are involved in the interaction with the receptor. A synthetic peptide corresponding to the first 24 N-terminal amino acid residues of CyPB was shown to specifically recognize the receptor. Moreover, modification of Arg18 of CyPB by p-hydroxyphenlglyoxal led to a dramatic loss of affinity for the receptor. However, when this residue was replaced by an alanine residue using site-directed mutagenesis, no modification of the binding properties was found, suggesting that Arg18 is not directly involved but is sufficiently close to the interaction site to interfere with the binding when modified. Competitive binding experiments using a chimaeric protein made up of the 24 N-terminal amino acid residues of CyPB fused to the cyclophilin A core sequence confirmed the involvement of this region of CyPB in receptor binding.

  10. First isolation and genotyping of Toxoplasma gondii from bats (Mammalia: Chiroptera).

    Science.gov (United States)

    Cabral, A D; Gama, A R; Sodré, M M; Savani, E S M M; Galvão-Dias, M A; Jordão, L R; Maeda, M M; Yai, L E O; Gennari, S M; Pena, H F J

    2013-03-31

    There are currently no reports on the isolation and molecular examination of Toxoplasma gondii from bats. Here, we report the isolation and genotypic characterisation of two T. gondii isolates from bats. A total of 369 bats from different municipalities in São Paulo state, southeastern Brazil, were captured and euthanised, and collected tissues (heart and pectoral muscle) were processed for each bat or in pools of two or three bats and bioassayed in mice (a total of 283 bioassays). Eleven PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) markers were used to genotype positive samples: SAG1, SAG2 (5'-3'SAG2 and alt. SAG2), SAG3, BTUB, GRA6, L358, c22-8, c29-2, PK1, CS3 and Apico. The parasite was isolated from two bats from São Paulo city: an insectivorous bat, the velvety free-tailed bat Molossus molossus, and a hematophagous bat, the common vampire bat Desmodus rotundus. Isolates were designated TgBatBr1 and TgBatBr2, respectively. The genotype of the isolate from M. molossus (TgBatBr1) has been previously described in an isolate from a capybara from São Paulo state, and the genotype from the D. rotundus isolate (TgBatBr2) has already been identified in isolates from cats, chickens, capybaras, sheep, a rodent and a common rabbit from different Brazilian states, suggesting that this may be a common T. gondii lineage circulating in some Brazilian regions. Isolation of T. gondii from a hematophagous species is striking. This study reveals that bats can share the same isolates that are found in domesticated and wild terrestrial animals. This is the first report of the isolation and genotyping of T. gondii in chiropterans. Copyright © 2012 Elsevier B.V. All rights reserved.

  11. Global iTRAQ-based proteomic profiling of Toxoplasma gondii oocysts during sporulation.

    Science.gov (United States)

    Zhou, Chun-Xue; Zhu, Xing-Quan; Elsheikha, Hany M; He, Shuai; Li, Qian; Zhou, Dong-Hui; Suo, Xun

    2016-10-04

    Toxoplasma gondii is a medically and economically important protozoan parasite. However, the molecular mechanisms of its sporulation remain largely unknown. Here, we applied iTRAQ coupled with 2D LC-MS/MS proteomic analysis to investigate the proteomic expression profile of T. gondii oocysts during sporulation. Of the 2095 non-redundant proteins identified, 587 were identified as differentially expressed proteins (DEPs). Based on Gene Ontology enrichment and KEGG pathway analyses the majority of these DEPs were found related to the metabolism of amino acids, carbon and energy. Protein interaction network analysis generated by STRING identified ATP-citrate lyase (ACL), GMP synthase, IMP dehydrogenase (IMPDH), poly (ADP-ribose) glycohydrolase (PARG), and bifunctional dihydrofolate reductase-thymidylate synthase (DHFR-TS) as the top five hubs. We also identified 25 parasite virulence factors that were expressed at relatively high levels in sporulated oocysts compared to non-sporulated oocysts, which might contribute to the infectivity of mature oocysts. Considering the importance of oocysts in the dissemination of toxoplasmosis these findings may help in the search of protein targets with a key role in infectiousness and ecological success of oocysts, creating new opportunities for the development of better means for disease prevention. The development of new preventative interventions against T. gondii infection relies on an improved understanding of the proteome and chemical pathways of this parasite. To identify proteins required for the development of environmentally resistant and infective T. gondii oocysts, we compared the proteome of non-sporulated (immature) oocysts with the proteome of sporulated (mature, infective) oocysts. iTRAQ 2D-LC-MS/MS analysis revealed proteomic changes that distinguish non-sporulated from sporulated oocysts. Many of the differentially expressed proteins were involved in metabolic pathways and 25 virulence factors were identified

  12. Qualitative Assessment for Toxoplasma gondii Exposure Risk Associated with Meat Products in the United States.

    Science.gov (United States)

    Guo, Miao; Buchanan, Robert L; Dubey, Jitender P; Hill, Dolores E; Lambertini, Elisabetta; Ying, Yuqing; Gamble, H Ray; Jones, Jeffrey L; Pradhan, Abani K

    2015-12-01

    Toxoplasma gondii is a global protozoan parasite capable of infecting most warm-blooded animals. Although healthy adult humans generally have no symptoms, severe illness does occur in certain groups, including congenitally infected fetuses and newborns, immunocompromised individuals including transplant patients. Epidemiological studies have demonstrated that consumption of raw or undercooked meat products is one of the major sources of infection with T. gondii. The goal of this study was to develop a framework to qualitatively estimate the exposure risk to T. gondii from various meat products consumed in the United States. Risk estimates of various meats were analyzed by a farm-to-retail qualitative assessment that included evaluation of farm, abattoir, storage and transportation, meat processing, packaging, and retail modules. It was found that exposure risks associated with meats from free-range chickens, nonconfinement-raised pigs, goats, and lamb are higher than those from confinement-raised pigs, cattle, and caged chickens. For fresh meat products, risk at the retail level was similar to that at the farm level unless meats had been frozen or moisture enhanced. Our results showed that meat processing, such as salting, freezing, commercial hot air drying, long fermentation times, hot smoking, and cooking, are able to reduce T. gondii levels in meat products. whereas nitrite and/or nitrate, spice, low pH, and cold storage have no effect on the viability of T. gondii tissue cysts. Raw-fermented sausage, cured raw meat, meat that is not hot-air dried, and fresh processed meat were associated with higher exposure risks compared with cooked meat and frozen meat. This study provides a reference for meat management control programs to determine critical control points and serves as the foundation for future quantitative risk assessments.

  13. Polyparasitism Is Associated with Increased Disease Severity in Toxoplasma gondii-Infected Marine Sentinel Species

    Science.gov (United States)

    Gibson, Amanda K.; Raverty, Stephen; Lambourn, Dyanna M.; Huggins, Jessica; Magargal, Spencer L.; Grigg, Michael E.

    2011-01-01

    In 1995, one of the largest outbreaks of human toxoplasmosis occurred in the Pacific Northwest region of North America. Genetic typing identified a novel Toxoplasma gondii strain linked to the outbreak, in which a wide spectrum of human disease was observed. For this globally-distributed, water-borne zoonosis, strain type is one variable influencing disease, but the inability of strain type to consistently explain variations in disease severity suggests that parasite genotype alone does not determine the outcome of infection. We investigated polyparasitism (infection with multiple parasite species) as a modulator of disease severity by examining the association of concomitant infection of T. gondii and the related parasite Sarcocystis neurona with protozoal disease in wild marine mammals from the Pacific Northwest. These hosts ostensibly serve as sentinels for the detection of terrestrial parasites implicated in water-borne epidemics of humans and wildlife in this endemic region. Marine mammals (151 stranded and 10 healthy individuals) sampled over 6 years were assessed for protozoal infection using multi-locus PCR-DNA sequencing directly from host tissues. Genetic analyses uncovered a high prevalence and diversity of protozoa, with 147/161 (91%) of our sampled population infected. From 2004 to 2009, the relative frequency of S. neurona infections increased dramatically, surpassing that of T. gondii. The majority of T. gondii infections were by genotypes bearing Type I lineage alleles, though strain genotype was not associated with disease severity. Significantly, polyparasitism with S. neurona and T. gondii was common (42%) and was associated with higher mortality and more severe protozoal encephalitis. Our finding of widespread polyparasitism among marine mammals indicates pervasive contamination of waterways by zoonotic agents. Furthermore, the significant association of concomitant infection with mortality and protozoal encephalitis identifies polyparasitism as

  14. Polyparasitism is associated with increased disease severity in Toxoplasma gondii-infected marine sentinel species.

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    Amanda K Gibson

    2011-05-01

    Full Text Available In 1995, one of the largest outbreaks of human toxoplasmosis occurred in the Pacific Northwest region of North America. Genetic typing identified a novel Toxoplasma gondii strain linked to the outbreak, in which a wide spectrum of human disease was observed. For this globally-distributed, water-borne zoonosis, strain type is one variable influencing disease, but the inability of strain type to consistently explain variations in disease severity suggests that parasite genotype alone does not determine the outcome of infection. We investigated polyparasitism (infection with multiple parasite species as a modulator of disease severity by examining the association of concomitant infection of T. gondii and the related parasite Sarcocystis neurona with protozoal disease in wild marine mammals from the Pacific Northwest. These hosts ostensibly serve as sentinels for the detection of terrestrial parasites implicated in water-borne epidemics of humans and wildlife in this endemic region. Marine mammals (151 stranded and 10 healthy individuals sampled over 6 years were assessed for protozoal infection using multi-locus PCR-DNA sequencing directly from host tissues. Genetic analyses uncovered a high prevalence and diversity of protozoa, with 147/161 (91% of our sampled population infected. From 2004 to 2009, the relative frequency of S. neurona infections increased dramatically, surpassing that of T. gondii. The majority of T. gondii infections were by genotypes bearing Type I lineage alleles, though strain genotype was not associated with disease severity. Significantly, polyparasitism with S. neurona and T. gondii was common (42% and was associated with higher mortality and more severe protozoal encephalitis. Our finding of widespread polyparasitism among marine mammals indicates pervasive contamination of waterways by zoonotic agents. Furthermore, the significant association of concomitant infection with mortality and protozoal encephalitis identifies

  15. Risk factors related to Toxoplasma gondii seroprevalence in indoor-housed Dutch dairy goats.

    Science.gov (United States)

    Deng, Huifang; Dam-Deisz, Cecile; Luttikholt, Saskia; Maas, Miriam; Nielen, Mirjam; Swart, Arno; Vellema, Piet; van der Giessen, Joke; Opsteegh, Marieke

    2016-02-01

    Toxoplasma gondii can cause disease in goats, but also has impact on human health through food-borne transmission. Our aims were to determine the seroprevalence of T. gondii infection in indoor-housed Dutch dairy goats and to identify the risk factors related to T. gondii seroprevalence. Fifty-two out of ninety approached farmers with indoor-kept goats (58%) participated by answering a standardized questionnaire and contributing 32 goat blood samples each. Serum samples were tested for T. gondii SAG1 antibodies by ELISA and results showed that the frequency distribution of the log10-transformed OD-values fitted well with a binary mixture of a shifted gamma and a shifted reflected gamma distribution. The overall animal seroprevalence was 13.3% (95% CI: 11.7–14.9%), and at least one seropositive animal was found on 61.5% (95% CI: 48.3–74.7%) of the farms. To evaluate potential risk factors on herd level, three modeling strategies (Poisson, negative binomial and zero-inflated) were compared. The negative binomial model fitted the data best with the number of cats (1–4 cats: IR: 2.6, 95% CI: 1.1–6.5; > = 5 cats:IR: 14.2, 95% CI: 3.9–51.1) and mean animal age (IR: 1.5, 95% CI: 1.1–2.1) related to herd positivity. In conclusion, the ELISA test was 100% sensitive and specific based on binary mixture analysis. T. gondii infection is prevalent in indoor housed Dutch dairy goats but at a lower overall animal level seroprevalence than outdoor farmed goats in other European countries, and cat exposure is an important risk factor.

  16. Seroprevalence of toxoplasma gondii infection among patients admitted to al-zahra hospital, isfahan, iran

    International Nuclear Information System (INIS)

    Mohaghegh, M.A.; Yazdani, H.; Hadipour, M.; Namdar, F.

    2015-01-01

    Toxoplasma gondii (T. gondii) infection is one of the most common parasitic infections among humans and other warm-blooded animals worldwide. The aim of this study was to evaluate toxoplasmosis status in patients admitted to Al-Zahra hospital, Isfahan, Iran. Methods: This cross-sectional study was conducted from October 2012 to January 2015. During this period, 716 patients referred to Al-Zahra hospital in Isfahan city, Iran, were studied to investigate the IgG and IgM antibodies against T. gondii using ELISA kit. The data were analysed by Chi-square and Fishers exact tests. In addition, the relation of data with age and sex were also examined. Results: Among 716 patients, 21 patients (2.9%) had positive IgM and 288 patients (40.2%) had positive IgG titer against T. gondii. Data analysis by Chi-square and Fishers exact tests revealed that there was no significant relationship between IgG titer and age (p>0.05). Additionally, there was no relationship between IgM titer and age (p>0.05). The data showed that there was no relationship between IgG and IgM antibody titer and sex (p>0.05). Conclusion: The prevalence of toxoplasmosis in Isfahan inhabitants seems fairly high but it can be concluded that the rate of seropositive patient is moderate comparing to other regions of country. Accordingly, the authors propose that all sensitive patients have to be tested for T. gondii antibody in order to prevent the consequences of disease. (author)

  17. Toxoplasma gondii infection: seroprevalence and associated risk factors among primary schoolchildren in Lagos City, Southern Nigeria

    Directory of Open Access Journals (Sweden)

    Vincent Pam Gyang

    2015-02-01

    Full Text Available INTRODUCTION: Toxoplasma gondii infection has been described as the most widespread zoonotic infection of humans and other animals. Information concerning T. gondii infection among schoolchildren is unavailable in Lagos City, Nigeria. METHODS: This cross-sectional study investigated the seroprevalence and risk factors associated with T. gondii infection among primary schoolchildren (PSC from a community located in the center of Lagos, southern Nigeria, from November 2013 to March 2014. A total of 382 PSC were screened for the presence of sera anti-T. gondii antibodies using a latex agglutination test (TOXO Test-MT, Tokyo, Japan. A cutoff titer of ≥ 1:32 was considered positive, while titers ≥ 1:1,024 indicated high responders. Questionnaires were also used to obtain data on possible risk factors from parents/guardians. RESULTS: The overall seroprevalence was 24% (91/382, and 83.5% (76/91 of seropositive PSC were classified as high responders. Among the risk factors tested, including contact with cats and soil, consumption of raw meat and vegetables, and drinking unboiled water, none showed statistical significance after multivariate adjustment. No associations were observed among age, gender, body mass index (BMI, and parents' occupation/educational level. CONCLUSIONS: The findings in this study show evidence of active infection, and hence, there is need for urgent preventive measures in this city. Further investigation is required to clarify the transmission routes. Policy makers also need to initiate prevention and control programs to protect pregnant women and immunocompromised patients in particular because they are more severely affected by T. gondii infection.

  18. Ciprofloxacin Derivatives Affect Parasite Cell Division and Increase the Survival of Mice Infected with Toxoplasma gondii.

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    Erica S Martins-Duarte

    Full Text Available Toxoplasmosis, caused by the protozoan Toxoplasma gondii, is a worldwide disease whose clinical manifestations include encephalitis and congenital malformations in newborns. Previously, we described the synthesis of new ethyl-ester derivatives of the antibiotic ciprofloxacin with ~40-fold increased activity against T. gondii in vitro, compared with the original compound. Cipro derivatives are expected to target the parasite's DNA gyrase complex in the apicoplast. The activity of these compounds in vivo, as well as their mode of action, remained thus far uncharacterized. Here, we examined the activity of the Cipro derivatives in vivo, in a model of acute murine toxoplasmosis. In addition, we investigated the cellular effects T. gondii tachyzoites in vitro, by immunofluorescence and transmission electron microscopy (TEM. When compared with Cipro treatment, 7-day treatments with Cipro derivatives increased mouse survival significantly, with 13-25% of mice surviving for up to 60 days post-infection (vs. complete lethality 10 days post-infection, with Cipro treatment. Light microscopy examination early (6 and 24h post-infection revealed that 6-h treatments with Cipro derivatives inhibited the initial event of parasite cell division inside host cells, in an irreversible manner. By TEM and immunofluorescence, the main cellular effects observed after treatment with Cipro derivatives and Cipro were cell scission inhibition--with the appearance of 'tethered' parasites--malformation of the inner membrane complex, and apicoplast enlargement and missegregation. Interestingly, tethered daughter cells resulting from Cipro derivatives, and also Cipro, treatment did not show MORN1 cap or centrocone localization. The biological activity of Cipro derivatives against C. parvum, an apicomplexan species that lacks the apicoplast, is, approximately, 50 fold lower than that in T. gondii tachyzoites, supporting that these compounds targets the apicoplast. Our results

  19. Antibody Prevalence and Isolation of Viable Toxoplasma gondii from Raptors in the Southeastern USA.

    Science.gov (United States)

    Love, David; Kwok, Oliver C; Verma, Shiv Kumar; Dubey, Jitender P; Bellah, Jamie

    2016-07-01

    Raptors are good indicators of the prevalence of Toxoplasma gondii in the environment because they prey on small mammals and birds. These prey species are a major source of infection in domestic cats ( Felis catus ), which shed the environmentally resistant oocysts. We assessed T. gondii infection in 281 opportunistically available raptors at a rehabilitation facility between 2012 and 2014. Antibodies to T. gondii were assayed by a modified agglutination test (cutoff 1:25) and found in serum of 22/71 Red-tailed Hawks ( Buteo jamaicensis ), 25/54 Barred Owls ( Strix varia ), 9/41 Red-shouldered Hawks ( Buteo lineatus ), 13/28 Great Horned Owls ( Bubo virginianus ), 6/20 Broad-winged Hawks ( Buteo platypterus ), 2/16 Eastern Screech Owls (Megascops asio), 12/13 Bald Eagles ( Haliaeetus leucocephalus ), 6/12 Cooper's Hawks ( Accipiter cooperii ), 1/8 Black Vultures ( Coragyps atratus ), and 1/1 Golden Eagle ( Aquila chrysaetos ). Antibodies were not detected in 5 Barn Owls ( Tyto alba ), 3 American Kestrels ( Falco sparverius ), 1 Mississippi Kite ( Ictinia mississippiensis ), and 1 Osprey ( Pandion haliaetus ). Viable T. gondii was isolated from the tissues of 1 antibody-positive Barred Owl and identified as a strain having type II alleles at all 10 loci tested, except one (ToxoDB polymerase chain reaction-restriction fragment length polymorphism genotype 3). Type II strain is the most common strain in the US. Results of this study indicate a high prevalence of T. gondii in some raptor species and the first reported genotyping from a Barred Owl.

  20. Toxoplasma gondii seroprevalence in pet cats in Norway and risk factors for seropositivity.

    Science.gov (United States)

    Sævik, Bente Kristin; Krontveit, Randi Ingebjørg; Eggen, Kristine P; Malmberg, Nina; Thoresen, Stein I; Prestrud, Kristin W

    2015-12-01

    The aims of the study were to estimate Toxoplasma gondii seroprevalence in pet cats in Norway and to evaluate risk factors for seropositivity. Additionally, serum biochemistry and haematological variables for T gondii seropositive and seronegative cats were compared. A convenience sample of surplus sera submitted to the Central Laboratory, Norwegian University of Life Sciences, was collected. The samples were from healthy cats and cats with a variety of diseases. Analyses for IgG antibodies to T gondii were performed with a commercial direct agglutination test, with 1:40 as the threshold value. For risk factor analysis a logistic regression model of the relationship between predictors and the outcome was applied. One hundred and ninety-six of 478 cats were seropositive for T gondii, and the estimated seroprevalence in the study sample was 41.0% (95% confidence interval 36.6-45.4). Compared with domestic cats, pedigree cats had reduced risk for Toxoplasma seropositivity (odds ratio [OR] 0.42). Males had increased risk (OR 1.63) compared with females. The effect of age was highly significant, and an increase in the cats' age across the interquartile range (IQR; 52-160 months/4-13 years of age) doubled the risk of Toxoplasma seropositivity (OR 2.11). The risk for Toxoplasma seropositivity among cats living in Oslo was significantly reduced (OR 0.51) when compared with the rest of Norway. Pet cats in Norway appear to be commonly exposed to T gondii. Signalment and geographical region influenced the odds of Toxoplasma seropositivity, whereas health status did not. © ISFM and AAFP 2015.

  1. Loop-mediated isothermal amplification (LAMP): early detection of Toxoplasma gondii infection in mice.

    Science.gov (United States)

    Kong, Qing-Ming; Lu, Shao-Hong; Tong, Qun-Bo; Lou, Di; Chen, Rui; Zheng, Bin; Kumagai, Takashi; Wen, Li-Yong; Ohta, Nobuo; Zhou, Xiao-Nong

    2012-01-03

    Toxoplasmosis is a widespread zoonotic parasitic disease that occurs in both animals and humans. Traditional molecular assays are often difficult to perform, especially for the early diagnosis of Toxoplasma gondii infections. Here, we established a novel loop-mediated isothermal amplification targeting the 529 bp repeat element (529 bp-LAMP) to detect T. gondii DNA in blood samples of experimental mice infected with tachyzoites of the RH strain. The assay was performed with Bst DNA polymerase at 65°C for 1 h. The detection limit of the 529 bp-LAMP assay was as low as 0.6 fg of T. gondii DNA. The sensitivity of this assay was 100 and 1000 fold higher than that of the LAMP targeting B1 gene (B1-LAMP) and nested PCR targeting 529 bp repeat element (529 bp-nested PCR), respectively. The specificity of the 529 bp-LAMP assay was determined using the DNA samples of Trypanosoma evansi, Plasmodium falciparum, Paragonimus westermani, Schistosoma japonicum, Fasciola hepatica and Angiostrongylus cantonensis. No cross-reactivity with the DNA of any parasites was found. The assay was able to detect T. gondii DNA in all mouse blood samples at one day post infection (dpi). We report the following findings: (i) The detection limit of the 529 bp-LAMP assay is 0.6 fg of T. gondii DNA; (ii) The assay does not involve any cross-reactivity with the DNA of other parasites; (iii) This is the first report on the application of the LAMP assay for early diagnosis of toxoplasmosis in blood samples from experimentally infected mice. Due to its simplicity, sensitivity and cost-effectiveness for common use, we suggest that this assay should be used as an early diagnostic tool for health control of toxoplasmosis.

  2. Reproductive implications of exposure to Toxoplasma gondii and Neospora caninum in western grey kangaroos (Macropus fuliginosus ocydromus).

    Science.gov (United States)

    Mayberry, Chris; Maloney, Shane K; Mitchell, Jeff; Mawson, Peter R; Bencini, Roberta

    2014-04-01

    Australian marsupials are thought to be particularly vulnerable to pathologic impacts of Toxoplasma gondii, and they may be similarly affected by Neospora caninum. Pathology due to either organism could be expressed as reduced female reproductive performance. We studied adult female western grey kangaroos (Macropus fuliginosus ocydromus) from suburban Perth, Western Australia, between May 2006 and October 2008. We used indirect fluorescent antibody tests to look for evidence of exposure to T. gondii and N. caninum in M. fuliginosus ocydromus and tested the association between their reproductive performance and a positive test result. Although 20% of plasma samples collected from 102 female kangaroos were positive for T. gondii and 18% were positive for N. caninum, we found no association between positive results and reproductive performance. Further study will be required to clarify if, and under what circumstances, T. gondii and N. caninum are pathogenic to macropod marsupials.

  3. Acute onset of encephalomyelitis with atypical lesions associated with dual infection of Sarcocystis neurona and Toxoplasma gondii in a dog.

    Science.gov (United States)

    Gerhold, Richard; Newman, Shelley J; Grunenwald, Caroline M; Crews, Amanda; Hodshon, Amy; Su, Chunlei

    2014-10-15

    A two-year-old male, neutered, basset hound-beagle mix with progressive neurological impairment was examined postmortem. Grossly, the dog had multiple raised masses on the spinal cord between nerve roots. Microscopically, the dog had protozoal myeloencephalitis. Toxoplasma gondii and Sarcocystis neurona were detected in the CNS by immunohistochemistry and polymerase chain reaction (PCR). Sarcocysts in formalin-fixed muscle were negative for Sarcocystis by PCR. Banked serum was negative for T. gondii using the modified agglutination test, suggesting an acute case of T. gondii infection or immunosuppression; however, no predisposing immunosuppressive diseases, including canine distemper, were found. To the authors' knowledge, this is the first report of dual T. gondii and S. neurona infection in a dog. Published by Elsevier B.V.

  4. Toxoplasma gondii antibodies in wild rodents and marsupials from the Atlantic Forest, state of São Paulo, Brazil

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    Solange Maria Gennari

    Full Text Available Toxoplasma gondii is a protozoan parasite that infects a large spectrum of warm-blooded animals, including humans. Small rodents and marsupials play an important role in the epidemiology of T. gondii because they are sources of infection for domestic and feral cats. Serum samples from 151 rodents and 48 marsupials, captured in the Atlantic Forest, São Paulo State, southeastern Brazil, were analyzed for the presence of T. gondii antibodies. Antibodies detected by the modified agglutination test (MAT ≥ 25 were found in 8.6% (13/151 of the rodents and 10.4% (5/48 of the marsupials, with titers ranging from 25 to 6400 and from 25 to 3200, respectively for the rodents and marsupials. Three of the eight species of rodents (Akodon spp., Oligoryzomys nigripesand Rattus norvegicus, and one from the four marsupial species (Didelphis aurita presented positive animals. T. gondii was described for the first time in the rodent Oligoryzomys nigripes.

  5. Infection of male rats with Toxoplasma gondii induces effort-aversion in a T-maze decision-making task.

    Science.gov (United States)

    Tan, Donna; Vyas, Ajai

    2016-03-01

    Rats chronically infected with protozoan Toxoplasma gondii exhibit greater delay aversion in an inter-temporal task. Moreover T. gondii infection also results in dendritic atrophy of basolateral amygdala neurons. Basolateral amygdala is reported to bias decision making towards greater effortful alternatives. In this context, we report that T. gondii increases effort aversion in infected male rats. This host-parasite association has been widely studied in the context of loss of innate fear in the infected males. It is suggested that reduced fear towards predators reflects a parasitic behavioral manipulation to enhance trophic transmission of T. gondii. Observations reported here extend this paradigm away from a monolithic change in fear and towards a multi-dimensional change in decision making. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. Metabolic Cooperation of Glucose and Glutamine Is Essential for the Lytic Cycle of Obligate Intracellular Parasite Toxoplasma gondii*

    OpenAIRE

    Nitzsche, Richard; Zagoriy, Vyacheslav; Lucius, Richard; Gupta, Nishith

    2015-01-01

    Toxoplasma gondii is a widespread protozoan parasite infecting nearly all warm-blooded organisms. Asexual reproduction of the parasite within its host cells is achieved by consecutive lytic cycles, which necessitates biogenesis of significant energy and biomass. Here we show that glucose and glutamine are the two major physiologically important nutrients used for the synthesis of macromolecules (ATP, nucleic acid, proteins, and lipids) in T. gondii, and either of them is sufficient to ensure ...

  7. Influence of Toxoplasma Gondii Infection on Symptoms and Signs of Premenstrual Syndrome: A Cross-sectional Study

    Science.gov (United States)

    Alvarado-Esquivel, Cosme; Sánchez-Anguiano, Luis Francisco; Hernández-Tinoco, Jesús; Pérez-Álamos, Alma Rosa; Rico-Almochantaf, Yazmin del Rosario; Estrada-Martínez, Sergio; Vaquera-Enriquez, Raquel; Díaz-Herrera, Arturo; Ramos-Nevarez, Agar; Sandoval-Carrillo, Ada Agustina; Salas-Pacheco, José Manuel; Cerrillo-Soto, Sandra Margarita; Antuna-Salcido, Elizabeth Irasema; Liesenfeld, Oliver; Guido-Arreola, Carlos Alberto

    2016-01-01

    Infection with Toxoplasma gondii in brain may cause some symptoms that resemble those in women with premenstrual syndrome. To determine the association of T. gondii infection with symptoms and signs of premenstrual syndrome, we examined 489 women aged 30–40 years old. Sera of participants were analyzed for the presence of anti-Toxoplasma IgG and IgM antibodies using enzyme-linked immunoassays (EIA) and T. gondii DNA by polymerase chain reaction (PCR). Anti-T. gondii IgG antibodies were found in 38 (7.8%) of the women studied. Anti-T. gondii IgM antibodies were found in 13 (34.2%) of the 38 IgG seropositive women. Logistic regression showed two variables associated with seropositivity to T. gondii: presence of diarrhea (odds ratio [OR] = 6.10; 95% confidence interval [CI]: 1.37–27.85; P = 0.01) and weight gain (OR = 2.89; 95% CI: 1.37–6.07; P = 0.005), and two variables associated with high (>150 IU/ml) levels of IgG against T. gondii: presence of diarrhea (OR = 7.40; 95% CI: 1.79–30.46; P = 0.006) and abdominal inflammation (OR = 3.38; 95% CI: 1.13–10.10; P = 0.02). Positivity to EIA IgG and PCR was positively associated with obesity and negatively associated with joint pain by bivariate analysis. Our study for the first time reveals a potential association of T. gondii infection with clinical manifestations of premenstrual syndrome. PMID:27980858

  8. Prevalence of Toxoplasma gondii antibodies in domestic donkeys (Equus asinus) in Durango, Mexico slaughtered for human consumption

    OpenAIRE

    Alvarado-Esquivel, Cosme; Alvarado-Esquivel, Domingo; Dubey, Jitender P

    2015-01-01

    Background Nothing is known about Toxoplasma gondii prevalence in donkeys in Mexico. Meat from donkey is consumed by humans in Mexico and also exported to other countries. We sought to determine the presence of antibodies against T. gondii in 239 domestic donkeys (Equus asinus) for slaughter in Durango, Mexico using the modified agglutination test (MAT). Donkeys were sampled in four premises (trade centers) where donkeys were gather for shipment to abattoirs in other Mexican states. Results A...

  9. Hsp47 and cyclophilin B traverse the endoplasmic reticulum with procollagen into pre-Golgi intermediate vesicles. A role for Hsp47 and cyclophilin B in the export of procollagen from the endoplasmic reticulum.

    Science.gov (United States)

    Smith, T; Ferreira, L R; Hebert, C; Norris, K; Sauk, J J

    1995-08-04

    Hsp47 and cyclophilin B (CyPB) are residents of the endoplasmic reticulum (ER). Both of these proteins are closely associated with polysome-associated alpha 1(I) procollagen chains. Hsp47 possesses chaperone properties early during the translation of procollagen while the cis/trans-isomerase properties of CyPB facilitate procollagen folding. In this report, we further investigate the interaction of these proteins with procollagen I during export from the ER. To inhibit vesicular budding and retain procollagen within the ER, cells were treated with the heterotrimeric G protein inhibitor mastoparan or calphostin C, a specific inhibitor of diacylglycerol/phorbol ester binding proteins. To arrest procollagen in pre-Golgi intermediate vesicles, cells were treated with guanosine 5'-3-O-(thio)triphosphate. Pulse-chase experiments of cells labeled with [35S]methionine followed by immunoprecipitation during the chase period with anti-procollagen, anti-Hsp47, and anti-CyPB antibodies were performed to reveal the relationship between Hsp47/CyPB/procollagen I. The distribution of procollagen, Hsp47, and CyPB to the ER and/or pre-Golgi vesicles was verified by immunofluorescence. Hsp47 and CyPB remained associated with procollagen retained within the ER. Hsp47 and CyPB were also associated with procollagen exported from the ER into pre-Golgi intermediate vesicles. Treatment of cells with cyclosporin A diminished the levels of CyPB bound to procollagen and diminished the rate of Hsp47 released from procollagen and the rate of procollagen secretion, suggesting that Hsp47 release from procollagen may be driven by helix formation. Also, these studies suggest that Hsp47 may resemble protein disulfide isomerase and possess both chaperone and anti-chaperone properties. During translation, high levels of Hsp47 are seen to limit protein aggregation and facilitate chain registration. Later, Hsp47 and/or CyPB and protein disulfide isomerase act as anti-chaperones and provide the basis for

  10. Diagnosis and isolation of Toxoplasma gondii in horses from Brazilian slaughterhouses Diagnóstico e isolamento de Toxoplasma gondii em equídeos de frigoríficos brasileiros

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    Fernanda Evers

    Full Text Available This study aimed to investigate anti-Toxoplasma gondii antibodies and to isolate the parasite from the brains of horses processed at slaughterhouses in Brazil. We collected brain and blood samples from 398 horses of various ages, from six Brazilian states. Serum samples were evaluated by indirect fluorescent antibody test (IFAT cut-off titre ≥ 1:64, and brains were submitted to mouse bioassay. Among the 398 horses, positivity for T. gondii was identified in 46 (11.6% by IFAT and in 14 (3.5% by mouse bioassay. In 12 of those 14 bioassays, mice were positive only by IFAT (cut-off titre ≥ 1:16, T. gondii being isolated in the remaining two. Using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP analysis of 18S rDNA to differentiate among T. gondii, Neospora caninum, and Sarcocystis neurona, we found that two of the 14 brains were positive for T. gondii only. For genotyping of the two isolates and the PCR-positive brain, we performed PCR-RFLP based on 13 markers, and SAG2 all samples were Toxoplasma gondii type I. Collectively, IFAT of horse sera and mouse bioassay identified positivity in 60 (15% of the samples. Our results show that some horses sent to slaughter in Brazil have been exposed to T. gondii.O objetivo do estudo foi investigar anticorpos anti-Toxoplasma gondii e isolar o parasita do cérebro de equídeos abatidos em matadouros-frigoríficos no Brasil. Colheram-se amostras de 398 cérebros e sangue de equídeos machos e fêmeas de idades variadas, provenientes de seis estados brasileiros. As amostras de soro foram avaliadas pelo teste de imunofluorescência indireta (IFI para T. gondii (ponto de corte ≥ 64, e os fragmentos de cérebros foram submetidos ao bioensaio em camundongos. Por meio da IFI, 46 (11,6% equídeos foram soropositivos. Pelo bioensaio em camundongos, 14 (3,5% cérebros de equídeos testados foram positivos. Em doze dos bioensaios, os camundongos foram positivos somente pela IFI (ponto

  11. Investigating seagrass in Toxoplasma gondii transmission in Florida (Trichechus manatus latirostris) and Antillean (T. m. manatus) manatees

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    Wyrosdick, Heidi M; Gerhold, Richard; Su, Chunlei; Mignucci-Giannoni, Antonio A.; Bonde, Robert K.; Chapman, Alycia; Riviera-Perez, Carla; Martinez, Jessica; Miller, Debra L.

    2017-01-01

    Toxoplasma gondii is a feline protozoan reported to cause morbidity and mortality in manatees and other marine mammals. Given the herbivorous nature of manatees, ingestion of oocysts from contaminated water or seagrass is presumed to be their primary mode of infection. The objectives of this study were to investigate oocyst contamination of seagrass beds in Puerto Rico and determine the seroprevalence of T. gondii in Antillean (Trichechus manatus manatus) and Florida (T. m. latirostris) manatees. Sera or plasma from Antillean (n = 5) and Florida (n = 351) manatees were tested for T. gondii antibodies using the modified agglutination test. No T. gondii DNA was detected via PCR in seagrass samples (n = 33) collected from Puerto Rico. Seroprevalence was 0%, suggesting a lower prevalence of T. gondii in these manatee populations than previously reported. This was the first study to investigate the potential oocyst contamination of the manatee diet, and similar studies are important for understanding the epidemiology of T. gondii in herbivorous marine mammals.

  12. Toxoplasma gondii Infection in Mice Impairs Long-Term Fear Memory Consolidation through Dysfunction of the Cortex and Amygdala.

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    Ihara, Fumiaki; Nishimura, Maki; Muroi, Yoshikage; Mahmoud, Motamed Elsayed; Yokoyama, Naoaki; Nagamune, Kisaburo; Nishikawa, Yoshifumi

    2016-10-01

    Chronic infection with Toxoplasma gondii becomes established in tissues of the central nervous system, where parasites may directly or indirectly modulate neuronal function. Epidemiological studies have revealed that chronic infection in humans is a risk factor for developing mental diseases. However, the mechanisms underlying parasite-induced neuronal dysfunction in the brain remain unclear. Here, we examined memory associated with conditioned fear in mice and found that T. gondii infection impairs consolidation of conditioned fear memory. To examine the brain pathology induced by T. gondii infection, we analyzed the parasite load and histopathological changes. T. gondii infects all brain areas, yet the cortex exhibits more severe tissue damage than other regions. We measured neurotransmitter levels in the cortex and amygdala because these regions are involved in fear memory expression. The levels of dopamine metabolites but not those of dopamine were increased in the cortex of infected mice compared with those in the cortex of uninfected mice. In contrast, serotonin levels were decreased in the amygdala and norepinephrine levels were decreased in the cortex and amygdala of infected mice. The levels of cortical dopamine metabolites were associated with the time spent freezing in the fear-conditioning test. These results suggest that T. gondii infection affects fear memory through dysfunction of the cortex and amygdala. Our findings provide insight into the mechanisms underlying the neurological changes seen during T. gondii infection. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  13. Use of filter papers to determine seroprevalence of Toxoplasma gondii among hunted ungulates in remote Peruvian Amazon.

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    Aston, Emily J; Mayor, Pedro; Bowman, Dwight D; Mohammed, Hussni O; Liotta, Janice L; Kwok, Oliver; Dubey, J P

    2014-04-01

    Toxoplasmosis is a zoonosis caused by the protozoan Toxoplasma gondii, and it is found worldwide. To determine whether ungulates are reservoirs of T. gondii in an isolated and remote region of the northeastern Peruvian Amazon, antibodies to T. gondii were determined in 5 species of ungulates by the modified agglutination test (MAT). These animals were hunted by subsistence hunters along the Yavarí-Mirín River, in the northeastern Peruvian Amazon. Blood samples were collected by hunters on filter papers. For determination of T. gondii antibodies, blood was eluted from filter papers, and a titer of 1:25 was considered indicative of exposure to T. gondii. Antibodies to T. gondii were found in 26 (31.0%) peccaries (Pecari tajacu, Tayassu pecari), six (17.1%) brocket deer (Mazama americana, Mazama gouazoubira), and four (40.0%) lowland tapir (Tapirus terrestris). We also introduced a modification to the MAT protocol that allows the extraction of fluid samples from several types of laboratory-grade filter paper, thus enabling researchers to easily adapt their approaches to the materials presented to them.

  14. Use of filter papers to determine seroprevalence of Toxoplasma gondii among hunted ungulates in remote Peruvian Amazon☆

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    Aston, Emily J.; Mayor, Pedro; Bowman, Dwight D.; Mohammed, Hussni O.; Liotta, Janice L.; Kwok, Oliver; Dubey, J.P.

    2013-01-01

    Toxoplasmosis is a zoonosis caused by the protozoan Toxoplasma gondii, and it is found worldwide. To determine whether ungulates are reservoirs of T. gondii in an isolated and remote region of the northeastern Peruvian Amazon, antibodies to T. gondii were determined in 5 species of ungulates by the modified agglutination test (MAT). These animals were hunted by subsistence hunters along the Yavarí-Mirín River, in the northeastern Peruvian Amazon. Blood samples were collected by hunters on filter papers. For determination of T. gondii antibodies, blood was eluted from filter papers, and a titer of 1:25 was considered indicative of exposure to T. gondii. Antibodies to T. gondii were found in 26 (31.0%) peccaries (Pecari tajacu, Tayassu pecari), six (17.1%) brocket deer (Mazama americana, Mazama gouazoubira), and four (40.0%) lowland tapir (Tapirus terrestris). We also introduced a modification to the MAT protocol that allows the extraction of fluid samples from several types of laboratory-grade filter paper, thus enabling researchers to easily adapt their approaches to the materials presented to them. PMID:24918073

  15. Toxoplasma gondii Infection and Mixed Anxiety and Depressive Disorder: A Case-Control Seroprevalence Study in Durango, Mexico.

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    Alvarado-Esquivel, Cosme; Sanchez-Anguiano, Luis Francisco; Hernandez-Tinoco, Jesus; Berumen-Segovia, Luis Omar; Torres-Prieto, Yazmin Elizabeth; Estrada-Martinez, Sergio; Perez-Alamos, Alma Rosa; Ortiz-Jurado, Maria Nalleli; Molotla-de-Leon, Gabriel; Beristain Garcia, Isabel; Rabago-Sanchez, Elizabeth; Liesenfeld, Oliver

    2016-07-01

    The parasite Toxoplasma gondii (T. gondii) may invade the brain and might induce behavioral changes. We sought to determine the association of T. gondii infection and mixed anxiety and depressive disorder. Through an age- and gender-matched case-control seroprevalence study, we examined 65 patients suffering from mixed anxiety and depressive disorder (WHO ICD-10 code: F41.2) attending in a public hospital of mental health and 260 control subjects without this disorder from the general population. Sera of participants were analyzed for anti-Toxoplasma IgG and IgM antibodies using enzyme-linked immunoassays. Fifteen (23.1%) of the 65 patients and 18 (6.9%) of the 260 controls had anti-T. gondii IgG antibodies (odds ratio (OR): 4.03; 95% confidence interval (CI): 1.90 - 8.53; P 150 IU/mL) anti-T. gondii IgG levels was similar in cases and controls (OR: 0.25; 95% CI: 0.05 - 1.06; P = 0.05). Seroprevalence was similar in male cases and controls (P = 1.0); however, seroprevalence was significantly higher in female cases than in female controls (OR: 7.08; 95% CI: 2.83 - 17.67; P mixed anxiety and depressive disorder. Further research to confirm this association and to determine the seroepidemiology of T. gondii infection in patients with this disorder is needed.

  16. Sequence Variation in Rhoptry Neck Protein 10 Gene among Toxoplasma gondii Isolates from Different Hosts and Geographical Locations.

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    Zhao, Yu; Zhou, Donghui; Chen, Jia; Sun, Xiaolin

    2017-01-01

    Toxoplasma gondii, as a eukaryotic parasite of the phylum Apicomplexa, can infect almost all the warm-blooded animals and humans, causing toxoplasmosis. Rhoptry neck proteins (RONs) play a key role in the invasion process of T. gondii and are potential vaccine candidate molecules against toxoplasmosis. The present study examined sequence variation in the rhoptry neck protein 10 (TgRON10) gene among 10 T. gondii isolates from different hosts and geographical locations from Lanzhou province during 2014, and compared with the corresponding sequences of strains ME49 and VEG obtained from the ToxoDB database, using polymerase chain reaction (PCR) amplification, sequence analysis, and phylogenetic reconstruction by Bayesian inference (BI) and maximum parsimony (MP). Analysis of all the 12 TgRON10 genomic and cDNA sequences revealed 7 exons and 6 introns in the TgRON10 gDNA. The complete genomic sequence of the TgRON10 gene ranged from 4759 bp to 4763 bp, and sequence variation was 0-0.6% among the 12 T. gondii isolates, indicating a low sequence variation in TgRON10 gene. Phylogenetic analysis of TgRON10 sequences showed that the cluster of the 12 T. gondii isolates was not completely consistent with their respective genotypes. TgRON10 gene is not a suitable genetic marker for the differentiation of T. gondii isolates from different hosts and geographical locations, but may represent a potential vaccine candidate against toxoplasmosis, worth further studies.

  17. Stray dogs as indicators of Toxoplasma gondii distributed in the environment: the first report across an urban-rural gradient in China

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    Yan Chao

    2012-01-01

    Full Text Available Abstract Background Toxoplasmosis is an important parasitic zoonosis caused by the protozoan Toxoplasma gondii that is distributed world-wide and infects a variety of hosts. However, the prevalence of T. gondii in the environment (such as soil, water and food is largely unknown. Due to the technical difficulty in oocyst counting directly, an alternative assay using the serologic status of T. gondii in free-living animals, such as stray or free-living dogs, as an indicator, can be used to evaluate environmental contamination indirectly, as they are exposed to the same risk of infection as humans and other animals. Results In the present study, 231 stray or free-living dogs across an urban-rural gradient were examined to assess the frequency of T. gondii in the environment. Specific antibodies to T. gondii were found in 93 dogs (40.3% by enzyme-linked immunosorbent assay (ELISA, and no statistically significant differences were observed in seroprevalences of T. gondii between urban dogs (38.7% and rural dogs (41% (p > 0.05. Conclusions A high seroprevalence of T. gondii in stray or free-living dogs in the present study indicates that there would be a wide distribution and a constant infection pressure of T. gondii across an urban-rural gradient, and the oocysts of T. gondii in the environment would be an important source of infection for humans and other animals both in urban and rural areas in China.

  18. High prevalence and genotypes of Toxoplasma gondii isolated from goats, from a retail meat store, destined for human consumption in the USA.

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    Dubey, J P; Rajendran, C; Ferreira, L R; Martins, J; Kwok, O C H; Hill, D E; Villena, I; Zhou, H; Su, C; Jones, J L

    2011-07-01

    Little information is available concerning the presence of viable Toxoplasma gondii in tissues of goats worldwide. In the present study, hearts of 234 goats obtained from a local USA grocery store were examined for T. gondii infection. Blood clot or fluid removed from each heart was tested for antibodies to T. gondii by using the modified agglutination test (MAT). Antibodies to T. gondii were found in 125 (53.4%) of 234 goats, with titers of 1:5 in 20, 1:10 in 44, 1:20 in 16, 1:40 in five, 1:160 in five, 1:320 in five, and 1:640 or higher in 30 goats. Hearts of 112 goats (46 goats goats 1:10 or higher) were used for isolation of viable T. gondii by bioassays in mice. For bioassays, 50 g of the myocardium were digested in an acid pepsin solution and the digest inoculated into mice; the recipient mice were examined for T. gondii infection. Toxoplasma gondii was isolated from 29 goats; from hearts of one of 46 with titers of goat strains. Taken together, these results indicate high parasite prevalence and moderate genetic diversity of T. gondii in goats, which have important implications in public health. We believe this is the first genetic analysis of T. gondii isolates from goats in the USA. Published by Elsevier Ltd.

  19. Decarbonylated cyclophilin A Cpr1 protein protects Saccharomyces cerevisiae KNU5377Y when exposed to stress induced by menadione.

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    Kim, Il-Sup; Jin, Ingnyol; Yoon, Ho-Sung

    2011-01-01

    Cyclophilins are conserved cis-trans peptidyl-prolyl isomerase that are implicated in protein folding and function as molecular chaperones. The accumulation of Cpr1 protein to menadione in Saccharomyces cerevisiae KNU5377Y suggests a possibility that this protein may participate in the mechanism of stress tolerance. Stress response of S. cerevisiae KNU5377Y cpr1Δ mutant strain was investigated in the presence of menadione (MD). The growth ability of the strain was confirmed in an oxidant-supplemented medium, and a relationship was established between diminishing levels of cell rescue enzymes and MD sensitivity. The results demonstrate the significant effect of CPR1 disruption in the cellular growth rate, cell viability and morphology, and redox state in the presence of MD and suggest the possible role of Cpr1p in acquiring sensitivity to MD and its physiological role in cellular stress tolerance. The in vivo importance of Cpr1p for antioxidant-mediated reactive oxygen species (ROS) neutralization and chaperone-mediated protein folding was confirmed by analyzing the expression changes of a variety of cell rescue proteins in a CPR1-disrupted strain. The cpr1Δ to the exogenous MD showed reduced expression level of antioxidant enzymes, molecular chaperones, and metabolic enzymes such as nicotinamide adenine dinucleotide phosphate (NADPH)- or adenosine triphosphate (ATP)-generating systems. More importantly, it was shown that cpr1Δ mutant caused imbalance in the cellular redox homeostasis and increased ROS levels in the cytosol as well as mitochondria and elevated iron concentrations. As a result of excess ROS production, the cpr1Δ mutant provoked an increase in oxidative damage and a reduction in antioxidant activity and free radical scavenger ability. However, there was no difference in the stress responses between the wild-type and the cpr1Δ mutant strains derived from S. cerevisiae BY4741 as a control strain under the same stress. Unlike BY4741, KNU5377Y Cpr1

  20. Octasaccharide is the minimal length unit required for efficient binding of cyclophilin B to heparin and cell surface heparan sulphate.

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    Vanpouille, Christophe; Denys, Agnès; Carpentier, Mathieu; Pakula, Rachel; Mazurier, Joël; Allain, Fabrice

    2004-09-01

    Cyclophilin B (CyPB) is a heparin-binding protein first identified as a receptor for cyclosporin A. In previous studies, we reported that CyPB triggers chemotaxis and integrin-mediated adhesion of T-lymphocytes by way of interaction with two types of binding sites. The first site corresponds to a signalling receptor; the second site has been identified as heparan sulphate (HS) and appears crucial to induce cell adhesion. Characterization of the HS-binding unit is critical to understand the requirement of HS in pro-adhesive activity of CyPB. By using a strategy based on gel mobility shift assays with fluorophore-labelled oligosaccharides, we demonstrated that the minimal heparin unit required for efficient binding of CyPB is an octasaccharide. The mutants CyPB(KKK-) [where KKK- refers to the substitutions K3A(Lys3-->Ala)/K4A/K5A] and CyPB(DeltaYFD) (where Tyr14-Phe-Asp16 has been deleted) failed to interact with octasaccharides, confirming that the Y14FD16 and K3KK5 clusters are required for CyPB binding. Molecular modelling revealed that both clusters are spatially arranged so that they may act synergistically to form a binding site for the octasaccharide. We then demonstrated that heparin-derived octasaccharides and higher degree of polymerization oligosaccharides inhibited the interaction between CyPB and fluorophore-labelled HS chains purified from T-lymphocytes, and strongly reduced the HS-dependent pro-adhesive activity of CyPB. However, oligosaccharides or heparin were unable to restore adhesion of heparinase-treated T-lymphocytes, indicating that HS has to be present on the cell membrane to support the pro-adhesive activity of CyPB. Altogether, these results demonstrate that the octasaccharide is likely to be the minimal length unit required for efficient binding of CyPB to cell surface HS and consequent HS-dependent cell responses.

  1. A comparative study of small RNAs in Toxoplasma gondii of distinct genotypes

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    Wang Jielin

    2012-09-01

    Full Text Available Abstract Background Toxoplasma gondii is an intracellular parasite with a significant impact on human health. Inside the mammalian and avian hosts, the parasite can undergo rapid development or remain inactive in the cysts. The mechanism that regulates parasite proliferation has not been fully understood. Small noncoding RNAs (sncRNA such as microRNAs (miRNAs are endogenous regulatory factors that can modulate cell differentiation and development. It is anticipated that hundreds of miRNAs regulate the expression of thousands of genes in a single organism. SncRNAs have been identified in T. gondii, however the profiles of sncRNAs expression and their potential regulatory function in parasites of distinct genotypes has largely been unknown. Methods The transcription profiles of miRNAs in the two genetically distinct strains, RH and ME49, of T. gondii were investigated and compared by a high-through-put RNA sequencing technique and systematic bioinformatics analysis. The expression of some of the miRNAs was confirmed by Northern blot analysis. Results 1,083,320 unique sequences were obtained. Of which, 17 conserved miRNAs related to 2 metazoan miRNA families and 339 novel miRNAs were identified. A total of 175 miRNAs showed strain-specific expression, of which 155 miRNAs were up-regulated in RH strain and 20 miRNAs were up-regulated in ME49 strain. Strain-specific expression of miRNAs in T. gondii could be due to activation of specific genes at different genomic loci or due to arm-switching of the same pre-miRNA duplex. Conclusions Evidence for the differential expression of miRNAs in the two genetically distinct strains of T. gondii has been identified and defined. MiRNAs of T. gondii are more species-specific as compared to other organisms, which can be developed as diagnostic biomarkers for toxoplasmosis. The data also provide a framework for future studies on RNAi-dependent regulatory mechanisms in the zoonotic parasite.

  2. Seroepidemiology of Toxoplasma gondii in zoo animals in selected zoos in the midwestern United States.

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    de Camps, Silvia; Dubey, J P; Saville, W J A

    2008-06-01

    Toxoplasma gondii infections in zoo animals are of interest because many captive animals die of clinical toxoplasmosis and because of the potential risk of exposure of children and elderly to T. gondii oocysts excreted by cats in the zoos. Seroprevalence of T. gondii antibodies in wild zoo felids, highly susceptible zoo species, and feral cats from 8 zoos of the midwestern United States was determined by using the modified agglutination test (MAT). A titer of 1:25 was considered indicative of T. gondii exposure. Among wild felids, antibodies to T. gondii were found in 6 (27.3%) of 22 cheetahs (Acynonyx jubatus jubatus), 2 of 4 African lynx (Caracal caracal), 1 of 7 clouded leopards (Neofelis nebulosa), 1 of 5 Pallas cats (Otocolobus manul), 12 (54.5%) of 22 African lions (Panthera leo), 1 of 1 jaguar (Panthera onca), 1 of 1 Amur leopard (Panthera pardus orientalis), 1 of 1 Persian leopard (Panthera pardus saxicolor), 5 (27.8%) of 18 Amur tigers (Panthera tigris altaica), 1 of 4 fishing cats (Prionailurus viverrinus), 3 of 6 pumas (Puma concolor), 2 of 2 Texas pumas (Puma concolor stanleyana), and 5 (35.7%) of 14 snow leopards (Uncia uncia). Antibodies were found in 10 of 34 feral domestic cats (Felis domesticus) trapped in 3 zoos. Toxoplasma gondii oocysts were not found in any of the 78 fecal samples from wild and domestic cats. Among the macropods, antibodies were detected in 1 of 3 Dama wallabies (Macropus eugenii), 1 of 1 western grey kangaroo (Macropus fuliginosus), 1 of 2 wallaroos (Macropus robustus), 6 of 8 Bennett's wallabies (Macropus rufogriseus), 21 (61.8%) of 34 red kangaroos (Macropus rufus), and 1 of 1 dusky pademelon (Thylogale brunii). Among prosimians, antibodies were detected in 1 of 3 blue-eyed black lemurs (Eulemur macaco flavifrons), 1 of 21 ring-tailed lemurs (Lemur catta), 2 of 9 red-ruffed lemurs (Varecia variegata rubra), and 2 of 4 black- and white-ruffed lemurs (Varecia variegata variegata). Among the avian species tested, 2 of 3 bald

  3. Toxoplasma gondii: prevalence and characterization of new genotypes in free-range chickens from south Brazil.

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    Vieira, Fernando Emmanuel Gonçalves; Sasse, João Pedro; Minutti, Ana Flávia; Miura, Ana Carolina; de Barros, Luiz Daniel; Cardim, Sergio Tosi; Martins, Thais Agostinho; de Seixas, Mércia; Yamamura, Milton Issashi; Su, Chunlei; Garcia, João Luis

    2018-03-01

    Toxoplasma gondii is an intracellular parasite that can infect all warm-blooded animals including humans. Recent studies showed that T. gondii strains from South America are genetically diverse. The present work aimed to determine T. gondii prevalence in free-ranging chicken in northwest Parana state in Brazil by two serological tests, to isolate the parasites from seropositive chickens and to genotype the isolates. Antibodies to T. gondii in 386 serum samples from 24 farms were investigated by immunofluorescence antibody assay (IFA) and modified agglutination test (MAT). Samples having titers ≥ 16 were considered positive for both tests. Among the 386 serum samples, 102 (26.4%) were positive for IFA, 64 (16.6%) were positive for MAT, 47 (12.2%) were positive in both tests, and 119 (30.8%) were positive in at least one of the two tests. Brain and pool of heart, lung, and liver from the 119 seropositive chickens were used for mouse bioassay to isolate the parasites. Thirty eight (31.9%) of these seropositive chickens were considered positives in mouse bioassay and 18 isolates were obtained. The isolates were characterized by 10 PCR-RFLP genetic markers including SAG1, SAG2 (5'-3'SAG2, alt.SAG2), SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico. Results of genotyping were compared with the genotypes in ToxoDB database. It revealed ten genotypes, including ToxoDB PCR-RFLP genotypes #6 (n = 2), #19 (n = 1), #21 (n = 2), #111 (n = 2), #152 (n = 1), and #175 (n = 1) and four new types not described before. Our results confirmed a high genetic diversity of this parasite in southern Brazil and also showed that the use of two serological tests in combination can improve the chance of T. gondii isolation. More studies should be taken to determine the zoonotic potential of chickens in the transmission of T. gondii.

  4. Seroepidemiology of Toxoplasma gondii infection in drivers involved in road traffic accidents in the metropolitan area of Guadalajara, Jalisco, Mexico.

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    Galván-Ramírez, Ma de la Luz; Sánchez-Orozco, Laura Verónica; Rodríguez, Laura Rocío; Rodríguez, Saúl; Roig-Melo, Enrique; Troyo Sanromán, Rogelio; Chiquete, Erwin; Armendáriz-Borunda, Juan

    2013-10-11

    The prevalence of toxoplasmosis in the general population of Guadalajara, Mexico, is around 32%. Toxoplasmosis can cause ocular lesions and slowing of reaction reflexes. Latent toxoplasmosis has been related with traffic accidents. We aimed to assess the prevalence of anti-Toxoplasma gondii antibodies and visual impairments related with traffic accidents in drivers from the metropolitan Guadalajara. We prospectively evaluated the prevalence of IgG and IgM anti-T. gondii antibodies in 159 individuals involved in traffic accidents, and in 164 control drivers never involved in accidents. Cases of toxoplasmosis reactivation or acute infection were detected by PCR in a subset of 71 drivers studied for the presence of T. gondii DNA in blood samples. Ophthalmologic examinations were performed in drivers with IgG anti-T. gondii antibodies in search of ocular toxoplasmosis. Fifty-four (34%) traffic accident drivers and 59 (36%) controls were positive to IgG anti-T. gondii antibodies (p = 0.70). Among the 113 seropositive participants, mean anti-T. gondii IgG antibodies titers were higher in traffic accident drivers than in controls (237.9 ± 308.5 IU/ml vs. 122.9 ± 112.7 IU/ml, respectively; p = 0.01 by Student's t test, p = 0.037 by Mann-Whitney U test). In multivariate analyses, anti-T. gondii IgG antibody titers were consistently associated with an increased risk of traffic accidents, whereas age showed an inverse association. The presence of IgM-anti-T. gondii antibodies was found in three (1.9%) subjects among traffic accident drives, and in two (1.2%) controls. Three (4.2%) samples were positive for the presence of T. gondii DNA, all among seropositive individuals. No signs of ocular toxoplasmosis were found in the entire cohort. Moreover, no other ocular conditions were found to be associated with the risk of traffic accidents in a multivariate analysis. Anti-T. gondii antibody titers are associated with the risk of traffic accidents. We could not determine any

  5. Lack of Serological and Molecular Association between Toxoplasma Gondii Exposure and Obesity: A Case-Control Study

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    Alvarado-Esquivel, Cosme; Maldonado-Soto, Edith; Sanchez-Anguiano, Luis Francisco; Hernandez-Tinoco, Jesus; Ramos-Nevarez, Agar; Cerrillo-Soto, Sandra Margarita; Sandoval-Carrilo, Ada Agustina; Salas-Pacheco, Jose Manuel; Antuna-Salcido, Elizabeth Irasema; Estrada-Martinez, Sergio; Guido-Arreola, Carlos Alberto

    2017-01-01

    The association between T. gondii infection and obesity has been scantly studied. Through an age-, and gender-matched case-control study, we determined the association of T. gondii infection and obesity using serological and molecular methods. Cases included 203 persons with obesity, and controls included 203 persons without obesity. Participants were tested for the presence of anti-Toxoplasma IgG antibodies using an enzyme-linked immunoassay (EIA). IgG seropositive individuals were further tested for the presence of anti-T. gondii IgM antibodies using an EIA, and T. gondii DNA by polymerase chain reaction (PCR). Anti-T. gondii IgG antibodies were found in 16 (7.9%) of the 203 cases and in 18 (8.9%) of the 203 controls (OR=0.87; 95% CI: 0.43-1.77; P=0.72). One (6.3%) of the 16 anti-T. gondii IgG seropositive cases and 6 (33.3%) of the 18 anti-T. gondii IgG seropositive controls were positive for IgM (P=0.09). Mean body mass index (35.5 ± 4.5) in T. gondii seropositive cases was similar (P=0.57) to that (36.1 ± 4.5) found in T. gondii seronegative cases. Stratification by obesity classes (I, II, and III) did not reveal differences (P>0.05) in seroprevalences (7.8%, 7.9%, and 8.1%, respectively) or high (>150 IU/ml) IgG antibody levels (3.3%, 3.9%, and 2.7%, respectively). PCR was positive in 5 (31.3%) of 16 cases, and in 5 (27.8%) of 18 controls examined (P=1.0). We found no serological or molecular evidence of an association between T. gondii infection and obesity in people attending a public health center in the northern Mexican city of Durango. PMID:28824343

  6. High seroprevalence of Toxoplasma gondii infection in a subset of Mexican patients with work accidents and low socioeconomic status.

    Science.gov (United States)

    Alvarado-Esquivel, Cosme; Torres-Castorena, Alejandro; Liesenfeld, Oliver; Estrada-Martínez, Sergio; Urbina-Álvarez, Jesús D

    2012-01-11

    Toxoplasma gondii has been associated with reflex impairment and traffic accidents. It is unknown whether Toxoplasma infection might be associated with work accidents. Therefore, using a case-control seroprevalence study design, 133 patients with a recent work accident and 266 control subjects of the general population from the same region were examined with enzyme-linked immunoassays for the presence and levels of anti-Toxoplasma IgG antibodies and anti-Toxoplasma IgM antibodies. Socio-demographic, work, clinical and behavioral characteristics from each worker were obtained. Eleven (8.3%) of 133 patients, and 14 (5.3%) of 266 controls had anti-T. gondii IgG antibodies. Anti-T. gondii IgG levels were higher than 150 IU/ml in 8 (6%) patients and 10 (3.8%) controls. Anti-T. gondii IgM antibodies were found in one (0.8%) of the workers, and in 6 (2.3%) of the controls. No statistically significant differences in the IgG seroprevalences, frequencies of high IgG levels, and IgM seroprevalences among patients and controls were found. In contrast, a low socio-economic level in patients with work accidents was associated with Toxoplasma seropositivity (P = 0.01). Patients with work accidents and low socioeconomic status showed a significantly (OR = 3.38; 95% CI: 0.84-16.06; P = 0.04) higher seroprevalence of T. gondii infection than controls of the same socioeconomic status (15.1% vs. 5%, respectively). Multivariate analysis showed a positive association of T. gondii infection with boar meat consumption (OR = 3.04; 95% CI: 1.03-8.94; P = 0.04). In contrast, a negative association between T. gondii infection and national trips (OR = 0.40; 95% CI: 0.17-0.96; P = 0.04), sausage consumption (OR = 0.20; 95% CI: 0.05-0.68; P = 0.01), and ham consumption (OR = 0.16; 95% CI: 0.05-0.51; P = 0.002) was found. In the study described here seropositivity to T. gondii was associated to work accidents in a subset of patients with low socioeconomic status. This is the first report of an

  7. A meta-analysis of the prevalence of toxoplasma gondii igg antibodies in patients with mental disorders

    Directory of Open Access Journals (Sweden)

    Angela Dragomir1,

    2016-12-01

    Full Text Available OBJECTIVES AND BACKGROUND Toxoplasma gondii infection has been recently associated with schizophrenia and other psychiatric disorders. The aim of the present study was to evaluate the prevalence of T. gondii infection in psychiatric patients by using meta-analytical methods. MATERIALS AND METHODS By systematic research of PUBMED Database, we identified several articles on this issue. We included casecontrol studies focused on the seroprevalence of T. gondii (IgG antibodies in patients with psychiatric disorders and healthy controls, published over the past 10 years R3.2.2. free software for statistical computing and graphics was used to perform the meta-analysis. Data were pooled using a random effects model and Mantel-Haenszel method. RESULTS The PUBMED Database showed references to only seven scientific papers that investigated the prevalence of T. gondii IgG antibodies in psychiatric patients. Six hundred seventy three patients and seven hundred seventy four controls coped with the inclusion criteria and were used in our analysis. We found a significant increase of T. gondii IgG antibodies in patients with schizophrenia and other psychiatric disorders compared with controls (41.6% vs 24.54%, OR = 2.16, 95% CI = [1.45-3.21], P = .001. CONCLUSIONS An increased seroprevalence of T. gondii IgG antibodies has been reported in psychiatric patients. Our study suggests that T. gondii infection may be relevant in order to determine and understand the complex etiology of schizophrenia and other psychiatric disorders. Graphical abstract: Meta-analysis of the prevalence of T. gondii IgG antibodies in patients with mental disorders REFERENCES 1. Hamidinejat H, Ghorbanpoor M, Hosseini H, Alavi SM, Nabavi L, Jalali MH, Borojeni MP, Jafari H, Mohammadaligol S. Toxoplasma gondii infection in first-episode and inpatient individuals with schizophrenia. Int J Infect Dis. 2010;14:e978-81. doi: 10.1016/j.ijid.2010.05.018. 2. Alvarado-Esquivel C, Carrillo-Oropeza D

  8. Lack of Association Between Toxoplasma gondii Infection and Diabetes Mellitus: A Matched Case-Control Study in a Mexican Population.

    Science.gov (United States)

    Alvarado-Esquivel, Cosme; Loera-Moncivais, Nayely; Hernandez-Tinoco, Jesus; Sanchez-Anguiano, Luis Francisco; Hernandez-Madrid, Guillermina; Rabago-Sanchez, Elizabeth; Centeno-Tinoco, Maria Magdalena; Sandoval-Carrillo, Ada A; Salas-Pacheco, Jose M; Campos-Moreno, Oscar Vladimir; Antuna-Salcido, Elizabeth Irasema

    2017-06-01

    Very little is known about the association between infection with Toxoplasma gondii ( T. gondii ) and diabetes mellitus. We perform an age- and gender-matched case-control study to determine the association of T. gondii infection and diabetes mellitus. Cases included 156 patients with diabetes mellitus and 156 controls without diabetes mellitus who attended in two public clinics in Durango City, Mexico. Sera of cases and controls were tested for the presence of anti- Toxoplasma IgG and IgM antibodies using commercially available enzyme-linked fluorescence assays (ELFA). Anti- T. gondii IgG antibodies were found in 10 (6.4%) of the 156 cases and in five (3.2%) of the 156 controls (odds ratio (OR): 2.06; 95% confidence interval (CI): 0.69 - 6.19; P = 0.18). The frequency of high (> 150 IU/mL) anti- T. gondii IgG levels in seropositive cases (1/10: 10.0%) was comparable to the one (1/5: 20%) in seropositive controls (OR: 0.44; 95% CI: 0.02 - 9.03; P = 1.00). None of the 10 cases and five controls with seropositivity to anti- T. gondii IgG antibodies were positive for anti- T. gondii IgM antibodies. Stratification by gender showed similar frequencies of T. gondii infection in female cases (7/107: 6.5%) and female controls (4/107: 3.7%) (OR: 1.80; 95% CI: 0.51 - 6.34; P = 0.53), and in male cases (3/49: 6.1%) and male controls (1/49: 2.0%) (OR: 3.13; 95% CI: 0.31 - 31.19; P = 0.61). We conclude that there is not serological evidence of an association between T. gondii infection and diabetes mellitus in the studied subjects in Durango City, Mexico. Further studies to elucidate the role of T. gondii in diabetes should be conducted.

  9. Toxoplasma gondii sequesters lysosomes from mammalian hosts in the vacuolar space.

    Science.gov (United States)

    Coppens, Isabelle; Dunn, Joe Dan; Romano, Julia D; Pypaert, Marc; Zhang, Hui; Boothroyd, John C; Joiner, Keith A

    2006-04-21

    The intracellular compartment harboring Toxoplasma gondii satisfies the parasite's nutritional needs for rapid growth in mammalian cells. We demonstrate that the parasitophorous vacuole (PV) of T. gondii accumulates material coming from the host mammalian cell via the exploitation of the host endo-lysosomal system. The parasite actively recruits host microtubules, resulting in selective attraction of endo-lysosomes to the PV. Microtubule-based invaginations of the PV membrane serve as conduits for the delivery of host endo-lysosomes within the PV. These tubular conduits are decorated by a parasite coat, including the tubulogenic protein GRA7, which acts like a garrote that sequesters host endocytic organelles in the vacuolar space. These data define an unanticipated process allowing the parasite intimate and concentrated access to a diverse range of low molecular weight components produced by the endo-lysosomal system. More generally, they identify a unique mechanism for unidirectional transport and sequestration of host organelles.

  10. Clinical Toxoplasma gondii, Hammondia heydorni, and Sarcocystis spp. infections in dogs.

    Science.gov (United States)

    Dubey, J P; Ross, A D; Fritz, D

    2003-12-01

    Concurrent infections with coccidians Toxoplasma gondii, Sarcocystis spp., and a Hammondia heydorni-like parasite were identified in tissues of three littermate pups on a Kelpie dog breeding farm in Australia. In total, 20 pups in four litters had died following vaccination with an attenuated distemper virus vaccine. Toxoplasma gondii tachyzoites were identified immunohistochemically in tissues of two dogs. Sarcocystis sp. sporocysts were seen in the intestinal lamina propria of two dogs. Asexual and sexual stages of H. heydorni-like parasite were found in enterocytes of the small intestine of two dogs. Ultrastructural development of schizonts and gamonts of this parasite is described. None of the protozoa in these dogs reacted with antibodies to Neospora caninum. Feeding of uncooked tissue of sheep was considered to be the likely source of infection for these coccidians in dogs.

  11. Prevalence of agglutinating antibodies to Toxoplasma gondii and Sarcocystis neurona in beavers (Castor canadensis) from Massachusetts

    Science.gov (United States)

    Jordan, C.N.; Kaur, T.; Koenen, K.; DeStefano, S.; Zajac, A.M.; Lindsay, D.S.

    2005-01-01

    The present study examined the seroprevalence of Toxoplasma gondii and Sarcocystls neurona in a population of beavers (Castor canadensis) from Massachusetts. Sixty-two blood samples were collected during the field seasons over 3 consecutive years from different animals. Blood was collected onto filter paper and shipped to the Department of Biomedical Sciences, Virginia Tech, Blacksburg, Virginia, for parasite testing. The samples were tested at dilutions of 1:25, 1:50, and 1:100 against each parasite antigen by modified agglutination tests to determine whether antibodies to either parasite were present in the blood. Six of 62 samples (10%) were positive for T. gondii, with 2 samples having titers of 1:25 and 4 having titers of 1:50. Four of 62 samples (6%) were positive for S. neurona, with 2 samples having titers of 1:25 and 2 having titers of 1:50. ?? American Society of Pathologists 2005.

  12. Seroprevalence of Toxoplasma gondii in domestic pets from metropolitan regions of Panama

    Directory of Open Access Journals (Sweden)

    Rengifo-Herrera Claudia

    2017-01-01

    Full Text Available Toxoplasmosis is a worldwide zoonotic disease but information regarding domestic animals in Central America is scarce and fragmented. The aim of this study was to determine the seroprevalence of Toxoplasma gondii in domestic cats and dogs in different metropolitan regions of Panama. A total of 576 samples were collected; sera from 120 cats and 456 dogs were tested using a commercial indirect enzyme-linked immunosorbent assay (ELISA. The overall seroprevalence of IgG antibodies was 30.73%. There is high seroprevalence of T. gondii in cats and dogs in the metropolitan regions around the Panama Canal; however, differences between these species were not significant. Statistical analysis indicated that there are relevant variables, such as the age of animals, with a direct positive relationship with seroprevalence. None of the variables related to animal welfare (veterinary attention provided, type of dwelling, and access to green areas and drinking water were associated with seropositivity.

  13. Serologic evidence of Toxoplasma gondii infection in wild birds and mammals from southeast Brazil.

    Science.gov (United States)

    Vitaliano, Sérgio Netto; Soares, Herbert Sousa; Pena, Hilda Fáitima de Jesus; Dubey, Jitender Prakash; Gennari, Solange Maria

    2014-03-01

    In this study, serum samples of 53 wild animals from two different states from the southeast region of Brazil were analyzed for the presence of anti-Toxoplasma gondii antibodies by the modified agglutination test (MAT), with a cut-off of 1: 5 for birds and of 1: 25 for mammals. Out of the sampled animals, 27 were birds and 26 were mammals, and from this total, 83% (n = 44) were free-living animals. Antibodies were found in 13 mammals, from which 11 were free-living animals, and in five birds, all of which were free-living. In this study, T. gondii antibodies were detected in four bird species (crested seriema, Cariama cristata; buff-necked ibis, Theristicus caudatus; picazuro pigeon, Patagioenas picazuro; and burrowing owl, Athene cunicularia) and in a giant anteater (Myrmecophaga tridactyla) for the first time.

  14. Seroprevalence of Toxoplasma gondii infection in goats and sheep in Zimbabwe

    DEFF Research Database (Denmark)

    Hove, T.; Lind, Peter; Mukaratirwa, S.

    2005-01-01

    Seroprevalence rates of Toxoplasma gondii anti-antibodies in adult goats and sheep from different parts of Zimbabwe were determined. A total of 225 (67.9 %) of the 335 serum samples tested were positive for anti-T. gondii IgG antibodies with the indirect fluorescent antibody test. There were...... differences in antibody seroprevalences among communal land goats from the different agro-ecological zones (Natural regions IIb and III: 80 and 96.7%, respectively; Natural region IV: 65.9%; Natural region V: 45%; and Natural region III had a significantly higher seroprevalence than IV and V. The highest...... in sheep from a large commercial farm (10%) was significantly lower than that of sheep reared under the communal grazing system (80%). Overall, significantly higher proportions of seropositive animals had antibody titres of 1:50 (34.2% of 225) and 1:100 (44% of 225) as compared to the 9.8% and 12...

  15. Detection of Toxoplasma gondii in a free-ranging giant anteater

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    Thais Oliveira Morgado

    Full Text Available ABSTRACT: Toxoplasmosis is caused by Toxoplasma gondii, an obligatory intracellular protozoan, which establishes acute and chronic infections in birds and mammals, including humans. This note reports, for the first time, the detection and sequencing of DNA from T. gondii in the peripheral blood of a young free range giant anteater (Myrmecophaga tridactyla. For the diagnosis, the following methods were used: polymerase chain reaction (PCR and positive serology (1:800 by means of the modified agglutination test (MAT. Since this species may be consumed by humans and predated by wild felids, its importance is emphasized as a probable source of zoonotic infection, in addition to its possible participation in the infection enzootic cycle. Although, parasitemia has been confirmed in this specimen, it presented no clinical sign of infection.

  16. Frequency of Toxoplasma gondii antibodies in bovines in the state of Pernambuco, Brazil

    Directory of Open Access Journals (Sweden)

    Neurisvan Ramos Guerra

    Full Text Available Toxoplasmosis is a parasitic disease caused by Toxoplasma gondii that affects homeothermic animals, including humans. Felines are considered the definitive host of this parasite, while other animals act as intermediate hosts. The purpose of this study was to assess the frequency of anti-T. gondii IgG antibodies in bovines in the state of Pernambuco, Brazil. Serum samples (n = 427 from animals in 13 municipalities of the coastal forest/plantation region of the state were analyzed using the immunofluorescent antibody test (IFAT. The overall results revealed a prevalence rate of 16.63% (27/427. High percentages of positivity were found among animals aged 25 to 36 months (28.57%; 30/42 and in males (22.22%; 2/9. The present findings suggest that bovine toxoplasmosis is endemic in the area under study.

  17. The cyclophilin inhibitor Debio-025 shows potent anti-hepatitis C effect in patients coinfected with hepatitis C and human immunodeficiency virus.

    Science.gov (United States)

    Flisiak, Robert; Horban, Andrzej; Gallay, Philippe; Bobardt, Michael; Selvarajah, Suganya; Wiercinska-Drapalo, Alicja; Siwak, Ewa; Cielniak, Iwona; Higersberger, Jozef; Kierkus, Jarek; Aeschlimann, Christian; Grosgurin, Pierre; Nicolas-Métral, Valérie; Dumont, Jean-Maurice; Porchet, Hervé; Crabbé, Raf; Scalfaro, Pietro

    2008-03-01

    Debio-025 is an oral cyclophilin (Cyp) inhibitor with potent anti-hepatitis C virus activity in vitro. Its effect on viral load as well as its influence on intracellular Cyp levels was investigated in a randomized, double-blind, placebo-controlled study. Mean hepatitis C viral load decreased significantly by 3.6 log(10) after a 14-day oral treatment with 1200 mg twice daily (P CypB) levels in peripheral blood mononuclear cells decreased from 67 +/- 6 (standard error) ng/mg protein (baseline) to 5 +/- 1 ng/mg protein at day 15 (P CypB levels, coinciding with the decrease in hepatitis C viral load. These are the first preliminary human data supporting the hypothesis that CypB may play an important role in hepatitis C virus replication and that Cyp inhibition is a valid target for the development of anti-hepatitis C drugs.

  18. Cyclophilin A Levels Dictate Infection Efficiency of Human Immunodeficiency Virus Type 1 Capsid Escape Mutants A92E and G94D ▿

    Science.gov (United States)

    Ylinen, Laura M. J.; Schaller, Torsten; Price, Amanda; Fletcher, Adam J.; Noursadeghi, Mahdad; James, Leo C.; Towers, Greg J.

    2009-01-01

    Cyclophilin A (CypA) is an important human immunodeficiency virus type 1 (HIV-1) cofactor in human cells. HIV-1 A92E and G94D capsid escape mutants arise during CypA inhibition and in certain cell lines are dependent on CypA inhibition. Here we show that dependence on CypA inhibition is due to high CypA levels. Restricted HIV-1 is stable, and remarkably, restriction is augmented by arresting cell division. Nuclear entry is not inhibited. We propose that high CypA levels and capsid mutations combine to disturb uncoating, leading to poor infectivity, particularly in arrested cells. Our data suggest a role for CypA in uncoating the core of HIV-1 to facilitate integration. PMID:19073742

  19. Development of novel strategy for the synthesis of organometallic compounds usable as protein ligands: application to the human cyclophilin hCyp-18

    International Nuclear Information System (INIS)

    Clavaud, C.

    2006-02-01

    This thesis describes a new strategy for the development of bioactive organometallic compounds, basing on the combinatorial assembly of sub-chemical libraries (A and B) independent but complementary and able to coordinate a metallic heart M to form A-M-B complex potential ligands of biomolecules. The coordination of metals, well adapted to the production of molecular variety is usually used in medicinal chemistry, in diagnostic and therapeutic nuclear medicine. Among the useful elements, the rhenium and the technetium are metals of choice for the development of the assembly strategy because of their chemical and radiochemical properties and of the structure analogy of their complexes. This strategy was validated in vitro. The protein chosen for this purpose was the cyclophilin hCyp-18. (N.C.)

  20. Recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip for detection of Toxoplasma gondii in the environment.

    Science.gov (United States)

    Wu, Y D; Xu, M J; Wang, Q Q; Zhou, C X; Wang, M; Zhu, X Q; Zhou, D H

    2017-08-30

    Toxoplasma gondii infects all warm-blooded vertebrates, resulting in a great threat to human health and significant economic loss to the livestock industry. Ingestion of infectious oocysts of T. gondii from the environment is the major source of transmission. Detection of T. gondii oocysts by existing methods is laborious, time-consuming and expensive. The objective of the present study was to develop a recombinase polymerase amplification (RPA) method combined with a lateral flow (LF) strip for detection of T. gondii oocysts in the soil and water. The DNA of T. gondii oocysts was amplified by a pair of specific primers based on the T. gondii B1 gene over 15min at a constant temperature ranging from 30°C to 45°C using RPA. The amplification product was visualized by the lateral flow (LF) strip within 5min using the specific probe added to the RPA reaction system. The sensitivity of the established assay was 10 times higher than that of nested PCR with a lower detection limit of 0.1 oocyst per reaction, and there was no cross-reactivity with other closely related protozoan species. Fifty environmental samples were further assessed for the detection validity of the LF-RPA assay (B1-LF-RPA) and compared with nested PCR based on the B1 gene sequence. The B1-LF-RPA and nested PCR both showed that 5 out of the 50 environmental samples were positive. The B1-LF-RPA method was also proven to be sufficiently tolerant of existing inhibitors in the environment. In addition, the advantages of simple operation, speediness and cost-effectiveness make B1-LF-RPA a promising molecular detection tool for T. gondii. Copyright © 2017 Elsevier B.V. All rights reserved.