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Sample records for americanus induces neutralizing

  1. An enzymatically inactivated hemoglobinase from Necator americanus induces neutralizing antibodies against multiple hookworm species and protects dogs against heterologous hookworm infection

    PubMed Central

    Pearson, Mark S.; Bethony, Jeffrey M.; Pickering, Darren A.; de Oliveira, Luciana M.; Jariwala, Amar; Santiago, Helton; Miles, Aaron P.; Zhan, Bin; Jiang, Desheng; Ranjit, Najju; Mulvenna, Jason; Tribolet, Leon; Plieskatt, Jordan; Smith, Tracey; Bottazzi, Maria Elena; Jones, Kathryn; Keegan, Brian; Hotez, Peter J.; Loukas, Alex

    2009-01-01

    Hookworms digest hemoglobin from erythrocytes via a proteolytic cascade that begins with the aspartic protease, APR-1. Ac-APR-1 from the dog hookworm, Ancylostoma caninum, protects dogs against hookworm infection via antibodies that neutralize enzymatic activity and interrupt blood-feeding. Toward developing a human hookworm vaccine, we expressed both wild-type (Na-APR-1wt) and mutant (Na-APR-1mut—mutagenesis of the catalytic aspartic acids) forms of Na-APR-1 from the human hookworm, Necator americanus. Refolded Na-APR-1wt was catalytically active, and Na-APR-1mut was catalytically inactive but still bound substrates. Vaccination of canines with Na-APR-1mut and heterologous challenge with A. caninum resulted in significantly reduced parasite egg burdens (P=0.034) and weight loss (P=0.022). Vaccinated dogs also had less gut pathology, fewer adult worms, and reduced blood loss compared to controls but these did not reach statistical significance. Vaccination with Na-APR-1mut induced antibodies that bound the native enzyme in the parasite gut and neutralized enzymatic activity of Na-APR-1wt and APR-1 orthologues from three other hookworm species that infect humans. IgG1 against Na-APR-1mut was the most prominently detected antibody in sera from people resident in high-transmission areas for N. americanus, indicating that natural boosting may occur in exposed humans. Na-APR-1mut is now a lead antigen for the development of an antihematophagy vaccine for human hookworm disease.—Pearson, M. S., Bethony, J. M., Pickering, D. A., de Oliveira, L. M., Jariwala, A., Santiago, H., Miles, A. P., Zhan, B., Jiang, D., Ranjit, N., Mulvenna, J., Tribolet, L., Plieskatt, J., Smith, T., Bottazzi, M. E., Jones, K., Keegan, B., Hotez, P. J., Loukas, A. An enzymatically inactivated hemoglobinase from Necator americanus induces neutralizing antibodies against multiple hookworm species and protects dogs against heterologous hookworm infection. PMID:19380510

  2. An enzymatically inactivated hemoglobinase from Necator americanus induces neutralizing antibodies against multiple hookworm species and protects dogs against heterologous hookworm infection.

    PubMed

    Pearson, Mark S; Bethony, Jeffrey M; Pickering, Darren A; de Oliveira, Luciana M; Jariwala, Amar; Santiago, Helton; Miles, Aaron P; Zhan, Bin; Jiang, Desheng; Ranjit, Najju; Mulvenna, Jason; Tribolet, Leon; Plieskatt, Jordan; Smith, Tracey; Bottazzi, Maria Elena; Jones, Kathryn; Keegan, Brian; Hotez, Peter J; Loukas, Alex

    2009-09-01

    Hookworms digest hemoglobin from erythrocytes via a proteolytic cascade that begins with the aspartic protease, APR-1. Ac-APR-1 from the dog hookworm, Ancylostoma caninum, protects dogs against hookworm infection via antibodies that neutralize enzymatic activity and interrupt blood-feeding. Toward developing a human hookworm vaccine, we expressed both wild-type (Na-APR-1(wt)) and mutant (Na-APR-1(mut)-mutagenesis of the catalytic aspartic acids) forms of Na-APR-1 from the human hookworm, Necator americanus. Refolded Na-APR-1(wt) was catalytically active, and Na-APR-1(mut) was catalytically inactive but still bound substrates. Vaccination of canines with Na-APR-1(mut) and heterologous challenge with A. caninum resulted in significantly reduced parasite egg burdens (P=0.034) and weight loss (P=0.022). Vaccinated dogs also had less gut pathology, fewer adult worms, and reduced blood loss compared to controls but these did not reach statistical significance. Vaccination with Na-APR-1(mut) induced antibodies that bound the native enzyme in the parasite gut and neutralized enzymatic activity of Na-APR-1(wt) and APR-1 orthologues from three other hookworm species that infect humans. IgG1 against Na-APR-1(mut) was the most prominently detected antibody in sera from people resident in high-transmission areas for N. americanus, indicating that natural boosting may occur in exposed humans. Na-APR-1(mut) is now a lead antigen for the development of an antihematophagy vaccine for human hookworm disease.

  3. Esfenvalerate-induced case-abandonment in the larvae of the caddisfly (Brachycentrus americanus).

    PubMed

    Johnson, Katherine R; Jepson, Paul C; Jenkins, Jeffrey J

    2008-02-01

    Field-collected Brachycentrus americanus Banks (Trichoptera: Brachycentridae) larvae were used to investigate the relationship between esfenvalerate exposure and case-abandonment response, determine larval ability to construct a new case, and measure the change in predation risk to insects in rebuilt cases. We evaluated case-abandonment following four environmentally relevant esfenvalerate exposures, 0.05, 0.1, 0.2, and 0.4 microg/L; 48-h exposures to 0.2 and 0.4 microg/L (nominal) esfenvalerate both resulted in over 60% of larvae abandoning cases and were statistically indistinguishable. Propensity to engage in building behaviors was significantly diminished in 0.2 and 0.4 microg/L esfenvalerate-exposed insects that had abandoned cases, with less than 20% of exposed insects producing cases. Cases built by intoxicated larvae were characterized by a disorganized composition, and required half the pressure to crush versus cases built by nonexposed larvae. Pre-exposing case-building material to 1 microg/L esfenvalerate also reduced the physical strength of rebuilt cases. Larvae inhabiting weaker rebuilt cases and larvae without cases were significantly more susceptible to predation by second year Hesperoperla pacifica Banks (Plecoptera: Perlidae) stonefly nymphs than those in original cases. Overall, we concluded that small behavioral responses can have profound consequences for survival of species and reveal susceptible stages in life-cycles that can be overlooked by conventional approaches to ecological risk assessment.

  4. Assessment of Dredge-induced Sedimentation Effects on Winter Flounder (pseudopleuronectes americanus) Hatching Success: Results of Laboratory Investigations

    EPA Science Inventory

    Historically of significant commercial and recreational value, winter flounder (Pseudopleuronectes americanus) stocks have declined steadily over the last 20 years and are currently at an all-time low. Although overfishing and climate change have been implicated, habitat alterat...

  5. Assessment of Dredging-Induced Sedimentation Effects on Winter Flounder (Pseudopleuronectes americanus) Hatching Success: Results of Laboratory Investigations

    EPA Science Inventory

    Historically of significant commercial and recreational value, winter flounder (Pseudopleuronectes americanus) stocks have declined steadily over the last 20 years and are currently at an all-time low. Although overfishing and climate change have been implicated, habitat alterat...

  6. Assessment of Dredge-induced Sedimentation Effects on Winter Flounder (pseudopleuronectes americanus) Hatching Success: Results of Laboratory Investigations

    EPA Science Inventory

    Historically of significant commercial and recreational value, winter flounder (Pseudopleuronectes americanus) stocks have declined steadily over the last 20 years and are currently at an all-time low. Although overfishing and climate change have been implicated, habitat alterat...

  7. Assessment of Dredging-Induced Sedimentation Effects on Winter Flounder (Pseudopleuronectes americanus) Hatching Success: Results of Laboratory Investigations

    EPA Science Inventory

    Historically of significant commercial and recreational value, winter flounder (Pseudopleuronectes americanus) stocks have declined steadily over the last 20 years and are currently at an all-time low. Although overfishing and climate change have been implicated, habitat alterat...

  8. Candidatus Liberibacter americanus induces significant reprogramming of the transcriptome of the susceptible citrus genotype

    PubMed Central

    2013-01-01

    Background Citrus huanglongbing (HLB) disease is caused by endogenous, phloem-restricted, Gram negative, uncultured bacteria named Candidatus Liberibacter africanus (CaLaf), Ca. L. asiaticus (CaLas), and Ca. L. americanus (CaLam), depending on the continent where the bacteria were first detected. The Asian citrus psyllid vector, Diaphorina citri, transmits CaLas and CaLam and both Liberibacter species are present in Brazil. Several studies of the transcriptional response of citrus plants manifesting HLB symptoms have been reported, but only for CaLas infection. This study evaluated the transcriptional reprogramming of a susceptible genotype of sweet orange challenged with CaLam, using a customized 385K microarray containing approximately 32,000 unigene transcripts. We analyzed global changes in gene expression of CaLam-infected leaves of sweet orange during the symptomatic stage of infection and compared the results with previously published microarray studies that used CaLas-infected plants. Twenty candidate genes were selected to validate the expression profiles in symptomatic and asymptomatic PCR-positive leaves infected with CaLas or CaLam. Results The microarray analysis identified 633 differentially expressed genes during the symptomatic stage of CaLam infection. Among them, 418 (66%) were upregulated and 215 (34%) were down regulated. Five hundred and fourteen genes (81%) were orthologs of genes from Arabidopsis thaliana. Gene set enrichment analysis (GSEA) revealed that several of the transcripts encoded transporters associated with the endomembrane system, especially zinc transport. Among the most biologically relevant gene transcripts in GSEA were those related to signaling, metabolism and/or stimulus to hormones, genes responding to stress and pathogenesis, biosynthesis of secondary metabolites, oxidative stress and transcription factors belonging to different families. Real time PCR of 20 candidate genes validated the expression pattern of some genes in

  9. Eastern equine encephalitis in moose (Alces americanus) in northeastern Vermont.

    PubMed

    Mutebi, John-Paul; Swope, Bethany N; Saxton-Shaw, Kali D; Graham, Alan C; Turmel, Jon P; Berl, Erica

    2012-10-01

    During fall 2010, 21 moose (Alces americanus) sera collected in northeastern Vermont were screened for eastern equine encephalitis virus (EEEV) antibodies using plaque reduction neutralization tests. Six (29%) were antibody positive. This is the first evidence of EEEV activity in Vermont, and the second report of EEEV antibodies in moose.

  10. Neutralizing antibodies to HIV-1 induced by immunization

    PubMed Central

    McCoy, Laura E.

    2013-01-01

    Most neutralizing antibodies act at the earliest steps of viral infection and block interaction of the virus with cellular receptors to prevent entry into host cells. The inability to induce neutralizing antibodies to HIV has been a major obstacle to HIV vaccine research since the early days of the epidemic. However, in the past three years, the definition of a neutralizing antibody against HIV has been revolutionized by the isolation of extremely broad and potent neutralizing antibodies from HIV-infected individuals. Considerable hurdles remain for inducing neutralizing antibodies to a protective level after immunization. Meanwhile, novel technologies to bypass the induction of antibodies are being explored to provide prophylactic antibody-based interventions. This review addresses the challenge of inducing HIV neutralizing antibodies upon immunization and considers notable recent advances in the field. A greater understanding of the successes and failures for inducing a neutralizing response upon immunization is required to accelerate the development of an effective HIV vaccine. PMID:23401570

  11. Necator americanus: The Na-ASP-2 protein secreted by the infective larvae induces neutrophil recruitment in vivo and in vitro

    PubMed Central

    Bower, Molly A.; Constant, Stephanie L.; Mendez, Susana

    2008-01-01

    The L3-secreted Ancylostoma Secreted Protein-2 from the human hookworm Necator americanus (Na-ASP-2) has been selected as a candidate vaccine antigen in anticipation of clinical trials. Its crystal structure revealed that Na-ASP-2 has structural and charge similarities to CC-chemokines, suggesting that it might act as a chemokine mimic when released by the infective larvae during tissue migration. Using the air pouch model of acute inflammation, we found that Na-ASP-2 induced a significant leukocyte influx to the skin pouch, mostly comprised of neutrophils (60%) and monocytes (30%) that was transient and resolved in 24 h. Other hookworm larval proteins did not cause any inflammatory leukocytes to migrate into air pouches. In vitro chemotaxis assays confirmed our results and demonstrated that leukocyte migration was a direct effect of Na-ASP-2 exposure and not caused by other molecules released by host cells in the inflammatory microenvironment or by the expression vector. PMID:18199436

  12. The Relationship between Neutralization Techniques and Induced Abortion.

    PubMed

    Kalateh Sadati, Ahmad; Tabei, Seyed Ziaaddin; Salehzadeh, Hamzeh; Rahnavard, Farnaz; Namavar Jahromi, Bahia; Hemmati, Soroor

    2014-04-01

    Induced abortion is not only a serious threat for women's health, but also a controversial topic for its ethical and moral problems. We aimed to evaluate the relationship between neutralization techniques and attempting to commit abortion in married women with unintended pregnancy. After in-depth interviews with some women who had attempted abortion, neutralization themes were gathered. Next, to analyze the data quantitatively, a questionnaire was created including demographic and psychosocial variables specifically related to neutralization. The participants were divided into two groups (abortion and control) of unintended pregnancy and were then compared. Analysis of psychosocial variables revealed a significant difference in the two groups at neutralization, showing that neutralization in the control group (56.97±10.24) was higher than that in the abortion group (44.19±12.44). To evaluate the findings more accurately, we examined the causal factors behind the behaviors of the abortion group. Binary logistic regression showed that among psychosocial factors, neutralization significantly affected abortion (95% CI=1.07-1.35). Despite the network of many factors affecting induced abortion, neutralization plays an important role in reinforcing the tendency to attempt abortion. Furthermore, the decline of religious beliefs, as a result of the secular context of the modern world, seems to have an important role in neutralizing induced abortion.

  13. The Relationship between Neutralization Techniques and Induced Abortion

    PubMed Central

    Kalateh Sadati, Ahmad; Tabei, Seyed Ziaaddin; Salehzadeh, Hamzeh; Rahnavard, Farnaz; Namavar Jahromi, Bahia; Hemmati, Soroor

    2014-01-01

    Background: Induced abortion is not only a serious threat for women’s health, but also a controversial topic for its ethical and moral problems. We aimed to evaluate the relationship between neutralization techniques and attempting to commit abortion in married women with unintended pregnancy. Methods: After in-depth interviews with some women who had attempted abortion, neutralization themes were gathered. Next, to analyze the data quantitatively, a questionnaire was created including demographic and psychosocial variables specifically related to neutralization. The participants were divided into two groups (abortion and control) of unintended pregnancy and were then compared. Results: Analysis of psychosocial variables revealed a significant difference in the two groups at neutralization, showing that neutralization in the control group (56.97±10.24) was higher than that in the abortion group (44.19±12.44). To evaluate the findings more accurately, we examined the causal factors behind the behaviors of the abortion group. Binary logistic regression showed that among psychosocial factors, neutralization significantly affected abortion (95% CI=1.07-1.35). Conclusion: Despite the network of many factors affecting induced abortion, neutralization plays an important role in reinforcing the tendency to attempt abortion. Furthermore, the decline of religious beliefs, as a result of the secular context of the modern world, seems to have an important role in neutralizing induced abortion. PMID:25349851

  14. HIV Neutralizing Antibodies Induced by Native-like Envelope Trimers

    PubMed Central

    Sanders, Rogier W.; van Gils, Marit J.; Derking, Ronald; Sok, Devin; Ketas, Thomas J.; Burger, Judith A.; Ozorowski, Gabriel; Cupo, Albert; Simonich, Cassandra; Goo, Leslie; Arendt, Heather; Kim, Helen J.; Lee, Jeong Hyun; Pugach, Pavel; Williams, Melissa; Debnath, Gargi; Moldt, Brian; van Breemen, Mariëlle J.; Isik, Gözde; Medina-Ramírez, Max; Back, Jaap Willem; Koff, Wayne; Julien, Jean-Philippe; Rakasz, Eva G.; Seaman, Michael S.; Guttman, Miklos; Lee, Kelly K.; Klasse, Per Johan; LaBranche, Celia; Schief, William R.; Wilson, Ian A.; Overbaugh, Julie; Burton, Dennis R.; Ward, Andrew B.; Montefiori, David C.; Dean, Hansi; Moore, John P.

    2015-01-01

    A challenge for HIV-1 immunogen design is inducing neutralizing antibodies (NAbs) against neutralization-resistant (Tier-2) viruses that dominate human transmissions. We show that a soluble recombinant HIV-1 envelope glycoprotein trimer that adopts a native conformation (BG505 SOSIP.664) induced NAbs potently against the sequence-matched Tier-2 virus in rabbits and similar but weaker responses in macaques. The trimer also consistently induced cross-reactive NAbs against more sensitive (Tier-1) viruses. Tier-2 NAbs recognized conformational epitopes that differed between animals and in some cases overlapped with those recognized by broadly neutralizing antibodies (bNAbs), whereas Tier-1 responses targeted linear V3 epitopes. A second trimer, B41 SOSIP.664, also induced a strong autologous Tier-2 NAb response in rabbits. Thus, native-like trimers represent a promising starting point for developing HIV-1 vaccines aimed at inducing bNAbs. PMID:26089353

  15. Cross-clade neutralizing antibodies against HIV-1 induced in rabbits by focusing the immune response on a neutralizing epitope

    SciTech Connect

    Zolla-Pazner, Susan; Cohen, Sandra; Pinter, Abraham; Krachmarov, Chavdar; Wrin, Terri; Wang Shixia; Lu Shan

    2009-09-15

    Studies were performed to induce cross-clade neutralizing antibodies (Abs) by testing various combinations of prime and boost constructs that focus the immune response on structurally-conserved epitopes in the V3 loop of HIV-1 gp120. Rabbits were immunized with gp120 DNA containing a V3 loop characterized by the GPGR motif at its tip, and/or with gp120 DNA with a V3 loop carrying the GPGQ motif. Priming was followed by boosts with V3-fusion proteins (V3-FPs) carrying the V3 sequence from a subtype B virus (GPGR motif), and/or with V3 sequences from subtypes A and C (GPGQ motif). The broadest and most consistent neutralizing responses were generated when using a clade C gp120 DNA prime and with the V3{sub B}-FP boost. Immune sera displayed neutralizing activity in three assays against pseudoviruses and primary isolates from subtypes A, AG, B, C, and D. Polyclonal Abs in the immune rabbit sera neutralized viruses that were not neutralized by pools of human anti-V3 monoclonal Abs. Greater than 80% of the neutralizing Abs were specific for V3, showing that the immune response could be focused on a neutralizing epitope and that vaccine-induced anti-V3 Abs have cross-clade neutralizing activity.

  16. Hepatitis C virus vaccine candidates inducing protective neutralizing antibodies.

    PubMed

    Fauvelle, Catherine; Colpitts, Che C; Keck, Zhen-Yong; Pierce, Brian G; Foung, Steven K H; Baumert, Thomas F

    2016-12-01

    With more than 150 million chronically infected people, hepatitis C virus (HCV) remains a substantial global health burden. Direct-acting antivirals have dramatically improved viral cure. However, limited access to therapy, late stage detection of infection and re-infection following cure illustrate the need for a vaccine for global control of infection. Vaccines with induction of neutralizing antibodies (nAbs) have been shown to protect successfully against infections by multiple viruses and are currently developed for HCV. Areas covered: Here we review the progress towards the development of vaccines aiming to confer protection against chronic HCV infection by inducing broadly nAbs. The understanding or viral immune evasion in infected patients, the development of novel model systems and the recent structural characterization of viral envelope glycoprotein E2 has markedly advanced our understanding of the molecular mechanisms of virus neutralization with the concomitant development of several vaccine candidates. Expert commentary: While HCV vaccine development remains challenged by the high viral diversity and immune evasion, marked progress in HCV research has advanced vaccine design. Several vaccine candidates have shown robust induction of nAbs in animal models and humans. Randomized clinical trials are the next step to assess their clinical efficacy for protection against chronic infection.

  17. CHARACTERIZATION OF MICROSATELLITE LOCI IN SCHOENOPLECTUS AMERICANUS (CYPERACEAE)

    EPA Science Inventory

    Schoenoplectus americanus is a model organism for studying ecological and ecosystem responses of salt marsh plant communities to global climate change. Here we characterize 16 microsatellite loci in S. americanus to facilitate studies on the genetic basis of phenotypic responses...

  18. Genome of the human hookworm Necator americanus.

    PubMed

    Tang, Yat T; Gao, Xin; Rosa, Bruce A; Abubucker, Sahar; Hallsworth-Pepin, Kymberlie; Martin, John; Tyagi, Rahul; Heizer, Esley; Zhang, Xu; Bhonagiri-Palsikar, Veena; Minx, Patrick; Warren, Wesley C; Wang, Qi; Zhan, Bin; Hotez, Peter J; Sternberg, Paul W; Dougall, Annette; Gaze, Soraya Torres; Mulvenna, Jason; Sotillo, Javier; Ranganathan, Shoba; Rabelo, Elida M; Wilson, Richard K; Felgner, Philip L; Bethony, Jeffrey; Hawdon, John M; Gasser, Robin B; Loukas, Alex; Mitreva, Makedonka

    2014-03-01

    The hookworm Necator americanus is the predominant soil-transmitted human parasite. Adult worms feed on blood in the small intestine, causing iron-deficiency anemia, malnutrition, growth and development stunting in children, and severe morbidity and mortality during pregnancy in women. We report sequencing and assembly of the N. americanus genome (244 Mb, 19,151 genes). Characterization of this first hookworm genome sequence identified genes orchestrating the hookworm's invasion of the human host, genes involved in blood feeding and development, and genes encoding proteins that represent new potential drug targets against hookworms. N. americanus has undergone a considerable and unique expansion of immunomodulator proteins, some of which we highlight as potential treatments against inflammatory diseases. We also used a protein microarray to demonstrate a postgenomic application of the hookworm genome sequence. This genome provides an invaluable resource to boost ongoing efforts toward fundamental and applied postgenomic research, including the development of new methods to control hookworm and human immunological diseases.

  19. HIV-1 envelope glycoprotein immunogens to induce broadly neutralizing antibodies.

    PubMed

    Sliepen, Kwinten; Sanders, Rogier W

    2016-01-01

    The long pursuit for a vaccine against human immunodeficiency virus 1 (HIV-1) has recently been boosted by a number of exciting developments. An HIV-1 subunit vaccine ideally should elicit potent broadly neutralizing antibodies (bNAbs), but raising bNAbs by vaccination has proved extremely difficult because of the characteristics of the HIV-1 envelope glycoprotein complex (Env). However, the isolation of bNAbs from HIV-1-infected patients demonstrates that the human humoral immune system is capable of making such antibodies. Therefore, a focus of HIV-1 vaccinology is the elicitation of bNAbs by engineered immunogens and by using vaccination strategies aimed at mimicking the bNAb maturation pathways in HIV-infected patients. Important clues can also be taken from the successful subunit vaccines against hepatitis B virus and human papillomavirus. Here, we review the different types of HIV-1 immunogens and vaccination strategies that are being explored in the search for an HIV-1 vaccine that induces bNAbs.

  20. MHD Induced Neutral Beam Ion Loss from NSTX Plasmas

    SciTech Connect

    D.S. Darrow, E.D. Fredrickson, N.N. Gorelenkov, A.L. Roquemore, and K. Shinohara

    2007-12-13

    Bursts of ~60 kHz activity on Mirnov coils occur frequently in NSTX plasmas and these are accompanied by bursts of neutral beam ion loss over a range in pitch angles. These losses have been measured with a scintillator type loss probe imaged with a high speed (>10,000 frames/s) video camera, giving the evolution of the energy and pitch angle distributions of the lost neutral beam ions over the course of the events. The instability occurs below the TAE frequency in NSTX (~100 kHz) in high beta plasmas and may be a beta driven Alfvén acoustic (BAAE) mode.

  1. Genome of the human hookworm Necator americanus

    PubMed Central

    Abubucker, Sahar; Hallsworth-Pepin, Kymberlie; Martin, John; Tyagi, Rahul; Heizer, Esley; Zhang, Xu; Bhonagiri-Palsikar, Veena; Minx, Patrick; Warren, Wesley C.; Wang, Qi; Zhan, Bin; Hotez, Peter J.; Sternberg, Paul W.; Dougall, Annette; Gaze, Soraya Torres; Mulvenna, Jason; Sotillo, Javier; Ranganathan, Shoba; Rabelo, Elida M.; Wilson, Richard W.; Felgner, Philip L.; Bethony, Jeffrey; Hawdon, John M.; Gasser, Robin B.; Loukas, Alex; Mitreva, Makedonka

    2014-01-01

    The hookworm Necator americanus is the predominant soil-transmitted human parasite. Adult worms feed on blood in the small intestine, causing iron deficiency anaemia, malnutrition, growth and development stunting in children, and severe morbidity and mortality during pregnancy in women. Characterization of the first hookworm genome sequence (244 Mb, 19,151 genes) identified genes orchestrating the hookworm's invasion of the human host, genes involved in blood feeding and development, and genes encoding proteins that represent new potential drug targets against hookworms. N. americanus has undergone a considerable and unique expansion of immunomodulator proteins, some of which we highlight as potential novel treatments against inflammatory diseases. We also utilize a protein microarray to demonstrate a post-genomic application of the hookworm genome sequence. This genome provides an invaluable resource to boost ongoing efforts towards fundamental and applied post-genomic research, including the development of new methods to control hookworm and human immunological diseases. PMID:24441737

  2. A pilot study on an attenuated Chinese EIAV vaccine inducing broadly neutralizing antibodies.

    PubMed

    Meng, Qinglai; Lin, Yuezhi; Ma, Jian; Ma, Yan; Zhao, Liping; Li, Shenwei; Liang, Hua; Zhou, Jianhua; Shen, Rongxian; Zhang, Xiaoyan; Shao, Yiming

    2011-08-01

    The attenuated Chinese equine infectious anemia virus (EIAV) vaccine has successfully protected millions of equine animals from EIA disease in China. In this pilot study, to determine whether this attenuated vaccine can induce broadly neutralizing antibodies, we immunized four horses with the attenuated Chinese vaccine strain EIAVFDDV and then observed the evolution of neutralizing antibodies against different EIAV strains. During the vaccination phase, all vaccinees rapidly developed high levels of neutralizing antibodies against the homologous vaccine strain (pLGFD3V), and 3 out of 4 horses showed a gradual increase in serum neutralizing activity against two relatively heterologous virulent variants of the challenge strain (pLGFD3Mu12V and DLV34). After challenge, the three horses that had developed high levels of neutralizing antibodies against pLGFD3Mu12V and DLV34 did not show signs of infection, which was demonstrated by immune suppression, while the one horse producing serum that could only neutralize pLGFD3V developed a febrile episode during the 8-month observation period. To assess whether the broadly neutralizing activity is associated with immune protection, sera drawn on the day of challenge from these four vaccinees and an additional four EIAVFDDV-vaccinated horses were analyzed for neutralizing antibodies against pLGFD3V, pLGFD3Mu12V and DLV34. Although there was no significant correlation between protection from infection and serum neutralizing activity against any of these three viral strains, protection from infection was observed to correlate better with serum neutralizing activity against the two heterologous virulent strains than against the homologous vaccine strain. These data indicate that EIAVFDDV induced broadly neutralizing antibodies, which might confer enhanced protection of vaccinees from infection by the challenge virus.

  3. [Induce of laccase from Trametes gallica and its degradation on neutral dyes and organophosphorus pesticides].

    PubMed

    Jing, De-Jun; Huang, Jian-Bo; Yang, Zhou-Ping; Hu, Rong; Cheng, Zi-Zhang; Huang, Qian-Ming

    2011-12-01

    The characteristics of the induction of laccase in Trametes gallica under different initial cultural pH, incubation time by different inducers were discussed, as well as the effects of temperature, pH and time on laccase degradation of six dyes and four organophosphors. The results showed that RB-bright blue, ABTS and o-toluidine affected the production of laccase at different levels, and ABTS was the best inductive agent in our test conditions, whose optimal initial pH and incubation time were 4.0 and 13 days, respectively. The appropriate reaction temperature of the laccase produced was 38 degrees C, and it got a good stability, for it could retain 78.6% of the enzyme activity after 20 min holding at 40 degrees C. Mediated by ABTS, the optimal temperature for laccase to degrade the six types of neutral dyes could be divided into two cases, that was 30 degrees C (neutral black, neutral bordeaux, neutral pink, methyl orange) and 60 degrees C (neutral dark yellow, cresol red), the optimal pH were 6.0 (neutral black), 2.0 (neutral bordeaux, neutral pink) and 4.0 (methyl orange, neutral dark yellow, cresol red), respectively, while the optimal times separately were 6 h (methyl orange, neutral dark yellow, cresol red), 12 h (neutral pink) and 24 h (neutral bordeaux). And using the same inductive agent, the best temperature for laccase to degrade dimethoate, chlorpyrifos, trichlorfon and parathion-pyridazine was 25 degrees C, the suitable time was 9 h, and the optimal pH was 10.0 for dimethoate, chlorpyrifos and parathion-pyridazine, and 8.0 for trichlorfon.

  4. HIV-1 VACCINES. HIV-1 neutralizing antibodies induced by native-like envelope trimers.

    PubMed

    Sanders, Rogier W; van Gils, Marit J; Derking, Ronald; Sok, Devin; Ketas, Thomas J; Burger, Judith A; Ozorowski, Gabriel; Cupo, Albert; Simonich, Cassandra; Goo, Leslie; Arendt, Heather; Kim, Helen J; Lee, Jeong Hyun; Pugach, Pavel; Williams, Melissa; Debnath, Gargi; Moldt, Brian; van Breemen, Mariëlle J; Isik, Gözde; Medina-Ramírez, Max; Back, Jaap Willem; Koff, Wayne C; Julien, Jean-Philippe; Rakasz, Eva G; Seaman, Michael S; Guttman, Miklos; Lee, Kelly K; Klasse, Per Johan; LaBranche, Celia; Schief, William R; Wilson, Ian A; Overbaugh, Julie; Burton, Dennis R; Ward, Andrew B; Montefiori, David C; Dean, Hansi; Moore, John P

    2015-07-10

    A challenge for HIV-1 immunogen design is the difficulty of inducing neutralizing antibodies (NAbs) against neutralization-resistant (tier 2) viruses that dominate human transmissions. We show that a soluble recombinant HIV-1 envelope glycoprotein trimer that adopts a native conformation, BG505 SOSIP.664, induced NAbs potently against the sequence-matched tier 2 virus in rabbits and similar but weaker responses in macaques. The trimer also consistently induced cross-reactive NAbs against more sensitive (tier 1) viruses. Tier 2 NAbs recognized conformational epitopes that differed between animals and in some cases overlapped with those recognized by broadly neutralizing antibodies (bNAbs), whereas tier 1 responses targeted linear V3 epitopes. A second trimer, B41 SOSIP.664, also induced a strong autologous tier 2 NAb response in rabbits. Thus, native-like trimers represent a promising starting point for the development of HIV-1 vaccines aimed at inducing bNAbs.

  5. Large-Scale Examination of Factors Influencing Phosphopeptide Neutral Loss during Collision Induced Dissociation

    PubMed Central

    Brown, Robert; Stuart, Scott A.; Houel, Stephane; Ahn, Natalie G.; Old, William M.

    2015-01-01

    Collision-induced dissociation (CID) remains the predominant mass spectrometry based method for identifying phosphorylation sites in complex mixtures. Unfortunately, the gas-phase reactivity of phosphoester bonds results in MS/MS spectra dominated by phosphoric acid (H3PO4) neutral loss events, suppressing informative peptide backbone cleavages. To understand the major drivers of H3PO4 neutral loss, we performed robust non-parametric statistical analysis of local and distal sequence effects on the magnitude and variability of neutral loss, using a collection of over 35,000 unique phosphopeptide MS/MS spectra. In contrast to peptide amide dissociation pathways, which are strongly influenced by adjacent amino acid side chains, we find that neutral loss of H3PO4 is affected by both proximal and distal sites, most notably basic residues and the peptide N-terminal primary amine. Previous studies have suggested that protonated basic residues catalyze neutral loss through direct interactions with the phosphate. In contrast, we find that nearby basic groups decrease neutral loss regardless of mobility class, an effect only seen by stratifying spectra by charge-mobility. The most inhibitory bases are those immediately N-terminal to the phosphate, presumably due to steric hindrances in catalyzing neutral loss. Further evidence of steric effects is shown by the presence of proline which can dramatically reduce the presence of neutral loss when between the phosphate and a possible charge donor. In mobile proton spectra the N-terminus is the strongest predictor of high neutral loss, with proximity to the N-terminus essential for peptides to exhibit the highest levels of neutral loss. PMID:25851653

  6. Ipomoea asarifolia neutralizes inflammation induced by Tityus serrulatus scorpion venom.

    PubMed

    Lima, Maira Conceição Jerônimo de Souza; Bitencourt, Mariana Angélica Oliveira; Furtado, Allanny Alves; Oliveira Rocha, Hugo Alexandre; Oliveira, Ruth Medeiros; da Silva-Júnior, Arnóbio Antônio; Tabosa do Egito, Eryvaldo Sócrates; Tambourgi, Denise Vilarinho; Zucolotto, Silvana Maria; Fernandes-Pedrosa, Matheus de Freitas

    2014-05-14

    Envenoming caused by scorpion sting is a serious public health problem. In Brazil, 13,038 accidents caused by venomous animals have been reported. Of this total, 53% of the cases and 14 deaths were caused by scorpions. Furthermore, Tityus serrulatus (Buthidae) is the most dangerous scorpion due to the high toxicity of its venom. The treatment is the common supportive therapy and the serum therapy, but some people do not have access to both therapies and seek healing through the use of medical plants. This study evaluated the ability of the crude extract and fractions from the leaves of Ipomoea asarifolia in neutralizing the main biological effects caused by Tityus serrulatus envenoming in mice. BALB/c mice were pretreated (i.v.) with 100 μλ of aqueous extracts and fractions dichloromethane, ethyl acetate, and n-butanol (CH₂Cl₂, EtOAc, and n-BuOH, respectively) of Ipomoea asarifolia, rutin or saline. Then, the animals received 100 μλ (i.p.) of venom of Tityus serrulatus (0.8 mg/kg). After six hours, the peritoneal lavage was performed with PBS and the number cells were determined using a Neubauer chamber. The supernatants were collected for determination of cytokines, such as IL-6, IL-12, and IL-1β. The aqueous extract, fractions and rutin, at all doses, significantly reduced cell migration, which was endorsed by the reduction of the levels of certain cytokines. This is the first study that demonstrated the potential effect of Ipomoea asarifolia against inflammation caused by Tityus serrulatus venom, suggesting that these extracts and/or their bioactive molecules, especially the flavonoid rutin, have potential use in the therapy of this envenomation. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  7. Ability of vaccine strain induced antibodies to neutralize field isolates of caliciviruses from Swedish cats.

    PubMed

    Wensman, Jonas Johansson; Samman, Ayman; Lindhe, Anna; Thibault, Jean-Christophe; Berndtsson, Louise Treiberg; Hosie, Margaret J

    2015-12-12

    Feline calicivirus (FCV) is a common cause of upper respiratory tract disease in cats worldwide. Its characteristically high mutation rate leads to escape from the humoral immune response induced by natural infection and/or vaccination and consequently vaccines are not always effective against field isolates. Thus, there is a need to continuously investigate the ability of FCV vaccine strain-induced antibodies to neutralize field isolates. Seventy-eight field isolates of FCV isolated during the years 2008-2012 from Swedish cats displaying clinical signs of upper respiratory tract disease were examined in this study. The field isolates were tested for cross-neutralization using a panel of eight anti-sera raised in four pairs of cats following infection with four vaccine strains (F9, 255, G1 and 431). The anti-sera raised against F9 and 255 neutralised 20.5 and 11.5 %, and 47.4 and 64.1 % of field isolates tested, respectively. The anti-sera against the more recently introduced vaccine strains G1 and 431 neutralized 33.3 and 55.1 % (strain G1) or 69.2 and 89.7 % (strain 431) of the field isolates with titres ≥5. [corrected]. Dual vaccine strains displayed a higher cross-neutralization. This study confirms previous observations that more recently introduced vaccine strains induce antibodies with a higher neutralizing capacity compared to vaccine strains that have been used extensively over a long period of time. This study also suggests that dual FCV vaccine strains might neutralize more field isolates compared to single vaccine strains. Vaccine strains should ideally be selected based on updated knowledge on the antigenic properties of field isolates in the local setting, and there is thus a need for continuously studying the evolution of FCV together with the neutralizing capacity of vaccine strain induced antibodies against field isolates at a national and/or regional level.

  8. Biomarker analysis of American toad (Anaxyrus americanus) ...

    EPA Pesticide Factsheets

    The objective of the current study was to use a biomarker-based approach to investigate the influence of atrazine exposure on American toad (Anaxyrus americanus) and grey tree frog (Hyla versicolor) tadpoles. Atrazine is one of the most frequently detected herbicides in environmental matrices throughout the United States. In surface waters, it has been found at concentrations from 0.04–2859 μg/L and thus presents a likely exposure scenario for non-target species such as amphibians. Studies have examined the effect of atrazine on the metamorphic parameters of amphibians, however, the data are often contradictory. Gosner stage 22–24 tadpoles were exposed to 0 (control), 10, 50, 250 or 1250 μg/L of atrazine for 48 h. Endogenous polar metabolites were extracted and analyzed using gas chromatography coupled with mass spectrometry. Statistical analyses of the acquired spectra with machine learning classification models demonstrated identifiable changes in the metabolomic profiles between exposed and control tadpoles. Support vector machine models with recursive feature elimination created a more efficient, non-parametric data analysis and increased interpretability of metabolomic profiles. Biochemical fluxes observed in the exposed groups of both A. americanus and H. versicolor displayed perturbations in a number of classes of biological macromolecules including fatty acids, amino acids, purine nucleosides, pyrimidines, and mono- and di-saccharides. Metabolomic

  9. Biomarker analysis of American toad (Anaxyrus americanus) ...

    EPA Pesticide Factsheets

    The objective of the current study was to use a biomarker-based approach to investigate the influence of atrazine exposure on American toad (Anaxyrus americanus) and grey tree frog (Hyla versicolor) tadpoles. Atrazine is one of the most frequently detected herbicides in environmental matrices throughout the United States. In surface waters, it has been found at concentrations from 0.04–2859 μg/L and thus presents a likely exposure scenario for non-target species such as amphibians. Studies have examined the effect of atrazine on the metamorphic parameters of amphibians, however, the data are often contradictory. Gosner stage 22–24 tadpoles were exposed to 0 (control), 10, 50, 250 or 1250 μg/L of atrazine for 48 h. Endogenous polar metabolites were extracted and analyzed using gas chromatography coupled with mass spectrometry. Statistical analyses of the acquired spectra with machine learning classification models demonstrated identifiable changes in the metabolomic profiles between exposed and control tadpoles. Support vector machine models with recursive feature elimination created a more efficient, non-parametric data analysis and increased interpretability of metabolomic profiles. Biochemical fluxes observed in the exposed groups of both A. americanus and H. versicolor displayed perturbations in a number of classes of biological macromolecules including fatty acids, amino acids, purine nucleosides, pyrimidines, and mono- and di-saccharides. Metabolomic

  10. Neutral current induced {pi}{sup 0} production and neutrino magnetic moment

    SciTech Connect

    Athar, M. Sajjad; Chauhan, S.; Singh, S. K.

    2008-08-01

    We have studied the total cross section, Q{sup 2}, momentum and angular distributions for pions in the {nu}({nu}) induced {pi}{sup 0} production from nucleons. The calculations have been done for the weak production induced by the neutral current in the standard model and the electromagnetic production induced by neutrino magnetic moment. It has been found that with the present experimental limits on the muon neutrino magnetic moment {mu}{sub {nu}{sub {mu}}}, the electromagnetic contribution to the cross section for the {pi}{sup 0} production is small. The neutrino induced neutral current production of {pi}{sup 0}, while giving an alternative method to study the magnetic moment of neutrino {mu}{sub {nu}{sub {mu}}}, does not provide any improvement over the present experimental limit on {mu}{sub {nu}{sub {mu}}} from the observation of this process in future experiments at T2K and NO{nu}A.

  11. Raman study of the pressure-induced neutral-to-ionic transition in tetrathiafulvalene chloranil

    NASA Astrophysics Data System (ADS)

    Hanfland, M.; Brillante, A.; Girlando, A.; Syassen, K.

    1988-07-01

    The pressure-induced phase transition from a neutral regular stack to an ionic dimerized stack (NR-ID) in the tetrathiafulvalene chloranil charge-transfer crystal has been studied by Raman spectroscopy. An accurate estimate of the change of ionicity during the phase transition is obtained from calculations of mode frequencies which are based on the trimer (NR phase) and dimer (ID phase) models of the electron-molecular-vibration interaction. The possibility of the formation of ionic dimers embedded in neutral chains as precursors of the phase change is discussed.

  12. Structural Constraints of Vaccine-Induced Tier-2 Autologous HIV Neutralizing Antibodies Targeting the Receptor-Binding Site.

    PubMed

    Bradley, Todd; Fera, Daniela; Bhiman, Jinal; Eslamizar, Leila; Lu, Xiaozhi; Anasti, Kara; Zhang, Ruijung; Sutherland, Laura L; Scearce, Richard M; Bowman, Cindy M; Stolarchuk, Christina; Lloyd, Krissey E; Parks, Robert; Eaton, Amanda; Foulger, Andrew; Nie, Xiaoyan; Karim, Salim S Abdool; Barnett, Susan; Kelsoe, Garnett; Kepler, Thomas B; Alam, S Munir; Montefiori, David C; Moody, M Anthony; Liao, Hua-Xin; Morris, Lynn; Santra, Sampa; Harrison, Stephen C; Haynes, Barton F

    2016-01-05

    Antibodies that neutralize autologous transmitted/founder (TF) HIV occur in most HIV-infected individuals and can evolve to neutralization breadth. Autologous neutralizing antibodies (nAbs) against neutralization-resistant (Tier-2) viruses are rarely induced by vaccination. Whereas broadly neutralizing antibody (bnAb)-HIV-Envelope structures have been defined, the structures of autologous nAbs have not. Here, we show that immunization with TF mutant Envs gp140 oligomers induced high-titer, V5-dependent plasma neutralization for a Tier-2 autologous TF evolved mutant virus. Structural analysis of autologous nAb DH427 revealed binding to V5, demonstrating the source of narrow nAb specificity and explaining the failure to acquire breadth. Thus, oligomeric TF Envs can elicit autologous nAbs to Tier-2 HIVs, but induction of bnAbs will require targeting of precursors of B cell lineages that can mature to heterologous neutralization.

  13. Two photon absorption laser induced fluorescence measurements of neutral density in a helicon plasma

    SciTech Connect

    Galante, M. E.; Magee, R. M.; Scime, E. E.

    2014-05-15

    We have developed a new diagnostic based on two-photon absorption laser induced fluorescence (TALIF). We use a high intensity (5 MW/cm{sup 2}), narrow bandwidth (0.1 cm{sup −1}) laser to probe the ground state of neutral hydrogen, deuterium and krypton with spatial resolution better than 0.2 cm, a time resolution of 10 ns, and a measurement cadence of 20 Hz. Here, we describe proof-of-principle measurements in a helicon plasma source that demonstrate the TALIF diagnostic is capable of measuring neutral densities spanning four orders of magnitude; comparable to the edge neutral gradients predicted in the DIII-D tokamak pedestal. The measurements are performed in hydrogen and deuterium plasmas and absolute calibration is accomplished through TALIF measurements in neutral krypton. The optical configuration employed is confocal, i.e., both light injection and collection are accomplished with a single lens through a single optical port in the vacuum vessel. The wavelength resolution of the diagnostic is sufficient to separate hydrogen and deuterium spectra and we present measurements from mixed hydrogen and deuterium plasmas that demonstrate isotopic abundance measurements are feasible. Time resolved measurements also allow us to explore the evolution of the neutral hydrogen density and temperature and effects of wall recycling. We find that the atomic neutral density grows rapidly at the initiation of the discharge, reaching the steady-state value within 1 ms. Additionally, we find that neutral hydrogen atoms are born with 0.08 eV temperatures, not 2 eV as is typically assumed.

  14. CXCL10/IP-10 Neutralization Can Ameliorate Lipopolysaccharide-Induced Acute Respiratory Distress Syndrome in Rats

    PubMed Central

    Lang, Shan; Li, Libing; Wang, Xuning; Sun, Junping; Xue, Xinying; Xiao, Yongjiu; Zhang, Mingyue; Ao, Ting; Wang, Jianxin

    2017-01-01

    The role of C-X-C motif chemokine 10 (CXCL10), a pro-inflammatory factor, in the development of acute respiratory distress syndrome (ARDS) remains unclear. In this study, we explored the role of CXCL10 and the effect of CXCL10 neutralization in lipopolysaccharide (LPS)-induced ARDS in rats. The expression of CXCL10 and its receptor chemokine receptor 3(CXCR3) increased after LPS induction. Moreover, neutralization of CXCL10 ameliorated the severity of ARDS by reducing pulmonary edema, inhibiting the release of inflammatory mediators (IFN-γ, IL-6 and ICAM-1) and limiting inflammatory cells (neutrophils, macrophages, CD8+ T cells) influx into the lung, with a reduction in CXCR3 expression in neutrophils and macrophages. Therefore, CXCL10 could be a potential therapeutic target in LPS-induced ARDS. PMID:28046003

  15. Neutral Current {nu} Induced Reactions in Nuclei at Supernova Neutrino Energies

    SciTech Connect

    Chauhan, S.; Athar, M. Sajjad; Singh, S. K.

    2011-11-23

    We calculate cross sections for the neutral current induced neutrino/antineutrino reaction from {sup 208}Pb target and applied it to study Supernova neutrino event rates. The calculations are done in local density approximation taking into account Pauli blocking, Fermi motion effects and renormalization of weak transition strengths in the nuclear medium. The numerical results for the neutrino nucleus total cross sections have been averaged over the various Supernova neutrino/antineutrino fluxes available in literature.

  16. Spontaneously regressing oral papillomas induce systemic antibodies that neutralize canine oral papillomavirus.

    PubMed

    Ghim, S; Newsome, J; Bell, J; Sundberg, J P; Schlegel, R; Jenson, A B

    2000-06-01

    Canine oral papillomavirus (COPV) infection of naive beagle dogs causes oral papillomas, most of which spontaneously regress. Regressor beagles do not develop new oral papillomas because of COPV type-specific, cell-mediated immunity, COPV neutralizing antibodies, or both. Formalin-fixed native and recombinant COPV vaccines that target the systemic immune system induce neutralizing antibodies that prevent development of oral papillomas. This study was designed to determine whether spontaneously regressing mucosal papillomas also targeted the systemic immune system to induce circulating, neutralizing IgG antibodies that protect against infection by COPV. To accomplish this goal, IgG was fractionated from sera collected from weanling beagles and regressor beagles and tested for conferring protection by passive immunization. Serum was tested by ELISA for antibodies against intact virions and then pooled for passive transfer to naive beagles. Preimmune sera were neither reactive by ELISA nor protective by passive transfer. On the other hand, IgG antibodies from regressor beagles were reactive by ELISA and passive transfer conferred protection against COPV challenge. Circulating IgG antibodies induced by spontaneous regression of canine oral papillomas protect beagles against intraoral infection by COPV, a model for mucosotropic HPV. Copyright 2000 Academic Press.

  17. Reproductive Behavior of Echinothrips americanus (Thysanoptera: Thripidae)

    PubMed Central

    Krueger, Stephanie; Jilge, Marcus; Mound, Laurence

    2017-01-01

    Most Thysanoptera possess a haplo-diploid reproductive mode and reproduce via arrhenotoky. Females can mature eggs successively throughout almost their entire life, but in most terebrantian thrips spermiogenesis is complete by adult male eclosion, and testes contain only mature spermatids. In parasitoid wasps this phenomenon of preadult spermiogenesis is described as prospermatogeny. It is unclear if prospermatogeny and this predetermined sperm quantity have implications for mating strategy and fitness. In this study, we give a detailed description of mating behavior of the thripine species Echinothrips americanus, which largely corresponds with the only available data of another species of this family, Frankliniella occidentalis (Thysanoptera: Thripidae). With investigations using light microscopy, we describe for the first time the chronological sequence of internal processes during copulation. The release of male accessory gland material followed subsequently by spermatozoa indicates production of a female-determined type 1 spermatophore. Despite prospermatogeny, males are able to inseminate 10 females with an equal amount of spermatozoa. It is only the quantity of glandular material that decreases with the number of previous copulations. Based on these new findings, the reproductive strategy of this species is discussed.

  18. Homarus Americanus Stomatogastric Nervous System Dissection

    PubMed Central

    Tobin, Anne-Elise; Bierman, Hilary S.

    2009-01-01

    With the goal of understanding how nervous systems produce activity and respond to the environment, neuroscientists turn to model systems that exhibit the activity of interest and are accessible and amenable to experimental methods. The stomatogastric nervous system (STNS) of the American lobster (Homarus americanus; also know was the Atlantic or Maine lobster) has been established as a model system for studying rhythm generating networks and neuromodulation of networks. The STNS consists of 3 anterior ganglia (2 commissural ganglia and an oesophageal ganglion), containing modulatory neurons that project centrally to the stomatogastric ganglion (STG). The STG contains approximately 30 neurons that comprise two central pattern generating networks, the pyloric and gastric networks that underlie feeding behaviors in crustaceans1,2. While it is possible to study this system in vivo3, the STNS continues to produce its rhythmic activity when isolated in vitro. Physical isolation of the STNS in a dish allows for easy access to the somata in the ganglia for intracellular electrophysiological recordings and to the nerves of the STNS for extracellular recordings. Isolating the STNS is a two-part process. The first part, dissecting the stomach from the animal, is described in an accompanying video article4. In this video article, fine dissection techniques are used to isolate the STNS from the stomach. This procedure results in a nervous system preparation that is available for electrophysiological recordings. PMID:19483669

  19. An Indirect and Dynamically Induced Energy Mechanism in a Plasma-Neutral Atmosphere

    NASA Astrophysics Data System (ADS)

    Hsu, Vicki W.

    Plasma-neutral interactions play an integral role in governing the dynamics and energy of an upper atmosphere. In the Earth's ionosphere-thermosphere (I/T) region, these interactions can produce significant structure in neutral temperature, winds, mass density, and composition. External sources of momentum and energy that directly affect these properties of the upper atmosphere have been extensively investigated, but these investigations are not sufficient to fully describe thermosphere phenomenology. Considering the strong plasma-neutral coupling in the I/T region, an internal and indirect energy mechanism, or a feedback exchange between hydrodynamics and thermodynamics, can contribute to the resultant structure. The complex, indirect consequences of internal momentum changes on neutral gas properties have not been thoroughly assessed due to the multivariate nature of the problem and limited observations. Through the use of the National Center for Atmospheric Research Thermosphere-Ionosphere-Electrodynamics General Circulation Model, this thesis seeks to simulate, understand, and quantify how plasma-neutral interactions affect the energy distribution of the upper thermosphere via an indirect, dynamically induced mechanism, and to establish that this mechanism can explain the existence of thermospheric density features. The main conclusions that stem from this dissertation are as follows: (1) Changes in the field-aligned ion drag force alter neutral temperature and mass density by means of a divergent neutral wind field, commanding the formation, local time, and solar cycle variations of the equatorial thermosphere anomaly trough; (2) Ion and viscous drag forces produce balanced motion with sustained, divergent winds that change thermal and mass density structure through adiabatic heating; (3) A boundary that delineates the lower and upper thermosphere is recognized and deemed the "thermopause", and helium is demonstrated to be an effective dynamic tracer for a

  20. Neutral-depletion-induced asymmetric plasma density profile and momentum transport in a helicon thruster

    NASA Astrophysics Data System (ADS)

    Takahashi, Kazunori; Takao, Yoshinori; Chiba, Aiki; Ando, Akira

    2016-09-01

    Axial momentum lost to a lateral wall of a helicon source is directly measured by using a pendulum force balance, where only the lateral wall is attached to the balance immersed in 60-cm-diam and 1.4-m-long vacuum tank (pumping speed of 300-400 L/s). When operating the source with highly ionized krypton and xenon, the strong density decay along the axis is observed inside the source tube, which seems to be due to the neutral depletion. Under such a condition, a non-negligible loss of the axial momentum to the lateral wall is detected. The presently detected loss of the axial momentum indicates the presence of the ions which are axially accelerated by the electric field in the plasma core and then lost to the lateral wall. Furthermore, the helicon thruster immersed in 1-m-diam and 2-m-long vacuum tank (pumping speed of 4000-5000 L/s) is operated at high rf power up to 5 kW in argon, to demonstrate the neutral-depletion-induced axially asymmetric density profile. Combination between the Langmuir probe and the optical diagnosis indicates that the neutral density at the axial center of the source is reduced to 20% of the initial neutral density. This work is partially supported by grant-in-aid for scientific research (16H04084 and 26247096) from the Japan Society for the Promotion of Science.

  1. Antineutrino-induced charge current quasi-elastic neutral hyperon production in ArgoNeuT

    NASA Astrophysics Data System (ADS)

    Farooq, Saima

    This dissertation presents the first topological study of the charge current quasi-elastic (CCQE) neutral hyperon production induced by antineutrinos in the ArgoNeuT detector, a liquid argon time projection chamber (LArTPC) at Fermilab, using 1.20 x 1020 protons-on- target (POT), in the NuMI beam operating in the low energy antineutrino mode. The total cross section for the CCQE neutral hyperon production is reported at the mean production energy of 3.42 GeV. The event yield in data is consistent with the predicted cross section, sigma = 2.7 x 10-40 cm2: sigma(CCQELambda0+Sigma0 ) = 3.7 +/- 1.9(stat.) +/- 1.5 (sys.) x 10 -40 cm2. The study sets a 90% confidence level (C.L.) upper limit on the total cross section of CCQE neutral hyperon production: sigma(CCQE Lambda0+Sigma0) < 7.3 x10-40 cm 2 at 90% C.L.

  2. Low energy electron induced reactions in fluorinated acetamide - probing negative ions and neutral stable counterparts*

    NASA Astrophysics Data System (ADS)

    Kopyra, Janina; König-Lehmann, Constanze; Illenberger, Eugen; Warneke, Jonas; Swiderek, Petra

    2016-06-01

    Electron impact to trifluoroacetamide (CF3CONH2, TFAA) in the energy range 0-12 eV leads to a variety of negative fragment ions which are formed via dissociative electron attachment (DEA). The underlying reactions range from single bond cleavages to remarkably complex reactions that lead to loss of the neutral units HF, H2O and HNCO as deduced from their directly observed ionic counterparts (M - H2O)-, (M - HF)- and (M - HNCO)-. Also formed are the pseudo-halogen ions CN- and OCN-. All these reactions proceed dominantly via a resonance located near 1 eV, i.e., electrons at subexcitation energies trigger reactions involving multiple bond cleavages. The electron induced generation of the neutral molecules HF, H2O and HNCO in condensed TFAA films is probed by temperature controlled thermal desorption spectrometry (TDS) which can be viewed as a complementary techniques to gas-phase experiments in DEA to directly probe the neutral counterparts. Contribution to the Topical Issue "Advances in Positron and Electron Scattering", edited by Paulo Limao-Vieira, Gustavo Garcia, E. Krishnakumar, James Sullivan, Hajime Tanuma and Zoran Petrovic.

  3. Two Photon Absorption Laser Induced Fluorescence for Neutral Hydrogen Profile Measurements

    SciTech Connect

    Scime, Earl E.

    2016-09-23

    The magnitude and spatial dependence of neutral density in magnetic confinement fusion experiments is a key physical parameter, particularly in the plasma edge. Modeling codes require precise measurements of the neutral density to calculate charge-exchange power losses and drag forces on rotating plasmas. However, direct measurements of the neutral density are problematic. In this work, we proposed to construct a laser-based diagnostic capable of providing spatially resolved measurements of the neutral density in the edge of plasma in the DIII-D tokamak. The diagnostic concept is based on two-photon absorption laser induced fluorescence (TALIF). By injecting two beams of 205 nm light (co or counter propagating), ground state hydrogen (or deuterium or tritium) can be excited from the n = 1 level to the n = 3 level at the location where the two beams intersect. Individually, the beams experience no absorption, and therefore have no difficulty penetrating even dense plasmas. After excitation, a fraction of the hydrogen atoms decay from the n = 3 level to the n = 2 level and emit photons at 656 nm (the Hα line). Calculations based on the results of previous TALIF experiments in magnetic fusion devices indicated that a laser pulse energy of approximately 3 mJ delivered in 5 ns would provide sufficient signal-to-noise for detection of the fluorescence. In collaboration with the DIII-D engineering staff and experts in plasma edge diagnostics for DIII-D from Oak Ridge National Laboratory (ORNL), WVU researchers designed a TALIF system capable of providing spatially resolved measurements of neutral deuterium densities in the DIII-D edge plasma. The laser systems were specified, purchased, and assembled at WVU. The TALIF system was tested on a low-power hydrogen discharge at WVU and the plan was to move the instrument to DIII-D for installation in collaboration with ORNL researchers. After budget cuts at DIII-D, the DIII-D facility declined to support

  4. Measurement of longitudinal emittance growth using a laser-induced neutralization method

    SciTech Connect

    Yuan, V.W.; Garcia, R.; Johnson, K.F.; Saadatmand, K.; Sander, O.R.; Sandoval, D.; Shinas, M.

    1991-01-01

    A laser-induced neutralization technique, LINDA, has been used to study the longitudinal emittance of the 5-MeV H{sup {minus}} beam exiting the drift-tube Linac (DTL) of the Los Alamos Accelerator Test Stand (ATS). By using multiple laser intersection points, longitudinal emittance growths over drift distances of 23.6 and 30.6 cm were measured. Subsequently, a beam transport line, which consisted of one arm of a beam funnel, was substituted for the drift space. Measurements show that the elements of the funnel constrain emittance growth while the H{sup {minus}} beam is contained within these transport elements.

  5. Model of photo-induced neutral-ionic phase transition in organic charge-transfer salts

    NASA Astrophysics Data System (ADS)

    Yartsev, V. M.

    1998-01-01

    One-dimensional donor-acceptor mixed chains are modeled by a periodic DADA tetramer. Electron coupling to intramolecular vibrations are taken into account explicitly. Generalized adiabatic potentials are calculated for the cases of regular and dimerized stacks which are characteristic, respectively, of quasi-neutral (N) and quasi-ionic (I) phases of a tetrathiafulvalene-chloranil compound. A sharp difference in life-times of photo-induced I-states in the N-phase and N-states in the I-phase is discussed within the periodic DADA tetramer model.

  6. The relationship between Pseudodiplorchis americanus (Monogenea) density and host resources under controlled environmental conditions.

    PubMed

    Tocque, K; Tinsley, R C

    1994-02-01

    A previous study has shown that, under natural conditions, energy reserves of the desert toad, Scaphiopus couchii, are negatively related to the density of infection by Pseudodiplorchis americanus. However, this was based predominantly on collections of active animals from breeding congregations and inevitably selected toads which were in good condition. The parasite, a blood-feeding monogenean, occurs in burdens of up to 30 worms/host (mean intensity 6 worms/host) and represents a significant drain on reserves because the host does not feed during a 10-month hibernation. Field studies cannot resolve the possibility that larger worm densities are not observed in nature due to parasite-induced host mortality. The present study was conducted during investigations of P. americanus development and survival under controlled laboratory conditions, utilizing experimentally infected hosts which created worm densities larger than those observed in natural populations. At all temperature regimes, infected animals had smaller fat bodies than those uninfected but differences were generally not statistically significant due to large individual variations, presumably resulting from variations in past feeding efficiency. At cool temperatures (15-20 degrees C) there was no density-dependent effect on host fat body weight, and at a diurnal temperature cycle of 20-34 degrees C (simulating that experienced by host and parasite during the summer months), the effects of high temperatures were greater than the effects of infection, due to increased toad metabolic rates. The most significant effects of P. americanus were observed in hosts that began hibernation in relatively poor condition and experienced moderate temperatures (25 degrees C) during hibernation. The toads generally maintained packed blood cell levels (PCV) levels even when fat body weights were low, but infected animals had a lower PCV irrespective of fab body levels. In animals unfed after field collection, PCV was

  7. P53-dependent upregulation of neutral sphingomyelinase-2: role in doxorubicin-induced growth arrest

    PubMed Central

    Shamseddine, A A; Clarke, C J; Carroll, B; Airola, M V; Mohammed, S; Rella, A; Obeid, L M; Hannun, Y A

    2015-01-01

    Neutral sphingomyelinase-2 (nSMase2) is a ceramide-generating enzyme that has been implicated in growth arrest, apoptosis and exosome secretion. Although previous studies have reported transcriptional upregulation of nSMase2 in response to daunorubicin, through Sp1 and Sp3 transcription factors, the role of the DNA damage pathway in regulating nSMase2 remains unclear. In this study, we show that doxorubicin induces a dose-dependent induction of nSMase2 mRNA and protein with concomitant increases in nSMase activity and ceramide levels. Upregulation of nSMase2 was dependent on ATR, Chk1 and p53, thus placing it downstream of the DNA damage pathway. Moreover, overexpression of p53 was sufficient to transcriptionally induce nSMase2, without the need for DNA damage. DNA-binding mutants as well as acetylation mutants of p53 were unable to induce nSMase2, suggesting a role of nSMase2 in growth arrest. Moreover, knockdown of nSMase2 prevented doxorubicin-induced growth arrest. Finally, p53-induced nSMase2 upregulation appears to occur via a novel transcription start site upstream of exon 3. These results identify nSMase2 as a novel p53 target gene, regulated by the DNA damage pathway to induce cell growth arrest. PMID:26512957

  8. Trichinella surveillance in black bears (Ursus americanus) from Oregon, USA.

    PubMed

    Mortenson, J A; Kent, M L; Fowler, D R; Chomel, B B; Immell, D A

    2014-01-01

    We used serology and muscle digestion to test black bears (Ursus americanus) from western Oregon, USA, for Trichinella. Results indicate black bears in Oregon are not part of a sylvatic cycle for Trichinella, and risk of human exposure to Trichinella larvae from eating black bear meat from Oregon appears low.

  9. Sarcocystis oreamni n. sp. from the mountain goat (oreamnos americanus)

    USDA-ARS?s Scientific Manuscript database

    Numerous species of Sarcocystis have been reported from wild ruminants but none has been named from the Rocky Mountain goat (Oreamnos americanus). Mature sarcocysts were found in frozen muscle samples of 3 of 7 mountain goats from Alaska, USA. Two morphological types of sarcocysts were found; 1 had ...

  10. The Anti-Fibrotic Effect of Mycophenolic Acid–Induced Neutral Endopeptidase

    PubMed Central

    Dell'Oglio, Maria Pia; Rossini, Michele; Divella, Chiara; Pontrelli, Paola; Verrienti, Raffaella; Rutigliano, Monica; Ditonno, Pasquale; Stifanelli, Patrizia; Ancona, Nicola; Schena, Francesco Paolo; Grandaliano, Giuseppe

    2010-01-01

    Mycophenolic acid (MPA) appears to have anti-fibrotic effects, but the molecular mechanisms underlying this are unknown. We prospectively studied 35 stable kidney transplant recipients maintained on cyclosporine and azathioprine. We converted 20 patients from azathioprine to enteric-coated mycophenolate sodium (EC-MPS) and continued the remaining 15 patients on azathioprine. Exploratory mRNA expression profiling, performed on five randomly selected EC-MPS patients, revealed significant upregulation of neutral endopeptidase (NEP), which is an enzyme that degrades angiotensin II. We confirmed these microarray data by measuring levels of NEP expression in all subjects; in addition, we found that NEP gene expression correlated inversely with proteinuria. In an additional 33 patients, glomerular and tubular NEP protein levels from renal graft biopsies were significantly higher among the 13 patients receiving cyclosporine + EC-MPS than among the 12 patients receiving cyclosporine + azathioprine or 8 patients receiving cyclosporine alone. Glomerular NEP expression inversely correlated with glomerulosclerosis and proteinuria, and tubular NEP expression inversely correlated with interstitial fibrosis. Incubation of human proximal tubular cells with MPA increased NEP gene expression in a dose- and time-dependent manner. Moreover, MPA reduced angiotensin II–induced expression of the profibrotic factor plasminogen activator inhibitor-1, and a specific NEP inhibitor completely reversed this effect. Taken together, our data suggest that MPA directly induces expression of neutral endopeptidase, which may reduce proteinuria and slow the progression of renal damage in kidney transplant recipients. PMID:20864690

  11. Apoptosis induced by islet amyloid polypeptide soluble oligomers is neutralized by diabetes-associated specific antibodies

    PubMed Central

    Bram, Yaron; Frydman-Marom, Anat; Yanai, Inbal; Gilead, Sharon; Shaltiel-Karyo, Ronit; Amdursky, Nadav; Gazit, Ehud

    2014-01-01

    Soluble oligomeric assemblies of amyloidal proteins appear to act as major pathological agents in several degenerative disorders. Isolation and characterization of these oligomers is a pivotal step towards determination of their pathological relevance. Here we describe the isolation of Type 2 diabetes-associated islet amyloid polypeptide soluble cytotoxic oligomers; these oligomers induced apoptosis in cultured pancreatic cells, permeated model lipid vesicles and interacted with cell membranes following complete internalization. Moreover, antibodies which specifically recognized these assemblies, but not monomers or amyloid fibrils, were exclusively identified in diabetic patients and were shown to neutralize the apoptotic effect induced by these oligomers. Our findings support the notion that human IAPP peptide can form highly toxic oligomers. The presence of antibodies identified in the serum of diabetic patients confirms the pathological relevance of the oligomers. In addition, the newly identified structural epitopes may also provide new mechanistic insights and a molecular target for future therapy. PMID:24589570

  12. Apoptosis induced by islet amyloid polypeptide soluble oligomers is neutralized by diabetes-associated specific antibodies.

    PubMed

    Bram, Yaron; Frydman-Marom, Anat; Yanai, Inbal; Gilead, Sharon; Shaltiel-Karyo, Ronit; Amdursky, Nadav; Gazit, Ehud

    2014-03-04

    Soluble oligomeric assemblies of amyloidal proteins appear to act as major pathological agents in several degenerative disorders. Isolation and characterization of these oligomers is a pivotal step towards determination of their pathological relevance. Here we describe the isolation of Type 2 diabetes-associated islet amyloid polypeptide soluble cytotoxic oligomers; these oligomers induced apoptosis in cultured pancreatic cells, permeated model lipid vesicles and interacted with cell membranes following complete internalization. Moreover, antibodies which specifically recognized these assemblies, but not monomers or amyloid fibrils, were exclusively identified in diabetic patients and were shown to neutralize the apoptotic effect induced by these oligomers. Our findings support the notion that human IAPP peptide can form highly toxic oligomers. The presence of antibodies identified in the serum of diabetic patients confirms the pathological relevance of the oligomers. In addition, the newly identified structural epitopes may also provide new mechanistic insights and a molecular target for future therapy.

  13. Neutralization of Tumor Necrosis Factor Bioactivity Ameliorates Urethane-Induced Pulmonary Oncogenesis in Mice1

    PubMed Central

    Karabela, Sophia P; Kairi, Chrysoula A; Magkouta, Sophia; Psallidas, Ioannis; Moschos, Charalampos; Stathopoulos, Ioannis; Zakynthinos, Spyros G; Roussos, Charis; Kalomenidis, Ioannis; Stathopoulos, Georgios T

    2011-01-01

    Tumor necrosis factor (TNF) has been implicated in inflammation-associated tumor progression. Although multiple reports identified a role for TNF signaling in established cancers, few studies have assessed the impact of TNF blockade on early tumor formation promotion. We aimed at exploring the effects of TNF neutralization in a preclinical mouse model of lung carcinogenesis. For this, Balb/c mice (n = 42) received four weekly intraperitoneal urethane injections (1 g/kg) and twice-weekly intraperitoneal soluble TNF receptor (etanercept; 10 mg/kg) administered during tumor initiation/promotion, tumor progression, or continuously (months 1, 6, and 1–8 after urethane start, respectively). Lung oncogenesis was assessed after 8 months. In separate short-term studies, Balb/c mice (n = 21) received a single control or urethane injection followed by twice-weekly intraperitoneal control or sTNFR:Fc injections. Lung inflammation was assessed after 1 week. We found that sTNFR:Fc treatment during tumor initiation/promotion resulted in a significant reduction of tumor number but not dimensions. However, sTNFR:Fc administered during tumor progression did not impact tumor multiplicity but significantly decreased tumor diameter. Continued sTNFR:Fc administration was effective in halting both respiratory tumor formation and progression in response to urethane. This favorable impact was associated with impaired cellular proliferation and new vessel formation in lung tumors. In addition, TNF neutralization altered the lung inflammatory response to urethane, evidenced by reductions in TNF and macrophage and increases in interferon γ and interleukin 10 content of the air spaces. sTNFR:Fc treatment of RAW264.7 macrophages downregulated TNF and enhanced interferon γ and interleukin 10 expression. In conclusion, TNF neutralization is effective against urethane-induced lung oncogenesis in mice and could present a lung chemoprevention strategy worth testing clinically. PMID:22241960

  14. Extreme ultraviolet emission from laser-induced plasma relevance to neutral gas environment simulation in LEO

    NASA Astrophysics Data System (ADS)

    Tagawa, Masahito; Kimoto, Yugo; Yokota, Kumiko; Ohira, Junki; Watanabe, Daiki; Nishimura, Hiroaki

    The reaction mechanism of atomic oxygen (AO) in low Earth orbit (LEO) with spacecraft materials has been studied by ground-based experiments using laser-detonation hyperthermal beam source, which enables to accelerate the electrically neutral AO up to 8 km/s. However, the beam conditions in the laser-detonation sources could not fully duplicate the AO environment in space. The difference in beam condition including side products leads to the different material responses. The light emission from the laser-induced oxygen plasma may affect the erosion of ultraviolet (UV)-sensitive materials. However, the light emission could also be used as a diagnostic tool to understand the molecular processes in plasma. In this presentation, extreme ultraviolet (EUV) emission from the laser-induced plasma during AO test was evaluated by the flat field EUV spectrometer. Many emission lines between 25-40 nm originated from OII and OIII were observed from the laser-induced oxygen plasma. This result suggested multiple-charged O ions are generated in the laser-induced plasma. Promotion of oxygen dissociation effect by adding Ar in the target gas was explained by the energy transfer processes from Ar to O2 in the plasma. From the viewpoint of reducing the side products in the AO exposure tests, a method to reduce the EUV emission will also be investigated. These results could be used for establishing more accurate ground-based natural gas simulations on the space environmental effect of materials.

  15. AAV natural infection induces broad cross-neutralizing antibody responses to multiple AAV serotypes in chimpanzees.

    PubMed

    Calcedo, Roberto; Wilson, James M

    2016-06-01

    Cross-sectional studies of primates have revealed that natural neutralizing antibody (NAb) responses to adeno-associated viruses (AAV) span multiple serotypes. This differs from the phenotype of the NAb response to an AAV vector delivered to sero-negative nonhuman primates which is typically restricted to the administered AAV serotype. To better understand the mechanism by which natural AAV infections result in broad NAb responses, we conducted a longitudinal study spanning 10 years in which we evaluated serum-circulating AAV NAb levels in captive-housed chimpanzees. In a cohort of 25 chimpanzees we identified three distinct groups of animals: those which never sero-converted to AAV (naïve); those which were persistently seropositive (chronic); and those that seroconverted during the 10 year period (acute). For the chronic group we found a broad sero-response characterized by NAbs reacting to multiple AAV serotypes. A similar cross-neutralization pattern of NAbs was observed in the acute group. These data support our hypothesis that a single natural infection with AAV induces a broadly cross-reactive NAb response to multiple AAV serotypes.

  16. AAV Natural Infection Induces Broad Cross-Neutralizing Antibody Responses to Multiple AAV Serotypes in Chimpanzees.

    PubMed

    Calcedo, Roberto; Wilson, James M

    2016-06-01

    Cross-sectional studies of primates have revealed that natural neutralizing antibody (NAb) responses to adeno-associated viruses (AAV) span multiple serotypes. This differs from the phenotype of the NAb response to an AAV vector delivered to seronegative nonhuman primates that is typically restricted to the administered AAV serotype. To better understand the mechanism by which natural AAV infections result in broad NAb responses, we conducted a longitudinal study spanning 10 years in which we evaluated serum-circulating AAV NAb levels in captive-housed chimpanzees. In a cohort of 25 chimpanzees we identified 3 distinct groups of animals: those that never seroconverted to AAV (naïve), those that were persistently seropositive (chronic), and those that seroconverted during the 10-year period (acute). For the chronic group we found a broad seroresponse characterized by NAbs reacting to multiple AAV serotypes. A similar cross-neutralization pattern of NAbs was observed in the acute group. These data support our hypothesis that a single natural infection with AAV induces a broadly cross-reactive NAb response to multiple AAV serotypes.

  17. Laser Induced Fluorescence Spectroscopy of Neutral and Ionized Polycyclic Aromatic Hydrocarbons in the Cosmic Simulation Chamber

    NASA Technical Reports Server (NTRS)

    Bejaoui, Salma; Salama, Farid; Contreras, Cesar; Sciamma O'Brien, Ella; Foing, Bernard; Pascale, Ehrenfreund

    2015-01-01

    Polycyclic aromatic hydrocarbon (PAH) molecules are considered the best carriers to account for the ubiquitous infrared emission bands. PAHs have also been proposed as candidates to explain the diffuse interstellar bands (DIBs), a series of absorption features seen on the interstellar extinction curve and are plausible carriers for the extended red emission (ERE), a photoluminescent process associated with a wide variety of interstellar environments. Extensive efforts have been devoted over the past two decades to characterize the physical and chemical properties of PAH molecules and ions in space. Absorption spectra of PAH molecules and ions trapped in solid matrices have been compared to the DIBs. Absorption spectra of several cold, isolated gas-phase PAHs have also been measured under experimental conditions that mimic the interstellar conditions. The purpose of this study is to provide a new dimension to the existing spectroscopic database of neutral and single ionized PAHs that is largely based on absorption spectra by adding emission spectroscopy data. The measurements are based on the laser induced fluorescence (LIF) technique and are performed with the Pulsed Discharge Nozzle (PDN) of the COSmIC laboratory facility at NASA Ames laboratory. The PDN generates a plasma in a free supersonic jet expansion to simulate the physical and the chemical conditions in interstellar environments. We focus, here, on the fluorescence spectra of large neutral PAHs and their cations where there is a lack of fluorescence spectroscopy data. The astronomical implications of the data (e.g., ERE) are examined.

  18. Comparative tissue expression of American lobster (Homarus americanus) immune genes during bacterial and scuticociliate challenge.

    PubMed

    Clark, K Fraser; Acorn, Adam R; Wang, Haili; Greenwood, Spencer J

    2015-12-01

    The American lobster (Homarus americanus) fishery is the most economically significant fishery in Canada; although comparatively little is known about the lobsters' response to pathogenic challenge. This is the first study to investigate the expression of immune genes in tissues outside of the lobster hepatopancreas in response to challenges by the Gram-positive bacteria, Aerococcus viridans var. homari or the scuticociliate parasite, Anophryoides haemophila. The hepatopancreas has been regarded as the major humoral immune organ in crustaceans, but the contribution of other organs and tissues to the molecular immune response has largely been overlooked. This study used RT-qPCR to monitor the gene expression of several immune genes including three anti-lipopolysaccharide isoforms (ALF) Homame ALF-B1, Homame ALF-C1 and ALFHa-1, acute phase serum amyloid protein A (SAA), as well as thioredoxin and hexokinase, in antennal gland and gill tissues. Our findings indicate that the gene expression of the SAA and all ALF isoforms in the antennal gland and gill tissues increased in response to pathogenic challenge. However, there was differential expression of individual ALF isoforms that were dependent on both the tissue, and the pathogen used in the challenge. The gene expression changes of several immune genes were found to be higher in the antennal gland than have been previously reported for the hepatopancreas. This study demonstrates that increased immune gene expression from the gill and antennal gland over the course of pathogen induced disease contributes to the immune response of H. americanus.

  19. Mild pressure induces rapid accumulation of neutral lipid (triacylglycerol) in Chlorella spp.

    PubMed

    Praveenkumar, Ramasamy; Kim, Bohwa; Lee, Jiye; Vijayan, Durairaj; Lee, Kyubock; Nam, Bora; Jeon, Sang Goo; Kim, Dong-Myung; Oh, You-Kwan

    2016-11-01

    Effective enhancement of neutral lipid (especially triacylglycerol, TAG) content in microalgae is an important issue for commercialization of microalgal biorefineries. Pressure is a key physical factor affecting the morphological, physiological, and biochemical behaviors of organisms. In this paper, we report a new stress-based method for induction of TAG accumulation in microalgae (specifically, Chlorella sp. KR-1 and Ch. sp. AG20150) by very-short-duration application of mild pressure. Pressure treatments of 10-15bar for 2h resulted in a considerable, ∼55% improvement of the 10-100g/Lcells' TAG contents compared with the untreated control. The post-pressure-treatment increase of cytoplasmic TAG granules was further confirmed by transmission electron microscopy (TEM). Notwithstanding the increased TAG content, the total lipid content was not changed by pressurization, implying that pressure stress possibly induces rapid remodeling/transformation of algal lipids rather than de novo biosynthesis of TAG.

  20. Disorder-induced heating of ultracold neutral plasmas created from atoms in partially filled optical lattices

    NASA Astrophysics Data System (ADS)

    Murphy, D.; Sparkes, B. M.

    2016-08-01

    We quantify the disorder-induced heating (DIH) of ultracold neutral plasmas (UCNPs) created from cold atoms in optical lattices with partial filling fractions, using a conservation of energy model involving the spatial correlations of the initial state and the equation of state in thermal equilibrium for a one-component plasma. We show, for experimentally achievable filling fractions, that the ionic Coulomb coupling parameter could be increased to a degree comparable to other proposed DIH-mitigation schemes. Molecular dynamics simulations were performed with compensation for finite-size and periodic boundary effects, which agree with calculations using the model. Reduction of DIH using optical lattices will allow for the study of strongly coupled plasma physics using low-density, low-temperature, laboratory-based plasmas, and lead to improved brightness in UCNP-based cold electron and ion beams, where DIH is otherwise a fundamental limitation to beam focal sizes and diffraction imaging capability.

  1. Alkaline, acid, and neutral phosphatase activities are induced during development in Myxococcus xanthus.

    PubMed

    Weinberg, R A; Zusman, D R

    1990-05-01

    One of the signals that has been reported to be important in stimulating fruiting body formation of Myxococcus xanthus is starvation for phosphate. We therefore chose to study phosphatase activity during M. xanthus development. Many phosphatases can cleave the substrate p-nitrophenol phosphate. Using this substrate in buffers at various pHs, we obtained a profile of phosphatase activities during development and germination of M. xanthus. These experiments indicated that there are five patterns of phosphatase activity in M. xanthus: two vegetative and three developmental. The two uniquely vegetative activities have pH optima at 7.2 and 8.5. Both require magnesium and both are inhibited by the reducing agent dithiothreitol. The developmental (spores) patterns of activity have pH optima of 5.2, 7.2, and 8.5. All three activities are Mg independent. Only the alkaline phosphatase activity is inhibited by dithiothreitol. The acid phosphatase activity is induced very early in development, within the first 2 to 4 h. Both the neutral and alkaline phosphatase Mg-independent activities are induced much later, about the time that myxospores become evident (24 to 30 h). The three activities are greatly diminished upon germination; however, the kinetics of loss differ for all three. The acid phosphatase activity declines very rapidly, the neutral activity begins to decline only after spores begin to convert to rods, and the alkaline phosphatase activity remains high until the time the cells begin to divide. All three developmental activities were measured in the developmental signalling mutants carrying asg, csg, and dsg. The pattern of expression obtained in the mutants was consistent with that of other developmentally regulated genes which exhibit similar patterns of expression during development. The ease with which phosphatases can be assayed should make the activities described in this report useful biochemical markers of stages of both fruiting body formation and

  2. The role of neutral endopeptidase in caerulein-induced acute pancreatitis.

    PubMed

    Koh, Yung-Hua; Moochhala, Shabbir; Bhatia, Madhav

    2011-11-15

    Substance P (SP) is well known to promote inflammation in acute pancreatitis (AP) by interacting with neurokinin-1 receptor. However, mechanisms that terminate SP-mediated responses are unclear. Neutral endopeptidase (NEP) is a cell-surface enzyme that degrades SP in the extracellular fluid. In this study, we examined the expression and the role of NEP in caerulein-induced AP. Male BALB/c mice (20-25 g) subjected to 3-10 hourly injections of caerulein (50 μg/kg) exhibited reduced NEP activity and protein expression in the pancreas and lungs. Additionally, caerulein (10(-7) M) also downregulated NEP activity and mRNA expression in isolated pancreatic acinar cells. The role of NEP in AP was examined in two opposite ways: inhibition of NEP (phosphoramidon [5 mg/kg] or thiorphan [10 mg/kg]) followed by 6 hourly caerulein injections) or supplementation with exogenous NEP (10 hourly caerulein injections, treatment of recombinant mouse NEP [1 mg/kg] during second caerulein injection). Inhibition of NEP raised SP levels and exacerbated inflammatory conditions in mice. Meanwhile, the severity of AP, determined by histological examination, tissue water content, myeloperoxidase activity, and plasma amylase activity, was markedly better in mice that received exogenous NEP treatment. Our results suggest that NEP is anti-inflammatory in caerulein-induced AP. Acute inhibition of NEP contributes to increased SP levels in caerulein-induced AP, which leads to augmented inflammatory responses in the pancreas and associated lung injury.

  3. Recombinant influenza H7 hemagglutinins induce lower neutralizing antibody titers in mice than do seasonal hemagglutinins

    PubMed Central

    Blanchfield, Kristy; Kamal, Ram P; Tzeng, Wen-Pin; Music, Nedzad; Wilson, Jason R; Stevens, James; Lipatov, Aleksander S; Katz, Jacqueline M; York, Ian A

    2014-01-01

    Background Vaccines against avian influenza viruses often require high hemagglutinin (HA) doses or adjuvants to achieve serological titers associated with protection against disease. In particular, viruses of the H7 subtype frequently do not induce strong antibody responses following immunization. Objectives To evaluate whether poor immunogenicity of H7 viruses is an intrinsic property of the H7 hemagglutinin. Methods We compared the immunogenicity, in naïve mice, of purified recombinant HA from two H7 viruses [A/Netherlands/219/2003(H7N7) and A/New York/107/2003(H7N2)] to that of HA from human pandemic [A/California/07/2009(H1N1pdm09)] and seasonal [A/Perth16/2009(H3N2)] viruses. Results After two intramuscular injections with purified hemagglutinin, mice produced antibodies to all HAs, but the response to the human virus HAs was greater than to H7 HAs. The difference was relatively minor when measured by ELISA, greater when measured by hemagglutination inhibition assays, and more marked still by microneutralization assays. H7 HAs induced little or no neutralizing antibody response in mice at either dose tested. Antibodies induced by H7 were of significantly lower avidity than for H3 or H1N1pdm09. Conclusions We conclude that H7 HAs may be intrinsically less immunogenic than HA from seasonal human influenza viruses. PMID:25213778

  4. HPV16/18 L1 VLP Vaccine Induces Cross-Neutralizing Antibodies that May Mediate Cross-Protection

    PubMed Central

    Kemp, Troy J.; Hildesheim, Allan; Safaeian, Mahboobeh; Dauner, Joseph G.; Pan, Yuanji; Porras, Carolina; Schiller, John T.; Lowy, Douglas R.; Herrero, Rolando; Pinto, Ligia A.

    2011-01-01

    Human papillomavirus (HPV) L1 VLP-based vaccines are protective against HPV vaccine-related types; however, the correlates of protection have not been defined. We observed that vaccination with Cervarix™ induced cross-neutralizing antibodies for HPV types for which evidence of vaccine efficacy has been demonstrated (HPV31/45) but not for other types (HPV52/58). In addition, HPV31/45 cross-neutralizing titers showed a significant increase with number of doses (HPV31, p<0.001; HPV45, p<0.001) and correlated with HPV16/18 neutralizing titers, respectively. These findings raise the possibility that cross-neutralizing antibodies are effectors of cross-protection observed for the HPV16/18 vaccine. PMID:21241731

  5. Safety of florfenicol in the adult lobster (Homarus americanus).

    PubMed

    Basti, David; Bouchard, Deborah; Lichtenwalner, Anne

    2011-03-01

    Aerococcus viridans, the causative agent of the disease gaffkemia, was a major cause of mortality in lobsters (Homarus americanus) held in tidal impoundments during the 1970s and 1980s. Despite reports of an increase in the mortality of lobsters during impoundment, and the widespread prophylactic use of oxytetracycline against A. viridans, this bacterium has not been detected in active disease surveillance of the Maine postcapture lobster population. However, Photobacterium indicum may be an emerging opportunistic pathogen of stressed lobsters. An acute toxicity trial was conducted as a rapid screening procedure for the potential future use of the antibiotic florfenicol. Based on the results of this experiment, florfenicol appears to be well tolerated in adult H. americanus by intrapericardial injection at the 10-100 mg/kg dose. Oxytetracycline dihydrate is contraindicated by intrapericardial injection at the 10-100 mg/kg dose.

  6. Estimating Collisionally-Induced Escape Rates of Light Neutrals from Early Mars

    NASA Astrophysics Data System (ADS)

    Gacesa, M.; Zahnle, K. J.

    2016-12-01

    Collisions of atmospheric gases with hot oxygen atoms constitute an important non-thermal mechanism of escape of light atomic and molecular species at Mars. In this study, we present revised theoretical estimates of non-thermal escape rates of neutral O, H, He, and H2 based on recent atmospheric density profiles obtained from the NASA Mars Atmosphere and Volatile Evolution (MAVEN) mission and related theoretical models. As primary sources of hot oxygen, we consider dissociative recombination of O2+ and CO2+ molecular ions. We also consider hot oxygen atoms energized in primary and secondary collisions with energetic neutral atoms (ENAs) produced in charge-exchange of solar wind H+ and He+ ions with atmospheric gases1,2. Scattering of hot oxygen and atmospheric species of interest is modeled using fully-quantum reactive scattering formalism3. This approach allows us to construct distributions of vibrationally and rotationally excited states and predict the products' emission spectra. In addition, we estimate formation rates of excited, translationally hot hydroxyl molecules in the upper atmosphere of Mars. The escape rates are calculated from the kinetic energy distributions of the reaction products using an enhanced 1D model of the atmosphere for a range of orbital and solar parameters. Finally, by considering different scenarios, we estimate the influence of these escape mechanisms on the evolution of Mars's atmosphere throughout previous epochs and their impact on the atmospheric D/H ratio. M.G.'s research was supported by an appointment to the NASA Postdoctoral Program at the NASA Ames Research Center, administered by Universities Space Research Association under contract with NASA. 1N. Lewkow and V. Kharchenko, "Precipitation of Energetic Neutral Atoms and Escape Fluxes induced from the Mars Atmosphere", Astroph. J., 790, 98 (2014) 2M. Gacesa, N. Lewkow, and V. Kharchenko, "Non-thermal production and escape of OH from the upper atmosphere of Mars", arXiv:1607

  7. Precipitation of energetic neutral atoms and induced non-thermal escape fluxes from the Martian atmosphere

    SciTech Connect

    Lewkow, N. R.; Kharchenko, V.

    2014-08-01

    The precipitation of energetic neutral atoms, produced through charge exchange collisions between solar wind ions and thermal atmospheric gases, is investigated for the Martian atmosphere. Connections between parameters of precipitating fast ions and resulting escape fluxes, altitude-dependent energy distributions of fast atoms and their coefficients of reflection from the Mars atmosphere, are established using accurate cross sections in Monte Carlo (MC) simulations. Distributions of secondary hot (SH) atoms and molecules, induced by precipitating particles, have been obtained and applied for computations of the non-thermal escape fluxes. A new collisional database on accurate energy-angular-dependent cross sections, required for description of the energy-momentum transfer in collisions of precipitating particles and production of non-thermal atmospheric atoms and molecules, is reported with analytic fitting equations. Three-dimensional MC simulations with accurate energy-angular-dependent cross sections have been carried out to track large ensembles of energetic atoms in a time-dependent manner as they propagate into the Martian atmosphere and transfer their energy to the ambient atoms and molecules. Results of the MC simulations on the energy-deposition altitude profiles, reflection coefficients, and time-dependent atmospheric heating, obtained for the isotropic hard sphere and anisotropic quantum cross sections, are compared. Atmospheric heating rates, thermalization depths, altitude profiles of production rates, energy distributions of SH atoms and molecules, and induced escape fluxes have been determined.

  8. Somatic Hypermutation-Induced Changes in the Structure and Dynamics of HIV-1 Broadly Neutralizing Antibodies.

    PubMed

    Davenport, Thaddeus M; Gorman, Jason; Joyce, M Gordon; Zhou, Tongqing; Soto, Cinque; Guttman, Miklos; Moquin, Stephanie; Yang, Yongping; Zhang, Baoshan; Doria-Rose, Nicole A; Hu, Shiu-Lok; Mascola, John R; Kwong, Peter D; Lee, Kelly K

    2016-07-20

    Antibody somatic hypermutation (SHM) and affinity maturation enhance antigen recognition by modifying antibody paratope structure to improve its complementarity with the target epitope. SHM-induced changes in paratope dynamics may also contribute to antibody maturation, but direct evidence of this is limited. Here, we examine two classes of HIV-1 broadly neutralizing antibodies (bNAbs) for SHM-induced changes in structure and dynamics, and delineate the effects of these changes on interactions with the HIV-1 envelope glycoprotein (Env). In combination with new and existing structures of unmutated and affinity matured antibody Fab fragments, we used hydrogen/deuterium exchange with mass spectrometry to directly measure Fab structural dynamics. Changes in antibody structure and dynamics were positioned to improve complementarity with Env, with changes in dynamics primarily observed at the paratope peripheries. We conclude that SHM optimizes paratope complementarity to conserved HIV-1 epitopes and restricts the mobility of paratope-peripheral residues to minimize clashes with variable features on HIV-1 Env.

  9. Viscum album neutralizes tumor-induced immunosuppression in a human in vitro cell model

    PubMed Central

    Steinborn, Carmen; Klemd, Amy Marisa; Sauer, Barbara; Garcia-Käufer, Manuel; Urech, Konrad; Follo, Marie; Ücker, Annekathrin; Kienle, Gunver Sophia; Huber, Roman

    2017-01-01

    Tumor cells have the capacity to secrete immunosuppressive substances in order to diminish dendritic cell (DC) activity and thereby escape from immune responses. The impact of mistletoe (Viscum album) extracts (VAE), which are frequently used as an additive anti-cancer therapy to stimulate the immune response, is still unknown. Using a human cellular system, the impact of two different VAE (VAEA + VAEI) on the maturation of human dendritic cells and on T cell function has been investigated using flow cytometry, automated fluorescence microscopy and cytokine bead array assays. Furthermore, we examined whether VAEI was able to counteract tumor-induced immunosuppression within this cellular system using a renal cancer cell model. The role of mistletoe lectin (ML) was analyzed using ML-specific antibodies and ML-depleted VAEI. VAEI and VAEA augmented the maturation of dendritic cells. VAEI abrogated tumor-induced immunosuppression of dendritic cells and both processes were partially mediated by ML since ML-depleted VAEI and ML-specific antibodies almost neutralized the rehabilitative effects of VAEI on DC maturation. Using these settings, co-culture experiments with purified CD4+ T cells had no influence on T cell proliferation and activation but did have an impact on IFN-γ secretion. The study provides a potential mode-of-action of VAE as an additive cancer therapy based on immunomodulatory effects. However, the impact on the in vivo situation has to be evaluated in further studies. PMID:28719632

  10. Inactivated H7 Influenza Virus Vaccines Protect Mice despite Inducing Only Low Levels of Neutralizing Antibodies.

    PubMed

    Kamal, Ram P; Blanchfield, Kristy; Belser, Jessica A; Music, Nedzad; Tzeng, Wen-Pin; Holiday, Crystal; Burroughs, Ashley; Sun, Xiangjie; Maines, Taronna R; Levine, Min Z; York, Ian A

    2017-10-15

    Avian influenza viruses of the H7 hemagglutinin (HA) subtype present a significant public health threat, as evidenced by the ongoing outbreak of human A(H7N9) infections in China. When evaluated by hemagglutination inhibition (HI) and microneutralization (MN) assays, H7 viruses and vaccines are found to induce lower level of neutralizing antibodies (nAb) than do their seasonal counterparts, making it difficult to develop and evaluate prepandemic vaccines. We have previously shown that purified recombinant H7 HA appear to be poorly immunogenic in that they induce low levels of HI and MN antibodies. In this study, we immunized mice with whole inactivated reverse genetics reassortant (RG) viruses expressing HA and neuraminidase (NA) from 3 different H7 viruses [A/Shanghai/2/2013(H7N9), A/Netherlands/219/2003(H7N7), and A/New York/107/2003(H7N2)] or with human A(H1N1)pdm09 (A/California/07/2009-like) or A(H3N2) (A/Perth16/2009) viruses. Mice produced equivalent titers of antibodies to all viruses as measured by enzyme-linked immunosorbent assay (ELISA). However, the antibody titers induced by H7 viruses were significantly lower when measured by HI and MN assays. Despite inducing very low levels of nAb, H7 vaccines conferred complete protection against homologous virus challenge in mice, and the serum antibodies directed against the HA head region were capable of mediating protection. The apparently low immunogenicity associated with H7 viruses and vaccines may be at least partly related to measuring antibody titers with the traditional HI and MN assays, which may not provide a true measure of protective immunity associated with H7 immunization. This study underscores the need for development of additional correlates of protection for prepandemic vaccines.IMPORTANCE H7 avian influenza viruses present a serious risk to human health. Preparedness efforts include development of prepandemic vaccines. For seasonal influenza viruses, protection is correlated with antibody

  11. Precisely Molded Nanoparticle Displaying DENV-E Proteins Induces Robust Serotype-Specific Neutralizing Antibody Responses

    PubMed Central

    Hoekstra, Gabriel; Yi, Xianwen; Stone, Michelle; Horvath, Katie; Miley, Michael J.; DeSimone, Joseph; Luft, Chris J.; de Silva, Aravinda M.

    2016-01-01

    Dengue virus (DENV) is the causative agent of dengue fever and dengue hemorrhagic fever. The virus is endemic in over 120 countries, causing over 350 million infections per year. Dengue vaccine development is challenging because of the need to induce simultaneous protection against four antigenically distinct DENV serotypes and evidence that, under some conditions, vaccination can enhance disease due to specific immunity to the virus. While several live-attenuated tetravalent dengue virus vaccines display partial efficacy, it has been challenging to induce balanced protective immunity to all 4 serotypes. Instead of using whole-virus formulations, we are exploring the potentials for a particulate subunit vaccine, based on DENV E-protein displayed on nanoparticles that have been precisely molded using Particle Replication in Non-wetting Template (PRINT) technology. Here we describe immunization studies with a DENV2-nanoparticle vaccine candidate. The ectodomain of DENV2-E protein was expressed as a secreted recombinant protein (sRecE), purified and adsorbed to poly (lactic-co-glycolic acid) (PLGA) nanoparticles of different sizes and shape. We show that PRINT nanoparticle adsorbed sRecE without any adjuvant induces higher IgG titers and a more potent DENV2-specific neutralizing antibody response compared to the soluble sRecE protein alone. Antigen trafficking indicate that PRINT nanoparticle display of sRecE prolongs the bio-availability of the antigen in the draining lymph nodes by creating an antigen depot. Our results demonstrate that PRINT nanoparticles are a promising platform for delivering subunit vaccines against flaviviruses such as dengue and Zika. PMID:27764114

  12. Mumps-specific cross-neutralization by MMR vaccine-induced antibodies predicts protection against mumps virus infection.

    PubMed

    Gouma, Sigrid; Ten Hulscher, Hinke I; Schurink-van 't Klooster, Tessa M; de Melker, Hester E; Boland, Greet J; Kaaijk, Patricia; van Els, Cécile A C M; Koopmans, Marion P G; van Binnendijk, Rob S

    2016-07-29

    Similar to other recent mumps genotype G outbreaks worldwide, most mumps patients during the recent mumps genotype G outbreaks in the Netherlands had received 2 doses of measles, mumps and rubella (MMR) vaccine during childhood. Here, we investigate the capacity of vaccine-induced antibodies to neutralize wild type mumps virus strains, including mumps virus genotype G. In this study, we tested 105 pre-outbreak serum samples from students who had received 2 MMR vaccine doses and who had no mumps virus infection (n=76), symptomatic mumps virus infection (n=10) or asymptomatic mumps virus infection (n=19) during the mumps outbreaks. In all samples, mumps-specific IgG concentrations were measured by multiplex immunoassay and neutralization titers were measured against the Jeryl Lynn vaccine strain and against wild type genotype G and genotype D mumps virus strains. The correlation between mumps-specific IgG concentrations and neutralization titers against Jeryl Lynn was poor, which suggests that IgG concentrations do not adequately represent immunological protection against mumps virus infection by antibody neutralization. Pre-outbreak neutralization titers in infected persons were significantly lower against genotype G than against the vaccine strain. Furthermore, antibody neutralization of wild type mumps virus genotype G and genotype D was significantly reduced in pre-outbreak samples from infected persons as compared with non-infected persons. No statistically significant difference was found for the vaccine strain. The sensitivity/specificity ratio was largest for neutralization of the genotype G strain as compared with the genotype D strain and the vaccine strain. The reduced neutralization of wild type mumps virus strains in MMR vaccinated persons prior to infection indicates that pre-outbreak mumps virus neutralization is partly strain-specific and that neutralization differs between infected and non-infected persons. Therefore, we recommend the use of wild

  13. The heptide repeat 2 and upstream region of TGEV induces potent cross-neutralizing antibodies against group I coronaviruses.

    PubMed

    Shi, Huiling; Wu, Nannan; Wang, Xiaoming; Wang, Tianhou

    2012-10-01

    The coronavirus heptide repeat (HR) region in the spike protein induces neutralizing antibodies that block the postfusion core formation and inhibit virus entry into target cells. The HR2 regions for coronaviruses of the same serogroup share high homology. We found that polyclonal antibodies derived from transmissible gastroenteritis coronavirus HR2 and upstream region were cross-reactive with the S proteins of the same serogroup in western blotting. The polyclonal antibodies also potently cross-neutralized viruses from the same serogroup. This study provides new insight for designing vaccine and therapeutic reagents against coronavirus infections.

  14. A measurement of neutrino-induced charged-current neutral pion production

    NASA Astrophysics Data System (ADS)

    Nelson, Robert H.

    This work presents the first comprehensive measurement of neutrino-induced charged-current neutral pion production (CCpi0) off a nuclear target. The Mini Booster Neutrino Experiment (MiniBooNE) and Booster Neutrino Beam (BNB) are discussed in detail. MiniBooNE is a high-statistics (˜1,000,000 interactions) low-energy (Enu ∈ 0.5--2.0 GeV) neutrino experiment located at Fermilab. The method for selecting and reconstructing CCpi0 events is presented. The pi0 and mu - are fully reconstructed in the final state allowing for the measurement of, among other things, the neutrino energy. The total observable CCpi 0 cross-section is presented as a function of neutrino energy, along with five differential cross-sections in terms of the final state kinematics and Q2. The results are combined to yield a flux-averaged total cross-section of phi = (9.2 +/- 0.3stat. +/- 1.5 syst.) x 10--39 cm 2/CH2 at energy 965 MeV. These measurements will aid future neutrino experiments with the prediction of their neutrino interaction rates.

  15. Weak charged and neutral current induced one pion production off the nucleon

    NASA Astrophysics Data System (ADS)

    Rafi Alam, M.; Sajjad Athar, M.; Chauhan, S.; Singh, S. K.

    2016-02-01

    We present a study of neutrino/antineutrino induced charged current (CC) and neutral current (NC) single pion production (SPP) off the nucleon. For this, we have considered P33(1232) resonance, nonresonant background (NRB) terms, other higher resonances like P11(1440), S11(1535), D13(1520), S11(1650) and P13(1720). For the NRB terms a microscopic approach based on SU(2) nonlinear sigma model has been used. The vector form factors for the resonances are obtained by using the relationship between the electromagnetic resonance form factors and helicity amplitudes provided by MAID. Axial coupling C5A(0) in the case of P33(1232) resonance is obtained by fitting the ANL and BNL ν-deuteron reanalyzed scattering data. The results are presented with and without deuteron effect for the total scattering cross-sections for all possible channels, viz. νl(ν¯l)+N → l-(l+)+N‧+πi and νl(ν¯l)+N → νl(ν¯l)+N‧ + πi, where N,N‧ = p,n, πi = π± or π0 and l = e,μ.

  16. A Measurement of Neutrino-Induced Charged-Current Neutral Pion Production

    SciTech Connect

    Nelson, Robert H.

    2010-01-01

    This work presents the first comprehensive measurement of neutrino-induced charged-current neutral pion production (CCπ0) off a nuclear target. The Mini Booster Neutrino Experiment (MiniBooNE) and Booster Neutrino Beam (BNB) are discussed in detail. MiniBooNE is a high-statistics (~ 1, 000, 000 interactions) low-energy (Evϵ 2 0.5 - 2.0 GeV) neutrino experiment located at Fermilab. The method for selecting and reconstructing CCπ0 events is presented. The π0 and μ- are fully reconstructed in the final state allowing for the measurement of, among other things, the neutrino energy. The total observable CCπ0 cross-section is presented as a function of neutrino energy, along with five differential cross-sections in terms of the final state kinematics and Q2. The results are combined to yield a flux-averaged total cross-section of <σ>Φ = (9.2 ± 0.3stat. ± 1.5syst.) × 10-39 cm2/CH2 at energy 965 MeV. These measurements will aid future neutrino experiments with the prediction of their neutrino interaction rates.

  17. Virtual gap states induced modifications in charge neutrality level in cadmium oxide thin films

    NASA Astrophysics Data System (ADS)

    Das, Arkaprava; Gautam, Subodh K.; Umapathy, G. R.; Ojha, S.; Singh, Fouran

    2017-04-01

    Present manuscript reports on the creation of virtual gap states (ViGS) in sol-gel spin coated cadmium oxide thin films by tin doping and irradiation at different density of electronic excitations (EEs) induced by energetic ions. Effects of ViGS on charge neutrality level (CNL), which affects the modifications in band gap (BG) of such oxide semiconductors are investigated. Studies reveal that there were insignificant changes in the structural and morphological properties of the films. Interestingly, the transmittance spectrum shows insignificant changes in the films irradiated by low density EEs but an overall decrease in transmittance for high density EEs. However, Raman spectroscopy reveals a significant change in longitudinal optical (LO) modes with tin doping which get intensified and they exhibit an insignificant influence upon irradiation. The BG with respect to bulk is observed to be increased by 0.57 eV and 0.83 eV upon tin doping and irradiation, respectively. These variations are attributed to the combined effect of Burstein-Moss shift (BMS) and ViGS, which were created by tin doping and EEs. It is observed that density of ViGS strongly depends on the density of EEs. A schematic band diagram is also developed for demonstrating the observed influence of ViGS on band gap modifications. Thus the reported investigations are very interesting for the in-depth understanding of fundamental interactions and besides their possible potential applications in the development of optoelectronic devices.

  18. Fisheries-induced neutral and adaptive evolution in exploited fish populations and consequences for their adaptive potential

    PubMed Central

    Marty, Lise; Dieckmann, Ulf; Ernande, Bruno

    2015-01-01

    Fishing may induce neutral and adaptive evolution affecting life-history traits, and molecular evidence has shown that neutral genetic diversity has declined in some exploited populations. Here, we theoretically study the interplay between neutral and adaptive evolution caused by fishing. An individual-based eco-genetic model is devised that includes neutral and functional loci in a realistic ecological setting. In line with theoretical expectations, we find that fishing induces evolution towards slow growth, early maturation at small size and higher reproductive investment. We show, first, that the choice of genetic model (based on either quantitative genetics or gametic inheritance) influences the evolutionary recovery of traits after fishing ceases. Second, we analyse the influence of three factors possibly involved in the lack of evolutionary recovery: the strength of selection, the effect of genetic drift and the loss of adaptive potential. We find that evolutionary recovery is hampered by an association of weak selection differentials with reduced additive genetic variances. Third, the contribution of fisheries-induced selection to the erosion of functional genetic diversity clearly dominates that of genetic drift only for the traits related to maturation. Together, our results highlight the importance of taking into account population genetic variability in predictions of eco-evolutionary dynamics. PMID:25667602

  19. Fisheries-induced neutral and adaptive evolution in exploited fish populations and consequences for their adaptive potential.

    PubMed

    Marty, Lise; Dieckmann, Ulf; Ernande, Bruno

    2015-01-01

    Fishing may induce neutral and adaptive evolution affecting life-history traits, and molecular evidence has shown that neutral genetic diversity has declined in some exploited populations. Here, we theoretically study the interplay between neutral and adaptive evolution caused by fishing. An individual-based eco-genetic model is devised that includes neutral and functional loci in a realistic ecological setting. In line with theoretical expectations, we find that fishing induces evolution towards slow growth, early maturation at small size and higher reproductive investment. We show, first, that the choice of genetic model (based on either quantitative genetics or gametic inheritance) influences the evolutionary recovery of traits after fishing ceases. Second, we analyse the influence of three factors possibly involved in the lack of evolutionary recovery: the strength of selection, the effect of genetic drift and the loss of adaptive potential. We find that evolutionary recovery is hampered by an association of weak selection differentials with reduced additive genetic variances. Third, the contribution of fisheries-induced selection to the erosion of functional genetic diversity clearly dominates that of genetic drift only for the traits related to maturation. Together, our results highlight the importance of taking into account population genetic variability in predictions of eco-evolutionary dynamics.

  20. Effect on extrapulmonary sepsis-induced acute lung injury by hemoperfusion with neutral microporous resin column.

    PubMed

    Huang, Zhao; Wang, Si-rong; Yang, Zi-li; Liu, Ji-yun

    2013-08-01

    The aim of this study was to investigate the effect of neutral microporous resin hemoperfusion on oxygenation improvement, removal of inflammatory cytokines in plasma and bronchoalveolar lavage, and mortality in acute lung injury induced by extrapulmonary sepsis. Forty-six patients with acute lung injury induced by extrapulmonary sepsis were randomized to HA type hemoperfusion treatment (N=25) or standard therapy (N=21). Those undergoing hemoperfusion treatment received HA330 hemoperfusion. We measured the plasma and bronchoalveolar lavage concentrations of TNF-α and IL-1, and the following parameters were compared between the control group and the hemoperfusion group on days 0, 3 and 7: lung injury measurements (arterial oxygen tension/fractional inspired oxygen ratio, lung injury score, chest X-ray score); interstitial edema of lung (extravascular lung water). Duration of mechanical ventilation, hospital, 28-day, and intensive care unit mortality were also observed. Patients treated with HA hemoperfusion showed a significant removal of plasma and bronchoalveolar lavage TNF-α and IL-1 over time while in the study. Patients in the HA group also demonstrated not only significant improvement of PaO2 /FiO2 , but also decreased Lung Injury Score and chest X-ray score at days 3 and 7. Furthermore, the measurements of the arterial oxygen tension/fractional inspired oxygen ratio, lung injury score and extravascular lung water (EVLWI) significantly correlated with and the concentration of cytokines in the plasma (all P<0.05). The HA hemoperfusion treatment group had a significant reduction in duration of mechanical ventilation, length of intensive care unit stay, and intensive care unit mortality. Significant removal of inflammatory cytokines from circulation and lung by hemoperfusion treatment using the HA type cartridge may contribute to the improvement of lung injury and intensive care unit outcome in extrapulmonary septic patients. © 2012 The Authors. Therapeutic

  1. Neutralizing and cross-neutralizing antibody titres induced by bivalent and quadrivalent human papillomavirus vaccines in the target population of organized vaccination programmes.

    PubMed

    Barzon, Luisa; Squarzon, Laura; Masiero, Serena; Pacenti, Monia; Marcati, Giorgia; Mantelli, Barbara; Gabrielli, Liliana; Pascucci, Maria Grazia; Lazzarotto, Tiziana; Caputo, Antonella; Palù, Giorgio

    2014-09-15

    Aim of this investigator-initiated study was to evaluate and compare the titres of neutralizing and cross-neutralizing antibodies (NAbs) induced by the bivalent (Cervarix(®)) and quadrivalent (Gardasil(®)) HPV vaccines in a cohort of girls aged 11-13 years from organized vaccination programmes. To this aim, HPV16 and HPV18 NAbs were measured by pseudovirion-based neutralization assays in serum collected at 1-6 months after the third vaccine dose in 107 girls vaccinated with Cervarix(®) and 126 vaccinated with Gardasil(®), while HPV31 and HPV45 cross-NAbs were tested in the first 50 consecutive girls of both vaccine groups. The results of this study demonstrated that all vaccinated girls developed HPV16 and HPV18 NAbs, with the exception of two Gardasil(®) vaccinees with undetectable HPV18 NAbs. Geometric mean titres (GMTs) of both HPV16 and HPV18 NAbs were significantly higher in Cervarix(®) than in Gardasil(®) vaccinees [HPV16 NAb GMT 22,136 (95% CI, 18,811-26,073) vs 5092 (4230-6151), respectively; P<0.0001; HPV18 NAb GMT 11,962 (9536-14,363) vs 1804 (1574-2110), respectively; P<0.0001]. Cross-NAbs to HPV31 and HPV45 were detected more frequently Cervarix(®) (HPV31 NAb positivity rates 92.7% and 36%, respectively; P<0.05) than in Gardasil(®) vaccinees (HPV45 NAb positivity rates 56% and 6%, respectively; P<0.0001). The titres of cross-NAbs against HPV31 and HPV45 were also significantly higher in Cervarix(®) than in Gardasil(®) vaccinees [HPV31 NAb GMT 157.2 (95% CI, 92-269) vs 13.0 (6.5-25.8), respectively; P<0.0001; HPV45 NAb GMT 4.7 (2.1-10.2) vs 1.3 (0.3-3.1), respectively; P<0.01]. In conclusion, in adolescent girls vaccinated within organized vaccination programmes, HPV vaccines drive the generation not only of NAbs to HPV vaccine types, but also of cross-NAbs. The bivalent vaccine induced significantly higher HPV16 and HPV18 NAb titres and more frequently and at higher titre HPV31 and HPV45 cross-NAbs than the quadrivalent vaccine. Copyright

  2. TGF-β Neutralization Enhances AngII-Induced Aortic Rupture and Aneurysm in Both Thoracic and Abdominal Regions

    PubMed Central

    Howatt, Deborah A.; Balakrishnan, Anju; Moorleghen, Jessica J.; Cassis, Lisa A.; Daugherty, Alan

    2016-01-01

    AngII and TGF-β interact in development of thoracic and abdominal aortic diseases, although there are many facets of this interaction that have not been clearly defined. The aim of the present study was to determine the effects of TGF-β neutralization on AngII induced-aortic pathologies. Male C57BL/6J mice were administered with either a rabbit or mouse TGF-β neutralizing antibody and then infused with AngII. The rabbit TGF-β antibody modestly reduced serum TGF-β concentrations, with no significant enhancements to AngII-induced aneurysm or rupture. Administration of this rabbit TGF-β antibody in mice led to high serum titers against rabbit IgG that may have attenuated the neutralization. In contrast, a mouse TGF-β antibody (1D11) significantly increased rupture in both the ascending and suprarenal aortic regions, but only at doses that markedly decreased serum TGF-β concentrations. High doses of 1D11 antibody significantly increased AngII-induced ascending and suprarenal aortic dilatation. To determine whether TGF-β neutralization had effects in mice previously infused with AngII, the 1D11 antibody was injected into mice that had been infused with AngII for 28 days and were observed during continued infusion for a further 28 days. Despite near ablations of serum TGF-β concentrations, the mouse TGF-β antibody had no effect on aortic rupture or dimensions in either ascending or suprarenal region. These data provide further evidence that AngII-induced aortic rupture is enhanced greatly by TGF-β neutralization when initiated before pathogenesis. PMID:27104863

  3. Laser Induced Fluorescence Spectroscopy of Neutral and Ionized Polycyclic Aromatic Hydrocarbons in a Cosmic Simulation Chamber

    NASA Astrophysics Data System (ADS)

    Bejaoui, Salma; Salama, Farid

    2015-08-01

    Polycyclic aromatic hydrocarbon (PAH) molecules are considered the best carriers to account for the ubiquitous infrared emission bands. PAHs have also been proposed as candidates to explain the diffuse interstellar bands (DIBs), a series of absorption features seen on the interstellar extinction curve and are plausible carriers for the extended red emission (ERE), a photoluminescent process associated with a wide variety of interstellar environments. Extensive efforts have been devoted over the past two decades to characterize the physical and chemical properties of PAH molecules and ions in space. Absorption spectra of PAH molecules and ions trapped in solid matrices have been compared to the DIBs [1, 2]. Absorption spectra of several cold, isolated gas-phase PAHs have also been measured under experimental conditions that mimic the interstellar conditions [see 3 for a review]. The purpose of this study is to provide a new dimension to the existing spectroscopic database of neutral and single ionized PAHs that is largely based on absorption spectra by adding emission spectroscopy data. The measurements are based on the laser-induced fluorescence (LIF) technique [4] and are performed with the Pulsed Discharge Nozzle (PDN) of the COSmIC laboratory facility at NASA Ames laboratory. The PDN generates plasma in a free supersonic jet expansion to simulate the physical and the chemical conditions in interstellar environments. We focus, here, on the fluorescence spectra of large neutral PAHs and their cations where there is a lack of fluorescence spectroscopy data. The astronomical implications of the data (e.g., ERE) are examinedReferences[1] F. Salama, E. Bakes, L.J. Allamandola, A.G.G.M. Tielens, Astrophys. J., 458 (1996) p.621[2] F. Salama, The ISO Revolution, EDP Sciences, Les Ulis, France (1999) p.65[3] Salama F., In Organic Matter in Space, IAU Symposium 251, Kwok & Sandford Eds.Cambridge University Press,4, S251,(2008), p. 357 (2008) and references therein.[4

  4. Biology and management of Longidorus americanus in a southern USA nursery

    Treesearch

    M.M. Cram; S.W. Fraedrich

    2007-01-01

    A recently discovered nematode, Longidorus americanus, caused stunting of Pinus taeda seedlings at a forest-tree nursery in Georgia, USA. In growth chamber experiments, L. americanus significantly reduced the root size of I? taeda and P elliottii seedlings. Although the root systems of L. palustris...

  5. Muscular Sarcocystis infection in a bear (Ursus americanus).

    PubMed

    Dubey, J P; Topper, M J; Nutter, F B

    1998-04-01

    Sarcocysts of an unidentified Sarcocystis species were found in sections of skeletal muscles of a black bear (Ursus americanus) from North Carolina. Two sarcocysts in a section measured 45 x 37.5 microm and 67.5 x 50 microm and had a thin (<2 microm) sarcocyst wall. The villar protrusions on the cyst wall were up to 2 microm long and up to 0.7 microm wide. The bradyzoites were approximately 6 X 2.5 microm in size. This is the first report of muscular Sarcocystis in a bear.

  6. Migrations and growth of deep-sea lobsters, Homarus americanus.

    PubMed

    Cooper, R A; Uzmann, J R

    1971-01-22

    In distinct contrast to the restricted movements of coastal stocks of lobsters (Homarus americanus), those inhabiting the outer continental shelf undertake extensive seasonal migrations. Of 5710 tagged lobsters released on the outer continental shelf off New England from April 1968 to June 1969, 400 had been recaptured by April 1970. The distribution of the recoveries demonstrated shoalward migration in spring and summer and a return to the edge of the shelf in fall and winter. Deep-sea lobsters have a faster rate of growth than coastal lobsters; growth increments at molting and the frequency of molting are greater.

  7. An oral Aujeszky's disease vaccine (YS-400) induces neutralizing antibody in pigs

    PubMed Central

    2016-01-01

    Purpose Aujeszky's disease (AD) is an economically important disease affecting both wild and domestic pigs of the species Sus scrofa. A previous study yielded serological evidence of AD in Korean wild boars, which could spread AD to other animals. A new Aujeszky's disease virus (ADV) bait vaccine is required to prevent AD outbreaks in swine. In the present study, we investigated the safety and immunogenicity of a gE-deleted marker vaccine, strain YS-400, in young domestic pigs. Materials and Methods The YS-400 strain was propagated in Vero cells, and the trial ADV bait vaccine (a vaccine blister in a matrix including an attractant) was prepared. Pigs were orally immunized with the vaccine (2 mL, 107.5 TCID50/mL) delivered using a syringe or in the bait vaccine. The animals were observed for 9 weeks after vaccination, and immunogenicity was assessed using a virus neutralization (VN) test and enzyme linked immunosorbent assay. Results The YS-400 strain was non-pathogenic to pigs when given orally and induced high VN titers (1:32-1:128) 6 weeks post-administration. Of the pigs given the ADV bait vaccine twice or three times, 40% were seropositive by 2 weeks, and 100% were seropositive by 7 weeks after the first dose. Pigs that consumed the AD bait vaccine three times developed VN titers that were slightly higher than those of pigs given the vaccine twice. Conclusion Domestic pigs given the trial ADV bait vaccine exhibited no adverse effects and developed high VN titers against ADV, indicating that the YS-400 strain is safe and can prevent ADV infection in domestic pigs. PMID:27489803

  8. Variability in detection and quantification of interferon β-1b–induced neutralizing antibodies

    PubMed Central

    2012-01-01

    Background Interferon-beta (IFNB) therapy for multiple sclerosis can lead to the induction of neutralizing antibodies (NAbs) against IFNB. Various methods are used for detection and quantification of NAbs. Methods Blood samples from 125 IFNB-1b–treated patients, which were tested NAb negative or NAb positive after conclusion of a clinical study, were retested three years after first being assessed in four different laboratories that offer routine NAb testing to practicing neurologists. The myxovirus protein A (MxA) induction assay, the cytopathic effect (CPE) assay (two laboratories), or the luciferase assay were used. Intra- and inter-laboratory agreement between assays with respect to NAb detection and NAb titer quantification were evaluated. Results High agreement for NAb detection (kappa coefficient, 0.86) and for titer levels was observed for the intra-laboratory comparison in the laboratory using the MxA induction assay performed three years ago and now. A similarly high agreement for NAb detection (kappa coefficient, 0.87) and for titer quantification was noted for the MxA assay of this laboratory with one of two laboratories using the CPE assay. All other inter-laboratory comparisons showed kappa values between 0.57 and 0.68 and remarkable differences in individual titer levels. Conclusions There are considerable differences in the detection and quantification of IFNB-induced NAbs among laboratories offering NAb testing for clinical practice using different assay methods. It is important that these differences are considered when interpreting NAb results for clinical decision-making and when developing general recommendations for potentially clinically meaningful NAb titer levels. PMID:22703536

  9. Goosefish Lophius americanus fecundity and spawning frequency, with implications for population reproductive potential.

    PubMed

    McBride, R S; Johnson, A K; Lindsay, E K; Walsh, H J; Richards, R A

    2017-02-23

    To improve knowledge of goosefish Lophius americanus' reproductive biology, females were collected during 2009-2012 from the Mid-Atlantic Bight shelf region of the U.S. east coast. Batch fecundity increased with total length (LT ), from 229 100 to 2 243 300 mature oocytes per female (LT range: 55·5-112 cm; n = 54). This estimate of fecundity at LT is lower than one derived from a sample collected during 1982-1985. Examination of whole oocyte diameters in different months indicated that L. americanus is a serial spawner, releasing more than one egg veil per spawning season, as suspected or observed for other Lophius species. Seasonality of spawning was evident from whole oocytes and gonad histology, and from larval fish surveys spanning the U.S. north-east shelf, and confirmed a protracted (c. 6 months) spawning period. Peak spawning activity progressed northward from spring to autumn. The population-level implications of these results were explored by estimating population reproductive potential (PRP ), which considered the value of both current and future per capita reproduction using decade-specific age structure and fecundity at length. PRP is now more than 50% lower compared with the historical period (1982-1985), a result of the lower proportions of large females and reduced fecundity across all sizes. Mechanisms that could explain this loss of stock productivity are fishing-induced size-age truncation or regime shifts in egg production caused by changes in energy density of common forage species.

  10. Measurement of muon neutrino and antineutrino induced single neutral pion production cross sections

    SciTech Connect

    Anderson, Colin E.

    2011-05-01

    Elucidating the nature of neutrino oscillation continues to be a goal in the vanguard of the efforts of physics experiment. As neutrino oscillation searches seek an increasingly elusive signal, a thorough understanding of the possible backgrounds becomes ever more important. Measurements of neutrino-nucleus interaction cross sections are key to this understanding. Searches for νμ → νe oscillation - a channel that may yield insight into the vanishingly small mixing parameter θ13, CP violation, and the neutrino mass hierarchy - are particularly susceptible to contamination from neutral current single π0 (NC 1π0) production. Unfortunately, the available data concerning NC 1π0 production are limited in scope and statistics. Without satisfactory constraints, theoretical models of NC 1π0 production yield substantially differing predictions in the critical Eν ~ 1 GeV regime. Additional investigation of this interaction can ameliorate the current deficiencies. The Mini Booster Neutrino Experiment (MiniBooNE) is a short-baseline neutrino oscillation search operating at the Fermi National Accelerator Laboratory (Fermilab). While the oscillation search is the principal charge of the MiniBooNE collaboration, the extensive data (~ 106 neutrino events) offer a rich resource with which to conduct neutrino cross section measurements. This work concerns the measurement of both neutrino and antineutrino NC 1π0 production cross sections at MiniBooNE. The size of the event samples used in the analysis exceeds that of all other similar experiments combined by an order of magnitude. We present the first measurements of the absolute NC 1π0 cross section as well as the first differential cross sections in both neutrino and antineutrino mode. Specifically, we measure single differential cross sections with respect to pion momentum and pion angle. We find the

  11. On the cosmic-ray induced background in neutral pion production measurements with a BaF 2 multidetector

    NASA Astrophysics Data System (ADS)

    Badalà, A.; Barbera, R.; Palmeri, A.; Pappalardo, G. S.; Riggi, F.; Russo, A. C.; Russo, G.; Turrisi, R.

    1995-02-01

    The problem of the cosmic-ray induced background in neutral pion production measurements with barium fluoride multidetectors is discussed. As a reference example, the response to cosmic rays of the MEDEA photon spectrometer is studied. The interaction of the cosmic radiation with the experimental filter has been treated by means of full Monte Carlo computer simulations with the GEANT3 code. The results of the simulations are compared with experimental data and general criteria to minimize the cosmic background are discussed.

  12. An interleukin-6-neutralizing antibody prevents cyclosporine-induced nephrotoxicity in mice.

    PubMed

    LaSpina, Mark; Tripathi, Sudipta; Gatto, Louis A; Bruch, David; Maier, Kristopher G; Kittur, Dilip S

    2008-08-01

    observed to have 15% (6/40) damage. Comparing imaging analysis, there was no difference between mice treated with CyA alone and with CyA with apocynin. However, the amount of damage in mice treated with CyA and IL-6 antibody was found to be significantly lower than both CyA and CyA with apocynin. CyA action as a calcineurin inhibitor has allowed prolongation of kidney transplants, but its chronic use has led to devastating consequences such as allograft nephropathy. Previously, we have identified potential mechanisms of CyA-induced endothelial dysfunction in vitro. The current study identifies increased IL-6 expression as a mechanism by which CyA induces renal damage and that the use of an IL-6-neutralizing antibody may be useful in reducing CyA-induced renal damage.

  13. Neutralization of MMP-2 protects Staphylococcus aureus infection induced septic arthritis in mice and regulates the levels of cytokines.

    PubMed

    Sultana, Sahin; Adhikary, Rana; Nandi, Ajeya; Bishayi, Biswadev

    2016-10-01

    Matrix metalloproteinases (MMPs) are crucial players in Staphylococcus aureus mediated synovial tissue destruction in the pathogenesis of septic arthritis. Bacterial insult increases proteolytic matrix fragments by activated chondrocytes and synovial fibroblasts leading to induction of matrix metalloproteinases. Tissue destruction via MMPs induced by bacterial products, necrotic tissues and proinflammatory cytokines have been reported. Cytokines like TNF-α, IL-1β released from host cells in response to S. aureus infection promote cartilage degradation by stimulating the production of MMPs. Antibiotic treatment can eradicate invading bacteria but elevated levels of cytokines and cytokines induced MMPs activation lead to progressive and devastating bone and cartilage destruction even after bacterial clearance. Like other MMPs, MMP-2 also contributes to extracellular matrix degradation in different types of arthritis. Release of certain pro inflammatory cytokines can also be regulated by MMP-2 activation leading to further tissue destruction. The role of MMP-2 in the pathogenesis of S. aureus infection induced septic arthritis and its influence on cytokines regulation needs further investigation. Whether neutralization of MMP-2 provides protection against Staphylococcus aureus infection induced septic arthritis in mice is an obvious question. Here we reported that neutralization of MMP-2 during S. aureus infection induced septic arthritis might be beneficial for preventing infection induced extracellular matrix destruction thereby decreasing bacterial burden in synovial tissues and regulating inflammatory cytokines in arthritic mice. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. Stress-Induced Sphingolipid Signaling: Role of Type-2 Neutral Sphingomyelinase in Murine Cell Apoptosis and Proliferation

    PubMed Central

    Devillard, Raphael; Galvani, Sylvain; Thiers, Jean-Claude; Guenet, Jean-Louis; Hannun, Yusuf; Bielawski, Jacek; Nègre-Salvayre, Anne; Salvayre, Robert; Augé, Nathalie

    2010-01-01

    Background Sphingomyelin hydrolysis in response to stress-inducing agents, and subsequent ceramide generation, are implicated in various cellular responses, including apoptosis, inflammation and proliferation, depending on the nature of the different acidic or neutral sphingomyelinases. This study was carried out to investigate whether the neutral Mg2+-dependent neutral sphingomyelinase-2 (nSMase2) plays a role in the cellular signaling evoked by TNFalpha and oxidized LDLs, two stress-inducing agents, which are mitogenic at low concentrations and proapoptotic at higher concentrations. Methodology and Principal Findings For this purpose, we used nSMase2-deficient cells from homozygous fro/fro (fragilitas ossium) mice and nSMase2-deficient cells reconstituted with a V5-tagged nSMase2. We report that the genetic defect of nSMase2 (in fibroblasts from fro/fro mice) does not alter the TNFalpha and oxidized LDLs-mediated apoptotic response. Likewise, the hepatic toxicity of TNFalpha is similar in wild type and fro mice, thus is independent of nSMase2 activation. In contrast, the mitogenic response elicited by low concentrations of TNFalpha and oxidized LDLs (but not fetal calf serum) requires nSMase2 activation. Conclusion and Significance nSMase2 activation is not involved in apoptosis mediated by TNFalpha and oxidized LDLs in murine fibroblasts, and in the hepatotoxicity of TNFalpha in mice, but is required for the mitogenic response to stress-inducing agents. PMID:20352118

  15. Hookworm (Necator americanus) larval enzymes disrupt human vascular endothelium.

    PubMed

    Souadkia, Nahed; Brown, Alan; Leach, Lopa; Pritchard, David I

    2010-09-01

    Knowledge of the molecular mechanisms used by Necator americanus larvae to penetrate the human skin and the vasculature would aid the development of effective vaccines against this important pathogen. In this work, the impact of N. americanus exsheathing fluid (EF) and excretory/secretory products (ES) on the endothelial barrier was examined using human umbilical vein endothelial cells (HUVEC). Cellular responses were assessed by investigating molecular changes at cell-cell junctions and by determining levels of secreted IL-6, IL-8, and vascular endothelial growth factor (VEGF) in the culture medium. It would appear that a repertoire of larval proteases caused a dose-related increase in endothelial permeability as characterized by a decrease in monolayer resistance with increased permeation of tracer-albumin. These barrier changes were associated with disruption of junctional vascular endothelial cadherin (VE-cadherin) and F-actin and an increase in endothelial secretion of IL-6 and IL-8. Our data suggest that larval proteases play an important role in negotiating the endothelium.

  16. Exploring the Microbial Community of Shell Disease in Homarus americanus

    NASA Astrophysics Data System (ADS)

    Unzueta Martinez, A.; Feinman, S. G.; Bowen, J. L.; Tlusty, M. F.

    2016-02-01

    The American lobster, Homarus americanus is an important commercial fishery in New England, and generated nearly half a billion dollars in revenue in 2012. Outbreaks of shell disease among H. americanus populations have caused a great deal of concern regarding its cause and spread. Although shell disease is not usually lethal, afflicted lobsters have an unattractive appearance and are generally unmarketable. Shell disease is hypothesized to be caused by infectious microorganisms, but no etiological agent has been identified. This project characterizes the microbial communities associated with the formation of shell disease in lobsters and how these communities change during the progression of the disease. Microbial samples were collected on lobsters at the site of a new shell disease location, as well as at 0.5, 1, and 1.5 cm away from the site. Microbial DNA was extracted from each sample, the 16S rRNA gene amplified using PCR, and sequenced using high throughput sequencing. The results showed that microbial communities on diseased lobster shells were significantly different spatially, but not temporally. The diversity of microbes was lowest at the site of shell disease, and increased with distance from the site. This work elucidated the microbial community composition of shell disease, tracked this community throughout the progression of disease, and demonstrated a gradual rather than abrupt demarcation in microbial community between symptomatic and asymptomatic shell.

  17. Laser Induced Fluorescence (LIF) Data for Neutral Argon (ArI) in a Large Scale Helicon Plasm

    NASA Astrophysics Data System (ADS)

    Kelly, Ralph; Gilmore, Mark; Meany, Kevin; Zhang, Yue; Desjardins, Tiffany

    2016-10-01

    When neutral and ion densities are spatially non-uniform, neutral-ion collisions can exert a torque on a magnetized plasma column via the FxB force, where F is the force exerted on ions by neutrals. This FxB force may have a significant effect on the dynamics of plasma instabilities and flows. In order to investigate the role of neutral dynamics in helicon discharges in the HelCat (Helicon-Cathode) plasma device at U. New Mexico, an Ar I Laser Induced Fluorescence (LIF) system has been developed. The LIF system is based on a >250 mW, tunable solid state laser. The laser will pumps the metastable (2P3/2 0) 4s level to the (2P1/2 0) 4p level using 696. 7352 nm light, and fluorescence radiation from decay to the (2P1/2 0) 4s level at 772. 6333 nm is observed. The system design and initial data will be presented.

  18. Vortex-induced vibrations of a neutrally buoyant circular cylinder near a plane wall

    NASA Astrophysics Data System (ADS)

    Wang, X. K.; Hao, Z.; Tan, S. K.

    2013-05-01

    This paper presents an experimental study of the motions, drag force and vortex shedding patterns of an elastically mounted circular cylinder, which is held at various heights above a plane wall and is subject to vortex-induced vibration (VIV) in the transverse direction. The cylinder is neutrally buoyant with a mass ratio m=1.0 and has a low damping ratio ζ=0.0173. Effects of the gap ratio (S/D) ranged from 0.05 to 2.5 and the free-stream velocity (U) ranged from 0.15 to 0.65m/s (corresponding to 3000≤Re≤13 000, and 1.53≤U≤6.62) are examined. The flow around the cylinder has been measured using particle image velocimetry (PIV), in conjunction with direct measurements of the dynamic drag force on the cylinder using a piezoelectric load cell. Results of the vibrating cylinder under unbounded (or free-standing) condition, as well as those of a near-wall stationary cylinder at the same gap ratios, are also provided. For the free-standing cylinder, the transition from the initial branch to the upper branch is characterized by a switch of vortex pattern from the classical 2S mode to the newly-discovered 2PO mode by Morse and Williamson (2009). The nearby wall not only affects the amplitude and frequency of vibration, but also leads to non-linearities in the cylinder response as evidenced by the presence of super-harmonics in the drag force spectrum. In contrast to the case of a stationary cylinder that vortex shedding is suppressed below a critical gap ratio (S/D≈0.3), the elastically mounted cylinder always vibrates even at the smallest gap ratio S/D=0.05. Due to the proximity of the plane wall, the vortices shed from the vibrating cylinder that would otherwise be in a double-sided vortex street pattern (either 2S or 2PO mode) under free-standing condition are arranged into a single-sided pattern.

  19. Neutralization of interleukin-17A delays progression of silica-induced lung inflammation and fibrosis in C57BL/6 mice

    SciTech Connect

    Chen, Ying; Li, Cuiying; Weng, Dong; Song, Laiyu; Tang, Wen; Dai, Wujing; Yu, Ye; Liu, Fangwei; Zhao, Ming; Lu, Chunwei; Chen, Jie

    2014-02-15

    Silica exposure can cause lung inflammation and fibrosis, known as silicosis. Interleukin-17A (IL-17A) and Th17 cells play a pivotal role in controlling inflammatory diseases. However, the roles of IL-17A and Th17 cells in the progress of silica-induced inflammation and fibrosis are poorly understood. This study explored the effects of IL-17A on silica-induced inflammation and fibrosis. We used an anti-mouse IL-17A antibody to establish an IL-17A-neutralized mice model, and mice were exposed to silica to establish an experimental silicosis model. We showed that IL-17A neutralization delayed neutrophil accumulation and progression of silica-induced lung inflammation and fibrosis. IL-17A neutralization reduced the percentage of Th17 in CD4 + T cells, decreased IL-6 and IL-1β expression, and increased Tregs at an early phase of silica-induced inflammation. Neutralization of IL-17A delayed silica-induced Th1/Th2 immune and autoimmune responses. These results suggest that IL-17A neutralization alleviates early stage silica-induced lung inflammation and delays progression of silica-induced lung inflammation and fibrosis. Neutralization of IL-17A suppressed Th17 cell development by decreasing IL-6 and/or IL-1β and increased Tregs at an early phase of silica-induced inflammation. Neutralization of IL-17A also delayed the Th1/Th2 immune response during silica-induced lung inflammation and fibrosis. IL-17A may play a pivotal role in the early phase of silica-induced inflammation and may mediate the Th immune response to influence silica-induced lung inflammation and fibrosis in mice. - Highlights: • Neutralization of IL-17A alleviated silica-induced lung inflammation of early stage. • Neutralization of IL-17A decreased Th17 cells and increased Tregs. • IL-17A mediated the reciprocal relationship of Th17/Tregs by IL-6 and/or IL-1β. • Neutralization of IL-17A delayed silica-induced Th1/Th2 immune response. • Neutralization of IL-17A delayed silica-induced lung

  20. Hepatitis C Virus E1 and E2 Proteins Used as Separate Immunogens Induce Neutralizing Antibodies with Additive Properties

    PubMed Central

    Beaumont, Elodie; Roch, Emmanuelle; Chopin, Lucie; Roingeard, Philippe

    2016-01-01

    Various strategies involving the use of hepatitis C virus (HCV) E1 and E2 envelope glycoproteins as immunogens have been developed for prophylactic vaccination against HCV. However, the ideal mode of processing and presenting these immunogens for effective vaccination has yet to be determined. We used our recently described vaccine candidate based on full-length HCV E1 or E2 glycoproteins fused to the heterologous hepatitis B virus S envelope protein to compare the use of the E1 and E2 proteins as separate immunogens with their use as the E1E2 heterodimer, in terms of immunogenetic potential and the capacity to induce neutralizing antibodies. The specific anti-E1 and anti-E2 antibody responses induced in animals immunized with vaccine particles harboring the heterodimer were profoundly impaired with respect to those in animals immunized with particles harboring E1 and E2 separately. Moreover, the anti-E1 and anti-E2 antibodies had additive neutralizing properties that increase the cross-neutralization of heterologous strains of various HCV genotypes, highlighting the importance of including both E1 and E2 in the vaccine for an effective vaccination strategy. Our study has important implications for the optimization of HCV vaccination strategies based on HCV envelope proteins, regardless of the platform used to present these proteins to the immune system. PMID:26966906

  1. Neutralized ion beam modification of cellulose membranes for study of ion charge effect on ion-beam-induced DNA transfer

    NASA Astrophysics Data System (ADS)

    Prakrajang, K.; Sangwijit, K.; Anuntalabhochai, S.; Wanichapichart, P.; Yu, L. D.

    2012-02-01

    Low-energy ion beam biotechnology (IBBT) has recently been rapidly developed worldwide. Ion-beam-induced DNA transfer is one of the important applications of IBBT. However, mechanisms involved in this application are not yet well understood. In this study plasma-neutralized ion beam was applied to investigate ion charge effect on induction of DNA transfer. Argon ion beam at 7.5 keV was neutralized by RF-driven plasma in the beam path and then bombarded cellulose membranes which were used as the mimetic plant cell envelope. Electrical properties such as impedance and capacitance of the membranes were measured after the bombardment. An in vitro experiment on plasmid DNA transfer through the cellulose membrane was followed up. The results showed that the ion charge input played an important role in the impedance and capacitance changes which would affect DNA transfer. Generally speaking, neutral particle beam bombardment of biologic cells was more effective in inducing DNA transfer than charged ion beam bombardment.

  2. Neutralizing antibodies obtained in a persistent immune response are effective against deleterious effects induced by the Thalassophryne nattereri fish venom.

    PubMed

    Piran-Soares, Ana Amélia; Komegae, Evilin Naname; Souza, Valdênia Maria Oliveira; Fonseca, Luiz Alberto; Lima, Carla; Lopes-Ferreira, Mônica

    2007-06-01

    Thalassophryne nattereri envenoming represents a great cost to North and Northeast Brazilian communities in terms of public healths, leisure and tourism. Victims rapidally develop symptoms as pain, local swelling, erythema followed by intense necrosis that persist for long days. The aim of this work was tested the immune competence of neutralizing antibodies in pre-immunized mice against principal toxic activities induced by venom. During the primary antibody response in mice, an elevation of IgG antibody levels was only observed on day 28. After boosting, high antibody levels were detected between days 49 and 70, with a 12-fold increase in IgG level over control values at day 49. We confirmed the in vitro neutralizing capacity of T. nattereri anti-venom against toxic effects and thereafter we show that neutralizing antibodies obtained in a persistent immune response are more effective, inclusive against edematous reaction. After boosting during the secondary response mice with high antibody levels do not present any alterations in venule or arteriole after topical application of venom on cremaster muscle. In addition, CK activity diminished in these mice with high neutralizing antibody levels corroborating the attenuation of the myonecrotic effect by venom. In addition, we determined the presence of high IgG antibodies levels in patients 6 months after injury by T. nattereri. In conclusion, the presence of neutralizing antibodies against to T. nattereri venom in the serum of pre-immunized mice could change the outcome of lesion at site of posterior envenoming. Antigen-specific antibodies of high affinity in consequence to specific immune response, dependent of T lymphocyte activation, could minimize the symptoms of intense and immediate inflammatory reaction caused by T. nattereri venom. These finding prompt us to the possibility of development of immune therapeutic strategies using specific anti-venom as an efficient intervention for protecting human victims.

  3. Relativistic bounds states for a neutral particle confined to a parabolic potential induced by noninertial effects

    NASA Astrophysics Data System (ADS)

    Bakke, K.

    2010-10-01

    We obtain the solutions of the Dirac equation when the noninertial effects of the Fermi-Walker reference frame break the relativistic Landau-Aharonov-Casher quantization, but they provide bound states in an analogous way to a Dirac neutral particle subject to Tan-Inkson quantum dot potential [W.-C. Tan, J.C. Inkson, Semicond. Sci. Technol. 11 (1996) 1635].

  4. THE LINE WIDTH DIFFERENCE OF NEUTRALS AND IONS INDUCED BY MHD TURBULENCE

    SciTech Connect

    Xu, Siyao; Lazarian, A.; Yan, Huirong

    2015-09-01

    We address the problem of the different line widths of coexistent neutrals and ions observed in molecular clouds and explore whether this difference can arise from the effects of magnetohydrodynamic (MHD) turbulence acting on partially ionized gas. Among the three fundamental modes of MHD turbulence, we find that fast and slow modes do not contribute to line width differences. We focus on the Alfvénic component, and consider the damping of Alfvén modes by taking into account both neutral-ion collisions and neutral viscosity. We confirm that the line width difference can be explained by the differential damping of the Alfvénic turbulence in ions and the hydrodynamic turbulence in neutrals, and find it strongly depends on the properties of MHD turbulence. We consider various regimes of turbulence corresponding to different media magnetizations and turbulent drivings. In the case of super-Alfvénic turbulence, when the damping scale of Alfvénic turbulence is below the Alfvénic scale l{sub A}, the line width difference does not depend on magnetic field strength. In other turbulence regimes, however, the dependence is present and evaluation of magnetic field from the observed line width difference is possible.

  5. Cysteamine-induced inhibition of acid neutralization and the increase in hydrogen ion back-diffusion in duodenal mucosa

    SciTech Connect

    Ohe, K.; Okada, Y.; Fujiwara, T.; Inoue, M.; Miyoshi, A.

    1982-03-01

    To investigate the possible impairment of defensive mechanisms in cysteamine-induced duodenal ulceration, the effect of cysteamine on the neutralization of acid by the duodenum and the back-diffusion of hydrogen ions into the duodenal mucosa has been studied. The results obtained were as follows. (1) The intraduodenal pH started to decrease between 3 and 4 hr after cysteamine injection. (2) By perfusion of the duodenal loop excluding the opening of bile and pancreatic ducts, the amount of hydrogen ions (H+) neutralized was found to be significantly lower in cysteamine-treated animals than in the controls. (3) the back-diffusion of luminal H+ into the duodenal mucosa, estimated by measuring the H+ disappearance from the test solution including 100 mM HCl, was significantly increased by cysteamine. From these findings, it has been concluded that cysteamine reduces the resistance of duodenal mucosa to acid coming from the stomach.

  6. All eyes on the next generation of HIV vaccines: strategies for inducing a broadly neutralizing antibody response.

    PubMed

    Ahlers, Jeffrey D

    2014-04-01

    HIV-1 broadly neutralizing antibodies (BNAbs) develop after several years of infection through a recursive process of memory B cell adaptation and maturation against co-evolving virus quasispecies. Advances in single-cell sorting and memory B cell antibody cloning methods have identified many new HIV BNAbs targeting conserved epitopes on the HIV envelope (env) protein. 3D crystal structures and biophysical analyses of BNAbs bound to invariant virus structures expressed on monomeric gp120, epitope scaffolds, core structures, and native trimers have helped us to visualize unique binding interactions and paratope orientations that have been instrumental in guiding vaccine design. A paradigm shift in the approach to structure-based design of HIV-1 envelope immunogens came recently after several laboratories discovered that native viral envelopes or "env-structures" reverse-engineered to bind with high affinity to a handful of broadly neutralizing antibodies did not in fact bind the predicted germline precursors of these broadly neutralizing antibodies. A major challenge for HIV-1 B cell vaccine development moving forward is the design of new envelope immunogens that can trigger the selection and expansion of germline precursor and intermediate memory B cells to recapitulate B cell ontogenies associated with the maturation of a broadly neutralizing antibody response. Equally important for vaccine development is the identification of delivery systems, prime-boost strategies, and synergistic adjuvant combinations that can induce the magnitude and quality of antigen-specific T follicular helper (TFH) cell responses needed to drive somatic hypermutation (SHM) and B cell maturation against heterologous primary virus envelopes. Finding the combination of multi-protein envelope immunogens and immunization strategies that can evolve a potent broadly neutralizing antibody response portends to require a complex vaccine regimen that might be difficult to implement on any scale

  7. Neutralization of interleukin-17A delays progression of silica-induced lung inflammation and fibrosis in C57BL/6 mice.

    PubMed

    Chen, Ying; Li, Cuiying; Weng, Dong; Song, Laiyu; Tang, Wen; Dai, Wujing; Yu, Ye; Liu, Fangwei; Zhao, Ming; Lu, Chunwei; Chen, Jie

    2014-02-15

    Silica exposure can cause lung inflammation and fibrosis, known as silicosis. Interleukin-17A (IL-17A) and Th17 cells play a pivotal role in controlling inflammatory diseases. However, the roles of IL-17A and Th17 cells in the progress of silica-induced inflammation and fibrosis are poorly understood. This study explored the effects of IL-17A on silica-induced inflammation and fibrosis. We used an anti-mouse IL-17A antibody to establish an IL-17A-neutralized mice model, and mice were exposed to silica to establish an experimental silicosis model. We showed that IL-17A neutralization delayed neutrophil accumulation and progression of silica-induced lung inflammation and fibrosis. IL-17A neutralization reduced the percentage of Th17 in CD4+ T cells, decreased IL-6 and IL-1β expression, and increased Tregs at an early phase of silica-induced inflammation. Neutralization of IL-17A delayed silica-induced Th1/Th2 immune and autoimmune responses. These results suggest that IL-17A neutralization alleviates early stage silica-induced lung inflammation and delays progression of silica-induced lung inflammation and fibrosis. Neutralization of IL-17A suppressed Th17 cell development by decreasing IL-6 and/or IL-1β and increased Tregs at an early phase of silica-induced inflammation. Neutralization of IL-17A also delayed the Th1/Th2 immune response during silica-induced lung inflammation and fibrosis. IL-17A may play a pivotal role in the early phase of silica-induced inflammation and may mediate the Th immune response to influence silica-induced lung inflammation and fibrosis in mice.

  8. HYDROCEPHALUS IN THREE JUVENILE NORTH AMERICAN BLACK BEARS (URSUS AMERICANUS).

    PubMed

    Ferguson, Sylvia H; Novak, Janelle; Hecht, Silke; Craig, Linden E

    2016-06-01

    Hydrocephalus has been reported in a variety of species, including the North American black bear ( Ursus americanus ). This report describes three cases of hydrocephalus in this species from wild bears aged 3-4 mo considered retrospectively from necropsy records of one institution. Clinical signs included cortical blindness and ataxia. Primary gross findings were doming of the skull, gyri compression and flattening, and lateral ventricle dilation. Two cases had severe bilateral ventricular dilation with loss of the septum pellucidum; atrophy of the surrounding corpus callosum; and bilateral periventricular tears involving the caudate nuclei, internal capsule, and adjacent cerebrum. Histologically, the cases with periventricular tearing had severe axonal loss and degeneration, malacia, hemorrhage, and variable periventricular astrocytosis. All cases were likely congenital, given the bears' age and lack of an apparent acquired obstruction.

  9. Tetranucleotide microsatellite loci from the black bear (Ursus americanus)

    USGS Publications Warehouse

    Sanderlin, J.S.; Faircloth, B.C.; Shamblin, B.; Conroy, M.J.

    2009-01-01

    We describe primers and polymerase chain reaction conditions to amplify 21 tetranucleotide microsatellite DNA loci in black bears (Ursus americanus). We tested primers using individuals from two populations, one each in Georgia and Florida. Among individuals from Georgia (n = 29), primer pairs yielded an average of 2.9 alleles (range, one to four) and an average observed heterozygosity (HO) of 0.50 (range, 0.00 to 0.79). Among individuals from Florida (n = 19), primer pairs yielded an average of 5.7 alleles (range, one to 14) and an HO of 0.55 (range, 0.00 to 1.00). A comparison of previously developed markers with individuals from Georgia suggests that bear populations in Georgia and Florida have reduced allelic diversity relative to other populations. ?? 2008 The Authors.

  10. Numerical validation of axial plasma momentum lost to a lateral wall induced by neutral depletion

    SciTech Connect

    Takao, Yoshinori; Takahashi, Kazunori

    2015-11-15

    Momentum imparted to a lateral wall of a compact inductively coupled plasma thruster is numerically investigated for argon and xenon gases by a particle-in-cell simulation with Monte Carlo collisions (PIC-MCC). Axial plasma momentum lost to a lateral wall is clearly shown when axial depletion of the neutrals is enhanced, which is in qualitative agreement with the result in a recent experiment using a helicon plasma source [Takahashi et al., Phys. Rev. Lett. 114, 195001 (2015)]. The PIC-MCC calculations demonstrate that the neutral depletion causes an axially asymmetric profile of the plasma density and potential, leading to axial ion acceleration and the non-negligible net axial force exerted to the lateral wall in the opposite direction of the thrust.

  11. Anti-idiotypic antibodies induce neutralizing antibodies to bovine herpesvirus 1.

    PubMed Central

    Srikumaran, S; Onisk, D V; Borca, M V; Nataraj, C; Zamb, T J

    1990-01-01

    A neutralizing murine monoclonal antibody (mAb) of the IgG2a isotype (MM-113), specific for bovine herpesvirus 1 (BHV-1) glycoprotein gIV, was used to develop anti-idiotypic antibodies (anti-Id) in a calf. The bovine anti-Id were isolated from the serum of the immunized calf by affinity chromatography on an MM-113-Sepharose column, followed by repeated adsorption on a murine IgG2a column. The anti-Id thus obtained specifically reacted with MM-113, but not with isotype-matched controls. They also inhibited the binding of MM-113 to BHV-1 in a concentration-dependent manner. Mice immunized with the anti-Id produced neutralizing antibodies to BHV-1. The anti-Id bound to cells permissive to BHV-1 in a cell-binding radioimmunoassay (RIA). PMID:2165998

  12. Inducing unconscious stress: Cardiovascular activity in response to subliminal presentation of threatening and neutral words.

    PubMed

    van der Ploeg, Melanie M; Brosschot, Jos F; Verkuil, Bart; Gillie, Brandon L; Williams, DeWayne P; Koenig, Julian; Vasey, Michael W; Thayer, Julian F

    2017-10-01

    Stress-related cognitive processes may occur outside of awareness, here referred to as unconscious stress, and affect one's physiological state. Evidence supporting this idea would provide necessary clarification of the relationship between psychological stress and cardiovascular (CV) health problems. We tested the hypothesis that increases in mean arterial pressure (MAP) and total peripheral resistance (TPR) and decreases in heart rate variability (HRV) would be larger when threatening stimuli are presented outside of awareness, or subliminally, compared with neutral stimuli. Additionally, it was expected that trait worry and resting HRV, as common risk factors for CV disease, would moderate the effect. We presented a subliminal semantic priming paradigm to college students that were randomly assigned to the threat (n = 56) or neutral condition (n = 60) and assessed changes from baseline of MAP, TPR, and HRV. Level of trait worry was assessed with the Penn State Worry Questionnaire. The findings indicate that CV activity changed according to the hypothesized pattern: A higher MAP and TPR and a lower HRV in the threat condition compared with the neutral condition were found with practically meaningful effect sizes. However, these findings were only statistically significant for TPR. Furthermore, changes in CV activity were not moderated by trait worry or resting HRV. This is the first study to explicitly address the role of subliminally presented threat words on health-relevant outcome measures and suggests that unconscious stress can influence peripheral vascular resistance. © 2017 Society for Psychophysiological Research.

  13. Alendronate induces anti-migratory effects and inhibition of neutral phosphatases in UMR106 osteosarcoma cells.

    PubMed

    Molinuevo, M Silvina; Bruzzone, Liliana; Cortizo, Ana M

    2007-05-07

    Bisphosphonates are nonhydrolysable pyrophosphate analogues that prevent bone loss in several types of cancer. However, the mechanisms of anticancer action of bisphosphonates are not completely known. We have previously shown that nitrogen-containing bisphosphonates directly inhibit alkaline phosphatase of UMR106 rat osteosarcoma cells. In this study, we evaluated the effects of alendronate on the migration of UMR106 osteosarcoma using a model of multicellular cell spheroids, as well as the alendronate effect on neutral phosphatases. Alendronate significantly inhibited the migration of osteoblasts in a dose-dependent manner (10(-6)-10(-4) M). This effect was also dependent on calcium availability. The spheroid morphology and distribution of actin fibers were also affected by alendronate treatment. Alendronate dose-dependently inhibited neutral phosphatase activity in cell-free osteoblastic extracts as well as in osteoblasts in culture. Our results show that alendronate inhibits cell migration through mechanisms dependent on calcium, and that seem to involve inhibition of phosphotyrosine-neutral-phosphatases and disassembly of actin stress fibers.

  14. Rapid assessment of antibody-induced ricin neutralization by employing a novel functional cell-based assay.

    PubMed

    Gal, Yoav; Alcalay, Ron; Sabo, Tamar; Noy-Porat, Tal; Epstein, Eyal; Kronman, Chanoch; Mazor, Ohad

    2015-09-01

    Ricin is one of the most potent and lethal toxins known against which there is no available antidote. Currently, the most promising countermeasures against the toxin are based on neutralizing antibodies elicited by active vaccination or administered passively. A cell-based assay is widely applied for the primary screening and evaluation of anti-ricin antibodies, yet such assays are usually time-consuming (18-72 h). Here, we report of a novel assay to monitor ricin activity, based on HeLa cells that stably express the rapidly-degraded ubiquitin-luciferase (Ub-FL, half-life of 2 min). Ricin-induced arrest of protein synthesis could be quantified within 3 to 6h post intoxication (IC90 of 300 and 100 ng/ml, respectively). Furthermore, by stabilizing the intracellular levels of Ub-FL in the last hour of the assay, a 3-fold increase in the assay sensitivity was attained. We applied this assay to monitor the efficacy of a ricin holotoxin-based vaccine by measuring the formation of neutralizing antibodies throughout the immunization course. The potency of anti-ricin monoclonal antibodies (directed to either subunit of the toxin) could also be easily and accurately measured in this assay format. Owing to its simplicity, this assay may be implemented for high-throughput screening of ricin-neutralizing antibodies and for identification of small-molecule inhibitors of the toxin, as well as other ribosome-inactivating toxins.

  15. Kinetic and HPV infection effects on cross-type neutralizing antibody and avidity responses induced by Cervarix®

    PubMed Central

    Kemp, Troy J.; Safaeian, Mahboobeh; Hildesheim, Allan; Pan, Yuanji; Penrose, Kerri J.; Porras, Carolina; Schiller, John T.; Lowy, Douglas R.; Herrero, Rolando; Pinto, Ligia A.

    2012-01-01

    Background We previously demonstrated that Cervarix® elicits antibody responses against vaccine-related types for which clinical efficacy was demonstrated (HPV-31 and -45). Here, we evaluated the kinetics of neutralization titers and avidity of Cervarix®-induced antibodies up to 36 months of follow-up in unexposed and HPV infected women. Methods A subset of women who participated in the Cost Rica HPV-16/18 Vaccine Trial had pre- and post-vaccination sera tested for antibody responses to HPV-16, -18, -31, -45, and -58 using a pseudovirion-based neutralization assay, and HPV-16 antibody avidity using an HPV-16 L1 VLP (virus-like particle)-based ELISA developed in our laboratory. Results In uninfected women, neutralizing antibody titers did not reach significance until after the 3rd dose for HPV-31 (month 12, p=0.009) and HPV-45 (month 12, p=0.003), but then persisted up to month 36 (HPV-31, p=0.01; HPV-45, p=0.002). Individuals infected with HPV-16 or HPV-31 at enrollment developed a significantly higher median antibody response to the corresponding HPV type after one dose, but there was not a difference between median titers after three doses compared to the HPV negative group. Median HPV-16 antibody avidity and titer increased over time up to month 12; however, the HPV-16 avidity did not correlate well with HPV-16 neutralizing antibody titers at each time point examined, except for month 6. The median avidity levels were higher in HPV-16 infected women at month 1 (p=0.04) and lower in HPV-16 infected women at month 12 (p=0.006) compared to the HPV negative women. Conclusions The persistence of cross-neutralization titers at month 36 suggests cross-reactive antibody responses are likely to persist long-term and are not influenced by infection status at enrollment. However, the weak correlation between avidity and neutralization titers emphasizes the need for examining avidity in efficacy studies to determine if high avidity antibodies play a critical role in

  16. Molecular immune response of the American lobster (Homarus americanus) to the White Spot Syndrome Virus.

    PubMed

    Clark, K Fraser; Greenwood, Spencer J; Acorn, Adam R; Byrne, Philip J

    2013-11-01

    The adult American lobster (Homarus americanus) is susceptible to few naturally occurring pathogens, and no viral pathogen is known to exist. Despite this, relatively little is known about the H. americanus immune system and nothing is known about its potential viral immune response. Hundreds of rural communities in Atlantic Canada rely on the lobster fishery for their economic sustainability and could be devastated by large-scale pathogen-mediated mortality events. The White Spot Syndrome Virus is the most economically devastating viral pathogen to global shrimp aquaculture production and has been proposed to be capable of infecting all decapod crustaceans including the European Lobster. An in vivo WSSV injection challenge was conducted in H. americanus and WSSV was found to be capable of infecting and replicating within lobsters held at 20°C. The in vivo WSSV challenge also generated the first viral disease model of H. americanus and allowed for the high-throughput examination of transcriptomic changes that occur during viral infection. Microarray analysis found 136 differentially expressed genes and the expression of a subset of these genes was verified using RT-qPCR. Anti-lipopolysaccharide isoforms and acute phase serum amyloid protein A expression did not change during WSSV infection, contrary to previous findings during bacterial and parasitic infection of H. americanus. This, along with the differential gene expression of thioredoxin and trypsin isoforms, provides compelling evidence that H. americanus is capable of mounting an immune response specific to infection by different pathogen classes.

  17. Neutralizing capacity of murine sera induced by different antigens of scorpion venom.

    PubMed

    Calderon-Aranda, E S; Hozbor, D; Possani, L D

    1993-03-01

    Approximately 700 people die in Mexico each year from scorpion stings. The only useful therapy available is antiserum obtained from horses immunized with macerates of venomous gland from scorpions of the genus Centruroides. We report the results of experiments conducted with mice and rats in order to evaluate the relevant components of the venom from Centruroides noxius in the induction of a protective response against scorpion envenomation, either in vivo or in vitro. Gland macerates of whole telsons (stinger), soluble venom extracted by electrical stimulation, toxic fractions from gel filtration on Sephadex G-50 and highly purified toxin 2 from this scorpion venom were all used to produce hyperimmune mice and rats, which were challenged in vivo with the equivalent of the lethal dose 50% (LD50) of soluble venom, or their sera were prepared for in vitro neutralization experiments using non-immunized animals. The maximum neutralizing capacity (100%) was obtained when soluble venom was used as antigen, while purified toxin 2 produces 80% survival in vivo. The neutralizing capacity of murine antisera evaluated in vitro was: sera antifraction II > antitoxin 2 > antitotal venom > anti-gland macerates of whole telsons. Two additional aspects were further investigated in the present work. One is the demonstration by immunoblotting that proteins corresponding to the electrophoretic mobility of toxins known to block sodium channels are highly immunodominant in this venom. Second, there is a strong cross-reactivity of antisera produced with Centruroides noxius when assayed against venoms from other dangerous species of Centruroides scorpions from Mexico, but not against the Israeli scorpion Leiurus quinquestriatus. Finally, the immunodominance of toxic fractions in the immune response was observed either with immunization using Freund's adjuvant or by means of adsorption to nitrocellulose membranes. This latter vehicle was shown to be an excellent detoxifying agent, without

  18. Acute hepatitis C virus infection induces anti-host cell receptor antibodies with virus-neutralizing properties.

    PubMed

    Tawar, Rajiv G; Colpitts, Che C; Timm, Jörg; Fehm, Tanja; Roggendorf, Michael; Meisel, Helga; Meyer, Nicolas; Habersetzer, François; Cosset, François-Loïc; Berg, Thomas; Zeisel, Mirjam B; Baumert, Thomas F

    2015-09-01

    Hepatitis C virus (HCV) causes persistent infection in the majority of infected individuals. The mechanisms of persistence and clearance are only partially understood. Antibodies (Abs) against host cell entry receptors have been shown to inhibit HCV infection in cell culture and animal models. In this study, we aimed to investigate whether anti-receptor Abs are induced during infection in humans in vivo and whether their presence is associated with outcome of infection. We established an enzyme-linked immunosorbant assay using a recombinant CD81-claudin-1 (CLDN1) fusion protein to detect and quantify Abs directed against extracellular epitopes of the HCV CD81-CLDN1 coreceptor complex. The presence of anti-receptor Abs was studied in serum of patients from a well-defined cohort of a single-source HCV outbreak of pregnant women and several control groups, including uninfected pregnant women, patients with chronic hepatitis B and D virus (HBV/HDV) infection, and healthy individuals. Virus-neutralizing activity of Abs was determined using recombinant cell culture-derived HCV (HCVcc). Our results demonstrate that HCV-infected patients have statistically significantly higher anti-CD81/CLDN1 Ab titers during the early phase of infection than controls. The titers were significantly higher in resolvers compared to persisters. Functional studies using immunoadsorption and HCV cell culture models demonstrate that HCV-neutralizing anti-receptor Abs are induced in the early phase of HCV infection, but not in control groups. The virus-neutralizing properties of these Abs suggest a role for control of viral infection in conjunction with antiviral responses. Characterization of these anti-receptor Abs opens new avenues to prevent and treat HCV infection. © 2015 by the American Association for the Study of Liver Diseases.

  19. Opioid Exacerbation of Gram-positive sepsis, induced by Gut Microbial Modulation, is Rescued by IL-17A Neutralization

    PubMed Central

    Meng, Jingjing; Banerjee, Santanu; Li, Dan; Sindberg, Gregory M.; Wang, Fuyuan; Ma, Jing; Roy, Sabita

    2015-01-01

    Sepsis is the predominant cause of mortality in ICUs, and opioids are the preferred analgesic in this setting. However, the role of opioids in sepsis progression has not been well characterized. The present study demonstrated that morphine alone altered the gut microbiome and selectively induced the translocation of Gram-positive gut bacteria in mice. Using a murine model of poly-microbial sepsis, we further demonstrated that morphine treatment led to predominantly Gram-positive bacterial dissemination. Activation of TLR2 by disseminated Gram-positive bacteria induced sustained up-regulation of IL-17A and IL-6. We subsequently showed that overexpression of IL-17A compromised intestinal epithelial barrier function, sustained bacterial dissemination and elevated systemic inflammation. IL-17A neutralization protected barrier integrity and improved survival in morphine-treated animals. We further demonstrated that TLR2 expressed on both dendritic cells and T cells play essential roles in IL-17A production. Additionally, intestinal sections from sepsis patients on opioids exhibit similar disruption in gut epithelial integrity, thus establishing the clinical relevance of this study. This is the first study to provide a mechanistic insight into the opioid exacerbation of sepsis and show that neutralization of IL-17A might be an effective therapeutic strategy to manage Gram-positive sepsis in patients on an opioid regimen. PMID:26039416

  20. Repeated Long-Term DT Application in the DEREG Mouse Induces a Neutralizing Anti-DT Antibody Response.

    PubMed

    Wang, Junhua; Siffert, Myriam; Spiliotis, Markus; Gottstein, Bruno

    2016-01-01

    Regulatory T (Tregs) cells play an important role in mediating tolerance to self-antigens but can also mediate detrimental tolerance to tumours and pathogens in a Foxp3-dependent manner. Genetic tools exploiting the foxp3 locus including bacterial artificial chromosome- (BAC-) transgenic DEpletion of REGulatory T cells (DEREG) mice have provided essential information on Treg biology and the potential therapeutic modulation of tolerance. In DEREG mice, Foxp3(+) Tregs selectively express enhanced green fluorescent protein (eGFP) and diphtheria toxin (DT) receptor, allowing for the specific depletion of Tregs through DT administration. We here provide a detailed overview about an important consideration that long-term administration of DT induces a humoral immune response with an appropriate production of anti-DT antibodies that can inactivate DT and thus abrogate its effect in the DEREG mouse. Additionally, we showed that anti-DT mouse serum partially neutralized DT-induced Foxp3 inhibition.

  1. Repeated Long-Term DT Application in the DEREG Mouse Induces a Neutralizing Anti-DT Antibody Response

    PubMed Central

    Wang, Junhua; Siffert, Myriam; Spiliotis, Markus

    2016-01-01

    Regulatory T (Tregs) cells play an important role in mediating tolerance to self-antigens but can also mediate detrimental tolerance to tumours and pathogens in a Foxp3-dependent manner. Genetic tools exploiting the foxp3 locus including bacterial artificial chromosome- (BAC-) transgenic DEpletion of REGulatory T cells (DEREG) mice have provided essential information on Treg biology and the potential therapeutic modulation of tolerance. In DEREG mice, Foxp3+ Tregs selectively express enhanced green fluorescent protein (eGFP) and diphtheria toxin (DT) receptor, allowing for the specific depletion of Tregs through DT administration. We here provide a detailed overview about an important consideration that long-term administration of DT induces a humoral immune response with an appropriate production of anti-DT antibodies that can inactivate DT and thus abrogate its effect in the DEREG mouse. Additionally, we showed that anti-DT mouse serum partially neutralized DT-induced Foxp3 inhibition. PMID:28074191

  2. A synthetic peptide derived from domain III envelope glycoprotein of Dengue virus induces neutralizing antibody.

    PubMed

    Mary, J Asnet; Jittmittraphap, Akanitt; Chattanadee, Siriporn; Leaungwutiwong, Pornsawan; Shenbagarathai, R

    2017-09-25

    Dengue virus (DENV) is an arthropod-borne human pathogen that represents a severe public health threat in both endemic and non-endemic regions. So far, there is no licensed vaccine or specific drugs available for dengue fever. A fifteen-amino-acid-long peptide that includes the NGR motif was chemically synthesized and conjugated with keyhole limpet hemocyanin. A standard immunization protocol was followed for the production of polyclonal antibodies by immunizing rabbits against the synthetic peptide. The immune response elicited high-titer polyclonal antibodies with the reactivity of the anti-peptide antibody against both synthetic peptide and four serotypes of DENV confirmed by DOT-ELISA. Neutralizing activity of anti-peptide antibody was found to be cross-reactive and effective resulting in 60% reduction of infectivity at 1:200 dilution in all four serotypes of DENV. Our findings have the potential to further improve our understanding of virus-host interactions and provide new insights into neutralizing antibodies and could also be used as a drug target.

  3. Evidence of neutral radical induced analyte ion transformations in APPI and near-VUV APLI.

    PubMed

    Kersten, Hendrik; Funcke, Valerie; Lorenz, Matthias; Brockmann, Klaus J; Benter, Thorsten; O'Brien, Rob

    2009-10-01

    We report on the reactions of neutral radical species [OH, Cl, O(3P), H], generated in a typical atmospheric pressure ionization (API) source upon irradiation of the sample gases with either 193 nm laser radiation or 124 nm VUV light, the latter commonly used in atmospheric pressure photoionization (APPI). The present investigations focus on the polycyclic aromatic hydrocarbon pyrene as representative of the aromatic compound class. Experimental results are supported by computational methods: simple kinetic models are used to estimate the temporal evolution of the concentrations of reactants, intermediates, and final products, whereas density functional theory (DFT) energy calculations are carried out to further elucidate the proposed reaction pathways. The neutral radicals are generated upon photolysis of background water and oxygen always present in appreciable mixing ratios in typical API sources. Substantial amounts of oxygenated analyte product ions are observed using both techniques. In contrast, upon atmospheric pressure laser ionization (APLI) with 248 nm radiation, oxygenated products are virtually absent. In addition, kinetic data evaluation yielded a bimolecular rate constant of k = (1.9 +/- 0.9) x 10(-9) cm3 molecule(-1) s(-1) for the reaction of the pyrene radical cation with OH radicals.

  4. Stress-induced neutral lipid biosynthesis in microalgae - Molecular, cellular and physiological insights.

    PubMed

    Zienkiewicz, Krzysztof; Du, Zhi-Yan; Ma, Wei; Vollheyde, Katharina; Benning, Christoph

    2016-09-01

    Photosynthetic microalgae have promise as biofuel feedstock. Under certain conditions, they produce substantial amounts of neutral lipids, mainly in the form of triacylglycerols (TAGs), which can be converted to fuels. Much of our current knowledge on the genetic and molecular basis of algal neutral lipid metabolism derives mainly from studies of plants, i.e. seed tissues, and to a lesser extent from direct studies of algal lipid metabolism. Thus, the knowledge of TAG synthesis and the cellular trafficking of TAG precursors in algal cells is to a large extent based on genome predictions, and most aspects of TAG metabolism have yet to be experimentally verified. The biofuel prospects of microalgae have raised the interest in mechanistic studies of algal TAG biosynthesis in recent years and resulted in an increasing number of publications on lipid metabolism in microalgae. In this review we summarize the current findings on genetic, molecular and physiological studies of TAG accumulation in microalgae. Special emphasis is on the functional analysis of key genes involved in TAG synthesis, molecular mechanisms of regulation of TAG biosynthesis, as well as on possible mechanisms of lipid droplet formation in microalgal cells. This article is part of a Special Issue entitled: Plant Lipid Biology edited by Kent D. Chapman and Ivo Feussner.

  5. Analysis of gene expression in Homarus americanus larvae exposed to sublethal concentrations of endosulfan during metamorphosis.

    PubMed

    Bauer, Megan; Greenwood, Spencer J; Clark, K Fraser; Jackman, Paula; Fairchild, Wayne

    2013-12-01

    Agricultural pesticide runoff has been suspected as the cause of numerous fish kills in rivers throughout Prince Edward Island but the impact on the surrounding marine environment is unknown. Endosulfan, an organochlorine pesticide, is a potent neurotoxin and molt inhibitor used to combat the Colorado potato beetle however it has the potential to affect non-target organisms including the American lobster (Homarus americanus). Metamorphosis is a critical stage of development and the effects of contaminant exposure during this time are largely unknown in lobster. A 14day endosulfan exposure was performed to identify the effects on survival, development and gene expression in lobster larvae during metamorphosis; all of which were predicted to be negatively impacted. The higher endosulfan concentrations resulted in greater mortality and a significant increase in the number of days required to reach metamorphosis in surviving animals. A custom made H. americanus microarray was used for monitoring the changes in expression of 14,592 genes at the termination of the exposure. Genes with >1.5 fold change and identified as being significant at p<0.05 using one-way ANOVA were selected for further analysis. A total of 707 genes were identified as being significantly differentiated, however with only ~40% annotation of the array, the majority of these genes were unknown. Annotated genes of interest were involved in many processes: development, metabolism, immunity and oxidative stress response and gene regulation. Nine genes of interest (histone H1, farnesoic acid O-methyltransferase, cuticle protein, glutathione S-transferase, thioredoxin, NADH dehydrogenase, ecdysone nuclear receptor Fushi tarazu F1 (FTZ-F1), ferritin and ecdysone inducible protein E75 (EIP-E75)) were selected for RT-qPCR validation of the microarray results. The RT-qPCR method was more sensitive than the microarray yet detected similar expression patterns. The two highest endosulfan concentrations resulted

  6. Single photon production induced by (anti)neutrino neutral current scattering on nucleons and nuclear targets

    SciTech Connect

    Alvarez-Ruso, L.; Nieves, J.; Wang, E.

    2015-10-15

    We review our theoretical approach to neutral current photon emission on nucleons and nuclei in the few-GeV energy region, relevant for neutrino oscillation experiments. These reactions are dominated by the weak excitation of the Δ(1232) resonance but there are also important non-resonant contributions. We have also included terms mediated by nucleon excitations from the second resonance region. On nuclei, Pauli blocking, Fermi motion and the in-medium Δ resonance broadening have been taken into account for both incoherent and coherent reaction channels. With this model, the number and distributions of photon events at the MiniBooNE and T2K experiments have been obtained. We have also compared to the NOMAD upper limit at higher energies. The implications of our findings and future perspectives are discussed.

  7. Recombinant outer membrane vesicles carrying Chlamydia muridarum HtrA induce antibodies that neutralize chlamydial infection in vitro.

    PubMed

    Bartolini, Erika; Ianni, Elvira; Frigimelica, Elisabetta; Petracca, Roberto; Galli, Giuliano; Berlanda Scorza, Francesco; Norais, Nathalie; Laera, Donatello; Giusti, Fabiola; Pierleoni, Andrea; Donati, Manuela; Cevenini, Roberto; Finco, Oretta; Grandi, Guido; Grifantini, Renata

    2013-01-01

    Outer membrane vesicles (OMVs) are spheroid particles released by all Gram-negative bacteria as a result of the budding out of the outer membrane. Since they carry many of the bacterial surface-associated proteins and feature a potent built-in adjuvanticity, OMVs are being utilized as vaccines, some of which commercially available. Recently, methods for manipulating the protein content of OMVs have been proposed, thus making OMVs a promising platform for recombinant, multivalent vaccines development. Chlamydia muridarum DO serine protease HtrA, an antigen which stimulates strong humoral and cellular responses in mice and humans, was expressed in Escherichia coli fused to the OmpA leader sequence to deliver it to the OMV compartment. Purified OMVs carrying HtrA (CM rHtrA-OMV) were analyzed for their capacity to induce antibodies capable of neutralizing Chlamydia infection of LLC-MK2 cells in vitro. CM rHtrA-OMV immunization in mice induced antibodies that neutralize Chlamydial invasion as judged by an in vitro infectivity assay. This was remarkably different from what observed with an enzymatically functional recombinant HtrA expressed in, and purified from the E. coli cytoplasm (CM rHtrA). The difference in functionality between anti-CM rHtrA and anti-CM rHtrA-OMV antibodies was associated to a different pattern of protein epitopes recognition. The epitope recognition profile of anti-CM HtrA-OMV antibodies was similar to that induced in mice during Chlamydial infection. When expressed in OMVs HtrA appears to assume a conformation similar to the native one and this results in the elicitation of functional immune responses. These data further support the potentiality of OMVs as vaccine platform.

  8. Recombinant outer membrane vesicles carrying Chlamydia muridarum HtrA induce antibodies that neutralize chlamydial infection in vitro

    PubMed Central

    Bartolini, Erika; Ianni, Elvira; Frigimelica, Elisabetta; Petracca, Roberto; Galli, Giuliano; Berlanda Scorza, Francesco; Norais, Nathalie; Laera, Donatello; Giusti, Fabiola; Pierleoni, Andrea; Donati, Manuela; Cevenini, Roberto; Finco, Oretta; Grandi, Guido; Grifantini, Renata

    2013-01-01

    Background Outer membrane vesicles (OMVs) are spheroid particles released by all Gram-negative bacteria as a result of the budding out of the outer membrane. Since they carry many of the bacterial surface-associated proteins and feature a potent built-in adjuvanticity, OMVs are being utilized as vaccines, some of which commercially available. Recently, methods for manipulating the protein content of OMVs have been proposed, thus making OMVs a promising platform for recombinant, multivalent vaccines development. Methods Chlamydia muridarum DO serine protease HtrA, an antigen which stimulates strong humoral and cellular responses in mice and humans, was expressed in Escherichia coli fused to the OmpA leader sequence to deliver it to the OMV compartment. Purified OMVs carrying HtrA (CM rHtrA-OMV) were analyzed for their capacity to induce antibodies capable of neutralizing Chlamydia infection of LLC-MK2 cells in vitro. Results CM rHtrA-OMV immunization in mice induced antibodies that neutralize Chlamydial invasion as judged by an in vitro infectivity assay. This was remarkably different from what observed with an enzymatically functional recombinant HtrA expressed in, and purified from the E. coli cytoplasm (CM rHtrA). The difference in functionality between anti-CM rHtrA and anti-CM rHtrA-OMV antibodies was associated to a different pattern of protein epitopes recognition. The epitope recognition profile of anti-CM HtrA-OMV antibodies was similar to that induced in mice during Chlamydial infection. Conclusions When expressed in OMVs HtrA appears to assume a conformation similar to the native one and this results in the elicitation of functional immune responses. These data further support the potentiality of OMVs as vaccine platform. PMID:24009891

  9. Elevated expression of protein biosynthesis genes in liver and muscle of hibernating black bears (Ursus americanus).

    PubMed

    Fedorov, Vadim B; Goropashnaya, Anna V; Tøien, Oivind; Stewart, Nathan C; Gracey, Andrew Y; Chang, Celia; Qin, Shizhen; Pertea, Geo; Quackenbush, John; Showe, Louise C; Showe, Michael K; Boyer, Bert B; Barnes, Brian M

    2009-04-10

    We conducted a large-scale gene expression screen using the 3,200 cDNA probe microarray developed specifically for Ursus americanus to detect expression differences in liver and skeletal muscle that occur during winter hibernation compared with animals sampled during summer. The expression of 12 genes, including RNA binding protein motif 3 (Rbm3), that are mostly involved in protein biosynthesis, was induced during hibernation in both liver and muscle. The Gene Ontology and Gene Set Enrichment analysis consistently showed a highly significant enrichment of the protein biosynthesis category by overexpressed genes in both liver and skeletal muscle during hibernation. Coordinated induction in transcriptional level of genes involved in protein biosynthesis is a distinctive feature of the transcriptome in hibernating black bears. This finding implies induction of translation and suggests an adaptive mechanism that contributes to a unique ability to reduce muscle atrophy over prolonged periods of immobility during hibernation. Comparing expression profiles in bears to small mammalian hibernators shows a general trend during hibernation of transcriptional changes that include induction of genes involved in lipid metabolism and carbohydrate synthesis as well as depression of genes involved in the urea cycle and detoxification function in liver.

  10. Lipoproteins/peptides are sepsis-inducing toxins from bacteria that can be neutralized by synthetic anti-endotoxin peptides

    PubMed Central

    de Tejada, Guillermo Martinez; Heinbockel, Lena; Ferrer-Espada, Raquel; Heine, Holger; Alexander, Christian; Bárcena-Varela, Sergio; Goldmann, Torsten; Correa, Wilmar; Wiesmüller, Karl-Heinz; Gisch, Nicolas; Sánchez-Gómez, Susana; Fukuoka, Satoshi; Schürholz, Tobias; Gutsmann, Thomas; Brandenburg, Klaus

    2015-01-01

    Sepsis, a life-threatening syndrome with increasing incidence worldwide, is triggered by an overwhelming inflammation induced by microbial toxins released into the bloodstream during infection. A well-known sepsis-inducing factor is the membrane constituent of Gram-negative bacteria, lipopolysaccharide (LPS), signalling via Toll-like receptor-4. Although sepsis is caused in more than 50% cases by Gram-positive and mycoplasma cells, the causative compounds are still poorly described. In contradicting investigations lipoproteins/-peptides (LP), lipoteichoic acids (LTA), and peptidoglycans (PGN), were made responsible for eliciting this pathology. Here, we used human mononuclear cells from healthy donors to determine the cytokine-inducing activity of various LPs from different bacterial origin, synthetic and natural, and compared their activity with that of natural LTA and PGN. We demonstrate that LP are the most potent non-LPS pro-inflammatory toxins of the bacterial cell walls, signalling via Toll-like receptor-2, not only in vitro, but also when inoculated into mice: A synthetic LP caused sepsis-related pathological symptoms in a dose-response manner. Additionally, these mice produced pro-inflammatory cytokines characteristic of a septic reaction. Importantly, the recently designed polypeptide Aspidasept® which has been proven to efficiently neutralize LPS in vivo, inhibited cytokines induced by the various non-LPS compounds protecting animals from the pro-inflammatory activity of synthetic LP. PMID:26390973

  11. Lipoproteins/peptides are sepsis-inducing toxins from bacteria that can be neutralized by synthetic anti-endotoxin peptides.

    PubMed

    Martinez de Tejada, Guillermo; Heinbockel, Lena; Ferrer-Espada, Raquel; Heine, Holger; Alexander, Christian; Bárcena-Varela, Sergio; Goldmann, Torsten; Correa, Wilmar; Wiesmüller, Karl-Heinz; Gisch, Nicolas; Sánchez-Gómez, Susana; Fukuoka, Satoshi; Schürholz, Tobias; Gutsmann, Thomas; Brandenburg, Klaus

    2015-09-22

    Sepsis, a life-threatening syndrome with increasing incidence worldwide, is triggered by an overwhelming inflammation induced by microbial toxins released into the bloodstream during infection. A well-known sepsis-inducing factor is the membrane constituent of Gram-negative bacteria, lipopolysaccharide (LPS), signalling via Toll-like receptor-4. Although sepsis is caused in more than 50% cases by Gram-positive and mycoplasma cells, the causative compounds are still poorly described. In contradicting investigations lipoproteins/-peptides (LP), lipoteichoic acids (LTA), and peptidoglycans (PGN), were made responsible for eliciting this pathology. Here, we used human mononuclear cells from healthy donors to determine the cytokine-inducing activity of various LPs from different bacterial origin, synthetic and natural, and compared their activity with that of natural LTA and PGN. We demonstrate that LP are the most potent non-LPS pro-inflammatory toxins of the bacterial cell walls, signalling via Toll-like receptor-2, not only in vitro, but also when inoculated into mice: A synthetic LP caused sepsis-related pathological symptoms in a dose-response manner. Additionally, these mice produced pro-inflammatory cytokines characteristic of a septic reaction. Importantly, the recently designed polypeptide Aspidasept(®) which has been proven to efficiently neutralize LPS in vivo, inhibited cytokines induced by the various non-LPS compounds protecting animals from the pro-inflammatory activity of synthetic LP.

  12. Time-resolved imaging of laser-induced vibrational wave packets in neutral and ionic states of iodomethane

    NASA Astrophysics Data System (ADS)

    Malakar, Y.; Kaderiya, B.; Zohrabi, M.; Pearson, W. L.; Ziaee, F.; Kananka Raju, P.; Ben-Itzhak, I.; Rolles, D.; Rudenko, A.

    2016-05-01

    Light-driven vibrational wave packets play an important role in molecular imaging and coherent control applications. Here we present the results of a pump-probe experiment characterizing laser-induced vibrational wave packets in both, neutral and ionic states of CH3 I (iodomethane), one of the prototypical polyatomic systems. Measuring yields and kinetic energies of all ionic fragments as a function of the time delay between two 25 fs, 800 nm pump and probe pulses, we map vibrational motion of the molecule, and identify the states involved by channel-resolved Fourier spectroscopy. In the Coulomb explosion channels we observe features with ~ 130 fs periodicity resulting from C-I symmetric stretch (ν3 mode) of the electronically excited cationic state. However the Fourier transform of the low-energy I+ ion yield produced by the dissociative ionization of CH3 I reveals the signatures of the same vibrational mode in the ground electronic states of both, neutral and cation, reflected in 65-70 fs oscillations. We observe the degeneration of the oscillatory structures from the cationic states within ~ 2 ps and discuss most likely reasons for this behavior. Supported by the Chemical Sciences, Geosciences, and Biosciences Division, Office of Basic Energy Sciences, Office of Science, U. S. DOE. K. R. P. and W. L. P. supported by NSF Award No. IIA-143049.

  13. Transcutaneous delivery of tetanus toxin Hc fragment induces superior tetanus toxin neutralizing antibody response compared to tetanus toxoid.

    PubMed

    Johnston, Louise; Mawas, Fatme; Tierney, Rob; Qazi, Omar; Fairweather, Neil; Sesardic, Dorothea

    2009-04-01

    Transcutaneous immunization is a promising vaccination delivery strategy which targets potent immune cells residing in the outer layer of the skin. In this study, the immunogenicity and neutralizing potency of the non-toxic Hc fragment of tetanus toxin (HcWT) and a mutant of Hc lacking ganglioside binding activity were compared with that of tetanus toxoid (TTxd) following transcutaneous immunization (TCI) of mice. Mice immunized with HcWT in the absence of an adjuvant induced highest anti-toxoid and anti-Hc antibody titres, with a significant increase in the toxin neutralizing antibody response compared with TTxd. These results are in contrast to previous studies employing subcutaneous delivery, where TTxd was found to be a more potent immunogen than the Hc fragment of the toxin. We conclude that the HcWT protein is more immunogenic than TTxd when given via the transcutaneous route. Our results suggest that TCI may provide an opportunity for effective delivery of toxin-like antigens which harbor protective epitopes and that traditional toxoid proteins may not be optimal antigens for skin immunization.

  14. Spatial memory in captive American black bears (Ursus americanus).

    PubMed

    Zamisch, Valeria; Vonk, Jennifer

    2012-11-01

    The spatial memory and foraging strategies of four adult captive-born American black bears (Ursus americanus) were explored in four experiments using a simulated foraging task. In the first three experiments, each session consisted of two phases separated by a delay: During the exploration phase, subjects foraged among a set of baited and unbaited sites. During the delay, the same locations were rebaited and subjects were released again and allowed to search the sites (search phase). In Experiments 1a and 1b, different sites were baited each day and the interval between exploration and search was short (4 hr or 15 min). Subjects were not accurate at recovering the food items in either experiment. In Experiment 2, an "informed forager" paradigm was used in which one subject was given privileged knowledge about the location of the food during the exploration phase and was later released with an "uninformed" competitor during the search phase. The bears did not achieve above-chance recovery accuracy even in the presence of a competitor. In Experiment 3, the same two of four sites were continually baited and the bears were released simultaneously over a period of 20 days, with each baiting separated by 2 or 3 days. As a group, the bears' foraging accuracy with repeated baiting and longer intervals approached greater than chance accuracy. Results suggest some limitations on bears' use of spatial memory in captive environments, but reveal the potential for use of spatial memory over longer delays.

  15. Subcuticular urate accumulation in an American lobster (Homarus americanus).

    PubMed

    Battison, A L

    2013-05-01

    An unusually "lumpy" lobster, Homarus americanus, was presented to the Atlantic Veterinary College Lobster Science Centre for evaluation. The lobster was weak with numerous pale, raised, and flat areas (diameter, 3-15 mm) on the exoskeleton, some of which were ulcerated. On postmortem examination, the pale areas corresponded to accumulations of viscous to free-flowing white material, which was found in only the subcuticular connective tissues. No internal organs were affected. Direct light examination of nonstained impression smears of the material showed abundant crystals resembling uric acid, amorphous urates, and sodium urate, which were readily soluble in 1 M potassium hydroxide. Wright-Giemsa stained imprints showed numerous fine, rounded, nonstaining granules free in the background and within individual round cells. Fourier-transformed infrared spectroscopy confirmed the presence of urates or mixed urate salts. Hemolymph plasma urea (1.7 mmol/liter) and uric acid (287 μmol/liter) concentrations were slightly higher than those seen with 36-hour emersion. Histologic sections showed aggregates of vacuolated mononuclear cells in the loose subcuticular connective tissue occasionally infiltrating between underlying muscle fibers. Grossly visible urate deposits are occasionally documented in land crabs and rarely reported in the blue crab; none, however, are associated with deformation of the cuticle. Possible etiologies include increased uric acid intake or production or decreased excretion. Anecdotal reports of similarly affected lobsters have been received but are intermittent and undocumented.

  16. Marburg virus-like particles produced in insect cells induce neutralizing antibodies in rhesus macaques.

    PubMed

    Gai, Weiwei; Zheng, Xuexing; Wang, Chong; Zhao, Yongkun; Wang, Qi; Wang, Hualei; Wong, Gary; Xie, Ying; Wang, Haijun; Cao, Zengguo; Feng, Na; Chi, Hang; Wang, Tiecheng; Gao, Yuwei; Shan, Junjie; Yang, Songtao; Xia, Xianzhu

    2017-04-12

    Marburg virus (MARV), which is one of the most virulent agents in the world, causes lethal haemorrhagic fever in humans and nonhuman primates (NHPs) with a mortality rate of up to 90%. Currently, there is no effective treatment or approved vaccine for MARV for human use to control disease outbreak and spread. Virus-like particles (VLPs), which are morphologically identical to the native infectious virus particle, are efficacious as vaccines against many viruses, including human papilloma virus (HPV), porcine circovirus (PCV) type 2 and hepatitis B virus (HBV). In this study, we generated MARV virus-like particles (VLPs) by co-expressing a glycoprotein (GP) and matrix protein (VP40) using the baculovirus expression system. Rhesus macaques vaccinated with MARV VLPs mixed with adjuvant Poria cocos polysaccharides (PCP-II) produced a GP-specific IgG titer of up to 1:1280 and virus-neutralizing antibody titers that reached 1:320. MARV VLPs also elicited interferon-γ (IFN-γ) and interleukin-4 (IL-4) secretion associated with T-helper 1 cell (Th1)- and T-helper 2 cell (Th2)-mediated immunity, as detected using enzyme-linked immunospot (ELISpot) assays. These data indicate that MARV VLPs mixed with adjuvant PCP-II have excellent immunogenicity in rhesus macaques and may be a promising candidate vaccine against MARV.This article is protected by copyright. All rights reserved.

  17. Analysis of Potential Glove-Induced Hand Injury Metrics During Typical Neutral Buoyancy Training Operations

    NASA Technical Reports Server (NTRS)

    McFarland, Shane

    2016-01-01

    Injuries to the hands are common among astronauts who train for extravehicular activity. When the gloves are pressurized, they restrict movement and create pressure points during tasks, sometimes resulting in pain, muscle fatigue, abrasions, and occasionally more severe injuries such as onycholysis. A brief review of NASA's Lifetime Surveillance of Astronaut Health's injury database reveals that 76 percent of astronaut hand and arm injuries from training between 1993 and 2010 occurred either to the fingernail, finger crotch, metacarpophalangeal joint, or fingertip. The purpose of this study was to assess the potential of using small sensors to measure forces acting on the fingers and hand within pressurized gloves and other variables such as skin temperature, humidity, and fingernail strain of a NASA crewmember during typical NBL (Neutral Buoyancy Laboratory) training activity. During the 5-hour exercise, the crewmember seemed to exhibit very large forces on some fingers, resulting in higher strain than seen in previous glove-box testing. In addition, vital information was collected on the glove cavity environment with respect to temperature and humidity. All of this information gathered during testing will be carried forward into future testing, potentially in glove-box- or 1G- (1 gravitational force) or NBL-suited environments, to better characterize and understand the possible causes of hand injury amongst NASA crew.

  18. Spatially induced nestedness in a neutral model of phage-bacteria networks.

    PubMed

    Valverde, Sergi; Elena, Santiago F; Solé, Ricard

    2017-07-01

    Ecological networks, both displaying mutualistic or antagonistic interactions, seem to share common structural traits: the presence of nestedness and modularity. A variety of model approaches and hypothesis have been formulated concerning the significance and implications of these properties. In phage-bacteria bipartite infection networks, nestedness seems to be the rule in many different contexts. Modeling the coevolution of a diverse virus-host ensemble is a difficult task, given the dimensionality and multi parametric nature of a standard continuous approximation. Here, we take a different approach, by using a neutral, toy model of host-phage interactions on a spatial lattice. Each individual is represented by a bit string (a digital genome) but all strings in each class (i.e. hosts or phages) share the same sets of parameters. A matching allele model of phage-virus recognition rule is enough to generate a complex, diverse ecosystem with heterogeneous patterns of interaction and nestedness, provided that interactions take place under a spatially constrained setting. It is found that nestedness seems to be an emergent property of the co-evolutionary dynamics. Our results indicate that the enhanced diversity resulting from localized interactions strongly promotes the presence of nested infection matrices.

  19. The complete genome sequence of 'Candidatus Liberibacter americanus', associated with Citrus huanglongbing.

    PubMed

    Wulff, Nelson A; Zhang, Shujian; Setubal, João C; Almeida, Nalvo F; Martins, Elaine C; Harakava, Ricardo; Kumar, Dibyendu; Rangel, Luiz Thiberio; Foissac, Xavier; Bové, Joseph M; Gabriel, Dean W

    2014-02-01

    Liberibacter spp. form a Rhizobiaceae clade of phloem-limited pathogens of limited host range. Two obligately parasitic species have been sequenced: 'Candidatus Liberibacter asiaticus', which causes citrus huanglongbing (HLB) worldwide, and 'Ca. L. solanacearum', which causes potato "zebra chip" disease. A third (proposed) species, Liberibacter crescens, was isolated from mountain papaya, grown in axenic culture, and sequenced. In an effort to identify common host determinants, the complete genomic DNA sequence of a second HLB species, 'Ca. L. americanus' strain 'São Paulo' was determined. The circular genome of 1,195,201 bp had an average 31.12% GC content and 983 predicted protein encoding genes, 800 (81.4%) of which had a predicted function. There were 658 genes common to all sequenced Liberibacter spp. and only 8 genes common to 'Ca. L. americanus' and 'Ca. L. asiaticus' but not found in 'Ca. L. solanacearum'. Surprisingly, most of the lipopolysaccharide biosynthetic genes were missing from the 'Ca. L. americanus' genome, as well as OmpA and a key regulator of flagellin, all indicating a 'Ca. L. americanus' strategy of avoiding production of major pathogen-associated molecular patterns present in 'Ca. L. asiaticus' and 'Ca. L. solanacearum'. As with 'Ca. L. asiaticus', one of two 'Ca. L. americanus' prophages replicated as an excision plasmid and carried potential lysogenic conversion genes that appeared fragmentary or degenerated in 'Ca. L. solanacearum'.

  20. In vivo neutralization of eosinophil-derived major basic protein inhibits antigen-induced bronchial hyperreactivity in sensitized guinea pigs.

    PubMed Central

    Lefort, J; Nahori, M A; Ruffie, C; Vargaftig, B B; Pretolani, M

    1996-01-01

    This study examines the effect of purified rabbit antiguinea pig eosinophil-derived major basic protein (MBP) Ig on antigen-induced bronchial hyperreactivity to inhaled acetylcholine in aerosol-sensitized guinea pigs. Ovalbumin inhalation by sensitized guinea pigs induced a rise in the numbers of eosinophils and in the levels of MBP in the bronchoalveolar lavage fluid, which peaked at 24 h and resolved at 72 h. Antigen-challenged animals exhibited bronchial hyperreactivity to inhale acetylcholine at 72 h, but not at 6 or 24 h. The intranasal administration of 200 microliter of purified rabbit anti-guinea pig MBP Ig, at 2.5 mg/ml, but not of the control preimmune rabbit Ig, 1 h before and 5 h after ovalbumin inhalation suppressed bronchial hyperreactivity to acetylcholine at 72 h without affecting the number of eosinophils accumulating in the bronchoalveolar lavage fluid. These findings indicate that antigen challenge in sensitized guinea pigs is followed by early eosinophil infiltration and activation within the airways and by late bronchial hyperreactivity. Neutralization of endogenously secreted MBP by a specific antiserum prevented antigen-induced bronchial hyperreactivity, suggesting that eosinophil degranulation plays an important role in the alterations of bronchopulmonary function in the guinea pig. PMID:8613536

  1. Lethal iron deprivation induced by non-neutralizing antibodies targeting transferrin receptor 1 in malignant B cells.

    PubMed

    Rodríguez, José A; Luria-Pérez, Rosendo; López-Valdés, Héctor E; Casero, David; Daniels, Tracy R; Patel, Shabnum; Avila, David; Leuchter, Richard; So, Sokuntheavy; Ortiz-Sánchez, Elizabeth; Bonavida, Benjamin; Martínez-Maza, Otoniel; Charles, Andrew C; Pellegrini, Matteo; Helguera, Gustavo; Penichet, Manuel L

    2011-11-01

    A number of antibodies have been developed that induce lethal iron deprivation (LID) by targeting the transferrin receptor 1 (TfR1/CD71) and either neutralizing transferrin (Tf) binding, blocking internalization of the receptor and/or inducing its degradation. We have developed recombinant antibodies targeting human TfR1 (ch128.1 and ch128.1Av), which induce receptor degradation and are cytotoxic to certain malignant B-cells. We now show that internalization of TfR1 bound to these antibodies can lead to its sequestration and degradation, as well as reduced Tf uptake, and the induction of a transcriptional response consistent with iron deprivation, which is mediated in part by downstream targets of p53. Cells resistant to these antibodies do not sequester and degrade TfR1 after internalization of the antibody/receptor complex, and accordingly maintain their ability to internalize Tf. These findings are expected to facilitate the rational design and clinical use of therapeutic agents targeting iron import via TfR1 in hematopoietic malignancies.

  2. Lethal iron deprivation induced by non-neutralizing antibodies targeting transferrin receptor 1 in malignant B cells

    PubMed Central

    Rodríguez, JoséA.; Luria-Pérez, Rosendo; López-Valdés, Héctor E.; Casero, David; Daniels, Tracy R.; Patel, Shabnum; Avila, David; Leuchter, Richard; So, Sokuntheavy; ánchez, Elizabeth Ortiz-S; Bonavida, Benjamin; Martínez-Maza, Otoniel; Charles, Andrew .C; Pellegrini, Matteo; Helguera, Gustavo; Penichet, Manuel L.

    2013-01-01

    A number of antibodies have been developed that induce lethal iron deprivation (LID) by targeting the transferrin receptor 1 (TfR1/CD71) and either neutralizing transferrin (Tf) binding, blocking internalization of the receptor and/or inducing its degradation. We have developed recombinant antibodies targeting human TfR1 (ch128.1 and ch128.1Av), which induce receptor degradation and are cytotoxic to certain malignant B-cells. We now show that internalization of TfR1 bound to these antibodies can lead to its sequestration and degradation, as well as reduced Tf uptake, and the induction of a transcriptional response consistent with iron deprivation, which is mediated in part by downstream targets of p53. Cells resistant to these antibodies do not sequester and degrade TfR1 after internalization of the antibody/receptor complex, and accordingly maintain their ability to internalize Tf. These findings are expected to facilitate the rational design and clinical use of therapeutic agents targeting iron import via TfR1 in hematopoietic malignancies. PMID:21870996

  3. A murine cytomegalovirus-neutralizing monoclonal antibody exhibits autoreactivity and induces tissue damage in vivo.

    PubMed Central

    Chapman, A J; Farrell, H E; Thomas, J A; Papadimitriou, J M; Garlepp, M J; Scalzo, A A; Shellam, G R

    1994-01-01

    The autoreactivity of murine cytomegalovirus (MCMV)-neutralizing monoclonal antibody (mAb) AC1 was examined in vitro and in vivo. Both mAb AC1 and a human antiserum reactive with U1-small nuclear ribonucleoprotein (U1-snRNP) stained uninfected mouse embryo fibroblasts (MEF) in a speckled nuclear pattern and reacted with 70,000 molecular weight (MW) MEF nuclear antigens by immunoblotting, suggesting that mAb AC1 cross-reacted with the 70,000 MW component of U1-snRNP. However, only mAb AC1 cross-reacted with an additional epithelial cytoplasmic autoantigen present in cultured HEp2 cells. On tissue sections from uninfected mice, mAb AC1 predominantly reacted with a component of central and peripheral nervous systems, although cross-reactivity with the stratum spinosum of the skin and the outer sheath of hair follicles was also observed. Immunoblotting revealed that mAb AC1 reacted with phosphorylated epitopes present on a 98,000 MW MCMV structural protein and the 200,000 MW mouse neurofilament protein (NFP). Treatment of uninfected mice with mAb AC1 resulted in a severe interstitial pneumonia with greatly thickened and congested alveolar septa. Severe oedema of the hypodermis and a mild mesangial proliferative glomerulonephritis were also observed. These results demonstrate that a mAb reacting with a MCMV structural phosphoprotein which can protect mice against the dissemination of MCMV, can also promote the development of autoimmune disease. Therefore, the production of such cross-reactive antibodies may be an important mechanism in the development of autoimmunity following viral infection. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 PMID:7515848

  4. The effect of fallow on Longidorus americanus, a nematode associated with stunting of loblolly pine seedlings in Georgia, USA

    Treesearch

    Stephen W. Fraedrich; Michelle M. Cram; Stanley J. Zarnoch

    2005-01-01

    Stunting of loblolly pine (Pinue taeda L.) seedlings, caused by Longidorus americanus, has been a problem at a Georgia (USA) nursery. Field and growth chamber studies were conducted to determine the survivability of the nematode in a fallow nursery soil. The population density of L. americanus decreased rapidly...

  5. Coagulation of charged microparticles in neutral gas and charge-induced gel transitions.

    PubMed

    Ivlev, A V; Morfill, G E; Konopka, U

    2002-11-04

    Coagulation of charged particles was studied using the mean-field Smoluchowski equation. The coagulation equation was generalized for the case of a conserved system of charged particles. It was shown that runaway cluster growth (gelation) solutions exist if the charge-dipole (induced) interaction of clusters is included. When clusters are in thermal equilibrium with the ambient gas, the charge-dipole interaction dramatically enhances the aggregation process and considerably increases the likelihood of a gelation transition.

  6. Development of a 2-Plex Luminex-Based Competitive Immunoassay to Quantify Neutralizing Antibodies Induced by Virus-Like Particles for Human Papillomavirus 16 and 18

    PubMed Central

    Lv, Pin; Zhu, Hongmei; Wang, Hongfang; Wang, George

    2011-01-01

    Human papillomavirus (HPV) L1 virus-like particles (VLPs) were proven an effective vaccine candidate to prevent against HPV-16 and -18 infections. In order to evaluate the potency of our produced HPV-16 and -18 L1 VLPs-based vaccine candidates, also to quantify neutralizing antibodies induced by them, a 2-plex Luminex-based competitive immunoassay was developed. Unlike the published paper, the no-biotin conjugated neutralizing mAbs spiked normal human serum (NHS) was used for standard curve preparation, while phycoerythrin (PE) was not labeled directly to neutralizing mAbs for signaling. After the coupling optimization of VLPs to microspheres and the neutralizing mAbs biotinylation, the 2-plex standard curve was prepared with good fit and high dynamic range. In addition, no cross-reactivity was also confirmed. The 2-plex Luminex-based immunoassay represents good potential not only for vaccine candidate's evaluation but also for its further clinical use. PMID:21808597

  7. Atrial fibrillation down-regulates renal neutral endopeptidase expression and induces profibrotic pathways in the kidney.

    PubMed

    Bukowska, Alicja; Lendeckel, Uwe; Krohn, Alexander; Keilhoff, Gerburg; ten Have, Sara; Neumann, Klaus Hinrich; Goette, Andreas

    2008-10-01

    Recent studies suggest that atrial fibrillation (AF) substantially influences microvascular flow in ventricular myocardium. This process may contribute to the occurrence of heart failure in AF. In general, development of heart failure and renal dysfunction go hand-in-hand causing systemic fluid overload and oedema. So far, it is unknown whether AF itself influences renal function. The aim of the present study was to determine the impact of AF on renal gene expression in a closed chest rapid atrial pacing model. A total of 14 pigs were studied. In five pigs, rapid atrial pacing (AT) was performed for 7 h (600 bpm); in five additional animals, rapid atrial pacing was performed in the presence of irbesartan infusion (irbesartan group). Four pigs were instrumented without interventions (sham). After the pacing period, renal expression of collagen I alpha 1 and I alpha 3, transforming growth factor-beta (TGF-beta), neutral endopeptidase (NEP; the main enzyme involved in natriuretic protein metabolism), and atrial natriuretic peptide (ANP) were determined by RT-PCR and immunoblot analysis. Functional in vitro experiments were performed using HEK-293 kidney cells. Renal mRNA expression of NEP was substantially down-regulated during AT (AT: 12.7 +/- 9.3% vs. sham: 100 +/- 43.4%; P < 0.01). Results at the mRNA level were confirmed at the protein level. Irbesartan therapy did not prevent down-regulation of NEP. In contrast, TGF-beta1 mRNA expression was up-regulated (AT: 208.5 +/- 79.3% vs. sham: 100 +/- 34.6% P< 0.05). Collagen and angiotensin II type 1 receptor (AT1R) expression were not significantly altered by AT. HEK-293 cells were used to determine the potential humoral factors involved in down-regulation of NEP. Application of aldosterone, ANP, asymmetric dimethylarginine, and angiotensin peptides failed to cause down-regulation of renal NEP expression in vitro. AT reduces NEP expression and stimulates TGF-beta1 signalling in the kidneys. Thus, even brief episodes of

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

    PubMed

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

    2012-06-01

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

  9. Taxonomic recovery of the ant cricket Myrmecophilus albicinctus from M. americanus (Orthoptera, Myrmecophilidae)

    PubMed Central

    Komatsu, Takashi; Maruyama, Munetoshi

    2016-01-01

    Abstract Myrmecophilus americanus and Myrmecophilus albicinctus are typical myrmecophilous insects living inside ant nests. These species are ecologically important due to the obligate association with tramp ant species, including harmful invasive ant species. However, the taxonomy of these “white-banded ant crickets” is quite confused owing to a scarcity of useful external morphological characteristics. Recently, Myrmecophilus albicinctus was synonymized with Myrmecophilus americanus regardless of the apparent host use difference. To clarify taxonomical relationship between Myrmecophilus albicinctus and Myrmecophilus albicinctus, we reexamined morphological characteristics of both species mainly in the viewpoint of anatomy. Observation of genitalia parts, together with a few external body parts, revealed that Myrmecophilus albicinctus showed different tendency from them of Myrmecophilus americanus. Therefore, we recover Myrmecophilus albicinctus as a distinct species on the basis of the morphology. PMID:27408536

  10. Characterization and functional classification of American lobster (Homarus americanus) immune factor transcripts.

    PubMed

    Clark, K Fraser

    2014-11-01

    The American lobster (Homarus americanus) is the most important commercially exploited marine species in Canada. Very little is known about the H. americanus molecular humoral immune response or how to determine if a seemingly healthy lobster is infected with a pathogen. The goal of this work is to characterize several important H. americanus immune genes as well as highlight and classify hundreds of others into functional immune groups. The protein sequence of H. americanus acute phase serum amyloid protein A (SAA) was found to be similar to that of vertebrate SAA, and is likely a good clinical marker for immune activation in lobsters and some crustaceans. Additionally, only one gene, Trypsin 1b, was found to be differentially regulated during bacterial, microparasitic and viral challenges in lobster and is likely critical for the activation of the H. americanus immune response. Bioinformatic analysis was used to functionally annotate, 263 H. americanus immune genes and identify the few shared patterns of differential gene expression in lobsters in response to bacterial, parasitic and viral challenge. Many of the described immune genes are biomarker candidates which could be used as clinical indicators for lobster health and disease. Biomarkers can facilitate early detection of pathogens, or anthropomorphic stressors, so that mitigation strategies can be developed in order to prevent the devastating economic losses that have occurred in Southern New England, USA. This work is contributes to further our understanding of how the lobster immune system works and how it can be used to maintain the health and sustainability of the overall American lobster fishery.

  11. Neuropeptidergic Signaling in the American Lobster Homarus americanus: New Insights from High-Throughput Nucleotide Sequencing.

    PubMed

    Christie, Andrew E; Chi, Megan; Lameyer, Tess J; Pascual, Micah G; Shea, Devlin N; Stanhope, Meredith E; Schulz, David J; Dickinson, Patsy S

    2015-01-01

    Peptides are the largest and most diverse class of molecules used for neurochemical communication, playing key roles in the control of essentially all aspects of physiology and behavior. The American lobster, Homarus americanus, is a crustacean of commercial and biomedical importance; lobster growth and reproduction are under neuropeptidergic control, and portions of the lobster nervous system serve as models for understanding the general principles underlying rhythmic motor behavior (including peptidergic neuromodulation). While a number of neuropeptides have been identified from H. americanus, and the effects of some have been investigated at the cellular/systems levels, little is currently known about the molecular components of neuropeptidergic signaling in the lobster. Here, a H. americanus neural transcriptome was generated and mined for sequences encoding putative peptide precursors and receptors; 35 precursor- and 41 receptor-encoding transcripts were identified. We predicted 194 distinct neuropeptides from the deduced precursor proteins, including members of the adipokinetic hormone-corazonin-like peptide, allatostatin A, allatostatin C, bursicon, CCHamide, corazonin, crustacean cardioactive peptide, crustacean hyperglycemic hormone (CHH), CHH precursor-related peptide, diuretic hormone 31, diuretic hormone 44, eclosion hormone, FLRFamide, GSEFLamide, insulin-like peptide, intocin, leucokinin, myosuppressin, neuroparsin, neuropeptide F, orcokinin, pigment dispersing hormone, proctolin, pyrokinin, SIFamide, sulfakinin and tachykinin-related peptide families. While some of the predicted peptides are known H. americanus isoforms, most are novel identifications, more than doubling the extant lobster neuropeptidome. The deduced receptor proteins are the first descriptions of H. americanus neuropeptide receptors, and include ones for most of the peptide groups mentioned earlier, as well as those for ecdysis-triggering hormone, red pigment concentrating hormone

  12. Neuropeptidergic Signaling in the American Lobster Homarus americanus: New Insights from High-Throughput Nucleotide Sequencing

    PubMed Central

    Christie, Andrew E.; Chi, Megan; Lameyer, Tess J.; Pascual, Micah G.; Shea, Devlin N.; Stanhope, Meredith E.; Schulz, David J.; Dickinson, Patsy S.

    2015-01-01

    Peptides are the largest and most diverse class of molecules used for neurochemical communication, playing key roles in the control of essentially all aspects of physiology and behavior. The American lobster, Homarus americanus, is a crustacean of commercial and biomedical importance; lobster growth and reproduction are under neuropeptidergic control, and portions of the lobster nervous system serve as models for understanding the general principles underlying rhythmic motor behavior (including peptidergic neuromodulation). While a number of neuropeptides have been identified from H. americanus, and the effects of some have been investigated at the cellular/systems levels, little is currently known about the molecular components of neuropeptidergic signaling in the lobster. Here, a H. americanus neural transcriptome was generated and mined for sequences encoding putative peptide precursors and receptors; 35 precursor- and 41 receptor-encoding transcripts were identified. We predicted 194 distinct neuropeptides from the deduced precursor proteins, including members of the adipokinetic hormone-corazonin-like peptide, allatostatin A, allatostatin C, bursicon, CCHamide, corazonin, crustacean cardioactive peptide, crustacean hyperglycemic hormone (CHH), CHH precursor-related peptide, diuretic hormone 31, diuretic hormone 44, eclosion hormone, FLRFamide, GSEFLamide, insulin-like peptide, intocin, leucokinin, myosuppressin, neuroparsin, neuropeptide F, orcokinin, pigment dispersing hormone, proctolin, pyrokinin, SIFamide, sulfakinin and tachykinin-related peptide families. While some of the predicted peptides are known H. americanus isoforms, most are novel identifications, more than doubling the extant lobster neuropeptidome. The deduced receptor proteins are the first descriptions of H. americanus neuropeptide receptors, and include ones for most of the peptide groups mentioned earlier, as well as those for ecdysis-triggering hormone, red pigment concentrating hormone

  13. Morphological Dependence of Element Stoichiometry in the H. americanus Exoskeleton

    NASA Astrophysics Data System (ADS)

    Mergelsberg, S. T.; Ulrich, R. N.; Dove, P. M.

    2016-02-01

    The crustacean exoskeleton is a complex biocomposite of inorganic mineral and organic macromolecules that expresses highly divergent morphologies across different taxa. While the structures and compositions of the organic framework show complex links to environmental and developmental pressures, little is known about the mineral chemistry. Previous studies of the cuticle have assumed that magnesium, phosphorous, and other trace metals are largely contained in the inorganic mineral fraction. Due to analytical limitations of structural analyses and in situ spectroscopic methods, the stoichiometry of the organic and inorganic portions could not be resolved. For example, previous Raman and XRD studies conclude the higher concentrations of trace elements, such as P and Mg measured in reinforced structures, e.g. the claw and abdomen, are primarily determined by the mineral fraction. Using the American Lobster (Homarus americanus) as a model organism to establish relationships between body part function and cuticle composition, this study quantified the distributions of Mg and P in the mineral and organic fractions. The experiments were designed to dissolve the exoskeleton of 10 body parts using three types of solutions that were specific to extracting 1) the mineral phase, 2) protein, and 3) polysaccharide. Analysis of the solutions by ICP-OES shows the mineral phase contains magnesium and phosphorous at concentrations sufficient to support the formation of calcium-magnesium and phosphate minerals. The protein fraction of the body parts contains significantly more Mg and P than previously hypothesized, while the levels of P contained in the organic portion are fairly constant. The findings demonstrate the lobster cuticle contains a significant amount of non-mineralized P and Mg that is readily water-soluble in the protein component. However, for those body parts used for defense and food acquisition, such as the claw, the mineral component determines the overall

  14. Blindness in a wild American black bear cub (Ursus americanus).

    PubMed

    Dombrowski, Elizabeth; McGregor, Glenna F; Bauer, Bianca S; Parker, Dennilyn; Grahn, Bruce H

    2016-07-01

    An approximately six-month-old wild American black bear (Ursus americanus) was found wandering in Saskatchewan and was presented to the Veterinary Medical Centre of the Western College of Veterinary Medicine for apparent blindness. Clinical examination confirmed an inability to navigate a photopic maze, bilateral tapetal hyper-reflectivity, fundi devoid of retinal vessels, and small pale optic nerve papillae. Single-flash electroretinography revealed A and B-wave amplitudes of approximately 40 and 140 microvolts, respectively, in both eyes. Histologic abnormalities included bilateral optic papillary mineralization and bilateral segmental optic nerve degeneration, with occasional intralesional lymphocytes confirmed with immunohistochemistry for CD3+. There was also bilateral multifocal retinal dysplasia, gliosis, lymphocytic retinitis, a complete lack of retinal blood vessels, an intravitreal vascular membrane, and a mild lymphocytic-plasmacytic uveitis with small pre-iridal cellular membranes. The presence of a positive ERG in a blind bear with numerous retinal ganglion cells and degenerative changes in the optic nerve are most consistent with vision loss due to optic nerve injury, which given the young age of the bear likely occurred during ocular development. The presence of ocular inflammation suggests this injury resulted from an inflammatory/infectious process. The etiology could not be determined. Hepatic concentrations of vitamin A were within the normal reference range for domestic species. Pan-herpesvirus PCR and immunohistochemistry for canine distemper virus and Toxoplasma gondii were negative, although this does not rule out these or other infectious etiologies. This represents the first case report of neonatal or congenital ocular abnormalities in an ursid species.

  15. Immunizations with chimeric hepatitis B virus-like particles to induce potential anti-hepatitis C virus neutralizing antibodies.

    PubMed

    Vietheer, Patricia T K; Boo, Irene; Drummer, Heidi E; Netter, Hans-Jürgen

    2007-01-01

    Virus-like particles (VLPs) are highly immunogenic and proven to induce protective immunity. The small surface antigen (HBsAg-S) of hepatitis B virus (HBV) self-assembles into VLPs and its use as a vaccine results in protective antiviral immunity against HBV infections. Chimeric HBsAg-S proteins carrying foreign epitopes allow particle formation and have the ability to induce anti-foreign humoral and cellular immune responses. The insertion of the hypervariable region 1 (HVR1) sequence derived from the envelope protein 2 (E2) of hepatitis C virus (HCV) into the major antigenic site of HBsAg-S ('a'-determinant) resulted in the formation of highly immunogenic VLPs that retained the antigenicity of the inserted HVR1 sequence. BALB/c mice were immunized with chimeric VLPs, which resulted in antisera with anti-HCV activity. The antisera were able to immunoprecipitate native HCV envelope complexes (E1E2) containing homologous or heterologous HVR1 sequences. HCV E1E2 pseudotyped HIV-1 particles (HCVpp) were used to measure entry into HuH-7 target cells in the presence or absence of antisera that were raised against chimeric VLPs. Anti-HVR1 VLP sera interfered with entry of entry-competent HCVpps containing either homologous or heterologous HVR1 sequences. Also, immunizations with chimeric VLPs induced antisurface antigen (HBsAg) antibodies, indicating that HBV-specific antigenicity and immunogenicity of the 'a'-determinant region is retained. A multivalent vaccine against different pathogens based on the HBsAg delivery platform should be possible. We hypothesize that custom design of VLPs with an appropriate set of HCV-neutralizing epitopes will induce antibodies that would serve to decrease the viral load at the initial infecting inoculum.

  16. Ebola virus-like particles produced in insect cells exhibit dendritic cell stimulating activity and induce neutralizing antibodies

    SciTech Connect

    Ye Ling; Lin Jianguo; Sun Yuliang; Bennouna, Soumaya; Lo, Michael; Wu Qingyang; Bu Zhigao; Pulendran, Bali; Compans, Richard W. . E-mail: compans@microbio.emory.edu; Yang Chinglai . E-mail: chyang@emory.edu

    2006-08-01

    Recombinant baculoviruses (rBV) expressing Ebola virus VP40 (rBV-VP40) or GP (rBV-GP) proteins were generated. Infection of Sf9 insect cells by rBV-VP40 led to assembly and budding of filamentous particles from the cell surface as shown by electron microscopy. Ebola virus-like particles (VLPs) were produced by coinfection of Sf9 cells with rBV-VP40 and rBV-GP, and incorporation of Ebola GP into VLPs was demonstrated by SDS-PAGE and Western blot analysis. Recombinant baculovirus infection of insect cells yielded high levels of VLPs, which were shown to stimulate cytokine secretion from human dendritic cells similar to VLPs produced in mammalian cells. The immunogenicity of Ebola VLPs produced in insect cells was evaluated by immunization of mice. Analysis of antibody responses showed that most of the GP-specific antibodies were of the IgG2a subtype, while no significant level of IgG1 subtype antibodies specific for GP was induced, indicating the induction of a Th1-biased immune response. Furthermore, sera from Ebola VLP immunized mice were able to block infection by Ebola GP pseudotyped HIV virus in a single round infection assay, indicating that a neutralizing antibody against the Ebola GP protein was induced. These results show that production of Ebola VLPs in insect cells using recombinant baculoviruses represents a promising approach for vaccine development against Ebola virus infection.

  17. Transintestinal Cholesterol Transport Is Active in Mice and Humans and Controls Ezetimibe-Induced Fecal Neutral Sterol Excretion.

    PubMed

    Jakulj, Lily; van Dijk, Theo H; de Boer, Jan Freark; Kootte, Ruud S; Schonewille, Marleen; Paalvast, Yared; Boer, Theo; Bloks, Vincent W; Boverhof, Renze; Nieuwdorp, Max; Beuers, Ulrich H W; Stroes, Erik S G; Groen, Albert K

    2016-12-13

    Except for conversion to bile salts, there is no major cholesterol degradation pathway in mammals. Efficient excretion from the body is therefore a crucial element in cholesterol homeostasis. Yet, the existence and importance of cholesterol degradation pathways in humans is a matter of debate. We quantified cholesterol fluxes in 15 male volunteers using a cholesterol balance approach. Ten participants repeated the protocol after 4 weeks of treatment with ezetimibe, an inhibitor of intestinal and biliary cholesterol absorption. Under basal conditions, about 65% of daily fecal neutral sterol excretion was bile derived, with the remainder being contributed by direct transintestinal cholesterol excretion (TICE). Surprisingly, ezetimibe induced a 4-fold increase in cholesterol elimination via TICE. Mouse studies revealed that most of ezetimibe-induced TICE flux is mediated by the cholesterol transporter Abcg5/Abcg8. In conclusion, TICE is active in humans and may serve as a novel target to stimulate cholesterol elimination in patients at risk for cardiovascular disease. Copyright © 2016 Elsevier Inc. All rights reserved.

  18. Inhibition of TGF-β with neutralizing antibodies prevents radiation-induced acceleration of metastatic cancer progression

    PubMed Central

    Biswas, Swati; Guix, Marta; Rinehart, Cammie; Dugger, Teresa C.; Chytil, Anna; Moses, Harold L.; Freeman, Michael L.; Arteaga, Carlos L.

    2007-01-01

    We investigated whether TGF-β induced by anticancer therapies accelerates tumor progression. Using the MMTV/PyVmT transgenic model of metastatic breast cancer, we show that administration of ionizing radiation or doxorubicin caused increased circulating levels of TGF-β1 as well as increased circulating tumor cells and lung metastases. These effects were abrogated by administration of a neutralizing pan–TGF-β antibody. Circulating polyomavirus middle T antigen–expressing tumor cells did not grow ex vivo in the presence of the TGF-β antibody, suggesting autocrine TGF-β is a survival signal in these cells. Radiation failed to enhance lung metastases in mice bearing tumors that lack the type II TGF-β receptor, suggesting that the increase in metastases was due, at least in part, to a direct effect of TGF-β on the cancer cells. These data implicate TGF-β induced by anticancer therapy as a prometastatic signal in tumor cells and provide a rationale for the simultaneous use of these therapies in combination with TGF-β inhibitors. PMID:17415413

  19. Geographical venom variations of the Southeast Asian monocled cobra (Naja kaouthia): venom-induced neuromuscular depression and antivenom neutralization.

    PubMed

    Tan, Kae Yi; Tan, Choo Hock; Sim, Si Mui; Fung, Shin Yee; Tan, Nget Hong

    2016-01-01

    The Southeast Asian monocled cobras (Naja kaouthia) exhibit geographical variations in their venom proteomes, especially on the composition of neurotoxins. This study compared the neuromuscular depressant activity of the venoms of N. kaouthia from Malaysia (NK-M), Thailand (NK-T) and Vietnam (NK-V), and the neutralization of neurotoxicity by a monospecific antivenom. On chick biventer cervicis nerve-muscle preparation, all venoms abolished the indirect twitches, with NK-T venom being the most potent (shortest t90, time to 90% twitch inhibition), followed by NK-V and NK-M. Acetylcholine and carbachol failed to reverse the blockade, indicating irreversible/pseudo-irreversible post-synaptic neuromuscular blockade. KCl restored the twitches variably (NK-M preparation being the least responsive), consistent with different degree of muscle damage. The findings support that NK-T venom has the most abundant curarimimetic alpha-neurotoxins, while NK-M venom contains more tissue-damaging cytotoxins. Pre-incubation of tissue with N. kaouthia monovalent antivenom (NKMAV) prevented venom-induced twitch depression, with the NK-T preparation needing the largest antivenom dose. NKMAV added after the onset of neuromuscular depression could only halt the inhibitory progression but failed to restore full contraction. The findings highlight the urgency of early antivenom administration to sequester as much circulating neurotoxins as possible, thereby hastening toxin elimination from the circulation. In envenomed mice, NKMAV administered upon the first neurological sign neutralized the neurotoxic effect, with the slowest full recovery noticed in the NK-T group. This is consistent with the high abundance of neurotoxins in the NK-T venom, implying that a larger amount or repeated dosing of NKMAV may be required in NK-T envenomation. Copyright © 2016 Elsevier Inc. All rights reserved.

  20. Inhibition of neutral sphingomyelinase decreases elevated levels of inducible nitric oxide synthase and apoptotic cell death in ocular hypertensive rats

    SciTech Connect

    Aslan, Mutay; Basaranlar, Goksun; Unal, Mustafa; Ciftcioglu, Akif; Derin, Narin; Mutus, Bulent

    2014-11-01

    Endoplasmic reticulum (ER) stress and excessive nitric oxide production via induction of inducible nitric oxide synthase (NOS2) have been implicated in the pathogenesis of neuronal retinal cell death in ocular hypertension. Neutral sphingomyelinase (N-SMase)/ceramide pathway can regulate NOS2 expression, hence this study determined the role of selective neutral sphingomyelinase (N-SMase) inhibition on retinal NOS2 levels, ER stress, apoptosis and visual evoked potentials (VEPs) in a rat model of elevated intraocular pressure (EIOP). NOS2 expression and retinal protein nitration were significantly greater in EIOP and significantly decreased with N-SMase inhibition. A significant increase was observed in retinal ER stress markers pPERK, CHOP and GRP78 in EIOP, which were not significantly altered by N-SMase inhibition. Retinal TUNEL staining showed increased apoptosis in all EIOP groups; however N-SMase inhibition significantly decreased the percent of apoptotic cells in EIOP. Caspase-3, -8 and -9 activities were significantly increased in EIOP and returned to baseline levels following N-SMase inhibition. Latencies of all VEP components were significantly prolonged in EIOP and shortened following N-SMase inhibition. Data confirm the role of nitrative injury in EIOP and highlight the protective effect of N-SMase inhibition in EIOP via down-regulation of NOS2 levels and nitrative stress. - Highlights: • Inhibition of N-SMase decreases NOS2 levels in ocular hypertension. • Inhibition of N-SMase decreases protein nitration in ocular hypertension. • Inhibition of N-SMase decreases caspase activation in ocular hypertension. • Inhibition of N-SMase decreases apoptosis in ocular hypertension.

  1. Molecular dynamics simulation of disorder-induced heating in ultracold neutral plasma.

    PubMed

    Guo, L; Lu, R H; Han, S S

    2010-04-01

    Disorder-induced heating (DIH) is one of the main reasons reducing the coupling strength in ultracold plasma. We propose applying an optical lattice as periodic confinement before the ultracold atomic cloud is ionized to eliminate its effect. We demonstrate a numerical simulation for the dynamics of the ultracold plasmas using classical molecular dynamics method with open boundary. DIH is reproduced in the simulation with the random Gaussian initial distribution and is absent in the results with the ordered lattice initial distribution. We further find that the collisional heating from electrons is important for ultracold plasmas with chosen spatial correlations in the optical lattice. Carefully preparing the initial condition (e.g., the ion density, initial electron temperature, and so on), collisional heating for the ions would be significantly reduced and eventually negligible. This allows a much stronger coupling in ultracold plasma to be realized.

  2. Broadening the Heterologous Cross-Neutralizing Antibody Inducing Ability of Porcine Reproductive and Respiratory Syndrome Virus by Breeding the GP4 or M genes

    PubMed Central

    Zhou, Lei; Ni, Yan-Yan; Piñeyro, Pablo; Cossaboom, Caitlin M.; Subramaniam, Sakthivel; Sanford, Brenton J.; Dryman, Barbara A.; Huang, Yao-Wei; Meng, Xiang-Jin

    2013-01-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most economically important swine pathogens, which causes reproductive failure in sows and respiratory disease in piglets. A major hurdle to control PRRSV is the ineffectiveness of the current vaccines to confer protection against heterologous strains. Since both GP4 and M genes of PRRSV induce neutralizing antibodies, in this study we molecularly bred PRRSV through DNA shuffling of the GP4 and M genes, separately, from six genetically different strains of PRRSV in an attempt to identify chimeras with improved heterologous cross-neutralizing capability. The shuffled GP4 and M genes libraries were each cloned into the backbone of PRRSV strain VR2385 infectious clone pIR-VR2385-CA. Three GP4-shuffled chimeras and five M-shuffled chimeras, each representing sequences from all six parental strains, were selected and further characterized in vitro and in pigs. These eight chimeric viruses showed similar levels of replication with their backbone strain VR2385 both in vitro and in vivo, indicating that the DNA shuffling of GP4 and M genes did not significantly impair the replication ability of these chimeras. Cross-neutralization test revealed that the GP4-shuffled chimera GP4TS14 induced significantly higher cross-neutralizing antibodies against heterologous strains FL-12 and NADC20, and similarly that the M-shuffled chimera MTS57 also induced significantly higher levels of cross-neutralizing antibodies against heterologous strains MN184B and NADC20, when compared with their backbone parental strain VR2385 in infected pigs. The results suggest that DNA shuffling of the GP4 or M genes from different parental viruses can broaden the cross-neutralizing antibody-inducing ability of the chimeric viruses against heterologous PRRSV strains. The study has important implications for future development of a broadly protective vaccine against PRRSV. PMID:23826108

  3. Broadening the heterologous cross-neutralizing antibody inducing ability of porcine reproductive and respiratory syndrome virus by breeding the GP4 or M genes.

    PubMed

    Zhou, Lei; Ni, Yan-Yan; Piñeyro, Pablo; Cossaboom, Caitlin M; Subramaniam, Sakthivel; Sanford, Brenton J; Dryman, Barbara A; Huang, Yao-Wei; Meng, Xiang-Jin

    2013-01-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most economically important swine pathogens, which causes reproductive failure in sows and respiratory disease in piglets. A major hurdle to control PRRSV is the ineffectiveness of the current vaccines to confer protection against heterologous strains. Since both GP4 and M genes of PRRSV induce neutralizing antibodies, in this study we molecularly bred PRRSV through DNA shuffling of the GP4 and M genes, separately, from six genetically different strains of PRRSV in an attempt to identify chimeras with improved heterologous cross-neutralizing capability. The shuffled GP4 and M genes libraries were each cloned into the backbone of PRRSV strain VR2385 infectious clone pIR-VR2385-CA. Three GP4-shuffled chimeras and five M-shuffled chimeras, each representing sequences from all six parental strains, were selected and further characterized in vitro and in pigs. These eight chimeric viruses showed similar levels of replication with their backbone strain VR2385 both in vitro and in vivo, indicating that the DNA shuffling of GP4 and M genes did not significantly impair the replication ability of these chimeras. Cross-neutralization test revealed that the GP4-shuffled chimera GP4TS14 induced significantly higher cross-neutralizing antibodies against heterologous strains FL-12 and NADC20, and similarly that the M-shuffled chimera MTS57 also induced significantly higher levels of cross-neutralizing antibodies against heterologous strains MN184B and NADC20, when compared with their backbone parental strain VR2385 in infected pigs. The results suggest that DNA shuffling of the GP4 or M genes from different parental viruses can broaden the cross-neutralizing antibody-inducing ability of the chimeric viruses against heterologous PRRSV strains. The study has important implications for future development of a broadly protective vaccine against PRRSV.

  4. A novel “correlated ion and neutral time of flight” method: Event-by-event detection of neutral and charged fragments in collision induced dissociation of mass selected ions

    SciTech Connect

    Teyssier, C.; Fillol, R.; Abdoul-Carime, H.; Farizon, B.; Farizon, M.

    2014-01-15

    A new tandem mass spectrometry (MS/MS) method based on time of flight measurements performed on an event-by-event detection technique is presented. This “correlated ion and neutral time of flight” method allows to explore Collision Induced Dissociation (CID) fragmentation processes by directly identifying not only all ions and neutral fragments produced but also their arrival time correlations within each single fragmentation event from a dissociating molecular ion. This constitutes a new step in the characterization of molecular ions. The method will be illustrated here for a prototypical case involving CID of protonated water clusters H{sup +}(H{sub 2}O){sub n=1–5} upon collisions with argon atoms.

  5. Laser Induced Fluorescence (LIF) Measurement System for Neutral Argon (Ar I) Dynamics in the HelCat Plasma Device at UNM

    NASA Astrophysics Data System (ADS)

    Kelly, Ralph; Gilmore, Mark; Lynn, Alan; Desjardins, Tiffany; Boynton, Nick; Nguyen, Luan; Scruggs, Andrew; Romero, Jorge

    2015-11-01

    When neutral and ion densities are spatially nonuniform, neutral-ion collisions can exert a torque on a magnetized plasma column via the FxB force, where F is the force exerted on ions by neutrals. This FxB force may have a significant effect on the dynamics of plasma instabilities and flows. In order to investigate the role of neutral dynamics in helicon discharges in the HelCat (Helicon-Cathode) plasma device at U. New Mexico, an Ar I Laser Induced Fluorescence (LIF) system is being developed. Previous passive spectroscopic measurements of Ar I and Ar II lines indicate that the neutral density profile is hollow (higher nn at larger radius). Additionally, we have not been able to reconcile azimuthal flows measured by Mach probes with those expected from ExB and diamagnetic torques. It is hypothesized that neutrals play an important role in the plasma flow. The LIF system is based on a >250 mW, tunable solid state laser. The laser will pump the metastable (2P3/2 0) 4s2 level to the (2P1/2 0) 4p2 level using 696.543 nm light, and observe fluorescence radiation from decay to the (2P1/2 0) 4s2 level at 772.42 nm. The system design and initial results will be presented. Work supported by NSF.

  6. Activity of Ca2+-dependent neutral proteases in tissues of ground squirrel during hibernation and during self-warming after induced awakening.

    PubMed

    Nurmagomedova, P M; Abasova, M M; Emirbekov, E Z

    2011-09-01

    Cyclic changes in activity of Ca2+-dependent neutral protease occur during preparation for hibernation, with an increase in September and November and decrease in October and December. During hibernation proteolytic enzyme activity decreased, while during self-warming after induced awakening, the role of Ca2+-dependent processes in the tissues of ground squirrels increased according to the body temperature.

  7. Molecular characterization of a novel soybean gene encoding a neutral PR-5 protein induced by high-salt stress.

    PubMed

    Tachi, Hiroyuki; Fukuda-Yamada, Kumiko; Kojima, Toshio; Shiraiwa, Masakazu; Takahara, Hidenari

    2009-01-01

    In this study, we characterized a novel soybean gene encoding a neutral PR-5 protein and compared it to two acidic isoforms of soybean PR-5 protein. This gene, designated as Glycine max osmotin-like protein, b isoform (GmOLPb, accession no. AB370233), encoded a putative protein having the greatest similarity to chickpea PR-5b (89% identity). Unlike the two acidic PR-5, GmOLPa and P21, the protein had a C-terminal elongation responsible for possible vacuolar targeting and after maturation showed a calculated molecular mass of 21.9kDa with pI 6.0. The 3D models, predicted by the homology modeling, contained four alpha-helixes and 16 beta-strands and formed three characteristic domains. The two acidic PR-5 proteins also showed a 3D structure very similar to GmOLPb, although the electrostatic potential on molecular surface of each PR-5 was significantly different. In the study of the gene expression under conditions of high-salt stress, GmOLPb was highly induced in the leaves of the soybean, particularly in the lower part of a leaf. The expression started at 2h after initiation of the stress and was highly induced between 18-72h. Gene expression of P21e (protein homologous to P21) was transiently induced by high-salt stress, but took place earlier than the gene expressions of GmOLPa and GmOLPb. Such differential expression was observed also under investigation with methyl jasmonate and salicylic acid. These results suggested that each soybean PR-5 might play a distinctive role in the defensive system protecting the soybean plant against high-salt stress, particularly in the leaves of the soybean.

  8. Japanese encephalitis protein vaccine candidates expressing neutralizing epitope and M.T hsp70 induce virus-specific memory B cells and long-lasting antibodies in swine.

    PubMed

    Fei-fei, Ge; Jian, Wang; Feng, Xu; Li-ping, Sheng; Quan-yun, Sun; Jin-ping, Zhou; Pu-yan, Chen; Pei-hong, Liu

    2008-10-16

    Swine are an important amplifier of Japanese encephalitis (JE) virus in the paradomestic environment. In this study, two JE protein vaccine candidates were evaluated for immunogenicity in swine. Both vaccine plasmids are based on a prokaryotic vector pET-32a(+). One plasmid, designated pET-32a(+)-epitope, encode a cassette consisting of a neutralizing epitope on envelope (E) protein of JE virus, whereas the other plasmid, designated pET-32a(+)-epitope-hsp70, express the fusion protein of the epitope and M.T hsp70. Some differences were detected in the immunogenicity of these two proteins in swine. Swine immunized twice with 2000pmol of the neutralizing epitope or the fusion protein developed neutralizing antibody titers of respectively, 154 and 300, and anti-neutralizing epitope antibody titers of 10(4.25) and 10(6.0) by 3 weeks after the second immunization. In addition, swine immunized with the neutralizing epitope emulsified with adjuvant S206 or with imported mineral oil and Tween-80 induced neutralizing antibody titers of 196 and 244, and anti-neutralizing epitope antibody titers of 10(5.25) or 10(5.6) at the same time point. However, swine administered two doses of a commercial JE vaccine (attenuated virus preparation; JEV SA14-14-2 strain) developed less favorable antibody responses with neutralizing antibody titer 40 and anti-neutralizing epitope antibody titers 10(3.7). The anamnestic response was followed by monitoring titers 1 week after boosting with a viral antigen; swine immunized twice with the fusion protein showed a 177-fold increase in anti-neutralizing epitope titer, indicating a strong recall of the antibody response. The animals maintained detectable levels of anti-neutralizing epitope antibody for at least 105 days after two immunizations, indicating that these four protein antigens are able to stimulate virus-specific memory B cells and long-lasting antibodies at higher levels than is achieved using a current commercial attenuated JEV vaccine

  9. Simultaneous determination of neutral and acidic pharmaceuticals in wastewater by high-performance liquid chromatography-post-column photochemically induced fluorimetry.

    PubMed

    González-Barreiro, C; Lores, M; Casais, M C; Cela, R

    2003-04-18

    An analytical method for the simultaneous determination of acidic and neutral pharmaceutical active compound (PhACs) residues in wastewater has been developed based on the combination of high-performance liquid chromatography (HPLC) and photochemically induced fluorimetry. The photoderivatization conditions for each particular PhAC have been assessed. Off-line optimization of the HPLC separation for both neutral and acidic compounds has been utilised and evaluated. Detection limits in the low ng/ml range have been achieved without sample pretreatment. By applying the developed analytical method combined with solid-phase extraction to real wastewater samples an enrichment factor of approximately two orders of magnitude can be obtained.

  10. Nonneutralizing Antibodies Induced by the HIV-1 gp41 NHR Domain Gain Neutralizing Activity in the Presence of the HIV Fusion Inhibitor Enfuvirtide: a Potential Therapeutic Vaccine Strategy.

    PubMed

    Wang, Qian; Bi, Wenwen; Zhu, Xiaojie; Li, Haoyang; Qi, Qianqian; Yu, Fei; Lu, Lu; Jiang, Shibo

    2015-07-01

    A key barrier against developing preventive and therapeutic human immunodeficiency virus (HIV) vaccines is the inability of viral envelope glycoproteins to elicit broad and potent neutralizing antibodies. However, in the presence of fusion inhibitor enfuvirtide, we show that the nonneutralizing antibodies induced by the HIV type 1 (HIV-1) gp41 N-terminal heptad repeat (NHR) domain (N63) exhibit potent and broad neutralizing activity against laboratory-adapted HIV-1 strains, including the drug-resistant variants, and primary HIV-1 isolates with different subtypes, suggesting the potential of developing gp41-targeted HIV therapeutic vaccines.

  11. Seroprevalence of Toxoplasma gondii in American Black Bears ( Ursus americanus ) of the Central Appalachians, USA.

    PubMed

    Cox, John J; Murphy, Sean M; Augustine, Ben C; Guthrie, Joseph M; Hast, John T; Maehr, Sutton C; McDermott, Joseph

    2017-07-01

    We assessed Toxoplasma gondii seroprevalence in 53 free-ranging American black bears ( Ursus americanus ) in the Central Appalachian Mountains, US. Seroprevalence was 62% with no difference between males and females or between juvenile and adult bears. Wildlife agencies should consider warnings in hunter education programs to reduce the chances for human infection from this source.

  12. Prevalence of Anaplasma phagocytophilum in North Carolina Eastern Black Bears ( Ursus americanus ).

    PubMed

    Westmoreland, Lori S H; Stoskopf, Michael K; Maggi, Ricardo G

    2016-10-01

    We detected Anaplasma phagocytophilum by DNA amplification in whole blood from free-ranging, hunter-killed American black bears ( Ursus americanus ) from the east coast of North Carolina, US. Molecular prevalence for Anaplasma phagocytophilum was 3% from 68 black bears. No DNA of other Anaplasma or Ehrlichia spp. was identified.

  13. Draft Genome Sequence of "Candidatus Liberibacter americanus" Bacterium Associated with Citrus Huanglongbing in Brazil.

    PubMed

    Lin, Hong; Coletta-Filho, Helvecio D; Han, Cliff S; Lou, Binghai; Civerolo, Edwin L; Machado, Marcos A; Gupta, Goutam

    2013-05-23

    We report here the draft genome sequence of "Candidatus Liberibacter americanus" strain PW_SP. The 1,176,071-bp genome, with 31.6% G+C content, comprises 948 open reading frames, 38 tRNAs, and three complete rRNAs.

  14. Potential Vertical Transmission of Winter Ticks (Dermacentor albipictus) from Moose (Alces americanus) Dams to Neonates.

    PubMed

    Severud, William J; DelGiudice, Glenn D

    2016-01-01

    North American moose (Alces americanus) frequently become infested with winter ticks (Dermacentor albipictus). During capture of neonatal moose in northeastern Minnesota, US, in May-June 2013 and 2014, we recovered adult ticks from neonates, presumably vertically transferred from dams, heretofore, not documented. Infestations on neonates may have population-level implications.

  15. Stable Carbon and Oxygen Isotope Ratios of Otoliths Differentiate Winter Flounder (Pseudopleuonectes americanus) Habitats

    EPA Science Inventory

    Stable carbon (13C) and oxygen (18O) isotope ratios were measured in otoliths of juvenile winter flounder (Pseudopleuronectes americanus) collected from 18 nursery areas along the coast of Rhode Island, USA. Samples were obtained during June and July of 2002 from locations tha...

  16. Stable Carbon and Oxygen Isotope Ratios of Otoliths Differentiate Winter Flounder (Pseudopleuonectes americanus) Habitats

    EPA Science Inventory

    Stable carbon (13C) and oxygen (18O) isotope ratios were measured in otoliths of juvenile winter flounder (Pseudopleuronectes americanus) collected from 18 nursery areas along the coast of Rhode Island, USA. Samples were obtained during June and July of 2002 from locations tha...

  17. Movements, activity patterns, and habitat use of snowshoe hares (Lepus americanus) in interior Alaska

    Treesearch

    D. Feierabend; K. Kielland

    2014-01-01

    Snowshoe hares (Lepus americanus) are generally sedentary, but are likely to move among habitats frequently to gain access to spatially segregated food and cover. We investigated movement patterns of hares from 2 characteristic boreal habitats using very-high-frequency radio collars (n ¼ 300) monitored weekly and global positioning system (GPS) collars (n ¼ 18)...

  18. SARCOCYSTIS URSUSI, N. SP (APICOMPLEXA; SARCOCYSTIDAE) FROM THE BLACK BEAR (URSUS AMERICANUS)

    USDA-ARS?s Scientific Manuscript database

    Infection with Sarcocystis species is common in hervibores but is rare in bears. Histological sections of 374 black bears (Ursus americanus) from Pennsylvania were examined for sarcocysts. A total of 3 sarcocysts were found in 3 bears, 1 sarcocyst per section. Sarcocysts from 2 bears were considered...

  19. GM-CSF in murine psoriasiform dermatitis: Redundant and pathogenic roles uncovered by antibody-induced neutralization and genetic deficiency

    PubMed Central

    Scholz, Tatjana; Weigert, Andreas; Brüne, Bernhard; Sadik, Christian D.; Böhm, Beate

    2017-01-01

    Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic, Th17-derived cytokine thought to critically contribute to the pathogenesis of diverse autoimmune diseases, including rheumatoid arthritis and psoriasis. Treatment with monoclonal antibodies that block GM-CSF activity is associated with favorable therapeutic effects in patients with rheumatoid arthritis. We evaluated the role of GM-CSF as a potential target for therapeutic interference in psoriasis using a combined pharmacologic and genetic approach and the mouse model of imiquimod-induced psoriasiform dermatitis (IMQPD). Neutralization of murine GM-CSF by an anti-GM-CSF antibody ameliorated IMQPD. In contrast, genetic deficiency in GM-CSF did not alter the course of IMQPD, suggesting the existence of mechanisms compensating for chronic, but not acute, absence of GM-CSF. Further investigation uncovered an alternative pathogenic pathway for IMQPD in the absence of GM-CSF characterized by an expanded plasmacytoid dendritic cell population and release of IFNα and IL-22. This pathway was not activated in wild-type mice during short-term anti-GM-CSF treatment. Our investigations support the potential value of GM-CSF as a therapeutic target in psoriatic disease. The discovery of an alternative pathogenic pathway for psoriasiform dermatitis in the permanent absence of GM-CSF, however, suggests the need for monitoring during therapeutic use of long-term GM-CSF blockade. PMID:28777803

  20. Nocardia-induced granulocyte macrophage colony-stimulating factor is neutralized by autoantibodies in disseminated/extrapulmonary nocardiosis.

    PubMed

    Rosen, Lindsey B; Rocha Pereira, Nuno; Figueiredo, Cristóvão; Fiske, Lauren C; Ressner, Roseanne A; Hong, Julie C; Gregg, Kevin S; Henry, Tracey L; Pak, Kirk J; Baumgarten, Katherine L; Seoane, Leonardo; Garcia-Diaz, Julia; Olivier, Kenneth N; Zelazny, Adrian M; Holland, Steven M; Browne, Sarah K

    2015-04-01

    Nocardia species cause infections in both immunocompromised and otherwise immunocompetent patients, although the mechanisms defining susceptibility in the latter group are elusive. Anticytokine autoantibodies are an emerging cause of pathogen-specific susceptibility in previously healthy human immunodeficiency virus-uninfected adults, including anti-granulocyte macrophage colony-stimulating factor (GM-CSF) autoantibodies with cryptococcal meningitis. Plasma from patients with disseminated/extrapulmonary nocardiosis and healthy controls was screened for anticytokine autoantibodies using a particle-based approach. Autoantibody function was assessed by intranuclear staining for GM-CSF-induced STAT5 phosphorylation in normal cells incubated with either patient or normal plasma. GM-CSF-mediated cellular activation by Nocardia was assessed by staining for intracellular cytokine production and intranuclear STAT5 phosphorylation. We identified neutralizing anti-GM-CSF autoantibodies in 5 of 7 patients studied with central nervous system nocardiosis and in no healthy controls (n = 14). GM-CSF production was induced by Nocardia in vitro, suggesting a causative role for anti-GM-CSF autoantibodies in Nocardia susceptibility and dissemination. In previously healthy adults with otherwise unexplained disseminated/extrapulmonary Nocardia infections, anti-GM-CSF autoantibodies should be considered. Their presence may suggest that these patients may be at risk for later development of pulmonary alveolar proteinosis or other opportunistic infections, and that patients may benefit from therapeutic GM-CSF administration. Published by Oxford University Press on behalf of the Infectious Diseases Society of America 2014. This work is written by (a) US Government employee(s) and is in the public domain in the US.

  1. Human Immunodeficiency Virus Type 1 gp120 Induces Apoptosis in Human Primary Neurons through Redox-Regulated Activation of Neutral Sphingomyelinase

    PubMed Central

    Jana, Arundhati; Pahan, Kalipada

    2007-01-01

    Human immunodeficiency virus type 1 (HIV-1) infection is known to cause disorders of the CNS, including HIV-associated dementia (HAD). HIV-1 coat protein gp120 (glycoprotein 120) induces neuronal apoptosis and has been implicated in the pathogenesis of HAD. However, the mechanism by which gp120 causes neuronal apoptosis is poorly understood. The present study underlines the importance of gp120 in inducing the production of ceramide, an important inducer of apoptosis, in human primary neurons. gp120 induced the activation of sphingomyelinases (primarily the neutral one) and the production of ceramide in primary neurons. Antisense knockdown of neutral (NSMase) but not acidic (ASMase) sphingomyelinase markedly inhibited gp120-mediated apoptosis and cell death of primary neurons, suggesting that the activation of NSMase but not ASMase plays an important role in gp120-mediated neuronal apoptosis. Similarly, the HIV-1 regulatory protein Tat also induced neuronal cell death via NSMase. Furthermore, gp120-induced production of ceramide was redox sensitive, because reactive oxygen species were involved in the activation of NSMase but not ASMase. gp120 coupled CXCR4 (CXC chemokine receptor 4) to induce NADPH oxidase-mediated production of superoxide radicals in neurons, which was involved in the activation of NSMase but not ASMase. These studies suggest that gp120 may induce neuronal apoptosis in the CNS of HAD patients through the CXCR4–NADPH oxidase–superoxide–NSMase–ceramide pathway. PMID:15509740

  2. Necator americanus Infection: A Possible Cause of Altered Dendritic Cell Differentiation and Eosinophil Profile in Chronically Infected Individuals

    PubMed Central

    Fujiwara, Ricardo T.; Cançado, Guilherme G. L.; Freitas, Paula A.; Santiago, Helton C.; Massara, Cristiano Lara; dos Santos Carvalho, Omar; Corrêa-Oliveira, Rodrigo; Geiger, Stefan M.; Bethony, Jeffrey

    2009-01-01

    Background Hookworms survive for several years (5 to 7 years) in the host lumen, inducing a robust but largely ineffective immune response. Among the most striking aspects of the immune response to hookworm (as with many other helminths) is the ablation of parasite-specific T cell proliferative response (hyporesponsiveness). While the role of the adaptive immune response in human helminth infection has been well investigated, the role of the innate immune responses (e.g., dendritic cells and eosinophils) has received less attention and remains to be clearly elucidated. Methodology/Principal Findings We report on the differentiation/maturation of host dendritic cells in vitro and the eosinophil activation/function associated with human hookworm infection. Mature DCs (mDCs) from Necator americanus (Necator)–infected individuals showed an impaired differentiation process compared to the mDCs of non-infected individuals, as evidenced by the differential expression of CD11c and CD14. These same hookworm-infected individuals also presented significantly down-regulated expression of CD86, CD1a, HLA-ABC, and HLA-DR. The lower expression of co-stimulatory and antigen presentation molecules by hookworm-infected–derived mDCs was further evidenced by their reduced ability to induce cell proliferation. We also showed that this alternative DC differentiation is partially induced by excreted-secreted hookworm products. Conversely, eosinophils from the same individuals showed a highly activated status, with an upregulation of major cell surface markers. Antigen-pulsed eosinophils from N. americanus–infected individuals induced significant cell proliferation of autologous PBMCs, when compared to non-infected individuals. Conclusion Chronic N. americanus infection alters the host's innate immune response, resulting in a possible modulation of the maturation process of DCs, a functional change that may diminish their ability for antigen presentation and thus contribute to the

  3. The Inhibitory Effects of Anti-Oxidants on Ultraviolet-Induced Up-Regulation of the Wrinkling-Inducing Enzyme Neutral Endopeptidase in Human Fibroblasts

    PubMed Central

    Nakajima, Hiroaki; Terazawa, Shuko; Niwano, Takao; Yamamoto, Yorihiro; Imokawa, Genji

    2016-01-01

    We recently reported that the over-expression of skin fibroblast-derived neutral endopeptidase (NEP) plays a pivotal role in impairing the three-dimensional architecture of dermal elastic fibers during the biological mechanism of ultraviolet (UV)-induced skin wrinkling. In that process, a UVB-associated epithelial-mesenchymal cytokine interaction as well as a direct UVA-induced cellular stimulation are associated with the up-regulation of NEP in human fibroblasts. In this study, we characterized the mode of action of ubiquinol10 which may abrogate the up-regulation of NEP by dermal fibroblasts, resulting in a reported in vivo anti-wrinkling action, and compared that with 3 other anti-oxidants, astaxanthin (AX), riboflavin (RF) and flavin mononucleotide (FMN). Post-irradiation treatment with all 4 of those anti-oxidants elicited an interrupting effect on the UVB-associated epithelial-mesenchymal cytokine interaction leading to the up-regulation of NEP in human fibroblasts but with different modes of action. While AX mainly served as an inhibitor of the secretion of wrinkle-inducing cytokines, such as interleukin-1α (IL-1α) and granulocyte macrophage colony stimulatory factor (GM-CSF) in UVB-exposed epidermal keratinocytes, ubiquinol10, RF and FMN predominantly interrupted the IL-1α and GM-CSF-stimulated expression of NEP in dermal fibroblasts. On the other hand, as for the UVA-associated mechanism, similar to the abrogating effects reported for AX and FMN, ubiquinol10 but not RF had the potential to abrogate the increased expression of NEP and matrix-metalloproteinase-1 in UVA-exposed human fibroblasts. Our findings strongly support the in vivo anti-wrinkling effects of ubiquinol10 and AX on human and animal skin and provide convincing proof of the UV-induced wrinkling mechanism that essentially focuses on the over-expression of NEP by dermal fibroblasts as an intrinsic causative factor. PMID:27648570

  4. Hepatitis C Virus Hypervariable Region 1 Variants Presented on Hepatitis B Virus Capsid-Like Particles Induce Cross-Neutralizing Antibodies

    PubMed Central

    Bankwitz, Dorothea; Osburn, William; Viazov, Sergei; Brovko, Olena; Zekri, Abdel-Rahman; Khudyakov, Yury; Nassal, Michael; Pumpens, Paul; Pietschmann, Thomas; Timm, Jörg; Roggendorf, Michael; Walker, Andreas

    2014-01-01

    Hepatitis C virus (HCV) infection is still a serious global health burden. Despite improved therapeutic options, a preventative vaccine would be desirable especially in undeveloped countries. Traditionally, highly conserved epitopes are targets for antibody-based prophylactic vaccines. In HCV-infected patients, however, neutralizing antibodies are primarily directed against hypervariable region I (HVRI) in the envelope protein E2. HVRI is the most variable region of HCV, and this heterogeneity contributes to viral persistence and has thus far prevented the development of an effective HVRI-based vaccine. The primary goal of an antibody-based HCV vaccine should therefore be the induction of cross-reactive HVRI antibodies. In this study we approached this problem by presenting selected cross-reactive HVRI variants in a highly symmetric repeated array on capsid-like particles (CLPs). SplitCore CLPs, a novel particulate antigen presentation system derived from the HBV core protein, were used to deliberately manipulate the orientation of HVRI and therefore enable the presentation of conserved parts of HVRI. These HVRI-CLPs induced high titers of cross-reactive antibodies, including neutralizing antibodies. The combination of only four HVRI CLPs was sufficient to induce antibodies cross-reactive with 81 of 326 (24.8%) naturally occurring HVRI peptides. Most importantly, HVRI CLPs with AS03 as an adjuvant induced antibodies with a 10-fold increase in neutralizing capability. These antibodies were able to neutralize infectious HCVcc isolates and 4 of 19 (21%) patient-derived HCVpp isolates. Taken together, these results demonstrate that the induction of at least partially cross-neutralizing antibodies is possible. This approach might be useful for the development of a prophylactic HCV vaccine and should also be adaptable to other highly variable viruses. PMID:25014219

  5. Diversity in the structures and ligand-binding sites of nematode fatty acid and retinol-binding proteins revealed by Na-FAR-1 from Necator americanus

    PubMed Central

    Rey-Burusco, M. Florencia; Ibáñez-Shimabukuro, Marina; Gabrielsen, Mads; Franchini, Gisela R.; Roe, Andrew J.; Griffiths, Kate; Zhan, Bin; Cooper, Alan; Kennedy, Malcolm W.; Córsico, Betina; Smith, Brian O.

    2015-01-01

    Fatty acid and retinol-binding proteins (FARs) comprise a family of unusual α-helix rich lipid-binding proteins found exclusively in nematodes. They are secreted into host tissues by parasites of plants, animals and humans. The structure of a FAR protein from the free-living nematode Caenorhabditis elegans is available, but this protein [C. elegans FAR-7 (Ce-FAR-7)] is from a subfamily of FARs that does not appear to be important at the host/parasite interface. We have therefore examined [Necator americanus FAR-1 (Na-FAR-1)] from the blood-feeding intestinal parasite of humans, N. americanus. The 3D structure of Na-FAR-1 in its ligand-free and ligand-bound forms, determined by NMR (nuclear magnetic resonance) spectroscopy and X-ray crystallography respectively, reveals an α-helical fold similar to Ce-FAR-7, but Na-FAR-1 possesses a larger and more complex internal ligand-binding cavity and an additional C-terminal α-helix. Titration of apo-Na-FAR-1 with oleic acid, analysed by NMR chemical shift perturbation, reveals that at least four distinct protein–ligand complexes can be formed. Na-FAR-1 and possibly other FARs may have a wider repertoire for hydrophobic ligand binding, as confirmed in the present study by our finding that a range of neutral and polar lipids co-purify with the bacterially expressed recombinant protein. Finally, we show by immunohistochemistry that Na-FAR-1 is present in adult worms with a tissue distribution indicative of possible roles in nutrient acquisition by the parasite and in reproduction in the male. PMID:26318523

  6. Formalin-inactivated EV71 vaccine candidate induced cross-neutralizing antibody against subgenotypes B1, B4, B5 and C4A in adult volunteers.

    PubMed

    Chou, Ai-Hsiang; Liu, Chia-Chyi; Chang, Jui-Yuan; Jiang, Renee; Hsieh, Yi-Chin; Tsao, Amanda; Wu, Chien-Long; Huang, Ju-Lan; Fung, Chang-Phone; Hsieh, Szu-Min; Wang, Ya-Fang; Wang, Jen-Ren; Hu, Mei-Hua; Chiang, Jen-Ron; Su, Ih-Jen; Chong, Pele Choi-Sing

    2013-01-01

    Enterovirus 71 (EV71) has caused several epidemics of hand, foot and mouth diseases (HFMD) in Asia. No effective EV71 vaccine is available. A randomized and open-label phase I clinical study registered with ClinicalTrials.gov #NCT01268787, aims to evaluate the safety, reactogenicity and immunogenicity of a formalin-inactivated EV71 vaccine candidate (EV71vac) at 5- and 10-µg doses. In this study we report the cross-neutralizing antibody responses from each volunteer against different subgenotypes of EV71 and CVA16. Sixty eligible healthy adults were recruited and vaccinated. Blood samples were obtained on day 0, 21 and 42 and tested against B1, B4, B5, C2, C4A, C4B and CVA16 for cross-neutralizing antibody responses. The immunogenicity of both 5- and 10- µg doses were found to be very similar. Approximately 45% of the participants had <8 pre-vaccination neutralization titers (Nt) against the B4 vaccine strain. After the first EV71vac immunization, 95% of vaccinees have >4-fold increase in Nt, but there was no further increase in Nt after the second dose. EV71vac induced very strong cross-neutralizing antibody responses in >85% of volunteers without pre-existing Nt against subgenotype B1, B5 and C4A. EV71vac elicited weak cross-neutralizing antibody responses (∼20% of participants) against a C4B and Coxsackie virus A16. Over 90% of vaccinated volunteers did not develop cross-neutralizing antibody responses (Nt<8) against a C2 strain. EV71vac can boost and significantly enhance the neutralizing antibody responses in volunteers who already had pre-vaccination antibodies against EV71 and/or CVA16. EV71vac is efficient in eliciting cross-neutralizing antibody responses against EV71 subgenotypes B1, B4, B5, and C4A, and provides the rationale for its evaluation in phase II clinical trials. ClinicalTrials.gov NCT01268787.

  7. An oligosaccharide-based HIV-1 2G12 mimotope vaccine induces carbohydrate-specific antibodies that fail to neutralize HIV-1 virions.

    PubMed

    Joyce, Joseph G; Krauss, Isaac J; Song, Hong C; Opalka, David W; Grimm, Karen M; Nahas, Deborah D; Esser, Mark T; Hrin, Renee; Feng, Meizhen; Dudkin, Vadim Y; Chastain, Michael; Shiver, John W; Danishefsky, Samuel J

    2008-10-14

    The conserved oligomannose epitope, Man(9)GlcNAc(2), recognized by the broadly neutralizing human mAb 2G12 is an attractive prophylactic vaccine candidate for the prevention of HIV-1 infection. We recently reported total chemical synthesis of a series of glycopeptides incorporating one to three copies of Man(9)GlcNAc(2) coupled to a cyclic peptide scaffold. Surface plasmon resonance studies showed that divalent and trivalent, but not monovalent, compounds were capable of binding 2G12. To test the efficacy of the divalent glycopeptide as an immunogen capable of inducing a 2G12-like neutralizing antibody response, we covalently coupled the molecule to a powerful immune-stimulating protein carrier and evaluated immunogenicity of the conjugate in two animal species. We used a differential immunoassay to demonstrate induction of high levels of carbohydrate-specific antibodies; however, these antibodies showed poor recognition of recombinant gp160 and failed to neutralize a panel of viral isolates in entry-based neutralization assays. To ascertain whether antibodies produced during natural infection could recognize the mimetics, we screened a panel of HIV-1-positive and -negative sera for binding to gp120 and the synthetic antigens. We present evidence from both direct and competitive binding assays that no significant recognition of the glycopeptides was observed, although certain sera did contain antibodies that could compete with 2G12 for binding to recombinant gp120.

  8. Microlichus americanus acariasis in saffron finches (Sicalis flaveola) with dermatitis and feather loss.

    PubMed

    Rettenmund, Christy L; Ossiboff, Robert J; McAloose, Denise; Knee, Wayne; Wade, Susan E; Paré, Jean A

    2015-05-01

    Over a 5-year period, 13 saffron finches (Sicalis flaveola) housed in mixed aviaries at the Bronx Zoo (Bronx, New York) were examined with feather loss and dermatitis, primarily affecting the nape, neck, and dorsum. Feather loss, hyperkeratosis, epidermal hyperplasia, and mixed granulocytic and mononuclear inflammation were identified in biopsies from live birds and tissue sections from postmortem specimens. In 10 of 13 cases, sections of arthropod parasites were seen histologically within feather follicles and along the surface of affected skin. Based on morphological characteristics, mites recovered from samples of formalin-fixed skin in 4 birds were identified as Microlichus americanus, an epidermoptid mite infrequently reported from wild birds and hippoboscid flies. Gross and histological lesions strongly implicate M. americanus as the cause of dermatitis affecting practically all saffron finches in the collection.

  9. External morphology of the cycliophoran dwarf male: a comparative study of Symbion pandora and S. americanus

    NASA Astrophysics Data System (ADS)

    Neves, Ricardo Cardoso; da Cunha, Marina Ribeiro; Funch, Peter; Wanninger, Andreas; Kristensen, Reinhardt Møbjerg

    2010-09-01

    Cycliophora is a recently described phylum to which only two species have been assigned so far, Symbion pandora and S. americanus. The cycliophoran life cycle is complex and alternates between asexual and sexual stages. Although not recognized as an entirely independent free-swimming stage when the phylum was first described, the dwarf male has a remarkably complex bodyplan albeit its very small size (approx. 30-40 μm in length). Aiming to increase the knowledge on the gross morphology of the cycliophoran dwarf male, specimens from S. pandora and S. americanus were analyzed by scanning electron microscopy. In both species, anterior and ventral ciliated fields, as well as paired lateral sensorial organs, were identified, thus confirming previous observations. However, new details are described herein such as the penial pouch that encloses the penis. We compare our findings on both Symbion species with the data currently available on other metazoan dwarf males.

  10. Quetiapine Inhibits Microglial Activation by Neutralizing Abnormal STIM1-Mediated Intercellular Calcium Homeostasis and Promotes Myelin Repair in a Cuprizone-Induced Mouse Model of Demyelination

    PubMed Central

    Wang, Hanzhi; Liu, Shubao; Tian, Yanping; Wu, Xiyan; He, Yangtao; Li, Chengren; Namaka, Michael; Kong, Jiming; Li, Hongli; Xiao, Lan

    2015-01-01

    Microglial activation has been considered as a crucial process in the pathogenesis of neuroinflammation and psychiatric disorders. Several antipsychotic drugs (APDs) have been shown to display inhibitory effects on microglial activation in vitro, possibly through the suppression of elevated intracellular calcium (Ca2+) concentration. However, the exact underlying mechanisms still remain elusive. In this study, we aimed to investigate the inhibitory effects of quetiapine (Que), an atypical APD, on microglial activation. We utilized a chronic cuprizone (CPZ)-induced demyelination mouse model to determine the direct effect of Que on microglial activation. Our results showed that treatment with Que significantly reduced recruitment and activation of microglia/macrophage in the lesion of corpus callosum and promoted remyelination after CPZ withdrawal. Our in vitro studies also confirmed the direct effect of Que on lipopolysaccharide (LPS)-induced activation of microglial N9 cells, whereby Que significantly inhibited the release of nitric oxide (NO) and tumor necrosis factor α (TNF-α). Moreover, we demonstrated that pretreatment with Que, neutralized the up-regulation of STIM1 induced by LPS and declined both LPS and thapsigargin (Tg)-induced store-operated Ca2+ entry (SOCE). Finally, we found that pretreatment with Que significantly reduced the translocation of nuclear factor kappa B (NF-κB) p65 subunit from cytoplasm to nuclei in LPS-activated primary microglial cells. Overall, our data suggested that Que may inhibit microglial activation by neutralization of the LPS-induced abnormal STIM1-mediated intercellular calcium homeostasis. PMID:26732345

  11. A microchannel confocal examination of arsenic speciation and distribution in Bufo americanus

    NASA Astrophysics Data System (ADS)

    Nearing, Michelle M.; Koch, Iris; Gordon, Robert A.; Reimer, Kenneth J.

    2016-05-01

    We have used confocal methods to examine the distribution and speciation of arsenic within amphibian tissue (Bufo americanus) from a contaminated mine site. The use of new microchannel technology permits a confocal, and energy-independent, examination of a given voxel within the amphibian tissue without the need for sectioning. We observe differences in arsenic concentration and speciation depending on tissue type, with the Eberth- Katschenko layer in particular containing Mn, Fe, Cu and Zn in addition to Ca and pentavalent arsenic.

  12. Cultivation of 'Candidatus Liberibacter asiaticus', 'Ca. L. africanus', and 'Ca. L. americanus' associated with huanglongbing.

    PubMed

    Sechler, A; Schuenzel, E L; Cooke, P; Donnua, S; Thaveechai, N; Postnikova, E; Stone, A L; Schneider, W L; Damsteegt, V D; Schaad, N W

    2009-05-01

    A new medium designated Liber A has been designed and used to successfully cultivate all three 'Candidatus Liberibacter spp.,' the suspect causative agents of huanglongbing (HLB) in citrus. The medium containing citrus vein extract and a growth factor sustained growth of 'Ca. Liberibacter spp.' for four or five single-colony transfers before viability declined. Colonies, positive for 'Ca. L. asiaticus' by a 16s-based rDNA real-time polymerase chain reaction (RT-PCR) assay and sequencing, were irregular-shaped, convex, and 0.1 to 0.3 mm after 3 to 4 days. Suspect 'Ca. L. asiaticus' and 'Ca. L. americanus' cells were observed in infected tissue and on agar culture by scanning electron microscopy. The cells were ovoid to rod shaped, 0.3 to 0.4 by 0.5 to 2.0 microm, often with fimbriae-like appendages. Two strains of 'Ca. L. asiaticus' and one of 'Ca. L. americanus' grown on Liber A medium were pathogenic on citrus and could be isolated from noninoculated tissues of inoculated trees and seedlings 9 and 2 months later, respectively. The identity was confirmed by RT-PCR and 16s rDNA sequencing. This is the first report of the cultivation and pathogenicity of 'Ca. L. asiaticus' and 'Ca. L. americanus' associated with symptoms of HLB.

  13. Sequence data mining in search of hookworm (Necator americanus) microRNAs.

    PubMed

    Kulkarni, Abhijeet P; Mittal, Smriti P K

    2016-09-30

    The new world hookworm, Necator americanus is a soil-transmitted nematode responsible for Necatoriasis (a type of helminthiasis) in hosts such as humans, dogs, and cats. N. americanus genome and transcriptome has been sequenced and a draft assembly analysis has been published highlighting protein coding genes and possible drug target proteins. Hookworm microRNA identification, annotations and their public release is yet to be attempted. The same is evident from lack of hookworm miRNA information in related popular public nucleotide sequence repositories such as miRBase, GenBank, WormBase etc. Therefore, in the present study we addressed these issues using EST and assembled transcript sequence information of hookworm. Using computational approaches, we identified three miRNAs precursor sequences and their mature forms. We also identified their potential targets from hookworm ESTs and transcripts, and from human transcriptome. Overall, the results indicate presence of nematode specific miRNA homologs in N. americanus and shades light on their putative targets in worm itself and the human host.

  14. Simulations of the Neutral-beam-induced Rotation, Radial Electric Field, and Flow Shearing Rate in Next-step Burning Plasmas

    SciTech Connect

    R.V. Budny

    2002-08-13

    Toroidal rotation of plasmas in present tokamaks is beneficial for increasing the stability to wall-induced MHD and appears to reduce the anomalous transport associated with micro-turbulence. This paper calculates the toroidal rotation expected from neutral-beam injection in the proposed FIRE and ITER-FEAT tokamak reactors. Self-consistent burning plasmas for these tokamaks have been constructed using the TRANSP plasma analysis code. Neutral-beam injection has been proposed for FIRE and ITER-FEAT. The neutral-beam-induced torques are computed, and assumptions for the anomalous transport of toroidal angular momentum are used to calculate the toroidal rotation profiles. The central Mach numbers are about 3-8%. The ratio of the rotation speed to the Alfvin speed is less than 1%. Assuming neoclassical poloidal rotation and force balance, the radial electric field and flow shearing rate are calculated. Peak shearing rates near the outboard edge are in the 10-100 krad/s range.

  15. An alphavirus vector overcomes the presence of neutralizing antibodies and elevated numbers of Tregs to induce immune responses in humans with advanced cancer

    PubMed Central

    Morse, Michael A.; Hobeika, Amy C.; Osada, Takuya; Berglund, Peter; Hubby, Bolyn; Negri, Sarah; Niedzwiecki, Donna; Devi, Gayathri R.; Burnett, Bruce K.; Clay, Timothy M.; Smith, Jonathan; Lyerly, H. Kim

    2010-01-01

    Therapeutic anticancer vaccines are designed to boost patients’ immune responses to tumors. One approach is to use a viral vector to deliver antigen to in situ DCs, which then activate tumor-specific T cell and antibody responses. However, vector-specific neutralizing antibodies and suppressive cell populations such as Tregs remain great challenges to the efficacy of this approach. We report here that an alphavirus vector, packaged in virus-like replicon particles (VRP) and capable of efficiently infecting DCs, could be repeatedly administered to patients with metastatic cancer expressing the tumor antigen carcinoembryonic antigen (CEA) and that it overcame high titers of neutralizing antibodies and elevated Treg levels to induce clinically relevant CEA-specific T cell and antibody responses. The CEA-specific antibodies mediated antibody-dependent cellular cytotoxicity against tumor cells from human colorectal cancer metastases. In addition, patients with CEA-specific T cell responses exhibited longer overall survival. These data suggest that VRP-based vectors can overcome the presence of neutralizing antibodies to break tolerance to self antigen and may be clinically useful for immunotherapy in the setting of tumor-induced immunosuppression. PMID:20679728

  16. Crimean-Congo Hemorrhagic Fever Virus Subunit Vaccines Induce High Levels of Neutralizing Antibodies But No Protection in STAT1 Knockout Mice.

    PubMed

    Kortekaas, Jeroen; Vloet, Rianka P M; McAuley, Alexander J; Shen, Xiaoli; Bosch, Berend Jan; de Vries, Laura; Moormann, Rob J M; Bente, Dennis A

    2015-12-01

    Crimean-Congo hemorrhagic fever virus is a tick-borne bunyavirus of the Nairovirus genus that causes hemorrhagic fever in humans with high case fatality. Here, we report the development of subunit vaccines and their efficacy in signal transducer and activator of transcription 1 (STAT1) knockout mice. Ectodomains of the structural glycoproteins Gn and Gc were produced using a Drosophila insect cell-based expression system. A single vaccination of STAT129 mice with adjuvanted Gn or Gc ectodomains induced neutralizing antibody responses, which were boosted by a second vaccination. Despite these antibody responses, mice were not protected from a CCHFV challenge infection. These results suggest that neutralizing antibodies against CCHFV do not correlate with protection of STAT1 knockout mice.

  17. Robust In Vitro and In Vivo Neutralization against Multiple High-Risk HPV Types Induced by a Thermostable Thioredoxin-L2 Vaccine.

    PubMed

    Seitz, Hanna; Ribeiro-Müller, Lis; Canali, Elena; Bolchi, Angelo; Tommasino, Massimo; Ottonello, Simone; Müller, Martin

    2015-10-01

    Current prophylactic virus-like particle (VLP) human papillomavirus (HPV) vaccines are based on the L1 major capsid protein and provide robust but virus type-restricted protection. Moreover, VLP vaccines have a high production cost, require cold-chain storage, and are thus not readily implementable in developing countries, which endure 85% of the cervical cancer-related death burden worldwide. In contrast with L1, immunization with minor capsid protein L2 elicits broad cross-neutralization, and we previously showed that insertion of a peptide spanning amino acids 20-38 of L2 into bacterial thioredoxin (Trx) greatly enhances its immunogenicity. Building on this finding, we use, here, four different neutralization assays to demonstrate that low doses of a trivalent Trx-L2 vaccine, incorporating L2(20-38) epitopes from HPV16, HPV31 and HPV51, and formulated in a human-compatible adjuvant, induce broadly protective responses. Specifically, we show that this vaccine, which uses a far-divergent archaebacterial thioredoxin as scaffold and is amenable to an easy one-step thermal purification, induces robust cross-neutralization against 12 of the 13 known oncogenic HPV types. Immune performance measured with two different in vitro neutralization assays was corroborated by the results of mouse cervico-vaginal challenge and passive transfer experiments indicating robust cross-protection also in vivo. Altogether, our results attest to the potential of Trx-L2 as a thermostable second-generation HPV vaccine particularly well suited for low-resource countries.

  18. Characteristics of HPV-Specific Antibody Responses Induced by Infection and Vaccination: Cross-Reactivity, Neutralizing Activity, Avidity and IgG Subclasses

    PubMed Central

    Scherpenisse, Mirte; Schepp, Rutger M.; Mollers, Madelief; Meijer, Chris J. L. M.; Berbers, Guy A. M.; van der Klis, Fiona R. M.

    2013-01-01

    Objectives In order to assess HPV-specific IgG characteristics, we evaluated multiple aspects of the humoral antibody response that will provide insight in the HPV humoral immune response induced by HPV infection and vaccination. Methods Cross-reactivity of HPV-specific antibodies induced by infection or vaccination was assessed with VLP16 or 18 inhibition using a VLP-based multiplex immunoassay (MIA) for HPV16, 18, 31, 33, 45, 52 and 58. HPV16/18 specific IgG1-4 subclasses and avidity were determined with the VLP-MIA in sera after HPV infection and after vaccination. Neutralizing antibodies were determined in a small subset of single-seropositive and multi-seropositive naturally derived antibodies. Results Naturally derived antibodies from single-positive sera were highly genotype-specific as homologue VLP-inhibition percentages varied between 78-94%. In multi-positive sera, cross-reactive antibodies were observed both within and between α7 and α9 species. After vaccination, cross-reactive antibodies were mainly species-specific. Avidity of vaccine-derived HPV-specific antibodies was 3 times higher than that of antibodies induced by HPV infection (p<0.0001). IgG1 and IgG3 were found to be the predominant subclasses observed after HPV infection and vaccination. In the small subset tested, the number of single-positive sera with neutralizing capacity was higher than of multi-positive sera. Conclusion Naturally derived HPV-specific antibodies from single-positive samples showed different characteristics in terms of cross-reactivity and neutralizing capacity compared with antibodies from multi-positive sera. Post-vaccination, HPV antibody avidity was approximately 3 times higher than antibody avidity induced by HPV infection. Therefore, antibody avidity might be a potential surrogate of protection. PMID:24058629

  19. Improved insights into the transcriptomes of the human hookworm Necator americanus--fundamental and biotechnological implications.

    PubMed

    Rabelo, Elida M; Hall, Ross S; Loukas, Alex; Cooper, Leanne; Hu, Min; Ranganathan, Shoba; Gasser, Robin B

    2009-01-01

    Hookworms of humans are blood-feeding parasitic nematodes of major socio-economic significance in a wide range of countries. They cause a neglected tropical disease (NTD) called "hookworm disease" (=necatoriasis and/or ancylostomiasis). Necator americanus is the most widely distributed hookworm of humans and is a leading cause of iron deficiency anaemia, which can cause physical and mental retardation and deaths in children as well as adverse maternal-foetal outcomes. Currently, there is a significant focus on the development of new approaches for the prevention and control of hookworms in humans. Technological advances are underpinning the discovery of drug and vaccine targets through insights into the molecular biology and genomics of these parasites and their relationship with the human host. In spite of the widespread socio-economic impacts of human necatoriasis, molecular datasets for N. americanus are scant, limiting progress in molecular research. The present article explores all currently available EST datasets for adult and larval stages of N. americanus using a semi-automated bioinformatic pipeline. In the current repertoire of molecules now available, some have been or are being considered as candidate vaccines against N. americanus. Among others, the most abundant sets of molecules relate to the pathogenesis-related protein (PRP) superfamily, comprising various members, such as the Ancylostoma-secreted or activation-associated proteins (ASPs) and the kunitz-type proteins, both of which are inferred to play key roles in the interplay between N. americanus and the human host. Understanding the molecular biology of these and other novel molecules discovered could have important implications for finding new ways of disrupting the pathways that they are involved in, and should facilitate the identification of new drug and vaccine targets. Also, the bioinformatic prediction of the essentiality of genes and gene products as well as molecular network

  20. Grizzly bears (Ursus arctos horribilis) and black bears (Ursus americanus) prevent trabecular bone loss during disuse (hibernation).

    PubMed

    McGee-Lawrence, Meghan E; Wojda, Samantha J; Barlow, Lindsay N; Drummer, Thomas D; Castillo, Alesha B; Kennedy, Oran; Condon, Keith W; Auger, Janene; Black, Hal L; Nelson, O Lynne; Robbins, Charles T; Donahue, Seth W

    2009-12-01

    Disuse typically causes an imbalance in bone formation and bone resorption, leading to losses of cortical and trabecular bone. In contrast, bears maintain balanced intracortical remodeling and prevent cortical bone loss during disuse (hibernation). Trabecular bone, however, is more detrimentally affected than cortical bone in other animal models of disuse. Here we investigated the effects of hibernation on bone remodeling, architectural properties, and mineral density of grizzly bear (Ursus arctos horribilis) and black bear (Ursus americanus) trabecular bone in several skeletal locations. There were no differences in bone volume fraction or tissue mineral density between hibernating and active bears or between pre- and post-hibernation bears in the ilium, distal femur, or calcaneus. Though indices of cellular activity level (mineral apposition rate, osteoid thickness) decreased, trabecular bone resorption and formation indices remained balanced in hibernating grizzly bears. These data suggest that bears prevent bone loss during disuse by maintaining a balance between bone formation and bone resorption, which consequently preserves bone structure and strength. Further investigation of bone metabolism in hibernating bears may lead to the translation of mechanisms preventing disuse-induced bone loss in bears into novel treatments for osteoporosis.

  1. Myostatin from the American lobster, Homarus americanus: Cloning and effects of molting on expression in skeletal muscles.

    PubMed

    MacLea, Kyle S; Covi, Joseph A; Kim, Hyun-Woo; Chao, Erica; Medler, Scott; Chang, Ernest S; Mykles, Donald L

    2010-12-01

    A cDNA encoding a myostatin (Mstn)-like gene from an astacuran crustacean, Homarus americanus, was cloned and characterized. Mstn inhibits skeletal muscle growth in vertebrates and may play a role in crustacean muscle as a suppressor of protein synthesis. Sequence analysis and three-dimensional modeling of the Ha-Mstn protein predicted a high degree of conservation with vertebrate and other invertebrate myostatins. Qualitative polymerase chain reaction (PCR) demonstrated ubiquitous expression of transcript in all tissues, including skeletal muscles. Quantitative PCR analysis was used to determine the effects of natural molting and eyestalk ablation (ESA) on Ha-Mstn expression in the cutter claw (CT) and crusher claw (CR) closer muscles and deep abdominal (DA) muscle. In intermolt lobsters, the Ha-Mstn mRNA level in the DA muscle was significantly lower than the mRNA levels in the CT and CR muscles. Spontaneous molting decreased Ha-Mstn mRNA during premolt, with the CR muscle, which is composed of slow-twitch (S₁) fibers, responding preferentially (82% decrease) to the atrophic signal compared to fast fibers in CT (51% decrease) and DA (69% decrease) muscles. However, acute increases in circulating ecdysteroids caused by ESA had no effect on Ha-Mstn mRNA levels in the three muscles. These data indicate that the transcription of Ha-Mstn is differentially regulated during the natural molt cycle and it is an important regulator of protein turnover in molt-induced claw muscle atrophy.

  2. Low-salinity stress in the American lobster, Homarus americanus, after chronic sublethal exposure to cadmium: Biochemical effects

    NASA Astrophysics Data System (ADS)

    Gould, E.

    1980-03-01

    Lobsters (Homarus americanus) were exposed to cadmium (6 µg l-1, 30 days) in flowing seawater, then held for 7 days in aerated “clean ” seawater at either ambient (27 ‰) or low (17 ‰) salinity. Cadmium exposure alone (ambient salinity) induced a general elevation of enzyme activity (heart, antennal gland, and muscle MDH; heart LDH and GPI), despite the probability of some clearance of cadmium from body tissues during the “clean ” seawater holding period. Low-salinity alone (non-exposed lobsters) caused a decrease of enzyme activity (AAT, LDH, GPI, PK) in most tissues examined, except for tail muscle IDH, the activity of which was increased, and MDH, which was significantly elevated above ambient controls in all tissues except heart. Most low-salinity effects were observed in tail muscle, and most cadmium effects, in heart; low-salinity effects outnumbered cadmium stress by nine to four. In heart and tail muscle of cadmium-exposed lobsters held at low salinity, each of the two stresses apparently operated to nullify the other's effects. The most prominent single biochemical response to these sublethal stresses was the elevation of MDH activity. The ratio MDH: LDH gave the clearest indication of overall relative stress.

  3. Simulation of field-induced molecular dissociation in atom-probe tomography: Identification of a neutral emission channel

    NASA Astrophysics Data System (ADS)

    Zanuttini, David; Blum, Ivan; Rigutti, Lorenzo; Vurpillot, François; Douady, Julie; Jacquet, Emmanuelle; Anglade, Pierre-Matthieu; Gervais, Benoit

    2017-06-01

    We investigate the dynamics of dicationic metal-oxide molecules under large electric-field conditions, on the basis of ab initio calculations coupled to molecular dynamics. Applied to the case of ZnO2 + in the field of atom probe tomography (APT), our simulation reveals the dissociation into three distinct exit channels. The proportions of these channels depend critically on the field strength and on the initial molecular orientation with respect to the field. For typical field strength used in APT experiments, an efficient dissociation channel leads to emission of neutral oxygen atoms, which escape detection. The calculated composition biases and their dependence on the field strength show remarkable consistency with recent APT experiments on ZnO crystals. Our work shows that bond breaking in strong static fields may lead to significant neutral atom production, and therefore to severe elemental composition biases in measurements.

  4. Antibody-Induced Conformational Changes in Herpes Simplex Virus Glycoprotein gD Reveal New Targets for Virus Neutralization

    PubMed Central

    Lazear, Eric; Whitbeck, J. Charles; Ponce-de-Leon, Manuel; Cairns, Tina M.; Willis, Sharon H.; Zuo, Yi; Krummenacher, Claude; Cohen, Gary H.

    2012-01-01

    As the receptor-binding protein of herpes simplex virus (HSV), gD plays an essential role in virus entry. In its native state, the last 56 amino acids of the ectodomain C terminus (C-term) occlude binding to its receptors, herpesvirus entry mediator (HVEM) and nectin-1. Although it is clear that movement of the C-term must occur to permit receptor binding, we believe that this conformational change is also a key event for triggering later steps leading to fusion. Specifically, gD mutants containing disulfide bonds that constrain the C-term are deficient in their ability to trigger fusion following receptor binding. In this report, we show that two newly made monoclonal antibodies (MAbs), MC2 and MC5, have virus-neutralizing activity but do not block binding of gD to either receptor. In contrast, all previously characterized neutralizing anti-gD MAbs block binding of gD to a receptor(s). Interestingly, instead of blocking receptor binding, MC2 significantly enhances the affinity of gD for both receptors. Several nonneutralizing MAbs (MC4, MC10, and MC14) also enhanced gD-receptor binding. While MC2 and MC5 recognized different epitopes on the core of gD, these nonneutralizing MAbs recognized the gD C-term. Both the neutralizing capacity and rate of neutralization of virus by MC2 are uniquely enhanced when MC2 is combined with MAb MC4, MC10, or MC14. We suggest that MC2 and MC5 prevent gD from performing a function that triggers later steps leading to fusion and that the epitope for MC2 is normally occluded by the C-term of the gD ectodomain. PMID:22130533

  5. Immunization routes in cattle impact the levels and neutralizing capacity of antibodies induced against S. aureus immune evasion proteins.

    PubMed

    Boerhout, Eveline; Vrieling, Manouk; Benedictus, Lindert; Daemen, Ineke; Ravesloot, Lars; Rutten, Victor; Nuijten, Piet; van Strijp, Jos; Koets, Ad; Eisenberg, Susanne

    2015-09-28

    Vaccines against S. aureus bovine mastitis are scarce and show limited protection only. All currently available vaccines are applied via the parenteral (usually intramuscular) route. It is unknown, however, whether this route is the most suitable to specifically increase intramammary immunity to combat S. aureus at the site of infection. Hence, in the present study, immunization via mucosal (intranasal; IN), intramuscular (triangle of the neck; IM), intramammary (IMM) and subcutaneous (suspensory ligament; SC) routes were analyzed for their effects on the quantity of the antibody responses in serum and milk as well as the neutralizing capacity of the antibodies within serum. The experimental vaccine comprised the recombinant S. aureus immune evasion proteins extracellular fibrinogen-binding protein (Efb) and the leukotoxin subunit LukM in an oil-in-water adjuvant combined with a hydrogel and alginate. The highest titer increases for both Efb and LukM specific IgG1 and IgG2 antibody levels in serum and milk were observed following SC/SC immunizations. Furthermore, the harmful effects of Efb and leukotoxin LukMF' on host-defense were neutralized by serum antibodies in a route-dependent manner. SC/SC immunization resulted in a significant increase in the neutralizing capacity of serum antibodies towards Efb and LukMF', shown by increased phagocytosis of S. aureus and increased viability of bovine leukocytes. Therefore, a SC immunization route should be considered when aiming to optimize humoral immunity against S. aureus mastitis in cattle.

  6. A novel replication-competent vaccinia vector MVTT is superior to MVA for inducing high levels of neutralizing antibody via mucosal vaccination.

    PubMed

    Huang, Xiaoxing; Lu, Bin; Yu, Wenbo; Fang, Qing; Liu, Li; Zhuang, Ke; Shen, Tingting; Wang, Haibo; Tian, Po; Zhang, Linqi; Chen, Zhiwei

    2009-01-01

    Mucosal vaccination offers great advantage for inducing protective immune response to prevent viral transmission and dissemination. Here, we report our findings of a head-to-head comparison of two viral vectors modified vaccinia Ankara (MVA) and a novel replication-competent modified vaccinia Tian Tan (MVTT) for inducing neutralizing antibodies (Nabs) via intramuscular and mucosal vaccinations in mice. MVTT is an attenuated variant of the wild-type VTT, which was historically used as a smallpox vaccine for millions of Chinese people. The spike glycoprotein (S) of SARS-CoV was used as the test antigen after the S gene was constructed in the identical genomic location of two vectors to generate vaccine candidates MVTT-S and MVA-S. Using identical doses, MVTT-S induced lower levels ( approximately 2-3-fold) of anti- SARS-CoV neutralizing antibodies (Nabs) than MVA-S through intramuscular inoculation. MVTT-S, however, was capable of inducing consistently 20-to-100-fold higher levels of Nabs than MVA-S when inoculated via either intranasal or intraoral routes. These levels of MVTT-S-induced Nab responses were substantially (approximately 10-fold) higher than that induced via the intramuscular route in the same experiments. Moreover, pre-exposure to the wild-type VTT via intranasal or intraoral route impaired the Nab response via the same routes of MVTT-S vaccination probably due to the pre-existing anti-VTT Nab response. The efficacy of intranasal or intraoral vaccination, however, was still 20-to-50-fold better than intramuscular inoculation despite the subcutaneous pre-exposure to wild-type VTT. Our data have implications for people who maintain low levels of anti-VTT Nabs after historical smallpox vaccination. MVTT is therefore an attractive live viral vector for mucosal vaccination.

  7. A multi-subunit Chlamydia vaccine inducing neutralizing antibodies and strong IFN-γ⁺ CMI responses protects against a genital infection in minipigs.

    PubMed

    Bøje, Sarah; Olsen, Anja Weinreich; Erneholm, Karin; Agerholm, Jørgen Steen; Jungersen, Gregers; Andersen, Peter; Follmann, Frank

    2016-02-01

    Chlamydia is the most widespread sexually transmitted bacterial disease and a prophylactic vaccine is highly needed. Ideally, this vaccine is required to induce a combined response of Th1 cell-mediated immune (CMI) response in concert with neutralizing antibodies. Using a novel Göttingen minipig animal model, we evaluated the immunogenicity and efficacy of a multi-subunit vaccine formulated in the strong Th1-inducing adjuvant CAF01. We evaluated a mixture of two fusion proteins (Hirep1 and CTH93) designed to promote either neutralizing antibodies or cell-mediated immunity, respectively. Hirep1 is a novel immunogen based on the variant domain (VD) 4 region from major outer membrane protein (MOMP) serovar (Sv) D, SvE and SvF, and CTH93 is a fusion molecule of three antigens (CT043, CT414 and MOMP). Pigs were immunized twice intramuscularly with either Hirep1+CTH93/CAF01, UV-inactivated Chlamydia trachomatis SvD bacteria (UV-SvD/CAF01) or CAF01. The Hirep1+CTH93/CAF01 vaccine induced a strong CMI response against the vaccine antigens and high titers of antibodies, particularly against the VD4 region of MOMP. Sera from Hirep1+CTH93/CAF01 immunized pigs neutralized C. trachomatis SvD and SvF infectivity in vitro. Both Hirep1+CTH93/CAF01 and UV-SvD/CAF01 vaccination protected pigs against a vaginal C. trachomatis SvD infection. In conclusion, the Hirep1+CTH93/CAF01 vaccine proved highly immunogenic and equally protective as UV-SvD/CAF01 showing promise for the development of a subunit vaccine against Chlamydia.

  8. A multi-subunit Chlamydia vaccine inducing neutralizing antibodies and strong IFN-γ+ CMI responses protects against a genital infection in minipigs

    PubMed Central

    Bøje, Sarah; Olsen, Anja Weinreich; Erneholm, Karin; Agerholm, Jørgen Steen; Jungersen, Gregers; Andersen, Peter; Follmann, Frank

    2016-01-01

    Chlamydia is the most widespread sexually transmitted bacterial disease and a prophylactic vaccine is highly needed. Ideally, this vaccine is required to induce a combined response of Th1 cell-mediated immune (CMI) response in concert with neutralizing antibodies. Using a novel Göttingen minipig animal model, we evaluated the immunogenicity and efficacy of a multi-subunit vaccine formulated in the strong Th1-inducing adjuvant CAF01. We evaluated a mixture of two fusion proteins (Hirep1 and CTH93) designed to promote either neutralizing antibodies or cell-mediated immunity, respectively. Hirep1 is a novel immunogen based on the variant domain (VD) 4 region from major outer membrane protein (MOMP) serovar (Sv) D, SvE and SvF, and CTH93 is a fusion molecule of three antigens (CT043, CT414 and MOMP). Pigs were immunized twice intramuscularly with either Hirep1+CTH93/CAF01, UV-inactivated Chlamydia trachomatis SvD bacteria (UV-SvD/CAF01) or CAF01. The Hirep1+CTH93/CAF01 vaccine induced a strong CMI response against the vaccine antigens and high titers of antibodies, particularly against the VD4 region of MOMP. Sera from Hirep1+CTH93/CAF01 immunized pigs neutralized C. trachomatis SvD and SvF infectivity in vitro. Both Hirep1+CTH93/CAF01 and UV-SvD/CAF01 vaccination protected pigs against a vaginal C. trachomatis SvD infection. In conclusion, the Hirep1+CTH93/CAF01 vaccine proved highly immunogenic and equally protective as UV-SvD/CAF01 showing promise for the development of a subunit vaccine against Chlamydia. PMID:26268662

  9. Collision induced dissociation of doubly-charged ions: Coulomb explosion vs. neutral loss in [Ca(urea)]{sup 2+} gas phase unimolecular reactivity via chemical dynamics simulations

    SciTech Connect

    Spezia, Riccardo; Salpin, Jean-Yves; Cimas, Alvaro; Gaigeot, Marie-Pierre; Song, Kihyung; Hase, William L.

    2012-07-01

    In this paper we report different theoretical approaches to study the gas-phase unimolecular dissociation of the doubly-charged cation [Ca(urea)]{sup 2+}, in order to rationalize recent experimental findings. Quantum mechanical plus molecular mechanical (QM/MM) direct chemical dynamics simulations were used to investigate collision induced dissociation (CID) and rotational-vibrational energy transfer for Ar{sup +} [Ca(urea)]{sup 2+} collisions. For the picosecond time-domain of the simulations, both neutral loss and Coulomb explosion reactions were found and the differences in their mechanisms elucidated. The loss of neutral urea subsequent to collision with Ar occurs via a shattering mechanism, while the formation of two singly-charged cations follows statistical (or almost statistical) dynamics. Vibrational-rotational energy transfer efficiencies obtained for trajectories that do not dissociate during the trajectory integration were used in conjunction with RRKM rate constants to approximate dissociation pathways assuming complete intramolecular vibrational energy redistribution (IVR) and statistical dynamics. This statistical limit predicts, as expected, that at long time the most stable species on the potential energy surface (PES) dominate. These results, coupled with experimental CID from which both neutral loss and Coulomb explosion products were obtained, show that the gas phase dissociation of this ion occurs by multiple mechanisms leading to different products and that reactivity on the complicated PES is dynamically complex. (authors)

  10. First record of the ant cricket Myrmecophilus (Myrmecophilina) americanus (Orthoptera: Myrmecophilidae) in Mexico.

    PubMed

    Ortega-Morales, Aldo I; Rodríguez, Quetzaly K Siller; Garza-Hernández, Javier A; Adeniran, Adebiyi A; Hernández-Triana, Luis M; Rodríguez-Pérez, Mario A

    2017-04-27

    In September 2004, the New World ant cricket, Myrmecophilus americanus Saussure, 1877, was collected in association with longhorn crazy ants, Paratrechina longicornis (Latreille. 1802), in the state of Coahuila, Mexico. We are reporting the DNA barcode using the mitochrondrial cytochrome oxidase subunit I for this first record of M. americanus in Mexico.

  11. Dengue virus specific IgY provides protection following lethal dengue virus challenge and is neutralizing in the absence of inducing antibody dependent enhancement.

    PubMed

    Fink, Ashley L; Williams, Katherine L; Harris, Eva; Alvine, Travis D; Henderson, Thomas; Schiltz, James; Nilles, Matthew L; Bradley, David S

    2017-07-01

    Dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) are severe disease manifestations that can occur following sequential infection with different dengue virus serotypes (DENV1-4). At present, there are no licensed therapies to treat DENV-induced disease. DHF and DSS are thought to be mediated by serotype cross-reactive antibodies that facilitate antibody-dependent enhancement (ADE) by binding to viral antigens and then Fcγ receptors (FcγR) on target myeloid cells. Using genetically engineered DENV-specific antibodies, it has been shown that the interaction between the Fc portion of serotype cross-reactive antibodies and FcγR is required to induce ADE. Additionally, it was demonstrated that these antibodies were as neutralizing as their non-modified variants, were incapable of inducing ADE, and were therapeutic following a lethal, antibody-enhanced infection. Therefore, we hypothesized that avian IgY, which do not interact with mammalian FcγR, would provide a novel therapy for DENV-induced disease. We demonstrate here that goose-derived anti-DENV2 IgY neutralized DENV2 and did not induce ADE in vitro. Anti-DENV2 IgY was also protective in vivo when administered 24 hours following a lethal DENV2 infection. We were also able to demonstrate via epitope mapping that both full-length and alternatively spliced anti-DENV2 IgY recognized different epitopes, including epitopes that have not been previously identified. These observations provide evidence for the potential therapeutic applications of goose-derived anti-DENV2 IgY.

  12. Correlation of protection against Japanese encephalitis virus and JE vaccine (IXIARO(®)) induced neutralizing antibody titers.

    PubMed

    Van Gessel, Yvonne; Klade, Christoph S; Putnak, Robert; Formica, Alessandra; Krasaesub, Somporn; Spruth, Martin; Cena, Bruno; Tungtaeng, Anchalee; Gettayacamin, Montip; Dewasthaly, Shailesh

    2011-08-11

    Immune sera from volunteers vaccinated in a blinded Phase 3 clinical trial with JE-VAX(®) and a new Japanese encephalitis virus (JEV) vaccine (IC51 or IXIARO), were tested for the ability to protect mice against lethal JEV challenge. Sera from IXIARO vaccinated subjects were pooled into four batches based on neutralizing antibody measured by plaque reduction neutralization test (PRNT(50) titer): high (∼200), medium (∼40-50), low (∼20) and negative (<10). Pooled sera from JE-VAX(®) vaccinated subjects (PRNT(50) titer∼55) and pooled JEV antibody negative pre-vaccination sera were used as controls. Groups of ten 6- to 7-week-old female ICR mice were injected intraperitoneally with 0.5 ml of each serum pool diluted 1:2 or 1:10, challenged approximately 18 h later with a lethal dose of either JEV strain SA14 (genotype III) or strain KE-093 (genotype I) and observed for 21 days. All mice in the non-immune serum groups developed clinical signs consistent with JEV infection or died, whereas high titer sera from both IXIARO and JE-VAX(®) sera protected 90-100% of the animals. Statistical tests showed similar protection against both JEV strains SA14 and KE-093 and protection correlated with the anti-JEV antibody titer of IXIARO sera as measured by PRNT(50). Ex vivo neutralizing antibody titers showed that almost all mice with a titer of 10 or greater were fully protected. In a separate study, analysis of geometric mean titers (GMTs) of the groups of mice vaccinated with different doses of IXIARO and challenged with JEV SA14 provided additional evidence that titers≥10 were protective.

  13. Isothermal Diagnostic Assays for Monitoring Single Nucleotide Polymorphisms in Necator americanus Associated with Benzimidazole Drug Resistance

    PubMed Central

    Rashwan, Nour; Bourguinat, Catherine; Keller, Kathy; Gunawardena, Nipul Kithsiri; de Silva, Nilanthi; Prichard, Roger

    2016-01-01

    Background Soil-transmitted helminths (STHs) are the most prevalent intestinal helminths of humans, and a major cause of morbidity in tropical and subtropical countries. The benzimidazole (BZ) drugs albendazole (ABZ) and mebendazole (MBZ) are used for treatment of human STH infections and this use is increasing dramatically with massive drug donations. Frequent and prolonged use of these drugs could lead to the emergence of anthelmintic resistance as has occurred in nematodes of livestock. Previous molecular assays for putative resistance mutations have been based mainly on PCR amplification and sequencing. However, these techniques are complicated and time consuming and not suitable for resource-constrained situations. A simple, rapid and sensitive genotyping method is required to monitor for possible developing resistance to BZ drugs. Methods To address this problem, single nucleotide polymorphism (SNP) detection assays were developed based on the Smart amplification method (SmartAmp2) to target codons 167, 198, and 200 in the β-tubulin isotype 1 gene for the hookworm Necator americanus. Findings Diagnostic assays were developed and applied to analyze hookworm samples by both SmartAmp2 and conventional sequencing methods and the results showed high concordance. Additionally, fecal samples spiked with N. americanus larvae were assessed and the results showed that the Aac polymerase used has high tolerance to inhibitors in fecal samples. Conclusion The N. americanus SmartAmp2 SNP detection assay is a new genotyping tool that is rapid, sensitive, highly specific and efficient with the potential to be used as a field tool for monitoring SNPs associated with BZ resistance. However, further validation on large numbers of field samples is required. PMID:27930648

  14. Cytotoxic T cells and neutralizing antibodies induced in rhesus monkeys by virus-like particle HIV vaccines in the absence of protection from SHIV infection.

    PubMed

    Wagner, R; Teeuwsen, V J; Deml, L; Notka, F; Haaksma, A G; Jhagjhoorsingh, S S; Niphuis, H; Wolf, H; Heeney, J L

    1998-05-25

    HIV Pr55gag has in the absence of other viral components the capacity to self assemble in budding noninfectious virus-like particles (VLP). The immunological spectrum of the HIV-1IIIB gag-derived VLP was expanded either by stable anchoring of chimeric modified gp 120 on the surface of the VLP (type 1) or by replacing sequences of the Pr55gag precursor by the V3 loop and a linear portion of the CD4 binding domain (type 2). This noninfectious antigen delivery system was evaluated for immunogenicity and efficacy in rhesus macaques without adjuvants. Intramuscular immunization with both types of VLP induced high titers of gag-specific antibodies ranging from 1/8000 to 1/510,000 for type 1 VLP and from 1/4000 to 1/16,000 for type 2 VLP. Only animals immunized with type 1 VLP developed substantial endpoint titers of env-specific antibodies (1/2000-1/32,000) with a neutralizing capacity at serum dilutions of 1/32-1/128. Gag- and env-specific cytotoxic T lymphocyte (CTL) activity was induced by both types of VLP at similar levels. Four weeks after the last immunization animals were challenged intravenously with 20 MID50 of the cell free homologous envelope simian/HIV-1IIIB chimeric challenge stock Despite HIV-1-specific neutralizing and CTL responses, all vaccinated animals became infected.

  15. Neutralization of (NK-cell-derived) B-cell activating factor by Belimumab restores sensitivity of chronic lymphoid leukemia cells to direct and Rituximab-induced NK lysis.

    PubMed

    Wild, J; Schmiedel, B J; Maurer, A; Raab, S; Prokop, L; Stevanović, S; Dörfel, D; Schneider, P; Salih, H R

    2015-08-01

    Natural killer (NK) cells are cytotoxic lymphocytes that substantially contribute to the therapeutic benefit of antitumor antibodies like Rituximab, a crucial component in the treatment of B-cell malignancies. In chronic lymphocytic leukemia (CLL), the ability of NK cells to lyse the malignant cells and to mediate antibody-dependent cellular cytotoxicity upon Fc receptor stimulation is compromised, but the underlying mechanisms are largely unclear. We report here that NK-cells activation-dependently produce the tumor necrosis factor family member 'B-cell activating factor' (BAFF) in soluble form with no detectable surface expression, also in response to Fc receptor triggering by therapeutic CD20-antibodies. BAFF in turn enhanced the metabolic activity of primary CLL cells and impaired direct and Rituximab-induced lysis of CLL cells without affecting NK reactivity per se. The neutralizing BAFF antibody Belimumab, which is approved for treatment of systemic lupus erythematosus, prevented the effects of BAFF on the metabolism of CLL cells and restored their susceptibility to direct and Rituximab-induced NK-cell killing in allogeneic and autologous experimental systems. Our findings unravel the involvement of BAFF in the resistance of CLL cells to NK-cell antitumor immunity and Rituximab treatment and point to a benefit of combinatory approaches employing BAFF-neutralizing drugs in B-cell malignancies.

  16. Secondary metabolites, cytotoxic response by neutral red retention and protective effect against H2O2 induced cytotoxicity of Sedum caespitosum.

    PubMed

    Söhretoğlu, Didem; Sabuncuoğlu, Suna

    2012-01-01

    The EtOAc, n-BuOH and H20 subextracts of the crude MeOH extract of the aerial parts of Sedum caespitosum (cav.) Dc. were screened for cytotoxicity using the neutral red assay in Chinese hamster ovary cells as well as their protective effect against H2O2 induced cytotoxicity in human red blood cells. While the extracts did not show cytotoxicity, they displayed a protective effect compared to a blank and ascorbic acid. Gallic acid (1), kaempferol 3-O-alpha-rhamnopyranoside (2), quercetin 3-O-beta-glucopyranoside (3), quercetin 3-O-alpha-rhamnopyranoside (4) and myricetin 3-O-alpha-rhamnopyranoside (5) were isolated from the EtOAc extract and identified by 1D- and 2D-NMR. The protective effects of the isolated compounds against H2O2 induced cytotoxicity in human red blood cells were evaluated and 5 was the most active.

  17. A modified fluorimetric neutral filter elution method for analyzing radiation-induced double strand break and repair.

    PubMed

    Goutham, Venkatesh H; Kamalesh, Mumbrekar D; Guruprasad, Parashiva K; Vadhiraja, Manjunath B; Satyamoorthy, Kapaettu; Rao Bola Satish, Sadashiva

    2011-07-15

    Neutral filter elution assay is one of the methods used for detection of DNA double strand breaks (DSBs). However, it is laborious, expensive, and hazardous (radiolabeled precursors for DSB detection and scintillation counter for quantification), making it a less preferred method for DSB detection. In the present study, an attempt was made to improve the existing neutral filter elution assay by making use of fluorescent dye (PicoGreen) and microfiltration assembly for eluting the fragmented DNA, thereby reducing the cost and time required for the assay. We studied the effect of dye dilution, pH conditions, and cell number as a part of method standardization. X-ray dose-response and repair kinetics in lymphocytes as well as cell lines were studied for validating the sensitivity of the assay. A linear dose-response relationship for DSBs was observed at a cell number of 4×10(5)cells, a dye dilution of 500-fold, and at pH 10. Repair kinetics revealed a time-dependent repair of DSBs up to 360 min of posttreatment, indicating its usefulness in DSB repair studies. In conclusion, the present modified method is more efficient (in terms of cell number), cost effective, less time-consuming, and less hazardous compared to the existing method. Copyright © 2011 Elsevier Inc. All rights reserved.

  18. Pre-sheath density drop induced by ion-neutral friction along plasma blobs and implications for blob velocities

    SciTech Connect

    Furno, I.; Chabloz, V.; Fasoli, A.; Loizu, J.; Theiler, C.

    2014-01-15

    The pre-sheath density drop along the magnetic field in field-aligned, radially propagating plasma blobs is investigated in the TORPEX toroidal experiment [Fasoli et al., Plasma Phys. Controlled Fusion 52, 124020 (2010)]. Using Langmuir probes precisely aligned along the magnetic field, we measure the density n{sub se} at a poloidal limiter, where blobs are connected, and the upstream density n{sub 0} at a location half way to the other end of the blobs. The pre-sheath density drop n{sub se}/n{sub 0} is then computed and its dependence upon the neutral background gas pressure is studied. At low neutral gas pressures, the pre-sheath density drop is ≈0.4, close to the value of 0.5 expected in the collisionless case. In qualitative agreement with a simple model, this value decreases with increasing gas pressure. No significant dependence of the density drop upon the radial distance into the limiter shadow is observed. The effect of reduced blob density near the limiter on the blob radial velocity is measured and compared with predictions from a blob speed-versus-size scaling law [Theiler et al., Phys. Rev. Lett. 103, 065001 (2009)].

  19. Pre-sheath density drop induced by ion-neutral friction along plasma blobs and implications for blob velocities

    NASA Astrophysics Data System (ADS)

    Furno, I.; Theiler, C.; Chabloz, V.; Fasoli, A.; Loizu, J.

    2014-01-01

    The pre-sheath density drop along the magnetic field in field-aligned, radially propagating plasma blobs is investigated in the TORPEX toroidal experiment [Fasoli et al., Plasma Phys. Controlled Fusion 52, 124020 (2010)]. Using Langmuir probes precisely aligned along the magnetic field, we measure the density nse at a poloidal limiter, where blobs are connected, and the upstream density n0 at a location half way to the other end of the blobs. The pre-sheath density drop nse/n0 is then computed and its dependence upon the neutral background gas pressure is studied. At low neutral gas pressures, the pre-sheath density drop is ≈0.4, close to the value of 0.5 expected in the collisionless case. In qualitative agreement with a simple model, this value decreases with increasing gas pressure. No significant dependence of the density drop upon the radial distance into the limiter shadow is observed. The effect of reduced blob density near the limiter on the blob radial velocity is measured and compared with predictions from a blob speed-versus-size scaling law [Theiler et al., Phys. Rev. Lett. 103, 065001 (2009)].

  20. Photon-induced Formation of Molecular Hydrogen from a Neutral Polycyclic Aromatic Hydrocarbon: 9,10-dihydroanthracene

    NASA Astrophysics Data System (ADS)

    Fu, Yi; Szczepanski, Jan; Polfer, Nick C.

    2012-01-01

    Experimental results from infrared spectroscopy and mass spectrometry provide compelling evidence that UV irradiation of the neutral polycyclic aromatic hydrocarbon (PAH) 9,10-dihydroanthracene (DHA), trapped in solid argon (12 K), results in efficient (i.e., 90% yield) conversion to anthracene and molecular hydrogen. A number of dissociation pathways involving single or double hydrogen loss are investigated computationally. Among these, two mechanisms are most credible for a one-photon dissociation process involving UV photons <5.5 eV. For the lowest-energy pathway (2.3 eV), a simultaneous lengthening of the C-H bonds of H9 and H10 gives rise to an anthracene-H2 complex. A higher-energy mechanism (3.4 eV) involves an initial lengthening of the H9 C-H bond, followed by this hydrogen "grabbing" H10, and forming H2. The high yield of this photolysis reaction suggests that similar reactions may take place for other neutral PAHs, with implications for the formation of molecular hydrogen in regions of low UV exposure, such as in dark clouds.

  1. PHOTON-INDUCED FORMATION OF MOLECULAR HYDROGEN FROM A NEUTRAL POLYCYCLIC AROMATIC HYDROCARBON: 9,10-DIHYDROANTHRACENE

    SciTech Connect

    Fu Yi; Szczepanski, Jan; Polfer, Nick C.

    2012-01-01

    Experimental results from infrared spectroscopy and mass spectrometry provide compelling evidence that UV irradiation of the neutral polycyclic aromatic hydrocarbon (PAH) 9,10-dihydroanthracene (DHA), trapped in solid argon (12 K), results in efficient (i.e., 90% yield) conversion to anthracene and molecular hydrogen. A number of dissociation pathways involving single or double hydrogen loss are investigated computationally. Among these, two mechanisms are most credible for a one-photon dissociation process involving UV photons <5.5 eV. For the lowest-energy pathway (2.3 eV), a simultaneous lengthening of the C-H bonds of H9 and H10 gives rise to an anthracene-H{sub 2} complex. A higher-energy mechanism (3.4 eV) involves an initial lengthening of the H9 C-H bond, followed by this hydrogen 'grabbing' H10, and forming H{sub 2}. The high yield of this photolysis reaction suggests that similar reactions may take place for other neutral PAHs, with implications for the formation of molecular hydrogen in regions of low UV exposure, such as in dark clouds.

  2. Studies on endoparasites of the black bear (Ursus americanus) in the southeastern United States.

    PubMed

    Crum, J M; Nettles, V F; Davidson, W R

    1978-04-01

    Examination of 53 black bears (Ursus americanus) from six states in the southeastern United States revealed at least 17 species of endoparasites, including Sarcocystis sp., Spirometra mansonoides (spargana), Macracanthorhynchus ingens, Ancylostoma caninum, Arthrocephalus lotoris, Baylisascaris transfuga, Capillaria aerophila, Capillaria putorii, Crenosoma sp., Cyathospirura sp., Dirofilaria immitis, Gnathostoma sp., Gongylonema pulchrum, microfilariae, Molineus barbatus, Physaloptera sp. and Strongyloides sp. Twelve of these represent new host records for black bear, and two are considered to be new species. Data are presented on prevalence, intensity and geographic distribution of each species. Pathologic effects were associated with infections of spargana of S. mansonoides and adults of C. aerophilia.

  3. SURGICAL CORRECTION OF BILATERAL PATELLAR LUXATION IN AN AMERICAN BLACK BEAR CUB (URSUS AMERICANUS).

    PubMed

    Bennett, Katarina R; Desmarchelier, Marion R; Bailey, Trina R

    2015-06-01

    A wild orphaned male American black bear cub ( Ursus americanus ) presented with hind limb gait abnormalities and was found to have bilateral grade 3 laterally luxating patellas. There were no other significant abnormalities detected on neurologic, radiographic, or hematologic examinations. The trochlear grooves were deepened with a chondroplasty, and the redundant soft tissues imbricated. There was a marked improvement in the bear's gait postoperatively, with an apparent full return to function. To the authors' knowledge, patellar luxation has not been reported in the Ursidae family, and the success in this case suggests that this technique may be used in large wild or captive carnivore cubs.

  4. Mediastinal teratoma in a free-ranging American black bear (Ursus americanus).

    PubMed

    Munk, Brandon A; Turner, J Chris; Keel, M Kevin

    2013-12-01

    A 2.75-yr-old female American black bear (Ursus americanus) was found emaciated and unable to rise. It was euthanized due to the perceived poor prognosis. An approximately 18 x 8 x 10-cm, multiloculated, well-demarcated mass that extended from the bifurcation of the trachea, laterally displacing the lung lobes and caudally displacing the heart. The mass contained fibrous connective tissue, cartilage, bone, nervous tissue, smooth muscle, and a variety of epithelial structures. A mediastinal teratoma was diagnosed based on the microscopic features of the neoplasm.

  5. Consumption of seeds of southwestern white pine (Pinus strobiformis) by Black Bear (Ursus americanus)

    USGS Publications Warehouse

    Mattson, David J.; Arundel, Terry A.

    2013-01-01

    We report a discovery of black bears (Ursus americanus) consuming seeds of southwestern white pine (Pinus strobiformis) on north slopes of the San Francisco Peaks near Flagstaff, Arizona, in high-elevation, mixed-species conifer forest. In one instance, a bear had obtained seeds from cones excavated from a larder horde made by a red squirrel (Tamiasciurus hudsonicus). Consumption of seeds of southwestern white pine by bears had not been previously documented. This discovery adds to the number of species of pine used by bears for food as well as the geographic range within which the behavior occurs.

  6. Fatal disseminated blastomycosis in a free-ranging American black bear (Ursus americanus).

    PubMed

    Dykstra, Jaclyn A; Rogers, Lynn L; Mansfield, Susan A; Wünschmann, Arno

    2012-11-01

    An aged, free-ranging, female, radio-collared American black bear (Ursus americanus) died after an approximately 5 month long period of weight loss. Gross necropsy findings included severe diffuse pyogranulomatous bronchopneumonia, marked granulomatous lymphadenitis of tracheobronchial lymph nodes and multiple intra-abdominal lymph nodes, chronic focal jejunal ulceration, and widespread alopecia. Histopathologic examination revealed abundant fungal organisms morphologically compatible with Blastomyces sp. within pyogranulomatous inflammatory lesions in the lungs, multiple lymph nodes, liver, kidneys, jejunum, and right adrenal gland. In addition, the haircoat had a mild infestation of chewing lice (Trichodectes pinguis euarctidos), and large numbers of rhabditid nematodes consistent with Pelodera sp. were histologically observed within hair follicles.

  7. Sinorhizobium americanus sp. nov., a new Sinorhizobium species nodulating native Acacia spp. in Mexico.

    PubMed

    Toledo, Ivonne; Lloret, Lourdes; Martínez-Romero, Esperanza

    2003-03-01

    The sinorhizobia isolated from root nodules of Acacia species native of Mexico constitute a diverse group of bacteria on the basis of their metabolic enzyme electromorphs but share restriction patterns of the PCR products of 16S rRNA genes and a common 500 kb symbiotic plasmid. They are distinguished from other Sinorhizobium species by their levels of DNA-DNA hybridization and the sequence of 16S rRNA and nifH genes. nolR gene hybridization patterns were found useful to identify sinorhizobia and characterize species. A new species, Sinorhizobium americanus, is described and the type strain is CFNEI 156 from Acacia acatlensis.

  8. Broadly Neutralizing Hemagglutinin Stalk-Specific Antibodies Induce Potent Phagocytosis of Immune Complexes by Neutrophils in an Fc-Dependent Manner.

    PubMed

    Mullarkey, Caitlin E; Bailey, Mark J; Golubeva, Diana A; Tan, Gene S; Nachbagauer, Raffael; He, Wenqian; Novakowski, Kyle E; Bowdish, Dawn M; Miller, Matthew S; Palese, Peter

    2016-10-04

    Broadly neutralizing antibodies that recognize the conserved hemagglutinin (HA) stalk have emerged as exciting new biotherapeutic tools to combat seasonal and pandemic influenza viruses. Our general understanding of the mechanisms by which stalk-specific antibodies achieve protection is rapidly evolving. It has recently been demonstrated that broadly neutralizing HA stalk-specific IgG antibodies require Fc-Fcγ receptor (FcγR) interactions for optimal protection in vivo Here we examine the neutrophil effector functions induced by stalk-specific antibodies. As the most abundant subset of blood leukocytes, neutrophils represent a critical innate effector cell population and serve an instrumental role in orchestrating downstream adaptive responses to influenza virus infection. Yet, the interplay of HA stalk-specific IgG, Fc-FcγR engagement, and neutrophils has remained largely uncharacterized. Using an in vitro assay to detect the production of reactive oxygen species (ROS), we show that human and mouse monoclonal HA stalk-specific IgG antibodies are able to induce the production of ROS by neutrophils, while HA head-specific antibodies do not. Furthermore, our results indicate that the production of ROS is dependent on Fc receptor (FcR) engagement and phagocytosis. We went on to assess the ability of monoclonal HA stalk-specific IgA antibodies to induce ROS. Consistent with our findings for monoclonal IgGs, only HA stalk-specific IgA antibodies elicited ROS production by neutrophils. This induction is dependent on the engagement of FcαR1. Taken together, our findings describe a novel FcR-dependent effector function induced by HA stalk-specific IgG and IgA antibodies, and importantly, our studies shed light on the mechanisms by which HA stalk-specific antibodies achieve protection.

  9. Broadly Neutralizing Hemagglutinin Stalk-Specific Antibodies Induce Potent Phagocytosis of Immune Complexes by Neutrophils in an Fc-Dependent Manner

    PubMed Central

    Mullarkey, Caitlin E.; Bailey, Mark J.; Golubeva, Diana A.; Tan, Gene S.; Nachbagauer, Raffael; He, Wenqian; Novakowski, Kyle E.; Bowdish, Dawn M.; Miller, Matthew S.

    2016-01-01

    ABSTRACT Broadly neutralizing antibodies that recognize the conserved hemagglutinin (HA) stalk have emerged as exciting new biotherapeutic tools to combat seasonal and pandemic influenza viruses. Our general understanding of the mechanisms by which stalk-specific antibodies achieve protection is rapidly evolving. It has recently been demonstrated that broadly neutralizing HA stalk-specific IgG antibodies require Fc-Fcγ receptor (FcγR) interactions for optimal protection in vivo. Here we examine the neutrophil effector functions induced by stalk-specific antibodies. As the most abundant subset of blood leukocytes, neutrophils represent a critical innate effector cell population and serve an instrumental role in orchestrating downstream adaptive responses to influenza virus infection. Yet, the interplay of HA stalk-specific IgG, Fc-FcγR engagement, and neutrophils has remained largely uncharacterized. Using an in vitro assay to detect the production of reactive oxygen species (ROS), we show that human and mouse monoclonal HA stalk-specific IgG antibodies are able to induce the production of ROS by neutrophils, while HA head-specific antibodies do not. Furthermore, our results indicate that the production of ROS is dependent on Fc receptor (FcR) engagement and phagocytosis. We went on to assess the ability of monoclonal HA stalk-specific IgA antibodies to induce ROS. Consistent with our findings for monoclonal IgGs, only HA stalk-specific IgA antibodies elicited ROS production by neutrophils. This induction is dependent on the engagement of FcαR1. Taken together, our findings describe a novel FcR-dependent effector function induced by HA stalk-specific IgG and IgA antibodies, and importantly, our studies shed light on the mechanisms by which HA stalk-specific antibodies achieve protection. PMID:27703076

  10. Mucosal immunization induces a higher level of lasting neutralizing antibody response in mice by a replication-competent smallpox vaccine: vaccinia Tiantan strain.

    PubMed

    Lu, Bin; Yu, Wenbo; Huang, Xiaoxing; Wang, Haibo; Liu, Li; Chen, Zhiwei

    2011-01-01

    The possible bioterrorism threat using the variola virus, the causative agent of smallpox, has promoted us to further investigate the immunogenicity profiles of existing vaccines. Here, we study for the first time the immunogenicity profile of a replication-competent smallpox vaccine (vaccinia Tiantan, VTT strain) for inducing neutralizing antibodies (Nabs) through mucosal vaccination, which is noninvasive and has a critical implication for massive vaccination programs. Four different routes of vaccination were tested in parallel including intramuscular (i.m.), intranasal (i.n.), oral (i.o.), and subcutaneous (s.c.) inoculations in mice. We found that one time vaccination with an optimal dose of VTT was able to induce anti-VTT Nabs via each of the four routes. Higher levels of antiviral Nabs, however, were induced via the i.n. and i.o. inoculations when compared with the i.m. and s.c. routes. Moreover, the i.n. and i.o. vaccinations also induced higher sustained levels of Nabs overtime, which conferred better protections against homologous or alternating mucosal routes of viral challenges six months post vaccination. The VTT-induced immunity via all four routes, however, was partially effective against the intramuscular viral challenge. Our data have implications for understanding the potential application of mucosal smallpox vaccination and for developing VTT-based vaccines to overcome preexisting antivaccinia immunity.

  11. Mucosal Immunization Induces a Higher Level of Lasting Neutralizing Antibody Response in Mice by a Replication-Competent Smallpox Vaccine: Vaccinia Tiantan Strain

    PubMed Central

    Lu, Bin; Yu, Wenbo; Huang, Xiaoxing; Wang, Haibo; Liu, Li; Chen, Zhiwei

    2011-01-01

    The possible bioterrorism threat using the variola virus, the causative agent of smallpox, has promoted us to further investigate the immunogenicity profiles of existing vaccines. Here, we study for the first time the immunogenicity profile of a replication-competent smallpox vaccine (vaccinia Tiantan, VTT strain) for inducing neutralizing antibodies (Nabs) through mucosal vaccination, which is noninvasive and has a critical implication for massive vaccination programs. Four different routes of vaccination were tested in parallel including intramuscular (i.m.), intranasal (i.n.), oral (i.o.), and subcutaneous (s.c.) inoculations in mice. We found that one time vaccination with an optimal dose of VTT was able to induce anti-VTT Nabs via each of the four routes. Higher levels of antiviral Nabs, however, were induced via the i.n. and i.o. inoculations when compared with the i.m. and s.c. routes. Moreover, the i.n. and i.o. vaccinations also induced higher sustained levels of Nabs overtime, which conferred better protections against homologous or alternating mucosal routes of viral challenges six months post vaccination. The VTT-induced immunity via all four routes, however, was partially effective against the intramuscular viral challenge. Our data have implications for understanding the potential application of mucosal smallpox vaccination and for developing VTT-based vaccines to overcome preexisting antivaccinia immunity. PMID:21765641

  12. The peptide hormone pQDLDHVFLRFamide (crustacean myosuppressin) modulates the Homarus americanus cardiac neuromuscular system at multiple sites.

    PubMed

    Stevens, J S; Cashman, C R; Smith, C M; Beale, K M; Towle, D W; Christie, A E; Dickinson, P S

    2009-12-01

    pQDLDHVFLRFamide is a highly conserved crustacean neuropeptide with a structure that places it within the myosuppressin subfamily of the FMRFamide-like peptides. Despite its apparent ubiquitous conservation in decapod crustaceans, the paracrine and/or endocrine roles played by pQDLDHVFLRFamide remain largely unknown. We have examined the actions of this peptide on the cardiac neuromuscular system of the American lobster Homarus americanus using four preparations: the intact animal, the heart in vitro, the isolated cardiac ganglion (CG), and a stimulated heart muscle preparation. In the intact animal, injection of myosuppressin caused a decrease in heartbeat frequency. Perfusion of the in vitro heart with pQDLDHVFLRFamide elicited a decrease in the frequency and an increase in the amplitude of heart contractions. In the isolated CG, myosuppressin induced a hyperpolarization of the resting membrane potential of cardiac motor neurons and a decrease in the cycle frequency of their bursting. In the stimulated heart muscle preparation, pQDLDHVFLRFamide increased the amplitude of the induced contractions, suggesting that myosuppressin modulates not only the CG, but also peripheral sites. For at least the in vitro heart and the isolated CG, the effects of myosuppressin were dose-dependent (10(-9) to 10(-6) mol l(-1) tested), with threshold concentrations (10(-8)-10(-7) mol l(-1)) consistent with the peptide serving as a circulating hormone. Although cycle frequency, a parameter directly determined by the CG, consistently decreased when pQDLDHVFLRFamide was applied to all preparation types, the magnitudes of this decrease differed, suggesting the possibility that, because myosuppressin modulates the CG and the periphery, it also alters peripheral feedback to the CG.

  13. The peptide hormone pQDLDHVFLRFamide (crustacean myosuppressin) modulates the Homarus americanus cardiac neuromuscular system at multiple sites

    PubMed Central

    Stevens, J. S.; Cashman, C. R.; Smith, C. M.; Beale, K. M.; Towle, D. W.; Christie, A. E.; Dickinson, P. S.

    2009-01-01

    pQDLDHVFLRFamide is a highly conserved crustacean neuropeptide with a structure that places it within the myosuppressin subfamily of the FMRFamide-like peptides. Despite its apparent ubiquitous conservation in decapod crustaceans, the paracrine and/or endocrine roles played by pQDLDHVFLRFamide remain largely unknown. We have examined the actions of this peptide on the cardiac neuromuscular system of the American lobster Homarus americanus using four preparations: the intact animal, the heart in vitro, the isolated cardiac ganglion (CG), and a stimulated heart muscle preparation. In the intact animal, injection of myosuppressin caused a decrease in heartbeat frequency. Perfusion of the in vitro heart with pQDLDHVFLRFamide elicited a decrease in the frequency and an increase in the amplitude of heart contractions. In the isolated CG, myosuppressin induced a hyperpolarization of the resting membrane potential of cardiac motor neurons and a decrease in the cycle frequency of their bursting. In the stimulated heart muscle preparation, pQDLDHVFLRFamide increased the amplitude of the induced contractions, suggesting that myosuppressin modulates not only the CG, but also peripheral sites. For at least the in vitro heart and the isolated CG, the effects of myosuppressin were dose-dependent (10−9 to 10−6 mol l−1 tested), with threshold concentrations (10−8−10−7 mol l−1) consistent with the peptide serving as a circulating hormone. Although cycle frequency, a parameter directly determined by the CG, consistently decreased when pQDLDHVFLRFamide was applied to all preparation types, the magnitudes of this decrease differed, suggesting the possibility that, because myosuppressin modulates the CG and the periphery, it also alters peripheral feedback to the CG. PMID:19946074

  14. Neutral atom traps.

    SciTech Connect

    Pack, Michael Vern

    2008-12-01

    This report describes progress in designing a neutral atom trap capable of trapping sub millikelvin atom in a magnetic trap and shuttling the atoms across the atom chip from a collection area to an optical cavity. The numerical simulation and atom chip design are discussed. Also, discussed are preliminary calculations of quantum noise sources in Kerr nonlinear optics measurements based on electromagnetically induced transparency. These types of measurements may be important for quantum nondemolition measurements at the few photon limit.

  15. Neutral escape at Mars induced by the precipitation of high-energy protons and hydrogen atoms of the solar wind origin

    NASA Astrophysics Data System (ADS)

    Shematovich, Valery I.

    2017-04-01

    One of the first surprises of the NASA MAVEN mission was the observation by the SWIA instrument of a tenuous population of protons with solar wind energies travelling anti-sunward near periapsis, at altitudes of 150-250 km (Halekas et al., 2015). While the penetration of solar wind protons to low altitude is not completely unexpected given previous Mars Express results, this population maintains exactly the same velocity as the solar wind observed. From previous studies it was known that some fraction of the solar wind can interact with the extended corona of Mars. By charge exchange with the neutral particles in this corona, some fraction of the incoming solar wind protons can gain an electron and become an energetic neutral hydrogen atom. Once neutral, these particles penetrate through the Martian induced magnetosphere with ease, with free access to the collisional atmosphere/ionosphere. The origin, kinetics and transport of the suprathermal O atoms in the transition region (from thermosphere to exosphere) of the Martian upper atmosphere due to the precipitation of the high-energy protons and hydrogen atoms are discussed. Kinetic energy distribution functions of suprathermal and superthermal (ENA) oxygen atoms formed in the Martian upper atmosphere were calculated using the kinetic Monte Carlo model (Shematovich et al., 2011, Shematovich, 2013) of the high-energy proton and hydrogen atom precipitation into the atmosphere. These functions allowed us: (a) to estimate the non-thermal escape rates of neutral oxygen from the Martian upper atmosphere, and (b) to compare with available MAVEN measurements of oxygen corona. Induced by precipitation the escape of hot oxygen atoms may become dominant under conditions of extreme solar events - solar flares and coronal mass ejections, - as it was shown by recent observations of the NASA MAVEN spacecraft (Jakosky et al., 2015). This work is supported by the RFBR project and by the Basic Research Program of the Praesidium of

  16. Modified sugar beet pectin induces apoptosis of colon cancer cells via an interaction with the neutral sugar side-chains.

    PubMed

    Maxwell, Ellen G; Colquhoun, Ian J; Chau, Hoa K; Hotchkiss, Arland T; Waldron, Keith W; Morris, Victor J; Belshaw, Nigel J

    2016-01-20

    Pectins extracted from a variety of sources and modified with heat and/or pH have previously been shown to exhibit activity towards several cancer cell lines. However, the structural basis for the anti-cancer activity of modified pectin requires clarification. Sugar beet and citrus pectin extracts have been compared. Pectin extracted from sugar beet pulp only weakly affected the viability of colon cancer cells. Alkali treatment increased the anti-cancer effect of sugar beet pectin via an induction of apoptosis. Alkali treatment decreased the degree of esterification (DE) and increased the ratio of rhamnogalacturonan I (RGI) to homogalacturonan. Low DE per se did not play a significant role in the anti-cancer activity. However, the enzymatic removal of galactose and, to a lesser extent, arabinose from the pectin decreased the effect on cancer cells indicating that the neutral sugar-containing RGI regions are important for pectin bioactivity.

  17. Insertion of primary syncytium-inducing (SI) and non-SI envelope V3 loops in human immunodeficiency virus type 1 (HIV-1) LAI reduces neutralization sensitivity to autologous, but not heterologous, HIV-1 antibodies.

    PubMed Central

    Hogervorst, E; de Jong, J; van Wijk, A; Bakker, M; Valk, M; Nara, P; Goudsmit, J

    1995-01-01

    The aim of the study was to investigate the influence of V3 loops from naturally occurring viruses on the neutralization sensitivity of a molecularly cloned virus. A selection of well-defined syncytium-inducing (SI) and non-SI V3 loops of a single human immunodeficiency virus type 1-infected individual (H594) and the V3 regions of two SI laboratory strains were inserted in an infectious molecular clone of human immunodeficiency type 1 LAI. Neutralization was performed with a heterologous serum pool and autologous patient serum, using the virus reduction neutralization assay and peripheral blood lymphocytes as target cells. High sensitivity of the chimeric viruses containing the laboratory strain V3 regions to neutralization by H594 sequential sera as well as the heterologous serum pool was found. A statistically significant correlation between the sensitivities of these viruses was seen. In contrast, insertion of the primary isolate NSI and SI envelope V3 loops significantly reduced the neutralization by autologous serum but not by the heterologous serum pool. No correlation was found between the neutralization of the viruses with laboratory strain-derived V3 regions and the viruses with primary isolate V3 domains. We conclude that heterologous antibodies are able to neutralize infectious molecular clones with V3 loops of both SI and NSI viruses, regardless of whether they originated from laboratory strains or primary isolates. However, serum of patient H594 discriminated between the two types of viruses and showed reduced neutralization of the viruses with the autologous NSI and SI primary isolate V3 loops. These results indicated that the neutralization sensitivity of the viruses depended on the capacity of the V3 region to influence the conformation of the virus envelope. These V3-dependent conformational changes partially explain the neutralization sensitivity of laboratory strains and the relative neutralization resistance of primary isolates. PMID:7666535

  18. Selective tonicity-induced expression of the neutral amino-acid transporter SNAT2 in oligodendrocytes in rat brain following systemic hypertonicity.

    PubMed

    Maallem, S; Mutin, M; González-González, I M; Zafra, F; Tappaz, M L

    2008-04-22

    Sodium-coupled neutral amino-acid transporter member 2 (SNAT2) belongs to the family of neutral amino-acid transporters. SNAT2 is encoded by the gene Slc38a2, whose expression was reported to increase in vitro in fibroblasts, endothelial and renal cells exposed to a hypertonic medium. SNAT2 tonicity-induced expression brings about cellular accumulation of amino-acid, which contributes to osmoadaptation to hypertonicity. Since brain osmoadaptation is observed in relationship to neurological disorders resulting from pathological osmotic imbalances in blood plasma, we have investigated, through immunocytochemistry, SNAT2 expression in brain of rats subjected to systemic hypertonicity. Following prolonged systemic hypertonicity (24 h), small, strongly immunolabeled elements were observed that were not present in sham-treated animals. They were evenly distributed in the gray matter, with a lower density in the forebrain and a higher density in the brain stem. However the highest density by far was observed in white matter, where they were frequently aligned in chain-like rows. These observations suggested an oligodendrocyte location that was further established by double immunofluorescent labeling, using the oligodendrocyte phenotypic markers 2'-3'-cyclic nucleotide 3'phosphodiesterase and carbonic anhydrase II. SNAT2-positive elements were found associated with oligodendrocyte cell bodies, while oligodendrocyte processes were devoid of labeling. A quantitative analysis performed in the cerebral cortex indicated that virtually all SNAT2-positive elements were associated with oligodendrocyte cell bodies and conversely that the overwhelming majority of oligodendrocytes showed SNAT2 immunolabeling. The tonicity-induced expression of SNAT2 was not observed following acute systemic hypertonicity (6 h). Our results suggest that the osmoadaptation of brain oligodendrocytes to hypertonicity relies upon amino-acid accumulation through the tonicity-induced expression of SNAT2

  19. Elicitation of Neutralizing Antibodies Directed against CD4-Induced Epitope(s) Using a CD4 Mimetic Cross-Linked to a HIV-1 Envelope Glycoprotein

    PubMed Central

    Dey, Antu K.; Burke, Brian; Sun, Yide; Sirokman, Klara; Nandi, Avishek; Hartog, Karin; Lian, Ying; Geonnotti, Anthony R.; Montefiori, David; Franti, Michael; Martin, Grégoire; Carfi, Andrea; Kessler, Pascal; Martin, Loïc; Srivastava, Indresh K.; Barnett, Susan W.

    2012-01-01

    The identification of HIV-1 envelope glycoprotein (Env) structures that can generate broadly neutralizing antibodies (BNAbs) is pivotal to the development of a successful vaccine against HIV-1 aimed at eliciting effective humoral immune responses. To that end, the production of novel Env structure(s) that might induce BNAbs by presentation of conserved epitopes, which are otherwise occluded, is critical. Here, we focus on a structure that stabilizes Env in a conformation representative of its primary (CD4) receptor-bound state, thereby exposing highly conserved “CD4 induced” (CD4i) epitope(s) known to be important for co-receptor binding and subsequent virus infection. A CD4-mimetic miniprotein, miniCD4 (M64U1-SH), was produced and covalently complexed to recombinant, trimeric gp140 envelope glycoprotein (gp140) using site-specific disulfide linkages. The resulting gp140-miniCD4 (gp140-S-S-M64U1) complex was recognized by CD4i antibodies and the HIV-1 co-receptor, CCR5. The gp140-miniCD4 complex elicited the highest titers of CD4i binding antibodies as well as enhanced neutralizing antibodies against Tier 1 viruses as compared to gp140 protein alone following immunization of rabbits. Neutralization against HIV-27312/V434M and additional serum mapping confirm the specific elicitation of antibodies directed to the CD4i epitope(s). These results demonstrate the utility of structure-based approach in improving immunogenic response against specific region, such as the CD4i epitope(s) here, and its potential role in vaccine application. PMID:22291921

  20. An immunogen containing four tandem 10E8 epitope repeats with exposed key residues induces antibodies that neutralize HIV-1 and activates an ADCC reporter gene.

    PubMed

    Sun, Zhiwu; Zhu, Yun; Wang, Qian; Ye, Ling; Dai, Yanyan; Su, Shan; Yu, Fei; Ying, Tianlei; Yang, Chinglai; Jiang, Shibo; Lu, Lu

    2016-06-22

    After three decades of intensive research efforts, an effective vaccine against HIV-1 remains to be developed. Several broadly neutralizing antibodies to HIV-1, such as 10E8, recognize the membrane proximal external region (MPER) of the HIV-1 gp41 protein. Thus, the MPER is considered to be a very important target for vaccine design. However, the MPER segment has very weak immunogenicity and tends to insert its epitope residues into the cell membrane, thereby avoiding antibody binding. To address this complication in vaccine development, we herein designed a peptide, designated 10E8-4P, containing four copies of the 10E8 epitope as an immunogen. As predicted by structural simulation, 10E8-4P exhibits a well-arranged tandem helical conformation, with the key residues in the 10E8 epitope oriented at different angles, thus suggesting that some of these key residues may be exposed outside of the lipid membrane. Compared with a peptide containing a single 10E8 epitope (10E8-1P), 10E8-4P not only exhibited better antigenicity but also elicited neutralizing antibody response against HIV-1 pseudoviruses, whereas 10E8-1P could not induce detectable neutralizing antibody response. Importantly, antibodies elicited by 10E8-4P also possessed a strong ability to activate an antibody-dependent cell-mediated cytotoxicity (ADCC) reporter gene, thus suggesting that they may have ADCC activity. Therefore, this strategy shows promise for further optimization and application in future HIV-1 vaccine design.

  1. Seasonal and daily plasma corticosterone rhythms in American toads, Bufo americanus

    SciTech Connect

    Pancak, M.K.; Taylor, D.H.

    1983-06-01

    Concentrations of corticosterone were measured in the plasma of American toads, Bufo americanus, on a seasonal basis using a radioimmunoassay technique. Two populations of toads, maintained under different light conditions, were monitored to observe the effects of photoperiod on the seasonal rhythm of plasma corticosterone. Under a natural photoperiod toads demonstrated a rhythm consisting of a spring peak and a fall peak in corticosterone concentration. Toads maintained under a 12L:12D photoperiod all year round demonstrated a similar rhythm with peaks in the spring and fall. This suggests that an endogenous (circannual) rhythm of corticosterone may be playing an important role in the seasonal change of overt behavior and physiology of Bufo americanus. A daily rhythm of corticosterone was also detected in toads when blood samples were taken every 4 hr. When compared to a previously published circadian rhythm study of locomotor activity, the surge in corticosterone concentration for the day occurred at 1730 just prior to the peak in locomotor activity.

  2. Chaoborus americanus predation on various zooplankters; Functional response and behavioral observations.

    PubMed

    Vinyard, Gary L; Menger, R A

    1980-04-01

    Predation rates for Chaoborus americanus on different types of noncyclomorphic zooplankton prey were determined in the laboratory as a function of both prey species and density. The sequence of events leading to consumption of prey was also observed, and probabilities determined for the various components.Predation rates generally reached an upper limit as the density of prey increased to high levels. Larger prey were always less vulnerable to Chaoborus predation. Among the species of zooplankton prey examined Holopedium gibberum suffered the lowest rates of predation, while those of the other plankters (Daphnia pulex, D. rosea, Moina hutchinsoni, Diaptomus birgei, D. leptopus, Cyclops vernalis) were approximately equivalent for any given size category.Observation of the predatory sequence indicates that for all the plankters except H. gibberum there were relatively high rates of evasion of attack by C. americanus and low rates of escape after capture. For H. gibberum the situation was reversed, and escape after capture was particularly important in determining the predation rate.The gelatinous envelope of Holopedium gibberum seems to operate as an effective anti-predator device which reduces the likelyhood of retention by Chaoborus after contact has been made.

  3. Sarcocystis sp. in mountain goats (Oreamnos americanus) in Washington: prevalence and search for the definitive host.

    PubMed

    Foreyt, W J

    1989-10-01

    During October and November 1986, Sarcocystis sp. was detected in 24 of 56 (43%) tongues from hunter-killed mountain goats (Oreamnos americanus) in Washington (USA). Sarcocysts had an unusual sessile polyp-shaped branched wall. Mean size of 154 sarcocysts was 71.3 x 37.8 microns (range, 20 to 248 x 10 to 120 microns), and the mean intensity was 2.3 (range, 1 to 28). In an attempt to identify the definitive host, infected tongues were fed to four coyotes (Canis latrans), eight domestic dogs, four domestic cats, three bears (Ursus americanus), two raccoons (Procyon lotor), two martens (Martes americana), two fishers (Martes pennanti), three skunks (Mephitis mephitis), five mink (Mustela vison), five ferrets (Mustela putorius), one pigtail macaque (Macaca nemestrina), two red-tailed hawks (Buteo jamaicensis) and one great horned owl (Bubo virginianus). Oocysts or sporocysts were not detected in the feces of any host for less than or equal to 20 days after ingestion of the infected meat. The definitive host for Sarcocystis sp. in mountain goats was not identified.

  4. Developmental Effects of Ocean Acidification Conditions and Elevated Temperature on Homarus Americanus Larvae

    NASA Astrophysics Data System (ADS)

    Mcveigh, H.; Waller, J. D.

    2016-02-01

    The Gulf of Maine is experiencing a rapid warming in sea surface temperature and a marked decrease in pH. This study aimed to quantify the impact of elevated temperature and acidification on the larval development of the iconic American lobster (Homarus americanus). Experimental conditions were reflective of current and IPCC predicted levels of temperature and pCO2 to be reached by the end of the century. Larvae were measured for growth (carapace length), development time, and survivorship over the larval duration. Treatments of elevated temperatures experienced decreased development time across the larval stages of H. americanus. Consequently mortality increased at a significantly higher rate under elevated temperature. An increase in larval mortality may decrease recruitment to the commercial fishery, thus impacting the most valuable single species in the state of Maine. Furthermore, experimental pCO2 treatments yielded a significantly decreased development time between larval stages II and III, yet did not have a significant impact on carapace length or mortality. This study indicates that warmer temperatures may have a greater influence than decreased pH on larval development and survival. Determining how this species may respond to changing climactic conditions will better inform the sustainability efforts of such a critical marine fishery.

  5. A novel isothermal microcalorimetry tool to assess drug effects on Ancylostoma ceylanicum and Necator americanus.

    PubMed

    Flores, Dayana; Panic, Gordana; Braissant, Olivier; Keiser, Jennifer

    2016-01-01

    Soil-transmitted helminths, which affect the poorest communities, worldwide cause a range of symptoms and morbidity, yet few treatment options are available and drug resistance is a concern. To improve and accelerate anthelminthic drug discovery, novel drug screening tools such as isothermal microcalorimetry (IMC) have been tested with great potential. In this study, we used a novel microcalorimeter, the calScreener™, to study the viability on the hookworms Necator americanus and Ancylostoma ceylanicum as well as the whipworm Trichuris muris. Significant heat flow signals could be obtained with already one adult worm per channel for all three species. High-amplitude oscillations were observed for the hookworms; however, adult T. muris showed a twofold heat flow decrease during the first 24 h. Antinematodal effects of ivermectin and levamisole at 1, 10, and 100 μg/ml were evaluated on adult N. americanus and A. ceylanicum. Levamisole-treated hookworms showed a decline in heat flow and oscillation amplitude in a dose-response manner. Heat flow for ivermectin-treated hookworms increased proportionally with increased concentrations of ivermectin, though the wavelet analysis showed an opposite trend as observed by flatter wavelets. In conclusion, the calScreener™ is an excellent tool to study drug effects on intestinal hookworms at the adult worm stage as it offers a lower detection limit than other IMC devices and the possibility to monitor worm viability online.

  6. Cloning and expression study of the lobster (Homarus americanus) vitellogenin: Conservation in gene structure among decapods.

    PubMed

    Tiu, Shirley Hiu Kwan; Hui, Ho-Lam; Tsukimura, Brian; Tobe, Stephen S; He, Jian-Guo; Chan, Siu-Ming

    2009-01-01

    This study reports the molecular characterization of the vitellogenin (Vg) of the lobster, Homarus americanus. Based on the annual collection of female lobsters, vitellogenesis commences in early March and continues through to September of each year. Using an antibody to vitellin of the lobster, H. americanus, several immunoreactive ovarian proteins were initially identified by Western blot analysis. The 80kDa protein contained the amino acid sequence APWGGNTPRC, identified subsequently by cDNA cloning to be identical to the lobster Vg. In common with the shrimp Metapenaeus ensis and crab Charybdis feriatus, the lobster HaVg1 gene comprises 14 introns and 15 exons. The deduced HaVg1 precursor is most similar to the Vg of the crayfish Cherax quadricarinatus (57%), followed by M. ensis (40-43% identity) and C. feriatus (38%). The results from genomic and RT-PCR cloning also confirmed the presence of multiple Vg genes in lobster. At early reproductive stages, the hepatopancreas HaVg1 transcript levels are low but increased to a maximum in animals with mature oocytes. The ovary, however, also expressed low levels of HaVg1. Using in vitro explant culture, treatment of hepatopancreas fragments with farnesoic acid or 20-hydroxyecdysone resulted in a significant stimulation in HaVg1 expression. From this study, it appears that Vg gene organization and expression pattern in decapods is highly conserved. Similar endocrine mechanisms may govern the process of vitellogenesis across the decapods.

  7. The epidemiology and population biology of Necator americanus infection in a rural community in Zimbabwe.

    PubMed

    Bradley, M; Chandiwana, S K; Bundy, D A; Medley, G F

    1992-01-01

    Baseline data from an epidemiological study of hookworm infection in a rural community in Zimbabwe are presented. The infection status of an age-stratified sample of the community was assessed using anthelmintic expulsion techniques. Necator americanus was the only helminth parasite found to be present. The age-prevalence and intensity profiles rose asymptotically to an adult prevalence of about 80% and adult mean burden of 7.7 worms per host. The overall mean burden was 4.8 worms per host. The frequency distribution of N. americanus was overdispersed and well described by the negative binomial distribution with a value for the aggregation parameter, k, of 0.346. Separate estimates of k were lower in males and older hosts. The distribution patterns were difficult to reconcile with any simple process of age-dependent acquisition of an effective immune response. A significant negative correlation was recorded between per caput fecundity and worm burden, providing evidence for a density-dependent regulation of female worm fecundity. The basic reproductive rate (R0 congruent to 2) was found to be similar to estimates from other geographical areas.

  8. Phytoremediation of two types of sediment contaminated with Zn by Schoenoplectus americanus.

    PubMed

    Arreghini, Silvana; de Cabo, Laura; Fabrizio de Iorio, Alicia

    2006-01-01

    The effect of different sediments on growth, Zn uptake, Zn plant distribution, and morphometric variables of Schoenoplectus americanus were investigated under controlled conditions. Two types of sediments were assayed: from a large natural levee (LS) and alluvial sediments (AS), the former with lower organic matter (OM) and nutrients content than AS, without and with added Zn (2500 microg Zn/g air-dry sediment). Zinc partition in sediment was determined. Increases in water conductivity and Zn concentrations in water and sediments were observed in artificially contaminated treatments. Plants showed a lower above ground growth rate, height, and width of shoots, and a higher Zn concentration in shoots and rhizomes. In the contaminated treatments, AS treatment showed lower Zn concentration in water and higher Zn concentration in sediments (total, exchangeable, and OM fractions) than LS treatment, due to Zn displacement from floodwater to sediments. The presence of a high level of OM and nutrients also increased aboveground biomass growth, whereas it decreased Zn concentration in shoots. Although the translocation factor increased with Zn addition, it was lower in AS treatment Sediments of AS treatments are a suitable environment for growth of S. americanus, which partially compensates the toxic effects of Zn. Our results provide an encouraging basis for planning larger scale experiments to test the role of OM and nutrients in improving phytoremediation.

  9. A study on three-dimensional structures of the ionospheric dynamo currents induced by the neutral winds simulated by the Kyushu-GCM

    NASA Astrophysics Data System (ADS)

    Kawano-Sasaki, Keiko; Miyahara, Saburo

    2008-08-01

    Three-dimensional structures of the ionospheric dynamo currents are examined using the neutral winds in a general circulation model of the middle atmosphere at Kyushu University. A quasi-three-dimensional ionospheric dynamo model is constructed assuming an infinite parallel conductivity in the ionosphere. This model is able to simulate both the equatorial electrojet and the global Sq current system successfully. The simulated results reveal that the equatorial electrojet is confined in quite narrow latitudes around the equator accompanied with meridional current circulations and satisfies a non-divergent structure mainly within the E region. A vertically stratified double layered structure is seen in the east-west current density near the focus latitude of the global Sq current system. It is shown that the stratified structure mainly consists of the east-west Hall current associated with the eastward wind of zonal wavenumbers 1 and 2 in the lower altitudes and the westward wind of zonal wavenumber 2 in the upper altitudes. The day-to-day variation of the neutral winds can significantly vary the induced ionospheric dynamo current system, which is recognized as changes of the focus latitude and/or the maximum value of the equatorial electrojet.

  10. Peste des petits ruminants virus-like particles induce both complete virus-specific antibodies and virus neutralizing antibodies in mice.

    PubMed

    Liu, Fuxiao; Wu, Xiaodong; Zou, Yanli; Li, Lin; Wang, Zhiliang

    2015-03-01

    Peste des petits ruminants virus (PPRV), an etiological agent of peste des petits ruminants (PPR), is classified into the genus Morbillivirus in the family Paramyxoviridae. In a previous study, a recombinant baculovirus has been constructed to co-express the PPRV matrix (M), haemagglutinin (H) and nucleocapsid (N) proteins in insect cells, causing budding of PPR virus-like particles (VLPs) from insect cell membranes by viewing of ultrathin section with a transmission electron microscope. In this follow-up study, these PPR VLPs were purified by sucrose density gradient centrifugation for immunizing mice twice. Three weeks post-primary immunization and 2 weeks post-secondary immunization, all serum samples were obtained and subsequently subjected to indirect ELISA detection on complete virus-specific antibodies. In addition, all serum samples, which were collected 2 weeks post-secondary immunization, were used for virus neutralization test on PPRV neutralizing antibodies. The results showed that the purified PPR VLPs induced both types of antibodies mentioned above in mice, indicating a given potential of VLP-based vaccine candidate against PPR. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. Recombinant retrovirus-derived virus-like particle-based vaccines induce hepatitis C virus-specific cellular and neutralizing immune responses in mice.

    PubMed

    Huret, Christophe; Desjardins, Delphine; Miyalou, Mathilde; Levacher, Béatrice; Amadoudji Zin, Martin; Bonduelle, Olivia; Combadière, Béhazine; Dalba, Charlotte; Klatzmann, David; Bellier, Bertrand

    2013-03-01

    While the immunological correlates of hepatitis C virus (HCV)-specific immunity are not well understood, it is now admitted that an effective vaccine against HCV will need to induce both cellular and humoral immune responses and address viral heterogeneity to prevent immune escape. We developed a vaccine platform specifically aimed at inducing such responses against HCV antigens displayed by recombinant retrovirus-based virus-like particles (VLPs) made of Gag of murine leukemia virus. Both ex vivo produced VLPs and plasmid DNA encoding VLPs can be used as vaccines. Here, we report that immunizations with plasmid DNA forming VLPs pseudotyped with HCV E1 and E2 envelope glycoproteins (HCV-specific plasmo-retroVLPs) induce strong T-cell-mediated immune responses that can be optimized by using proper DNA delivery methods and/or genetic adjuvants. Additionally, multigenotype or multi-specific T-cell responses were observed after immunization with plasmids that encode VLPs pseudotyped with E1E2 derived from numerous viral genotypes and/or displaying NS3 antigen in capsid proteins. While homologous prime-boost immunizations with HCV-specific plasmo-retroVLPs or ex vivo produced VLPs induce a low level of specific antibody responses, optimal combination of plasmo-retroVLPs and VLPs was identified for inducing HCV-specific T-cell and B-cell responses as well as neutralizing antibodies. Altogether, these results have important meanings for the development of anti-HCV preventive vaccines and exemplify the flexibility and potential of our retrovirus-based platform in inducing broad cellular and humoral immune responses.

  12. Pediatric measles vaccine expressing a dengue antigen induces durable serotype-specific neutralizing antibodies to dengue virus.

    PubMed

    Brandler, Samantha; Lucas-Hourani, Marianne; Moris, Arnaud; Frenkiel, Marie-Pascale; Combredet, Chantal; Février, Michèle; Bedouelle, Hugues; Schwartz, Olivier; Desprès, Philippe; Tangy, Frédéric

    2007-12-12

    Dengue disease is an increasing global health problem that threatens one-third of the world's population. Despite decades of efforts, no licensed vaccine against dengue is available. With the aim to develop an affordable vaccine that could be used in young populations living in tropical areas, we evaluated a new strategy based on the expression of a minimal dengue antigen by a vector derived from pediatric live-attenuated Schwarz measles vaccine (MV). As a proof-of-concept, we inserted into the MV vector a sequence encoding a minimal combined dengue antigen composed of the envelope domain III (EDIII) fused to the ectodomain of the membrane protein (ectoM) from DV serotype-1. Immunization of mice susceptible to MV resulted in a long-term production of DV1 serotype-specific neutralizing antibodies. The presence of ectoM was critical to the immunogenicity of inserted EDIII. The adjuvant capacity of ectoM correlated with its ability to promote the maturation of dendritic cells and the secretion of proinflammatory and antiviral cytokines and chemokines involved in adaptive immunity. The protective efficacy of this vaccine should be studied in non-human primates. A combined measles-dengue vaccine might provide a one-shot approach to immunize children against both diseases where they co-exist.

  13. Neutral metoclopramide sensitizes cytotoxicity induced by ionizing radiation in SCID mice xenografted with a human brain astrocytoma.

    PubMed

    Hua, J; Olsson, A R; Pero, R W

    1997-12-10

    A formulation of metoclopramide (MCA) conformationally altered by neutralization of pH (nMCA, Neu-Sensamide) has been shown to have the same efficacy of enhancing the cytotoxicity of a single dose of 1 Gy radiation as acidic formulations (e.g., Primperan, Sensamide) in a human lung adenocarcinoma (H2981) xenografted into SCID mice. In the present study, 2 x 1 Gy radiation was combined with 2 x 2 mg nMCA/kg body weight injected 2 hr before radiation treatment for evaluation of radiosensitization in SCID mice xenografted with a human brain astrocytoma (T24). Given in this treatment schedule, nMCA alone at 2 mg/kg showed no cytotoxic effect on tumor growth in vivo. When combined with 2 x 1 Gy of radiation, however, the cytotoxicity was significantly increased as measured by tumor growth delay over the radiation-only-treated group. Furthermore, nMCA was absorbed into brains of mice and rats as efficiently as acidic MCA (aMCA) when analyzed 45 min after i.m. injection by high-performance liquid chromatography.

  14. ELPylated haemagglutinins produced in tobacco plants induce potentially neutralizing antibodies against H5N1 viruses in mice.

    PubMed

    Phan, Hoang T; Pohl, Julia; Floss, Doreen M; Rabenstein, Frank; Veits, Jutta; Le, Binh T; Chu, Ha H; Hause, Gerd; Mettenleiter, Thomas; Conrad, Udo

    2013-06-01

    Reducing the cost of vaccine production is a key priority for veterinary research, and the possibility of heterologously expressing antigen in plants provides a particularly attractive means of achieving this. Here, we report the expression of the avian influenza virus haemagglutinin (AIV HA) in tobacco, both as a monomer and as a trimer in its native and its ELPylated form. We firstly presented evidence to produce stabilized trimers of soluble HA in plants. ELPylation of these trimers does not influence the trimerization. Strong expression enhancement in planta caused by ELPylation was demonstrated for trimerized H5-ELP. ELPylated trimers could be purified by a membrane-based inverse transition cycling procedure with the potential of successful scale-up. The trimeric form of AIV HA was found to enhance the HA-specific immune response compared with the monomeric form. Plant-derived AIV HA trimers elicited potentially neutralizing antibodies interacting with both homologous virus-like particles from plants and heterologous inactivated AIV. ELPylation did not influence the functionality and the antigenicity of the stabilized H5 trimers. These data allow further developments including scale-up of production, purification and virus challenge experiments with the final goal to achieve suitable technologies for efficient avian flu vaccine production.

  15. Wound healing during hibernation by black bears (Ursus americanus) in the wild: elicitation of reduced scar formation.

    PubMed

    Iaizzo, Paul A; Laske, Timothy G; Harlow, Henry J; McClay, Carolyn B; Garshelis, David L

    2012-03-01

    Even mildly hypothermic body or limb temperatures can retard healing processes in mammals. Despite this, we observed that hibernating American black bears (Ursus americanus Pallas, 1780) elicit profound abilities in mounting inflammatory responses to infection and/or foreign bodies. In addition, they resolve injuries during hibernation while maintaining mildly hypothermic states (30-35 °C) and without eating, drinking, urinating or defecating. We describe experimental studies on free-ranging bears that document their abilities to completely resolve cutaneous cuts and punctures incurred during or prior to hibernation. We induced small, full-thickness cutaneous wounds (biopsies or incisions) during early denning, and re-biopsied sites 2-3 months later (near the end of denning). Routine histological methods were used to characterize these skin samples. All biopsied sites with respect to secondary intention (open circular biopsies) and primary intention (sutured sites) healed, with evidence of initial eschar (scab) formation, completeness of healed epidermis and dermal layers, dyskeratosis (inclusion cysts), and abilities to produce hair follicles. These healing abilities of hibernating black bears are a clear survival advantage to animals injured before or during denning. Bears are known to have elevated levels of hibernation induction trigger (delta-opioid receptor agonist) and ursodeoxycholic acid (major bile acid within plasma, mostly conjugated with taurine) during hibernation, which may relate to these wound-healing abilities. Further research as to the underlying mechanisms of wound healing during hibernation could have applications in human medicine. Unique approaches may be found to improve healing for malnourished, hypothermic, diabetic and elderly patients or to reduce scarring associated with burns and traumatic injuries.

  16. Determining causes of genetic isolation in a large carnivore (Ursus americanus) population to direct contemporary conservation measures

    PubMed Central

    Obbard, Martyn E.; Harnden, Matthew; McConnell, Sabine; Howe, Eric J.; Burrows, Frank G.; White, Bradley N.; Kyle, Christopher J.

    2017-01-01

    The processes leading to genetic isolation influence a population’s local extinction risk, and should thus be identified before conservation actions are implemented. Natural or human-induced circumstances can result in historical or contemporary barriers to gene flow and/or demographic bottlenecks. Distinguishing between these hypotheses can be achieved by comparing genetic diversity and differentiation in isolated vs. continuous neighboring populations. In Ontario, American black bears (Ursus americanus) are continuously distributed, genetically diverse, and exhibit an isolation-by-distance structuring pattern, except on the Bruce Peninsula (BP). To identify the processes that led to the genetic isolation of BP black bears, we modelled various levels of historical and contemporary migration and population size reductions using forward simulations. We compared simulation results with empirical genetic indices from Ontario black bear populations under different levels of geographic isolation, and conducted additional simulations to determine if translocations could help achieve genetic restoration. From a genetic standpoint, conservation concerns for BP black bears are warranted because our results show that: i) a recent demographic bottleneck associated with recently reduced migration best explains the low genetic diversity on the BP; and ii) under sustained isolation, BP black bears could lose between 70% and 80% of their rare alleles within 100 years. Although restoring migration corridors would be the most effective method to enhance long-term genetic diversity and prevent inbreeding, it is unrealistic to expect connectivity to be re-established. Current levels of genetic diversity could be maintained by successfully translocating 10 bears onto the peninsula every 5 years. Such regular translocations may be more practical than landscape restoration, because areas connecting the peninsula to nearby mainland black bear populations have been irreversibly modified

  17. Determining causes of genetic isolation in a large carnivore (Ursus americanus) population to direct contemporary conservation measures.

    PubMed

    Pelletier, Agnès; Obbard, Martyn E; Harnden, Matthew; McConnell, Sabine; Howe, Eric J; Burrows, Frank G; White, Bradley N; Kyle, Christopher J

    2017-01-01

    The processes leading to genetic isolation influence a population's local extinction risk, and should thus be identified before conservation actions are implemented. Natural or human-induced circumstances can result in historical or contemporary barriers to gene flow and/or demographic bottlenecks. Distinguishing between these hypotheses can be achieved by comparing genetic diversity and differentiation in isolated vs. continuous neighboring populations. In Ontario, American black bears (Ursus americanus) are continuously distributed, genetically diverse, and exhibit an isolation-by-distance structuring pattern, except on the Bruce Peninsula (BP). To identify the processes that led to the genetic isolation of BP black bears, we modelled various levels of historical and contemporary migration and population size reductions using forward simulations. We compared simulation results with empirical genetic indices from Ontario black bear populations under different levels of geographic isolation, and conducted additional simulations to determine if translocations could help achieve genetic restoration. From a genetic standpoint, conservation concerns for BP black bears are warranted because our results show that: i) a recent demographic bottleneck associated with recently reduced migration best explains the low genetic diversity on the BP; and ii) under sustained isolation, BP black bears could lose between 70% and 80% of their rare alleles within 100 years. Although restoring migration corridors would be the most effective method to enhance long-term genetic diversity and prevent inbreeding, it is unrealistic to expect connectivity to be re-established. Current levels of genetic diversity could be maintained by successfully translocating 10 bears onto the peninsula every 5 years. Such regular translocations may be more practical than landscape restoration, because areas connecting the peninsula to nearby mainland black bear populations have been irreversibly modified

  18. Neutral depletion versus repletion due to ionization

    SciTech Connect

    Fruchtman, A.; Makrinich, G.; Raimbault, J.-L.; Liard, L.; Rax, J.-M.; Chabert, P.

    2008-05-15

    Recent theoretical analyses which predicted unexpected effects of neutral depletion in both collisional and collisionless plasmas are reviewed. We focus on the depletion of collisionless neutrals induced by strong ionization of a collisionless plasma and contrast this depletion with the effect of strong ionization on thermalized neutrals. The collisionless plasma is analyzed employing a kinetic description. The collisionless neutrals and the plasma are coupled through volume ionization and wall recombination only. The profiles of density and pressure both of the plasma and of the neutral-gas and the profile of the ionization rate are calculated. It is shown that for collisionless neutrals the ionization results in neutral depletion, while when neutrals are thermalized the ionization induces a maximal neutral-density at the discharge center, which we call neutral repletion. The difference between the two cases stems from the relation between the neutral density and pressure. The pressure of the collisionless neutral-gas turns out to be maximal where its density is minimal, in contrast to the case of a thermalized neutral gas.

  19. Structure of monomeric Na-GST-3, a glutathione S-transferase from the major human hookworm parasite Necator americanus.

    PubMed

    Kelleher, Alan; Zhan, Bin; Asojo, Oluwatoyin A

    2013-08-01

    Necator americanus is the major cause of human hookworm infection, which is a global cause of anemia in the developing world. Ongoing efforts to control hookworm infection include the identification of candidate vaccine antigens as well as potential therapeutic targets from the infective L3 larval stages and adult stages of the parasite. One promising family of proteins are the adult-stage-secreted cytosolic glutathione S-transferases (GSTs). Nematode GSTs facilitate the inactivation and degradation of a variety of electrophilic substrates (drugs) via the nucleophilic addition of reduced glutathione. Parasite GSTs also play significant roles in multi-drug resistance and the modulation of host immune defense mechanisms. Here, the structure of Na-GST-3, one of three GSTs secreted by adult-stage N. americanus, is reported. Unlike most GST structures, the Na-GST-3 crystal contains a monomer in the asymmetric unit. However, the monomer forms a prototypical GST dimer across the crystallographic twofold. A glutathione from the fermentation process is bound to the monomer. The overall binding cavity of Na-GST-3 is reminiscent of that of other N. americanus GSTs and is larger and capable of binding a wider array of ligands than GSTs from organisms that have other major detoxifying mechanisms. Furthermore, despite having low sequence identity to the host GST, Na-GST-3 has a greater tertiary-structure similarity to human sigma-class GST than was observed for the other N. americanus GSTs.

  20. Trichinella Surveillance in Black Bears ( Ursus americanus ) from the Dehcho Region, Northwest Territories, Canada, 2002-15.

    PubMed

    Larter, Nicholas C; Elkin, Brett T; Forbes, Lorry B; Wagner, Brent; Allaire, Danny G

    2017-04-01

    We used muscle digestion to test black bears ( Ursus americanus ) from the southwestern Northwest Territories, Canada, for Trichinella. Results showed a prevalence of 4.1%. Some bears had infection intensities of more than one larva per gram of muscle tissue; this level in meat is considered to pose a human consumption safety risk.

  1. Biomarker analysis of American toad (Anaxyrus americanus) and grey tree frog (Hyla versicolor) tadpoles following exposure to atrazine.

    EPA Science Inventory

    The objective of the current study was to use a biomarker-based approach to investigate the influence of atrazine exposure on American toad (Anaxyrus americanus) and grey tree frog (Hyla versicolor) tadpoles. Atrazine is one of the most frequently detected herbicides in environme...

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

    USDA-ARS?s Scientific Manuscript database

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

  3. Biomarker analysis of American toad (Anaxyrus americanus) and grey tree frog (Hyla versicolor) tadpoles following exposure to atrazine.

    EPA Science Inventory

    The objective of the current study was to use a biomarker-based approach to investigate the influence of atrazine exposure on American toad (Anaxyrus americanus) and grey tree frog (Hyla versicolor) tadpoles. Atrazine is one of the most frequently detected herbicides in environme...

  4. The hidden history of the snowshoe hare, Lepus americanus: extensive mitochondrial DNA introgression inferred from multilocus genetic variation.

    PubMed

    Melo-Ferreira, José; Seixas, Fernando A; Cheng, Ellen; Mills, L Scott; Alves, Paulo C

    2014-09-01

    Hybridization drives the evolutionary trajectory of many species or local populations, and assessing the geographic extent and genetic impact of interspecific gene flow may provide invaluable clues to understand population divergence or the adaptive relevance of admixture. In North America, hares (Lepus spp.) are key species for ecosystem dynamics and their evolutionary history may have been affected by hybridization. Here we reconstructed the speciation history of the three most widespread hares in North America - the snowshoe hare (Lepus americanus), the white-tailed jackrabbit (L. townsendii) and the black-tailed jackrabbit (L. californicus) - by analysing sequence variation at eight nuclear markers and one mitochondrial DNA (mtDNA) locus (6240 bp; 94 specimens). A multilocus-multispecies coalescent-based phylogeny suggests that L. americanus diverged ~2.7 Ma and that L. californicus and L. townsendii split more recently (~1.2 Ma). Within L. americanus, a deep history of cryptic divergence (~2.0 Ma) was inferred, which coincides with major speciation events in other North American species. While the isolation-with-migration model suggested that nuclear gene flow was generally rare or absent among species or major genetic groups, coalescent simulations of mtDNA divergence revealed historical mtDNA introgression from L. californicus into the Pacific Northwest populations of L. americanus. This finding marks a history of past reticulation between these species, which may have affected other parts of the genome and influence the adaptive potential of hares during climate change.

  5. Neutral Particle Analyzer Vertically Scanning Measurements of MHD-induced Energetic Ion Redistribution or Loss in the National Spherical Torus Experiment

    SciTech Connect

    S.S. Medley, R. Andre, R.E. Bell, D.S. Darrow, C.W. Domier, E.D. Fredrickson, N.N. Gorelenkov, S.M. Kaye, B.P. LeBlanc, K.C. Lee, F.M. Levinton, D. Liu, N.C. Luhmann, Jr., J.E. Menard, H. Park, D. Stutman, A.L. Roquemore, K. Tritz, H. Yuh and the NSTX Team

    2007-11-15

    Observations of magneto-hydro-dynamic (MHD) induced redistribution or loss of energetic ions measured using the vertically scanning capability of the Neutral Particle Analyzer diagnostic on the National Spherical Torus Experiment (NSTX) are presented along with TRANSP and ORBIT code analysis of the results. Although redistribution or loss of energetic ions due to bursting fishbone-like and low-frequency (f ~ 10 kHz) kinktype MHD activity has been reported previously, the primary goal of this work is to study redistribution or loss due to continuous Alfvénic (f ~ 20 – 150 kHz) modes, a topic that heretofore has not been investigated in detail for NSTX plasmas. Initial indications are that the former drive energetic ion loss whereas the continuous Alfvénic modes only cause redistribution and the energetic ions remain confined.

  6. TRIGLYCERIDE-RICH LIPOPROTEIN LIPOLYSIS RELEASES NEUTRAL AND OXIDIZED FREE FATTY ACIDS THAT INDUCE ENDOTHELIAL CELL INFLAMMATION

    USDA-ARS?s Scientific Manuscript database

    Objective–Increased products of triglyceride-rich lipoprotein (TGRL) lipolysis provide a pro-inflammatory stimulus that may alter endothelial barrier function. To probe the mechanism of this lipolysis-induced dysfunction, we evaluated the pro-inflammatory potential of lipid classes derived from huma...

  7. A mimotope from a solid-phase peptide library induces a measles virus-neutralizing and protective antibody response.

    PubMed

    Steward, M W; Stanley, C M; Obeid, O E

    1995-12-01

    A solid-phase 8-mer random combinatorial peptide library was used to generate a panel of mimotopes of an epitope recognized by a monoclonal antibody to the F protein of measles virus (MV). An inhibition immunoassay was used to show that these peptides were bound by the monoclonal antibody with different affinities. BALB/c mice were coimmunized with the individual mimotopes and a T-helper epitope peptide (from MV fusion protein), and the reactivity of the induced anti-mimotope antibodies with the corresponding peptides and with MV was determined. The affinities of the antibodies with the homologous peptides ranged from 8.9 x 10(5) to 4.4 x 10(7) liters/mol. However, only one of the anti-mimotope antibodies cross-reacted with MV in an enzyme-linked immunosorbent assay and inhibited MV plaque formation. Coimmunization of mice with this mimotope and the T-helper epitope peptide induced an antibody response which conferred protection against fatal encephalitis induced following challenge with MV and with the structurally related canine distemper virus. These results indicate that peptide libraries can be used to identify mimotopes of conformational epitopes and that appropriate immunization with these mimotopes can induce protective antibody responses.

  8. Human cytomegalovirus hyperimmune globulin not only neutralizes HCMV infectivity, but also inhibits HCMV-induced intracellular NF-kappaB, Sp1, and PI3-K signaling pathways.

    PubMed

    Andreoni, Kenneth A; Wang, Xin; Huang, Shu-Mei; Huang, Eng-Shang

    2002-05-01

    Inhibition of virus-induced intracellular signaling pathways and viral infectivity are our ultimate goals in the development of effective antiviral agents to control human cytomegalovirus (HCMV) infections. The HCMV hyperimmune globulin may meet such criteria. In a human embryonic lung (HEL) fibroblast culture model, pretreatment of Towne strain HCMV with HCMV hyperimmune globulin was shown to inhibit viral infectivity successfully, as measured by a standard plaque assay. The extracellular viral titers and extracellular viral DNA, as measured by plaque assay and PCR, respectively, were also decreased. In addition, the HCMV hyperimmune globulin prevented HCMV from inducing the intracellular activation of NF-kappaB, Sp-1, and PI3-K signaling pathways. The PI3-K pathway was examined by following phosphorylation (activation) of two of its downstream kinases, Akt and p70S6K. HCMV hyperimmune globulin also prevented the production of immediate early, early, and late viral proteins. These studies show that HCMV hyperimmune globulin neutralization of HCMV prevents the earliest known events observed after viral envelope glycoproteins bind their cell membrane receptors, i.e., NF-kappaB, Sp-1 and PI3-K activation. This suggests that HCMV hyperimmune globulin not only can inhibit viral infectivity, but can also prevent the abnormal cellular signaling that may induce unwanted cellular proliferation or cytokine synthesis. Copyright 2002 Wiley-Liss, Inc.

  9. The neurosteroid pregnenolone sulfate neutralized the learning impairment induced by intrahippocampal nicotine in alcohol-drinking rats.

    PubMed

    Martín-García, E; Pallarès, M

    2005-01-01

    The effects of intrahippocampal administration of nicotine and the neurosteroids pregnenolone sulfate and allopregnanolone on acquiring the lever-press response and extinction in a Skinner box were examined using voluntary alcohol-drinking rats. A free-choice drinking procedure that implies early availability of the alcoholic solution (10% ethanol v/v+3% glucose w/v in distilled water) was used. Alcohol and control rats were deprived of food and assigned at random to six groups. Each group received two consecutive intrahippocampal (dorsal CA1) injections immediately after 1-h of drinking ethanol and before the free lever-press response shaping and extinction session. The groups were: saline-saline; saline-pregnenolone sulfate (5 ng, 24 microM); saline-allopregnanolone (0.2 microg, 1.26 microM); nicotine (4.6 microg, 20 mM)-saline; nicotine-pregnenolone sulfate; nicotine-allopregnanolone. Blood alcohol concentrations were assessed the day before conditioning. The combination of the oral self-administration of ethanol and the intrahippocampal injection of nicotine deteriorated the ability to acquire the lever-press response. This effect was neutralized by intrahippocampal pregnenolone sulfate (negative modulator of the GABA(A) receptor complex), and it was not affected by intrahippocampal allopregnanolone (positive GABA receptor complex A modulator). Pregnenolone sulfate and allopregnanolone had no effects per se on lever-press acquisition, neither in alcohol-drinking rats nor in controls. Alcohol consumption facilitated operant extinction just as anxiolytics that act as positive modulators of the GABA receptor complex A receptors do, possibly reducing the anxiety or aversion related to non-reinforcement. This effect was increased by intrahippocampal nicotine.

  10. Apoptosis induced by capsaicin in prostate PC-3 cells involves ceramide accumulation, neutral sphingomyelinase, and JNK activation.

    PubMed

    Sánchez, Ana Maria; Malagarie-Cazenave, Sophie; Olea, Nuria; Vara, Diana; Chiloeches, Antonio; Díaz-Laviada, Inés

    2007-11-01

    Numerous studies have recently focused on the anticarcinogenic, antimutagenic, or chemopreventive activities of the main pungent component of red pepper, capsaicin (N-vanillyl-8-methyl-1-nonenamide). We have previously shown that, in the androgen-independent prostate cancer PC-3 cells, capsaicin inhibits cell growth and induces apoptosis through reactive oxygen species (ROS) generation [Apoptosis 11 (2006) 89-99]. In the present study, we investigated the signaling pathways involved in the antiproliferative effect of capsaicin. Here, we report that capsaicin apoptotic effect was mediated by ceramide generation which occurred by sphingomyelin hydrolysis. Using siRNA, we demonstrated that N-SMase expression is required for the effect of capsaicin on prostate cell viability. We then investigated the role of MAP kinase cascades, extracellular signal-regulated protein kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 MAPK, in the antiproliferative effect of capsaicin, and we confirmed that capsaicin could activate ERK and JNK but not p38 MAPK. Pharmacological inhibition of JNK kinase, as well as inhibition of ROS by the reducing agent N-acetylcysteine, prevented ceramide accumulation and capsaicin-induced cell death. However, inhibition of ceramide accumulation by the SMase inhibitor D609 did not modify JNK activation. These data reveal JNK as an upstream regulator of ceramide production. Capsaicin-promoted activation of ERK was prevented with all the inhibitors tested. We conclude that capsaicin induces apoptosis in PC-3 cells via ROS generation, JNK activation, ceramide accumulation, and second, ERK activation.

  11. Signal enhancement of neutral He emission lines by fast electron bombardment of laser-induced He plasma

    SciTech Connect

    Suyanto, Hery; Pardede, Marincan; Hedwig, Rinda; Marpaung, Alion Mangasi; Ramli, Muliadi; Lie, Tjung Jie; Kurniawan, Koo Hendrik; Abdulmadjid, Syahrun Nur; Tjia, May On; Kagawa, Kiichiro

    2016-08-15

    A time-resolved spectroscopic study is performed on the enhancement signals of He gas plasma emission using nanosecond (ns) and picosecond (ps) lasers in an orthogonal configuration. The ns laser is used for the He gas plasma generation and the ps laser is employed for the ejection of fast electrons from a metal target, which serves to excite subsequently the He atoms in the plasma. The study is focused on the most dominant He I 587.6 nm and He I 667.8 nm emission lines suggested to be responsible for the He-assisted excitation (HAE) mechanism. The time-dependent intensity enhancements induced by the fast electrons generated with a series of delayed ps laser ablations are deduced from the intensity time profiles of both He emission lines. The results clearly lead to the conclusion that the metastable excited triplet He atoms are actually the species overwhelmingly produced during the recombination process in the ns laser-induced He gas plasma. These metastable He atoms are believed to serve as the major energy source for the delayed excitation of analyte atoms in ns laser-induced breakdown spectroscopy (LIBS) using He ambient gas.

  12. Signal enhancement of neutral He emission lines by fast electron bombardment of laser-induced He plasma

    NASA Astrophysics Data System (ADS)

    Suyanto, Hery; Pardede, Marincan; Hedwig, Rinda; Marpaung, Alion Mangasi; Ramli, Muliadi; Lie, Tjung Jie; Abdulmadjid, Syahrun Nur; Kurniawan, Koo Hendrik; Tjia, May On; Kagawa, Kiichiro

    2016-08-01

    A time-resolved spectroscopic study is performed on the enhancement signals of He gas plasma emission using nanosecond (ns) and picosecond (ps) lasers in an orthogonal configuration. The ns laser is used for the He gas plasma generation and the ps laser is employed for the ejection of fast electrons from a metal target, which serves to excite subsequently the He atoms in the plasma. The study is focused on the most dominant He I 587.6 nm and He I 667.8 nm emission lines suggested to be responsible for the He-assisted excitation (HAE) mechanism. The time-dependent intensity enhancements induced by the fast electrons generated with a series of delayed ps laser ablations are deduced from the intensity time profiles of both He emission lines. The results clearly lead to the conclusion that the metastable excited triplet He atoms are actually the species overwhelmingly produced during the recombination process in the ns laser-induced He gas plasma. These metastable He atoms are believed to serve as the major energy source for the delayed excitation of analyte atoms in ns laser-induced breakdown spectroscopy (LIBS) using He ambient gas.

  13. Novel synthetic plasmid and Doggybone™ DNA vaccines induce neutralizing antibodies and provide protection from lethal influenza challenge in mice

    PubMed Central

    Scott, Veronica L; Patel, Ami; Villarreal, Daniel O; Hensley, Scott E; Ragwan, Edwin; Yan, Jian; Sardesai, Niranjan Y; Rothwell, Paul J; Extance, Jonathan P; Caproni, Lisa J; Weiner, David B

    2015-01-01

    Nucleic acid-based vaccines (NAVs) are a promising alternative to conventional influenza vaccines with the potential to increase influenza vaccine availability due to their simplicity in design and rapid speed of production. NAVs can also target multiple influenza antigens and control flu variants. Traditionally NAVs have been DNA plasmids however, we are continuing to explore new methods that may enhance vaccine efficacy. Recently new focus has been on RNA cassettes as NAVs. RNA vaccines combine conceptual advantages in that they focus on delivery of only the coding cassette. However, RNA vaccines have a short half-life and cause interferon-induced fevers. Here we describe a new NAV approach where we study delivery of a linear DNA cassette [Doggybone™ linear closed DNA [(dbDNA™)] produced by an enzymatic process that yields an antigen expression cassette comprising a promoter, DNA antigen, poly A tail, and telomeric ends. This focused approach has many of the advantages of plasmid DNA as well as a minimal cassette size similar to RNA strategies. For this study, we characterized the specific CD4+ and CD8+ T cell responses and determined the hemagglutination inhibition (HI) titers induced by dbDNA™ and compared the responses with those of an optimized plasmid DNA (pDNA) vaccine encoding the same H1N1 influenza A/PR/8/34 HA gene. Immunizations with the constructs resulted in similar humoral and cellular immune responses. Both constructs induced high-titer HI antibodies and fully protected animals from lethal viral challenge. The data obtained from this study provides important validation for further development of novel vector approaches. PMID:26091432

  14. A goat poxvirus-vectored peste-des-petits-ruminants vaccine induces long-lasting neutralization antibody to high levels in goats and sheep.

    PubMed

    Chen, Weiye; Hu, Sen; Qu, Linmao; Hu, Qianqian; Zhang, Qian; Zhi, Haibing; Huang, Kehe; Bu, Zhigao

    2010-07-05

    Recombinant capripoxvirus (CPV) is a promising candidate differentiating infected from vaccinated animals (DIVA) vaccine against peste-des-petits-ruminants (PPR). In order for recombinant CPV to be successfully used in the field, there should exist dependable indicators for quality control of vaccine products, surveillance and vaccination evaluation. Viral neutralization antibody (VNA) is correlated to protection against PPR and is a technically feasible indicator for this purpose. The immunogenicity of this vectored vaccine in goats and sheep, however, has not been fully evaluated. In this study, we generated two recombinant CPV viruses, rCPV-PPRVH and rCPV-PPRVF, that express PPR virus (PPRV) glycoproteins H and F, respectively. Vaccination studies with different dosages of recombinant viruses showed that rCPV-PPRVH was a more potent inducer of PPRV VNA than rCPV-PPRVF. One dose of rCPV-PPRVH was enough to seroconvert 80% of immunized sheep. A second dose induced significantly higher PPRV VNA titers. There was no significant difference in PPRV VNA responses between goats and sheep. Subcutaneous inoculation also induced a significant PPRV VNA response. PPRV VNA could be detected for over 6 months in more than 80% of vaccinated goats and sheep. Boost vaccination at 6-month intervals induced significant re-boost efficacy of PPRV VNA in goats and sheep. More over, two doses of rCPV-PPRVH could completely overcome the interference caused by pre-existing immunity to the CPV vaccine backbone in animals. Vaccination with rCPV-PPRVH also protected goats from virulent CPV challenge. Our results demonstrate that VNA can serve as a dependent indicator for effective vaccination and immune protection of animals in the field. The recombinant CPV vaccine used in our studies could be a practical and useful candidate DIVA vaccine in countries where PPR newly emerges or where stamp-out plans are yet to be implemented. Copyright 2010 Elsevier Ltd. All rights reserved.

  15. Systemic Neutralization of Interleukin-8 Markedly Reduces Neutrophilic Pleocytosis during Experimental Lipopolysaccharide-Induced Meningitis in Rabbits

    PubMed Central

    Dumont, Rachel A.; Car, Bruce D.; Voitenok, Nikolai N.; Junker, Ursula; Moser, Bernhard; Zak, Oto; O'Reilly, Terence

    2000-01-01

    Interleukin-8 (IL-8) is elevated in the cerebrospinal fluid (CSF) of patients with meningitis and is proposed to participate in subarachnoid-space pleocytosis. However, intracisternal injection of IL-8 into rabbits failed to induce indices typical of meningitis (leukocyte, tumor necrosis factor, or protein accumulation in the CSF or histopathological changes), indicating that merely increasing the CSF level of this chemokine is insufficient to induce inflammation in this anatomical site. IL-8 treatment did not affect inflammatory responses to subsequently intracisternally administered lipopolysaccharide (LPS). IL-8 was chemotactic for rabbit neutrophils in vitro, and subcutaneous injection of IL-8 (diluted in buffer or CSF) proved the in vivo activity of this peptide and suggested the absence of an IL-8 inhibitor in normal rabbit CSF. LPS-dependent pleocytosis was only slightly diminished by intracisternally administered murine anti-rabbit IL-8 monoclonal antibody (MAb) WS-4 but was dramatically reduced by intravenously administered MAb. Therefore, elevated CSF IL-8 levels may contribute to, but cannot solely account for, neutrophil influx into the subarachnoid space during meningitis. However, inhibition of IL-8 activity of the bloodstream side of the blood-brain barrier effectively reduces pleocytosis, indicating a central role of IL-8 in neutrophil influx into CSF during bacterial meningitis. Thus, inhibition of IL-8 is a possible therapeutic target for adjunct treatment of meningitis. PMID:10992482

  16. Heat-induced denaturation and aggregation of ovalbumin at neutral pH described by irreversible first-order kinetics.

    PubMed

    Weijers, Mireille; Barneveld, Peter A; Cohen Stuart, Martien A; Visschers, Ronald W

    2003-12-01

    The heat-induced denaturation kinetics of two different sources of ovalbumin at pH 7 was studied by chromatography and differential scanning calorimetry. The kinetics was found to be independent of protein concentration and salt concentration, but was strongly dependent on temperature. For highly pure ovalbumin, the decrease in nondenatured native protein showed first-order dependence. The activation energy obtained with different techniques varied between 430 and 490 kJ*mole(-1). First-order behavior was studied in detail using differential scanning calorimetry. The calorimetric traces were irreversible and highly scan rate-dependent. The shape of the thermograms as well as the scan rate dependence can be explained by assuming that the thermal denaturation takes place according to a simplified kinetic process where N is the native state, D is denatured (or another final state) and k a first-order kinetic constant that changes with temperature, according to the Arrhenius equation. A kinetic model for the temperature-induced denaturation and aggregation of ovalbumin is presented. Commercially obtained ovalbumin was found to contain an intermediate-stable fraction (IS) of about 20% that was unable to form aggregates. The denaturation of this fraction did not satisfy first-order kinetics.

  17. Neutralization of MMP-2 and TNFR1 Regulates the Severity of S. aureus-Induced Septic Arthritis by Differential Alteration of Local and Systemic Proinflammatory Cytokines in Mice.

    PubMed

    Sultana, Sahin; Adhikary, Rana; Bishayi, Biswadev

    2017-03-22

    Despite advancement in the field of antibiotics septic arthritis remains a serious concern till date. Staphylococcus aureus is the most common bacterium that causes septic arthritis. Severity of this disease is directly correlated with chronic inflammation induced by proinflammatory cytokines like TNF-α, interleukin (IL)-1β, IL-6, and induction of matrix metalloproteinases (MMPs) including MMP-2. The objective of our study was to evaluate the role of MMP-2 and tumor necrosis factor receptor 1 (TNFR1) in the pathogenesis of S. aureus infection-induced septic arthritis. Mice were infected with live S. aureus (5 × 10(6) cells/ml) followed by administration of MMP-2 inhibitor and TNFR1 antibody. Arthritis index showed highest reduction in severity of arthritis in mice treated with both MMP-2 inhibitor and TNFR1 antibody after infection. Combined neutralization of MMP-2 and TNFR1 led to marked diminution in bacterial count in the combined group. Lowest levels of pro inflammatory cytokines like TNF-α, IL-1β, IL-6, and IFN-γ were observed in both serum and synovial tissues indicating maximum protection in S. aureus arthritis during combination treatment. Increment in the level of IL-10 in the combination group could be positively correlated with the recovery of arthritis. Similarly, expressions of COX-2 and iNOS, markers of acute inflammation were also significantly reduced in the combination group due to resolution of inflammation. Levels of O2(.-) and NO also showed a significant fall in case of the group treated with MMP-2 inhibitor and TNFR1 antibody both. Neutralization of both MMP-2 and TNFR1 caused rapid decline in recruitment of neutrophil and macrophages in the synovial tissues as evident from reduced MPO and MCP-1 levels, respectively, compared to other groups. Overall, it can be suggested that administration of MMP-2 inhibitor and TNFR1 antibody in combination is protective against the severity of inflammation and cartilage destruction associated with S

  18. A novel high mobility group box 1 neutralizing chimeric antibody attenuates drug‐induced liver injury and postinjury inflammation in mice

    PubMed Central

    Lea, Jonathan D.; Sowinska, Agnieszka; Ottosson, Lars; Fürst, Camilla Melin; Steen, Johanna; Aulin, Cecilia; Clarke, Joanna I.; Kipar, Anja; Klevenvall, Lena; Yang, Huan; Palmblad, Karin; Park, B. Kevin; Tracey, Kevin J.; Blom, Anna M.; Andersson, Ulf

    2016-01-01

    Acetaminophen (APAP) overdoses are of major clinical concern. Growing evidence underlines a pathogenic contribution of sterile postinjury inflammation in APAP‐induced acute liver injury (APAP‐ALI) and justifies development of anti‐inflammatory therapies with therapeutic efficacy beyond the therapeutic window of the only current treatment option, N‐acetylcysteine (NAC). The inflammatory mediator, high mobility group box 1 (HMGB1), is a key regulator of a range of liver injury conditions and is elevated in clinical and preclinical APAP‐ALI. The anti‐HMGB1 antibody (m2G7) is therapeutically beneficial in multiple inflammatory conditions, and anti‐HMGB1 polyclonal antibody treatment improves survival in a model of APAP‐ALI. Herein, we developed and investigated the therapeutic efficacy of a partly humanized anti‐HMGB1 monoclonal antibody (mAb; h2G7) and identified its mechanism of action in preclinical APAP‐ALI. The mouse anti‐HMGB1 mAb (m2G7) was partly humanized (h2G7) by merging variable domains of m2G7 with human antibody‐Fc backbones. Effector function‐deficient variants of h2G7 were assessed in comparison with h2G7 in vitro and in preclinical APAP‐ALI. h2G7 retained identical antigen specificity and comparable affinity as m2G7. 2G7 treatments significantly attenuated APAP‐induced serum elevations of alanine aminotransferase and microRNA‐122 and completely abrogated markers of APAP‐induced inflammation (tumor necrosis factor, monocyte chemoattractant protein 1, and chemokine [C‐X‐C motif] ligand 1) with prolonged therapeutic efficacy as compared to NAC. Removal of complement and/or Fc receptor binding did not affect h2G7 efficacy. Conclusion: This is the first report describing the generation of a partly humanized HMGB1‐neutralizing antibody with validated therapeutic efficacy and with a prolonged therapeutic window, as compared to NAC, in APAP‐ALI. The therapeutic effect was mediated by HMGB1 neutralization and

  19. Production and characterization of recombinant vitellogenesis-inhibiting hormone from the American lobster Homarus americanus.

    PubMed

    Ohira, Tsuyoshi; Okumura, Takuji; Suzuki, Michio; Yajima, Yosuke; Tsutsui, Naoaki; Wilder, Marcy N; Nagasawa, Hiromichi

    2006-06-01

    Recombinant peptides related to vitellogenesis-inhibiting hormone (VIH) of the American lobster Homarus americanus were expressed in bacterial cells, and then purified after being allowed to refold. Biological activities of the recombinant VIHs having an amidated C-terminus (rHoa-VIH-amide) and a free carboxyl-terminus (rHoa-VIH-OH) were examined using an ovarian fragment incubation system derived from the kuruma prawn, Marsupenaeus japonicus. The rHoa-VIH-amide significantly reduced vitellogenin mRNA levels in the ovary, while rHoa-VIH-OH had no effect. This is the first report that describes the production of a crustacean VIH having biological activity and the importance of the C-terminal amidation for its vitellogenesis-inhibiting activity.

  20. Laryngeal squamous cell carcinoma in a North American black bear (Ursus americanus).

    PubMed

    Rotstein, David S; Govett, Pamela; Wolfe, Barbara

    2005-09-01

    A 10-yr-old female black bear (Ursus americanus) at the North Carolina Zoological Park presented with a 2-wk history of intermittent coughing and stertor. The animal was anesthetized for examination, and at intubation, a large mass associated with the tonsils and larynx was observed obstructing the airway. Cardiopulmonary collapse occurred during the procedure, and efforts at resuscitation were ineffective. At necropsy, an 8 cm times 6 cm times 5 cm mass was present in the larynx, infiltrating the epiglottis and obstructing the airway. Regional lymph nodes were mildly enlarged and contained tan inspissated substance. Histologically, the neoplasm was composed of nests of keratinizing squamous epithelial cells with evidence of lymphatic invasion and metastases to regional lymph nodes.

  1. Naturally occurring bacteraemia in American lobsters, Homarus americanus Milne-Edwards, in Long Island Sound.

    PubMed

    Bartlett, S L; Wooster, G A; Sokolowski, M S; Dove, A D M; Bowser, P R

    2008-01-01

    The health status of the American lobster, Homarus americanus Milne-Edwards, in Long Island Sound (LIS) has been in decline, with seasonal mortality events occurring since 1998. In order to assess the potential effects of environmental conditions on lobster health via haemolymph analysis, lobsters collected from various sites in LIS were examined and sampled while concurrent environmental data (water temperature, salinity and dissolved oxygen) were recorded. The pH of the haemolymph of each lobster was tested, followed by a collection of haemolymph for serum biochemistry analysis and bacterial culture. This report focuses on the results of the bacterial sampling. The majority of bacteria cultured were opportunistic pathogens commonly found in the environment, including some that are associated with sewage and pollution. The prevalence of bacteraemia was correlated with the site of collection, the month in which the lobsters were sampled, and water temperature.

  2. Sublethal abnormalities in embryos of winter flounder, Psuedopleuronectes americanus, from Long Island Sound

    SciTech Connect

    Perry, D.M.; Hughes, J.B.; Herbert, A.T. )

    1991-09-01

    Sublethal abnormalities were examined in developing embryos of the winter flounder, Pseudopleuronectes americanus, as part of a study of its early reproductive success in Long Island Sound and two sites in Boston Harbor. These sites represented varying levels of anthropogenic contamination that were possibly affecting reproduction, as ascertained by chromosomal abnormalities and lowered developmental rates in the embryos. Effort was focused on examination of blastula and tail-bud embryos from field-captured females spawned at the laboratory. Abnormalities observed included evidence of cytotoxicity and chromosome damage. Embryos of fish New Haven were usually the most aberrant, while embryos from other sites, notably Hempstead, Shoreham, and both Boston Harbor stations, showed subtle indications of abnormality.

  3. 'Candidatus Liberibacter americanus', associated with citrus huanglongbing (greening disease) in São Paulo State, Brazil.

    PubMed

    Teixeira, Diva do Carmo; Saillard, Colette; Eveillard, Sandrine; Danet, Jean Luc; da Costa, Paulo Inácio; Ayres, Antonio Juliano; Bové, Joseph

    2005-09-01

    , were found to have identical or almost identical sequences. In contrast, the intergenic regions of the SPS-HLB liberibacter, 'Ca. L. asiaticus' and 'Ca. L. africanus' had quite different sequences, with similarity between 66.0 and 79.5%. These results confirm that the SPS-HLB liberibacter is a novel species for which the name 'Candidatus Liberibacter americanus' is proposed. Like the African and the Asian liberibacters, the 'American' liberibacter is restricted to the sieve tubes of the citrus host. The liberibacter could also be detected by PCR amplification of the 16S rRNA gene in Diaphorina citri, the psyllid vector of 'Ca. L. asiaticus', suggesting that this psyllid is also a vector of 'Ca. L. americanus' in SPS. 'Ca. L. americanus' was detected in 216 of 218 symptomatic leaf samples from 47 farms in 35 municipalities, while 'Ca. L. asiaticus' was detected in only 4 of the 218 samples, indicating that 'Ca. L. americanus' is the major cause of HLB in SPS.

  4. X-ray-induced DNA double-strand breaks in mouse l1210 cells: a new computational method for analyzing neutral filter elution data.

    PubMed

    Cedervall, Björn; Edgren, Margareta R; Lewensohn, Rolf

    2003-04-01

    The aim of this article is to present a method for studying the shape of the dose and repair responses for X-ray-induced double-strand breaks (DSBs) as measured by neutral filter elution (NFE). The approach is closely related to a method we developed for the use of specific molecular size markers and used for determination of the absolute number of randomly distributed radiation-induced DSBs by pulsed-field gel electrophoresis (PFGE). Mouse leukemia L1210 cells were X-irradiated with 0-50 Gy. Samples were then evaluated both with PFGE and with NFE. Assuming that with both migration (PFGE) and elution (NFE), a heterogeneous population of double-stranded DNA fragments will start with the smallest fragments and proceed with increasingly larger fragments, it is possible to match the migration behavior of fractions of fragments smaller than a certain size to the fraction eluted at a specific time. This assumption does not exclude the possibility of DNA being sheared in the NFE filter. The yield, as determined by the size markers in PFGE, was used to find the corresponding elution times in the NFE experiment. These experimentally used elution times could then reversely be interpreted as size markers which finally were used to calculate DSBs/Mbp as a function of X-ray dose. The resulting lines were almost straight. The data were also plotted as relative elution and showed that, as expected, the dose response then appears with a more pronounced sigmoid shape.

  5. The Novel Cannabinoid CB1 Receptor Neutral Antagonist AM4113 Suppresses Food Intake and Food-Reinforced Behavior but Does not Induce Signs of Nausea in Rats

    PubMed Central

    Sink, Kelly S; McLaughlin, Peter J; Wood, Jodi Anne T; Brown, Cara; Fan, Pusheng; Vemuri, V Kiran; Pang, Yan; Olzewska, Teresa; Thakur, Ganesh A; Makriyannis, Alex; Parker, Linda A; Salamone, John D

    2013-01-01

    Drugs that interfere with cannabinoid CB1 transmission suppress various food-motivated behaviors, and it has been suggested that such drugs could be useful as appetite suppressants. Biochemical studies indicate that most of these drugs assessed thus far have been CB1 inverse agonists, and although they have been shown to suppress food intake, they also appear to induce nausea and malaise. The present studies were undertaken to characterize the behavioral effects of AM4113, which is a CB1 neutral antagonist, and to examine whether this drug can reduce food-reinforced behaviors and feeding on diets with varying macronutrient compositions. Biochemical data demonstrated that AM4113 binds to CB1 receptors, but does not show inverse agonist properties (ie no effects on cyclic-AMP production). In tests of spontaneous locomotion and analgesia, AM4113 reversed the effects of the CB1 agonist AM411. AM4113 suppressed food-reinforced operant responding with rats responding on fixed ratio (FR) 1 and 5 schedules of reinforcement in a dose-dependent manner, and also suppressed feeding on high-fat, high-carbohydrate, and lab chow diets. However, in the same dose range that suppressed feeding, AM4113 did not induce conditioned gaping, which is a sign of nausea and food-related malaise in rats. These results suggest that AM4113 may decrease appetite by blocking endogenous cannabinoid tone, and that this drug may be less associated with nausea than CB1 inverse agonists. PMID:17581535

  6. Identification and cardiotropic actions of sulfakinin peptides in the American lobster Homarus americanus.

    PubMed

    Dickinson, Patsy S; Stevens, Jake S; Rus, Szymon; Brennan, Henry R; Goiney, Christopher C; Smith, Christine M; Li, Lingjun; Towle, David W; Christie, Andrew E

    2007-07-01

    In arthropods, a group of peptides possessing a -Y((SO3H))GHM/LRFamide carboxy-terminal motif have been collectively termed the sulfakinins. Sulfakinin isoforms have been identified from numerous insect species. In contrast, members of this peptide family have thus far been isolated from just two crustaceans, the penaeid shrimp Penaeus monodon and Litopenaeus vannamei. Here, we report the identification of a cDNA encoding prepro-sulfakinin from the American lobster Homarus americanus. Two sulfakinin-like sequences were identified within the open-reading frame of the cDNA. Based on modifications predicted by peptide modeling programs, and on homology to the known isoforms of sulfakinin, particularly those from shrimp, the mature H. americanus sulfakinins were hypothesized to be pEFDEY((SO3H))GHMRFamide (Hoa-SK I) and GGGEY((SO3H))DDY((SO3H))GHLRFamide (Hoa-SK II). Hoa-SK I is identical to one of the previously identified shrimp sulfakinins, while Hoa-SK II is a novel isoform. Exogenous application of either synthetic Hoa-SK I or Hoa-SK II to the isolated lobster heart increased both the frequency and amplitude of spontaneous heart contractions. In preparations in which spontaneous contractions were irregular, both peptides increased the regularity of the heartbeat. Our study provides the first molecular characterization of a sulfakinin-encoding cDNA from a crustacean, as well as the first demonstration of bioactivity for native sulfakinins in this group of arthropods.

  7. Structure of a two-CAP-domain protein from the human hookworm parasite Necator americanus

    SciTech Connect

    Asojo, Oluwatoyin A.

    2011-05-01

    The first structure of a two-CAP-domain protein, Na-ASP-1, from the major human hookworm parasite N. americanus refined to a resolution limit of 2.2 Å is presented. Major proteins secreted by the infective larval stage hookworms upon host entry include Ancylostoma secreted proteins (ASPs), which are characterized by one or two CAP (cysteine-rich secretory protein/antigen 5/pathogenesis related-1) domains. The CAP domain has been reported in diverse phylogenetically unrelated proteins, but has no confirmed function. The first structure of a two-CAP-domain protein, Na-ASP-1, from the major human hookworm parasite Necator americanus was refined to a resolution limit of 2.2 Å. The structure was solved by molecular replacement (MR) using Na-ASP-2, a one-CAP-domain ASP, as the search model. The correct MR solution could only be obtained by truncating the polyalanine model of Na-ASP-2 and removing several loops. The structure reveals two CAP domains linked by an extended loop. Overall, the carboxyl-terminal CAP domain is more similar to Na-ASP-2 than to the amino-terminal CAP domain. A large central cavity extends from the amino-terminal CAP domain to the carboxyl-terminal CAP domain, encompassing the putative CAP-binding cavity. The putative CAP-binding cavity is a characteristic cavity in the carboxyl-terminal CAP domain that contains a His and Glu pair. These residues are conserved in all single-CAP-domain proteins, but are absent in the amino-terminal CAP domain. The conserved His residues are oriented such that they appear to be capable of directly coordinating a zinc ion as observed for CAP proteins from reptile venoms. This first structure of a two-CAP-domain ASP can serve as a template for homology modeling of other two-CAP-domain proteins.

  8. Role of neutralizing anti-murine interleukin-17A monoclonal antibody on chronic ozone-induced airway inflammation in mice.

    PubMed

    Zhang, Min; Fei, Xia; Zhang, Guo-Qing; Zhang, Peng-Yu; Li, Feng; Bao, Wu-Ping; Zhang, Ying-Ying; Zhou, Xin

    2016-10-01

    Exposure to ozone has led to airway inflammation and airway hyperresponsiveness, which potential mechanisms relate to ozone-induced oxidative stress. IL-17 is a growing target for autoimmune and inflammatory diseases. The aim of the study was to examine the inhibitory effects of anti-murine interleukin-17A monoclonal antibody (IL-17mAb) on adverse effects of ozone which are noted above. After C57/BL6 mice were exposed to ozone (2.5ppm; 3h) for 12 times over 6 weeks, IL-17mAb, PBS was intraperitoneally injected into mice 1h after ozone or air exposure for 6 weeks and mice were studied 24h after final exposure, monitoring bronchial responsiveness, airway inflammatory cells, lung histology, levels of neutrophil-related chemokine and proinflammatory cytokines in bronchoalveolar lavage (BAL) fluid and serum, the expression of IL-17A mRNA and protein, glucocorticoid receptors (GR), and the phosphorylation of p38MAPK in lung tissues. The administration of IL-17mAb reduced the ozone-induced increases in total cells, especially neutrophils; decreased levels of cytokines, including IL-8 in BAL fluid, IL-8 and IL-17A in serum; mitigated the severity of airway hyperresponsiveness; attenuated lung inflammation scores and histologic analysis confirmed the suppression of lung inflammation, compared with the administration of a control PBS. Exposure to ozone results in increases in IL-17A production rate, mRNA and protein levels of IL-17A and the protein level of GR. These effects were halted and reversed by IL-17mAb treatment. Furthermore, IL-17mAb also reduced the phosphorylation of p38MAPK. Therefore, we conclude that IL-17mAb may be a useful therapy in ozone-related diseases, including COPD.

  9. Heat-induced denaturation and aggregation of ovalbumin at neutral pH described by irreversible first-order kinetics

    PubMed Central

    Weijers, Mireille; Barneveld, Peter A.; Cohen Stuart, Martien A.; Visschers, Ronald W.

    2003-01-01

    The heat-induced denaturation kinetics of two different sources of ovalbumin at pH 7 was studied by chromatography and differential scanning calorimetry. The kinetics was found to be independent of protein concentration and salt concentration, but was strongly dependent on temperature. For highly pure ovalbumin, the decrease in nondenatured native protein showed first-order dependence. The activation energy obtained with different techniques varied between 430 and 490 kJ•mole−1. First-order behavior was studied in detail using differential scanning calorimetry. The calorimetric traces were irreversible and highly scan rate-dependent. The shape of the thermograms as well as the scan rate dependence can be explained by assuming that the thermal denaturation takes place according to a simplified kinetic process \\documentclass[10pt]{article} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{pmc} \\usepackage[Euler]{upgreek} \\pagestyle{empty} \\oddsidemargin -1.0in \\begin{document} \\begin{equation*}N\\;{ \\,\\substack{ ^{{\\mathit{k}}} \\\\ {\\rightarrow} \\\\ }\\, }\\;D\\end{equation*}\\end{document} where N is the native state, D is denatured (or another final state) and k a first-order kinetic constant that changes with temperature, according to the Arrhenius equation. A kinetic model for the temperature-induced denaturation and aggregation of ovalbumin is presented. Commercially obtained ovalbumin was found to contain an intermediate-stable fraction (IS) of about 20% that was unable to form aggregates. The denaturation of this fraction did not satisfy first-order kinetics. PMID:14627731

  10. Laser Induced Emission Spectroscopy of Cold and Isolated Neutral PAHs and PANH: Implications for the red rectangle emission

    NASA Astrophysics Data System (ADS)

    Bejaoui, Salma; Salama, Farid; Sciamma O'Brien, Ella

    2016-06-01

    Blue luminescence (BL) in the emission spectra of the red rectangle centered on the bright star HD44179 is recently reported by Vijh et al [1]. This results is consistent with the broad band polarization measurements obtained in 1980 by Schmidt et al. Both experimental and theoretical studies support that BL emission could be attributed the luminescence of Polycyclic Aromatic Hydrocarbon (PAH) excited with ultraviolet light from the center of the star [4 and reference therein]. The abundance on N to C in the interstellar medium suggest also that nitrogen substituted PAH (PANH) are likely abundant in the interstellar medium [3]. They exhibit similar features as PAHs and could contribute to the unidentified spectral bands. Comparing the BL to laboratory spectra obtained on similar environment is crucial for the identification of interstellar molecules. We present in this works the absorption and the laser induced emission spectra of several isolated and cold PAHs and PANHs. Laser induced emission was performed first to PAHs and PANHs isolated in Argon matrix at 10 K. Then, measurements are performed with the supersonic jet technique of the COSmIC laboratory facility at NASA Ames. We focus, here, on the emission spectra (fluorescence and (or) phosphorescence) of these molecules and we discuss their contributions to the blue luminescence emission in the Red Rectangle nebula.[1] Vijh,U.P., Witt. A.N. & Gordon,K.D, APJ, 606, L69 (2004)[2] Schmidt, G. D., Cohen, M. & Margon, B., ApJ, 239L.133S (1980)[3] Spitzer, L., Physical Processes in the Interstellar Medium (New York Wiley-Interscience) (1978)[4] Salama, F., Galazutdinov, G. A., Kre lowski, J., Allamandola, L. J., & Musaev, F. A. ApJ, 526,(1999)

  11. Specific sequences commonly found in the V3 domain of HIV-1 subtype C isolates affect the overall conformation of native Env and induce a neutralization-resistant phenotype independent of V1/V2 masking.

    PubMed

    Salomon, Aidy; Krachmarov, Chavdar; Lai, Zhong; Honnen, William; Zingman, Barry S; Sarlo, Julie; Gorny, Miroslaw K; Zolla-Pazner, Susan; Robinson, James E; Pinter, Abraham

    2014-01-05

    Primary HIV-1 isolates are relatively resistant to neutralization by antibodies commonly induced after infection or vaccination. This is generally attributed to masking of sensitive epitopes by the V1/V2 domain and/or glycans situated at various positions in Env. Here we identified a novel masking effect mediated by subtype C-specific V3 sequences that contributes to the V1/V2-independent and glycan-independent neutralization resistance of chimeric and primary Envs to antibodies directed against multiple neutralization domains. Positions at several conserved charged and hydrophobic sites in the V3 crown and stem were also shown to affect neutralization phenotype. These results indicated that substitutions typically present in subtype C and related V3 sequences influence the overall conformation of native Env in a way that occludes multiple neutralization targets located both within and outside of the V3 domain, and may reflect an alternative mechanism for neutralization resistance that is particularly active in subtype C and related isolates.

  12. A Vaccine of L2 Epitope Repeats Fused with a Modified IgG1 Fc Induced Cross-Neutralizing Antibodies and Protective Immunity against Divergent Human Papillomavirus Types

    PubMed Central

    Zhang, Ting; Liu, Yanchun; Xie, Xixiu; Wang, Zhirong; Xu, Xuemei

    2014-01-01

    Current human papillomavirus (HPV) major capsid protein L1 virus-like particles (VLPs)-based vaccines in clinic induce strong HPV type-specific neutralizing antibody responses. To develop pan-HPV vaccines, here, we show that the fusion protein E3R4 consisting of three repeats of HPV16 L2 aa 17–36 epitope (E3) and a modified human IgG1 Fc scaffold (R4) induces cross-neutralizing antibodies and protective immunity against divergent HPV types. E3R4 was expressed as a secreted protein in baculovirus expression system and could be simply purified by one step Protein A affinity chromatography with the purity above 90%. Vaccination of E3R4 formulated with Freunds adjuvant not only induced cross-neutralizing antibodies against HPV pseudovirus types 16, 18, 45, 52, 58, 6, 11 and 5 in mice, but also protected mice against vaginal challenges with HPV pseudovirus types 16, 45, 52, 58, 11 and 5 for at least eleven months after the first immunization. Moreover, vaccination of E3R4 formulated with FDA approved adjuvant alum plus monophosphoryl lipid A also induced cross-neutralizing antibodies against HPV types 16, 18 and 6 in rabbits. Thus, our results demonstrate that delivery of L2 antigen as a modified Fc-fusion protein may facilitate pan-HPV vaccine development. PMID:24802101

  13. Toxin-neutralizing antibodies protect against Clostridium perfringens-induced necrosis in an intestinal loop model for bovine necrohemorrhagic enteritis.

    PubMed

    Goossens, Evy; Verherstraeten, Stefanie; Valgaeren, Bonnie R; Pardon, Bart; Timbermont, Leen; Schauvliege, Stijn; Rodrigo-Mocholí, Diego; Haesebrouck, Freddy; Ducatelle, Richard; Deprez, Piet R; Van Immerseel, Filip

    2016-06-13

    Bovine necrohemorrhagic enteritis is caused by Clostridium perfringens type A. Due to the rapid progress and fatal outcome of the disease, vaccination would be of high value. In this study, C. perfringens toxins, either as native toxins or after formaldehyde inactivation, were evaluated as possible vaccine antigens. We determined whether antisera raised in calves against these toxins were able to protect against C. perfringens challenge in an intestinal loop model for bovine necrohemorrhagic enteritis. Alpha toxin and perfringolysin O were identified as the most immunogenic proteins in the vaccine preparations. All vaccines evoked a high antibody response against the causative toxins, alpha toxin and perfringolysin O, as detected by ELISA. All antibodies were able to inhibit the activity of alpha toxin and perfringolysin O in vitro. However, the antibodies raised against the native toxins were more inhibitory to the C. perfringens-induced cytotoxicity (as tested on bovine endothelial cells) and only these antibodies protected against C. perfringens challenge in the intestinal loop model. Although immunization of calves with both native and formaldehyde inactivated toxins resulted in high antibody titers against alpha toxin and perfringolysin O, only antibodies raised against native toxins protect against C. perfringens challenge in an intestinal loop model for bovine necrohemorrhagic enteritis.

  14. Neutralizing Interleukin-1β (IL-1β) Induces β-Cell Survival by Maintaining PDX1 Protein Nuclear Localization*

    PubMed Central

    Ardestani, Amin; Sauter, Nadine S.; Paroni, Federico; Dharmadhikari, Gitanjali; Cho, Jae-Hyoung; Lupi, Roberto; Marchetti, Piero; Oberholzer, José; Conte, Julie Kerr; Maedler, Kathrin

    2011-01-01

    The transcription factor PDX1 plays a critical role during β-cell development and in glucose-induced insulin gene transcription in adult β-cells. Acute glucose exposure leads to translocalization of PDX1 to the nucleoplasm, whereas under conditions of oxidative stress, PDX1 shuttles from the nucleus to the cytosol. Here we show that cytosolic PDX1 expression correlated with β-cell failure in diabetes. In isolated islets from patients with type 2 diabetes and from diabetic mice, we found opposite regulation of insulin and PDX1 mRNA; insulin was decreased in diabetes, but PDX1 was increased. This suggests that elevated PDX1 mRNA levels may be insufficient to regulate insulin. In diabetic islets, PDX1 protein was localized in the cytosol, whereas in non-diabetic controls, PDX1 was in the nucleus. In contrast, overexpression of either IL-1 receptor antagonist or shuttling-deficient PDX1 restored β-cell survival and function and PDX1 nuclear localization. Our results show that nuclear localization of PDX1 is essential for a functional β-cell and provides a novel mechanism of the protective effect of IL-1 receptor antagonist on β-cell survival and function. PMID:21393239

  15. Virus-like particles that display Zika virus envelope protein domain III induce potent neutralizing immune responses in mice.

    PubMed

    Yang, Ming; Lai, Huafang; Sun, Haiyan; Chen, Qiang

    2017-08-09

    Several Zika virus (ZIKV) vaccine candidates have recently been described which use inactivated whole virus, DNA or RNA that express the virus' Envelope (E) glycoprotein as the antigen. These were successful in stimulating production of virus-targeted antibodies that protected animals against ZIKV challenges, but their use potentially will predispose vaccinated individuals to infection by the related Dengue virus (DENV). We have devised a virus like particle (VLP) carrier based on the hepatitis B core antigen (HBcAg) that displays the ZIKV E protein domain III (zDIII), and shown that it can be produced quickly and easily purified in large quantities from Nicotiana benthamiana plants. HBcAg-zDIII VLPs are shown to be highly immunogenic, as two doses elicited potent humoral and cellular responses in mice that exceed the threshold correlated with protective immunity against multiple strains of Zika virus. Notably, HBcAg-zDIII VLPs-elicited antibodies did not enhance the infection of DENV in Fc gamma receptor-expressing cells, offsetting the concern of ZIKV vaccines inducing cross-reactive antibodies and sensitizing people to subsequent DENV infection. Thus, our zDIII-based vaccine offers improved safety and lower cost production than other current alternatives, with equivalent effectiveness.

  16. Investigating the mechanism for maintaining eucalcemia despite immobility and anuria in the hibernating American black bear (Ursus americanus).

    PubMed

    Seger, Rita L; Cross, Randal A; Rosen, Clifford J; Causey, Robert C; Gundberg, Caren M; Carpenter, Thomas O; Chen, Tai C; Halteman, William A; Holick, Michael F; Jakubas, Walter J; Keisler, Duane H; Seger, Richard M; Servello, Frederick A

    2011-12-01

    Ursine hibernation uniquely combines prolonged skeletal unloading, anuria, pregnancy, lactation, protein recycling, and lipolysis. This study presents a radiographic and biochemical picture of bone metabolism in free-ranging, female American black bears (Ursus americanus) that were active (spring bears and autumn bears) or hibernating (hibernating bears). Hibernating bears included lactating and non-lactating individuals. We measured serum calcium, albumin, inorganic phosphate, creatinine, bone specific alkaline phosphatase (BSALP), CTX, parathyroid hormone, insulin-like growth factor-I (IGF-l), leptin, 25-hydroxyvitamin D [25(OH)D], 1,25-dihydroxyvitamin D [1,25(OH)(2)D] and sclerostin from 35 to 50 tranquilized hibernating bears and 14 to 35 tranquilized spring bears. We compared metacarpal cortical indices (MCI), measured by digital X-ray radiogrammetry, from 60 hunter-killed autumn bears and 79 tranquilized, hibernating bears. MCI was greater in autumn than winter in younger bears, but showed no seasonal difference in older bears. During hibernation eucalcemia was maintained, BSALP was suppressed, and CTX was in the range expected for anuria. During hibernation 1,25(OH)(2)D was produced despite anuria. 1,25(OH)(2)D and IGF-I were less in hibernating than spring bears. In a quarter of hibernating bears, sclerostin was elevated. Leptin was greater in hibernating than spring bears. In hibernating bears, leptin correlated positively with BSALP in non-lactating bears and with CTX in lactating bears. Taken together the biochemical and radiographic findings indicate that during hibernation, bone turnover was persistent, balanced, and suppressed; bone resorption was lower than expected for an unloaded skeleton; and there was no unloading-induced bone loss. The skeleton appears to perceive that it was loaded when it was actually unloaded during hibernation. However, at the level of sclerostin, the skeleton recognized that it was unloaded. During hibernation leptin

  17. Heavy Chain-Only IgG2b Llama Antibody Effects Near-Pan HIV-1 Neutralization by Recognizing a CD4-Induced Epitope That Includes Elements of Coreceptor- and CD4-Binding Sites

    PubMed Central

    Luongo, Timothy S.; Georgiev, Ivelin S.; Matz, Julie; Schmidt, Stephen D.; Louder, Mark K.; Kessler, Pascal; Yang, Yongping; McKee, Krisha; O'Dell, Sijy; Chen, Lei; Baty, Daniel; Chames, Patrick; Martin, Loïc; Mascola, John R.

    2013-01-01

    The conserved HIV-1 site of coreceptor binding is protected from antibody-directed neutralization by conformational and steric restrictions. While inaccessible to most human antibodies, the coreceptor site has been shown to be accessed by antibody fragments. In this study, we used X-ray crystallography, surface plasmon resonance, and pseudovirus neutralization to characterize the gp120-envelope glycoprotein recognition and HIV-1 neutralization of a heavy chain-only llama antibody, named JM4. We describe full-length IgG2b and IgG3 versions of JM4 that target the coreceptor-binding site and potently neutralize over 95% of circulating HIV-1 isolates. Contrary to established trends that show improved access to the coreceptor-binding region by smaller antibody fragments, the single-domain (VHH) version of JM4 neutralized less well than the full-length IgG2b version of JM4. The crystal structure at 2.1-Å resolution of VHH JM4 bound to HIV-1 YU2 gp120 stabilized in the CD4-bound state by the CD4-mimetic miniprotein, M48U1, revealed a JM4 epitope that combined regions of coreceptor recognition (including the gp120 bridging sheet, V3 loop, and β19 strand) with gp120 structural elements involved in recognition of CD4 such as the CD4-binding loop. The structure of JM4 with gp120 thus defines a novel CD4-induced site of vulnerability involving elements of both coreceptor- and CD4-binding sites. The potently neutralizing JM4 IgG2b antibody that targets this newly defined site of vulnerability adds to the expanding repertoire of broadly neutralizing antibodies that effectively neutralize HIV-1 and thereby potentially provides a new template for vaccine development and target for HIV-1 therapy. PMID:23843638

  18. Heavy chain-only IgG2b llama antibody effects near-pan HIV-1 neutralization by recognizing a CD4-induced epitope that includes elements of coreceptor- and CD4-binding sites.

    PubMed

    Acharya, Priyamvada; Luongo, Timothy S; Georgiev, Ivelin S; Matz, Julie; Schmidt, Stephen D; Louder, Mark K; Kessler, Pascal; Yang, Yongping; McKee, Krisha; O'Dell, Sijy; Chen, Lei; Baty, Daniel; Chames, Patrick; Martin, Loïc; Mascola, John R; Kwong, Peter D

    2013-09-01

    The conserved HIV-1 site of coreceptor binding is protected from antibody-directed neutralization by conformational and steric restrictions. While inaccessible to most human antibodies, the coreceptor site has been shown to be accessed by antibody fragments. In this study, we used X-ray crystallography, surface plasmon resonance, and pseudovirus neutralization to characterize the gp120-envelope glycoprotein recognition and HIV-1 neutralization of a heavy chain-only llama antibody, named JM4. We describe full-length IgG2b and IgG3 versions of JM4 that target the coreceptor-binding site and potently neutralize over 95% of circulating HIV-1 isolates. Contrary to established trends that show improved access to the coreceptor-binding region by smaller antibody fragments, the single-domain (VHH) version of JM4 neutralized less well than the full-length IgG2b version of JM4. The crystal structure at 2.1-Å resolution of VHH JM4 bound to HIV-1 YU2 gp120 stabilized in the CD4-bound state by the CD4-mimetic miniprotein, M48U1, revealed a JM4 epitope that combined regions of coreceptor recognition (including the gp120 bridging sheet, V3 loop, and β19 strand) with gp120 structural elements involved in recognition of CD4 such as the CD4-binding loop. The structure of JM4 with gp120 thus defines a novel CD4-induced site of vulnerability involving elements of both coreceptor- and CD4-binding sites. The potently neutralizing JM4 IgG2b antibody that targets this newly defined site of vulnerability adds to the expanding repertoire of broadly neutralizing antibodies that effectively neutralize HIV-1 and thereby potentially provides a new template for vaccine development and target for HIV-1 therapy.

  19. Neutralization by insulin of the hypertensive effect of dermcidin isoform 2: an environmentally induced diabetogenic and hypertensive protein.

    PubMed

    Ghosh, Rajeshwary; Bank, Sarbashri; Bhattacharya, Rabindra; Khan, Nighat N; Sinha, A Kumar

    2014-01-01

    The effect of dermcidin isoform 2 (dermcidin), an environmentally induced stress protein, was investigated on the genesis of diabetes mellitus and hypertension, the two major atherosclerotic risk factors. The role of dermcidin as an atherosclerotic risk factor related to the impaired systemic insulin level was investigated. Dermcidin was prepared by electrophoresis using plasma from the subjects with acute ischemic heart disease. Injection of 0.2 μM dermcidin in mice increased the blood glucose level from 98 ± 2.45 mg/dL to 350  ± 10.2 mg/dL which was normalized by the oral administration of acetyl salicylic acid (aspirin) after 24 h. Hypertensive subjects with systolic and diastolic blood pressure of 165 mm and 95 mm of Hg, respectively, had plasma dermcidin level of 95 nM. Ingestion of acetyl salicylic acid (aspirin) (150 mg/70 kg body weight) decreased the systolic and diastolic pressures to 125 mm and 80 mm of Hg, respectively, with decrease of dermcidin level to 15 nM. Incubation of kidney cortex cells with 0.2 μM dermcidin-inhibited synthesis of (r)-cortexin, an antihypertensive protein, and the basal (r)-cortexin level was reduced from 33 nM to 15 nM. Addition of 25 μunits of insulin/mL was found to reverse the inhibition of cortexin synthesis. The effect of dermcidin as a diabetogenic and a hypertensive agent could be controlled either by aspirin or by insulin.

  20. Neutralization by Insulin of the Hypertensive Effect of Dermcidin Isoform 2: An Environmentally Induced Diabetogenic and Hypertensive Protein

    PubMed Central

    Bank, Sarbashri; Bhattacharya, Rabindra; Khan, Nighat N.; Sinha, A. Kumar

    2014-01-01

    The effect of dermcidin isoform 2 (dermcidin), an environmentally induced stress protein, was investigated on the genesis of diabetes mellitus and hypertension, the two major atherosclerotic risk factors. The role of dermcidin as an atherosclerotic risk factor related to the impaired systemic insulin level was investigated. Dermcidin was prepared by electrophoresis using plasma from the subjects with acute ischemic heart disease. Injection of 0.2 μM dermcidin in mice increased the blood glucose level from 98 ± 2.45 mg/dL to 350  ± 10.2 mg/dL which was normalized by the oral administration of acetyl salicylic acid (aspirin) after 24 h. Hypertensive subjects with systolic and diastolic blood pressure of 165 mm and 95 mm of Hg, respectively, had plasma dermcidin level of 95 nM. Ingestion of acetyl salicylic acid (aspirin) (150 mg/70 kg body weight) decreased the systolic and diastolic pressures to 125 mm and 80 mm of Hg, respectively, with decrease of dermcidin level to 15 nM. Incubation of kidney cortex cells with 0.2 μM dermcidin-inhibited synthesis of (r)-cortexin, an antihypertensive protein, and the basal (r)-cortexin level was reduced from 33 nM to 15 nM. Addition of 25 μunits of insulin/mL was found to reverse the inhibition of cortexin synthesis. The effect of dermcidin as a diabetogenic and a hypertensive agent could be controlled either by aspirin or by insulin. PMID:24649391

  1. Synonymous Deoptimization of Foot-and-Mouth Disease Virus Causes Attenuation In Vivo while Inducing a Strong Neutralizing Antibody Response

    PubMed Central

    Diaz-San Segundo, Fayna; Medina, Gisselle N.; Ramirez-Medina, Elizabeth; Velazquez-Salinas, Lauro; Koster, Marla; Grubman, Marvin J.

    2015-01-01

    ABSTRACT Codon bias deoptimization has been previously used to successfully attenuate human pathogens, including poliovirus, respiratory syncytial virus, and influenza virus. We have applied a similar technology to deoptimize the capsid-coding region (P1) of foot-and-mouth disease virus (FMDV). Despite the introduction of 489 nucleotide changes (19%), synonymous deoptimization of the P1 region rendered a viable FMDV progeny. The resulting strain was stable and reached cell culture titers similar to those obtained for wild-type (WT) virus, but at reduced specific infectivity. Studies in mice showed that 100% of animals inoculated with the FMDV A12 P1 deoptimized mutant (A12-P1 deopt) survived, even when the animals were infected at doses 100 times higher than the dose required to cause death by WT virus. All mice inoculated with the A12-P1 deopt mutant developed a strong antibody response and were protected against subsequent lethal challenge with WT virus at 21 days postinoculation. Remarkably, the vaccine safety margin was at least 1,000-fold higher for A12-P1 deopt than for WT virus. Similar patterns of attenuation were observed in swine, in which animals inoculated with A12-P1 deopt virus did not develop clinical disease until doses reached 1,000 to 10,000 times the dose required to cause severe disease in 2 days with WT A12. Consistently, high levels of antibody titers were induced, even at the lowest dose tested. These results highlight the potential use of synonymous codon pair deoptimization as a strategy to safely attenuate FMDV and further develop live attenuated vaccine candidates to control such a feared livestock disease. IMPORTANCE Foot-and-mouth disease (FMD) is one of the most feared viral diseases that can affect livestock. Although this disease appeared to be contained in developed nations by the end of the last century, recent outbreaks in Europe, Japan, Taiwan, South Korea, etc., have demonstrated that infection can spread rapidly, causing

  2. Neutral ligand induced methane elimination from rare-earth metal tetramethylaluminates up to the six-coordinate carbide state.

    PubMed

    Venugopal, Ajay; Kamps, Ina; Bojer, Daniel; Berger, Raphael J F; Mix, Andreas; Willner, Alexander; Neumann, Beate; Stammler, Hans-Georg; Mitzel, Norbert W

    2009-08-07

    The reaction of 1,3,5-trimethyl-1,3,5-triazacyclohexane (TMTAC) with [La{Al(CH(3))(4)}(3)] resulted in C-H activation, leading to the formation of [(TMTAC)La{Al(CH(3))(4)}{(mu(3)-CH(2))[Al(CH(3))(2)(mu(2)-CH(3))](2)}] (1) containing a bis(aluminate) dianion and subsequent extrusion of methane. A similar reaction with [Y{Al(CH(3))(4)}(3)] led to the formation of CH(4), [TMTAC{Al(CH(3))(3)}(2)] (2) and {[(TMTAC)Y][Y(2)(mu(2)-CH(3))][{(mu(6)-C)[Al(mu(2)-CH(3))(2)(CH(3))](3)}{(mu(3)-CH(2))(mu(2)-CH(3))Al(CH(3))(2)}(2)] (3), containing a six-coordinate carbide ion and two [CH(2)Al(CH(3))(3)](2)(-) anions. Compound 3 is a product of multiple C-H activation. This reaction was monitored by in situ(1)H NMR spectroscopy. The analogous reaction with [Sm{Al(CH(3))(4)}(3)] led to the formation of 2, of [(TMTAC)Sm{(mu(2)-CH(3))(CH(3))(2)Al}(2){(mu(3)-CH(2))(2)Al(CH(3))(2)}(2)] (4), which contains a tris(aluminate) trianion, and [{(TMTAC)Sm}{Sm(2)(mu(2)-CH(3))}{(mu(6)-C)[Al(mu(2)-CH(3))(2)(CH(3))](3)}{(mu(3)-CH(2))(mu(2)-CH(3))Al(CH(3))(2)}(2)] (5), which is isostructural to 3. The products were characterised by elemental analyses (except 4, 5), 1 by multinuclear NMR spectroscopy and compounds 1, 2, 3, 4 and 5 by X-ray crystallography. Quantumchemical calculations were undertaken to support the crystallographic data analysis and confirm the structure of 3 and to compare it with an analogous compound where the central six-coordinate carbon has been replaced by oxygen. The investigations point to a mechanism of sterically induced condensation of [Al(CH(3))(4)](-) groups in close proximity in the coordination spheres of the rare-earth metal atoms, which is dependent on the size of these metal atoms.

  3. Neutralizing Antibodies Induced by Recombinant Virus-Like Particles of Enterovirus 71 Genotype C4 Inhibit Infection at Pre- and Post-attachment Steps

    PubMed Central

    Ku, Zhiqiang; Ye, Xiaohua; Huang, Xulin; Cai, Yicun; Liu, Qingwei; Li, Yan; Su, Zhiguo; Huang, Zhong

    2013-01-01

    Background Enterovirus 71 (EV71) is a major causative agent of hand, foot and mouth disease, which has been prevalent in Asia–Pacific regions, causing significant morbidity and mortality in young children. Antibodies elicited by experimental EV71 vaccines could neutralize infection in vitro and passively protect animal models from lethal challenge, indicating that neutralizing antibodies play an essential role in protection. However, how neutralizing antibodies inhibit infection in vitro remains unclear. Methods/Findings In the present study, we explored the mechanisms of neutralization by antibodies against EV71 virus-like particles (VLPs). Recombinant VLPs of EV71 genotype C4 were produced in insect cells using baculovirus vectors. Immunization with the VLPs elicited a high-titer, EV71-specific antibody response in mice. Anti-VLP mouse sera potently neutralized EV71 infection in vitro. The neutralizing antibodies in the anti-VLP mouse sera were found to target mainly an extremely conserved epitope (FGEHKQEKDLEYGAC) located at the GH loop of the VP1 protein. The neutralizing anti-VLP antisera were able to inhibit virus binding to target cells efficiently. In addition, post-attachment treatment of virus-bound cells with the anti-VLP antisera also neutralized virus infection, although the antibody concentration required was higher than that of the pre-attachment treatment. Conclusions Collectively, our findings represent a valuable addition to the understanding of mechanisms of EV71 neutralization and have strong implications for EV71 vaccine development. PMID:23451250

  4. Recombinant Receptor-Binding Domains of Multiple Middle East Respiratory Syndrome Coronaviruses (MERS-CoVs) Induce Cross-Neutralizing Antibodies against Divergent Human and Camel MERS-CoVs and Antibody Escape Mutants.

    PubMed

    Tai, Wanbo; Wang, Yufei; Fett, Craig A; Zhao, Guangyu; Li, Fang; Perlman, Stanley; Jiang, Shibo; Zhou, Yusen; Du, Lanying

    2017-01-01

    Middle East respiratory syndrome coronavirus (MERS-CoV) binds to cellular receptor dipeptidyl peptidase 4 (DPP4) via the spike (S) protein receptor-binding domain (RBD). The RBD contains critical neutralizing epitopes and serves as an important vaccine target. Since RBD mutations occur in different MERS-CoV isolates and antibody escape mutants, cross-neutralization of divergent MERS-CoV strains by RBD-induced antibodies remains unknown. Here, we constructed four recombinant RBD (rRBD) proteins with single or multiple mutations detected in representative human MERS-CoV strains from the 2012, 2013, 2014, and 2015 outbreaks, respectively, and one rRBD protein with multiple changes derived from camel MERS-CoV strains. Like the RBD of prototype EMC2012 (EMC-RBD), all five RBDs maintained good antigenicity and functionality, the ability to bind RBD-specific neutralizing monoclonal antibodies (MAbs) and the DPP4 receptor, and high immunogenicity, able to elicit S-specific antibodies. They induced potent neutralizing antibodies cross-neutralizing 17 MERS pseudoviruses expressing S proteins of representative human and camel MERS-CoV strains identified during the 2012-2015 outbreaks, 5 MAb escape MERS-CoV mutants, and 2 live human MERS-CoV strains. We then constructed two RBDs mutated in multiple key residues in the receptor-binding motif (RBM) of RBD and demonstrated their strong cross-reactivity with anti-EMC-RBD antibodies. These RBD mutants with diminished DPP4 binding also led to virus attenuation, suggesting that immunoevasion after RBD immunization is accompanied by loss of viral fitness. Therefore, this study demonstrates that MERS-CoV RBD is an important vaccine target able to induce highly potent and broad-spectrum neutralizing antibodies against infection by divergent circulating human and camel MERS-CoV strains.

  5. An equine herpesvirus type 1 (EHV-1) vector expressing Rift Valley fever virus (RVFV) Gn and Gc induces neutralizing antibodies in sheep.

    PubMed

    Said, Abdelrahman; Elmanzalawy, Mona; Ma, Guanggang; Damiani, Armando Mario; Osterrieder, Nikolaus

    2017-08-14

    Rift Valley fever virus (RVFV) is an arthropod-borne bunyavirus that can cause serious and fatal disease in humans and animals. RVFV is a negative-sense RNA virus of the Phlebovirus genus in the Bunyaviridae family. The main envelope RVFV glycoproteins, Gn and Gc, are encoded on the M segment of RVFV and known inducers of protective immunity. In an attempt to develop a safe and efficacious RVF vaccine, we constructed and tested a vectored equine herpesvirus type 1 (EHV-1) vaccine that expresses RVFV Gn and Gc. The Gn and Gc genes were custom-synthesized after codon optimization and inserted into EHV-1 strain RacH genome. The rH_Gn-Gc recombinant virus grew in cultured cells with kinetics that were comparable to those of the parental virus and stably expressed Gn and Gc. Upon immunization of sheep, the natural host, neutralizing antibodies against RVFV were elicited by rH_Gn-Gc and protective titers reached to 1:320 at day 49 post immunization but not by parental EHV-1, indicating that EHV-1 is a promising vector alternative in the development of a safe marker RVFV vaccine.

  6. ArI/ArII laser induced fluorescence system for measurement of neutral and ion dynamics in a large scale helicon plasma

    NASA Astrophysics Data System (ADS)

    Kelly, R. F.; Meaney, K. D.; Gilmore, M.; Desjardins, T. R.; Zhang, Y.

    2016-11-01

    In order to investigate the role of both neutral and ion dynamics in large-scale helicon discharges, a laser induced fluorescence (LIF) system capable of measuring both ArI and ArII fluorescence using a single tunable laser is being developed. The system is based on a >250 mW solid state laser. For ArI measurements, the laser pumps the metastable (2P03/2)4s level to the (2P01/2)4p level using 696.7352 nm light, and fluorescence radiation from decay to the (2P01/2)4s level at 772.6333 nm is observed. For ArII, currently in development, the metastable (3P)3d 4F7/2 level will be pumped to the (3P)4p 4D07/2 level using 688.8511 nm, and decay fluorescence to the (3P)4s 4P5/2 level at 434.9285 nm measured. Here all wavelengths are in a vacuum.

  7. Neutralizing circulating ghrelin by expressing a growth hormone secretagogue receptor-based protein protects against high-fat diet-induced obesity in mice.

    PubMed

    Gagnon, J; Zhu, L; Anini, Y; Wang, Q

    2015-09-01

    Ghrelin is a stomach-derived peptide hormone that stimulates appetite and promotes adiposity through binding to the growth hormone secretagogue receptor (GHS-R1a). Administration of ghrelin in rodents increases weight gain due to stimulating food intake and reducing fat utilization. Therefore, reducing circulating ghrelin levels holds the potential to reduce weight gain. We developed a GHS-R1a-fusion constructs of a decoy protein containing the ligand-binding domains of the ghrelin receptor. Intramuscular injection of the GHSR/Fc plasmid decreased circulating levels of acylated-ghrelin. When challenged with the high fat diet, treated mice displayed reduced weight gain compared with controls, which was associated with reduced fat accumulation in the peritoneum but not lean mass. Quantitative PCR with reverse transcription showed increased PPARγ and hormone sensitive lipase transcripts levels in adipose tissue of treated animals, illustrating a preference for increased fat utilization. Intra-peritoneal glucose tolerance and insulin tolerance tests showed improved glucose clearance and insulin sensitivity in GHSR/Fc treated animals. We suggest that in vivo expression of the GHSR-based fusion protein prevents diet-induced weight gain, altering adipose gene expression and improving glucose tolerance. These findings, while confirming the role of ghrelin in peripheral energy metabolism, suggest that a strategy involving neutralization of the circulation ghrelin by intramuscular injection of the GHSR1/Fc fusion construct may find clinical application in the treatment of obesity.

  8. Ammonia-containing industrial effluents, lethal to rainbow trout, induce vacuolisation and Neutral Red uptake in the rainbow trout gill cell line, RTgill-W1.

    PubMed

    Dayeh, Vivian R; Schirmer, Kristin; Bols, Niels C

    2009-02-01

    Nine samples of whole effluent from the operation of an industrial plant over the course of one year, were tested on rainbow trout for lethality and on the rainbow trout gill cell line, RTgill-W1, for metabolic activity, plasma membrane integrity, and lysosomal activity, as measured by using the alamar Blue (AB), 5-carboxyfluorescein diacetate acetoxymethyl (CFDA-AM), and neutral red (NR) assays, respectively. None of the nine samples caused a loss of plasma membrane integrity, and only two caused a transitory decline in metabolism. Three samples caused massive vacuolisation in RTgill-W1 cells, which was accompanied by increased uptake of NR, and only these three samples were lethal to the rainbow trout. The addition of ammonia to RTgill-W1 cultures also induced vacuolisation and NR uptake, with little change in plasma membrane integrity or metabolism. Subsequently, the effluent source was identified as a nitrogen product producer, and variable levels of ammonia were found in the nine samples. Three of the four samples with the highest non-ionised ammonia levels were those which were toxic to rainbow trout and which caused vacuoles in RTgill-W1 cells. The close correlation between rainbow trout-killing and RTgill-W1 vacuolisation by the effluents, suggests that vacuolisation of RTgill-W1 cells could be used to indicate effluents which would be toxic to rainbow trout as a result of their ammonia content.

  9. The fission yeast mitotic activator cdc25 and sucrose induce early flowering synergistically in the day-neutral Nicotiana tabacum cv. Samsun.

    PubMed

    Teichmanová, Martina; Masková, Petra; Vojvodová, Petra; Krekule, Jan; Francis, Dennis; Lipavská, Helena

    2007-01-01

    Here, the tobacco (Nicotiana tabacum) day-neutral (DN) cv. Samsun transformed with the Schizosaccharomyces pombe mitotic activator gene Spcdc25 was used to study the onset of flowering. Wild type (WT) and cdc25 plants were grown from seeds in vitro until they were 20 cm high. Apical and basal nodes were then subcultured repeatedly and the regenerated plants were used to document time to flowering and the number of leaves formed before flowering. Three sucrose treatments (3, 5 or 7% (weight/volume)) were used and measurements of leaf endogenous soluble carbohydrates were performed. In the 3% treatment, cdc25 plants flowered but WT plants did not. The higher sucrose treatments enabled WT flowering; two-thirds of the plants flowered at 5%, while all plants flowered at 7% sucrose. However, in all treatments, cdc25 plants exhibited significantly earlier flowering and fewer leaves compared with wild type. Remarkably, a typical acropetal flowering gradient in WT plants did not occur in cdc25 plants. In cdc25 leaves, there were significantly higher amounts of endogenous sugars with a higher proportion of sucrose compared with WT. Our data demonstrate that Spcdc25 expression and sucrose act synergistically to induce precocious flowering.

  10. SARS-CoV spike proteins expressed by the vaccinia virus Tiantan strain: secreted sq protein induces robust neutralization antibody in mice.

    PubMed

    Yan, Kexia; Tan, Wenjie; Wang, Huijuan; Wang, Yue; Zhang, Xiangmin; Li, Yan; Ruan, Li

    2009-02-01

    The spike (S) glycoprotein of severe acute respiratory syndrome coronavirus (SARS-CoV) is a major target in the development of diagnostic assays and vaccines, but its antigenic and immunogenic properties remain unclear. Seven SARS-CoV spike proteins (S, SQ, S1, RBD, S2, S2Q, and CX) were generated using the modified vaccinia virus (Tiantan strain) as a vector, and their antigenicity and immunogenicity were evaluated. The secreted SQ protein in which the transmembrane domain was deleted, as well as the full-length spike protein, showed the most potential to induce the production of neutralizing antibody (nAb) in mice. S1 and RBD proteins initialized significantly lower levels of nAb production. In addition, the S proteins were recognized specifically by the sera of convalescent patients with SARS, and that of mice immunized with inactivated SARS-CoV, but did not react with anti-sera of HCoV-OC43 or HCoV-229E, or sera from healthy donors (although RBD showed a false-positive in 1 of 55 control samples of human sera). Our results demonstrate that SQ protein may be an effective vaccine candidate and a convenient and safe diagnostic antigen for SARS-CoV.

  11. A transcriptomic analysis of American lobster (Homarus americanus) immune response during infection with the bumper car parasite Anophryoides haemophila.

    PubMed

    Clark, K Fraser; Acorn, Adam R; Greenwood, Spencer J

    2013-06-01

    Anophryoides haemophila is an important protistan parasite of American lobster, Homarus americanus, as it has been found to infect lobsters in the wild as well as causing major losses of lobsters maintained in commercial holding facilities. Expression of over 14,500 H. americanus hepatopancreatic genes were monitored during an A. haemophila infection challenge in order to elucidate molecular mechanisms involved in the lobster immune response. One hundred and forty-five genes were found to be differentially expressed during infection. For many genes, this study is the first to link their expression to an immune response to a known lobster pathogen. Several of the genes have previously been linked to crustacean or invertebrate immune response including: several anti-lipopolysaccharide factor isoforms (ALFHa), acute phase serum amyloid protein A (SAA), a serine protease inhibitor, a toll-like receptor, several haemocyanin subunits, phagocyte signaling-impaired protein, vitelline membrane outer layer protein-1, trypsin, and a C-type lectin receptor. Microarray results were verified using RT-qPCR and agreement was good between the two methods. The expression of six ALFHa isoforms was monitored via microarray where ALFHa-1, ALFHa-2, ALFHa-4 and ALFHa-6 were differentially expressed while ALFHa-3 and ALFHa7 were not. RT-qPCR analysis confirmed that ALFHa-1, ALFHA-2 and ALFHa-4 expression increased during infection with a peak at 5-7weeks for ALFHa-1 and 10weeks for ALFHa-2 and ALFHa-4. This suggests that different ALFHa isoforms are temporally expressed during A. haemophila infection. Importantly, these results provide evidence that different ALFHa isoforms have more significant roles in responding to A. haemophila infection. Significant increases in SAA gene expression were also found, corroborating previous findings of increased SAA expression during Aerococcus viridans infections; highlighting the importance of SAA as a marker of H. americanus immune activation and

  12. Purification and amino acid composition of a peptide with molt-inhibiting activity from the lobster, Homarus americanus.

    PubMed

    Chang, E S; Bruce, M J; Newcomb, R W

    1987-01-01

    A peptide was isolated and purified from sinus glands of the lobster, Homarus americanus, that was able to decrease circulating titers of ecdysteroids and increase the molt interval of eyestalk-ablated juvenile lobsters. This molt-inhibiting activity was demonstrated to consist of two very closely related peptides by means of high-performance liquid chromatography and gel electrophoresis. By means of amino acid analyses, a molecular weight of approximately 8700 was obtained.

  13. Photodetachment process for beam neutralization

    DOEpatents

    Fink, Joel H. [Livermore, CA; Frank, Alan M. [Livermore, CA

    1979-02-20

    A process for neutralization of accelerated ions employing photo-induced charge detachment. The process involves directing a laser beam across the path of a negative ion beam such as to effect photodetachment of electrons from the beam ions. The frequency of the laser beam employed is selected to provide the maximum cross-section for the photodetachment process.

  14. Photodetachment process for beam neutralization

    DOEpatents

    Fink, J.H.; Frank, A.M.

    1979-02-20

    A process for neutralization of accelerated ions employing photo-induced charge detachment is disclosed. The process involves directing a laser beam across the path of a negative ion beam such as to effect photodetachment of electrons from the beam ions. The frequency of the laser beam employed is selected to provide the maximum cross-section for the photodetachment process. 2 figs.

  15. On the population development of the introduced razor clam Ensis americanus near the island of Sylt (North Sea)

    NASA Astrophysics Data System (ADS)

    Armonies, W.; Reise, K.

    1998-09-01

    The American razor clam Ensis americanus (= E. directus) was introduced into the eastern North Sea in the late 1970s. By larval and poslarval drifting the species rapidly extended its distribution, now ranging from the English Channel to the Kattegat. Near the island of Sylt in the eastern North Sea it has been recorded since 1979. Recruitment was rather irregular, with about six strong year-classes within two decades. Growth seems comparable with populations in its native range (Atlantic North America). Although present in the lower intertidal zone, maximum densities occurred in shallow subtidal sand with a biomass similar to that of dense beds of native cockles and mussels in the adjacent intertidal zone. Ensis americanus established in otherwise sparsely faunated sand (channels exposed to strong currents) as well as in dense infaunal assemblages (lower intertidal and subtidally). There were no significant interactions with resident species. In dense beds of razor clams, however, fine sediment particles accumulated which may have altered abundances of polychaetes. In spite of high annual variability, E. americanus has become a prominent component of the macrobenthos in shallow subtidal sands of the North Sea.

  16. First report of Taenia arctos (Cestoda: Taeniidae) from grizzly (Ursus arctos horribilis) and black bears (Ursus americanus) in North America.

    PubMed

    Catalano, Stefano; Lejeune, Manigandan; Verocai, Guilherme G; Duignan, Pádraig J

    2014-04-01

    The cestode Taenia arctos was found at necropsy in the small intestine of a grizzly (Ursus arctos horribilis) and a black bear (Ursus americanus) from Kananaskis Country in southwestern Alberta, Canada. The autolysis of the tapeworm specimens precluded detailed morphological characterization of the parasites but molecular analysis based on mitochondrial DNA cytochrome c oxidase subunit 1 gene confirmed their identity as T. arctos. This is the first report of T. arctos from definitive hosts in North America. Its detection in Canadian grizzly and black bears further supports the Holarctic distribution of this tapeworm species and its specificity for ursids as final hosts. Previously, T. arctos was unambiguously described at its adult stage in brown bears (Ursus arctos arctos) from Finland, and as larval stages in Eurasian elk (Alces alces) from Finland and moose (Alces americanus) from Alaska, USA. Given the morphological similarity between T. arctos and other Taenia species, the present study underlines the potential for misidentification of tapeworm taxa in previous parasitological reports from bears and moose across North America. The biogeographical history of both definitive and intermediate hosts in the Holarctic suggests an ancient interaction between U. arctos, Alces spp., and T. arctos, and a relatively recent host-switching event in U. americanus.

  17. Anti-lipopolysaccharide factors in the American lobster Homarus americanus: Molecular characterization and transcriptional response to Vibrio fluvialis challenge

    PubMed Central

    Beale, K.M.; Towle, D.W.; Jayasundara, N.; Smith, C.M.; Shields, J.D.; Small, H.J.; Greenwood, S.J.

    2008-01-01

    Two partial mRNA sequences predicted to encode anti-lipopolysaccharide factors (ALFs) were identified among expressed sequence tags generated from the American lobster Homarus americanus and complete cDNA sequences were obtained from library clones. Comparison of the translated amino acid sequences to those publicly available confirmed similarity to arthropod anti-lipopolysaccharide factors. Both protein sequences, designated ALFHa-1 and ALFHa-2, contained an N-terminal signal peptide and two half-cysteines participating in a disulfide bridge, features conserved in other ALFs. Predicted secondary structures were similar to that described for the ALF from the horseshoe crab Limulus polyphemus. As part of an exploratory study of immunity in H. americanus, lobsters were injected with the bacterium Vibrio fluvialis and gill, hematopoietic, and hepatopancreas tissues were sampled for analysis of gene expression of ALFHa-1 and ALFHa-2 by quantitative PCR. The relative abundance of ALFHa-2 mRNA was not significantly affected by Vibrio injection in any of the three tissues tested. In contrast, ALFHa-1 mRNA levels in gills were increased by the treatment some 17-fold. Our results support a molecularly specific regulation of antimicrobial proteins in response to bacterial infection in H. americanus. PMID:19956341

  18. Immunization with a soluble CD4-gp120 complex preferentially induces neutralizing anti-human immunodeficiency virus type 1 antibodies directed to conformation-dependent epitopes of gp120.

    PubMed

    Kang, C Y; Hariharan, K; Nara, P L; Sodroski, J; Moore, J P

    1994-09-01

    Preservation of the conformation of recombinant gp120 in an adjuvant, enabling it to elicit conformation-dependent, epitope-specific, broadly neutralizing antibodies, may be critical for the development of any gp120-based human immunodeficiency virus type 1 (HIV-1) vaccine. It was hypothesized that recombinant gp120 complexed with recombinant CD4 could stabilize the conformation-dependent neutralizing epitopes and effectively deliver them to the immune system. Therefore, a soluble CD4-gp120 complex in Syntex adjuvant formulation was tested with mice for its ability to induce neutralizing anti-gp120 antibody responses. Seventeen monoclonal antibodies (MAbs) were generated and characterized. Immunochemical studies, neutralization assays, and mapping studies with gp120 mutants indicated that the 17 MAbs fell into three groups. Four of them were directed to what is probably a conformational epitope involving the C1 domain and did not possess virus-neutralizing activities. Another four MAbs bound to V3 peptide 302-321 and exhibited cross-reactive gp120 binding and relatively weak virus-neutralizing activities. These MAbs were very sensitive to amino acid substitutions, not only in the V3 regions but also in the base of the V1/V2 loop, implying a conformational constraint on the epitope. The last group of nine MAbs recognized conformation-dependent epitopes near the CD4 binding site of gp120 and inhibited the gp120-soluble CD4 interaction. Four of these nine MAbs showed broadly neutralizing activities against multiple laboratory-adapted strains of HIV-1, three of them neutralized only HIVIIIB, and the two lower-affinity MAbs did not neutralize any strain tested. Collectively, the results from this study indicate that immunization with the CD4-gp120 complex can elicit antibodies to conformationally sensitive gp120 epitopes, with some of the antibodies having broadly neutralizing activities. We suggest that immunization with CD4-gp120 complexes may be worth evaluating

  19. Ultracold neutral plasmas

    NASA Astrophysics Data System (ADS)

    Lyon, M.; Rolston, S. L.

    2017-01-01

    By photoionizing samples of laser-cooled atoms with laser light tuned just above the ionization limit, plasmas can be created with electron and ion temperatures below 10 K. These ultracold neutral plasmas have extended the temperature bounds of plasma physics by two orders of magnitude. Table-top experiments, using many of the tools from atomic physics, allow for the study of plasma phenomena in this new regime with independent control over the density and temperature of the plasma through the excitation process. Characteristic of these systems is an inhomogeneous density profile, inherited from the density distribution of the laser-cooled neutral atom sample. Most work has dealt with unconfined plasmas in vacuum, which expand outward at velocities of order 100 m/s, governed by electron pressure, and with lifetimes of order 100 μs, limited by stray electric fields. Using detection of charged particles and optical detection techniques, a wide variety of properties and phenomena have been observed, including expansion dynamics, collective excitations in both the electrons and ions, and collisional properties. Through three-body recombination collisions, the plasmas rapidly form Rydberg atoms, and clouds of cold Rydberg atoms have been observed to spontaneously avalanche ionize to form plasmas. Of particular interest is the possibility of the formation of strongly coupled plasmas, where Coulomb forces dominate thermal motion and correlations become important. The strongest impediment to strong coupling is disorder-induced heating, a process in which Coulomb energy from an initially disordered sample is converted into thermal energy. This restricts electrons to a weakly coupled regime and leaves the ions barely within the strongly coupled regime. This review will give an overview of the field of ultracold neutral plasmas, from its inception in 1999 to current work, including efforts to increase strong coupling and effects on plasma properties due to strong coupling.

  20. Differential modulation of hippocampal chemical-induced injury response by ebselen, pentoxifylline, and TNFalpha-, IL-1alpha-, and IL-6-neutralizing antibodies.

    PubMed

    Jean Harry, G; Bruccoleri, Alessandra; Lefebvre d'Hellencourt, Christian

    2003-08-15

    The proinflammatory cytokines tumor necrosis factor (TNFalpha), interleukin-1 (IL-1alpha), and interleukin-6 (IL-6) have been associated with various models of hippocampal damage. To examine their role in initiation of an acute hippocampal injury response, 21-day-old male CD-1 mice received an acute intraperitoneal (i.p.) injection of trimethyltin hydroxide (TMT; 2.0 mg/kg) to produce necrosis of dentate granule neurons, astrocyte, and microglia reactivity. Tremors and intermittent seizures were evident at 24 hr. Intercellular adhesion molecule-1 (ICAM-1), glial fibrillary acidic protein (GFAP), anti-apoptotic TNFalpha-inducible early response gene (A-20), macrophage inflammatory protein (MIP)-1alpha, TNFalpha, IL-1alpha, IL-6, and caspase 3 mRNA levels were significantly elevated. Pretreatment with the antioxidant, ebselen, decreased ICAM-1, A-20, and TNFbeta elevations. Pentoxifylline blocked elevations in A-20 and decreased elevations in GFAP mRNA levels. Neither prevented histopathology or behavioral effects. Intracisternal injection of TNFalpha-neutralizing antibody significantly inhibited both behavioral effects and histopathology. RNase protection assays showed that TMT-induced elevations in mRNA levels for ICAM-1, A-20, GFAP, MIP-1alpha, IL-1alpha, TNFalpha, TNFbeta, and caspase 3 were blocked by anti-TNFalpha. These data demonstrate a significant role for TNFalpha in an acute neuro-injury in the absence of contribution from infiltrating cells. The cerebellum shows limited if any damage after TMT; however, in combination with the i.c.v. injection, elevations were seen in GFAP and in EB-22, a murine acute-phase response gene homologous to the alpha (1)-antichymotrypsin gene. Elevations were similar for artificial cerebral spinal fluid and anti-IL-1alpha, and significantly increased with anti-TNFalpha, anti-IL-6, or the combination of antibodies. Responses seen in the cerebellum suggest synergistic interactions between the baseline state of the cell and

  1. Effects of SA7060, a novel dual inhibitor of neutral endopeptidase and angiotensin-converting enzyme, on deoxycorticosterone acetate-salt-induced hypertension in rats.

    PubMed

    Kuro, T; Okahara, A; Nose, M; Ikuse, T; Matsumura, Y

    2000-07-01

    We evaluated whether a novel dual inhibitor of neutral endopeptidase (NEP) and angiotensin-converting enzyme (ACE), SA7060, (S)-2-[3-[(S)-2-(butoxycarbonyl)-2-hydroxyethyl]-3-isobutylureido] -3-(2-naphtyl) propionic acid, prevents deoxycorticosterone acetate (DOCA)-salt-induced hypertension and related organ damage, such as cardiovascular hypertrophy, renal dysfunction and renal tissue injury in rats. The effectiveness was compared with candoxatril and enalapril, which are a selective NEP and ACE inhibitor, respectively. During DOCA-salt treatment for 4 weeks, the rats were given SA7060, candoxatril, enalapril or vehicle, once daily by gavage. The 4-weeks treatment with DOCA and salt produced progressive increases in systolic blood pressure. Daily administration of SA7060, candoxatril or enalapril significantly suppressed the development of hypertension induced by DOCA and salt, although the effect of enalapril was less potent at 4-weeks of the treatment period. In vehicle-treated DOCA-salt rats, decreases in creatinine clearance and increases in urinary excretion of protein and blood urea nitrogen were observed. This functional damage was improved most efficiently by the treatment with SA7060. There were significant increases in urinary excretions of atrial natriuretic peptide and cyclic GMP in SA7060- or candoxatril-treated animals. Histopathological examination of the kidney in DOCA-salt rats revealed tubular, glomerular and vascular lesions, all of which were improved in animals given SA7060 or candoxatril. When the vascular hypertrophy of the aorta was evaluated, there were significant increases in wall thickness, wall area and the wall-to-lumen ratio in vehicle-treated DOCA-salt rats compared with the sham rats. The development of vascular hypertrophy was suppressed by the treatment with SA7060, candoxatril or enalapril. Our findings indicate that SA7060 efficiently prevents DOCA-salt-induced hypertension and related tissue injury, mainly by inhibiting NEP

  2. Dengue Type Four Viruses with E-Glu345Lys Adaptive Mutation from MRC-5 Cells Induce Low Viremia but Elicit Potent Neutralizing Antibodies in Rhesus Monkeys

    PubMed Central

    Li, Xiao-Feng; Tsai, Meng-Ju; Hsiao, Hung-Ju; Peng, Jia-Guan; Sue, Shih-Che; Qin, Cheng-Feng; Wu, Suh-Chin

    2014-01-01

    Knowledge of virulence and immunogenicity is important for development of live-attenuated dengue vaccines. We previously reported that an infectious clone-derived dengue type 4 virus (DENV-4) passaged in MRC-5 cells acquired a Glu345Lys (E-E345K) substitution in the E protein domain III (E-DIII). The same cloned DENV-4 was found to yield a single E-Glu327Gly (E-E327G) mutation after passage in FRhL cells and cause the loss of immunogenicity in rhesus monkeys. Here, we used site-directed mutagenesis to generate the E-E345K and E-E327G mutants from DENV-4 and DENV-4Δ30 infectious clones and propagated in Vero or MRC-5 cells. The E-E345K mutations were consistently presented in viruses recovered from MRC-5 cells, but not Vero cells. Recombinant E-DIII proteins of E345K and E327G increased heparin binding correlated with the reduced infectivity by heparin treatment in cell cultures. Different from the E-E327G mutant viruses to lose the immunogencity in rhesus monkeys, the E-E345K mutant viruses were able to induce neutralizing antibodies in rhesus monkeys with an almost a 10-fold lower level of viremia as compared to the wild type virus. Monkeys immunized with the E-E345K mutant virus were completely protected with no detectable viremia after live virus challenges with the wild type DENV-4. These results suggest that the E-E345K mutant virus propagated in MRC-5 cells may have potential for the use in live-attenuated DENV vaccine development. PMID:24959738

  3. Metabolic side effects induced by olanzapine treatment are neutralized by CB1 receptor antagonist compounds co-administration in female rats.

    PubMed

    Lazzari, P; Serra, V; Marcello, S; Pira, M; Mastinu, A

    2017-07-01

    Weight gain is an important side effect of most atypical antipsychotic drugs such as olanzapine. Moreover, although many animal models with metabolic side effects have been well defined, the interaction with other pathways has to be considered. The endocannabinoid system and the CB1 receptor (CB1R) are among the most promising central and peripheral targets involved in weight and energy balance. In this study we developed a rat model based 15-days treatment with olanzapine that shows weight gain and an alteration of the blood parameters involved in the regulation of energy balance and glucose metabolism. Consequently, we analysed whether, and by which mechanism, a co-treatment with the novel CB1R neutral antagonist NESS06SM, could attenuate the adverse metabolic effects of olanzapine compared to the reference CB1R inverse agonist rimonabant. Our results showed alterations of the cannabinoid markers in the nucleus accumbens and of orexigenic/anorexigenic markers in the hypothalamus of female rats treated with olanzapine. These molecular modifications could explain the excessive food intake and the resulting weight gain. Moreover, we confirmed that a co-treatment with CB1R antagonist/inverse agonist compounds decreased food intake and weight increment and restored all blood parameters, without altering the positive effects of olanzapine on behaviour. Furthermore, rimonabant and NESS06SM restored the metabolic enzymes in the liver and fat tissue altered by olanzapine. Therefore, CB1 receptor antagonist/inverse agonist compounds could be good candidate agents for the treatment of weight gain induced by olanzapine. Copyright © 2017 Elsevier B.V. and ECNP. All rights reserved.

  4. Dengue type four viruses with E-Glu345Lys adaptive mutation from MRC-5 cells induce low viremia but elicit potent neutralizing antibodies in rhesus monkeys.

    PubMed

    Lin, Hsiao-Han; Lee, Hsiang-Chi; Li, Xiao-Feng; Tsai, Meng-Ju; Hsiao, Hung-Ju; Peng, Jia-Guan; Sue, Shih-Che; Qin, Cheng-Feng; Wu, Suh-Chin

    2014-01-01

    Knowledge of virulence and immunogenicity is important for development of live-attenuated dengue vaccines. We previously reported that an infectious clone-derived dengue type 4 virus (DENV-4) passaged in MRC-5 cells acquired a Glu345Lys (E-E345K) substitution in the E protein domain III (E-DIII). The same cloned DENV-4 was found to yield a single E-Glu327Gly (E-E327G) mutation after passage in FRhL cells and cause the loss of immunogenicity in rhesus monkeys. Here, we used site-directed mutagenesis to generate the E-E345K and E-E327G mutants from DENV-4 and DENV-4Δ30 infectious clones and propagated in Vero or MRC-5 cells. The E-E345K mutations were consistently presented in viruses recovered from MRC-5 cells, but not Vero cells. Recombinant E-DIII proteins of E345K and E327G increased heparin binding correlated with the reduced infectivity by heparin treatment in cell cultures. Different from the E-E327G mutant viruses to lose the immunogencity in rhesus monkeys, the E-E345K mutant viruses were able to induce neutralizing antibodies in rhesus monkeys with an almost a 10-fold lower level of viremia as compared to the wild type virus. Monkeys immunized with the E-E345K mutant virus were completely protected with no detectable viremia after live virus challenges with the wild type DENV-4. These results suggest that the E-E345K mutant virus propagated in MRC-5 cells may have potential for the use in live-attenuated DENV vaccine development.

  5. Serosurvey for selected pathogens in free-ranging American black bears (Ursus americanus) in Maryland, USA.

    PubMed

    Bronson, Ellen; Spiker, Harry; Driscoll, Cindy P

    2014-10-01

    American black bears (Ursus americanus) in Maryland, USA, live in forested areas in close proximity to humans and their domestic pets. From 1999 to 2011, we collected 84 serum samples from 63 black bears (18 males; 45 females) in five Maryland counties and tested them for exposure to infectious, including zoonotic, pathogens. A large portion of the bears had antibody to canine distemper virus and Toxoplasma gondii, many at high titers. Prevalences of antibodies to zoonotic agents such as rabies virus and to infectious agents of carnivores including canine adenovirus and canine parvovirus were lower. Bears also had antibodies to vector-borne pathogens common to bears and humans such as West Nile virus, Borrelia burgdorferi, Rickettsia rickettsii, and Anaplasma phagocytophilum. Antibodies were detected to Leptospira interrogans serovars Pomona, Icterohaemorrhagiae, Canicola, Grippotyphosa, and Bratislava. We did not detect antibodies to Brucella canis or Ehrlichia canis. Although this population of Maryland black bears demonstrated exposure to multiple pathogens of concern for humans and domesticated animals, the low levels of clinical disease in this and other free-ranging black bear populations indicate the black bear is likely a spillover host for the majority of pathogens studied. Nevertheless, bear populations living at the human-domestic-wildlife interface with increasing human and domestic animal exposure should continue to be monitored because this population likely serves as a useful sentinel of ecosystem health.

  6. Sulfate uptake by crustacean hepatopancreatic brush border membrane vesicles. [Homarus americanus

    SciTech Connect

    Gerencser, G.A.; Cattey, M.A; Ahearn, G.A. Univ. of Hawaii, Honolulu )

    1990-02-26

    Purified brush border membrane vesicles (BBMV) were prepared from Atlantic lobster (Homarus americanus) hepatopancreas using differential centrifugation and Mg{sup +2} precipitation techniques. Uptake of 0.1 mM {sup 35}SO{sub 4}{sup {minus}2} was stimulated by pre-loading vesicles with Cl{sup {minus}} leading to a transient accumulation of isotope more than twice that at equilibrium. Pre-loading with HCO{sub 3}{sup {minus}} or gluconate had no effect on sulfate uptake. No stimulation of {sup 35}SO{sub 4}{sup {minus}2} was observed in the presence of inwardly directed Na{sup +} or tetramethylammonium{sup +} gradients. Uptake of the divalent anion was strongly stimulated by inwardly directed proton gradients (pH{sub o} < pH{sub i}) and markedly inhibited by outwardly directed proton gradients (pH{sub o} > pH{sub i}). {sup 35}SO{sub 4}{sup {minus}2}/Cl{sup {minus}} exchange was enhanced by imposing a transmembrane inside positive K{sup +} diffusion potential and inhibited by a membrane potential of the opposite polarity (K{sup +}/valinomycin). Results suggest the presence of a proton-dependent, electrogenic anion antiport mechanism in BBMV isolated from the crustacean hepatopancreas.

  7. Electrogenic sulfate uptake by crustacean hepatopancreatic basolateral membrane vesicles. [Homarus americanus

    SciTech Connect

    Cattey, M.A.; Gerencser, G.A.; Aheam, G.A. Univ. of Florida, Gainesville )

    1990-02-26

    Basolateral membrane vesicles (BLMV) were isolated from Atlantic lobster (Homarus americanus) hepatopancreas and purified by discontinuous sucrose gradient centrifugation. BLMV prepared in this fashion were osmotically reactive exhibiting linear dependence of vesicular {sup 35}SO{sub 4}{sup {minus}2} uptake to increasing external osmotic pressure with negligible non-specific isotope binding. Under short circuited conditions (valinomycin/K{sup +}) BLMV responded to either a HCO{sub 3}{sup {minus}} gradient directed out or equilibrated HCO{sub 3}{sup {minus}} (10 mM) by displaying short term accumulation of sulfate above that of equilibrium. Uptake of divalent anion was unaffected by an inwardly directed transmembrane Na{sup +} or tetramethylammonium{sup +} gradient. {sup 35}SO{sub 4}{sup {minus}2}/HCO{sub 3}{sup {minus}} exchange in the presence of valinomycin was stimulated by transient inside positive K{sup +} diffusion potentials and inhibited by transient inside negative K{sup +} diffusion potentials. The role of electrogenic anion exchange by hepatopancreas BLMV in transcellular sulfate transport is discussed.

  8. Temporal dynamics of genetic variability in a mountain goat (Oreamnos americanus) population.

    PubMed

    Ortego, Joaquín; Yannic, Glenn; Shafer, Aaron B A; Mainguy, Julien; Festa-Bianchet, Marco; Coltman, David W; Côté, Steeve D

    2011-04-01

    The association between population dynamics and genetic variability is of fundamental importance for both evolutionary and conservation biology. We combined long-term population monitoring and molecular genetic data from 123 offspring and their parents at 28 microsatellite loci to investigate changes in genetic diversity over 14 cohorts in a small and relatively isolated population of mountain goats (Oreamnos americanus) during a period of demographic increase. Offspring heterozygosity decreased while parental genetic similarity and inbreeding coefficients (F(IS) ) increased over the study period (1995-2008). Immigrants introduced three novel alleles into the population and matings between residents and immigrants produced more heterozygous offspring than local crosses, suggesting that immigration can increase population genetic variability. The population experienced genetic drift over the study period, reflected by a reduced allelic richness over time and an 'isolation-by-time' pattern of genetic structure. The temporal decline of individual genetic diversity despite increasing population size probably resulted from a combination of genetic drift due to small effective population size, inbreeding and insufficient counterbalancing by immigration. This study highlights the importance of long-term genetic monitoring to understand how demographic processes influence temporal changes of genetic diversity in long-lived organisms.

  9. Sarcocystis oreamni, n. sp. (Apicomplexa: Sarcocystidae) from the mountain goat (Oreamnos americanus).

    PubMed

    Calero-Bernal, Rafael; Van Wilpe, Erna; White, Kevin; Verma, Shiv K; Cerqueira-Cézar, Camila K; Dubey, Jitender P

    2015-11-01

    Numerous species of Sarcocystis have been reported from wild ruminants, but none has been named from the Rocky Mountain goat (Oreamnos americanus). Mature sarcocysts were found in frozen muscle samples of three of seven mountain goats from Alaska, USA. Two morphological types of sarcocysts were found; one had Sarcocystis cornagliai-like sarcocysts, previously named from the Alpine ibex (Capra ibex) from Europe. Two other goats were infected with a new species, Sarcocystis oreamni. Sarcocystis oreamni sarcocysts were microscopic with 2 μm-thick sarcocyst wall. By transmission electron microscopy, the sarcocyst wall had 1.7 μm-thick with unusual molar tooth-like villar protrusions (vp), type 29. The vp had an electron dense core and two disc-shaped plaques at the tip with fine microtubules. Bradyzoites were 8.6-9.1 μm long. Single nucleotide polymorphism (SNP) identified in 18S rRNA, and 28S rRNA loci of rDNA regions that suggested S. oreamni molecularly apart from related species. The phylogenetic analysis based on 18S rRNA and 28S rRNA sequences suggested S. oreamni is related with Sarcocystis species that employ members of the Canidae family as their definitive host.

  10. Bears "Count" Too: Quantity Estimation and Comparison in Black Bears (Ursus Americanus).

    PubMed

    Vonk, Jennifer; Beran, Michael J

    2012-07-01

    Studies of bear cognition are notably missing from the comparative record despite bears' large relative brain size and interesting status as generalist carnivores facing complex foraging challenges, but lacking complex social structures. We investigated the numerical abilities of three American black bears (Ursus Americanus) by presenting discrimination tasks on a touch-screen computer. One bear chose the larger of two arrays of dot stimuli, while two bears chose the smaller array of dots. On some trials the relative number of dots was congruent with the relative total area of the two arrays. On other trials number of dots was incongruent with area. All of the bears were above chance on trials of both types with static dots. Despite encountering greater difficulty with dots that moved within the arrays, one bear was able to discriminate numerically larger arrays of moving dots, and a subset of moving dots from within the larger array, even when area and number were incongruent. Thus, although the bears used area as a cue to guide responding, they were also able to use number as a cue. The pattern of performance was similar to that found previously with monkeys, and suggests that bears may also show other forms of sophisticated quantitative abilities.

  11. Social network analysis of mating patterns in American black bears (Ursus americanus).

    PubMed

    Moore, Jennifer A; Xu, Ran; Frank, Kenneth; Draheim, Hope; Scribner, Kim T

    2015-08-01

    Nonrandom mating can structure populations and has important implications for population-level processes. Investigating how and why mating deviates from random is important for understanding evolutionary processes as well as informing conservation and management. Prior to the implementation of parentage analyses, understanding mating patterns in solitary, elusive species like bears was virtually impossible. Here, we capitalize on a long-term genetic data set collected from black bears (Ursus americanus) (N = 2422) in the Northern Lower Peninsula (NLP) of Michigan, USA. We identified mated pairs using parentage analysis and applied logistic regression (selection) models that controlled for features of the social network, to quantify the effects of individual characteristics, and spatial and population demographic factors on mating dynamics. Logistic regression models revealed that black bear mating was associated with spatial proximity of mates, male age, the time a pair had coexisted, local population density and relatedness. Mated pairs were more likely to contain older males. On average, bears tended to mate with nearby individuals to whom they were related, which does not support the existence of kin recognition in black bears. Pairwise relatedness was especially high for mated pairs containing young males. Restricted dispersal and high male turnover from intensive harvest mortality of NLP black bears are probably the underlying factors associated with younger male bears mating more often with female relatives. Our findings illustrate how harvest has the potential to disrupt the social structure of game species, which warrants further attention for conservation and management. © 2015 John Wiley & Sons Ltd.

  12. Anaerobic oral flora in the North American black bear (Ursus americanus) in eastern North Carolina.

    PubMed

    Clarke, Elsburgh O; Stoskopf, Michael K; Minter, Larry J; Stringer, Elizabeth M

    2012-06-01

    Microbial flora can provide insight into the ecology and natural history of wildlife in addition to improving understanding of health risks. This study examines the anaerobic oral flora of hunter killed black bears (Ursus americanus) in eastern North Carolina. Oral swabs from the buccal and lingual supragingival tooth surfaces of the first and second mandibular and maxillary molars of 22 black bears were inoculated onto Brucella Blood Agar plates supplemented with hemin and vitamin K after transport from the field using reduced oxoid nutrient broth. Sixteen anaerobic bacterial species, representing nine genera were identified using the RapID ANA II Micromethod Kit system and a number of organisms grown that could not be identified with the system. The most frequently identified anaerobes were Peptostreptococcus prevotii, Streptococcus constellatus, and Porphyromonas gingivalis. The diversity in the anaerobic oral flora of black bear in eastern North Carolina suggests the importance of including these organisms in basic health risk assessment protocols and suggests a potential tool for assessment of bear/habitat interactions. Copyright © 2012 Elsevier Ltd. All rights reserved.

  13. Genomic analysis of expressed sequence tags in American black bear Ursus americanus.

    PubMed

    Zhao, Sen; Shao, Chunxuan; Goropashnaya, Anna V; Stewart, Nathan C; Xu, Yichi; Tøien, Øivind; Barnes, Brian M; Fedorov, Vadim B; Yan, Jun

    2010-03-26

    Species of the bear family (Ursidae) are important organisms for research in molecular evolution, comparative physiology and conservation biology, but relatively little genetic sequence information is available for this group. Here we report the development and analyses of the first large scale Expressed Sequence Tag (EST) resource for the American black bear (Ursus americanus). Comprehensive analyses of molecular functions, alternative splicing, and tissue-specific expression of 38,757 black bear EST sequences were conducted using the dog genome as a reference. We identified 18 genes, involved in functions such as lipid catabolism, cell cycle, and vesicle-mediated transport, that are showing rapid evolution in the bear lineage Three genes, Phospholamban (PLN), cysteine glycine-rich protein 3 (CSRP3) and Troponin I type 3 (TNNI3), are related to heart contraction, and defects in these genes in humans lead to heart disease. Two genes, biphenyl hydrolase-like (BPHL) and CSRP3, contain positively selected sites in bear. Global analysis of evolution rates of hibernation-related genes in bear showed that they are largely conserved and slowly evolving genes, rather than novel and fast-evolving genes. We provide a genomic resource for an important mammalian organism and our study sheds new light on the possible functions and evolution of bear genes.

  14. Right paw foraging bias in wild black bear (Ursus americanus kermodei).

    PubMed

    Reimchen, T E; Spoljaric, M A

    2011-07-01

    Using field observations of ~15 wild adult black bear (Ursus americanus kermodei) foraging on a salmon stream during two autumns on the central coast of British Columbia, we tested for laterality of forelimb use during lunging and during handling of salmon. Of 288 lunging events observed overall, 53% were non biased, 26% were right-limb biased, and 21% left-limb biased (p = .53 between left and right bias). Among six bears in which we could ascertain individual identity (182 lunging events), there was heterogeneity among individuals (p <.05) of which two were significantly right biased and one significantly left biased (p<.005). Of 186 carcass-handling (pick-up) events, 68% were right-pawed (p <.005) and there was no heterogeneity among five individually identifiable bears (p = .19). There was no forelimb laterality in adjustment of the prey in the mouth or in securing the prey to the substrate. This is the first report of task-specific behavioural lateralisation of a wild carnivore and is suggestive of a right bias (left-hemisphere dominance) in object manipulation.

  15. Liver lesions in winter flounder (Pseudopleuronectes americanus) from Jamaica Bay, New York: Indications of environmental degradation

    USGS Publications Warehouse

    Augspurger, T.P.; Herman, R.L.; Tanacredi, J.T.; Hatfield, J.S.

    1994-01-01

    Liver sections of winter flounder (Pseudopleuronectes americanus) collected from Jamaica Bay and Shinnecock Bay, New York, in 1989, were examined microscopically to determine the pervasiveness of liver lesions observed previously in Jamaica Bay winter flounder. Neoplastic lesions were not detected in fish from Jamaica Bay or the Shinnecock Bay reference site. Twenty-two percent of Jamaica Bay winter flounder examined (n = 103) had unusual vacuolization of hepatocytes and biliary pre-ductal and ductal cells (referred to hereafter as the vacuolated cell lesion). The lesion, identical to that found in 25% of Jamaica Bay winter flounder examined in 1988, has previously been identified in fishes taken from highly polluted regions of the Atlantic coast (e.g., Boston Harbor, Massachusetts, and Black Rock Harbor, Connecticut). Prevalence of the vacuolated cell lesion in winter flounder from Jamaica Bay was significantly greater (p < 0.0001) than in 102 specimens collected from Shinnecock Bay. Current scientific literature indicates vacuolated hepatocytes and cholangiocytes are chronically injured and that the extent of their deformity is consistent with the action of a hepatotoxicant. The high prevalence of vacuolated hepatocytes in Jamaica Bay winter flounder and absence of the lesion in flounder from reference sites strongly supports the hypothesis that this impairment is a manifestation of a toxic condition in at least some portions of Jamaica Bay.

  16. Seasonal variation in American black bear Ursus americanus activity patterns: Quantification via remote photography

    USGS Publications Warehouse

    Bridges, A.S.; Vaughan, M.R.; Klenzendorf, S.

    2004-01-01

    Activity pattern plasticity may serve as an evolutionary adaptation to optimize fitness in an inconstant environment, however, quantifying patterns and demonstrating variation can be problematic. For American black bears Ursus americanus, wariness and habitat inaccessibility further complicate quantification. Radio telemetry has been the primary technique used to examine activity, however, interpretation error and limitation on numbers of animals available to monitor prevent extrapolation to unmarked or untransmittered members of the population. We used remote cameras to quantify black bear activity patterns and examined differences by season, sex and reproductive class in the Alleghany Mountains of western Virginia, USA. We used 1,533 pictures of black bears taken during 1998-2002 for our analyses. Black bears generally were diurnal in summer and nocturnal in autumn with a vespertine activity peak during both seasons. Bear-hound training seasons occurred during September and may offer explanation for the observed shift towards nocturnal behaviour. We found no substantial differences in activity patterns between sex and reproductive classes. Use of remote cameras allowed us to efficiently sample larger numbers of individual animals and likely offered a better approximation of population-level activity patterns than individual-level, telemetry-based methodologies.

  17. Short- and long-term dietary effects on disease and mortality in American lobster Homarus americanus.

    PubMed

    Tlusty, Michael F; Myers, Anna; Metzler, Anita

    2008-01-24

    The American lobster Homarus americanus fishery is heavily dependent on the use of fish as bait to entice lobsters into traps. There is concern that this food supplementation is nutritionally insufficient for lobsters, but previous experiments reported conflicting results. We conducted a long-term feeding experiment in which 1 yr old American lobsters were fed one of 7 diets for a period of 352 d, a time that allowed the lobsters to molt thrice. The diets consisted of fresh frozen herring, a 'wild' diet (rock crab, mussel, and Spirulina algae), a formulated artificial diet for shrimp, paired combinations of these 3 diets or a diet formulated at the New England Aquarium (Artemia, fish and krill meal, Spirulina algae, soy lecithin, vitamins and minerals). The lobsters fed the diet of 100% fish had higher initial molting rates, but within the period of this experiment all either contracted shell disease or died. Mixed diets resulted in higher survival and a lower probability of mortality. This research demonstrated a critical time component to diet studies in lobsters. Short- and long-term impacts of diet differ. In the long term, continual high consumption rates of fish by the lobsters promote poor health in all lobsters, not just those of market size. The use of fish as bait may make lobsters more susceptible to the stress associated with environmental fluctuation, thereby leading to increased disease and mortality. This nutritional stress can be used to develop a laboratory model of shell disease in American lobsters.

  18. Behavioral response and kinetics of terrestrial atrazine exposure in American toads (bufo americanus)

    USGS Publications Warehouse

    Storrs, Mendez S.I.; Tillitt, D.E.; Rittenhouse, T.A.G.; Semlitsch, R.D.

    2009-01-01

    Amphibians in terrestrial environments obtain water through a highly vascularized pelvic patch of skin. Chemicals can also be exchanged across this patch. Atrazine (ATZ), a widespread herbicide, continues to be a concern among amphibian ecologists based on potential exposure and toxicity. Few studies have examined its impact on the terrestrial juvenile or adult stages of toads. In the current study, we asked the following questions: (1) Will juvenile American toads (Bufo americanus) avoid soils contaminated with ATZ? (2) Can they absorb ATZ across the pelvic patch? (3) If so, how is it distributed among the organs and eventually eliminated? We conducted a behavioral choice test between control soil and soil dosed with ecologically relevant concentrations of ATZ. In addition, we examined the uptake, distribution, and elimination of water dosed with 14C-labeled ATZ. Our data demonstrate that toads do not avoid ATZ-laden soils. ATZ crossed the pelvic patch rapidly and reached an apparent equilibrium within 5 h. The majority of the radiolabeled ATZ ended up in the intestines, whereas the greatest concentrations were observed in the gall bladder. Thus, exposure of adult life stages of amphibians through direct uptake of ATZ from soils and runoff water should be considered in risk evaluations. ?? 2009 Springer Science+Business Media, LLC.

  19. Bone formation is not impaired by hibernation (disuse) in black bears Ursus americanus

    USGS Publications Warehouse

    Donahue, S.W.; Vaughan, M.R.; Demers, L.M.; Donahue, H.J.

    2003-01-01

    Disuse by bed rest, limb immobilization or space flight causes rapid bone loss by arresting bone formation and accelerating bone resorption. This net bone loss increases the risk of fracture upon remobilization. Bone loss also occurs in hibernating ground squirrels, golden hamsters, and little brown bats by arresting bone formation and accelerating bone resorption. There is some histological evidence to suggest that black bears Ursus americanus do not lose bone mass during hibernation (i.e. disuse). There is also evidence suggesting that muscle mass and strength are preserved in black bears during hibernation. The question of whether bears can prevent bone loss during hibernation has not been conclusively answered. The goal of the current study was to further assess bone metabolism in hibernating black bears. Using the same serum markers of bone remodeling used to evaluate human patients with osteoporosis, we assayed serum from five black bears, collected every 10 days over a 196-day period, for bone resorption and formation markers. Here we show that bone resorption remains elevated over the entire hibernation period compared to the pre-hibernation period, but osteoblastic bone formation is not impaired by hibernation and is rapidly accelerated during remobilization following hibernation.

  20. Effects of cooking on levels of PCBs in the fillets of winter flounder (Pseudopleuronectes americanus)

    SciTech Connect

    Poston, T.M.; Durell, G.S.; Koczwara, G.; Spellacy, A.M.

    1995-08-01

    The Pacific Northwest Laboratory and Battelle Ocean Sciences performed a study to determine the effect of cooking on polychlorinated biphenyl (PCB) levels in the fillets of winter flounder (Pseudopleuronectes americanus). Broiling, pan frying, and deep frying in oil were tested on fillets from 21 fish collected from New Bedford Harbor, Massachusetts, on February 21, 1991. The evaluation involved estimating the change in PCB concentrations using a mass-balance approach that factored the change in fillet weight resulting from cooking with the changes in PCB concentration expressed on a precooked wet-weight basis. Deep frying in oil resulted in a 47% reduction in total PCB levels in fillet tissue. Additionally, deep frying caused a 40% reduction in fillet mass. Pan frying and broiling resulted in statistically in insignificant increases in total PCB levels of 15% and 17%, respectively. Fillet mass reductions resulting from pan frying and broiling were 7% and 15%, respectively. The effects of cooking on 18 individual congeners generally paralleled the results observed for total PCB. All 18 congeners were significantly reduced by deep frying. Congener Cl{sub 2}(08) also was significantly reduced by either pan frying. Congeners Cl{sub 5}(105) and Cl{sub 5}(118) showed apparent significant increases in concentrations following pan frying. Congeners Cl{sub 5}(105), Cl{sub 5}(118), and C1{sub 6}(138) showed significant increases in concentration following broiling.

  1. Experimental Infection and Detection of Necrotizing Hepatopancreatitis Bacterium in the American Lobster Homarus americanus

    PubMed Central

    Avila-Villa, Luz A.; Gollas-Galván, Teresa; Martínez-Porchas, Marcel; Mendoza-Cano, Fernando; Hernández-López, Jorge

    2012-01-01

    Necrotizing hepatopancreatitis bacterium (NHPB) is an obligated intracellular bacteria causing severe hepatopancreatic damages and mass mortalities in penaeid shrimp. The worldwide distribution of penaeid shrimp as alien species threatens the life cycle of other crustacean species. The aim of the experiment was to evaluate the possibility of experimentally infecting the American lobster (Homarus americanus) with NHPB extracted from shrimp hepatopancreas. Homogenates from infected shrimp were fed by force to lobsters. Other group of lobsters was fed with homogenates of NHPB-free hepatopancreas. After the 15th day from initial inoculation, the presence of NHPB was detected by polymerase chain reaction in feces and hepatopancreas from lobsters inoculated with infected homogenates. Necrotized spots were observed in the surface of lobster hepatopancreas. In contrast, lobsters fed on NHPB-free homogenates resulted negative for NHPB. Evidence suggests the plasticity of NHPB which can infect crustacean from different species and inhabiting diverse latitudes. Considering the results, the American lobster could be a good candidate to maintain available NHPB in vivo. PMID:22645497

  2. Effects of body temperature and hydration state on organismal performance of toads, Bufo americanus.

    PubMed

    Preest, Marion R; Pough, F Harvey

    2003-01-01

    Temperature and humidity are dominant environmental variables affecting performance of nocturnal, terrestrial amphibians. Toads are frequently active at body temperatures (T(b)) and hydration states (HS) that yield suboptimal performance. We investigated the combined effects of T(b) and HS on feeding, locomotion, and metabolism of Bufo americanus. More toads responded to the presence of prey when fully hydrated than when dehydrated, and times to orient to prey, maneuver around a barrier, and reach prey were less in hydrated than in dehydrated animals. Time to capture prey decreased with increasing T(b) in fully hydrated, but not dehydrated, toads, and hydrated animals caught prey more rapidly than did dehydrated animals. Distance traveled in 5 min and aerobic scope were affected by T(b). Generally, individuals that performed well in the feeding experiments at a particular T(b) and HS also performed well at a different T(b) and HS. The same was true for distance traveled and aerobic scope. However, within combinations of T(b) and HS, correlations between performance variables were minimal. Specialization of a particular variable resulting in high performance at a certain T(b) and HS does not appear to exact a cost in terms of performance at a different T(b) and HS.

  3. Energy expenditure during activity in the American lobster Homarus americanus: Correlations with body acceleration.

    PubMed

    Lyons, G N; Halsey, L G; Pope, E C; Eddington, J D; Houghton, J D R

    2013-10-01

    How animals manage time and expend energy has implications for survivorship. Being able to measure key metabolic costs of animals under natural conditions is therefore an important tool in behavioral ecology. One method for estimating activity-specific metabolic rate is via derived measures of acceleration, often 'overall dynamic body acceleration' (ODBA), recorded by an instrumented acceleration logger. ODBA has been shown to correlate well with rate of oxygen consumption (V˙o2) in a range of species during activity in the laboratory. This study devised a method for attaching acceleration loggers to decapod crustaceans and then correlated ODBA against concurrent respirometry readings to assess accelerometry as a proxy for activity-specific energy expenditure in a model species, the American lobster Homarus americanus. Where the instrumented animals exhibited a sufficient range of activity levels, positive linear relationships were found between V˙o2 and ODBA over 20min periods at a range of ambient temperatures (6, 13 and 20°C). Mixed effect linear models based on these data and morphometrics provided reasonably strong predictive power for estimating activity-specific V˙o2 from ODBA. These V˙o2-ODBA calibrations demonstrate the potential of accelerometry as an effective predictor of behavior-specific metabolic rate of crustaceans in the wild during periods of activity.

  4. Behavior and substrate selection during larval settling in the lobster Homarus americanus

    SciTech Connect

    Botero, L.; Atema, J.

    1982-02-01

    During the molt from third- to fourth-stage, larvae of Homarus americanus metamorphose into their adult form and pigmentation. In the course of the fourth stage, their pelagic life changes to a benthic existence. Artificial substrate choice experiments and qualitative illumination experiments show that during the early fourth stage the phototactic response reverses from positive to negative. Together with positive thigmotaxis, this results in a choice of dark crevices. Among natural substrate choices, preferential settlement occurred on macroalgal-covered rocks, followed by rocks on sand, mud, and sand. Although mud was not a preferred substrate in choice tests, the animals that chose mud and those that were presented only with mud settled successfully and were immediately efficient in their burrowing behavior, constructing U-shaped tunnels even in the center of the aquarium without using a pebble or rock as a starting point. These laboratory test confirm field observations that lobsters can successfully exploit a variety of substrates. They show that a substrate with preformed crevices is preferred for settling, but other substrates can be manipulated to make suitable burrows.

  5. Neuromuscular synapses on the dactyl opener muscle of the lobster Homarus americanus.

    PubMed

    Hamilton, Jonna; Dillaman, Richard M; Worden, Mary Kate

    2006-12-01

    The crustacean dactyl opener neuromuscular system has been studied extensively as a model system that exhibits several forms of synaptic plasticity. We report the ultrastructural features of the synapses on dactyl opener of the lobster (Homarus americanus) as determined by examination of serial thin sections. Several innervation sites supplied by an inhibitory motoneuron have been observed without nearby excitatory innervation, indicating that excitatory and inhibitory inputs to the muscle are not always closely matched. The ultrastructural features of the lobster synapses are generally similar to those described previously for the homologous crayfish muscle, with one major distinction: few dense bars are seen at the presynaptic membranes of these lobster synapses. The majority of the lobster neuromuscular synapses lack dense bars altogether, and the mean number of dense bars per synapse is relatively low. In view of the finding that the physiology of the lobster dactyl opener synapses is similar to that reported for crayfish, these ultrastructural observations suggest that the structural complexity of the synapses may not be a critical factor determining synaptic plasticity.

  6. Antimicrobial activity in the cuticle of the American lobster, Homarus americanus.

    PubMed

    Mars Brisbin, Margaret; McElroy, Anne E; Pales Espinosa, Emmanuelle; Allam, Bassem

    2015-06-01

    American lobster, Homarus americanus, continues to be an ecologically and socioeconomically important species despite a severe decline in catches from Southern New England and Long Island Sound (USA) and a high prevalence of epizootic shell disease in these populations. A better understanding of lobster immune defenses remains necessary. Cuticle material collected from Long Island Sound lobsters was found to be active against a broad spectrum of bacteria, including Gram-negative and -positive species. The antimicrobial activity was characterized by boiling, muffling, and size fractioning. Boiling did not significantly reduce activity, while muffling did have a significant effect, suggesting that the active component is organic and heat stable. Size fractioning with 3 and 10 kDa filters did not significantly affect activity. Fast protein liquid chromatography fractions were also tested for antimicrobial activity, and fractions exhibiting protein peaks remained active. MALDI mass spectrometry revealed peptide peaks at 1.6, 2.8, 4.6, and 5.6 kDa. The data presented suggest that one or several antimicrobial peptides contribute to antimicrobial activity present in the American lobster cuticle.

  7. An unusual cuticular tumor-like growth on the abdomen of a lobster, Homarus americanus.

    PubMed

    Shields, Jeffrey D; Small, Hamish J

    2013-11-01

    Tumors are rare in crustaceans, and whereas a few have been reported from the lobster Homarus americanus none have been adequately described. A lobster with an unusual, large, blue-colored tumor-like growth projecting laterally outward from the first abdominal somite was caught off Stonington, Maine, USA. The growth was rugose and covered by a relatively normal appearing cuticle with dispersed focal melanization. The underlying stroma consisted of an internal area of rescaffolded fibrous connective tissue, restructured muscle fibers, few arterioles, and an epidermal area comprised of columnar, highly vacuolated epithelial cells. No infectious pathogens or unusual inclusions were observed with microscopy and no eukaryotic pathogens were detected via molecular sequencing. Given the nature of the histology and the appearance of the growth, we identify the mass as a benign papilliform hamartoma that likely originated as a result of abnormal wound repair possibly initiated around ecdysis. This represents the first tumor-like hamartoma reported from a lobster, and the second hamartoma reported from a crustacean.

  8. Daily Rhythms of PERIOD protein in the eyestalk of the American lobster, Homarus americanus.

    PubMed

    Grabek, Katharine R; Chabot, Christopher C

    2012-01-01

    The daily rhythm of PERIOD protein (PER) expression is an integral component of the circadian clock, which is found among a broad range of animal species including fruit flies, marine mollusks and even humans. The use of antibodies directed against PER has provided a helpful tool in the discovery of PER homologues and the labeling of putative pacemaker cells, especially in animals for which an annotated genome is not readily available. In this study, DrosophilaPER antibodies were used to probe for PER in the American lobster, Homarus americanus. This species exhibits robust endogenous circadian rhythms but the circadian clock has yet to be located or characterized. PER was detected in the eyestalks of the lobster but not in the brain. Furthermore, a significant effect of the LD cycle on daily PER abundance was identified, and PER was significantly more abundant at mid dark than in early light or mid light hours. Our results suggest that PER is a part of the molecular machinery of the circadian clock located in the eyestalk of the lobster.

  9. American Lobsters (Homarus Americanus) not Surviving During Air Transport: Evaluation of Microbial Spoilage

    PubMed Central

    Tirloni, Erica; Stella, Simone; Gennari, Mario; Colombo, Fabio

    2016-01-01

    Eighteen American lobsters (Homarus americanus), dead during air transport, were analysed in order to evaluate the microbial population of meat, gills and gut: no specific studies have ever been conducted so far on the microbiological quality of American lobsters’ meats in terms of spoilage microbiota. The meat samples showed very limited total viable counts, in almost all the cases below the level of 6 Log CFU/g, while higher loads were found, as expected, in gut and gills, the most probable source of contamination. These data could justify the possibility to commercialise these not-surviving subjects, without quality concerns for the consumers. Most of the isolates resulted to be clustered with type strains of Pseudoalteromonas spp. (43.1%) and Photobacterium spp. (24.1%), and in particular to species related to the natural marine environment. The distribution of the genera showed a marked inhomogeneity among the samples. The majority of the isolates identified resulted to possess proteolytic (69.3%) and lipolytic ability (75.5%), suggesting their potential spoilage ability. The maintanance of good hygienical practices, especially during the production of ready-to-eat lobsters-based products, and a proper storage could limit the possible replication of these microorganisms. PMID:27800442

  10. Genomic analysis of expressed sequence tags in American black bear Ursus americanus

    PubMed Central

    2010-01-01

    Background Species of the bear family (Ursidae) are important organisms for research in molecular evolution, comparative physiology and conservation biology, but relatively little genetic sequence information is available for this group. Here we report the development and analyses of the first large scale Expressed Sequence Tag (EST) resource for the American black bear (Ursus americanus). Results Comprehensive analyses of molecular functions, alternative splicing, and tissue-specific expression of 38,757 black bear EST sequences were conducted using the dog genome as a reference. We identified 18 genes, involved in functions such as lipid catabolism, cell cycle, and vesicle-mediated transport, that are showing rapid evolution in the bear lineage Three genes, Phospholamban (PLN), cysteine glycine-rich protein 3 (CSRP3) and Troponin I type 3 (TNNI3), are related to heart contraction, and defects in these genes in humans lead to heart disease. Two genes, biphenyl hydrolase-like (BPHL) and CSRP3, contain positively selected sites in bear. Global analysis of evolution rates of hibernation-related genes in bear showed that they are largely conserved and slowly evolving genes, rather than novel and fast-evolving genes. Conclusion We provide a genomic resource for an important mammalian organism and our study sheds new light on the possible functions and evolution of bear genes. PMID:20338065

  11. Immobilization of black bears (Ursus americanus) with orally administered carfentanil citrate.

    PubMed

    Ramsay, E C; Sleeman, J M; Clyde, V L

    1995-07-01

    Ten black bears (Ursus americanus) were immobilized with orally administered carfentanil citrate. The total carfentanil dose was mixed with 5 to 20 ml honey and given incrementally to captive bears. The bears ranged in weight from 80 (estimated) to 233 kg. Total carfentanil doses ranged from 0.7 to 3.0 mg, resulting in dosages of 6.8 to 18.8 micrograms carfentanil/kg. Mean (+/- SD) times from estimated 80% mixture consumption to sternal recumbency, and first safe human contact were 7.7 +/- 2.3 min and 19.7 +/- 5.6 min, respectively. Undesired side effects of immobilization were muscle rigidity, bradypnea, and oxygen desaturation. All bears received diazepam to alleviate muscle rigidity and were insufflated with oxygen during immobilization. Nine immobilizations were considered satisfactory or good. The bear receiving 6.8 micrograms carfentanil/kg, the lowest dosage used, was very excited during induction and required intravenous (IV) ketamine to permit safe examination. Immobilization was reversed with 100 mg naltrexone/mg carfentanil administered (75% subcutaneous, 25% IV). Bears recovered to full mobility in 6.3 +/- 1.9 min. Five bears vomited post-recovery but no episodes of renarcotization were observed.

  12. Modulation of gene expression in heart and liver of hibernating black bears (Ursus americanus).

    PubMed

    Fedorov, Vadim B; Goropashnaya, Anna V; Tøien, Oivind; Stewart, Nathan C; Chang, Celia; Wang, Haifang; Yan, Jun; Showe, Louise C; Showe, Michael K; Barnes, Brian M

    2011-03-31

    Hibernation is an adaptive strategy to survive in highly seasonal or unpredictable environments. The molecular and genetic basis of hibernation physiology in mammals has only recently been studied using large scale genomic approaches. We analyzed gene expression in the American black bear, Ursus americanus, using a custom 12,800 cDNA probe microarray to detect differences in expression that occur in heart and liver during winter hibernation in comparison to summer active animals. We identified 245 genes in heart and 319 genes in liver that were differentially expressed between winter and summer. The expression of 24 genes was significantly elevated during hibernation in both heart and liver. These genes are mostly involved in lipid catabolism and protein biosynthesis and include RNA binding protein motif 3 (Rbm3), which enhances protein synthesis at mildly hypothermic temperatures. Elevated expression of protein biosynthesis genes suggests induction of translation that may be related to adaptive mechanisms reducing cardiac and muscle atrophies over extended periods of low metabolism and immobility during hibernation in bears. Coordinated reduction of transcription of genes involved in amino acid catabolism suggests redirection of amino acids from catabolic pathways to protein biosynthesis. We identify common for black bears and small mammalian hibernators transcriptional changes in the liver that include induction of genes responsible for fatty acid β oxidation and carbohydrate synthesis and depression of genes involved in lipid biosynthesis, carbohydrate catabolism, cellular respiration and detoxification pathways. Our findings show that modulation of gene expression during winter hibernation represents molecular mechanism of adaptation to extreme environments.

  13. A descriptive analysis of the climbing mechanics of a mountain goat (Oreamnos americanus).

    PubMed

    Lewinson, Ryan T; Stefanyshyn, Darren J

    2016-12-01

    The mountain goat (Oreamnos americanus) is one of the most extraordinary mountaineers in the animal kingdom. While observational descriptions exist to indicate factors that may influence their climbing ability, these have never been assessed biomechanically. Here, we describe whole-body motion of a mountain goat during ascent of a 45° incline based on a video recording in the Canadian Rocky Mountains, and discuss the results in a mechanical context. During the push-off phase, the hindlimb extended and the forelimb was tucked close to the torso. During the pull-up phase, the hindlimb was raised near to the torso, while the forelimb humerus seemed to "lock" in a constant position relative to the torso, allowing the elbow to be held in close proximity to the whole-body center of mass. Extension of the elbow and carpal joints resulted in a vertical translation of the center of mass up the mountain slope. Based on the observations from this naturalistic study, hypotheses for future controlled studies of mountain goat climbing mechanics are proposed. Copyright © 2016 Elsevier GmbH. All rights reserved.

  14. Idiopathic lesions and visual deficits in the american lobster (Homarus americanus) from Long Island Sound, NY.

    PubMed

    Magel, Christopher R; Shields, Jeffrey D; Brill, Richard W

    2009-08-01

    In 1999, a mass mortality of the American lobster (Homarus americanus) occurred in western Long Island Sound (WLIS). Although the etiology of this event remains unknown, bottom water temperature, hypoxia, heavy metal poisoning, and pesticides are potential causal factors. Lobsters from WLIS continue to display signs of morbidity, including lethargy and cloudy grey eyes that contain idiopathic lesions. As the effect of these lesions on lobster vision is unknown, we used electroretinography (ERG) to document changes in visual function in lobsters from WLIS, while using histology to quantify the extent of physical damage. Seventy-three percent of lobsters from WLIS showed damage to photoreceptors and optic nerve fibers, including necrosis, cellular breakdown, and hemocyte infiltration in the optic nerves, rhabdoms, and ommatidia. Animals with more than 15% of their photoreceptors exhibiting damage also displayed markedly reduced responses to 10-ms flashes of a broad-spectrum white light. Specifically, maximum voltage (Vmax) responses were significantly lower and occurred at a lower light intensity compared to responses from lobsters lacking idiopathic lesions. Nearly a decade after the 1999 mortality event, lobsters from WLIS still appear to be subjected to a stressor of unknown etiology that causes significant functional damage to the eyes.

  15. Excretory calcinosis: a new fatal disease of wild American lobsters Homarus americanus.

    PubMed

    Dove, Alistair D M; LoBue, Carl; Bowser, Paul; Powell, Mark

    2004-03-10

    A significant number of moribund and dead lobsters Homarus americanus were reported to New York state authorities by lobster fishers in Long Island Sound (LIS) during the summer of 2002. Morbid lobsters were characterised by an orange discolouration of the abdomen, lethargy, an excess of epibionts and poor post-capture survival. On necropsy, severe extensive multifocal or diffuse mineralised granulomatous inflammation of the gills and antennal glands was the most striking pathology. In the gills, granulomas often occluded the lumen of filaments, resulting in congestion, ischemia and coagulative necrosis of gill tissues. In the antennal glands, granulomas were concentrated along the border between the coelomosac and labyrinth. No significant pathogens were recovered from diseased individuals. In prechronic individuals, however, it was evident that granulomas were focused around calcium carbonate (aragonite) crystals. This disease may result from anomalously high sea-bottom temperatures in LIS (approximately 23 degrees C) during the summer of 2002 and associated disruptions of the calcium chemistry of lobsters in favour of deposition of minerals in soft tissues. The ultimate cause of death of affected lobsters is probably respiratory failure due to reduced effective surface area of the gills, exacerbated by hypermetabolic temperatures and an abundance of epibionts.

  16. Histopathology of idiopathic lesions in the eyes of Homarus americanus from Long Island Sound.

    PubMed

    Maniscalco, Andrea M; Shields, Jeffrey D

    2006-02-01

    In 1999, American lobsters, Homarus americanus, from western Long Island Sound (WLIS) experienced a significant mortality. In 2001 and 2004, the eyes and eyestalks of lobsters from WLIS and central LIS were examined for histopathological changes. Idiopathic lesions were identified in the ommatidia and optic nerve fibers proximal to the ommatidia in 29 (56%) of the lobsters from LIS. Lesions were categorized as either moderate or severe. Moderate lesions had altered rhabdoms, clumped pigment, and altered optic nerve fibers. Severe lesions were marked by absent rhabdoms, clumped pigment in both the ommatidial region and in the optic nerve region; and optic nerve fibers that had been completely destroyed and were replaced by vascular tissue. Idiopathic lesions occurred primarily in the central and ventral regions of the eye, and with much less frequency in the dorsal region. In addition, damage to the dorsal area tended to occur only when the severity of lesions was high, indicating a spatially progressive pattern to the lesion development. The lesions occurred in both western and central Long Island Sound, with no significant differences in severity between locations. The prevalence of lesions did not vary between years, but in 2004, several eyes had less severe pathology than those from 2001. These data indicate that the etiological agent is present throughout a large portion of the Sound, and that lobsters are probably continually exposed to it.

  17. Microfibrils provide non-linear elastic behaviour in the abdominal artery of the lobster Homarus americanus.

    PubMed Central

    McConnell, C J; DeMont, M E; Wright, G M

    1997-01-01

    1. Microfibrils are becoming increasingly recognized as an important component of the extra-cellular matrix. However, almost nothing is known about their mechanical role in the diversity of tissues in which they are found. 2. Microfibrils form the principal structural component in the wall of the abdominal artery of the lobster Homarus americanus. We have used previous estimates of the mechanical properties of these microfibrils, estimates of the fraction of the aorta wall volume occupied by the microfibrils, and their angular distribution as a function of strain in a numerical model that predicts the macroscopic mechanical properties of the whole tissue. 3. Microfibrils alone, when their reorientation and deformation are accounted for, characterize the stress-strain behaviour of the vessel. Evidence of the evolutionary conservation of fibrillin between medusans, echinoderms and vertebrates implies that the mechanical properties of lobster microfibrils may apply to microfibrillar function in other taxa. This will have profound implications on the perceived roles of microfibrils in development, physiology and disease. Images Figure 2 Figure 3 Figure 4 PMID:9080378

  18. Dietary lipid requirements of adult lobsters, Homarus americanus (M.E.).

    PubMed

    Castell, J D; Covey, J F

    1976-08-01

    The effects of several dietary lipids on adult American lobster (Homarus americanus) were assessed over a 10 month feeding period. Cod liver oil (CLO) resulted in greater precent weight gains, feed conversion, percent edible meat and higher serum protein and hemocyte counts than either corn oil (CO) or hydrogenated coconut oil (HCO). These differences were probably due to an essential fatty acid (EFA) requirement by the lobster for linolenic series omega3 or other fatty acids present in CLO. The linoleic or omega6 fatty acids of CO appeared to have some sparing effect on several of the EFA deficiency symptoms. It was found that 5% CLO was optimal for mean percent weight gain, molt incidence, feed conversion and hemocyte counts of lobsters. Further increases in dietary CLO to 10% and 15% resulted in no significant improvement of any of the condition indices used. There was a decrease in serum protein and calcium when lobsters were fed a non-saponifiable sterol deficient diet. The addition of 1% cholesterol to the diet raised the serum protein, but resulted in even a greater decrease in the serum calcium level.

  19. Influenza virus antigenicity and broadly neutralizing epitopes.

    PubMed

    Air, Gillian M

    2015-04-01

    A vaccine formulation that would be effective against all strains of influenza virus has long been a goal of vaccine developers, but antibodies after infection or vaccination were seen to be strain specific and there was little evidence of cross-reactive antibodies that neutralized across subtypes. Recently a number of broadly neutralizing monoclonal antibodies have been characterized. This review describes the different classes of broadly neutralizing antibodies and discusses the potential of their therapeutic use or for design of immunogens that induce a high proportion of broadly neutralizing antibodies.

  20. Differential expression of American lobster (Homarus americanus) immune related genes during infection of Aerococcus viridans var. homari, the causative agent of Gaffkemia.

    PubMed

    Clark, K Fraser; Acorn, Adam R; Greenwood, Spencer J

    2013-02-01

    This is the first transcriptomic study focusing on immunity in the commercially valuable American lobster (Homarus americanus). We have conducted an in vivo infection trial using the Gram-positive bacterium Aerococcus viridans var. homari to determine how H. americanus responds to this naturally occurring lethal-pathogen. A novel H. americanus microarray was used to measure the transcriptomic changes occurring in over 14,000 genes in the lobster hepatopancreas. Hundreds of new immune genes and isoforms were identified and measured for the first time in this species, and our findings highlight 148 genes of interest involved in H. americanus pathogen response. We verified our microarray results using RT-qPCR on three anti-lipopolysaccharide (ALFHa-1, ALFHa-2, ALFHa-4), a thioredoxin, acute phase serum amyloid protein A, hexokinase and two trypsin genes. RT-qPCR and microarray findings show close agreement and highlight the significant increase in gene expression in many lobster immune genes during A. viridans infection. Differential expression of the ALFHa isoforms may indicate that the H. americanus immune response can be tailored to the class of pathogen causing disease.

  1. Seasonal variability of prey in the stomachs of Astropecten americanus (Echinodermata: Asteroidea) from off southern New England, U.S.A.

    NASA Astrophysics Data System (ADS)

    Franz, David R.; Worley, E. K.

    1982-04-01

    The diet of Astropecten americanus from a location off Nantucket Shoals, Massachusetts, was determined by analysis of stomach contents in May and November. Molluscs, followed by crustaceans, dominated stomach contents. The proportion of molluscan species in the diet differed significantly from the benthos, Juvenile Arctica islandica dominated stomachs in May, but declined sharply in November, probably due in part to predation by Astropecten. Other molluscs, not prevalent in the benthos, were important in stomach contents. Frequency distributions of counts of prey species in a sample of seastar stomachs indicate that some seastars ingest unusually large numbers of certain prey species. A. americanus generally ingests small prey individuals (<4mm) although larger prey are consumed, especially in November. Relatively high selectivity, seasonal shifts in the diversity of stomach contents, and evidence for patch-specific foraging are characteristics of A. americanus which are consistent with general predictions of optimum diet models.

  2. Neutralization of both IL-1α/IL-1β plays a major role in suppressing combined cigarette smoke/virus-induced pulmonary inflammation in mice.

    PubMed

    Bucher, Hannes; Mang, Samuel; Keck, Martina; Przibilla, Michèl; Lamb, David; Schiele, Felix; Wittenbrink, Mareike; Fuchs, Klaus; Jung, Birgit; Erb, Klaus J; Peter, Daniel

    2017-03-15

    Smoking is an important risk factor for the development of chronic obstructive pulmonary disease (COPD) and viral infections are believed to be major triggers of exacerbations, which periodically lead to a worsening of symptoms. The pro-inflammatory IL-1 family members IL-1α and IL-1β are increased in COPD patients and might contribute to disease pathology. We investigated whether individual or combined inhibition of these cytokines reduced lung inflammation in cigarette smoke (CS)-exposed and H1N1-infected BALB/c mice. Animals were treated with individual or combined antibodies (Abs) directed against IL-1α, IL-1β or IL-1R1. Cells in BAL fluid and cytokines/chemokines in lung homogenate were determined. The viral load was investigated. Blocking IL-1α had significant suppressive effects on total cells, neutrophils, and macrophages. Furthermore, it reduced KC levels significantly. Blocking of IL-1β did not provide significant activity. In line with the in vivo findings, IL-1α Abs but not IL-1β Abs reduced levels of TNF-α and IL-6 in H1N1 infected primary human bronchial epithelial air-liquid-interface cell culture. Concomitant usage of Abs against IL-1α/IL-1β revealed strong effects in vivo and reduced total cells, neutrophils and macrophages. Additionally, levels of KC, IL-6, TNF-α, MCP-1, MIP-1α and MIP-1β were significantly reduced and ICAM-1 and MUC5 A/C mRNA expression was attenuated. The viral load decreased significantly upon combined IL-1α/IL-1β Ab treatment. Blocking the IL-1R1 provided significant effects on total cells, neutrophils and macrophages but was inferior compared to inhibiting both its soluble ligands IL-1α/IL-1β. Our results suggest that combined inhibition of IL-1α/IL-1β might be beneficial to reduce CS/H1N1-induced airway inflammation. Moreover, combined targeting of both IL-1α/IL-1β might be more efficient compared to individual neutralization IL-1α or IL-1β or inhibition of the IL-1R1.

  3. Measurement of the total flux averaged neutrino induced neutral current elastic scattering cross section with the T2K Pi-Zero detector

    NASA Astrophysics Data System (ADS)

    Ruterbories, Daniel

    Tokai-to-Kamioka (T2K) is a second generation accelerator neutrino oscillation experiment. T2K uses a high intensity proton beam produced at the Japan Proton Accelerator Research Complex (J-PARC) incident on a carbon target and focused with three magnetic horns to produce a high intensity and nearly pure muon neutrino beam with a peak energy of 600 MeV at a 2.5º axis angle. The muon neutrino beam travels 295 km across Japan to the Super Kamiokande (SK) water Cherenkov detector in the Kamioka mine. The neutrino beam is also sampled by a complex of near detectors 280 m downstream of the carbon target located both on and off the beam axis. These detectors measure the neutrino beam before neutrino oscillations occur to provide input constraints to oscillation searches using SK. The off-axis near detector, ND280, is a composite detector made up of a tracker section and a Pi-Zero detector (POD), all surrounded by an electromagnetic calorimeter. The entire detector is enclosed in a dipole magnet with a field of 0.2 T. The primary purpose of the tracker section is to measure neutrino induced charged current events characterized by the production of muons. The POD is primarily designed to detect electromagnetic showers and to measure interactions on water through the use of a removable water target. In addition to these measurements, the ND280 detector is also used to study the cross sections of neutrino interactions on the various materials in the detectors. Limited knowledge of the cross sections in this neutrino energy regime are an important source of systematic error in neutrino oscillation measurements. This thesis presents a measurement of one neutrino interaction channel in the POD, neutral current elastic scattering (NCE). In this process a neutrino elastically scatters off a proton or neutron in the target nucleus producing a proton or neutron with higher energy. The signature of this process is a single proton track. A particle identification algorithm (PID) was

  4. Necator americanus and helminth co-infections: further down-modulation of hookworm-specific type 1 immune responses.

    PubMed

    Geiger, Stefan Michael; Alexander, Neal Douglas Edward; Fujiwara, Ricardo Toshio; Brooker, Simon; Cundill, Bonnie; Diemert, David Joseph; Correa-Oliveira, Rodrigo; Bethony, Jeffrey Michael

    2011-09-01

    Helminth co-infection in humans is common in tropical regions of the world where transmission of soil-transmitted helminths such as Ascaris lumbricoides, Trichuris trichiura, and the hookworms Necator americanus and Ancylostoma duodenale as well as other helminths such as Schistosoma mansoni often occur simultaneously. We investigated whether co-infection with another helminth(s) altered the human immune response to crude antigen extracts from either different stages of N. americanus infection (infective third stage or adult) or different crude antigen extract preparations (adult somatic and adult excretory/secretory). Using these antigens, we compared the cellular and humoral immune responses of individuals mono-infected with hookworm (N. americanus) and individuals co-infected with hookworm and other helminth infections, namely co-infection with either A. lumbricoides, Schistosoma mansoni, or both. Immunological variables were compared between hookworm infection group (mono- versus co-infected) by bootstrap, and principal component analysis (PCA) was used as a data reduction method. Contrary to several animal studies of helminth co-infection, we found that co-infected individuals had a further downmodulated Th1 cytokine response (e.g., reduced INF-γ), accompanied by a significant increase in the hookworm-specific humoral immune response (e.g. higher levels of IgE or IgG4 to crude antigen extracts) compared with mono- infected individuals. Neither of these changes was associated with a reduction of hookworm infection intensity in helminth co-infected individuals. From the standpoint of hookworm vaccine development, these results are relevant; i.e., the specific immune response to hookworm vaccine antigens might be altered by infection with another helminth.

  5. Relationship between body size and habitat complexity preference in age-0 and -1 year winter flounder Pseudopleuronectes americanus.

    PubMed

    Pappal, A L; Rountree, R A; MacDonald, D G

    2012-07-01

    The interaction between body size, habitat complexity and interstice width on habitat preference of age-0 and -1 year Pseudopleuronectes americanus was examined using continuous remote video observation. The habitat choices of juvenile P. americanus were recorded over a 6 h period in tanks with four treatments: bare sand, sand with low complexity cobble, sand with intermediate complexity cobble and sand with high complexity cobble. Both age-0 and -1 year fish preferred cobble to bare sand. Within cobble treatments, age-0 year fish preferred intermediate complexity cobble, with a 1.59 ratio of interstitial space to body width. The largest age-1 year fish (123-130 mm standard length, L(S) ) preferred low complexity cobble. While a significant preference was not detected, medium age-1 year fish (83-88 mm L(S) ) tended to select low complexity cobble, whereas small age-1 year fish (73-82 mm L(S) ) tended to select low and intermediate cobble, with an interstitial space to body width ratio of 1.05. For medium and large age-1 year fish, there was an increased selection of low complexity cobble, corresponding to larger interstitial space to body size ratios. This study indicates that juvenile P. americanus prefer complex habitat to unstructured habitat and that this preference is mediated by a relationship between fish body size and the size of structure interstices. These results contribute to the growing body of knowledge of complex habitat selection and drivers of habitat choice in flatfishes. © 2012 The Authors. Journal of Fish Biology © 2012 The Fisheries Society of the British Isles.

  6. De novo transcriptome assembly for the lobster Homarus americanus and characterization of differential gene expression across nervous system tissues.

    PubMed

    McGrath, Lara Lewis; Vollmer, Steven V; Kaluziak, Stefan T; Ayers, Joseph

    2016-01-16

    The American lobster, Homarus americanus, is an important species as an economically valuable fishery, a key member in marine ecosystems, and a well-studied model for central pattern generation, the neural networks that control rhythmic motor patterns. Despite multi-faceted scientific interest in this species, currently our genetic resources for the lobster are limited. In this study, we de novo assemble a transcriptome for Homarus americanus using central nervous system (CNS), muscle, and hybrid neurosecretory tissues and compare gene expression across these tissue types. In particular, we focus our analysis on genes relevant to central pattern generation and the identity of the neurons in a neural network, which is defined by combinations of genes distinguishing the neuronal behavior and phenotype, including ion channels, neurotransmitters, neuromodulators, receptors, transcription factors, and other gene products. Using samples from the central nervous system (brain, abdominal ganglia), abdominal muscle, and heart (cardiac ganglia, pericardial organs, muscle), we used RNA-Seq to characterize gene expression patterns across tissues types. We also compared control tissues with those challenged with the neuropeptide proctolin in vivo. Our transcriptome generated 34,813 transcripts with known protein annotations. Of these, 5,000-10,000 of annotated transcripts were significantly differentially expressed (DE) across tissue types. We found 421 transcripts for ion channels and identified receptors and/or proteins for over 20 different neurotransmitters and neuromodulators. Results indicated tissue-specific expression of select neuromodulator (allostatin, myomodulin, octopamine, nitric oxide) and neurotransmitter (glutamate, acetylcholine) pathways. We also identify differential expression of ion channel families, including kainite family glutamate receptors, inward-rectifying K(+) (IRK) channels, and transient receptor potential (TRP) A family channels, across central

  7. Fatal Caprine arthritis encephalitis virus-like infection in 4 Rocky Mountain goats (Oreamnos americanus).

    PubMed

    Patton, Kristin M; Bildfell, Robert J; Anderson, Mark L; Cebra, Christopher K; Valentine, Beth A

    2012-03-01

    Over a 3.5-year period, 4 Rocky Mountain goats (Oreamnos americanus), housed at a single facility, developed clinical disease attributed to infection by Caprine arthritis encephalitis virus (CAEV). Ages ranged from 1 to 10 years. Three of the goats, a 1-year-old female, a 2-year-old male, and a 5-year-old male, had been fed raw domestic goat milk from a single source that was later found to have CAEV on the premises. The fourth animal, a 10-year-old male, had not ingested domestic goat milk but had been housed with the other 3 Rocky Mountain goats. All 4 animals had clinical signs of pneumonia prior to death. At necropsy, findings in lungs included marked diffuse interstitial pneumonia characterized histologically by severe lymphoplasmacytic infiltrates with massive alveolar proteinosis, interstitial fibrosis, and type II pneumocyte hyperplasia. One animal also developed left-sided hemiparesis, and locally extensive lymphoplasmacytic myeloencephalitis was present in the cranial cervical spinal cord. Two animals had joint effusions, as well as severe lymphoplasmacytic and ulcerative synovitis. Immunohistochemical staining of fixed sections of lung tissue from all 4 goats, as well as spinal cord in 1 affected animal, and synovium from 2 affected animals were positive for CAEV antigen. Serology testing for anti-CAEV antibodies was positive in the 2 goats tested. The cases suggest that Rocky Mountain goats are susceptible to naturally occurring CAEV infection, that CAEV from domestic goats can be transmitted to this species through infected milk and by horizontal transmission, and that viral infection can result in clinically severe multisystemic disease.

  8. Multilocus heterozygosity, parental relatedness and individual fitness components in a wild mountain goat, Oreamnos americanus population.

    PubMed

    Mainguy, Julien; Côté, Steeve D; Coltman, David W

    2009-05-01

    Matings between relatives lead to a decrease in offspring genetic diversity which can reduce fitness, a phenomenon known as inbreeding depression. Because alpine ungulates generally live in small structured populations and often exhibit a polygynous mating system, they are susceptible to inbreeding. Here, we used marker-based measures of pairwise genetic relatedness and inbreeding to investigate the fitness consequences of matings between relatives in a long-term study population of mountain goats (Oreamnos americanus) at Caw Ridge, Alberta, Canada. We first assessed whether individuals avoided mating with kin by comparing actual and random mating pairs according to their estimated genetic relatedness, which was derived from 25 unlinked polymorphic microsatellite markers and reflected pedigree relatedness. We then examined whether individual multilocus heterozygosity H, used as a measure of inbreeding, was predicted by parental relatedness and associated with yearling survival and the annual probability of giving birth to a kid in adult females. Breeding pairs identified by genetic parentage analyses of offspring that survived to 1 year of age were less genetically related than expected under random matings. Parental relatedness was negatively correlated with offspring H, and more heterozygous yearlings had higher survival to 2 years of age. The probability of giving birth was not affected by H in adult females. Because kids that survived to yearling age were mainly produced by less genetically related parents, our results suggest that some individuals experienced inbreeding depression in early life. Future research will be required to quantify the levels of gene flow between different herds, and evaluate their effects on population genetic diversity and dynamics.

  9. Sequential ovulation and fertility of polyoestrus in American black bears (Ursus americanus).

    PubMed

    Himelright, Brendan M; Moore, Jenna M; Gonzales, Ramona L; Mendoza, Alejandra V; Dye, Penny S; Schuett, Randall J; Durrant, Barbara S; Read, Betsy A; Spady, Thomas J

    2014-01-01

    American black bears (Ursus americanus) are seasonally polyoestrous and exhibit delayed implantation, which may allow equal and independent fertility of recurrent oestruses of a mating season. We postulated that the luteal inactivity during delayed implantation allows bears to have sequential ovulation during a polyoestrous mating season such that each oestrus of a polyoestrous female will have equivalent fertility, and pregnancy would not preclude subsequent ovulation and superfetation. Controlled mating experiments were conducted on semi-free-ranging female American black bears during three mating seasons, wherein females were bred by different male cohorts in each oestrus. Behavioural observation, vulva score ranking, genetic paternity analysis, gross morphology of ovaries and microscopic morphology of diapaused embryos were used to evaluate the fertility of each subsequent oestrus in polyoestrous females. Oestrus duration, number of successful mounts and median vulva scores were similar between first and subsequent oestruses of the season. Polyoestrus occurred in 81.3% of oestrous females, with a 9.7 ± 5.5 day (mean ± SD) inter-oestrous interval. Sequential ovulation was documented in three polyoestrous females, including one that possessed both a corpus haemorrhagicum and a developed corpus luteum. Among polyoestrous dams, four of nine embryos were conceived in the first oestrus and five of nine in the second oestrus. These results indicate that each oestrus of polyoestrous females is capable of fertility, even if the female is already pregnant from a prior oestrus. Although superfetation was not directly observed in the present study, our results strongly suggest the potential of superfetation in the American black bear and provide novel insight into the complex behavioural and physiological breeding mechanisms of bears. Given that most endangered bear species share similar reproductive traits with American black bears, captive breeding programmes

  10. Prevalence of Babesia spp., Ehrlichia spp., and Tick Infestations in Oklahoma Black Bears (Ursus americanus).

    PubMed

    Skinner, Delaina; Mitcham, Jessica R; Starkey, Lindsay A; Noden, Bruce H; Fairbanks, W Sue; Little, Susan E

    2017-06-28

    American black bears (Ursus americanus) are commonly infested with ticks throughout their range, but there are few surveys for tick-borne disease agents in bears. To characterize tick infestations and determine the prevalence of current infection with Babesia spp. and past or current infection with Ehrlichia spp. in newly re-established populations of black bears in east central and southeastern Oklahoma, we identified adult (n=1,048) and immature (n=107) ticks recovered from bears (n=62). We evaluated serum and whole blood samples from a subset (n=49) for antibodies reactive to, and characteristic DNA fragments of Ehrlichia spp. as well as characteristic DNA fragments of Babesia spp. Amblyomma americanum, the most common tick identified, was found on a majority (56/62; 90%) of bears and accounted for 697/1,048 (66.5%) of all ticks recovered. Other ticks included Dermacentor variabilis (338/1,048; 32.3%) from 36 bears, Amblyomma maculatum (9/1,048; 0.9%) from three bears, and Ixodes scapularis (4/1,048; 0.4%) from three bears. Antibodies reactive to Ehrlichia spp. were detected in every bear tested (49/49; 100%); maximum inverse titers to Ehrlichia chaffeensis ranged from 64-4,096 (geometric mean titer 1,525). However, PCR failed to identify active infection with E. chaffeensis, Ehrlichia ewingii, or an Ehrlichia ruminantium-like agent. Infection with Babesia spp. was detected by PCR in 3/49 (6%) bears. Together these data confirm that tick infestations and infection with tick-borne disease agents are common in bears in the southern US. The significance of these infestations and infections to the health of bears, if any, and the identity of the Ehrlichia spp. responsible for the antibody reactivity seen, warrant further evaluation.

  11. Detection of human bacterial pathogens in ticks collected from Louisiana black bears (Ursus americanus luteolus).

    PubMed

    Leydet, Brian F; Liang, Fang-Ting

    2013-04-01

    There are 4 major human-biting tick species in the northeastern United States, which include: Amblyomma americanum, Amblyomma maculatum, Dermacentor variabilis, and Ixodes scapularis. The black bear is a large mammal that has been shown to be parasitized by all the aforementioned ticks. We investigated the bacterial infections in ticks collected from Louisiana black bears (Ursus americanus subspecies luteolus). Eighty-six ticks were collected from 17 black bears in Louisiana from June 2010 to March 2011. All 4 common human-biting tick species were represented. Each tick was subjected to polymerase chain reaction (PCR) targeting select bacterial pathogens and symbionts. Bacterial DNA was detected in 62% of ticks (n=53). Rickettsia parkeri, the causative agent of an emerging spotted fever group rickettsiosis, was identified in 66% of A. maculatum, 28% of D. variabilis, and 11% of I. scapularis. The Lyme disease bacterium, Borrelia burgdorferi, was detected in 2 I. scapularis, while one A. americanum was positive for Borrelia bissettii, a putative human pathogen. The rickettsial endosymbionts Candidatus Rickettsia andeanae, rickettsial endosymbiont of I. scapularis, and Rickettsia amblyommii were detected in their common tick hosts at 21%, 39%, and 60%, respectively. All ticks were PCR-negative for Anaplasma phagocytophilum, Ehrlichia spp., and Babesia microti. This is the first reported detection of R. parkeri in vector ticks in Louisiana; we also report the novel association of R. parkeri with I. scapularis. Detection of both R. parkeri and B. burgdorferi in their respective vectors in Louisiana demands further investigation to determine potential for human exposure to these pathogens. Copyright © 2013 Elsevier GmbH. All rights reserved.

  12. Modulation of gene expression in heart and liver of hibernating black bears (Ursus americanus)

    PubMed Central

    2011-01-01

    Background Hibernation is an adaptive strategy to survive in highly seasonal or unpredictable environments. The molecular and genetic basis of hibernation physiology in mammals has only recently been studied using large scale genomic approaches. We analyzed gene expression in the American black bear, Ursus americanus, using a custom 12,800 cDNA probe microarray to detect differences in expression that occur in heart and liver during winter hibernation in comparison to summer active animals. Results We identified 245 genes in heart and 319 genes in liver that were differentially expressed between winter and summer. The expression of 24 genes was significantly elevated during hibernation in both heart and liver. These genes are mostly involved in lipid catabolism and protein biosynthesis and include RNA binding protein motif 3 (Rbm3), which enhances protein synthesis at mildly hypothermic temperatures. Elevated expression of protein biosynthesis genes suggests induction of translation that may be related to adaptive mechanisms reducing cardiac and muscle atrophies over extended periods of low metabolism and immobility during hibernation in bears. Coordinated reduction of transcription of genes involved in amino acid catabolism suggests redirection of amino acids from catabolic pathways to protein biosynthesis. We identify common for black bears and small mammalian hibernators transcriptional changes in the liver that include induction of genes responsible for fatty acid β oxidation and carbohydrate synthesis and depression of genes involved in lipid biosynthesis, carbohydrate catabolism, cellular respiration and detoxification pathways. Conclusions Our findings show that modulation of gene expression during winter hibernation represents molecular mechanism of adaptation to extreme environments. PMID:21453527

  13. Behavioral Thermoregulation and Trade-Offs in Juvenile Lobster Homarus americanus.

    PubMed

    Nielsen, Travis V; McGaw, Iain J

    2016-02-01

    Water temperature influences the behavior and distribution patterns of both larval and adult American lobster Homarus americanus. However, very little is known about the responses of juvenile lobsters. The juvenile life stage is a critical period; high levels of mortality, combined with specific behavioral responses, can disconnect larval settlement from patterns of abundance of adults. We assessed behavioral thermoregulation in juvenile lobsters, and determined how thermal preferences can be altered by the presence of shelter and food. Juvenile lobsters avoided temperatures higher than 20 °C and lower than 8 °C, and had a mean temperature preference of 16.2 ± 1 °C. This preference was unaffected by prior acclimation, origin (laboratory-raised or wild), or size. When the animals were subjected to a temperature change (5-20 °C), activity rates peaked at 15 °C, and remained stable thereafter. Activity rates did not change when a shelter was added. The addition of food resulted in an increase in activity associated with food handling. When juvenile lobsters were offered a choice between temperature, shelter, and food, they always chose the environment with a shelter, even when it was in a thermally unfavorable temperature. Juveniles also spent more time in a thermally unfavorable environment when food was present; however, acquisition of a shelter was prioritized over food. Although juveniles had a similar thermal preference to adults, they are more vulnerable to predation; the innate shelter-seeking behavior of juveniles overrode their thermal preference. While temperature is an important environmental factor affecting the physiology, distribution, and growth of aquatic ectotherms, our findings suggest that trade-off behaviors occur in order to maintain optimal fitness and survival of the individual. © 2016 Marine Biological Laboratory.

  14. Soil organic matter content effects on dermal pesticide bioconcentration in American toads (Bufo americanus).

    PubMed

    Van Meter, Robin J; Glinski, Donna A; Henderson, W Matthew; Purucker, S Thomas

    2016-11-01

    Pesticides have been implicated as a major factor in global amphibian declines and may pose great risk to terrestrial phase amphibians moving to and from breeding ponds on agricultural landscapes. Dermal uptake from soil is known to occur in amphibians, but predicting pesticide availability and bioconcentration across soil types is not well understood. The present study was designed to compare uptake of 5 current-use pesticides (imidacloprid, atrazine, triadimefon, fipronil, and pendimethalin) in American toads (Bufo americanus) from exposure on soils with significant organic matter content differences (14.1% = high organic matter and 3.1% = low organic matter). We placed toads on high- or low-organic matter soil after applying individual current-use pesticides on the soil surface for an 8-h exposure duration. Whole body tissue homogenates and soils were extracted and analyzed using liquid chromatography-mass spectrometry to determine pesticide tissue and soil concentration, as well as bioconcentration factor in toads. Tissue concentrations were greater on the low-organic matter soil than the high-organic matter soil across all pesticides (average ± standard error; 1.23 ± 0.35 ppm and 0.78 ± 0.23 ppm, respectively), and bioconcentration was significantly higher for toads on the low-organic matter soil (analysis of covariance p = 0.002). Soil organic matter is known to play a significant role in the mobility of pesticides and bioavailability to living organisms. Agricultural soils typically have relatively lower organic matter content and serve as a functional habitat for amphibians. The potential for pesticide accumulation in amphibians moving throughout agricultural landscapes may be greater and should be considered in conservation and policy efforts. Environ Toxicol Chem 2016;35:2734-2741. © 2016 SETAC.

  15. Effects of simulated feeding by snow geese on Scirpus americanus rhizomes.

    PubMed

    Giroux, J -F; Bédard, J

    1987-11-01

    We simulated the feeding of Greater Snow Geese (Chen caerulescens atlantica) on the rhizomes of three-square bulrush (Scirpus americanus) in a tidal marsh along the St. Lawrence River estuary in Québec. During the spring staging period, aboveground biomass is unavailable and geese feed solely on rhizomes and overwintering buds. An experiment was designed to test the effect of three factors on subsequent growth of Scirpus: the intensity of removal (3 to 77% removal of belowground biomass), the number of "bites" (1, 2 or 3 sections removed) and the number of adventitious buds removed (1, 2 or 3). Rhizomes were dug out in May, treated and transplanted into 85-1 basins sunk in the marsh and filled with marsh soil freed of all plant material. Growth was observed weekly until the end of the growing season in August. Shoots and rhizomes were then collected, dried and weighed to obtain biomass estimates. The net above- and belowground production of Scirpus was inversely related to the initial rhizome biomass removed. At a high level of removal (>35%), the cumulative number of shoots was significantly reduced as early as two weeks after transplantation. The relative reduction in production of the treated rhizomes compared to the control plants was also related to the intensity of removal. An increased number of bites reduced production and the removal of an increased number of adventitious buds further amplified the effect of removal on rhizome production. These experimental results show that even low intensity of feeding by Snow Geese can reduce the production of Scirpus marshes.

  16. Sequential ovulation and fertility of polyoestrus in American black bears (Ursus americanus)

    PubMed Central

    Himelright, Brendan M.; Moore, Jenna M.; Gonzales, Ramona L.; Mendoza, Alejandra V.; Dye, Penny S.; Schuett, Randall J.; Durrant, Barbara S.; Read, Betsy A.; Spady, Thomas J.

    2014-01-01

    American black bears (Ursus americanus) are seasonally polyoestrous and exhibit delayed implantation, which may allow equal and independent fertility of recurrent oestruses of a mating season. We postulated that the luteal inactivity during delayed implantation allows bears to have sequential ovulation during a polyoestrous mating season such that each oestrus of a polyoestrous female will have equivalent fertility, and pregnancy would not preclude subsequent ovulation and superfetation. Controlled mating experiments were conducted on semi-free-ranging female American black bears during three mating seasons, wherein females were bred by different male cohorts in each oestrus. Behavioural observation, vulva score ranking, genetic paternity analysis, gross morphology of ovaries and microscopic morphology of diapaused embryos were used to evaluate the fertility of each subsequent oestrus in polyoestrous females. Oestrus duration, number of successful mounts and median vulva scores were similar between first and subsequent oestruses of the season. Polyoestrus occurred in 81.3% of oestrous females, with a 9.7 ± 5.5 day (mean ± SD) inter-oestrous interval. Sequential ovulation was documented in three polyoestrous females, including one that possessed both a corpus haemorrhagicum and a developed corpus luteum. Among polyoestrous dams, four of nine embryos were conceived in the first oestrus and five of nine in the second oestrus. These results indicate that each oestrus of polyoestrous females is capable of fertility, even if the female is already pregnant from a prior oestrus. Although superfetation was not directly observed in the present study, our results strongly suggest the potential of superfetation in the American black bear and provide novel insight into the complex behavioural and physiological breeding mechanisms of bears. Given that most endangered bear species share similar reproductive traits with American black bears, captive breeding programmes

  17. Plasma levels of ursodeoxycholic acid in black bears, Ursus americanus: seasonal changes.

    PubMed

    Solá, Susana; Garshelis, David L; Amaral, Joana D; Noyce, Karen V; Coy, Pam L; Steer, Clifford J; Iaizzo, Paul A; Rodrigues, Cecília M P

    2006-06-01

    To date, no other studies have examined the seasonal changes in circulating levels of various bile acids in the plasma of wild North American black bears, Ursus americanus. Using gas chromatography, bile acid concentrations were measured in plasma samples obtained during either early or late hibernation, and during summer active periods. Thus, specific compositional changes from individual animals were examined through a given year. Total bile acid concentrations in the plasma of these normal animals were found to range between 0.2 and 3.1 micromol/L (0.9 +/- 0.2 micromol/L, mean +/- SEM). Cholic, ursodeoxycholic and chenodeoxycholic acids were the major bile acid species identified. Ursodeoxycholic acid represented 28.0 +/- 2.6% of the total bile acid pool. Deoxycholic and lithocholic acids were found only in small amounts. In addition, total bile acid concentrations were lower in plasma samples obtained during hibernation compared with those obtained during summer active periods (0.6 +/- 0.1 and 1.2 +/- 0.4 micromol/L, respectively; p < 0.05). However, the relative proportion of ursodeoxycholic acid, was significantly greater in winter than in summer (31.5 +/- 3.2% and 22.2 +/- 4.5%, p < 0.05). Finally, taurine-conjugated bile acids were the predominant species in bear plasma, accounting for >67% of the total bile acids. These data demonstrate that ursodeoxycholic acid is a major bile acid in black bear plasma, mostly conjugated with taurine. Further, the finding of seasonal variation in plasma bile acid composition provides evidence to support the possible role that ursodeoxycholic acid may play in cellular protection in hibernating black bears.

  18. Blood lipid concentrations and lipoprotein patterns in captive and wild American black bears (Ursus americanus).

    PubMed

    Frank, Nicholas; Elliott, Sarah B; Allin, Shawn B; Ramsay, Edward C

    2006-02-01

    To compare blood lipid concentrations and lipoprotein patterns for captive and wild American black bears (Ursus americanus). 7 captive and 9 wild adult (> or = 4 years old) black bears. Blood was collected from 2 groups of captive black bears (groups A and B) and 1 group of wild black bears (group C). Blood triglyceride (TG) and cholesterol concentrations were compared among groups. Plasma lipoproteins were isolated by use of a self-generating gradient of iodixanol, and lipoprotein patterns were compared between groups A and B. Captive bears (mean +/- SD, 187.8 +/- 44.4 kg) weighed significantly more than wild bears (mean, 104.8 +/- 41.4 kg), but mean body weight did not differ between groups A and B. Mean blood TG concentrations for groups B (216.8 +/- 16.0 mg/dL) and C (190.7 +/- 34.0 mg/dL) were significantly higher than that of group A (103.9 +/- 25.3 mg/dL). Mean blood cholesterol concentration was also significantly higher for group B (227.8 +/- 8.2 mg/dL) than for groups A (171.7 +/- 35.5 mg/dL) or C (190.8 +/- 26.8 mg/dL). Mean very-low-density lipoprotein TG and low-density lipoprotein cholesterol concentrations were 2- and 3-fold higher, respectively, for group B, compared with concentrations for group A. Blood lipid concentrations vary significantly among populations of black bears. Plasma lipoprotein patterns of captive bears differed significantly between colonies and may have reflected differences in diet or management practices.

  19. Influence of sediment contaminated with untreated pulp and paper mill effluent on winter flounder, Pleuronectes americanus.

    PubMed

    Khan, R A

    2010-01-01

    This study was conducted to ascertain the influence of sediment contaminated with pulp and paper mill effluent in a fjord on winter flounder, Pleuronectes americanus, based on a laboratory study. Flounder, captured from a pristine site, were exposed in a flow-through system for 16 weeks to sediment collected at 2, 5, 7, and 10 km from the outfall. A group of controls was placed in uncontaminated sediment. Mortality occurred almost exclusively in fish exposed to sediment taken from 2 km than from more distant sites. Additionally, the condition factor was lower, the liver was enlarged, and toxicopathic lesions in the liver and spleen were significantly greater in fish submerged in the sediment than in fish from the more distant locations or the controls. Two ectoparasites including a ciliate, Trichodina jadranica, and a monogenean, Gyrodactylus pleuronecti, were observed only in the control group, while a digenean in the digestive tract, Steringophorus furciger, was more abundant in fish exposed to sediment from sites more distant from the outfall and the controls than at 2 km. Comparison of these results with data from a previous gradient field study on biological variables in winter flounder, captured at 2, 5, 7, and 10 km down-current from the outfall, revealed an enlarged liver that was associated with elevated levels of detoxification of hepatic enzymes and prevalence of toxicopathic lesions in both the liver and the spleen; these were significantly greater in samples taken nearest to the outfall from the mill than at more distant sites. Moreover, two metazoan parasites, S. furciger (Digenea) and Echinorhynchus gadi (Acanthocephala), in the digestive were more abundant in samples taken at farther locations and also from the reference sites. These results, based on a laboratory study, are in agreement with previous observations that winter flounder exposed to sediment at the site nearest to the outfall, where high concentrations of toxic contaminants persisted

  20. Spatio-temporal variation in serum chemistry of the lobster, Homarus americanus Milne-Edwards.

    PubMed

    Dove, A D M; Sokolowski, M S; Bartlett, S L; Bowser, P R

    2005-11-01

    Monthly variations in serum chemistry of the American lobster, Homarus americanus Milne-Edwards, were investigated at one location in Long Island Sound (LIS). Comparisons between three locations within and outside LIS were also made for a single time point. Most serum analytes displayed significant fluctuation over the study period and between locations. Temporal patterns could be classified as: low in cool months/high in warm months, i.e. Na, Cl, Na:K ratio, Ca, albumin:globulin ratio, percentage Fe saturation; high in cool months/low in warm months, i.e. pH, K, urea, total protein, albumin, globulin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), lipaemia; June spike, i.e. glucose, cholesterol, creatine kinase, iron, transferrin iron-binding capacity; other less obvious fluctuations, i.e. Mg, PO4; and no apparent fluctuation, i.e. HCO3, alkaline phosphatase. The proportion of samples correctly classified into month of collection by a subset of 13 analytes using discriminant analysis improved as the months progressed from May (0.75) to October (>0.95). Discriminant analysis also resolved 96.5% of samples by location. The significant depression of serum calcium at the eastern LIS site correlates with excretory calcinosis, a calcium storage disease described from lobsters at this site, but contrasts with a seasonal elevation in serum calcium recorded in the temporal component of the study. Serum proteins, the electrolytes Ca and K and the enzymes ALT and AST proved to have the strongest spatio-temporal patterns of variation. Serum chemistry is a useful research tool for lobster populations, but the dearth of information on the homology of analyte functions in this species with those in vertebrate species makes interpretation of the results challenging. Late summer/autumn water conditions appear to cause stress for lobsters in LIS.

  1. Vitellogenin in winter flounder (Pleuronectes americanus) from Long Island Sound and Boston Harbor

    SciTech Connect

    Pereira, J.J.; Ziskowski, J.; Mercaldo-Allen, R.; Kuropat, C.; Leudke, D.; Gould, E. )

    1992-09-01

    Vitellogenin is an egg-yolk precursor protein in teleosts which is crucial to the survival of larvae. Manufactured in the liver, where pollutants are known to accumulate, and transported to the ovary by the blood, its synthesis by the liver or uptake by the gonad can be compromised by accumulation of xenobiotics. In three studies, winter flounder (Pleuronectes americanus) blood samples were taken to determine normal levels of vitellogenin during the reproductive cycle, and to learn how its production might be affected in degraded environments. Specifically, these studies followed the seasonal cycle of vitellogenin production in winter flounder through monthly sampling at relatively clean (Shoreham, New York) and degraded areas (Black Rock and New Haven harbors, Connecticut) in Long Island Sound; examined the relationship between parental vitellogenin levels and survival of offspring by sampling fish that had been spawned at the Milford Laboratory for a reproductive success study; and determined the effect of gross liver lesions on vitellogenin production by sampling flounder from Boston Harbor, Massachusetts, which have been reported to have a high prevalence of liver tumors. Blood vitellogenin levels were determined by measuring alkali-labile phosphate (ALP). Large fish (>30 cm) from the two degraded sites had elevated serum ALP levels relative to those from the clean area. Lowered total ovarian lipid levels in large fish from Black Rock Harbor suggested impaired vitellogenin uptake. There were no significant differences in serum ALP among the small ([le]30 cm) fish from the three sampling sites. Boston Harbor flounder with gross liver lesions had lower ALP values than fish without such lesions. There were no significant differences in ALP values among the spawned fish. 43 refs., 7 figs., 6 tabs.

  2. Multidisciplinary assessment of pollution at three sites in Long Island Sound. [Pseudopleuronectes americanus

    SciTech Connect

    Gronlund, W.D.; Sinlam Chan; McCain, B.B.; Clark, R.C. Jr.; Myers, M.S.; Stein, J.E.; Brown, D.W.; Landahl, J.T.; Krahn, M.M.; Varanasi, U. )

    1991-09-01

    Winter flounder (Pseudopleuronectes americanus) were sampled from three sites located near Norwalk, New Haven, and Niantic, Connecticut, in Long Island Sound during February 1987, to evaluate the degree of chemical contamination and to determine possible effects of contaminant exposure. At each site, sediment ad infaunal invertebrates were also collected and analyzed for trace metals and organic chemicals. Specimens of liver and kidney from winter flounder were examined for histopathological conditions, including the presence of macrophage aggregates in liver tissue. Liver samples were also analyzed for DNA damage (i.e., the formation of adducts between DNA and chemical contaminants). Blood samples were collected and analyzed for erythrocyte micronuclei. The sampling site near New Haven was determined to be the most affected site, from the standpoints of greater chemical contamination and possible effects on winter flounder. Concentrations of aromatic hydrocarbons (AHs) and polychlorinated biphenyls (PCBs) were highest in sediment from this site, and the highest prevalences of the histopathological changes and DNA alterations were also found in the livers of winter flounder from this site. No differences in the concentrations of contaminants in fish or in frequencies of erythrocyte micronuclei in fish blood were found between sites. None of the sites samples had contaminant levels or prevalences of lesions as high as previously found at other East Coast locations (e.g., Boston Harbor, Massachusetts, Raritan Bay, New York). Overall, the results indicate moderate levels of pollution at two of the urban sites in Long Island Sound and provide a framework for expanded studies to better define the extent and impact of chemical pollution in Long Island Sound.

  3. Structural and Functional Characterization of a Multifunctional Alanine-Rich Peptide Analogue from Pleuronectes americanus

    PubMed Central

    Migliolo, Ludovico; Silva, Osmar N.; Silva, Paula A.; Costa, Maysa P.; Costa, Carolina R.; Nolasco, Diego O.; Barbosa, João A. R. G.; Silva, Maria R. R.; Bemquerer, Marcelo P.; Lima, Lidia M. P.; Romanos, Maria T. V.; Freitas, Sonia M.; Magalhães, Beatriz S.; Franco, Octavio L.

    2012-01-01

    Recently, defense peptides that are able to act against several targets have been characterized. The present work focuses on structural and functional evaluation of the peptide analogue Pa-MAP, previously isolated as an antifreeze peptide from Pleuronectes americanus. Pa-MAP showed activities against different targets such as tumoral cells in culture (CACO-2, MCF-7 and HCT-116), bacteria (Escherichia coli ATCC 8739 and Staphylococcus aureus ATCC 25923), viruses (HSV-1 and HSV-2) and fungi (Candida parapsilosis ATCC 22019, Trichophyton mentagrophytes (28d&E) and T. rubrum (327)). This peptide did not show toxicity against mammalian cells such as erythrocytes, Vero and RAW 264.7 cells. Molecular mechanism of action was related to hydrophobic residues, since only the terminal amino group is charged at pH 7 as confirmed by potentiometric titration. In order to shed some light on its structure-function relations, in vitro and in silico assays were carried out using circular dichroism and molecular dynamics. Furthermore, Pa-MAP showed partial unfolding of the peptide changes in a wide pH (3 to 11) and temperature (25 to 95°C) ranges, although it might not reach complete unfolding at 95°C, suggesting a high conformational stability. This peptide also showed a conformational transition with a partial α-helical fold in water and a full α-helical core in SDS and TFE environments. These results were corroborated by spectral data measured at 222 nm and by 50 ns dynamic simulation. In conclusion, data reported here show that Pa-MAP is a potential candidate for drug design against pathogenic microorganisms due to its structural stability and wide activity against a range of targets. PMID:23056574

  4. Differences in female individual reproductive potential among three stocks of winter flounder, Pseudopleuronectes americanus

    NASA Astrophysics Data System (ADS)

    McElroy, W. David; Wuenschel, Mark J.; Press, Yvonna K.; Towle, Emilee K.; McBride, Richard S.

    2013-01-01

    Potential annual fecundity (PAF) and skipped spawning of winter flounder, Pseudopleuronectes americanus, were compared among the three stocks in United States waters and between two spawning seasons. Winter flounder have group-synchronous oocyte development and determinate fecundity. These characteristics enable estimation of PAF just prior to spawning by enumeration of the late-vitellogenic cohort of oocytes, in this case employing the autodiametric method. There was a low level of down-regulation, which was limited to fish in the earlier stages of vitellogenesis. Estimates of PAF increased substantially with female size and age, ranging from < 0.5 million to > 5 million eggs per female. Fecundity at size decreased with increasing latitude. On average, fish from the Southern New England (SNE) stock had the highest individual fecundities at length and Gulf of Maine (GOM) the lowest, but differences varied among the years. Fecundity at length of fish from Georges Bank (GB) was intermediate to these two stocks and displayed less variability at size; however, GB fish grow faster so they had the highest relative fecundity at age. Skipped spawning also exhibited geographic differences; it was infrequent (< 2%) overall, but observed in the two coastal stocks (GOM more than SNE) in both years and was not observed in the GB stock. Fecundity at size between the two years was more similar for SNE fish, but all three stocks were synchronized with higher PAF in 2011 than 2010. Comparisons to previously published estimates suggest fecundity is highly variable in this species. Overall, different rates of reproductive productivity exist among individuals of the three stocks.

  5. On the Effects of Curvature on the Confinement of a Neutral Particle to a Quantum Dot Induced by Non-inertial Effects

    NASA Astrophysics Data System (ADS)

    Bakke, K.

    2012-03-01

    We discuss the influence of a linear topological defect on the bound states of a non-relativistic neutral particle with permanent magnetic dipole moment in two distinct cases: In the first case, we consider a Fermi-Walker reference frame for the observers and show how non-inertial effects yield bound states analogous to having a neutral particle subject to the Tan-Inkson model for a quantum dot (W.-C. Tan, J.C. Inkson, Semicond. Sci. Technol. 11:1635, 1996); in the second case, we consider the action of a constant force and obtain the energy levels of the bound states.

  6. Population trends and flight behavior of the American burying beetle, Nicrophorus americanus (Coleoptera: Silphidae), on Block Island, RI

    USGS Publications Warehouse

    Raithel, C.J.; Ginsberg, H.S.; Prospero, M.L.

    2006-01-01

    The endangered American burying beetle, Nicrophorus americanus, was monitored on Block Island, RI, USA, from 1991-2003 using mark-recapture population estimates of adults collected in pitfall traps. Populations increased through time, especially after 1994 when a program was initiated that provided carrion for beetle production. Beetle captures increased with increasing temperature and dew point, and decreased with increasing wind speed. Short distance movement was not related to wind direction, while longer distance flights tended to be downwind. Although many individuals flew considerable distances along transects, most recaptures were in traps near the point of release. These behaviors probably have counterbalancing effects on population estimates.

  7. Species profiles: life histories and environmental requirements of coastal fishes and invertebrates (North Atlantic). American lobster. [Homarus americanus

    SciTech Connect

    MacKenzie, C.; Moring, J.R.

    1985-04-01

    This species profile is a literature summary of the taxonomy, morphology, distribution, life history, and environmental requirements of the American lobster (Homarus americanus). These species profiles are designed to assist in the preparation of environmental impact assessments. The American lobster is a valuable commercial shellfish. After spawning, lobsters undergo a series of molts; as adults they live in coastal and offshore waters. Lobsters are captured in baited traps and incidentally in trawls. About 5 to 6 million pounds were captured commercially in 1983, a downward trend from 1971. Major environmental factors affecting reproduction, growth, and survival are water temperature, oxygen concentration, salinity, and substrate. 82 refs., 2 figs., 3 tabs.

  8. Two species of commercial flatfish, winter flounder, Pleuronectes americanus, and American plaice, Hippoglossoides platessoides, as sentinels of environmental pollution.

    PubMed

    Khan, R A

    2010-08-01

    Two species of marketable-size flatfish, winter flounder (Pleuronectes americanus) and American plaice (Hippoglossoides platessoides) from a fjord, were examined as sentinels for the effects of effluent discharged by a pulp and paper mill in Newfoundland, Canada. Reference samples for comparison were obtained from a pristine fjord. Condition factor, lymphocyte levels and the abundance of two intestinal parasites were significantly greater in flounder and plaice sampled at the reference site while macroscopic and microscopic lesions, and hepatic somatic indices were greater in flatfish taken from the contaminated site. These results suggest that both species of commercial flatfish are useful as sentinels for studying environmental pollution.

  9. Classification of Homarus americanus hemocytes and the use of differential hemocyte counts in lobsters infected with Aerococcus viridans var. homari (Gaffkemia).

    PubMed

    Battison, Andrea; Cawthorn, Richard; Horney, Barbara

    2003-11-01

    Hemocytes of the American lobster (Homarus americanus H. Milne Edwards) were classified after examination of Wright-Giemsa stained cytocentrifuge preparations by brightfield light microscopy. Eleven hemocyte types were identified using morphologic criteria. The classification system was then used to monitor changes in the differential hemocyte count (DHC) of lobsters infected with the Gram positive coccus Aerococcus viridans var. homari, etiologic agent of gaffkemia. The appearance of less mature hemocytes in the DHCs of lobsters in the late stages of infection was similar to the 'left shift' of vertebrate inflammation. Results from this study suggest that DHCs can be used to assess and characterize inflammation in H. americanus and possibly other crustaceans.

  10. Inter-animal variability in the effects of C-type allatostatin on the cardiac neuromuscular system in the lobster Homarus americanus.

    PubMed

    Wiwatpanit, Teerawat; Powers, Brian; Dickinson, Patsy S

    2012-07-01

    Although the global effects of many modulators on pattern generators are relatively consistent among preparations, modulators can induce different alterations in different preparations. We examined the mechanisms that underlie such variability in the modulatory effects of the peptide C-type allatostatin (C-AST; pQIRYHQCYFNPISCF) on the cardiac neuromuscular system of the lobster Homarus americanus. Perfusion of C-AST through the semi-intact heart consistently decreased the frequency of ongoing contractions. However, the effect of C-AST on contraction amplitude varied between preparations, decreasing in some preparations and increasing in others. To investigate this variable effect, we examined the effects of C-AST both peripherally and centrally. When contractions of the myocardium were elicited by controlled stimuli, C-AST did not alter heart contraction at the periphery (myocardium or neuromuscular junction) in any hearts. However, when applied either to the semi-intact heart or to the cardiac ganglion (CG) isolated from hearts that responded to C-AST with increased contraction force, C-AST increased both motor neuron burst duration and the number of spikes per burst by about 25%. In contrast, CG output was increased only marginally in hearts that responded to C-AST with a decrease in contraction amplitude, suggesting that the decrease in amplitude in those preparations resulted from decreased peripheral facilitation. Our data suggest that the differential effects of a single peptide on the cardiac neuromuscular system are due solely to differential effects of the peptide on the pattern generator; the extent to which the peptide induces increased burst duration is crucial in determining its overall effect on the system.

  11. Inter-animal variability in the effects of C-type allatostatin on the cardiac neuromuscular system in the lobster Homarus americanus

    PubMed Central

    Wiwatpanit, Teerawat; Powers, Brian; Dickinson, Patsy S.

    2012-01-01

    SUMMARY Although the global effects of many modulators on pattern generators are relatively consistent among preparations, modulators can induce different alterations in different preparations. We examined the mechanisms that underlie such variability in the modulatory effects of the peptide C-type allatostatin (C-AST; pQIRYHQCYFNPISCF) on the cardiac neuromuscular system of the lobster Homarus americanus. Perfusion of C-AST through the semi-intact heart consistently decreased the frequency of ongoing contractions. However, the effect of C-AST on contraction amplitude varied between preparations, decreasing in some preparations and increasing in others. To investigate this variable effect, we examined the effects of C-AST both peripherally and centrally. When contractions of the myocardium were elicited by controlled stimuli, C-AST did not alter heart contraction at the periphery (myocardium or neuromuscular junction) in any hearts. However, when applied either to the semi-intact heart or to the cardiac ganglion (CG) isolated from hearts that responded to C-AST with increased contraction force, C-AST increased both motor neuron burst duration and the number of spikes per burst by about 25%. In contrast, CG output was increased only marginally in hearts that responded to C-AST with a decrease in contraction amplitude, suggesting that the decrease in amplitude in those preparations resulted from decreased peripheral facilitation. Our data suggest that the differential effects of a single peptide on the cardiac neuromuscular system are due solely to differential effects of the peptide on the pattern generator; the extent to which the peptide induces increased burst duration is crucial in determining its overall effect on the system. PMID:22675192

  12. Ion-Ion Neutralization.

    DTIC Science & Technology

    1982-05-31

    Accession No. 3. Recipient’s Catalog Number FGL -TR-82 -0202 b- /- 4. Title (and Subtitle) 5. Type of Report & Period Covered ION-ION NEUTRALIZATION Final...few years under the terms of the grant has been the detailed study of binary ion-ion neutralization reactions involving ions of atmospheric...2TT, England. 1. INTRODUCTION Binary positive-ion negative-ion mutual neutralization viz: A+ + B->C + D (1) can be an important loss process for

  13. ALEX neutral beam probe

    SciTech Connect

    Pourrezaei, K.

    1982-01-01

    A neutral beam probe capable of measuring plasma space potential in a fully 3-dimensional magnetic field geometry has been developed. This neutral beam was successfully used to measure an arc target plasma contained within the ALEX baseball magnetic coil. A computer simulation of the experiment was performed to refine the experimental design and to develop a numerical model for scaling the ALEX neutral beam probe to other cases of fully 3-dimensional magnetic field. Based on this scaling a 30 to 50 keV neutral cesium beam probe capable of measuring space potential in the thermal barrier region of TMX Upgrade was designed.

  14. Novel hemagglutinin nanoparticle influenza vaccine with Matrix-M™ adjuvant induces hemagglutination inhibition, neutralizing, and protective responses in ferrets against homologous and drifted A(H3N2) subtypes.

    PubMed

    Smith, Gale; Liu, Ye; Flyer, David; Massare, Michael J; Zhou, Bin; Patel, Nita; Ellingsworth, Larry; Lewis, Maggie; Cummings, James F; Glenn, Greg

    2017-09-25

    Influenza viruses frequently acquire mutations undergoing antigenic drift necessitating annual evaluation of vaccine strains. Highly conserved epitopes have been identified in the hemagglutinin (HA) head and stem regions, however, current influenza vaccines induce only limited responses to these conserved sites. Here, we describe a novel seasonal recombinant HA nanoparticle influenza vaccine (NIV) formulated with a saponin-based adjuvant, Matrix-M™. NIV induced hemagglutination inhibition (HAI) and microneutralizing (MN) antibodies against a broad range of influenza A(H3N2) subtypes. In a comparison of NIV against standard-dose and high-dose inactivated influenza vaccines (IIV and IIV-HD, respectively) in ferrets NIV elicited HAI and MN responses exceeding those induced by IIV-HD against homologous A(H3N2) by 7 fold, A(H1N1) by 26 fold, and B strain viruses by 2 fold. NIV also induced MN responses against all historic A/H3N2 strains tested, spanning more than a decade of viral evolution from the 2000-2017 influenza seasons whereas IIV and IIV-HD induced HAI and MN responses were largely directed against the homologous A(H3N2), A(H1N1), and B virus strains. NIV induced superior protection compared to IIV and IIV-HD in ferrets challenged with a homologous or 10-year drifted influenza A(H3N2) strain. HAI positive and HAI negative neutralizing monoclonal antibodies derived from mice immunized with NIV were active against homologous and drifted influenza A(H3N2) strains. Taken together these observations suggest that NIV can induce responses to one or more highly conserved HA head and stem epitopes and result in highly neutralizing antibodies against both homologous and drift strains. Copyright © 2017. Published by Elsevier Ltd.

  15. The effects of ambient pH on nitrogen excretion in early life stages of the American toad (Bufo americanus).

    PubMed

    Tattersall, G J; Wright, P A

    1996-04-01

    Acidification of breeding ponds has been identified as a potential threat to the survival and health of North American amphibian populations. The effects of acid exposure on ion and acid-balance are well known, but there is little information on how environmental water pH influences nitrogen balance in amphibians. The aim of this study was to determine the effects of moderately acidic water (pH 6.0) on nitrogen excretion in early life stages of the toad, Bufo americanus. Acid exposure (pH 6.0, 54 h) resulted in a 20-80% increase in ammonia-N excretion rates in embryos and early, middle and late tadpoles stages, whereas there was no significant effect on urea-N excretion. Tissue ammonia concentrations were significantly higher (+33%) in the embryos and 35-65% lower in the three groups of tadpoles exposed to water of pH 6.0 compared to control animals (pH 8.5). In embryos, ammonia excretion accounted for greater than 90% of total nitrogen excretion (ammonia-N + urea-N), but by the late tadpole stage this value had decreased to approximately 65%. These findings indicate that exposure of embryonic and larval B. americanus to moderately acidic water disrupts nitrogen balance by increasing nitrogen loss as ammonia, with no compensatory decrease in urea excretion.

  16. Manganese concentration in lobster (Homarus americanus) gills as an index of exposure to reducing conditions in western Long Island Sound

    USGS Publications Warehouse

    Draxler, Andrew F.J.; Sherrell, Robert M.; Wieczorek, Dan; Lavigne, Michele G.; Paulson, A.J.

    2005-01-01

    We examined the accumulation of manganese (Mn) in gill tissues of chemically nai??ve lobsters held in situ at six sites in Long Island Sound (LIS) for up to six weeks to evaluate the possible contribution of eutrophication-driven habitat quality factors to the 1999 mass mortality of American lobsters (Homarus americanus). These western LIS lobster habitats experience seasonal hypoxia, which results in redox-mobilized Mn being transferred to and deposited on the tissues of the lobsters. Manganese accumulated in gill tissue of lobsters throughout the study, but rates were highest at western and southern LIS sites, ranging from 3.4-0.8 ??g/g/d (???16 ??g/g initial). The Baden-Eriksson observation that Mn accumulation in Norway lobsters (Nephrops norvegicus) is associated with ecosystem hypoxia is confirmed and extended to H. americanus. It seems likely that, after accounting for molting frequency, certain critical values may be applied to other lobster habitats of the NE US shelf. If a high proportion of lobsters in autumn have gill Mn concentrations exceeding 30 ??g/g, then the habitats are likely experiencing some reduced oxygen levels. Manganese concentrations above 100 ??g/g suggest exposure to conditions with the potential for lobster mortality should the temperatures of bottom waters become elevated, and gill concentrations above some higher level (perhaps 300 ??g/g) indicate the most severe habitat conditions with a strong potential for hypoxia stress.

  17. Molecular Detection of Ancylostoma duodenale, Ancylostoma ceylanicum, and Necator americanus in Humans in Northeastern and Southern Thailand

    PubMed Central

    Phosuk, Issarapong; Thanchomnang, Tongjit; Sanpool, Oranuch; Janwan, Penchom; Laummaunwai, Porntip; Aamnart, Witthaya; Morakote, Nimit; Maleewong, Wanchai

    2013-01-01

    The 2 principal species of hookworms infecting humans are Necator americanus and Ancylostoma duodenale. Case studies on zoonotic hookworm infections with Ancylostoma ceylanicum and/or Ancylostoma caninum are known mainly from Asian countries. Of these 2 zoonotic species, only A. ceylanicum can develop to adulthood in humans. In the present study, we report a molecular-based survey of human hookworm infections present in southern and northeastern Thailand. Thirty larval hookworm samples were obtained from fecal agar plate cultures of 10 patients in northeastren Thailand and 20 in southern Thailand. Partial ITS1, 5.8S, and ITS2 regions of the ribosomal DNA genes were amplified using PCR. The amplicons were sequenced, aligned, and compared with other hookworm sequences in GenBank database. The results showed that, in Thailand, N. americanus is more prevalent than Ancylostoma spp. and is found in both study areas. Sporadic cases of A. ceylanicum and A. duodenale infection were seen in northeastern Thailand. PMID:24516284

  18. Forces generated during stretch in the heart of the lobster Homarus americanus are anisotropic and are altered by neuromodulators

    PubMed Central

    Dickinson, E. S.; Johnson, A. S.; Ellers, O.; Dickinson, P. S.

    2016-01-01

    ABSTRACT Mechanical and neurophysiological anisotropies mediate three-dimensional responses of the heart of Homarus americanus. Although hearts in vivo are loaded multi-axially by pressure, studies of invertebrate cardiac function typically use uniaxial tests. To generate whole-heart length–tension curves, stretch pyramids at constant lengthening and shortening rates were imposed uniaxially and biaxially along longitudinal and transverse axes of the beating whole heart. To determine whether neuropeptides that are known to modulate cardiac activity in H. americanus affect the active or passive components of these length–tension curves, we also performed these tests in the presence of SGRNFLRFamide (SGRN) and GYSNRNYLRFamide (GYS). In uniaxial and biaxial tests, both passive and active forces increased with stretch along both measurement axes. The increase in passive forces was anisotropic, with greater increases along the longitudinal axis. Passive forces showed hysteresis and active forces were higher during lengthening than shortening phases of the stretch pyramid. Active forces at a given length were increased by both neuropeptides. To exert these effects, neuropeptides might have acted indirectly on the muscle via their effects on the cardiac ganglion, directly on the neuromuscular junction, or directly on the muscles. Because increases in response to stretch were also seen in stimulated motor nerve-muscle preparations, at least some of the effects of the peptides are likely peripheral. Taken together, these findings suggest that flexibility in rhythmic cardiac contractions results from the amplified effects of neuropeptides interacting with the length–tension characteristics of the heart. PMID:26896540

  19. Mass Spectral Analysis of Neuropeptide Expression and Distribution in the Nervous System of the Lobster Homarus americanus

    PubMed Central

    Chen, Ruibing; Jiang, Xiaoyue; Prieto Conaway, Maria C.; Mohtashemi, Iman; Hui, Limei; Viner, Rosa; Li, Lingjun

    2010-01-01

    The lobster Homarus americanus has long served as an important animal model for electrophysiological and behavioral studies. Using this model, we performed a comprehensive investigation of the neuropeptide expression and their localization in the nervous system, which provides useful insights for further understanding of their biological functions. Using nanoLC ESI Q-TOF MS/MS and three types of MALDI instruments, we analyzed the neuropeptide complements in a major neuroendocrine structure, pericardial organ. 57 putative neuropeptides were identified and 18 of them were de novo sequenced. Using direct tissue/extract analysis and bioinformatics software SpecPlot, we charted the global distribution of neuropeptides throughout the nervous system in H. americanus. Furthermore, we also mapped the localization of several neuropeptide families in the brain by high mass resolution and high mass accuracy mass spectrometric imaging (MSI) using a MALDI LTQ Orbitrap mass spectrometer. We have also compared the utility and instrument performance of multiple mass spectrometers for neuropeptide analysis in terms of peptidome coverage, sensitivity, mass spectral resolution and capability for de novo sequencing. PMID:20025296

  20. Dental and Temporomandibular Joint Pathology of the American Black Bear (Ursus americanus).

    PubMed

    Clark, E J; Chesnutt, S R; Winer, J N; Kass, P H; Verstraete, F J M

    Museum specimens (maxillae and/or mandibles) from 371 American black bears (Ursus americanus) acquired between 1889 and 2006 were examined macroscopically according to predefined criteria, and 348 were included in this study. Of the 348 specimens, 126 (36.2%) were from male animals, 106 (30.5%) were from female animals and 116 (33.3%) were from animals of unknown sex. Specimen ages ranged from young adult (n = 63, 18.1%) to adult (n = 285, 81.9%), with juveniles excluded from the study. The number of teeth available for examination was 12,019 (82.2%); 7.0% of teeth were absent artefactually, 0.4% were deemed absent due to acquired tooth loss and 9.7% were absent congenitally. In 43 specimens (12.3%), 82 teeth (0.68%) were small vestigial structures with crowns that were flush with the level of surrounding alveolar bone. The remaining teeth (99.3%) were of normal morphology. Only three supernumerary teeth and three instances of enamel hypoplasia were encountered. Persistent deciduous teeth or teeth with an aberrant number of roots were not encountered in any of the specimens. Approximately one-third of the teeth examined (4,543, 37.8%) displayed attrition/abrasion, affecting nearly all of the specimens (n = 338, 97.1%). Incisor and molar teeth accounted for 52.5% and 34.3% of the affected teeth, respectively, with significantly more adults affected than young adults. Dental fractures were noted in 63 bears, affecting 18.1% of specimens and 1.0% of the total number of present teeth. The canine teeth were most often fractured, with adults having significantly more complicated crown fractures of these teeth than young adults. There were 11 specimens (3.2%) that displayed periapical lesions, affecting 12 (0.1%) dental alveoli. There were 179 specimens (51.4%) displaying bony changes indicative of periodontitis, affecting 816 (6.8%) dental alveoli. The proportion of adult bears affected by periodontitis (57.9%) was significantly greater than that of young adults

  1. Polysaccharide mimicry of the epitope of the broadly neutralizing anti-HIV antibody, 2G12, induces enhanced antibody responses to self oligomannose glycans

    PubMed Central

    Dunlop, D Cameron; Bonomelli, Camille; Mansab, Fatma; Vasiljevic, Snezana; Doores, Katie J; Wormald, Mark R; Palma, Angelina S; Feizi, Ten; Harvey, David J; Dwek, Raymond A; Crispin, Max; Scanlan, Christopher N

    2010-01-01

    Immunologically, “self” carbohydrates protect the HIV-1 surface glycoprotein, gp120, from antibody recognition. However, one broadly neutralizing antibody, 2G12, neutralizes primary viral isolates by direct recognition of Manα1→2Man motifs formed by the host-derived oligomannose glycans of the viral envelope. Immunogens, capable of eliciting antibodies of similar specificity to 2G12, are therefore candidates for HIV/AIDS vaccine development. In this context, it is known that the yeast mannan polysaccharides exhibit significant antigenic mimicry with the glycans of HIV-1. Here, we report that modulation of yeast polysaccharide biosynthesis directly controls the molecular specificity of cross-reactive antibodies to self oligomannose glycans. Saccharomyces cerevisiae mannans are typically terminated by α1→3-linked mannoses that cap a Manα1→2Man motif that otherwise closely resembles the part of the oligomannose epitope recognized by 2G12. Immunization with S. cerevisiae deficient for the α1→3 mannosyltransferase gene (ΔMnn1), but not with wild-type S. cerevisiae, reproducibly elicited antibodies to the self oligomannose glycans. Carbohydrate microarray analysis of ΔMnn1 immune sera revealed fine carbohydrate specificity to Manα1→2Man units, closely matching that of 2G12. These specificities were further corroborated by enzyme-linked immunosorbent assay with chemically defined glycoforms of gp120. These antibodies exhibited remarkable similarity in the carbohydrate specificity to 2G12 and displayed statistically significant, albeit extremely weak, neutralization of HIV-1 compared to control immune sera. These data confirm the Manα1→2Man motif as the primary carbohydrate neutralization determinant of HIV-1 and show that the genetic modulation of microbial polysaccharides is a route towards immunogens capable of eliciting antibody responses to the glycans of HIV-1. PMID:20181792

  2. Immunization of mice by a multimeric L2-based linear epitope (17-36) from HPV type 16/18 induced cross reactive neutralizing antibodies.

    PubMed

    Khiavi, Farhad Motavalli; Arashkia, Arash; Nasimi, Maryam; Mahdavi, Mehdi; Golkar, Majid; Roohvand, Farzin; Azadmanesh, Kayhan

    2017-08-01

    Current licensed and commercially available prophylactic human papillomavirus (HPV) vaccines (Cervarix and quadrivalent/nine valents Gardasil) are based on major capsid protein L1 virus-like particles (VLPs) production which are expensive and type specific. Minor capsid L2-RG1 linear epitope (17-36) is a known candidate for induction of cross-neutralizing antibodies to develop low-cost pan-HPV vaccines. Herein, we report construction and expression of a three tandem repeats of L2-RG1 derived from HPV16 and 18 fused with the same head to tail pattern (HPV16:17-36×3+ HPV18:17-36×3; hereafter termed dual-type fusion L2 peptide) in E. coli and provide the results of its immunogenicity in mice. SDS-PAGE and western blot analyses indicated proper expression of the peptide that could be further purified by Ni-NTA affinity chromatography via the located C-terminal 6xHis-tag. Mice immunized by formulation of the purified peptide and Freund adjuvant raised neutralizing antibodies which showed proper cross reactivity to HPV L2 (11-200) of types: 18, 16, 31 and 45 (which totally are responsible for 90% of cervical cancers) and efficiently neutralized HPV18/16 pseudoviruses in vitro. Our results imply the possibility of development of a simple, low-cost preventive HPV vaccine based on this dual-type fusion L2 peptide in bacterial expression system with broad spectrum.

  3. Immunization of mice by a multimeric L2-based linear epitope (17-36) from HPV type 16/18 induced cross reactive neutralizing antibodies

    PubMed Central

    Khiavi, Farhad Motavalli; Arashkia, Arash; Nasimi, Maryam; Mahdavi, Mehdi; Golkar, Majid; Roohvand, Farzin; Azadmanesh, Kayhan

    2017-01-01

    Current licensed and commercially available prophylactic human papillomavirus (HPV) vaccines (Cervarix and quadrivalent/nine valents Gardasil) are based on major capsid protein L1 virus-like particles (VLPs) production which are expensive and type specific. Minor capsid L2-RG1 linear epitope (17-36) is a known candidate for induction of cross-neutralizing antibodies to develop low-cost pan-HPV vaccines. Herein, we report construction and expression of a three tandem repeats of L2-RG1 derived from HPV16 and 18 fused with the same head to tail pattern (HPV16:17-36×3+ HPV18:17-36×3; hereafter termed dual-type fusion L2 peptide) in E. coli and provide the results of its immunogenicity in mice. SDS-PAGE and western blot analyses indicated proper expression of the peptide that could be further purified by Ni-NTA affinity chromatography via the located C-terminal 6xHis-tag. Mice immunized by formulation of the purified peptide and Freund adjuvant raised neutralizing antibodies which showed proper cross reactivity to HPV L2 (11-200) of types: 18, 16, 31 and 45 (which totally are responsible for 90% of cervical cancers) and efficiently neutralized HPV18/16 pseudoviruses in vitro. Our results imply the possibility of development of a simple, low-cost preventive HPV vaccine based on this dual-type fusion L2 peptide in bacterial expression system with broad spectrum. PMID:28855937

  4. Human Papillomavirus neutralizing and cross-reactive antibodies induced in HIV-positive subjects after vaccination with quadrivalent and bivalent HPV vaccines.

    PubMed

    Faust, Helena; Toft, Lars; Sehr, Peter; Müller, Martin; Bonde, Jesper; Forslund, Ola; Østergaard, Lars; Tolstrup, Martin; Dillner, Joakim

    2016-03-18

    Ninety-one HIV-infected individuals (61 men and 30 women) were randomized to vaccination either with quadrivalent (Gardasil™) or bivalent (Cervarix™) HPV vaccine. Neutralizing and specific HPV-binding serum antibodies were measured at baseline and 12 months after the first vaccine dose. Presence of neutralizing and binding antibodies had good agreement (average Kappa for HPV types 6, 11, 16, 18, 31, 33 and 45 was 0.65). At baseline, 88% of subjects had antibodies against at least one genital HPV. Following vaccination with Cervarix™, all subjects became seropositive for HPV16 and 18. After Gardasil™ vaccination, 96% of subjects seroconverted for HPV16 and 73% for HPV18. Levels of HPV16-specific antibodies were <1 international unit (IU) in 87% of study subjects before vaccination but >10IU in 85% of study subjects after vaccination. Antibodies against non-vaccine HPV types appeared after Gardasil™ vaccination for >50% of vaccinated females for HPV 31, 35 and 73 and for >50% of Cervarix™-vaccinated females for HPV 31, 33, 35, 45, 56 and 58. Cross-reactivity with non-genital HPV types was also detected. In conclusion, HIV-infected subjects responded to HPV vaccination with induction of neutralizing antibodies against both vaccine and non-vaccine types.

  5. Evaluation of smallpox vaccines using variola neutralization.

    PubMed

    Damon, Inger K; Davidson, Whitni B; Hughes, Christine M; Olson, Victoria A; Smith, Scott K; Holman, Robert C; Frey, Sharon E; Newman, Frances; Belshe, Robert B; Yan, Lihan; Karem, Kevin

    2009-08-01

    The search for a 'third'-generation smallpox vaccine has resulted in the development and characterization of several vaccine candidates. A significant barrier to acceptance is the absence of challenge models showing induction of correlates of protective immunity against variola virus. In this light, virus neutralization provides one of few experimental methods to show specific 'in vitro' activity of vaccines against variola virus. Here, we provide characterization of the ability of a modified vaccinia virus Ankara vaccine to induce variola virus-neutralizing antibodies, and we provide comparison with the neutralization elicited by standard Dryvax vaccination.

  6. Evaluation of smallpox vaccines using variola neutralization

    PubMed Central

    Damon, Inger K.; Davidson, Whitni B.; Hughes, Christine M.; Olson, Victoria A.; Smith, Scott K.; Holman, Robert C.; Frey, Sharon E.; Newman, Frances; Belshe, Robert B.; Yan, Lihan; Karem, Kevin

    2009-01-01

    The search for a ‘third’-generation smallpox vaccine has resulted in the development and characterization of several vaccine candidates. A significant barrier to acceptance is the absence of challenge models showing induction of correlates of protective immunity against variola virus. In this light, virus neutralization provides one of few experimental methods to show specific ‘in vitro’ activity of vaccines against variola virus. Here, we provide characterization of the ability of a modified vaccinia virus Ankara vaccine to induce variola virus-neutralizing antibodies, and we provide comparison with the neutralization elicited by standard Dryvax vaccination. PMID:19339477

  7. Comparison of Two Bacterial Transport Media for Culture of Tonsilar Swabs from Bighorn Sheep ( Ovis canadensis ) and Mountain Goats ( Oreamnos americanus ).

    PubMed

    Roug, Annette; Diaz-Campos, Dubraska; Teitzel, Charlene; Besser, Thomas E

    2017-01-01

    Duplicate tonsilar swabs were collected from 77 bighorn sheep ( Ovis canadensis ) and 19 mountain goats ( Oreamnos americanus ) in Utah. Swabs were refrigerated in bacterial transport medium or frozen in cryopreservation medium prior to bacteriologic culture. The cryopreservation medium yielded comparable or superior bacterial growth while permitting more flexibility in specimen shipment to the laboratory.

  8. Use of acepromazine and medetomidine in combination for sedation and handling of Rocky Mountain elk (Cervus elaphus nelsoni) and black bears (Ursus americanus).

    PubMed

    Wolfe, Lisa L; Johnson, Heather E; Fisher, Mark C; Sirochman, Michael A; Kraft, Benjamin; Miller, Michael W

    2014-10-01

    We opportunistically evaluated a combination of acepromazine maleate and medetomidine HCl for use in sedating Rocky Mountain elk (Cervus elaphus nelsoni) and black bears (Ursus americanus) as an alternative to scheduled drug combinations. This combination was safe and effective with limitations inherent in its sedative rather than anesthetic properties.

  9. ISOLATION AND GENETIC CHARACTERIZATION OF TOXOPLASMA GONDII FROM RACCOONS (PROCYON LOTOR), CATS (FELIS DOMESTICUS), STRIPED SKUNK (MEPHITIS MEPHITIS), BLACK BEAR (URSUS AMERICANUS), AND COUGAR (PUMA CONCOLOR) FROM CANADA

    USDA-ARS?s Scientific Manuscript database

    Viable Toxoplasma gondii was isolated by bioassay in mice from tissues of 2 feral cats ( Felis domesticus), 2 raccoons (Procyon lotor), a skunk (Mephitis mephitis) trapped in remote locations in Manitoba, Canada, and a black bear (Ursus americanus ) from Kuujjuaq, northern Quebec, Canada. Geno...

  10. The introduced clam Ensis americanus in the Wadden Sea: field experiment on impact of bird predation and tidal level on survival and growth

    NASA Astrophysics Data System (ADS)

    Freudendahl, Anna Sofie L.; Nielsen, Mette M.; Jensen, Tomas; Jensen, Kurt Thomas

    2010-06-01

    In the Danish Wadden Sea the intertidal distribution of the introduced bivalve Ensis americanus (syn. E. directus) is restricted to a narrow zone around the mean low water level. To test the possible impact of birds and submersion time on dynamics and distribution of the clams, adult specimens of E. americanus collected near the low water line were transplanted to two intertidal sites and established in open and net-covered experimental plots for 9 weeks (autumn 2001). The lowest survival of clams was registered at the low-shore-site (LSS) in plots open to bird predators, suggesting that birds such as Common Eider ( Somateria mollissima) or Oystercatcher ( Haematopus ostralegus) may control the abundance of E. americanus at the lower tidal levels. For clams showing increment in shell length during the study period, the shell growth rates were highest at the LSS and lowest in the open plots at the high-shore-site (HSS). Differences in immersion time and thus food supply may explain this pattern. Body mass index (BMI) of the clams showed basically the same pattern as the survivorship: lowest BMI in open plots at the LSS and highest in the covered plots at this site. Clams from the HSS were intermediate in their BMI. Disturbance by birds in the open plots at the LSS may explain the low BMI. In conclusion birds may be an important factor controlling abundance of E. americanus in the lower intertidal zone.

  11. Measurement of nu(mu) and anti-nu(mu) induced neutral current single pi0 production cross sections on mineral oil at E(nu) ~ O(1- GeV)

    SciTech Connect

    Aguilar-Arevalo, Alexis A.; et al.

    2010-01-26

    MiniBooNE reports the first absolute cross sections for neutral current single $\\pi^0$ production on $CH_2$ induced by neutrino and antineutrino interactions measured from the largest sets of NC $\\pi^0$ events collected to date. The principal result consists of differential cross sections measured as functions of $\\pi^04$ momentum and $\\pi^0$ angle averaged over the neutrino flux at MiniBooNE. We find total cross sections of (4.76+/-0.05_{stat}+/-0.40_{sys})*$10^{-40} cm^2$/nucleon at a mean energy of <$E_\

  12. Measurement of nu(mu) and anti-nu(mu) induced neutral current single pi0 production cross sections on mineral oil at E(nu) ~ O(1- GeV)

    DOE PAGES

    Aguilar-Arevalo, Alexis A.; et al.

    2010-01-26

    MiniBooNE reports the first absolute cross sections for neutral current singlemore » $$\\pi^0$$ production on $$CH_2$$ induced by neutrino and antineutrino interactions measured from the largest sets of NC $$\\pi^0$$ events collected to date. The principal result consists of differential cross sections measured as functions of $$\\pi^04$$ momentum and $$\\pi^0$$ angle averaged over the neutrino flux at MiniBooNE. We find total cross sections of (4.76+/-0.05_{stat}+/-0.40_{sys})*$$10^{-40} cm^2$$/nucleon at a mean energy of <$$E_\

  13. Context matters: The importance of dimerization-induced conformation of the LukGH leukocidin of Staphylococcus aureus for the generation of neutralizing antibodies

    PubMed Central

    Badarau, Adriana; Rouha, Harald; Malafa, Stefan; Battles, Michael B.; Walker, Laura; Nielson, Nels; Dolezilkova, Ivana; Teubenbacher, Astrid; Banerjee, Srijib; Maierhofer, Barbara; Weber, Susanne; Stulik, Lukas; Logan, Derek T.; Welin, Martin; Mirkina, Irina; Pleban, Clara; Zauner, Gerhild; Gross, Karin; Jägerhofer, Michaela; Magyarics, Zoltán; Nagy, Eszter

    2016-01-01

    ABSTRACT LukGH (LukAB) is a potent leukocidin of Staphylococcus aureus that lyses human phagocytic cells and is thought to contribute to immune evasion. Unlike the other bi-component leukocidins of S. aureus, LukGH forms a heterodimer before binding to its receptor, CD11b expressed on professional phagocytic cells, and displays significant sequence variation. We employed a high diversity human IgG1 library presented on yeast cells to discover monoclonal antibodies (mAbs) neutralizing the cytolytic activity of LukGH. Recombinant LukG and LukH monomers or a LukGH dimer were used as capture antigens in the library selections. We found that mAbs identified with LukG or LukH as bait had no or very low toxin neutralization potency. In contrast, LukGH dimer-selected antibodies proved to be highly potent, and several mAbs were able to neutralize even the most divergent LukGH variants. Based on biolayer interferometry and mesoscale discovery, the high affinity antibody binding site on the LukGH complex was absent on the individual monomers, suggesting that it was generated upon formation of the LukG-LukH dimer. X-ray crystallography analysis of the complex between the LukGH dimer and the antigen-binding fragment of a very potent mAb (PDB code 5K59) indicated that the epitope is located in the predicted cell binding region (rim domain) of LukGH. The corresponding IgG inhibited the binding of LukGH dimer to target cells. Our data suggest that knowledge of the native conformation of target molecules is essential to generate high affinity and functional mAbs. PMID:27467113

  14. Equivalent Neutral Wind

    NASA Technical Reports Server (NTRS)

    Liu, W. Timothy; Tang, Wenqing

    1996-01-01

    The definition of equivalent neutral wind and the rationale for using it as the geophysical product of a spaceborne scatterometer are reviewed. The differences between equivalent neutral wind and actual wind, which are caused by atmospheric density stratification, are demonstrated with measurements at selected locations. A method of computing this parameter from ship and buoy measurements is described and some common fallacies in accounting for the effects of atmospheric stratification on wind shear are discussed. The computer code for the model to derive equivalent neutral wind is provided.

  15. An environmental assessment and risk map of Ascaris lumbricoides and Necator americanus distributions in Manufahi District, Timor-Leste.

    PubMed

    Wardell, Rebecca; Clements, Archie C A; Lal, Aparna; Summers, David; Llewellyn, Stacey; Campbell, Suzy J; McCarthy, James; Gray, Darren J; V Nery, Susana

    2017-05-01

    In Timor-Leste there have been intermittent and ineffective soil-transmitted helminth (STH) deworming programs since 2004. In a resource-constrained setting, having information on the geographic distribution of STH can aid in prioritising high risk communities for intervention. This study aimed to quantify the environmental risk factors for STH infection and to produce a risk map of STH in Manufahi district, Timor-Leste. Georeferenced cross-sectional data and stool samples were obtained from 2,194 participants in 606 households in 24 villages in the Manufahi District as part of cross sectional surveys done in the context of the "WASH for Worms" randomised controlled trial. Infection status was determined for Ascaris lumbricoides and Necator americanus using real-time quantitative polymerase chain reaction. Baseline infection data were linked to environmental data obtained for each household. Univariable and multivariable multilevel mixed-effects logistic regression analysis with random effects at the village and household level were conducted, with all models adjusted for age and sex. For A. lumbricoides, being a school-aged child increased the odds of infection, whilst higher temperatures in the coolest quarter of the year, alkaline soils, clay loam/loam soils and woody savannas around households were associated with decreased infection odds. For N. americanus, greater precipitation in the driest month, higher average enhanced vegetation index, age and sandy loam soils increased infection odds, whereas being female and living at higher elevations decreased the odds of infection. Predictive risk maps generated for Manufahi based upon these final models highlight the high predicted risk of N. americanus infection across the district and the more focal nature of A. lumbricoides infection. The predicted risk of any STH infection is high across the entire district. The widespread predicted risk of any STH infection in 6 to 18 year olds provides strong evidence to

  16. The influence of stray magnetic fields on ion beam neutralization

    NASA Technical Reports Server (NTRS)

    Feng, Y.-C.; Wilbur, P. J.

    1982-01-01

    An experimental investigation is described of a comparison between the ion beam neutralization characteristics of a local neutralizer (within approximately 5 cm of the beam edge) and those associated with a distant one (approximately 1 meter away from the thruster). The influence of magnetic fields in the vicinity of the neutralizer cathode orifice which are either parallel or normal to the neutralizer axis is assessed. The plasma property profiles which reflect the influence of the magnetic fields are measured. The results suggest that magnetic fields at the region of a neutralizer cathode orifice influence its ability to couple to the ion beam. They reveal that there is a potential jump from the neutralizer cathode orifice to the plasma which exists close to the orifice. This potential drop is found to increase as the axial component of magnetic flux density increases. A magnetic field perpendicular to the neutralizer axis induces a potential rise a few centimeters downstream from the neutralizer cathode.

  17. Nanoscale investigations of shift of individual interfaces in temperature induced processes of Ni-Si system by secondary neutral mass spectrometry

    NASA Astrophysics Data System (ADS)

    Lakatos, A.; Langer, G. A.; Csik, A.; Cserhati, C.; Kis-Varga, M.; Daroczi, L.; Katona, G. L.; Erdélyi, Z.; Erdelyi, G.; Vad, K.; Beke, D. L.

    2010-12-01

    We describe a method for measurement of nanoscale shift of interfaces in layered systems by a combination of secondary neutral mass spectrometry and profilometer. We demonstrate it by the example of the investigation of interface shifts during the solid state reaction in Ni/amorphous-Si system. The kinetics of the shrinkage of the initial nanocrystalline Ni film and the amorphous Si layer as well as the average growth kinetics of the product phases were determined at 503 K. The results show that nanoscale resolution can be reached and the method is promising for following solid state reactions in different thin film systems.

  18. Antitumor resistance induced by zinostatin stimalamer (ZSS), a polymer-conjugated neocarzinostatin (NCS) derivative. I. Meth A tumor eradication and tumor-neutralizing activity in mice pretreated with ZSS or NCS.

    PubMed

    Masuda, E; Maeda, H

    1995-05-01

    Zinostatin stimalamer (ZSS) is a new anticancer agent derived from neocarzinostatin (NCS), which is synthesized by conjugation of one molecule of NCS and two molecules of poly(styrene-co-maleic acid). ZSS exhibited potent in vitro and in vivo antitumor activity in preclinical experiments, and a clinical trial of the intra-arterial administration of ZSS with iodized oil on hepatocellular carcinoma showed potent antitumor activity. We investigated the effect of ZSS and NCS on antitumor resistance and found that pretreatment with either drug suppressed the growth of MethA tumors in Balb/c mice and induced tumor eradication when given separately by single administration at therapeutic doses between 1 day and 4 weeks before tumor transplantation. The findings that the cytocidal activity of these drugs was not detected in vivo at the time of tumor transplantation and that tumor regression was preceded by a period of transient growth suggested that tumor regression was due to host-mediated antitumor activity induced by these drugs. Pretreatment with ZSS or NCS also suppressed the growth of Colon 26 carcinoma and Sarcoma 180. The finding that NCS showed the same effect as ZSS suggests that poly(styrene-comaleic acid) is not essential for the induction of host-mediated antitumor activity. Furthermore, apo-ZSS, which lacks cytocidal activity, did not induce antitumor activity. From this, it is suggested that the cytocidal effect of ZSS involves the induction of host-mediated antitumor resistance. In athymic Balb/c nu/nu mice, pretreatment with ZSS or NCS did not induce tumor eradication, suggesting that mature T lymphocytes play an important role in tumor eradication. Challenging MethA was rejected without transient growth in mice that had been cured of MethA, but challenging Colon 26 was not, showing that anti-MethA resistance was augmented selectively in the MethA-eradicated mice. Splenocytes from MethA-bearing mice pretreated with the drug showed tumor-neutralizing

  19. Neutralization Assay for Chikungunya Virus Infection: Plaque Reduction Neutralization Test.

    PubMed

    Azami, Nor Azila Muhammad; Moi, Meng Ling; Takasaki, Tomohiko

    2016-01-01

    Neutralization assay is a technique that detects and quantifies neutralizing antibody in serum samples by calculating the percentage of reduction of virus activity, as the concentration of virus used is usually constant. Neutralizing antibody titer is conventionally determined by calculating the percentage reduction in total virus infectivity by counting and comparing number of plaques (localized area of infection due to cytopathic effect) with a standard amount of virus. Conventional neutralizing test uses plaque-reduction neutralization test (PRNT) to determine neutralizing antibody titers against Chikungunya virus (CHIKV). Here we describe the plaque reduction neutralization assay (PRNT) using Vero cell lines to obtain neutralizing antibody titers.

  20. Oxidant-induced formation of a neutral flavosemiquinone in the Na+-translocating NADH:Quinone oxidoreductase (Na+-NQR) from Vibrio cholerae.

    PubMed

    Tao, Minli; Casutt, Marco S; Fritz, Günter; Steuber, Julia

    2008-01-01

    The Na(+)-translocating NADH:quinone oxidoreductase (Na(+)-NQR) from the human pathogen Vibrio cholerae is a respiratory flavo-FeS complex composed of the six subunits NqrA-F. The Na(+)-NQR was produced as His(6)-tagged protein by homologous expression in V. cholerae. The isolated complex contained near-stoichiometric amounts of non-covalently bound FAD (0.78 mol/mol Na(+)-NQR) and riboflavin (0.70 mol/mol Na(+)-NQR), catalyzed NADH-driven Na(+) transport (40 nmol Na(+)min(-1) mg(-1)), and was inhibited by 2-n-heptyl-4-hydroxyquinoline-N-oxide. EPR spectroscopy showed that Na(+)-NQR as isolated contained very low amounts of a neutral flavosemiquinone (10(-3) mol/mol Na(+)-NQR). Reduction with NADH resulted in the formation of an anionic flavosemiquinone (0.10 mol/mol Na(+)-NQR). Subsequent oxidation of the Na(+)-NQR with ubiquinone-1 or O(2) led to the formation of a neutral flavosemiquinone (0.24 mol/mol Na(+)-NQR). We propose that the Na(+)-NQR is fully oxidized in its resting state, and discuss putative schemes of NADH-triggered redox transitions.

  1. Solar Neutral Particles

    NASA Image and Video Library

    This animation shows a neutral solar particle's path leaving the sun, following the magnetic field lines out to the heliosheath. The solar particle hits a hydrogen atom, stealing its electron, and ...

  2. Space station neutral external environment

    NASA Technical Reports Server (NTRS)

    Ehlers, H.; Leger, L.

    1988-01-01

    Molecular contamination levels arising from the external induced neutral environment of the Space Station (Phase 1 configuration) were calculated using the MOLFLUX model. Predicted molecular column densities and deposition rates generally meet the Space Station contamination requirements. In the doubtful cases of deposition due to materials outgassing, proper material selection, generally excluding organic products exposed to the external environment, must be considered to meet contamination requirements. It is important that the Space Station configuration, once defined, is not significantly modified to avoid introducing new unacceptable contamination sources.

  3. Ions and neutralization

    NASA Astrophysics Data System (ADS)

    Poncet, A.

    After a short presentation of intensity limitations examples due to trapped ions, the processes of ionization and neutralization build up in particle accelerators and storage rings are briefly reviewed. The tolerable limits in neutralization are then assessed at the light of current theories of incoherent and coherent effects driven by ions. Finally the usual antidotes such as clearing electrodes, missing bunch schemes and beam shaking are presented.

  4. Persistent Bovine Viral Diarrhea Virus Infection in Domestic and Wild Small Ruminants and Camelids Including the Mountain Goat (Oreamnos americanus).

    PubMed

    Nelson, Danielle D; Duprau, Jennifer L; Wolff, Peregrine L; Evermann, James F

    2015-01-01

    Bovine viral diarrhea virus (BVDV) is a pestivirus best known for causing a variety of disease syndromes in cattle, including gastrointestinal disease, reproductive insufficiency, immunosuppression, mucosal disease, and hemorrhagic syndrome. The virus can be spread by transiently infected individuals and by persistently infected animals that may be asymptomatic while shedding large amounts of virus throughout their lifetime. BVDV has been reported in over 40 domestic and free-ranging species, and persistent infection has been described in eight of those species: white-tailed deer, mule deer, eland, mousedeer, mountain goats, alpacas, sheep, and domestic swine. This paper reviews the various aspects of BVDV transmission, disease syndromes, diagnosis, control, and prevention, as well as examines BVDV infection in domestic and wild small ruminants and camelids including mountain goats (Oreamnos americanus).

  5. Persistent Bovine Viral Diarrhea Virus Infection in Domestic and Wild Small Ruminants and Camelids Including the Mountain Goat (Oreamnos americanus)

    PubMed Central

    Nelson, Danielle D.; Duprau, Jennifer L.; Wolff, Peregrine L.; Evermann, James F.

    2016-01-01

    Bovine viral diarrhea virus (BVDV) is a pestivirus best known for causing a variety of disease syndromes in cattle, including gastrointestinal disease, reproductive insufficiency, immunosuppression, mucosal disease, and hemorrhagic syndrome. The virus can be spread by transiently infected individuals and by persistently infected animals that may be asymptomatic while shedding large amounts of virus throughout their lifetime. BVDV has been reported in over 40 domestic and free-ranging species, and persistent infection has been described in eight of those species: white-tailed deer, mule deer, eland, mousedeer, mountain goats, alpacas, sheep, and domestic swine. This paper reviews the various aspects of BVDV transmission, disease syndromes, diagnosis, control, and prevention, as well as examines BVDV infection in domestic and wild small ruminants and camelids including mountain goats (Oreamnos americanus). PMID:26779126

  6. Negative density-dependent dispersal in the American black bear (Ursus americanus) revealed by noninvasive sampling and genotyping

    PubMed Central

    Roy, Justin; Yannic, Glenn; Côté, Steeve D; Bernatchez, Louis

    2012-01-01

    Although the dispersal of animals is influenced by a variety of factors, few studies have used a condition-dependent approach to assess it. The mechanisms underlying dispersal are thus poorly known in many species, especially in large mammals. We used 10 microsatellite loci to examine population density effects on sex-specific dispersal behavior in the American black bear, Ursus americanus. We tested whether dispersal increases with population density in both sexes. Fine-scale genetic structure was investigated in each of four sampling areas using Mantel tests and spatial autocorrelation analyses. Our results revealed male-biased dispersal pattern in low-density areas. As population density increased, females appeared to exhibit philopatry at smaller scales. Fine-scale genetic structure for males at higher densities may indicate reduced dispersal distances and delayed dispersal by subadults. PMID:22822432

  7. Serologic survey of Toxoplasma gondii in grizzly bears (Ursus arctos) and black bears (Ursus americanus), from Alaska, 1988 to 1991.

    PubMed

    Chomel, B B; Zarnke, R L; Kasten, R W; Kass, P H; Mendes, E

    1995-10-01

    We tested 644 serum samples from 480 grizzly bears and 40 black bears from Alaska (USA), collected between 1988 and 1991, for Toxoplasma gondii antibodies, using a commercially available latex agglutination test (LAT). A titer > or = 64 was considered positive. Serum antibody prevalence for T. gondii in grizzly bears (Ursus arctos) was 18% (87 of 480). Prevalence ranged from 9% (seven of 77) on Kodiak Island to 28% (15 of 54) in northern Alaska. Prevalence was directly correlated to age. No grizzly bears < 2-year-old had T. gondii antibody. High antibody titers were found mainly in grizzly bears captured north of the Arctic Circle. Antibody prevalence in black bears (Ursus americanus) from Interior Alaska was 15% (six of 40), similar to the prevalence in grizzly bears from the same area (13%; five of 40).

  8. Investigation of American lobster, Homarus americanus, for the presence of chlorinated dibenzo-rho-dioxins and dibenzofurans

    SciTech Connect

    Clement, R.E.; Tosine, H.M.; Taguchi, V.; Musial, C.J.; Uthe, J.F.

    1987-12-01

    The polychlorinated dibenzo-p-dioxins (PCDD) are a group of 75 compounds of current environmental concern. Most attention has been focused on the 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8,-TCDD) because of high toxicity exhibited in laboratory animal studies. Polychlorinated dibenzofurans (PCDF) comprise 135 compounds similar to the PCDD in structure and toxicity. The 2,3,7,8-tetrachlorodibenzofuran (2,3,7,8-TCDF) is the most toxic of this group. Two principal sources of PCDD and PCDF in the environment are incineration and as by-products in the manufacture and use of pentachlorophenols. American lobsters (Homarus americanus) have not been previously investigated for the presence of PCDD and PCDF. Lobsters were examined in this study because of their proximity to possible sources of PCDD and PCDF and an ability to concentrate hydrophobic organics such as polycyclic aromatic hydrocarbons (PAH) in the digestive gland.

  9. Cadmium contamination in American lobster, Homarus americanus, near a coastal lead smelter: use of multiple linear regression for management

    SciTech Connect

    Uthe, J.F.; Scott, D.P.; Chou, C.L.

    1987-04-01

    In 1980 cadmium (Cd) concentrations in digestive gland from American Lobster (Homarus americanus), captured at Belledune Harbour, New Brunswick, Canada were found to range 47.6-372 mg/kg wet weight. Since the digestive gland is commonly eaten the harbor fishery was closed. The adjacent area was decreed a controlled fishery zone. The cadmium source was a lead smelter on the harbor shore. The company installed an aqueous effluent treatment plant and improved handling procedures within the plant area. Aqueous Cd discharges were subsequently reduced by more than 95%. Cadmium levels in lobsters have been monitored annually since 1980 by sampling in early spring, immediately after the lobsters emerged from their overwintering locations. The results from the 1981-1985 monitoring program were used to model the system, analyze the data and predict the years in which certain changes to the management plan might be implemented.

  10. Serologic Survey of Snowshoe Hares (Lepus americanus) in the Greater Yellowstone Area for Brucellosis, Tularemia, and Snowshoe Hare Virus.

    PubMed

    Tyers, Dan; Zimmer, Jeremy; Lewandowski, Kristen; Hennager, Steve; Young, John; Pappert, Ryan; Panella, Amanda; Kosoy, Olga

    2015-07-01

    We examined sera from snowshoe hares (Lepus americanus) livetrapped in the northern Greater Yellowstone Area (GYA), US, for antibodies to Brucella abortus, Francisella tularensis, and snowshoe hare virus (SSHV). Zero of 90, 0 of 67, and 40 of 100 samples were antibody positive for B. abortus, F. tularensis, and SSHV, respectively. Hares were trapped from 2009 to 2012, and of the six animals that were captured twice with at least 1 yr between captures, four developed antibody to SSHV, indicating active exposure to the agent. These findings suggest snowshoe hares in the GYA do not play a significant role as a reservoir of B. abortus, but do maintain the zoonotic, encephalitic SSHV in the population.

  11. Effects of Multiple Routes of Cadmium Exposure on the Hibernation Success of the American Toad (Bufo americanus)

    USGS Publications Warehouse

    James, S.M.; Little, E.E.; Semlitsch, R.D.

    2004-01-01

    The effects of multiple routes of cadmium exposure on juvenile American toads (Bufo americanus) were evaluated using environmentally relevant concentrations. During or after exposure, toads were individually hibernated for 172 days at approximately 4??C. The following experiments were conducted: (1) dermal exposure (hibernation in soil contaminated with up to 120 ??g Cd/ g (dry weight)); (2) injection exposure (single injection with cadmium to achieve a maximum whole-body nominal concentration of 3 ??g Cd/g (wet weight) 12 days before hibernation in uncontaminated soil); and, (3) oral exposure (feeding with mealworms containing ???16 ??g Cd/g (dry weight) for 50 days before hibernation in uncontaminated soil)., We hypothesized that sublethal levels of cadmium would become lethal during hibernation because of combined chemical and cold stress. No prehibernation mortality occurred in the injection and oral exposure studies. There was a significant treatment effect on whole-body cadmium concentration in toads orally or dermally exposed and on percent of cadmium retention in toads orally exposed. There was also a trend of increased time-to-burrowing and more toads partially buried with greater cadmium concentration in the dermal study, which indicated avoidance. In all 3 experiments, no significant differences were found among cadmium treatments in hibernation survival, percent of mass loss, or locomotor performance. However, toads fed mealworms averaging 4.7 ??g Cd/g (dry weight) had only 56% survival compared with 100% survival for controls. Although our results suggest that environmentally relevant levels of cadmium do not pose a great risk to American toads, factors such as soil type or prey species may increase cadmium bioavailability, and other amphibian species may be more sensitive to cadmium than B. americanus.

  12. Phylogeographic Analyses of American Black Bears (Ursus americanus) Suggest Four Glacial Refugia and Complex Patterns of Postglacial Admixture.

    PubMed

    Puckett, Emily E; Etter, Paul D; Johnson, Eric A; Eggert, Lori S

    2015-09-01

    Studies of species with continental distributions continue to identify intraspecific lineages despite continuous habitat. Lineages may form due to isolation by distance, adaptation, divergence across barriers, or genetic drift following range expansion. We investigated lineage diversification and admixture within American black bears (Ursus americanus) across their range using 22 k single nucleotide polymorphisms and mitochondrial DNA sequences. We identified three subcontinental nuclear clusters which we further divided into nine geographic regions: Alaskan (Alaska-East), eastern (Central Interior Highlands, Great Lakes, Northeast, Southeast), and western (Alaska-West, West, Pacific Coast, Southwest). We estimated that the western cluster diverged 67 ka, before eastern and Alaskan divergence 31 ka; these divergence dates contrasted with those from the mitochondrial genome where clades A and B diverged 1.07 Ma, and clades A-east and A-west diverged 169 ka. We combined estimates of divergence timing with hindcast species distribution models to infer glacial refugia for the species in Beringia, Pacific Northwest, Southwest, and Southeast. Our results show a complex arrangement of admixture due to expansion out of multiple refugia. The delineation of the genomic population clusters was inconsistent with the ranges for 16 previously described subspecies. Ranges for U. a. pugnax and U. a. cinnamomum were concordant with admixed clusters, calling into question how to order taxa below the species level. Additionally, our finding that U. a. floridanus has not diverged from U. a. americanus also suggests that morphology and genetics should be reanalyzed to assess taxonomic designations relevant to the conservation management of the species. © The Author 2015. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution. All rights reserved.For permissions please email: journals.permissions@oup.com.

  13. Biomarker analysis of American toad (Anaxyrus americanus) and grey tree frog (Hyla versicolor) tadpoles following exposure to atrazine.

    PubMed

    Snyder, Marcía N; Henderson, W Matthew; Glinski, Donna A; Purucker, S Thomas

    2017-01-01

    The objective of the current study was to use a biomarker-based approach to investigate the influence of atrazine exposure on American toad (Anaxyrus americanus) and grey tree frog (Hyla versicolor) tadpoles. Atrazine is one of the most frequently detected herbicides in environmental matrices throughout the United States. In surface waters, it has been found at concentrations from 0.04-2859μg/L and thus presents a likely exposure scenario for non-target species such as amphibians. Studies have examined the effect of atrazine on the metamorphic parameters of amphibians, however, the data are often contradictory. Gosner stage 22-24 tadpoles were exposed to 0 (control), 10, 50, 250 or 1250μg/L of atrazine for 48h. Endogenous polar metabolites were extracted and analyzed using gas chromatography coupled with mass spectrometry. Statistical analyses of the acquired spectra with machine learning classification models demonstrated identifiable changes in the metabolomic profiles between exposed and control tadpoles. Support vector machine models with recursive feature elimination created a more efficient, non-parametric data analysis and increased interpretability of metabolomic profiles. Biochemical fluxes observed in the exposed groups of both A. americanus and H. versicolor displayed perturbations in a number of classes of biological macromolecules including fatty acids, amino acids, purine nucleosides, pyrimidines, and mono- and di-saccharides. Metabolomic pathway analyses are consistent with findings of other studies demonstrating disruption of amino acid and energy metabolism from atrazine exposure to non-target species. Copyright © 2016. Published by Elsevier B.V.

  14. Detection of serum antibodies to CagA and VacA and of serum neutralizing activity for vacuolating cytotoxin in patients with Helicobacter pylori-induced gastritis.

    PubMed Central

    Donati, M; Moreno, S; Storni, E; Tucci, A; Poli, L; Mazzoni, C; Varoli, O; Sambri, V; Farencena, A; Cevenini, R

    1997-01-01

    Thirty patients with dyspepsia, with histological diagnosis of gastritis, and with endoscopic diagnosis of peptic ulcer disease (PUD) (n = 13) or nonulcer dyspepsia (NUD) (n = 17) were admitted to the study. Helicobacter pylori vacuolating cytotoxin-producing strains (Tox+) were isolated from 14 (46.7%) patients, whereas non-cytotoxin-producing (Tox-) H. pylori strains were isolated from the remaining patients. Of 30 patients studied, 20 (66.7%) had serum cytotoxin neutralizing activity in vitro. Fourteen patients with Tox+ H. pylori strains showed serum cytotoxin neutralizing activity and serum immunoglobulin G (IgG) and IgA antibodies reactive with both 87-kDa H. pylori vacuolating cytotoxin (VacA) and 128-kDa cytotoxin-associated gene product (CagA) by immunoblotting using native enriched preparations of VacA and CagA proteins from H. pylori culture supernatants as the antigens. A 94-kDa antigen cross-reacting with the 87-kDa VacA protein could be demonstrated in culture supernatant with immune sera from humans and animals. All patients (n = 10) lacking serum neutralizing activity were also negative for IgG or IgA against VacA antigen, whereas 6 of the 10 patients showed IgG serum antibody responses against CagA antigen. The prevalence of antibodies to VacA and CagA antigens was significantly (P < 0.001) higher in patients with gastritis (20 and 26 patients for VacA and CagA, respectively, of 30 patients) than in H. pylori culture-negative controls (0 of 27 for both VacA and CagA) and in randomly selected blood donors (17 and 21 for VacA and CagA, respectively, of 120 subjects). All patients with PUD had antibodies to CagA, whereas 13 of 17 (76.5%) patients with NUD had anti-CagA antibodies. Serum IgG antibodies to VacA were present in 9 (69.2%) patients with PUD of 13 patients and in 11 (64.7%) patients with NUD of 17 patients. Anti-CagA antibodies seemed to correlate better with PUD than anti-VacA antibodies. PMID:9220168

  15. A novel hemagglutinin protein produced in bacteria protects chickens against H5N1 highly pathogenic avian influenza viruses by inducing H5 subtype-specific neutralizing antibodies

    PubMed Central

    Sączyńska, Violetta; Romanik, Agnieszka; Florys, Katarzyna; Cecuda-Adamczewska, Violetta; Kęsik-Brodacka, Małgorzata; Śmietanka, Krzysztof; Olszewska, Monika; Domańska-Blicharz, Katarzyna; Minta, Zenon; Szewczyk, Bogusław; Płucienniczak, Grażyna; Płucienniczak, Andrzej

    2017-01-01

    The highly pathogenic (HP) H5N1 avian influenza viruses (AIVs) cause a mortality rate of up to 100% in infected chickens and pose a permanent pandemic threat. Attempts to obtain effective vaccines against H5N1 HPAIVs have focused on hemagglutinin (HA), an immunodominant viral antigen capable of eliciting neutralizing antibodies. The vast majority of vaccine projects have been performed using eukaryotic expression systems. In contrast, we used a bacterial expression system to produce vaccine HA protein (bacterial HA) according to our own design. The HA protein with the sequence of the H5N1 HPAIV strain was efficiently expressed in Escherichia coli, recovered in the form of inclusion bodies and refolded by dilution between two chromatographic purification steps. Antigenicity studies showed that the resulting antigen, referred to as rH5-E. coli, preserves conformational epitopes targeted by antibodies specific for H5-subtype HAs, inhibiting hemagglutination and/or neutralizing influenza viruses in vitro. The proper conformation of this protein and its ability to form functional oligomers were confirmed by a hemagglutination test. Consistent with the biochemical characteristics, prime-boost immunizations with adjuvanted rH5-E. coli protected 100% and 70% of specific pathogen-free, layer-type chickens against challenge with homologous and heterologous H5N1 HPAIVs, respectively. The observed protection was related to the positivity in the FluAC H5 test (IDVet) but not to hemagglutination-inhibiting antibody titers. Due to full protection, the effective contact transmission of the homologous challenge virus did not occur. Survivors from both challenges did not or only transiently shed the viruses, as established by viral RNA detection in oropharyngeal and cloacal swabs. Our results demonstrate that vaccination with rH5-E. coli could confer control of H5N1 HPAIV infection and transmission rates in chicken flocks, accompanied by reduced virus shedding. Moreover, the role of

  16. Human Immunodeficiency Virus Type 1 Escape from Cyclotriazadisulfonamide-Induced CD4-Targeted Entry Inhibition Is Associated with Increased Neutralizing Antibody Susceptibility▿

    PubMed Central

    Vermeire, Kurt; Van Laethem, Kristel; Janssens, Wouter; Bell, Thomas W.; Schols, Dominique

    2009-01-01

    Continuous specific downmodulation of CD4 receptor expression in T lymphocytes by the small molecule cyclotriazadisulfonamide (CADA) selected for the CADA-resistant human immunodeficiency virus type 1 (HIV-1) NL4.3 virus containing unique mutations in the C4 and V5 regions of gp120, likely stabilizing the CD4-binding conformation. The amino acid changes in Env were associated with decreased susceptibility to anti-CD4 monoclonal antibody treatment of the cells and with higher susceptibility of the virus to soluble CD4. In addition, the acquired ability of a CADA-resistant virus to infect cells with low CD4 expression was associated with an increased susceptibility of the virus to neutralizing antibodies from sera of several HIV-1-infected patients. PMID:19570853

  17. Carboxy-terminally truncated Dengue 4 virus envelope glycoprotein expressed in Pichia pastoris induced neutralizing antibodies and resistance to Dengue 4 virus challenge in mice.

    PubMed

    Muné, M; Rodríguez, R; Ramírez, R; Soto, Y; Sierra, B; Rodríguez Roche, R; Marquez, G; Garcia, J; Guillén, G; Guzmán, M G

    2003-11-01

    We have expressed a recombinant Dengue 4 virus envelope glycoprotein (E4rec), truncated at its C-terminus by 53 amino acids, in Pichia pastoris. The presence of E4rec was confirmed by Western-blot using anti-DEN 4 hyper immune mouse ascitic fluid. E4rec migrated during SDS-PAGE as a 64 kDa protein. Treatment with endoglycosidases showed that the E protein was modified by the addition of short mannose chains and the absence of hyperglycosylation. When administered to BALB-C mice, E4rec elicited a DEN 4 neutralizing antibody response haemagglutination inhibition antibodies and specific memory T cell response. Mice immunized were also significantly protected against lethal DEN 4 virus challenge (86.6%, p < 0.001).

  18. Decreasing sensitivity to RANTES (regulated on activation, normally T cell-expressed and -secreted) neutralization of CC chemokine receptor 5-using, non-syncytium-inducing virus variants in the course of human immunodeficiency virus type 1 infection.

    PubMed

    Koning, Fransje A; Kwa, David; Boeser-Nunnink, Brigitte; Dekker, Jos; Vingerhoed, Jose; Hiemstra, Harry; Schuitemaker, Hanneke

    2003-09-15

    In approximately half of human immunodeficiency virus (HIV) type 1-infected individuals, the development of CXC chemokine receptor 4-using, syncytium-inducing (SI) virus variants precedes a rapid progression to acquired immunodeficiency syndrome (AIDS). In other individuals, only CC chemokine receptor 5-using (R5), non-SI (NSI) virus variants are present throughout infection. These individuals may be either long-term survivors (LTSs) or rapid progressors. The basis for this variable disease progression in individuals with only R5 virus variants is not yet fully understood. In this study, the beta-chemokine sensitivity of biological HIV-1 clones isolated from 13 individuals who harbored only R5, NSI virus variants (7 LTSs and 6 progressors) was investigated. We found a statistically significant decrease in sensitivity of virus variants to RANTES (regulated on activation, normally T cell-expressed and -secreted) neutralization during the course of progressive infection, but not during follow-up of LTSs. Our data suggest that a role exists for RANTES neutralization sensitivity of HIV-1 in AIDS pathogenesis.

  19. Neutral particle lithography

    NASA Astrophysics Data System (ADS)

    Craver, Barry Paul

    Neutral particle lithography (NPL) is a high resolution, proximity exposure technique where a broad beam of energetic neutral particles floods a stencil mask and transmitted beamlets transfer the mask pattern to resist on a substrate, such that each feature is printed in parallel, rather than in the serial manner of electron beam lithography. It preserves the advantages of ion beam lithography (IBL), including extremely large depth-of-field, sub-5 nm resist scattering, and the near absence of diffraction, yet is intrinsically immune to charge-related artifacts including line-edge roughness and pattern placement errors due to charge accumulation on the mask and substrate. In our experiments, a neutral particle beam is formed by passing an ion beam (e.g., 30 keV He+) through a high pressure helium gas cell (e.g., 100 mTorr) to convert the ions to energetic neutrals through charge transfer scattering. The resolution of NPL is generally superior to that of IBL for applications involving insulating substrates, large proximity gaps, and ultra-small features. High accuracy stepped exposures with energetic neutral particles, where magnetic or electrostatic deflection is impossible, have been obtained by clamping the mask to the wafer, setting the proximity gap with a suitable spacer, and mechanically inclining the mask/wafer stack relative to the beam. This approach is remarkably insensitive to vibration and thermal drift; nanometer scale image offsets have been obtained with +/-2 nm placement accuracy for experiments lasting over one hour. Using this nanostepping technique, linewidth versus dose curves were obtained, from which the NPL lithographic blur was determined as 4.4+/-1.4 nm (1sigma), which is 2-3 times smaller than the blur of electron beam lithography. Neutral particle lithography has the potential to form high density, periodic patterns with sub-10 nm resolution.

  20. Tautomerism in neutral histidine.

    PubMed

    Bermúdez, Celina; Mata, Santiago; Cabezas, Carlos; Alonso, José L

    2014-10-06

    Histidine is an important natural amino acid, involved in many relevant biological processes, which, because of its physical properties, proved difficult to characterize experimentally in its neutral form. In this work, neutral histidine has been generated in the gas phase by laser ablation of solid samples and its N(ε)H tautomeric form unraveled through its rotational spectrum. The quadrupole hyperfine structure, arising from the existing three (14)N nuclei, constituted a site-specifically probe for revealing the tautomeric form as well as the side chain configuration of this proteogenic amino acid.

  1. Lectin-like domain of thrombomodulin binds to its specific ligand Lewis Y antigen and neutralizes lipopolysaccharide-induced inflammatory response

    PubMed Central

    Shi, Chung-Sheng; Hsiao, Shi-Ming; Kao, Yuan-Chung; Kuo, Kuan-Lin; Ma, Chih-Yuan; Kuo, Cheng-Hsiang; Chang, Bi-Ing; Chang, Chuan-Fa; Lin, Chun-Hung; Wong, Chi-Huey

    2008-01-01

    Thrombomodulin (TM), a widely expressing glycoprotein originally identified in vascular endothelium, is an important cofactor in the protein C anticoagulant system. TM appears to exhibit anti-inflammatory ability through both protein C–dependent and –independent pathways. We presently have demonstrated that recombinant N-terminal lectinlike domain of TM (rTMD1) functions as a protective agent against sepsis caused by Gram-negative bacterial infections. rTMD1 caused agglutination of Escherichia coli and Klebsiella pneumoniae and enhanced the macrophage phagocytosis of these Gram-negative bacteria. Moreover, rTMD1 bound to the Klebsiella pneumoniae and lipopolysaccharide (LPS) by specifically interacting with Lewis Y antigen. rTMD1 inhibited LPS-induced inflammatory mediator production via interference with CD14 and LPS binding. Furthermore, rTMD1 modulated LPS-induced mitogen-activated protein kinase and nuclear factor-κB signaling pathway activations and inducible nitric oxide synthase expression in macrophages. Administration of rTMD1 protected the host by suppressing inflammatory responses induced by LPS and Gram-negative bacteria, and enhanced LPS and bacterial clearance in sepsis. Thus, rTMD1 can be used to defend against bacterial infection and inhibit LPS-induced inflammatory responses, suggesting that rTMD1 may be valuable in the treatment of severe inflammation in sepsis, especially in Gram-negative bacterial infections. PMID:18711002

  2. Non-neutralizing antibodies induced by seasonal influenza vaccine prevent, not exacerbate A(H1N1)pdm09 disease.

    PubMed

    Kim, Jin Hyang; Reber, Adrian J; Kumar, Amrita; Ramos, Patricia; Sica, Gabriel; Music, Nedzad; Guo, Zhu; Mishina, Margarita; Stevens, James; York, Ian A; Jacob, Joshy; Sambhara, Suryaprakash

    2016-11-16

    The association of seasonal trivalent influenza vaccine (TIV) with increased infection by 2009 pandemic H1N1 (A(H1N1)pdm09) virus, initially observed in Canada, has elicited numerous investigations on the possibility of vaccine-associated enhanced disease, but the potential mechanisms remain largely unresolved. Here, we investigated if prior immunization with TIV enhanced disease upon A(H1N1)pdm09 infection in mice. We found that A(H1N1)pdm09 infection in TIV-immunized mice did not enhance the disease, as measured by morbidity and mortality. Instead, TIV-immunized mice cleared A(H1N1)pdm09 virus and recovered at an accelerated rate compared to control mice. Prior TIV immunization was associated with potent inflammatory mediators and virus-specific CD8 T cell activation, but efficient immune regulation, partially mediated by IL-10R-signaling, prevented enhanced disease. Furthermore, in contrast to suggested pathological roles, pre-existing non-neutralizing antibodies (NNAbs) were not associated with enhanced virus replication, but rather with promoted antigen presentation through FcR-bearing cells that led to potent activation of virus-specific CD8 T cells. These findings provide new insights into interactions between pre-existing immunity and pandemic viruses.

  3. On the influence of a Rashba-type coupling induced by Lorentz-violating effects on a Landau system for a neutral particle

    SciTech Connect

    Bakke, K.; Belich, H.

    2015-03-15

    We study a possible scenario of the Lorentz symmetry violation background that allows us to build an analogue of the Landau system for a nonrelativistic Dirac neutral particle interacting with a field configuration of crossed electric and magnetic fields. We also discuss the arising of analogues of the Rashba coupling, the Zeeman term and the Darwin term from the Lorentz symmetry breaking effects, and the influence of these terms on the analogue of the Landau system confined to a two-dimensional quantum ring. Finally, we show that this analogy with the Landau system confined to a two-dimensional quantum ring allows us to establish an upper bound for the Lorentz symmetry breaking parameters. - Highlights: • Landau system from crossed electric and magnetic fields and fixed time-like vector. • Analogues of the Rashba coupling, the Zeeman term and the Darwin term. • Lorentz symmetry breaking effects on a two-dimensional quantum ring. • Upper bound for the Lorentz symmetry breaking parameters.

  4. Impact of naturally occurring variation in the human papillomavirus (HPV) 33 capsid proteins on recognition by vaccine-induced cross-neutralizing antibodies.

    PubMed

    Godi, Anna; Bissett, Sara L; Miller, Elizabeth; Beddows, Simon

    2017-07-01

    We investigated naturally occurring variation within the major (L1) and minor (L2) capsid proteins of human papillomavirus (HPV) genotype 33. Pseudoviruses (PsV) representing HPV33 lineages A1, A2, A3, B and C exhibited comparable particle-to-infectivity ratios and morphology but demonstrated a decreased sensitivity (A2, A3, B and C) to cross-neutralization by HPV vaccine antibodies compared to the A1 sublineage. Chimeric PsVs demonstrated that these differences in sensitivity were due to polymorphisms in the L1 protein, with little or no influence from variation within the L2 protein. Site-directed mutagenesis of the L1 gene identified the DE loop residue 133 and the FG residue 266 as being critical for conferring this differential sensitivity. The use of HPV33 homology models based upon the HPV16 crystal structure suggested that they are likely to act independently on more than one antibody epitope. These data improve our understanding of the potential impact of natural capsid variation on recognition by vaccine antibodies.

  5. Non-neutralizing antibodies induced by seasonal influenza vaccine prevent, not exacerbate A(H1N1)pdm09 disease

    PubMed Central

    Kim, Jin Hyang; Reber, Adrian J.; Kumar, Amrita; Ramos, Patricia; Sica, Gabriel; Music, Nedzad; Guo, Zhu; Mishina, Margarita; Stevens, James; York, Ian A.; Jacob, Joshy; Sambhara, Suryaprakash

    2016-01-01

    The association of seasonal trivalent influenza vaccine (TIV) with increased infection by 2009 pandemic H1N1 (A(H1N1)pdm09) virus, initially observed in Canada, has elicited numerous investigations on the possibility of vaccine-associated enhanced disease, but the potential mechanisms remain largely unresolved. Here, we investigated if prior immunization with TIV enhanced disease upon A(H1N1)pdm09 infection in mice. We found that A(H1N1)pdm09 infection in TIV-immunized mice did not enhance the disease, as measured by morbidity and mortality. Instead, TIV-immunized mice cleared A(H1N1)pdm09 virus and recovered at an accelerated rate compared to control mice. Prior TIV immunization was associated with potent inflammatory mediators and virus-specific CD8 T cell activation, but efficient immune regulation, partially mediated by IL-10R-signaling, prevented enhanced disease. Furthermore, in contrast to suggested pathological roles, pre-existing non-neutralizing antibodies (NNAbs) were not associated with enhanced virus replication, but rather with promoted antigen presentation through FcR-bearing cells that led to potent activation of virus-specific CD8 T cells. These findings provide new insights into interactions between pre-existing immunity and pandemic viruses. PMID:27849030

  6. Radiation-induced DNA double-strand breaks produced in histone-depleted tumor cell nuclei measured using the neutral comet assay

    SciTech Connect

    Olive, P.L.; Banath, J.P.

    1995-05-01

    Removal of histones and other nuclear proteins greatly enhances the sensitivity of mammalian cells to DNA damage by ionizing radiation. We examined the possibility that the ease of dissociation of histones, or the association of other nuclear proteins with DNA, may differ between radioresistant and sensitive human tumor cells. Cells embedded in agarose were exposed to increasing salt concentrations prior to irradiation and examination using a microscopic gel electrophoresis method, the neutral comet assay. Induction of double-strand breaks increased by a factor of about 20 when cells of four human tumor cell line HT144 melanoma, HT29 adenocarcinoma, DU145 prostate carcinoma and U87 glioma, were exposed to 2 M NaCl; however, no correlation with radiosensitivity was apparent. While a significant number of histone and non-histone proteins are present after extraction with 1.2 M NaCL, these proteins apparently have only a minor influence on radiosensitivity. However, if they are allowed to remain with DNA during electrophoresis, about 15 times more strand breaks are required to produce a similar amount of DNA migration in both DU145 and HT144 cells. These results suggest that the association between proteins and DNA within the nucleus, as probed by extraction with sodium chloride, does not help to explain differences in intrinsic radiosensitivity among cells of these diverse tumor cell lines. 33 refs., 11 figs.

  7. Application of random amplified polymorphic DNA (RAPD) analysis for identification of grouper (Epinephelus guaza), wreck fish (Polyprion americanus), and Nile perch (Lates niloticus) fillets.

    PubMed

    Asensio, Luis; González, Isabel; Fernández, Alicia; Rodríguez, Miguel A; Lobo, Esther; Hernández, Pablo E; García, Teresa; Martín, Rosario

    2002-02-01

    A random amplified polymorphic DNA (RAPD) method was developed for the specific identification of grouper (Epinephelus guaza), wreck fish (Polyprion americanus), and Nile perch (Lates niloticus) fillets. Using two different reaction primers (S1 and L1), RAPD analysis produced clear fingerprints from which the three fish species could be easily identified. This approach is rapid and reliable and offers the potential to detect fraudulent or unintentional mislabeling of these species in routine seafood authentication analysis.

  8. Cross-reactive broadly neutralizing antibodies: timing is everything.

    PubMed

    Euler, Zelda; Schuitemaker, Hanneke

    2012-01-01

    The recent surge of research into new broadly neutralizing antibodies in HIV-1 infection has recharged the field of HIV-1 vaccinology. In this review we discuss the currently known broadly neutralizing antibodies and focus on factors that may shape these antibodies in natural infection. We further discuss the role of these antibodies in the clinical course of the infection and consider immunological obstacles in inducing broadly neutralizing antibodies with a vaccine.

  9. Comparison of homologous and heterologous prime-boost vaccine approaches using Modified Vaccinia Ankara and soluble protein to induce neutralizing antibodies by the human cytomegalovirus pentamer complex in mice.

    PubMed

    Chiuppesi, Flavia; Wussow, Felix; Scharf, Louise; Contreras, Heidi; Gao, Han; Meng, Zhuo; Nguyen, Jenny; Barry, Peter A; Bjorkman, Pamela J; Diamond, Don J

    2017-01-01

    Since neutralizing antibodies (NAb) targeting the human cytomegalovirus (HCMV) pentamer complex (PC) potently block HCMV host cell entry, anti-PC NAb induction is thought to be important for a vaccine formulation to prevent HCMV infection. By developing a vaccine strategy based on soluble PC protein and using a previously generated Modified Vaccinia Ankara vector co-expressing all five PC subunits (MVA-PC), we compared HCMV NAb induction by homologous immunization using prime-boost vaccine regimen employing only PC protein or MVA-PC and heterologous immunization using prime-boost combinations of PC protein and MVA-PC. Utilizing a recently isolated anti-PC NAb, we produced highly pure soluble PC protein that displayed conformational and linear neutralizing epitopes, interfered with HCMV entry, and was recognized by antibodies induced by HCMV during natural infection. Mice vaccinated by different immunization routes with the purified PC protein in combination with a clinically approved adjuvant formulation elicited high-titer and durable HCMV NAb. While MVA-PC and soluble PC protein either alone or in combination elicited robust HCMV NAb, significantly different potencies of these vaccine approaches were observed in dependence on immunization schedule. Using only two immunizations, vaccination with MVA-PC alone or prime-boost combinations of MVA-PC and PC protein was significantly more effective in stimulating HCMV NAb than immunization with PC protein alone. In contrast, with three immunizations, NAb induced by soluble PC protein either alone or combined with two boosts of MVA-PC increased to levels that exceeded NAb titer stimulated by MVA-PC alone. These results provide insights into the potency of soluble protein and MVA to elicit NAb by the HCMV PC via homologous and heterologous prime-boost immunization, which may contribute to develop clinically deployable vaccine strategies to prevent HCMV infection.

  10. Comparison of homologous and heterologous prime-boost vaccine approaches using Modified Vaccinia Ankara and soluble protein to induce neutralizing antibodies by the human cytomegalovirus pentamer complex in mice

    PubMed Central

    Chiuppesi, Flavia; Wussow, Felix; Scharf, Louise; Contreras, Heidi; Gao, Han; Meng, Zhuo; Nguyen, Jenny; Barry, Peter A.; Bjorkman, Pamela J.

    2017-01-01

    Since neutralizing antibodies (NAb) targeting the human cytomegalovirus (HCMV) pentamer complex (PC) potently block HCMV host cell entry, anti-PC NAb induction is thought to be important for a vaccine formulation to prevent HCMV infection. By developing a vaccine strategy based on soluble PC protein and using a previously generated Modified Vaccinia Ankara vector co-expressing all five PC subunits (MVA-PC), we compared HCMV NAb induction by homologous immunization using prime-boost vaccine regimen employing only PC protein or MVA-PC and heterologous immunization using prime-boost combinations of PC protein and MVA-PC. Utilizing a recently isolated anti-PC NAb, we produced highly pure soluble PC protein that displayed conformational and linear neutralizing epitopes, interfered with HCMV entry, and was recognized by antibodies induced by HCMV during natural infection. Mice vaccinated by different immunization routes with the purified PC protein in combination with a clinically approved adjuvant formulation elicited high-titer and durable HCMV NAb. While MVA-PC and soluble PC protein either alone or in combination elicited robust HCMV NAb, significantly different potencies of these vaccine approaches were observed in dependence on immunization schedule. Using only two immunizations, vaccination with MVA-PC alone or prime-boost combinations of MVA-PC and PC protein was significantly more effective in stimulating HCMV NAb than immunization with PC protein alone. In contrast, with three immunizations, NAb induced by soluble PC protein either alone or combined with two boosts of MVA-PC increased to levels that exceeded NAb titer stimulated by MVA-PC alone. These results provide insights into the potency of soluble protein and MVA to elicit NAb by the HCMV PC via homologous and heterologous prime-boost immunization, which may contribute to develop clinically deployable vaccine strategies to prevent HCMV infection. PMID:28813507

  11. Bleach Neutralizes Mold Allergens

    ERIC Educational Resources Information Center

    Science Teacher, 2005

    2005-01-01

    Researchers at National Jewish Medical and Research Center have demonstrated that dilute bleach not only kills common household mold, but may also neutralize the mold allergens that cause most mold-related health complaints. The study, published in the Journal of Allergy and Clinical Immunology, is the first to test the effect on allergic…

  12. CO2-Neutral Fuels

    NASA Astrophysics Data System (ADS)

    Goede, Adelbert; van de Sanden, Richard

    2016-06-01

    Mimicking the biogeochemical cycle of System Earth, synthetic hydrocarbon fuels are produced from recycled CO2 and H2O powered by renewable energy. Recapturing CO2 after use closes the carbon cycle, rendering the fuel cycle CO2 neutral. Non-equilibrium molecular CO2 vibrations are key to high energy efficiency.

  13. Beyond Viral Neutralization.

    PubMed

    Lewis, George K; Pazgier, Marzena; Evans, David; Ferrari, Guido; Bournazos, Stylianos; Parsons, Matthew S; Bernard, Nicole F; Finzi, Andrés

    2017-01-13

    It has been known for more than 30 years that Human Immunodeficiency Virus 1 (HIV-1) infection drives a very potent B cell response resulting in the production of anti-HIV-1 antibodies targeting several viral proteins, particularly its envelope glycoproteins (Env). Env epitopes are exposed on the surfaces of viral particles and infected cells where they are targets of potentially protective antibodies. These antibodies can interdict infection by neutralization and there is strong evidence suggesting that Fc-mediated effector function can also contribute to protection. Current evidence suggests that Fc-mediated effector function plays a role in protection against infection by broadly neutralizing antibodies (bnAbs) and it might be important for protection by non-neutralizing antibodies. Fc-mediated effector function includes diverse mechanisms that include antibody-dependent cellular cytotoxicity (ADCC), antibody-mediated complement activation (ADC), antibody-dependent cellular phagocytosis (ADCP), antibody-dependent cell-mediated virus inhibition (ADCVI), antibody-mediated trancytosis inhibition, and antibody-mediated virus opsonization. All these functions could be beneficial in fighting viral infections including HIV-1. In this perspective, we discuss the latest developments for ADCC responses discussed at the HIVR4P satellite session on non-neutralizing antibodies, with emphasis on the mechanisms of ADCC resistance employed by HIV-1, the structural basis of epitopes recognized by antibodies that mediate ADCC, NK-cell education and ADCC, and murine models to study ADCC against HIV-1.

  14. Bleach Neutralizes Mold Allergens

    ERIC Educational Resources Information Center

    Science Teacher, 2005

    2005-01-01

    Researchers at National Jewish Medical and Research Center have demonstrated that dilute bleach not only kills common household mold, but may also neutralize the mold allergens that cause most mold-related health complaints. The study, published in the Journal of Allergy and Clinical Immunology, is the first to test the effect on allergic…

  15. Lead, chromium and manganese removal by in vitro root cultures of two aquatic macrophytes species: Typha latifolia L. and Scirpus americanus pers.

    PubMed

    Santos-Díaz, María del Socorro; Barrón-Cruz, María del Carmen

    2011-07-01

    The ability of in vitro roots cultures of Typha latifolia and Scirpus americanus to remove metals was studied. Roots were cultivated on Murashige-Skoog medium with 15 microg L(-1) Cr 11, 60 microg L(-1) Pb II or 1.8 mg L(-1) Mn II. Adsorbed metal to root surface was removed by washing with 0.042% HNO3. T. latifolia roots were able to uptake 68.8 microg Pb g(-1), 22.1 microg Cr g(-1) and 1680 microg Mn g(-1), while the S. americanus roots removed 148.3 microg Pb g(-1), 40.7 microg Cr g(-1) and 4037 microg Mn g(-1). About 80-90% of Pb and Cr were absorbed in both cultures. On the contrary, the Mn removal was due mainly to an adsorption process (82-86%). In comparison to the T. latifolia cultures, S. americanus cultures were twofold more efficient to remove Pb and Cr, and threefold more efficient to remove Mn. Both plant species capture metals in the following order: Cr >Pb >Mn. This investigation confirms that in vitro roots cultures could be an alternative as a phytoremediation approach for contaminated water with heavy metals.

  16. Can genetically modified Escherichia coli with neutral buoyancy induced by gas vesicles be used as an alternative method to clinorotation for microgravity studies?

    PubMed

    Benoit, Michael; Klaus, David

    2005-01-01

    Space flight has been shown to affect various bacterial growth parameters. It is proposed that weightlessness allows the cells to remain evenly distributed, consequently altering the chemical makeup of their surrounding fluid, and hence indirectly affecting their physiological behaviour. In support of this argument, ground-based studies using clinostats to partially simulate the quiescent environment attained in microgravity have generally been successful in producing bacterial growth characteristics that mimic responses reported under actual space conditions. A novel approach for evaluating the effects of reduced cell sedimentation is presented here through use of Escherichia coli cultures genetically modified to be neutrally buoyant. Since clinorotation would not (or would only minimally) affect cell distribution of this already near-colloidal cell system, it was hypothesized that the effects on final population density would be eliminated relative to a static control. Gas-vesicle-producing E. coli cultures were grown under clinostat and static conditions and the culture densities at 60 h were compared. As a control, E. coli that do not produce gas vesicles, but were otherwise identical to the experimental strain, were also grown under clinostat and static conditions. As hypothesized, no significant difference was observed in cell populations at 60 h between the clinorotated and static gas-vesicle-producing E. coli cultures, while the cells that did not produce gas vesicles showed a mean increase in population density of 10.5 % (P = 0.001). These results further suggest that the lack of cumulative cell sedimentation is the dominant effect of space flight on non-stirred, in vitro E. coli cultures.

  17. Modeling thermospheric neutral density

    NASA Astrophysics Data System (ADS)

    Qian, Liying

    Satellite drag prediction requires determination of thermospheric neutral density. The NCAR Thermosphere-Ionosphere-Electrodynamics General Circulation Model (TIEGCM) and the global-mean Thermosphere-Ionosphere-Mesosphere-Electrodynamics General Circulation Model (TIMEGCM) were used to quantify thermospheric neutral density and its variations, focusing on annual/semiannual variation, the effect of using measured solar irradiance on model calculations of solar-cycle variation, and global change in the thermosphere. Satellite drag data and the MSIS00 empirical model were utilized to compare to the TIEGCM simulations. The TIEGCM simulations indicated that eddy diffusion and its annual/semiannual variation is a mechanism for annual/semiannual density variation in the thermosphere. It was found that eddy diffusion near the turbopause can effectively influence thermospheric neutral density. Eddy diffusion, together with annual insolation variation and large-scale circulation, generated global annual/semiannual density variation observed by satellite drag. Using measured solar irradiance as solar input for the TIEGCM improved the solar-cycle dependency of the density calculation shown in F10.7 -based thermospheric empirical models. It has been found that the empirical models overestimate density at low solar activity. The TIEGCM simulations did not show such solar-cycle dependency. Using historic measurements of CO2 and F 10.7, simulations of the global-mean TIMEGCM showed that thermospheric neutral density at 400 km had an average long-term decrease of 1.7% per decade from 1970 to 2000. A forecast of density decrease for solar cycle 24 suggested that thermospheric density will decrease at 400 km from present to the end of solar cycle 24 at a rate of 2.7% per decade. Reduction in thermospheric density causes less atmospheric drag on earth-orbiting space objects. The implication of this long-term decrease of thermospheric neutral density is that it will increase the

  18. Seasonal patterns of winter flounder Pseudopleuronectes americanus abundance and reproductive condition on the New York Bight continental shelf.

    PubMed

    Wuenschel, M J; Able, K W; Byrne, D

    2009-05-01

    To resolve varied and sometimes conflicting accounts of spawning and habitat characteristics for winter flounder Pseudopleuronectes americanus, seasonal patterns in abundance and reproductive condition were investigated in the New York Bight, near the southern edge of their current reproductive range. Fish were collected from trawl surveys on the inner continental shelf from October 2006 to October 2007. Pseudopleuronectes americanus were most abundant during January and April surveys, were rarely collected in August, with intermediate abundances in June and October. Measurements of fish condition [hepato-somatic index (I(H)), condition factor (K) and the per cent dry mass of muscle tissue (%M(D))] and reproductive condition [gonado-somatic index (I(G))] were determined to evaluate seasonal changes in energy accumulation and depletion and reproduction. Males and females had similar patterns in body and reproductive condition, although the magnitude of change was greater for females. I(H) values were highest during spring and early summer, suggesting increased feeding following spawning. K and %M(D) increased through spring and summer then declined in the autumn and winter concurrent with gonadal development. Gonads began developing in the autumn, and in January, I(G) values approached spawning levels, with many spent individuals collected in spring. Within these general patterns, however, there was a large degree of variability among individuals, and a few mature non-reproductive ('skipped spawning') females were observed. In the period after spawning, increased energy intake, indicated by increased I(H), may influence reproductive output since this energy is gradually transferred to the muscle and used for gonadal development in the forthcoming year. The occurrence of ripening individuals on the inner continental shelf in January suggests that these fish either rapidly move into estuaries to spawn by February-March or they remain on the inner shelf to spawn, or

  19. Impact of maraviroc-resistant and low-CCR5-adapted mutations induced by in vitro passage on sensitivity to anti-envelope neutralizing antibodies.

    PubMed

    Yoshimura, Kazuhisa; Harada, Shigeyoshi; Boonchawalit, Samatchaya; Kawanami, Yoko; Matsushita, Shuzo

    2014-08-01

    The aim of this study was to generate maraviroc (MVC)-resistant viruses in vitro using a human immunodeficiency virus type 1 subtype B clinical isolate (HIV-1KP-5) to understand the mechanism(s) of resistance to MVC. To select HIV-1 variants resistant to MVC in vitro, we exposed high-chemokine (C-C motif) receptor 5 (CCR5)-expressing PM1/CCR5 cells to HIV-1KP-5 followed by serial passage in the presence of MVC. We also passaged HIV-1KP-5 in PM1 cells, which were low CCR5 expressing to determine low-CCR5-adapted substitutions and compared the Env sequences of the MVC-selected variants. Following 48 passages with MVC (10 µM), HIV-1KP-5 acquired a resistant phenotype [maximal per cent inhibition (MPI) 24%], whilst the low-CCR5-adapted variant had low sensitivity to MVC (IC50 ~200 nM), but not reduction of the MPI. The common substitutions observed in both the MVC-selected and low-CCR5-adapted variants were selected from the quasi-species, in V1, V3 and V5. After 14 passages, the MVC-selected variants harboured substitutions around the CCR5 N-terminal-binding site and V3 (V200I, T297I, K305R and M434I). The low-CCR5-adapted infectious clone became sensitive to anti-CD4bs and CD4i mAbs, but not to anti-V3 mAb and autologous plasma IgGs. Conversely, the MVC-selected clone became highly sensitive to the anti-envelope (Env) mAbs tested and the autologous plasma IgGs. These findings suggest that the four MVC-resistant mutations required for entry using MVC-bound CCR5 result in a conformational change of Env that is associated with a phenotype sensitive to anti-Env neutralizing antibodies.

  20. Short-term magnesium deficiency downregulates telomerase, upregulates neutral sphingomyelinase and induces oxidative DNA damage in cardiovascular tissues: relevance to atherogenesis, cardiovascular diseases and aging

    PubMed Central

    Shah, Nilank C; Shah, Gatha J; Li, Zhiqiang; Jiang, Xian-Cheng; Altura, Bella T; Altura, Burton M

    2014-01-01

    The present work tested the hypotheses that: 1) short-term dietary deficiency of magnesium (Mg; 21 days) in rats (MgD) would result in a downregulation of telomerase in cardiac and aortic smooth muscle cells, 2) low levels of Mg2+ added to drinking water (DW) would either prevent or greatly reduce the downregulation of telomerase in MgD, 3) MgD in rats would cause an upregulation of neutral-sphingomyelinase (N-SMAse) and p53, 4) short-term MgD would result in oxidation of DNA in diverse cardiac muscle and aortic smooth muscle cells as exemplified by measurement of 8-hydroxydeoxyguanosine (8-OH-dG), and 5) cross-talk between telomerase, N-SMase, p53, and 8-OH-dG would be evident in left ventricular (LV), right ventricular (RV), atrial and aortic smooth muscle obtained from rats subjected to short-term MgD. The data indicated that short-term MgD (10% normal dietary intake) resulted in downregulation of telomerase in LV, RV, atrial and aortic muscle cells; even very low levels of water-bourne Mg2+ (e.g., 15-40 mg/lday) either prevented or ameliorated the downregulation of telomerase. Our experiments also showed that MgD resulted in a 7-10 fold increased formation of 8-OH-dG in the cardiac and aortic muscle cells. The experiments also confirmed that short-term dietary deficiency of Mg resulted in greatly increased upregulation of N-SMAse and p53 in the cardiac and aortic muscle tissues. These new experiments point to a sizeable cross-talk among telomerase, N-SMAse, and p53 in rat cardiac and peripheral vascular muscle exposed to a short-term MgD. These studies would be compatible with the idea that even short-term MgD could cause alterations of the genome in diverse cell types leading to mutations of cardiac, vascular, and endothelial cells seen in aging and atherogenesis. Since we have shown, previously, that activation of N-SMAse in MgD leads to synthesis and release of ceramide in cardiovascular tissues and cells, we believe this pathway, most likely, helps to

  1. Pichia pastoris-expressed dengue 2 envelope forms virus-like particles without pre-membrane protein and induces high titer neutralizing antibodies.

    PubMed

    Mani, Shailendra; Tripathi, Lav; Raut, Rajendra; Tyagi, Poornima; Arora, Upasana; Barman, Tarani; Sood, Ruchi; Galav, Alka; Wahala, Wahala; de Silva, Aravinda; Swaminathan, Sathyamangalam; Khanna, Navin

    2013-01-01

    Dengue is a mosquito-borne viral disease with a global prevalence. It is caused by four closely-related dengue viruses (DENVs 1-4). A dengue vaccine that can protect against all four viruses is an unmet public health need. Live attenuated vaccine development efforts have encountered unexpected interactions between the vaccine viruses, raising safety concerns. This has emphasized the need to explore non-replicating dengue vaccine options. Virus-like particles (VLPs) which can elicit robust immunity in the absence of infection offer potential promise for the development of non-replicating dengue vaccine alternatives. We have used the methylotrophic yeast Pichia pastoris to develop DENV envelope (E) protein-based VLPs. We designed a synthetic codon-optimized gene, encoding the N-terminal 395 amino acid residues of the DENV-2 E protein. It also included 5' pre-membrane-derived signal peptide-encoding sequences to ensure proper translational processing, and 3' 6× His tag-encoding sequences to facilitate purification of the expressed protein. This gene was integrated into the genome of P. pastoris host and expressed under the alcohol oxidase 1 promoter by methanol induction. Recombinant DENV-2 protein, which was present in the insoluble membrane fraction, was extracted and purified using Ni(2+)-affinity chromatography under denaturing conditions. Amino terminal sequencing and detection of glycosylation indicated that DENV-2 E had undergone proper post-translational processing. Electron microscopy revealed the presence of discrete VLPs in the purified protein preparation after dialysis. The E protein present in these VLPs was recognized by two different conformation-sensitive monoclonal antibodies. Low doses of DENV-2 E VLPs formulated in alum were immunogenic in inbred and outbred mice eliciting virus neutralizing titers >1,1200 in flow cytometry based assays and protected AG129 mice against lethal challenge (p<0.05). The formation of immunogenic DENV-2 E VLPs in the

  2. Carbon neutral hydrocarbons.

    PubMed

    Zeman, Frank S; Keith, David W

    2008-11-13

    Reducing greenhouse gas emissions from the transportation sector may be the most difficult aspect of climate change mitigation. We suggest that carbon neutral hydrocarbons (CNHCs) offer an alternative pathway for deep emission cuts that complement the use of decarbonized energy carriers. Such fuels are synthesized from atmospheric carbon dioxide (CO2) and carbon neutral hydrogen. The result is a liquid fuel compatible with the existing transportation infrastructure and therefore capable of a gradual deployment with minimum supply disruption. Capturing the atmospheric CO2 can be accomplished using biomass or industrial methods referred to as air capture. The viability of biomass fuels is strongly dependent on the environmental impacts of biomass production. Strong constraints on land use may favour the use of air capture. We conclude that CNHCs may be a viable alternative to hydrogen or conventional biofuels and warrant a comparable level of research effort and support.

  3. Heterosis or Neutrality?

    PubMed Central

    Watterson, G. A.

    1977-01-01

    Various statistics have been proposed on an ad hoc basis to test whether alleles at a locus are selectively neutral. By considering population models in which selection operates, this paper shows that the population homozygosity is a powerful test statistic for testing departures from neutrality, in the direction of heterozygote advantage or disadvantage. The sample homozygosity plays a similar role when only sample data are available. Some numerical examples are included, showing the application of the test.—An analysis is made of the effect of heterosis on such quantities as the expected number of alleles in the population or sample, the effective number of alleles, the expected homozygosity, and on the population and sample allele frequency distributions generally. PMID:863245

  4. Antihypertensive neutral lipid

    DOEpatents

    Snyder, Fred L.; Blank, Merle L.

    1986-01-01

    The invention relates to the discovery of a class of neutral acetylated ether-linked glycerolipids having the capacity to lower blood pressure in warm-blooded animals. This physiological effect is structure sensitive requiring a long chain alkyl group at the sn-1 position and a short carbon chain acyl group (acetyl or propionyl) at the sn-2 position, and a hydroxyl group at the sn-3 position.

  5. Antihypertensive neutral lipid

    DOEpatents

    Snyder, F.L.; Blank, M.L.

    1984-10-26

    The invention relates to the discovery of a class of neutral acetylated either-linked glycerolipids having the capacity to lower blood presure in warm-blooded animals. This physiological effect is structure sensitive requiring a long chain alkyl group at the sn-1 position and a short carbon chain acyl group (acetyl or propionyl) at the sn-2 position, and a hydroxyl group at the sn-3 position.

  6. Exercise Equipment: Neutral Buoyancy

    NASA Technical Reports Server (NTRS)

    Shackelford, Linda; Valle, Paul

    2016-01-01

    Load Bearing Equipment for Neutral Buoyancy (LBE-NB) is an exercise frame that holds two exercising subjects in position as they apply counter forces to each other for lower extremity and spine loading resistance exercises. Resistance exercise prevents bone loss on ISS, but the ISS equipment is too massive for use in exploration craft. Integrating the human into the load directing, load generating, and motion control functions of the exercise equipment generates safe exercise loads with less equipment mass and volume.

  7. Neutral Buoyancy Simulator Test

    NASA Technical Reports Server (NTRS)

    1995-01-01

    A diver tests a secondary camera and maneuvering platform in Marshall's Neutral Buoyancy Simulator (NBS).The secondary camera will be beneficial for recording repairs and other extra vehicular activities (EVA) the astronuats will perform while making repairs on the Hubble Space Telescope (HST). The maneuvering platform was developed to give the astronauts something to stand on while performing maintenance tasks. These platforms were developed to be mobile so that the astronauts could move them to accommadate different sites.

  8. Neutral particle beam intensity controller

    DOEpatents

    Dagenhart, William K.

    1986-01-01

    A neutral beam intensity controller is provided for a neutral beam generator in which a neutral beam is established by accelerating ions from an ion source into a gas neutralizer. An amplitude modulated, rotating magnetic field is applied to the accelerated ion beam in the gas neutralizer to defocus the resultant neutral beam in a controlled manner to achieve intensity control of the neutral beam along the beam axis at constant beam energy. The rotating magnetic field alters the orbits of ions in the gas neutralizer before they are neutralized, thereby controlling the fraction of neutral particles transmitted out of the neutralizer along the central beam axis to a fusion device or the like. The altered path or defocused neutral particles are sprayed onto an actively cooled beam dump disposed perpendicular to the neutral beam axis and having a central open for passage of the focused beam at the central axis of the beamline. Virtually zero therough 100% intensity control is achieved by varying the magnetic field strength without altering the ion source beam intensity or its species yield.

  9. Nanogel-based pneumococcal surface protein A nasal vaccine induces microRNA-associated Th17 cell responses with neutralizing antibodies against Streptococcus pneumoniae in macaques

    PubMed