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Sample records for heat labile toxin

  1. Cistrons encoding Escherichia coli heat-labile toxin.

    PubMed Central

    Dallas, W S; Gill, D M; Falkow, S

    1979-01-01

    The structure and products of the two cistrons encoding the Escherichia coli heat-labile toxin (LT) were studied. The LT deoxyribonucleic acid (DNA) region had been isolated as part of a DNA fragment from the plasmid P307, and this fragment was joined to the cloning vector pBR313. Deletion mutations of various lengths were introduced into the LT DNA region and into the adjacent DNA sequences. Analysis of the deletions indicated that the maximum size of the LT DNA region was 1.2 x 10(6) daltons. Two proteins of 11,500 daltons and 25,500 daltons had been shown to be encoded by the LT DNA region. The functions of these LT gene products were investigated. The 11,500-dalton protein had an adsorption activity for Y-1 adrenal cells, and this protein was shown to form aggregates of four or five monomers. The 25,500-dalton protein was shown to have an adenylate cyclase-activating activity. The two cistrons encoding for each of the LT proteins have been located on a genetic map of the LT DNA region. Both cistrons are probably transcribed from the same promoter. Images PMID:383697

  2. Structure and function of cholera toxin and the related Escherichia coli heat-labile enterotoxin.

    PubMed Central

    Spangler, B D

    1992-01-01

    Cholera and the related Escherichia coli-associated diarrheal disease are important problems confronting Third World nations and any area where water supplies can become contaminated. The disease is extremely debilitating and may be fatal in the absence of treatment. Symptoms are caused by the action of cholera toxin, secreted by the bacterium Vibrio cholerae, or by a closely related heat-labile enterotoxin, produced by Escherichia coli, that causes a milder, more common traveler's diarrhea. Both toxins bind receptors in intestinal epithelial cells and insert an enzymatic subunit that modifies a G protein associated with the adenylate cyclase complex. The consequent stimulated production of cyclic AMP, or other factors such as increased synthesis of prostaglandins by intoxicated cells, initiates a metabolic cascade that results in the excessive secretion of fluid and electrolytes characteristic of the disease. The toxins have a very high degree of structural and functional homology and may be evolutionarily related. Several effective new vaccine formulations have been developed and tested, and a growing family of endogenous cofactors is being discovered in eukaryotic cells. The recent elucidation of the three-dimensional structure of the heat-labile enterotoxin has provided an opportunity to examine and compare the correlations between structure and function of the two toxins. This information may improve our understanding of the disease process itself, as well as illuminate the role of the toxin in studies of signal transduction and G-protein function. Images PMID:1480112

  3. Adhesin degradation accelerates delivery of heat-labile toxin by enterotoxigenic Escherichia coli.

    PubMed

    Roy, Koushik; Kansal, Rita; Bartels, Scott R; Hamilton, David J; Shaaban, Salwa; Fleckenstein, James M

    2011-08-26

    Many enteric pathogens, including enterotoxigenic Escherichia coli (ETEC), produce one or more serine proteases that are secreted via the autotransporter (or type V) bacterial secretion pathway. These molecules have collectively been referred to as SPATE proteins (serine protease autotransporter of the Enterobacteriaceae). EatA, an autotransporter previously identified in ETEC, possesses a functional serine protease motif within its secreted amino-terminal passenger domain. Although this protein is expressed by many ETEC strains and is highly immunogenic, its precise function is unknown. Here, we demonstrate that EatA degrades a recently characterized adhesin, EtpA, resulting in modulation of bacterial adhesion and accelerated delivery of the heat-labile toxin, a principal ETEC virulence determinant. Antibodies raised against the passenger domain of EatA impair ETEC delivery of labile toxin to epithelial cells suggesting that EatA may be an effective target for vaccine development. PMID:21757737

  4. Production of Escherichia coli heat labile toxin (LT) B subunit in soybean seed and analysis of its immunogenicity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Expression of the heat-labile toxin B subunit of enterotoxigenic Escherichia coli (LT) B was directed to the endoplasmic reticulum (ER) of soybean seed storage parenchyma cells for immunogen sequestration in de novo synthesized, ER-derived protein accretions in transgenic seed. Pentameric LTB accumu...

  5. Characterization of uterine growth response to cholera toxin in hamsters and test of heat-labile enterotoxin from Escherichia coli.

    PubMed Central

    Alleva, J J; Lamanna, C

    1984-01-01

    Cholera toxin (CT) and the heat-labile enterotoxin from Escherichia coli, when injected intraperitoneally into cycling hamsters but not rats or mice, induced a massive uterine growth similar to that normally induced by the implanting blastocyst during pregnancy. CT and heat-labile enterotoxin are the only known agents that have this action in any species. Uterine weight reached a maximal sixfold increase 48 h after injection of CT. Concurrent injection of estrogen, progesterone, and CT increased the maximal response to eightfold and eliminated differences in the response to CT injected on different days of the 4-day hamster estrous cycle. The dose response for CT, heat-labile enterotoxin, and CT plus estrogen plus progesterone was most linear (r greater than 0.93) when the logarithm of uterine weight was plotted against the dose of toxin. The hamster uterine weight response can serve as a simple, highly precise, and highly specific bioassay for CT and heat-labile enterotoxin. PMID:6386865

  6. Parenteral Adjuvant Effects of an Enterotoxigenic Escherichia coli Natural Heat-Labile Toxin Variant.

    PubMed

    Braga, Catarina J M; Rodrigues, Juliana F; Medina-Armenteros, Yordanka; Farinha-Arcieri, Luís E; Ventura, Armando M; Boscardin, Silvia B; Sbrogio-Almeida, Maria E; Ferreira, Luís C S

    2014-01-01

    Native type I heat-labile toxins (LTs) produced by enterotoxigenic Escherichia coli (ETEC) strains exert strong adjuvant effects on both antibody and T cell responses to soluble and particulate antigens following co-administration via mucosal routes. However, inherent enterotoxicity and neurotoxicity (following intra-nasal delivery) had reduced the interest in the use of these toxins as mucosal adjuvants. LTs can also behave as powerful and safe adjuvants following delivery via parenteral routes, particularly for activation of cytotoxic lymphocytes. In the present study, we evaluated the adjuvant effects of a new natural LT polymorphic form (LT2), after delivery via intradermal (i.d.) and subcutaneous (s.c.) routes, with regard to both antibody and T cell responses. A recombinant HIV-1 p24 protein was employed as a model antigen for determination of antigen-specific immune responses while the reference LT (LT1), produced by the ETEC H10407 strain, and a non-toxigenic LT form (LTK63) were employed as previously characterized LT types. LT-treated mice submitted to a four dose-base immunization regimen elicited similar p24-specific serum IgG responses and CD4(+) T cell activation. Nonetheless, mice immunized with LT1 or LT2 induced higher numbers of antigen-specific CD8(+) T cells and in vivo cytotoxic responses compared to mice immunized with the non-toxic LT derivative. These effects were correlated with stronger activation of local dendritic cell populations. In addition, mice immunized with LT1 and LT2, but not with LTK63, via s.c. or i.d. routes developed local inflammatory reactions. Altogether, the present results confirmed that the two most prevalent natural polymorphic LT variants (LT1 or LT2) display similar and strong adjuvant effects for subunit vaccines administered via i.d. or s.c. routes. PMID:24432018

  7. Monoclonal antibodies to cholera toxin with special reference to cross-reactions with Escherichia coli heat-labile enterotoxin.

    PubMed Central

    Lindholm, L; Holmgren, J; Wikstrm, M; Karlsson, U; Andersson, K; Lycke, N

    1983-01-01

    Seventy monoclonal antibodies to cholera toxin were prepared and characterized. All were of immunoglobulin G (IgG) isotypes (39 IgG1, 29 IgG2, and 2 IgG3). A total of 61 clones produced antibody directed against the B subunit, and 9 clones produced antibodies with specificity for the cholera toxin A subunit. Among both the anti-B and anti-A antibodies, there were representatives which showed full cross-reactivity with the heat-labile enterotoxin of Escherichia coli (14 clones), others which gave partial cross-reactions (12), and still others (44) which did not cross-react. Although 24 of 25 tested anti-B monoclonal antibodies could neutralize cholera toxin, none of the 9 anti-A clones had any detectable neutralizing ability. Among the anti-B antibodies, those which cross-reacted completely with E. coli heat-labile enterotoxin all had strong cholera toxin-neutralizing capacity, whereas those with lesser or no degree of cross-reactivity varied more in their neutralizing potency. The isolation of monoclonal antibodies that distinguish between enterotoxins of different bacterial origin suggests the possibility of developing immunodiagnostic methods allowing species-specific enterotoxin detection in stools of patients with diarrheal disease. PMID:6840852

  8. Differential interaction of Escherichia coli heat-labile toxin and cholera toxin with pig intestinal brush border glycoproteins depending on their ABH and related blood group antigenic determinants.

    PubMed Central

    Balanzino, L E; Barra, J L; Monferran, C G; Cumar, F A

    1994-01-01

    The ability of glycoproteins from pig intestinal brush border membranes (BBM) to bind cholera toxin (CT) or heat-labile toxins from strains of Escherichia coli isolated from human (LTh) or pig (LTp) intestines was studied. Glycoproteins capable of binding the toxins are also recognized by antibodies or lectins specific for ABO(H) blood group and related antigens. Pigs expressing A, H, or I antigenic determinants were used for comparison. The toxin-binding capacity of a glycoprotein depends on the toxin type and the blood group epitope borne by the glycoprotein. LTh and LTp preferably bound to several blood group A-active glycoproteins rather than H-active glycoproteins. By contrast, CT practically did not recognize either blood group A- or blood group H-active glycoproteins, while glycoproteins from pigs expressing I antigenic determinants were able to interact with LTh, LTp, and CT. LTh, LTp, or CT glycoprotein binding was selectively inhibited by specific lectins or monosaccharides. Affinity purification of the toxin binding brush border glycoproteins on the basis of their blood group reactivity suggests that such glycoproteins are hydrolytic enzymes. BBM from A+ pigs contain about 27 times more LTh binding sites, in addition to those recognized by CT, than an equivalent membrane preparation from H+ pigs. The present findings may help clarify some previous unclear results on LTh binding to intestinal BBM glycoproteins obtained by use of animals not typed by their ABO(H) blood group phenotype. Images PMID:7510669

  9. Allele Variants of Enterotoxigenic Escherichia coli Heat-Labile Toxin Are Globally Transmitted and Associated with Colonization Factors

    PubMed Central

    Joffr, Enrique; von Mentzer, Astrid; Abd El Ghany, Moataz; Oezguen, Numan; Savidge, Tor; Dougan, Gordon; Svennerholm, Ann-Mari

    2014-01-01

    Enterotoxigenic Escherichia coli (ETEC) is a significant cause of morbidity and mortality in the developing world. ETEC-mediated diarrhea is orchestrated by heat-labile toxin (LT) and heat-stable toxins (STp and STh), acting in concert with a repertoire of more than 25 colonization factors (CFs). LT, the major virulence factor, induces fluid secretion after delivery of a monomeric ADP-ribosylase (LTA) and its pentameric carrier B subunit (LTB). A study of ETEC isolates from humans in Brazil reported the existence of natural LT variants. In the present study, analysis of predicted amino acid sequences showed that the LT amino acid polymorphisms are associated with a geographically and temporally diverse set of 192 clinical ETEC strains and identified 12 novel LT variants. Twenty distinct LT amino acid variants were observed in the globally distributed strains, and phylogenetic analysis showed these to be associated with different CF profiles. Notably, the most prevalent LT1 allele variants were correlated with major ETEC lineages expressing CS1 + CS3 or CS2 + CS3, and the most prevalent LT2 allele variants were correlated with major ETEC lineages expressing CS5 + CS6 or CFA/I. LTB allele variants generally exhibited more-stringent amino acid sequence conservation (2 substitutions identified) than LTA allele variants (22 substitutions identified). The functional impact of LT1 and LT2 polymorphisms on virulence was investigated by measuring total-toxin production, secretion, and stability using GM1enzyme-linked immunosorbent assays (GM1-ELISA) and in silico protein modeling. Our data show that LT2 strains produce 5-fold more toxin than LT1 strains (P < 0.001), which may suggest greater virulence potential for this genetic variant. Our data suggest that functionally distinct LT-CF variants with increased fitness have persisted during the evolution of ETEC and have spread globally. PMID:25404692

  10. Expression and immunogenicity of enterotoxigenic Escherichia coli heat-labile toxin B subunit in transgenic rice callus.

    PubMed

    Kim, Tae-Geum; Kim, Bang-Geul; Kim, Mi-Young; Choi, Jae-Kwon; Jung, Eun-Sun; Yang, Moon-Sik

    2010-01-01

    Enterotoxigenic Escherichia coli is one of the leading causes of diarrhea in developing countries, and the disease may be fatal in the absence of treatment. Enterotoxigenic E. coli heat-labile toxin B subunit (LTB) can be used as an adjuvant, as a carrier of fused antigens, or as an antigen itself. The synthetic LTB (sLTB) gene, optimized for plant codon usage, has been introduced into rice cells by particle bombardment-mediated transformation. The integration and expression of the sLTB gene were observed via genomic DNA PCR and western blot analysis, respectively. The binding activity of LTB protein expressed in transgenic rice callus to G(M1)-ganglioside, a receptor for biologically active LTB, was confirmed by G(M1)-ELISA. Oral inoculation of mice with lyophilized transgenic rice calli containing LTB generated significant IgG antibody titers against bacterial LTB, and the sera of immunized mice inhibited the binding of bacterial LTB to G(M1)-ganglioside. Mice orally immunized with non-transgenic rice calli failed to generate detectable anti-LTB IgG antibody titers. Mice immunized with plant-produced LTB generated higher IgG1 antibody titers than IgG2a, indicating a Th2-type immune response. Mice orally immunized with lyophilized transgenic rice calli containing LTB elicited higher fecal IgA antibody titers than mice immunized with non-transgenic rice calli. These experimental results demonstrate that LTB proteins produced in transgenic rice callus and given to mice by oral administration induce humoral and secreted antibody immune responses. We suggest that transgenic rice callus may be suitable as a plant-based edible vaccine to provide effective protection against enterotoxigenic E. coli heat-labile toxin. PMID:19657748

  11. Comparative Adjuvant Effects of Type II Heat-Labile Enterotoxins in Combination with Two Different Candidate Ricin Toxin Vaccine Antigens.

    PubMed

    Vance, David J; Greene, Christopher J; Rong, Yinghui; Mandell, Lorrie M; Connell, Terry D; Mantis, Nicholas J

    2015-12-01

    Type II heat-labile enterotoxins (HLTs) constitute a promising set of adjuvants that have been shown to enhance humoral and cellular immune responses when coadministered with an array of different proteins, including several pathogen-associated antigens. However, the adjuvant activities of the four best-studied HLTs, LT-IIa, LT-IIb, LT-IIb(T13I), and LT-IIc, have never been compared side by side. We therefore conducted immunization studies in which LT-IIa, LT-IIb, LT-IIb(T13I), and LT-IIc were coadministered by the intradermal route to mice with two clinically relevant protein subunit vaccine antigens derived from the enzymatic A subunit (RTA) of ricin toxin, RiVax and RVEc. The HLTs were tested with low and high doses of antigen and were assessed for their abilities to stimulate antigen-specific serum IgG titers, ricin toxin-neutralizing activity (TNA), and protective immunity. We found that all four HLTs tested were effective adjuvants when coadministered with RiVax or RVEc. LT-IIa was of particular interest because as little as 0.03 μg when coadministered with RiVax or RVEc proved effective at augmenting ricin toxin-specific serum antibody titers with nominal evidence of local inflammation. Collectively, these results justify the need for further studies into the mechanism(s) underlying LT-IIa adjuvant activity, with the long-term goal of evaluating LT-IIa's activity in humans. PMID:26491037

  12. Single Chain Variable Fragments Produced in Escherichia coli against Heat-Labile and Heat-Stable Toxins from Enterotoxigenic E. coli

    PubMed Central

    Andrade, Fernanda B.; Nepomuceno, Roberto; Silva, Anderson; Munhoz, Danielle D.; Yamamoto, Bruno B.; Luz, Daniela; Abreu, Patrícia A. E.; Horton, Denise S. P. Q.; Elias, Waldir P.; Ramos, Oscar H. P.; Piazza, Roxane M. F.

    2015-01-01

    Background Diarrhea is a prevalent pathological condition frequently associated to the colonization of the small intestine by enterotoxigenic Escherichia coli (ETEC) strains, known to be endemic in developing countries. These strains can produce two enterotoxins associated with the manifestation of clinical symptoms that can be used to detect these pathogens. Although several detection tests have been developed, minimally equipped laboratories are still in need of simple and cost-effective methods. With the aim to contribute to the development of such diagnostic approaches, we describe here two mouse hybridoma-derived single chain fragment variable (scFv) that were produced in E. coli against enterotoxins of ETEC strains. Methods and Findings Recombinant scFv were developed against ETEC heat-labile toxin (LT) and heat-stable toxin (ST), from previously isolated hybridoma clones. This work reports their design, construction, molecular and functional characterization against LT and ST toxins. Both antibody fragments were able to recognize the cell-interacting toxins by immunofluorescence, the purified toxins by ELISA and also LT-, ST- and LT/ST-producing ETEC strains. Conclusion The developed recombinant scFvs against LT and ST constitute promising starting point for simple and cost-effective ETEC diagnosis. PMID:26154103

  13. Distinct Cytokine Regulation by Cholera Toxin and Type II Heat-Labile Toxins Involves Differential Regulation of CD40 Ligand on CD4+ T Cells

    PubMed Central

    Martin, Michael; Metzger, Daniel J.; Michalek, Suzanne M.; Connell, Terry D.; Russell, Michael W.

    2001-01-01

    Cholera toxin (CT) and the type II heat-labile enterotoxins (HLT) LT-IIa and LT-IIb act as potent systemic and mucosal adjuvants and induce distinct T-helper (Th)-cell cytokine profiles. In the present study, CT and the type II HLT were found to differentially affect cytokine production by anti-CD3-stimulated human peripheral blood mononuclear cells (PBMC), and the cellular mechanisms responsible were investigated. CT suppressed interleukin-2 (IL-2), tumor necrosis factor alpha (TNF-α), and IL-12 production by PBMC cultures more than either LT-IIa or LT-IIb. CT but not LT-IIa or LT-IIb reduced the expression of CD4+ T-cell surface activation markers (CD25 and CD69) and subsequent proliferative responses of anti-CD3-stimulated T cells. CT but not LT-IIa or LT-IIb significantly reduced the expression of CD40 ligand (CD40L) on CD4+ T cells. In a coculture system, CT-treated CD4+ T cells induced significantly less TNF-α and IL-12 p70 production by both autologous monocytes and monocyte-derived dendritic cells than either LT-IIa- or LT-IIb-treated CD4+ T cells. These findings demonstrate that CT, LT-IIa, and LT-IIb differentially affect CD40-CD40L interactions between antigen-presenting cells and T cells and help explain the distinct cytokine profiles observed with type I and type II HLT when used as mucosal adjuvants. PMID:11401990

  14. Effect of heat stable and heat labile Escherichia coli enterotoxins and cholera toxin in combination with theophylline on unidirectional sodium and chloride flux in the small intestine of weanling swine.

    PubMed Central

    Hamilton, D L; Forsyth, G W; Roe, W E; Nielsen, N O

    1978-01-01

    The effect of heat stable and heat labile Escherichia coli enterotoxins or cholera toxin in combination with theophylline on net water, sodium and chloride and unidirectional sodium and chloride fluxes was examined in acute isolated loops of jejunum of weanling swine. The effect of heat stable enterotoxin in combination with theophylline was determined in loops located in the proximal jejunum, while combinations of theophylline and either heat labile enterotoxin or cholera toxin were studied in the distal jejunum. In each situation the addition of theophylline resulted in an additive rather than a synergistic increment of intestinal secretory activity. This study implies that the intestinal adenyl cyclase system and enterotoxin induced intestinal secretion may not be directly related in the swine small intestine. PMID:356942

  15. Effect of Heat Stable and Heat Labile Escherichia coli Enterotoxins, Cholera Toxin and Theophylline on Unidirectional Sodium and Chloride Fluxes in the Proximal and Distal Jejunum of Weanling Swine

    PubMed Central

    Hamilton, D. L.; Roe, W. E.; Nielsen, N. O.

    1977-01-01

    Acute, isolated loops of proximal and distal jejunum of weanling swine were exposed to either heat stable porcine Escherichia coli enterotoxin, heat labile porcine Escherichia coli enterotoxin, cholera toxin or theophylline. Unidirectional sodium fluxes in response to heat stable in the proximal jejunum were dependent on the length of time that the intestinal mucosae was exposed to the enterotoxin. Net water, sodium and chloride and unidirectional sodium and chloride flux measurements in the proximal jejunum in response to each agent uniformly indicated that net secretion of fluid and electrolytes was the result of increased unidirectional sodium secretion or blood-to-lumen flux and decreased unidirectional chloride absorption or lumen-to-blood flux. In addition heat stable cholera toxin and theophylline but not heat labile decreased unidirectional chloride secretion a small but significant amount in the proximal jejunum. Sodium and chloride flux measurements in the distal jejunum demonstrated that all four secretory agents could stimulate net secretion of water, sodium and chloride in that region. The response to these secretory agents as measured by sodium and chloride unidirectional flux rates was not similar to changes observed in the proximal jejunum. In the distal small intestine, whereas heat labile cholera toxin and theophylline induced similar qualitative changes in unidirectional sodium and chloride fluxes, that induced by heat stable differed. PMID:332292

  16. Activation of cholera toxin and Escherichia coli heat-labile enterotoxins by ADP-ribosylation factors, a family of 20 kDa guanine nucleotide-binding proteins.

    PubMed

    Moss, J; Vaughan, M

    1991-11-01

    Cholera toxin and Escherichia coli heat-labile enterotoxins are responsible, in part, for the symptomatology of cholera and traveller's diarrhoea, respectively. Effects of the toxins result from ADP-ribosylation of regulatory guanine nucleotide-binding (G) proteins; the ADP-ribosylated G protein is stabilized in an activated state, resulting in prolonged effects on its target. Toxin-catalysed ADP-ribosylation is stimulated in vitro by a family of guanine nucleotide-binding proteins, c. 20 kDa, termed ADP-ribosylation factors or ARFs. In the presence of GTP, but not GDP or adenine analogues, ARFs serve as allosteric activators of the toxin. The effects are amplified by certain phospholipids and detergents which promote guanine nucleotide binding. Six different mammalian ARF genes have been identified. They encode highly conserved, ubiquitous proteins of 175 to 181 amino acids, containing consensus domains responsible for guanine nucleotide binding. Differences in amino acid sequences are localized near the amino terminus and in the carboxy half of the protein. Although the physiological functions of ARFs have not been precisely defined, their immunological localization to the Golgi is consistent with a role in the regulated orderly movement of newly synthesized proteins from the endoplasmic reticulum, through the Golgi system to their ultimate destination. PMID:1779753

  17. Selection of Escherichia coli heat-labile toxin (LT) inhibitors using both the GM1-ELISA and the cAMP Vero cell assay.

    PubMed

    Verhelst, Roderick; Schroyen, Martine; Buys, Nadine; Niewold, Theo

    2013-07-01

    Weaned piglets are very susceptible to diarrhea caused by enterotoxigenic Escherichia coli. In the past, various natural components were proposed to have beneficial effects by reducing the effects of diarrheal infectious diseases in humans and animals, and thus may represent an alternative for the use of (prophylactic) antibiotics. Alternatives may inactivate enterotoxigenic Escherichia coli heat-labile toxin (LT) by interfering with toxin binding to the cellular receptor GM1. In this study, various plants and other natural substances were tested for inhibitory properties, in the GM1 binding assay, and in the LT-induced cAMP production in Vero cells. The toxic dose of each compound was determined in a cell viability assay, and the highest nontoxic concentrations were used in the GM1 and cAMP assays. Results demonstrated that only d-(+)-galactose, lactose, N-acetyl-d-galactosamine, and two tea extracts were able to inhibit the binding of LT to its GM1 receptor. In the cAMP assay, only the two tea extracts showed inhibitory activity. This shows that d-(+)-galactose, lactose, and N-acetyl-d-galactosamine can indeed inhibit LT binding to GM1 based on structural homology with GM1 in the absence of living cells. However, in the cAMP assay, d-(+)-galactose, and lactose, N-acetyl-d-galactosamine are apparently metabolized to below their effective inhibitory concentration, likely predicting limited practical applicability in vivo. Both tea extracts maintained their activity in the presence of cells. The active compounds in both are probably polyphenols, which are not easily metabolized, and most likely work by aggregating the toxin. In conclusion, the combination of methods used here is a convenient and fast method for preselecting natural substances containing potentially toxin-binding compounds. Furthermore, if antidiarrhea activity is attributed to compounds found inactive here, their activity is unlikely based on interference with toxin binding. PMID:23692076

  18. Escherichia coli Heat-Labile Enterotoxin B Subunit Is a More Potent Mucosal Adjuvant than Its Closely Related Homologue, the B Subunit of Cholera Toxin

    PubMed Central

    Millar, Douglas G.; Hirst, Timothy R.; Snider, Denis P.

    2001-01-01

    Although cholera toxin (Ctx) and Escherichia coli heat-labile enterotoxin (Etx) are known to be potent mucosal adjuvants, it remains controversial whether the adjuvanticity of the holotoxins extends to their nontoxic, receptor-binding B subunits. Here, we have systematically evaluated the comparative adjuvant properties of highly purified recombinant EtxB and CtxB. EtxB was found to be a more potent adjuvant than CtxB, stimulating responses to hen egg lysozyme when the two were coadministered to mice intranasally, as assessed by enhanced serum and secretory antibody titers as well as by stimulation of lymphocyte proliferation in spleen and draining lymph nodes. These results indicate that, although structurally very similar, EtxB and CtxB have strikingly different immunostimulatory properties and should not be considered equivalent as prospective vaccine adjuvants. PMID:11292779

  19. Heterogenic virulence in a diarrheagenic Escherichia coli: evidence for an EPEC expressing heat-labile toxin of ETEC.

    PubMed

    Dutta, Sanjucta; Pazhani, Gururaja P; Nataro, James P; Ramamurthy, Thandavarayan

    2015-01-01

    We have encountered an Escherichia coli strain isolated from a child with acute diarrhea. This strain harbored eae and elt genes encoding for E. coli attaching and effacing property and heat-labile enterotoxin of EPEC and ETEC, respectively. Due to the presence of these distinct virulence factors, we named this uncommon strain as EPEC/ETEC hybrid. The elt gene was identified in a conjugally transferable plasmid of the EPEC/ETEC hybrid. In addition, several virulence genes in the locus of enterocyte effacement have been identified, which confirms that the EPEC/ETEC has an EPEC genetic background. The hybrid nature of this strain was further confirmed by using tissue culture assays. In the multi locus sequence typing (MLST) analysis, the EPEC/ETEC belonged to the sequence type ST328 and was belonging to ST278 Cplx. Sequence analysis of the plasmid DNA revealed presence of six large contigs with several insertion sequences. A phage integrase gene and the prophages of gp48 and gp49 have been found in the upstream of eltAB. In the downstream of elt, an urovirulence loci adhesion encoding (pap) cluster containing papG, and papC were also identified. Similar to other reports, we have identified a heterogenic virulence in a diarrheagenic E. coli but with different combination of genes. PMID:25465159

  20. Structural basis for the differential toxicity of cholera toxin and Escherichia coli heat-labile enterotoxin. Construction of hybrid toxins identifies the A2-domain as the determinant of differential toxicity.

    PubMed

    Rodighiero, C; Aman, A T; Kenny, M J; Moss, J; Lencer, W I; Hirst, T R

    1999-02-12

    Cholera toxin (Ctx) and E. coli heat-labile enterotoxin (Etx) are structurally and functionally similar AB5 toxins with over 80% sequence identity. When their action in polarized human epithelial (T84) cells was monitored by measuring toxin-induced Cl- ion secretion, Ctx was found to be the more potent of the two toxins. Here, we examine the structural basis for this difference in toxicity by engineering a set of mutant and hybrid toxins and testing their activity in T84 cells. This revealed that the differential toxicity of Ctx and Etx was (i) not due to differences in the A-subunit's C-terminal KDEL targeting motif (which is RDEL in Etx), as a KDEL to RDEL substitution had no effect on cholera toxin activity; (ii) not attributable to the enzymatically active A1-fragment, as hybrid toxins in which the A1-fragment in Ctx was substituted for that of Etx (and vice versa) did not alter relative toxicity; and (iii) not due to the B-subunit, as the replacement of the B-subunit in Ctx for that of Etx caused no alteration in toxicity, thus excluding the possibility that the broader receptor specificity of EtxB is responsible for reduced activity. Remarkably, the difference in toxicity could be mapped to a 10-amino acid segment of the A2-fragment that penetrates the central pore of the B-subunit pentamer. A comparison of the in vitro stability of two hybrid toxins, differing only in this 10-amino acid segment, revealed that the Ctx A2-segment conferred a greater stability to the interaction between the A- and B-subunits than the corresponding segment from Etx A2. This suggests that the reason for the relative potency of Ctx compared with Etx stems from the increased ability of the A2-fragment of Ctx to maintain holotoxin stability during uptake and transport into intestinal epithelia. PMID:9933586

  1. Safety and Immunogenicity of an Enterotoxigenic Escherichia coli Vaccine Patch Containing Heat-Labile Toxin: Use of Skin Pretreatment To Disrupt the Stratum Corneum▿

    PubMed Central

    Glenn, Gregory M.; Villar, Christina P.; Flyer, David C.; Bourgeois, A. Louis; McKenzie, Robin; Lavker, Robert M.; Frech, Sarah A.

    2007-01-01

    Transcutaneous immunization allows safe delivery of native heat-labile enterotoxin (LT) from Escherichia coli via application of a simple patch. Physical disruption of the stratum corneum can improve the efficiency of delivery. In the current study, the stratum corneum was disrupted using an electrocardiogram prep pad prior to patch application. The effects were quantified using transepidermal water loss (TEWL) and were correlated with the immune responses. Sixty adults received 50 μg of LT from three lots of LT (20 adults per group) administered in a patch on days 0 and 21. The immunizations were well tolerated. There were no differences in the anti-LT immunoglobulin G (IgG) titers between the three LT lots; the seroconversion rate was 100%, and the mean anti-LT IgG titer was 12,185 enzyme-linked immunosorbent assay units (EU) (a 24-fold increase). TEWL measurements obtained at the time of the second immunization were found to correlate with the day 42 individual increases in the anti-LT IgG titer (r = 0.59, P < 0.001). In a comparative assessment of the immune responses, sera after an LT+ ST+ (E2447A) oral ETEC challenge, which induced moderate to severe diarrhea in 81% of the recipients, had anti-LT IgG titers of 3,245 EU (a 10.8-fold increase). Similarly, the anti-LT IgG titer after administration of an oral cholera toxin B subunit-containing cholera vaccine, which cross-reacts with LT and protects against LT and LT/heat-stable toxin ETEC disease in the field, was 6,741 EU (a 3.3-fold increase). This study confirmed that a well-tolerated regimen for stratum corneum disruption before vaccine patch application results in robust immunity comparable to natural immunity and vaccine-induced immunity and that the magnitude of stratum corneum disruption correlates with the immune response. PMID:17261601

  2. Intranasal immunization with live recombinant Lactococcus lactis combined with heat-labile toxin B subunit protects chickens from highly pathogenic avian influenza H5N1 virus.

    PubMed

    Lei, Han; Peng, Xiaojue; Shu, Handing; Zhao, Daxian

    2015-01-01

    Development of safe and effective vaccines to prevent highly pathogenic avian influenza H5N1 virus infection is a challenging goal. Lactococcus lactis (L. lactis) is an ideal delivery vector for vaccine development, and it has been shown previously that oral immunization of encapsulated secretory L. lactis-hemagglutinin (HA) could provide complete protection against homologous H5N1 virus challenge in the mice model. While intranasal immunization is an appealing approach, it is now reported that secretory L. lactis-HA combined with mucosal adjuvant heat-labile toxin B subunit (LTB) could provide protective immunity in the chicken model. As compared to intranasal immunization with L. lactis-HA alone, L. lactis-HA combined with LTB (L. lactis-HA?+?LTB) could elicit robust neutralizing antibody responses and mucosal IgA responses, as well as strong cellular immune responses in the vaccinated chickens. Importantly, intranasal immunization with L. lactis-HA?+?LTB could provide 100% protection against H5N1 virus challenge. Taken together, these results suggest that intranasal immunization with L. lactis-HA?+?LTB can be considered as an effective approach for preventing and controlling infection of H5N1 virus in poultry during an avian influenza A/H5N1 pandemic. PMID:24861477

  3. Non-recombinant display of the B subunit of the heat labile toxin of Escherichia coli on wild type and mutant spores of Bacillus subtilis

    PubMed Central

    2013-01-01

    Background Mucosal infections are a major global health problem and it is generally accepted that mucosal vaccination strategies, able to block infection at their entry site, would be preferable with respect to other prevention approaches. However, there are still relatively few mucosal vaccines available, mainly because of the lack of efficient delivery systems and of mucosal adjuvants. Recombinant bacterial spores displaying a heterologous antigen have been shown to induce protective immune responses and, therefore, proposed as a mucosal delivery system. A non-recombinant approach has been recently developed and tested to display antigens and enzymes. Results We report that the binding subunit of the heat-labile toxin (LTB) of Escherichia coli efficiently adsorbed on the surface of Bacillus subtilis spores. When nasally administered to groups of mice, spore-adsorbed LTB was able to induce a specific immune response with the production of serum IgG, fecal sIgA and of IFN-? in spleen and mesenteric lymph nodes (MLN) of the immunized animals. Dot blotting experiments showed that the non-recombinant approach was more efficient than the recombinant system in displaying LTB and that the efficiency of display could be further increased by using mutant spores with an altered surface. In addition, immunofluorescence microscopy experiments showed that only when displayed on the spore surface by the non-recombinant approach LTB was found in its native, pentameric form. Conclusion Our results indicate that non-recombinant spores displaying LTB pentamers can be administered by the nasal route to induce a Th1-biased, specific immune response. Mutant spores with an altered coat are more efficient than wild type spores in adsorbing the antigen, allowing the use of a reduced number of spores in immunization procedures. Efficiency of display, ability to display the native form of the antigen and to induce a specific immune response propose this non-recombinant delivery system as a powerful mucosal vaccine delivery approach. PMID:24168229

  4. The catalytic A1 domains of cholera toxin and heat-labile enterotoxin are potent DNA adjuvants that evoke mixed Th1/Th17 cellular immune responses

    PubMed Central

    Bagley, Kenneth; Xu, Rong; Ota-Setlik, Ayuko; Egan, Michael; Schwartz, Jennifer; Fouts, Timothy

    2015-01-01

    DNA encoded adjuvants are well known for increasing the magnitude of cellular and/or humoral immune responses directed against vaccine antigens. DNA adjuvants can also tune immune responses directed against vaccine antigens to better protect against infection of the target organism. Two potent DNA adjuvants that have unique abilities to tune immune responses are the catalytic A1 domains of Cholera Toxin (CTA1) and Heat-Labile Enterotoxin (LTA1). Here, we have characterized the adjuvant activities of CTA1 and LTA1 using HIV and SIV genes as model antigens. Both of these adjuvants enhanced the magnitude of antigen-specific cellular immune responses on par with those induced by the well-characterized cytokine adjuvants IL-12 and GM-CSF. CTA1 and LTA1 preferentially enhanced cellular responses to the intracellular antigen SIVmac239-gag over those for the secreted HIVBaL-gp120 antigen. IL-12, GM-CSF and electroporation did the opposite suggesting differences in the mechanisms of actions of these diverse adjuvants. Combinations of CTA1 or LTA1 with IL-12 or GM-CSF generated additive and better balanced cellular responses to both of these antigens. Consistent with observations made with the holotoxin and the CTA1-DD adjuvant, CTA1 and LTA1 evoked mixed Th1/Th17 cellular immune responses. Together, these results show that CTA1 and LTA1 are potent DNA vaccine adjuvants that favor the intracellular antigen gag over the secreted antigen gp120 and evoke mixed Th1/Th17 responses against both of these antigens. The results also indicate that achieving a balanced immune response to multiple intracellular and extracellular antigens delivered via DNA vaccination may require combining adjuvants that have different and complementary mechanisms of action. PMID:26042527

  5. Parenteral Adjuvant Activities of Escherichia coli Heat-Labile Toxin and Its B Subunit for Immunization of Mice against Gastric Helicobacter pylori Infection

    PubMed Central

    Weltzin, Richard; Guy, Bruno; Thomas, William D.; Giannasca, Paul J.; Monath, Thomas P.

    2000-01-01

    The heat-labile toxin (LT) of Escherichia coli is a potent mucosal adjuvant that has been used to induce protective immunity against Helicobacter felis and Helicobacter pylori infection in mice. We studied whether recombinant LT or its B subunit (LTB) has adjuvant activity in mice when delivered with H. pylori urease antigen via the parenteral route. Mice were immunized subcutaneously or intradermally with urease plus LT, recombinant LTB, or a combination of LT and LTB prior to intragastric challenge with H. pylori. Control mice were immunized orally with urease plus LT, a regimen shown previously to protect against H. pylori gastric infection. Parenteral immunization using either LT or LTB as adjuvant protected mice against H. pylori challenge as effectively as oral immunization and enhanced urease-specific immunoglobulin G (IgG) responses in serum as effectively as aluminum hydroxide adjuvant. LT and LTB had adjuvant activity at subtoxic doses and induced more consistent antibody responses than those observed with oral immunization. A mixture of a low dose of LT and a high dose of LTB stimulated the highest levels of protection and specific IgG in serum. Urease-specific IgG1 and IgG2a antibody subclass responses were stimulated by all immunization regimens tested, but relative levels were dependent on the adjuvant used. Compared to parenteral immunization with urease alone, LT preferentially enhanced IgG1, while LTB or the LT-LTB mixture preferentially enhanced IgG2a. Parenteral immunization using LT or LTB as adjuvant also induced IgA to urease in the saliva of some mice. These results show that LT and LTB stimulate qualitatively different humoral immune responses to urease but are both effective parenteral adjuvants for immunization of mice against H. pylori infection. PMID:10768972

  6. Evaluating the A-Subunit of the Heat-Labile Toxin (LT) As an Immunogen and a Protective Antigen Against Enterotoxigenic Escherichia coli (ETEC).

    PubMed

    Norton, Elizabeth B; Branco, Luis M; Clements, John D

    2015-01-01

    Diarrheal illness contributes to malnutrition, stunted growth, impaired cognitive development, and high morbidity rates in children worldwide. Enterotoxigenic Escherichia coli (ETEC) is a major contributor to this diarrheal disease burden. ETEC cause disease in the small intestine by means of colonization factors and by production of a heat-labile enterotoxin (LT) and/or a small non-immunogenic heat-stable enterotoxin (ST). Overall, the majority of ETEC produce both ST and LT. LT induces secretion via an enzymatically active A-subunit (LT-A) and a pentameric, cell-binding B-subunit (LT-B). The importance of anti-LT antibodies has been demonstrated in multiple clinical and epidemiological studies, and a number of potential ETEC vaccine candidates have included LT-B as an important immunogen. However, there is limited information about the potential contribution of LT-A to development of protective immunity. In the current study, we evaluate the immune response against the A-subunit of LT as well as the A-subunit's potential as a protective antigen when administered alone or in combination with the B-subunit of LT. We evaluated human sera from individuals challenged with a prototypic wild-type ETEC strain as well as sera from individuals living in an ETEC endemic area for the presence of anti-LT, anti-LT-A and anti-LT-B antibodies. In both cases, a significant number of individuals intentionally or endemically infected with ETEC developed antibodies against both LT subunits. In addition, animals immunized with the recombinant proteins developed robust antibody responses that were able to neutralize the enterotoxic and cytotoxic effects of native LT by blocking binding and entry into cells (anti-LT-B) or the intracellular enzymatic activity of the toxin (anti-LT-A). Moreover, antibodies to both LT subunits acted synergistically to neutralize the holotoxin when combined. Taken together, these data support the inclusion of both LT-A and LT-B in prospective vaccines against ETEC. PMID:26305793

  7. Evaluating the A-Subunit of the Heat-Labile Toxin (LT) As an Immunogen and a Protective Antigen Against Enterotoxigenic Escherichia coli (ETEC)

    PubMed Central

    Norton, Elizabeth B.; Branco, Luis M.; Clements, John D.

    2015-01-01

    Diarrheal illness contributes to malnutrition, stunted growth, impaired cognitive development, and high morbidity rates in children worldwide. Enterotoxigenic Escherichia coli (ETEC) is a major contributor to this diarrheal disease burden. ETEC cause disease in the small intestine by means of colonization factors and by production of a heat-labile enterotoxin (LT) and/or a small non-immunogenic heat-stable enterotoxin (ST). Overall, the majority of ETEC produce both ST and LT. LT induces secretion via an enzymatically active A-subunit (LT-A) and a pentameric, cell-binding B-subunit (LT-B). The importance of anti-LT antibodies has been demonstrated in multiple clinical and epidemiological studies, and a number of potential ETEC vaccine candidates have included LT-B as an important immunogen. However, there is limited information about the potential contribution of LT-A to development of protective immunity. In the current study, we evaluate the immune response against the A-subunit of LT as well as the A-subunit’s potential as a protective antigen when administered alone or in combination with the B-subunit of LT. We evaluated human sera from individuals challenged with a prototypic wild-type ETEC strain as well as sera from individuals living in an ETEC endemic area for the presence of anti-LT, anti-LT-A and anti-LT-B antibodies. In both cases, a significant number of individuals intentionally or endemically infected with ETEC developed antibodies against both LT subunits. In addition, animals immunized with the recombinant proteins developed robust antibody responses that were able to neutralize the enterotoxic and cytotoxic effects of native LT by blocking binding and entry into cells (anti-LT-B) or the intracellular enzymatic activity of the toxin (anti-LT-A). Moreover, antibodies to both LT subunits acted synergistically to neutralize the holotoxin when combined. Taken together, these data support the inclusion of both LT-A and LT-B in prospective vaccines against ETEC. PMID:26305793

  8. Heat-Labile Enterotoxin: Beyond GM1 Binding

    PubMed Central

    Mudrak, Benjamin; Kuehn, Meta J.

    2010-01-01

    Enterotoxigenic Escherichia coli (ETEC) is a significant source of morbidity and mortality worldwide. One major virulence factor released by ETEC is the heat-labile enterotoxin LT, which is structurally and functionally similar to cholera toxin. LT consists of five B subunits carrying a single catalytically active A subunit. LTB binds the monosialoganglioside GM1, the toxins host receptor, but interactions with A-type blood sugars and E. coli lipopolysaccharide have also been identified within the past decade. Here, we review the regulation, assembly, and binding properties of the LT B-subunit pentamer and discuss the possible roles of its numerous molecular interactions. PMID:22069646

  9. Current progress in the development of the B subunits of cholera toxin and Escherichia coli heat-labile enterotoxin as carriers for the oral delivery of heterologous antigens and epitopes.

    PubMed

    Nashar, T O; Amin, T; Marcello, A; Hirst, T R

    1993-01-01

    The development of non-living carrier systems for delivery of protective antigens or epitopes to the immune system represents both a fundamental and an applied aspect of vaccinology. A wide range of carrier systems, ranging from inert supports to proteins that exert direct immunomodulating effects on the immune response, are being studied. In this overview we describe the current progress in the development of the B-subunits of cholera toxin and Escherichia coli heat-labile enterotoxin as potential protein carriers for the oral delivery of chemically and genetically attached antigens and epitopes. PMID:7679865

  10. Rapid differentiation of enterotoxigenic Escherichia coli that produce heat-stable and heat-labile toxins by frequency-pulsed electron capture gas-liquid chromatography analysis of diarrheal stool specimens.

    PubMed Central

    Brooks, J B; Basta, M T; el Kholy, A M; Moss, C W

    1984-01-01

    Thirty-three stool specimens from infants in the village of Tamooh near Cairo, Egypt, were studied by frequency-pulsed electron capture gas-liquid chromatography (FPEC-GLC). In 13 of the diarrheal cases, the suspected causative agent isolated was Escherichia coli which produced heat-stable toxin (ST), and in 10 other cases E. coli that produced heat-labile toxin (LT) were isolated. Ten control stool samples, collected from infants from whom no pathogenic organisms were isolated, were analyzed at the same time. Comparisons also were made against healthy control stools from individuals in the United States who had been previously analyzed by FPEC-GLC (Brooks et al., J. Clin. Microbiol. 20:549-560, 1984). The stools were suspended in water and centrifuged, and the supernatant was extracted with organic solvents and derivatized to form electron-capturing derivatives of carboxylic acids, hydroxy acids, alcohols, and amines. Results from the study showed distinct differences among the FPEC-GLC profiles of E. coli ST-positive stools, of E. coli LT-positive stools, and of the control stool samples. An unidentified compound appearing in the ether-soluble hydroxy acid fraction from E. coli ST-positive stools was tentatively identified by mass spectrometry as 6-methoxy-2-hydroxyhexanoic acid. 6-Methoxy-2-hydroxyhexanoic acid was found in all stools that contained E. coli ST but was not present either in stools from which E. coli LT was isolated or in control samples. 6-Methoxy-2-hydroxyhexanoic acid may prove to be an important marker for use in the identification of E. coli ST. In addition to 6-methoxy-2-hydroxyhexanoic acid, the carboxylic acid, alcohol, and amine FPEC-GLC profiles obtained from stools were very different between these two organisms. The data indicate that FPEC-GLC analysis of diarrheal stool specimens might be a rapid way to distinguish diarrhea caused by E. coli ST, E. coli LT, Clostridium difficile, and rotavirus. PMID:6394617

  11. Rapid differentiation of enterotoxigenic Escherichia coli that produce heat-stable and heat-labile toxins by frequency-pulsed electron capture gas-liquid chromatography analysis of diarrheal stool specimens.

    PubMed

    Brooks, J B; Basta, M T; el Kholy, A M; Moss, C W

    1984-12-01

    Thirty-three stool specimens from infants in the village of Tamooh near Cairo, Egypt, were studied by frequency-pulsed electron capture gas-liquid chromatography (FPEC-GLC). In 13 of the diarrheal cases, the suspected causative agent isolated was Escherichia coli which produced heat-stable toxin (ST), and in 10 other cases E. coli that produced heat-labile toxin (LT) were isolated. Ten control stool samples, collected from infants from whom no pathogenic organisms were isolated, were analyzed at the same time. Comparisons also were made against healthy control stools from individuals in the United States who had been previously analyzed by FPEC-GLC (Brooks et al., J. Clin. Microbiol. 20:549-560, 1984). The stools were suspended in water and centrifuged, and the supernatant was extracted with organic solvents and derivatized to form electron-capturing derivatives of carboxylic acids, hydroxy acids, alcohols, and amines. Results from the study showed distinct differences among the FPEC-GLC profiles of E. coli ST-positive stools, of E. coli LT-positive stools, and of the control stool samples. An unidentified compound appearing in the ether-soluble hydroxy acid fraction from E. coli ST-positive stools was tentatively identified by mass spectrometry as 6-methoxy-2-hydroxyhexanoic acid. 6-Methoxy-2-hydroxyhexanoic acid was found in all stools that contained E. coli ST but was not present either in stools from which E. coli LT was isolated or in control samples. 6-Methoxy-2-hydroxyhexanoic acid may prove to be an important marker for use in the identification of E. coli ST. In addition to 6-methoxy-2-hydroxyhexanoic acid, the carboxylic acid, alcohol, and amine FPEC-GLC profiles obtained from stools were very different between these two organisms. The data indicate that FPEC-GLC analysis of diarrheal stool specimens might be a rapid way to distinguish diarrhea caused by E. coli ST, E. coli LT, Clostridium difficile, and rotavirus. PMID:6394617

  12. Ability of SPI2 mutant of S. typhi to effectively induce antibody responses to the mucosal antigen enterotoxigenic E. coli heat labile toxin B subunit after oral delivery to humans

    PubMed Central

    Khan, S.; Chatfield, S.; Stratford, R.; Bedwell, J.; Bentley, M.; Sulsh, S.; Giemza, R.; Smith, S.; Bongard, E.; Cosgrove, C.A.; Johnson, J.; Dougan, G.; Griffin, G.E.; Makin, J.; Lewis, D.J.M.

    2007-01-01

    We have evaluated an oral vaccine based on an Salmonella enteric serovar typhi (S. typhi) Ty2 derivative TSB7 harboring deletion mutations in ssaV (SPI-2) and aroC together with a chromosomally integrated copy of eltB encoding the B subunit of enterotoxigenic Escherichia coli heat labile toxin (LT-B) in volunteers. Two oral doses of 108 or 109 CFU were administered to two groups of volunteers and both doses were well tolerated, with no vaccinemia, and only transient stool shedding. Immune responses to LT-B and S. typhi lipopolysaccharide were demonstrated in 67 and 97% of subjects, respectively, without evidence of anti-carrier immunity preventing boosting of LT-B responses in many cases. Further development of this salmonella-based (spi-VEC) system for oral delivery of heterologous antigens appears warranted. PMID:17412462

  13. Cholera toxin, LT-I, LT-IIa, and LT-IIb: the critical role of ganglioside-binding in immunomodulation by Type I and Type II heat-labile enterotoxins

    PubMed Central

    Connell, Terry D.

    2010-01-01

    The heat-labile enterotoxins (HLT) expressed by Vibrio cholerae (cholera toxin) and Escherichia coli (LT-I, LT-IIa, and LT-IIb) are potent systemic and mucosal adjuvants. Co-administration of the enterotoxins with a foreign antigen (Ag) produces an augmented immune response to that antigen. Although each enterotoxin has potent adjuvant properties, the means by which the enterotoxins induce various immune responses are distinctive for each adjuvant. Various mutants have been engineered to dissect the functions of the enterotoxins required for their adjuvanticity. The capacity to strongly bind to one or more specific ganglioside receptors appears to drive the distinctive immunomodulatory properties associated with each enterotoxin. Mutant enterotoxins with ablated or altered ganglioside binding affinities have been employed to investigate the role of gangliosides in enterotoxin-dependent immunomodulation. PMID:17931161

  14. Protective Effect of Active Immunization with Purified Escherichia coli Heat-Labile Enterotoxin in Rats

    PubMed Central

    Klipstein, Frederick A.; Engert, Richard F.

    1979-01-01

    The protective effect of active immunization by different routes with a purified preparation of the polymyxin-release form of Escherichia coli heat-labile toxin was evaluated in rats. Immunized animals were challenged by placing toxin into ligated ileal loops at dosages which produced either 50% or the maximum secretory response in unimmunized rats. Immunization exclusively by the parenteral route yielded significant protection. Rats were also protected when parenteral priming was followed by boosting given either directly into the duodenum or perorally 2 h after intragastric cimetidine, but not when the peroral boosts were given with bicarbonate. Immunization administered entirely by the peroral route with cimetidine yielded protection but only when the immunizing dosage was fivefold greater than that found effective in the parenteral-peroral approach. Rats immunized exclusively by the parenteral route and those boosted perorally with cimetidine were also tested, and found to be protected, against challenge with viable organisms of strains that produce either heat-labile toxin alone or both heat-labile and heat-stable toxin, but they were not protected against a strain which produces just heat-stable toxin. Geometric mean serum antibody titers were increased by 16-fold or more over control values in those groups of rats in which protection was achieved, with the exception of those immunized exclusively by the peroral route. These observations demonstrate that (i) active immunization with purified E. coli heat-labile toxin results in significant protection against both this toxin as well as viable organisms which produce it, but not against viable strains which produce heat-stable toxin only, and (ii) concomitant ablation of gastric secretion by the use of cimetidine renders the peroral route of immunization effective. They suggest that prophylactic immunization against diarrheal disease caused by heat-labile toxin-producing strains of E. coli may be feasible in humans. PMID:378831

  15. The B subunit of Escherichia coli heat-labile toxin alters the development and antigen-presenting capacity of dendritic cells

    PubMed Central

    Ji, Jing; Griffiths, Kristin L; Milburn, Peter J; Hirst, Timothy R; O’Neill, Helen C

    2015-01-01

    Escherichia coli’s heat-labile enterotoxin (Etx) and its non-toxic B subunit (EtxB) have been characterized as adjuvants capable of enhancing T cell responses to co-administered antigen. Here, we investigate the direct effect of intravenously administered EtxB on the size of the dendritic and myeloid cell populations in spleen. EtxB treatment appears to enhance the development and turnover of dendritic and myeloid cells from precursors within the spleen. EtxB treatment also gives a dendritic cell (DC) population with higher viability and lower activation status based on the reduced expression of MHC-II, CD80 and CD86. In this respect, the in vivo effect of EtxB differs from that of the highly inflammatory mediator lipopolysaccharide. In in vitro bone marrow cultures, EtxB treatment was also found to enhance the development of DC from precursors dependent on Flt3L. In terms of the in vivo effect of EtxB on CD4 and CD8 T cell responses in mice, the interaction of EtxB directly with DC was demonstrated following conditional depletion of CD11c+ DC. In summary, all results are consistent with EtxB displaying adjuvant ability by enhancing the turnover of DC in spleen, leading to newly mature myeloid and DC in spleen, thereby increasing DC capacity to perform as antigen-presenting cells on encounter with T cells. PMID:26130503

  16. Gangliosides sensitize unresponsive fibroblasts to Escherichia coli heat-labile enterotoxin.

    PubMed

    Moss, J; Garrison, S; Fishman, P H; Richardson, S H

    1979-08-01

    Chemically transformed mouse fibroblasts did not raise their cyclic AMP level in response to Escherichia coli heat-labile enterotoxin. These fibroblasts did, however, incorporate exogenous mono-, di-, and trisialogangliosides. After the uptake of monosialoganglioside galactosyl-N-acetylgalactosaminyl-[N-acetylneuraminyl]-galactosylglucosylceramide (GM1), the cells responded to E. coli heat-labile enterotoxin. The di- and trisialogangliosides were considerably less effective. GM1, the putative cholera toxin (choleragen) receptor, has been implicated previously as the receptor for E. coli heat-labile enterotoxin based on the ability of the free ganglioside to inhibit the effects of toxin. This investigation establishes that the ganglioside, when incorporated into fibroblasts, serves a functional role in mediating the responsiveness to the toxin. PMID:222809

  17. Production of type II heat-labile enterotoxin by Escherichia coli isolated from food and human feces.

    PubMed Central

    Guth, B E; Pickett, C L; Twiddy, E M; Holmes, R K; Gomes, T A; Lima, A A; Guerrant, R L; Franco, B D; Trabulsi, L R

    1986-01-01

    Escherichia coli strains isolated in Sao Paulo, Brazil, from feces of patients with diarrhea and from food samples produced toxin(s) that was shown to be related both immunologically and genetically to the recently characterized type II heat-labile enterotoxin of E. coli. The new isolates of type II heat-labile enterotoxin-producing E. coli belonged to five different serotypes and did not represent a single clone. Images PMID:3533784

  18. Genetic fusions of a CFA/I/II/IV MEFA (multiepitope fusion antigen) and a toxoid fusion of heat-stable toxin (STa) and heat-labile toxin (LT) of enterotoxigenic Escherichia coli (ETEC) retain broad anti-CFA and antitoxin antigenicity.

    PubMed

    Ruan, Xiaosai; Sack, David A; Zhang, Weiping

    2015-01-01

    Immunological heterogeneity has long been the major challenge in developing broadly effective vaccines to protect humans and animals against bacterial and viral infections. Enterotoxigenic Escherichia coli (ETEC) strains, the leading bacterial cause of diarrhea in humans, express at least 23 immunologically different colonization factor antigens (CFAs) and two distinct enterotoxins [heat-labile toxin (LT) and heat-stable toxin type Ib (STa or hSTa)]. ETEC strains expressing any one or two CFAs and either toxin cause diarrhea, therefore vaccines inducing broad immunity against a majority of CFAs, if not all, and both toxins are expected to be effective against ETEC. In this study, we applied the multiepitope fusion antigen (MEFA) strategy to construct ETEC antigens and examined antigens for broad anti-CFA and antitoxin immunogenicity. CFA MEFA CFA/I/II/IV [CVI 2014, 21(2):243-9], which carried epitopes of seven CFAs [CFA/I, CFA/II (CS1, CS2, CS3), CFA/IV (CS4, CS5, CS6)] expressed by the most prevalent and virulent ETEC strains, was genetically fused to LT-STa toxoid fusion monomer 3xSTaA14Q-dmLT or 3xSTaN12S-dmLT [IAI 2014, 82(5):1823-32] for CFA/I/II/IV-STaA14Q-dmLT and CFA/I/II/IV-STaN12S-dmLT MEFAs. Mice intraperitoneally immunized with either CFA/I/II/IV-STa-toxoid-dmLT MEFA developed antibodies specific to seven CFAs and both toxins, at levels equivalent or comparable to those induced from co-administration of the CFA/I/II/IV MEFA and toxoid fusion 3xSTaN12S-dmLT. Moreover, induced antibodies showed in vitro adherence inhibition activities against ETEC or E. coli strains expressing these seven CFAs and neutralization activities against both toxins. These results indicated CFA/I/II/IV-STa-toxoid-dmLT MEFA or CFA/I/II/IV MEFA combined with 3xSTaN12S-dmLT induced broadly protective anti-CFA and antitoxin immunity, and suggested their potential application in broadly effective ETEC vaccine development. This MEFA strategy may be generally used in multivalent vaccine development. PMID:25803825

  19. Inhibition of heat-labile cholera and Escherichia coli enterotoxins by brefeldin A.

    PubMed Central

    Donta, S T; Beristain, S; Tomicic, T K

    1993-01-01

    Cholera enterotoxin and the related heat-labile enterotoxins of Escherichia coli enter their target cells through noncoated vesicles, but how the toxins are processed intracellularly and how they get to their targeted enzyme, adenylate cyclase, remain to be defined. Brefeldin A, an inhibitor of the trans-Golgi network, is shown herein to transiently block the morphologic and enzymatic effects of the toxin at a step distal to the initial binding process but prior to activation of adenylate cyclase by the toxin. It is likely, therefore, that these toxins are processed by the Golgi apparatus before trafficking to the membrane adenylate cyclase. Images PMID:8392970

  20. LT-IIb(T13I), a non-toxic type II heat-labile enterotoxin, augments the capacity of a ricin toxin subunit vaccine to evoke neutralizing antibodies and protective immunity.

    PubMed

    Greene, Christopher J; Chadwick, Chrystal M; Mandell, Lorrie M; Hu, John C; O'Hara, Joanne M; Brey, Robert N; Mantis, Nicholas J; Connell, Terry D

    2013-01-01

    Currently, there is a shortage of adjuvants that can be employed with protein subunit vaccines to enhance protection against biological threats. LT-IIb(T13I) is an engineered nontoxic derivative of LT-IIb, a member of the type II subfamily of heat labile enterotoxins expressed by Escherichia coli, that possesses potent mucosal adjuvant properties. In this study we evaluated the capacity of LT-IIb(T13I) to augment the potency of RiVax, a recombinant ricin toxin A subunit vaccine, when co-administered to mice via the intradermal (i.d.) and intranasal (i.n.) routes. We report that co-administration of RiVax with LT-IIb(T13I) by the i.d. route enhanced the levels of RiVax-specific serum IgG antibodies (Ab) and elevated the ratio of ricin-neutralizing to non-neutralizing Ab, as compared to RiVax alone. Protection against a lethal ricin challenge was also augmented by LT-IIb(T13I). While local inflammatory responses elicited by LT-IIb(T13I) were comparable to those elicited by aluminum salts (Imject), LT-IIb(T13I) was more effective than aluminum salts at augmenting production of RiVax-specific serum IgG. Finally, i.n. administration of RiVax with LT-IIb(T13I) also increased levels of RiVax-specific serum and mucosal Ab and enhanced protection against ricin challenge. Collectively, these data highlight the potential of LT-IIb(T13I) as an effective next-generation i.d., or possibly i.n. adjuvant for enhancing the immunogenicity of subunit vaccines for biodefense. PMID:23936344

  1. Immunological cross-reactivity of heat-labile enterotoxins produced by enterotoxigenic and enteropathogenic strains of Escherichia coli.

    PubMed Central

    Klipstein, F A; Engert, R F; Short, H B

    1980-01-01

    Weanling rats were immunized with a heat-labile enterotoxin contained in whole cell lysate (WCL) ultrafiltrate preparations of enteropathogenic (EPEC) and enterotoxigenic (ETEC) strains or with a purified preparation of heat-labile toxin (LT) from the ETEC strain and then challenged either with viable bacteria of each strain or the purified ETEC LT by means of the ileal ligated loop technique. Immunization with the WCL toxin preparations of either the EPEC or ETEC strain conferred protection against challenge with viable organisms of both strains; immunization with a similar preparation from a nontoxigenic strain did not yield protection. Immunization with either the WCL or purified LT toxin from ETEC strain afforded protection against challenge with the ETEC LT toxin, but immunization with the EPEC WCL preparation did not. The antigenicity of all of the toxin preparations was destroyed by heat-treatment. Possible contributory protective effects of somatic or colonization factor (CFA) antigens present in the WCL were excluded by the findings that protection was afforded against a heterologous somatic serotype, ileal bacterial counts were not reduced in protected animals, and WCL preparations of strains containing or lacking CFA yielded equal protection. These observations indicate that the heat-labile enterotoxin of EPEC strains is antigenic and is immunologically related to a heat-labile toxin present in similarly prepared material from an ETEC strain but not to the conventional LT toxin of ETEC strains. They suggest that the WCL preparation of the ETEC strain contains two heat-labile enterotoxins, one of which is conventional LT and the other of which resembles the EPEC toxin. PMID:7000692

  2. Modified Heat-Stable Toxins (hSTa) of Enterotoxigenic Escherichia coli Lose Toxicity but Display Antigenicity after Being Genetically Fused to Heat-Labile Toxoid LT(R192G)

    PubMed Central

    Liu, Mei; Zhang, Chengxian; Mateo, Kristy; Nataro, James P.; Robertson, Donald C.; Zhang, Weiping

    2011-01-01

    Enterotoxigenic Escherichia coli (ETEC) strains are a major cause of diarrhea in humans and animals. Heat-stable (STa) and heat-labile (LT) enterotoxins produced by ETEC disrupt fluid homeostasis in host small intestinal epithelial cells and cause fluid and electrolyte hyper-secretion that leads to diarrhea. ETEC strains producing STa or LT are sufficiently virulent to cause diarrhea, therefore STa and LT antigens must be included in ETEC vaccines. However, potent toxicity and poor immunogenicity (of STa) prevent them from being directly applied as vaccine components. While LT toxoids, especially LT(R192G), being used in vaccine development, STa toxoids have not been included. A recent study (IAI, 78:316-325) demonstrated porcine-type STa toxoids [pSTa(P12F) and pSTa(A13Q)] elicited protective anti-STa antibodies after being fused to a porcine-type LT toxoid [pLT(R192G)]. In this study, we substituted the 8th, 9th, 16th, or the 17th amino acid of a human-type STa (hSTa) and generated 28 modified STa peptides. We tested each STa peptide for toxicity and structure integrity, and found nearly all modified STa proteins showed structure alteration and toxicity reduction. Based on structure similarity and toxic activity, three modified STa peptides: STa(E8A), STa(T16Q) and STa(G17S), were selected to construct LT192-STa-toxoid fusions. Constructed fusions were used to immunize mice, and immunized mice developed anti-STa antibodies. Results from this study provide useful information in developing toxoid vaccines against ETEC diarrhea. PMID:22069760

  3. Analysis and modeling of heat-labile enterotoxins of Escherichia coli suggests a novel space with insights into receptor preference.

    PubMed

    Krishna Raja, M; Ghosh, Asit Ranjan; Vino, S; Sajitha Lulu, S

    2015-01-01

    Features of heat-labile enterotoxins of Escherichia coli which make them fit to use as novel receptors for antidiarrheals are not completely explored. Data-set of 14 different serovars of enterotoxigenic Escherichia coli producing heat-labile toxins were taken from NCBI Genbank database and used in the study. Sequence analysis showed mutations in different subunits and also at their interface residues. As these toxins lack crystallography structures, homology modeling using Modeller 9.11 led to the structural approximation for the E. coli producing heat-labile toxins. Interaction of modeled toxin subunits with proanthocyanidin, an antidiarrheal showed several strong hydrogen bonding interactions at the cost of minimized energy. The hits were subsequently characterized by molecular dynamics simulation studies to monitor their binding stabilities. This study looks into novel space where the ligand can choose the receptor preference not as a whole but as an individual subunit. Mutation at interface residues and interaction among subunits along with the binding of ligand to individual subunits would help to design a non-toxic labile toxin and also to improve the therapeutics. PMID:25375068

  4. Preparation of procoligenoids from Escherichia coli heat-labile enterotoxins.

    PubMed Central

    Finkelstein, R A; Sciortino, C V; Rieke, L C; Burks, M F; Boesman-Finkelstein, M

    1984-01-01

    Heat-labile enterotoxins from Escherichia coli strains of porcine and human origin polymerize on heating to form high-molecular-weight aggregates, "procoligenoids," analogous to procholeragenoid derived from the cholera enterotoxin. This aggregation is accompanied by loss of biological activity (toxicity). Further heating results in the release of B-subunit oligomers, coligenoids, analogous to choleragenoid. Further studies are needed to determine whether, like procholeragenoid, the procoligenoids are superior antigens in stimulating gut immunity after parenteral administration. Images PMID:6378800

  5. Escherichia coli heat-labile toxin subunit B fusions with Streptococcus sobrinus antigens expressed by Salmonella typhimurium oral vaccine strains: importance of the linker for antigenicity and biological activities of the hybrid proteins.

    PubMed Central

    Jagusztyn-Krynicka, E K; Clark-Curtiss, J E; Curtiss, R

    1993-01-01

    A set of vectors possessing the genes for aspartate semialdehyde dehydrogenase (asd) and the B subunit of the heat-labile enterotoxin of Escherichia coli (LT-B) has been developed. These vectors allow operon or gene fusions of foreign gene epitopes at the C-terminal end of LT-B. Two groups of vectors have been constructed with and without leader sequences to facilitate placing of the foreign antigen in different cell compartments. Two Streptococcus sobrinus genes coding for principal colonization factors, surface protein antigen A (SpaA), and dextranase (Dex), have been fused into the 3' end of the LT-B gene. Resulting protein fusions of approximately 120 to 130 kDa are extremely well recognized by antibodies directed against both SpaA and Dex as well as against LT-B domains and retain the enzymatic activity of dextranase and the biological activity of LT-B in that they bind to GM1 gangliosides. Maximum antigenicity was obtained with the vector possessing an intervening linker of at least six amino acids with two proline residues. Some of the fusion proteins also exhibited another property of LT-B in that they were exported into the periplasm where they oligomerized. LT-B-SpaA and LT-B-Dex hybrid proteins are expressed stably and at a high level in avirulent Salmonella typhimurium vaccine strains which are being used to investigate their immunogenicity and types of induced immune responses. The fusion vectors will also be useful for production and purification of LT-B fusion antigens to be used and evaluated in other vaccine compositions. Images PMID:8432584

  6. Heat-Labile Enterotoxin IIa, a Platform To Deliver Heterologous Proteins into Neurons

    PubMed Central

    Chen, Chen; Przedpelski, Amanda; Tepp, William H.; Pellett, Sabine; Johnson, Eric A.

    2015-01-01

    ABSTRACT Cholera toxin (CT) and the related heat-labile enterotoxins (LT) of Escherichia coli have been implicated as adjuvants in human therapies, but reactivity upon intranasal delivery dampened efforts to develop other clinical applications. However, each CT family member variant has unique biological properties that may warrant development as therapeutic platforms. In the current study, a nontoxic variant of the heat-labile enterotoxin IIa (LTIIa) was engineered to deliver heterologous, functional proteins into the cytosol of neurons. As proof of principle, the LTIIa variant delivered two cargos into neurons. LTIIa delivered ?-lactamase efficiently into cells containing complex gangliosides, such as GD1b, as host receptors. LTIIa delivery of ?-lactamase was sensitive to brefeldin A, an inhibitor that collapses the Golgi compartment into the endoplasmic reticulum, but not sensitive to treatment with botulinum neurotoxin D (BoNT/D), an inhibitor of synaptic vesicle cycling. LTIIa delivered a single-chain, anti-BoNT/A camelid antibody that inhibited SNAP25 cleavage during post-BoNT/A exposure of neurons. Delivery of functional, heterologous protein cargos into neurons demonstrates the potential of LTII variants as platforms to deliver therapies to inactivate toxins and microbial infections and to reverse the pathology of human neurodegenerative diseases. PMID:26265718

  7. Effect of fractions of Ethiopian And Norwegian colostrum on rotavirus and Escherichia coli heat-labile enterotoxin.

    PubMed Central

    Otnaess, A B; Orstavik, I

    1981-01-01

    Samples of colostrum from both Ethiopian and Norwegian women contained antirotavirus activities of immunoglobulin and non-immunoglobulin nature. No significant differences in rotavirus immunoglobulin A or in rotavirus-inhibiting activity were found between samples from the two countries. The non-immunoglobulin inhibitory activity was trypsin sensitive and heat stable (100 degrees C for 10 min). Escherichia coli heat-labile enterotoxin antibodies were measured in the colostrum samples by enzyme-linked immunosorbent assay. No E. coli enterotoxin-specific immunoglobulin A was detected, possibly due to the high background caused by the nonspecific adsorption of immunoglobulin A to the enzyme-linked immunosorbent assay plates in the absence of toxin. A total of 5 of 15 Ethiopian colostrum samples and 0 of 11 Norwegian colostrum samples neutralized the effect of E. coli heat-labile enterotoxin on YI adrenal cells. Both the Ethiopian and the Norwegian colostrum samples contained a non-immunoglobulin enterotoxin-inhibitory activity when the toxin was measured by enzyme-linked immunosorbent assay. This inhibitory activity was not trypsin sensitive, and extraction by chloroform-methanol indicated that the inhibitor was of a lipid nature. PMID:6268544

  8. Respective contributions to protection of primary and booster immunization with Escherichia coli heat-labile enterotoxin in rats.

    PubMed Central

    Klipstein, F A; Engert, R F

    1981-01-01

    The respective contributions to protection of the route and dosage of primary and booster immunizations with Escherichia coli heat-labile enterotoxin were evaluated in rats. The degree of protection was determined by challenge with toxin and viable bacteria in ligated ileal loops, and the serum antitoxin response was assayed by enzyme-linked immunosorbent assay. Primary immunization was effective only when given by the parenteral route. The degree of protection was enhanced a fivefold dosage increase in the primary parenteral immunization in rats given constant dosages of booster immunizations either parenterally or perorally, but not by further dosage increases. In contrast, the degree of protection rose when dosages of the booster immunizations were increased over a 25-fold range. Four weekly peroral, but only two biweekly parenteral, booster immunizations were necessary to achieve strong protection; biweekly combined parenteral and peroral booster immunizations yielded both strong, immediate and extended protection. The degree of protection against the toxin correlated with that against viable bacteria and with elevated serum antitoxin titers: all seven groups with a protection index of greater than 5 against the toxin had strong protection against heat-labile toxin-producing strains and fourfold or greater increases in the antitoxin titers, whereas none of the nine groups with a protection index of less than 3 was protected against bacteria or had an equivalent antitoxin response. These observations show that once an adequate parenteral primary immunization is given, the degree of protection is influenced primarily by the dosage of the booster immunizations, the necessary number of which is dependent on their route of administration. PMID:7011992

  9. Heat-labile Enterotoxins as Adjuvants or Anti-Inflammatory Agents

    PubMed Central

    Liang, Shuang; Hajishengallis, George

    2010-01-01

    Escherichia coli and Vibrio cholerae produce structurally related AB5-type heat-labile enterotoxins which are classified into two major types. The Type I subfamily includes cholera toxin and E. coli LT-I, whereas the Type II subfamily comprises LT-IIa and LT-IIb. In addition to their roles in microbial pathogenesis, the enterotoxins are widely and intensively studied for their exceptionally strong adjuvant and immunomodulatory activities, which are not necessarily dependent upon their abilities to elevate intracellular cAMP levels. Despite general structural similarities, these molecules, in intact or derivative form, display notable differences in their interactions with gangliosides or Toll-like receptors. This divergence results in differential immune response outcomes, the underlying mechanisms of which remain largely uncharacterized. Whereas the study of these molecules has been pivotal in understanding basic mechanisms of immune regulation, a formidable challenge is to dissociate toxicity from useful properties that can be exploited in vaccine development or for the treatment of autoimmune inflammatory diseases. PMID:20461887

  10. Rapid diagnosis of "Escherichia coli" heat labile enterotoxin (LT) by a coagglutination test.

    PubMed

    Wadstrm, T; Rnnberg, B

    1983-01-01

    Protein A positive Staphylococcus aureus cells, strain Cowan 1, were coated with a high titer anti-LT serum produced in rabbits. Purified E. coli heat labile enterotoxin (LT) was detected in a slide coagglutination (coa-LT) test with anti-LT coated staphylococci at the nanogram level (0.1-1 ng) while cholera toxin (CT) was detected only at much higher concentrations (10-50 ng). This LT immunoassay was evaluated in different modifications on enterotoxigenic E. coli (ETEC) isolated from humans and animal stool cultures. Colonies from primary stool cultures on blood agar were suspended and lysed in saline containing polymyxin B and a detergent (Triton X100), centrifuged and assayed for LT. The coa-LT and CHO-cell tests regularly detected LT among human LT producing ETEC, while ETEC strains from piglets and calves usually gave only weak reactions in the standard coa-LT test. We conclude that this coa-LT test is accurate and sensitive enough to be used in routine diagnosis of LT producing ETEC strains in stools from human patients. This test requires no special laboratory equipment and is well suited for rapid diagnosis of ETEC in small hospital laboratories and in developing countries. Heat stable enterotoxin (ST) giving a positive suckling mouse test, has been purified by hydrophobic interaction chromatography (HIC) on Octyl Sepharose. Purified ST was covalently linked to different proteins and such carrier complexes used for immunization of rabbits. The ST antibody titers obtained by various ST-carriers were compared in a ST coagglutination test (ST-coa) and in a conventional ELISA microtiter assay. The results and detection limits for ST in polymyxin-detergent cell lysates in human ETEC strains producing LT-ST or ST only will be presented. PMID:6347757

  11. Type II heat-labile enterotoxin of Escherichia coli activates adenylate cyclase in human fibroblasts by ADP ribosylation.

    PubMed

    Chang, P P; Moss, J; Twiddy, E M; Holmes, R K

    1987-08-01

    Type II heat-labile enterotoxin (LT-II) from Escherichia coli causes characteristic morphological changes and accumulation of cyclic AMP in Y-1 adrenal cells, but it is not neutralized by antisera against choleragen (CT) or the classical type I heat-labile enterotoxin (LT-1) from E. coli. The action of purified LT-II on CT- and LT-I-responsive human fibroblasts was investigated and compared with that of CT. Fibroblasts incubated with LT-II or CT had an increased cyclic AMP content as well as a fourfold elevation of membrane adenylate cyclase activity. In membranes, activation of cyclase by toxin was enhanced by NAD, GTP, and dithiothreitol. The effect of LT-II on intact fibroblasts or membranes was increased by trypsin treatment of toxin. Since activation of adenylate cyclase by LT-II was stimulated by NAD, the ability of LT-II to catalyze the [32P]ADP-ribosylation of membrane proteins in the presence of [32P]NAD from control and LT-II- and CT-treated fibroblasts was investigated. Similar proteins were [32P]ADP-ribosylated in membranes exposed to LT-II or CT; LT-II- and CT-specific labeling was significantly decreased in membranes prepared from cells preincubated with either LT-II or CT. These studies are consistent with the hypothesis that LT-II, similar to CT and LT-I, increases cyclic AMP by activating adenylate cyclase through the GTP-dependent ADP-ribosylation of specific membrane proteins. PMID:3112012

  12. Dissociation of Escherichia coli heat-labile enterotoxin adjuvanticity from ADP-ribosyltransferase activity.

    PubMed Central

    Dickinson, B L; Clements, J D

    1995-01-01

    The heat-labile enterotoxin (LT) of Escherichia coli is immunologically and physiochemically related to cholera enterotoxin. A number of studies have been performed to determine the relationship of the ADP-ribosylating enzymatic activity of these enterotoxins to toxicity and adjuvanticity. These studies have generally examined the effect of abolishing the ADP-ribosyltransferase activity of A1 by a variety of chemical or genetic manipulations. In every case, loss of enzymatic activity was associated with loss of biological activity and also with the ability of the molecules to function as oral adjuvants. Consequently, we explored an alternate approach to detoxification of LT without altering its adjuvanticity. Specifically, we generated a novel mutant form of LT by genetic modification of the proteolytically sensitive residues that join the A1 and A2 components of the A subunit. This mutant contains a single amino acid substitution within the disulfide subtended region joining A1 and A2. This mutant toxin, designated LT(R192G), is not sensitive to proteolytic activation, has negligible activity on mouse Y-1 adrenal tumor cells, and is devoid of ADP-ribosyltransferase activity. Nonetheless, LT(R192G) retains the ability to function as a mucosal adjuvant, increasing the serum immunoglobulin G (IgG) and mucosal IgA responses to coadministered antigen (OVA) beyond that achieved with administration of that antigen alone. Further, LT(R192G) prevented the induction of tolerance to coadministered antigen and did not induce tolerance against itself, as demonstrated by the presence of significant serum anti-LT IgG and mucosal anti-LT IgA antibodies in immunized mice. PMID:7729864

  13. Role of trypsin-like cleavage at arginine 192 in the enzymatic and cytotonic activities of Escherichia coli heat-labile enterotoxin.

    PubMed Central

    Grant, C C; Messer, R J; Cieplak, W

    1994-01-01

    Previous studies of cholera toxin and Escherichia coli heat-labile enterotoxin have suggested that proteolytic cleavage plays an important role in the expression of ADP-ribosyltransferase activity and toxicity. Specifically, several studies have implicated a trypsin-like cleavage at arginine 192, which lies within an exposed region subtended by a disulfide bond in the intact A subunit, in toxicity. To investigate the role of this modification in the enzymatic and cytotonic properties of heat-labile enterotoxin, the response of purified, recombinant A subunit to tryptic activation and the effect of substituting arginine 192 with glycine on the activities of the holotoxin were examined. The recombinant A subunit of heat-labile enterotoxin exhibited significant levels of ADP-ribosyltransferase activity that were only nominally increased (approximately twofold) by prior limited trypsinolysis. The enzymatic activity also did not appear to be affected by auto-ADP-ribosylation that occurs during the high-level synthesis of the recombinant A subunit in E. coli. A mutant form of the holotoxin containing the arginine 192-to-glycine substitution exhibited levels of cytotonic activity for CHO cells that were similar to that of the untreated, wild-type holotoxin but exhibited a marked delay in the ability to increase intracellular levels of cyclic AMP in Caco-2 cells. The results indicate that trypsin-like cleavage of the A subunit of E. coli heat-labile enterotoxin at arginine 192 is not requisite to the expression of enzymatic activity by the A subunit and further reveal that this modification, although it enhances the biological and enzymatic activities of the toxin, is not absolutely required for the enterotoxin to elicit cytotonic effects. Images PMID:7927684

  14. NAD-dependent ADP-ribosylation of arginine and proteins by Escherichia coli heat-labile enterotoxin.

    PubMed

    Moss, J; Garrison, S; Oppenheimer, N J; Richardson, S H

    1979-07-25

    Escherichia coli heat-labile enterotoxin (labile toxin, LT) catalyzed the hydrolysis of NAD to ADP-ribose and nicotinamide and the ADP-ribosylation of arginine (Moss, J., and Richardson, S.H. (1978) J. Clin. Invest. 62, 281-285). Analysis of the product of the ADP-ribosylation of arginine by nuclear magnetic resonance spectroscopy indicated that the reaction was stereospecific and resulted in the formation of alpha-ADP-ribosyl-L-arginine. This reaction product rapidly anomerized to yield a mixture of the alpha and beta forms. In the presence of [adenine-U-14C]NAD, E. coli enterotoxin catalyzed the transfer of the radiolabel to proteins; the ADP-ribosylation of proteins was inhibited by arginine methyl ester, an alternative substrate. Digestion of the 14C-protein with snake venom phosphodiesterase released predominantly 5'-AMP. No product was obtained with a mobility similar to that of 2'-(5''-phosphoribosyl)-5'-AMP. This result is consistent with the covalent attachment by the enterotoxin of ADP-ribose rather than poly(ADP-ribose) to protein. Thus, LT is catalytically equivalent to choleragen, an enterotoxin of Vibrio cholerae, and activates adenylate cyclase through a similar stereospecific ADP-ribosylation reaction. PMID:221495

  15. Simultaneous Exposure to Escherichia coli Heat-Labile and Heat-Stable Enterotoxins Increases Fluid Secretion and Alters Cyclic Nucleotide and Cytokine Production by Intestinal Epithelial Cells

    PubMed Central

    Read, Lisa T.; Hahn, Rachel W.; Thompson, Carli C.; Bauer, David L.; Norton, Elizabeth B.

    2014-01-01

    Enterotoxigenic Escherichia coli (ETEC) is a significant cause of diarrheal disease and death, especially in children in developing countries. ETEC causes disease by colonizing the small intestine and producing heat-labile toxin (LT), heat-stable toxin (ST), or both LT and ST (LT+ST). The majority of ETEC strains produce both ST and LT. Despite the prevalence of LT+ST-producing organisms, few studies have examined the physiologic or immunologic consequences of simultaneous exposure to these two potent enterotoxins. In the current report, we demonstrate that when LT and ST are both present, they increase water movement into the intestinal lumen over and above the levels observed with either toxin alone. As expected, cultured intestinal epithelial cells increased their expression of intracellular cyclic GMP (cGMP) when treated with ST and their expression of intracellular cyclic AMP (cAMP) when treated with LT. When both toxins were present, cGMP levels but not cAMP levels were synergistically elevated compared with the levels of expression caused by the corresponding single-toxin treatment. Our data also demonstrate that the levels of inflammatory cytokines produced by intestinal epithelial cells in response to LT are significantly reduced in animals exposed to both enterotoxins. These findings suggest that there may be complex differences between the epithelial cell intoxication and, potentially, secretory outcomes induced by ETEC strains expressing LT+ST compared with strains that express LT or ST only. Our results also reveal a novel mechanism wherein ST production may reduce the hosts' ability to mount an effective innate or adaptive immune response to infecting organisms. PMID:25287923

  16. Parvalbumin, labile heat and slowing of relaxation in mouse soleus and extensor digitorum longus muscles.

    PubMed Central

    Berquin, A; Lebacq, J

    1992-01-01

    1. Parvalbumin content, heat rate and rate of relaxation were measured in two mouse muscles: the slow-twitch soleus and the fast-twitch extensor digitorum longus (EDL). 2. No trace of parvalbumin was found in the soleus; EDL contained a mean of 4.86 mg of this protein per gram of fresh muscle (S.D. = 1.25). 3. Heat rate during 7 s isometric tetani in isolated soleus muscle at 20 degrees C can be described by the sum of an exponentially decaying term and a constant term. The exponential term is reduced by 67% in a second tetanus performed 1 s after a first one; its repriming is complete after a resting period of about 1 min. The exponential term has therefore the properties of labile heat. 4. Relaxation rate measured during 15 s of isometric interrupted tetani at 20 degrees C is nearly constant in the soleus, but decreases continuously with increasing tetanus duration in the EDL. In the latter, isometric tension also decreases continuously. 5. Therefore, parvalbumin can account neither for the labile heat production in mouse soleus nor for the slowing of relaxation associated with muscle fatigue observed after a few seconds of tetanus in EDL. The role of parvalbumin in striated muscles is thus reassessed, and other possible causes of labile heat production and slowing of relaxation are discussed. Images Fig. 1 PMID:1501147

  17. Genetic Rearrangements of the Regions Adjacent to Genes Encoding Heat-Labile Enterotoxins (eltAB) of Enterotoxigenic Escherichia coli Strains

    PubMed Central

    Schlör, Stefan; Riedl, Sabine; Blaß, Julia; Reidl, Joachim

    2000-01-01

    One of the most common bacterially mediated diarrheal infections is caused by enterotoxigenic Escherichia coli (ETEC) strains. ETEC-derived plasmids are responsible for the distribution of the genes encoding the main toxins, namely, the heat-labile and heat-stable enterotoxins. The origins and transfer modes (intra- or interplasmid) of the toxin-encoding genes have not been characterized in detail. In this study, we investigated the DNA regions located near the heat-labile enterotoxin-encoding genes (eltAB) of several clinical isolates. It was found that the eltAB region is flanked by conserved 236- and 280-bp regions, followed by highly variable DNA sequences which consist mainly of partial insertion sequence (IS) elements. Furthermore, we demonstrated that rearrangements of the eltAB region of one particular isolate, which harbors an IS91R sequence next to eltAB, could be produced by a recA-independent but IS91 sequence-dependent mechanism. Possible mechanisms of dissemination of IS element-associated enterotoxin-encoding genes are discussed. PMID:10618247

  18. Antibodies to heat-labile Escherichia coli enterotoxin in Apaches in Whiteriver, Arizona.

    PubMed Central

    Sack, R B; Hirschborn, N; Woodward, W E; Sack, D A; Cash, R A

    1975-01-01

    Antitoxin titers to heat-labile Escherichia coli enterotoxin were measured in Apache children hospitalized with acute diarrhea and in Apaches of different age groups without diarrhea in Whiteriver, Ariz. The study suggests that in this locale, exposure to enterotocigenic E. coli is probably widespread and occurs early in life. Antitoxin titer rises after idarrheal disease associated with enterotocigenic E. coli infection, however, were not regulary found. PMID:1107221

  19. Solid-Phase Microtiter Radioimmunoassay Blocking Test for Detection of Antibodies to Escherichia coli Heat-Labile Enterotoxin

    PubMed Central

    Greenberg, Harry B.; Levine, Myron M.; Merson, Michael H.; Sack, R. Bradley; Sack, David A.; Valdesuso, Jose R.; Nalin, David; Hoover, David; Chanock, Robert M.; Kapikian, Albert Z.

    1979-01-01

    The development of a solid-phase microtiter radioimmunoassay blocking test to detect serum antibody to Escherichia coli heat-labile enterotoxin is described. The assay is easy to perform and quantitate, and it is sensitive and specific. PMID:372216

  20. Solid-phase microtiter radioimmunoassay blocking test for detection of antibodies to Escherichia coli heat-labile enterotoxin.

    PubMed

    Greenberg, H B; Levine, M M; Merson, M H; Sack, R B; Sack, D A; Valdesuso, J R; Nalin, D; Hoover, D; Chanock, R M; Kapikian, A Z

    1979-01-01

    The development of a solid-phase microtiter radioimmunoassay blocking test to detect serum antibody to Escherichia coli heat-labile enterotoxin is described. The assay is easy to perform and quantitate, and it is sensitive and specific. PMID:372216

  1. Simulation of protein-sugar interactions: a computational model of the complex between ganglioside GM1 and the heat-labile enterotoxin of Escherichia coli.

    PubMed

    Bernardi, A; Raimondi, L; Zuccotto, F

    1997-06-01

    The cholera toxin from Vibrio cholerae (CT) and the 80% homologous heat-labile toxin of Escherichia coli (LT) are two well-known cases of sugar-binding proteins. The GM1:toxin complexes were chosen as test cases for the elaboration of a computational approach to the modeling of protein-saccharide interactions. The reliability of the method was evaluated on the LT:lactose complex. A model of this complex was built by performing a MC/EM conformational search of the sugar moiety within the binding pocket of LT, using the AMBER* force field and the GB/SA solvation model. The results are a reasonable reproduction of the reported X-ray structure of the complex. The same protocol was then applied to the LT:GM1 complex. The calculations were performed on a substructure that includes the protein shell within 5 A from GM1, three water molecules solvating Glu-51 carboxylate, and two water molecules at crystallographic sites 2 and 3. A satisfactory agreement was found with the recently published X-ray structure of the CT:GM1 complex. All the relevant interactions between the sugar and the residues involved in binding are well reproduced by the calculations. These results suggest that the substructure here identified can be taken as a realistic representation of the toxin binding surface and that the method presented in this paper can be used as a predictive tool in designing artificial LT (CT) binders and thus potential anticholera drugs. PMID:9191962

  2. The Escherichia coli heat-labile enterotoxin induces apoptosis of immature lymphocytes in vivo via a glucocorticoid-dependent pathway.

    PubMed

    Tamayo, Esther; Merino, Ramn; Gonzlez-Rojas, Jovanna; Marquina, Regina; Santiuste, Ins; Amado, Jos Antonio; Rappuoli, Rino; Del Giudice, Giuseppe; Merino, Jess

    2005-12-01

    Escherichia coli heat-labile enterotoxin (LT) exhibits a broad range of immunomodulatory activities, including the induction of lymphocyte-programmed cell death. However, the nature of the lymphoid populations sensitive to LT-induced apoptosis and the mechanisms used by this toxin to promote such activity are still unclear. In this study, we demonstrate that LT induces in mice a rapid increase in the levels of circulating corticosterone, resulting in a dramatic induction of cell death of immature CD4+CD8+, B220+IgM- and IgM+IgD- T and B cell progenitors, respectively. Apoptosis of these cell populations is similar to that reported after experimental treatment with corticosteroids, it is inhibited by mifepristone, a glucocorticoid receptor antagonist, and does not occur in adrenalectomized animals. These results clearly indicate that endogenous glucocorticoids are the mediators of the LT-induced cell death, which involves Bcl-2-dependent apoptotic pathways. The LT-mediated programmed cell death requires systemic exposure and the enzymatic activity of LT, since a mutant devoid of any enzymatic activity have no pro-apoptotic effect at any dose tested. PMID:16285009

  3. Type II Heat-Labile Enterotoxins from 50 Diverse Escherichia coli Isolates Belong Almost Exclusively to the LT-IIc Family and May Be Prophage Encoded

    PubMed Central

    Jobling, Michael G.; Holmes, Randall K.

    2012-01-01

    Some enterotoxigenic Escherichia coli (ETEC) produce a type II heat-labile enterotoxin (LT-II) that activates adenylate cyclase in susceptible cells but is not neutralized by antisera against cholera toxin or type I heat-labile enterotoxin (LT-I). LT-I variants encoded by plasmids in ETEC from humans and pigs have amino acid sequences that are ≥95% identical. In contrast, LT-II toxins are chromosomally encoded and are much more diverse. Early studies characterized LT-IIa and LT-IIb variants, but a novel LT-IIc was reported recently. Here we characterized the LT-II encoding loci from 48 additional ETEC isolates. Two encoded LT-IIa, none encoded LT-IIb, and 46 encoded highly related variants of LT-IIc. Phylogenetic analysis indicated that the predicted LT-IIc toxins encoded by these loci could be assigned to 6 subgroups. The loci corresponding to individual toxins within each subgroup had DNA sequences that were more than 99% identical. The LT-IIc subgroups appear to have arisen by multiple recombinational events between progenitor loci encoding LT-IIc1- and LT-IIc3-like variants. All loci from representative isolates encoding the LT-IIa, LT-IIb, and each subgroup of LT-IIc enterotoxins are preceded by highly-related genes that are between 80 and 93% identical to predicted phage lysozyme genes. DNA sequences immediately following the B genes differ considerably between toxin subgroups, but all are most closely related to genomic sequences found in predicted prophages. Together these data suggest that the LT-II loci are inserted into lambdoid type prophages that may or may not be infectious. These findings raise the possibility that production of LT-II enterotoxins by ETEC may be determined by phage conversion and may be activated by induction of prophage, in a manner similar to control of production of Shiga-like toxins by converting phages in isolates of enterohemmorhagic E. coli. PMID:22242186

  4. Evaluation of commercial antisera for serotyping heat-labile antigens of Campylobacter jejuni and Campylobacter coli.

    PubMed Central

    Nicholson, M A; Patton, C M

    1993-01-01

    Commercial antisera for serotyping 22 heat-labile antigens of Campylobacter jejuni and Campylobacter coli were evaluated by using 66 isolates from human and nonhuman sources. Test results were compared with results of tests using antisera produced at the Centers for Disease Control (CDC), Atlanta, Ga. All strains (three isolates of each of the 22 serotypes) were typeable with the CDC antisera. Of 66 test strains, 39 (59%) were typed as the same serotype with both sets of antisera. Twenty-four strains (36%), including two heat-labile serotype reference strains, were nonreactive with the commercial antisera, and three strains (4.5%) were typed as serotypes different from those obtained with CDC antisera. Five of the 22 commercial antisera correctly serotyped all homologous strains. Our study indicated that two polyvalent antiserum pools, 7 unabsorbed antisera, and 16 absorbed monovalent antisera are weak and need modification to enhance their antibody titers. Further studies are necessary to explain the antigenic change to a different serotype in three strains. PMID:8463402

  5. Hereditary heat-labile hexosaminidase B: its implication for recognizing Tay-Sachs genotypes.

    PubMed

    Navon, R; Nutman, J; Kopel, R; Gaber, L; Gadoth, N; Goldman, B; Nitzan, M

    1981-11-01

    Two pairs of alleles, at the two loci of hexosaminidase (HEX), were found to segregate in an Arab inbred family: the normal and the mutant Tay-Sachs (TSD) alleles of HEX A, and the normal and a mutant allele of HEX B. Since the mutant HEX B is heat labile, no reliable identification of TSD genotypes can be obtained in its presence, as long as the proportions of HEX A and B are estimated by the routinely used heat-inactivation method. The genotypes may be correctly identified in such cases by separation of the two isoenzymes on ion-exchange chromatography, estimating their individual activities, and calculating the ratio between them. Of the nine genotype combinations possible with these two pairs of alleles, five have been identified in the reported family by this procedure. PMID:6459736

  6. Context-Dependent Activation Kinetics Elicited by Soluble versus Outer Membrane Vesicle-Associated Heat-Labile Enterotoxin ▿

    PubMed Central

    Chutkan, Halima; Kuehn, Meta J.

    2011-01-01

    Enterotoxigenic Escherichia coli (ETEC) is the leading cause of traveler's diarrhea and children's diarrhea worldwide. Among its virulence factors, ETEC produces heat-labile enterotoxin (LT). Most secreted LT is associated with outer membrane vesicles that are rich in lipopolysaccharide. The majority of prior studies have focused on soluble LT purified from ETEC periplasm. We investigated the hypothesis that the extracellular vesicle context of toxin presentation might be important in eliciting immune responses. We compared the polarized epithelial cell responses to apically applied soluble LT and LT-containing vesicles (LT+ vesicles) as well as controls using a catalytically inactive mutant of LT and vesicles lacking LT. Although vesicle treatments with no or catalytically inactive LT induced a modest amount of interleukin-6 (IL-6), samples containing catalytically active LT elicited higher levels. A combination of soluble LT and LT-deficient vesicles induced significantly higher IL-6 levels than either LT or LT+ vesicles alone. The responses to LT+ vesicles were found to be independent of the canonical LT pathway, because the inhibition of cyclic AMP response element (CRE)-binding protein (CREB) phosphorylation did not lead to a decrease in cytokine gene expression levels. Furthermore, soluble LT caused earlier phosphorylation of CREB and activation of CRE compared with LT+ vesicles. Soluble LT also led to the activation of activator protein 1, whereas LT+ vesicle IL-6 responses appeared to be mediated by NF-κB. In summary, the results demonstrate that soluble LT and vesicle-bound LT elicit ultimately similar cytokine responses through distinct different activation pathways. PMID:21708992

  7. Escherichia coli heat-labile enterotoxin genes are flanked by repeated deoxyribonucleic acid sequences.

    PubMed Central

    Yamamoto, T; Yokota, T

    1981-01-01

    The enterotoxin regions of the heat-labile and heat-stable enterotoxin (LT+ ST+) plasmid, pJY11, originating in a clinically isolated Escherichia coli strain, have been isolated as various-sized deoxyribonucleic acid (DNA) fragments by using cloning vehicles. The structure of the LT+ region and its neighboring DNA regions was studied by utilizing these recombinant plasmids. The LT+ region consisted of at least two genes, toxA and toxB, which could complement each other in trans. The toxA- and toxB-encoded polypeptides (LT subunits A and B, respectively) were identified by their immunological cross-reactivity with Vibrio cholerae enterotoxin subunit A or B. These tox genes and the promoter(s) were localized with respect to the restriction endonuclease cleavage map. The LT+ region was flanked by repeated DNA sequences (designated as beta). Another tox gen(s), encoding ST (designated as toxS), which was also flanked by inverted, repeated DNA sequences (designated as alpha), was located between one of the beta sequences and the LT+ region. These novel DNA structures (beta-alpha-toxS-alpha-toxA-toxB-beta) suggest the possibility that the LT+ region is on a transposon containing an ST transposon within the structure. Images PMID:6257652

  8. An outbreak of gastroenteritis due to a heat-labile enterotoxin-producing strain of Escherichia coli.

    PubMed Central

    Wood, L V; Wolfe, W H; Ruiz-Palacios, G; Foshee, W S; Corman, L I; McCleskey, F; Wright, J A; DuPont, H L

    1983-01-01

    In April 1981, an outbreak of gastroenteritis, characterized by diarrhea and abdominal cramps, occurred in 282 of approximately 3,000 personnel at a large metropolitan hospital in San Antonio, Tex. There was a significant association between illness and eating at the hospital cafeteria (P = 0.0008), but no specific food could be incriminated. Stools or rectal swabs from 54 ill individuals produced almost pure cultures of Escherichia coli. Cultures from 51 of these subjects had identical antibiotic sensitivity patterns, and 38 had the same biotype. Isolates from 45 persons were tested for production of heat-stable and heat-labile enterotoxins, using the suckling mouse and Y-1 adrenal cell assays, respectively. Of 45 isolates, 41 produced heat-labile enterotoxins, while 0 of 45 produced heat-stabile enterotoxins. Two isolates were rough, and 34 of the remaining 43 were serotype O25:H-. Two strains were O25:H+. None of the 45 strains possessed hemagglutination patterns typical of colonization factor antigens I or II. Six of seven O25:H- heat-labile enterotoxin-positive strains selected at random were piliated as seen by electron microscopy but did not agglutinate with anti-colonization factor antigens I or II antisera. PMID:6350188

  9. GITR contributes to the systemic adjuvanticity of the Escherichia coli heat-labile enterotoxin.

    PubMed

    Tamayo, Esther; Postigo, Jorge; Gonzlez, Jovanna; Fernndez-Rey, Maigualida; Iglesias, Marcos; Santiuste, Ins; Riccardi, Carlo; Rappuoli, Rino; Del Giudice, Giuseppe; Merino, Ramn; Merino, Jess

    2010-03-01

    The Escherichia coli heat-labile enterotoxin (LT) possesses a powerful mucosal and systemic adjuvant effect. However, little is known about the cellular and molecular basis of the immunostimulatory activity of LT at the mucosal level, and even less information is available on the mechanisms underlying its systemic adjuvant activity. In this study, we show that distinct mechanisms are responsible for the parenteral and mucosal adjuvanticity of LT. Indeed, the systemic administration of LT upregulates the expression of glucocorticoid-induced TNFR-related protein (GITR), but not other activation markers, in naive T cells. Using WT and GITR-deficient mice and LT and its enzymatically inactive mutant LTK63 as adjuvants, we show that the induction of GITR expression in T cells accounts for the systemic immunostimulatory capacity of LT, which requires an intact enzymatic activity. In contrast, the mucosal administration of LT does not induce GITR expression on Peyer's patche T cells and accordingly no differences are observed in the mucosal adjuvanticity of LT between WT and GITR-deficient mice. Altogether, our results demonstrate the distinct effect of LT after parenteral administration when compared with the mucosal delivery, and describe a new mechanism of LT adjuvanticity related to its ability to induce the expression of GITR in CD4(+) T cells. PMID:20017194

  10. Thermo-osmoregulation of heat-labile enterotoxin expression by Escherichia coli.

    PubMed

    Trachman, Julie D; Yasmin, Munira

    2004-11-01

    Enterotoxigenic Escherichia coli causes diarrhea by producing several virulence factors including heat-labile enterotoxin (LT). LT is maximally expressed at 37 degrees C. The histone-like nucleoid structuring protein (H-NS) appears to inhibit LT expression by binding to a downstream regulatory element (DRE) at low temperatures. An hns+ E. coli strain, X7026, carrying an LT-beta-galactosidase translational fusion plasmid (pLT-lac) was shown to be responsive to varying amounts of sodium chloride (NaCl) as well as sucrose or lithium chloride. Maximal responsiveness to the various osmolytes was obtained with cells grown at 37 degrees C under microaerophilic conditions. Temperature-osmotic upshift experiments demonstrate LT expression is thermo-osmoregulated. pLT-lac was tested in an hns strain or its congenic hns+ strain for its response to NaCl. LT expression is elevated in the hns strain regardless of NaCl concentration and retains its osmoresponsiveness. The response of the DRE deletion plasmid (pLT-lacDeltaNC) to NaCl is similar to that of the undeleted plasmid. PMID:15486710

  11. Heat-labile enterotoxin of Escherichia coli promotes intestinal colonization of Salmonella enterica.

    PubMed

    Verbrugghe, Elin; Van Parys, Alexander; Leyman, Bregje; Boyen, Filip; Arnouts, Sven; Lundberg, Urban; Ducatelle, Richard; Van den Broeck, Wim; Yekta, Maryam Atef; Cox, Eric; Haesebrouck, Freddy; Pasmans, Frank

    2015-12-01

    Enterotoxigenic Escherichia coli (ETEC) is an important cause of infantile and travellers' diarrhoea, which poses a serious health burden, especially in developing countries. In addition, ETEC bacteria are a major cause of illness and death in neonatal and recently weaned pigs. The production of a heat-labile enterotoxin (LT) promotes the colonization and pathogenicity of ETEC and may exacerbate co-infections with other enteric pathogens such as Salmonella enterica. We showed that the intraintestinal presence of LT dramatically increased the intestinal Salmonella Typhimurium load in experimentally inoculated pigs. This could not be explained by direct alteration of the invasion or survival capacity of Salmonella in enterocytes, in vitro. However, we demonstrated that LT affects the enteric mucus layer composition in a mucus-secreting goblet cell line by significantly decreasing the expression of mucin 4. The current results show that LT alters the intestinal mucus composition and aggravates a Salmonella Typhimurium infection, which may result in the exacerbation of the diarrhoeal illness. PMID:26616654

  12. Polymyxin B-Induced Release of Low-Molecular-Weight, Heat-Labile Enterotoxin from Escherichia coli

    PubMed Central

    Evans, Doyle J.; Evans, Dolores G.; Gorbach, Sherwood L.

    1974-01-01

    Polymyxin B-induced release of enterotoxin from Escherichia coli strain H-10407 was demonstrated. Incubation of E. coli cells derived from 6-h cultures with polymyxin caused the rapid release of enterotoxin with a molecular weight of approximately 20,000, as estimated by the gel filtration technique. The rapidity of the release of enterotoxin indicates that it probably resides in the periplasmic space of the cell. The low-molecular-weight enterotoxin possessed vascular permeability factor and diarrheagenic activities, both of which were found to be heat-labile. The permeability factor activity of this enterotoxin was neutralized by antisera prepared against crude E. coli enterotoxin, Vibrio cholerae enterotoxin (choleragen), and V. cholerae toxoid (choleragenoid), respectively. Supernatant fluids of 6-h E. coli cultures did not contain this molecular form of enterotoxin but did contain very high-molecular-weight, heat-labile enterotoxin. Incubation of cells derived from older (18 h) cultures with polymyxin caused the release of both low- (20,000) and high-molecular-weight forms of enterotoxin. We concluded that either the 20,000-dalton form of heat-labile enterotoxin is not released by E. coli under in vitro growth conditions or that enterotoxin released in this form is rapidly destroyed or inactivated. PMID:16558081

  13. Activation of adenylate cyclase by heat-labile Escherichia coli enterotoxin. Evidence for ADP-ribosyltransferase activity similar to that of choleragen.

    PubMed

    Moss, J; Richardson, S H

    1978-08-01

    Highly purified, polymyxin-released, low molecular weight Escherichia coli heat-labile enterotoxin (LT) catalyzed the hydrolysis of NAD to ADP-ribose and nicotinamide. This NAD glycohydrolase activity was stimulated by dithiothreitol and was independent of cellular components. Nicotinamide formation was enhanced by arginine methyl ester > d-arginine congruent with l-arginine congruent with guanidine. A 20-fold increase in activity was noted with arginine methyl ester, and maximal activity again required dithiothreitol. When the reaction was initiated with toxin, a delay was observed before a constant rate was established. The reaction products found after incubation of [adenine-U-(14)C]NAD and l-[(3)H]arginine or unlabeled arginine methyl ester with the enterotoxin had mobilities on thin-layer chromatograms similar to the reaction products obtained after incubation of choleragen with these substrates and are consistent with the formation of ADP-ribose-l-arginine and ADP-ribose-l-arginine methyl ester, respectively. Both toxins, which catalyze the NAD-dependent activation of adenylate cyclase, thus appear to possess NAD glycohydrolase and ADP-ribosyltransferase activities. Although the activities of both toxins are dependent on dithiothreitol, Escherichia coli enterotoxin exhibited optimal activity in Tris (Cl(-)) (pH 7.5) and was inhibited by high concentrations of potassium phosphate (pH 7.0) or low pH (sodium acetate, pH 6.2). It appears that the optimal assay conditions as well as the kinetic constants for the reactants differ from those previously noted with choleragen. It is probable therefore that although the two toxins catalyze similar reactions, they differ in primary structure. The presence of transferase and glycohydrolase activities in structurally distinct toxins that activate adenylate cyclase strengthens our hypothesis that the ADP-ribosylation of arginine is a model for the NAD-dependent activation of adenylate cyclase; activation may result from ADP-ribosylation of the cyclase itself or of a protein that regulates its activity. PMID:209060

  14. Cross-protection of infant mice against intestinal colonisation by Campylobacter jejuni: importance of heat-labile serotyping (Lior) antigens.

    PubMed

    Abimiku, A G; Dolby, J M

    1988-08-01

    An association of the heat-labile antigens detected by the Lior serotyping scheme with ability to protect infant mice against gastrointestinal colonisation with Campylobacter jejuni has been established. Overall, 39 (57%) of 68 infant mice challenged with a heterologous strain of the same Lior serotype as the vaccine strain were protected, compared with 40 (85%) of 47 infants protected against a homologous challenge. In contrast, none of the infant mice challenged with a strain carrying the same heat-stable antigens (i.e., of the same Penner serotype as the vaccine strain) were protected. PMID:3398033

  15. Protein kinase activity from vaccinia virions: solubilization and separation into heat-labile and heat-stable components.

    PubMed

    Kleiman, J; Moss, B

    1973-10-01

    A protein kinase was solubilized from whole vaccinia virions by using a solution containing deoxycholate, dithiothreitol, and sodium or potassium chloride. The released enzyme was completely dependent on Mg(2+) and was greatly stimulated by added basic proteins such as protamine or histones. Dithiothreitol was also stimulatory, whereas GTP, CTP, UTP, and P(i) at concentrations equimolar with ATP had little or no effect. Attempts to purify the protein kinase were initially unsuccessful, leading us to consider that either the enzyme was extremely labile or that two readily separable components were required for activity. The observation that the material extracted with NP-40 detergent during the preparation of viral cores stimulated the protein kinase activity of the intact cores supported the second possibility. As the protein kinase, now solubilized from viral cores, was passed through successive DEAE-cellulose columns, it became increasingly dependent for activity on addition of the NP-40 extract. A 30- to 40-fold stimulation of protein kinase activity, which afforded recovery of essentially all starting activity, could be effected by addition of the NP-40 extract to the partially purified enzyme. The NP-40 extract was shown to contain a heat stable, trypsin-sensitive protein, whose action could not be duplicated by cyclic nucleotides. PMID:4776967

  16. Towards Rational Design of a Toxoid Vaccine against the Heat-Stable Toxin of Escherichia coli.

    PubMed

    Taxt, Arne M; Diaz, Yuleima; Aasland, Rein; Clements, John D; Nataro, James P; Sommerfelt, Halvor; Puntervoll, Pål

    2016-04-01

    EnterotoxigenicEscherichia coli(ETEC) is an important cause of diarrheal disease and death in children <5 years old. ETEC strains that express the heat-stable toxin (ST), with or without the heat-labile toxin, are among the four most important diarrhea-causing pathogens. This makes ST an attractive target for an ETEC vaccine. An ST vaccine should be nontoxic and elicit an immune response that neutralizes native ST without cross-reacting with the human endogenous guanylate cyclase C receptor ligands. To identify variants of ST with no or low toxicity, we screened a library of all 361 possible single-amino-acid mutant forms of ST by using the T84 cell assay. Moreover, we identified mutant variants with intact epitopes by screening for the ability to bind neutralizing anti-ST antibodies. ST mutant forms with no or low toxicity and intact epitopes are termed toxoid candidates, and the top 30 candidates all had mutations of residues A14, N12, and L9. The identification of nontoxic variants of L9 strongly suggests that it is a novel receptor-interacting residue, in addition to the previously identified N12, P13, and A14 residues. The screens also allowed us to map the epitopes of three neutralizing monoclonal antibodies, one of which cross-reacts with the human ligand uroguanylin. The common dominant epitope residue for all non-cross-reacting antibodies was Y19. Our results suggest that it should be possible to rationally design ST toxoids that elicit neutralizing immune responses against ST with minimal risk of immunological cross-reactivity. PMID:26883587

  17. Comparative Analysis of the Mucosal Adjuvanticity of the Type II Heat-Labile Enterotoxins LT-IIa and LT-IIb

    PubMed Central

    Martin, Michael; Metzger, Daniel J.; Michalek, Suzanne M.; Connell, Terry D.; Russell, Michael W.

    2000-01-01

    Cholera toxin (CT) and the heat-labile enterotoxin of Escherichia coli (LT-I) are members of the serogroup I heat-labile enterotoxins (HLT) and can serve as systemic and mucosal adjuvants. However, information is lacking with respect to the structurally related but antigenically distinct serogroup II HLT, LT-IIa and LT-IIb, which have different binding specificities for ganglioside receptors. The purpose of this study was to assess the effectiveness of LT-IIa and LT-IIb as mucosal adjuvants in comparison to the prototypical type I HLT, CT. BALB/c mice were immunized by the intranasal (i.n.) route with the surface protein adhesin AgI/II of Streptococcus mutans alone or supplemented with an adjuvant amount of CT, LT-IIa, or LT-IIb. Antigen-specific antibody responses in saliva, vaginal wash, and plasma were assayed by enzyme-linked immunosorbent assay. Mice given AgI/II with LT-IIa or LT-IIb by the i.n. route had significantly higher mucosal and systemic antibody responses than mice immunized with AgI/II alone. Anti-AgI/II immunoglobulin A (IgA) antibody activity in saliva and vaginal secretions of mice given AgI/II with LT-IIa or LT-IIb was statistically similar in magnitude to that seen in mice given AgI/II and CT. LT-IIb significantly enhanced the number of AgI/II-specific antibody-secreting cells in the draining superficial cervical lymph nodes compared to LT-IIa and CT. LT-IIb and CT induced significantly higher plasma anti-AgI/II IgG titers compared to LT-IIa. When LT-IIb was used as adjuvant, the proportion of plasma IgG2a relative to IgG1 anti-AgI/II antibody was elevated in contrast to the predominance of IgG1 antibodies promoted by AgI/II alone or when CT or LT-IIa was used. In vitro stimulation of AgI/II-specific cells from the superficial lymph nodes and spleen revealed that LT-IIa and LT-IIb induced secretion of interleukin-4 and significantly higher levels of gamma interferon compared to CT. These results demonstrate that the type II HLT LT-IIa and LT-IIb exhibit potent and distinct adjuvant properties for stimulating immune responses to a noncoupled protein immunogen after mucosal immunization. PMID:10603399

  18. Activation of Escherichia coli heat-labile enterotoxins by native and recombinant adenosine diphosphate-ribosylation factors, 20-kD guanine nucleotide-binding proteins.

    PubMed

    Lee, C M; Chang, P P; Tsai, S C; Adamik, R; Price, S R; Kunz, B C; Moss, J; Twiddy, E M; Holmes, R K

    1991-05-01

    Escherichia coli heat-labile enterotoxins (LT) are responsible in part for "traveler's diarrhea" and related diarrheal illnesses. The family of LTs comprises two serogroups termed LT-I and LT-II; each serogroup includes two or more antigenic variants. The effects of LTs result from ADP ribosylation of Gs alpha, a stimulatory component of adenylyl cyclase; the mechanism of action is identical to that of cholera toxin (CT). The ADP-ribosyltransferase activity of CT is enhanced by 20-kD guanine nucleotide-binding proteins, known as ADP-ribosylation factors or ARFs. These proteins directly activate the CTA1 catalytic unit and stimulate its ADP ribosylation of Gs alpha, other proteins, and simple guanidino compounds (e.g., agmatine). Because of the similarities between CT and LTs, we investigated the effects of purified bovine brain ARF and a recombinant form of bovine ARF synthesized in Escherichia coli on LT activity. ARF enhanced the LT-I-, LT-IIa-, and LT-IIb-catalyzed ADP ribosylation of agmatine, as well as the auto-ADP ribosylation of the toxin catalytic unit. Stimulation of ADP-ribosylagmatine formation by LTs and CT in the presence of ARF was GTP dependent and enhanced by sodium dodecyl sulfate. With agmatine as substrate, LT-IIa and LT-IIb exhibited less than 1% the activity of CT and LT-Ih. CT and LTs catalyzed ADP-ribosyl-Gs alpha formation in a reaction dependent on ARF, GTP, and dimyristoyl phosphatidylcholine/cholate. With Gs alpha as substrate, the ADP-ribosyltransferase activities of the toxins were similar, although CT and LT-Ih appeared to be slightly more active than LT-IIa and LT-IIb. Thus, LT-IIa and LT-IIb appear to differ somewhat from CT and LT-Ih in substrate specificity. Responsiveness to stimulation by ARF, GTP, and phospholipid/detergent as well as the specificity of ADP-ribosyltransferase activity are functions of LTs from serogroups LT-I and LT-II that are shared with CT. PMID:1902492

  19. Ginger and its bioactive component inhibit enterotoxigenic Escherichia coli heat-labile enterotoxin-induced diarrhea in mice.

    PubMed

    Chen, Jaw-Chyun; Huang, Li-Jiau; Wu, Shih-Lu; Kuo, Sheng-Chu; Ho, Tin-Yun; Hsiang, Chien-Yun

    2007-10-17

    Ginger is one of the most commonly used fresh herbs and spices. Enterotoxigenic Escherichia coli heat-labile enterotoxin (LT)-induced diarrhea is the leading cause of infant death in developing countries. In this study, we demonstrated that ginger significantly blocked the binding of LT to cell-surface receptor G M1, resulting in the inhibition of fluid accumulation in the closed ileal loops of mice. Biological-activity-guided searching for active components showed that zingerone (vanillylacetone) was the likely active constituent responsible for the antidiarrheal efficacy of ginger. Further analysis of chemically synthesized zingerone derivatives revealed that compound 31 (2-[(4-methoxybenzyl)oxy]benzoic acid) significantly suppressed LT-induced diarrhea in mice via an excellent surface complementarity with the B subunits of LT. In conclusion, our findings provide evidence that ginger and its derivatives may be effective herbal supplements for the clinical treatment of enterotoxigenic Escherichia coli diarrhea. PMID:17880155

  20. Repair of radiation-induced heat-labile sites is independent of DNA-PKcs, XRCC1 or PARP

    SciTech Connect

    Stenerlöw, Bo; Karlsson, Karin H.; Radulescu, Irina; Rydberg, Bjorn; Stenerlow, Bo

    2008-04-29

    Ionizing radiation induces a variety of different DNA lesions: in addition to the most critical DNA damage, the DSB, numerous base alterations, SSBs and other modifications of the DNA double-helix are formed. When several non-DSB lesions are clustered within a short distance along DNA, or close to a DSB, they may interfere with the repair of DSBs and affect the measurement of DSB induction and repair. We have previously shown that a substantial fraction of DSBs measured by pulsed-field gel electrophoresis (PFGE) are in fact due to heat-labile sites (HLS) within clustered lesions, thus reflecting an artifact of preparation of genomic DNA at elevated temperature. To further characterize the influence of HLS on DSB induction and repair, four human cell lines (GM5758, GM7166, M059K, U-1810) with apparently normal DSB rejoining were tested for bi-phasic rejoining after gamma irradiation. When heat-released DSBs were excluded from the measurements the fraction of fast rejoining decreased to less than 50% of the total. However, neither the half-times of the fast (t{sub 1/2} = 7-8 min) or slow (t{sub 1/2} = 2.5 h) DSB rejoining were changed significantly. At t=0 the heat-released DSBs accounted for almost 40% of the DSBs, corresponding to 10 extra DSB/cell/Gy in the initial DSB yield. These heat-released DSBs were repaired within 60-90 min in all tested cells, including M059K cells treated with wortmannin or DNA-PKcs defect M059J cells. Furthermore, cells lacking XRCC1 or Poly(ADP-ribose) polymerase-1 (PARP-1) rejoined both total DSBs and heat-released DSBs similar to normal cells. In summary, the presence of heat-labile sites have a substantial impact on DSB induction yields and DSB rejoining rates measured by pulsed-field gel electrophoresis, and HLS repair is independent of DNA-PKcs, XRCC1 and PARP.

  1. Intermolecular interactions between the A and B subunits of heat-labile enterotoxin from Escherichia coli promote holotoxin assembly and stability in vivo.

    PubMed Central

    Streatfield, S J; Sandkvist, M; Sixma, T K; Bagdasarian, M; Hol, W G; Hirst, T R

    1992-01-01

    Cholera toxin and the related heat-labile enterotoxin (LT) produced by Escherichia coli consist of a holotoxin of one A subunit and five B subunits (AB5). Here we investigate the domains of the A subunit (EtxA) of E. coli LT which influence the events of B-subunit (EtxB) oligomerization and the formation of a stable AB5 holotoxin complex. We show that the C-terminal 14 amino acids of the A subunit comprise two functional domains that differentially affect oligomerization and holotoxin stability. Deletion of the last 14 amino acids (-14) from the A subunit resulted in a molecule that was significantly impaired in its capacity to promote the assembly of a mutant B subunit, EtxB191.5. In contrast, deletion of the last four amino acids (-4) from the A subunit gave a molecule that retained such a capacity. This suggests that C-terminal residues within the -14 to -4 region of the A subunit are important for promoting the oligomerization of EtxB. In addition, we demonstrate that the truncated A subunit lacking the last 4 amino acids was unable to form a stable AB5 holotoxin complex even though it promoted B-subunit oligomerization. This suggests that the last 4 residues of the A subunit function as an "anchoring" sequence responsible for maintaining the stability of A/B subunit interaction during holotoxin assembly. These data represent an important example of how intermolecular interactions between polypeptides in vivo can modulate the folding and assembly of a macromolecular complex. Images PMID:1465452

  2. Effects of Site-Directed Mutagenesis of Escherichia coli Heat-Labile Enterotoxin on ADP-Ribosyltransferase Activity and Interaction with ADP-Ribosylation Factors

    PubMed Central

    A. Stevens, Linda; Moss, Joel; Vaughan, Martha; Pizza, Mariagrazia; Rappuoli, Rino

    1999-01-01

    Escherichia coli heat-labile enterotoxin (LT), an oligomeric protein with one A subunit (LTA) and five B subunits, exerts its effects via the ADP-ribosylation of Gsα, a guanine nucleotide-binding (G) protein that activates adenylyl cyclase. LTA also ADP-ribosylates simple guanidino compounds (e.g., arginine) and catalyzes its own auto-ADP-ribosylation. All LTA-catalyzed reactions are enhanced by ADP-ribosylation factors (ARFs), 20-kDa guanine nucleotide-binding proteins. Replacement of arginine-7 (R7K), valine-53 (V53D), serine-63 (S63K), valine 97 (V97K), or tyrosine-104 (Y104K) in LTA resulted in fully assembled but nontoxic proteins. S63K, V53D, and R7K are catalytic-site mutations, whereas V97K and Y104K are amino acid replacements adjacent to and outside of the catalytic site, respectively. The effects of mutagenesis were quantified by measuring ADP-ribosyltransferase activity (i.e., auto-ADP-ribosylation and ADP-ribosylagmatine synthesis) and interaction with ARF (i.e., inhibition of ARF-stimulated cholera toxin ADP-ribosyltransferase activity and effects of ARF on mutant auto-ADP-ribosylation). All mutants were inactive in the ADP-ribosyltransferase assay; however, auto-ADP-ribosylation in the presence of recombinant human ARF6 was detected, albeit much less than that of native LT (Y104K > V53D > V97K > R7K, S63K). Based on the lack of inhibition by free ADP-ribose, the observed auto-ADP-ribosylation activity was enzymatic and not due to the nonenzymatic addition of free ADP-ribose. V53D, S63K, and R7K were more effective than Y104K or V97K in blocking ARF stimulation of cholera toxin ADP-ribosyltransferase. Based on these data, it appears that ARF-binding and catalytic sites are not identical and that a region outside the NAD cleft may participate in the LTA-ARF interaction. PMID:9864224

  3. Effects of site-directed mutagenesis of Escherichia coli heat-labile enterotoxin on ADP-ribosyltransferase activity and interaction with ADP-ribosylation factors.

    PubMed

    Stevens, L A; Moss, J; Vaughan, M; Pizza, M; Rappuoli, R

    1999-01-01

    Escherichia coli heat-labile enterotoxin (LT), an oligomeric protein with one A subunit (LTA) and five B subunits, exerts its effects via the ADP-ribosylation of Gsalpha, a guanine nucleotide-binding (G) protein that activates adenylyl cyclase. LTA also ADP-ribosylates simple guanidino compounds (e.g., arginine) and catalyzes its own auto-ADP-ribosylation. All LTA-catalyzed reactions are enhanced by ADP-ribosylation factors (ARFs), 20-kDa guanine nucleotide-binding proteins. Replacement of arginine-7 (R7K), valine-53 (V53D), serine-63 (S63K), valine 97 (V97K), or tyrosine-104 (Y104K) in LTA resulted in fully assembled but nontoxic proteins. S63K, V53D, and R7K are catalytic-site mutations, whereas V97K and Y104K are amino acid replacements adjacent to and outside of the catalytic site, respectively. The effects of mutagenesis were quantified by measuring ADP-ribosyltransferase activity (i.e., auto-ADP-ribosylation and ADP-ribosylagmatine synthesis) and interaction with ARF (i.e., inhibition of ARF-stimulated cholera toxin ADP-ribosyltransferase activity and effects of ARF on mutant auto-ADP-ribosylation). All mutants were inactive in the ADP-ribosyltransferase assay; however, auto-ADP-ribosylation in the presence of recombinant human ARF6 was detected, albeit much less than that of native LT (Y104K > V53D > V97K > R7K, S63K). Based on the lack of inhibition by free ADP-ribose, the observed auto-ADP-ribosylation activity was enzymatic and not due to the nonenzymatic addition of free ADP-ribose. V53D, S63K, and R7K were more effective than Y104K or V97K in blocking ARF stimulation of cholera toxin ADP-ribosyltransferase. Based on these data, it appears that ARF-binding and catalytic sites are not identical and that a region outside the NAD cleft may participate in the LTA-ARF interaction. PMID:9864224

  4. Evaluation of heat-labile enterotoxins type IIa and type IIb in the pathogenicity of enterotoxigenic Escherichia coli for neonatal pigs

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Type II heat-labile enterotoxins (LT-II) have been reported in Escherichia coli isolates from humans, animals, food and water samples. The roles of the antigenically distinguishable LT-IIa and LT-IIb subtypes in pathogenesis and virulence of enterotoxigenic E. coli (ETEC) have not been previously re...

  5. The B subunit of Escherichia coli heat labile enterotoxin abrogates oral tolerance, promoting predominantly Th2-type immune responses.

    PubMed

    Plant, Andrea; Williams, Rachel; Jackson, Michelle E; Williams, Neil A

    2003-11-01

    Mucosal antigen encounter usually results in a state of systemic non-responsiveness (tolerance). This failure to mount a protective response is a major hurdle to mucosal vaccine development. Hence, the identification of safe and effective mucosal adjuvants promoting protective immunity is of critical importance. The non-toxic B subunit of Escherichia coli heat labile enterotoxin(EtxB) is a potent nasal adjuvant; however, its usefulness following oral delivery is unconfirmed. We used DO11.10 chimeric mice to assess whether EtxB could abrogate tolerance to oral OVA. We show that admixing EtxB with OVA for oral immunization abrogates oral tolerance and results in a weak anti-OVA immune response. Importantly, EtxB profoundly modulated the nature of the response to subsequent parenteral challenge, promoting IgG1 in favor of IgG2a antibodies and depressing IFN-gamma production while elevating TGF-beta secretion. The addition of EtxB promoted T cell division, as assessed by loss of staining with carboxyfluorescein diacetate succinimidyl ester. Enhanced cell division promoted by EtxB was associated with T cell differentiation (increased numbers of CD45RBlow cells) in vivo, although dividing OVA-specific T cells were CD25-. These data suggest that although EtxB is a weak oral adjuvant, it can profoundly modulate the nature of the immune response to admixed antigen. PMID:14579287

  6. Deletion mutations in N-terminal α1 helix render heat labile enterotoxin B subunit susceptible to degradation

    PubMed Central

    Alone, Pankaj V.; Malik, Gunjan; Krishnan, Anuja; Garg, Lalit C.

    2007-01-01

    Heat-labile enterotoxin (LT) from enterotoxigenic Escherichia coli is a heterohexameric protein consisting of an enzymatically active A subunit, LTA, and a carrier pentameric B subunit, LTB. It is clear from the crystal structure of LTB that the N-terminal α1 helix lies outside the core structure. However, the function of the N-terminal α1 helix of LTB is unknown. The present work was carried out to investigate the effect of site-directed mutagenesis of the α1 helix on LTB synthesis. Six amino acids (PQSITE) located at positions 2–7 from the N terminus, including 4 aa from the α1 helix, were deleted by site-directed mutagenesis. The deletion resulted in complete inhibition of LTB expression in E. coli when expressed along with its signal sequence. A single amino acid deletion within the α1 helix also resulted in loss of expression. However, a single amino acid deletion outside the α1 helix did not affect LTB synthesis. Mutant proteins, whose synthesis was not detected in vivo, could be successfully translated in vitro by using the coupled transcription–translation system. Immunoblot analysis, Northern blot analysis, and in vitro transcription–translation data collectively indicate that the lack of synthesis of the mutant proteins is caused by the immediate degradation of the expressed product by cellular proteases rather than by faulty translation of mutant LTB mRNA. Coexpression of the LTA could not rescue the degradation of LTB mutants. PMID:17911243

  7. Peracetic acid: a practical agent for sterilizing heat-labile polymeric tissue-engineering scaffolds.

    PubMed

    Yoganarasimha, Suyog; Trahan, William R; Best, Al M; Bowlin, Gary L; Kitten, Todd O; Moon, Peter C; Madurantakam, Parthasarathy A

    2014-09-01

    Advanced biomaterials and sophisticated processing technologies aim at fabricating tissue-engineering scaffolds that can predictably interact within a biological environment at the cellular level. Sterilization of such scaffolds is at the core of patient safety and is an important regulatory issue that needs to be addressed before clinical translation. In addition, it is crucial that meticulously engineered micro- and nano- structures are preserved after sterilization. Conventional sterilization methods involving heat, steam, and radiation are not compatible with engineered polymeric systems because of scaffold degradation and loss of architecture. Using electrospun scaffolds made from polycaprolactone, a low melting polymer, and employing spores of Bacillus atrophaeus as biological indicators, we compared ethylene oxide, autoclaving and 80% ethanol to a known chemical sterilant, peracetic acid (PAA), for their ability to sterilize as well as their effects on scaffold properties. PAA diluted in 20% ethanol to 1000 ppm or above sterilized electrospun scaffolds in 15 min at room temperature while maintaining nano-architecture and mechanical properties. Scaffolds treated with PAA at 5000 ppm were rendered hydrophilic, with contact angles reduced to 0°. Therefore, PAA can provide economical, rapid, and effective sterilization of heat-sensitive polymeric electrospun scaffolds that are used in tissue engineering. PMID:24341350

  8. Peracetic Acid: A Practical Agent for Sterilizing Heat-Labile Polymeric Tissue-Engineering Scaffolds

    PubMed Central

    Yoganarasimha, Suyog; Trahan, William R.; Best, Al M.; Bowlin, Gary L.; Kitten, Todd O.; Moon, Peter C.

    2014-01-01

    Advanced biomaterials and sophisticated processing technologies aim at fabricating tissue-engineering scaffolds that can predictably interact within a biological environment at the cellular level. Sterilization of such scaffolds is at the core of patient safety and is an important regulatory issue that needs to be addressed before clinical translation. In addition, it is crucial that meticulously engineered micro- and nano- structures are preserved after sterilization. Conventional sterilization methods involving heat, steam, and radiation are not compatible with engineered polymeric systems because of scaffold degradation and loss of architecture. Using electrospun scaffolds made from polycaprolactone, a low melting polymer, and employing spores of Bacillus atrophaeus as biological indicators, we compared ethylene oxide, autoclaving and 80% ethanol to a known chemical sterilant, peracetic acid (PAA), for their ability to sterilize as well as their effects on scaffold properties. PAA diluted in 20% ethanol to 1000?ppm or above sterilized electrospun scaffolds in 15?min at room temperature while maintaining nano-architecture and mechanical properties. Scaffolds treated with PAA at 5000?ppm were rendered hydrophilic, with contact angles reduced to 0. Therefore, PAA can provide economical, rapid, and effective sterilization of heat-sensitive polymeric electrospun scaffolds that are used in tissue engineering. PMID:24341350

  9. Both enzymatic and non-enzymatic properties of heat-labile enterotoxin are responsible for LT-enhanced adherence of enterotoxigenic Escherichia coli to porcine IPEC-J2 cells.

    PubMed

    Fekete, Peter Z; Mateo, Kristina S; Zhang, Weiping; Moxley, Rodney A; Kaushik, Radhey S; Francis, David H

    2013-06-28

    Previous studies in piglets indicate that heat labile enterotoxin (LT) expression enhances intestinal colonization by K88 adhesin-producing enterotoxigenic Escherichia coli (ETEC) as wild-type ETEC adhered to intestinal epithelium in substantially greater numbers than did non-toxigenic constructs. Enzymatic activity of the toxin was also shown to contribute to the adhesion of ETEC and non-ETEC bacteria to epithelial cells in culture. To further characterize the contribution of LT to host cell adhesion, a nontoxigenic, K88-producing E. coli was transformed with either the gene encoding for LT holotoxin, a catalytically-attenuated form of the toxin [LT(R192G)], or LTB subunits, and resultant changes in bacterial adherence to IPEC-J2 porcine intestinal epithelial cells were measured. Strains expressing LT holotoxin or mutants were able to adhere in significantly higher numbers to IPEC-J2 cells than was an isogenic, toxin-negative construct. LT+ strains were also able to significantly block binding of a wild-type LT+ ETEC strain to IPEC-J2 cells. Adherence of isogenic strains to IPEC-J2 cells was unaltered by cycloheximide treatment, suggesting that LT enhances ETEC adherence to IPEC-J2 cells independent of host cell protein synthesis. However, pretreating IPEC-J2 cells with LT promoted adherence of negatively charged latex beads (a surrogate for bacteria which carry a negative change), which adherence was inhibited by cycloheximide, suggesting LT may induce a change in epithelial cell membrane potential. Overall, these data suggest that LT may enhance ETEC adherence by promoting an association between LTB and epithelial cells, and by altering the surface charge of the host plasma membrane to promote non-specific adherence. PMID:23517763

  10. Inhibition of Cronobacter sakazakii by heat labile bacteriocins produced by probiotic LAB isolated from healthy infants.

    PubMed

    Awaisheh, Saddam S; Al-Nabulsi, Anas A; Osaili, Tareq M; Ibrahim, Salam; Holley, Richard

    2013-09-01

    Cronobacter sakazakii is an opportunistic pathogen that can cause bacteremia, meningitis, and necrotizing enterocolitis, most often in neonates with case-fatality rates that may reach 80%. The antimicrobial activity of lactic acid bacteria against a wide range of foodborne pathogens is well-established in different types of food products. The objective of the current study was to investigate the antibacterial activity of Lactobacillus acidophilus and L. casei isolated from feces of healthy infants against different strains of C. sakazakii in agar and a rehydrated infant milk formula (RIMF) model. The inhibition zones of C. sakazakii around L. acidophilus or L. casei ranged from 22 to 32 mm on eMan Rogosa Sharpe (MRS) agar under aerobic conditions, while a slight reduction in antibacterial activity was noted on modified MRS (0.2% glucose) under anaerobic conditions. It was observed that pH-neutralized cell-free supernatant (CFS) of L. acidophilus or L. casei was inhibitory against tested C. sakazakii strains. The inhibition zones of neutralized CFS were lower than the antibacterial activities of live cultures. The antibacterial activity of CFS was abolished when CFS from L. acidophilus or L. casei was heated at 60 or 80C for either 10 min or 2 h, or treated with trypsin or pepsin. This was considered strong evidence that the inhibition was due to the production of bacteriocins by L. casei and L. acidophilus. Both the CFS and active growing cells of L. casei and L. acidophilus were able to reduce the viability of C. sakazakii in the RIMF model. The results may extend the use of natural antimicrobials instead of conventional preservation methods to improve the safety of RIMF. PMID:23924352

  11. Levels of Expression and Immunogenicity of Attenuated Salmonella enterica Serovar Typhimurium Strains Expressing Escherichia coli Mutant Heat-Labile Enterotoxin

    PubMed Central

    Covone, M. Giuseppina; Brocchi, Marcelo; Palla, Emanuela; da Silveira, W. Dias; Rappuoli, Rino; Galeotti, Cesira L.

    1998-01-01

    The effects of heterologous gene dosage as well as Salmonella typhimurium strain variability on immune response toward both the heterologous antigen, the nontoxic mutant of the Escherichia coli heat-labile enterotoxin LTK63, and the carrier Salmonella strain have been analyzed. Effects of a single integration into the host DNA and different-copy-number episomal vectors were compared in S. typhimurium Δcya Δcrp Δasd strains of two different serotypes, UK-1 and SR-11. Expression of the enterotoxin in the different Salmonella isolates in vitro was found to vary considerably and, for the episomal vectors, to correlate with the plasmid copy number. LTK63-specific serum immunoglobulin G (IgG) and mucosal immunoglobulin A (IgA) antibodies were highest in mice immunized with the high-level-expression strain. High anti-LTK63 IgG and IgA titers were found to correspond to higher anti-Salmonella immunity, suggesting that LTK63 exerts an adjuvant effect on response to the carrier. Statistically significant differences in anti-LTK63 immune response were observed between groups of mice immunized with the attenuated Δcya Δcrp UK-1 and SR-11 derivatives producing the antigen at the same rate. These data indicate that the same attenuation in S. typhimurium strains of different genetic backgrounds can influence significantly the immune response toward the heterologous antigen. Moreover, delivery of the LTK63 enterotoxin to the immune system by attenuated S. typhimurium strains is effective only when synthesis of the antigen is very high during the initial phase of invasion, while persistence of the S. typhimurium strain in deep tissues has only marginal influence. PMID:9423862

  12. Levels of expression and immunogenicity of attenuated Salmonella enterica serovar typhimurium strains expressing Escherichia coli mutant heat-labile enterotoxin.

    PubMed

    Covone, M G; Brocchi, M; Palla, E; Dias da Silveira, W; Rappuoli, R; Galeotti, C L

    1998-01-01

    The effects of heterologous gene dosage as well as Salmonella typhimurium strain variability on immune response toward both the heterologous antigen, the nontoxic mutant of the Escherichia coli heat-labile enterotoxin LTK63, and the carrier Salmonella strain have been analyzed. Effects of a single integration into the host DNA and different-copy-number episomal vectors were compared in S. typhimurium delta cya delta crp delta asd strains of two different serotypes, UK-1 and SR-11. Expression of the enterotoxin in the different Salmonella isolates in vitro was found to vary considerably and, for the episomal vectors, to correlate with the plasmid copy number. LTK63-specific serum immunoglobulin G (IgG) and mucosal immunoglobulin A (IgA) antibodies were highest in mice immunized with the high-level-expression strain. High anti-LTK63 IgG and IgA titers were found to correspond to higher anti-Salmonella immunity, suggesting that LTK63 exerts an adjuvant effect on response to the carrier. Statistically significant differences in anti-LTK63 immune response were observed between groups of mice immunized with the attenuated delta cya delta crp UK-1 and SR-11 derivatives producing the antigen at the same rate. These data indicate that the same attenuation in S. typhimurium strains of different genetic backgrounds can influence significantly the immune response toward the heterologous antigen. Moreover, delivery of the LTK63 enterotoxin to the immune system by attenuated S. typhimurium strains is effective only when synthesis of the antigen is very high during the initial phase of invasion, while persistence of the S. typhimurium strain in deep tissues has only marginal influence. PMID:9423862

  13. Contribution of the Disulfide Bond of the A Subunit to the Action of Escherichia coli Heat-Labile Enterotoxin

    PubMed Central

    Okamoto, Keinosuke; Nomura, Tomohiko; Fujii, Yoshio; Yamanaka, Hiroyasu

    1998-01-01

    Escherichia coli heat-labile enterotoxin (LT) consists of an A subunit and five B subunits. These subunits oligomerize into an assembled holotoxin within the periplasm. Structural analysis of LT has revealed that the A subunit interacts with the B subunit through its carboxy terminus. This indicates that the carboxy-terminal portion of the protein is required for assembly of holotoxin in the periplasm. However, it is not known whether other regions of the A subunit contribute to the assembly. The A subunit constituting the holotoxin contains a disulfide bond between Cys-187 and Cys-199. It has been observed in many proteins that the intramolecular disulfide bond is deeply involved in the function and tertiary structure of the protein. We speculated that the disulfide bond of the A subunit contributes to the assembly in the periplasm, although the bond is not a structural element of the carboxy-terminal portion of the A subunit. We replaced these cysteine residues of the A subunit by oligonucleotide-directed site-specific mutagenesis and analyzed the LTs produced by cells containing the mutant LT genes. The amount of the mutant holotoxin produced was small compared with that of the wild-type strain, indicating that the disulfide bond of the A subunit contributes to the structure which functions as the site of nucleation in the assembly. A reconstitution experiment in vitro supported the notion. Subsequently, we found that the mutant A subunit constituting holotoxin is easily degraded by trypsin and that in cells incubated with mutant LTs, the lag until the intracellular cyclic AMP begins to accumulate is longer than in cells incubated with native LTs. These results might be useful for the analysis of the interaction of LT with target cells at the molecular level. PMID:9515902

  14. Trivalent heat-labile- and heat-stable-enterotoxin probe conjugated with horseradish peroxidase for detection of enterotoxigenic Escherichia coli by hybridization.

    PubMed Central

    Abe, A; Komase, K; Bangtrakulnonth, A; Ratchtrachenchat, O A; Kawahara, K; Danbara, H

    1990-01-01

    A 1,268-bp polynucleotide probe for heat-labile and heat-stable enterotoxins (LTh, STIa, STIb) was conjugated with horseradish peroxidase (HRP). The HRP-conjugated trivalent probe was applied to the detection of enterotoxigenic Escherichia coli (ETEC) by colony and stool hybridizations. The binding of the probe to its targets was assayed by the addition of HRP substrates hydrogen peroxide and luminol in the presence of an enhancer, and the chemiluminescence was recorded by exposure to X-ray film. Slot blot hybridization demonstrated that the HRP-conjugated trivalent probe specifically hybridized with the DNA isolated from ETEC strains. The trivalent probe also specifically identified bacterial colonies of ETEC that produced LTh, STIa, STIb, LTh-STIa, or LTh-STIb. Treatment of targets with sodium dodecyl sulfate and proteinase K remarkably reduced nonspecific hybridization to DNAs of non-ETEC strains. Furthermore, this probe was able to detect stool specimens seeded with 10(2) original ETEC cells per 5 mg of feces. These results suggest that the HRP-conjugated trivalent probe is a candidate for use in the clinical laboratory to detect ETEC. Images PMID:2279991

  15. Relationship between Heat-Labile Enterotoxin Secretion Capacity and Virulence in Wild Type Porcine-Origin Enterotoxigenic Escherichia coli Strains

    PubMed Central

    Wijemanne, Prageeth; Xing, Jun; Berberov, Emil M.; Marx, David B.; Francis, David H.; Moxley, Rodney A.

    2015-01-01

    Heat-labile enterotoxin (LT) is an important virulence factor secreted by some strains of enterotoxigenic Escherichia coli (ETEC). The prototypic human-origin strain H10407 secretes LT via a type II secretion system (T2SS). We sought to determine the relationship between the capacity to secrete LT and virulence in porcine-origin wild type (WT) ETEC strains. Sixteen WT ETEC strains isolated from cases of severe diarrheal disease were analyzed by GM1ganglioside enzyme-linked immunosorbent assay to measure LT concentrations in culture supernatants. All strains had detectable LT in supernatants by 2 h of culture and 1 strain, which was particularly virulent in gnotobiotic piglets (3030-2), had the highest LT secretion level all porcine-origin WT strains tested (P<0.05). The level of LT secretion (concentration in supernatants at 6-h culture) explained 92% of the variation in time-to-a-moribund-condition (R2 = 0.92, P<0.0001) in gnotobiotic piglets inoculated with either strain 3030-2, or an ETEC strain of lesser virulence (2534-86), or a non-enterotoxigenic WT strain (G58-1). All 16 porcine ETEC strains were positive by PCR analysis for the T2SS genes, gspD and gspK, and bioinformatic analysis of 4 porcine-origin strains for which complete genomic sequences were available revealed a T2SS with a high degree of homology to that of H10407. Maximum Likelihood phylogenetic trees constructed using T2SS genes gspC, gspD, gspE and homologs showed that strains 2534-86 and 3030-2 clustered together in the same clade with other porcine-origin ETEC strains in the database, UMNK88 and UMN18. Protein modeling of the ATPase gene (gspE) further revealed a direct relationship between the predicted ATP-binding capacities and LT secretion levels as follows: H10407, -8.8 kcal/mol and 199 ng/ml; 3030-2, -8.6 kcal/mol and 133 ng/ml; and 2534-86, -8.5 kcal/mol and 80 ng/ml. This study demonstrated a direct relationship between predicted ATP-binding capacity of GspE and LT secretion, and between the latter and virulence. PMID:25768732

  16. Treatment of PCR products with exonuclease I and heat-labile alkaline phosphatase improves the visibility of combined bisulfite restriction analysis

    SciTech Connect

    Watanabe, Kousuke; Emoto, Noriko; Sunohara, Mitsuhiro; Kawakami, Masanori; Kage, Hidenori; Nagase, Takahide; Ohishi, Nobuya; Takai, Daiya

    2010-08-27

    Research highlights: {yields} Incubating PCR products at a high temperature causes smears in gel electrophoresis. {yields} Smears interfere with the interpretation of methylation analysis using COBRA. {yields} Treatment with exonuclease I and heat-labile alkaline phosphatase eliminates smears. {yields} The elimination of smears improves the visibility of COBRA. -- Abstract: DNA methylation plays a vital role in the regulation of gene expression. Abnormal promoter hypermethylation is an important mechanism of inactivating tumor suppressor genes in human cancers. Combined bisulfite restriction analysis (COBRA) is a widely used method for identifying the DNA methylation of specific CpG sites. Here, we report that exonuclease I and heat-labile alkaline phosphatase can be used for PCR purification for COBRA, improving the visibility of gel electrophoresis after restriction digestion. This improvement is observed when restriction digestion is performed at a high temperature, such as 60 {sup o}C or 65 {sup o}C, with BstUI and TaqI, respectively. This simple method can be applied instead of DNA purification using spin columns or phenol/chloroform extraction. It can also be applied to other situations when PCR products are digested by thermophile-derived restriction enzymes, such as PCR restriction fragment length polymorphism (RFLP) analysis.

  17. Mutants of the Escherichia coli heat-labile enterotoxin with reduced ADP-ribosylation activity or no activity retain the immunogenic properties of the native holotoxin.

    PubMed Central

    de Haan, L; Verweij, W R; Feil, I K; Lijnema, T H; Hol, W G; Agsteribbe, E; Wilschut, J

    1996-01-01

    The Escherichia coli heat-labile enterotoxin (LT) is a potent inducer of mucosal immune responses. In a previous study (L. DeHaan, W. R. Verweij, M. Holtrop, E. Agsteribbe, and J. Wilschut, Vaccine 14:620-626, 1996), we have shown that efficient induction of an LTB-specific mucosal immune response by LT requires the presence of the LTA chain, suggesting a possible role of the enzymatic activity of LTA in the induction of these responses. In the present study, we generated LT mutants with altered ADP-ribosylation activities and evaluated their immunogenicity upon intranasal administration to mice. The results demonstrate that the mucosal immunogenicity of LT is not dependent on its ADP-ribosylation activity. PMID:8945598

  18. A pain-inducing centipede toxin targets the heat activation machinery of nociceptor TRPV1

    NASA Astrophysics Data System (ADS)

    Yang, Shilong; Yang, Fan; Wei, Ningning; Hong, Jing; Li, Bowen; Luo, Lei; Rong, Mingqiang; Yarov-Yarovoy, Vladimir; Zheng, Jie; Wang, Kewei; Lai, Ren

    2015-09-01

    The capsaicin receptor TRPV1 ion channel is a polymodal nociceptor that responds to heat with exquisite sensitivity through an unknown mechanism. Here we report the identification of a novel toxin, RhTx, from the venom of the Chinese red-headed centipede that potently activates TRPV1 to produce excruciating pain. RhTx is a 27-amino-acid small peptide that forms a compact polarized molecule with very rapid binding kinetics and high affinity for TRPV1. We show that RhTx targets the channel's heat activation machinery to cause powerful heat activation at body temperature. The RhTx-TRPV1 interaction is mediated by the toxin's highly charged C terminus, which associates tightly to the charge-rich outer pore region of the channel where it can directly interact with the pore helix and turret. These findings demonstrate that RhTx binding to the outer pore can induce TRPV1 heat activation, therefore providing crucial new structural information on the heat activation machinery.

  19. A pain-inducing centipede toxin targets the heat activation machinery of nociceptor TRPV1

    PubMed Central

    Yang, Shilong; Yang, Fan; Wei, Ningning; Hong, Jing; Li, Bowen; Luo, Lei; Rong, Mingqiang; Yarov-Yarovoy, Vladimir; Zheng, Jie; Wang, KeWei; Lai, Ren

    2015-01-01

    The capsaicin receptor TRPV1 ion channel is a polymodal nociceptor that responds to heat with exquisite sensitivity through an unknown mechanism. Here we report the identification of a novel toxin, RhTx, from the venom of the Chinese red-headed centipede that potently activates TRPV1 to produce excruciating pain. RhTx is a 27-amino-acid small peptide that forms a compact polarized molecule with very rapid binding kinetics and high affinity for TRPV1. We show that RhTx targets the channel's heat activation machinery to cause powerful heat activation at body temperature. The RhTx–TRPV1 interaction is mediated by the toxin's highly charged C terminus, which associates tightly to the charge-rich outer pore region of the channel where it can directly interact with the pore helix and turret. These findings demonstrate that RhTx binding to the outer pore can induce TRPV1 heat activation, therefore providing crucial new structural information on the heat activation machinery. PMID:26420335

  20. A pain-inducing centipede toxin targets the heat activation machinery of nociceptor TRPV1.

    PubMed

    Yang, Shilong; Yang, Fan; Wei, Ningning; Hong, Jing; Li, Bowen; Luo, Lei; Rong, Mingqiang; Yarov-Yarovoy, Vladimir; Zheng, Jie; Wang, KeWei; Lai, Ren

    2015-01-01

    The capsaicin receptor TRPV1 ion channel is a polymodal nociceptor that responds to heat with exquisite sensitivity through an unknown mechanism. Here we report the identification of a novel toxin, RhTx, from the venom of the Chinese red-headed centipede that potently activates TRPV1 to produce excruciating pain. RhTx is a 27-amino-acid small peptide that forms a compact polarized molecule with very rapid binding kinetics and high affinity for TRPV1. We show that RhTx targets the channel's heat activation machinery to cause powerful heat activation at body temperature. The RhTx-TRPV1 interaction is mediated by the toxin's highly charged C terminus, which associates tightly to the charge-rich outer pore region of the channel where it can directly interact with the pore helix and turret. These findings demonstrate that RhTx binding to the outer pore can induce TRPV1 heat activation, therefore providing crucial new structural information on the heat activation machinery. PMID:26420335

  1. Incorporation of membrane-anchored flagellin or Escherichia coli heat-labile enterotoxin B subunit enhances the immunogenicity of rabies virus-like particles in mice and dogs.

    PubMed

    Qi, Yinglin; Kang, Hongtao; Zheng, Xuexing; Wang, Hualei; Gao, Yuwei; Yang, Songtao; Xia, Xianzhu

    2015-01-01

    Rabies remains an important worldwide public health threat, so safe, effective, and affordable vaccines are still being sought. Virus-like particle-based vaccines targeting various viral pathogens have been successfully produced, licensed, and commercialized. Here, we designed and constructed two chimeric rabies virus-like particles (cRVLPs) containing rabies virus (RABV) glycoprotein (G), matrix (M) protein, and membrane-anchored flagellin (EVLP-F) or Escherichia coli heat-labile enterotoxin B subunit (EVLP-L) as molecular adjuvants to enhance the immune response against rabies. The immunogenicity and potential of cRVLPs as novel rabies vaccine were evaluated by intramuscular vaccination in mouse and dog models. Mouse studies demonstrated that both EVLP-F and EVLP-L induced faster and larger virus-neutralizing antibodies (VNAs) responses and elicited greater numbers of CD4(+) and CD8(+) T cells secreting IFN-? or IL-4 compared with a standard rabies VLP (sRVLP) containing only G and M. Moreover, cRVLPs recruited and/or activated more B cells and dendritic cells in inguinal lymph nodes. EVLP-F induced a strong, specific IgG2a response but not an IgG1 response, suggesting the activation of Th1 class immunity; in contrast, Th2 class immunity was observed with EVLP-L. The significantly enhanced humoral and cellular immune responses induced by cRVLPs provided complete protection against lethal challenge with RABV. Most importantly, dogs vaccinated with EVLP-F or EVLP-L exhibited increased VNA titers in sera and enhanced IFN-? and IL-4 secretion from peripheral blood mononuclear cells. Taken together, these results illustrate that when incorporated into sRVLP, membrane-anchored flagellin, and heat-labile enterotoxin B subunit possess strong adjuvant activity. EVLP-F and EVLP-L induce significantly enhanced RABV-specific humoral and cellular immune responses in both mouse and dog. Therefore, these cRVLPs may be developed as safe and more efficacious rabies vaccine candidate for animals. PMID:25784906

  2. Incorporation of membrane-anchored flagellin or Escherichia coli heat-labile enterotoxin B subunit enhances the immunogenicity of rabies virus-like particles in mice and dogs

    PubMed Central

    Qi, Yinglin; Kang, Hongtao; Zheng, Xuexing; Wang, Hualei; Gao, Yuwei; Yang, Songtao; Xia, Xianzhu

    2015-01-01

    Rabies remains an important worldwide public health threat, so safe, effective, and affordable vaccines are still being sought. Virus-like particle-based vaccines targeting various viral pathogens have been successfully produced, licensed, and commercialized. Here, we designed and constructed two chimeric rabies virus-like particles (cRVLPs) containing rabies virus (RABV) glycoprotein (G), matrix (M) protein, and membrane-anchored flagellin (EVLP-F) or Escherichia coli heat-labile enterotoxin B subunit (EVLP-L) as molecular adjuvants to enhance the immune response against rabies. The immunogenicity and potential of cRVLPs as novel rabies vaccine were evaluated by intramuscular vaccination in mouse and dog models. Mouse studies demonstrated that both EVLP-F and EVLP-L induced faster and larger virus-neutralizing antibodies (VNAs) responses and elicited greater numbers of CD4+ and CD8+ T cells secreting IFN-γ or IL-4 compared with a standard rabies VLP (sRVLP) containing only G and M. Moreover, cRVLPs recruited and/or activated more B cells and dendritic cells in inguinal lymph nodes. EVLP-F induced a strong, specific IgG2a response but not an IgG1 response, suggesting the activation of Th1 class immunity; in contrast, Th2 class immunity was observed with EVLP-L. The significantly enhanced humoral and cellular immune responses induced by cRVLPs provided complete protection against lethal challenge with RABV. Most importantly, dogs vaccinated with EVLP-F or EVLP-L exhibited increased VNA titers in sera and enhanced IFN-γ and IL-4 secretion from peripheral blood mononuclear cells. Taken together, these results illustrate that when incorporated into sRVLP, membrane-anchored flagellin, and heat-labile enterotoxin B subunit possess strong adjuvant activity. EVLP-F and EVLP-L induce significantly enhanced RABV-specific humoral and cellular immune responses in both mouse and dog. Therefore, these cRVLPs may be developed as safe and more efficacious rabies vaccine candidate for animals. PMID:25784906

  3. Bacteria and their toxins tamed for immunotherapy.

    PubMed

    Adkins, Irena; Holubova, Jana; Kosova, Martina; Sadilkova, Lenka

    2012-06-01

    Bacterial toxins share the ability to enter host cells to target various intracellular proteins and to modulate host immune responses. Over the last 20 years, toxins and their mutated variants, as well as live attenuated bacteria, have been exploited for vaccination and immunotherapy of various infectious, malignant and autoimmune diseases. The ability of Bordetella pertussis adenylate cyclase toxin to translocate its adenylate cyclase domain across the host cell membrane, as well as the pathways of intracellular trafficking of Bacillus anthracis lethal and edema toxins, Shigella dysenteriae shiga toxin or Escherichia coli shiga-like toxin, have been repeatedly exploited for the delivery of antigenic epitopes into host cells and for stimulation of antigen-specific T cell responses. Similarly, E. coli α-hemolysin, or effector proteins of Yersinia and Salmonella secreted by the type III secretion systems, were used to facilitate the delivery of fused heterologous proteins or peptides for antigenic presentation. Vibrio cholerae cholera toxin, E. coli heat-labile enterotoxin, B. pertussis pertussis toxin or the Cry1A protein of Bacillus thuringiensis have shown a great potential to act as adjuvants and to stimulate mucosal as well as systemic immune responses. The immunotherapeutic potential of some toxins, like Clostridium perfringens perfringolysin O, Streptococcus intermedius intermedilysin, or Streptococcus pneumoniae pneumolysin needs to be evaluated further. The Bordetella adenylate cyclase toxoid used as a vaccine delivery tool, or Corynebacterium diphtheriae diphtheria toxin and Pseudomonas aeruginosa exotoxin A-based immunotoxins, are currently in various phases of clinical trials for cancer immunotherapy, as are some antigen-delivering Salmonella and Listeria monocytogenes strains. PMID:22339216

  4. Local and systemic immune responses induced by a recombinant chimeric protein containing Mycoplasma hyopneumoniae antigens fused to the B subunit of Escherichia coli heat-labile enterotoxin LTB.

    PubMed

    Marchioro, Silvana Beutinger; Fisch, Andressa; Gomes, Charles K; Jorge, Sérgio; Galli, Vanessa; Haesebrouck, Freddy; Maes, Dominiek; Dellagostin, Odir; Conceição, Fabricio R

    2014-09-17

    A multi-antigen chimera composed of three antigens of Mycoplasma hyopneumoniae (R1, P42, and NrdF) and the mucosal adjuvant Escherichia coli heat-labile enterotoxin B subunit (LTB) was constructed, and its antigenic and immunogenic properties were evaluated in mice and pigs. In addition, we compared the effect of the fusion and co-administration of these proteins in mice. Antibodies against each subunit recognized the chimeric protein. Intranasal and intramuscular immunization of mice with the chimeric protein significantly increased IgG and IgA levels in the serum and tracheobronchial lavages, respectively, against some of the antigens present in the chimeric. Swine immunized with the chimeric protein developed an immune response against all M. hyopneumoniae antigens present in the fusion with a statistically significant difference (P<0.05). The adjuvant rLTB enhanced the immune response in both fused and co-administered antigens; however, better results were obtained with the chimeric protein. This multi-antigen is a promising vaccine candidate that may help control M. hyopneumoniae infection. PMID:25091529

  5. Influence of heat-labile serum components in the presence of OmpA on the outer membrane of Salmonella gallinarum.

    PubMed

    Vega-Manriquez, X; Huerta-Ascencio, L; Martínez-Gómez, D; López-Vidal, Y; Verdugo-Rodríguez, A

    2016-03-01

    Salmonella gallinarum is the causative agent of fowl typhoid. Being a Gram-negative bacteria, its outer membrane proteins (OMP) can be regulated by different microenvironments. S. gallinarum was cultured under the following conditions: nutrient broth (NB), NB supplemented with serum from specific pathogen-free birds (NBS) and NB with serum incubated at 56 °C prior to incubation with the bacteria (NBSD); OMP were subsequently extracted. Several changes were observed in the apparent expression of OMP, mainly a decrease in an OMP with a size of 30 kDa, approximately, under the NBS condition. In contrast, the same event was not observed in NB and NBSD when using one- and two-dimensional polyacrylamide gels (SDS-PAGE). Using the OMP with a size of 30 kDa, approximately, as antigen in indirect ELISA, we were able to differentiate serum from healthy and vaccinated birds, as well as birds infected with S. gallinarum and S. enteritidis. The amino-terminal of this protein was sequenced, showing 100 % identity with OmpA of S. typhimurium. Subsequently, we designed primers to amplify the gene by PCR. The partial sequence of the amplified gene showed 100 % identity with OmpA of S. gallinarum. (1) Heat-labile serum components influence the presence of OmpA in the OM of S. gallinarum; (2) by the way of ELISA, OmpA allows to specifically differentiate healthy from diseased birds. PMID:26597854

  6. Potent immunogenicity of the B subunits of Escherichia coli heat-labile enterotoxin: receptor binding is essential and induces differential modulation of lymphocyte subsets.

    PubMed Central

    Nashar, T O; Webb, H M; Eaglestone, S; Williams, N A; Hirst, T R

    1996-01-01

    The importance of receptor binding in the potent immunogenicity of Escherichia coli heat-labile enterotoxin B subunit (EtxB) was tested by comparing its immunogical properties with those of a receptor binding mutant, EtxB(G33D). Subcutaneous immunization of EtxB(G33D) resulted in 160-fold reduction in antibody titer compared with wild-type EtxB, whereas its oral delivery failed to provoke any detectable secretory or serum anti-B subunit responses. Moreover, the two proteins induced strikingly different effects on lymphocyte cultures in vitro. EtxB, in comparison with EtxB(G33D), caused an increase in the proportion of B cells, many of which were activated (CD25+); the complete depletion of CD8+ T cells; an increase in the activation of CD4+ T cells; and an increase in interleukin 2 and a decrease in interferon gamma. These data indicate that EtxB exerts profound effects on immune cells, suggesting that its potent immunogenicity is dependent not only on efficient receptor-mediated uptake, but also on direct receptor-mediated immunomodulation of lymphocyte subsets. Images Fig. 1 Fig. 2 Fig. 4 Fig. 5 PMID:8552610

  7. Alterations at the carboxyl terminus change assembly and secretion properties of the B subunit of Escherichia coli heat-labile enterotoxin.

    PubMed Central

    Sandkvist, M; Hirst, T R; Bagdasarian, M

    1987-01-01

    The gene encoding the B subunit of heat-labile enterotoxin (etxB) was mutated at its 3' end by targeted addition of random nucleotide sequences. Gene products from five mutated etxB genes, all of which were shown to encode B subunits with short carboxy-terminal amino acid extensions, were analyzed with respect to a range of functional and structural properties. One class of altered B subunits, exemplified by EtxB124 and EtxB138, which both have seven extra amino acid residues, were found to be specifically defective in their ability to stably associate with A subunits and form holotoxin. Other altered B subunits were less subtlely affected by extensions at their C termini and were, in addition to their failure to associate with A subunits, unable to translocate into the periplasm of Escherichia coli, to pentamerize, or to bind to GM1 ganglioside. This suggests that the carboxy-terminal domain of EtxB mediates A subunit-B subunit interaction. Images PMID:2820934

  8. Analysis of Heat-Labile Sites Generated by Reactions of Depleted Uranium and Ascorbate in Plasmid DNA

    PubMed Central

    Wilson, Janice; Young, Ashley; Civitello, Edgar R.

    2013-01-01

    The goal of this study was to characterize how depleted uranium (DU) causes DNA damage. Procedures were developed to assess the ability of organic and inorganic DNA adducts to convert to single strand breaks (SSB) in pBR322 plasmid DNA in the presence of heat or piperidine. DNA adducts formed by methyl methanesulfonate (MMS), cis-platin (cis-Pt), and chromic chloride were compared to those formed by reaction of uranyl acetate (UA) and ascorbate (Asc). Uranyl ion in the presence of Asc produced U-DNA adducts that converted to SSB upon heating. Piperidine, which acted on DNA methylated by MMS to convert methyl-DNA adducts to SSB, served in the opposite fashion with U-DNA adducts by decreasing SSB. The observation that piperidine also decreased the gel shift for metal-DNA adducts formed by monofunctional cis-Pt and chromic chloride was interpreted to suggest that piperidine served to remove U-DNA adducts. Radical scavengers did not affect formation of U-induced SSB, suggesting that SSB arose from the presence of U-DNA adducts and not from free radicals. A model is proposed to predict how U-DNA adducts may serve as initial lesions that convert to SSB or AP sites. Results suggest that DU can act as a chemical genotoxin that does not require radiation for its mode of action. Characterizing the DNA lesions formed by DU is necessary to assess the relative importance of different DNA lesions in the formation of DU-induced mutations. Understanding mechanisms of formation of DU-induced mutations may contribute to identification of biomarkers of DU exposures in humans. PMID:24218036

  9. Immunization with a Double-Mutant (R192G/L211A) of the Heat-Labile Enterotoxin of Escherichia coli Offers Partial Protection against Campylobacter jejuni in an Adult Mouse Intestinal Colonization Model

    PubMed Central

    Albert, M. John; Haridas, Shilpa; Ebenezer, Mathew; Raghupathy, Raj; Khan, Islam

    2015-01-01

    We have previously shown that antibodies to cholera toxin (CT) reacted with the major outer membrane proteins (MOMPs) from Campylobacter jejuni strains on Western blot. Further, oral immunization with CT significantly protected against challenge with C. jejuni in an adult mouse colonization model of infection. CT and the heat-labile enterotoxin (LT) of enterotoxigenic Escherichia coli are structurally and functionally related. LT and its mutants including the double-mutant LT (R192G/L211A) (dmLT), are powerful mucosal adjuvants. Unlike LT which is reactogenic, dmLT has been shown to be safe for human use. In the current study, we determined whether rabbit anti-dmLT antibodies reacted with MOMPs from C. jejuni strains and whether immunization with dmLT would afford protection against C. jejuni. On Western blot, the MOMPs from C. jejuni 48 (Penner serotype O:19), C. jejuni 75 (O:3) and C. jejuni 111 (O:1,44) were probed with rabbit antibodies to dmLT or LT-E112K (a non-toxic LT mutant), which showed a lack of reaction. Adult BALB/c mice were orally immunized with dmLT and orally challenged with C. jejuni 48 or 111. Protection from colonization with the challenge bacteria was studied by enumerating Campylobacter colonies in feces daily for 9 days. Vaccination produced robust serum and stool antibody responses to dmLT and no antibody responses to C. jejuni MOMP. Vaccinated mice showed reduced colonization and excretion of both challenge strains compared to control mice. However, the differences were not statistically significant. The protective efficacy of the dmLT vaccine varied from 9.1% to 54.5%. The lack of cross-reaction between the MOMP and dmLT suggests that protection is not mediated by cross-reacting antibodies, but may be due to activation of innate immunity. As dmLT is safe for humans, it could be incorporated into a C. jejuni vaccine to enhance its efficacy. PMID:26540197

  10. Salmonella enterica Serovar Enteritidis Ghosts Carrying the Escherichia coli Heat-Labile Enterotoxin B Subunit Are Capable of Inducing Enhanced Protective Immune Responses

    PubMed Central

    Jawale, Chetan V.

    2014-01-01

    The Escherichia coli heat-labile enterotoxin B subunit (LTB) is a potent vaccine adjuvant. Salmonella enterica serovar Enteritidis ghosts carrying LTB (S. Enteritidis-LTB ghosts) were genetically constructed using a novel plasmid, pJHL187-LTB, designed for the coexpression of the LTB and E lysis proteins. S. Enteritidis-LTB ghosts were characterized using scanning electron microscopy to visualize their transmembrane tunnel structures. The expression of LTB in S. Enteritidis-LTB ghost preparations was confirmed by immunoblot and enzyme-linked immunosorbent assays. The parenteral adjuvant activity of LTB was demonstrated by immunizing chickens with either S. Enteritidis-LTB ghosts or S. Enteritidis ghosts. Chickens were intramuscularly primed at 5 weeks of age and subsequently boosted at 8 weeks of age. In total, 60 chickens were equally divided into three groups (n = 20 for each): group A, nonvaccinated control; group B, immunized with S. Enteritidis-LTB ghosts; and group C, immunized with S. Enteritidis ghosts. Compared with the nonimmunized chickens (group A), the immunized chickens (groups B and C) exhibited increased titers of plasma IgG and intestinal secretory IgA antibodies. The CD3+ CD4+ subpopulation of T cells was also significantly increased in both immunized groups. Among the immunized chickens, those in group B exhibited significantly increased titers of specific plasma IgG and intestinal secretory IgA (sIgA) antibodies compared with those in group C, indicating the immunomodulatory effects of the LTB adjuvant. Furthermore, both immunized groups exhibited decreased bacterial loads in their feces and internal organs. These results indicate that parenteral immunization with S. Enteritidis-LTB ghosts can stimulate superior induction of systemic and mucosal immune responses compared to immunization with S. Enteritidis ghosts alone, thus conferring efficient protection against salmonellosis. PMID:24671556

  11. Decreased Heat-Labile Opsonic Activity and Complement Levels Associated with Evidence of C3 Breakdown Products in Infected Pleural Effusions

    PubMed Central

    Lew, Pablo D.; Zubler, Rudolf; Vaudaux, Pierre; Farquet, Jean J.; Waldvogel, Francis A.; Lambert, Paul-Henri

    1979-01-01

    Heat-labile opsonic activity was measured simultaneously in serum and pleural fluid of patients with transudates, infectious exudates (with positive or negative bacterial culture) and neoplastic exudates, using two different complement-dependent phagocytic tests: the killing of Staphylococcus aureus Wood 46 variant strain (K50 opsonic titers) and the assessment of ingestion rate of endotoxin-coated paraffin particles (Oil Red 0 uptake test). K50 opsonic titers were lower in culture-positive pleural effusions as compared to culture-negative (P < 0.002) or neoplastic effusions (P < 0.002). These results were corroborated by the Oil Red 0 uptake test. The data obtained with the two assays showed a significant correlation (P < 0.001). The hemolytic activity of complement (CH50) as well as the levels of C3 breakdown product, C3d, were measured in the same sera and pleural fluid samples and in an additional group of patients with pleural effusions of the same etiology. Effusions with positive cultures showed lower CH50 values (P < 0.01) and higher C3d values (P < 0.05) when compared to culture-negative pleural fluids. Finally, evidence for immune complexes in pleural effusions and sera was looked for by determination of Clq binding activity. Levels were higher in culture-positive effusions when compared to culture-negative fluids (P = 0.005). K50 opsonic titers showed a positive correlation with CH50 values (P < 0.001) for all fluids tested. Similarly Clq binding activity correlated with C3d levels in effusions of infectious origin (P = 0.05). Recovery experiments using the various bacterial species isolated from culture-positive pleural effusions showed evidence of complement inactivation upon incubation with pooled sera at concentrations of 107-108 microorganisms/ml. These results indicate that one important reason for bacterial persistence in empyema may be decreased opsonization secondary to local consumption of complement. Images PMID:429556

  12. Human papillomavirus virus-like particles are efficient oral immunogens when coadministered with Escherichia coli heat-labile enterotoxin mutant R192G or CpG DNA.

    PubMed

    Gerber, S; Lane, C; Brown, D M; Lord, E; DiLorenzo, M; Clements, J D; Rybicki, E; Williamson, A L; Rose, R C

    2001-05-01

    Certain human papillomaviruses (HPVs) cause most cervical cancer, which remains a significant source of morbidity and mortality among women worldwide. HPV recombinant virus-like particles (VLPs) are promising vaccine candidates for controlling anogenital HPV disease and are now being evaluated as a parenteral vaccine modality in human subjects. Vaccines formulated for injection generally are more costly, more difficult to administer, and less acceptable to recipients than are mucosally administered vaccines. Since oral delivery represents an attractive alternative to parenteral injection for large-scale human vaccination, the oral immunogenicity of HPV type 11 (HPV-11) VLPs in mice was previously investigated; it was found that a modest systemic neutralizing antibody response was induced (R. C. Rose, C. Lane, S. Wilson, J. A. Suzich, E. Rybicki, and A. L. Williamson, Vaccine 17:2129-2135, 1999). Here we examine whether VLPs of other genotypes may also be immunogenic when administered orally and whether mucosal adjuvants can be used to enhance VLP oral immunogenicity. We show that HPV-16 and HPV-18 VLPs are immunogenic when administered orally and that oral coadministration of these antigens with Escherichia coli heat-labile enterotoxin (LT) mutant R192G (LT R192G) or CpG DNA can significantly improve anti-VLP humoral responses in peripheral blood and in genital mucosal secretions. Our results also suggest that LT R192G may be superior to CpG DNA in this ability. These findings support the concept of oral immunization against anogenital HPV disease and suggest that clinical studies involving this approach may be warranted. PMID:11312347

  13. Mucosal Adjuvanticity and Immunogenicity of LTR72, a Novel Mutant of Escherichia coli Heat-labile Enterotoxin with Partial Knockout of ADP-ribosyltransferase Activity

    PubMed Central

    Giuliani, Marzia Monica; Del Giudice, Giuseppe; Giannelli, Valentina; Dougan, Gordon; Douce, Gill; Rappuoli, Rino; Pizza, Mariagrazia

    1998-01-01

    Heat-labile Escherichia coli enterotoxin (LT) has the innate property of being a strong mucosal immunogen and adjuvant. In the attempt to reduce toxicity and maintain the useful immunological properties, several LT mutants have been produced. Some of these are promising mucosal adjuvants. However, so far, only those that were still toxic maintained full adjuvanticity. In this paper we describe a novel LT mutant with greatly reduced toxicity that maintains most of the adjuvanticity. The new mutant (LTR72), that contains a substitution Ala → Arg in position 72 of the A subunit, showed only 0.6% of the LT enzymatic activity, was 100,000-fold less toxic than wild-type LT in Y1 cells in vitro, and was at least 20 times less effective than wild-type LT in the rabbit ileal loop assay in vivo. At a dose of 1 μg, LTR72 exhibited a mucosal adjuvanticity, similar to that observed with wild-type LT, better than that induced by the nontoxic, enzymatically inactive LTK63 mutant, and much greater than that of the recombinant B subunit. This trend was consistent for both the amounts and kinetics of the antibody induced, and priming of antigen-specific T lymphocytes. The data suggest that the innate high adjuvanticity of LT derives from the independent contribution of the nontoxic AB complex and the enzymatic activity. LTR72 optimizes the use of both properties: the enzymatic activity for which traces are enough, and the nontoxic AB complex, the effect of which is dose dependent. In fact, in dose–response experiments in mice, 20 μg of LTR72 were a stronger mucosal adjuvant than wild-type LT. This suggests that LTR72 may be an excellent candidate to be tested in clinical trials. PMID:9529328

  14. Thermal Inactivation of Type E Botulinum Toxin1

    PubMed Central

    Licciardello, Joseph J.; Nickerson, John T. R.; Ribich, Crystal A.; Goldblith, Samuel A.

    1967-01-01

    The theoretical required cooking times for inactivation of type E Clostridium botulinum toxin (5,000 ld50 mouse units per 0.5 ml) in haddock fillets of various sizes were calculated by graphical integration of the toxin inactivation rate and heat penetration data. The results indicated that normal cooking procedures should suffice to inactivate this amount of toxin. This conclusion was substantiated by the following additional experimental observations which revealed that the original experiments had been conducted under conservative conditions. First, maximal heat stability of the toxin was found to occur at about pH 5.5, with decreasing resistance upon increasing pH. The theoretical cooking times were based on destruction of the toxin at pH 6.7. The pH of radio-pasteurized inoculated haddock, when toxin production had occurred, was on the alkaline side, at which condition the toxin is heat-labile. Second, when spoilage was discernible in radio-pasteurized inoculated haddock, the toxin titer was low, about 50 ld50 mouse units per 0.5 ml. Third, the toxin was adequately inactivated in toxic fillets after deep-fat frying for 3 min at 375 F (190.6 C) or after pan frying for 5 min per side at 400 F (204.4 C). Fourth, in this study, residual toxin activity was assayed by intraperitoneal injection of mice. It was shown that the oral toxic dose was 50 to 100 times greater than the intraperitoneal toxic dose. PMID:5339838

  15. Marine Neurotoxins: Envenomations and Contact Toxins.

    PubMed

    Watters, Michael R.; Stommel, Elijah W.

    2004-03-01

    Familiarity with the appearance and habitat of venomous sea creatures, the location of their stinging apparatus, and surveillance of population concentrations within recreational waters are essential in avoiding envenomations. Compared with the thermo-stable low molecular weighted ingestible seafood toxins, venomous toxins are often large molecular weight proteins and many are heat labile, which provides opportunity for therapeutic intervention. Heat therapy may denature the toxins, and provide immediate relief of pain in coelenterate and venomous fish envenomations. Injections of local anesthetic agents may also be used. First aid measures at the seashore may limit the spread of venom, and include immobilization of the affected sites, compression bandaging, and venous-lymphatic occlusive bandages. Measures to limit continued envenomation by attached stinging cells include topical vinegar for jellyfish tentacles and irrigation with debridment for spines of venomous fish. Antivenins are of limited availability and may be used for envenomations with sea snakes, Chironex box jellyfish, and some venomous fish. Sea snakes bites may be treated with antivenin from land snakes or with hemodialysis when antivenin is not available. Neuromuscular paralysis occurs with bites by sea snakes, cone snails, blue octopuses, and some jellyfish. Supportive treatment includes attention to cardiopulmonary resuscitation and intubation. Exposure to Pfeisteria may result in cognitive and behavioral abnormalities. Treatment with cholestyramine may be helpful in binding the toxin and improve recovery. PMID:14759344

  16. Shiga toxin Stx2 is heat-stable and not inactivated by pasteurization

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Shiga toxins are expressed by Escherichia coli (STEC) and are associated with food-borne diseases. Pasteurization is used to retard microbial growth in milk, and an open question is whether milk pasteurization inactivates shiga toxins. To answer this question we measure shiga toxin’s inhibition effe...

  17. Analysis and application of a neutralizing linear epitope on liable toxin B of enterotoxin Escherichia coli.

    PubMed

    Guan, Weikun; Liu, Wenxin; Bao, Jun; Li, Jinping; Yuan, Chaowen; Tang, Jie; Shi, Dongfang

    2015-07-01

    Heat-labile enterotoxin (LT) of enterotoxigenic Escherichia coli (ETEC) is one of the major virulence factors for causing diarrhea in piglets, and LT is a strong immunogen. Thus, LT represents an important target for development of vaccines and diagnostic tests. In this study, bioinformatic tools were used to predict six antigenic B cell epitopes in the B subunit of LT protein (LTB) of ETEC strains. Then, seven antigenic B cell epitopes of LTB were identified by polyclonal antisera (polyclonal antibody (PAb)) using a set of LTB-derived peptides expressed as maltose-binding protein (MBP) fusion protein. In addition, one LTB-specific monoclonal antibody (MAb) was generated and defined its corresponding epitope as mentioned above. This MAb was able to specifically bind with native LT toxin and has no cross-reaction with LT-II (type II heat-labile enterotoxin), Stx1 (Shiga toxin I), Stx2 (Shiga toxin II), STa (heat-stable enterotoxin I), and STb (heat-stable enterotoxin II) toxins. Further, this MAb was able to interrupt LT toxin specific binding to GM1 receptor, indicating that the corresponding epitope is the specific binding region to GM1 receptor. Moreover, in vitro and in vivo assay showed that the MAb was able to neutralize the native LT toxin. Diarrheal suckling pigs challenged with LT-positive ETEC strain recovered when an enema with this purified MAb was administered. This study will provide the foundation for further studies about the interaction between LT toxin and GM1 receptor and about the developing of epitope-based vaccines and specific therapeutic agent. PMID:25794873

  18. The chromosomal nature of LT-II enterotoxins solved: a lambdoid prophage encodes both LT-II and one of two novel pertussis-toxin-like toxin family members in type II enterotoxigenic Escherichia coli.

    PubMed

    Jobling, Michael G

    2016-04-01

    Heat-labile enterotoxins (LT) of enterotoxigenic Escherichia coli (ETEC) are structurally and functionally related to cholera toxin (CT). LT-I toxins are plasmid-encoded and flanked by IS elements, while LT-II toxins of type II ETEC are chromosomally encoded with flanking genes that appear phage related. Here, I determined the complete genomic sequence of the locus for the LT-IIa type strain SA53, and show that the LT-IIa genes are encoded by a 51 239 bp lambdoid prophage integrated at the rac locus, the site of a defective prophage in E. coli K12 strains. Of 50 LT-IIa and LT-IIc, 46 prophages also encode one member of two novel two-gene ADP-ribosyltransferase toxin families that are both related to pertussis toxin, which I named eplBA or ealAB, respectively. The eplBA and ealAB genes are syntenic with the Shiga toxin loci in their lambdoid prophages of the enteric pathogen enterohemorrhagic E. coli. These novel AB5 toxins show pertussis-toxin-like activity on tissue culture cells, and like pertussis toxin bind to sialic acid containing glycoprotein ligands. Type II ETEC are the first mucosal pathogens known to simultaneously produce two ADP-ribosylating toxins predicted to act on and modulate activity of both stimulatory and inhibitory alpha subunits of host cell heterotrimeric G-proteins. PMID:26755534

  19. Characteristics of the labile neurotoxin associated with nervous coccidiosis.

    PubMed Central

    Isler, C M; Bellamy, J E; Wobeser, G A

    1987-01-01

    Reported are the results of preliminary attempts to characterize the molecular weight, heat sensitivity and other features of a labile neurotoxin identified in the serum of calves exhibiting neurological signs in association with coccidial enteritis. The labile neurotoxin activity is heat labile (60 degrees C for 30 min) and is lost upon exposure to acidic pH (5.5) and cysteine (1.75 g/100 mL serum). Activity can be recovered from the precipitate of a 30% wt/vol solution of (NH4)2SO4 in serum. Ultrafiltration trials suggest that labile neurotoxin activity may be linked to a molecule of over 300,000 MW. PMID:2955866

  20. Heat-induced transcription of diphtheria toxin A or its variants, CRM176 and CRM197: implications for pancreatic cancer gene therapy.

    PubMed

    Fogar, P; Navaglia, F; Basso, D; Zambon, C-F; Moserle, L; Indraccolo, S; Stranges, A; Greco, E; Fadi, E; Padoan, A; Pantano, G; Sanzari, M C; Pedrazzoli, S; Montecucco, C; Plebani, M

    2010-01-01

    Vectors combining the heat shock proteins (HSPs) promoter with the catalytic subunit A of the diphtheria toxin (DTA) or its variants, cross-reacting material (CRM) 176 and 197, were engineered to investigate the effect of bacterial toxins on pancreatic cancer (PC) cells. Three heat-inducible enhanced green fluorescent protein (eGFP)-expression vectors were obtained: V1 (91% homology to HSPA6), V2 (five heat shock elements upstream the minimal HSPA6 promoter) and V3 (V1 and V2 combined). The highest eGFP transcription and translation levels were found in V3 transfected PC cells. The V3 promoter was used to control DTA, CRM176 and CRM197 expression, treatment response being investigated in four PC cell lines. DTAwt or CRM176 transfected cell growth was completely arrested after heat shock. CRM197 toxin presumed to be inactive, caused mild distress at 37 degrees C and induced a 25-50% reduction in cell growth after heat shock. Preliminary in vivo findings showed that heat treatment arrests tumor growth in DTA197 stably transfected PSN1 cells. In conclusion, the efficient HSP promoter identified in this study may be extremely useful in controlling the transcription of toxins such as CRM197, which have lethal dose-related effects, and may thus be a promising tool in PC gene therapy in vivo. PMID:19609296

  1. Microwave Heating Inactivates Shiga Toxin (Stx2) in Reconstituted Fat-Free Milk and Adversely Affects the Nutritional Value of Cell Culture Medium.

    PubMed

    Rasooly, Reuven; Hernlem, Bradley; He, Xiaohua; Friedman, Mendel

    2014-03-26

    Microwave exposure is a convenient and widely used method for defrosting, heating, and cooking numerous foods. Microwave cooking is also reported to kill pathogenic microorganisms that often contaminate food. In this study, we tested whether microwaves would inactivate the toxicity of Shiga toxin 2 (Stx2) added to 5% reconstituted fat-free milk administered to monkey kidney Vero cells. Heating of milk spiked with Stx2 in a microwave oven using a 10% duty cycle (cycle period of 30 s) for a total of 165 kJ energy or thermal heating (pasteurization), widely used to kill pathogenic bacteria, did not destroy the biological effect of the toxin in the Vero cells. However, conventional heating of milk to 95 C for 5 min or at an increased microwave energy of 198 kJ reduced the Stx2 activity. Gel electrophoresis showed that exposure of the protein toxin to high-energy microwaves resulted in the degradation of its original structure. In addition, two independent assays showed that exposure of the cell culture medium to microwave energy of 198 kJ completely destroyed the nutritional value of the culture medium used to grow the Vero cells, possibly by damaging susceptible essential nutrients present in the medium. These observations suggest that microwave heating has the potential to destroy the Shiga toxin in liquid food. PMID:24669932

  2. Randomized Clinical Trial Assessing the Safety and Immunogenicity of Oral Microencapsulated Enterotoxigenic Escherichia coli Surface Antigen 6 with or without Heat-Labile Enterotoxin with Mutation R192G▿

    PubMed Central

    Lapa, Joyce A.; Sincock, Stephanie A.; Ananthakrishnan, Madhumita; Porter, Chad K.; Cassels, Frederick J.; Brinkley, Carl; Hall, Eric R.; van Hamont, John; Gramling, Joseph D.; Carpenter, Colleen M.; Baqar, S.; Tribble, David R.

    2008-01-01

    An oral, microencapsulated anti-colonization factor 6 antigen (meCS6) vaccine, with or without heat-labile enterotoxin with mutation R192G (LTR192G) (mucosal adjuvant), against enterotoxigenic Escherichia coli (ETEC) was evaluated for regimen and adjuvant effects on safety and immunogenicity. Sixty subjects were enrolled into a three-dose, 2-week interval or four-dose, 2-day interval regimen. Each regimen was randomized into two equal groups of meCS6 alone (1 mg) or meCS6 with adjuvant (2 μg of LTR192G). The vaccine was well tolerated and no serious adverse events were reported. Serologic response to CS6 was low in all regimens (0 to 27%). CS6-immunogloublin A (IgA) antibody-secreting cell (ASC) responses ranged from 36 to 86%, with the highest level in the three-dose adjuvanted regimen; however, the magnitude was low. As expected, serologic and ASC LT responses were limited to adjuvanted regimens, with the exception of fecal IgA, which appeared to be nonspecific to LT administration. Further modifications to the delivery strategy and CS6 and adjuvant dose optimization will be needed before conducting further clinical trials with this epidemiologically important class of ETEC. PMID:18579693

  3. Evaluation of the immunogenicity of a transgenic tobacco plant expressing the recombinant fusion protein of GP5 of porcine reproductive and respiratory syndrome virus and B subunit of Escherichia coli heat-labile enterotoxin in pigs.

    PubMed

    Chia, Min-Yuan; Hsiao, Shih-Hsuan; Chan, Hui-Ting; Do, Yi-Yin; Huang, Pung-Ling; Chang, Hui-Wen; Tsai, Yi-Chieh; Lin, Chun-Ming; Pang, Victor Fei; Jeng, Chian-Ren

    2011-04-15

    Escherichia coli heat-labile enterotoxin B subunit (LTB) can be used as an adjuvant for co-administered antigens. Our previous study showed that the expression of neutralizing epitope GP5 of porcine reproductive and respiratory syndrome virus (PRRSV) in transgenic tobacco plant (GP5-T) could induce PRRSV-specific immune responses in pigs. A transgenic tobacco plant co-expressing LTB and PRRSV GP5 as a fusion protein (LTB-GP5-T) was further constructed and its immunogenicity was evaluated. Pigs were given orally three consecutive doses of equal concentration of recombinant GP5 protein expressed in leaves of LTB-GP5-T or GP5-T at a 2-week interval and challenged with PRRSV at 7 weeks post-initial immunization. Pigs receiving LTB-GP5-T or GP5-T developed PRRSV-specific antibody- and cell-mediated immunity and showed significantly lower viremia and tissue viral load and milder lung lesions than wild type tobacco plant (W-T). The LTB-GP5-T-treated group had relatively higher immune responses than the GP5-T-treated group, although the differences were not statistically significant. PMID:21277027

  4. Comparison of a live attenuated Salmonella Enteritidis vaccine candidate secreting Escherichia coli heat-labile enterotoxin B subunit with a commercial vaccine for efficacy of protection against internal egg contamination by Salmonella in hens

    PubMed Central

    Nandre, Rahul M.; Eo, Seong Kug; Park, Sang Youel; Lee, John Hwa

    2015-01-01

    This study compared a new live attenuated Salmonella Enteritidis vaccine candidate secreting Escherichia coli heat-labile enterotoxin B subunit (SE-LTB) with a commercial Salmonella Enteritidis (SE) vaccine for efficacy of protection against SE infection in laying hens. Chickens were divided into 3 groups of 20 each. Group A chickens were inoculated orally with phosphate-buffered saline and served as controls, group B chickens were inoculated orally with the vaccine candidate, and group C chickens were inoculated intramuscularly with a commercial vaccine, the primary inoculation in groups B and C being at 10 wk of age and the booster at 16 wk. Groups B and C showed significantly higher titers of plasma immunoglobulin G, intestinal secretory immunoglobulin A, and egg yolk immunoglobulin Y antibodies compared with the control group, and both vaccinated groups showed a significantly elevated cellular immune response. After virulent challenge, group B had significantly lower production of thin-shelled and/or malformed eggs and a significantly lower rate of SE contamination of eggs compared with the control group. Furthermore, the challenge strain was detected significantly less in all of the examined organs of group B compared with the control group. Group C had lower gross lesion scores only in the spleen and had lower bacterial counts only in the spleen, ceca, and ovary. These findings indicate that vaccination with the SE-LTB vaccine candidate can efficiently reduce internal egg and internal organ contamination by Salmonella and has advantages over the commercial vaccine. PMID:26130857

  5. Comparison of a live attenuated Salmonella Enteritidis vaccine candidate secreting Escherichia coli heat-labile enterotoxin B subunit with a commercial vaccine for efficacy of protection against internal egg contamination by Salmonella in hens.

    PubMed

    Nandre, Rahul M; Eo, Seong Kug; Park, Sang Youel; Lee, John Hwa

    2015-07-01

    This study compared a new live attenuated Salmonella Enteritidis vaccine candidate secreting Escherichia coli heat-labile enterotoxin B subunit (SE-LTB) with a commercial Salmonella Enteritidis (SE) vaccine for efficacy of protection against SE infection in laying hens. Chickens were divided into 3 groups of 20 each. Group A chickens were inoculated orally with phosphate-buffered saline and served as controls, group B chickens were inoculated orally with the vaccine candidate, and group C chickens were inoculated intramuscularly with a commercial vaccine, the primary inoculation in groups B and C being at 10 wk of age and the booster at 16 wk. Groups B and C showed significantly higher titers of plasma immunoglobulin G, intestinal secretory immunoglobulin A, and egg yolk immunoglobulin Y antibodies compared with the control group, and both vaccinated groups showed a significantly elevated cellular immune response. After virulent challenge, group B had significantly lower production of thin-shelled and/or malformed eggs and a significantly lower rate of SE contamination of eggs compared with the control group. Furthermore, the challenge strain was detected significantly less in all of the examined organs of group B compared with the control group. Group C had lower gross lesion scores only in the spleen and had lower bacterial counts only in the spleen, ceca, and ovary. These findings indicate that vaccination with the SE-LTB vaccine candidate can efficiently reduce internal egg and internal organ contamination by Salmonella and has advantages over the commercial vaccine. PMID:26130857

  6. Metal Complexes and Free Radical Toxins Produced by Pfiesteria piscicida

    SciTech Connect

    Moeller,P.; Beauchesne, K.; Huncik, K.; Davis, W.; Christopher, S.; Riggs-Gelasco, P.; Gelasco, A.

    2007-01-01

    Metal-containing organic toxins produced by Pfiesteria piscicida were characterized, for the first time, by corroborating data obtained from five distinct instrumental methods: nuclear magnetic resonance spectroscopy (NMR), inductively coupled plasma mass spectrometry (ICP-MS), liquid chromatography particle beam glow discharge mass spectrometry (LC/PB-GDMS), electron paramagnetic resonance spectroscopy (EPR), and X-ray absorption spectroscopy (XAS). The high toxicity of the metal-containing toxins is due to metal-mediated free radical production. This mode of activity explains the toxicity of Pfiesteria, as well as previously reported difficulty in observing the molecular target, due to the ephemeral nature of radical species. The toxins are highly labile in purified form, maintaining activity for only 2-5 days before all activity is lost. The multiple toxin congeners in active extracts are also susceptible to decomposition in the presence of white light, pH variations, and prolonged heat. These findings represent the first formal isolation and characterization of a radical forming toxic organic-ligated metal complex isolated from estuarine/marine dinoflagellates. These findings add to an increased understanding regarding the active role of metals interacting with biological systems in the estuarine environment, as well as their links and implications to human health.

  7. Metal Complexes And Free Radical Toxins Produced By Pfiesteria Piscicida

    SciTech Connect

    Moeller, P.D.R.; Beauchesne, K.R.; Huncik, K.M.; Davis, W.C.; Christopher, S.J.; Riggs-Gelasco, P.; Gelasco, A.K.

    2009-06-03

    Metal-containing organic toxins produced by Pfiesteria piscicida were characterized, for the first time, by corroborating data obtained from five distinct instrumental methods: nuclear magnetic resonance spectroscopy (NMR), inductively coupled plasma mass spectrometry (ICPMS), liquid chromatography particle beam glow discharge mass spectrometry (LC/PB-GDMS), electron paramagnetic resonance spectroscopy (EPR), and X-ray absorption spectroscopy (XAS). The high toxicity of the metal-containing toxins is due to metal-mediated free radical production. This mode of activity explains the toxicity of Pfiesteria, as well as previously reported difficulty in observing the molecular target, due to the ephemeral nature of radical species. The toxins are highly labile in purified form, maintaining activity for only 2-5 days before all activity is lost. The multiple toxin congeners in active extracts are also susceptible to decomposition in the presence of white light, pH variations, and prolonged heat. These findings represent the first formal isolation and characterization of a radical forming toxic organic-ligated metal complex isolated from estuarine/marine dinoflagellates. These findings add to an increased understanding regarding the active role of metals interacting with biological systems in the estuarine environment, as well as their links and implications to human health.

  8. Purification and characterization of salmolysin, an extracellular hemolytic toxin from Aeromonas salmonicida.

    PubMed Central

    Nomura, S; Fujino, M; Yamakawa, M; Kawahara, E

    1988-01-01

    An extracellular hemolytic toxin of Aeromonas salmonicida, termed salmolysin, was purified 945-fold by ammonium sulfate precipitation, anion-exchange chromatography on DEAE-cellulose, and gel filtration chromatography on Sephadex G-100 and Sepharose 2B. Salmolysin appeared homogeneous upon cellulose acetate membrane electrophoresis and immunodiffusion analysis. The molecular weight of the toxin was estimated to be approximately 200,000 by the sedimentation equilibrium method. The UV absorption spectrum showed a maximum at 275 nm and a minimum at 262 nm. The isoelectric point was found to be at pI 5.4. Carbohydrate and protein analyses and other biochemical data indicated that salmolysin is a glycoprotein, containing approximately 62% carbohydrates. The toxin is a heat-labile substance and is stable at a neutral pH value. Ferrous ion inhibited the activity, whereas metal-chelating agents did not affect the activity. Sulfhydryl reagents did not inhibit the toxin, whereas reducing agents, such as L-cysteine and reduced glutathione, inhibited the toxin to a certain extent. Salmolysin was inactivated by a nonionic detergent but was stimulated by an anionic detergent, sodium deoxycholate, at a low concentration. The toxin was also inactivated by subtilisin and trypsin but was not inhibited by papain and pepsin. Salmolysin, with a remarkable hemolytic activity against salmonid erythrocytes, was lethal to rainbow trout when it was injected intramuscularly. Images PMID:3136147

  9. Comparison of the mechanisms of action of cholera toxin and the heat-stable enterotoxins of Escherichia coli.

    PubMed Central

    Peterson, J W; Whipp, S C

    1995-01-01

    The mechanisms which enable cholera toxin (CT) and the Escherichia coli heat-stable enterotoxins (STa and STb) to stimulate intestinal secretion of water and electrolytes are only partially understood. CT evokes the synthesis of 3',5'-cyclic AMP (cAMP), and STa is known to elevate intestinal levels of 3',5'-cyclic GMP (cGMP). Neither of these recognized second messengers appears to mediate E. coli STb responses. We compared the secretory effects of CT, STa, and STb using the pig intestinal loop model and also measured the effects of toxin challenge on the synthesis of cAMP, cGMP, and prostaglandins (e.g., prostaglandin E2 [PGE2]), as well as on the release of 5-hydroxytryptamine (5-HT) from intestinal enterochromaffin cells. All three enterotoxins elicited fluid accumulation within a 2-h observation period. A combination of maximal doses of STa with STb yielded additive effects on fluid accumulation, which suggested different mechanisms of action for these toxins. Similarly, challenge of pig intestinal loops with a combination of CT and STb resulted in additive effects on fluid accumulation and luminal release of 5-HT. Unlike its effect on intestinal tissues from other animals, CT did not appear to elicit a dose-dependent cAMP response measurable in mucosal extracts from pig small intestine. In contrast, luminal fluid from CT-challenged pig intestinal loops contained dose-related amounts of cAMP and PGE2 that had been secreted from the mucosa. cAMP responses to STa or STb could not be demonstrated in either mucosal tissue or luminal fluid. In contrast, cGMP levels were increased in the intestinal fluid of loops challenged with STa but not in those challenged with STb. While the mechanisms of action of CT and STa are thought to involve impulse transmission via the enteric nervous system, we demonstrated significant stimulation of PGE2 synthesis and 5-HT release for CT and STb but very little for STa. We conclude from these data that the mechanisms of action of STa, STb, and CT are distinct, although the mode of action of STb may have some similarity to that of CT. Since STb stimulated the release of both PGE2 and 5-HT from the intestinal mucosa, the data suggested the potential for an effect of STb on the enteric nervous system. PMID:7890409

  10. The Divergent CD8+ T Cell Adjuvant Properties of LT-IIb and LT-IIc, Two Type II Heat-Labile Enterotoxins, Are Conferred by Their Ganglioside-Binding B Subunits

    PubMed Central

    Hu, John C.; Greene, Christopher J.; King-Lyons, Natalie D.; Connell, Terry D.

    2015-01-01

    Poor immune responses elicited by vaccine antigens can be enhanced by the use of appropriate adjuvants. Type II heat-labile enterotoxins (HLT) produced by Escherichia coli are extremely potent adjuvants that augment both humoral and cellular immunity to co-administered antigens. Recent findings demonstrate that LT-IIb and LT-IIc, two type II HLT adjuvants, exhibit potent, yet distinguishable CD8+ T cell adjuvant properties. While LT-IIc elicits a robust and rapid response at one week after administration, LT-IIb engenders a more gradual and slower expansion of antigen-specific CD8+ T cells that correlates with improved immunity. The variations in immune effects elicited by the HLT adjuvants have been generally attributed to their highly divergent B subunits that mediate binding to various gangliosides on cell surfaces. Yet, HLT adjuvants with point mutations in the B subunit that significantly alter ganglioside binding retain similar adjuvant functions. Therefore, the contribution of the B subunits to adjuvanticity remains unclear. To investigate the influence of the B subunits on the enhancement of immune responses by LT-IIb and LT-IIc, chimeric HLT were engineered in which the B subunits of the two adjuvants were exchanged. Comparing the immune potentiating characteristics of both native and chimeric HLT adjuvants, it was found that not all the adjuvant characteristics of the HLT adjuvants were modulated by the respective B subunits. Specifically, the differences in the CD8+ T cell kinetics and protective responses elicited by LT-IIb and LT-IIc did indeed followed their respective B subunits. However, induction of IL-1 from macrophages and the capacity to intoxicate cells in a mouse Y1 adrenal cell bioassay did not correlate with the B subunits. Therefore, it is likely that additional factors other than the B subunits contribute to the effects elicited by the HLT adjuvants. PMID:26565800

  11. Intranasal immunization against herpes simplex virus infection by using a recombinant glycoprotein D fused with immunomodulating proteins, the B subunit of Escherichia coli heat-labile enterotoxin and interleukin-2.

    PubMed Central

    Hazama, M; Mayumi-Aono, A; Miyazaki, T; Hinuma, S; Fujisawa, Y

    1993-01-01

    To establish a novel strategy of mucosal immunization against herpes simplex virus type 1 (HSV-1) infection, we studied the immune responses elicited by intranasal immunization with several forms of a recombinant glycoprotein D (gD) of HSV-1. A truncated gD (t-gD) co-administered with heat-labile enterotoxin B subunit (LTB) from Escherichia coli induced both a mucosal immune response involving secretion of anti-gD IgA and serum IgG production. The levels of these responses are comparable to those in mice which have recovered from intranasal HSV-1 infections. The fusion protein (t-gD-LTB), consisting of t-gD and LTB, induced the responses more efficiently than did co-administration of t-gD and LTB, although GM1 ganglioside binding activity was significantly reduced in t-gD-LTB. We found that another fusion protein, consisting of t-gD and human interleukin-2 (t-gD-IL-2), also elicited antibody responses comparable to those induced by t-gD-LTB. Immunity acquired by intranasal immunization with t-gD-IL-2 protected mice from intraperitoneal HSV-1 infections, whereas t-gD-LTB or t-gD alone failed to provide protection against infection. Even in a mouse strain that responded highly to subcutaneously administered gD, intranasally administered t-gD did not elicit antibody responses. The lack of response to gD was clearly abrogated by co-administration with IL-2, and administration of t-gD-IL-2 induced an excellent level of antibody responses in this strain. These results suggest that the IL-2 fusion strategy yields a new type of mucosal immunization, the mechanism of which differs from that speculated for the mucosal adjuvant activity of LTB. Images Figure 1 PMID:8388365

  12. Labile neurotoxin in serum of calves with "nervous" coccidiosis.

    PubMed Central

    Isler, C M; Bellamy, J E; Wobeser, G A

    1987-01-01

    Mouse inoculation was used to test for the presence of a toxin in the serum, cerebrospinal fluid, and intestinal contents collected from cases of bovine enteric coccidiosis, with and without neurological signs, and from control calves. Intravenous inoculation of mice with 10 mL/kg of serum from calves showing nervous signs caused effects significantly different from those caused by the inoculation of serum from calves not showing nervous signs and from control calves. The effect was particularly evident in female mice. At this dosage severe neurological signs such as loss of righting reflex, seizures and death occurred only with serum from calves with "nervous coccidiosis". The results suggest that serum from the calves with neurological signs contains a neurotoxin. This toxin appears to be highly labile. It was not present in the cerebrospinal fluid at levels comparable to those in the serum. The significance of this labile neurotoxin with respect to the pathogenesis of the neurological signs associated with bovine enteric coccidiosis is unknown. PMID:2955865

  13. Cholera toxin B subunits assemble into pentamers--proposition of a fly-casting mechanism.

    PubMed

    Zrimi, Jihad; Ng Ling, Alicia; Giri-Rachman Arifin, Ernawati; Feverati, Giovanni; Lesieur, Claire

    2010-01-01

    The cholera toxin B pentamer (CtxB(5)), which belongs to the AB(5) toxin family, is used as a model study for protein assembly. The effect of the pH on the reassembly of the toxin was investigated using immunochemical, electrophoretic and spectroscopic methods. Three pH-dependent steps were identified during the toxin reassembly: (i) acquisition of a fully assembly-competent fold by the CtxB monomer, (ii) association of CtxB monomer into oligomers, (iii) acquisition of the native fold by the CtxB pentamer. The results show that CtxB(5) and the related heat labile enterotoxin LTB(5) have distinct mechanisms of assembly despite sharing high sequence identity (84%) and almost identical atomic structures. The difference can be pinpointed to four histidines which are spread along the protein sequence and may act together. Thus, most of the toxin B amino acids appear negligible for the assembly, raising the possibility that assembly is driven by a small network of amino acids instead of involving all of them. PMID:21203571

  14. Intradermal administration of the Type II heat-labile enterotoxins LT-IIb and LT-IIc of enterotoxigenic Escherichia coli enhances humoral and CD8+ T cell immunity to a co-administered antigen.

    PubMed

    Hu, John C; Mathias-Santos, Camila; Greene, Christopher J; King-Lyons, Natalie D; Rodrigues, Juliana F; Hajishengallis, George; Ferreira, Lus C S; Connell, Terry D

    2014-01-01

    Vaccinations are extremely effective at combating infectious diseases. Many conserved antigen (Ag) targets, however, are poorly immunogenic. Protein subunit vaccines frequently elicit only humoral immune responses and fail to confer protection against serious intracellular pathogens. These barriers to vaccine development are often overcome by the use of appropriate adjuvants. Heat-labile enterotoxins (HLT) produced by enterotoxigenic strains of Escherichia coli are potent adjuvants when administered by mucosal or systemic routes. The efficacy of the type II HLT, however, has not been well-defined when administered by the intradermal (ID) route. Using a murine ID immunization model, the adjuvant properties of LT-IIb and LT-IIc, two type II HLTs, were compared with those of LT-I, a prototypical type I HLT. While all three HLT adjuvants enhanced Ag-specific humoral responses to similar levels, LT-IIb and LT-IIc, in contrast to LT-I, induced a more vigorous Ag-specific CD8+ T cell response and proffered faster clearance of Listeria monocytogenes in a challenge model. Additionally, LT-IIb and LT-IIc induced distinct differences in the profiles of the Ag-specific CD8+ T cell responses. While LT-IIc stimulated a robust and rapid primary CD8+ T cell response, LT-IIb exhibited slower CD8+ T cell expansion and contraction kinetics with the formation of higher percentages of effector memory cells. In comparison to LT-I and LT-IIc, LT-IIb evoked better long-term protection after immunization. Furthermore, LT-IIb and LT-IIc enhanced the total number of dendritic cells (DC) in the draining lymph node (DLN) and expression of costimulatory molecules CD80, CD86, and CD40 on DCs. In contrast to LT-I, LT-IIb and LT-IIc induced less edema, cellular infiltrates, and general inflammation at the site of ID injection. Thus, LT-IIb and LT-IIc are attractive comprehensive ID adjuvants with unique characteristic that enhance humoral and cellular immunity to a co-administered protein Ag. PMID:25536061

  15. Structureactivity correlations of variant forms of the B pentamer of Escherichia coli type II heat-labile enterotoxin LT-IIb with Toll-like receptor 2 binding

    PubMed Central

    Cody, Vivian; Pace, Jim; Nawar, Hesham F.; King-Lyons, Natalie; Liang, Shuang; Connell, Terry D.; Hajishengallis, George

    2012-01-01

    The pentameric B subunit of the type II heat-labile enterotoxin of Escherichia coli (LT-IIb-B5) is a potent signaling molecule capable of modulating innate immune responses. It has previously been shown that LT-IIb-B5, but not the LT-IIb-B5 Ser74Asp variant [LT-IIb-B5(S74D)], activates Toll-like receptor (TLR2) signaling in macrophages. Consistent with this, the LT-IIb-B5(S74D) variant failed to bind TLR2, in contrast to LT-IIb-B5 and the LT-IIb-B5 Thr13Ile [LT-IIb-B5(T13I)] and LT-IIb-B5 Ser74Ala [LT-IIb-B5(S74A)] variants, which displayed the highest binding activity to TLR2. Crystal structures of the Ser74Asp, Ser74Ala and Thr13Ile variants of LT-IIb-B5 have been determined to 1.90, 1.40 and 1.90? resolution, respectively. The structural data for the Ser74Asp variant reveal that the carboxylate side chain points into the pore, thereby reducing the pore size compared with that of the wild-type or the Ser74Ala variant B pentamer. On the basis of these crystallographic data, the reduced TLR2-binding affinity of the LT-IIb-B5(S74D) variant may be the result of the pore of the pentamer being closed. On the other hand, the explanation for the enhanced TLR2-binding activity of the LT-IIb-B5(S74A) variant is more complex as its activity is greater than that of the wild-type B pentamer, which also has an open pore as the Ser74 side chain points away from the pore opening. Data for the LT-IIb-B5(T13I) variant show that four of the five variant side chains point to the outside surface of the pentamer and one residue points inside. These data are consistent with the lack of binding of the LT-IIb-B5(T13I) variant to GD1a ganglioside. PMID:23151625

  16. LT-IIc, a new member of the type II heat-labile enterotoxin family, exhibits potent immunomodulatory properties that are different from those induced by LT-IIa or LT-IIb

    PubMed Central

    Nawar, Hesham F.; Greene, Christopher J.; Lee, Chang Hoon; Mandell, Lorrie; Connell, Terry D.

    2010-01-01

    A plethora of human pathogens invade and/or colonize mucosal surfaces. Elaboration of strong, protective immune responses against those pathogens by mucosal vaccination, however, is hampered by endogenous regulatory systems in the mucosae that dampen responses to foreign antigens (Ag). To overcome those natural barriers, mucosal adjuvants must be employed. Using a mouse mucosal immunization model and AgI/II, a weak immunogen from Streptococcus mutans, LT-IIc, a new member of the type II subgroup of the heat-labile enterotoxin family, was shown to have potent mucosal adjuvant properties. In comparison to mice intranasally immunized only with AgI/II, co-administration of AgI/II with LT-IIc enhanced production of Ag-specific IgA antibodies in the saliva and vaginal fluids and Ag-specific IgA and IgG in the serum. Secretion of IL-2, IL-6, IL-17, IFN-γ, and TNF-α was enhanced in cultures of AgI/II-stimulated splenic cells isolated from mice that had received LT-IIc as a mucosal adjuvant. In contrast, secretion of IL-10 was suppressed in those cells. This pattern of cytokine secretion suggested that LT-IIc stimulates both Th1 and Th2 immune responses. In contrast to LT-IIa and LT-IIb, the original members of the type II subgroup that also are mucosal adjuvants, LT-IIc dramatically enhanced secretion of IL-1α and IL-1β in peritoneal macrophages that had been co-cultured with LPS. Furthermore, the B pentameric subunit of LT-IIc augmented uptake of Ag by bone marrow-derived dendritic cells to levels that exceeded those attained by use of LPS or by the B pentamers of LT-IIa or LT-IIb. These data confirmed that LT-IIc is a strong mucosal adjuvant with immunomodulatory properties that are distinguishable from those of LT-IIa and LT-IIb and which has immunomodulatory properties that may be exploitable in vaccine development. PMID:21095251

  17. Electrodermal lability in anxiety disorders.

    PubMed

    Jensen, H H; Hasle, N; Birket-Smith, M

    1996-03-01

    Twenty-eight anxiety patients, aged below 50 years, were diagnosed according to DSM-III-R criteria (panic disorder with and without agoraphobia, generalised anxiety disorder, and anxiety disorder not otherwise specified). The patients were characterised by high levels of state and trait anxiety and neuroticism, compared with the controls. However, there were no differences between patients and controls in electrodermal habituation rate, non-specific activity, or skin resistance level. When the patients were divided into electrodermally labile and stable subjects, significant differences were found between patients and controls in both electrodermal activity and Eysenck's personality dimensions. The labile patients were more introverted and attained higher psychoticism scores than either the stable patients or controls. Duration of anxiety symptoms removed the difference found in extroversion, but not in any other variable. The results are discussed in relation to the utility of electrodermal measurements in validation of diagnostic entities. It is concluded, that from the psychophysiological point of view, anxiety disorders may be examined within a dimensional framework. PMID:8900823

  18. Heat treatment and the use of additives to improve the stability of paralytic shellfish poisoning toxins in shellfish tissue reference materials for internal quality control and proficiency testing.

    PubMed

    Burrell, Stephen; Clion, Valentin; Auroy, Virginie; Foley, Barry; Turner, Andrew D

    2015-06-01

    The need for homogenous reference materials stable for paralytic shellfish toxins is vital for the monitoring and quality assurance of these potent neurotoxins in shellfish. Two stabilisation techniques were investigated, heat treatment through autoclaving and the addition of preserving additives into the tissue matrix. Short and long-term stability experiments as well as homogeneity determination were conducted on materials prepared by both techniques in comparison with an untreated control using two LC-FLD methods. Both techniques improved the stability of the matrix and the PSP toxins present compared to the controls. A material was prepared using the combined techniques of heat treatment followed by spiking with additives and data is presented from this optimised reference material as used over a two year period in the Irish national monitoring program and in a development exercise as part of a proficiency testing scheme operated by QUASIMEME (Quality Assurance of Information for Marine Environmental Monitoring in Europe) since 2011. The results were indicative of the long-term stability of the material as evidenced through consistent assigned values in the case of the proficiency testing scheme and a low relative standard deviation of 10.5% for total toxicity data generated over 24 months. PMID:25816999

  19. Effect of water activity and pH on growth and toxin production by Clostridium botulinum type G.

    PubMed Central

    Briozzo, J; de Lagarde, E A; Chirife, J; Parada, J L

    1986-01-01

    The combined effect of water activity (aw) and pH on growth and toxin production by Clostridium botulinum type G strain 89 was investigated. The minimum aw at which growth and toxin formation occurred was 0.965, for media in which the pH was adjusted with either sodium chloride or sucrose. The minimum pH (at the optimum aw) for growth and toxin production of C. botulinum type G was found to be 5.6. Optimum conditions for toxin activation were a trypsin concentration of 0.1%, a pH of the medium of 6.5, and an incubation for 45 min at 37 degrees C. These data did not show evidence of heat-labile spores, since a heat shock of 75 degrees C for 10 min did not significantly decrease the spore count of strain 89G in media at pH 7.0 or 5.6. It was frequently observed that cells grown at reduced aw or pH experienced severe morphological changes. PMID:3518631

  20. A toxin-antitoxin system encoded by the Xylella fastidiosa chromosome regulates growth

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacteria encode toxin-antitoxin (TA) systems consisting of a stable toxin and a cognate labile antitoxin. When encoded by a plasmid, TA systems confer stable plasmid inheritance. When encoded by the chromosome, TA systems may confer advantageous responses to environmental stress. The chromosome of...

  1. [The effect of repeated amputations on planarian regeneration in the presence of the heat-stable toxin from Bacillus thuringiensis].

    PubMed

    Ziller, C

    1976-09-13

    The exotoxin of Bacillus thuringiensis, which is an inhibitor of RNA synthesis, inhibits Planarian regeneration. Planarians which have been cut twice at the same level are able to regenerate after the second section in the presence of the toxin. This indicates that the first amputation stimulates a synthesis of stable RNAs which thus are available for regeneration at the moment of the second section. PMID:825282

  2. Pertussis toxin

    SciTech Connect

    Sekura, R.D.; Moss, J.; Vaughan, M.

    1985-01-01

    This book contains 13 selections. Some of the titles are: Genetic and Functional Studies of Pertussis Toxin Substrates; Effect of Pertussis Toxin on the Hormonal Responsiveness of Different Tissues; Extracellular Adenylate Cyclase of Bordetella pertussis; and GTP-Regulatory Proteins are Introcellular Messagers: A Model for Hormone Action.

  3. Millipede toxin

    MedlinePLUS

    ... plenty of soap and water. Do NOT use alcohol to wash the area. Wash eyes with plenty of water if any toxin gets in them. Get medical attention right away. Tell the health care provider if any toxin got in the eyes.

  4. Mucosal adjuvants and anti-infection and anti-immunopathology vaccines based on cholera toxin, cholera toxin B subunit and CpG DNA.

    PubMed

    Holmgren, Jan; Harandi, Ali M; Czerkinsky, Cecil

    2003-04-01

    The mucosal immune system consists of an integrated network of lymphoid cells that work in concert with innate host factors to promote host defence. Mucosal immunization can be used both to protect the mucosal surfaces against colonization and invasion by microbial pathogens and to provide a means for immunological treatment of selected autoimmune, allergic or infectious-immunopathological disorders through the induction of antigen-specific tolerance. The development of mucosal vaccines, whether for prevention of infectious diseases or for oral tolerance immunotherapy, requires efficient antigen delivery and adjuvant systems. Significant progress has recently been made to generate partly or wholly detoxified derivatives of cholera toxin (including the completely nontoxic cholera toxin B subunit) and the closely related Escherichia coli heat-labile enterotoxin, with retained adjuvant activity. Cholera toxin B subunit is a protective component of a widely registered oral vaccine against cholera, and has proven to be a promising vector for either giving rise to anti-infective immunity or for inducing peripheral anti-inflammatory tolerance to chemically or genetically linked foreign antigens administered mucosally. Promising advances have also recently been made in the design of efficient mucosal adjuvants based on bacterial DNA that contains CpG-motifs and various imidazoquinoline compounds binding to different Toll-like receptors on mucosal antigen-presenting cells. PMID:12899572

  5. Labile sulfide and sulfite in phytochelatin complexes

    SciTech Connect

    Eannetta, N.T.; Steffens, J.C. )

    1989-04-01

    Heavy metals such as cadmium induce tomato cell cultures to synthesize the metal binding polypeptides ({gamma}-Glu-Cys){sub 3} and ({gamma}-Glu-Cys){sub 4}-Gly (phytochelatins). Tomato cells selected for growth on normally lethal concentrations of CdCl{sub 2} synthesize higher quantities of these polypeptides. Cd{sup r} cells are not cross-resistant to other heavy metals, and recent work suggests that metal detoxification by these peptides may be Cd-specific. The occurrence of labile sulfur as a component of the metal complex raises questions concerning possible functions of phytochelatins besides that of Cd binding. The presence of acid-labile sulfide ion in phytochelatin complexes has been reported by several groups. We report the additional finding that labile sulfite is also present in these complexes and in higher amounts than sulfide. Sulfide and sulfite are both released from the metal binding complex by acidification or by treatment with EDTA.

  6. BOTULINUM TOXIN

    PubMed Central

    Nigam, P K; Nigam, Anjana

    2010-01-01

    Botulinum toxin, one of the most poisonous biological substances known, is a neurotoxin produced by the bacterium Clostridium botulinum. C. botulinum elaborates eight antigenically distinguishable exotoxins (A, B, C1, C2, D, E, F and G). All serotypes interfere with neural transmission by blocking the release of acetylcholine, the principal neurotransmitter at the neuromuscular junction, causing muscle paralysis. The weakness induced by injection with botulinum toxin A usually lasts about three months. Botulinum toxins now play a very significant role in the management of a wide variety of medical conditions, especially strabismus and focal dystonias, hemifacial spasm, and various spastic movement disorders, headaches, hypersalivation, hyperhidrosis, and some chronic conditions that respond only partially to medical treatment. The list of possible new indications is rapidly expanding. The cosmetological applications include correction of lines, creases and wrinkling all over the face, chin, neck, and chest to dermatological applications such as hyperhidrosis. Injections with botulinum toxin are generally well tolerated and side effects are few. A precise knowledge and understanding of the functional anatomy of the mimetic muscles is absolutely necessary to correctly use botulinum toxins in clinical practice. PMID:20418969

  7. Botulinum Toxin Therapy

    MedlinePLUS

    ... Diseases and treatments A - D Botulinum toxin Botulinum toxin therapy Also called botulinum rejuvenation Brand names: Botox ... what your policy covers. Learn more about botulinum toxin therapy: Is botulinum toxin therapy the right choice ...

  8. Lability of Humic-Bound Phosphorus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phosphorus (P) has long been known to be present in humic substances from various sources. However, information on the lability of humic-bound P is very limited although such information is critical for understanding the role of humic substances in P cycling and nutrition. In this presentation, we d...

  9. [Marine toxins].

    PubMed

    Lueger, A; Scherr, D; Lang, B; Brodmann, M; Stark, G

    1999-01-01

    The consumption of seafood, which is contaminated by toxines of red tides, is a common cause of disease in tropic regions. The most important diseases, which are caused by red tides are Paralytic Shellfish Poisoning (PSP), Diarrhetic Shellfish Poisoning (DSP), Neurotoxic Shellfish Poisoning (NSP), Amnesic Shellfish Poisoning (ASP), Ciguatera Fish Poisoning (CFP). PMID:11315409

  10. Red alert: labile heme is an alarmin.

    PubMed

    Soares, Miguel P; Bozza, Marcelo T

    2016-02-01

    Alarmins are a heterogeneous group of endogenous molecules that signal cellular damage when sensed extracellularly. Heme is an endogenous molecule that acts as a prosthetic group of hemoproteins, such as hemoglobin and myoglobin. When released from damaged red blood cells or muscle cells, oxidized hemoglobin and myoglobin release their prosthetic heme groups, respectively. This generates labile heme, which is sensed by pattern recognition receptors (PRR) expressed by innate immune cells and possibly regulatory T cells (TREG). The ensuing adaptive response, which alerts for the occurrence of red blood cell or muscle cell damage, regulates the pathologic outcome of hemolysis or rhabdomyolysis, respectively. In conclusion, we propose that labile heme is an alarmin. PMID:26741528

  11. Transcutaneous immunization with cross-reacting material CRM(197) of diphtheria toxin boosts functional antibody levels in mice primed parenterally with adsorbed diphtheria toxoid vaccine.

    PubMed

    Stickings, Paul; Peyre, Marisa; Coombes, Laura; Muller, Sylviane; Rappuoli, Rino; Del Giudice, Giuseppe; Partidos, Charalambos D; Sesardic, Dorothea

    2008-04-01

    Transcutaneous immunization (TCI) capitalizes on the accessibility and immunocompetence of the skin, elicits protective immunity, simplifies vaccine delivery, and may be particularly advantageous when frequent boosting is required. In this study we examined the potential of TCI to boost preexisting immune responses to diphtheria in mice. The cross-reacting material (CRM(197)) of diphtheria toxin was used as the boosting antigen and was administered alone or together with either one of two commonly used mucosal adjuvants, cholera toxin (CT) and a partially detoxified mutant of heat-labile enterotoxin of Escherichia coli (LTR72). We report that TCI with CRM(197) significantly boosted preexisting immune responses elicited after parenteral priming with aluminum hydroxide-adsorbed diphtheria toxoid (DTxd) vaccine. In the presence of LTR72 as an adjuvant, toxin-neutralizing antibody titers were significantly higher than those elicited by CRM(197) alone and were comparable to the functional antibody levels induced after parenteral booster immunization with the adsorbed DTxd vaccine. Time course study showed that high levels of toxin-neutralizing antibodies persisted for at least 14 weeks after the transcutaneous boost. In addition, TCI resulted in a vigorous antigen-specific proliferative response in all groups of mice boosted with the CRM(197) protein. These findings highlight the promising prospect of using booster administrations of CRM(197) via the transcutaneous route to establish good herd immunity against diphtheria. PMID:18227167

  12. Transcutaneous Immunization with Cross-Reacting Material CRM197 of Diphtheria Toxin Boosts Functional Antibody Levels in Mice Primed Parenterally with Adsorbed Diphtheria Toxoid Vaccine▿

    PubMed Central

    Stickings, Paul; Peyre, Marisa; Coombes, Laura; Muller, Sylviane; Rappuoli, Rino; Del Giudice, Giuseppe; Partidos, Charalambos D.; Sesardic, Dorothea

    2008-01-01

    Transcutaneous immunization (TCI) capitalizes on the accessibility and immunocompetence of the skin, elicits protective immunity, simplifies vaccine delivery, and may be particularly advantageous when frequent boosting is required. In this study we examined the potential of TCI to boost preexisting immune responses to diphtheria in mice. The cross-reacting material (CRM197) of diphtheria toxin was used as the boosting antigen and was administered alone or together with either one of two commonly used mucosal adjuvants, cholera toxin (CT) and a partially detoxified mutant of heat-labile enterotoxin of Escherichia coli (LTR72). We report that TCI with CRM197 significantly boosted preexisting immune responses elicited after parenteral priming with aluminum hydroxide-adsorbed diphtheria toxoid (DTxd) vaccine. In the presence of LTR72 as an adjuvant, toxin-neutralizing antibody titers were significantly higher than those elicited by CRM197 alone and were comparable to the functional antibody levels induced after parenteral booster immunization with the adsorbed DTxd vaccine. Time course study showed that high levels of toxin-neutralizing antibodies persisted for at least 14 weeks after the transcutaneous boost. In addition, TCI resulted in a vigorous antigen-specific proliferative response in all groups of mice boosted with the CRM197 protein. These findings highlight the promising prospect of using booster administrations of CRM197 via the transcutaneous route to establish good herd immunity against diphtheria. PMID:18227167

  13. Biologically labile photoproducts from riverine non-labile dissolved organic carbon in the coastal waters

    NASA Astrophysics Data System (ADS)

    Kasurinen, V.; Aarnos, H.; Vähätalo, A.

    2015-06-01

    In order to assess the production of biologically labile photoproducts (BLPs) from non-labile riverine dissolved organic carbon (DOC), we collected water samples from ten major rivers, removed labile DOC and mixed the residual non-labile DOC with artificial seawater for microbial and photochemical experiments. Bacteria grew on non-labile DOC with a growth efficiency of 11.5% (mean; range from 3.6 to 15.3%). Simulated solar radiation transformed a part of non-labile DOC into BLPs, which stimulated bacterial respiration and production, but did not change bacterial growth efficiency (BGE) compared to the non-irradiated dark controls. In the irradiated water samples, the amount of BLPs stimulating bacterial production depended on the photochemical bleaching of chromophoric dissolved organic matter (CDOM). The apparent quantum yields for BLPs supporting bacterial production ranged from 9.5 to 76 (mean 39) (μmol C mol photons-1) at 330 nm. The corresponding values for BLPs supporting bacterial respiration ranged from 57 to 1204 (mean 320) (μmol C mol photons-1). According to the calculations based on spectral apparent quantum yields and local solar radiation, the annual production of BLPs ranged from 21 (St. Lawrence) to 584 (Yangtze) mmol C m-2 yr-1 in the plumes of the examined rivers. Complete photobleaching of riverine CDOM in the coastal ocean was estimated to produce 10.7 Mt C BLPs yr-1 from the rivers examined in this study and globally 38 Mt yr-1 (15% of riverine DOC flux from all rivers), which support 4.1 Mt yr-1 of bacterial production and 33.9 Mt yr-1 bacterial respiration.

  14. Transposition of the Heat-Stable Toxin astA Gene into a Gifsy-2-Related Prophage of Salmonella enterica Serovar Abortusovis

    PubMed Central

    Bacciu, Donatella; Falchi, Giovanni; Spazziani, Alessandra; Bossi, Lionello; Marogna, Gavino; Leori, Guido Sisinnio; Rubino, Salvatore; Uzzau, Sergio

    2004-01-01

    The horizontal transfer and acquisition of virulence genes via mobile genetic elements have been a major driving force in the evolution of Salmonella pathogenicity. Serovars of Salmonella enterica carry variable assortments of phage-encoded virulence genes, suggesting that temperate phages play a pivotal role in this process. Epidemic isolates of S. enterica serovar Typhimurium are consistently lysogenic for two lambdoid phages, Gifsy-1 and Gifsy-2, carrying known virulence genes. Other serovars of S. enterica, including serovars Dublin, Gallinarum, Enteritidis, and Hadar, carry distinct prophages with similarity to the Gifsy phages. In this study, we analyzed Gifsy-related loci from S. enterica serovar Abortusovis, a pathogen associated exclusively with ovine infection. A cryptic prophage, closely related to serovar Typhimurium phage Gifsy-2, was identified. This element, named Gifsy-2AO, was shown to contribute to serovar Abortusovis systemic infection in lambs. Sequence analysis of the prophage b region showed a large deletion which covers genes encoding phage tail fiber proteins and putative virulence factors, including type III secreted effector protein SseI (GtgB, SrfH). This deletion was identified in most of the serovar Abortusovis isolates tested and might be dependent on the replicative transposition of an adjacent insertion sequence, IS1414, previously identified in pathogenic Escherichia coli strains. IS1414 encodes heat-stable toxin EAST1 (astA) and showed multiple genomic copies in isolates of serovar Abortusovis. To our knowledge, this is the first evidence of intergeneric transfer of virulence genes via insertion sequence elements in Salmonella. The acquisition of IS1414 (EAST1) and its frequent transposition within the chromosome might improve the fitness of serovar Abortusovis within its narrow ecological niche. PMID:15231789

  15. Transposition of the heat-stable toxin astA gene into a gifsy-2-related prophage of Salmonella enterica serovar Abortusovis.

    PubMed

    Bacciu, Donatella; Falchi, Giovanni; Spazziani, Alessandra; Bossi, Lionello; Marogna, Gavino; Leori, Guido Sisinnio; Rubino, Salvatore; Uzzau, Sergio

    2004-07-01

    The horizontal transfer and acquisition of virulence genes via mobile genetic elements have been a major driving force in the evolution of Salmonella pathogenicity. Serovars of Salmonella enterica carry variable assortments of phage-encoded virulence genes, suggesting that temperate phages play a pivotal role in this process. Epidemic isolates of S. enterica serovar Typhimurium are consistently lysogenic for two lambdoid phages, Gifsy-1 and Gifsy-2, carrying known virulence genes. Other serovars of S. enterica, including serovars Dublin, Gallinarum, Enteritidis, and Hadar, carry distinct prophages with similarity to the Gifsy phages. In this study, we analyzed Gifsy-related loci from S. enterica serovar Abortusovis, a pathogen associated exclusively with ovine infection. A cryptic prophage, closely related to serovar Typhimurium phage Gifsy-2, was identified. This element, named Gifsy-2AO, was shown to contribute to serovar Abortusovis systemic infection in lambs. Sequence analysis of the prophage b region showed a large deletion which covers genes encoding phage tail fiber proteins and putative virulence factors, including type III secreted effector protein SseI (GtgB, SrfH). This deletion was identified in most of the serovar Abortusovis isolates tested and might be dependent on the replicative transposition of an adjacent insertion sequence, IS1414, previously identified in pathogenic Escherichia coli strains. IS1414 encodes heat-stable toxin EAST1 (astA) and showed multiple genomic copies in isolates of serovar Abortusovis. To our knowledge, this is the first evidence of intergeneric transfer of virulence genes via insertion sequence elements in Salmonella. The acquisition of IS1414 (EAST1) and its frequent transposition within the chromosome might improve the fitness of serovar Abortusovis within its narrow ecological niche. PMID:15231789

  16. Mucosal Immunization with Helicobacter, CpG DNA, and Cholera Toxin Is Protective

    PubMed Central

    Jiang, Weiwen; Baker, Henry J.; Smith, Bruce F.

    2003-01-01

    The mucosal delivery of antigens requires an effective adjuvant to induce mucosal immunity. Current mucosal adjuvants include cholera toxin (CT) and Escherichia coli heat-labile toxin. Unmethylated CpG immunostimulatory oligodeoxynucleotides (ODNs) have been proposed as novel mucosal adjuvants. In this study, mice were immunized with sonicated Helicobacter felis with CT and/or CpG ODN adjuvants. All groups receiving either adjuvant singly or in combination developed increased serum anti-H. felis immunoglobulin G (IgG). The addition of either CpG or CT, or both, produced a specific fecal anti-H. felis IgA response, with the highest IgA levels occurring in animals immunized intranasally with sonicated H. felis with CT and CpG. Following H. felis challenge, addition of the adjuvant CpG ODN provided no significant protection, while groups given CT showed a high degree of protection, although not complete. When CpG ODN was combined with CT and the vaccine combination was delivered intranasally, no bacterial colonization was detected by quantitative PCR, providing sterile immunity and demonstrating synergy between CpG ODN and CT. PMID:12496147

  17. The 2.3 {angstrom} crystal structure of cholera toxin B subunit pentamer: Choleragenoid

    SciTech Connect

    Zhang, Rong-Guang; Westbrook, M.L.; Maulik, P.R.; Reed, R.A.; Shipley, G.; Westbrook, E.M.; Scott, D.L.; Otwinowski, Z.

    1996-02-01

    Cholera toxin, a heterohexameric AB{sub 5} enterotoxin released by Vibrio cholera, induces a profuse secretory diarrhea in susceptible hosts. Choleragenoid, the B subunit pentamer of cholera toxin, directs the enzymatic A subunit to its target by binding to GM{sub 1} gangliosides exposed on the luminal surface of intestinal epithelial cells. We have solved the crystal structure of choleragenoid at 2.3 {Angstrom} resolution by combining single isomorphous replacement with non-crystallographic symmetry averaging. The structure of the B subunits, and their pentameric arrangement, closely resembles that reported for the intact holotoxin (choleragen), the heat-labile enterotoxin from E. coli, and for a choleragenoid-GM{sub 1} pentasaccharide complex. In the absence of the A subunit the central cavity of the B pentamer is a highly solvated channel. The binding of the A subunit or the receptor pentasaccharide to choleragenoid has only a modest effect on the local stereochemistry and does not perceptibly alter the subunit interface.

  18. Mucosal adjuvants and anti-infection and anti-immunopathology vaccines based on cholera toxin, cholera toxin B subunit and CpG DNA.

    PubMed

    Holmgren, Jan; Adamsson, Jenni; Anjuère, Fabienne; Clemens, John; Czerkinsky, Cecil; Eriksson, Kristina; Flach, Carl-Fredrik; George-Chandy, Annie; Harandi, Ali M; Lebens, Michael; Lehner, Thomas; Lindblad, Marianne; Nygren, Erik; Raghavan, Sukanya; Sanchez, Joaquin; Stanford, Michael; Sun, Jia-Bin; Svennerholm, Ann-Mari; Tengvall, Sara

    2005-03-15

    Mucosal immunisation may be used both to protect the mucosal surfaces against infections and as a means for immunological treatment of peripheral immunopathological disorders through the induction of systemic antigen-specific tolerance ('oral tolerance'). The development of mucosal vaccines, whether for prevention of infectious diseases or for oral tolerance immunotherapy, requires efficient antigen delivery and adjuvant systems that can help to present the appropriate vaccine or immunotherapy antigens to the mucosal immune system. The most potent (but also toxic) mucosal adjuvants are cholera toxin (CT) and the closely related Escherichia coli heat-labile enterotoxin (LT), and much effort and significant progress have been made recently to generate toxicologically acceptable derivatives of these toxins with retained adjuvant activity. Among these are the non-toxic, recombinantly produced cholera toxin B-subunit (CTB). CTB is a specific protective antigen component of a widely registered oral cholera vaccine as well as a promising vector for either giving rise to mucosal anti-infective immunity or for inducing peripheral anti-inflammatory tolerance to chemically or genetically linked foreign antigens administered mucosally. CT and CTB have also recently been used as combined vectors and adjuvants for markedly promoting ex vivo dendritic cell (DC) vaccination with different antigens and also steering the immune response to the in vivo-reinfused DCs towards either broad Th1 + Th2 + CTL immunity (CT) or Th2 or tolerance (CTB). Another type of mucosal adjuvants is represented by bacterial DNA or synthetic oligodeoxynucleotides containing CpG-motifs, which especially when linked to CTB have been found to effectively stimulate both innate and adaptive mucosal immune responses. The properties and clinical potential of these different classes of adjuvants are being discussed. PMID:15752556

  19. Cholera toxin-like toxin released by Salmonella species in the presence of mitomycin C.

    PubMed

    Molina, N C; Peterson, J W

    1980-10-01

    Several serotypes of Salmonella were shown to release increased amounts of a cholera toxin-like toxin during culture in vitro with mitomycin C (MTC). Filter-sterilized culture supernatants containing the toxin caused elongation of Chinese hamster ovary cells, which could be blocked by heating the supernatants at 100 degrees C for 15 min or by adding mixed gangliosides or monospecific cholera antitoxin. When MTC was not added to the Salmonella cultures, little or no toxin was detected in crude, unconcentrated culture supernatants. Optimal production of toxin was observed in the presence of 0.5 micrograms of MTC per ml in shake flask cultures of Casamino Acids-yeast extract medium, Syncase, or peptone saline at 37 degrees C. Meat infusion media (heart infusion and brain heart infusion) plus MTC resulted in poor toxin yield. Culture filtrates frequently could be diluted 1:8 and still result in elongation of Chinese hamster ovary cells. PMID:7002788

  20. Fate and lability of silver in soils: Effect of ageing

    EPA Science Inventory

    The fate and lability of added soluble Ag in soils over time was examined by measurement of labile metal (E-value) by isotopic dilution using the 110mAg radioactive isotope and the solid-phase speciation of Ag by X-ray absorption near edge structure (XANES) spectrosco...

  1. Identification of labile Zn sites in drug-target proteins.

    PubMed

    Lee, Yu-Ming; Wang, Yi-Ting; Duh, Yulander; Yuan, Hanna S; Lim, Carmay

    2013-09-25

    Labile Zn fingers (Zfs) in proteins contain Zn-bound thiolates that can react with electrophilic agents, causing Zn(2+) ejection and protein unfolding. Such labile Zfs have been shown to be Cys4 or Cys3His cores whose Zn-bound Cys have no hydrogen bonds. Our aim here is to identify labile Zfs in proteins that are promising drug targets using these features. To prove the strategy used, we showed that five proteins with predicted labile Zfs reacted with Zn-ejecting agents, whereas five proteins with no or inert Zfs did not. The comprehensive set of labile Zfs provides new drug targets and guidelines to redesign Zn-ejecting compounds with improved specificity. PMID:24010488

  2. *CYANOBACTERIA AND THEIR TOXINS

    EPA Science Inventory

    Cyanobacteria, or blue-green algae, are naturally-occurring contaminants of surface waters worldwide. These photosynthesizing prokaryotes thrive in warm, shallow, nutrient-rich waters. Many produce potent toxins as secondary metabolites. Cyanobacteria toxins have been document...

  3. Detection of Protein Toxins

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We have focused on ricin, shiga-like toxin, botulinum neurotoxin (BoNT), and staphylococcal enterotoxin A (SEA), developing sensitive test methods for toxins and marker compounds in food matrices. Although animal models provide the best means for risk assessment, especially for crude toxins in compl...

  4. [Intoxication of botulinum toxin].

    PubMed

    Chudzicka, Aleksandra

    2015-09-01

    Botulinum toxin is an egzotoxin produced by Gram positive bacteria Clostridium botulinum. It is among the most potent toxins known. The 3 main clinical presentations of botulism are as follows: foodborne botulism, infant botulism and wound botulism. The main symptom of intoxication is flat muscles paralysis. The treatment is supportive care and administration of antitoxin. In prevention the correct preparing of canned food is most important. Botulinum toxin is accepted as a biological weapon. PMID:26449577

  5. Dietary vitamin E reduces labile iron in rat tissues.

    PubMed

    Ibrahim, Wissam; Chow, Ching Kuang

    2005-01-01

    Previous studies have shown that dietary vitamin E reduced generation and/or levels of superoxide. As superoxide has potential to release iron from its transport and storage proteins, and labile or available form of iron is capable of catalyzing the formation of reactive hydroxyl radicals, the effect of dietary vitamin E on labile iron pool was studied in rats. One-month-old Sprague-Dawley male and female rats were fed a basal vitamin E-deficient diet supplemented with 0, 20, 200, or 2,000 IU vitamin E/kg diet for 90 days. The levels of labile iron were measured in the liver, kidney, spleen, heart and skeletal muscle. Additionally, the levels of lipid peroxidation products were measured. The results showed that, except for labile iron in the heart of male rats, dietary vitamin E dose dependently reduced the levels of labile iron and lipid peroxidation products in all tissues of male and female rats. The findings suggest that dietary vitamin E may protect against oxidative tissue damage by reducing the generation and/or level of superoxide, which in turn attenuates the release of iron from its protein complexes. PMID:16292753

  6. Nanoparticle-detained toxins for safe and effective vaccination

    NASA Astrophysics Data System (ADS)

    Hu, Che-Ming J.; Fang, Ronnie H.; Luk, Brian T.; Zhang, Liangfang

    2013-12-01

    Toxoid vaccines--vaccines based on inactivated bacterial toxins--are routinely used to promote antitoxin immunity for the treatment and prevention of bacterial infections. Following chemical or heat denaturation, inactivated toxins can be administered to mount toxin-specific immune responses. However, retaining faithful antigenic presentation while removing toxin virulence remains a major challenge and presents a trade-off between efficacy and safety in toxoid development. Here, we show a nanoparticle-based toxin-detainment strategy that safely delivers non-disrupted pore-forming toxins for immune processing. Using erythrocyte membrane-coated nanoparticles and staphylococcal α-haemolysin, we demonstrate effective virulence neutralization via spontaneous particle entrapment. Compared with vaccination with heat-denatured toxin, mice vaccinated with the nanoparticle-detained toxin showed superior protective immunity against toxin-mediated adverse effects. We find that the non-disruptive detoxification approach benefited the immunogenicity and efficacy of toxoid vaccines. We anticipate that this study will open new possibilities in the preparation of antitoxin vaccines against the many virulence factors that threaten public health.

  7. Botulinum toxin injection - larynx

    MedlinePLUS

    Injection laryngoplasty; Botox-larynx: spasmodic dysphonia-BTX; Essential voice tremor (EVT)-btx; Glottic insufficiency; Percutaneous electromyography-guided botulinum toxin treatment; Percutaneous indirect laryngoscopy- ...

  8. [Botulinum toxins for pain].

    PubMed

    Soinila, Seppo; Haanp, Maija

    2011-01-01

    We review the evidence of botulinum toxins in the treatment of pain. Main indications of botulinum toxin treatment, dystonia and spasticity, involve pain. Increasing evidence suggests direct analgesic effects of botulinum. Botulinum inhibits release of pain mediators (substance P, CGRP, excitatory amino acids, ATP, noradrenaline). Clinical trials have consistently shown analgesic effect of botulinum toxin in post-stroke shoulder pain, bladder dysfunction, chronic migraine, neuropathic pain, bruxism and lateral epicondylitis. Other pain conditions have been studied with yet uncertain results. It seems that the number of patients who would benefit from botulinum toxin treatment will increase considerably in the future. PMID:22238920

  9. Heterologous Expression of Toxins from Bacterial Toxin-Antitoxin Systems in Eukaryotic Cells: Strategies and Applications.

    PubMed

    Yeo, Chew Chieng; Abu Bakar, Fauziah; Chan, Wai Ting; Espinosa, Manuel; Harikrishna, Jennifer Ann

    2016-01-01

    Toxin-antitoxin (TA) systems are found in nearly all prokaryotic genomes and usually consist of a pair of co-transcribed genes, one of which encodes a stable toxin and the other, its cognate labile antitoxin. Certain environmental and physiological cues trigger the degradation of the antitoxin, causing activation of the toxin, leading either to the death or stasis of the host cell. TA systems have a variety of functions in the bacterial cell, including acting as mediators of programmed cell death, the induction of a dormant state known as persistence and the stable maintenance of plasmids and other mobile genetic elements. Some bacterial TA systems are functional when expressed in eukaryotic cells and this has led to several innovative applications, which are the subject of this review. Here, we look at how bacterial TA systems have been utilized for the genetic manipulation of yeasts and other eukaryotes, for the containment of genetically modified organisms, and for the engineering of high expression eukaryotic cell lines. We also examine how TA systems have been adopted as an important tool in developmental biology research for the ablation of specific cells and the potential for utility of TA systems in antiviral and anticancer gene therapies. PMID:26907343

  10. Heterologous Expression of Toxins from Bacterial Toxin-Antitoxin Systems in Eukaryotic Cells: Strategies and Applications

    PubMed Central

    Yeo, Chew Chieng; Abu Bakar, Fauziah; Chan, Wai Ting; Espinosa, Manuel; Harikrishna, Jennifer Ann

    2016-01-01

    Toxin-antitoxin (TA) systems are found in nearly all prokaryotic genomes and usually consist of a pair of co-transcribed genes, one of which encodes a stable toxin and the other, its cognate labile antitoxin. Certain environmental and physiological cues trigger the degradation of the antitoxin, causing activation of the toxin, leading either to the death or stasis of the host cell. TA systems have a variety of functions in the bacterial cell, including acting as mediators of programmed cell death, the induction of a dormant state known as persistence and the stable maintenance of plasmids and other mobile genetic elements. Some bacterial TA systems are functional when expressed in eukaryotic cells and this has led to several innovative applications, which are the subject of this review. Here, we look at how bacterial TA systems have been utilized for the genetic manipulation of yeasts and other eukaryotes, for the containment of genetically modified organisms, and for the engineering of high expression eukaryotic cell lines. We also examine how TA systems have been adopted as an important tool in developmental biology research for the ablation of specific cells and the potential for utility of TA systems in antiviral and anticancer gene therapies. PMID:26907343

  11. Repeated Labilization-Reconsolidation Processes Strengthen Declarative Memory in Humans

    PubMed Central

    Forcato, Cecilia; Rodrguez, Mara L. C.; Pedreira, Mara E.

    2011-01-01

    The idea that memories are immutable after consolidation has been challenged. Several reports have shown that after the presentation of a specific reminder, reactivated old memories become labile and again susceptible to amnesic agents. Such vulnerability diminishes with the progress of time and implies a re-stabilization phase, usually referred to as reconsolidation. To date, the main findings describe the mechanisms associated with the labilization-reconsolidation process, but little is known about its functionality from a biological standpoint. Indeed, two functions have been proposed. One suggests that destabilization of the original memory after the reminder allows the integration of new information into the background of the original memory (memory updating), and the other suggests that the labilization-reconsolidation process strengthens the original memory (memory strengthening). We have previously reported the reconsolidation of human declarative memories, demonstrating memory updating in the framework of reconsolidation. Here we deal with the strengthening function attributed to the reconsolidation process. We triggered labilization-reconsolidation processes successively by repeated presentations of the proper reminder. Participants learned an association between five cue-syllables and their respective response-syllables. Twenty-four hours later, the paired-associate verbal memory was labilized by exposing the subjects to one, two or four reminders. The List-memory was evaluated on Day 3 showing that the memory was improved when at least a second reminder was presented in the time window of the first labilization-reconsolidation process prompted by the earlier reminder. However, the improvement effect was revealed on Day 3, only when at least two reminders were presented on Day2 and not as a consequence of only retrieval. Therefore, we propose central concepts for the reconsolidation process, emphasizing its biological role and the parametrical constrains for this function to be operative. PMID:21850268

  12. Using Stimulants to Treat ADHD-Related Emotional Lability

    PubMed Central

    Posner, Jonathan; Kass, Erica; Hulvershorn, Leslie

    2014-01-01

    Emotional lability, or sudden strong shifts in emotion, commonly occurs in youth with attention-deficit/hyperactivity disorder. Although these symptoms are impairing and disruptive, relatively little research has addressed their treatment, likely due to the difficulty of reliable and valid assessment. Promising signals for symptom improvement have come from recent studies using stimulants in adults, children and adolescents. Similarly, neuroimaging studies have begun to identify neurobiological mechanisms underlying stimulants’ impact on emotion regulation capacities. Here, we review these recent clinical and neuroimaging findings, as well as neurocognitive models for emotional lability in ADHD, issues of relevance to prescribers and the important role of psychiatric comorbidity with treatment choices. PMID:25135778

  13. Labile trace elements in carbonaceous chondrites - A survey

    NASA Technical Reports Server (NTRS)

    Xiao, Xiaoyue; Lipschutz, Michael E.

    1992-01-01

    Data are presented on 14 trace elements, including Co, Au, Ga, Rb, Sb, Ag, Se, Cs, Te, Zn, Cd, Bi, Tl, and In (nearly all of which are moderately or highly labile in meteorites), obtained by radiochemical neutron activation analyses of 42 C2-C6 chondrites, all but three from Antarctica. The data indicate that carbonaceous chondrites of petrographic types 2-6 define compositional continua. It is suggested that carbonaceous C2-C6 chondrites may reflect a mixture of material that formed at low temperatures and that contained cosmic levels of highly labile elements, with material that was devoid of them.

  14. Defense against toxin weapons

    SciTech Connect

    Franz, D.R.

    1994-01-01

    The purpose of this manual is to provide basic information on biological toxins to military leaders and health-care providers at all levels to help them make informed decisions on protecting their troops from toxins. Much of the information contained herein will also be of interest to individuals charged with countering domestic and international terrorism. We typically fear what we do not understand.

  15. Long-Term Sentinel Surveillance for Enterotoxigenic Escherichia coli and Non-O157 Shiga Toxin-Producing E. coli in Minnesota

    PubMed Central

    Medus, Carlota; Besser, John M.; Juni, Billie A.; Koziol, Bonnie; Lappi, Victoria; Smith, Kirk E.; Hedberg, Craig W.

    2016-01-01

    Background. Enterotoxigenic Escherichia coli (ETEC) and non-O157 Shiga toxin-producing E. coli (STEC) are not detected by conventional culture methods. The prevalence of ETEC infections in the United States is unknown, and recognized cases are primarily associated with foreign travel. Gaps remain in our understanding of STEC epidemiology. Methods. Two sentinel surveillance sites were enrolled: an urban health maintenance organization laboratory (Laboratory A) and a rural hospital laboratory (Laboratory B). Residual sorbitol MacConkey (SMAC) plates from stool cultures performed at Laboratory A (1996–2006) and Laboratory B (2000–2008) were collected. Colony sweeps from SMAC plates were tested for genes encoding STEC toxins stx1 and stx2 (1996–2008) and ETEC heat-labile and heat-stable toxins eltB, estA 1, 2 and 3 (2000–2008) by polymerase chain reaction (PCR)-based assays. Results. In Laboratory A, a bacterial pathogen was identified in 7.0% of 21 970 specimens. During 1996–2006, Campylobacter was the most common bacterial pathogen (2.7% of cultures), followed by Salmonella (1.2%), Shigella (1.0%), and STEC (0.9%). Among STEC (n = 196), O157 was the most common serogroup (31%). During 2000–2006, ETEC (1.9%) was the second most common bacterial pathogen after Campylobacter (2.6%). In Laboratory B, of 19 293 specimens tested, a bacterial pathogen was identified for 5.5%, including Campylobacter (2.1%), STEC (1.3%), Salmonella (1.0%), and ETEC (0.8%). Among STEC (n = 253), O157 was the leading serogroup (35%). Among ETEC cases, 61% traveled internationally. Conclusions. Enterotoxigenic E. coli and STEC infections were as common as most other enteric bacterial pathogens, and ETEC may be detected more frequently by culture-independent multiplex PCR diagnostic methods. A high proportion of ETEC cases were domestically acquired. PMID:26913288

  16. Neuropsychological Correlates of Emotional Lability in Children with ADHD

    ERIC Educational Resources Information Center

    Banaschewski, Tobias; Jennen-Steinmetz, Christine; Brandeis, Daniel; Buitelaar, Jan K.; Kuntsi, Jonna; Poustka, Luise; Sergeant, Joseph A.; Sonuga-Barke, Edmund J.; Frazier-Wood, Alexis C.; Albrecht, Bjorn; Chen, Wai; Uebel, Henrik; Schlotz, Wolff; van der Meere, Jaap J.; Gill, Michael; Manor, Iris; Miranda, Ana; Mulas, Fernando; Oades, Robert D.; Roeyers, Herbert; Rothenberger, Aribert; Steinhausen, Hans-Christoph; Faraone, Stephen V.; Asherson, Philip

    2012-01-01

    Background: Emotional lability (EL) is commonly seen in patients with attention-deficit/hyperactivity disorder (ADHD). The reasons for this association remain currently unknown. To address this question, we examined the relationship between ADHD and EL symptoms, and performance on a range of neuropsychological tasks to clarify whether EL symptoms

  17. Neuropsychological Correlates of Emotional Lability in Children with ADHD

    ERIC Educational Resources Information Center

    Banaschewski, Tobias; Jennen-Steinmetz, Christine; Brandeis, Daniel; Buitelaar, Jan K.; Kuntsi, Jonna; Poustka, Luise; Sergeant, Joseph A.; Sonuga-Barke, Edmund J.; Frazier-Wood, Alexis C.; Albrecht, Bjorn; Chen, Wai; Uebel, Henrik; Schlotz, Wolff; van der Meere, Jaap J.; Gill, Michael; Manor, Iris; Miranda, Ana; Mulas, Fernando; Oades, Robert D.; Roeyers, Herbert; Rothenberger, Aribert; Steinhausen, Hans-Christoph; Faraone, Stephen V.; Asherson, Philip

    2012-01-01

    Background: Emotional lability (EL) is commonly seen in patients with attention-deficit/hyperactivity disorder (ADHD). The reasons for this association remain currently unknown. To address this question, we examined the relationship between ADHD and EL symptoms, and performance on a range of neuropsychological tasks to clarify whether EL symptoms…

  18. How to Compute Labile Metal-Ligand Equilibria

    ERIC Educational Resources Information Center

    de Levie, Robert

    2007-01-01

    The different methods used for computing labile metal-ligand complexes, which are suitable for an iterative computer solution, are illustrated. The ligand function has allowed students to relegate otherwise tedious iterations to a computer, while retaining complete control over what is calculated.

  19. Activation of the NLRP3 inflammasome by cellular labile iron.

    PubMed

    Nakamura, Kyohei; Kawakami, Toru; Yamamoto, Naoki; Tomizawa, Miyu; Fujiwara, Tohru; Ishii, Tomonori; Harigae, Hideo; Ogasawara, Kouetsu

    2016-02-01

    Cellular labile iron, which contains chelatable redox-active Fe(2+), has been implicated in iron-mediated cellular toxicity leading to multiple organ dysfunction. Iron homeostasis is controlled by monocytes/macrophages through their iron recycling and storage capacities. Furthermore, iron sequestration by monocytes/macrophages is regulated by pro-inflammatory cytokines including interleukin-1, highlighting the importance of these cells in the crosstalk between inflammation and iron homeostasis. However, a role for cellular labile iron in monocyte/macrophage-mediated inflammatory responses has not been defined. Here we describe how cellular labile iron activates the NLRP3 inflammasome in human monocytes. Stimulation of lipopolysaccharide-primed peripheral blood mononuclear cells with ferric ammonium citrate increases the level of cellular Fe(2+) levels in monocytes and induces production of interleukin-1? in a dose-dependent manner. This ferric ammonium citrate-induced interleukin-1? production is dependent on caspase-1 and is significantly inhibited by an Fe(2+)-specific chelator. Ferric ammonium citrate consistently induced interleukin-1? secretion in THP1 cells, but not in NLRP3-deficient THP1 cells, indicating a requirement for the NLRP3 inflammasome. Additionally, activation of the inflammasome is mediated by potassium efflux, reactive oxygen species-mediated mitochondrial dysfunction, and lysosomal membrane permeabilization. Thus, these results suggest that monocytes/macrophages not only sequestrate iron during inflammation, but also mediate inflammation in response to cellular labile iron, which provides novel insights into the role of iron in chronic inflammation. PMID:26577567

  20. Memory expression is independent of memory labilization/reconsolidation.

    PubMed

    Barreiro, Karina A; Surez, Luis D; Lynch, Victoria M; Molina, Vctor A; Delorenzi, Alejandro

    2013-11-01

    There is growing evidence that certain reactivation conditions restrict the onset of both the destabilization phase and the restabilization process or reconsolidation. However, it is not yet clear how changes in memory expression during the retrieval experience can influence the emergence of the labilization/reconsolidation process. To address this issue, we used the context-signal memory model of Chasmagnathus. In this paradigm a short reminder that does not include reinforcement allows us to evaluate memory labilization and reconsolidation, whereas a short but reinforced reminder restricts the onset of such a process. The current study investigated the effects of the glutamate antagonists, APV (0.6 or 1.5 ?g/g) and CNQX (1 ?g/g), prior to the reminder session on both behavioral expression and the reconsolidation process. Under conditions where the reminder does not initiate the labilization/reconsolidation process, APV prevented memory expression without affecting long-term memory retention. In contrast, APV induced amnesic effects in the long-term when administered before a reminder session that triggers reconsolidation. Under the present parametric conditions, the administration of CNQX prior to the reminder that allows memory to enter reconsolidation impairs this process without disrupting memory expression. Overall, the present findings suggest that memory reactivation--but not memory expression--is necessary for labilization and reconsolidation. Retrieval and memory expression therefore appear not to be interchangeable concepts. PMID:24149057

  1. [Botulism toxin in practice].

    PubMed

    Durand, A; Serment, G

    2003-07-01

    Botulinum toxins (A and B) are neurotoxins derived from Clostridium botulinum. Clostridium are anaerobic bacteria. C. botulinum produces exotoxins (A to G) with distinct antigenicities. The neurotoxins inhibit the release of the neurotransmitter acetylcholine from the axon terminals of motor neurons. Botulinum toxin is officially used in clinic for the treatment of muscular hyperactivity (strabismus, blepharospam, cervical dystonia). Botulinum toxins are also used in non recognized clinical applications: neurogenic incontinence, palmar and axillary hyperhidrosis, chronic anal fissure. The respective formulations of Botox, Dysport and Neurobloc are described. Special considerations for administration are introduced. PMID:12928147

  2. Microwave heating inactivates Shiga Toxin (Stx2) in reconstituted fat-free Milk and adversely affects the nutritional value of cell culture medium

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Microwave exposure is a convenient and widely used method for defrosting, heating, and cooking numerous foods. Microwave cooking is also reported to kill pathogenic microorganisms that often contaminate food. Microwaves act by causing polar molecules in food, such as water, to rapidly rotate, thus...

  3. The ζ Toxin Induces a Set of Protective Responses and Dormancy

    PubMed Central

    Tabone, Mariangela; Gonzalez-Pastor, José E.; Daugelavicius, Rimantas; Ayora, Silvia; Alonso, Juan C.

    2012-01-01

    The ζε module consists of a labile antitoxin protein, ε, which in dimer form (ε2) interferes with the action of the long-living monomeric ζ phosphotransferase toxin through protein complex formation. Toxin ζ, which inhibits cell wall biosynthesis and may be bactericide in nature, at or near physiological concentrations induces reversible cessation of Bacillus subtilis proliferation (protective dormancy) by targeting essential metabolic functions followed by propidium iodide (PI) staining in a fraction (20–30%) of the population and selects a subpopulation of cells that exhibit non-inheritable tolerance (1–5×10−5). Early after induction ζ toxin alters the expression of ∼78 genes, with the up-regulation of relA among them. RelA contributes to enforce toxin-induced dormancy. At later times, free active ζ decreases synthesis of macromolecules and releases intracellular K+. We propose that ζ toxin induces reversible protective dormancy and permeation to PI, and expression of ε2 antitoxin reverses these effects. At later times, toxin expression is followed by death of a small fraction (∼10%) of PI stained cells that exited earlier or did not enter into the dormant state. Recovery from stress leads to de novo synthesis of ε2 antitoxin, which blocks ATP binding by ζ toxin, thereby inhibiting its phosphotransferase activity. PMID:22295078

  4. Staphylococcus aureus toxins.

    PubMed

    Otto, Michael

    2014-02-01

    Staphylococcus aureus is a dangerous pathogen that causes a variety of severe diseases. The virulence of S. aureus is defined by a large repertoire of virulence factors, among which secreted toxins play a preeminent role. Many S. aureus toxins damage biological membranes, leading to cell death. In particular, S. aureus produces potent hemolysins and leukotoxins. Among the latter, some were recently identified to lyse neutrophils after ingestion, representing an especially powerful weapon against bacterial elimination by innate host defense. Furthermore, S. aureus secretes many factors that inhibit the complement cascade or prevent recognition by host defenses. Several further toxins add to this multi-faceted program of S. aureus to evade elimination in the host. This review will give an overview over S. aureus toxins focusing on recent advances in our understanding of how leukotoxins work in receptor-mediated or receptor-independent fashions. PMID:24581690

  5. Botulinum Toxin Therapy

    MedlinePLUS

    ... therapy Overview Before treatment: This woman disliked her deep frown lines. Botulinum toxin therapy: Overview Also called ... you look in the mirror, do you see deep frown lines? These lines bother many people. To ...

  6. A Novel Mode of Translocation for Cytolethal Distending Toxin

    PubMed Central

    Guerra, Lina; Nemec, Kathleen N.; Massey, Shane; Tatulian, Suren A.; Thelestam, Monica; Frisan, Teresa; Teter, Ken

    2008-01-01

    Summary Thermal instability in the toxin catalytic subunit may be a common property of toxins that exit the endoplasmic reticulum (ER) by exploiting the mechanism of ER-associated degradation (ERAD). The Haemophilus ducreyi cytolethal distending toxin (HdCDT) does not utilize ERAD to exit the ER, so we predicted the structural properties of its catalytic subunit (HdCdtB) would differ from other ER-translocating toxins. Here, we document the heat-stable properties of HdCdtB which distinguish it from other ER-translocating toxins. Cell-based assays further suggested that HdCdtB does not unfold before exiting the ER and that it may move directly from the ER lumen to the nucleoplasm. These observations suggest a novel mode of ER exit for HdCdtB. PMID:19118582

  7. Photo-lability of deep ocean dissolved black carbon

    NASA Astrophysics Data System (ADS)

    Stubbins, A.; Niggemann, J.; Dittmar, T.

    2012-01-01

    Dissolved black carbon (DBC), defined here as condensed aromatics isolated from seawater via PPL solid phase extraction and quantified as benzene polycarboxylic acid oxidation products, is a significant component of the oceanic dissolved organic carbon (DOC) pool. These condensed aromatics are widely distributed in the open ocean and appear to be tens of thousands of years old. As such DBC is regarded as highly refractory. In the current study, the photo-lability of DBC, DOC and coloured dissolved organic matter (CDOM; ultraviolet-visible absorbance) were determined over the course of a 28 d irradiation of North Atlantic Deep Water under a solar simulator. During the irradiation DBC fell from 1044 164 nM C to 55 15 nM C, a 20-fold decrease in concentration. Dissolved black carbon photo-degradation was more rapid and more extensive than for bulk CDOM and DOC. Further, the photo-lability of components of the DBC pool increased with their degree of aromatic condensation. These trends indicate that a continuum of compounds of varying photo-lability exists within the marine DOC pool. In this continuum, photo-lability scales with aromatic character, specifically the degree of condensation. Scaling the rapid photo-degradation of DBC to rates of DOC photo-mineralisation for the global ocean leads to an estimated photo-chemical half-life for oceanic DBC of less than 800 yr. This is more than an order of magnitude shorter than the apparent age of DBC in the ocean. Photo-degradation is therefore posited as the primary sink for oceanic DBC and the survival of DBC molecules in the oceans for millennia appears to be facilitated not by their inherent inertness but by the rate at which they are cycled through the surface ocean's photic zone.

  8. Evaluation of "labile" metal in sediments by anodic stripping voltammetry.

    PubMed

    Waller, P A; Pickering, W F

    1990-10-01

    A procedure for determining the "labile" metal content of contaminated sediments (in different chemical environments) has been critically examined. The sediments were extracted overnight with different chemical solutions and the suspensions were analysed by differential pulse anodic stripping voltammetry. The extractants used have been recommended for soil/sediment speciation schemes, and by examination of the suspensions directly in the ASV cell, errors due to re-adsorption of released metal ion were minimized. The existence of different chemical forms of metal was signified by changes in peak shape and position or by the appearance of additional peaks. With complexing agents present the peak size was pH-dependent. The limitations of the ASV/suspension analysis technique have been carefully evaluated by using ten different extractants and seventeen sediments. The "lability" results obtained have been compared with the values obtained from a cation-exchanger transfer procedure. For characterizing the lability behaviour of the metal contents of sediments, preliminary extraction into a minimum of four base solutions is advisable, e.g., 0.02M nitric acid (low-pH labile); hydroxylamine in acetic acid (reducing conditions); acetic acid/acetate buffer (weakly sorbed and carbonate-bound) and 0.05M calcium chloride (exchangeable fraction at natural system pH), where the terms in parentheses describe the character of the fraction. The results are critically compared with those obtained by atomic-absorption analysis of the extracts and with those obtained by an earlier ion-exchange fractionation scheme. The advantages and limitations of the ASY systems are discussed. PMID:18965053

  9. [Toxins of Clostridium perfringens as a natural and bioterroristic threats].

    PubMed

    Omernik, Andrzej; Płusa, Tadeusz

    2015-09-01

    Clostridium perfringens is absolutely anaerobic rod-shaped, sporeforming bacterium. The morbidity is connected with producing toxins. Depending on the type of toxin produced Clostridium perfringens can be divided into five serotypes:A-E. Under natural conditions, this bacterium is responsible for local outbreaks of food poisoning associated with eating contaminated food which which was improperly heat treated. Some countries with lower economic level are endemic foci of necrotizing enteritis caused by Clostridium perfringens. The bacterium is also a major cause of gas gangrene. It is a disease, associated with wound infection, with potentially fatal prognosis in the case of treatment's delays. In the absence of early radical surgery, antibiotic therapy and (if available) hyperbaric treatment leads to the spread of toxins in the body causing shock, coma and death. Due to the force of produced toxins is a pathogen that poses a substrate for the production of biological weapons. It could potentially be used to induce outbreaks of food poisoning and by missiles contamination by spore lead to increased morbidity of gas gangrene in injured soldiers. C. perfringens types B and D produce epsilon toxin considered to be the third most powerful bacterial toxin. Because of the ability to disperse the toxin as an aerosol and a lack of methods of treatment and prevention of poisoning possible factors it is a potential tool for bioterrorism It is advisable to continue research into vaccines and treatments for poisoning toxins of C. perfringens. PMID:26449576

  10. Labile disulfide bonds are common at the leucocyte cell surface

    PubMed Central

    Metcalfe, Clive; Cresswell, Peter; Ciaccia, Laura; Thomas, Benjamin; Barclay, A. Neil

    2011-01-01

    Redox conditions change in events such as immune and platelet activation, and during viral infection, but the biochemical consequences are not well characterized. There is evidence that some disulfide bonds in membrane proteins are labile while others that are probably structurally important are not exposed at the protein surface. We have developed a proteomic/mass spectrometry method to screen for and identify non-structural, redox-labile disulfide bonds in leucocyte cell-surface proteins. These labile disulfide bonds are common, with several classes of proteins being identified and around 30 membrane proteins regularly identified under different reducing conditions including using enzymes such as thioredoxin. The proteins identified include integrins, receptors, transporters and cellcell recognition proteins. In many cases, at least one cysteine residue was identified by mass spectrometry as being modified by the reduction process. In some cases, functional changes are predicted (e.g. in integrins and cytokine receptors) but the scale of molecular changes in membrane proteins observed suggests that widespread effects are likely on many different types of proteins including enzymes, adhesion proteins and transporters. The results imply that membrane protein activity is being modulated by a redox regulator mechanism. PMID:22645650

  11. Copper lability in soils subjected to intermittent submergence.

    PubMed

    Biasioli, M; Kirby, J K; Hettiarachchi, G M; Ajmone-Marsan, F; McLaughlin, M J

    2010-01-01

    Reducing conditions in soils can have significant influences on the availability of nutrient and toxic metals, through their remobilization, their release through reductive dissolution of oxide phases, and from the formation of precipitates. In the literature, contrasting results are reported on the effects of temporary waterlogging conditions on the availability of metals. In the present study, changes in the "labile" or "potentially available" pool of copper (Cu) in soils as a consequence of up to three intermittent soil submergence cycles was investigated using isotopic dilution. The soils (an Oxisol and an Inceprisol) selected were amended in the field with both biosolids-Cu and salt-Cu. Intermittent soil submergence was found to have a significant effect on the lability of Cu in soils, with E(total) values generally increasing in all the treatments with the different submergence cycles, the highest lability of Cu observed in the Cu-salt treatment. The presence of nonexchangeable colloidal forms of Cu, influenced by treatments and submergence cycles, was also reported. PMID:21284302

  12. Targeted Silencing of Anthrax Toxin Receptors Protects against Anthrax Toxins*

    PubMed Central

    Arvalo, Maria T.; Navarro, Ashley; Arico, Chenoa D.; Li, Junwei; Alkhatib, Omar; Chen, Shan; Diaz-Arvalo, Diana; Zeng, Mingtao

    2014-01-01

    Anthrax spores can be aerosolized and dispersed as a bioweapon. Current postexposure treatments are inadequate at later stages of infection, when high levels of anthrax toxins are present. Anthrax toxins enter cells via two identified anthrax toxin receptors: tumor endothelial marker 8 (TEM8) and capillary morphogenesis protein 2 (CMG2). We hypothesized that host cells would be protected from anthrax toxins if anthrax toxin receptor expression was effectively silenced using RNA interference (RNAi) technology. Thus, anthrax toxin receptors in mouse and human macrophages were silenced using targeted siRNAs or blocked with specific antibody prior to challenge with anthrax lethal toxin. Viability assays were used to assess protection in macrophages treated with specific siRNA or antibody as compared with untreated cells. Silencing CMG2 using targeted siRNAs provided almost complete protection against anthrax lethal toxin-induced cytotoxicity and death in murine and human macrophages. The same results were obtained by prebinding cells with specific antibody prior to treatment with anthrax lethal toxin. In addition, TEM8-targeted siRNAs also offered significant protection against lethal toxin in human macrophage-like cells. Furthermore, silencing CMG2, TEM8, or both receptors in combination was also protective against MEK2 cleavage by lethal toxin or adenylyl cyclase activity by edema toxin in human kidney cells. Thus, anthrax toxin receptor-targeted RNAi has the potential to be developed as a life-saving, postexposure therapy against anthrax. PMID:24742682

  13. Targeted silencing of anthrax toxin receptors protects against anthrax toxins.

    PubMed

    Arvalo, Maria T; Navarro, Ashley; Arico, Chenoa D; Li, Junwei; Alkhatib, Omar; Chen, Shan; Diaz-Arvalo, Diana; Zeng, Mingtao

    2014-05-30

    Anthrax spores can be aerosolized and dispersed as a bioweapon. Current postexposure treatments are inadequate at later stages of infection, when high levels of anthrax toxins are present. Anthrax toxins enter cells via two identified anthrax toxin receptors: tumor endothelial marker 8 (TEM8) and capillary morphogenesis protein 2 (CMG2). We hypothesized that host cells would be protected from anthrax toxins if anthrax toxin receptor expression was effectively silenced using RNA interference (RNAi) technology. Thus, anthrax toxin receptors in mouse and human macrophages were silenced using targeted siRNAs or blocked with specific antibody prior to challenge with anthrax lethal toxin. Viability assays were used to assess protection in macrophages treated with specific siRNA or antibody as compared with untreated cells. Silencing CMG2 using targeted siRNAs provided almost complete protection against anthrax lethal toxin-induced cytotoxicity and death in murine and human macrophages. The same results were obtained by prebinding cells with specific antibody prior to treatment with anthrax lethal toxin. In addition, TEM8-targeted siRNAs also offered significant protection against lethal toxin in human macrophage-like cells. Furthermore, silencing CMG2, TEM8, or both receptors in combination was also protective against MEK2 cleavage by lethal toxin or adenylyl cyclase activity by edema toxin in human kidney cells. Thus, anthrax toxin receptor-targeted RNAi has the potential to be developed as a life-saving, postexposure therapy against anthrax. PMID:24742682

  14. Naturally Occurring Food Toxins

    PubMed Central

    Dolan, Laurie C.; Matulka, Ray A.; Burdock, George A.

    2010-01-01

    Although many foods contain toxins as a naturally-occurring constituent or, are formed as the result of handling or processing, the incidence of adverse reactions to food is relatively low. The low incidence of adverse effects is the result of some pragmatic solutions by the US Food and Drug Administration (FDA) and other regulatory agencies through the creative use of specifications, action levels, tolerances, warning labels and prohibitions. Manufacturers have also played a role by setting limits on certain substances and developing mitigation procedures for process-induced toxins. Regardless of measures taken by regulators and food producers to protect consumers from natural food toxins, consumption of small levels of these materials is unavoidable. Although the risk for toxicity due to consumption of food toxins is fairly low, there is always the possibility of toxicity due to contamination, overconsumption, allergy or an unpredictable idiosyncratic response. The purpose of this review is to provide a toxicological and regulatory overview of some of the toxins present in some commonly consumed foods, and where possible, discuss the steps that have been taken to reduce consumer exposure, many of which are possible because of the unique process of food regulation in the United States. PMID:22069686

  15. Marine Toxins: An Overview

    NASA Astrophysics Data System (ADS)

    Fusetani, Nobuhiro

    Oceans provide enormous and diverse space for marine life. Invertebrates are conspicuous inhabitants in certain zones such as the intertidal; many are soft-bodied, relatively immobile and lack obvious physical defenses. These animals frequently have evolved chemical defenses against predators and overgrowth by fouling organisms. Marine animals may accumulate and use a variety of toxins from prey organisms and from symbiotic microorganisms for their own purposes. Thus, toxic animals are particularly abundant in the oceans. The toxins vary from small molecules to high molecular weight proteins and display unique chemical and biological features of scientific interest. Many of these substances can serve as useful research tools or molecular models for the design of new drugs and pesticides. This chapter provides an initial survey of these toxins and their salient properties.

  16. [Toxins as a biological weapon].

    PubMed

    P?usa, Tadeusz

    2015-09-01

    The criteria for recognizing a chemical compound for the toxin are vague and gave it the possibility of inclusion in this group a number of biological agents. Toxins list is extensive, but the interest is focused on bacterial toxins, poisons derived from snake venoms, algae and plant proteins, and small molecules. Particular attention is focused on the so-called "sea" toxins, which include tetrodotoxin, brevetoxin and saxitoxin. This indicates the search for a new hitherto unknown potential bioterrorist threats. PMID:26449572

  17. The association of emotional lability and emotional and behavioral difficulties among children with and without ADHD.

    PubMed

    Rosen, Paul J; Walerius, Danielle M; Fogleman, Nicholas D; Factor, Perry I

    2015-12-01

    Children with ADHD often demonstrate a pattern of emotional lability characterized by sudden and intense shifts in affect. Emotional lability has been linked to emotional and behavioral problems in children with and without ADHD, but few studies have examined emotional lability over time. This study examined the effects of emotional lability over time on the behavioral and emotional difficulties of children with and without ADHD using an ecological momentary assessment (EMA) methodology. One hundred and two children aged 8-12years (56 with ADHD and 46 without ADHD) and their parents completed baseline measures of the children's behavioral and emotional difficulties. Parents then completed a 28-day 3-times daily EMA assessment protocol to rate their child's emotional lability. Results suggested that emotional lability was associated with internalizing and/or externalizing diagnoses independent of ADHD diagnostic status, but was not directly associated with ADHD. Hierarchical regression analyses supported ADHD diagnostic status as a moderator of the association of greater EMA-derived emotional lability with children's behavioral difficulties, such that greater emotional lability was associated with greater behavioral difficulties among children with ADHD but not among children without ADHD. Results indicated that greater emotional lability was directly linked with greater emotional difficulties and that this relation was not moderated by ADHD diagnostic status. Overall, this study suggested that emotional lability is related to emotional difficulties independent of ADHD, but is differentially related to behavioral difficulties among children with and without ADHD. PMID:25957599

  18. Bacterial toxins--an overview.

    PubMed

    Lahiri, S S

    2000-11-01

    Toxins are non-replicating agents of biological origin. They are non-infectious, non-contagious, and non-curable by antibiotics or chemotherapeutic agents. However, individuals can be protected by vaccination. The multifactorial nature of virulence of toxin and toxin producers, produces comparative and cooperative pathogenesis, and this makes studies all the more difficult. Antibody raised against all components helps in this pursuit. The toxins have been classified into seven different classes and over 44 bacterial toxins have been discussed. The botulinum toxin is by far the most toxic substance in the world. All the toxins produced are either secreted out, called exotoxins (proteins), or are entrapped in the cell membrane, called endotoxins (lipopolysaccharides). These toxins are di-chain molecules, internalized into the cell by receptor mediated endocytosis, and ADP-ribosylation is the most common mode of action. The toxins produced by bacteria are enterotoxins, neurotoxins, cytotoxins, lysins (e.g., hemolysin), gangrene producing toxins. However, a single bacteria often produces more than one toxin. Bacterial toxins, which are primarily harmful, are also being used for the cure of cancer, killing of mosquito larvae, understanding of basic sciences like ADP-ribosylation, etc. PMID:11126516

  19. Paralytic shellfish toxins inhibit copper uptake in Chlamydomonas reinhardtii.

    PubMed

    Cusick, Kathleen D; Wetzel, Randall K; Minkin, Steven C; Dodani, Sheel C; Wilhelm, Steven W; Sayler, Gary S

    2013-06-01

    Paralytic shellfish toxins are secondary metabolites produced by several species of dinoflagellates and cyanobacteria. Known targets of these toxins, which typically occur at detrimental concentrations during harmful algal blooms, include voltage-gated ion channels in humans and other mammals. However, the effects of the toxins on the co-occurring phytoplankton community remain unknown. The present study examined the molecular mechanisms of the model photosynthetic alga Chlamydomonas reinhardtii in response to saxitoxin exposure as a means of gaining insight into the phytoplankton community response to a bloom. Previous work with yeast indicated that saxitoxin inhibited copper uptake, so experiments were designed to examine whether saxitoxin exhibited a similar mode of action in algae. Expression profiling following exposure to saxitoxin or a copper chelator produced similar profiles in copper homeostasis genes, notably induction of the cytochrome c6 (CYC6) and copper transporter (COPT1, CTR1) genes. Cytochrome c6 is used as an alternative to plastocyanin under conditions of copper deficiency, and immunofluorescence data showed this protein to be present in a significantly greater proportion of saxitoxin-exposed cells compared to controls. Live-cell imaging with a copper-sensor probe for intracellular labile Cu(I) confirmed that saxitoxin blocked copper uptake. Extrapolations of these data to phytoplankton metabolic processes along with the copper transporter as a molecular target of saxitoxin based on existing structural models are discussed. PMID:23423950

  20. Do Vermont's Floodplains Constitute an Important Source of Labile Carbon?

    NASA Astrophysics Data System (ADS)

    Perdrial, J. N.; Dolan, A.; Kemsley, M.

    2014-12-01

    Floodplains are extremely heterogeneous landscapes with respect to soil and sediment composition and can present an important source of carbon (C) during floods. For example, stream bank soils and sediments are zones of active erosion and deposition of sediment associated C. Due to the presence of plants, riparian soils contain high amounts of C that is exchanged between stream waters and banks. Abandoned channels and meander wetlands that remain hydrologically connected to the main channel contain high amounts of organic matter that can be flushed into the stream during high discharge. This heterogeneity, result of floodplain geomorphology, land cover and use, can profoundly impact the amount and type of dissolved organic matter (DOM) introduced into streams. In order to assess DOM characteristics leached from heterogeneous floodplain soils, aqueous soil extracts were performed on soil samples representative of different land covers (n=20) at four depths. Extracts were analyzed for dissolved organic C and total dissolved nitrogen with a Shimadzu C analyzer. Colored dissolved organic matter characteristics was measured with the Aqualog Fluorescence Spectrometer and quantified with parallel factor analysis (PARAFAC). Preliminary data from three floodplains in Vermont (Connecticut, Missisquoi and Mad River) show a 3D variability of longitudinal, lateral, and vertical extents on water-extractable, mobile C. Dissolved organic carbon concentrations in meander swamp samples were found up to 9 times higher than in those of soils from agricultural field indicative of an important C source. Although C concentrations in adjacent fields were low, high abundance of labile C (indicated by tryptophan-like fluorescence) in water extracts from fields indicates recent biological production of C. This labile C is easily processed by microbes and transformed to the greenhouse gas CO2. These results provide important information on the contribution and lability of different floodplain areas (banks, riparian areas and meander wetlands) to C export and can help to make predictions for their export during flooding.

  1. The Children's Affective Lability Scale: a psychometric evaluation of reliability.

    PubMed

    Gerson, A C; Gerring, J P; Freund, L; Joshi, P T; Capozzoli, J; Brady, K; Denckla, M B

    1996-12-20

    The Children's Affective Lability Scale (CALS) is a 20-item parent report measure developed to assess affect regulation in children aged 6-16. It was normed with school children in regular education classrooms and with children hospitalized in a psychiatric facility. Internal-consistency reliability, split-half reliability, and two-week test-retest reliability were excellent. Staff interrater reliability in the psychiatric sample was acceptable. Higher CALS scores were observed in an in-patient psychiatric sample than in either an out-patient or a normative sample. A principal components factor analysis yielded two components for the normative sample. PMID:9029668

  2. Photo-lability of deep ocean dissolved black carbon

    NASA Astrophysics Data System (ADS)

    Stubbins, A.; Niggemann, J.; Dittmar, T.

    2012-05-01

    Dissolved black carbon (DBC), defined here as condensed aromatics isolated from seawater via PPL solid phase extraction and quantified as benzenepolycarboxylic acid (BPCA) oxidation products, is a significant component of the oceanic dissolved organic carbon (DOC) pool. These condensed aromatics are widely distributed in the open ocean and appear to be tens of thousands of years old. As such DBC is regarded as highly refractory. In the current study, the photo-lability of DBC, DOC and coloured dissolved organic matter (CDOM; ultraviolet-visible absorbance) were determined over the course of a 28 day irradiation of North Atlantic Deep Water under a solar simulator. During the irradiation DBC fell from 1044 164 nM-C to 55 15 nM-C, a 20-fold decrease in concentration. Dissolved black carbon photo-degradation was more rapid and more extensive than for bulk CDOM and DOC. The concentration of DBC correlated with CDOM absorbance and the quality of DBC indicated by the ratios of different BPCAs correlated with CDOM absorbance spectral slope, suggesting the optical properties of CDOM may provide a proxy for both DBC concentrations and quality in natural waters. Further, the photo-lability of components of the DBC pool increased with their degree of aromatic condensation. These trends indicate that a continuum of compounds of varying photo-lability exists within the marine DOC pool. In this continuum, photo-lability scales with aromatic character, specifically the degree of condensation. Scaling the rapid photo-degradation of DBC to rates of DOC photo-mineralisation for the global ocean leads to an estimated photo-chemical half-life for oceanic DBC of less than 800 years. This is more than an order of magnitude shorter than the apparent age of DBC in the ocean. Consequently, photo-degradation is posited as the primary sink for oceanic DBC and the apparent survival of DBC molecules in the oceans for millennia appears to be facilitated not by their inherent inertness but by the rate at which they are cycled through the surface ocean's photic zone.

  3. CYANOBACTERIA AND THEIR TOXINS.

    EPA Science Inventory

    Science Questions

    Harmful algal blooms (HAB) of cyanobacteria, also known as blue-green algae, have recently become more spatially and temporally prevalent in the US and worldwide. Cyanobacteria and their highly potent toxins are a significant hazard for human health and ...

  4. CYANOBACTERIA AND THEIR TOXINS

    EPA Science Inventory

    Science Questions

    Harmful algal blooms (HAB) of cyanobacteria, also known as blue-green algae, have recently become more spatially and temporally prevalent in the US and worldwide. Cyanobacteria and their highly potent toxins are a significant hazard for human health and ...

  5. An ultrahigh-resolution mass spectrometry index to estimate natural organic matter lability

    PubMed Central

    D'Andrilli, Juliana; Cooper, William T; Foreman, Christine M; Marshall, Alan G

    2015-01-01

    Rationale Determining the chemical constituents of natural organic matter (NOM) by Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FTICRMS) remains the ultimate measure for probing its source material, evolution, and transport; however, lability and the fate of organic matter (OM) in the environment remain controversial. FTICRMS-derived elemental compositions are presented in this study to validate a new interpretative method to determine the extent of NOM lability from various environments. Methods FTICRMS data collected over the last decade from the same 9.4 tesla instrument using negative electrospray ionization at the National High Magnetic Field Laboratory in Tallahassee, Florida, was used to validate the application of a NOM lability index. Solid-phase extraction cartridges were used to isolate the NOM prior to FTICRMS; mass spectral peaks were calibrated internally by commonly identified NOM homologous series, and molecular formulae were determined for NOM composition and lability analysis. Results A molecular lability boundary (MLB) was developed from the FTICRMS molecular data, visualized from van Krevelen diagrams, dividing the data into more and less labile constituents. NOM constituents above the MLB at H/C ≥1.5 correspond to more labile material, whereas NOM constituents below the MLB, H/C <1.5, exhibit less labile, more recalcitrant character. Of all marine, freshwater, and glacial environments considered for this study, glacial ecosystems were calculated to contain the most labile OM. Conclusions The MLB extends our interpretation of FTICRMS NOM molecular data to include a metric of lability, and generally ranked the OM environments from most to least labile as glacial > marine > freshwater. Applying the MLB is useful not only for individual NOM FTICRMS studies, but also provides a lability threshold to compare and contrast molecular data with other FTICRMS instruments that survey NOM from around the world. Copyright © 2015 The Authors. Rapid Communications in Mass Spectrometry published by John Wiley & Sons Ltd. PMID:26563709

  6. Acid-Labile Polyvinylamine Micro- and Nanogel Capsules

    PubMed Central

    Shi, Lianjun; Berkland, Cory

    2008-01-01

    Hollow nanoparticles represent an emerging area of development for the encapsulation of active ingredients. Expanding the capabilities of these nanomaterials will require continued efforts to infill properties such as size control, biodegradability, and environmental responsiveness. Acid-labile poly(N-vinylformamide) (PNVF) nanocapsules were synthesized by free radical polymerization of N-vinylformamide on the surface of silica nanoparticles. Polymerization in the presence of a novel crosslinker that contains an acid-labile ketal facilitated stable etching of silica nanoparticle templates using sodium hydroxide and recovery of degradable PNVF nanocapsules. The formamido side group of PNVF was then hydrolyzed by extended exposure to sodium hydroxide to produce polyvinylamine (PVAm) micro- and nanocapsules. Both capsule types demonstrated an increasing dissolution rate as pH decreased. In addition, PVAm nanocapsules exhibited swelling in proportion to the relative charge density of the PVAm network (a function of the degree of formamide hydrolysis and pH), presumably due to the repulsion of positively charged amino groups within the elastic shell network. The synthetic approaches reported provide methods to endow nanocapsules with key attributes such as size control, pH sensitive degradation, swelling in response to pH, and amine functionality. PMID:18797513

  7. The Labile Side of Iron Supplementation in CKD.

    PubMed

    Slotki, Itzchak; Cabantchik, Zvi Ioav

    2015-11-01

    The practice of intravenous iron supplementation has grown as nephrologists have gradually moved away from the liberal use of erythropoiesis-stimulating agents as the main treatment for the anemia of CKD. This approach, together with the introduction of large-dose iron preparations, raises the future specter of inadvertent iatrogenic iron toxicity. Concerns have been raised in original studies and reviews about cardiac complications and severe infections that result from long-term intravenous iron supplementation. Regarding the iron preparations specifically, even though all the currently available preparations appear to be relatively safe in the short term, little is known regarding their long-term safety. In this review we summarize current knowledge of iron metabolism with an emphasis on the sources and potentially harmful effects of labile iron, highlight the approaches to identifying labile iron in pharmaceutical preparations and body fluids and its potential toxic role as a pathogenic factor in the complications of CKD, and propose methods for its early detection in at-risk patients. PMID:25999405

  8. Diversity of bacterial type II toxinantitoxin systems: a comprehensive search and functional analysis of novel families

    PubMed Central

    Leplae, Raphal; Geeraerts, Damien; Hallez, Rgis; Guglielmini, Julien; Drze, Pierre; Van Melderen, Laurence

    2011-01-01

    Type II toxinantitoxin (TA) systems are generally composed of two genes organized in an operon, encoding a labile antitoxin and a stable toxin. They were first discovered on plasmids where they contribute to plasmid stability by a phenomenon denoted as addiction, and subsequently in bacterial chromosomes. To discover novel families of antitoxins and toxins, we developed a bioinformatics approach based on the guilt by association principle. Extensive experimental validation in Escherichia coli of predicted antitoxins and toxins increased significantly the number of validated systems and defined novel toxin and antitoxin families. Our data suggest that toxin families as well as antitoxin families originate from distinct ancestors that were assembled multiple times during evolution. Toxin and antitoxin families found on plasmids tend to be promiscuous and widespread, indicating that TA systems move through horizontal gene transfer. We propose that due to their addictive properties, TA systems are likely to be maintained in chromosomes even though they do not necessarily confer an advantage to their bacterial hosts. Therefore, addiction might play a major role in the evolutionary success of TA systems both on mobile genetic elements and in bacterial chromosomes. PMID:21422074

  9. Animal toxins and the kidney.

    PubMed

    Sitprija, Visith

    2008-11-01

    Envenomation or poisoning by toxins from animals poses an important health hazard in the tropics. Animal toxins are complex mixtures of proteins, peptides, enzymes and chemicals. These toxins exert their effects through modulation of ion channels and receptors, and via direct enzyme action. Depolarization or hyperpolarization of ion channels--caused by most marine toxins, and some snake and insect venoms--results in neuromuscular symptoms that can be associated with hemodynamic changes. Toxin enzymes, especially proteases and phospholipase A2, initiate inflammatory processes that involve the generation of proinflammatory cytokines and vasoactive mediators, resulting in systemic and renal hemodynamic alterations. Toxin enzymes also have direct effects on erythrocytes, myocytes, blood coagulation factors, vascular endothelium and epithelial cells. As a result, disseminated intravascular coagulation, bleeding diathesis, intravascular hemolysis and rhabdomyolysis are common after exposure to animal toxins. The renal manifestations of animal toxin envenomation, which are usually acute, result mainly from these enzymatic effects. All renal structures can be affected by animal toxins, and tubular necrosis is common. Acute kidney injury is attributed to decreased renal blood flow (associated with intravascular hemolysis or rhabdomyolysis), disseminated intravascular coagulation or direct tubular toxicity. Immunologic mechanisms have a minor role in the pathophysiology of nephropathy caused by animal toxins. PMID:18813235

  10. Toxins and drug discovery.

    PubMed

    Harvey, Alan L

    2014-12-15

    Components from venoms have stimulated many drug discovery projects, with some notable successes. These are briefly reviewed, from captopril to ziconotide. However, there have been many more disappointments on the road from toxin discovery to approval of a new medicine. Drug discovery and development is an inherently risky business, and the main causes of failure during development programmes are outlined in order to highlight steps that might be taken to increase the chances of success with toxin-based drug discovery. These include having a clear focus on unmet therapeutic needs, concentrating on targets that are well-validated in terms of their relevance to the disease in question, making use of phenotypic screening rather than molecular-based assays, and working with development partners with the resources required for the long and expensive development process. PMID:25448391

  11. Method for detecting biological toxins

    SciTech Connect

    Ligler, F.S.; Campbell, J.R.

    1992-01-01

    Biological toxins are indirectly detected by using polymerase chain reaction to amplify unique nucleic acid sequences coding for the toxins or enzymes unique to toxin synthesis. Buffer, primers coding for the unique nucleic acid sequences and an amplifying enzyme are added to a sample suspected of containing the toxin. The mixture is then cycled thermally to exponentially amplify any of these unique nucleic acid sequences present in the sample. The amplified sequences can be detected by various means, including fluorescence. Detection of the amplified sequences is indicative of the presence of toxin in the original sample. By using more than one set of labeled primers, the method can be used to simultaneously detect several toxins in a sample.

  12. Lability of renal papillary tissue composition in the rat.

    PubMed Central

    Atherton, J C

    1978-01-01

    1. The acute effects of (a) a minor operative procedure using ether as the anaesthetic, and (b) the administration of 0.9% saline as a single I.V. injection in the conscious rat, on renal tissue composition were studied in hydropenic and normally hydrated rats. 2. The operative procedure and anaesthesia induced a rapid and transient decrease in papillary osmolality in both hydropenic and normally hydrated animals, the important contributing factor being a significant decrease in urea content. 3. Administration of a small volume of saline caused a rapid decrease in urea content, and an increase in water content. 4. It is concluded that papillary composition is extremely labile, large changes being produced by relatively minor experimental procedures. PMID:624997

  13. The role of labile sulfur compounds in thermochemical sulfate reduction

    NASA Astrophysics Data System (ADS)

    Amrani, Alon; Zhang, Tongwei; Ma, Qisheng; Ellis, Geoffrey S.; Tang, Yongchun

    2008-06-01

    The reduction of sulfate to sulfide coupled with the oxidation of hydrocarbons to carbon dioxide, commonly referred to as thermochemical sulfate reduction (TSR), is an important abiotic alteration process that most commonly occurs in hot carbonate petroleum reservoirs. In the present study we focus on the role that organic labile sulfur compounds play in increasing the rate of TSR. A series of gold-tube hydrous pyrolysis experiments were conducted with n-octane and CaSO4 in the presence of reduced sulfur (e.g. H2S, S, organic S) at temperatures of 330 and 356 C under a constant confining pressure. The in-situ pH was buffered to 3.5 (?6.3 at room temperature) with talc and silica. For comparison, three types of oil with different total S and labile S contents were reacted under similar conditions. The results show that the initial presence of organic or inorganic sulfur compounds increases the rate of TSR. However, organic sulfur compounds, such as 1-pentanethiol or diethyldisulfide, were significantly more effective in increasing the rate of TSR than H2S or elemental sulfur (on a mole S basis). The increase in rate is achieved at relatively low concentrations of 1-pentanethiol, less than 1 wt% of the total n-octane, which is comparable to the concentration of organic S that is common in many oils (?0.3 wt%). We examined several potential reaction mechanisms to explain the observed reactivity of organic LSC. First, the release of H2S from the thermal degradation of thiols was discounted as an important mechanism due to the significantly greater reactivity of thiol compared to an equivalent amount of H2S. Second, we considered the generation of olefines in association with the elimination of H2S during thermal degradation of thiols because olefines are much more reactive than n-alkanes during TSR. In our experiments, olefines increased the rate of TSR, but were less effective than 1-pentanethiol and other organic LSC. Third, the thermal decomposition of organic LSC creates free-radicals that in turn might initiate a radical chain-reaction that creates more reactive species. Experiments involving radical initiators, such as diethyldisulfide and benzyldisulfide, did not show an increase in reactivity compared to 1-pentanethiol. Therefore, we conclude that none of these can sufficiently explain our observations of the initial stages of TSR; they may, however, be important in the later stages. In order to gain greater insight into the potential mechanism for the observed reactivity of these organic sulfur compounds during TSR, we applied density functional theory-based molecular modeling techniques to our system. The results of these calculations indicate that 1-pentanethiol or its thermal degradation products may directly react with sulfate and reduce the activation energy required to rupture the first S-O bond through the formation of a sulfate ester. This study demonstrates the importance of labile sulfur compounds in reducing the onset timing and temperature of TSR. It is therefore essential that labile sulfur concentrations are taken into consideration when trying to make accurate predictions of TSR kinetics and the potential for H2S accumulation in petroleum reservoirs.

  14. Ricin detection: tracking active toxin.

    PubMed

    Bozza, William P; Tolleson, William H; Rosado, Leslie A Rivera; Zhang, Baolin

    2015-01-01

    Ricin is a plant toxin with high bioterrorism potential due to its natural abundance and potency in inducing cell death. Early detection of the active toxin is essential for developing appropriate countermeasures. Here we review concepts for designing ricin detection methods, including mechanism of action of the toxin, advantages and disadvantages of current detection assays, and perspectives on the future development of rapid and reliable methods for detecting ricin in environmental samples. PMID:25481398

  15. Toxin Plasmids of Clostridium perfringens

    PubMed Central

    Li, Jihong; Adams, Vicki; Bannam, Trudi L.; Miyamoto, Kazuaki; Garcia, Jorge P.; Uzal, Francisco A.; Rood, Julian I.

    2013-01-01

    SUMMARY In both humans and animals, Clostridium perfringens is an important cause of histotoxic infections and diseases originating in the intestines, such as enteritis and enterotoxemia. The virulence of this Gram-positive, anaerobic bacterium is heavily dependent upon its prolific toxin-producing ability. Many of the ∼16 toxins produced by C. perfringens are encoded by large plasmids that range in size from ∼45 kb to ∼140 kb. These plasmid-encoded toxins are often closely associated with mobile elements. A C. perfringens strain can carry up to three different toxin plasmids, with a single plasmid carrying up to three distinct toxin genes. Molecular Koch's postulate analyses have established the importance of several plasmid-encoded toxins when C. perfringens disease strains cause enteritis or enterotoxemias. Many toxin plasmids are closely related, suggesting a common evolutionary origin. In particular, most toxin plasmids and some antibiotic resistance plasmids of C. perfringens share an ∼35-kb region containing a Tn916-related conjugation locus named tcp (transfer of clostridial plasmids). This tcp locus can mediate highly efficient conjugative transfer of these toxin or resistance plasmids. For example, conjugative transfer of a toxin plasmid from an infecting strain to C. perfringens normal intestinal flora strains may help to amplify and prolong an infection. Therefore, the presence of toxin genes on conjugative plasmids, particularly in association with insertion sequences that may mobilize these toxin genes, likely provides C. perfringens with considerable virulence plasticity and adaptability when it causes diseases originating in the gastrointestinal tract. PMID:23699255

  16. Elemental composition and functional groups in soil labile organic matter fractions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Labile organic matter fractions are major components involved in nutrient cycle in soil. In this chapter, we examine three labile organic matter fraction: light fraction (LF), humic acid (HA) and fulvic acid (HA) in Alabama cotton soils (ultisol) amended with chemical fertilizer (NH4NO3) and poult...

  17. Knowledge Lability: Within-Person Changes in Parental Knowledge and Their Associations with Adolescent Problem Behavior.

    PubMed

    Lippold, Melissa A; Fosco, Gregory M; Ram, Nilam; Feinberg, Mark E

    2016-02-01

    Higher levels of parental knowledge about youth activities have been associated with lower levels of youth risky behavior. Yet little is known about how parental knowledge fluctuates during early adolescence and how those fluctuations are associated with the development of problem behavior. We use the term lability to describe within-person fluctuations in knowledge over time with higher lability indicating greater fluctuations in knowledge from year-to-year. This longitudinal study of rural adolescents (N = 840) investigated if change in parental knowledge across four waves of data from grades 6 to 8 is characterized by lability, and if greater lability is associated with higher youth substance use, delinquency, and internalizing problems in grade 9. Our models indicated that only some of the variance in parental knowledge was accounted for by developmental trends. The remaining residual variance reflects within-person fluctuations around these trends, lability, and measurement and occasion-specific error. Even controlling for level and developmental trends in knowledge, higher knowledge lability (i.e., more fluctuation) was associated with increased risk for later alcohol and tobacco use, and for girls, higher delinquency and internalizing problems. Our findings suggest that lability in parental knowledge has unique implications for adolescent outcomes. The discussion focuses on mechanisms that may link knowledge lability to substance use. Interventions may be most effective if they teach parents to consistently and predictably decrease knowledge across early adolescence. PMID:26381431

  18. Labile carbon and nitrogen from rhizoplane and surface soils of two perennial grasslands

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In semiarid perennial grasslands biogeochemical processes that drive nutrient dynamics may be more closely related to the quantity of labile SOM than to total SOM. A small ephemeral pool of labile soluble organic matter becomes active after pulse precipitation events. Rhizoplane soil associated with...

  19. Mzm1 Influences a Labile Pool of Mitochondrial Zinc Important for Respiratory Function*

    PubMed Central

    Atkinson, Aaron; Khalimonchuk, Oleh; Smith, Pamela; Sabic, Hana; Eide, David; Winge, Dennis R.

    2010-01-01

    Zinc is essential for function of mitochondria as a cofactor for several matrix zinc metalloproteins. We demonstrate that a labile cationic zinc component of low molecular mass exists in the yeast mitochondrial matrix. This zinc pool is homeostatically regulated in response to the cellular zinc status. This pool of zinc is functionally important because matrix targeting of a cytosolic zinc-binding protein reduces the level of labile zinc and interferes with mitochondrial respiratory function. We identified a series of proteins that modulate the matrix zinc pool, one of which is a novel conserved mitochondrial protein designated Mzm1. Mutant mzm1? cells have reduced total and labile mitochondrial zinc, and these cells are hypersensitive to perturbations of the labile pool. In addition, mzm1? cells have a destabilized cytochrome c reductase (Complex III) without any effects on Complexes IV or V. Thus, we have established that a link exists between Complex III integrity and the labile mitochondrial zinc pool. PMID:20404342

  20. Pertussis toxin expression in Drosophila alters the visual response and blocks eating behaviour.

    PubMed

    Fitch, C L; de Sousa, S M; O'Day, P M; Neubert, T A; Plantilla, C M; Spencer, M; Yarfitz, S; Apte, D; Hurley, J B

    1993-03-01

    Pertussis toxin inactivates certain G-proteins by introducing an ADP-ribose group near the carboxyl-terminus of the alpha-subunit. The major pertussis toxin substrate in Drosophila tissues is Go alpha. We introduced a pertussis toxin gene under control of the hsp70 heat-shock promoter into the Drosophila genome. When heat-shocked, transformed flies produce active pertussis toxin which ADP-ribosylates endogenous Go alpha. Pertussis toxin is expressed in photoreceptors, in the lamina of the eye and in epithelial cells lining the gut. As expected from the absence of Go alpha in photoreceptors, pertussis toxin does not affect the photoreceptor component of the Drosophila visual response. However, it abolishes light on- and off-transients in the electroretinogram. These transients normally arise from the lamina, a tissue where Go alpha transcripts have been detected. Pertussis toxin expression also blocks embryonic development and shortens the lifetime of adult Drosophila. Following heat-shock, transformed adults are active, but they fail to take up nutrients because they stop eating. High energy metabolites are significantly depleted shortly after pertussis toxin expression is induced and the flies die within 48 h. PMID:8499226

  1. In situ, high-resolution imaging of labile phosphorus in sediments of a large eutrophic lake.

    PubMed

    Ding, Shiming; Han, Chao; Wang, Yanping; Yao, Lei; Wang, Yan; Xu, Di; Sun, Qin; Williams, Paul N; Zhang, Chaosheng

    2015-05-01

    Understanding the labile status of phosphorus (P) in sediments is crucial for managing a eutrophic lake, but it is hindered by lacking in situ data particularly on a catchment scale. In this study, we for the first time characterized in situ labile P in sediments with the Zr-oxide diffusive gradients in thin films (Zr-oxide DGT) technique at a two-dimensional (2D), submillimeter resolution in a large eutrophic lake (Lake Taihu, China, with an area of 2338km(2)). The concentration of DGT-labile P in the sediment profiles showed strong variation mostly ranging from 0.01 to 0.35mgL(-1) with a considerable number of hotspots. The horizontal heterogeneity index of labile P varied from 0.04 to 4.5. High values appeared at the depths of 0-30mm, likely reflecting an active layer of labile P under the sediment-water interface (SWI). Concentration gradients of labile P were observed from the high-resolution 1D DGT profiles in both the sediment and overlying water layers close to the SWI. The apparent diffusion flux of P across the SWI was calculated between -21 and 65ngcm(-2)d(-1), which showed that the sediments tended to be a source and sink of overlying water P in the algal- and macrophyte-dominated regions, respectively. The DGT-labile P in the 0-30mm active layer showed a better correlation with overlying water P than the labile P measured by ex situ chemical extraction methods. It implies that in situ, high-resolution profiling of labile P with DGT is a more reliable approach and will significantly extend our ability in in situ monitoring of the labile status of P in sediments in the field. PMID:25720671

  2. Ratcheting up protein translocation with anthrax toxin.

    PubMed

    Feld, Geoffrey K; Brown, Michael J; Krantz, Bryan A

    2012-05-01

    Energy-consuming nanomachines catalyze the directed movement of biopolymers in the cell. They are found both dissolved in the aqueous cytosol as well as embedded in lipid bilayers. Inquiries into the molecular mechanism of nanomachine-catalyzed biopolymer transport have revealed that these machines are equipped with molecular parts, including adjustable clamps, levers, and adaptors, which interact favorably with substrate polypeptides. Biological nanomachines that catalyze protein transport, known as translocases, often require that their substrate proteins unfold before translocation. An unstructured protein chain is likely entropically challenging to bind, push, or pull in a directional manner, especially in a way that produces an unfolding force. A number of ingenious solutions to this problem are now evident in the anthrax toxin system, a model used to study protein translocation. Here we highlight molecular ratchets and current research on anthrax toxin translocation. A picture is emerging of proton-gradient-driven anthrax toxin translocation, and its associated ratchet mechanism likely applies broadly to other systems. We suggest a cyclical thermodynamic order-to-disorder mechanism (akin to a heat-engine cycle) is central to underlying protein translocation: peptide substrates nonspecifically bind to molecular clamps, which possess adjustable affinities; polypeptide substrates compress into helical structures; these clamps undergo proton-gated switching; and the substrate subsequently expands regaining its unfolded state conformational entropy upon translocation. PMID:22374876

  3. Ratcheting up protein translocation with anthrax toxin

    PubMed Central

    Feld, Geoffrey K; Brown, Michael J; Krantz, Bryan A

    2012-01-01

    Energy-consuming nanomachines catalyze the directed movement of biopolymers in the cell. They are found both dissolved in the aqueous cytosol as well as embedded in lipid bilayers. Inquiries into the molecular mechanism of nanomachine-catalyzed biopolymer transport have revealed that these machines are equipped with molecular parts, including adjustable clamps, levers, and adaptors, which interact favorably with substrate polypeptides. Biological nanomachines that catalyze protein transport, known as translocases, often require that their substrate proteins unfold before translocation. An unstructured protein chain is likely entropically challenging to bind, push, or pull in a directional manner, especially in a way that produces an unfolding force. A number of ingenious solutions to this problem are now evident in the anthrax toxin system, a model used to study protein translocation. Here we highlight molecular ratchets and current research on anthrax toxin translocation. A picture is emerging of proton-gradient-driven anthrax toxin translocation, and its associated ratchet mechanism likely applies broadly to other systems. We suggest a cyclical thermodynamic order-to-disorder mechanism (akin to a heat-engine cycle) is central to underlying protein translocation: peptide substrates nonspecifically bind to molecular clamps, which possess adjustable affinities; polypeptide substrates compress into helical structures; these clamps undergo proton-gated switching; and the substrate subsequently expands regaining its unfolded state conformational entropy upon translocation. PMID:22374876

  4. The assay of diphtheria toxin

    PubMed Central

    Gerwing, Julia; Long, D. A.; Mussett, Marjorie V.

    1957-01-01

    A precise assay of diphtheria toxin is described, based on the linear relationship between the diameter of the skin reaction to, and logarithm of the dose of, toxin. It eliminates the need for preliminary titrations, is economical, provides information about the slope of the log-dose response lines and, therefore, of the validity of the assay, and yields limits of error of potency from the internal evidence of the assay. A study has been made of the effects of avidity, combining power, toxicity and buffering on the assay of diphtheria toxins against the International Standards for both Diphtheria Antitoxin and Schick-Test Toxin. All the toxins assayed against the standard toxin, whatever their other properties might be, gave log-dose response lines of similar slope provided that they were diluted in buffered physiological saline. The assays were therefore valid. These experiments were repeated concurrently in non-immune and in actively immunized guinea-pigs, and comparable figures for potency obtained in both groups. The result was not significantly affected by the avidity or combining power of the toxin. However, non-avid toxins gave low values in Schick units when assayed, by the Rmer & Sames technique, in terms of the International Standard for Diphtheria Antitoxin. The problem of the ultimate standard and the implications of these findings are discussed. PMID:13511133

  5. Botulinum toxin: Bioweapon & magic drug

    PubMed Central

    Dhaked, Ram Kumar; Singh, Manglesh Kumar; Singh, Padma; Gupta, Pallavi

    2010-01-01

    Botulinum neurotoxins, causative agents of botulism in humans, are produced by Clostridium botulinum, an anaerobic spore-former Gram positive bacillus. Botulinum neurotoxin poses a major bioweapon threat because of its extreme potency and lethality; its ease of production, transport, and misuse; and the need for prolonged intensive care among affected persons. A single gram of crystalline toxin, evenly dispersed and inhaled, can kill more than one million people. The basis of the phenomenal potency of botulinum toxin is enzymatic; the toxin is a zinc proteinase that cleaves neuronal vesicle associated proteins responsible for acetylcholine release into the neuromuscular junction. As a military or terrorist weapon, botulinum toxin could be disseminated via aerosol or by contamination of water or food supplies, causing widespread casualties. A fascinating aspect of botulinum toxin research in recent years has been development of the most potent toxin into a molecule of significant therapeutic utility. It is the first biological toxin which is licensed for treatment of human diseases. In the late 1980s, Canada approved use of the toxin to treat strabismus, in 2001 in the removal of facial wrinkles and in 2002, the FDA in the United States followed suit. The present review focuses on both warfare potential and medical uses of botulinum neurotoxin. PMID:21149997

  6. TOXINS FROM CYANOBACTERIA IN WATER

    EPA Science Inventory

    This project is part of a larger U. S. Environmental Protection Agency (EPA) effort, which includes the Office of Water, to investigate algal toxins in surface water supplies and drinking water. Toxins produced by cyanobacteria (blue-green algae) are among the most potent known ...

  7. Lymphocyte receptors for pertussis toxin

    SciTech Connect

    Clark, C.G.; Armstrong, G.D. )

    1990-12-01

    We have investigated human T-lymphocyte receptors for pertussis toxin by affinity isolation and photoaffinity labeling procedures. T lymphocytes were obtained from peripheral human blood, surface iodinated, and solubilized in Triton X-100. The iodinated mixture was then passed through pertussis toxin-agarose, and the fractions were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Autoradiography of the fixed, dried gels revealed several bands in the pertussis toxin-bound fraction that were not observed in fractions obtained from histone or fetuin-agarose. Further investigations employed a photoaffinity labeling reagent, sulfosuccinimidyl 2-(p-azido-salicylamido)-1,3'-dithiopropionate, to identify pertussis toxin receptors in freshly isolated peripheral blood monocytic cells, T lymphocytes, and Jurkat cells. In all three cell systems, the pertussis toxin affinity probe specifically labeled a single protein species with an apparent molecular weight of 70,000 that was not observed when the procedure was performed in the presence of excess unmodified pertussis toxin. A protein comparable in molecular weight to the one detected by the photoaffinity labeling technique was also observed among the species that bound to pertussis toxin-agarose. The results suggest that pertussis toxin may bind to a 70,000-Da receptor in human T lymphocytes.

  8. B-cell epitope of beta toxin of Clostridium perfringens genetically conjugated to a carrier protein: expression, purification and characterization of the chimeric protein.

    PubMed

    Bhatia, Bharti; Solanki, Amit Kumar; Kaushik, Himani; Dixit, Aparna; Garg, Lalit C

    2014-10-01

    Beta toxin (btx) is the prime virulence factor for the pathogenesis of Clostridium perfringens type C strain, known to cause necrotic enteritis and enterotoxaemia in mammalian species. The existing vaccines targeting btx are formaldehyde inactivated culture filtrates of Clostridium. These filtrates raise antigenic load in the host leading to nonspecific and poor responses. The present study aimed to overcome these drawbacks and generate a chimeric protein carrying in silico identified B-cell epitope of btx fused with a carrier protein as a vaccine candidate. Using bioinformatic tools, three stretches of amino acids were predicted as putative B-cell epitopes. One of the epitopes spanning 140-156 amino acid residues was genetically conjugated with B-subunit of heat labile enterotoxin (LTB) of Escherichia coli and expressed as a translational fusion in Vibrio cholerae secretory expression system. High level expression of the recombinant fusion protein rLTB-Btx140-156 was obtained and the protein was successfully purified. The recombinant protein retained the native LTB property to pentamerize and bind to GM1 ganglioside receptor of LTB. The antigenicity of both the epitope and the carrier protein was maintained in fusion protein as indicated by immunoblotting against anti-LTB and anti-btx antibody. The rLTB-Btx140-156 fusion protein therefore can be evaluated as a potential vaccine candidate against C. perfringens. PMID:24996028

  9. Toxin production by Campylobacter spp.

    PubMed Central

    Wassenaar, T M

    1997-01-01

    Of all the virulence factors that were proposed for Campylobacter jejuni and related species to cause disease in humans, the discovery of toxin production was the most promising but led to a rather confusing and even disappointing stream of data. The discussion of whether proteinaceous exotoxins are relevant in disease remains open. One important reason for this lack of consensus is the anecdotal nature of the literature reports. To provide a basis for an unbiased opinion, this review compiles all described exotoxins, compares their reported properties, and provides a summary of animal model studies and clinical data. The toxins are divided into enterotoxins and cytotoxins and are sorted according to their biochemical properties. Since many Campylobacter toxins have been compared with toxins of other species, some key examples of the latter are also discussed. Future directions of toxin research that appear promising are defined. PMID:9227862

  10. Toxin-Based Therapeutic Approaches

    PubMed Central

    Shapira, Assaf; Benhar, Itai

    2010-01-01

    Protein toxins confer a defense against predation/grazing or a superior pathogenic competence upon the producing organism. Such toxins have been perfected through evolution in poisonous animals/plants and pathogenic bacteria. Over the past five decades, a lot of effort has been invested in studying their mechanism of action, the way they contribute to pathogenicity and in the development of antidotes that neutralize their action. In parallel, many research groups turned to explore the pharmaceutical potential of such toxins when they are used to efficiently impair essential cellular processes and/or damage the integrity of their target cells. The following review summarizes major advances in the field of toxin based therapeutics and offers a comprehensive description of the mode of action of each applied toxin. PMID:22069564

  11. Toxin-induced hepatic injury.

    PubMed

    Lopez, Annette M; Hendrickson, Robert G

    2014-02-01

    Toxins such as pharmaceuticals, herbals, foods, and supplements may lead to hepatic damage. This damage may range from nonspecific symptoms in the setting of liver test abnormalities to acute hepatic failure. The majority of severe cases of toxin-induced hepatic injury are caused by acetaminophen and ethanol. The most important step in the patient evaluation is to gather an extensive history that includes toxin exposure and exclude common causes of liver dysfunction. Patients whose hepatic dysfunction progresses to acute liver failure may benefit from transfer to a transplant service for further management. Currently, the mainstay in management for most exposures is discontinuing the offending agent. This manuscript will review the incidence, pathophysiology, diagnosis and management of the different forms of toxin-induced hepatic injury and exam in-depth the most common hepatic toxins. PMID:24275171

  12. Binding of ATP by pertussis toxin and isolated toxin subunits

    SciTech Connect

    Hausman, S.Z.; Manclark, C.R.; Burns, D.L. )

    1990-07-03

    The binding of ATP to pertussis toxin and its components, the A subunit and B oligomer, was investigated. Whereas, radiolabeled ATP bound to the B oligomer and pertussis toxin, no binding to the A subunit was observed. The binding of ({sup 3}H)ATP to pertussis toxin and the B oligomer was inhibited by nucleotides. The relative effectiveness of the nucleotides was shown to be ATP > GTP > CTP > TTP for pertussis toxin and ATP > GTP > TTP > CTP for the B oligomer. Phosphate ions inhibited the binding of ({sup 3}H)ATP to pertussis toxin in a competitive manner; however, the presence of phosphate ions was essential for binding of ATP to the B oligomer. The toxin substrate, NAD, did not affect the binding of ({sup 3}H)ATP to pertussis toxin, although the glycoprotein fetuin significantly decreased binding. These results suggest that the binding site for ATP is located on the B oligomer and is distinct from the enzymatically active site but may be located near the eukaryotic receptor binding site.

  13. Comparison of Outcomes in Patients With Nonobstructive, Labile-Obstructive, and Chronically Obstructive Hypertrophic Cardiomyopathy.

    PubMed

    Pozios, Iraklis; Corona-Villalobos, Celia; Sorensen, Lars L; Bravo, Paco E; Canepa, Marco; Pisanello, Chiara; Pinheiro, Aurelio; Dimaano, Veronica L; Luo, Hongchang; Dardari, Zeina; Zhou, Xun; Kamel, Ihab; Zimmerman, Stefan L; Bluemke, David A; Abraham, M Roselle; Abraham, Theodore P

    2015-09-15

    Patients with nonobstructive hypertrophic cardiomyopathy (HC) are considered low risk, generally not requiring aggressive intervention. However, nonobstructive and labile-obstructive HC have been traditionally classified together, and it is unknown if these 2 subgroups have distinct risk profiles. We compared cardiovascular outcomes in 293 patients HC (96 nonobstructive, 114 labile-obstructive, and 83 obstructive) referred for exercise echocardiography and magnetic resonance imaging and followed for 3.3 3.6 years. A subgroup (34 nonobstructive, 28 labile-obstructive, 21 obstructive) underwent positron emission tomography. The mean number of sudden cardiac death risk factors was similar among groups (nonobstructive: 1.4 vs labile-obstructive: 1.2 vs obstructive: 1.4 risk factors, p = 0.2). Prevalence of late gadolinium enhancement (LGE) was similar across groups but more non-obstructive patients had late gadolinium enhancement ?20% of myocardial mass (23 [30%] vs 19 [18%] labile-obstructive and 8 [11%] obstructive, p = 0.01]. Fewer labile-obstructive patients had regional positron emission tomography perfusion abnormalities (12 [46%] vs nonobstructive 30 [81%] and obstructive 17 [85%], p = 0.003]. During follow-up, 60 events were recorded (36 ventricular tachycardia/ventricular fibrillation, including 30 defibrillator discharges, 12 heart failure worsening, and 2 deaths). Nonobstructive patients were at greater risk of VT/VF at follow-up, compared to labile obstructive (hazed ratio 0.18, 95% confidence interval 0.04 to 0.84, p = 0.03) and the risk persisted after adjusting for age, gender, syncope, family history of sudden cardiac death, abnormal blood pressure response, and septum ?3 cm (p = 0.04). Appropriate defibrillator discharges were more frequent in nonobstructive (8 [18%]) compared to labile-obstructive (0 [0%], p = 0.02) patients. In conclusion, nonobstructive hemodynamics is associated with more pronounced fibrosis and ischemia than labile-obstructive and is an independent predictor of VT/VF in HC. PMID:26239580

  14. Continuous flow analysis of labile iron in ice-cores.

    PubMed

    Hiscock, William T; Fischer, Hubertus; Bigler, Matthias; Gfeller, Gideon; Leuenberger, Daiana; Mini, Olivia

    2013-05-01

    The important active and passive role of mineral dust aerosol in the climate and the global carbon cycle over the last glacial/interglacial cycles has been recognized. However, little data on the most important aeolian dust-derived biological micronutrient, iron (Fe), has so far been available from ice-cores from Greenland or Antarctica. Furthermore, Fe deposition reconstructions derived from the palaeoproxies particulate dust and calcium differ significantly from the Fe flux data available. The ability to measure high temporal resolution Fe data in polar ice-cores is crucial for the study of the timing and magnitude of relationships between geochemical events and biological responses in the open ocean. This work adapts an existing flow injection analysis (FIA) methodology for low-level trace Fe determinations with an existing glaciochemical analysis system, continuous flow analysis (CFA) of ice-cores. Fe-induced oxidation of N,N'-dimethyl-p-pheylenediamine (DPD) is used to quantify the biologically more important and easily leachable Fe fraction released in a controlled digestion step at pH ~1.0. The developed method was successfully applied to the determination of labile Fe in ice-core samples collected from the Antarctic Byrd ice-core and the Greenland Ice-Core Project (GRIP) ice-core. PMID:23594184

  15. The labile iron pool in human erythroid cells.

    PubMed

    Prus, Eugenia; Fibach, Eitan

    2008-06-01

    Although most cellular iron is firmly bound (e.g. in haemoglobin), some, the labile iron pool (LIP), is bound to low-affinity ligands. The LIP is regarded as the crossroads of cellular iron traffic. Using multi-parameter flow-cytometry of cells treated with the metal-sensitive sensor calcein and the cell-permeable chelator deferiprone, we studied LIP in various human erythroid cell populations in peripheral blood, bone marrow and culture. Erythroid maturation was found to be associated with a decrease in the LIP. In the peripheral blood, nucleated erythrocytes (normoblasts) had 5.8-fold and 8.8-fold greater LIP than reticulocytes and erythrocytes respectively. Early reticulocytes had 2.5-fold more LIP than late reticulocytes. In the bone-marrow and in culture, LIP decreased by c. 30-fold as early erythroid precursors matured to late precursors. Adding holo-transferrin to iron-depleted cultures elevated LIP by 3.9-fold. We also show that in beta-thalassaemia, a disease associated with iron-overload, erythrocytes and reticulocytes in the blood and erythroid precursors in culture have a significantly greater LIP than their normal counterparts. In conclusion, the LIP in erythroid cells is altered under physiological (maturation) and pathological (thalassaemia) conditions. The methodology presented might be useful for evaluating the LIP in various diseases and for studying the efficacy of iron-chelators. PMID:18503586

  16. High Sensitivity Combined with Extended Structural Coverage of Labile Compounds via Nanoelectrospray Ionization at Subambient Pressures

    SciTech Connect

    Cox, Jonathan T.; Kronewitter, Scott R.; Shukla, Anil K.; Moore, Ronald J.; Smith, Richard D.; Tang, Keqi

    2014-10-07

    Subambient pressure ionization with nanoelectrospray (SPIN) has proven to be effective in producing ions with high efficiency and transmitting them to low pressures for high sensitivity mass spectrometry (MS) analysis. Here we present evidence that not only does the SPIN source improve MS sensitivity but also allows for gentler ionization conditions. The gentleness of a conventional heated capillary electrospray ionization (ESI) source and the SPIN source was compared by the liquid chromatography mass spectrometry (LC-MS) analysis of colominic acid. Colominic acid is a mixture of sialic acid polymers of different lengths containing labile glycosidic linkages between monomer units necessitating a gentle ion source. By coupling the SPIN source with high resolution mass spectrometry and using advanced data processing tools, we demonstrate much extended coverage of sialic acid polymer chains as compared to using the conventional ESI source. Additionally we show that SPIN-LC-MS is effective in elucidating polymer features with high efficiency and high sensitivity previously unattainable by the conventional ESI-LC-MS methods.  

  17. Alternaria brassicae produces a host-specific protein toxin from germinating spores on host leaves.

    PubMed

    Parada, R Y; Sakuno, E; Mori, N; Oka, K; Egusa, M; Kodama, M; Otani, H

    2008-04-01

    Spore suspensions of Alternaria brassicae, the causal agent of gray leaf spot in Brassica plants, were incubated on the leaves of cabbage (B. oleracea) and spore germination fluid (SGF) was collected after 48 h. A high molecular weight (HMW) fraction (>10 kDa) was separated from the SGF by ultrafiltration. In a detached leaf assay, the HMW fraction induced visible symptoms only on host leaves and the toxicity was lost by treatment with proteinase K or heat at 60 degrees C for 15 min, indicating the presence of host-specific protein toxin(s). A protein toxin in the HMW fraction was purified by several chromatography steps. The toxin induced water-soaked symptoms followed by chlorosis at concentrations of 0.5 to 1 microg/ml on host leaves, but not on nonhost leaves even at 50 microg/ml. The toxin also had infection-inducing activity when added to spore suspension of a nonpathogenic isolate of A. alternata, causing symptoms similar to the infection of A. brassicae only on host leaves. These results indicate that a new host-specific protein toxin named ABR-toxin is released from germinating spores of A. brassicae on host leaves. ABR-toxin migrated as a protein of 27.5 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The isoelectric point of ABR-toxin was estimated to be approximately 7.0 and 21 N-terminal amino acid residues were sequenced. PMID:18944195

  18. Stoichiometric regulation of phytoplankton toxins.

    PubMed

    Van de Waal, Dedmer B; Smith, Val H; Declerck, Steven A J; Stam, Eva C M; Elser, James J

    2014-06-01

    Ecological Stoichiometry theory predicts that the production, elemental structure and cellular content of biomolecules should depend on the relative availability of resources and the elemental composition of their producer organism. We review the extent to which carbon- and nitrogen-rich phytoplankton toxins are regulated by nutrient limitation and cellular stoichiometry. Consistent with theory, we show that nitrogen limitation causes a reduction in the cellular quota of nitrogen-rich toxins, while phosphorus limitation causes an increase in the most nitrogen-rich paralytic shellfish poisoning toxin. In addition, we show that the cellular content of nitrogen-rich toxins increases with increasing cellular N : P ratios. Also consistent with theory, limitation by either nitrogen or phosphorus promotes the C-rich toxin cell quota or toxicity of phytoplankton cells. These observed relationships may assist in predicting and managing toxin-producing phytoplankton blooms. Such a stoichiometric regulation of toxins is likely not restricted to phytoplankton, and may well apply to carbon- and nitrogen-rich secondary metabolites produced by bacteria, fungi and plants. PMID:24712512

  19. Stoichiometric regulation of phytoplankton toxins.

    TOXLINE Toxicology Bibliographic Information

    Van de Waal DB; Smith VH; Declerck SA; Stam EC; Elser JJ

    2014-06-01

    Ecological Stoichiometry theory predicts that the production, elemental structure and cellular content of biomolecules should depend on the relative availability of resources and the elemental composition of their producer organism. We review the extent to which carbon- and nitrogen-rich phytoplankton toxins are regulated by nutrient limitation and cellular stoichiometry. Consistent with theory, we show that nitrogen limitation causes a reduction in the cellular quota of nitrogen-rich toxins, while phosphorus limitation causes an increase in the most nitrogen-rich paralytic shellfish poisoning toxin. In addition, we show that the cellular content of nitrogen-rich toxins increases with increasing cellular N : P ratios. Also consistent with theory, limitation by either nitrogen or phosphorus promotes the C-rich toxin cell quota or toxicity of phytoplankton cells. These observed relationships may assist in predicting and managing toxin-producing phytoplankton blooms. Such a stoichiometric regulation of toxins is likely not restricted to phytoplankton, and may well apply to carbon- and nitrogen-rich secondary metabolites produced by bacteria, fungi and plants.

  20. Food toxin detection with atomic force microscope

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Externally introduced toxins or internal spoilage correlated pathogens and their metabolites are all potential sources of food toxins. To prevent and protect unsafe food, many food toxin detection techniques have been developed to detect various toxins for quality control. Although several routine m...

  1. [Shiga toxin and tetanus toxin as a potential biologic weapon].

    PubMed

    Toczyska, Izabela; P?usa, Tadeusz

    2015-09-01

    Toxins produced by the bacteria are of particular interest as potential cargo combat possible for use in a terrorist attack or war. Shiga toxin is usually produced by shiga toxigenic strains of Escherichia coli (STEC - shigatoxigenic Escherichia coli). To infection occurs mostly after eating contaminated beef. Clinical syndromes associated with Shiga toxin diarrhea, hemorrhagic colitis, hemolytic uremic syndrome (HUS - hemolytic uremic syndrome) or thrombotic thrombocytopenic purpura. Treatment is symptomatic. In HUS, in which mortality during an epidemic reaches 20%, extending the kidney injury dialysis may be necessary. Exposure to tetanus toxin produced by Clostridium tetani, resulting in the most generalized tetanus, characterized by increased muscle tension and painful contractions of individual muscle groups. In the treatment beyond symptomatic behavior (among others spasticity medications, anticonvulsants, muscle relaxants) is used tetanus antitoxin and antibiotics (metronidazole choice). A common complication is acute respiratory failure - then it is necessary to implement mechanical ventilation. PMID:26449578

  2. The three-dimensional crystal structure of cholera toxin

    SciTech Connect

    Zhang, Rong-Guang; Westbrook, M.L.; Nance, S.; Spangler, B.D.; Scott, D.L.; Westbrook, E.M.

    1996-02-01

    The clinical manifestations of cholera are largely attributable to the actions of a secreted hexameric AB{sub 5} enterotoxin (choleragen). We have solved the three-dimensional structure of choleragen at 2.5 {Angstrom} resolution and compared the refined coordinates with those of choleragenoid (isolated B pentamer) and the heat-labile enterotoxin from Escherichia coli (LT). The crystalline coordinates provide a detailed view of the stereochemistry implicated in binding to GM1 gangliosides and in carrying out ADP-ribosylation. The A2 chain of choleragen, in contrast to that of LT, is a nearly continuous {alpha}-helix with an interpretable carboxyl tail.

  3. DETERMINATION OF APPARENT QUANTUM YIELD SPECTRA FOR THE FORMATION OF BIOLOGICALLY LABILE PHOTOPRODUCTS

    EPA Science Inventory

    Quantum yield spectra for the photochemical formation of biologically labile photoproducts from dissolved organic matter (DOM) have not been available previously, although they would greatly facilitate attempts to model photoproduct formation rates across latitudinal, seasonal, a...

  4. Biogeochemical implications of labile phosphorus in forest soils determined by the Hedley fractionation procedure.

    PubMed

    Johnson, Arthur H; Frizano, Jaqueline; Vann, David R

    2003-05-01

    Forest ecologists and biogeochemists have used a variety of extraction techniques to assess labile vs. non-labile soil P pools in chronosequences, the balance between biological vs. geochemical control of P transformations across a wide range of soil orders, the role of plants with either N-fixing or mycorrhizal symbionts in controlling soil P fractions, and to make inferences about plant-available P. Currently, variants of the sequential extraction procedure developed by M. J. Hedley and co-workers afford the greatest discrimination among labile and non-labile organic and inorganic P pools. Results of recent studies that used this technique to evaluate P fractions in forest soils indicate the following: (1) in intact, highly weathered forest soils of the humid tropics, Hedley-labile P values are several times larger than extractable P values resulting from mildly acidic extracting solutions which were commonly used in the past 2 decades; (2) pools of Hedley-labile P are several times larger than the annual forest P requirement and P required from the soil annually in both temperate and tropical forests; (3) long-term trends in non-labile P pools during pedogenesis are adequately represented by the Walker and Syers' model of changes in P fractionation during soil development. However, to better represent trends in pools that can supply plant-available P across forest soils of different age and weathering status, the paradigm should be modified; and (4) across a wide range of tropical and temperate forest soils, organic matter content is an important determinant of Hedley-labile P. PMID:12695899

  5. A general approach to anionic acid-labile surfactants with tunable properties.

    PubMed

    Li, Miaosheng; Powell, Matthew J; Razunguzwa, Trust T; O'Doherty, George A

    2010-09-17

    A general approach to the synthesis of a new series of unique sulfate anionic acid-labile surfactants (AALS) was developed. In this approach, the ketal was derived from methyl pyruvate, and the sulfate motif was introduced via sulfitylation of the alcohol, oxidation, and finally conversion of the sulfate diester to the desired sodium salt. The physicochemical properties in aqueous solution of this novel series of surfactants, such as CMCs, solubility, acid lability, and stability were studied. PMID:20726615

  6. Thermally-Labile Trace Elements in Enstatite Meteorites

    NASA Technical Reports Server (NTRS)

    Wang, M.-S.; Lipschutz, M. E.

    2000-01-01

    RNAA data for Bi, In and Tl in 30 E3-6 chondrites accord well with trends for heated Abee (EH4) suggesting that all EH and EL samples reflect open-system, post-accretionary heating, independent of siderophile content or recovery location.

  7. Controls on diurnal variation in labile soil phosphorus of a humid tropical forest, Puerto Rico

    NASA Astrophysics Data System (ADS)

    Wood, T. E.; Vandecar, K. L.; Matthews, D.; Lawrence, D.

    2011-12-01

    Tropical forests are typically considered phosphorous (P) limited. Variability in nutrient availability has been shown to affect forest productivity and foliar nutrient concentrations, as well as the rate of soil carbon (C) loss via effects on decomposition and soil respiration rates. It follows that the C efflux and long-term stability of C stocks in tropical soils is likely to depend on nutrient availability. We quantified short-term variability (hours to days) in labile P in a Puerto Rican wet tropical forest and investigated potential environmental and biotic controls such as soil moisture, soil temperature, light and soil respiration. We measured hourly changes in labile P from sunrise to sunset on five separate days in July of 2008. Labile P varied significantly both within and among days. Concentrations of labile P ranged from 2.75-3.75 ?g/g. Labile P was positively related to volumetric soil moisture (R2= 0.3, p<0.05). These results suggest that the labile P pool is highly dynamic on short timescales, even in relatively a-seasonal tropical forest.

  8. Maternal emotion socialization differentially predicts third-grade children's emotion regulation and lability.

    PubMed

    Rogers, Megan L; Halberstadt, Amy G; Castro, Vanessa L; MacCormack, Jennifer K; Garrett-Peters, Patricia

    2016-03-01

    Numerous parental emotion socialization factors have been implicated as direct and indirect contributors to the development of children's emotional competence. To date, however, no study has combined parents' emotion-related beliefs, behaviors, and regulation strategies in one model to assess their cumulative-as well as unique-contributions to children's emotion regulation. We considered the 2 components that have recently been distinguished: emotion regulation and emotional lability. We predicted that mothers' beliefs about the value of and contempt for children's emotions, mothers' supportive and nonsupportive reactions to their children's emotions, as well as mothers' use of cognitive reappraisal and suppression of their own emotions would each contribute unique variance to their children's emotion regulation and lability, as assessed by children's teachers. The study sample consisted of an ethnically and socioeconomically diverse group of 165 mothers and their third-grade children. Different patterns emerged for regulation and lability: Controlling for family income, child gender, and ethnicity, only mothers' lack of suppression as a regulatory strategy predicted greater emotion regulation in children, whereas mothers' valuing of children's emotions, mothers' lack of contempt for children's emotions, mothers' use of cognitive reappraisal to reinterpret events, and mothers' lack of emotional suppression predicted less lability in children. These findings support the divergence of emotion regulation and lability as constructs and indicate that, during middle childhood, children's lability may be substantially and uniquely affected by multiple forms of parental socialization. (PsycINFO Database Record PMID:26641269

  9. Analyzing a bioterror attack on the food supply: The case of botulinum toxin in milk

    PubMed Central

    Wein, Lawrence M.; Liu, Yifan

    2005-01-01

    We developed a mathematical model of a cows-to-consumers supply chain associated with a single milk-processing facility that is the victim of a deliberate release of botulinum toxin. Because centralized storage and processing lead to substantial dilution of the toxin, a minimum amount of toxin is required for the release to do damage. Irreducible uncertainties regarding the doseresponse curve prevent us from quantifying the minimum effective release. However, if terrorists can obtain enough toxin, and this may well be possible, then rapid distribution and consumption result in several hundred thousand poisoned individuals if detection from early symptomatics is not timely. Timely and specific in-process testing has the potential to eliminate the threat of this scenario at a cost of <1 cent per gallon and should be pursued aggressively. Investigation of improving the toxin inactivation rate of heat pasteurization without sacrificing taste or nutrition is warranted. PMID:15985558

  10. A Bivalent Tarantula Toxin Activates the Capsaicin Receptor, TRPV1, by Targeting the Outer Pore Domain

    PubMed Central

    Bohlen, Christopher J.; Priel, Avi; Zhou, Sharleen; King, David; Siemens, Jan; Julius, David

    2010-01-01

    SUMMARY Toxins have evolved to target regions of membrane ion channels that underlie ligand binding, gating, or ion permeation, and have thus served as invaluable tools for probing channel structure and function. Here we describe a peptide toxin from the Earth Tiger tarantula that selectively and irreversibly activates the capsaicin- and heat-sensitive channel, TRPV1. This high avidity interaction derives from a unique tandem repeat structure of the toxin that endows it with an antibody-like bivalency, illustrating a new paradigm in toxin structure and evolution. The double-knot toxin traps TRPV1 in the open state by interacting with residues in the presumptive pore-forming region of the channel, highlighting the importance of conformational changes in the outer pore region of TRP channels during activation. PMID:20510930

  11. Isolation of B subunit-specific monoclonal antibody clones that strongly neutralize the toxicity of Shiga toxin 2.

    PubMed

    Arimitsu, Hideyuki; Sasaki, Keiko; Iba, Yoshitaka; Kurosawa, Yoshikazu; Shimizu, Toshiyasu; Tsuji, Takao

    2015-02-01

    Shiga toxin 2 (Stx2)-specific mAb-producing hybridoma clones were generated from mice. Because mice tend to produce small amounts of B subunit (Stx2B)-specific antibodies at the polyclonal antibody level after immunization via the parenteral route, mice were immunized intranasally with Stx2 toxoids with a mutant heat-labile enterotoxin as a mucosal adjuvant; 11 different hybridoma clones were obtained in two trials. Six of them were A subunit (Stx2A)-specific whereas five were Stx2B-specific antibody-producing clones. The in vitro neutralization activity of Stx2B-specific mAbs against Stx2 was greater than that of Stx2A-specific mAbs on HeLa229 cells. Furthermore, even at low concentrations two of the Stx2B-specific mAbs (45 and 75D9) completely inhibited receptor binding and showed in vivo neutralization activity against a fivefold median lethal dose of Stx2 in mice. In western blot analysis, these Stx2B-specific neutralization antibodies did not react to three different mutant forms of Stx2, each amino acid residue of which was associated with receptor binding. Additionally, the nucleotide sequences of the VH and VL regions of clones 45 and 75D9 were determined. Our Stx2B-specific mAbs may be new candidates for the development of mouse-human chimeric Stx2-neutralizing antibodies which have fewer adverse effects than animal antibodies for enterohemorrhagic Escherichia coli infection. PMID:25521016

  12. [Today's threat of ricin toxin].

    PubMed

    From, Sławomir; Płusa, Tadeusz

    2015-09-01

    Since the late 70s of the last century there were more than 700 incidents related to the use of the ricin toxin. For this reason, CDC (Center of Disease Control and Prevention) recognized toxin as a biological weapon category B. The lethal dose of ricin toxin after parenteral administration is 0.0001 mg/kg and after oral administration 0.2 mg. The first symptoms of poisoning occur within a few hours after application of toxin as a nausea, vomiting and abdominal pain. In the final stage there are observed: cardiac arrhythmia, collapse and symptoms suggestive of involvement of the central nervous system. Stage immediately preceding death is a state of coma. The ricin toxin is still the substance against which action has no optimal antidote. Developed a vaccine called RiVax is waiting for its registration. It should be pointed out that the availability of a ricin toxin makes it possible to use it for real bioterrorists. PMID:26449579

  13. Thermally labile components of aqueous humor potently induce osteogenic potential in adipose-derived mesenchymal stem cells.

    PubMed

    Morgan, Joshua T; Kwon, Heung Sun; Wood, Joshua A; Borjesson, Dori L; Tomarev, Stanislav I; Murphy, Christopher J; Russell, Paul

    2015-06-01

    Adipose-derived mesenchymal stem cells (ASCs) hold promise for use in cell-based therapies. Their intrinsic anti-inflammatory properties are potentially useful for treatments of inflammatory conditions such as uveitis, while their ability to differentiate along multiple cell lineages suggests use in regenerating damaged or degenerated tissue. However, how ASCs will respond to the intraocular environment is poorly studied. We have recently reported that aqueous humor (AH), the fluid that nourishes the anterior segment of the eye, potently increases alkaline phosphatase (ALP) activity of ASCs, indicating osteogenic differentiation. Here, we expand on our previous findings to better define the nature of this response. To this end, we cultured ASCs in the presence of 0, 5, 10, and 20% AH and assayed them for ALP activity. We found ALP activity correlates with increasing AH concentrations from 5 to 20%, and that longer treatments result in increased ALP activity. By using serum free media and pretreating AH with dextran-coated charcoal, we found that serum and charcoal-adsorbable AH components augment but are not required for this response. Further, by heat-treating the AH, we established that thermally labile components are required for the osteogenic response. Finally, we showed myocilin, a protein present in AH, could induce ALP activity in ASCs. However, this was to a lesser extent than untreated 5% AH, and myocilin could only partially rescue the effect after heat treatment, documenting there were additional thermally labile constituents of AH involved in the osteogenic response. Our work adds to the understanding of the induction of ALP in ASCs following exposure to AH, providing important insight in how ASCs will be influenced by the ocular environment. In conclusion, increased osteogenic potential upon exposure to AH represents a potential challenge to developing ASC cell-based therapies directed at the eye. PMID:25720657

  14. Induction of apoptosis by Shiga toxins

    PubMed Central

    Tesh, Vernon L

    2010-01-01

    Shiga toxins comprise a family of structurally and functionally related protein toxins expressed by Shigella dysenteriae serotype 1 and multiple serotypes of Escherichia coli. While the capacity of Shiga toxins to inhibit protein synthesis by catalytic inactivation of eukaryotic ribosomes has been well described, it is also apparent that Shiga toxins trigger apoptosis in many cell types. This review presents evidence that Shiga toxins induce apoptosis of epithelial, endothelial, leukocytic, lymphoid and neuronal cells. Apoptotic signaling pathways activated by the toxins are reviewed with an emphasis on signaling mechanisms that are shared among different cell types. Data suggesting that Shiga toxins induce apoptosis through the endoplasmic reticulum stress response and clinical evidence demonstrating apoptosis in humans infected with Shiga toxin-producing bacteria are briefly discussed. The potential for use of Shiga toxins to induce apoptosis in cancer cells is briefly reviewed. PMID:20210553

  15. Antibody-based biological toxin detection

    SciTech Connect

    Menking, D.E.; Goode, M.T.

    1995-12-01

    Fiber optic evanescent fluorosensors are under investigation in our laboratory for the study of drug-receptor interactions for detection of threat agents and antibody-antigen interactions for detection of biological toxins. In a direct competition assay, antibodies against Cholera toxin, Staphylococcus Enterotoxin B or ricin were noncovalently immobilized on quartz fibers and probed with fluorescein isothiocyanate (FITC) - labeled toxins. In the indirect competition assay, Cholera toxin or Botulinum toxoid A was immobilized onto the fiber, followed by incubation in an antiserum or partially purified anti-toxin IgG. These were then probed with FITC-anti-IgG antibodies. Unlabeled toxins competed with labeled toxins or anti-toxin IgG in a dose dependent manner and the detection of the toxins was in the nanomolar range.

  16. Influence of yogurt fermentation and refrigerated storage on the stability of protein toxin contaminants.

    PubMed

    Jackson, Lauren S; Triplett, Odbert A; Tolleson, William H

    2015-06-01

    Dairy products sold in a ready-to-eat form present the risk that adulterants persisting through manufacturing, storage, and distribution would reach consumers. Pathogenic microbes, including shigatoxigenic strains of Escherichia coli and the toxins they produce, are common food safety hazards associated with dairy products. Ricin and abrin are plant-derived ribosome-inactivating protein toxins related to the shiga-like toxins produced by E. coli. Limited information exists on the effects of manufacturing processes on the stabilities of these heat-resistant ribosome-inactivating proteins in the presence of foods. The goal of this study was to determine how typical yogurt manufacturing and storage processes influence ribosome-inactivating protein toxins. Ricin and abrin were added to skim or whole milk and batch pasteurized. Complete inactivation of both toxins was observed after 30 minutes at 85 °C. If the toxins were added after pasteurization, the levels of ricin and abrin in yogurt and their cytotoxic activities did not change significantly during fermentation or refrigerated storage for 4 weeks. The activities of ricin and abrin were inhibited by skim milk, nonfat yogurt, whole milk, and whole milk yogurt. The results showed minimal effects of the toxins on yogurt pH and %titratable acidity but inhibitory effects of yogurt on toxin activity. PMID:25772284

  17. Scorpion toxins prefer salt solutions.

    PubMed

    Nikouee, Azadeh; Khabiri, Morteza; Cwiklik, Lukasz

    2015-11-01

    There is a wide variety of ion channel types with various types of blockers, making research in this field very complicated. To reduce this complexity, it is essential to study ion channels and their blockers independently. Scorpion toxins, a major class of blockers, are charged short peptides with high affinities for potassium channels. Their high selectivity and inhibitory properties make them an important pharmacological tool for treating autoimmune or nervous system disorders. Scorpion toxins typically have highly charged surfaces and-like other proteins-an intrinsic ability to bind ions (Friedman J Phys Chem B 115(29):9213-9223, 1996; Baldwin Biophys J 71(4):2056-2063, 1996; Vrbka et al. Proc Natl Acad Sci USA 103(42):15440-15444, 2006a; Vrbka et al. J Phys Chem B 110(13):7036-43, 2006b). Thus, their effects on potassium channels are usually investigated in various ionic solutions. In this work, computer simulations of protein structures were performed to analyze the structural properties of the key residues (i.e., those that are presumably involved in contact with the surfaces of the ion channels) of 12 scorpion toxins. The presence of the two most physiologically abundant cations, Na(+) and K(+), was considered. The results indicated that the ion-binding properties of the toxin residues vary. Overall, all of the investigated toxins had more stable structures in ionic solutions than in water. We found that both the number and length of elements in the secondary structure varied depending on the ionic solution used (i.e., in the presence of NaCl or KCl). This study revealed that the ionic solution should be chosen carefully before performing experiments on these toxins. Similarly, the influence of these ions should be taken into consideration in the design of toxin-based pharmaceuticals. PMID:26475740

  18. Whole-Ecosystem Labile Carbon Production in a North Temperate Deciduous Forest

    NASA Astrophysics Data System (ADS)

    Gough, C. M.; Flower, C. E.; Vogel, C. S.; Dragoni, D.; Curtis, P. S.

    2008-12-01

    Management for forest carbon (C) sequestration requires knowledge of the fate of photosynthetic C. Labile C is an essential intermediary between C assimilation and growth in deciduous forests, accumulating when photosynthetic C supply exceeds demand and later depleting when reallocated to growth during periods of depressed photosynthesis. We developed a new approach that combined meteorological and biometric C cycling data for a mixed deciduous forest in Michigan, USA, to provide novel estimates of whole-ecosystem labile C production (PLC) and reallocation to growth inferred from the temporal imbalance between carbon supply from canopy net C assimilation (Ac) and C demand for net primary production (NPP). We substantiated these estimates with measurements of Populus grandidentata and Quercus rubra wood non-structural carbohydrate (NSC) concentration and mass over two years. Our analysis showed that half of annual Ac was allocated to PLC rather than to immediate growth. Labile C produced during the latter half of summer later supported dormant-season growth and respiration, with 35% of NPP in a given year requiring labile C stored during previous years. Seasonal changes in wood NSC concentration and mass generally corroborated patterns of labile C production and reallocation to growth. We observed a negative relationship between current-year PLC and NPP, indicating that disparities between same-year meteorological and biometric net ecosystem production (NEP) estimates can arise when C assimilated via photosynthesis, a flux incorporated into meteorological NEP estimates, is diverted away from NPP, a flux included in biometric NEP estimates, and instead allocated to PLC. A large, annually recharging pool of labile C also may buffer growth from climate conditions that immediately affect Ac. We conclude that a broader understanding of labile C production and reallocation across ecosystems may be important to interpreting lagged canopy C cycling and growth processes.

  19. Alteration of Labile Trace Element Concentrations in Antarctic Meteorites by Weathering: A Five-Year Assessment

    NASA Astrophysics Data System (ADS)

    Wang, M.-S.; Xiao, X.; Lipschutz, M. E.

    1992-07-01

    Numerous studies since 1987 demonstrate that, on average, Antarctic populations of specific meteorite groups differ from non-Antarctic falls. Some differences could conceivably reflect alteration during the meteorites' residence in Antarctica while others clearly are preterrestrial origin, predating fall on Earth. Concentrations of certain trace elements (Ag, Au, Bi, Cd, Co, Cs, Ga, In, Rb, Sb, Se, Te, Tl, Zn) determined by RNAA in 45 H4-6 chondrites first provided evidence for Antarctic/non-Antarctic meteorite population differences [1]. Most of these elements are thermally labile (easily lost during extended chondritic heating chondrites) so that their concentrations give important information on the thermal history of meteoritic material. Refractory elements cannot give such information. Factors possibly complicating establishment of compositional differences as preterrestrial--meteorite pairing, population reproducibility, analyst bias, and statistical modeling--are of demonstrated in consequence [1-4]. Indeed, compositional differences exist [3,4] between Antarctic meteorite populations (Victoria Land vs. Queen Maud Land) and among observed falls (Cluster 1 vs. other falls). Possibilities for meteorite compositional alteration during Antarctic weathering must be re-assessed as new data are obtained: here, we summarize the current status of this problem. 1. Highly weathered meteorites: Ten of our suite of trace elements have significantly lower mean concentrations (presumably because of leaching) in H5 chondrites of weathering types B/C and C, than in types A, A/B, and B [1]. Meteorites of types A to B-- whether exhibiting efflorescence or not--seem uncompromised [5]. 2. Antarctic meteorites of high weathering susceptibility: Carbonaceous chondrites and lunar meteorites are essentially unaltered by weathering. For example, data for LEW 90500 C(1?) chondrite reported here demonstrate that the 8 most volatile elements (Se, Cs, Te, Zn, Cd, Bi, Tl, In) have a mean Cl- normalized weight ratio of 0.585+-0.069. Other elements--even Rb, which should be easily transported in a phyllosilicate exposed to water--show no evidence for gain or loss in Antarctica. This is true also for 39 other Antarctic C2-6 chondrites [6]; 3 additional Cl-2 chondrites thermally metamorphosed in their parent bodies [7]; and lunar meteorites studied by us and others. Hydration effects are absent in these meteorites. 3. Eucrites exhibiting evidence for Ce transport: A pair of eucrite clast samples (EET 87503,23 interior and exterior), was previously studied by INAA yield REE data suggesting addition of LREE (except Ce) to the interior during Antarctic residence (Mittlefehldt, personal communication). The exterior/interior ratio for Ce, 1.1, is the same as the mean value for our RNAA trace element suite, 1.1+-0.5. Despite the large uncertainty of this ratio (reflecting the normally heterogeneous distribution of labile elements in eucrites--including falls [8]), results for EET 87503,23 are consistent with the interpretation that our suite of labile trace elements is unaffected by the process that affected REE other than Ce. Our elements are probably dispersed among many host sites, rather than being sited in a single host, like whitlockite. More RNAA measurements of additional eucrite pairs should be done to confirm this result. Further, a putative C3 clast exhibits no evidence for terrestrial alteration of RNAA elements, saponitic matrix, etc. even though REE have apparently been leached from basalts in its host eucrite, LEW 85300 [5]. After five years, numerous investigations confirm meteorite population differences consistent with the RNAA results. While Antarctic processes may have affected REE contents in some eucrites, at present no evidence exists for labile trace element transport into/out of interiors of meteorites of weathering types A to B. The absence of evidence is not evidence of absence, so continued vigilance remains necessary. Research supported by NASA grant NAG 9-48, aided by DOE grant DE-FG07-80ER10725J and NATO grant 0252/89. References: 1. Dennison J. E. and Lipschutz M. E. (1987) Geochim. Cosmochim. Acta 51, 741-754. 2. Lipschutz M. E. and Samuels S. M. (1991) Geochim. Cosmochim. Acta 55, 19-34. 3. Wolf S. F. and Lipschutz M. E. (1992) Lunar Planet. Sci. (abstract) XXIII, 1545-1546. 4. Wolf S. F. and Lipschutz M. E. (abstract), this conference. 5. Zolensky M. E., Hewins R. H., Mittlefehldt D. W., Lindstrom M. M., Xiao X., and Lipschutz M. E. (1992) Meteoritics, submitted. 6. Xiao X. and Lipschutz M. E. (1992) J. Geophys. Res. Planets, in press. 7. Paul R. L. and Lipschutz M. E. (1989) Z. Naturf. 44a, 978-987. 8. Paul R. L. and Lipschutz M. E. (1990) Geochim. Cosmochim. Acta 54, 3185-3195.

  20. Susceptibility of Skeletal Muscle to Coxsackie A2 Virus Infection: Effects of Botulinum Toxin and Denervation

    NASA Astrophysics Data System (ADS)

    Andrew, Clifford G.; Drachman, Daniel B.; Pestronk, Alan; Narayan, Opendra

    1984-02-01

    Coxsackie A viruses can infect denervated but not innervated mature skeletal muscles. The role of synaptic transmission in preventing susceptibility to Coxsackievirus infection was studied by surgically denervating leg muscles of mice or injecting the muscles with botulinum toxin to block quantal release of acetylcholine. Control muscles were injected with heat-inactivated toxin. Subsequent injection of Coxsackie A2 virus resulted in extensive virus replication and tissue destruction in the denervated and botulinum toxin-treated muscles, while the control muscles showed only minimal changes. This suggests that the susceptibility of skeletal muscle to Coxsackievirus infection is regulated by synaptic transmission.

  1. Vaccination with plasmid DNA encoding a mutant toxic shock syndrome toxin-1 ameliorates toxin-induced lethal shock in mice.

    PubMed

    Feng, Mao-Hui; Cui, Jing-Chun; Nakane, Akio; Hu, Dong-Liang

    2013-01-01

    Staphylococcal toxic shock syndrome toxin-1 (TSST-1), a superantigenic toxin produced by Staphylococcus (S.) aureus, is a major cause of septic shock and toxic shock syndrome. To investigate whether vaccination with a plasmid DNA encoding a non-toxic mutant TSST-1 (mTSST-1) can protect mice against wild-type TSST-1-induced lethal shock, the mice were intranasally immunized with the plasmid DNA (named pcDNA-mTSST-1) plus a mucosal adjuvant, a non-toxic mutant labile toxin (mLT). After the immunization, the mice were challenged with TSST-1 and lipopolysaccharide (LPS). The survival rate of mice immunized with pcDNA-mTSST-1 plus mLT was higher than that of the control mice immunized with PBS alone, mLT alone, pcDNA-mTSST-1 alone, or a parent plasmid plus mLT. The titers of interferon-? (IFN-?) in the sera of mice immunized with pcDNA-mTSST-1 plus mLT were significantly lower than those of the mLT control mice. Immunization with pcDNA-mTSST-1 plus mLT increased the serum levels of TSST-1-specific antibodies, especially immunoglobulin G1 (IgG1) and IgG2a subclasses. Furthermore, the sera obtained from mice immunized with pcDNA-mTSST-1 plus mLT significantly inhibited the TSST-1-induced secretion of IFN-? and tumor necrosis factor-? (TNF-?) in murine spleen cells in vitro. These results indicate that immunization with pcDNA-mTSST-1 plus mLT provides protection against the lethal toxic shock of mice induced by wild-type TSST-1. The protective effect could be due to TSST-1-specific neutralizing antibodies as well as the inhibition of IFN-? and TNF-? secretions. Since TSST-1 is commonly released by invasive S. aureus, the pcDNA-mTSST-1 should be useful in preventing toxin-induced shock resulting from S. aureus infection. PMID:23985881

  2. Differential priming of soil carbon driven by soil depth and root impacts on carbon lability

    NASA Astrophysics Data System (ADS)

    De Graaff, M.; Jastrow, J. D.; Gilette, S.; Johns, A.; Wullschleger, S. D.

    2012-12-01

    An increase in root-derived labile soil carbon (C) inputs (e.g. exudates) can stimulate decomposition of more recalcitrant soil organic carbon (SOC) by priming microbial activity, which can lead to a net loss of soil C storage. This priming effect can be small or large and negative or positive, but the mechanisms by which root-C controls the magnitude and direction of SOC decomposition remain poorly understood. With this study we evaluated how small versus large differences in labile soil C availability affect microbial processing of simulated root exudate inputs and decomposition of SOC. We conducted a laboratory incubation experiment (60 days) with soils collected from under six switchgrass (Panicum virgatum) cultivars to a depth of 60 cm. Differences in specific root length among cultivars were expected to result in small differences in labile soil C availability, whereas differences associated with soil depth were expected to result in large differences in labile soil C availability. Soil cores were divided into 0-10 cm, 20-30 cm and 40-60 cm depth increments, and soils of all cultivars (no roots) across all three depths were incubated with addition of either: (1) water (60% water holding capacity), or (2) labile C provided as a 13C-labeled synthetic root exudate cocktail. We measured CO2 respiration throughout the experiment. The switchgrass was grown for three years in soils that formerly supported C3 pasture grasses, and the natural difference in 13C signature between C3 and C4 plants enabled quantification of differences in the lability of root-derived C among cultivars. Moreover the 13C labeled synthetic root-exudate cocktail amendment to soils allowed us to assess impacts of exudates-C addition on priming in soil derived from the different cultivars and from different depths. Our experiment led to three main results: (1) different cultivars of switchgrass regulate labile C availability across the soil profile differently; (2) small differences in labile C among the soils derived from different cultivars did not significantly mediate the impact of exudates-C additions on priming; (3) but, large differences in labile soil C contents among depths led to differences in priming effects, where greater priming effects were observed for shallow relative to deep soils across all days of the experiment. These results suggest that increased root-derived C inputs will have marginal impacts on decomposition of more stable SOC at depth and that their impact on priming will be similar in soils with small differences in available soil C.

  3. Mzm1 influences a labile pool of mitochondrial zinc important for respiratory function.

    PubMed

    Atkinson, Aaron; Khalimonchuk, Oleh; Smith, Pamela; Sabic, Hana; Eide, David; Winge, Dennis R

    2010-06-18

    Zinc is essential for function of mitochondria as a cofactor for several matrix zinc metalloproteins. We demonstrate that a labile cationic zinc component of low molecular mass exists in the yeast mitochondrial matrix. This zinc pool is homeostatically regulated in response to the cellular zinc status. This pool of zinc is functionally important because matrix targeting of a cytosolic zinc-binding protein reduces the level of labile zinc and interferes with mitochondrial respiratory function. We identified a series of proteins that modulate the matrix zinc pool, one of which is a novel conserved mitochondrial protein designated Mzm1. Mutant mzm1Delta cells have reduced total and labile mitochondrial zinc, and these cells are hypersensitive to perturbations of the labile pool. In addition, mzm1Delta cells have a destabilized cytochrome c reductase (Complex III) without any effects on Complexes IV or V. Thus, we have established that a link exists between Complex III integrity and the labile mitochondrial zinc pool. PMID:20404342

  4. Development of pH-sensitive self-nanoemulsifying drug delivery systems for acid-labile lipophilic drugs.

    PubMed

    Zhao, Tianjing; Maniglio, Devid; Chen, Jie; Chen, Bin; Migliaresi, Claudio

    2016-03-01

    Oral administration is the most convenient way of all the drug delivery routes. Orally administered bioactive compounds must resist the harsh acidic fluids or enzyme digestion in stomach, to reach their absorbed destination in small intestine. This is the case for silibinin, a drug used to protect liver cells against toxins that has also been demonstrated in vitro to possess anti-cancer effects. However, as many other drugs, silibinin can degrade in the stomach due to the action of the gastric fluid. The use of pH-sensitive self-nanoemulsifying drug delivery systems (pH-SNEDDS) could overcome the drawback due to degradation of the drug in the stomach while enhancing its solubility and dissolution rate. In this paper we have investigated pH-sensitive self-nanoemulsifying formulations containing silibinin as model drug. Pseudo-ternary phase diagrams have been constructed in order to identify the self-emulsification regions under different pH. Solubility of silibinin in selected formulations has been assessed and stability of the pure drug and of the silibinin loaded pH-SNEDDS formulations in simulated gastric fluid had been compared. Droplet size of the optimized pH-SNEDDS has been correlated to pH, volume of dilution medium and silibinin loading amount. TEM (transmission electron microscopy) studies have shown that emulsion droplets had spherical shape and narrow size distribution. In vitro drug release studies of the optimal pH-SNEDDS indicated substantial increase of the drug release and release rate in comparison to pure silibinin and to the commercial silibinin tablet. The results indicated that pH-SNEDDS have potential to improve the biopharmaceutics properties of acid-labile lipophilic drugs. PMID:26923270

  5. Sodium Channel Inhibiting Marine Toxins

    NASA Astrophysics Data System (ADS)

    Llewellyn, Lyndon E.

    Saxitoxin (STX), tetrodotoxin (TTX) and their many chemical relatives are part of our daily lives. From killing people who eat seafood containing these toxins, to being valuable research tools unveiling the invisible structures of their pharmacological receptor, their global impact is beyond measure. The pharmacological receptor for these toxins is the voltage-gated sodium channel which transports Na ions between the exterior to the interior of cells. The two structurally divergent families of STX and TTX analogues bind at the same location on these Na channels to stop the flow of ions. This can affect nerves, muscles and biological senses of most animals. It is through these and other toxins that we have developed much of our fundamental understanding of the Na channel and its part in generating action potentials in excitable cells.

  6. EVALUATION OF DEVELOPMENTAL TOXICITY OF ALGAL TOXINS

    EPA Science Inventory

    The Office of Water is responsible for regulating waterborne pathogens and chemicals in drinking water. Algal toxins are present in some water sources but their reproductive risks are undetermined. Therefore, Algal toxins are being examined for potential developmental toxicity ...

  7. The 1.9 A crystal structure of heat-labile shrimp alkaline phosphatase.

    PubMed

    de Backer, Maaike; McSweeney, Sean; Rasmussen, Hanne B; Riise, Bjrn W; Lindley, Peter; Hough, Edward

    2002-05-17

    Alkaline phosphatases are non-specific phosphomonoesterases that are distributed widely in species ranging from bacteria to man. This study has concentrated on the tissue-nonspecific alkaline phosphatase from arctic shrimps (shrimp alkaline phosphatase, SAP). Originating from a cold-active species, SAP is thermolabile and is used widely in vitro, e.g. to dephosphorylate DNA or dNTPs, since it can be inactivated by a short rise in temperature. Since alkaline phosphatases are zinc-containing enzymes, a multiwavelength anomalous dispersion (MAD) experiment was performed on the zinc K edge, which led to the determination of the structure to a resolution of 1.9 A. Anomalous data clearly showed the presence of a zinc triad in the active site, whereas alkaline phosphatases usually contain two zinc and one magnesium ion per monomer. SAP shares the core, an extended beta-sheet flanked by alpha-helices, and a metal triad with the currently known alkaline phosphatase structures (Escherichia coli structures and a human placental structure). Although SAP lacks some features specific for the mammalian enzyme, their backbones are very similar and may therefore be typical for other higher organisms. Furthermore, SAP possesses a striking feature that the other structures lack: surface potential representations show that the enzyme's net charge of -80 is distributed such that the surface is predominantly negatively charged, except for the positively charged active site. The negatively charged substrate must therefore be directed strongly towards the active site. It is generally accepted that optimization of the electrostatics is one of the characteristics related to cold-adaptation. SAP demonstrates this principle very clearly. PMID:12083516

  8. The polychaete worm Nereis diversicolor increases mercury lability and methylation in intertidal mudflats.

    PubMed

    Sizmur, Tom; Canrio, Joo; Edmonds, Samuel; Godfrey, Adam; O'Driscoll, Nelson J

    2013-08-01

    The polychaete worm Nereis diversicolor engineers its environment by creating oxygenated burrows in anoxic intertidal sediments. The authors carried out a laboratory microcosm experiment to test the impact of polychaete burrowing and feeding activity on the lability and methylation of mercury in sediments from the Bay of Fundy, Canada. The concentration of labile inorganic mercury and methylmercury in burrow walls was elevated compared to worm-free sediments. Mucus secretions and organic detritus in worm burrows increased labile mercury concentrations. Worms decreased sulfide concentrations, which increased Hg bioavailability to sulfate-reducing bacteria and increased methylmercury concentrations in burrow linings. Because the walls of polychaete burrows have a greater interaction with organisms, and the overlying water, the concentrations of mercury and methylmercury they contain is more toxicologically relevant to the base of a coastal food web than bulk samples. The authors recommend that researchers examining Hg in marine environments account for sediment dwelling invertebrate activity to more fully assess mercury bioavailability. PMID:23633443

  9. Mood lability among offspring of parents with bipolar disorder and community controls

    PubMed Central

    Birmaher, Boris; Goldstein, Benjamin I; Axelson, David A; Monk, Kelly; Hickey, Mary Beth; Fan, Jieyu; Iyengar, Satish; Ha, Wonho; Diler, Rasim S; Goldstein, Tina; Brent, David; Ladouceur, Cecile D; Sakolsky, Dara; Kupfer, David J

    2013-01-01

    Objectives Early identification of bipolar disorder (BP) symptomatology is crucial for improving the prognosis of this illness. Increased mood lability has been reported in BP. However, mood lability is ubiquitous across psychiatric disorders and may be a marker of severe psychopathology and not specific to BP. To clarify this issue, this study examined the prevalence of mood lability and its components in offspring of BP parents and offspring of community control parents recruited through the Pittsburgh Bipolar Offspring Study. Methods Forty-one school-age BP offspring of 38 BP parents, 257 healthy or non-BP offspring of 174 BP parents, and 192 offspring of 117 control parents completed a scale that was developed to evaluate mood lability in youth, i.e., the Childrens Affective Lability Scale (CALS). Results A factor analysis of the parental CALS, and in part the child CALS, revealed Irritability, Mania, and Anxiety/Depression factors, with most of the variance explained by the Irritability factor. After adjusting for confounding factors (e.g., parental and offspring non-BP psychopathology), BP offspring of BP parents showed the highest parental and child total and factor scores, followed by the non-BP offspring of BP parents, and then the offspring of the controls. Conclusions Mood lability overall and mania-like, anxious/depressed, and particularly irritability symptoms may be a prodromal phenotype of BP among offspring of parents with BP. Prospective studies are warranted to clarify whether these symptoms will predict the development of BP and/or other psychopathology. If confirmed, these symptoms may become a target of treatment and biological studies before BP develops. PMID:23551755

  10. Biotic and abiotic controls on diurnal fluctuations in labile soil phosphorus of a wet tropical forest.

    PubMed

    Vandecar, Karen L; Lawrence, Deborah; Wood, Tana; Oberbauer, Steven F; Das, Rishiraj; Tully, Katherine; Schwendenmann, Luitgard

    2009-09-01

    The productivity of many tropical wet forests is generally limited by bioavailable phosphorus (P). Microbial activity is a key regulator of P availability in that it determines both the supply of P through organic matter decomposition and the depletion of bioavailable P through microbial uptake. Both microbial uptake and mineralization occur rapidly, and their net effect on P availability varies with soil moisture, temperature, and soil organic matter quantity and quality. Exploring the mechanisms driving P availability at fine temporal scales can provide insight into the coupling of carbon, water, and nutrient cycles, and ultimately, the response of tropical forests to climate change. Despite the recognized importance of P cycling to the dynamics of wet tropical forests and their potential sensitivity to short-term fluctuations in bioavailable P, the diurnal pattern of P remains poorly understood. This study quantifies diurnal fluctuations in labile soil P and evaluates the importance of biotic and abiotic factors in driving these patterns. To this end, measurements of labile P were made every other hour in a Costa Rican wet tropical forest oxisol. Spatial and temporal variation in Bray-extractable P were investigated in relation to ecosystem carbon flux, soil CO2 efflux, soil moisture, soil temperature, solar radiation, and sap-flow velocity. Spatially averaged bi-hourly (every two hours) labile P ranged from 0.88 to 2.48 microg/g across days. The amplitude in labile P throughout the day was 0.61-0.82 microg/g (41-54% of mean P concentrations) and was characterized by a bimodal pattern with a decrease at midday. Labile P increased with soil CO2 efflux and soil temperature and declined with increasing sap flow and solar radiation. Together, soil CO2 efflux, soil temperature, and sap flow explained 86% of variation in labile P. PMID:19769132

  11. Labile carbon concentrations are strongly linked to plant production in Arctic tussock tundra soils

    NASA Astrophysics Data System (ADS)

    Darrouzet-Nardi, A.; Weintraub, M. N.; Euskirchen, E. S.; Steltzer, H.; Sullivan, P.

    2013-12-01

    The exchange of carbon and nutrients between plants and microbes is a key determinant of carbon balance in Arctic soils. Microbes rely on labile plant carbon for the energy they need to produce enzymes that can release nutrients and less energetically favorable carbon from soil organic matter. One of the main mechanisms of carbon transfer is rhizodeposition, the exudation of labile plant carbon such as sugars from roots into the rhizosphere. Despite the importance of this flow of energy and materials from plants to microbes, there have been few attempts to quantify labile carbon pools or fluxes in Arctic soils. To improve our knowledge of labile carbon dynamics in Arctic soils, we address two basic questions: (1) What are the seasonal patterns of labile carbon concentrations? and (2) How do seasonal patterns in labile carbon correlate with plant production, microbial biomass, and soil nutrients? We measured concentrations of total reducing sugars (TRS) in the soil solution of moist acidic tussock tundra on 28 dates during the 2012 growing season in 20 plots of an early snowmelt warming experiment. We evaluated these total reducing sugar concentrations in the context of eddy flux carbon exchange data, plant NDVI, total dissolved carbon in soils, microbial biomass, and soil nutrients. Though we did not see treatment effects of the snowmelt warming experiment, we did observe a clear seasonal pattern in TRS concentrations in which they started low at the time of thaw, then built to a maximum value around the time of peak plant physiology in July, followed by a decline as plants senesced. We observed a clear correlation between TRS and gross primary production (GPP). NDVI values also increased with TRS concentrations during the first half of the season and then leveled off as TRS began its decline. These relationships were in contrast to labile N concentrations, which remained at low concentrations all season. Our data suggest that rhizodeposition of labile carbon compounds in Arctic tundra ecosystems has a strong seasonal pattern that closely tracks plant production. This connection suggests that if plant production increases in a warmer Arctic, plants may ease carbon limitation to summer microbial activity.

  12. Seasonal changes and biochemical composition of the labile organic matter flux in the Cretan Sea

    NASA Astrophysics Data System (ADS)

    Danovaro, Roberto; Della Croce, Norberto; Dell'Anno, Antonio; Mauro Fabiano; Marrale, Daniela; Martorano, Daniela

    2000-08-01

    Downward fluxes of labile organic matter (lipids, proteins and carbohydrates) at 200 (trap A) and 1515 m depth (trap B), measured during a 12 months sediment trap experiment, are presented, together with estimates of the bacterial and cyanobacterial biomasses associated to the particles. The biochemical composition of the settling particles was determined in order to provide qualitative and quantitative information on the flux of readily available organic carbon supplying the deep-sea benthic communities of the Cretan Sea. Total mass flux and labile carbon fluxes were characterised by a clear seasonality. Higher labile organic fluxes were reported in trap B, indicating the presence of resuspended particles coming from lateral inputs. Particulate carbohydrates were the major component of the flux of labile compounds (on annual average about 66% of the total labile organic flux) followed by lipids (20%) and proteins (13%). The biopolymeric carbon flux was very low (on annual average 0.9 and 1.2 gC m -2 y -1, at trap A and B). Labile carbon accounted for most of the OC flux (on annual average 84% and 74% in trap A and B respectively). In trap A, highest carbohydrate and protein fluxes in April and September, corresponded to high faecal pellet fluxes. The qualitative composition of the organic fluxes indicated a strong protein depletion in trap B and a decrease of the bioavailability of the settling particles as a result of a higher degree of dilution with inorganic material. Quantity and quality of the food supply to the benthos displayed different temporal patterns. Bacterial biomass in the sediment traps (on average 122 and 229 μgC m -2 d -1 in trap A and B, respectively) was significantly correlated to the flux of labile organic carbon, and particularly to the protein and carbohydrate fluxes. Cyanobacterial flux (on average, 1.1 and 0.4 μgC m -2 d -1, in trap A and B, respectively) was significantly correlated with total mass and protein fluxes only in trap A. Bacterial carbon flux, equivalent to 84.2 and 156 mgC m -2 y -1, accounted for 5-6.5% of the labile carbon flux (in trap A and B respectively) and for 22-41% protein pool of the settling particles. These results suggest that in the Cretan Sea, bacteria attached to the settling particles represent a potential food source of primary importance for deep-sea benthic communities.

  13. MCEARD - CYANOBACTERIA AND THEIR TOXINS

    EPA Science Inventory

    Harmful algal blooms (HAB) of cyanobacteria, also known as blue-green algae, have recently become more spatially and temporally prevalent in the US and worldwide. Waterborne cyanobacteria and their highly potent toxins are a significant hazard for human health and the ecosystem....

  14. Cholera toxin notches epithelial junctions.

    PubMed

    Lemichez, Emmanuel; Stefani, Caroline

    2013-09-11

    Cholera toxin (CT) is the factor responsible for watery diarrhea associated with Vibrio cholerae infection. In this issue, Guichard et al. (2013) report that CT compromises intestinal epithelium barrier function via cyclic AMP (cAMP)-induced disruption of Rab11- and exocyst-dependent delivery of endocytic recycling cargo to cell-cell junctions. PMID:24034608

  15. Risk Assessment of Shellfish Toxins

    PubMed Central

    Munday, Rex; Reeve, John

    2013-01-01

    Complex secondary metabolites, some of which are highly toxic to mammals, are produced by many marine organisms. Some of these organisms are important food sources for marine animals and, when ingested, the toxins that they produce may be absorbed and stored in the tissues of the predators, which then become toxic to animals higher up the food chain. This is a particular problem with shellfish, and many cases of poisoning are reported in shellfish consumers each year. At present, there is no practicable means of preventing uptake of the toxins by shellfish or of removing them after harvesting. Assessment of the risk posed by such toxins is therefore required in order to determine levels that are unlikely to cause adverse effects in humans and to permit the establishment of regulatory limits in shellfish for human consumption. In the present review, the basic principles of risk assessment are described, and the progress made toward robust risk assessment of seafood toxins is discussed. While good progress has been made, it is clear that further toxicological studies are required before this goal is fully achieved. PMID:24226039

  16. Toxins as Weapons: A Historical Review.

    PubMed

    Pita, R; Romero, A

    2014-07-01

    This review article summarizes the use of toxins as weapons dating from the First World War until today, when there is a high concern of possible terrorist attacks with weapons of mass destruction. All through modern history, military programs and terrorist groups have favored toxins because of their high toxicity. However, difficulties of extraction or synthesis, as well as effective dissemination to cause a large number of casualties, have been the most important drawbacks. Special emphasis is focused on ricin and botulinum toxin, the most important toxins that have attracted the attention of military programs and terrorist groups. Other toxins like trichothecenes, saxitoxin, and Staphylococcal enterotoxin B (SEB) are also discussed. A short section about anthrax is also included: Although Bacillus anthracis is considered a biological weapon rather than a toxin weapon, it produces a toxin that is finally responsible for the anthrax disease. PMID:26227025

  17. Antibody-based bacterial toxin detection

    NASA Astrophysics Data System (ADS)

    Menking, Darrell E.; Heitz, Jonathon M.; Anis, Nabil A.; Thompson, Roy G.

    1994-03-01

    Fiber optic evanescent fluorosensors are under investigation in our laboratory for the study of drug-receptor interactions for detection of threat agents and antibody-antigen interactions for detection of biological toxins. In a one step assay, antibodies against Cholera toxin or Staphylococcus Enterotoxin B were noncovalently immobilized on quartz fibers and probed with fluorescein-isothiocyanate (FITC)-labeled toxins. In the two-step assay, Cholera toxin or Botulinum toxoid A was immobilized onto the fiber, followed by incubation in an antiserum or partially purified antitoxin IgG. These were then probed with FITC-anti-IgG antibodies. Unlabeled toxins competed with labeled toxins or antitoxin IgG in a dose-dependent manner and the detection of the toxins was in the nanomolar range.

  18. Nemertean toxin genes revealed through transcriptome sequencing.

    PubMed

    Whelan, Nathan V; Kocot, Kevin M; Santos, Scott R; Halanych, Kenneth M

    2014-12-01

    Nemerteans are one of few animal groups that have evolved the ability to utilize toxins for both defense and subduing prey, but little is known about specific nemertean toxins. In particular, no study has identified specific toxin genes even though peptide toxins are known from some nemertean species. Information about toxin genes is needed to better understand evolution of toxins across animals and possibly provide novel targets for pharmaceutical and industrial applications. We sequenced and annotated transcriptomes of two free-living and one commensal nemertean and annotated an additional six publicly available nemertean transcriptomes to identify putative toxin genes. Approximately 63-74% of predicted open reading frames in each transcriptome were annotated with gene names, and all species had similar percentages of transcripts annotated with each higher-level GO term. Every nemertean analyzed possessed genes with high sequence similarities to known animal toxins including those from stonefish, cephalopods, and sea anemones. One toxin-like gene found in all nemerteans analyzed had high sequence similarity to Plancitoxin-1, a DNase II hepatotoxin that may function well at low pH, which suggests that the acidic body walls of some nemerteans could work to enhance the efficacy of protein toxins. The highest number of toxin-like genes found in any one species was seven and the lowest was three. The diversity of toxin-like nemertean genes found here is greater than previously documented, and these animals are likely an ideal system for exploring toxin evolution and industrial applications of toxins. PMID:25432940

  19. Similarity of Monozygotic and Dizygotic Twins in Level and Lability of Subclinically Depressed Mood.

    ERIC Educational Resources Information Center

    Wierzbicki, Michael

    1986-01-01

    Ninety-two adult twin-pairs were recruited. Twin zygosity was determined by self-report inventory. Monozygotic twins resembled one another more than dizygotic twins in most measures of both level and lability of mood, which provides modest evidence for a genetic influence on subclinical levels of depression. (Author/ABB)

  20. Labile compounds in plant litter reduce the sensitivity of decomposition to warming and altered precipitation.

    PubMed

    Suseela, Vidya; Tharayil, Nishanth; Xing, Baoshan; Dukes, Jeffrey S

    2013-10-01

    Together, climate and litter quality strongly regulate decomposition rates. Although these two factors and their interaction have been studied across species in continent-scale experiments, few researchers have studied how labile and recalcitrant compounds interact to influence decomposition, or the climate sensitivity of decomposition, within a litter type. Over a period of 3 yr, we studied the effects of warming and altered precipitation on mass loss and compound-specific decomposition using two litter types that possessed similar heteropolymer chemistry, but different proportions of labile and recalcitrant compounds. Climate treatments immediately affected the mass loss of the more recalcitrant litter, but affected the more labile litter only after 2 yr. After 3 yr, although both litter types had lost similar amounts of mass, warming (c. 4°C) and supplemental precipitation (150% of ambient) together accelerated the degradation of alkyl-carbon and lignin only in the more recalcitrant litter, highlighting the role of initial litter quality in determining whether the chemistry of litter residues converges or diverges under different climates. Our finding that labile compounds in litter reduce the climate sensitivity of mass loss and the decomposition of recalcitrant matrix is novel. Our results highlight the potential for litter quality to regulate the effect of climatic changes on the sequestration of litter-derived carbon. PMID:23822593

  1. Glycosylation efficiencies on different solid supports using a hydrogenolysis-labile linker.

    PubMed

    Collot, Mayeul; Eller, Steffen; Weishaupt, Markus; Seeberger, Peter H

    2013-01-01

    Automated oligosaccharide assembly requires suitable linkers to connect the first monosaccharide to a solid support. A new hydrogenolysis-labile linker that is stable under both acidic and basic conditions was designed, synthesized and coupled to different resins. Glycosylation and cleavage efficiencies on these functionalized solid supports were investigated, and restrictions for the choice of solid support for oligosaccharide synthesis were found. PMID:23400514

  2. Solubility and lability of cadmium and zinc in two soils treated with organic matter

    SciTech Connect

    Almaas, A.R.; McBride, M.B.; Singh, B.R.

    2000-03-01

    The effect of organic matter (pig manure, sus scrofa) addition on solubility and free Cd(II) and Zn(II) speciation was studied in two mineral soils. The soils were extracted with ultra pure 0.01 M KNO{sub 3}, and the extracts were analyzed for total dissolved Cd and Zn by graphite furnace AAS and ICP, respectively, and for labile Cd and Zn by differential pulse anodic stripping voltammetry (DPASV). Based on the assumption that the non-ASV-labile fraction of the total dissolved Cd and Zn was organically bound to fulvic acid (FA), the relative fraction of labile to total dissolved Cd and Zn was used to estimate conditional stability constants (log K) for the formation of Cd-Fulvate (CdFA) and Zn-Fulvate (ZnFA). Species of organically and inorganically associated Cd and Zn, as affected by the addition of pig manure to the two different soil types, were calculated. The addition of organic matter increased the solubility of Cd and Zn in both soils by the formation of organo-metallic complexes. The lability of Zn was, however, reduced substantially, whereas for Cd it was unaffected. The conditional log K values calculated indicate that the stability of organo-metallic complexes with Cd and Zn may be more important than reported previously. This implies that increasing concentrations of dissolved organic acids can increase their solubility, thus leading to the leaching of Cd and Zn into ground water.

  3. Non-labile silver species in biosolids remain stable throughout 50 years of weathering and ageing.

    EPA Science Inventory

    Increasing commercial use of nanosilver has focussed attention on the fate of silver (Ag) in the wastewater release pathway. This paper reports the speciation and lability of Ag in archived, stockpiled, and contemporary biosolids from the UK, USA and Australia, and indicates that...

  4. Enzymatically- and Ultraviolet-labile Phosphorus in Humic Acid Fractions From Rice Soils

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Humic acid is an important soil component which can improve nutrient availability and impact other important chemical, biological, and physical properties of soils. We investigated the lability of phosphorus (P) in the mobile humic acid (MHA) and calcium humate (CaHA) fractions of four rice soils as...

  5. Affective lability and affect intensity as core dimensions of bipolar disorders during euthymic period.

    PubMed

    Henry, Chantal; Van den Bulke, Donatienne; Bellivier, Frank; Roy, Isabelle; Swendsen, Jol; M'Balara, Katia; Siever, Larry J; Leboyer, Marion

    2008-05-30

    Bipolar disorders are usually defined by alternative mood states, but a more precise characterization of the euthymic period could provide further insights into the pathophysiology of bipolar disorders. Surprisingly, few studies have investigated core affective dimensions in euthymic bipolar patients. In this study, we assessed 179 euthymic bipolar patients (score<12 on the Montgomery-Asberg Depression Scale and a score<6 on the Bech-Rafaelsen Mania Scale) compared with 86 control subjects using French versions of the Affective Lability Scale (ALS) and the Affect Intensity Measure (AIM). Data were analyzed by logistic regression. Our results showed that euthymic bipolar subjects reported having more intense emotions than controls and also had a higher affective lability. High scores in both affective dimensions were associated with a significantly higher risk for psychiatric axis I comorbidity. Moreover, a high affective lability score was associated with an earlier age of onset for bipolar disease. Affective lability and affect intensity might be two core dimensions of bipolar disorder during euthymic periods, suggesting that bipolar disorder is not circumscribed to mood episodes but also affects emotional reactivity between episodes. Both dimensions could account for the stress reactivity of bipolar patients that may lead to relapses. PMID:18295902

  6. Dual selective iron chelating probes with a potential to monitor mitochondrial labile iron pools.

    PubMed

    Abbate, Vincenzo; Reelfs, Olivier; Kong, Xiaole; Pourzand, Charareh; Hider, Robert C

    2015-12-24

    Mitochondria-targeted peptides incorporating dual fluorescent and selective iron chelators have been designed as novel biosensors for the mitochondrial labile iron pool. The probes were demonstrated to specifically co-localize with mitochondria and their fluorescence emission was found to be sensitive to the presence of iron. PMID:26567874

  7. Non-labile silver species in biosolids remain stable throughout 50 years of weathering and ageing.

    PubMed

    Donner, E; Scheckel, K; Sekine, R; Popelka-Filcoff, R S; Bennett, J W; Brunetti, G; Naidu, R; McGrath, S P; Lombi, E

    2015-10-01

    Increasing commercial use of nanosilver has focussed attention on the fate of silver (Ag) in the wastewater release pathway. This paper reports the speciation and lability of Ag in archived, stockpiled, and contemporary biosolids from the UK, USA and Australia, and indicates that biosolids Ag concentrations have decreased significantly over recent decades. XANES revealed the importance of reduced-sulfur binding environments for Ag speciation in materials ranging from freshly produced sludge to biosolids weathered under ambient environmental conditions for more than 50 years. Isotopic dilution with (110 m)Ag showed that Ag was predominantly non-labile in both fresh and aged biosolids (13.7% mean lability), with E-values ranging from 0.3 to 60 mg/kg and 5 mM CaNO3 extractable Ag from 1.2 to 609 μg/kg (0.002-3.4% of the total Ag). This study indicates that at the time of soil application, biosolids Ag will be predominantly Ag-sulfides and characterised by low isotopic lability. PMID:26021819

  8. A Longitudinal Study of Emotion Regulation, Emotion Lability/Negativity, and Internalizing Symptomatology in Maltreated and Nonmaltreated Children

    PubMed Central

    Kim-Spoon, Jungmeen; Cicchetti, Dante; Rogosch, Fred A.

    2013-01-01

    The longitudinal contributions of emotion regulation and emotion lability/negativity to internalizing symptomatology were examined in a low-income sample (171 maltreated and 151 nonmaltreated children, from age 7 to 10 years). Latent difference score models indicated that, for both maltreated and nonmaltreated children, emotion regulation was a mediator between emotion lability/negativity and internalizing symptomatology, whereas emotion lability/negativity was not a mediator between emotion regulation and internalizing symptomatology. Early maltreatment was associated with high emotion lability/negativity (age 7) that contributed to poor emotion regulation (age 8), which in turn was predictive of increases in internalizing symptomatology (from age 8 to 9). The results imply important roles of emotion regulation in the development of internalizing symptomatology, especially for children with high emotion lability/negativity. PMID:23034132

  9. Oxygen consumption and labile dissolved organic carbon uptake by benthic biofilms

    NASA Astrophysics Data System (ADS)

    de Falco, Natalie; Boano, Fulvio; Arnon, Shai

    2015-04-01

    Biogeochemical activity in streams is often magnified at interfaces, such as in the case of biofilm growth near the surface of the stream sediments. The objective of this study was to evaluate the relative importance of surficial biofilms versus the biofilm in the hyporheic zone to the processes of biodegradation of a labile dissolved organic carbon (DOC) and to oxygen consumption. Experiments were conducted in a recirculating flume, equipped with a drainage system that enables the control on losing and gaining fluxes. A surficial biofilm was developed over a sandy streambed with dune-shaped bed forms, by providing labile DOC (sodium benzoate) and nitrate. Homogeneously distributed biofilm was obtained by the same feeding strategy but with mixing the sediments manually on a daily basis. After the biofilm growth period, transformation of the labile DOC under different overlying velocities and losing or gaining fluxes was studied after spiking with sodium benzoate and by monitoring the decrease in DOC concentration in the bulk water over time using an online UV/Vis spectrophotometer. In addition, oxygen profiles across the water-streambed interface were measured at different locations along the bed form using oxygen microelectrodes. Preliminary results showed that the rate of labile DOC degradation increased exponentially with increasing overlying water velocity, regardless of the type of biofilm. Gaining and losing conditions did not play a critical role in the DOC degradation regardless of the type of biofilm, because the labile DOC was quickly utilized close to the surface. Under losing conditions, complete depletion of oxygen was observed within the top 5 millimeters, regardless of the biofilm type. In contrast, oxygen profiles under gaining condition showed an incomplete consumption of oxygen followed by an increase in the concentration of oxygen deeper in the sediments due to the upward flow of oxygenated groundwater. The results suggest that the transformation of labile DOC occurs in the upper millimeters of the streambed, and the size and shape of the hyporheic flow paths are less important for aerobic activity. In addition, the effect of overlying water velocity on labile DOC transformation was shown to be more influential than losing and gaining fluxes.

  10. Primary production contributes to non-labile organic matter generation in the estuarine and coastal zone

    NASA Astrophysics Data System (ADS)

    Zhu, Z.; Zhang, J.; Wu, Y.

    2013-12-01

    Oceanic generation of refractory organic matter is an important pathway for safe and long time scale safe carbon sequestration. Since refractory/non-labile organic matter generation is highly related with microorganism, estuaries and coastal zones with high primary production should be important regions for such generation process. We investigated the particulate organic matter in the estuarine and adjacent coastal zone of the Changjiang (Yangtze River). Peptidoglycan estimated on the basis of D-form of amino acids enantiomers showed a large variation in the estuary but generally lower than the lower reaches (XLJ). Peptidoglycan quickly decreased from the river to the sea, when DI increased from negative to < 0.5. The decrease can be due to dilution by fresh organic matter and seawater. But when DI > 0.5, the peptidoglycan concentration began to positively relate with organic matter freshness and normalized peptidoglycan was comparable to or even higher than that in terrestrial organic matter. This indicates that estuarine and coastal zones make a significant contribution to non-labile organic matter production. Further analysis suggests that heterotrophic bacteria and Synechococcus are notable contributors. For large river's estuary and adjacent coastal zone, terrestrial inputs promote high in situ production. The generated fresh organic matter in the estuary further promotes heterotrophic bacteria. Since the generation of non-labile organic matter process is both contributed by autotrophic and heterotrophic microorganisms, primary production is indirectly generating refractory/non-labile organic matter. And the refractory/non labile organic matter production occurs routinely during every productive season. On another aspect, considering the shallow water depth (usually < 100 m) and high sedimentation rate (e.g., 0-5 cm/year for the Changjiang Estuary), the organic matter can be buried in sediment much more easily than it is in the open ocean.

  11. Soil-specific response functions of organic matter mineralization to the availability of labile carbon.

    PubMed

    Paterson, Eric; Sim, Allan

    2013-05-01

    Soil organic matter (SOM) mineralization processes are central to the functioning of soils in relation to feedbacks with atmospheric CO2 concentration, to sustainable nutrient supply, to structural stability and in supporting biodiversity. Recognition that labile C-inputs to soil (e.g. plant-derived) can significantly affect mineralization of SOM ('priming effects') complicates prediction of environmental and land-use change effects on SOM dynamics and soil C-balance. The aim of this study is to construct response functions for SOM priming to labile C (glucose) addition rates, for four contrasting soils. Six rates of glucose (3atm% (13) C) addition (in the range 0-1mg glucose g(-1) soil day(-1) ) were applied for 8days. Soil CO2 efflux was partitioned into SOM- and glucose-derived components by isotopic mass balance, allowing quantification of SOM priming over time for each soil type. Priming effects resulting from pool substitution effects in the microbial biomass ('apparent priming') were accounted for by determining treatment effects on microbial biomass size and isotopic composition. In general, SOM priming increased with glucose addition rate, approaching maximum rates specific for each soil (up to 200%). Where glucose additions saturated microbial utilization capacity (>0.5mg glucose g(-1) soil), priming was a soil-specific function of glucose mineralization rate. At low to intermediate glucose addition rates, the magnitude (and direction) of priming effects was more variable. These results are consistent with the view that SOM priming is supported by the availability of labile C, that priming is not a ubiquitous function of all components of microbial communities and that soils differ in the extent to which labile C stimulates priming. That priming effects can be represented as response functions to labile C addition rates may be a means of their explicit representation in soil C-models. However, these response functions are soil-specific and may be affected by several interacting factors at lower addition rates. PMID:23505211

  12. Effects of wetland recovery on soil labile carbon and nitrogen in the Sanjiang Plain.

    PubMed

    Huang, Jingyu; Song, Changchun; Nkrumah, Philip Nti

    2013-07-01

    Soil management significantly affects the soil labile organic factors. Understanding carbon and nitrogen dynamics is extremely helpful in conducting research on active carbon and nitrogen components for different kinds of soil management. In this paper, we examined the changes in microbial biomass carbon (MBC), microbial biomass nitrogen (MBN), dissolved organic carbon (DOC), and dissolved organic nitrogen (DON) to assess the effect and mechanisms of land types, organic input, soil respiration, microbial species, and vegetation recovery under Deyeuxia angustifolia freshwater marshes (DAMs) and recovered freshwater marsh (RFM) in the Sanjiang Plain, Northeast China. Identifying the relationship among the dynamics of labile carbon, nitrogen, and soil qualification mechanism using different land management practices is therefore important. Cultivation and land use affect intensely the DOC, DON, MBC, and MBN in the soil. After DAM soil tillage, the DOC, DON, MBC, and MBN at the surface of the agricultural soil layer declined significantly. In contrast, their recovery was significant in the RFM surface soil. A long time was needed for the concentration of cultivated soil total organic carbon and total nitrogen to be restored to the wetland level. The labile carbon and nitrogen fractions can reach a level similar to that of the wetland within a short time. Typical wetland ecosystem signs, such as vegetation, microbes, and animals, can be recovered by soil labile carbon and nitrogen fraction restoration. In this paper, the D. angustifolia biomass attained natural wetland level after 8years, indicating that wetland soil labile fractions can support wetland eco-function in a short period of time (4 to 8years) for reconstructed wetland under suitable environmental conditions. PMID:23151838

  13. Why do we study animal toxins?

    PubMed

    Zhang, Yun

    2015-07-18

    Venom (toxins) is an important trait evolved along the evolutionary tree of animals. Our knowledges on venoms, such as their origins and loss, the biological relevance and the coevolutionary patterns with other organisms are greatly helpful in understanding many fundamental biological questions, i.e., the environmental adaptation and survival competition, the evolution shaped development and balance of venoms, and the sophisticated correlations among venom, immunity, body power, intelligence, their genetic basis, inherent association, as well as the cost-benefit and trade-offs of biological economy. Lethal animal envenomation can be found worldwide. However, from foe to friend, toxin studies have led lots of important discoveries and exciting avenues in deciphering and fighting human diseases, including the works awarded the Nobel Prize and lots of key clinic therapeutics. According to our survey, so far, only less than 0.1% of the toxins of the venomous animals in China have been explored. We emphasize on the similarities shared by venom and immune systems, as well as the studies of toxin knowledge-based physiological toxin-like proteins/peptides (TLPs). We propose the natural pairing hypothesis. Evolution links toxins with humans. Our mission is to find out the right natural pairings and interactions of our body elements with toxins, and with endogenous toxin-like molecules. Although, in nature, toxins may endanger human lives, but from a philosophical point of view, knowing them well is an effective way to better understand ourselves. So, this is why we study toxins. PMID:26228472

  14. The toxin and antidote puzzle

    PubMed Central

    2011-01-01

    Insects carry out essential ecological functions, such as pollination, but also cause extensive damage to agricultural crops and transmit human diseases such as malaria and dengue fever. Advances in insect transgenesis are making it increasingly feasible to engineer genes conferring desirable phenotypes, and gene drive systems are required to spread these genes into wild populations. Medea provides one solution, being able to spread into a population from very low initial frequencies through the action of a maternally-expressed toxin linked to a zygotically-expressed antidote. Several other toxin-antidote combinations are imaginable that distort the offspring ratio in favor of a desired transgene, or drive the population towards an all-male crash. We explore two such systemsSemele, which is capable of spreading a desired transgene into an isolated population in a confined manner; and Merea, which is capable of inducing a local population crash when located on the Z chromosome of a Lepidopteron pest. PMID:21876382

  15. Neutralising Antibodies against Ricin Toxin

    PubMed Central

    Prigent, Julie; Panigai, Laetitia; Lamourette, Patricia; Sauvaire, Didier; Devilliers, Karine; Plaisance, Marc; Volland, Herv; Crminon, Christophe; Simon, Stphanie

    2011-01-01

    The Centers for Disease Control and Prevention have listed the potential bioweapon ricin as a Category B Agent. Ricin is a so-called A/B toxin produced by plants and is one of the deadliest molecules known. It is easy to prepare and no curative treatment is available. An immunotherapeutic approach could be of interest to attenuate or neutralise the effects of the toxin. We sought to characterise neutralising monoclonal antibodies against ricin and to develop an effective therapy. For this purpose, mouse monoclonal antibodies (mAbs) were produced against the two chains of ricin toxin (RTA and RTB). Seven mAbs were selected for their capacity to neutralise the cytotoxic effects of ricin in vitro. Three of these, two anti-RTB (RB34 and RB37) and one anti-RTA (RA36), when used in combination improved neutralising capacity in vitro with an IC50 of 31 ng/ml. Passive administration of association of these three mixed mAbs (4.7 g) protected mice from intranasal challenges with ricin (5 LD50). Among those three antibodies, anti-RTB antibodies protected mice more efficiently than the anti-RTA antibody. The combination of the three antibodies protected mice up to 7.5 hours after ricin challenge. The strong in vivo neutralising capacity of this three mAbs combination makes it potentially useful for immunotherapeutic purposes in the case of ricin poisoning or possibly for prevention. PMID:21633505

  16. YoeB toxin is activated during thermal stress

    PubMed Central

    Janssen, Brian D; Garza-Sánchez, Fernando; Hayes, Christopher S

    2015-01-01

    Type II toxin-antitoxin (TA) modules are thought to mediate stress-responses by temporarily suppressing protein synthesis while cells redirect transcription to adapt to environmental change. Here, we show that YoeB, a ribosome-dependent mRNase toxin, is activated in Escherichia coli cells grown at elevated temperatures. YoeB activation is dependent on Lon protease, suggesting that thermal stress promotes increased degradation of the YefM antitoxin. Though YefM is efficiently degraded in response to Lon overproduction, we find that Lon antigen levels do not increase during heat shock, indicating that another mechanism accounts for temperature-induced YefM proteolysis. These observations suggest that YefM/YoeB functions in adaptation to temperature stress. However, this response is distinct from previously described models of TA function. First, YoeB mRNase activity is maintained over several hours of culture at 42°C, indicating that thermal activation is not transient. Moreover, heat-activated YoeB does not induce growth arrest nor does it suppress global protein synthesis. In fact, E. coli cells proliferate more rapidly at elevated temperatures and instantaneously accelerate their growth rate in response to acute heat shock. We propose that heat-activated YoeB may serve a quality control function, facilitating the recycling of stalled translation complexes through ribosome rescue pathways. PMID:26147890

  17. YoeB toxin is activated during thermal stress.

    PubMed

    Janssen, Brian D; Garza-Sánchez, Fernando; Hayes, Christopher S

    2015-08-01

    Type II toxin-antitoxin (TA) modules are thought to mediate stress-responses by temporarily suppressing protein synthesis while cells redirect transcription to adapt to environmental change. Here, we show that YoeB, a ribosome-dependent mRNase toxin, is activated in Escherichia coli cells grown at elevated temperatures. YoeB activation is dependent on Lon protease, suggesting that thermal stress promotes increased degradation of the YefM antitoxin. Though YefM is efficiently degraded in response to Lon overproduction, we find that Lon antigen levels do not increase during heat shock, indicating that another mechanism accounts for temperature-induced YefM proteolysis. These observations suggest that YefM/YoeB functions in adaptation to temperature stress. However, this response is distinct from previously described models of TA function. First, YoeB mRNase activity is maintained over several hours of culture at 42°C, indicating that thermal activation is not transient. Moreover, heat-activated YoeB does not induce growth arrest nor does it suppress global protein synthesis. In fact, E. coli cells proliferate more rapidly at elevated temperatures and instantaneously accelerate their growth rate in response to acute heat shock. We propose that heat-activated YoeB may serve a quality control function, facilitating the recycling of stalled translation complexes through ribosome rescue pathways. PMID:26147890

  18. Bacterial protein toxins in human cancers.

    PubMed

    Rosadi, Francesca; Fiorentini, Carla; Fabbri, Alessia

    2016-02-01

    Many bacteria causing persistent infections produce toxins whose mechanisms of action indicate that they could have a role in carcinogenesis. Some toxins, like CDT and colibactin, directly attack the genome by damaging DNA whereas others, as for example CNF1, CagA and BFT, impinge on key eukaryotic processes, such as cellular signalling and cell death. These bacterial toxins, together with other less known toxins, mimic carcinogens and tumour promoters. The aim of this review is to fulfil an up-to-date analysis of toxins with carcinogenic potential that have been already correlated to human cancers. Bacterial toxins-induced carcinogenesis represents an emerging aspect in bacteriology, and its significance is increasingly recognized. PMID:26534910

  19. Clostridium difficile Toxins: Mediators of Inflammation

    PubMed Central

    Shen, Aimee

    2012-01-01

    Clostridium difficile is a significant problem in hospital settings as the most common cause of nosocomial diarrhea worldwide. C. difficile infections (CDIs) are characterized by an acute intestinal inflammatory response with neutrophil infiltration. These symptoms are primarily caused by the glucosylating toxins, TcdA and TcdB. In the past decade, the frequency and severity of CDIs have increased markedly due to the emergence of so-called hypervirulent strains that overproduce cytotoxic glucosylating toxins relative to historical strains. In addition, these strains produce a third toxin, binary toxin or C. difficile transferase (CDT), that may contribute to hypervirulence. Both the glucosylating toxins and CDT covalently modify target cell proteins to cause disassembly of the actin cytoskeleton and induce severe inflammation. This review summarizes our current knowledge of the mechanisms by which glucosylating toxins and CDT disrupt target cell function, alter host physiology and stimulate immune responses. PMID:22237401

  20. Bt Toxin Modification for Enhanced Efficacy

    PubMed Central

    Deist, Benjamin R.; Rausch, Michael A.; Fernandez-Luna, Maria Teresa; Adang, Michael J.; Bonning, Bryony C.

    2014-01-01

    Insect-specific toxins derived from Bacillus thuringiensis (Bt) provide a valuable resource for pest suppression. Here we review the different strategies that have been employed to enhance toxicity against specific target species including those that have evolved resistance to Bt, or to modify the host range of Bt crystal (Cry) and cytolytic (Cyt) toxins. These strategies include toxin truncation, modification of protease cleavage sites, domain swapping, site-directed mutagenesis, peptide addition, and phage display screens for mutated toxins with enhanced activity. Toxin optimization provides a useful approach to extend the utility of these proteins for suppression of pests that exhibit low susceptibility to native Bt toxins, and to overcome field resistance. PMID:25340556

  1. Phage display and Shiga toxin neutralizers.

    PubMed

    Bernedo-Navarro, Robert Alvin; Yano, Tomomasa

    2016-04-01

    The current work presents an overview of the use of phage display technology for the identification and characterization of potential neutralizing agents for Shiga toxins. The last major Shiga toxin-associated disease outbreak, which took place in Germany in 2011, showed the international community that Shiga toxins remain a serious threat to public health. This is also demonstrated by the lack of specific therapies against Shiga toxin-induced Hemolytic Uremic Syndrome (HUS). Since its inception, phage display technology has played a key role in the development of antigen-specific (poly)-peptides or antibody fragments with specific biological properties. Herein, we review the current literature regarding the application of phage display to identify novel neutralizing agents against Shiga toxins. We also briefly highlight reported discoveries of peptides and heavy chain antibodies (VHH fragments or nanobodies) that can neutralize the cellular damage caused by these potent toxins. PMID:26898657

  2. Modeling of toxin-antibody interaction and toxin transport toward the endoplasmic reticulum.

    PubMed

    Skakauskas, Vladas; Katauskis, Pranas

    2016-01-01

    A model for toxin-antibody interaction and toxin trafficking towards the endoplasmic-reticulum is presented. Antibody and toxin (ricin) initially are delivered outside the cell. The model involves: the pinocytotic (cellular drinking) and receptor-mediated toxin internalization modes from the extracellular into the intracellular domain, its exocytotic excretion from the cytosol back to the extracellular medium, the intact toxin retrograde transport to the endoplasmic reticulum, the anterograde toxin movement outward from the cell across the plasma membrane, the lysosomal toxin degradation, and the toxin clearance (removal from the system) flux. The model consists of a set of coupled PDEs. Using an averaging procedure, the model is reduced to a system of coupled ODEs. Both PDEs and ODEs systems are solved numerically. Numerical results are illustrated by figures and discussed. PMID:26306534

  3. The tool of microbial genomics research for interpreting the lability of permafrost carbon and potential greenhouse gas feedbacks at different scales of resolution.

    NASA Astrophysics Data System (ADS)

    Waldrop, M. P.; Machelprang, R.; Hultman, J.; Wickland, K. P.

    2012-12-01

    One quarter of the earth's terrestrial surface is underlain by permafrost, or perennially frozen soils. Permafrost soils contain approximately 25% to 50% of the total global soil carbon pool nearly double the atmospheric carbon (C) reservoir. Decomposition of this C by microorganisms may produce globally significant quantities of both carbon dioxide and methane. These processes provide a positive feedback between climate change and the altered biogeochemistry of northern ecosystems. The fate of carbon residing in thawing permafrost soils depends on a number of physical factors including the thermal properties of soils (which affect heat flow rates), its disturbance regime (controlling changes in physical properties), and hydrologic regime (where soil-water interactions can rapidly thaw permafrost). Yet the mechanism of soil organic matter decomposition and greenhouse gas production operates primarily through the microbial loop: growth, carbon and nutrient mineralization, electron transfer, and enzyme production. We tested whether molecular analysis of microbial communities can be utilized as an indicator of permafrost C lability and potential greenhouse gas production from permafrost soils across multiple temporal and spatial scales. For short term studies of lability we compared rates of C turnover in soil incubations to chemical indices of soil lability, soil enzymes, and the abundance of soil microbial populations. Permafrost soils for the incubation ranged from frozen peatlands to dry uplands and Pleistocene Yedoma. For analysis at the annual to decadal scale, we utilized a permafrost thaw gradient at the Bonanza Creek LTER near Fairbanks Alaska. At this gradient, a Black Spruce forest underlain by permafrost thawed to form a thermokarst bog <50 years ago. Over the short term (months), the lability of permafrost C is reflected in the chemistry of dissolved constituents of permafrost, and it is also reflected in the change in abundance of total soil bacteria, which use DOC for growth and energy. At the decadal scale, permafrost thaw will result in wholesale change in plant community composition and ecosystem function, thus relying on an entirely different interpretation of shifts in microbial community composition to gain insight into greenhouse gas production. At Bonanza Creek, there are large differences in microbial community composition between intact permafrost and thermokarst bog soils. Metagenomic and metatransciptome analysis of microbial communities reveal that changes in community composition and representative functional genes reflect the dominant greenhouse gas producing processes occurring in those environments. Results indicated that the relative abundance of microbial functional groups based upon functional genes (e.g. methanogens, nitrate reducers, sulfate reducers) mirror the geochemistry of the system and pathways of potential greenhouse gas production. These data help to understand linkages between the level of detail undertaken within -omics research and the temporal scale of biogeochemical processes under examination.

  4. Cellular and Systemic Effects of Anthrax Lethal Toxin and Edema Toxin

    PubMed Central

    Moayeri, Mahtab; Leppla, Stephen H.

    2009-01-01

    Anthrax lethal toxin (LT) and edema toxin (ET) are the major virulence factors of anthrax and can replicate the lethality and symptoms associated with the disease. This review provides an overview of our current understanding of anthrax toxin effects in animal models and the cytotoxicity (necrosis and apoptosis) induced by LT in different cells. A brief reexamination of early historic findings on toxin in vivo effects in the context of our current knowledge is also presented. PMID:19638283

  5. Natural toxins and their therapeutic potential.

    PubMed

    Kapoor, V K

    2010-03-01

    Plants have been extensively investigated for exploring their therapeutic potentials, but there are comparatively scanty reports on drugs derived from animal kingdom, except for hormones. During last decade, the toxins that are used for defense by the animals, have been isolated and found useful tools for physiological and pharmacological studies, besides giving valuable leads to drug development. Toxins with interesting results have been isolated from the venoms of snakes, scorpions, spiders, snails, lizards, frogs and fish. The present review describe about some toxins as drugs and their biological activities. Some fungal, bacterial and marine toxins have also been covered in this article. PMID:21046975

  6. Toxin Detection by Surface Plasmon Resonance

    PubMed Central

    Hodnik, Vesna; Anderluh, Gregor

    2009-01-01

    Significant efforts have been invested in the past years for the development of analytical methods for fast toxin detection in food and water. Immunochemical methods like ELISA, spectroscopy and chromatography are the most used in toxin detection. Different methods have been linked, e.g. liquid chromatography and mass spectrometry (LC-MS), in order to detect as low concentrations as possible. Surface plasmon resonance (SPR) is one of the new biophysical methods which enables rapid toxin detection. Moreover, this method was already included in portable sensors for on-site determinations. In this paper we describe some of the most common methods for toxin detection, with an emphasis on SPR. PMID:22573957

  7. Application of Botulinum Toxin in Pain Management

    PubMed Central

    2011-01-01

    Botulinum toxin has been used for the treatment of many clinical disorders by producing temporary skeletal muscle relaxation. In pain management, botulinum toxin has demonstrated an analgesic effect by reducing muscular hyperactivity, but recent studies suggest this neurotoxin could have direct analgesic mechanisms different from its neuromuscular actions. At the moment, botulinum toxin is widely investigated and used in many painful diseases such as myofascial syndrome, headaches, arthritis, and neuropathic pain. Further studies are needed to understand the exact analgesic mechanisms, efficacy and complications of botulinum toxin in chronic pain disorders. PMID:21390172

  8. Application of botulinum toxin in pain management.

    PubMed

    Sim, Woo Seog

    2011-03-01

    Botulinum toxin has been used for the treatment of many clinical disorders by producing temporary skeletal muscle relaxation. In pain management, botulinum toxin has demonstrated an analgesic effect by reducing muscular hyperactivity, but recent studies suggest this neurotoxin could have direct analgesic mechanisms different from its neuromuscular actions. At the moment, botulinum toxin is widely investigated and used in many painful diseases such as myofascial syndrome, headaches, arthritis, and neuropathic pain. Further studies are needed to understand the exact analgesic mechanisms, efficacy and complications of botulinum toxin in chronic pain disorders. PMID:21390172

  9. Mechanism of CO displacement from an unusually labile rhenium complex: an experimental and theoretical investigation.

    PubMed

    Muhammad, Sohail; Yempally, Veeranna; Anas, Muhammad; Moncho, Salvador; Kyran, Samuel J; Brothers, Edward N; Darensbourg, Donald J; Bengali, Ashfaq A

    2012-12-01

    The displacement of a CO ligand from an unusually labile rhenium carbonyl complex containing a bidentate carboxyaldehyde pyrrolyl ligand by PPh(3) and pyridine has been investigated. The reaction is found to proceed by an associative, preequilibrium mechanism. Theoretical calculations support the experimental data and provide a complete energetic profile for the reaction. While the Re-CO bond is found to be intrinsically weak in these complexes, it is postulated that the unusual lability of this species is due to the presence of a weak aldehyde Re-O link that can easily dissociate to open a coordination site on the metal center and accommodate an incoming ligand prior to CO loss. The resulting intermediate complex has been identified by IR spectroscopy. The presence of the hemilabile pyrrolyl ligand provides a lower-energy reaction channel for the release of CO and may be of relevance in the design of CO-releasing molecules. PMID:23153281

  10. Relationship between the lability of sediment-bound Cd and its bioaccumulation in edible oyster.

    PubMed

    Chakraborty, Parthasarathi; Ramteke, Darwin; Chakraborty, Sucharita; Chennuri, Kartheek; Bardhan, Pratirupa

    2015-11-15

    A linkage between Cd speciation in sediments and its bioaccumulation in edible oyster (Crassostrea sp.) from a tropical estuarine system was established. Bioaccumulation of Cd in edible oyster increased with the increasing lability and dissociation rate constants of Cd-sediment complexes in the bottom sediments. Total Cd concentration in sediment was not a good indicator of Cd-bioavailability. Increasing trace metal competition in sediments increased lability and bioavailability of Cd in the tropical estuarine sediment. Low thermodynamic stability and high bioavailability of Cd in the estuarine sediment were responsible for high bioaccumulation of Cd in edible oysters (3.2-12.2mgkg(-1)) even though the total concentration of Cd in the bottom sediment was low (0.17-0.49mgkg(-1)). PMID:26359116

  11. Monitoring of labile zinc in cultures of Skeletonema costatum using a salt groundwater.

    PubMed

    Schintu, M; Koussih, L; Chevolot, L; Amiard, J C; Robert, J M

    1999-03-01

    Labile Zn concentration was monitored by differential pulse anodic stripping voltammetry (DPASV) throughout the exponential growth phase of the marine diatom Skeletonema costatum (Grev.) Cleve. Algal blooms were induced both under natural conditions and in laboratory experiments using a salt groundwater (salinity 33) from the Bay of Bourgneuf, northwest coast of France. Salt groundwater is a very complex medium containing high concentrations of dissolved organic matter and other trace metal adsorbents, such as phosphate, iron oxyhydroxides, and manganese and silicon oxides, which can bind metal ions, reducing their availability and toxicity to algae. Besides metal uptake by algae and complexation of Zn by algal exudates, the rapid decrease in the labile Zn concentration during the algal blooms was ascribed mainly to the adsorption or coprecipitation of Zn ion onto freshly formed iron hydroxides. PMID:10090809

  12. A Solvent-Free Thermosponge Nanoparticle Platform for Efficient Delivery of Labile Proteins

    PubMed Central

    2015-01-01

    Protein therapeutics have gained attention recently for treatment of a myriad of human diseases due to their high potency and unique mechanisms of action. We present the development of a novel polymeric thermosponge nanoparticle for efficient delivery of labile proteins using a solvent-free polymer thermo-expansion mechanism with clinical potential, capable of effectively delivering a range of therapeutic proteins in a sustained manner with no loss of bioactivity, with improved biological half-lives and efficacy in vivo. PMID:25333768

  13. Protein degradation by ubiquitinproteasome system in formation and labilization of contextual conditioning memory

    PubMed Central

    Sol Fustiana, Mara; de la Fuente, Vernica; Federman, Noel; Freudenthal, Ramiro

    2014-01-01

    The ubiquitinproteasome system (UPS) of protein degradation has been evaluated in different forms of neural plasticity and memory. The role of UPS in such processes is controversial. Several results support the idea that the activation of this system in memory consolidation is necessary to overcome negative constrains for plasticity. In this case, the inhibition of the UPS during consolidation impairs memory. Similar results were reported for memory reconsolidation. However, in other cases, the inhibition of UPS had no effect on memory consolidation and reconsolidation but impedes the amnesic action of protein synthesis inhibition after retrieval. The last finding suggests a specific action of the UPS inhibitor on memory labilization. However, another interpretation is possible in terms of the synthesis/degradation balance of positive and negative elements in neural plasticity, as was found in the case of long-term potentiation. To evaluate these alternative interpretations, other reconsolidation-interfering drugs than translation inhibitors should be tested. Here we analyzed initially the UPS inhibitor effect in contextual conditioning in crabs. We found that UPS inhibition during consolidation impaired long-term memory. In contrast, UPS inhibition did not affect memory reconsolidation after contextual retrieval but, in fact, impeded memory labilization, blocking the action of drugs that does not affect directly the protein synthesis. To extend these finding to vertebrates, we performed similar experiments in contextual fear memory in mice. We found that the UPS inhibitor in hippocampus affected memory consolidation and blocked memory labilization after retrieval. These findings exclude alternative interpretations to the requirement of UPS in memory labilization and give evidence of this mechanism in both vertebrates and invertebrates. PMID:25135196

  14. Protein degradation by ubiquitin-proteasome system in formation and labilization of contextual conditioning memory.

    PubMed

    Sol Fustiana, Mara; de la Fuente, Vernica; Federman, Noel; Freudenthal, Ramiro; Romano, Arturo

    2014-09-01

    The ubiquitin-proteasome system (UPS) of protein degradation has been evaluated in different forms of neural plasticity and memory. The role of UPS in such processes is controversial. Several results support the idea that the activation of this system in memory consolidation is necessary to overcome negative constrains for plasticity. In this case, the inhibition of the UPS during consolidation impairs memory. Similar results were reported for memory reconsolidation. However, in other cases, the inhibition of UPS had no effect on memory consolidation and reconsolidation but impedes the amnesic action of protein synthesis inhibition after retrieval. The last finding suggests a specific action of the UPS inhibitor on memory labilization. However, another interpretation is possible in terms of the synthesis/degradation balance of positive and negative elements in neural plasticity, as was found in the case of long-term potentiation. To evaluate these alternative interpretations, other reconsolidation-interfering drugs than translation inhibitors should be tested. Here we analyzed initially the UPS inhibitor effect in contextual conditioning in crabs. We found that UPS inhibition during consolidation impaired long-term memory. In contrast, UPS inhibition did not affect memory reconsolidation after contextual retrieval but, in fact, impeded memory labilization, blocking the action of drugs that does not affect directly the protein synthesis. To extend these finding to vertebrates, we performed similar experiments in contextual fear memory in mice. We found that the UPS inhibitor in hippocampus affected memory consolidation and blocked memory labilization after retrieval. These findings exclude alternative interpretations to the requirement of UPS in memory labilization and give evidence of this mechanism in both vertebrates and invertebrates. PMID:25135196

  15. Substrate lability and plant activity controls greenhouse gas release from Neotropical peatland

    NASA Astrophysics Data System (ADS)

    Sjogersten, Sofie; Hoyos, Jorge; Lomax, Barry; Turner, Ben; Wright, Emma

    2014-05-01

    Almost one third of global CO2 emissions resulting from land use change and substantial CH4 emissions originate from tropical peatlands. However, our understanding of the controls of CO2 and CH4 release from tropical peatlands are limited. The aim of this study was to investigate the role of peat lability and the activity of the vegetation on gas release using a combination of field and laboratory experiments. We demonstrated that peat lability constrained CH4 production to the surface peat under anaerobic conditions. The presence of plants shifted the C balance from a C source to a C sink with respect to CO2 while the activity of the root system strongly influenced CH4 emissions through its impact on soil O2 inputs. Both field and laboratory data suggest a coupling between the photosynthetic activity of the vegetation and the release of both CO2 and CH4 following the circadian rhythm of the dominant plant functional types. Forest clearance for agriculture resulted in elevated CH4 release, which we attribute in part to the cessation of root O2 inputs to the peat. We conclude that high emissions of CO2 and CH4 from forested tropical peatlands are likely driven by labile C inputs from the vegetation but that root O2 release may limit CH4 emissions.

  16. Labile soil carbon inputs mediate the soil microbial community composition and plant residue decomposition rates

    SciTech Connect

    De Graaff, Marie-Anne; Classen, Aimee T; Castro Gonzalez, Hector F; Schadt, Christopher Warren

    2010-01-01

    Root carbon (C) inputs may regulate decomposition rates in soil, and in this study we ask: how do labile C inputs regulate decomposition of plant residues, and soil microbial communities? In a 14 d laboratory incubation, we added C compounds often found in root exudates in seven different concentrations (0, 0.7, 1.4, 3.6, 7.2, 14.4 and 21.7 mg C g{sup -1} soil) to soils amended with and without {sup 13}C-labeled plant residue. We measured CO{sub 2} respiration and shifts in relative fungal and bacterial rRNA gene copy numbers using quantitative polymerase chain reaction (qPCR). Increased labile C input enhanced total C respiration, but only addition of C at low concentrations (0.7 mg C g{sup -1}) stimulated plant residue decomposition (+2%). Intermediate concentrations (1.4, 3.6 mg C g{sup -1}) had no impact on plant residue decomposition, while greater concentrations of C (> 7.2 mg C g{sup -1}) reduced decomposition (-50%). Concurrently, high exudate concentrations (> 3.6 mg C g{sup -1}) increased fungal and bacterial gene copy numbers, whereas low exudate concentrations (< 3.6 mg C g{sup -1}) increased metabolic activity rather than gene copy numbers. These results underscore that labile soil C inputs can regulate decomposition of more recalcitrant soil C by controlling the activity and relative abundance of fungi and bacteria.

  17. Effects of chemical amendments on the lability and speciation of metals in anaerobically digested biosolids.

    PubMed

    Donner, Erica; Brunetti, Gianluca; Zarcinas, Bernie; Harris, Paul; Tavakkoli, Ehsan; Naidu, Ravi; Lombi, Enzo

    2013-10-01

    The interaction of inorganic contaminants present in biosolids with iron, aluminum, and manganese oxy/hydroxides has been advocated as a key mechanism limiting their bioavailability. In this study, we investigated whether this is indeed the case, and further, whether it can be exploited to produce optimized biosolids products through the addition of chemical additives during sewage sludge processing. Experiments were conducted to investigate whether the addition of iron- and aluminum-based amendments (at 5 different rates) during the anaerobic digestion phase of wastewater treatment can effectively change the speciation or lability of contaminant metals (copper, zinc and cadmium) in biosolids destined for use in agriculture. The performance of the bioreactors was monitored throughout and the speciation and lability were determined in both fresh and 3-month aged biosolids using X-ray absorption spectroscopy (Cu, Zn) and isotopic dilution ((65)Cu, (65)Zn, (109)Cd). The tested amendments (FeCl3, Al2(SO4)3, and Al-rich water treatment residual) did not cause significant changes in metal speciation and were of limited use for reducing the lability of contaminant metals in good quality biosolids (suitable for use in agriculture), suggesting that high affinity binding sites were already in excess in these materials. However, the use of chemical amendments may offer advantages in terms of treatment process optimization and may also be beneficial when biosolids are used for contaminated site remediation. PMID:23981056

  18. Life history lability underlies rapid climate niche evolution in the angiosperm clade Montiaceae.

    PubMed

    Matthew Ogburn, R; Edwards, Erika J

    2015-11-01

    Despite the recent focus on phylogenetic niche conservatism in macroevolutionary studies, many clades have diversified widely along multiple niche dimensions. The factors underlying lineage-specific niche lability are still not well understood. We examined morphological and climate niche evolution in Montiaceae (Caryophyllales), an ecologically variable plant lineage distributed primarily along the mountain chains of the western Americas. Montiaceae inhabit a broader range of temperatures than their relatives, with an increase in the evolutionary rate of temperature niche diversification at the node subtending this clade. Within Montiaceae, life history is highly labile and significantly correlated with temperature, with perennials consistently occurring in cooler environments. This elevated evolutionary lability facilitated repeated shifts between habitats as new environments were created by post-Eocene orogenic events and aridification in the western Americas. The shifts between annual and perennial forms are elaborations of an underlying rosette body plan in most cases, and may involve simple alterations in biomass allocation. Montiaceae stand as another clear counterexample to phylogenetic niche conservatism, and demonstrate a mechanism by which pronounced ecological shifts may occur frequently and rapidly among closely related species. PMID:26143714

  19. Comparison of metal lability in air-dried and fresh dewatered drinking water treatment residuals.

    PubMed

    Wang, Changhui; Pei, Yuansheng; Zhao, Yaqian

    2015-01-01

    In this work, the labilities of Al, As, Ba, Be, Ca, Cd, Co, Cr, Cu, Fe, Mg, Mn, Mo, Ni, Pb, Sr, V and Zn in air-dried (for 60 days) and fresh dewatered WTRs were compared using the Toxicity Characteristic Leaching Procedure (TCLP), fractionation, in vitro digestion and a plant enrichment test. The results showed that the air-dried and fresh dewatered WTRs had different properties, e.g., organic matter composition and available nutrients. The air-dried and fresh dewatered WTRs were non-haf zardous according to the TCLP assessment method used in the United States; however, the metals in the two types of WTRs had different lability. Compared with the metals in the fresh dewatered WTRs, those in the air-dried WTRs tended to be in more stable fractions and also exhibited lower bioaccessibility and bioavailability. Therefore, air-drying can decrease the metal lability and thereby reduce the potential metal pollution risk of WTRs. PMID:25560259

  20. Bacillithiol is a major buffer of the labile zinc pool in Bacillus subtilis

    PubMed Central

    Ma, Zhen; Chandrangsu, Pete; Helmann, Tyler C.; Romsang, Adisak; Gaballa, Ahmed; Helmann, John D.

    2014-01-01

    Intracellular zinc levels are tightly regulated since zinc is an essential cofactor for numerous enzymes, yet can be toxic when present in excess. The majority of intracellular zinc is tightly associated with proteins and is incorporated during synthesis from a poorly defined pool of kinetically labile zinc. In Bacillus subtilis, this labile pool is sensed by equilibration with the metalloregulator Zur, as an indication of zinc sufficiency, and by CzrA, as an indication of zinc excess. Here, we demonstrate that the low molecular weight thiol bacillithiol (BSH) serves as a major buffer of the labile zinc pool. Upon shift to conditions of zinc excess, cells transiently accumulate zinc in a low molecular weight pool, and this accumulation is largely dependent on BSH. Cells lacking BSH are more sensitive to zinc stress, and they induce zinc efflux at lower external zinc concentrations. Thiol reactive agents such as diamide and cadmium induce zinc efflux by interfering with the Zn-buffering function of BSH. Our data provide new insights into intracellular zinc buffering and may have broad relevance given the presence of BSH in pathogens and the proposed role of zinc sequestration in innate immunity. PMID:25213752

  1. Botulinum toxin: The Midas touch

    PubMed Central

    Shilpa, P. S.; Kaul, Rachna; Sultana, Nishat; Bhat, Suraksha

    2014-01-01

    Botulinum Toxin (BT) is a natural molecule produced during growth and autolysis of bacterium called Clostridium botulinum. Use of BT for cosmetic purposes has gained popularity over past two decades, and recently, other therapeutic uses of BT has been extensively studied. BT is considered as a minimally invasive agent that can be used in the treatment of various orofacial disorders and improving the quality of life in such patients. The objective of this article is to review the nature, mechanism of action of BT, and its application in various head and neck diseases. PMID:24678189

  2. Designing Inhibitors of Anthrax Toxin

    PubMed Central

    Nestorovich, Ekaterina M.; Bezrukov, Sergey M.

    2014-01-01

    Introduction Present-day rational drug design approaches are based on exploiting unique features of the target biomolecules, small- or macromolecule drug candidates, and physical forces that govern their interactions. The 2013 Nobel Prize in chemistry awarded “for the development of multiscale models for complex chemical systems” once again demonstrated the importance of the tailored drug discovery that reduces the role of the trial and error approach to a minimum. The “rational drug design” term is rather comprehensive as it includes all contemporary methods of drug discovery where serendipity and screening are substituted by the information-guided search for new and existing compounds. Successful implementation of these innovative drug discovery approaches is inevitably preceded by learning the physics, chemistry, and physiology of functioning of biological structures under normal and pathological conditions. Areas covered This article provides an overview of the recent rational drug design approaches to discover inhibitors of anthrax toxin. Some of the examples include small-molecule and peptide-based post-exposure therapeutic agents as well as several polyvalent compounds. The review also directs the reader to the vast literature on the recognized advances and future possibilities in the field. Expert opinion Existing options to combat anthrax toxin lethality are limited. With the only anthrax toxin inhibiting therapy (PA-targeting with a monoclonal antibody, raxibacumab) approved to treat inhalational anthrax, in our view, the situation is still insecure. The FDA’s animal rule for drug approval, which clears compounds without validated efficacy studies on humans, creates a high level of uncertainty, especially when a well-characterized animal model does not exist. Besides, unlike PA, which is known to be unstable, LF remains active in cells and in animal tissues for days. Therefore, the effectiveness of the post-exposure treatment of the individuals with anti-PA therapeutics can be time-dependent, requiring coordinated use of membrane permeable small-molecule inhibitors, which block the LF and EF enzymatic activity intracellularly. The desperate search for an ideal anthrax antitoxin allowed researchers to gain important knowledge of the basic principles of small-molecule interactions with their protein targets that could be easily transferred to other systems. At the same time, better identification and validation of anthrax toxin therapeutic targets at the molecular level, which include understanding of the physical forces underlying the target/drug interaction, as well as elucidation of the parameters determining the corresponding therapeutic windows, require further examination. PMID:24447197

  3. Biologic response to environmental toxins

    SciTech Connect

    Lerner, S.

    1983-12-01

    Biological response to environmental toxins results from the sum of natural, environmental, avocational, inapparent, and occupational exposures. These external exposures result in acceptable or unacceptable levels of absorption or internal exposure based on anticipated biological effects. There is no level of exposure which is in and of itself synonymous with intoxication. Biological effects may be classified as physiologic or pathologic, adaptive or nonadaptive, respectively. In each instance, the response may be acceptable or unacceptable. Intoxication requires the demonstration of a significant impairment of health. One may have an unacceptable pathologic response and still not have intoxication. Professional judgment is required.

  4. Assessing the Selectivity of Extractant Solutions for Recovering Labile Arsenic Associated with Iron (Hydr)oxides and Sulfides in Sediments

    EPA Science Inventory

    Sequential extractions can provide analytical constraints on the identification of mineral phases that control arsenic speciation in sediments. Model solids were used in this study to evaluate different solutions designed to extract arsenic from relatively labile solid phases. ...

  5. Plant Insecticidal Toxins in Ecological Networks

    PubMed Central

    Ibanez, Sébastien; Gallet, Christiane; Després, Laurence

    2012-01-01

    Plant secondary metabolites play a key role in plant-insect interactions, whether constitutive or induced, C- or N-based. Anti-herbivore defences against insects can act as repellents, deterrents, growth inhibitors or cause direct mortality. In turn, insects have evolved a variety of strategies to act against plant toxins, e.g., avoidance, excretion, sequestration and degradation of the toxin, eventually leading to a co-evolutionary arms race between insects and plants and to co-diversification. Anti-herbivore defences also negatively impact mutualistic partners, possibly leading to an ecological cost of toxin production. However, in other cases toxins can also be used by plants involved in mutualistic interactions to exclude inadequate partners and to modify the cost/benefit ratio of mutualism to their advantage. When considering the whole community, toxins have an effect at many trophic levels. Aposematic insects sequester toxins to defend themselves against predators. Depending on the ecological context, toxins can either increase insects’ vulnerability to parasitoids and entomopathogens or protect them, eventually leading to self-medication. We conclude that studying the community-level impacts of plant toxins can provide new insights into the synthesis between community and evolutionary ecology. PMID:22606374

  6. Stool Test: C. Difficile Toxin (For Parents)

    MedlinePLUS

    ... Your Child All About Food Allergies Stool Test: C. Difficile Toxin KidsHealth > For Parents > Stool Test: C. Difficile Toxin Print A A A Text Size ... Questions en español Muestra de materia fecal: toxina C. difficile What It Is A stool (feces) sample ...

  7. Botulinum Toxin and Gastrointestinal Tract Disorders

    PubMed Central

    Weiser, Kirsten; Kennedy, Abigail

    2008-01-01

    The history of botulinum toxin is fascinating. First recognized as the cause of botulism nearly 200 years ago, it was originally feared as a deadly poison. Over the last 30 years, however, botulinum toxin has been transformed into a readily available medication used to treat a variety of medical disorders. Interest in the use of botulinum toxin has been particularly strong for patients with spastic smooth muscle disorders of the gastrointestinal tract. Patients with achalasia, diffuse esophageal spasm, gastroparesis, sphincter of Oddi dysfunction, and anal fissures have all been treated with botulinum toxin injections, often with impressive results. However, not all patients respond to botulinum toxin therapy, and large randomized controlled trials are lacking for many conditions commonly treated with botulinum toxin. This paper reviews the history, microbiology, and pharmacology of botulinum toxin, discusses its mechanism of action, and then presents recent evidence from the literature regarding the use of botulinum toxin for the treatment of a variety of gastrointestinal tract disorders. PMID:21960915

  8. Target-Driven Evolution of Scorpion Toxins

    PubMed Central

    Zhang, Shangfei; Gao, Bin; Zhu, Shunyi

    2015-01-01

    It is long known that peptide neurotoxins derived from a diversity of venomous animals evolve by positive selection following gene duplication, yet a force that drives their adaptive evolution remains a mystery. By using maximum-likelihood models of codon substitution, we analyzed molecular adaptation in scorpion sodium channel toxins from a specific species and found ten positively selected sites, six of which are located at the core-domain of scorpion ?-toxins, a region known to interact with two adjacent loops in the voltage-sensor domain (DIV) of sodium channels, as validated by our newly constructed computational model of toxin-channel complex. Despite the lack of positive selection signals in these two loops, they accumulated extensive sequence variations by relaxed purifying selection in prey and predators of scorpions. The evolutionary variability in the toxin-bound regions of sodium channels indicates that accelerated substitutions in the multigene family of scorpion toxins is a consequence of dealing with the target diversity. This work presents an example of atypical co-evolution between animal toxins and their molecular targets, in which toxins suffered from more prominent selective pressure from the channels of their competitors. Our discovery helps explain the evolutionary rationality of gene duplication of toxins in a specific venomous species. PMID:26444071

  9. Target-Driven Evolution of Scorpion Toxins.

    PubMed

    Zhang, Shangfei; Gao, Bin; Zhu, Shunyi

    2015-01-01

    It is long known that peptide neurotoxins derived from a diversity of venomous animals evolve by positive selection following gene duplication, yet a force that drives their adaptive evolution remains a mystery. By using maximum-likelihood models of codon substitution, we analyzed molecular adaptation in scorpion sodium channel toxins from a specific species and found ten positively selected sites, six of which are located at the core-domain of scorpion α-toxins, a region known to interact with two adjacent loops in the voltage-sensor domain (DIV) of sodium channels, as validated by our newly constructed computational model of toxin-channel complex. Despite the lack of positive selection signals in these two loops, they accumulated extensive sequence variations by relaxed purifying selection in prey and predators of scorpions. The evolutionary variability in the toxin-bound regions of sodium channels indicates that accelerated substitutions in the multigene family of scorpion toxins is a consequence of dealing with the target diversity. This work presents an example of atypical co-evolution between animal toxins and their molecular targets, in which toxins suffered from more prominent selective pressure from the channels of their competitors. Our discovery helps explain the evolutionary rationality of gene duplication of toxins in a specific venomous species. PMID:26444071

  10. Formation and Control of Cyanobacterial Toxins

    EPA Science Inventory

    This presentation will cover the formation of harmful algal blooms and the control of their toxins. Data will be presented from current ORD projects on the treatment of cyanobacterial toxins through drinking water treatment facilities. The results will demonstrate that current c...

  11. Cholesteryl ester hydroperoxide lability is a key feature of the oxidative susceptibility of small, dense LDL.

    PubMed

    Chancharme, L; Thérond, P; Nigon, F; Lepage, S; Couturier, M; Chapman, M J

    1999-03-01

    Abundant evidence has been provided to substantiate the elevated cardiovascular risk associated with small, dense, low density lipoprotein (LDL) particles. The diminished resistance of dense LDL to oxidative stress in both normolipidemic and dyslipidemic subjects is established; nonetheless, the molecular basis of this phenomenon remains indeterminate. We have defined the primary molecular targets of lipid hydroperoxide formation in light, intermediate, and dense subclasses of LDL after copper-mediated oxidation and have compared the relative stabilities of the hydroperoxide derivatives of phospholipids and cholesteryl esters (CEs) as a function of the time course of oxidation. LDL subclasses (LDL1 through LDL5) were isolated from normolipidemic plasma by isopycnic density gradient ultracentrifugation, and their content of polyunsaturated molecular species of phosphatidylcholine (PC) and CE and of lipophilic antioxidants was quantified by reverse-phase high-performance liquid chromatography. The molar ratio of the particle content of polyunsaturated CE and PC species containing linoleate or arachidonate relative to alpha-tocopherol or beta-carotene did not differ significantly between LDL subspecies. Nonetheless, dense LDL contained significantly less polyunsaturated CE species (400 mol per particle) compared with LDL1 through LDL4 (range, approximately 680 to 490 mol per particle). Although the formation of PC-derived hydroperoxides did not vary significantly between LDL subspecies as a function of the time course of copper-mediated oxidation, the abundance of the C18:2 and C20:4 CE hydroperoxides was uniquely deficient in dense LDL (23 and 0.6 mol per particle, respectively, in LDL5; 47 to 58 and 1.9 to 2.3 mol per particle, respectively, in other LDL subclasses) at propagation half-time. When expressed as a lability ratio (mol hydroperoxides formed relative to each 100 mol of substrate consumed) at half-time, the oxidative lability of CE hydroperoxides in dense LDL was significantly elevated (lability ratio <25:100) relative to that in lighter, larger LDL particle subclasses (lability ratio >40:100) throughout the oxidative time course. We conclude that the elevated lability of CE hydroperoxides in dense LDL underlies the diminished oxidative resistance of these particles. Moreover, this phenomenon appears to result not only from the significantly elevated PC to free cholesterol ratio (1.54:1) in dense LDL particles (1.15:1 to 1.25:1 for other LDL subclasses) but also from their unique structural features, including a distinct apoB100 conformation, which may facilitate covalent bond formation between oxidized CE and apoB100. PMID:10073990

  12. Molecular insights into the microbial formation of marine dissolved organic matter: recalcitrant or labile?

    NASA Astrophysics Data System (ADS)

    Koch, B. P.; Kattner, G.; Witt, M.; Passow, U.

    2014-08-01

    The degradation of marine dissolved organic matter (DOM) is an important control variable in the global carbon cycle. For our understanding of the kinetics of organic matter cycling in the ocean, it is crucial to achieve a mechanistic and molecular understanding of its transformation processes. A long-term microbial experiment was performed to follow the production of non-labile DOM by marine bacteria. Two different glucose concentrations and dissolved algal exudates were used as substrates. We monitored the bacterial abundance, concentrations of dissolved and particulate organic carbon (DOC, POC), nutrients, amino acids and transparent exopolymer particles (TEP) for 2 years. The molecular characterization of extracted DOM was performed by ultrahigh resolution Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) after 70 days and after ∼2 years of incubation. Although glucose quickly degraded, a non-labile DOC background (5-9% of the initial DOC) was generated in the glucose incubations. Only 20% of the organic carbon from the algal exudate degraded within the 2 years of incubation. The degradation rates for the non-labile DOC background in the different treatments varied between 1 and 11 ?mol DOC L-1 year-1. Transparent exopolymer particles, which are released by microorganisms, were produced during glucose degradation but decreased back to half of the maximum concentration within less than 3 weeks (degradation rate: 25 ?g xanthan gum equivalents L-1 d-1) and were below detection in all treatments after 2 years. Additional glucose was added after 2 years to test whether labile substrate can promote the degradation of background DOC (co-metabolism; priming effect). A priming effect was not observed but the glucose addition led to a slight increase of background DOC. The molecular analysis demonstrated that DOM generated during glucose degradation differed appreciably from DOM transformed during the degradation of the algal exudates. Our results led to several conclusions: (i) based on our experimental setup, higher substrate concentration resulted in a higher concentration of non-labile DOC; (ii) TEP, generated by bacteria, degrade rapidly, thus limiting their potential contribution to carbon sequestration; (iii) the molecular signatures of DOM derived from algal exudates and glucose after 70 days of incubation differed strongly from refractory DOM. After 2 years, however, the molecular patterns of DOM in glucose incubations were more similar to deep ocean DOM whereas the degraded exudate was still different.

  13. The evolution of labile traits in sex- and age-structured populations.

    PubMed

    Childs, Dylan Z; Sheldon, Ben C; Rees, Mark

    2016-03-01

    Many quantitative traits are labile (e.g. somatic growth rate, reproductive timing and investment), varying over the life cycle as a result of behavioural adaptation, developmental processes and plastic responses to the environment. At the population level, selection can alter the distribution of such traits across age classes and among generations. Despite a growing body of theoretical research exploring the evolutionary dynamics of labile traits, a data-driven framework for incorporating such traits into demographic models has not yet been developed. Integral projection models (IPMs) are increasingly being used to understand the interplay between changes in labile characters, life histories and population dynamics. One limitation of the IPM approach is that it relies on phenotypic associations between parents and offspring traits to capture inheritance. However, it is well-established that many different processes may drive these associations, and currently, no clear consensus has emerged on how to model micro-evolutionary dynamics in an IPM framework. We show how to embed quantitative genetic models of inheritance of labile traits into age-structured, two-sex models that resemble standard IPMs. Commonly used statistical tools such as GLMs and their mixed model counterparts can then be used for model parameterization. We illustrate the methodology through development of a simple model of egg-laying date evolution, parameterized using data from a population of Great tits (Parus major). We demonstrate how our framework can be used to project the joint dynamics of species' traits and population density. We then develop a simple extension of the age-structured Price equation (ASPE) for two-sex populations, and apply this to examine the age-specific contributions of different processes to change in the mean phenotype and breeding value. The data-driven framework we outline here has the potential to facilitate greater insight into the nature of selection and its consequences in settings where focal traits vary over the lifetime through ontogeny, behavioural adaptation and phenotypic plasticity, as well as providing a potential bridge between theoretical and empirical studies of labile trait variation. PMID:26899421

  14. Brown spider dermonecrotic toxin directly induces nephrotoxicity

    SciTech Connect

    Chaim, Olga Meiri; Sade, Youssef Bacila; Bertoni da Silveira, Rafael; Toma, Leny; Kalapothakis, Evanguedes; Chavez-Olortegui, Carlos; Mangili, Oldemir Carlos; Gremski, Waldemiro; Dietrich, Carl Peter von; Nader, Helena B.; Sanches Veiga, Silvio . E-mail: veigass@ufpr.br

    2006-02-15

    Brown spider (Loxosceles genus) venom can induce dermonecrotic lesions at the bite site and systemic manifestations including fever, vomiting, convulsions, disseminated intravascular coagulation, hemolytic anemia and acute renal failure. The venom is composed of a mixture of proteins with several molecules biochemically and biologically well characterized. The mechanism by which the venom induces renal damage is unknown. By using mice exposed to Loxosceles intermedia recombinant dermonecrotic toxin (LiRecDT), we showed direct induction of renal injuries. Microscopic analysis of renal biopsies from dermonecrotic toxin-treated mice showed histological alterations including glomerular edema and tubular necrosis. Hyalinization of tubules with deposition of proteinaceous material in the tubule lumen, tubule epithelial cell vacuoles, tubular edema and epithelial cell lysis was also observed. Leukocytic infiltration was neither observed in the glomerulus nor the tubules. Renal vessels showed no sign of inflammatory response. Additionally, biochemical analyses showed such toxin-induced changes in renal function as urine alkalinization, hematuria and azotemia with elevation of blood urea nitrogen levels. Immunofluorescence with dermonecrotic toxin antibodies and confocal microscopy analysis showed deposition and direct binding of this toxin to renal intrinsic structures. By immunoblotting with a hyperimmune dermonecrotic toxin antiserum on renal lysates from toxin-treated mice, we detected a positive signal at the region of 33-35 kDa, which strengthens the idea that renal failure is directly induced by dermonecrotic toxin. Immunofluorescence reaction with dermonecrotic toxin antibodies revealed deposition and binding of this toxin directly in MDCK epithelial cells in culture. Similarly, dermonecrotic toxin treatment caused morphological alterations of MDCK cells including cytoplasmic vacuoles, blebs, evoked impaired spreading and detached cells from each other and from culture substratum. In addition, dermonecrotic toxin treatment of MDCK cells changed their viability evaluated by XTT and Neutral-Red Uptake methodologies. The present results point to brown spider dermonecrotic toxin cytotoxicity upon renal structures in vivo and renal cells in vitro and provide experimental evidence that this brown spider toxin is directly involved in nephrotoxicity evoked during Loxosceles spider venom accidents.

  15. The toxin component of targeted anti-tumor toxins determines their efficacy increase by saponins.

    PubMed

    Weng, Alexander; Thakur, Mayank; Beceren-Braun, Figen; Bachran, Diana; Bachran, Christopher; Riese, Sebastian B; Jenett-Siems, Kristina; Gilabert-Oriol, Roger; Melzig, Matthias F; Fuchs, Hendrik

    2012-06-01

    Tumor-targeting protein toxins are composed of a toxic enzyme coupled to a specific cell binding domain that targets cancer-associated antigens. The anti-tumor treatment by targeted toxins is accompanied by dose-limiting side effects. The future prospects of targeted toxins for therapeutic use in humans will be determined by reduce side effects. Certain plant secondary metabolites (saponins) were shown to increase the efficacy of a particular epidermal growth factor receptor (EGFR)-targeted toxin, paralleled by a tremendous decrease of side effects. This study was conducted in order to investigate the effects of substituting different toxin moieties fused to an EGF ligand binding domain on the augmentative ability of saponins for each against therapeutic potential of the saponin-mediated efficacy increase for different anti-tumor toxins targeting the EGFR. We designed several EGFR-targeted toxins varying in the toxic moiety. Each targeted toxin was used in combination with a purified saponin (SA1641), isolated from the ornamental plant Gypsophila paniculata L. SA1641 was characterized and the SA1641-mediated efficacy increase was investigated on EGFR-transfected NIH-3T3 cells. We observed a high dependency of the SA1641-mediated efficacy increase on the nature of toxin used for the construction of the targeted toxin, indicating high specificity. Structural alignments revealed a high homology between saporin and dianthin-30, the two toxic moieties that benefit most from the combination with SA1641. We further demonstrate that SA1641 did not influence the plasma membrane permeability, indicating an intracellular interaction of SA1641 and the toxin components of targeted toxins. Surface plasmon resonance measurements point to a transient binding of SA1641 to the toxin components of targeted toxins. PMID:22309811

  16. Botulinum Toxin in Poststroke Spasticity

    PubMed Central

    Ozcakir, Suheda; Sivrioglu, Koncuy

    2007-01-01

    Poststroke hemiparesis, together with abnormal muscle tone, is a major cause of morbidity and disability. Although most hemiparetic patients are able to reach different ambulatory levels with rehabilitation efforts, upper and lower limb spasticity can impede activities of daily living, personal hygiene, ambulation and, in some cases, functional improvement. The goals of spasticity management include increasing mobility and range of motion, attaining better hygiene, improving splint wear and other functional activities. Conservative measures, such as positioning, stretching and exercise are essential in spasticity management, but alone often are inadequate to effectively control it. Oral antispastic medications often provide limited effects with short duration and frequent unwanted systemic side effects, such as weakness, sedation and dry mouth. Therefore, neuromuscular blockade by local injections have become the first choice for the treatment of focal spasticity, particularly in stroke patients. Botulinum toxin (BTX), being one of the most potent biological toxins, acts by blocking neuromuscular transmission via inhibiting acetylcholine release. Currently, focal spasticity is being treated successfully with BTX via injecting in the spastic muscles. Two antigenically distinct serotypes of BTX are available on the market as type A and B. Clinical studies of BTX used for spastic hemiplegic patients are reviewed in this article in two major categories, upper and lower limb applications. This review addresses efficacy in terms of outcome measures, such as muscle tone reduction and functional outcome, as well as safety issues. Application modifications of dose, dilutions, site of injections and combination therapies with BTX injections are also discussed. PMID:17607049

  17. Biological methods for marine toxin detection.

    PubMed

    Vilario, Natalia; Louzao, M Carmen; Vieytes, Mercedes R; Botana, Luis M

    2010-07-01

    The presence of marine toxins in seafood poses a health risk to human consumers which has prompted the regulation of the maximum content of marine toxins in seafood in the legislations of many countries. Most marine toxin groups are detected by animal bioassays worldwide. Although this method has well known ethical and technical drawbacks, it is the official detection method for all regulated phycotoxins except domoic acid. Much effort by the scientific and regulatory communities has been focused on the development of alternative techniques that enable the substitution or reduction of bioassays; some of these have recently been included in the official detection method list. During the last two decades several biological methods including use of biosensors have been adapted for detection of marine toxins. The main advances in marine toxin detection using this kind of technique are reviewed. Biological methods offer interesting possibilities for reduction of the number of biosassays and a very promising future of new developments. PMID:20458470

  18. The Enterotoxicity of Clostridium difficile Toxins

    PubMed Central

    Sun, Xingmin; Savidge, Tor; Feng, Hanping

    2010-01-01

    The major virulence factors of Clostridium difficile infection (CDI) are two large exotoxins A (TcdA) and B (TcdB). However, our understanding of the specific roles of these toxins in CDI is still evolving. It is now accepted that both toxins are enterotoxic and proinflammatory in the human intestine. Both purified TcdA and TcdB are capable of inducing the pathophysiology of CDI, although most studies have focused on TcdA. C. difficile toxins exert a wide array of biological activities by acting directly on intestinal epithelial cells. Alternatively, the toxins may target immune cells and neurons once the intestinal epithelial barrier is disrupted. The toxins may also act indirectly by stimulating cells to produce chemokines, proinflammatory cytokines, neuropeptides and other neuroimmune signals. This review considers the mechanisms of TcdA- and TcdB-induced enterotoxicity, and recent developments in this field. PMID:22069662

  19. Crystallization of isoelectrically homogeneous cholera toxin

    SciTech Connect

    Spangler, B.D.; Westbrook, E.M. )

    1989-02-07

    Past difficulty in growing good crystals of cholera toxin has prevented the study of the crystal structure of this important protein. The authors have determined that failure of cholera toxin to crystallize well has been due to its heterogeneity. They have now succeeded in overcoming the problem by isolating a single isoelectric variant of this oligomeric protein (one A subunit and five B subunits). Cholera toxin purified by their procedure readily forms large single crystals. The crystal form has been described previously. They have recorded data from native crystals of cholera toxin to 3.0-{angstrom} resolution with our electronic area detectors. With these data, they have found the orientation of a 5-fold symmetry axis within these crystals, perpendicular to the screw dyad of the crystal. They are now determining the crystal structure of cholera toxin by a combination of multiple heavy-atom isomorphous replacement and density modification techniques, making use of rotational 5-fold averaging of the B subunits.

  20. Toxins and Secretion Systems of Photorhabdus luminescens

    PubMed Central

    Rodou, Athina; Ankrah, Dennis O.; Stathopoulos, Christos

    2010-01-01

    Photorhabdus luminescens is a nematode-symbiotic, gram negative, bioluminescent bacterium, belonging to the family of Enterobacteriaceae. Recent studies show the importance of this bacterium as an alternative source of insecticides, as well as an emerging human pathogen. Various toxins have been identified and characterized in this bacterium. These toxins are classified into four major groups: the toxin complexes (Tcs), the Photorhabdus insect related (Pir) proteins, the makes caterpillars floppy (Mcf) toxins and the Photorhabdus virulence cassettes (PVC); the mechanisms however of toxin secretion are not fully elucidated. Using bioinformatics analysis and comparison against the components of known secretion systems, multiple copies of components of all known secretion systems, except the ones composing a type IV secretion system, were identified throughout the entire genome of the bacterium. This indicates that Photorhabdus luminescens has all the necessary means for the secretion of virulence factors, thus it is capable of establishing a microbial infection. PMID:22069636

  1. Evaluation of synergism among Bacillus thuringiensis toxins.

    PubMed Central

    Tabashnik, B E

    1992-01-01

    A simple test for synergism among toxins is described and applied to previously reported data on independent and joint toxicities of insecticidal proteins from Bacillus thuringiensis. The analysis shows synergism between a 27-kDa (CytA) toxin and 130- or 65-kDa (CryIV) toxins from B. thuringiensis subsp. israelensis against Aedes aegypti larvae. No positive synergism between 130- and 65-kDa toxins or among three CryIA toxins tested against seven species of Lepidoptera occurred. Comparisons with the original interpretations of these data show one case in which synergism occurred but was reported previously as absent and two cases that were not synergistic but were reported previously as suggestive of synergism. These results show that lack of an appropriate test for synergism can produce misleading conclusions. The methods described here can be used to test for synergistic effects of any poisons. PMID:1444368

  2. Toxins

    MedlinePLUS

    ... trace metals and others. In: Goldman L, Schafer AI, eds. Goldman-Cecil Medicine . 25th ed. Philadelphia, PA: ... Ford MD. Acute poisoning. In: Goldman L, Schafer AI, eds. Goldman-Cecil Medicine . 25th ed. Philadelphia, PA: ...

  3. Roles of Anthrax Toxin Receptor 2 in Anthrax Toxin Membrane Insertion and Pore Formation

    PubMed Central

    Sun, Jianjun; Jacquez, Pedro

    2016-01-01

    Interaction between bacterial toxins and cellular surface receptors is an important component of the host-pathogen interaction. Anthrax toxin protective antigen (PA) binds to the cell surface receptor, enters the cell through receptor-mediated endocytosis, and forms a pore on the endosomal membrane that translocates toxin enzymes into the cytosol of the host cell. As the major receptor for anthrax toxin in vivo, anthrax toxin receptor 2 (ANTXR2) plays an essential role in anthrax toxin action by providing the toxin with a high-affinity binding anchor on the cell membrane and a path of entry into the host cell. ANTXR2 also acts as a molecular clamp by shifting the pH threshold of PA pore formation to a more acidic pH range, which prevents premature pore formation at neutral pH before the toxin reaches the designated intracellular location. Most recent studies have suggested that the disulfide bond in the immunoglobulin (Ig)-like domain of ANTXR2 plays an essential role in anthrax toxin action. Here we will review the roles of ANTXR2 in anthrax toxin action, with an emphasis on newly updated knowledge. PMID:26805886

  4. Roles of Anthrax Toxin Receptor 2 in Anthrax Toxin Membrane Insertion and Pore Formation.

    PubMed

    Sun, Jianjun; Jacquez, Pedro

    2016-01-01

    Interaction between bacterial toxins and cellular surface receptors is an important component of the host-pathogen interaction. Anthrax toxin protective antigen (PA) binds to the cell surface receptor, enters the cell through receptor-mediated endocytosis, and forms a pore on the endosomal membrane that translocates toxin enzymes into the cytosol of the host cell. As the major receptor for anthrax toxin in vivo, anthrax toxin receptor 2 (ANTXR2) plays an essential role in anthrax toxin action by providing the toxin with a high-affinity binding anchor on the cell membrane and a path of entry into the host cell. ANTXR2 also acts as a molecular clamp by shifting the pH threshold of PA pore formation to a more acidic pH range, which prevents premature pore formation at neutral pH before the toxin reaches the designated intracellular location. Most recent studies have suggested that the disulfide bond in the immunoglobulin (Ig)-like domain of ANTXR2 plays an essential role in anthrax toxin action. Here we will review the roles of ANTXR2 in anthrax toxin action, with an emphasis on newly updated knowledge. PMID:26805886

  5. Activation of syndecan-1 ectodomain shedding by Staphylococcus aureus alpha-toxin and beta-toxin.

    TOXLINE Toxicology Bibliographic Information

    Park PW; Foster TJ; Nishi E; Duncan SJ; Klagsbrun M; Chen Y

    2004-01-02

    Exploitation of host components by microbes to promote their survival in the hostile host environment has been a recurring theme in recent years. Available data indicate that bacterial pathogens activate ectodomain shedding of host cell surface molecules to enhance their virulence. We reported previously that several major bacterial pathogens activate ectodomain shedding of syndecan-1, the major heparan sulfate proteoglycan of epithelial cells. Here we define the molecular basis of how Staphylococcus aureus activates syndecan-1 shedding. We screened mutant S. aureus strains devoid of various toxin and protease genes and found that only strains lacking both alpha-toxin and beta-toxin genes do not stimulate shedding. Mutations in the agr global regulatory locus, which positively regulates expression of alpha- and beta-toxins and other exoproteins, also abrogated the capacity to stimulate syndecan-1 shedding. Furthermore, purified S. aureus alpha- and beta-toxins, but not enterotoxin A and toxic shock syndrome toxin-1, rapidly potentiated shedding in a concentration-dependent manner. These results establish that S. aureus activates syndecan-1 ectodomain shedding via its two virulence factors, alpha- and beta-toxins. Toxin-activated shedding was also selectively inhibited by antagonists of the host cell shedding mechanism, indicating that alpha- and beta-toxins shed syndecan-1 ectodomains through stimulation of the host cell's shedding machinery. Interestingly, beta-toxin, but not alpha-toxin, also enhanced ectodomain shedding of syndecan-4 and heparin-binding epidermal growth factor. Because shedding of these ectodomains has been implicated in promoting bacterial pathogenesis, activation of ectodomain shedding by alpha-toxin and beta-toxin may be a previously unknown virulence mechanism of S. aureus.

  6. Interplay between Toxin Transport and Flotillin Localization

    PubMed Central

    Pust, Sascha; Dyve, Anne Berit; Torgersen, Maria L.; van Deurs, Bo; Sandvig, Kirsten

    2010-01-01

    The flotillin proteins are localized in lipid domains at the plasma membrane as well as in intracellular compartments. In the present study, we examined the importance of flotillin-1 and flotillin-2 for the uptake and transport of the bacterial Shiga toxin (Stx) and the plant toxin ricin and we investigated whether toxin binding and uptake were associated with flotillin relocalization. We observed a toxin-induced redistribution of the flotillins, which seemed to be regulated in a p38-dependent manner. Our experiments provide no evidence for a changed endocytic uptake of Stx or ricin in cells silenced for flotillin-1 or -2. However, the Golgi-dependent sulfation of both toxins was significantly reduced in flotillin knockdown cells. Interestingly, when the transport of ricin to the ER was investigated, we obtained an increased mannosylation of ricin in flotillin-1 and flotillin-2 knockdown cells. The toxicity of both toxins was twofold increased in flotillin-depleted cells. Since BFA (Brefeldin A) inhibits the toxicity even in flotillin knockdown cells, the retrograde toxin transport is apparently still Golgi-dependent. Thus, flotillin proteins regulate and facilitate the retrograde transport of Stx and ricin. PMID:20107503

  7. Fungi and fungal toxins as weapons.

    PubMed

    Paterson, R Russell M

    2006-09-01

    Recent aggressive attacks on innocent citizens have resulted in governments increasing security. However, there is a good case for prevention rather than reaction. Bioweapons, mycotoxins, fungal biocontrol agents (FBCA), and even pharmaceuticals contain, or are, toxins and need to be considered in the context of the new paradigm. Is it desirable to discuss such issues? None of the fungi are (a) as toxic as botulinum toxin from Clostridium botulinum, and (b) as dangerous as nuclear weapons. One toxin may be defined as a pharmaceutical and vice versa simply by a small change in concentration or a moiety. Mycotoxins are defined as naturally occurring toxic compounds obtained from fungi. They are the biggest chronic health risk when incorporated into the diet. The current list of fungal toxins as biochemical weapons is small, although awareness is growing of the threats they may pose. T-2 toxin is perhaps the biggest concern. A clear distinction is required between the biological (fungus) and chemical (toxin) aspects of the issue. There is an obvious requirement to be able to trace these fungi and compounds in the environment and to know when concentrations are abnormal. Many FBCA, produce toxins. This paper indicates how to treat mycotoxicosis and decontaminate mycotoxins. There is considerable confusion and inconsistency surrounding this topic which requires assessment in an impartial and scientific manner. PMID:16908123

  8. Clostridium difficile: its disease and toxins.

    PubMed Central

    Lyerly, D M; Krivan, H C; Wilkins, T D

    1988-01-01

    Clostridium difficile is the etiologic agent of pseudomembranous colitis, a severe, sometimes fatal disease that occurs in adults undergoing antimicrobial therapy. The disease, ironically, has been most effectively treated with antibiotics, although some of the newer methods of treatment such as the replacement of the bowel flora may prove more beneficial for patients who continue to relapse with pseudomembranous colitis. The organism produces two potent exotoxins designated toxin A and toxin B. Toxin A is an enterotoxin believed to be responsible for the diarrhea and mucosal tissue damage which occur during the disease. Toxin B is an extremely potent cytotoxin, but its role in the disease has not been as well studied. There appears to be a cascade of events which result in the expression of the activity of these toxins, and these events, ranging from the recognition of a trisaccharide receptor by toxin A to the synergistic action of the toxins and their possible dissemination in the body, are discussed in this review. The advantages and disadvantages of the various assays, including tissue culture assay, enzyme immunoassay, and latex agglutination, currently used in the clinical diagnosis of the disease also are discussed. PMID:3144429

  9. Cyanobacterial toxins: risk management for health protection

    SciTech Connect

    Codd, Geoffrey A.; Morrison, Louise F.; Metcalf, James S

    2005-03-15

    This paper reviews the occurrence and properties of cyanobacterial toxins, with reference to the recognition and management of the human health risks which they may present. Mass populations of toxin-producing cyanobacteria in natural and controlled waterbodies include blooms and scums of planktonic species, and mats and biofilms of benthic species. Toxic cyanobacterial populations have been reported in freshwaters in over 45 countries, and in numerous brackish, coastal, and marine environments. The principal toxigenic genera are listed. Known sources of the families of cyanobacterial toxins (hepato-, neuro-, and cytotoxins, irritants, and gastrointestinal toxins) are briefly discussed. Key procedures in the risk management of cyanobacterial toxins and cells are reviewed, including derivations (where sufficient data are available) of tolerable daily intakes (TDIs) and guideline values (GVs) with reference to the toxins in drinking water, and guideline levels for toxigenic cyanobacteria in bathing waters. Uncertainties and some gaps in knowledge are also discussed, including the importance of exposure media (animal and plant foods), in addition to potable and recreational waters. Finally, we present an outline of steps to develop and implement risk management strategies for cyanobacterial cells and toxins in waterbodies, with recent applications and the integration of Hazard Assessment Critical Control Point (HACCP) principles.

  10. MATERIALS FOR BINDING MYCOTOXINS AND THEIR USE IN TOXIN DETECTION

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Materials capable of binding mycotoxins find numerous uses. These include components of test kits for toxin detection, as agents for isolation of toxins from foods or toxin purification, and as binding agents to prevent toxin uptake and thereby protect domestic animals. There is no shortage of mat...

  11. T-2 toxin degradation by micromycetes.

    PubMed

    Jesensk, Z; Sajbidorov, I

    1991-01-01

    The biodegradation of T-2 toxin was studied by strains of micromycetes which were isolated from the environment. The 26 tested strains were divided into three groups. Group contains strains which degraded T-2 toxin very fast. This toxin could not be chromatographically determined in the medium even after 48 hours of incubation and the antifungal activity of residua against Kluyveromyces fragilis CCY-51-1-2 was low or zero. There were strains of Alternaria sp., Ulocladium sp., Aspergillus candidus, Cladosporium cladosporioides, Rhodotorula sp., Aspergillus flavus and Cladosporium macrocarpum. Group II contains with a low activity and in group III the results were variable and non stable. PMID:1880407

  12. Botulinum toxin and intractable trigeminal neuralgia.

    PubMed

    Trk, Ulk; Ilhan, Selen; Alp, Recep; Sur, Haydar

    2005-01-01

    The effects of botulinum toxin have been demonstrated in pain syndromes such as migraine, tension headache, and postherpetic neuralgia. With this background data in hand, the authors planned a randomized, open-ended study to investigate the efficacy of botulinum injections in cases of refractory trigeminal neuralgia. In 8 patients with trigeminal neuralgia, 100 U botulinum toxin was injected into the region of the zygomatic arch. The results of their statistical analyses demonstrated that the medication can be effective in treating trigeminal neuralgia. The patients did not develop any significant adverse effects. Botulinum toxin can be used in the treatment of refractory trigeminal neuralgia. PMID:16062093

  13. Total and Labile Phosphorus Concentrations as Influenced by Riparian Buffer Soil Properties.

    PubMed

    Young, Eric O; Ross, Donald S

    2016-01-01

    Riparian buffers can act as a phosphorus (P) source under active stream bank erosion. Using soil and landscape variables (soil series, drainage class, organic matter, and pH) to index P concentrations could improve P loss risk tools for buffers. The objectives of this study were (i) to determine if soil properties could predict total and labile P concentrations within a 10-ha riparian buffer and (ii) to quantify the degree of spatial dependence of P and related properties. Soil samples were taken in 15-cm increments to a depth of 60 cm using a grid ( = 71) from an established riparian buffer along the Rock River in Vermont. Total soil P (TP), plant-available P determined by Modified Morgan extraction (MM-P), pH, soil organic matter (SOM), soil texture, and select cations were measured. We found that TP (152-1536 mg P kg) and MM-P (0.4-14.6 mg kg) ranged widely, with distinct differences between soil series. Mean TP and MM-P were greater in alluvial and glaciolacustrine soils compared with glacial till. Across all samples, MM-P was weakly related to soil properties; however, total labile P (orthophosphate + organic P measured by ICP) and unreactive labile P (ICP-P - colorimetric-P) could both be predicted by SOM ( = 0.59 and 0.73, respectively). Strong spatial dependence was found for P and related properties as revealed by geospatial analyses. Results show that P availability in the buffer was strongly related to soil genesis and support site-specific approaches for P loss risk evaluation in buffers. PMID:26828185

  14. DOM composition and lability during the Arctic spring freshet on the River Kolyma, Northeast Siberia

    NASA Astrophysics Data System (ADS)

    Mann, P. J.; Davydova, A. I.; Zimov, N.; Bulygina, E. B.; Davydov, S.; Russell-Roy, L.; Zimov, S. A.; Holmes, R. M.

    2010-12-01

    The concentration, composition, age and lability of dissolved organic matter (DOM) exported by Arctic Rivers is known to vary significantly across the hydrograph, affecting the biogeochemical cycling of carbon within high latitude systems. Despite the fact that the majority of the dissolved organic carbon (DOC) flux occurs during the spring freshet, surprisingly little is known about concurrent changes in DOM composition and how this may affect DOC removal processes. In this study, we collected a range of samples throughout the 2010 spring freshet in the Kolyma River and its nearby tributaries, Northeast Siberia. DOM quantity and quality was measured using absorbance and fluorescence spectrophotometry alongside bulk carbon (DOC) and nitrogen (TDN) measurements. Parallel factor analysis (PARAFAC) was employed to decompose fluorescence excitation-emission matrices (EEMs) into distinct fractions. In conjunction with DOM characterization, we investigated the photochemical and bacterial lability of the DOC pool using natural incubation experiments. Kolyma river DOC concentrations ranged from 0.6 mgL-1 under-ice to 14.2 mgL-1 at the peak of the spring flush. Up to 20% DOC loss was observed during 28 d-1 bacterial incubations and 13% loss during 4 d-1 natural sunlight incubations respectively. These percentage losses were greatest in association with maximum riverine DOC concentrations. Furthermore, prior sunlight exposure increased DOM availability for biological remineralization suggesting synergistic pathways in DOC degradation. Absorbance spectral slopes, SUVA254 and PARAFAC modeling results confirm significant changes in DOM composition during the flush and provide useful proxies for DOC lability. With predicted future changes in land-to-ocean carbon fluxes, understanding the link between DOM compositional changes and DOC photo and biolability has clear implications for the overall fate and export of carbon in high-latitude systems.

  15. Peat carbon stocks and potential microbial lability of boreal peatlands with varying permafrost histories

    NASA Astrophysics Data System (ADS)

    Olefeldt, D.; Pelletier, N.; Talbot, J.; Blodau, C.; Turetsky, M. R.

    2014-12-01

    Large stores of C in the form of peat are stored in permafrost, particularly in the boreal discontinuous permafrost zone. Ongoing climate change is causing widespread permafrost thaw in boreal peatlands, a trend which is expected to continue this century and thus make large stores of soil C available for microbial processes and mineralization. Permafrost thaw in boreal peatlands is often associated with an ecosystem shift from dry peat plateau to wet bog surfaces, and the net C balance following thaw is determined by the balance between the mineralization of plateau peat and the new accumulation of bog peat on top. In this study we collected soil cores (~3 m deep) from one peat plateaus and four bogs that differed in time since thaw (approximately 10, 50 and 500 years since thaw). In order to assess the potential microbial lability, we incubated 25 soil samples from each core under aerobic conditions at 17.5 deg C. Mineralization rates were 1-2 order of magnitude higher near the surface than at depth, but near surface samples also had high variability among cores. Variability in peat microbial lability near the surface was related to thaw history and to differences in characteristics between plateau and bog peat. Mineralization rates of peat samples from below 1 m depth and down to the interface with mineral soil at 3 m were consistently low and had no difference among cores. Mineralization rates during the first 3 months of incubation for deep plateau peat samples were equivalent to 1% soil C losses per year. Relatively low microbial lability of deep peat in combination with high rates of new peat accumulation during the initial stages of bog development suggests that there is net C accumulation immediately following thaw but that the sink strength weakens or reverses during later stages when new accumulation rates diminish.

  16. Conformational changes and loose packing promote E. coli Tryptophanase cold lability

    PubMed Central

    Kogan, Anna; Gdalevsky, Garik Y; Cohen-Luria, Rivka; Goldgur, Yehuda; Phillips, Robert S; Parola, Abraham H; Almog, Orna

    2009-01-01

    Background Oligomeric enzymes can undergo a reversible loss of activity at low temperatures. One such enzyme is tryptophanase (Trpase) from Escherichia coli. Trpase is a pyridoxal phosphate (PLP)-dependent tetrameric enzyme with a Mw of 210 kD. PLP is covalently bound through an enamine bond to Lys270 at the active site. The incubation of holo E. coli Trpases at 2C for 20 h results in breaking this enamine bond and PLP release, as well as a reversible loss of activity and dissociation into dimers. This sequence of events is termed cold lability and its understanding bears relevance to protein stability and shelf life. Results We studied the reversible cold lability of E. coli Trpase and its Y74F, C298S and W330F mutants. In contrast to the holo E. coli Trpase all apo forms of Trpase dissociated into dimers already at 25C and even further upon cooling to 2C. The crystal structures of the two mutants, Y74F and C298S in their apo form were determined at 1.9 resolution. These apo mutants were found in an open conformation compared to the closed conformation found for P. vulgaris in its holo form. This conformational change is further supported by a high pressure study. Conclusion We suggest that cold lability of E. coli Trpases is primarily affected by PLP release. The enhanced loss of activity of the three mutants is presumably due to the reduced size of the side chain of the amino acids. This prevents the tight assembly of the active tetramer, making it more susceptible to the cold driven changes in hydrophobic interactions which facilitate PLP release. The hydrophobic interactions along the non catalytic interface overshadow the effect of point mutations and may account for the differences in the dissociation of E. coli Trpase to dimers and P. vulgaris Trpase to monomers. PMID:19814824

  17. Interactions between recalcitrant and labile organic carbon in streams - Can stream biofilms mediate a priming effect?

    NASA Astrophysics Data System (ADS)

    Bengtsson, M. M.; Wagner, K.; Herberg, E. R.; Burns, N. R.; Wanek, W.; Battin, T. J.

    2012-04-01

    Inland waters - such as streams, rivers and lakes - are increasingly recognized as important components in the global carbon cycle. Dissolved organic carbon (DOC) in these systems is diverse in structure, origin and reactivity, and a fraction of it is regarded as recalcitrant to microbial degradation. In soils, degradation of recalcitrant carbon is often controlled by the availability of labile carbon sources. This is linked to the priming effect (PE). Mounting evidence suggests that PE is also important in aquatic ecosystems but there are so far very few studies addressing this topic. Biofilms are vital components of aquatic ecosystems. In stream biofilms, heterotrophic bacteria and algae coexist in close proximity, exposing the bacteria to both recalcitrant DOC of terrestrial origin and labile organic carbon from the algae. We hypothesize that this makes stream biofilms hotspots for PE. We used plug-flow bioreactors inoculated with natural stream biofilm bacterial communities to test the potential of a priming effect in aquatic ecosystems. The bioreactors were amended with an isotope-labeled plant extract serving as a model of recalcitrant DOC in streams. Labile carbon sources, in the form of glucose and an algal extract were added to induce PE. Nitrate and phosphate were also added to assess the role of these inorganic nutrients on carbon uptake. Microbial uptake of the different carbon sources was monitored by measuring the concentrations and isotopic ratios of respired CO2, biomass and DOC. Our results suggest that the priming effect plays a role in stream carbon cycling and that it is potentially an important process in other aquatic ecosystems.

  18. Dissolved organic carbon lability increases with water residence time in the alluvial aquifer of a river floodplain ecosystem

    NASA Astrophysics Data System (ADS)

    Helton, Ashley M.; Wright, Meredith S.; Bernhardt, Emily S.; Poole, Geoffrey C.; Cory, Rose M.; Stanford, Jack A.

    2015-04-01

    We assessed spatial and temporal patterns of dissolved organic carbon (DOC) lability and composition throughout the alluvial aquifer of the 16 km2 Nyack Floodplain in northwest Montana, USA. Water influx to the aquifer derives almost exclusively from the Middle Fork of the Flathead River, and water residence times within the aquifer range from days to months. Across seasons and channel discharge conditions, we measured DOC concentration, lability, and optical properties of aquifer water sampled from 12 wells, both near and ~3 m below the water table. Concentrations of DOC were typically low (542 ± 22.7 µg L-1; mean ± se), and the percentage of labile DOC averaged 18 ± 12% during 3 day laboratory assays. Parallel factor analysis of fluorescence excitation-emission matrices revealed two humic-like and two amino acid-like fluorescence groups. Total DOC, humic-like components, and specific UV absorbance decreased with water residence time, consistent with sorption to aquifer sediments. However, labile DOC (both concentration and fraction) increased with water residence time, suggesting a concurrent influx or production of labile DOC. Thus, although the carbon-poor, oxygen-rich aquifer is a net sink for DOC, recalcitrant DOC appears to be replaced with more labile DOC along aquifer flow paths. Our observation of DOC production in long flow paths contrasts with studies of hyporheic DOC consumption along short (centimeters to meters) flow paths and highlights the importance of understanding the role of labile organic matter production and/or influx in alluvial aquifer carbon cycling.

  19. Enantiomeric Excesses of Acid Labile Amino Acid Precursors of the Murchison Meteorite

    NASA Technical Reports Server (NTRS)

    Pizzarello, Sandra

    1998-01-01

    Amino acids present in carbonaceous chondrite are extracted in water in part as free compounds and in approximately equal part as acid labile precursors. On the assumption that they would be free of contamination, the precursors of two Murchison amino acids that have terrestrial occurrence, alanine and glutamic acid, have been targeted for analysis of their enantiomeric ratios. Pyroglutamic acid, the precursor of glutamic acid, was found with an L-enantiomeric excess comparable to that of the free acid, while alanine's precursor, N-acetyl alanine, appears approximately racemic. Also alpha-imino propioacetic acid, a proposed end product of alanine synthesis in the meteorite, was analyzed and found racemic.

  20. Epithelial to mesenchymal transition-the roles of cell morphology, labile adhesion and junctional coupling.

    PubMed

    Abdulla, Tariq; Luna-Zurita, Luis; de la Pompa, Jos Luis; Schleich, Jean-Marc; Summers, Ron

    2013-08-01

    Epithelial to mesenchymal transition (EMT) is a fundamental process during development and disease, including development of the heart valves and tumour metastases. An extended cellular Potts model was implemented to represent the behaviour emerging from autonomous cell morphology, labile adhesion, junctional coupling and cell motility. Computer simulations normally focus on these functional changes independently whereas this model facilitates exploration of the interplay between cell shape changes, adhesion and migration. The simulation model is fitted to an in vitro model of endocardial EMT, and agrees with the finding that Notch signalling increases cell-matrix adhesion in addition to modulating cell-cell adhesion. PMID:23787029

  1. Reductively labile PRINT particles for the delivery of doxorubicin to HeLa cells.

    PubMed

    Petros, Robby A; Ropp, Patricia A; DeSimone, Joseph M

    2008-04-16

    A Trojan horse PRINT particle composition was developed that incorporates a reductively labile cross-linker to achieve activated release of doxorubicin in vitro. Particles of discrete size and shape (cube side length = 2 micron) containing 30 wt % of a disulfide-based cross-linker and 2 wt % doxorubicin were synthesized. This PRINT composition was shown to release doxorubicin in response to a reducing environment as measured by flow cytometry and was found to be highly proficient at killing HeLa cells in vitro. PMID:18355010

  2. Investigating the role of solanapyrone toxins in Ascochyta blight using toxin-deficient mutants of Asochyta rabiei

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ascochyta rabiei, the causal agent of Ascochyta blight of chickpea, produces solanapyrone toxins (solanapyrone A, B and C). However, very little is known about the genetics of toxin production and the role of the toxins in pathogenesis. Generating mutants deficient in the toxin biosynthesis would p...

  3. Inhibition of Shiga toxin 2 (Stx2) in apple juices and its resistance to pasteurization.

    PubMed

    Rasooly, Reuven; Do, Paula M; Levin, Carol E; Friedman, Mendel

    2010-06-01

    In the present study, we evaluated Shiga toxin (Stx2) activity in apple juices by measuring a decrease in dehydrogenase activity of Vero cells with the microculture tetrazolium (MTT) assay. Freshly prepared juice from Red Delicious apples and Golden Delicious apples inhibited the biological activity of the bacterial toxin Stx2 produced by E. coli O157:H7 strains. Studies with immunomagnetic beads bearing specific antibodies against the toxin revealed that Stx2 activity was restored when removed from the apple juice. SDS gel electrophoresis revealed no difference (P < 0.05) in the densities or molecular weights between Stx2 in either PBS or apple juices. These results suggest that Stx2 may be reversibly bound to small molecular weight constituents in the juice. The Stx2 toxin was not inactivated on exposure to heat programs (63 degrees C for 30 min, 72 degrees C for 15 s, 89 degrees C for 1 s) commonly used to pasteurize apple juice, but lost all activity when exposed to 100 degrees C for 5 min. The results suggest that pasteurization of apple juice used to inactivate E. coli O157:H7 has no effect on Stx2, and that food-compatible and safe antitoxin compounds can be used to inhibit the biological activity of the Shiga toxin. PMID:20629887

  4. New players in the toxin field: polymorphic toxin systems in bacteria.

    PubMed

    Jamet, Anne; Nassif, Xavier

    2015-01-01

    Bacteria have evolved numerous strategies to increase their competitiveness and fight against each other. Indeed, a large arsenal of antibacterial weapons is available in order to inhibit the proliferation of competitor cells. Polymorphic toxin systems (PTS), recently identified by bioinformatics in all major bacterial lineages, correspond to such a system primarily involved in conflict between related bacterial strains. They are typically composed of a secreted multidomain toxin, a protective immunity protein, and multiple cassettes encoding alternative toxic domains. The C-terminal domains of polymorphic toxins carry the toxic activity, whereas the N-terminal domains are related to the trafficking mode. In silico analysis of PTS identified over 150 distinct toxin domains, including putative nuclease, deaminase, or peptidase domains. Immunity genes found immediately downstream of the toxin genes encode small proteins that protect bacteria against their own toxins or against toxins secreted by neighboring cells. PTS encompass well-known colicins and pyocins, contact-dependent growth inhibition systems which include CdiA and Rhs toxins and some effectors of type VI secretion systems. We have recently characterized the MafB toxins, a new family of PTS deployed by pathogenic Neisseria spp. Many other putative PTS have been identified by in silico predictions but have yet to be characterized experimentally. However, the high number of these systems suggests that PTS have a fundamental role in bacterial biology that is likely to extend beyond interbacterial competition. PMID:25944858

  5. New Players in the Toxin Field: Polymorphic Toxin Systems in Bacteria

    PubMed Central

    Nassif, Xavier

    2015-01-01

    ABSTRACT Bacteria have evolved numerous strategies to increase their competitiveness and fight against each other. Indeed, a large arsenal of antibacterial weapons is available in order to inhibit the proliferation of competitor cells. Polymorphic toxin systems (PTS), recently identified by bioinformatics in all major bacterial lineages, correspond to such a system primarily involved in conflict between related bacterial strains. They are typically composed of a secreted multidomain toxin, a protective immunity protein, and multiple cassettes encoding alternative toxic domains. The C-terminal domains of polymorphic toxins carry the toxic activity, whereas the N-terminal domains are related to the trafficking mode. In silico analysis of PTS identified over 150 distinct toxin domains, including putative nuclease, deaminase, or peptidase domains. Immunity genes found immediately downstream of the toxin genes encode small proteins that protect bacteria against their own toxins or against toxins secreted by neighboring cells. PTS encompass well-known colicins and pyocins, contact-dependent growth inhibition systems which include CdiA and Rhs toxins and some effectors of type VI secretion systems. We have recently characterized the MafB toxins, a new family of PTS deployed by pathogenic Neisseria spp. Many other putative PTS have been identified by in silico predictions but have yet to be characterized experimentally. However, the high number of these systems suggests that PTS have a fundamental role in bacterial biology that is likely to extend beyond interbacterial competition. PMID:25944858

  6. Bacterial Toxins as Pathogen Weapons Against Phagocytes.

    PubMed

    do Vale, Ana; Cabanes, Didier; Sousa, Sandra

    2016-01-01

    Bacterial toxins are virulence factors that manipulate host cell functions and take over the control of vital processes of living organisms to favor microbial infection. Some toxins directly target innate immune cells, thereby annihilating a major branch of the host immune response. In this review we will focus on bacterial toxins that act from the extracellular milieu and hinder the function of macrophages and neutrophils. In particular, we will concentrate on toxins from Gram-positive and Gram-negative bacteria that manipulate cell signaling or induce cell death by either imposing direct damage to the host cells cytoplasmic membrane or enzymatically modifying key eukaryotic targets. Outcomes regarding pathogen dissemination, host damage and disease progression will be discussed. PMID:26870008

  7. New indications for botulinum toxin in rheumatology.

    PubMed

    Monnier, Guy; Tatu, Laurent; Michel, Fabrice

    2006-12-01

    Previously known only as a deadly bacterial poison responsible for severe paralysis, botulinum toxin is now a well-recognized therapeutic agent used to relieve involuntary movements, dystonia-related functional impairments, spasticity, and autonomic disorders such as hyperhidrosis. Musculoskeletal pain in patients with rheumatic disorders is among the emerging indications for botulinum toxin therapy. Preliminary data have been obtained in patients with cervical or thoracolumbar myofascial pain syndrome, chronic low back pain, piriformis muscle syndrome, tennis elbow, and stiff person syndrome. At present, the effects of botulinum toxin and its use for pain relief remain controversial. Carefully designed prospective trials are needed to investigate the efficacy and safety of botulinum toxin in pain disorders. PMID:16997603

  8. How Parkinsonian Toxins Dysregulate the Autophagy Machinery

    PubMed Central

    Dagda, Ruben K.; Das Banerjee, Tania; Janda, Elzbieta

    2013-01-01

    Since their discovery, Parkinsonian toxins (6-hydroxydopamine, MPP+, paraquat, and rotenone) have been widely employed as in vivo and in vitro chemical models of Parkinson’s disease (PD). Alterations in mitochondrial homeostasis, protein quality control pathways, and more recently, autophagy/mitophagy have been implicated in neurotoxin models of PD. Here, we highlight the molecular mechanisms by which different PD toxins dysregulate autophagy/mitophagy and how alterations of these pathways play beneficial or detrimental roles in dopamine neurons. The convergent and divergent effects of PD toxins on mitochondrial function and autophagy/mitophagy are also discussed in this review. Furthermore, we propose new diagnostic tools and discuss how pharmacological modulators of autophagy/mitophagy can be developed as disease-modifying treatments for PD. Finally, we discuss the critical need to identify endogenous and synthetic forms of PD toxins and develop efficient health preventive programs to mitigate the risk of developing PD. PMID:24217228

  9. INVESTIGATOIN OF CYANOBACTERIA TOXINS IN WATER

    EPA Science Inventory

    Introduction:

    Approximately 80 alkaloid and cyclic peptide toxins produced by various freshwater and marine cyanobacteria (blue-green algae) have been identified and their structures determined. The U. S. Environmental Protection Agency has identified two neurotoxin alkalo...

  10. Biosynthetic studies of the DSP toxin skeleton.

    PubMed

    Daranas, Antonio H; Fernndez, Jos J; Norte, Manuel; Gavn, Jos A; Surez-Gmez, Blanca; Souto, Mara L

    2004-01-01

    Marine toxins have drawn wide interest because their economical impact and disastrous effect upon the shellfish industry and public health in many parts of the world. One of the most interesting group of substances of marine toxins, from structural and pharmacological points of view are polyether compounds, which generally present a great diversity in size and potent biological activities. The subject of this work was about to biosynthesis of okadaic acid skeleton as leader as DSP toxins. Its biosynthesis attracts considerable attention since the carbon skeleton has been shown to be synthesised via an unusual route. In this paper we report on stable isotope incorporation experiments on DSP toxin in artificial cultures of dinoflagellate. The comparison of the degrees of incorporation in these samples measured by different methods led to contradictory results. This implies that further experimental data is needed in order to propose a logical biogenetic scheme. PMID:15057864

  11. Botulinum toxin type a for chronic migraine.

    PubMed

    Ashkenazi, Avi

    2010-03-01

    Chronic migraine (CM) is the leading cause of chronic daily headache, a common and debilitating headache syndrome. The management of CM patients is challenging, with only limited benefit from available oral preventive medications. Botulinum neurotoxin (BoNT) has been used extensively to treat disorders associated with increased muscle tone. More recent scientific data support an analgesic effect of the toxin. The pharmacokinetic and pharmacodynamic profiles of BoNT make it an appealing candidate for migraine prevention. Results from older clinical trials on the efficacy of the toxin in CM were inconclusive. However, recent trials using more stringent inclusion criteria have shown positive results, supporting the use of the toxin in some patients with this disorder. This review summarizes the scientific data on the analgesic properties of BoNT, as well as the clinical data on the efficacy of the toxin in treating CM. PMID:20425239

  12. Bacterial Toxins as Pathogen Weapons Against Phagocytes

    PubMed Central

    do Vale, Ana; Cabanes, Didier; Sousa, Sandra

    2016-01-01

    Bacterial toxins are virulence factors that manipulate host cell functions and take over the control of vital processes of living organisms to favor microbial infection. Some toxins directly target innate immune cells, thereby annihilating a major branch of the host immune response. In this review we will focus on bacterial toxins that act from the extracellular milieu and hinder the function of macrophages and neutrophils. In particular, we will concentrate on toxins from Gram-positive and Gram-negative bacteria that manipulate cell signaling or induce cell death by either imposing direct damage to the host cells cytoplasmic membrane or enzymatically modifying key eukaryotic targets. Outcomes regarding pathogen dissemination, host damage and disease progression will be discussed. PMID:26870008

  13. Nanoanalysis of the arthropod neuro-toxins

    PubMed Central

    Nakajima, Terumi

    2006-01-01

    Many kinds of venomous principles modulate physiological responses of mammalian signal transduction systems, on which they act selectively as enhancers, inhibitors or some other kind of effectors. These toxins become useful tools for physiological research. We have employed and characterized paralyzing toxins from the venom of spiders, insects and scorpions with a limited supply. We have developed rapid and sensitive mass spectrometric technology and applied for the identification of these toxins. Venom profiles are screened by MALDI-TOF fingerprinting analysis prior to purification of venomous components, then marked target toxins of small molecular mass (10005000) are characterized directly by means of mass spectrometric techniques such as Frit-FAB MS/MS, CID/PSD-TOF MS, Capil.-HPLC/Q-TOF MS/MS etc. PMID:25792792

  14. Relationship between the lability of sediment-bound metals (Cd, Cu, Zn) and their bioaccumulation in benthic invertebrates

    NASA Astrophysics Data System (ADS)

    Amiard, J.-C.; Geffard, A.; Amiard-Triquet, C.; Crouzet, C.

    2007-04-01

    The present study has investigated metal contamination at nine sites (10 sampling stations) from the English Channel to the Mediterranean Sea, including low level and highly contaminated sediments. Both total and labile concentrations of metals were determined in superficial sediments. The influence of different pHs was tested and metal lability at pHs encountered in the gut of invertebrates (the ragworm Nereis diversicolor, the blue mussel Mytilus edulis, the Japanese oyster Crassostrea gigas) was compared with the distribution of metals in various operationally defined geochemical fractions. Cd showed the highest lability and Cu the lowest, whereas Zn lability was intermediate. Metal concentrations were determined in bivalves at six sites and in worms at three sites. Cd in living organisms and labile Cd in sediments increased in proportion over the gradient of contamination. This relationship did not always hold for Cu and Zn and these exceptions are discussed. Even if sediments are not the only source of metal contamination in marine invertebrates, the procedure proposed here to assess metal bioavailability by remobilising sediment-bound metals at physiological pHs, seems a significant improvement of the existing methodologies of risk assessment.

  15. Dermatitis from purified sea algae toxin (debromoaplysiatoxin).

    PubMed

    Solomon, A E; Stoughton, R B

    1978-09-01

    Cutaneous inflammation was induced by debromoaplysiatoxin, a purified toxin extracted from Lyngbya majuscula Gomont. This alga causes a seaweed dermatitis that occurs in persons who have swum off the coast of Oahu in Hawaii. By topical application, the toxin was found to produce an irritant pustular folliculitis in humans and to cause a severe cutaneous inflammatory reaction in the rabbit and in hairless mice. PMID:686747

  16. [Axillary hyperhidrosis, botulinium A toxin treatment: Review].

    PubMed

    Clerico, C; Fernandez, J; Camuzard, O; Chignon-Sicard, B; Ihrai, T

    2016-02-01

    Injection of type A botulinum toxin in the armpits is a temporary treatment for axillary hyperhidrosis. This technique described in 1996 by Bushara et al., is known to be efficient and safe. The purpose of this article was to review the data concerning the treatment of axillary hyperhidrosis with botulinum toxin type A, and discuss the other treatment modalities for this socially disabling entity. PMID:25555435

  17. A labile hydride strategy for the synthesis of heavily nitridized BaTiO3.

    PubMed

    Yajima, Takeshi; Takeiri, Fumitaka; Aidzu, Kohei; Akamatsu, Hirofumi; Fujita, Koji; Yoshimune, Wataru; Ohkura, Masatoshi; Lei, Shiming; Gopalan, Venkatraman; Tanaka, Katsuhisa; Brown, Craig M; Green, Mark A; Yamamoto, Takafumi; Kobayashi, Yoji; Kageyama, Hiroshi

    2015-12-01

    Oxynitrides have been explored extensively in the past decade because of their interesting properties, such as visible-light absorption, photocatalytic activity and high dielectric permittivity. Their synthesis typically requires high-temperature NH3 treatment (800-1,300 °C) of precursors, such as oxides, but the highly reducing conditions and the low mobility of N(3-) species in the lattice place significant constraints on the composition and structure-and hence the properties-of the resulting oxynitrides. Here we show a topochemical route that enables the preparation of an oxynitride at low temperatures (<500 °C), using a perovskite oxyhydride as a host. The lability of H(-) in BaTiO3-xHx (x ≤ 0.6) allows H(-)/N(3-) exchange to occur, and yields a room-temperature ferroelectric BaTiO3-xN2x/3. This anion exchange is accompanied by a metal-to-insulator crossover via mixed O-H-N intermediates. These findings suggest that this 'labile hydride' strategy can be used to explore various oxynitrides, and perhaps other mixed anionic compounds. PMID:26587718

  18. Constraints on Transport and Emplacement Mechanisms of Labile Fractions in Lunar Cold Traps

    NASA Technical Reports Server (NTRS)

    Rickman, D.; Gertsch, L.

    2014-01-01

    Sustaining the scientific exploration of the Solar System will require a significant proportion of the necessary fuels and propellants, as well as other bulk commodities, to be produced from local raw materials [1]. The viability of mineral production depends on the ability to locate and characterize mineable deposits of the necessary feedstocks. This requires, among other things, a workable understanding of the mechanisms by which such deposits form, which is the subject of Economic Geology. Multiple deposition scenarios are possible for labile materials on the Moon. This paper suggests labile fractions moved diffusely through space; deposits may grow richer with depth until low porosity rock; lateral transport is likely to have occurred with the regolith, at least for short distances; crystalline ice may not exist; the constituent phases could be extremely complex. At present we can constrain the sources only mildly; once on the Moon, the transport mechanisms inherently mix and therefore obscure the origins. However, the importance of expanding our understanding of ore-forming processes on the Moon behooves us to make the attempt. Thus begins a time of new inquiry for Economic Geology.

  19. A labile hydride strategy for the synthesis of heavily nitridized BaTiO3

    NASA Astrophysics Data System (ADS)

    Yajima, Takeshi; Takeiri, Fumitaka; Aidzu, Kohei; Akamatsu, Hirofumi; Fujita, Koji; Yoshimune, Wataru; Ohkura, Masatoshi; Lei, Shiming; Gopalan, Venkatraman; Tanaka, Katsuhisa; Brown, Craig M.; Green, Mark A.; Yamamoto, Takafumi; Kobayashi, Yoji; Kageyama, Hiroshi

    2015-12-01

    Oxynitrides have been explored extensively in the past decade because of their interesting properties, such as visible-light absorption, photocatalytic activity and high dielectric permittivity. Their synthesis typically requires high-temperature NH3 treatment (8001,300?C) of precursors, such as oxides, but the highly reducing conditions and the low mobility of N3? species in the lattice place significant constraints on the composition and structureand hence the propertiesof the resulting oxynitrides. Here we show a topochemical route that enables the preparation of an oxynitride at low temperatures (<500?C), using a perovskite oxyhydride as a host. The lability of H? in BaTiO3?xHx (x ? 0.6) allows H?/N3? exchange to occur, and yields a room-temperature ferroelectric BaTiO3?xN2x/3. This anion exchange is accompanied by a metal-to-insulator crossover via mixed OHN intermediates. These findings suggest that this labile hydride strategy can be used to explore various oxynitrides, and perhaps other mixed anionic compounds.

  20. Localization of alkali-labile sites in donkey (Equus asinus) and stallion (Equus caballus) spermatozoa.

    PubMed

    Corts-Gutirrez, Elva I; Dvila-Rodrguez, Martha I; Lpez-Fernndez, Carmen; Fernndez, Jos Luis; Crespo, Francisco; Goslvez, Jaime

    2014-01-15

    The presence of constitutive alkali-labile sites (ALS) has been investigated using a protocol of DNA breakage detection-fluorescence in situ hybridization and comet assay in spermatozoa of donkey (Equus asinus) and stallion (Equus caballus). These results were compared with those obtained using a similar experimental approach using somatic cells. The relative abundance of ALS was of the order of four times more in spermatozoa than in somatic cells. Alkali-labile sites showed a tendency to cluster localized at the equatorial-distal regions of the sperm. The amount of hybridized signal in the ALS in the sperm of donkey (Equus asinus) was 1.3 times greater than in stallion (Equus caballus), and the length of the comet tail obtained in donkey sperm was 1.6 times longer than that observed in stallion (P < 0.05); however, these differences were not appreciated in somatic cells. In conclusion, ALS localization in sperm is not a randomized event and a different pattern of ALS distribution occurs for each species. These results suggest that ALS represents a species-specific issue related to chromatin organization in sperm and somatic cells in mammalian species, and they might diverge even with very short phylogenetic distances. PMID:24182740

  1. High rates of soil respiration suggest large fluxes of labile C in a turfgrass system

    NASA Astrophysics Data System (ADS)

    Lilly, P.; Jenkins, J. C.; Carroll, M.

    2013-12-01

    We measured high rates of soil respiration (up to 19 ?mol C m-2 sec-1) in a study of management effects on turfgrass lawns. Estimates based on these measurements indicate that annual soil respiration is on the order of 15-30 Mg C ha-1, which seems high, given measured NPP of 3-6 Mg C ha-1 yr-1 and 0-10 cm soil C pools of 15-20 Mg C ha-1 which showed no net change over the study period. We contend that this flux is plausible, given irrigation and fertilizer applications, and hypothesize that in this context, soil respiration is driven largely by assimilate availability. We provide several lines of evidence supporting the idea that large fluxes of readily labile C move through this system: Biomass growth rates were highly correlated with soil respiration rates; we observed statistically significant variation in soil C stocks, dramatic changes in microbial biomass stoichiometry, and major shifts in N mineralization and nitrification over the course of a single growing season; and long-term soil incubations appear to show the accumulation and depletion of a substantial pool of labile C.

  2. Lack of correlation between non-labile iron parameters, total carbonyl and malondialdehyde in major thalassemia

    PubMed Central

    Al-Hakeim, Hussein Kadhem; Auda, Furqan Muein; Ali, Basim Muhammed

    2014-01-01

    Thalassemia patients are at high risk of iron-induced toxicity and oxidative stress consequences. The present cross-sectional study is conducted to determine whether or not lipid peroxidation or protein oxidation is correlated with iron parameters in patients with thalassemia major. To prove this hypothesis, malondialdehyde and total carbonyl were correlated with the degree of excess iron concentration in the patients. A total of 118 Arabic Iraqi patients and 30 healthy children were participated in the present study. Results showed a significant increase (p<0.05) in serum total carbonyls, malondialdehyde and the iron indices of patients as compared with the control group. Total iron binding capacity and transferrin concentrations decreased significantly (p<0.05) in patients with thalassemia compared with the control group. The results also showed a lack of a significant correlation between each serum malondialdehyde and total carbonyl with each component of iron status. In conclusion, total carbonyls and malondialdehyde were increased in thalassemia patients indicating the vulnerability of these patients to tissue injury caused by oxidative stress. The formation of total carbonyl and malondialdehyde are independent of excess non-labile iron concentration, indicating that different mechanisms are involved in injury caused by the labile iron and in the formation of oxidation end products. PMID:25411527

  3. Rhizosphere Environment and Labile Phosphorus Release from Organic Waste-Amended Soils.

    NASA Astrophysics Data System (ADS)

    Dao, Thanh H.

    2015-04-01

    Crop residues and biofertilizers are primary sources of nutrients for organic crop production. However, soils treated with large amounts of nutrient-enriched manure have elevated phosphorus (P) levels in regions of intensive animal agriculture. Surpluses occurred in these amended soils, resulting in large pools of exchangeable inorganic P (Pi) and enzyme-labile organic P (Po) that averaging 30.9 and 68.2 mg kg-1, respectively. Organic acids produced during crop residue decomposition can promote the complexation of counter-ions and decouple and release unbound Pi from metal and alkali metal phosphates. Animal manure and cover crop residues also contain large amounts of soluble organic matter, and likely generate similar ligands. However, a high degree of heterogeneity in P spatial distribution in such amended fields, arising from variances in substrate physical forms ranging from slurries to dried solids, composition, and diverse application methods and equipment. Distinct clusters of Pi and Po were observed, where accumulation of the latter forms was associated with high soil microbial biomass C and reduced phosphomonoesterases' activity. Accurate estimates of plant requirements and lability of soil P pools, and real-time plant and soil P sensing systems are critical considerations to optimally manage manure-derived nutrients in crop production systems. An in situ X-ray fluorescence-based approach to sensing canopy and soil XRFS-P was developed to improve the yield-soil P relationship for optimal nutrient recommendations in addition to allowing in-the-field verification of foliar P status.

  4. Interaction of macrophage migration inhibitory factor with ceruloplasmin: role of labile copper ions.

    PubMed

    Kostevich, Valeria A; Sokolov, Alexey V; Grudinina, Natalia A; Zakharova, Elena T; Samygina, Valeria R; Vasilyev, Vadim B

    2015-10-01

    Macrophage migration inhibitory factor (MIF), a pro-inflammatory cytokine, is a target for pharmacological treatment of sepsis and malignant tumors. Inhibition of tautomerase activity of MIF in reaction with p-hydroxyphenylpyruvate (HPP) was observed in the presence of ceruloplasmin (CP), a copper-containing plasma protein. Binding labile copper ions to CP (CP+Cu(II)) is a prerequisite for MIF inhibiting. CP+Cu(II) is shown to be an uncompetitive inhibitor of MIF (Ki ~ 37 nM), which suggests formation of a complex 'MIF-HPP-CP-Cu(II)'. Filtration of CP+Cu(II) on a column with Chelex-100, otherwise the presence of high concentrations of histidine, cysteine or methionine abrogated the inhibitory effect of CP. Adding salts of Co(II) and Ni(II) that replace copper ions in the labile sites prevented the inhibitory effect of CP+Cu(II). Limited proteolysis of CP by thrombin diminished its oxidase activity in reaction with p-phenylenediamine, but endowed it with the capacity of inhibiting MIF. Covalent modification of MIF by phenylmethylsulfonyl fluoride (PMSF) resulted in binding of MIF-PMSF to CP immobilized on CM5 chip, the dissociation constant being 4.2 μM. In D-galactosamine-sensitized mice CP+Cu(II) increased the LPS-induced lethality from 54 to 100%, while administration of antibodies against MIF prevented the lethal effect. The enhancement by CP+Cu(II) of the pro-inflammatory signal of MIF is discussed. PMID:26091949

  5. Tetra- versus Pentavalent Inhibitors of Cholera Toxin**

    PubMed Central

    Fu, Ou; Pukin, Aliaksei V; van?Ufford, H C Quarles; Branson, Thomas R; Thies-Weesie, Dominique M E; Turnbull, W Bruce; Visser, Gerben M; Pieters, Roland J

    2015-01-01

    The five B-subunits (CTB5) of the Vibrio cholerae (cholera) toxin can bind to the intestinal cell surface so the entire AB5 toxin can enter the cell. Simultaneous binding can occur on more than one of the monosialotetrahexosylganglioside (GM1) units present on the cell surface. Such simultaneous binding arising from the toxins multivalency is believed to enhance its affinity. Thus, blocking the initial attachment of the toxin to the cell surface using inhibitors with GM1 subunits has the potential to stop the disease. Previously we showed that tetravalent GM1 molecules were sub-nanomolar inhibitors of CTB5. In this study, we synthesized a pentavalent version and compared the binding and potency of penta- and tetravalent cholera toxin inhibitors, based on the same scaffold, for the first time. The pentavalent geometry did not yield major benefits over the tetravalent species, but it was still a strong inhibitor, and no major steric clashes occurred when binding the toxin. Thus, systems which can adopt more geometries, such as those described here, can be equally potent, and this may possibly be due to their ability to form higher-order structures or simply due to more statistical options for binding. PMID:26478842

  6. Analysis of remineralisation, lability, temperature sensitivity and structural composition of organic matter from the upper ocean

    NASA Astrophysics Data System (ADS)

    Bendtsen, Jrgen; Hilligse, Karen Marie; Hansen, Jrgen L. S.; Richardson, Katherine

    2015-01-01

    Organic carbon (OC) synthesised by plankton is exported out of the surface layer as particulate (POC) and dissolved (DOC) organic carbon. This 'biological pump' constitutes a major pathway in the global marine carbon cycle, each year exporting about 10 Pg C into the ocean interior, where it is subsequently remineralised via biological decomposition. Remineralised inorganic nutrients and carbon are, ultimately, again brought to the surface by advection and turbulent mixing which closes the OC-cycle in the upper ocean. Thus, remineralisation rates of OC are a critical component of the biological pump. These rates are regulated by the lability of the material and the environmental conditions in the ambient water. Temperature is particularly important in regulating the rate of microbial respiration and, thus, remineralisation rates. A significant temperature dependence of the microbial metabolic activity in the ocean interior is expected, as this is a feature observed elsewhere in the biosphere. Such temperature dependence of microbial remineralisation of POC and DOC will alter the amount of material available for transport by the biological pump to the deep ocean. Very few studies on the lability of OC and temperature sensitivity of microbial degradation processes in the mesopelagic zone (?100-1000 m) have, to date, been carried out. Here, we present a comprehensive new experimental data set from all major ocean basins and quantify remineralisation rates of OC and their temperature sensitivity in long-term incubations of water from the upper 350 m. Microbial respiration was measured by non-invasive oxygen optodes and oxygen consumption was used as a constraint for determining the remineralisation rates and temperature sensitivity by two complementary methods. First, we analysed the oxygen consumption from a multi-component OC-model where the concentration, remineralisation rates and temperature sensitivity of two bio-available (labile) pools of organic carbon were fitted to the data via a non-linear fitting procedure. Thereafter, a continuous OC-model was fitted to the data through an inverse method and information about lability, temperature sensitivity and structural composition of the OC-pool was analysed together with the results from the two-pool solutions. Median values of remineralisation rates from all experiments on material characterising sinking POC were found to be 0.6 and 0.05 days-1 for the two decomposable pools corresponding to turnover times of 2 and 21 days, respectively. Accordingly, solutions from the continuous model resulted in median turnover times between 6 and 11 days. Similar analyses were carried out for the OC-pool characterising DOC. A significant bio-available OC-pool was found to be present in the surface layer with a median value from all experiments of 30 ?M TOC. The median values of all remineralisation rates from the two bio-available OC-pools were found to be 0.2 and 0.02 days-1, corresponding to turnover times of 5 and 56 days, respectively, in good agreement with previous studies. The corresponding temperature sensitivities, characterised by a Q10-value, were found to be about 1.9 for the POC-fraction whereas the DOC fraction was characterised with values above 2.8 for the continuous OC-models. The analysis of the structural composition indicated that the TOC distribution in the surface layer was characterised by more heterogeneous material in terms of lability compared with the POC material sampled from the upper 350 m. Finally, we analyse the potential impact of the calculated temperature sensitivity on the general OC-cycling in the upper ocean and show that the implied reduction in OC-flux in a warmer ocean may have significant impact on nutrient cycling in general and also tends to reduce future ocean carbon uptake significantly.

  7. Metal contents of phytoplankton and labile particulate material in the North Atlantic Ocean

    NASA Astrophysics Data System (ADS)

    Twining, Benjamin S.; Rauschenberg, Sara; Morton, Peter L.; Vogt, Stefan

    2015-09-01

    Phytoplankton contribute significantly to global C cycling and serve as the base of ocean food webs. Phytoplankton require trace metals for growth and also mediate the vertical distributions of many metals in the ocean. We collected bulk particulate material and individual phytoplankton cells from the upper water column (<150 m) of the North Atlantic Ocean as part of the US GEOTRACES North Atlantic Zonal Transect cruise (GEOTRACES GA03). Particulate material was first leached to extract biogenic and potentially-bioavailable elements, and the remaining refractory material was digested in strong acids. The cruise track spanned several ocean biomes and geochemical regions. Particulate concentrations of metals associated primarily with lithogenic phases (Fe, Al, Ti) were elevated in surface waters nearest North America, Africa and Europe, and elements associated primarily with biogenic material (P, Cd, Zn, Ni) were also found at higher concentrations near the coasts. However metal/P ratios of labile particulate material were also elevated in the middle of the transect for Fe, Ni, Co, Cu, and V. P-normalized cellular metal quotas measured with synchrotron X-ray fluorescence (SXRF) were generally comparable to ratios in bulk labile particles but did not show mid-basin increases. Manganese and Fe ratios and cell quotas were higher in the western part of the section, nearest North America, and both elements were more enriched in bulk particles, relative to P, than in cells, suggesting the presence of labile oxyhydroxide particulate phases. Cellular Fe quotas thus did not increase in step with aeolian dust inputs, which are highest near Africa; these data suggest that the dust inputs have low bioavailability. Copper and Ni cell quotas were notably higher nearest the continental margins. Overall mean cellular metal quotas were similar to those measured in the Pacific and Southern Oceans except for Fe, which was approximately 3-fold higher in North Atlantic cells. Cellular Fe quotas are in-line with those measured in laboratory cultures at comparable Fe concentrations. Particulate Zn, Cu, Ni, and Co are primarily associated with cellular material, but less than 30% of labile particulate Fe and Mn are biogenic. Particulate Al was primarily associated with lithogenic material, but the labile fraction was highly correlated with P, as well as with biogenic silica, suggesting that some particulate Al (perhaps around 20%) may occur adsorbed to biogenic material. Cellular element maps indicate that externally scavenged Fe was not a significant fraction of the metal associated with live phytoplankton, but adsorbed or precipitated phases are likely to be important in particulate detrital material. Such abiotic scavenging, along with differential remineralization of cellular nutrients in the water column, results in estimates of cellular metal/nutrient ratios from dissolved concentrations that significantly underestimate the ratios in phytoplankton. These data demonstrate the response of phytoplankton to the unique metal inputs to the North Atlantic Ocean.

  8. Cu lability and bioavailability in an urban stream during baseflow versus stormflow

    NASA Astrophysics Data System (ADS)

    Vadas, T.; Luan, H.

    2012-12-01

    Urban streams are dynamic systems with many anthropogenic inputs and stressors. Existing contaminant inputs are regulated through total maximum daily loads. Techniques for assessing that load are based on a combination of acute and chronic water quality criteria, biotic ligand models, and physical, chemical and biological assessments. In addition, the apportionment of reduction in load to different sources is based on total mass and not, for example, on bioavailable fraction. Our understanding of the impact of different metal inputs to stream impairment is limited. Free metal ions are understood to play a role in direct cellular uptake, but metal speciation (e.g. free metal, labile metals, or size fractionated) is relevant to more complex stream food webs. As part of an ongoing study, this work examines dissolved and particulate Cu concentrations in the Hockanum River, Vernon, CT situated in a developed watershed. Stream samples were taken during baseflow as well as stormflow upstream and downstream of wastewater treatment plant and stormwater runoff inputs. In addition, diffusive gradient in thin-film (DGT) devices which measure labile metal concentrations and cultured periphyton were used to examine bioavailable fractions. Total and filtered Cu concentrations ranged from about 1.3 to 10.7 μg/L, and 0.9 to 5.1 μg/L, respectively. Cu concentrations always increased downstream of the wastewater treatment plant by about 1.1-2 times, and effluent accounted for about 30% of baseflow. Generally, small increases (<10%) in concentration were observed in metals directly downstream of stormwater inlets, likely due to low volumes of runoff contributed from stormwater outfalls during these sampling periods. However, Cu concentrations were elevated (about 2-5 times higher) at all sites downstream from the wastewater treatment plant downstream sampling point, suggesting contributions from sediment resuspension. DGT measured concentrations represented 30 to 70% of dissolved Cu concentrations, and that percentage increased in the days following a storm, suggesting more labile Cu compounds remained in the water column longer. Whereas solution metal concentrations in stormwater influenced reaches did not largely change upstream versus downstream, concentrations of Cu in periphyton cultures increased 2-fold downstream of a stormwater input. This is likely due to differences in discrete sampling locations, but indicates the potential importance of resuspended sediment on enhancing metal bioavailability. These results suggest that legacy sediment contamination is contributing to water column Cu concentrations. Downstream Cu concentrations are elevated compared to near the wastewater treatment plant effluent but are mostly unchanged before and after stormwater inputs. In addition, DGT concentrations indicate a shift in percentage of labile metals, in part distinguished based on size exclusion by the DGT device, following a storm event. As total dissolved Cu concentration decreases, labile Cu increases likely due to settling or size dependent partitioning to the solid phases such as periphyton. Understanding the transport and bioavailability of size dependent Cu compounds in stream water is important to allocating resources to mitigate metal contamination.

  9. ADP-ribosylation by cholera toxin: functional analysis of a cellular system that stimulates the enzymic activity of cholera toxin fragment A/sub 1/

    SciTech Connect

    Gill, D.M.; Coburn, J.

    1987-10-06

    The authors have clarified relationships between cholera toxin, cholera toxin substrates, a membrane protein S that is required for toxin activity, and a soluble protein CF that is needed for the function of S. The toxin has little intrinsic ability to catalyze ADP-ribosylations unless it encounters the active form of the S protein, which is S liganded to GTP or to a GTP analogue. In the presence of CF, S x GTP forms readily, though reversibly, but a more permanent active species, S-guanosine 5'-O-(3-thiotriphosphate) (S x GTP..gamma..S), forms over a period of 10-15 min at 37/sup 0/C. Both guanosine 5'-O-(2-thiodiphosphate) and GTP block this quasi-permanent activation. Some S x GTP..gamma..S forms in membranes that are exposed to CF alone and then to GTP..gamma..S, with a wash in between, and it is possible that CF facilitates a G nucleotide exchange. S x GTP..gamma..S dissolved by nonionic detergents persists in solution and can be used to support the ADP-ribosylation of nucleotide-free substrates. In this circumstance, added guanyl nucleotides have no further effect. This active form of S is unstable, especially when heated, but the thermal inactivation above 45/sup 0/C is decreased by GTP..gamma..S. Active S is required equally for the ADP-ribosylation of all of cholera toxin's protein substrates, regardless of whether they bind GTP or not. They suggest that active S interacts directly with the enzymic A/sub 1/ fragments of cholera toxin and not with any toxin substrate. The activation and activity of S are independent of the state, or even the presence, of adenylate cyclase and seem to be involved with the cyclase system only via cholera toxin. S is apparently not related by function to certain other GTP binding proteins, including p21/sup ras/, and appears to be a new GTP binding protein whose physiologic role remains to be identified.

  10. A biomimetic nanosponge that absorbs pore-forming toxins

    NASA Astrophysics Data System (ADS)

    Hu, Che-Ming J.; Fang, Ronnie H.; Copp, Jonathan; Luk, Brian T.; Zhang, Liangfang

    2013-05-01

    Detoxification treatments such as toxin-targeted anti-virulence therapy offer ways to cleanse the body of virulence factors that are caused by bacterial infections, venomous injuries and biological weaponry. Because existing detoxification platforms such as antisera, monoclonal antibodies, small-molecule inhibitors and molecularly imprinted polymers act by targeting the molecular structures of toxins, customized treatments are required for different diseases. Here, we show a biomimetic toxin nanosponge that functions as a toxin decoy in vivo. The nanosponge, which consists of a polymeric nanoparticle core surrounded by red blood cell membranes, absorbs membrane-damaging toxins and diverts them away from their cellular targets. In a mouse model, the nanosponges markedly reduce the toxicity of staphylococcal alpha-haemolysin (?-toxin) and thus improve the survival rate of toxin-challenged mice. This biologically inspired toxin nanosponge presents a detoxification treatment that can potentially treat a variety of injuries and diseases caused by pore-forming toxins.

  11. Clostridium perfringens Delta-Toxin Induces Rapid Cell Necrosis

    PubMed Central

    Seike, Soshi; Miyamoto, Kazuaki; Kobayashi, Keiko; Takehara, Masaya; Nagahama, Masahiro

    2016-01-01

    Clostridium perfringens delta-toxin is a β-pore-forming toxin and a putative pathogenic agent of C. perfringens types B and C. However, the mechanism of cytotoxicity of delta-toxin remains unclear. Here, we investigated the mechanisms of cell death induced by delta-toxin in five cell lines (A549, A431, MDCK, Vero, and Caco-2). All cell lines were susceptible to delta-toxin. The toxin caused rapid ATP depletion and swelling of the cells. Delta-toxin bound and formed oligomers predominantly in plasma membrane lipid rafts. Destruction of the lipid rafts with methyl β-cyclodextrin inhibited delta-toxin-induced cytotoxicity and ATP depletion. Delta-toxin caused the release of carboxyfluorescein from sphingomyelin-cholesterol liposomes and formed oligomers; toxin binding to the liposomes declined with decreasing cholesterol content in the liposomes. Flow cytometric assays with annexin V and propidium iodide revealed that delta-toxin treatment induced an elevation in the population of annexin V-negative and propidium iodide-positive cells. Delta-toxin did not cause the fragmentation of DNA or caspase-3 activation. Furthermore, delta-toxin caused damage to mitochondrial membrane permeability and cytochrome c release. In the present study, we demonstrate that delta-toxin produces cytotoxic activity through necrosis. PMID:26807591

  12. In situ high-resolution evaluation of labile arsenic and mercury in sediment of a large shallow lake.

    PubMed

    Wang, Chao; Yao, Yu; Wang, Peifang; Hou, Jun; Qian, Jin; Yuan, Ye; Fan, Xiulei

    2016-01-15

    The precise evaluation of arsenic (As) and mercury (Hg) bioavailability in sediment is crucial to controlling As and Hg contamination, but traditional ex situ measurements hamper comprehensive analysis of labile As and Hg in sediment. In this study, we characterized in situ labile As and Hg in sediment of Lake Hongze using the zirconium (Zr) oxide diffusive gradients in thin films (DGT) technique and 3-mercaptopropyl functionalized silica gel DGT, respectively. The concentrations of DGT-labile As and Hg in the sediment profiles were found to exhibit considerable variation, ranging from 0.15 to 4.15?gL(-1) for As and from 0.04 to 1.35?gL(-1) for Hg. As and Hg flux values, calculated based on the concentration gradients measured from the DGT profiles for both the overlying water and sediment close to the sediment-water interface, were used to determine the contamination status of As and Hg. Flux values of As and Hg were between -0.066 and 0.067ngcm(-2)d(-1) and between -0.0187 and 0.0181ngcm(-2)d(-1), respectively. The GNU's Not Unix R (GNU R) programming language was used to identify outliers of As and Hg at various depths at the sampling sites. The results indicate that the sites with the most outliers were all located in the regions that were seriously affected by contaminants from the Huai River. The DGT-labile As and Hg concentrations in the 0-30mm layer were found to be significantly correlated with concentrations of labile As and Hg, total dissolved As and Hg, and total As and Hg in the overlying water, as indicated by ex situ measurements. Results show that DGT is a reliable and high-resolution technique that can be used for in situ monitoring of the labile fractions of As and Hg in sediment in fresh water bodies. PMID:26398454

  13. Synthesis and biology of cyclic imine toxins, an emerging class of potent, globally distributed marine toxins.

    PubMed

    Stivala, Craig E; Benoit, Evelyne; Aroz, Rmulo; Servent, Denis; Novikov, Alexei; Molg, Jordi; Zakarian, Armen

    2015-03-01

    From a small group of exotic compounds isolated only two decades ago, Cyclic Imine (CI) toxins have become a major class of marine toxins with global distribution. Their distinct chemical structure, biological mechanism of action, and intricate chemistry ensures that CI toxins will continue to be the subject of fascinating fundamental studies in the broad fields of chemistry, chemical biology, and toxicology. The worldwide occurrence of potent CI toxins in marine environments, their accumulation in shellfish, and chemical stability are important considerations in assessing risk factors for human health. This review article aims to provide an account of chemistry, biology, and toxicology of CI toxins from their discovery to the present day. PMID:25338021

  14. Synthesis and Biology of Cyclic Imine Toxins, An Emerging Class of Potent, Globally Distributed Marine Toxins

    PubMed Central

    Stivala, Craig E.; Benoit, Evelyne; Araoz, Romulo; Servent, Denis; Novikov, Alexei

    2014-01-01

    From a small group of exotic compounds isolated only two decades ago, Cyclic Imine (CI) toxins have become a major class of marine toxins with global distribution. Their distinct chemical structure, biological mechanism of action, and intricate chemistry ensures that CI toxins will continue to be the subject of fascinating fundamental studies in the broad fields of chemistry, chemical biology, and toxicology. The worldwide occurrence of potent CI toxins in marine environments, their accumulation in shellfish, and chemical stability are important considerations in assessing risk factors for human health. This review article aims to provide an account of chemistry, biology, and toxicology of CI toxins from their discovery to the present day. PMID:25338021

  15. Shiga Toxin (Stx) Classification, Structure, and Function

    PubMed Central

    Melton-Celsa, Angela R.

    2014-01-01

    Shiga toxin (Stx) is one of the most potent bacterial toxins known. Stx is found in Shigella dysenteriae 1 and in some serogroups of Escherichia coli (called Stx1 in E. coli). In addition to or instead of Stx1, some E. coli strains produce a second type of Stx, Stx2, that has the same mode of action as Stx/Stx1 but that is antigenically distinct. Because subtypes of each toxin have been identified, the prototype toxin for each group is now designated Stx1a or Stx2a. The Stxs consist of two major subunits, an A subunit that joins noncovalently to a pentamer of five identical B subunits. The A subunit of the toxin injures the eukaryotic ribosome, and halts protein synthesis in target cells. The function of the B pentamer is to bind to the cellular receptor, globotriaosylceramide, Gb3, found primarily on endothelial cells. The Stxs traffic in a retrograde manner within the cell, such that the A subunit of the toxin reaches the cytosol only after the toxin moves from the endosome to the Golgi and then to the endoplasmic reticulum. In humans infected with Stx-producing E. coli (STEC), the most serious manifestation of the disease, the hemolytic uremic syndrome or HUS, is more often associated with strains that produce Stx2a rather than Stx1a, and that relative toxicity is replicated in mice and baboons. Stx1a and Stx2a also exhibit differences in cytotoxicity to various cell types, bind dissimilarly to receptor analogs or mimics, induce differential chemokine responses, and have several distinctive structural characteristics. PMID:25530917

  16. 14C as a tracer of labile organic matter in Antarctic benthic food webs

    NASA Astrophysics Data System (ADS)

    Purinton, Brett L.; DeMaster, David J.; Thomas, Carrie J.; Smith, Craig R.

    2008-11-01

    14C measurements were made on surface plankton, particle-trap material, surface sediment, benthic invertebrate gut contents, and body tissue samples to assess the effectiveness of this radioisotope as a tracer of labile organic carbon in Antarctic benthic food webs. Samples were collected on five cruises to the West Antarctic Peninsula (WAP) shelf between November 1999 and March 2001 as part of the Food for Benthos on the ANtarctic Continental-Shelf (FOODBANCS) Project. The 14C contents of the body tissues from a variety of deposit feeders (-12613 per mil) were substantially enriched relative to the surface sediment (-23413 per mil) and statistically similar to the organic matter collected in plankton tows (-13510 per mil), indicating that recently produced marine plankton are the primary source of nutrition for these deposit feeders on the West Antarctic shelf. Selective ingestion was the primary feeding strategy used by echiuran worms and certain holothurians (i.e. Peniagone vignoni) for incorporating labile organic carbon into their tissues as demonstrated by the large differences (10513 per mil) between surface sediment and gut content 14C activities. In contrast, digestive and/or assimilatory selection was the predominant strategy used by an irregular urchin ( Amphipneustes lorioli) and several other holothurians ( Protelpidia murrayi, Bathyplotes fuscivinculum and the head-down conveyor belt feeder, Molpadia musculus), as demonstrated by large differences (427 per mil) between the 14C activities of their foregut or whole-gut organic contents and their body tissues. Despite large fluctuations in carbon export from the euphotic zone, benthic feeding strategies remained essentially constant over the 15-month sampling period. No seasonal variation was evident in either the 14C abundance of the deposit-feeder body tissues, or in the 14C abundance of their gut contents. The mean 14C abundance in the body tissues of the two sub-surface deposit feeders ( A. lorioli and M. musculus; mean=-136.28.5 per mil) was distinct ( p=0.0008) from the mean 14C abundance in the body tissues of the four surface deposit feeders (echiuran worm, P. vignoni, P. murrayi, and B. fusciviculum; -122.612.3 per mil). The mean 14C abundance of the gut contents from the sub-surface deposit feeders (-178.018.6 per mil) also was significantly depleted ( p=0.0009) relative to that of the surface deposit feeders (-149.526.6 per mil). The 14C measurements proved to be a much more sensitive tracer for tracking labile organic carbon during ingestive and assimilatory processes than the stable isotopes of carbon or nitrogen.

  17. The evolution of dioecy, heterodichogamy, and labile sex expression in Acer.

    PubMed

    Renner, S S; Beenken, L; Grimm, G W; Kocyan, A; Ricklefs, R E

    2007-11-01

    The northern hemisphere tree genus Acer comprises 124 species, most of them monoecious, but 13 dioecious. The monoecious species flower dichogamously, duodichogamously (male, female, male), or in some species heterodichogamously (two morphs that each produce male and female flowers but at reciprocal times). Dioecious species cannot engage in these temporal strategies. Using a phylogeny for 66 species and subspecies obtained from 6600 nucleotides of chloroplast introns, spacers, and a protein-coding gene, we address the hypothesis (Pannell and Verdú, Evolution 60: 660-673. 2006) that dioecy evolved from heterodichogamy. This hypothesis was based on phylogenetic analyses (Gleiser and Verdú, New Phytol. 165: 633-640. 2005) that included 29-39 species of Acer coded for five sexual strategies (duodichogamous monoecy, heterodichogamous androdioecy, heterodichogamous trioecy, dichogamous subdioecy, and dioecy) treated as ordered states or as a single continuous variable. When reviewing the basis for these scorings, we found errors that together with the small taxon sample, cast doubt on the earlier inferences. Based on published studies, we coded 56 species of Acer for four sexual strategies, dioecy, monoecy with dichogamous or duodichogamous flowering, monoecy with heterodichogamous flowering, or labile sex expression, in which individuals reverse their sex allocation depending on environment-phenotype interactions. Using Bayesian character mapping, we infer an average of 15 transformations, a third of them involving changes from monoecy-cum-duodichogamy to dioecy; less frequent were changes from this strategy to heterodichogamy; dioecy rarely reverts to other sexual systems. Contra the earlier inferences, we found no switches between heterodichogamy and dioecy. Unexpectedly, most of the species with labile sex expression are grouped together, suggesting that phenotypic plasticity in Acer may be a heritable sexual strategy. Because of the complex flowering phenologies, however, a concern remains that monoecy in Acer might not always be distinguishable from labile sex expression, which needs to be addressed by long-term monitoring of monoecious trees. The 13 dioecious species occur in phylogenetically disparate clades that date back to the Late Eocene and Oligocene, judging from a fossil-calibrated relaxed molecular clock. PMID:17894810

  18. Structural insights into the mechanism of activation of the TRPV1 channel by a membrane-bound tarantula toxin.

    PubMed

    Bae, Chanhyung; Anselmi, Claudio; Kalia, Jeet; Jara-Oseguera, Andres; Schwieters, Charles D; Krepkiy, Dmitriy; Won Lee, Chul; Kim, Eun-Hee; Kim, Jae Il; Faraldo-Gmez, Jos D; Swartz, Kenton J

    2016-01-01

    Venom toxins are invaluable tools for exploring the structure and mechanisms of ion channels. Here, we solve the structure of double-knot toxin (DkTx), a tarantula toxin that activates the heat-activated TRPV1 channel. We also provide improved structures of TRPV1 with and without the toxin bound, and investigate the interactions of DkTx with the channel and membranes. We find that DkTx binds to the outer edge of the external pore of TRPV1 in a counterclockwise configuration, using a limited protein-protein interface and inserting hydrophobic residues into the bilayer. We also show that DkTx partitions naturally into membranes, with the two lobes exhibiting opposing energetics for membrane partitioning and channel activation. Finally, we find that the toxin disrupts a cluster of hydrophobic residues behind the selectivity filter that are critical for channel activation. Collectively, our findings reveal a novel mode of toxin-channel recognition that has important implications for the mechanism of thermosensation. PMID:26880553

  19. Structural insights into the mechanism of activation of the TRPV1 channel by a membrane-bound tarantula toxin

    PubMed Central

    Bae, Chanhyung; Anselmi, Claudio; Kalia, Jeet; Jara-Oseguera, Andres; Schwieters, Charles D; Krepkiy, Dmitriy; Won Lee, Chul; Kim, Eun-Hee; Kim, Jae Il; Faraldo-Gmez, Jos D; Swartz, Kenton J

    2016-01-01

    Venom toxins are invaluable tools for exploring the structure and mechanisms of ion channels. Here, we solve the structure of double-knot toxin (DkTx), a tarantula toxin that activates the heat-activated TRPV1 channel. We also provide improved structures of TRPV1 with and without the toxin bound, and investigate the interactions of DkTx with the channel and membranes. We find that DkTx binds to the outer edge of the external pore of TRPV1 in a counterclockwise configuration, using a limited protein-protein interface and inserting hydrophobic residues into the bilayer. We also show that DkTx partitions naturally into membranes, with the two lobes exhibiting opposing energetics for membrane partitioning and channel activation. Finally, we find that the toxin disrupts a cluster of hydrophobic residues behind the selectivity filter that are critical for channel activation. Collectively, our findings reveal a novel mode of toxin-channel recognition that has important implications for the mechanism of thermosensation. DOI: http://dx.doi.org/10.7554/eLife.11273.001 PMID:26880553

  20. Cyanobacterial Toxin Degrading Bacteria: Who Are They?

    PubMed Central

    Kormas, Konstantinos Ar.; Lymperopoulou, Despoina S.

    2013-01-01

    Cyanobacteria are ubiquitous in nature and are both beneficial and detrimental to humans. Benefits include being food supplements and producing bioactive compounds, like antimicrobial and anticancer substances, while their detrimental effects are evident by toxin production, causing major ecological problems at the ecosystem level. To date, there are several ways to degrade or transform these toxins by chemical methods, while the biodegradation of these compounds is understudied. In this paper, we present a meta-analysis of the currently available 16S rRNA and mlrA (microcystinase) genes diversity of isolates known to degrade cyanobacterial toxins. The available data revealed that these bacteria belong primarily to the Proteobacteria, with several strains from the sphingomonads, and one from each of the Methylobacillus and Paucibacter genera. Other strains belonged to the genera Arthrobacter, Bacillus, and Lactobacillus. By combining the ecological knowledge on the distribution, abundance, and ecophysiology of the bacteria that cooccur with toxic cyanobacterial blooms and newly developed molecular approaches, it is possible not only to discover more strains with cyanobacterial toxin degradation abilities, but also to reveal the genes associated with the degradation of these toxins. PMID:23841072

  1. Array biosensor for detection of toxins

    NASA Technical Reports Server (NTRS)

    Ligler, Frances S.; Taitt, Chris Rowe; Shriver-Lake, Lisa C.; Sapsford, Kim E.; Shubin, Yura; Golden, Joel P.

    2003-01-01

    The array biosensor is capable of detecting multiple targets rapidly and simultaneously on the surface of a single waveguide. Sandwich and competitive fluoroimmunoassays have been developed to detect high and low molecular weight toxins, respectively, in complex samples. Recognition molecules (usually antibodies) were first immobilized in specific locations on the waveguide and the resultant patterned array was used to interrogate up to 12 different samples for the presence of multiple different analytes. Upon binding of a fluorescent analyte or fluorescent immunocomplex, the pattern of fluorescent spots was detected using a CCD camera. Automated image analysis was used to determine a mean fluorescence value for each assay spot and to subtract the local background signal. The location of the spot and its mean fluorescence value were used to determine the toxin identity and concentration. Toxins were measured in clinical fluids, environmental samples and foods, with minimal sample preparation. Results are shown for rapid analyses of staphylococcal enterotoxin B, ricin, cholera toxin, botulinum toxoids, trinitrotoluene, and the mycotoxin fumonisin. Toxins were detected at levels as low as 0.5 ng mL(-1).

  2. Therapy and prophylaxis of inhaled biological toxins.

    PubMed

    Paddle, Brian M

    2003-01-01

    This review highlights the current lack of therapeutic and prophylactic treatments for use against inhaled biological toxins, especially those considered as potential biological warfare (BW) or terrorist threats. Although vaccine development remains a priority, the use of rapidly deployable adjunctive therapeutic or prophylactic drugs could be life-saving in severe cases of intoxication or where vaccination has not been possible or immunity not established. The current lack of such drugs is due to many factors. Thus, methods involving molecular modelling are limited by the extent to which the cellular receptor sites and mode of action and structure of a toxin need to be known. There is also our general lack of knowledge of what effect individual toxins will have when inhaled into the lungs - whether and to what extent the action will be cell specific and cytotoxic or rather an acute inflammatory response requiring the use of immunomodulators. Possible sources of specific high-affinity toxin antagonists being investigated include monoclonal antibodies, selected oligonucleotides (aptamers) and derivatized dendritic polymers (dendrimers). The initial selection of suitable agents of these kinds can be made using cytotoxicity assays involving cultured normal human lung cells and a range of suitable indicators. The possibility that a mixture of selected antibody, aptamer or dendrimer-based materials for one or more toxins could be delivered simultaneously as injections or as inhaled aerosol sprays should be investigated. PMID:12794937

  3. Interaction of diphtheria toxin with phosphorylated molecules.

    PubMed Central

    Proia, R L; Hart, D A; Eidels, L

    1979-01-01

    The binding of diphtheria toxin to 125I-labeled cell surface glycoproteins from hamster thymocytes was shown to be inhibited by nucleotides. The relative effectiveness of the nucleotides (at 5 mM) was found to be thymidine triphosphate greater than adenosine triphosphate greater than guanosine triphosphate greater than uridine triphosphate greater than cytidine triphosphate. When adenine-containing compounds were used, the relative effectiveness was determined to be adenosine tetraphosphate greater than adenosine triphosphate greater than adenosine diphosphate greater than adenosine monophosphate. In addition, tetrapolyphosphate, tripolyphosphate, inositol hexaphosphate (phytic acid), and the highly phosphorylated proteins casein and phosvitin were also shown to be potent inhibitors of the binding of diphtheria toxin to 125I-labeled cell surface glycoproteins. Diphtheria toxin was shown to bind directly to 125I-casein; this binding was also inhibited by the highly phosphorylated compounds and was decreased by pretreatment of the 125I-casein with alkaline phosphatase. These results suggest that diphtheria toxin binds to regions of high phosphate density and raise the possibility that the site on the cell surface glycoproteins to which diphtheria toxin binds might be polyanionic in nature. PMID:528059

  4. Structure and function of insulin-like growth factor acid-labile subunits in mammalian homologues.

    PubMed

    Guilin, Li; Lili, Niu; Haifeng, Liu; Jiazhong, Guo

    2015-12-01

    Insulin-like growth factors (IGFs) act as a critical signaling pathway in animals and play significant roles in body growth, development, and occurrence and progression of animal diseases. In the past, structural and functional studies of ligands, receptors and even specific binding proteins in the IGFs system have been extensively investigated. However, the functional study of insulin-like growth factor acid-labile subunit (IGFALS) mainly focused on the prolonging half-life of IGFs. Increasing number of studies indicated that mutations in the IGFALS DNA sequence and low expression level of IGFALS proteins can lead to growth and development retardation in animals. In this review, we summarize recent structural and functional studies of IGFALS in mammals, aiming to further identify detailed genetic mechanism of IGFALS. PMID:26704943

  5. A superior drug carrier – aponeocarzinostatin in partially unfolded state fully protects the labile antitumor enediyne

    PubMed Central

    Shanmuganathan, Aranganathan; Kumar, Thallapuranam Krishnaswamy Suresh; Huang, Chiy-Mey; Yu, Chin; Chin, Der-Hang

    2009-01-01

    Background Neocarzinostatin is a potent antitumor drug consisting of an enediyne chromophore and a protein carrier. Methods We characterized an intermediate in the equilibrium unfolding pathway of aponeocarzinostatin, using a variety of biophysical techniques including 1-anilino-8-napthalene sulfonate binding studies, size-exclusion fast protein liquid chromatography, intrinsic tryptophan fluorescence, circular dichroism, and 1H-15N heteronuclear single quantum coherence spectroscopy. Results The partially unfolded protein is in molten globule-like state, in which ~60% and ~20% tertiary and secondary structure is disrupted respectively. Despite lacking a fully coordinated tertiary structure for assembling a functional binding cleft, the protein in molten globule-like state is still able to fully protect the labile chromophore. Titration of chromophore leads the partially denatured apoprotein to fold into its native state. Conclusion These findings bring insight into conserving mechanism of neocarzinostatin under harsh environment, where even the partially denatured apoprotein exhibits protective effect, confirming the superiority of the drug carrier. PMID:19463188

  6. Significance of Isotopically Labile Organic Hydrogen in Thermal Maturation of Organic Matter

    SciTech Connect

    Arndt Schimmelmann; Maria Mastalerz

    2010-03-30

    Isotopically labile organic hydrogen in fossil fuels occupies chemical positions that participate in isotopic exchange and in chemical reactions during thermal maturation from kerogen to bitumen, oil and gas. Carbon-bound organic hydrogen is isotopically far less exchangeable than hydrogen bound to nitrogen, oxygen, or sulfur. We explore why organic hydrogen isotope ratios express a relationship with organic nitrogen isotope ratios in kerogen at low to moderate maturity. We develop and apply new techniques to utilize organic D/H ratios in organic matter fractions and on a molecular level as tools for exploration for fossil fuels and for paleoenvironmental research. The scope of our samples includes naturally and artificially matured substrates, such as coal, shale, oil and gas.

  7. Genetic characteristics of toxigenic Clostridia and toxin gene evolution.

    PubMed

    Popoff, Michel R; Bouvet, Philippe

    2013-12-01

    Clostridia comprise a heterogenous group of environmental bacteria containing 15 pathogenic species, which produce the most potent toxins. The origin of toxins is still enigmatic. It is hypothesized that toxins exhibiting an enzymatic activity have derived from hydrolytic enzymes, which are abundantly secreted by these bacteria, and that pore-forming toxins have evolved from an ancestor transmembrane protein. The presence of related toxin genes in distinct Clostridium species and the variability of some toxin genes support horizontal toxin gene transfer and subsequent independent evolution from strain to strain. Clostridium perfringens toxin genes involved in myonecrosis, mainly alpha toxin and perfringolysin genes, are chromosomally located, whereas toxin genes responsible for intestinal and food borne diseases are localized on plasmids except the enterotoxin gene which can be located either on the chromosome or plasmids. The distribution of these plasmids containing one or several toxin genes accounts for the diverse C. perfringens toxinotypes. Clostridium difficile strains show a high genetic variability. But in contrast to C. perfringens, toxin genes are clustered in pathogenicity locus located on chromosome. The presence of related toxin genes in distinct clostridial species like Clostridium sordellii, Clostridium novyi, and C. perfringens supports interspecies mobilization of this locus. The multiple C. difficile toxinotypes based on toxin gene variants possibly reflect strain adaptation to the intestinal environment. Botulinum toxin genes also show a high level of genetic variation. They have a diverse genetic localization including chromosome, plasmid or phage, and are spread in various Clostridium species (Clostridium botulinum groups, Clostridium argentinense, Clostridium butyricum, Clostridium baratii). Exchange of toxin genes not only include transfers between Clostridium species but also between Clostridium and other bacterial species as well as eukaryotic cells as supported by the wide distribution of related pore-forming toxins of the aerolysin family in various clostridial and non-clostridial species, animal, mushroom and plant. PMID:23707611

  8. Labile and recalcitrant organic matter utilization by river biofilm under increasing water temperature.

    PubMed

    Ylla, Irene; Roman, Anna M; Sabater, Sergi

    2012-10-01

    Microbial biofilms in rivers contribute to the decomposition of the available organic matter which typically shows changes in composition and bioavailability due to their origin, seasonality, and watershed characteristics. In the context of global warming, enhanced biofilm organic matter decomposition would be expected but this effect could be specific when either a labile or a recalcitrant organic matter source would be available. A laboratory experiment was performed to mimic the effect of the predicted increase in river water temperature (+4C above an ambient temperature) on the microbial biofilm under differential organic matter sources. The biofilm microbial community responded to higher water temperature by increasing bacterial cell number, respiratory activity (electron transport system) and microbial extracellular enzymes (extracellular enzyme activity). At higher temperature, the phenol oxidase enzyme explained a large fraction of respiratory activity variation suggesting an enhanced microbial use of degradation products from humic substances. The decomposition of hemicellulose (?-xylosidase activity) seemed to be also favored by warmer conditions. However, at ambient temperature, the enzymes highly responsible for respiration activity variation were ?-glucosidase and leu-aminopeptidase, suggesting an enhanced microbial use of polysaccharides and peptides degradation products. The addition of labile dissolved organic carbon (DOC; dipeptide plus cellobiose) caused a further augmentation of heterotrophic biomass and respiratory activity. The changes in the fluorescence index and the ratio Abs(250)/total DOC indicated that higher temperature accelerated the rates of DOC degradation. The experiment showed that the more bioavailable organic matter was rapidly cycled irrespective of higher temperature while degradation of recalcitrant substances was enhanced by warming. Thus, pulses of carbon at higher water temperature might have consequences for DOC processing. PMID:22570120

  9. Phenotypic lability and the evolution of predator-induced plasticity in tadpoles.

    PubMed

    Van Buskirk, J

    2002-02-01

    The hypothesis that predator-induced defenses in anuran larvae are maintained by divergent selection across multiple predation environments has not been fully supported by empirical results. One reason may be that traits that respond slowly to environmental variation experience a fitness cost not incorporated in the standard adaptive model, due to a time lag between detecting the state of the environment and expressing the phenotypic response. I measured the rate at which behavior and morphology of Rana temporaria tadpoles change when confronted with a switch in the predation environment at two points in development. Hatchling tadpoles that had been exposed during the egg stage to Aeshna dragonfly larvae were not phenotypically different from those exposed as eggs to predator-free conditions, and both responded similarly to post-hatching predator treatments. When 25-day-old tadpoles from treatments with and without dragonflies were subjected to a switch in the environment, their activity budgets reversed completely within 24-36 h, and their body and tail shape began changing significantly within 4 days. The behavioral response was conservative: Tadpoles switched from high-risk to predator-free treatments were slower to adjust their activity. The study confirmed that behavioral traits are relatively labile and exhibit strong plasticity, but it did not reveal such a pattern at the level of individual traits: Morphological traits that developed slowly did not show the least plasticity. Thus, I found that differences in lability of traits were useful for predicting the magnitude of plasticity only for fundamentally different kinds of characters. PMID:11926504

  10. Effect of temperature on the decomposition rate of labile and stable organic matter in an agrochernozem

    NASA Astrophysics Data System (ADS)

    Larionova, A. A.; Kvitkina, A. K.; Yevdokimov, I. V.; Bykhovets, S. S.; Stulin, A. F.

    2014-05-01

    An hypothesis about the different temperature dependences of the decomposition of the labile and stable organic carbon pools has been tested using an agrochernozem sampled from an experimental plot of 42-year-old continuous corn in Voronezh oblast. The partitioning of the CO2 loss during the decomposition of the labile and stable soil organic matter (SOM) at 2, 12, and 22C in a long-term incubation experiment was performed using the method of 13C natural abundance by C3-C4 transition. On the basis of the determined decomposition constants, the SOM pools have been arranged in an order according to their increasing stability: plant residues < new (C4) SOM < old (C3) SOM. The tested hypothesis has been found valid only for a limited temperature interval. The temperature coefficient Q 10 increases in the stability order from 1.2 to 4.3 in the interval of 12-22C. At low temperatures (2-12C), the values of Q 10 insignificantly vary among the SOM pools and lie in the range of 2.2-2.8. Along with the decomposition constants of the SOM, the new-to-old carbon ratio in the CO2 efflux from the soil and the magnitude of the negative priming effect for the old SOM caused by the input of new organic matter depend on the temperature. In the soil under continuous corn fertilized with NPK, the increased decomposition of C3 SOM is observed compared to the unfertilized control; the temperature dependences of the SOM decomposition are similar in both agrochernozem treatments.

  11. Following The Money: Characterizing the Dynamics of Microbial Ecosystems and Labile Organic Matter in Grassland Soils

    NASA Astrophysics Data System (ADS)

    Herbert, B. E.; McNeal, K. S.

    2006-12-01

    The dynamics of soil microbial ecosystems and labile fractions of soil organic matter in grasslands have important implications for the response of these critical ecosystems to perturbations. Organic, inorganic and genetic biomarkers in the solid (e.g. lipids, microbial DNA), liquid (e.g. porewater ions) or gaseous phases (e.g. carbon dioxide) have been used to characterize carbon cycling and soil microbial ecology. These proxies are generally limited in the amount of temporal information that they can provide (i.e., solid-phase proxies) or the amount of specific information they can provide about carbon sources or microbial community processes (e.g. inorganic gases). It is the aim of this research to validate the use of soil volatile organic carbon emissions (VOCs) as useful indicators of subsurface microbial community shifts and processes as a function of ecosystem perturbations. We present results of method validation using laboratory microcosm, where VOC metabolites as characterized by gas chromatography and mass spectrometry (GC-MS), were related to other proxies including carbon dioxide (CO2) via infra-red technology, and microbial community shifts as measured by Biolog and fatty acid methyl ester (FAME) techniques. Experiments with soil collected from grasslands along the coastal margin region in southern Texas were preformed where environmental factors such as soil water content, soil type, and charcoal content are manipulated. Results indicate that over fifty identifiable VOC metabolites are produced from the soils, where many (~15) can be direct indicators of microbial ecology. Principle component analysis (PCA) evidences these trends through similar cluster patterns for the VOC results, the Biolog results, and FAME. Regression analysis further shows that VOCs are significant (p < 0.05) indicators of microbial stress. Our results are encouraging that characterizing VOCs production in grassland soils are easy to measure, relatively inexpensive method, and useful proxies of subsurface microbial ecosystems and the dynamics of labile carbon in these systems.

  12. In vivo lability of glucose-6-phosphate dehydrogenase in GdA- and Gdmediterranean deficiency

    PubMed Central

    Piomelli, Sergio; Corash, Laurence M.; Davenport, Deatra D.; Miraglia, Janet; Amorosi, Edward L.

    1968-01-01

    A decreased level of glucose-6-phosphate dehydrogenase might result from decreased rate of synthesis, synthesis of an enzyme of lower catalytic efficiency, increased lability, or a combined mechanism. To test the hypothesis of increased lability, the rate of decline of the enzyme in vivo was measured in three groups of individuals, controls, Gd(),A-males, and Gd(), Mediterranean males, by the slope of decline of activity in fractions containing erythrocytes of progressively increasing mean age. These fractions were obtained by ultracentrifugation on a discontinuous density gradient of erythrocyte suspensions free of contaminating platelets and leukocytes. The rate of in vivo decline of pyruvate kinase (another age-dependent enzyme) was also measured and found very similar in the three groups. The in vivo decline of glucose-6-phosphate dehydrogenase was found to follow an exponential rate, with a half-life of 62 days for controls and 13 days for Gd(),A- erythrocytes. The activity in normal reticulocytes was estimated at 9.7 U and in Gd(),A- reticulocytes at 8.8 U. These estimates were confirmed by direct measurements in reticulocytes isolated from patients with extreme reticulocytosis. In Gd(),Mediterranean erythrocytes activity could be demonstrated only in reticulocytes, which were estimated to average 1.4 U. The rate of decline is so extreme that no activity could be detected in mature erythrocytes. These data suggest that the glucose-6-phosphate dehydrogenase deficiency of both the GdA- and the GdMediterranean variant results from different degrees of in vivo instability of the abnormal enzyme. PMID:5641629

  13. High lability of sexual system over 250 million years of evolution in morphologically conservative tadpole shrimps

    PubMed Central

    2013-01-01

    Background Sexual system is a key factor affecting the genetic diversity, population structure, genome structure and the evolutionary potential of species. The sexual system androdioecy where males and hermaphrodites coexist in populations is extremely rare, yet is found in three crustacean groups, barnacles, a genus of clam shrimps Eulimnadia, and in the order Notostraca, the tadpole shrimps. In the ancient crustacean order Notostraca, high morphological conservatism contrasts with a wide diversity of sexual systems, including androdioecy. An understanding of the evolution of sexual systems in this group has been hampered by poor phylogenetic resolution and confounded by the widespread occurrence of cryptic species. Here we use a multigene supermatrix for 30 taxa to produce a comprehensive phylogenetic reconstruction of Notostraca. Based on this phylogenetic reconstruction we use character mapping techniques to investigate the evolution of sexual systems. We also tested the hypothesis that reproductive assurance has driven the evolution of androdioecy in Notostraca. Results Character mapping analysis showed that sexual system is an extremely flexible trait within Notostraca, with repeated shifts between gonochorism and androdioecy, the latter having evolved a minimum of five times. In agreement with the reproductive assurance hypothesis androdioecious notostracans are found at significantly higher latitudes than gonochoric ones indicating that post glacial re-colonisation may have selected for the higher colonisation ability conferred by androdioecy. Conclusions In contrast to their conserved morphology, sexual system in Notostraca is highly labile and the rare reproductive mode androdioecy has evolved repeatedly within the order. Furthermore, we conclude that this lability of sexual system has been maintained for at least 250 million years and may have contributed to the long term evolutionary persistence of Notostraca. Our results further our understanding of the evolution of androdioecy and indicate that reproductive assurance is a recurrent theme involved in the evolution of this sexual system. PMID:23384124

  14. Preliminary Reliability Study of the Affective Lability Scale Adapted for Adolescents in a Francophone Clinical Population

    PubMed Central

    Guil, Jean Marc; Chapdelaine, Cimon; Desrosiers, Lyne; Cornez, Christine; Bouvier, Hlne; Breton, Jean-Jacques

    2009-01-01

    Objective: to study the reliability of a self-questionnaire on adolescent affective instability, within a francophone clinical population. Method: After reviewing the literature and consulting experts, the Affective Lability Scale (ALS), developed by Harvey and collaborators (1989), was selected. An anglophone and francophone version, adapted for the adolescent population, was developed. The final version includes 54 questions, divided into six sections, which examine the affective variations between euthymia, on the one hand, and depression, elation, and anxiety, on the other, and also examine the affective variations between anxiety and depression, and between depression and elation. The francophone version was the subject of a reliability study. A total of 43 francophone adolescents (48.8% male, 51.2% female; median age=14.86 years) took part in the comprehension and reliability study. Results: All questions were assigned a comprehension rate per item (CRI)>0.60 as evaluated by inter-rater agreement (median kappa=0.85). The median CRI is very satisfactory (0.88, SD=0.14). The internal consistency, determined by Cronbach coefficients, is elevated for each section and for the entire instrument (0.87 to 0.95). The temporal stability at three weeks is satisfactory, with an intraclass correlation coefficient (ICC) for the entire scale of 0.89 (confidence interval of 95%=0.75 to 0.95) and sections ICCs ranging between 0.72 and 0.89. Conclusion: This initial reliability data supports interest in the ALS as a means to evaluate a global level of affective lability, during the euthymic phase, among francophone adolescents in an in-patient psychiatric unit. A future study with a larger sample will make it possible to confirm these initial results and examine the factorial structure of the instrument.

  15. Purification and characterization of the lethal toxin (alpha-toxin) of Clostridium septicum.

    PubMed

    Ballard, J; Bryant, A; Stevens, D; Tweten, R K

    1992-03-01

    Clostridium septicum lethal (alpha-toxin) was purified and found to be a basic protein (pI 8.4) of approximately 48 kDa that is both lethal and hemolytic. The alpha-toxin had a hemolytic activity of approximately 2 x 10(7) hemolytic units per mg and a 50% lethal dose of approximately 10 micrograms/kg of body weight for mice. The alpha-toxin formed concentration-dependent, sodium dodecyl sulfate-resistant aggregates of approximately 230 kDa. Mice immunized with alpha-toxin showed a significant increase in survival time over mock-immunized mice when challenged with C. septicum. Rabbit polyclonal antibody was generated against the purified toxin and was used to confirm that toxin with the same molecular weight was present in seven different C. septicum isolates. No proteins in the supernatants from cultures of Clostridium perfringens, Clostridium histolyticum, Clostridium chauvoei, or Clostridium difficile were found to react with the C. septicum alpha-toxin-specific antibody. PMID:1541552

  16. Purification and characterization of the lethal toxin (alpha-toxin) of Clostridium septicum.

    PubMed Central

    Ballard, J; Bryant, A; Stevens, D; Tweten, R K

    1992-01-01

    Clostridium septicum lethal (alpha-toxin) was purified and found to be a basic protein (pI 8.4) of approximately 48 kDa that is both lethal and hemolytic. The alpha-toxin had a hemolytic activity of approximately 2 x 10(7) hemolytic units per mg and a 50% lethal dose of approximately 10 micrograms/kg of body weight for mice. The alpha-toxin formed concentration-dependent, sodium dodecyl sulfate-resistant aggregates of approximately 230 kDa. Mice immunized with alpha-toxin showed a significant increase in survival time over mock-immunized mice when challenged with C. septicum. Rabbit polyclonal antibody was generated against the purified toxin and was used to confirm that toxin with the same molecular weight was present in seven different C. septicum isolates. No proteins in the supernatants from cultures of Clostridium perfringens, Clostridium histolyticum, Clostridium chauvoei, or Clostridium difficile were found to react with the C. septicum alpha-toxin-specific antibody. Images PMID:1541552

  17. Methods for the detection of marine toxins

    SciTech Connect

    Wekell, M.M.; Manger, R.M.; Hadley, S.W.; Hungerford, J.M.

    1995-12-01

    Toxic materials have been dumped into the seas from waste streams and other pollutant sources such as runoff, etc. For protection of public health, it is essential that rapid, reliable and simple methods exist to detect marine toxins in seafoods. In addition, it is necessary to develop methods requiring a minimum of test material. Pure standards for many of the marine toxins are scarce. Reduced sample requirements extend the utility of detection methods in research and forensic applications as well. In the past, there was much reliance on the animal bioassay; however, this dependence hopefully will be reduced as newer instrumental techniques (chromatographic, mass spectrometric, electrophoretic), biochemical (immunochemical, receptor site assay), and cell bioassay methods are developed with a higher degree of precision and specificity. It is beneficial that a multiplicity of methods be available to detect marine toxins in seafoods. Each method has unique advantages and disadvantages.

  18. Thermal Analysis of Labile Trace Elements in CM and CV Carbonaceous Chondrites Using Inductively Coupled Plasma-Mass Spectrometry

    NASA Technical Reports Server (NTRS)

    Lauretta, D. S.; Klaue, B.; Blum, J. D.; Buseck, P. R.

    2001-01-01

    We developed a technique to measure the thermal release profiles of a suite of labile elements (Zn, As, Se, Cd, In, Sn, Sb, Te, Pt, Hg, Au, Tl, Pb, Bi). Conclusions are reached about the behavior of each element during parent-body alteration. Additional information is contained in the original extended abstract.

  19. Plant-Soil Relationships of Bromus tectorum L.: Interactions among Labile Carbon Additions, Soil Invasion Status, and Fertilizer.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Invasion of western North America by the annual exotic grass Bromus tectorum L. (cheatgrass) has been an ecological disaster. High soil bioavailability of nitrogen is a contributing factor in the invasive potential of B. tectorum. Application of labile carbon sources to the soil can immobilize soil ...

  20. Estimating rates and patterns of morphological evolution from phylogenies: lessons in limb lability from Australian Lerista lizards

    PubMed Central

    Wiens, John J

    2009-01-01

    Squamates (lizards and snakes) offer an exciting model system for research on the evolution of body form. A new phylogenetic study in BMC Evolutionary Biology of Australian lizards shows remarkable evolutionary lability in digit numbers among closely related species, but also highlights important challenges in this area. PMID:19291259

  1. On the Labile Memory Buffer in the Attentional Blink: Masking the T2 Representation by Onset Transients Mediates the AB

    ERIC Educational Resources Information Center

    Jannati, Ali; Spalek, Thomas M.; Di Lollo, Vincent

    2011-01-01

    Report of a second target (T2) is impaired when presented within 500 ms of the first (T1). This attentional blink (AB) is known to cause a delay in T2 processing during which T2 must be stored in a labile memory buffer. We explored the buffer's characteristics using different types of masks after T2. These characteristics were inferred by…

  2. Emotional Lability in Children and Adolescents with Attention Deficit/Hyperactivity Disorder (ADHD): Clinical Correlates and Familial Prevalence

    ERIC Educational Resources Information Center

    Sobanski, Esther; Banaschewski, Tobias; Asherson, Philip; Buitelaar, Jan; Chen, Wai; Franke, Barbara; Holtmann, Martin; Krumm, Bertram; Sergeant, Joseph; Sonuga-Barke, Edmund; Stringaris, Argyris; Taylor, Eric; Anney, Richard; Ebstein, Richard P.; Gill, Michael; Miranda, Ana; Mulas, Fernando; Oades, Robert D.; Roeyers, Herbert; Rothenberger, Aribert; Steinhausen, Hans-Christoph; Faraone, Stephen V.

    2010-01-01

    Background: The goal of this study was to investigate the occurrence, severity and clinical correlates of emotional lability (EL) in children with attention deficit/hyperactivity disorder (ADHD), and to examine factors contributing to EL and familiality of EL in youth with ADHD. Methods: One thousand, one hundred and eighty-six children with ADHD

  3. A Longitudinal Study of Emotion Regulation, Emotion Lability-Negativity, and Internalizing Symptomatology in Maltreated and Nonmaltreated Children

    ERIC Educational Resources Information Center

    Kim-Spoon, Jungmeen; Cicchetti, Dante; Rogosch, Fred A.

    2013-01-01

    The longitudinal contributions of emotion regulation and emotion lability-negativity to internalizing symptomatology were examined in a low-income sample (171 maltreated and 151 nonmaltreated children, from age 7 to 10years). Latent difference score models indicated that for both maltreated and nonmaltreated children, emotion regulation was a

  4. On the Labile Memory Buffer in the Attentional Blink: Masking the T2 Representation by Onset Transients Mediates the AB

    ERIC Educational Resources Information Center

    Jannati, Ali; Spalek, Thomas M.; Di Lollo, Vincent

    2011-01-01

    Report of a second target (T2) is impaired when presented within 500 ms of the first (T1). This attentional blink (AB) is known to cause a delay in T2 processing during which T2 must be stored in a labile memory buffer. We explored the buffer's characteristics using different types of masks after T2. These characteristics were inferred by

  5. Emotional Lability in Children and Adolescents with Attention Deficit/Hyperactivity Disorder (ADHD): Clinical Correlates and Familial Prevalence

    ERIC Educational Resources Information Center

    Sobanski, Esther; Banaschewski, Tobias; Asherson, Philip; Buitelaar, Jan; Chen, Wai; Franke, Barbara; Holtmann, Martin; Krumm, Bertram; Sergeant, Joseph; Sonuga-Barke, Edmund; Stringaris, Argyris; Taylor, Eric; Anney, Richard; Ebstein, Richard P.; Gill, Michael; Miranda, Ana; Mulas, Fernando; Oades, Robert D.; Roeyers, Herbert; Rothenberger, Aribert; Steinhausen, Hans-Christoph; Faraone, Stephen V.

    2010-01-01

    Background: The goal of this study was to investigate the occurrence, severity and clinical correlates of emotional lability (EL) in children with attention deficit/hyperactivity disorder (ADHD), and to examine factors contributing to EL and familiality of EL in youth with ADHD. Methods: One thousand, one hundred and eighty-six children with ADHD…

  6. Tillage and rotational effects on exchangeable and enzyme-labile phosphorus forms in conventional and organic cropping systems

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The transformations of crop residues and bio-fertilizers used as primary sources of nutrients for organic grain and forage production are influenced by soil management practices. The effects of management of the near-surface zone on labile phosphorus (P) forms were studied in soil under three organ...

  7. Retrograde trafficking of AB5 toxins: mechanisms to therapeutics

    PubMed Central

    Mukhopadhyay, Somshuvra; Linstedt, Adam D.

    2013-01-01

    Bacterial AB5 toxins are a clinically relevant class of exotoxins that includes several well-known members such as Shiga, cholera and pertussis toxins. Infections with toxin-producing bacteria cause devastating human diseases that affect millions of individuals each year and have no definitive medical treatment. The molecular targets of AB5 toxins reside in the cytosol of infected cells, and the toxins reach the cytosol by trafficking through the retrograde membrane transport pathway that avoids degradative late endosomes and lysosomes. Focusing on Shiga toxin as the archetype member, we review recent advances in understanding the molecular mechanisms involved in the retrograde trafficking of AB5 toxins and highlight how these basic science advances are leading to the development of a promising new therapeutic approach based on inhibiting toxin transport. PMID:23665994

  8. On muscle spindles, dystonia and botulinum toxin.

    PubMed

    Rosales, R L; Dressler, D

    2010-07-01

    Dystonia may produce co-contractions and constant strain in numerous muscle fibers, including those of the muscle spindles. As proprioceptors, muscle spindles detect dynamic or static changes in muscle length and their afferent projections to the spinal cord play a central role in control of antagonistic muscles. Their parallel arrangement with extrafusal muscle fibers and association with the earlier recruited oxidative motor units allow them to conveniently sample the activity of all motor units and effectively modulate movement. At the same time, fusimotor muscle spindle innervation contracts the striated polar portions of the intrafusal muscle fibers and prevents their slackening during extrafusal muscle contractions. Botulinum toxin remains the most efficient therapy of dystonia. Its muscular mechanism of action is hinged on cholinergic blockade not only of extrafusal, but also of intrafusal muscle fibers. Besides being a targeted muscular therapy, the alteration of the corresponding sensory input following an effect of botulinum toxin on the intrafusal muscle fibers is pivotal in modulating loss of pre-synaptic inhibition in dystonia, including suppression of the tonic vibration reflex. Whether or not trans-synaptic botulinum toxin migration occurs, a modification of the central motor programming is bound to happen in dystonia, with botulinum toxin acting either as another 'sensory trick' or as a form of 'short-term plasticity'. Knowledge of the muscle spindle anatomy and function is key to unify our understanding of abnormal movements and of effects of botulinum toxin therapy. Thus, in dystonia, overactivity of muscles and increased spindle sensitivity are germane to botulinum toxin targets of action. PMID:20590812

  9. Marine Toxins Targeting Ion Channels

    PubMed Central

    Arias, Hugo R.

    2006-01-01

    This introductory minireview points out the importance of ion channels for cell communication. The basic concepts on the structure and function of ion channels triggered by membrane voltage changes, the so-called voltage-gated ion channels (VGICs), as well as those activated by neurotransmitters, the so-called ligand-gated ion channel (LGICs), are introduced. Among the most important VGIC superfamiles, we can name the voltage-gated Na+ (NaV), Ca2+ (CaV), and K+ (KV) channels. Among the most important LGIC super families, we can include the Cys-loop or nicotinicoid, the glutamate-activated (GluR), and the ATP-activated (P2XnR) receptor superfamilies. Ion channels are transmembrane proteins that allow the passage of different ions in a specific or unspecific manner. For instance, the activation of NaV, CaV, or KV channels opens a pore that is specific for Na+, Ca2+, or K+, respectively. On the other hand, the activation of certain LGICs such as nicotinic acetylcholine receptors, GluRs, and P2XnRs allows the passage of cations (e.g., Na+, K+, and/or Ca2+), whereas the activation of other LGICs such as type A ?-butyric acid and glycine receptors allows the passage of anions (e.g., Cl? and/or HCO3?). In this regard, the activation of NaV and CaV as well as ligand-gated cation channels produce membrane depolarization, which finally leads to stimulatory effects in the cell, whereas the activation of KV as well as ligand-gated anion channels induce membrane hyperpolarization that finally leads to inhibitory effects in the cell. The importance of these ion channel superfamilies is emphasized by considering their physiological functions throughout the body as well as their pathophysiological implicance in several neuronal diseases. In this regard, natural molecules, and especially marine toxins, can be potentially used as modulators (e.g., inhibitors or prolongers) of ion channel functions to treat or to alleviate a specific ion channel-linked disease (e.g., channelopaties).

  10. Bacterial Toxins and the Nervous System: Neurotoxins and Multipotential Toxins Interacting with Neuronal Cells

    PubMed Central

    Popoff, Michel R.; Poulain, Bernard

    2010-01-01

    Toxins are potent molecules used by various bacteria to interact with a host organism. Some of them specifically act on neuronal cells (clostridial neurotoxins) leading to characteristics neurological affections. But many other toxins are multifunctional and recognize a wider range of cell types including neuronal cells. Various enterotoxins interact with the enteric nervous system, for example by stimulating afferent neurons or inducing neurotransmitter release from enterochromaffin cells which result either in vomiting, in amplification of the diarrhea, or in intestinal inflammation process. Other toxins can pass the blood brain barrier and directly act on specific neurons. PMID:22069606

  11. Inhibiting bacterial toxins by channel blockage.

    PubMed

    Bezrukov, Sergey M; Nestorovich, Ekaterina M

    2016-03-01

    Emergent rational drug design techniques explore individual properties of target biomolecules, small and macromolecule drug candidates, and the physical forces governing their interactions. In this minireview, we focus on the single-molecule biophysical studies of channel-forming bacterial toxins that suggest new approaches for their inhibition. We discuss several examples of blockage of bacterial pore-forming and AB-type toxins by the tailor-made compounds. In the concluding remarks, the most effective rationally designed pore-blocking antitoxins are compared with the small-molecule inhibitors of ion-selective channels of neurophysiology. PMID:26656888

  12. The mechanisms of action of reproductive toxins.

    PubMed

    Mattison, D R

    1983-01-01

    A general discussion of the mechanisms of action of reproductive toxins is presented. Reproductive toxicants may produce their adverse effects by one of several mechanisms. Some xenobiotics may act directly, either by virtue of chemical reactivity (eg, oocyte destruction by alkylating agents) or by structural similarity to endogenous molecules (eg, hormone antagonists). Other xenobiotics interrupt reproductive processes indirectly either by metabolic processing to a direct-acting toxicant (eg, prohormone or metabolic activation to reactive intermediate) or by altering endocrine homeostasis (eg, increased steroid clearance). Selected examples of compounds demonstrating reproductive toxicity by one or more of these mechanisms are tabulated and an extensive bibliography of reproductive toxins is attached. PMID:6340485

  13. Use of botulinum toxin in dentistry.

    PubMed

    Hoque, Afreen; McAndrew, Maureen

    2009-11-01

    A growing number of dentists are providing botulinum toxin to patients. The research presented here outlines potential uses of Botox related to oral health and facial problems as compared to traditional treatment methods. The administration of Botox (historically done by dermatologists and neurologists) may fall under dentists' jurisdiction, as their training and knowledge encompasses the entire head and neck. A review is made of the literature, based on Ovid and PubMed searches, selecting articles describing the injection of botulinum toxin A in areas related to the oral cavity and the face, excluding cosmetic purposes. PMID:20069790

  14. Streptococcal toxins: role in pathogenesis and disease.

    PubMed

    Barnett, Timothy C; Cole, Jason N; Rivera-Hernandez, Tania; Henningham, Anna; Paton, James C; Nizet, Victor; Walker, Mark J

    2015-12-01

    Group A Streptococcus (Streptococcus pyogenes), group B Streptococcus (Streptococcus agalactiae) and Streptococcus pneumoniae (pneumococcus) are host-adapted bacterial pathogens among the leading infectious causes of human morbidity and mortality. These microbes and related members of the genus Streptococcus produce an array of toxins that act against human cells or tissues, resulting in impaired immune responses and subversion of host physiological processes to benefit the invading microorganism. This toxin repertoire includes haemolysins, proteases, superantigens and other agents that ultimately enhance colonization and survival within the host and promote dissemination of the pathogen. PMID:26433203

  15. Array Biosensor for Toxin Detection: Continued Advances

    PubMed Central

    Taitt, Chris Rowe; Shriver-Lake, Lisa C.; Ngundi, Miriam M.; Ligler, Frances S.

    2008-01-01

    The following review focuses on progress made in the last five years with the NRL Array Biosensor, a portable instrument for rapid and simultaneous detection of multiple targets. Since 2003, the Array Biosensor has been automated and miniaturized for operation at the point-of-use. The Array Biosensor has also been used to demonstrate (1) quantitative immunoassays against an expanded number of toxins and toxin indicators in food and clinical fluids, and (2) the efficacy of semi-selective molecules as alternative recognition moieties. Blind trials, with unknown samples in a variety of matrices, have demonstrated the versatility, sensitivity, and reliability of the automated system.

  16. Natural Toxins for Use in Pest Management

    PubMed Central

    Duke, Stephen O.; Cantrell, Charles L.; Meepagala, Kumudini M.; Wedge, David E.; Tabanca, Nurhayat; Schrader, Kevin K.

    2010-01-01

    Natural toxins are a source of new chemical classes of pesticides, as well as environmentally and toxicologically safer molecules than many of the currently used pesticides. Furthermore, they often have molecular target sites that are not exploited by currently marketed pesticides. There are highly successful products based on natural compounds in the major pesticide classes. These include the herbicide glufosinate (synthetic phosphinothricin), the spinosad insecticides, and the strobilurin fungicides. These and other examples of currently marketed natural product-based pesticides, as well as natural toxins that show promise as pesticides from our own research are discussed. PMID:22069667

  17. Shellfish toxicity: human health implications of marine algal toxins.

    PubMed

    James, K J; Carey, B; O'Halloran, J; van Pelt, F N A M; Skrabkov, Z

    2010-07-01

    Five major human toxic syndromes caused by the consumption of shellfish contaminated by algal toxins are presented. The increased risks to humans of shellfish toxicity from the prevalence of harmful algal blooms (HABs) may be a consequence of large-scale ecological changes from anthropogenic activities, especially increased eutrophication, marine transport and aquaculture, and global climate change. Improvements in toxin detection methods and increased toxin surveillance programmes are positive developments in limiting human exposure to shellfish toxins. PMID:20412612

  18. Functional RelBE-Family Toxin-Antitoxin Pairs Affect Biofilm Maturation and Intestine Colonization in Vibrio cholerae

    PubMed Central

    Hay, Amanda J.; Zhong, Zengtao; Zhu, Jun; Kan, Biao

    2015-01-01

    Toxin–antitoxin (TA) systems are small genetic elements that typically encode a stable toxin and its labile antitoxin. These cognate pairs are abundant in prokaryotes and have been shown to regulate various cellular functions. Vibrio cholerae, a human pathogen that is the causative agent of cholera, harbors at least thirteen TA loci. While functional HigBA, ParDE have been shown to stabilize plasmids and Phd/Doc to mediate cell death in V. cholerae, the function of seven RelBE-family TA systems is not understood. In this study we investigated the function of the RelBE TA systems in V. cholerae physiology and found that six of the seven relBE loci encoded functional toxins in E. coli. Deletion analyses of each relBE locus indicate that RelBE systems are involved in biofilm formation and reactive oxygen species (ROS) resistance. Interestingly, all seven relBE loci are induced under the standard virulence induction conditions and two of the relBE mutants displayed a colonization defect, which was not due to an effect on virulence gene expression. Although further studies are needed to characterize the mechanism of action, our study reveals that RelBE systems are important for V. cholerae physiology. PMID:26275048

  19. Strategies for the induction of immune responses at mucosal surfaces making use of cholera toxin B subunit as immunogen, carrier, and adjuvant.

    PubMed

    Holmgren, J; Czerkinsky, C; Lycke, N; Svennerholm, A M

    1994-01-01

    The concept of a common mucosal immune system, through which specific antigen-activated lymphocytes from the gut can disseminate immunity both along the intestinal tract and to various other mucosal and glandular tissues, has generated much current interest in the possibility of developing oral vaccines, not only for enteric infections but also for infections in the respiratory and urogenital tracts. However, to date it has proven difficult in practice to stimulate strong mucosal IgA immune responses by either parenteral or oral-mucosal administration of most antigens, and experience with soluble protein antigens has, on the whole, been disappointing. A notable exception in this regard is cholera toxin (CT) and in humans more than in other species, its nontoxic B subunit pentamer moiety (CTB). Based on this, CTB has become an important component in recently developed oral vaccines against cholera as well as against diarrhea caused by enterotoxigenic Escherichia coli producing CT-like heat-labile enterotoxin(s). Since the strong immunogenicity of CT and CTB can, to a large extent, be explained by their ability to bind to receptors on the intestinal mucosal surface, there has recently been much interest in approaches using CTB as an oral delivery carrier system for other vaccine-relevant antigens, and much progress has been made in preparing immunogenic hybrid proteins by coupling various protein or peptide antigens chemically or genetically to CTB. Indeed, in several systems, oral administration of such hybrid antigens has been found to markedly potentiate both intestinal and extraintestinal IgA immune responses against the CTB-coupled antigens and also to elicit substantial circulating antibody responses. Besides the mucosal immunopotentiating effect of either CT or CTB owing to their similar capacity as oral antigen-delivery vehicles, CT, but in most systems tested not CTB, also has strong adjuvant properties for stimulating mucosal IgA immune responses to admixed (not coupled) unrelated antigens after oral immunization. This adjuvant activity appears to be closely linked to the ADP-ribosylating action of CT (and specifically of its A subunit) leading to enhanced cyclic AMP formation in the affected cell, and efforts to eliminate the enterotoxic activity without losing adjuvanticity have so far not met with success. PMID:8203723

  20. EFFECTS OF MARINE ALGAL TOXINS ON THERMOREGULATION IN MICE.

    EPA Science Inventory

    Hypothermia is often seen in mice and rats exposed acutely to marine algal toxins, but the mechanism of action of these toxins on thermoregulation is not well understood. Our laboratory has assessed the thermoregulatory mechanisms of two marine algal toxins, maitotoxin and brevet...

  1. Pore-forming activity of clostridial binary toxins.

    PubMed

    Knapp, O; Benz, R; Popoff, M R

    2016-03-01

    Clostridial binary toxins (Clostridium perfringens Iota toxin, Clostridium difficile transferase, Clostridium spiroforme toxin, Clostridium botulinum C2 toxin) as Bacillus binary toxins, including Bacillus anthracis toxins consist of two independent proteins, one being the binding component which mediates the internalization into cell of the intracellularly active component. Clostridial binary toxins induce actin cytoskeleton disorganization through mono-ADP-ribosylation of globular actin and are responsible for enteric diseases. Clostridial and Bacillus binary toxins share structurally and functionally related binding components which recognize specific cell receptors, oligomerize, form pores in endocytic vesicle membrane, and mediate the transport of the enzymatic component into the cytosol. Binding components retain the global structure of pore-forming toxins (PFTs) from the cholesterol-dependent cytotoxin family such as perfringolysin. However, their pore-forming activity notably that of clostridial binding components is more related to that of heptameric PFT family including aerolysin and C. perfringens epsilon toxin. This review focuses upon pore-forming activity of clostridial binary toxins compared to other related PFTs. This article is part of a Special Issue entitled: Pore-Forming Toxins edited by Mauro Dalla Serra and Franco Gambale. PMID:26278641

  2. High-throughput production of two disulphide-bridge toxins.

    PubMed

    Upert, Grégory; Mourier, Gilles; Pastor, Alexandra; Verdenaud, Marion; Alili, Doria; Servent, Denis; Gilles, Nicolas

    2014-08-01

    A quick and efficient production method compatible with high-throughput screening was developed using 36 toxins belonging to four different families of two disulphide-bridge toxins. Final toxins were characterized using HPLC co-elution, CD and pharmacological studies. PMID:24947561

  3. Anthrax toxin-induced rupture of artificial lipid bilayer membranes

    NASA Astrophysics Data System (ADS)

    Nablo, Brian J.; Panchal, Rekha G.; Bavari, Sina; Nguyen, Tam L.; Gussio, Rick; Ribot, Wil; Friedlander, Art; Chabot, Donald; Reiner, Joseph E.; Robertson, Joseph W. F.; Balijepalli, Arvind; Halverson, Kelly M.; Kasianowicz, John J.

    2013-08-01

    We demonstrate experimentally that anthrax toxin complexes rupture artificial lipid bilayer membranes when isolated from the blood of infected animals. When the solution pH is temporally acidified to mimic that process in endosomes, recombinant anthrax toxin forms an irreversibly bound complex, which also destabilizes membranes. The results suggest an alternative mechanism for the translocation of anthrax toxin into the cytoplasm.

  4. Clostridium difficile toxin B is more potent than toxin A in damaging human colonic epithelium in vitro.

    PubMed Central

    Riegler, M; Sedivy, R; Pothoulakis, C; Hamilton, G; Zacherl, J; Bischof, G; Cosentini, E; Feil, W; Schiessel, R; LaMont, J T

    1995-01-01

    Toxin A but not toxin B, appears to mediate intestinal damage in animal models of Clostridium difficile enteritis. The purpose of this study was to investigate the electrophysiologic and morphologic effects of purified C. difficile toxins A and B on human colonic mucosa in Ussing chambers. Luminal exposure of tissues to 16-65 nM of toxin A and 0.2-29 nM of toxin B for 5 h caused dose-dependent epithelial damage. Potential difference, short-circuit current and resistance decreased by 76, 58, and 46%, respectively, with 32 nM of toxin A and by 76, 55, and 47%, respectively, with 3 nM of toxin B, when compared with baseline (P < 0.05). 3 nM of toxin A did not cause electrophysiologic changes. Permeability to [3H]mannitol increased 16-fold after exposure to 32 nM of toxin A and to 3 nM of toxin B when compared with controls (P < 0.05). Light and scanning electron microscopy after exposure to either toxin revealed patchy damage and exfoliation of superficial epithelial cells, while crypt epithelium remained intact. Fluorescent microscopy of phalloidin-stained sections showed that both toxins caused disruption and condensation of cellular F-actin. Our results demonstrate that the human colon is approximately 10 times more sensitive to the damaging effects of toxin B than toxin A, suggesting that toxin B may be more important than toxin A in the pathogenesis of C. difficile colitis in man. Images PMID:7738167

  5. Development and fabrication of heat-sterilizable inhalation therapy equipment

    NASA Technical Reports Server (NTRS)

    Irons, A. S.

    1974-01-01

    The development of a completely heat sterilizable intermittent positive pressure breathing (IPPB) ventilator in an effort to reduce the number of hospital acquired infections is reported. After appropriate changes in materials and design were made, six prototype units were fabricated and were successfully field tested in local hospitals. Most components of the modified ventilators are compatible with existing machines. In all but a few instances, such as installation of bacteria-retentive filters and a modified venturi, the change over from non-heat-sterilizable to sterilizable units was accomplished by replacement of heat labile materials with heat stable materials.

  6. Paralytic shellfish toxin profiles and toxin variability of the genus Alexandrium (Dinophyceae) isolated from the Southeast China Sea.

    PubMed

    Wang, Da-Zhi; Zhang, Shu-Gang; Gu, Hai-Feng; Chan, Leo Lai; Hong, Hua-Sheng

    2006-08-01

    Paralytic shellfish toxin (PST) profiles of 16 Alexandrium isolates from the Southeast China Sea were analyzed by high-pressure liquid chromatography. Toxin content and composition of three A. tamarense isolates, ATDH01, ATGX02 and ATMJ02, were also investigated at different growth phases and under various culture conditions. Our results showed that six strains of A. affine were non-toxic, while 10 strains of A. tamarense and A. catenella were toxic. These toxic isolates grown in the same culture conditions consistently produced an unusually high proportion of the N-sulfocarbamoyl toxin C1/2 (around 60-80% of total toxins) and medium amounts of gonyautoxin GTX5 (around 15-30% of total) with only trace quantities (<5% of total) of other saxitoxin derivatives (i.e. GTX1, GTX3, GTX4 and neoSTX). The toxin composition of three A. tamarense isolates did not vary with the growth phases, although higher toxin contents (Qt, fmolcell(-1)) were found in the exponential phase. Variations in temperature, salinity and nutrient levels affected toxin content of three A. tamarense isolates but they did not have pronounced effects on the toxin composition (mole %). These results indicate that toxin composition remained relatively constant under various culture conditions, suggesting that toxin composition could be used as a stable biomarker for the Alexandrium species in this region. However, comparison of toxin profiles between isolates from different localities require special caution since isolates even from the same region can have distinct toxin profiles. PMID:16859722

  7. A Cell-Based Fluorescent Assay to Detect the Activity of Shiga Toxin and Other Toxins That Inhibit Protein Synthesis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Escherichia coli O157:H7, a major cause of food-borne illness, produces Shiga toxins that block protein synthesis by inactivating the ribosome. In this chapter we describe a simple cell-based fluorescent assay to detect Shiga toxins and inhibitors of toxin activity. The assay can also be used to d...

  8. Okadaic Acid: More than a Diarrheic Toxin

    PubMed Central

    Valdiglesias, Vanessa; Prego-Faraldo, María Verónica; Pásaro, Eduardo; Méndez, Josefina; Laffon, Blanca

    2013-01-01

    Okadaic acid (OA) is one of the most frequent and worldwide distributed marine toxins. It is easily accumulated by shellfish, mainly bivalve mollusks and fish, and, subsequently, can be consumed by humans causing alimentary intoxications. OA is the main representative diarrheic shellfish poisoning (DSP) toxin and its ingestion induces gastrointestinal symptoms, although it is not considered lethal. At the molecular level, OA is a specific inhibitor of several types of serine/threonine protein phosphatases and a tumor promoter in animal carcinogenesis experiments. In the last few decades, the potential toxic effects of OA, beyond its role as a DSP toxin, have been investigated in a number of studies. Alterations in DNA and cellular components, as well as effects on immune and nervous system, and even on embryonic development, have been increasingly reported. In this manuscript, results from all these studies are compiled and reviewed to clarify the role of this toxin not only as a DSP inductor but also as cause of alterations at the cellular and molecular levels, and to highlight the relevance of biomonitoring its effects on human health. Despite further investigations are required to elucidate OA mechanisms of action, toxicokinetics, and harmful effects, there are enough evidences illustrating its toxicity, not related to DSP induction, and, consequently, supporting a revision of the current regulation on OA levels in food. PMID:24184795

  9. Innovative techniques for harmful algal toxin analysis.

    PubMed

    Pierce, R H; Kirkpatrick, G J

    2001-01-01

    The global increase in frequency and intensity of harmful algal blooms (HABs) has led to more frequent incidence of seafood-borne illnesses and adverse impacts on natural resources. In response, public health agencies worldwide have mobilized to initiate HAB monitoring programs. To meet this demand, innovative analytical techniques are being developed that provide rapid and reliable detection of the causative organisms and the toxins produced. Modifications to conventional chromatography and mass spectrometry have greatly improved sensitivity and selectivity of these methods toward naturally occurring phycotoxins. Bioassay techniques using live organisms are giving way to molecular and cellular methods that measure the toxicologically significant activity of the toxin molecules. Molecular probes are being applied to distinguish species-specific RNA and DNA sequences for rapid identification of HAB-causing organisms. The direction of this new technology is to develop rapid and reliable screening methods for phycotoxins and the causative organisms to provide protection for public health, aquaculture, and natural resources. New methods also are being developed for detecting minute amounts of toxin molecules in microenvironments, leading to understanding the toxicokinetics and toxicological functions of the toxins. PMID:11351396

  10. Toxins of molds from decaying tomato fruit.

    PubMed Central

    Harwig, J; Scott, P M; Stoltz, D R; Blanchfield, B J

    1979-01-01

    Among 27 mold isolates from decaying tomatoes, culture filtrates or ethyl acetate extracts of 8 isolates grown in yeast extract-sucrose medium were markedly toxic (mortality, greater than 50%) to brine shrimp larvae. The toxicity of six of these isolates could be attributed to the presence of citrinin, tenuazonic acid, or T-2 toxin. Ethyl acetate extracts of five Alternaria isolates and one Fusarium isolate were mutagenic for Salmonella typhimurium strains. In ripe tomatoes inoculated with toxin-producing isolates and incubated at 25 degrees C, one Alternaria alternata isolate produced tenuazonic acid in seven of seven tomatoes at levels of up to 106 micrograms/g and alternariol methyl ether in one of the seven tomatoes at 0.8 microgram/g. Another A. alternata isolate produced tenuazonic acid or alternariol methyl ether at much lower levels in only three of seven tomatoes. Patulin and citrinin were produced by a Penicillium expansum isolate at levels of up to 8.4 and 0.76 microgram/g, respectively. In tomatoes incubated at 15 degrees C, a Fusarium sulphureum isolate produced T-2 toxin, HT-2 toxin, and neosolaniol at levels of up to 37.5, 37.8 and 5.6 micrograms/g, respectively. If these mycotoxins are thermostable, they may occur at detectable levels in tomato products whenever partially moldy tomatoes are used as raw material. PMID:391152

  11. Botulinum toxin for treatment of Harlequin syndrome.

    PubMed

    Manhes, Roberta K J V; Spitz, Mariana; Vasconcellos, Luiz Felipe

    2016-02-01

    We described a patient with Harlequin syndrome, a rare neurological condition, characterized by unilateral facial sweating and flushing, who had a good response to botulinum toxin application. She had been submitted to sympathectomy a few years, however she still complained of excessive sweating in the regions mentioned and intense flushing. PMID:26750113

  12. Pasteurella multocida Toxin Manipulates T Cell Differentiation

    PubMed Central

    Hildebrand, Dagmar; Heeg, Klaus; Kubatzky, Katharina F.

    2015-01-01

    Pasteurella multocida causes various diseases in a broad range of wild and domestic animals. Toxigenic strains of the serotypes A and D produce an AB protein toxin named Pasteurella multocida toxin (PMT). PMT constitutively activates the heterotrimeric G protein subunits Gαq, Gα13, and Gαi through deamidation of a glutamine residue, which results in cytoskeletal rearrangements as well as increased proliferation and survival of the host cell. In human monocytes, PMT alters the lipopolysaccharide (LPS)-induced activation toward a phenotype that suppresses T cell activation. Here we describe that the toxin also modulates CD4-positive T helper (Th) cells directly. PMT amplifies the expansion of Th cells through enhanced cell cycle progression and suppression of apoptosis and manipulates the differentiation of Th subclasses through activation of Signal Transducers and Activators of Transcription (STAT) family members and induction of subtype-specific master transcription factors. A large population of toxin-treated T cells is double-positive for Foxp3 and RORγt, the transcription factors expressed by Treg and Th17 cells, respectively. This suggests that these cells could have the potential to turn into Th17 cells or suppressive Treg cells. However, in terms of function, the PMT-differentiated cells behave as inflammatory Th17 cells that produce IL-17 and trigger T cell proliferation. PMID:26635744

  13. (-)-Botryodiplodin, A Unique Ribose Analog Toxin

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Many toxins owe their mechanisms of action to being structural analogs of essential metabolites, messengers or structural components. Examples range from tubo-curare to penicillin. Ribose plays a unique role in the metabolism of living organisms, whether prokaryotes or eukaryotes. It and its deri...

  14. Cyanobacterial toxins: biosynthetic routes and evolutionary roots.

    PubMed

    Dittmann, Elke; Fewer, David P; Neilan, Brett A

    2013-01-01

    Cyanobacteria produce an unparalleled variety of toxins that can cause severe health problems or even death in humans, and wild or domestic animals. In the last decade, biosynthetic pathways have been assigned to the majority of the known toxin families. This review summarizes current knowledge about the enzymatic basis for the production of the hepatotoxins microcystin and nodularin, the cytotoxin cylindrospermopsin, the neurotoxins anatoxin and saxitoxin, and the dermatotoxin lyngbyatoxin. Elucidation of the biosynthetic pathways of the toxins has paved the way for the development of molecular techniques for the detection and quantification of the producing cyanobacteria in different environments. Phylogenetic analyses of related clusters from a large number of strains has also allowed for the reconstruction of the evolutionary scenarios that have led to the emergence, diversification, and loss of such gene clusters in different strains and genera of cyanobacteria. Advances in the understanding of toxin biosynthesis and evolution have provided new methods for drinking-water quality control and may inspire the development of techniques for the management of bloom formation in the future. PMID:23051004

  15. Future Avenues to Decrease Uremic Toxin Concentration.

    PubMed

    Vanholder, Raymond C; Eloot, Sunny; Glorieux, Griet L R L

    2016-04-01

    In this article, we review approaches for decreasing uremic solute concentrations in chronic kidney disease and in particular, in end-stage renal disease (ESRD). The rationale to do so is the straightforward relation between concentration and biological (toxic) effect for most toxins. The first section is devoted to extracorporeal strategies (kidney replacement therapy). In the context of high-flux hemodialysis and hemodiafiltration, we discuss increasing dialyzer blood and dialysate flows, frequent and/or extended dialysis, adsorption, bioartificial kidney, and changing physical conditions within the dialyzer (especially for protein-bound toxins). The next section focuses on the intestinal generation of uremic toxins, which in return is stimulated by uremic conditions. Therapeutic options are probiotics, prebiotics, synbiotics, and intestinal sorbents. Current data are conflicting, and these issues need further study before useful therapeutic concepts are developed. The following section is devoted to preservation of (residual) kidney function. Although many therapeutic options may overlap with therapies provided before ESRD, we focus on specific aspects of ESRD treatment, such as the risks of too-strict blood pressure and glycemic regulation and hemodynamic changes during dialysis. Finally, some recommendations are given on how research might be organized with regard to uremic toxins and their effects, removal, and impact on outcomes of uremic patients. PMID:26500179

  16. Toxins and antimicrobial peptides: Interactions with membranes

    PubMed Central

    Schlamadinger, Diana E.; Gable, Jonathan E.; Kim, Judy E.

    2014-01-01

    The innate immunity to pathogenic invasion of organisms in the plant and animal kingdoms relies upon cationic antimicrobial peptides (AMPs) as the first line of defense. In addition to these natural peptide antibiotics, similar cationic peptides, such as the bee venom toxin melittin, act as nonspecific toxins. Molecular details of AMP and peptide toxin action are not known, but the universal function of these peptides to disrupt cell membranes of pathogenic bacteria (AMPs) or a diverse set of eukaryotes and prokaryotes (melittin) is widely accepted. Here, we have utilized spectroscopic techniques to elucidate peptide-membrane interactions of alpha-helical human and mouse AMPs of the cathelicidin family as well as the peptide toxin melittin. The activity of these natural peptides and their engineered analogs was studied on eukaryotic and prokaryotic membrane mimics consisting of <200-nm bilayer vesicles composed of anionic and neutral lipids as well as cholesterol. Vesicle disruption, or peptide potency, was monitored with a sensitive fluorescence leakage assay. Detailed molecular information on peptide-membrane interactions and peptide structure was further gained through vibrational spectroscopy combined with circular dichroism. Finally, steady-state fluorescence experiments yielded insight into the local environment of native or engineered tryptophan residues in melittin and human cathelicidin embedded in bilayer vesicles. Collectively, our results provide clues to the functional structures of the engineered and toxic peptides and may impact the design of synthetic antibiotic peptides that can be used against the growing number of antibiotic-resistant pathogens. PMID:25593677

  17. Toxins and antimicrobial peptides: interactions with membranes

    NASA Astrophysics Data System (ADS)

    Schlamadinger, Diana E.; Gable, Jonathan E.; Kim, Judy E.

    2009-08-01

    The innate immunity to pathogenic invasion of organisms in the plant and animal kingdoms relies upon cationic antimicrobial peptides (AMPs) as the first line of defense. In addition to these natural peptide antibiotics, similar cationic peptides, such as the bee venom toxin melittin, act as nonspecific toxins. Molecular details of AMP and peptide toxin action are not known, but the universal function of these peptides to disrupt cell membranes of pathogenic bacteria (AMPs) or a diverse set of eukaryotes and prokaryotes (melittin) is widely accepted. Here, we have utilized spectroscopic techniques to elucidate peptide-membrane interactions of alpha-helical human and mouse AMPs of the cathelicidin family as well as the peptide toxin melittin. The activity of these natural peptides and their engineered analogs was studied on eukaryotic and prokaryotic membrane mimics consisting of <200-nm bilayer vesicles composed of anionic and neutral lipids as well as cholesterol. Vesicle disruption, or peptide potency, was monitored with a sensitive fluorescence leakage assay. Detailed molecular information on peptidemembrane interactions and peptide structure was further gained through vibrational spectroscopy combined with circular dichroism. Finally, steady-state fluorescence experiments yielded insight into the local environment of native or engineered tryptophan residues in melittin and human cathelicidin embedded in bilayer vesicles. Collectively, our results provide clues to the functional structures of the engineered and toxic peptides and may impact the design of synthetic antibiotic peptides that can be used against the growing number of antibiotic-resistant pathogens.

  18. Host specific toxins; effectors of necrotrophic pathogenicity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Host-specific toxins are defined as pathogen effectors that induce toxicity and promote disease only in the host species and only in cultivars of that host, and with few exceptions, expressing a specific dominant susceptibility gene. They are a feature of a small but well studied group of fungal pl...

  19. [From biotechnology to bacterial toxins and immunomodulators].

    PubMed

    Bizzini, B

    1984-03-01

    The disposal of industrial wastes has been recognized as a serious problem long before the ecological movement gained its present strength. The possibility of using "useful" bacteria for this purpose was considered rather early and numerous studies were made to use them for the biological treatment of industrial wastes. Such attempts have taken advantage of the development of large scale fermentors for the production of bacteria while at the same time they have stimulated progress in the technology of fermentors. The advances made in the technology of fermentors have also made possible the large scale production of pathogenic bacteria. This has made possible the production of bacterial toxins in adequate amounts, enabling investigators to purify them to homogeneity, to characterize the purified toxins physico-chemically and immunologically and to study their structure-function relationships. Similarly, large quantities of bacteria exhibiting specific biological activities could be produced and the constituents responsible for the activities of these bacteria could be isolated and characterized. In this paper, the Author describes the stages of his progress and processes through the fields of the biotechnology and toxinology to that of immunomodulators. His investigations have largely taken advantage of developments in the technology of fermentors. "Ecologic" bacteria have been isolated which are capable of using phenol as the only source of carbon or cyanides as the sole source of carbon and nitrogen. These bacteria have been used for the treatment of industrial wastes enriched either with phenol or with cyanides. Another type of "useful" bacteria investigated was Bifidibacterium bifidus which has a fundamental role in the economy of the breast-fed infant. B. bifidus is present only in the gut of infants fed with mothers-milk. Growth factors for B. bifidus have been isolated from casein hydrolyzates which permitted the implantation of B. bifidus in the gut of infants fed with cows-milk supplemented with these factors. A further development of this type of research has been the large scale production of toxigenic bacteria. As a result large quantities of tetanus toxin have been produced and a method of purification has been devised. The availability of large amounts of purified tetanus toxin made it possible to determine the physico-chemical characteristics of the toxin as well as some of its structure-function relationships. In addition, the mechanism of detoxification of protein toxins by the action of formaldehyde has been clarified. Enzymatically derived toxin fragments have also been purified.(ABSTRACT TRUNCATED AT 400 WORDS) PMID:6383425

  20. Higher cytotoxicity of divalent antibody-toxins than monovalent antibody-toxins

    SciTech Connect

    Won, JaeSeon; Nam, PilWon; Lee, YongChan; Choe, MuHyeon

    2009-04-24

    Recombinant antibody-toxins are constructed via the fusion of a 'carcinoma-specific' antibody fragment to a toxin. Due to the high affinity and high selectivity of the antibody fragments, antibody-toxins can bind to surface antigens on cancer cells and kill them without harming normal cells [L.H. Pai, J.K. Batra, D.J. FitzGerald, M.C. Willingham, I. Pastan, Anti-tumor activities of immunotoxins made of monoclonal antibody B3 and various forms of Pseudomonas exotoxin, Proc. Natl. Acad. Sci. USA 88 (1991) 3358-3362]. In this study, we constructed the antibody-toxin, Fab-SWn-PE38, with SWn (n = 3, 6, 9) sequences containing n-time repeated (G{sub 4}S) between the Fab fragment and PE38 (38 kDa truncated form of Pseudomonas exotoxin A). The SWn sequence also harbored one cysteine residue that could form a disulfide bridge between two Fab-SWn-PE38 monomers. We assessed the cytotoxicity of the monovalent (Fab-SWn-PE38), and divalent ([Fab-SWn-PE38]{sub 2}) antibody-toxins. The cytotoxicity of the dimer against the CRL1739 cell line was approximately 18.8-fold higher than that of the monomer on the ng/ml scale, which was approximately 37.6-fold higher on the pM scale. These results strongly indicate that divalency provides higher cytotoxicity for an antibody-toxin.

  1. Neutralizing Monoclonal Antibodies against Disparate Epitopes on Ricin Toxin's Enzymatic Subunit Interfere with Intracellular Toxin Transport.

    PubMed

    Yermakova, Anastasiya; Klokk, Tove Irene; O'Hara, Joanne M; Cole, Richard; Sandvig, Kirsten; Mantis, Nicholas J

    2016-01-01

    Ricin is a member of the A-B family of bacterial and plant toxins that exploit retrograde trafficking to the Golgi apparatus and endoplasmic reticulum (ER) as a means to deliver their cytotoxic enzymatic subunits into the cytoplasm of mammalian cells. In this study we demonstrate that R70 and SyH7, two well-characterized monoclonal antibodies (mAbs) directed against distinct epitopes on the surface of ricin's enzymatic subunit (RTA), interfere with toxin transport from the plasma membrane to the trans Golgi network. Toxin-mAb complexes formed on the cell surface delayed ricin's egress from EEA-1(+) and Rab7(+) vesicles and enhanced toxin accumulation in LAMP-1(+) vesicles, suggesting the complexes were destined for degradation in lysosomes. Three other RTA-specific neutralizing mAbs against different epitopes were similar to R70 and SyH7 in terms of their effects on ricin retrograde transport. We conclude that interference with toxin retrograde transport may be a hallmark of toxin-neutralizing antibodies directed against disparate epitopes on RTA. PMID:26949061

  2. Toxin Instability and Its Role in Toxin Translocation from the Endoplasmic Reticulum to the Cytosol

    PubMed Central

    Teter, Ken

    2013-01-01

    AB toxins enter a host cell by receptor-mediated endocytosis. The catalytic A chain then crosses the endosome or endoplasmic reticulum (ER) membrane to reach its cytosolic target. Dissociation of the A chain from the cell-binding B chain occurs before or during translocation to the cytosol, and only the A chain enters the cytosol. In some cases, AB subunit dissociation is facilitated by the unique physiology and function of the ER. The A chains of these ER-translocating toxins are stable within the architecture of the AB holotoxin, but toxin disassembly results in spontaneous or assisted unfolding of the isolated A chain. This unfolding event places the A chain in a translocation-competent conformation that promotes its export to the cytosol through the quality control mechanism of ER-associated degradation. A lack of lysine residues for ubiquitin conjugation protects the exported A chain from degradation by the ubiquitin-proteasome system, and an interaction with host factors allows the cytosolic toxin to regain a folded, active state. The intrinsic instability of the toxin A chain thus influences multiple steps of the intoxication process. This review will focus on the hosttoxin interactions involved with A chain unfolding in the ER and A chain refolding in the cytosol. PMID:24970201

  3. Mutant with diphtheria toxin receptor and acidification function but defective in entry of toxin

    SciTech Connect

    Kohno, Kenji ); Hayes, H.; Mekada, Eisuke ); Uchida, Tsuyoshi Osaka Univ. )

    1987-09-01

    A mutant of Chinese hamster ovary cells, GE1, that is highly resistant to diphtheria toxin was isolated. The mutant contains 50% ADP-ribosylatable elongation factor 2, but its protein synthesis was not inhibited by the toxin even at concentrations above 100 {mu}g/ml. {sup 125}I-labeled diphtheria toxin was associated with GE1 cells as well as with the parent cells but did not block protein synthesis of GE1 cells even when the cells were exposed to low pH in the presence or absence of NH{sub 4}Cl. The infections of GE1 cells and the parent cells by vesicular stomatitis virus were similar. GE1 cells were cross-resistant to Pseudomonas aeruginosa exotoxin A and so were about 1,000 times more resistant to this toxin than the parent cells. Hybrids of GE1 cells and the parent cells or mutant cells lacking a functional receptor were more sensitive to diphtheria toxin than GE1 cells. These results suggest that entry of diphtheria toxin into cells requires a cellular factor(s) in addition to those involved in receptor function and acidification of endosomes and that GE1 cells do not express this cellular factor. This character is recessive in GE1 cells.

  4. Functional and Structural Insights of a Staphylococcus aureus Apoptotic-like Membrane Peptide from a Toxin-Antitoxin Module*

    PubMed Central

    Sayed, Nour; Nonin-Lecomte, Sylvie; Réty, Stéphane; Felden, Brice

    2012-01-01

    We report a functional type I toxin-antitoxin (TA) module expressed by a human pathogen, Staphylococcus aureus. TA systems consist of stable toxins and labile antitoxins encoded within small genetic modules widespread in eubacteria and archaea. TA genes provide stress adaptation and protection against DNA loss or invasion. The genes encoding the SprA1 toxic peptide (PepA1) and the SprA1AS RNA antitoxin are within a pathogenicity island on opposite strands and possess a 3′ overlap. To prevent peptide toxicity during S. aureus growth, PepA1 expression from stable (half-life > 3 h) SprA1 is repressed by elevated amounts of unstable (half-life = ∼10 mn) SprA1AS. In vivo, PepA1 localizes at the bacterial membrane and triggers S. aureus death. Based on NMR and CD data, its solution structure was solved and is a long bent, interrupted helix. Molecular dynamics simulations indicate that PepA1 compaction and helical content fluctuate in accordance with its cytoplasm or membrane location. When inserted into the S. aureus membrane, the PepA1 conformation switches to a ∼7-nm-long continuous helix, presumably forming pores to alter membrane integrity. PepA1 expression is induced upon acidic and oxidative stresses by reducing SprA1AS levels. As an altruistic behavior during infection, some cells may induce the expression of that toxin that would facilitate departure from the host immune cells for spreading. PMID:23129767

  5. Glycolipids as receptors for Bacillus thuringiensis crystal toxin.

    PubMed

    Griffitts, Joel S; Haslam, Stuart M; Yang, Tinglu; Garczynski, Stephan F; Mulloy, Barbara; Morris, Howard; Cremer, Paul S; Dell, Anne; Adang, Michael J; Aroian, Raffi V

    2005-02-11

    The development of pest resistance threatens the effectiveness of Bacillus thuringiensis (Bt) toxins used in transgenic and organic farming. Here, we demonstrate that (i) the major mechanism for Bt toxin resistance in Caenorhabditis elegans entails a loss of glycolipid carbohydrates; (ii) Bt toxin directly and specifically binds glycolipids; and (iii) this binding is carbohydrate-dependent and relevant for toxin action in vivo. These carbohydrates contain the arthroseries core conserved in insects and nematodes but lacking in vertebrates. We present evidence that insect glycolipids are also receptors for Bt toxin. PMID:15705852

  6. Preservation of labile organic matter in soils of drained thaw lakes in Northern Alaska

    NASA Astrophysics Data System (ADS)

    Mueller, Carsten W.; Rethemeyer, Janet; Kao-Kniffin, Jenny; Lppmann, Sebastian; Hinkel, Kenneth; Bockheim, James

    2014-05-01

    A large number of studies predict changing organic matter (OM) dynamics in arctic soils due to global warming. In contrast to rather slowly altering bulk soil properties, single soil organic matter (SOM) fractions can provide a more detailed picture of the dynamics of differently preserved SOM pools in climate sensitive arctic regions. By the study of the chemical composition of such distinctive SOM fractions using nuclear magnetic resonance spectroscopy (NMR) together with radiocarbon analyses it is possible to evaluate the stability of the major OM pools. Approximately 50-75% of Alaska's Arctic Coastal Plain is covered with thaw lakes and drained thaw lakes that follow a 5,000 yr cycle of development (between creation and final drainage), thus forming a natural soil chronosequence. The drained thaw lakes offer the possibility to study SOM dynamics affected by permafrost processes over millennial timescales. In April 2010 we sampled 16 soil cores (including the active and permanent layer) reaching from young drained lakes (0-50 years since drainage) to ancient drained lakes (3000-5500 years since drainage). Air dried soil samples from soil horizons of the active and permanent layer were subjected to density fractionation in order to differentiate particulate OM and mineral associated OM. The chemical composition of the SOM fractions was analyzed by 13C CPMAS NMR spectroscopy. For a soil core of a young and an ancient drained thaw lake basin we also analyzed the 14C content. For the studied soils we can show that up to over 25 kg OC per square meter are stored mostly as labile, easily degradable organic matter rich in carbohydrates. In contrast only 10 kg OC per square meter were sequestered as presumably more stable mineral associated OC dominated by aliphatic compounds. Comparable to soils of temperate regions, we found small POM (< 20 m) occluded in aggregated soil structures which differed in the chemical composition from larger organic particles. This was clearly shown by increased amounts of aliphatic C in these small POM fractions. As revealed by 13C CPMAS NMR, with advancing soil age increasing aliphaticity was also detected in occluded small POM fractions. By 14C dating we could show the stabilization of younger more labile OM at greater depth in buried O horizons. Additionally the study of the microscale elemental distributions, using nano-scale secondary ion mass spectrometry (NanoSIMS) showed the initial formation of aggregates and organo-mineral interfaces in the studied permafrost soils.

  7. Ultrafast photochemistry of polyatomic molecules containing labile halogen atoms in solution

    NASA Astrophysics Data System (ADS)

    Mereshchenko, Andrey S.

    Because breaking and making of chemical bonds lies at the heart of chemistry, this thesis focuses on dynamic studies of labile molecules in solutions using ultrafast transient absorption spectroscopy. Specifically, my interest is two-fold: (i) novel reaction intermediates of polyhalogenated carbon, boron and phosphorus compounds; (ii) photophysics and photochemistry of labile copper(II) halide complexes. Excitation of CH2Br2, CHBr3, BBr 3, and PBr3 into n(Br)sigma*(X-Br) states, where X=C, B, or P, leads to direct photoisomerization with formation of isomers having Br-Br bonds as well as rupture of one of X-Br bonds with the formation of a Br atom and a polyatomic radical fragment, which subsequently recombine to form similar isomer products. Nonpolar solvation stabilizes the isomers, consistent with intrinsic reaction coordinate calculations of the isomer ground state potential energy surfaces at the density functional level of theory, and consequently, the involvement of these highly energetic species on chemically-relevant time scales needs to be taken into account. Monochlorocomplexes in methanol solutions promoted to the ligand-to-metal charge transfer (LMCT) excited state predominantly undergo internal conversion via back electron transfer, giving rise to vibrationally hot ground-state parent complexes. Copper-chloride homolitical bond dissociation yielding the solvated copper(I) and Cl- atom/solvent CT complexes constitutes a minor pathway. Insights into ligand substitution mechanisms were acquired by monitoring the recovery of monochloro complexes at the expense of two unexcited dichloro- and unsubstituted forms of Cu(II) complexes also present in the solution. Detailed description of ultrafast excited-state dynamics of CuCl 42- complexes in acetonitrile upon excitation into all possible Ligand Field (LF) excited states and two most intense LMCT transitions is reported. The LF states were found to be nonreactive with lifetimes remarkably longer than those for copper(II) complexes studied so far, in particular, copper blue proteins. The highest 2A1 and lowest 2E LF states relax directly to the ground electronic state whereas the intermediate 2B1 LF state relaxes stepwise through the 2E state. The LMCT excited states are short-lived undergoing either ionic dissociation (CuCl3- + Cl-) or cascading relaxation through the manifold of vibrationally hot LF states to the ground state.

  8. Acid-labile sulfides in shallow marine bottom sediments: A review of the impact on ecosystems in the Azov Sea, the NE Black Sea shelf and NW Adriatic lagoons

    NASA Astrophysics Data System (ADS)

    Sorokin, Yu. I.; Zakuskina, O. Yu

    2012-02-01

    Acid-labile sulfides (LS) increase in bottom sediments at sites in the Azov Sea, at the NE Black Sea shelf and in the coastal lagoons of NW Adriatic Sea experiencing direct impacts of anthropogenic pollution. Fresh anthropogenic organic matter stimulates the bacterial sulfate reduction and here the rate of the LS production overcomes their loss during the oxidation and pyritization. This results in the expansion of reduced sediment layer up to the bottom surface. The LS concentration in the reduced sediments varies between 300 and 2000 mg S l -1 of wet silt depending on the size of pollution loading and on the rate of sedimentation. In the oxidized sediments away from the direct pollution impact, the LS concentration did not exceed 100-150 mg S l -1. Being a strong cytochrome toxin, the LS adversely affect the coastal ecosystems. The concentrations over 600 mg S l -1 result in quasi total benthic mortality whereas >300-400 mg S l -1 depletes the benthic faunal abundance and taxonomic diversity. Accumulation of the LS in sediments also induces nocturnal hypoxia and stimulates domination of toxic cyanobacteria in the pelagic phytocenoses.

  9. Evaluating Multiple Drivers of Soil Organic Matter Lability and Structure in a Sub-Alpine Forest Ecosystem

    NASA Astrophysics Data System (ADS)

    Hall, E.; Fegel, T. S., II; Boot, C. M.

    2014-12-01

    Aeolian deposition of reactive nitrogen (N) is reaching even the most remote ecosystems. There has been an abundance of research investigating how these subsidies of reactive N may alter fundamental ecosystem characteristics such as soil organic matter (SOM) pool size. Previous studies have reported that additions of reactive N have the potential to both increase and decrease SOM content. While there are a series of different variables that may affect the size of the SOM pool it has been suggested that the lability or recalcitrance of the SOM may be related to its chemical composition (kind and relative abundance of constituent molecules). To address this we sampled 6 experimental plots in a sub-alpine forest in Rocky Mountain National Park (3 control and 3 treated with reactive N for 18 years) during two months in the summers of 2011 and 2012. We found the SOM content of the control plots was greater than that of the experimental plots. To assess lability of each SOM sample we extracted the SOM from each plot with water and incubated the dissolved organic carbon with a common aquatic microbial community from a lake within the watershed. To assess structure of the SOM pool we used ultra performance liquid chromatography (UPLC) coupled with MS of each extract before incubation with the bacterial community. The dissolved component of the SOM showed clear differences in lability both in total quantity and rate of decomposition during incubation with aquatic microorganisms. Principle components analysis indicated season was a stronger driver of DOM composition than fertilization, describing the majority of the variability between July and September 2012. When samples were considered within a season and year there were additional differences in both lability and composition of DOM. Here we evaluate the relative influence of inter- and intra-annual variability and reactive N on both the characteristics and composition of SOM. By linking UPLC-MS with a functional assay of lability we attempt to define chemical characteristics of lability that can be assessed across ecosystems. Doing so will allow us to better understand linked biogeochemical cycles (C and N) across a wide range of soil ecosystems.

  10. Botulinum Toxin: Pharmacology and Therapeutic Roles in Pain States.

    PubMed

    Patil, Shilpadevi; Willett, Olga; Thompkins, Terin; Hermann, Robert; Ramanathan, Sathish; Cornett, Elyse M; Fox, Charles J; Kaye, Alan David

    2016-03-01

    Botulinum toxin, also known as Botox, is produced by Clostridium botulinum, a gram-positive anaerobic bacterium, and botulinum toxin injections are among the most commonly practiced cosmetic procedures in the USA. Although botulinum toxin is typically associated with cosmetic procedures, it can be used to treat a variety of other conditions, including pain. Botulinum toxin blocks the release of acetylcholine from nerve endings to paralyze muscles and to decrease the pain response. Botulinum toxin has a long duration of action, lasting up to 5 months after initial treatment which makes it an excellent treatment for chronic pain patients. This manuscript will outline in detail why botulinum toxin is used as a successful treatment for pain in multiple conditions as well as outline the risks associated with using botulinum toxin in certain individuals. As of today, the only FDA-approved chronic condition that botulinum toxin can be used to treat is migraines and this is related to its ability to decrease muscle tension and increase muscle relaxation. Contraindications to botulinum toxin treatments are limited to a hypersensitivity to the toxin or an infection at the site of injection, and there are no known drug interactions with botulinum toxin. Botulinum toxin is an advantageous and effective alternative pain treatment and a therapy to consider for those that do not respond to opioid treatment. In summary, botulinum toxin is a relatively safe and effective treatment for individuals with certain pain conditions, including migraines. More research is warranted to elucidate chronic and long-term implications of botulinum toxin treatment as well as effects in pregnant, elderly, and adolescent patients. PMID:26879873

  11. Controls on the composition and lability of dissolved organic matter in Siberia's Kolyma River basin

    NASA Astrophysics Data System (ADS)

    Mann, P. J.; Davydova, A.; Zimov, N.; Spencer, R. G. M.; Davydov, S.; Bulygina, E.; Zimov, S.; Holmes, R. M.

    2012-03-01

    High-latitude northern rivers export globally significant quantities of dissolved organic carbon (DOC) to the Arctic Ocean. Climate change, and its associated impacts on hydrology and potential mobilization of ancient organic matter from permafrost, is likely to modify the flux, composition, and thus biogeochemical cycling and fate of exported DOC in the Arctic. This study examined DOC concentration and the composition of dissolved organic matter (DOM) across the hydrograph in Siberia's Kolyma River, with a particular focus on the spring freshet period when the majority of the annual DOC load is exported. The composition of DOM within the Kolyma basin was characterized using absorbance-derived measurements (absorbance coefficienta330, specific UV absorbance (SUVA254), and spectral slope ratio SR) and fluorescence spectroscopy (fluorescence index and excitation-emission matrices (EEMs)), including parallel factor analyses of EEMs. Increased surface runoff during the spring freshet led to DOM optical properties indicative of terrestrial soil inputs with high humic-like fluorescence, SUVA254, and low SRand fluorescence index (FI). Under-ice waters, in contrast, displayed opposing trends in optical properties representing less aromatic, lower molecular weight DOM. We demonstrate that substantial losses of DOC can occur via biological (30% over 28 days) and photochemical pathways (>29% over 14 days), particularly in samples collected during the spring freshet. The emerging view is therefore that of a more dynamic and labile carbon pool than previously thought, where DOM composition plays a fundamental role in controlling the fate and removal of DOC at a pan-Arctic scale.

  12. DGT measurement in low flow conditions: diffusive boundary layer and lability considerations.

    PubMed

    Uher, Emmanuelle; Tusseau-Vuillemin, Marie-Hlne; Gourlay-France, Catherine

    2013-07-01

    Recent papers have alerted the scientific community that a diffusive boundary layer (DBL) forming in front of diffusive gradients in thin film (DGT) devices when they are immersed in water might have a significant impact on the results and have suggested a method to assess the DBL. This paper aims at evaluating to what extent the DBL impacts the results of metal measurement in water by DGT and providing new information on the dissociation kinetics of metal complexes in wastewater by using DBL calculation. A careful study of the influence of the water velocity on the measurement with DGTs equipped with restricted gels is presented. Deployments took place in the laboratory with a range of stirring speeds (0-400 rpm) and in a canal receiving treated wastewater with increasing controlled water velocity (0.07-3 cm s(-1)). Even under extreme low flow conditions, the error made in using the equation that does not take into account that the DBL was lower than the analytical error. Nevertheless, the DBL is the seat of dissociation of complexes and increases the lability window beyond the steric constraints of the hydrogel. The capacity of restricted gels to only sample inorganic species under these conditions is questioned. This study also is an opportunity to provide information on metal-ligand interactions in wastewater by creating the kinetic signature of the wastewater. Unlike previous studies which used different types of water, Pb was the more limited metal and interacted strongly with the ligands. PMID:23722876

  13. Membrane Transport Behavior and the Lability of Chloride on Polyphosphazenes Bearing Bulky Substituents

    SciTech Connect

    Frederick F. Stewart; John R. Klaehn; Christopher J. Orme

    2007-08-01

    Polyphosphazenes are an intriguing class of inorganic polymers where much of their functionality is derived from pendant groups attached to phosphorus. The backbone of the polymer consists of alternating phosphorus and nitrogen atoms where the bonding is conventionally drawn as alternating double and single bonds. Orbital nodes are located at each phosphorus atom resulting in electron delocalization between phosphorus atoms, but not through them. Thus, the polymer backbone has a high degree of flexibility where halogens or other leaving groups can be effectively displaced with nucleophiles. In this paper, the first known example of a polyphosphazene with large quantities of non-labile chloride substituents induced by neighboring group steric effects will be discussed. This example is the result of the substitution of poly[bis-chlorophosphazene] with the sodium salt of 3,5-di-tert-butylphenol where only 60% of the chlorines were displaced. This contrasts with the 100% substitution observed with other phenols (phenol, 4-tert-butylphenol, 3-methylphenol, etc.).

  14. Sample storage-induced changes in the quantity and quality of soil labile organic carbon

    NASA Astrophysics Data System (ADS)

    Sun, Shou-Qin; Cai, Hui-Ying; Chang, Scott X.; Bhatti, Jagtar S.

    2015-11-01

    Effects of sample storage methods on the quantity and quality of labile soil organic carbon are not fully understood even though their effects on basic soil properties have been extensively studied. We studied the effects of air-drying and frozen storage on cold and hot water soluble organic carbon (WSOC). Cold- and hot-WSOC in air-dried and frozen-stored soils were linearly correlated with those in fresh soils, indicating that storage proportionally altered the extractability of soil organic carbon. Air-drying but not frozen storage increased the concentrations of cold-WSOC and carbohydrate in cold-WSOC, while both increased polyphenol concentrations. In contrast, only polyphenol concentration in hot-WSOC was increased by air-drying and frozen storage, suggesting that hot-WSOC was less affected by sample storage. The biodegradability of cold- but not hot-WSOC was increased by air-drying, while both air-drying and frozen storage increased humification index and changed specific UV absorbance of both cold- and hot-WSOC, indicating shifts in the quality of soil WSOC. Our results suggest that storage methods affect the quantity and quality of WSOC but not comparisons between samples, frozen storage is better than air-drying if samples have to be stored, and storage should be avoided whenever possible when studying the quantity and quality of both cold- and hot-WSOC.

  15. Sample storage-induced changes in the quantity and quality of soil labile organic carbon.

    PubMed

    Sun, Shou-Qin; Cai, Hui-Ying; Chang, Scott X; Bhatti, Jagtar S

    2015-01-01

    Effects of sample storage methods on the quantity and quality of labile soil organic carbon are not fully understood even though their effects on basic soil properties have been extensively studied. We studied the effects of air-drying and frozen storage on cold and hot water soluble organic carbon (WSOC). Cold- and hot-WSOC in air-dried and frozen-stored soils were linearly correlated with those in fresh soils, indicating that storage proportionally altered the extractability of soil organic carbon. Air-drying but not frozen storage increased the concentrations of cold-WSOC and carbohydrate in cold-WSOC, while both increased polyphenol concentrations. In contrast, only polyphenol concentration in hot-WSOC was increased by air-drying and frozen storage, suggesting that hot-WSOC was less affected by sample storage. The biodegradability of cold- but not hot-WSOC was increased by air-drying, while both air-drying and frozen storage increased humification index and changed specific UV absorbance of both cold- and hot-WSOC, indicating shifts in the quality of soil WSOC. Our results suggest that storage methods affect the quantity and quality of WSOC but not comparisons between samples, frozen storage is better than air-drying if samples have to be stored, and storage should be avoided whenever possible when studying the quantity and quality of both cold- and hot-WSOC. PMID:26617054

  16. Uncaria tomentosa (Willd. ex. Roem. & Schult.) DC. and Eucalyptus globulus Labill. interactions when administered with diazepam.

    PubMed

    Qulez, A M; Saenz, M T; Garca, M D

    2012-03-01

    The safety of natural drugs is defined by their side effects and toxicity as well as any interactions that may occur if taken together with other drugs. In particular, it is essential to identify synergies, antagonisms and other types of interference with other drugs so that the correct choice can be made from the range of phytomedicines available. The aim of this work was to investigate changes in the pharmacological effect of diazepam (2?mg/kg) on the CNS when administered together with a medicinal plant: Eucalyptus globulus Labill. (eucalyptus 6?mg/kg and 3.25?mg/kg) or Uncaria tomentosa (Willd. ex Roem. & Schult). DC. (cat's claw, 7.14?mg/kg and 3.54?mg/kg). Various different psychopharmacological effects were evaluated through assessing exploratory behavior, muscle relaxation and spontaneous motor activity. Both phytodrugs interacted with the benzodiazepine. Eucalyptus had an inhibitory effect at both doses and could be useful at the highest dose in cases where the desired effect of the depressant is moderate anxiolytic activity without marked muscle relaxation. Cat's claw, at both doses, enhanced the action of diazepam on spontaneous motor activity and, at the lowest dose, exploratory ability. These herbal drugs could be useful for their antiinflammatory activity in musculoskeletal pathologies treated with benzodiazepines. PMID:21928376

  17. [In Situ High-Resolution Analysis of Labile Phosphorus in Sediments of Lake Chaohu].

    PubMed

    Li, Chao; Wang, Dan; Yang, Jin-yan; Wang, Yan; Ding, Shi-ming

    2015-06-01

    High-resolution pore water equilibrators (HR-Peeper) and diffusive gradients in the thin films ( DGT) technologies were combined to in situ measure soluble reactive phosphorus (c(PW)) and labile phosphorus (C(DGT)) on seven sites in the west of Lake Chaohu. Vertical distributions of c(PW) and c(DGT) in most sediment profiles were similar to a different extent, demonstrating that the buffer capacity of the sediment solids to pore water SRP was similar at different depths. The diffusion flux across the sediment-water interface (SWI) and the ratio of cDGT/ c(PW) (R) were used to characterize phosphorus activity in sediments. From the center of the lake to the estuary of Nanfei River, the values of c(PW) and c(DGT) within the 6 mm layer were below the SWI and the SWI diffusion flux gradually increased, reflecting an increase in pollution level of sediment phosphorus. The change of R values was unconspicuous, indicating that the buffering capacity of the interface sediment had no significant difference. PMID:26387310

  18. Effect of pH on Metal Lability in Drinking Water Treatment Residuals.

    PubMed

    Wang, Changhui; Yuan, Nannan; Pei, Yuansheng

    2014-01-01

    Drinking water treatment residuals (WTRs), by-products generated during treatment of drinking water, can be reused as environmental amendments to remediate contamination. However, this beneficial reuse may be hampered by the potential release of toxic contaminants (e.g., metals) in the WTRs. In present study, batch tests and then fractionation, in vitro digestion, and the toxicity characteristic leaching procedure were used to investigate the release and extractability of metals in the Fe/Al hydroxides comprised WTRs under differing pH. The results demonstrated that significant release from WTRs for Ba, Be, Ca, Cd, Co, Cr, Fe, Mg, Mn, Pb, Sr, and Zn occurred under low pH (acid condition); for As, Mo, and V under high pH (alkaline condition); and for Al, Cu, and Ni under both conditions. In comparison, most metals in the WTRs were more easily released under low pH, but the release was stable at a relatively low level between pH 6 and 9, especially under alkaline conditions. Further analysis indicated that the chemical extractability and bioaccessibility of many metals was found to increase in the WTRs after being leached, even though the leached WTRs could still be considered nonhazardous. These results demonstrated that pH had a substantial effect on the lability of metals in WTRs. Overall, caution should be used when considering pH conditions during WTRs reuse to avoid potential metal pollution. PMID:25602573

  19. Sample storage-induced changes in the quantity and quality of soil labile organic carbon

    PubMed Central

    Sun, Shou-Qin; Cai, Hui-Ying; Chang, Scott X.; Bhatti, Jagtar S.

    2015-01-01

    Effects of sample storage methods on the quantity and quality of labile soil organic carbon are not fully understood even though their effects on basic soil properties have been extensively studied. We studied the effects of air-drying and frozen storage on cold and hot water soluble organic carbon (WSOC). Cold- and hot-WSOC in air-dried and frozen-stored soils were linearly correlated with those in fresh soils, indicating that storage proportionally altered the extractability of soil organic carbon. Air-drying but not frozen storage increased the concentrations of cold-WSOC and carbohydrate in cold-WSOC, while both increased polyphenol concentrations. In contrast, only polyphenol concentration in hot-WSOC was increased by air-drying and frozen storage, suggesting that hot-WSOC was less affected by sample storage. The biodegradability of cold- but not hot-WSOC was increased by air-drying, while both air-drying and frozen storage increased humification index and changed specific UV absorbance of both cold- and hot-WSOC, indicating shifts in the quality of soil WSOC. Our results suggest that storage methods affect the quantity and quality of WSOC but not comparisons between samples, frozen storage is better than air-drying if samples have to be stored, and storage should be avoided whenever possible when studying the quantity and quality of both cold- and hot-WSOC. PMID:26617054

  20. Labile Zn ions on octacalcium phosphate-derived Zn-containing hydroxyapatite surfaces

    NASA Astrophysics Data System (ADS)

    Honda, Yoshitomo; Anada, Takahisa; Morimoto, Shinji; Suzuki, Osamu

    2013-05-01

    We previously synthesized and characterized zinc-containing octacalcium phosphate (OCP) and its hydrolyzed Ca-deficient hydroxyapatite (HA). In the present report, we attempted to define the state of Zn in the OCP-derived Zn-calcium phosphates (CaPs) in relation to the presence of specific amino acids. Zn-containing OCPs were prepared in solutions that included Zn ions up to a concentration of 3.5 mM, and their hydrolyzates [hydrolyzed (hy)-Zn-CaP] were obtained in hot water. The materials were characterized by x-ray diffraction and scanning electron microscopy. The concentration of Ca and Zn ions at room temperature was determined by analyzing the supernatant after incubating the materials in ?-minimal essential medium (?-MEM) and HEPES buffer including cysteine, histidine, lysine, aspartic acid, and glutamic acid. Zn ions were more dissolved in ?-MEM than HEPES buffer in the absence of amino acids. The inclusion of the amino acids enhanced Zn dissolution by several hundred fold, even in HEPES buffer. Among the amino acids, both cysteine and histidine enhanced the release of Zn. The effect was particularly remarkable with cysteine even in the presence of the other amino acids tested. These results indicate that Zn ions are present as a surface labile pool, which tends to be preferentially desorbed by cysteine, a ubiquitous molecule present in serum.

  1. Stabilized liquid membrane device (SLMD) for the passive, integrative sampling of labile metals in water

    USGS Publications Warehouse

    Brumbaugh, W.G.; Petty, J.D.; Huckins, J.N.; Manahan, S.E.

    2002-01-01

    A stabilized liquid membrane device (SLMD) is described for potential use as an in situ, passive, integrative sampler for cadmium (Cd), cobalt (Co), copper (Cu), nickel (Ni), lead (Pb), and zinc (Zn) in natural waters. The SLMD (patent pending) consists of a 2.5-cm-wide by 15-cm-long strip of low-density polyethylene (LDPE) layflat tubing containing 1 mL of an equal mixture (v/v) of oleic acid (cis-9-octadecenoic acid) and EMO-8Q (7-[4-ethyl-1-methyloctyl]-8-quinolinol). The reagent mixture continuously diffuses to the exterior surface of the LDPE membrane, and provides for sequestration of several divalent metals for up to several weeks. Depending on sampler configuration, concentration factors of several thousand can be realized for these metal ions after just a few days. In addition to in situ deployment, the SLMD may be useful for laboratory determination of labile metal species in grab samples. Methods for minimizing the effects of water flow on the sampling rate are currently under investigation.

  2. Parasite maturation and host serum iron influence the labile iron pool of erythrocyte stage Plasmodium falciparum

    PubMed Central

    Clark, Martha; Fisher, Nancy C.; Kasthuri, Raj; Hand, Carla Cerami

    2014-01-01

    Summary Iron is a critical and tightly regulated nutrient for both the malaria parasite and its human host. The importance of the relationship between host iron and the parasite has been underscored recently by studies showing that host iron supplementation may increase the risk of falciparum malaria. It is unclear what host iron sources the parasite is able to access. We developed a flow cytometry-based method for measuring the labile iron pool (LIP) of parasitized erythrocytes using the nucleic acid dye STYO 61 and the iron sensitive dye, calcein acetoxymethyl ester (CA-AM). This new approach enabled us to measure the LIP of P. falciparum through the course of its erythrocytic life cycle and in response to the addition of host serum iron sources. We found that the LIP increases as the malaria parasite develops from early ring to late schizont stage, and that the addition of either transferrin or ferric citrate to culture media increases the LIP of trophozoites. Our method for detecting the LIP within malaria parasitized RBCs provides evidence that the parasite is able to access serum iron sources as part of the host vs. parasite arms race for iron. PMID:23398516

  3. Dissolved organic carbon lability and stable isotope shifts during microbial decomposition in a tropical river system

    NASA Astrophysics Data System (ADS)

    Geeraert, N.; Omengo, F. O.; Govers, G.; Bouillon, S.

    2015-08-01

    A significant amount of carbon is transported to the ocean as dissolved organic carbon (DOC) in rivers. During transport, it can be transformed through microbial consumption and photochemical oxidation. In dark incubation experiments with water from the Tana River, Kenya, we examined the consumption of DOC through microbial decomposition and the associated change in its carbon stable isotope composition (δ13C). In 15 of the 18 incubations, DOC concentrations decreased significantly by 10 to 60 %, with most of the decomposition taking place within the first 24-48 h. After 8 days, the remaining DOC was up to 3 ‰ more depleted in 13C compared with the initial pool, and the change in δ13C correlated strongly with the fraction of DOC remaining. We propose that the shift in δ13C is consistent with greater microbial lability of DOC originating from herbaceous C4 vegetation than DOC derived from woody C3 vegetation in the semi-arid lower Tana. The findings complement earlier data that riverine C sources do not necessarily reflect their proportion in the catchment: besides spatial distribution, also processing within the river can further influence the riverine δ13C.

  4. Dissolved organic carbon lability and stable isotope shifts during microbial decomposition in a tropical river system

    NASA Astrophysics Data System (ADS)

    Geeraert, N.; Omengo, F. O.; Govers, G.; Bouillon, S.

    2016-01-01

    A significant amount of carbon is transported to the ocean as dissolved organic carbon (DOC) in rivers. During transport, it can be transformed through microbial consumption and photochemical oxidation. In dark incubation experiments with water from the Tana River, Kenya, we examined the consumption of DOC through microbial decomposition and the associated change in its carbon stable isotope composition (δ13C). In 15 of the 18 incubations, DOC concentrations decreased significantly by 10 to 60 %, with most of the decomposition taking place within the first 24-48 h. After 8 days, the remaining DOC was up to 3 ‰ more depleted in 13C compared with the initial pool, and the change in δ13C correlated strongly with the fraction of DOC remaining. We hypothesize that the shift in δ13C is consistent with greater microbial lability of DOC originating from herbaceous C4 vegetation than DOC derived from woody C3 vegetation in the semi-arid lower Tana. The results complement earlier findings that the stable isotope concentration of riverine DOC does not necessarily reflect the proportion of C3 and C4-derived DOC in the catchment: besides spatial distribution patterns of different vegetation types, processing within the river can further influence the δ13C of riverine OC.

  5. An intermetallic Au24Ag20 superatom nanocluster stabilized by labile ligands.

    PubMed

    Wang, Yu; Su, Haifeng; Xu, Chaofa; Li, Gang; Gell, Lars; Lin, Shuichao; Tang, Zichao; Hkkinen, Hannu; Zheng, Nanfeng

    2015-04-01

    An intermetallic nanocluster containing 44 metal atoms, Au24Ag20(2-SPy)4(PhC?C)20Cl2, was successfully synthesized and structurally characterized by single-crystal analysis and density funtional theory computations. The 44 metal atoms in the cluster are arranged as a concentric three-shell Au12@Ag20@Au12 Keplerate structure having a high symmetry. For the first time, the co-presence of three different types of anionic ligands (i.e., phenylalkynyl, 2-pyridylthiolate, and chloride) was revealed on the surface of metal nanoclusters. Similar to thiolates, alkynyls bind linearly to surface Au atoms using their ?-bonds, leading to the formation of two types of surface staple units (PhC?C-Au-L, L = PhC?C(-) or 2-pyridylthiolate) on the cluster. The co-presence of three different surface ligands allows the site-specific surface and functional modification of the cluster. The lability of PhC?C(-) ligands on the cluster was demonstrated, making it possible to keep the metal core intact while removing partial surface capping. Moreover, it was found that ligand exchange on the cluster occurs easily to offer various derivatives with the same metal core but different surface functionality and thus different solubility. PMID:25803406

  6. A novel nanoparticulate system for sustained delivery of acid-labile lansoprazole.

    PubMed

    Alai, Milind Sadashiv; Lin, Wen Jen

    2013-11-01

    In the present study, an effort was made to develop the Eudragit RS100 based nanoparticulate system for sustained delivery of an acid-labile drug, lansoprazole (LPZ). LPZ-loaded Eudragit RS100 nanoparticles (ERSNPs) were prepared by oil-in-water emulsion-solvent evaporation method. The effects of various formulation variables such as polymer concentration, drug amount and solvent composition on physicochemical performance of nanoparticles and in vitro drug release were investigated. All nanoparticles were spherical with particle size 198.9 ± 8.6-376.9 ± 5.6 nm and zeta potential +35.1 ± 1.7 to +40.2 ± 0.8 mV. The yield of nanoparticles was unaffected by change of these three variables. However, the drug loading and encapsulation efficiency were affected by polymer concentration and drug amount. On the other hand, the particle size of nanoparticles was significantly affected by polymer concentration and internal phase composition due to influence of droplet size during emulsification process. All nanoparticles prolonged drug release for 24h which was dominated by a combination of drug diffusion and polymer chain relaxation. The fastest and the slowest release rates were observed in C2-1002-10/0 and C8-4001-10/0, respectively, based on the release rate constant (k). Thus, the developed nanoparticles possessed a potential as a nano-carrier to sustain drug delivery for treatment of acid related disorders. PMID:23867305

  7. Soft antimicrobial agents: synthesis and activity of labile environmentally friendly long chain quaternary ammonium compounds.

    PubMed

    Thorsteinsson, Thorsteinn; Msson, Mr; Kristinsson, Karl G; Hjlmarsdttir, Martha A; Hilmarsson, Hilmar; Loftsson, Thorsteinn

    2003-09-11

    A series of soft quaternary ammonium antimicrobial agents, which are analogues to currently used quaternary ammonium preservatives such as cetyl pyridinium chloride and benzalkonium chloride, were synthesized. These soft analogues consist of long alkyl chain connected to a polar headgroup via chemically labile spacer group. They are characterized by facile nonenzymatic and enzymatic degradation to form their original nontoxic building blocks. However, their chemical stability has to be adequate in order for them to have antimicrobial effects. Stability studies and antibacterial and antiviral activity measurements revealed relationship between activity, lipophilicity, and stability. Their minimum inhibitory concentration (MIC) was as low as 1 microg/mL, and their viral reduction was in some cases greater than 6.7 log. The structure-activity studies demonstrate that the bioactive compounds (i.e., MIC for Gram-positive bacteria of <10 microg/mL) have an alkyl chain length between 12 and 18 carbon atoms, with a polar headgroup preferably of a small quaternary ammonium group, and their acquired inactivation half-life must be greater than 3 h at 60 degrees C. PMID:12954069

  8. Determination of labile copper, cobalt, and chromium in textile mill wastewater

    SciTech Connect

    Crain, J.S.; Essling, A.M.; Kiely, J.T.

    1997-01-01

    Copper, chromium, and cobalt species present in filtered wastewater effluent were separated by cation exchange and reverse phase chromatography. Three sample fractions were obtained: one containing metal cations (i.e., trivalent Cr, divalent Cu, and divalent Co), one containing organic species (including metallized dyes), and one containing other unretained species. The metal content of each fraction was determined by inductively coupled plasma atomic emission spectroscopy (ICP-AES). The sum of the corrected data was compared to the metal content of a filtered effluent aliquot digested totally with fuming sulfuric acid. Other aliquots of the filtered effluent were spiked with the metals of interest and digested to confirm chemical yield and accuracy. Method detection limits were consistently below 20 {mu}g L{sup -1} for Cu, 30 {mu}g L{sup -1} for Co, and 10 {mu}g L{sup -1} for Cr. Spike recoveries for undifferentiated Cu and Cr were statistically indistinguishable from unity; although Co spike recoveries were slightly low ({approximately}95%), its chemical yield was 98%. Copper retention on the sodium sulfonate cation exchange resin was closely correlated with the [EDTA]/[Cu] ratio, suggesting that metals retained upon the cation exchange column were assignable to labile metal species; however, mass balances for all three elements, though reasonable ({approximately}90%), were significantly different from unity. Mechanical factors may have contributed to the material loss, but other data suggest that some metal species reacted irreversibly with the reverse phase column. 3 refs., 2 figs., 4 tabs.

  9. [Demonstration of a liposoluble ciguateric toxin in Caranx bartholomaei caught in the French West Indies].

    PubMed

    Vernoux, J P; Gaign, M; Riyeche, N; Tagmouti, F; Magras, L P; Nolen, J

    1982-10-01

    This report deals with some of the biological and chemical properties of a liposoluble poison extracted from Caranx bartholomaei (yellow jack), a toxic fish from the French West Indies (St Barth's island). Within a single fish, poison concentration is higher in the viscera; toxicity level is uniform throughout the flesh. In the few specimens tested, liver toxicity variations parallel those of flesh. The poison is heat stable so that cooking does not impair the toxicity. It is soluble in acetone, diethyl ether, chloroform, benzene, methanol and ethanol, but insoluble in n. hexane. Crude toxin injection or ingestion induces ciguateric disease in cats, mice or newly born chicken. Crude toxin stability is good after 30 minutes at 90 degrees C in a 0,5 N solution of a weak acid but not of a weak base. In the same conditions, loss of activity is nearly complete with a strong acid or a strong base. Moreover, rapid alkali treatment at room temperature destroys more than 50 per cent of the toxicity. The toxin is eluted by chloroform-methanol (9:1) from a silicic acid column and by acetone-methanol (9,5:0,5) from a Florisil column. DEAE cellulose column chromatography clearly separates the toxin into two lethal components; but thin layer chromatography of crude or fractionated toxin indicates only one toxic band in three different solvent systems. As a conclusion, in spite of a few differences related to instability in alkalin medium and elution from DEAE cellulose, this poison is quite similar to those carried by ciguatoxic fishes from the Pacific area. The differences noted above could be accounted for either by a specific metabolism of the species studied or by differences in the causative agent due to geographical location, or both. PMID:6817824

  10. Bacterial RTX Toxins Allow Acute ATP Release from Human Erythrocytes Directly through the Toxin Pore*

    PubMed Central

    Skals, Marianne; Bjaelde, Randi G.; Reinholdt, Jesper; Poulsen, Knud; Vad, Brian S.; Otzen, Daniel E.; Leipziger, Jens; Praetorius, Helle A.

    2014-01-01

    ATP is as an extracellular signaling molecule able to amplify the cell lysis inflicted by certain bacterial toxins including the two RTX toxins ?-hemolysin (HlyA) from Escherichia coli and leukotoxin A (LtxA) from Aggregatibacter actinomycetemcomitans. Inhibition of P2X receptors completely blocks the RTX toxin-induced hemolysis over a larger concentration range. It is, however, at present not known how the ATP that provides the amplification is released from the attacked cells. Here we show that both HlyA and LtxA trigger acute release of ATP from human erythrocytes that preceded and were not caused by cell lysis. This early ATP release did not occur via previously described ATP-release pathways in the erythrocyte. Both HlyA and LtxA were capable of triggering ATP release in the presence of the pannexin 1 blockers carbenoxolone and probenecid, and the HlyA-induced ATP release was found to be similar in erythrocytes from pannexin 1 wild type and knock-out mice. Moreover, the voltage-dependent anion channel antagonist TRO19622 had no effect on ATP release by either of the toxins. Finally, we showed that both HlyA and LtxA were able to release ATP from ATP-loaded lipid (1-palmitoyl-2-oleoyl-phosphatidylcholine) vesicles devoid of any erythrocyte channels or transporters. Again we were able to show that this happened in a non-lytic fashion, using calcein-containing vesicles as controls. These data show that both toxins incorporate into lipid vesicles and allow ATP to be released. We suggest that both toxins cause acute ATP release by letting ATP pass the toxin pores in both human erythrocytes and artificial membranes. PMID:24860098

  11. A new one-step method for the preparation of 3',5'-bisphosphates of acid-labile deoxynucleosides.

    PubMed

    Folkman, W; Kuśmierek, J T; Singer, B

    1990-01-01

    A method is described for preparing 3',5'-bisphosphates of labile deoxynucleosides. Under strictly anhydrous conditions, pyrophosphoryl tetrakistriazole apparently forms a ring structure bridging the 3'- and 5'-hydroxyl groups of deoxynucleosides, since upon the addition of water the ring opens and the 3',5'-bisphosphate is formed. Due to the presence of triethylamine no acid is generated at any time so that the entire procedure is in neutral solution. The bisphosphates of N2,3-ethenodeoxyguanosine, O2-ethyldeoxythymidine, and O4-methyldeoxythymidine, all of which are acid-labile, were prepared in good yield without degradation. Other modified bisphosphates prepared include O6-benzyldeoxyguanosine and 1,N6-ethenodeoxyadenosine, as well as those of unmodified deoxyguanosine and thymidine. Characterization was by 31P NMR and UV spectroscopy. Both 5'p(dT)p3' and 5'p(dG)p3' were substrates for RNA ligase, further proving the structure of the phosphorylated compounds. PMID:2103324

  12. Metabolically labile cannabinoid esters: a 'soft drug' approach for the development of cannabinoid-based therapeutic drugs.

    PubMed

    Minutolo, F; Cascio, M G; Carboni, I; Bisogno, T; Prota, G; Bertini, S; Digiacomo, M; Bifulco, M; Di Marzo, V; Macchia, M

    2007-09-01

    Biphenylic ester derivatives, designed by using a 'soft-drug' approach, proved to possess good binding properties toward cannabinoid CB(1) and CB(2) receptors and, at the same time, their metabolically labile ester portion would promote a rapid systemic inactivation. This may constitute a possible solution to the psychotropic side effects encountered when cannabinoids are therapeutically employed as local analgesic or antiglaucoma agents. PMID:17604171

  13. A study on the membrane depolarization of skeletal muscles caused by a scorpion toxin, sea anemone toxin II and crotamine and the interaction between toxins.

    PubMed Central

    Chang, C. C.; Hong, S. J.; Su, M. J.

    1983-01-01

    Quinquestriatus toxin (QTX) isolated from the venom of a scorpion (Leiurus quinquestriatus) and sea anemone (Anemonia sulcata) toxin II enhanced the twitch response of the rat and mouse diaphragms and like crotamine (isolated from the venom of Crotalus durissus terrificus) caused spontaneous fasciculation of the muscle. Trains of action potentials in muscles at 70-250 Hz, which could not be antagonized by (+)-tubocurarine, were triggered by single stimulation or occurred spontaneously after treatment with these toxins. QTX and toxin II prolonged the rat muscle action potential 3 to 4 fold whereas crotamine prolonged the action potential by only 30%. The membrane potential was depolarized from about -82 mV to -55 mV by crotamine 2 micrograms ml-1, -41 mV by toxin II 5 micrograms ml-1 and to -50 mV by QTX 1 microgram ml-1. The concentrations to induce 50% maximal depolarization (K0.5) were 0.07, 0.15 and greater than 0.4 microgram ml-1, respectively, for QTX, crotamine and toxin II, whereas the rates of depolarization were in the order toxin II greater than or equal to crotamine greater than QTX. The depolarizing effects of crotamine and QTX, but not of toxin II, were saturable. The depolarizing effects of all three toxins were irreversible whereas the membrane potential could be restored by tetrodotoxin non-competitively. Simultaneous treatment with crotamine and QTX or crotamine and toxin II at concentrations below K0.5 caused only additive effects on depolarization. When the muscle was depolarized by pretreating with a saturating concentration of crotamine, the onset of depolarization by QTX was greatly retarded whereas that by toxin II was unaffected.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:6197125

  14. Molecular Imaging of Labile Iron(II) Pools in Living Cells with a Turn-On Fluorescent Probe

    PubMed Central

    Au-Yeung, Ho Yu; Chan, Jefferson; Chantarojsiri, Teera; Chang, Christopher J.

    2013-01-01

    Iron is an essential metal for living organisms, but misregulation of its homeostasis at the cellular level can trigger detrimental oxidative and/or nitrosative stress and damage events. Motivated to help study the physiological and pathological consequences of biological iron regulation, we now report a reaction-based strategy for monitoring labile Fe2+ pools in aqueous solution and in living cells. Iron Probe 1 (IP1) exploits a bioinspired, iron-mediated oxidative C–O bond cleavage reaction to achieve a selective turn-on response to Fe2+ over a range of cellular metal ions in their bioavailable forms. We show that this first-generation chemical tool for fluorescence Fe2+ detection can visualize changes in exchangeable iron stores in living cells upon iron supplementation or depletion, including labile iron pools at endogenous, basal levels. Moreover, IP1 can be used to identify reversible expansion of labile iron pools by stimulation with vitamin C or the iron regulatory hormone hepcidin, providing a starting point for further investigations of iron signaling and stress events in living systems as well as future probe development. PMID:24063668

  15. Microwave-assisted extraction performed in low temperature and in vacuo for the extraction of labile compounds in food samples.

    PubMed

    Xiao, Xiaohua; Song, Wei; Wang, Jiayue; Li, Gongke

    2012-01-27

    In this study, low temperature vacuum microwave-assisted extraction, which simultaneous performed microwave-assisted extraction (MAE) in low temperature and in vacuo environment, was proposed. The influencing parameters including solid/liquid ratio, extraction temperature, extraction time, degree of vacuum and microwave power were discussed. The predominance of low temperature vacuum microwave-assisted extraction was investigated by comparing the extraction yields of vitamin C, ?-carotene, aloin A and astaxanthin in different foods with that in MAE and solvent extraction, and 5.2-243% increments were obtained. On the other hand, the chemical kinetics of vitamin C and aloin A, which composed two different steps including the extraction step of analyte transferred from matrix into solvent and the decomposition step of analyte degraded in the extraction solvent, were proposed. All of the decomposition rates (K(2)) for the selected analyte in low temperature, in vacuo and in nitrogen atmosphere decreased significantly comparing with that in conventional MAE, which are in agreement with that obtained from experiments. Consequently, the present method was successfully applied to extract labile compound from different food samples. These results showed that low temperature and/or in vacuo environment in microwave-assisted extraction system was especially important to prevent the degradation of labile components and have good potential on the extraction of labile compound in foods, pharmaceutical and natural products. PMID:22177069

  16. CD44 Binding to Hyaluronic Acid Is Redox Regulated by a Labile Disulfide Bond in the Hyaluronic Acid Binding Site

    PubMed Central

    Kellett-Clarke, Helena; Stegmann, Monika; Barclay, A. Neil; Metcalfe, Clive

    2015-01-01

    CD44 is the primary leukocyte cell surface receptor for hyaluronic acid (HA), a component of the extracellular matrix. Enzymatic post translational cleavage of labile disulfide bonds is a mechanism by which proteins are structurally regulated by imparting an allosteric change and altering activity. We have identified one such disulfide bond in CD44 formed by Cys77 and Cys97 that stabilises the HA binding groove. This bond is labile on the surface of leukocytes treated with chemical and enzymatic reducing agents. Analysis of CD44 crystal structures reveal the disulfide bond to be solvent accessible and in theLH hook configuration characteristic of labile disulfide bonds. Kinetic trapping and binding experiments on CD44-Fc chimeric proteins show the bond is preferentially reduced over the other disulfide bonds in CD44 and reduction inhibits the CD44-HA interaction. Furthermore cells transfected with CD44 no longer adhere to HA coated surfaces after pre-treatment with reducing agents. The implications of CD44 redox regulation are discussed in the context of immune function, disease and therapeutic strategies. PMID:26379032

  17. Visualization of Peroxynitrite-Induced Changes of Labile Zn2+ in the Endoplasmic Reticulum with Benzoresorufin-based Fluorescent Probes

    PubMed Central

    Lin, Wei; Buccella, Daniela; Lippard, Stephen J.

    2013-01-01