Sample records for modified live csfv

  1. Pigs immunized with a novel E2 subunit vaccine are protected from subgenotype heterologous classical swine fever virus challenge.

    PubMed

    Madera, Rachel; Gong, Wenjie; Wang, Lihua; Burakova, Yulia; Lleellish, Karen; Galliher-Beckley, Amy; Nietfeld, Jerome; Henningson, Jamie; Jia, Kaimin; Li, Ping; Bai, Jianfa; Schlup, John; McVey, Scott; Tu, Changchun; Shi, Jishu

    2016-09-09

    Classical swine fever (CSF) or hog cholera is a highly contagious swine viral disease. CSF endemic countries have to use routine vaccination with modified live virus (MLV) vaccines to prevent and control CSF. However, it is impossible to serologically differentiate MLV vaccinated pigs from those infected with CSF virus (CSFV). The aim of this study is to develop a one-dose E2-subunit vaccine that can provide protection against CSFV challenge. We hypothesize that a vaccine consisting of a suitable adjuvant and recombinant E2 with natural conformation may induce a similar level of protection as the MLV vaccine. Our experimental vaccine KNB-E2 was formulated with the recombinant E2 protein (Genotype 1.1) expressed by insect cells and an oil-in-water emulsion based adjuvant. 10 pigs (3 weeks old, 5 pigs/group) were immunized intramuscularly with one dose or two doses (3 weeks apart) KNB-E2, and 10 more control pigs were administered normal saline solution only. Two weeks after the second vaccination, all KNB-E2 vaccinated pigs and 5 control pigs were challenged with 5 × 10(5) TCID50 CSFV Honduras/1997 (Genotype 1.3, 1 ml intramuscular, 1 ml intranasal). It was found that while control pigs infected with CSFV stopped growing and developed high fever (>40 °C), high level CSFV load in blood and nasal fluid, and severe leukopenia 3-14 days post challenge, all KNB-E2 vaccinated pigs continued to grow as control pigs without CSFV exposure, did not show any fever, had low or undetectable level of CSFV in blood and nasal fluid. At the time of CSFV challenge, only pigs immunized with KNB-E2 developed high levels of E2-specific antibodies and anti-CSFV neutralizing antibodies. Our studies provide direct evidence that pigs immunized with one dose KNB-E2 can be protected clinically from CSFV challenge. This protection is likely mediated by high levels of E2-specific and anti-CSFV neutralizing antibodies.

  2. Generation and Efficacy Evaluation of a Recombinant Pseudorabies Virus Variant Expressing the E2 Protein of Classical Swine Fever Virus in Pigs.

    PubMed

    Wang, Yimin; Yuan, Jin; Cong, Xin; Qin, Hua-Yang; Wang, Chun-Hua; Li, Yongfeng; Li, Su; Luo, Yuzi; Sun, Yuan; Qiu, Hua-Ji

    2015-10-01

    Classical swine fever (CSF) is an economically important infectious disease of pigs caused by classical swine fever virus (CSFV). Pseudorabies (PR), which is caused by pseudorabies virus (PRV), is another important infectious disease of pigs and other animals. Coinfections of pigs with PRV and CSFV occur occasionally in the field. The modified live vaccine Bartha-K61 strain has played an important role in the control of PR in many countries, including China. Since late 2011, however, increasing PR outbreaks caused by an emerging PRV variant have been reported in Bartha-K61-vaccinated swine populations on many farms in China. Previously, we generated a gE/gI-deleted PRV (rPRVTJ-delgE) based on this PRV variant, which was shown to be safe and can provide rapid and complete protection against lethal challenge with the PRV variant in pigs. Here, we generated a new recombinant PRV variant expressing the E2 gene of CSFV (rPRVTJ-delgE/gI-E2) and evaluated its immunogenicity and efficacy in pigs. The results showed that rPRVTJ-delgE/gI-E2 was safe for pigs, induced detectable anti-PRV and anti-CSFV neutralizing antibodies, and provided complete protection against the lethal challenge with either the PRV TJ strain or the CSFV Shimen strain. The data indicate that rPRVTJ-delgE/gI-E2 is a promising candidate bivalent vaccine against PRV and CSFV coinfections. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  3. Efficacy of a live attenuated vaccine in classical swine fever virus postnatally persistently infected pigs.

    PubMed

    Muñoz-González, Sara; Perez-Simó, Marta; Muñoz, Marta; Bohorquez, José Alejandro; Rosell, Rosa; Summerfield, Artur; Domingo, Mariano; Ruggli, Nicolas; Ganges, Llilianne

    2015-07-09

    Classical swine fever (CSF) causes major losses in pig farming, with various degrees of disease severity. Efficient live attenuated vaccines against classical swine fever virus (CSFV) are used routinely in endemic countries. However, despite intensive vaccination programs in these areas for more than 20 years, CSF has not been eradicated. Molecular epidemiology studies in these regions suggests that the virus circulating in the field has evolved under the positive selection pressure exerted by the immune response to the vaccine, leading to new attenuated viral variants. Recent work by our group demonstrated that a high proportion of persistently infected piglets can be generated by early postnatal infection with low and moderately virulent CSFV strains. Here, we studied the immune response to a hog cholera lapinised virus vaccine (HCLV), C-strain, in six-week-old persistently infected pigs following post-natal infection. CSFV-negative pigs were vaccinated as controls. The humoral and interferon gamma responses as well as the CSFV RNA loads were monitored for 21 days post-vaccination. No vaccine viral RNA was detected in the serum samples and tonsils from CSFV postnatally persistently infected pigs for 21 days post-vaccination. Furthermore, no E2-specific antibody response or neutralising antibody titres were shown in CSFV persistently infected vaccinated animals. Likewise, no of IFN-gamma producing cell response against CSFV or PHA was observed. To our knowledge, this is the first report demonstrating the absence of a response to vaccination in CSFV persistently infected pigs.

  4. Safety of classical swine fever virus vaccine strain LOM in pregnant sows and their offspring.

    PubMed

    Lim, Seong-In; Song, Jae-Young; Kim, Jaejo; Hyun, Bang-Hun; Kim, Ha-Young; Cho, In-Soo; Kim, Byounghan; Woo, Gye-Hyeong; Lee, Jung-Bok; An, Dong-Jun

    2016-04-12

    The present study aimed to evaluate the safety of the classical swine fever virus (CSFV) vaccine strain LOM in pregnant sows. Pregnant sows with free CSFV antibody were inoculated with a commercial LOM vaccine during early pregnancy (day 38; n=3) or mid-pregnancy (days 49-59; n=11). In pregnant sows vaccinated during the early stages of gestation, abortion (day 109) was observed in one case, with two stillbirths and seven mummified fetuses. The viability of live-born piglets was 34.9% in sows vaccinated during mid-pregnancy compared with 81.8% in the control group. Post-mortem examination of the organs of the sows and piglets did not reveal any pathological lesions caused by CSFV; however, CSFV RNA was detected in the organs of several vaccinated sows and their litters. The LOM strain was transmitted from sows with free CSFV antibody to their fetus, but did not appear to induce immune tolerance in the offspring from vaccinated pregnant sows. Side effects were not observed in pregnant sows with antibody to the LOM strain: transmission from sow to their litters and stillbirth or mummified fetuses. The LOM strain may induce sterile immunity and provide rapid, long-lasting, and complete protection against CSFV; however, it should be contraindicated in pregnant sows due to potential adverse effects in pregnant sows with free CSFV antibody. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Immunogenicity in Swine of Orally Administered Recombinant Lactobacillus plantarum Expressing Classical Swine Fever Virus E2 Protein in Conjunction with Thymosin α-1 as an Adjuvant

    PubMed Central

    Xu, Yi-Gang; Guan, Xue-Ting; Liu, Zhong-Mei; Tian, Chang-Yong

    2015-01-01

    Classical swine fever, caused by classical swine fever virus (CSFV), is a highly contagious disease that results in enormous economic losses in pig industries. The E2 protein is one of the main structural proteins of CSFV and is capable of inducing CSFV-neutralizing antibodies and cytotoxic T lymphocyte (CTL) activities in vivo. Thymosin α-1 (Tα1), an immune-modifier peptide, plays a very important role in the cellular immune response. In this study, genetically engineered Lactobacillus plantarum bacteria expressing CSFV E2 protein alone (L. plantarum/pYG-E2) and in combination with Tα1 (L. plantarum/pYG-E2-Tα1) were developed, and the immunogenicity of each as an oral vaccine to induce protective immunity against CSFV in pigs was evaluated. The results showed that recombinant L. plantarum/pYG-E2 and L. plantarum/pYG-E2-Tα1 were both able to effectively induce protective immune responses in pigs against CSFV infection by eliciting immunoglobulin A (IgA)-based mucosal, immunoglobulin G (IgG)-based humoral, and CTL-based cellular immune responses via oral vaccination. Significant differences (P < 0.05) in the levels of immune responses were observed between L. plantarum/pYG-E2-Tα1 and L. plantarum/pYG-E2, suggesting a better immunogenicity of L. plantarum/pYG-E2-Tα1 as a result of the Tα1 molecular adjuvant that can enhance immune responsiveness and augment specific lymphocyte functions. Our data suggest that the recombinant Lactobacillus microecological agent expressing CSFV E2 protein combined with Tα1 as an adjuvant provides a promising strategy for vaccine development against CSFV. PMID:25819954

  6. The double-antigen ELISA concept for early detection of Erns -specific classical swine fever virus antibodies and application as an accompanying test for differentiation of infected from marker vaccinated animals.

    PubMed

    Meyer, D; Fritsche, S; Luo, Y; Engemann, C; Blome, S; Beyerbach, M; Chang, C-Y; Qiu, H-J; Becher, P; Postel, A

    2017-12-01

    Emergency vaccination with live marker vaccines represents a promising control strategy for future classical swine fever (CSF) outbreaks, and the first live marker vaccine is available in Europe. Successful implementation is dependent on a reliable accompanying diagnostic assay that allows differentiation of infected from vaccinated animals (DIVA). As induction of a protective immune response relies on virus-neutralizing antibodies against E2 protein of CSF virus (CSFV), the most promising DIVA strategy is based on detection of E rns -specific antibodies in infected swine. The aim of this study was to develop and to evaluate a novel E rns -specific prototype ELISA (pigtype CSFV E rns Ab), which may be used for CSF diagnosis including application as an accompanying discriminatory test for CSFV marker vaccines. The concept of a double-antigen ELISA was shown to be a solid strategy to detect E rns -specific antibodies against CSFV isolates of different genotypes (sensitivity: 93.5%; specificity: 99.7%). Furthermore, detection of early seroconversion is advantageous compared with a frequently used CSFV E2 antibody ELISA. Clear differences in reactivity between sera taken from infected animals and animals vaccinated with various marker vaccines were observed. In combination with the marker vaccine CP7_E2alf, the novel ELISA represents a sensitivity of 90.2% and a specificity of 93.8%. However, cross-reactivity with antibodies against ruminant pestiviruses was observed. Interestingly, the majority of samples tested false-positive in other E rns -based antibody ELISAs were identified correctly by the novel prototype E rns ELISA and vice versa. In conclusion, the pigtype CSFV E rns Ab ELISA can contribute to an improvement in routine CSFV antibody screening, particularly for analysis of sera taken at an early time point after infection and is applicable as a DIVA assay. An additional E rns antibody assay is recommended for identification of false-positive results in a pig herd immunized with the licensed CP7_E2alf marker vaccine. © 2017 Blackwell Verlag GmbH.

  7. T-cell factor-4 and MHC upregulation in pigs receiving a live attenuated classical swine fever virus (CSFV) vaccine strain with interferon-gamma adjuvant.

    PubMed

    Fan, Y-H; Lin, Y-L; Hwang, Y-C; Yang, H-C; Chiu, H-C; Chiou, S-H; Jong, M-H; Chow, K-C; Lin, C-C

    2016-10-01

    The effect of co-administration of interferon (IFN)-γ in pigs undergoing vaccination with an attenuated strain (LPC) of classical swine fever virus (CSFV) was investigated. Unvaccinated pigs demonstrated pyrexia and died 7-9 days after challenge with virulent CSFV. Pigs receiving the attenuated vaccine remained healthy after virus challenge, except for mild, transient pyrexia, whereas pigs receiving IFN-γ simultaneously with the vaccine demonstrated normal body temperatures after virus challenge. Examination by nested RT-PCR revealed greater viral load in the spleens of the pigs vaccinated with the attenuated CSFV, compared with those that had additionally received IFN-γ. Expression of major histocompatibility complex (MHC) class I and MHC class II molecules was upregulated in the spleens of the IFN-γ treated vaccinated pigs, demonstrated by immunohistochemistry. Based on Western blot analysis, anti-CSFV IgG2 antibodies were elevated in vaccinated pigs by co-administration of IFN-γ (IFN-γ(Hi): P < 0.01; IFN-γ(Lo): P <0.05). By employing the suppression subtractive hybridization technique, RT-PCR, in situ hybridization, and immunohistochemistry, T-cell factor-4 (Tcf-4) mRNA and protein expression were found to be upregulated in the spleens of vaccinated pigs that had received IFN-γ. This study suggests involvement of Tcf-4 in IFN-γ-mediated immune regulation following CSFV vaccination. Copyright © 2016 Elsevier Ltd. All rights reserved.

  8. Characterisation of vaccine-induced, broadly cross-reactive IFN-γ secreting T cell responses that correlate with rapid protection against classical swine fever virus.

    PubMed

    Graham, Simon P; Haines, Felicity J; Johns, Helen L; Sosan, Olubukola; La Rocca, S Anna; Lamp, Benjamin; Rümenapf, Till; Everett, Helen E; Crooke, Helen R

    2012-04-05

    Live attenuated C-strain classical swine fever viruses (CSFV) provide a rapid onset of protection, but the lack of a serological test that can differentiate vaccinated from infected animals limits their application in CSF outbreaks. Since immunity may precede antibody responses, we examined the kinetics and specificity of peripheral blood T cell responses from pigs vaccinated with a C-strain vaccine and challenged after five days with a genotypically divergent CSFV isolate. Vaccinated animals displayed virus-specific IFN-γ responses from day 3 post-challenge, whereas, unvaccinated challenge control animals failed to mount a detectable response. Both CD4(+) and cytotoxic CD8(+) T cells were identified as the cellular source of IFN-γ. IFN-γ responses showed extensive cross-reactivity when T cells were stimulated with CSFV isolates spanning the major genotypes. To determine the specificity of these responses, T cells were stimulated with recombinant CSFV proteins and a proteome-wide peptide library from a related virus, BVDV. Major cross-reactive peptides were mapped on the E2 and NS3 proteins. Finally, IFN-γ was shown to exert potent antiviral effects on CSFV in vitro. These data support the involvement of broadly cross-reactive T cell IFN-γ responses in the rapid protection conferred by the C-strain vaccine and this information should aid the development of the next generation of CSFV vaccines. Crown Copyright © 2012. Published by Elsevier Ltd. All rights reserved.

  9. Marker vaccine strategies and candidate CSFV marker vaccines.

    PubMed

    Dong, Xiao-Nan; Chen, Ying-Hua

    2007-01-04

    Classical swine fever (CSF) is an economically important highly contagious disease of swine worldwide. Classical swine fever virus (CSFV) is its etiological agent, and the only natural hosts are domestic pigs and wild boars. Although field CSFV strains vary in the virulence, they all result in serious losses in pig industry. Highly virulent field strains generally cause acute disease and high mortality; moderately virulent field strains raise subacute or chronic infections; postnatal infection by low virulent field strains produces subclinical infection and mortality in the new-born piglets. CSFV can cross the placental barrier, and this transplacental transmission usually results in mortality of fetuses and birth of congenitally infected pigs with a late-onset disease and death. Two main strategies to control CSF epidemic are systematic prophylactic vaccination with live attenuated vaccines (such as C-strain) and non-vaccination stamping-out policy. But neither of them is satisfying enough. Marker vaccine and companion serological diagnostic test is thought to be a promising strategy for future control and eradication of CSF. During the past 15 years, various candidate marker vaccines were constructed and evaluated in the animal experiments, including recombinant chimeric vaccines, recombinant deletion vaccines, DNA vaccines, subunit vaccines and peptide vaccines. Among them, two subunit vaccines entered the large scale marker vaccine trial of EU in 1999. Although they failed to fulfil all the demands of the Scientific Veterinary Committee, they successfully induced solid immunity against CSFV in the vaccinated pigs. It can be expected that new potent marker vaccines might be commercially available and used in systematic prophylactic vaccination campaign or emergency vaccination in the next 15 years. Here, we summarized current strategies and candidate CSFV marker vaccines. These strategies and methods are also helpful for the development of new-generation vaccines against other diseases.

  10. In Vitro Coinfection and Replication of Classical Swine Fever Virus and Porcine Circovirus Type 2 in PK15 Cells

    PubMed Central

    Zhou, Niu; Xing, Gang; Zhou, Jianwei; Jin, Yulan; Liang, Cuiqin; Gu, Jinyan; Hu, Boli; Liao, Min; Wang, Qin; Zhou, Jiyong

    2015-01-01

    Increasing clinical lines of evidence have shown the coinfection/superinfection of porcine circovirus type 2 (PCV2) and classical swine fever virus (CSFV). Here, we investigated whether PCV2 and CSFV could infect the same cell productively by constructing an in vitro coinfection model. Our results indicated that PCV2-free PK15 cells but not ST cells were more sensitive to PCV2, and the PK15 cell line could stably harbor replicating CSFV (PK15-CSFV cells) with a high infection rate. Confocal and super-resolution microscopic analysis showed that PCV2 and CSFV colocalized in the same PK15-CSFV cell, and the CSFV E2 protein translocated from the cytoplasm to the nucleus in PK15-CSFV cells infected with PCV2. Moreover, PCV2-CSFV dual-positive cells increased gradually in PK15-CSFV cells in a PCV2 dose-dependent manner. In PK15-CSFV cells, PCV2 replicated well, and the production of PCV2 progeny was not influenced by CSFV infection. However, CSFV reproduction decreased in a PCV2 dose-dependent manner. In addition, cellular apoptosis was not strengthened in PK15-CSFV cells infected with PCV2 in comparison with PCV2-infected PK15 cells. Moreover, using this coinfection model we further demonstrated PCV2-induced apoptosis might contribute to the impairment of CSFV HCLV strain replication in coinfected cells. Taken together, our results demonstrate for the first time the coinfection/superinfection of PCV2 and CSFV within the same cell, providing an in vitro model to facilitate further investigation of the underlying mechanism of CSFV and PCV2 coinfection. PMID:26431319

  11. A novel alphavirus replicon-vectored vaccine delivered by adenovirus induces sterile immunity against classical swine fever.

    PubMed

    Sun, Yuan; Li, Hong-Yu; Tian, Da-Yong; Han, Qiu-Ying; Zhang, Xin; Li, Na; Qiu, Hua-Ji

    2011-10-26

    Low efficacy of gene-based vaccines due to inefficient gene delivery and expression has been major bottleneck of their applications. Efforts have been made to improve the efficacy, such as gene gun and electroporation, but the strategies are difficult to put into practical use. In this study, we developed and evaluated an adenovirus-delivered, alphavirus replicon-vectored vaccine (chimeric vector-based vaccine) expressing the E2 gene of classical swine fever virus (CSFV) (rAdV-SFV-E2). Rabbits immunized with rAdV-SFV-E2 developed CSFV-specific antibodies as early as 9 days and as long as 189 days and completely protected from challenge with C-strain. Pigs immunized with rAdV-SFV-E2 (n=5) developed robust humoral and cell-mediated responses to CSFV and were completely protected from subsequent lethal CSFV infection clinically and virologically. The level of immunity and protection induced by rAdV-SFV-E2 was comparable to that provided by the currently used live attenuated vaccine, C-strain. In contrast, both the conventional alphavirus replicon-vectored vaccine pSFV1CS-E2 and conventional adenovirus-vectored vaccine rAdV-E2 provided incomplete protection. The chimeric vector-based vaccine represents the first gene-based vaccine that is able to confer sterile immunity and complete protection against CSFV. The new-concept vaccination strategy may also be valuable in vaccine development against other pathogens. Copyright © 2011 Elsevier Ltd. All rights reserved.

  12. CSFV induced mitochondrial fission and mitophagy to inhibit apoptosis

    PubMed Central

    Xu, Hailuan; Yuan, Jin; He, Wencheng; Zhu, Mengjiao; Ding, Hongxing; Yi, Lin; Chen, Jinding

    2017-01-01

    Classical swine fever virus (CSFV), which causes typical clinical characteristics in piglets, including hemorrhagic syndrome and immunosuppression, is linked to hepatitis C and dengue virus. Oxidative stress and a reduced mitochondrial transmembrane potential are disturbed in CSFV-infected cells. The balance of mitochondrial dynamics is essential for cellular homeostasis. In this study, we offer the first evidence that CSFV induces mitochondrial fission and mitophagy to inhibit host cell apoptosis for persistent infection. The formation of mitophagosomes and decline in mitochondrial mass relevant to mitophagy were detected in CSFV-infected cells. CSFV infection increased the expression and mitochondrial translocation of Pink and Parkin. Upon activation of the PINK1 and Parkin pathways, Mitofusin 2 (MFN2), a mitochondrial fusion mediator, was ubiquitinated and degraded in CSFV-infected cells. Mitophagosomes and mitophagolysosomes induced by CSFV were, respectively, observed by the colocalization of LC3-associated mitochondria with Parkin or lysosomes. In addition, a sensitive dual fluorescence reporter (mito-mRFP-EGFP) was utilized to analyze the delivery of mitophagosomes to lysosomes. Mitochondrial fission caused by CSFV infection was further determined by mitochondrial fragmentation and Drp1 translocation into mitochondria using a confocal microscope. The preservation of mitochondrial proteins, upregulated apoptotic signals and decline of viral replication resulting from the silencing of Drp1 and Parkin in CSFV-infected cells suggested that CSFV induced mitochondrial fission and mitophagy to enhance cell survival and viral persistence. Our data for mitochondrial fission and selective mitophagy in CSFV-infected cells reveal a unique view of the pathogenesis of CSFV infection and provide new avenues for the development of antiviral strategies. PMID:28455958

  13. The challenges of classical swine fever control: modified live and E2 subunit vaccines.

    PubMed

    Huang, Yu-Liang; Deng, Ming-Chung; Wang, Fun-In; Huang, Chin-Cheng; Chang, Chia-Yi

    2014-01-22

    Classical swine fever (CSF) is an economically important, highly contagious disease of swine worldwide. CSF is caused by classical swine fever virus (CSFV), and domestic pigs and wild boars are its only natural hosts. The two main strategies used to control CSF epidemic are systematic prophylactic vaccination and a non-vaccination stamping-out policy. This review compares the protective efficacy of the routinely used modified live vaccine (MLV) and E2 subunit vaccines and summarizes the factors that influence the efficacy of the vaccines and the challenges that both vaccines face to CSF control. Although MLV provide earlier and more complete protection than E2 subunit vaccines, it has the drawback of not allowing differentiation between infected and vaccinated animals (DIVA). The marker vaccine of E2 protein with companion discriminatory test to detect antibodies against E(rns) allows DIVA and is a promising strategy for future control and eradication of CSF. Maternal derived antibody (MDA) is the critical factor in impairing the efficacy of both MLV and E2 subunit vaccines, so the well-designed vaccination programs of sows and piglets should be considered together. Because of the antigen variation among various genotypes of CSFV, antibodies raised by either MLV or subunit vaccine neutralize genotypically homologous strains better than heterologous ones. However, although this is not a major concern for MLV as the induced immune responses can protect pigs against the challenge of various genotypes of CSFVs, it is critical for E2 subunit vaccines. It is thus necessary to evaluate whether the E2 subunit vaccine can completely protect against the current prevalent strains in the field. An ideal new generation of vaccine should be able to maintain the high protective efficiency of MLV and overcome the problem of antigenic variations while allowing for DIVA. Copyright © 2013 Elsevier B.V. All rights reserved.

  14. A Crystal Structure of Classical Swine Fever Virus NS5B Reveals a Novel N-terminal Domain.

    PubMed

    Li, Weiwei; Wu, Baixing; Soca, Wibowo Adian; An, Lei

    2018-05-02

    Classical swine fever virus (CSFV) is the ringleader of Classical swine fever (CSF). The non-structural protein 5B (NS5B) encodes an RNA-dependent RNA polymerase (RdRp) that is a key enzyme initiating viral RNA replication by a de novo mechanism. It is also an attractive target for the development of anti-CSFV drugs. To gain a better understanding on the mechanism of CSFV RNA synthesis, here we solved the first crystal structure of CSFV-NS5B. Our studies show that the CSFV-NS5B RdRp contains characteristic fingers, palm domain and thumb domain as well as a unique N-terminal domain (NTD) that had never been observed. Mutagenesis studies on NS5B validated the importance of NTD in the catalytic activity of this novel RNA-dependent RNA polymerase. Moreover, our results shed light on the understanding of CSFV infection. IMPORTANCE Pigs are important domestic animal. However, a highly contagious viral disease named Classical swine fever (CSF) causes devastating economic losses. Classical swine fever virus (CSFV) is the primary culprit of CSF, which is a positive-sense single-stranded RNA virus belonging to the Pestivirus genus, Flaviviridae family. Genome replication of CSFV depends on RNA-dependent RNA polymerase known as NS5B. However, the structure of CSFV-NS5B has never been reported, and the mechanism of CSFV replication is poorly understood. Here, we solved the first crystal structure of CSFV-NS5B, analyzed the function of characteristic fingers, palm, and thumb domains. Additionally, our structure also revealed the presence of a novel N-terminal domain (NTD). Biochemical studies demonstrated that the NTD of CSFV-NS5B is very important for RNA-dependent RNA polymerase (RdRp) activity. Collectively, our studies provide a structural basis for future rational design of anti-CSFV drugs which is critically important as no effective anti-CSFV drugs have been developed. Copyright © 2018 American Society for Microbiology.

  15. Chimeric classical swine fever (CSF)-Japanese encephalitis (JE) viral replicon as a non-transmissible vaccine candidate against CSF and JE infections.

    PubMed

    Yang, Zhenhua; Wu, Rui; Li, Robert W; Li, Ling; Xiong, Zhongliang; Zhao, Haizhong; Guo, Deyin; Pan, Zishu

    2012-04-01

    A trans-complemented chimeric CSF-JE virus replicon was constructed using an infectious cDNA clone of the CSF virus (CSFV) Alfort/187 strain. The CSFV E2 gene was deleted, and a fragment containing the region encoding a truncated envelope protein (tE, amino acid 292-402, domain III) of JE virus (JEV) was inserted into the resultant plasmid, pA187delE2, to generate the recombinant cDNA clone pA187delE2/JEV-tE. Porcine kidney 15 (PK15) cells that constitutively express the CSFV E2p7 proteins were then transfected with in vitro-transcribed RNA from pA187delE2/JEV-tE. As a result, the chimeric CSF-JE virus replicon particle (VRP), rv187delE2/JEV-tE, was rescued. In a mouse model, immunization with the chimeric CSF-JE VRP induced strong production of JEV-specific antibody and conferred protection against a lethal JEV challenge. Pigs immunized with CSF-JE VRP displayed strong anti-CSFV and anti-JEV antibody responses and protection against CSFV and JEV challenge infections. Our evidence suggests that E2-complemented CSF-JE VRP not only has potential as a live-attenuated non-transmissible vaccine candidate against CSF and JE but also serves as a potential DIVA (Differentiating Infected from Vaccinated Animals) vaccine for CSF in pigs. Together, our data suggest that the non-transmissible chimeric VRP expressing foreign antigenic proteins may represent a promising strategy for bivalent DIVA vaccine design. Copyright © 2012 Elsevier B.V. All rights reserved.

  16. Tumor Necrosis Factor Receptor-Associated Factor 5 Interacts with the NS3 Protein and Promotes Classical Swine Fever Virus Replication.

    PubMed

    Lv, Huifang; Dong, Wang; Guo, Kangkang; Jin, Mingxing; Li, Xiaomeng; Li, Cunfa; Zhang, Yanming

    2018-06-05

    Classical swine fever, caused by classical swine fever virus (CSFV), is a highly contagious and high-mortality viral disease, causing huge economic losses in the swine industry worldwide. CSFV non-structural protein 3 (NS3), a multifunctional protein, plays crucial roles in viral replication. However, how NS3 exactly exerts these functions is currently unknown. Here, we identified tumor necrosis factor receptor-associated factor 5 (TRAF5) as a novel binding partner of the NS3 protein via yeast two-hybrid, co-immunoprecipitation and glutathione S -transferase pull-down assays. Furthermore, we observed that TRAF5 promoted CSFV replication in porcine alveolar macrophages (PAMs). Additionally, CSFV infection or NS3 expression upregulated TRAF5 expression, implying that CSFV may exploit TRAF5 via NS3 for better growth. Moreover, CSFV infection and TRAF5 expression activated p38 mitogen activated protein kinase (MAPK) activity, and inhibition of p38 MAPK activation by the SB203580 inhibitor suppressed CSFV replication. Notably, TRAF5 overexpression did not promote CSFV replication following inhibition of p38 MAPK activation. Our findings reveal that TRAF5 promotes CSFV replication via p38 MAPK activation. This work provides a novel insight into the role of TRAF5 in CSFV replication capacity.

  17. Interferon-Inducible Oligoadenylate Synthetase-Like Protein Acts as an Antiviral Effector against Classical Swine Fever Virus via the MDA5-Mediated Type I Interferon-Signaling Pathway.

    PubMed

    Li, Lian-Feng; Yu, Jiahui; Zhang, Yuexiu; Yang, Qian; Li, Yongfeng; Zhang, Lingkai; Wang, Jinghan; Li, Su; Luo, Yuzi; Sun, Yuan; Qiu, Hua-Ji

    2017-06-01

    Classical swine fever virus (CSFV) is the causative agent of classical swine fever (CSF), which poses a serious threat to the global pig industry. Interferons (IFNs) and IFN-stimulated genes (ISGs) play a key role in host antiviral defense. We have previously screened the porcine 2'-5'-oligoadenylate synthetase-like protein (pOASL) as a potential anti-CSFV ISG using a reporter CSFV. This study aimed to clarify the underlying antiviral mechanism of pOASL against CSFV. We confirmed that CSFV replication was significantly suppressed in lentivirus-delivered, pOASL-overexpressing PK-15 cells, whereas silencing the expression of endogenous pOASL by small interfering RNAs markedly enhanced CSFV growth. In addition, the transcriptional level of pOASL was upregulated both in vitro and in vivo upon CSFV infection. Interestingly, the anti-CSFV effects of pOASL are independent of the canonical RNase L pathway but depend on the activation of the type I IFN response. Glutathione S -transferase pulldown and coimmunoprecipitation assays revealed that pOASL interacts with MDA5, a double-stranded RNA sensor, and further enhances MDA5-mediated type I IFN signaling. Moreover, we showed that pOASL exerts anti-CSFV effects in an MDA5-dependent manner. In conclusion, pOASL suppresses CSFV replication via the MDA5-mediated type I IFN-signaling pathway. IMPORTANCE The host innate immune response plays an important role in mounting the initial resistance to viral infection. Here, we identify the porcine 2'-5'-oligoadenylate synthetase-like protein (pOASL) as an interferon (IFN)-stimulated gene (ISG) against classical swine fever virus (CSFV). We demonstrate that the anti-CSFV effects of pOASL depend on the activation of type I IFN response. In addition, we show that pOASL, as an MDA5-interacting protein, is a coactivator of MDA5-mediated IFN induction to exert anti-CSFV actions. This work will be beneficial to the development of novel anti-CSFV strategies by targeting pOASL. Copyright © 2017 Li et al.

  18. African swine fever virus infection in Classical swine fever subclinically infected wild boars.

    PubMed

    Cabezón, Oscar; Muñoz-González, Sara; Colom-Cadena, Andreu; Pérez-Simó, Marta; Rosell, Rosa; Lavín, Santiago; Marco, Ignasi; Fraile, Lorenzo; de la Riva, Paloma Martínez; Rodríguez, Fernando; Domínguez, Javier; Ganges, Llilianne

    2017-08-01

    Recently moderate-virulence classical swine fever virus (CSFV) strains have been proven capable of generating postnatal persistent infection (PI), defined by the maintenance of viremia and the inability to generate CSFV-specific immune responses in animals. These animals also showed a type I interferon blockade in the absence of clinical signs. In this study, we assessed the infection generated in 7-week-old CSFV PI wild boars after infection with the African swine fever virus (ASFV). The wild boars were divided in two groups and were infected with ASFV. Group A comprised boars who were CSFV PI in a subclinical form and Group B comprised pestivirus-free wild boars. Some relevant parameters related to CSFV replication and the immune response of CSFV PI animals were studied. Additionally, serum soluble factors such as IFN-α, TNF-α, IL-6, IL-10, IFN-γ and sCD163 were analysed before and after ASFV infection to assess their role in disease progression. After ASFV infection, only the CSFV PI wild boars showed progressive acute haemorrhagic disease; however, the survival rates following ASFV infection was similar in both experimental groups. Notwithstanding, the CSFV RNA load of CSFV PI animals remained unaltered over the study; likewise, the ASFV DNA load detected after infection was similar between groups. Interestingly, systemic type I FN-α and IL-10 levels in sera were almost undetectable in CSFV PI animals, yet detectable in Group B, while detectable levels of IFN-γ were found in both groups. Finally, the flow cytometry analysis showed an increase in myelomonocytic cells (CD172a + ) and a decrease in CD4 + T cells in the PBMCs from CSFV PI animals after ASFV infection. Our results showed that the immune response plays a role in the progression of disease in CSFV subclinically infected wild boars after ASFV infection, and the immune response comprised the systemic type I interferon blockade. ASFV does not produce any interference with CSFV replication, or vice versa. ASFV infection could be a trigger factor for the disease progression in CSFV PI animals, as their survival after ASFV was similar to that of the pestivirus-free ASFV-infected group. This fact suggests a high resistance in CSFV PI animals even against a virus like ASFV; this may mean that there are relevant implications for CSF control in endemic countries. The diagnosis of ASFV and CSFV co-infection in endemic countries cannot be ruled out and need to be studied in greater depth.

  19. Effects of the nuclear localization of the N(pro) protein of classical swine fever virus on its virulence in pigs.

    PubMed

    Li, Yongfeng; Shen, Liang; Sun, Yuan; Wang, Xiao; Li, Chao; Huang, Junhua; Chen, Jianing; Li, Lianfeng; Zhao, Bibo; Luo, Yuzi; Li, Su; Qiu, Hua-Ji

    2014-12-05

    The N(pro) protein of classical swine fever virus (CSFV) is localized in the cytoplasm and nucleus. However, it is unknown whether the nuclear localization of N(pro) correlates with the virulence of CSFV in the host. Previously, we showed that the N(pro) protein fused with interferon regulatory factor 3 (IRF3) was present only in the cytoplasm. Here, we generated and evaluated a recombinant CSFV vSM-IRF3 harboring the IRF3 gene inserted into the N(pro) gene of the highly virulent CSFV Shimen strain. Compared to the even nuclear and cytoplasmic distribution of the enhanced green fluorescent protein (EGFP)-N(pro) fusion expressed by the recombinant CSFV EGFP-CSFV, vSM-IRF3 expressed an IRF3-N(pro) fusion protein that only was localized in the cytoplasm. vSM-IRF3 was markedly attenuated in vitro and in vivo, and the inoculated pigs were completely protected from lethal CSFV challenge, whereas the parental virus as well as EGFP-CSFV exhibited a typical virulent phenotype. Taken together, the nuclear localization of N(pro) plays a significant role in the CSFV replication and virulence. Copyright © 2014 Elsevier B.V. All rights reserved.

  20. Classical swine fever virus replicated poorly in cells from MxA transgenic pigs.

    PubMed

    Zhao, Yicheng; Wang, Tiedong; Yao, Li; Liu, Bo; Teng, Chunbo; Ouyang, Hongsheng

    2016-08-17

    In addition to their value as livestock, pigs are susceptible to classical swine fever virus (CSFV) and can serve as reservoirs for CSFV, allowing it to develop into an epizootic. CSFV, a pestivirus of the Flaviviridae family, has a single-stranded RNA genome. Recent research has indicated that the human MxA protein inhibits the life cycles of certain RNA viruses, such as members of the Bunyaviridae family, the Flaviviridae family and others. To produce pigs with antiviral protection against CSFV, transgenic pigs expressing human MxA were generated by nuclear transplantation. Cells from three MxA transgenic piglets were used to investigate in vitro antiviral activity of MxA aganist CSFV, and the results of in vitro indirect immunofluorescence assays, virus titration and real-time PCR indicated that the MxA transgenic pig has an antiviral capacity against CSFV. Transgene with human MxA on pigs is feasible. High levels of MxA expression do inhibit CSFV in vitro at early time points post-infection at 60-96dpi.

  1. Porcine Mx1 Protein Inhibits Classical Swine Fever Virus Replication by Targeting Nonstructural Protein NS5B.

    PubMed

    Zhou, Jing; Chen, Jing; Zhang, Xiao-Min; Gao, Zhi-Can; Liu, Chun-Chun; Zhang, Yun-Na; Hou, Jin-Xiu; Li, Zhao-Yao; Kan, Lin; Li, Wen-Liang; Zhou, Bin

    2018-04-01

    Mx proteins are interferon (IFN)-induced GTPases that have broad antiviral activity against a wide range of RNA and DNA viruses; they belong to the dynamin superfamily of large GTPases. In this study, we confirmed the anti-classical swine fever virus (CSFV) activity of porcine Mx1 in vitro and showed that porcine Mx2 (poMx2), human MxA (huMxA), and mouse Mx1 (mmMx1) also have anti-CSFV activity in vitro Small interfering RNA (siRNA) experiments revealed that depletion of endogenous poMx1 or poMx2 enhanced CSFV replication, suggesting that porcine Mx proteins are responsible for the antiviral activity of interferon alpha (IFN-α) against CSFV infection. Confocal microscopy, immunoprecipitation, glutathione S -transferase (GST) pulldown, and bimolecular fluorescence complementation (BiFC) demonstrated that poMx1 associated with NS5B, the RNA-dependent RNA polymerase (RdRp) of CSFV. We used mutations in the poMx1 protein to elucidate the mechanism of their anti-CSFV activity and found that mutants that disrupted the association with NS5B lost all anti-CSV activity. Moreover, an RdRp activity assay further revealed that poMx1 undermined the RdRp activities of NS5B. Together, these results indicate that porcine Mx proteins exert their antiviral activity against CSFV by interacting with NS5B. IMPORTANCE Our previous studies have shown that porcine Mx1 (poMx1) inhibits classical swine fever virus (CSFV) replication in vitro and in vivo , but the molecular mechanism of action remains largely unknown. In this study, we dissect the molecular mechanism of porcine Mx1 and Mx2 against CSFV in vitro Our results show that poMx1 associates with NS5B, the RNA-dependent RNA polymerase of CSFV, resulting in the reduction of CSFV replication. Moreover, the mutants of poMx1 further elucidate the mechanism of their anti-CSFV activities. Copyright © 2018 American Society for Microbiology.

  2. An investigation of classical swine fever virus seroprevalence and risk factors in pigs in East Nusa Tenggara, eastern Indonesia.

    PubMed

    Sawford, Kate; Geong, Maria; Bulu, Petrus M; Drayton, Emily; Mahardika, Gusti N K; Leslie, Edwina E C; Robertson, Ian; Gde Putra, Anak Agung; Toribio, Jenny-Ann L M L

    2015-05-01

    Classical swine fever virus (CSFV) is a highly infectious disease of pigs. It has had significant impacts on East Nusa Tenggara, eastern Indonesia since its introduction in 1997. In spite of its importance to this region, little is known about its seroprevalence and distribution, and pig-level and farmer-level factors that may have an impact on the serological status of an individual pig. To address this knowledge deficit, a cross-sectional seroprevalence survey was conducted in 2010 involving 2160 pigs and 805 farmers from four islands in the region. Farmer questionnaires and pig record forms were used to collect data about the farmers and pigs surveyed. Blood was collected from each pig to determine its CSFV serological status. Apparent and true prevalence were calculated for each island, district, subdistrict, and village surveyed. CSFV serological status was used as an outcome variable in mixed effects logistic regression analyses. Overall true CSFV seroprevalence was estimated at 17.5% (lower CI 16.0%; upper CI 19.5%). Seroprevalence estimates varied widely across the islands, districts, subdistricts, and villages. Manggarai Barat, a district on the western end of Flores Island, contained pigs that were positive for antibody to CSFV. This result was unexpected, as no clinical cases had been reported in this area. Older pigs and pigs that had been vaccinated for CSFV were more likely to test positive for antibody to CSFV. The final multivariable model accounted for a large amount of variation in the data, however much of this variation was explained by the random effects with less than 2% of the variation explained by pig age and pig CSFV vaccination status. In this study we documented the seroprevalence of CSFV across four islands in East Nusa Tenggara, eastern Indonesia. We also identified risk factors for the presence of antibody to CSFV. Further investigation is needed to understand why clinical CSFV has not been reported on the western end of Flores Island, and to identify additional risk factors that explain CSFV serological status to inform disease control strategies. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. An investigation of classical swine fever virus seroprevalence and risk factors in pigs in Timor-Leste.

    PubMed

    Sawford, Kate; do Karmo, Antonino; da Conceicao, Felisiano; Geong, Maria; Tenaya, I Wayan Masa; Hartawan, Dinar H W; Toribio, Jenny-Ann L M L

    2015-11-01

    Classical swine fever virus (CSFV) is a highly infectious pathogen of pigs and believed to be a major constraint to pig production in Timor-Leste. The Ministry of Agriculture and Fisheries conducts vaccination campaigns in an attempt to control clinical disease, however, there is no empirical data available concerning the seroprevalence and distribution of CSFV in Timor-Leste. To help address this knowledge deficit, a cross-sectional study to determine seroprevalence was conducted in the three districts that border Indonesia. Data on farmer- and pig-level factors were also collected to look at their impact on CSFV serological status. Overall, true CSFV seroprevalence was estimated at 34.4%. Seroprevalence estimates varied widely between and within districts, subdistricts, and villages. Older pigs and pigs that had been vaccinated for CSFV were more likely to test positive for CSFV antibody. Pigs owned by farmers that experienced the sudden death of pigs in the 12 months prior to the survey were more likely to test positive for CSFV antibody, while pigs that had been sick in the previous three months were less likely to test positive for CSFV antibody. The final multivariable model accounted for a large amount of variation in the data, however, much of this variation was explained by the random effects with less than one percent of the variation explained by the fixed effects. This work further supports the need for a collaborative approach to whole-island CSFV control between West Timor, Indonesia and Timor-Leste. Further work is needed to better understand the risk factors for CSFV serological status in order to allocate resources for control. As CSFV is now endemic in Timor-Leste research involving a combination of serology, antigen detection and in-depth investigation of suspect cases over a period of time may be required. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Rab1A is required for assembly of classical swine fever virus particle.

    PubMed

    Lin, Jihui; Wang, Chengbao; Liang, Wulong; Zhang, Jing; Zhang, Longxiang; Lv, Huifang; Dong, Wang; Zhang, Yanming

    2018-01-15

    Rab1A belongs to the small Rab GTPase family and is involved in the lifecycle of numerous viruses. Here, knockdown of Rab1A inhibited CSFV growth. Further study revealed that Rab1A depletion decreased intracellular and extracellular CSFV titers, but did not affect intracellular virus genome copies and E2 protein expression within a virus lifecycle, which suggested that Rab1A is required for CSFV particle assembly rather than for genome replication or virion release. This was proofed by blocking the spread of virus using neutralizing antibodies, through which the negative effects of Rab1A knockdown on multi-cycle replication of CSFV were eliminated. Moreover, co-immunoprecipitation and confocal microscopy assays showed that Rab1A bound to CSFV NS5A protein, indicating that Rab1A and viral NS5A proteins may work cooperatively during CSFV particle assembly. In conclusion, this study demonstrated for the first time that Rab1A is required for CSFV particle assembly and binds to viral particle assembly-related NS5A protein. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. Detection of Classical swine fever virus infection by individual oral fluid of pigs following experimental inoculation.

    PubMed

    Petrini, Stefano; Pierini, Ilaria; Giammarioli, Monica; Feliziani, Francesco; De Mia, Gian Mario

    2017-03-01

    We evaluated the use of oral fluid as an alternative to serum samples for Classical swine fever virus (CSFV) detection. Individual oral fluid and serum samples were collected at different times post-infection from pigs that were experimentally inoculated with CSFV Alfort 187 strain. We found no evidence of CSFV neutralizing antibodies in swine oral fluid samples under our experimental conditions. In contrast, real-time reverse transcription-polymerase chain reaction could detect CSFV nucleic acid from the oral fluid as early as 8 d postinfection, which also coincided with the time of initial detection in blood samples. The probability of CSFV detection in oral fluid was identical or even higher than in the corresponding blood sample. Our results support the feasibility of using this sampling method for CSFV genome detection, which may represent an additional cost-effective tool for CSF control.

  6. Mutation of E1 glycoprotein of classical swine fever virus affects viral virulence in swine.

    PubMed

    Risatti, G R; Holinka, L G; Lu, Z; Kutish, G F; Tulman, E R; French, R A; Sur, J H; Rock, D L; Borca, M V

    2005-12-05

    Transposon linker insertion mutagenesis of a full-length infectious clone (IC) (pBIC) of the pathogenic classical swine fever virus (CSFV) strain Brescia was used to identify genetic determinants of CSFV virulence and host range. Here, we characterize a virus mutant, RB-C22v, possessing a 19-residue insertion at the carboxyl terminus of E1 glycoprotein. Although RB-C22v exhibited normal growth characteristics in primary porcine macrophage cell cultures, the major target cell of CSFV in vivo, it was markedly attenuated in swine. All RB-C22v-infected pigs survived infection remaining clinically normal in contrast to the 100% mortality observed for BICv-infected animals. Comparative pathogenesis studies demonstrated a delay in RB-C22v spread to, and decreased replication in the tonsils, a 10(2) to 10(7) log10 reduction in virus titers in lymphoid tissues and blood, and an overall delay in generalization of infection relative to BICv. Notably, RB-C22v-infected animals were protected from clinical disease when challenged with pathogenic BICv at 3, 5, 7, and 21 days post-RB-C22v inoculation. Viremia, viral replication in tissues, and oronasal shedding were reduced in animals challenged at 7 and 21 DPI. Notably BICv-specific RNA was not detected in tonsils of challenged animals. These results indicate that a carboxyl-terminal domain of E1 glycoprotein affects virulence of CSFV in swine, and they demonstrate that mutation of this domain provides the basis for a rationally designed and efficacious live-attenuated CSF vaccine.

  7. Tissue expression of Toll-like receptors 2, 3, 4 and 7 in swine in response to the Shimen strain of classical swine fever virus

    PubMed Central

    Cao, Zhi; Zheng, Minping; Lv, Huifang; Guo, Kangkang; Zhang, Yanming

    2018-01-01

    The Toll-like receptors (TLRs) of the innate immune system provide the host with the ability to detect and respond to viral infections. The present study aimed to investigate the mRNA and protein expression levels of TLR2, 3, 4 and 7 in porcine tissues upon infection with the highly virulent Shimen strain of classical swine fever virus (CSFV). Reverse transcription-quantitative polymerase chain reaction was used to detect the mRNA expression levels of CSFV and TLR, whereas western blotting was used to detect the expression levels of TLR proteins. In addition, tissues underwent histological examination and immunohistochemistry to reveal the histopathological alterations associated with highly virulent CSFV infection and to detect TLR antigens. Furthermore, porcine monocyte-derived macrophages (pMDMs) were prestimulated with peptidoglycan from Staphylococcus aureus (PGN-SA), polyinosinic-polycytidylic acid [poly (I:C)], lipopolysaccharide from Escherichia coli 055:B5 (LPS-B5) or imiquimod (R837) in order to analyze the association between TLR expression and CSFV replication. Following stimulation for 12 h (with TLR-specific ligands), cells were infected with CSFV Shimen strain. The results revealed that the expression levels of TLR2 and TLR4 were increased in the lung and kidney, but were decreased in the spleen and lymph nodes in response to CSFV. TLR3 was strongly expressed in the heart and slightly upregulated in the spleen in response to CSFV Shimen strain infection, and TLR7 was increased in all examined tissues in the presence of CSFV. Furthermore, R837 and LPS-B5 exerted inhibitory effects on CSFV replication in pMDMs, whereas PGN-SA and poly(I:C) had no significant effect. These findings highlight the potential role of TLR expression in the context of CSFV infection. PMID:29568891

  8. Recoding structural glycoprotein E2 in classical swine fever virus (CSFV) produces complete virus attenuation in swine and protects infected animals against disease

    USDA-ARS?s Scientific Manuscript database

    Controlling classical swine fever (CSF) involves vaccination in endemic regions and preemptive slaughter of infected swine herds during epidemics. Generally, live attenuated vaccines induce solid immunity. Using diverse approaches, reverse genetics has been useful in developing classical swine fever...

  9. Phylodynamics of classical swine fever virus with emphasis on Ecuadorian strains.

    PubMed

    Garrido Haro, A D; Barrera Valle, M; Acosta, A; J Flores, F

    2018-06-01

    Classic swine fever virus (CSFV) is a Pestivirus from the Flaviviridae family that affects pigs worldwide and is endemic in several Latin American countries. However, there are still some countries in the region, including Ecuador, for which CSFV molecular information is lacking. To better understand the epidemiology of CSFV in the Americas, sequences from CSFVs from Ecuador were generated and a phylodynamic analysis of the virus was performed. Sequences for the full-length glycoprotein E2 gene of twenty field isolates were obtained and, along with sequences from strains previously described in the Americas and from the most representative strains worldwide, were used to analyse the phylodynamics of the virus. Bayesian methods were used to test several molecular clock and demographic models. A calibrated ultrametric tree and a Bayesian skyline were constructed, and codons associated with positive selection involving immune scape were detected. The best model according to Bayes factors was the strict molecular clock and Bayesian skyline model, which shows that CSFV has an evolution rate of 3.2 × 10 -4 substitutions per site per year. The model estimates the origin of CSFV in the mid-1500s. There is a strong spatial structure for CSFV in the Americas, indicating that the virus is moving mainly through neighbouring countries. The genetic diversity of CSFV has increased constantly since its appearance, with a slight decrease in mid-twentieth century, which coincides, with eradication campaigns in North America. Even though there is no evidence of strong directional evolution of the E2 gene in CSFV, codons 713, 761, 762 and 975 appear to be selected positively and could be related to virulence or pathogenesis. These results reveal how CSFV has spread and evolved since it first appeared in the Americas and provide important information for attaining the goal of eradication of this virus in Latin America. © 2018 Blackwell Verlag GmbH.

  10. The Laminin Receptor Is a Cellular Attachment Receptor for Classical Swine Fever Virus

    PubMed Central

    Chen, Jianing; He, Wen-Rui; Shen, Liang; Dong, Hong; Yu, Jiahui; Wang, Xiao; Yu, Shaoxiong; Li, Yongfeng; Li, Su; Luo, Yuzi; Sun, Yuan

    2015-01-01

    ABSTRACT Classical swine fever virus (CSFV) is the causative agent of classical swine fever (CSF), a highly contagious, economically important viral disease in many countries. The Erns and E2 envelope glycoproteins are responsible for the binding to and entry into the host cell by CSFV. To date, only one cellular receptor, heparan sulfate (HS), has been identified as being involved in CSFV attachment. HS is also present on the surface of various cells that are nonpermissive to CSFV. Hence, there must be another receptor(s) that has been unidentified to date. In this study, we used a set of small interfering RNAs (siRNAs) against a number of porcine cell membrane protein genes to screen cellular proteins involved in CSFV infection. This approach resulted in the identification of several proteins, and of these, the laminin receptor (LamR) has been demonstrated to be a cellular receptor for several viruses. Confocal analysis showed that LamR is colocalized with CSFV virions on the membrane, and a coimmunoprecipitation assay indicated that LamR interacts with the CSFV Erns protein. In inhibition assays, anti-LamR antibodies, soluble laminin, or LamR protein significantly inhibited CSFV infection in a dose-dependent manner. Transduction of PK-15 cells with a recombinant lentivirus expressing LamR yielded higher viral titers. Moreover, an attachment assay demonstrated that LamR functions during virus attachment. We also demonstrate that LamR acts as an alternative attachment receptor, especially in SK6 cells. These results indicate that LamR is a cellular attachment receptor for CSFV. IMPORTANCE Classical swine fever virus (CSFV) is the causative agent of classical swine fever (CSF), an economically important viral disease affecting the pig industry in many countries. To date, only heparan sulfate (HS) has been identified to be an attachment receptor for CSFV. Here, using RNA interference screening with small interfering RNAs (siRNAs) against a number of porcine membrane protein genes, we identified the laminin receptor (LamR) to be another attachment receptor. We demonstrate the involvement of LamR together with HS in virus attachment, and we elucidate the relationship between LamR and HS. LamR also serves as an attachment receptor for many viral pathogens, including dengue virus, a fatal human flavivirus. The study will help to enhance our understanding of the life cycle of flaviviruses and the development of antiviral strategies for flaviviruses. PMID:25694590

  11. Porcine circovirus type 2 (PCV2) infection decreases the efficacy of an attenuated classical swine fever virus (CSFV) vaccine

    PubMed Central

    2011-01-01

    The Lapinized Philippines Coronel (LPC) vaccine, an attenuated strain of classical swine fever virus (CSFV), is an important tool for the prevention and control of CSFV infection and is widely and routinely used in most CSF endemic areas, including Taiwan. The aim of this study was to investigate whether PCV2 infection affects the efficacy of the LPC vaccine. Eighteen 6-week-old, cesarean-derived and colostrum-deprived (CDCD), crossbred pigs were randomly assigned to four groups. A total of 105.3 TCID50 of PCV2 was experimentally inoculated into pigs through both intranasal and intramuscular routes at 0 days post-inoculation (dpi) followed by LPC vaccination 12 days later. All the animals were challenged with wild-type CSFV (ALD stain) at 27 dpi and euthanized at 45 dpi. Following CSFV challenge, the LPC-vaccinated pigs pre-inoculated with PCV2 showed transient fever, viremia, and viral shedding in the saliva and feces. The number of IgM+, CD4+CD8-CD25+, CD4+CD8+CD25+, and CD4-CD8+CD25+ lymphocyte subsets and the level of neutralizing antibodies against CSFV were significantly higher in the animals with LPC vaccination alone than in the pigs with PCV2 inoculation/LPC vaccination. In addition, PCV2-derived inhibition of the CSFV-specific cell proliferative response of peripheral blood mononuclear cells (PBMCs) was demonstrated in an ex vivo experiment. These findings indicate that PCV2 infection decreases the efficacy of the LPC vaccine. This PCV2-derived interference may not only allow the invasion of wild-type CSFV in pig farms but also increases the difficulty of CSF prevention and control in CSF endemic areas. PMID:22129109

  12. The effect of classical swine fever virus NS5A and NS5A mutants on oxidative stress and inflammatory response in swine testicular cells.

    PubMed

    Dong, Wang; Lv, Huifang; Wang, Yifan; Li, Xiaomeng; Li, Cheng; Wang, Lu; Wang, Chengbao; Guo, Kangkang; Zhang, Yanming

    2017-06-01

    Infection with classical swine fever virus (CSFV) results in highly significant economic losses; this infection is characterized by being highly contagious and accompanied by hyperthermia and systemic bleeding. Oxidative stress (OS) plays a critical role in the pathological process of viral infection. The function of the nonstructural protein 5A (NS5A) in the pathogenesis of CSFV has not been completely understood. Here, OS and the inflammatory response were studied with NS5A and substitution mutants in swine testicular (ST) cells. ST cell lines stably expressing CSFV NS5A or substitution mutants were established. Reactive oxygen species (ROS) production, antioxidant protein expression and inflammatory response were analyzed by quantitative real-time PCR (qRT-PCR), ELISA and flow cytometry analysis. The results showed that CSFV NS5A did not increase ROS production or the antioxidant protein (Trx, HO-1 and PRDX-6) expression in ST cells. However, NS5A inhibited cyclooxygenase-2 (COX-2) expression, a pro-inflammatory protein related to OS. Further studies have shown that NS5A mutants S15A and S92A increased ROS production and inhibited antioxidant protein expression. S15A, S81A and T274A affected the inflammatory response. This study suggested that CSFV NS5A did not induce OS, and amino acids Ser15 and Ser92 of CSFV NS5A were essential for inhibiting OS. Additionally, Ser15, Ser81 and Thr274 played important roles in the inflammatory response in ST cells. These observations provided insight into the function of CSFV NS5A and the mechanism of CSFV persistent infection in ST cells. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Classical swine fever virus induces pyroptosis in the peripheral lymphoid organs of infected pigs.

    PubMed

    Yuan, Jin; Zhu, Mengjiao; Deng, Shaofeng; Fan, Shuangqi; Xu, Hailuan; Liao, Jiedan; Li, Peng; Zheng, Jingfang; Zhao, Mingqiu; Chen, Jinding

    2018-05-02

    Classical swine fever virus (CSFV) causes a highly lethal disease in pigs, which is characterized by immunosuppression. Leukopenia is known to be a possible mechanism of immunosuppression during CSFV infection. As a new and specialized form of cell death, pyroptosis is the key response of the innate immune system to pathogens, and is widely involved in the occurrence and development of infectious diseases. However, the relationship between CSFV and pyroptosis has not been explored. In this study, we investigated the occurrence of pyroptosis in pigs following CSFV infection. According to qRT-PCR assay results, the prevalence of this virus in peripheral lymphoid organs (tonsils, lymph nodes, and spleen) was much higher than that in other organs. Severe bleeding, necrosis, and a significant reduction in lymphocytes were found in the peripheral lymphoid organs of CSFV-infected pigs based on histological examination. In-depth studies showed that an increased ratio of deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-positive cells were present in the peripheral lymphoid organs of the CSFV-infected group according to immunohistochemistry. Meanwhile, the p10 subunit and activity of caspase-1, which is a regulator of pyroptosis, the N-terminal domain of gasdermin D, which is an executor of pyroptosis, and the cleavage and secretion of IL-1b, which is a product of pyroptosis were increased in the peripheral lymphoid organs of the CSFV-infected group. Together, these results demonstrated that pyroptosis is involved in CSFV-induced cell death in vivo, which provides a new understanding of the mechanism associated with lymphocyte depletion and immunosuppression in pigs infected with this virus. Copyright © 2018 Elsevier B.V. All rights reserved.

  14. Specific ligands for classical swine fever virus screened from landscape phage display library.

    PubMed

    Yin, Long; Luo, Yuzi; Liang, Bo; Wang, Fei; Du, Min; Petrenko, Valery A; Qiu, Hua-Ji; Liu, Aihua

    2014-09-01

    Classical swine fever (CSF) is a devastating infectious disease caused by classical swine fever virus (CSFV). The screening of CSFV-specific ligands is of great significance for diagnosis and treatment of CSF. Affinity selection from random peptide libraries is an efficient approach to discover ligands with high stability and specificity. Here, we screened phage ligands for the CSFV E2 protein from f8/8 landscape phage display library by biopanning and obtained four phage clones specific for the E2 protein of CSFV. Viral blocking assays indicated that the phage clone displaying the octapeptide sequence DRATSSNA remarkably inhibited the CSFV replication in PK-15 cells at a titer of 10(10) transduction units, as evidenced by significantly decreased viral RNA copies and viral titers. The phage-displayed E2-binding peptides have the potential to be developed as antivirals for CSF. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. The Npro product of classical swine fever virus and bovine viral diarrhea virus uses a conserved mechanism to target interferon regulatory factor-3.

    PubMed

    Seago, Julian; Hilton, Louise; Reid, Elizabeth; Doceul, Virginie; Jeyatheesan, Janan; Moganeradj, Kartykayan; McCauley, John; Charleston, Bryan; Goodbourn, Stephen

    2007-11-01

    Classical swine fever virus (CSFV) is a member of the genus Pestivirus in the family Flaviviridae. The N(pro) product of CSFV targets the host's innate immune response and can prevent the production of type I interferon (IFN). The mechanism by which CSFV orchestrates this inhibition was investigated and it is shown that, like the related pestivirus bovine viral diarrhea virus (BVDV), this involves the N(pro) protein targeting interferon regulatory factor-3 (IRF-3) for degradation by proteasomes and thus preventing IRF-3 from activating transcription from the IFN-beta promoter. Like BVDV, the steady-state levels of IRF-3 mRNA are not reduced markedly by CSFV infection or N(pro) overexpression. Moreover, IFN-alpha stimulation of CSFV-infected cells induces the antiviral protein MxA, indicating that, as in BVDV-infected cells, the JAK/STAT pathway is not targeted for inhibition.

  16. Detection of classical swine fever virus E2 gene in cattle serum samples from cattle herds of Meghalaya.

    PubMed

    Chakraborty, A K; Karam, A; Mukherjee, P; Barkalita, L; Borah, P; Das, S; Sanjukta, R; Puro, K; Ghatak, S; Shakuntala, I; Sharma, I; Laha, R G; Sen, A

    2018-03-01

    The present study focused on the detection and genetic characterisation of 5' untranslated region (5'UTR) and E2 gene of classical swine fever virus (CSFV, family Flaviviridae , genus Pestivirus ) from bovine population of the northeastern region of India. A total of 134 cattle serum samples were collected from organised cattle farms and were screened for CSFV antigen with a commercial antigen capture enzyme linked immunosorbent assay (Ag-ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). A total of 10 samples were positive for CSFV antigen by ELISA, while all of them were positive in PCR for 5'UTR region. Full length E2 region of CSFV were successfully amplified from two positive samples and used for subsequent phylogenetic analysis and determination of protein 3D structure which showed similarity with reported CSFV isolate from Assam of sub-genogroup 2.1, with minor variations in protein structure.

  17. First molecular identification and characterization of classical swine fever virus isolates from Nepal.

    PubMed

    Postel, Alexander; Jha, Vijay C; Schmeiser, Stefanie; Becher, Paul

    2013-01-01

    Classical swine fever (CSF) is a major constraint to pig production worldwide, and in many developing countries, the epidemiological status is unknown. Here, for the first time, molecular identification and characterization of CSFV isolates from two recent outbreaks in Nepal are presented. Analysis of full-length E2-encoding sequences revealed that these isolates belonged to CSFV subgenotype 2.2 and had highest genetic similarity to isolates from India. Hence, for CSFV, Nepal and India should be regarded as one epidemiological unit. Both Nepalese isolates exhibited significant sequence differences, excluding a direct epidemiological connection and suggesting that CSFV is endemic in that country.

  18. Immunogenicity of Recombinant Classic Swine Fever Virus CD8+ T Lymphocyte Epitope and Porcine Parvovirus VP2 Antigen Coexpressed by Lactobacillus casei in Swine via Oral Vaccination ▿

    PubMed Central

    Xu, Yigang; Cui, Lichun; Tian, Changyong; Zhang, Guocai; Huo, Guicheng; Tang, Lijie; Li, Yijing

    2011-01-01

    Classical swine fever virus (CSFV) and porcine parvovirus (PPV) are highly contagious pathogens, resulting in enormous economic losses in pig industries worldwide. Because vaccines play an important role in disease control, researchers are seeking improved vaccines that could induce antiviral immune responses against CSFV and PPV at the mucosal and systemic levels simultaneously. In this study, a genetically engineered Lactobacillus strain coexpressing the CSFV-specific cytotoxic T lymphocyte (CTL) epitope 290 and the VP2 antigen of PPV was developed, and its immunopotentiating capacity as an oral vaccine in pigs was analyzed. The data demonstrated that in the absence of any adjuvant, the recombinant Lactobacillus strain can efficiently stimulate mucosal and systemic CSFV-specific CD8+ CTL responses to protect pigs against CSFV challenge. Moreover, anti-PPV-VP2 serum IgG and mucosal IgA were induced in pigs immunized orally with the recombinant Lactobacillus strain, showing a neutralizing effect on PPV infection. The results suggest that the recombinant Lactobacillus microecological agent may be a valuable component of a strategy for development of a vaccine against CSFV and PPV. PMID:21940406

  19. Crowd Sourced Formal Verification-Augmentation (CSFV-A)

    DTIC Science & Technology

    2016-06-01

    Formal Verification (CSFV) program built games that recast FV problems into puzzles to make these problems more accessible, increasing the manpower to...construct FV proofs. This effort supported the CSFV program by hosting the games on a public website, and analyzed the gameplay for efficiency to...provide FV proofs. 15. SUBJECT TERMS Crowd Source, Software, Formal Verification, Games 16. SECURITY CLASSIFICATION OF: 17. LIMITATION OF ABSTRACT

  20. Optical high-resolution analysis of rotational movement: testing circular spatial filter velocimetry (CSFV) with rotating biological cells

    NASA Astrophysics Data System (ADS)

    Schaeper, M.; Schmidt, R.; Kostbade, R.; Damaschke, N.; Gimsa, J.

    2016-07-01

    Circular spatial filtering velocimetry (CSFV) was tested during the microscopic registration of the individual rotations of baker’s yeast cells. Their frequency-dependent rotation (electrorotation; ER) was induced in rotating electric fields, which were generated in a glass chip chamber with four electrodes (600 μm tip-to-tip distance). The electrodes were driven with sinusoidal quadrature signals of 5 or 8 V PP with frequencies up to 3 MHz. The observed cell rotation was of the order of 1-100 s per revolution. At each measuring frequency, the independent rotations of up to 20 cells were simultaneously recorded with a high-speed camera. CSFV was software-implemented using circular spatial filters with harmonic gratings. ER was proportional to the phase shift between the values of the spatial filtering signal of consecutive frames. ER spectra obtained by CSFV from the rotation velocities at different ER-field frequencies agreed well with manual measurements and theoretical spectra. Oscillations in the rotation velocity of a single cell in the elliptically polarized field near an electrode, which were resolved by CSFV, could not be visually discerned. ER step responses after field-on were recorded at 2500 frames per second. Analysis proved the high temporal resolution of CSFV and revealed a largely linear torque-friction relation during the acceleration phase of ER. Future applications of CSFV will allow for the simple and cheap automated high-resolution analysis of rotational movements where mechanical detection has too low a resolution or is not possible, e.g. in polluted environments or for gas and fluid vortices, microscopic objects, etc.

  1. Classical Swine Fever in Wild Hog: Report of its Prevalence in Northeast India.

    PubMed

    Barman, N N; Bora, D P; Khatoon, E; Mandal, S; Rakshit, A; Rajbongshi, G; Depner, K; Chakraborty, A; Kumar, S

    2016-10-01

    Classical swine fever virus (CSFV) is the causative agent of a highly contagious disease, hog cholera in pigs. The disease is endemic in many parts of the world and vaccination is the only way to protect the animals from CSFV infection. Wild hogs belong to the species Sus Scrofa Cristatus under the family Suidae are quite susceptible to CSFV infection. The epidemiological role concerning classical swine fever (CSF) in India is largely unknown. We report here the three isolated cases of CSF in wild hogs from three National parks, namely Kaziranga National Park, Manas National Park and Jaldapara National Park, from north-east part of India. The post-mortem and histopathological findings were clearly indicative for CSFV infection. The presence of CSFV genome was demonstrated in several organs and tissues collected from hogs died due to viral infection. In addition, CSF-specific antibodies were detected in two wild hogs as well as in eighteen feral pigs from the same locations. The phylogenetic analysis of the partial E2 protein gene and 5' untranslated region of CSFV isolates from the wild hog showed identities with genotype 2.2 of the Indian isolates. Occurrence of CSF in wild hogs may pose a potent threat in the epidemiology of the virus in Northeast part of India. To the best of our knowledge, the report presented in the manuscript is the first comprehensive report on CSF in wild hogs form Northeast India. The findings reported would help us to understand the epidemiology and biology of CSFV in wild animals. © 2014 Blackwell Verlag GmbH.

  2. Detection of antibodies against classical swine fever virus in fecal samples from wild boar.

    PubMed

    Seo, Sang won; Sunwoo, Sun young; Hyun, Bang hoon; Lyoo, Young S

    2012-12-28

    Classical swine fever (CSF) is a contagious viral disease that affects pigs. Wild boars can play an important epidemiological role in CSF outbreaks. In the past decades, studies conducted in many countries have reported that the CSF virus (CSFV) may persist in wild boar populations. The existence of CSFV in the free-ranging wild boar populations was indirectly confirmed by determining the prevalence of antibodies against CSFV in the serum of hunted wild boars. However, analyzing sero-prevalence in hunted wild boars to study the risk of CSF outbreaks is difficult due to insufficient number of samples, limitation of hunting area and biased age distribution of hunted wild boars. To improve this survey method, we collected feces of wild boars from their habitat and tested them using CSFV antibody enzyme-linked immunosorbent assay (ELISA) and CSF virus neutralization (VN) test. In this study, ELISA was found to be highly sensitive for detecting antibodies against CSFV in fecal samples. Most of doubtful or positive results obtained in CSFV ELISA were confirmed by VN tests. Despite the high coincidence rate of antibody-positive samples between CSFV ELISA and VN test, the possibility of false positive reaction should be considered. In the regional distribution, a fact that antibody-positive fecal and serum samples were found in geographically close area was shown. Hence, presence of antibodies in fecal samples may provide vital information regarding the risk of CSF outbreaks in wild boar groups in geographical proximity. Copyright © 2012 Elsevier B.V. All rights reserved.

  3. Evaluation of an Erns-based enzyme-linked immunosorbent assay to distinguish Classical swine fever virus-infected pigs from pigs vaccinated with CP7_E2alf.

    PubMed

    Pannhorst, Katrin; Fröhlich, Andreas; Staubach, Christoph; Meyer, Denise; Blome, Sandra; Becher, Paul

    2015-07-01

    Infections with Classical swine fever virus (CSFV) are a major economic threat to pig production. To combat CSF outbreaks and to maintain trade, new marker vaccines were developed that allow differentiation of infected from vaccinated animals (DIVA principle). The chimeric pestivirus CP7_E2alf was shown to be safe and efficacious. Its DIVA strategy is based on the detection of CSFV E(rns)-specific antibodies that are only developed on infection. However, for the new marker vaccine to be considered a valuable control tool, a validated discriminatory assay is needed. One promising candidate is the already commercially available enzyme-linked immunosorbent assay, PrioCHECK CSFV E(rns) ELISA (Prionics BV, Lelystad, The Netherlands). Four laboratories of different European Union member states tested 530 serum samples and country-specific field sera from domestic pigs and wild boar. The ELISA displayed a good robustness. However, based on its reproducibility and repeatability, ranges rather than single values for diagnostic sensitivity and specificity were defined. The ELISA displayed a sensitivity of 90-98% with sera from CSFV-infected domestic pigs. A specificity of 89-96% was calculated with sera from domestic pigs vaccinated once with CP7_E2alf. The ELISA detected CSFV infections in vaccinated domestic pigs with a sensitivity of 82-94%. The sensitivity was lower with sera taken ≤21 days post-challenge indicating that the stage of CSFV infection had a considerable influence on testing. Taken together, the PrioCHECK CSFV E(rns) ELISA can be used for detection of CSFV infections in CP7_E2alf-vaccinated and nonvaccinated domestic pig populations, but should only be applied on a herd basis by testing a defined number of animals. © 2015 The Author(s).

  4. A bivalent dendrimeric peptide bearing a T-cell epitope from foot-and-mouth disease virus protein 3A improves humoral response against classical swine fever virus.

    PubMed

    Bohórquez, José Alejandro; Defaus, Sira; Muñoz-González, Sara; Perez-Simó, Marta; Rosell, Rosa; Fraile, Lorenzo; Sobrino, Francisco; Andreu, David; Ganges, Llilianne

    2017-06-15

    Three dendrimeric peptides were synthesized in order to evaluate their immunogenicity and their potential protection against classical swine fever virus (CSFV) in domestic pigs. Construct 1, an optimized version of a previously used dendrimer, had four copies of a B-cell epitope derived from CSFV E2 glycoprotein connected to an also CSFV-derived T-cell epitope through maleimide instead of thioether linkages. Construct 2 was similarly built but included only two copies of the B-cell epitope, and in also bivalent construct 3 the CSFV T-cell epitope was replaced by a previously described one from the 3A protein of foot-and-mouth disease virus (FMDV). Animals were inoculated twice with a 21-day interval and challenged 15days after the second immunization. Clinical signs were recorded daily and ELISA tests were performed to detect antibodies against specific peptide and E2. The neutralising antibody response was assessed 13days after challenge. Despite the change to maleimide connectivity, only partial protection against CSFV was again observed. The best clinical protection was observed in group 3. Animals inoculated with constructs 2 and 3 showed higher anti-peptide humoral response, suggesting that two copies of the B-cell epitope are sufficient or even better than four copies for swine immune recognition. In addition, for construct 3 higher neutralizing antibody titres against CSFV were detected. Our results support the immunogenicity of the CSFV B-cell epitope and the cooperative role of the FMDV 3A T-cell epitope in inducing a neutralising response against CSFV in domestic pigs. This is also the first time that the FMDV T-cell epitope shows effectivity in improving swine immune response against a different virus. Our findings highlight the relevance of dendrimeric peptides as a powerful tool for epitope characterization and antiviral strategies development. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Co-expression of Erns and E2 genes of classical swine fever virus by replication-defective recombinant adenovirus completely protects pigs against virulent challenge with classical swine fever virus.

    PubMed

    Sun, Yongke; Yang, Yuai; Zheng, Huanli; Xi, Dongmei; Lin, Mingxing; Zhang, Xiaomin; Yang, Linfu; Yan, Yulin; Chu, Xiaohui; Bi, Baoliang

    2013-04-01

    The objective of this study was to construct a recombinant adenovirus for future CSFV vaccines used in the pig industry for the reduction of losses involved in CSF outbreaks. The Erns and E2 genes of classical swine fever virus (CSFV), which encode the two main protective glycoproteins from the "Shimen" strain of CSFV, were combined and inserted into the replication-defective human adenovirus type-5 and named the rAd-Erns-E2. Nine pigs were randomly assigned to three treatment groups (three pigs in each group) including the rAd-Erns-E2, hAd-CMV control and DMEM control. Intramuscular vaccination with 2×10(6) TCID(50) of the rAd-Erns-E2 was administered two times with an interval of 21 days. At 42 days post inoculation, pigs in all groups were challenged with a lethal dose of 1×10(3) TCID(50) CSFV "Shimen" strain. Observation of clinical signs was made and the existence of CSFV RNA was detected. Animals in the hAd-CMV and DMEM groups showed severe clinical CSF symptoms and were euthanized from 7 to 10 days after the challenge. However, no adverse clinical CSF signs were observed in vaccinated pigs after the administration of rAd-Erns-E2 and even after CSFV challenge. Neither CSFV RNA nor pathological changes were detected in the tissues of interest of the above vaccinated pigs. These results implied that the recombination adenovirus carrying the Erns-E2 genes could be used to prevent swine from classical swine fever. Copyright © 2012 Elsevier Ltd. All rights reserved.

  6. Absence of autophagy promotes apoptosis by modulating the ROS-dependent RLR signaling pathway in classical swine fever virus-infected cells

    PubMed Central

    Pei, Jingjing; Deng, Jieru; Ye, Zuodong; Wang, Jiaying; Gou, Hongchao; Liu, Wenjun; Zhao, Mingqiu; Liao, Ming; Yi, Lin; Chen, Jinding

    2016-01-01

    ABSTRACT A growing number of studies have demonstrated that both macroautophagy/autophagy and apoptosis are important inner mechanisms of cell to maintain homeostasis and participate in the host response to pathogens. We have previously reported that a functional autophagy pathway is trigged by infection of classical swine fever virus (CSFV) and is required for viral replication and release in host cells. However, the interplay of autophagy and apoptosis in CSFV-infected cells has not been clarified. In the present study, we demonstrated that autophagy induction with rapamycin facilitates cellular proliferation after CSFV infection, and that autophagy inhibition by knockdown of essential autophagic proteins BECN1/Beclin 1 or MAP1LC3/LC3 (microtubule-associated protein 1 light chain 3) promotes apoptosis via fully activating both intrinsic and extrinsic mechanisms in CSFV-infected cells. We also found that RIG-I-like receptor (RLR) signaling was amplified in autophagy-deficient cells during CSFV infection, which was closely linked to the activation of the intrinsic apoptosis pathway. Moreover, we discovered that virus infection of autophagy-impaired cells results in an increase in copy number of mitochondrial DNA and in the production of reactive oxygen species (ROS), which plays a significant role in enhanced RLR signaling and the activated extrinsic apoptosis pathway in cultured cells. Collectively, these data indicate that CSFV-induced autophagy delays apoptosis by downregulating ROS-dependent RLR signaling and thus contributes to virus persistent infection in host cells. PMID:27463126

  7. Absence of autophagy promotes apoptosis by modulating the ROS-dependent RLR signaling pathway in classical swine fever virus-infected cells.

    PubMed

    Pei, Jingjing; Deng, Jieru; Ye, Zuodong; Wang, Jiaying; Gou, Hongchao; Liu, Wenjun; Zhao, Mingqiu; Liao, Ming; Yi, Lin; Chen, Jinding

    2016-10-02

    A growing number of studies have demonstrated that both macroautophagy/autophagy and apoptosis are important inner mechanisms of cell to maintain homeostasis and participate in the host response to pathogens. We have previously reported that a functional autophagy pathway is trigged by infection of classical swine fever virus (CSFV) and is required for viral replication and release in host cells. However, the interplay of autophagy and apoptosis in CSFV-infected cells has not been clarified. In the present study, we demonstrated that autophagy induction with rapamycin facilitates cellular proliferation after CSFV infection, and that autophagy inhibition by knockdown of essential autophagic proteins BECN1/Beclin 1 or MAP1LC3/LC3 (microtubule-associated protein 1 light chain 3) promotes apoptosis via fully activating both intrinsic and extrinsic mechanisms in CSFV-infected cells. We also found that RIG-I-like receptor (RLR) signaling was amplified in autophagy-deficient cells during CSFV infection, which was closely linked to the activation of the intrinsic apoptosis pathway. Moreover, we discovered that virus infection of autophagy-impaired cells results in an increase in copy number of mitochondrial DNA and in the production of reactive oxygen species (ROS), which plays a significant role in enhanced RLR signaling and the activated extrinsic apoptosis pathway in cultured cells. Collectively, these data indicate that CSFV-induced autophagy delays apoptosis by downregulating ROS-dependent RLR signaling and thus contributes to virus persistent infection in host cells.

  8. A Promising Trigene Recombinant Human Adenovirus Vaccine Against Classical Swine Fever Virus.

    PubMed

    Li, Helin; Gao, Rui; Zhang, Yanming

    2016-05-01

    Classical swine fever (CSF) vaccine based on HAdV-5 had achieved an efficient protection in swine. Both classical swine fever virus (CSFV) E0 glycoprotein and E2 glycoprotein were the targets for neutralizing antibodies and related to immune protection against CSF. Interleukin-2 (IL2), as an adjuvant, also had been used in CSF vaccine research. In this study, coexpression of the CSFV E0, E2, and IL2 genes by HAdV-5 (rAdV-E0-E2-IL2) was constructed and immunized to evaluate its efficacy. Three expressed genes had been sequentially connected with foot-and-mouth disease virus 2A (FMDV 2A). The vaccine was administered by intramuscular inoculation to CSFV-free pigs (10(8) TCID50) twice at triweekly intervals. No adverse clinical signs were observed in any of the pigs after vaccination. The vaccine induced strong humoral and cellular responses that led to complete protection against clinical signs of lethal CSFV infection, viremia, and shedding of challenge virus. The rAdV-E0-E2-IL2 is a promising, efficient, and safe marker vaccine candidate against CSFV.

  9. Genetic diversity of subgenotype 2.1 isolates of classical swine fever virus.

    PubMed

    Gong, Wenjie; Wu, Jianmin; Lu, Zongji; Zhang, Li; Qin, Shaomin; Chen, Fenglian; Peng, Zhicheng; Wang, Qin; Ma, Ling; Bai, Anbin; Guo, Huancheng; Shi, Jishu; Tu, Changchun

    2016-07-01

    As the causative agent of classical swine fever, the economically devastating swine disease worldwide, classical swine fever virus (CSFV) is currently classified into the 11 subgenotypes, of which subgenotype 2.1 is distributed worldwide and showing more genetic diversity than other subgenotypes. Prior to this report, subgenotype 2.1 was divided into three sub-subgenotypes (2.1a-2.1c). To further analyze the genetic diversity of CSFV isolates in China, 39 CSFV isolates collected between 2004 and 2012 in two Chinese provinces Guangxi and Guangdong were sequenced and subjected to phylogenetic analysis together with reference sequences retrieved from GenBank. Phylogenetic analyses based on the 190-nt and/or 1119-nt full length E2 gene fragments showed that current CSFV subgenotype 2.1 virus isolates in the world could be divided into 10 sub-subgenotypes (2.1a-2.1j) and the 39 isolates collected in this study were grouped into 7 of them (2.1a-2.1c and 2.1g-2.1j). Among the 10 sub-subgenotypes, 2.1d-2.1j were newly identified. Sub-subgenotype 2.1d isolates were circulated only in India, however the rest 9 sub-subgenotypes were from China with some of them closely related to isolates from European and neighboring Asian countries. According to the temporal and spatial distribution of CSFV subgenotype 2.1 isolates, the newly classified 10 sub-subgenotypes were further categorized into three groups: dominant sub-subgenotype, minor sub-subgenotype and silent sub-subgenotype, and each sub-subgenotype can be found only in certain geographical areas. Taken together, this study reveals the complex genetic diversity of CSFV subgenotype 2.1 and improves our understanding about the epidemiological trends of CSFV subgenotype 2.1 in the world, particularly in China. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Selection of Classical Swine Fever Virus with Enhanced Pathogenicity Reveals Synergistic Virulence Determinants in E2 and NS4B

    PubMed Central

    Tamura, Tomokazu; Yoshino, Fumi; Nomura, Takushi; Yamamoto, Naoki; Sato, Yuka; Okamatsu, Masatoshi; Ruggli, Nicolas; Kida, Hiroshi

    2012-01-01

    Classical swine fever virus (CSFV) is the causative agent of classical swine fever (CSF), a highly contagious disease of pigs. There are numerous CSFV strains that differ in virulence, resulting in clinical disease with different degrees of severity. Low-virulent and moderately virulent isolates cause a mild and often chronic disease, while highly virulent isolates cause an acute and mostly lethal hemorrhagic fever. The live attenuated vaccine strain GPE− was produced by multiple passages of the virulent ALD strain in cells of swine, bovine, and guinea pig origin. With the aim of identifying the determinants responsible for the attenuation, the GPE− vaccine virus was readapted to pigs by serial passages of infected tonsil homogenates until prolonged viremia and typical signs of CSF were observed. The GPE−/P-11 virus isolated from the tonsils after the 11th passage in vivo had acquired 3 amino acid substitutions in E2 (T830A) and NS4B (V2475A and A2563V) compared with the virus before passages. Experimental infection of pigs with the mutants reconstructed by reverse genetics confirmed that these amino acid substitutions were responsible for the acquisition of pathogenicity. Studies in vitro indicated that the substitution in E2 influenced virus spreading and that the changes in NS4B enhanced the viral RNA replication. In conclusion, the present study identified residues in E2 and NS4B of CSFV that can act synergistically to influence virus replication efficiency in vitro and pathogenicity in pigs. PMID:22674973

  11. Numerical simulation of the wave-induced non-linear bending moment of ships

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Xia, J.; Wang, Z.; Gu, X.

    1995-12-31

    Ships traveling in moderate or rough seas may experience non-linear bending moments due to flare effect and slamming loads. The numerical simulation of the total wave-induced bending moment contributed from both the wave frequency component induced by wave forces and the high frequency whipping component induced by slamming actions is very important in predicting the responses and ensuring the safety of the ship in rough seas. The time simulation is also useful for the reliability analysis of ship girder strength. The present paper discusses four different methods of the numerical simulation of wave-induced non-linear vertical bending moment of ships recentlymore » developed in CSSRC, including the hydroelastic integral-differential method (HID), the hydroelastic differential analysis method (HDA), the combined seakeeping and structural forced vibration method (CSFV), and the modified CSFV method (MCSFV). Numerical predictions are compared with the experimental results obtained from the elastic ship model test of S-175 container ship in regular and irregular waves presented by Watanabe Ueno and Sawada (1989).« less

  12. Comprehensive analysis of the codon usage patterns in the envelope glycoprotein E2 gene of the classical swine fever virus

    PubMed Central

    Chi, Xiaojuan; Wang, Song; Ma, Yanmei; Chen, Jilong

    2017-01-01

    The classical swine fever virus (CSFV), circulating worldwide, is a highly contagious virus. Since the emergence of CSFV, it has caused great economic loss in swine industry. The envelope glycoprotein E2 gene of the CSFV is an immunoprotective antigen that induces the immune system to produce neutralizing antibodies. Therefore, it is essential to study the codon usage of the E2 gene of the CSFV. In this study, 140 coding sequences of the E2 gene were analyzed. The value of effective number of codons (ENC) showed low codon usage bias in the E2 gene. Our study showed that codon usage could be described mainly by mutation pressure ENC plot analysis combined with principal component analysis (PCA) and translational selection-correlation analysis between the general average hydropathicity (Gravy) and aromaticity (Aroma), and nucleotides at the third position of codons (A3s, T3s, G3s, C3s and GC3s). Furthermore, the neutrality analysis, which explained the relationship between GC12s and GC3s, revealed that natural selection had a key role compared with mutational bias during the evolution of the E2 gene. These results lay a foundation for further research on the molecular evolution of CSFV. PMID:28880881

  13. Comprehensive analysis of the codon usage patterns in the envelope glycoprotein E2 gene of the classical swine fever virus.

    PubMed

    Chen, Ye; Li, Xinxin; Chi, Xiaojuan; Wang, Song; Ma, Yanmei; Chen, Jilong

    2017-01-01

    The classical swine fever virus (CSFV), circulating worldwide, is a highly contagious virus. Since the emergence of CSFV, it has caused great economic loss in swine industry. The envelope glycoprotein E2 gene of the CSFV is an immunoprotective antigen that induces the immune system to produce neutralizing antibodies. Therefore, it is essential to study the codon usage of the E2 gene of the CSFV. In this study, 140 coding sequences of the E2 gene were analyzed. The value of effective number of codons (ENC) showed low codon usage bias in the E2 gene. Our study showed that codon usage could be described mainly by mutation pressure ENC plot analysis combined with principal component analysis (PCA) and translational selection-correlation analysis between the general average hydropathicity (Gravy) and aromaticity (Aroma), and nucleotides at the third position of codons (A3s, T3s, G3s, C3s and GC3s). Furthermore, the neutrality analysis, which explained the relationship between GC12s and GC3s, revealed that natural selection had a key role compared with mutational bias during the evolution of the E2 gene. These results lay a foundation for further research on the molecular evolution of CSFV.

  14. Propagation of classical swine fever virus in vitro circumventing heparan sulfate-adaptation.

    PubMed

    Eymann-Häni, Rita; Leifer, Immanuel; McCullough, Kenneth C; Summerfield, Artur; Ruggli, Nicolas

    2011-09-01

    Amplification of natural virus isolates in permanent cell lines can result in adaptation, in particular enhanced binding to heparan sulfate (HS)-containing glycosaminoglycans present on most vertebrate cells. This has been reported for several viruses, including the pestivirus classical swine fever virus (CSFV), the causative agent of a highly contagious hemorrhagic disease in pigs. Propagation of CSFV in cell culture is essential in virus diagnostics and research. Adaptation of CSFV to HS-binding has been related to amino acid changes in the viral E(rns) glycoprotein, resulting in viruses with altered replication characteristics in vitro and in vivo. Consequently, a compound blocking the HS-containing structures on cell surfaces was employed to monitor conversion from HS-independency to HS-dependency. It was shown that the porcine PEDSV.15 cell line permitted propagation of CSFV within a limited number of passages without adaptation to HS-binding. The selection of HS-dependent CSFV mutants was also prevented by propagation of the virus in the presence of DSTP 27. The importance of these findings can be seen from the altered ratio of cell-associated to secreted virus upon acquisition of enhanced HS-binding affinity, a phenotype proposed previously to be related to virulence in the natural host. Copyright © 2011 Elsevier B.V. All rights reserved.

  15. Experimental infection of Bama miniature pigs with a highly virulent classical swine fever virus.

    PubMed

    Sun, Yuan; Jiang, Qian; Tian, Da-Yong; Lin, Huan; Li, Hong; Han, Qiu-Ying; Han, Wen; Si, Chang-De; Hu, Shou-Ping; Zhang, Zhuo; Qu, Lian-Dong; Qiu, Hua-Ji

    2011-09-25

    Currently, larger domestic pigs are only animals widely used in vaccine evaluation and pathogenicity study of classical swine fever virus (CSFV). This study was aimed to create an alternative animal experimental infection model of CSFV. Twenty specific-pathogen-free Bama miniature pigs were randomly divided into two groups and rooms, infected and non-infected, and the pigs in the infected group were inoculated intramuscularly with 104, 105 or 106 TCID50 (median tissue culture infective dose) CSFV Shimen strain (n = 5 × 3) or left uninoculated to serve as in-contact pigs (n = 3). The uninfected control pigs (n = 2) were housed in a separate room. Clinical signs, body temperature, viraemia, tissue antigen distribution, pathological changes and seroconversion were monitored. Clinical signs were observed as early as 2 days post-inoculation (dpi) in all infected pigs (though mild in contact pigs), but not non-infected control pigs. All inoculated pigs showed viraemia by 6 dpi. The in-contact pigs showed lower levels of viraemia. At 10 dpi, seroconversion was noted in five of the 15 inoculated pigs. All inoculated or one in-contact pigs died by 15 dpi. These results show that Bama miniature pigs support productive CSFV infection and display clinical signs and pathological changes consistent with CSFV infections observed in larger domestic pigs.

  16. Secreted Expression of the Cap Gene of Porcine Circovirus Type 2 in Classical Swine Fever Virus C-Strain: Potential of C-Strain Used as a Vaccine Vector

    PubMed Central

    Zhang, Lingkai; Li, Yongfeng; Xie, Libao; Wang, Xiao; Gao, Xulei; Sun, Yuan; Qiu, Hua-Ji

    2017-01-01

    Bivalent vaccines based on live attenuated viruses expressing a heterologous protein are an attractive strategy to address co-infections with various pathogens in the field. Considering the excellent efficacy and safety of the lapinized live attenuated vaccine C-strain (HCLV strain) of classical swine fever virus (CSFV), we proposed that C-strain has the potential as a viral vector for developing bivalent vaccines. To this end, we generated three recombinant viruses based on C-strain, one expressing the capsid (Cap) gene of porcine circovirus type 2 (PCV2) with the nuclear localization signal (NLS) (rHCLV-2ACap), and the other two expressing the PCV2 Cap gene without the NLS yet containing the signal peptide of the prolactin gene (rHCLV-pspCap) or that of the ubiquitin-specific peptidase gene (rHCLV-uspCap). All the recombinant viruses exhibited phenotypes similar to those of the parental virus and produced high-level anti-CSFV neutralizing antibodies (NAbs) in rabbits. Interestingly, rHCLV-uspCap and rHCLV-pspCap, but not rHCLV-2ACap, elicited detectable anti-Cap and -PCV2 NAbs in rabbits. Taken together, our data demonstrate that C-strain can be used as a viral vector to develop bivalent vaccines. PMID:29035292

  17. Secreted Expression of the Cap Gene of Porcine Circovirus Type 2 in Classical Swine Fever Virus C-Strain: Potential of C-Strain Used as a Vaccine Vector.

    PubMed

    Zhang, Lingkai; Li, Yongfeng; Xie, Libao; Wang, Xiao; Gao, Xulei; Sun, Yuan; Qiu, Hua-Ji

    2017-10-16

    Bivalent vaccines based on live attenuated viruses expressing a heterologous protein are an attractive strategy to address co-infections with various pathogens in the field. Considering the excellent efficacy and safety of the lapinized live attenuated vaccine C-strain (HCLV strain) of classical swine fever virus (CSFV), we proposed that C-strain has the potential as a viral vector for developing bivalent vaccines. To this end, we generated three recombinant viruses based on C-strain, one expressing the capsid ( Cap ) gene of porcine circovirus type 2 (PCV2) with the nuclear localization signal (NLS) (rHCLV-2ACap), and the other two expressing the PCV2 Cap gene without the NLS yet containing the signal peptide of the prolactin gene (rHCLV-pspCap) or that of the ubiquitin-specific peptidase gene (rHCLV-uspCap). All the recombinant viruses exhibited phenotypes similar to those of the parental virus and produced high-level anti-CSFV neutralizing antibodies (NAbs) in rabbits. Interestingly, rHCLV-uspCap and rHCLV-pspCap, but not rHCLV-2ACap, elicited detectable anti-Cap and -PCV2 NAbs in rabbits. Taken together, our data demonstrate that C-strain can be used as a viral vector to develop bivalent vaccines.

  18. Isolation and Characterization of a Moderately Virulent Classical Swine Fever Virus Emerging in China.

    PubMed

    Luo, Y; Ji, S; Liu, Y; Lei, J-L; Xia, S-L; Wang, Y; Du, M-L; Shao, L; Meng, X-Y; Zhou, M; Sun, Y; Qiu, H-J

    2017-12-01

    Classical swine fever (CSF) is a devastating infectious disease of pigs caused by classical swine fever virus (CSFV). In China, CSF has been under control owing to extensive vaccination with the lapinized attenuated vaccine (C-strain) since 1950s, despite sporadic or endemic in many regions. However, recently, CSF outbreaks occurred in a large number of swine herds in China. Here, we isolated 15 CSFV strains from diverse C-strain-vaccinated pig farms in China and characterized the genetic variations and antigenicity of the new isolates. The new strains showed unique variations in the E2 protein and were clustered to the subgenotype 2.1d of CSFV recently emerging in China in the phylogenetic tree. Cross-neutralization test showed that the neutralizing titres of porcine anti-C-strain sera against the new isolates were substantially lower than those against both the highly virulent Shimen strain and the subgenotype 2.1b strains that were isolated in China in 2006 and 2009, respectively. In addition, experimental animal infection showed that the HLJZZ2014 strain-infected pigs displayed lower mortality and less severe clinical signs and pathological changes compared with the Shimen strain-infected pigs. The HLJZZ2014 strain was defined to be moderately virulent based on a previously established assessment system for CSFV virulence evaluation, and the virus shedding and the viral load in various tissues of the CSFV HLJZZ2014 strain-infected pigs were significantly lower than those of the Shimen strain-infected pigs. Taken together, the subgenotype 2.1d isolate of CSFV is a moderately virulent strain with molecular variations and antigenic alterations. © 2016 Blackwell Verlag GmbH.

  19. Recombinant Swinepox Virus Expressing Glycoprotein E2 of Classical Swine Fever Virus Confers Complete Protection in Pigs upon Viral Challenge.

    PubMed

    Lin, Huixing; Ma, Zhe; Chen, Lei; Fan, Hongjie

    2017-01-01

    Classical swine fever (CSF) is a highly contagious and serious viral disease that affects the pig industry worldwide. The glycoprotein E2 of the classical swine fever virus (CSFV) can induce neutralizing antibodies, and it is widely used for novel vaccine development. To explore the development of a vaccine against CSFV infections, the gene of glycoprotein E2 was inserted into the swinepox virus (SPV) genome by homologous recombination. The culture titers of rSPV-E2 remained at about 4.3 × 10 6 TCID 50 for more than 60 passages in PK15 and swine testis cell lines. The rSPV-E2 could not be replicated in Vero, MDBK or other non-porcine cell lines. After two to three passages, the SPV specific gene of rSPV-E2 could not been detected in the non-porcine cell culture. To evaluate the immunogenicity of rSPV-E2, 20 CSFV seronegative minipigs were immunized with rSPV-E2, a commercial C-strain vaccine, wild-type SPV (wtSPV; negative control), or PBS (a no-challenge control). After challenge with CSFV, pigs in the rSPV-E2-immunized group showed significantly shorter fever duration compared with the wtSPV-treated group ( P  < 0.05). E2-specific antibodies in the rSPV-E2-immunized group increased dramatically after vaccination and increased continuously over time. CSFV genomic copies in the serum of rSPV-E2-immunized pigs were significantly less compared with the wtSPV-treated group at all time points after challenge ( P  < 0.01). Significant reduction in gross lung lesion scores, histopathological liver, spleen, lung, and kidney lesion scores were noted in the rSPV-E2-immunized group compared with the wtSPV-treated group ( P  < 0.01). The results suggested that the recombinant rSPV-E2 provided pigs with significant protection from CSFV infections; thus, rSPV-E2 offers proof of principle for the development of a vaccine for the prevention of CSFV infections in pigs.

  20. Classical Swine Fever Virus vs. Classical Swine Fever Virus: The Superinfection Exclusion Phenomenon in Experimentally Infected Wild Boar.

    PubMed

    Muñoz-González, Sara; Pérez-Simó, Marta; Colom-Cadena, Andreu; Cabezón, Oscar; Bohórquez, José Alejandro; Rosell, Rosa; Pérez, Lester Josué; Marco, Ignasi; Lavín, Santiago; Domingo, Mariano; Ganges, Llilianne

    2016-01-01

    Two groups with three wild boars each were used: Group A (animals 1 to 3) served as the control, and Group B (animals 4 to 6) was postnatally persistently infected with the Cat01 strain of CSFV (primary virus). The animals, six weeks old and clinically healthy, were inoculated with the virulent strain Margarita (secondary virus). For exclusive detection of the Margarita strain, a specific qRT-PCR assay was designed, which proved not to have cross-reactivity with the Cat01 strain. The wild boars persistently infected with CSFV were protected from superinfection by the virulent CSFV Margarita strain, as evidenced by the absence of clinical signs and the absence of Margarita RNA detection in serum, swabs and tissue samples. Additionally, in PBMCs, a well-known target for CSFV viral replication, only the primary infecting virus RNA (Cat01 strain) could be detected, even after the isolation in ST cells, demonstrating SIE at the tissue level in vivo. Furthermore, the data analysis of the Margarita qRT-PCR, by means of calculated ΔCt values, supported that PBMCs from persistently infected animals were substantially protected from superinfection after in vitro inoculation with the Margarita virus strain, while this virus was able to infect naive PBMCs efficiently. In parallel, IFN-α values were undetectable in the sera from animals in Group B after inoculation with the CSFV Margarita strain. Furthermore, these animals were unable to elicit adaptive humoral (no E2-specific or neutralising antibodies) or cellular immune responses (in terms of IFN-γ-producing cells) after inoculation with the second virus. Finally, a sequence analysis could not detect CSFV Margarita RNA in the samples tested from Group B. Our results suggested that the SIE phenomenon might be involved in the evolution and phylogeny of the virus, as well as in CSFV control by vaccination. To the best of our knowledge, this study was one of the first showing efficient suppression of superinfection in animals, especially in the absence of IFN-α, which might be associated with the lack of innate immune mechanisms.

  1. Efficacy evaluation of the C-strain-based vaccines against the subgenotype 2.1d classical swine fever virus emerging in China.

    PubMed

    Luo, Yuzi; Ji, Shengwei; Lei, Jian-Lin; Xiang, Guang-Tao; Liu, Yan; Gao, Yao; Meng, Xing-Yu; Zheng, Guanglai; Zhang, En-Yu; Wang, Yimin; Du, Ming-Liang; Li, Yongfeng; Li, Su; He, Xi-Jun; Sun, Yuan; Qiu, Hua-Ji

    2017-03-01

    Classical swine fever (CSF) is a devastating infectious disease of pigs caused by classical swine fever virus (CSFV). The disease has been controlled following extensive vaccination with the lapinized attenuated vaccine C-strain for decades in China. However, frequent CSF outbreaks occurred recently in a large number of C-strain-vaccinated pig farms in China and a new subgenotype 2.1d of CSFV has been reported to be responsible for the outbreaks. Here we analyzed the molecular variations and antigenic differences among the C-strain-based commercial vaccines of different origins from different manufacturers in China, and reevaluated the vaccines against the emerging subgenotype 2.1d strain of CSFV. The results showed that the C-strain adapted to the continuous ST cell line (C ST ) contain a unique M290K variation on the E2 protein, compared to those of primary BT cells (C BT ) or rabbit origin (C RT ) and the traditional C-strain sequences available in the GenBank database. Serum neutralization test revealed the antigenic differences between C ST and C BT or C RT . Notably, the neutralizing titers of porcine anti-C-strain sera against the CSFV isolate of subgenotype 2.1d were significantly lower than those against C-strain or Shimen strain. The C-strain-vaccinated, subgenotype 2.1d HLJZZ2014 strain-challenged pigs did not show any clinical signs and all survived. However, these pigs displayed mild pathological and histological lesions, and the CSFV viral RNA was detected in the various tissue and blood samples. Taken together, the C-strain-based vaccines of different origins showed molecular variations and antigenic differences, and could provide clinical but not pathological and virological protection against the subgenotype 2.1d CSFV emerging in China. Further investigation is needed to comprehensively assess the efficacy of C-strain of different doses against the subgenotype 2.1d CSFV. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Sequence adaptations during growth of rescued classical swine fever viruses in cell culture and within infected pigs.

    PubMed

    Hadsbjerg, Johanne; Friis, Martin B; Fahnøe, Ulrik; Nielsen, Jens; Belsham, Graham J; Rasmussen, Thomas Bruun

    2016-08-30

    Classical swine fever virus (CSFV) causes an economically important disease of swine. Four different viruses were rescued from full-length cloned cDNAs derived from the Paderborn strain of CSFV. Three of these viruses had been modified by mutagenesis (with 7 or 8 nt changes) within stem 2 of the subdomain IIIf of the internal ribosome entry site (IRES) that directs the initiation of protein synthesis. Rescued viruses were inoculated into pigs. The rescued vPader10 virus, without modifications in the IRES, induced clinical disease in pigs that was very similar to that observed previously with the parental field strain and transmission to in-contact pigs occurred. Two sequence reversions, in the NS2 and NS5B coding regions, became dominant within the virus populations in these infected pigs. Rescued viruses, with mutant IRES elements, did not induce disease and only very limited circulation of viral RNA could be detected. However, the animals inoculated with these mutant viruses seroconverted against CSFV. Thus, these mutant viruses were highly attenuated in vivo. All 4 rescued viruses were also passaged up to 20 times in cell culture. Using full genome sequencing, the same two adaptations within each of four independent virus populations were observed that restored the coding sequence to that of the parental field strain. These adaptations occurred with different kinetics. The combination of reverse genetics and in depth, full genome sequencing provides a powerful approach to analyse virus adaptation and to identify key determinants of viral replication efficiency in cells and within host animals. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Interaction of CSFV E2 Protein with Swine Host Factors as Detected by Yeast Two-Hybrid System

    PubMed Central

    Gladue, Douglas P.; Baker-Bransetter, Ryan; Holinka, Lauren G.; Fernandez-Sainz, Ignacio J.; O’Donnell, Vivian; Fletcher, Paige; Lu, Zhiqiang; Borca, Manuel V.

    2014-01-01

    E2 is one of the envelope glycoproteins of pestiviruses, including classical swine fever virus (CSFV) and bovine viral diarrhea virus (BVDV). E2 is involved in several critical functions, including virus entry into target cells, induction of a protective immune response and virulence in swine. However, there is no information regarding any host binding partners for the E2 proteins. Here, we utilized the yeast two-hybrid system and identified fifty-seven host proteins as positive binding partners which bound E2 from both CSFV and BVDV with the exception of two proteins that were found to be positive for binding only to CSFV E2. Alanine scanning of CSFV E2 demonstrated that the binding sites for these cellular proteins on E2 are likely non-linear binding sites. The possible roles of the identified host proteins are discussed as the results presented here will be important for future studies to elucidate mechanisms of host protein-virus interactions during pestivirus infection. However, due to the limitations of the yeast two hybrid system, the proteins identified is not exhaustive and each interaction identified needs to be confirmed by independent experimental approaches in the context of virus-infected cells before any definitive conclusion can be drawn on relevance for the virus life cycle. PMID:24416391

  4. Post-Natal Persistent Infection With Classical Swine Fever Virus in Wild Boar: A Strategy for Viral Maintenance?

    PubMed

    Cabezón, O; Colom-Cadena, A; Muñoz-González, S; Pérez-Simó, M; Bohórquez, J A; Rosell, R; Marco, I; Domingo, M; Lavín, S; Ganges, L

    2017-04-01

    In this study, fifteen wild boar piglets were intranasally inoculated <10 h after birth with the moderately virulent classical swine fever virus (CSFV) strain Catalonia 01. At 5 days post-inoculation, seven other animals within 48 h of birth were put in contact with them. Viral replication and innate and specific immune responses were evaluated. Of the inoculated animals, 46.67% remained post-natally persistently infected and were apparently healthy with neither humoral nor cellular immunological responses specific to CSFV and with high viral loads in their blood, organs and body secretions. Moreover, the present data extend the time period to 48 h after birth when a moderately virulent CSFV strain could lead to post-natal persistent infection given the generation of persistently infected wild boars in the contact group (33.33%). The innate immune response to the virus, as measured by type I IFN-α in serum, was mostly not impaired in the persistently infected wild boars. Interestingly, a decrease and lack of IFN-γ-producing cells against CSFV and PHA was observed. In endemic countries where wild swine species are increasing and low and moderate virulence CSFV strains are prevalent, the possible generation of this form of disease cannot be ruled out. © 2015 Blackwell Verlag GmbH.

  5. Recovery of infectious classical swine fever virus (CSFV) from full-length genomic cDNA clones by a swine kidney cell line expressing bacteriophage T7 RNA polymerase.

    PubMed

    van Gennip, H G; van Rijn, P A; Widjojoatmodjo, M N; Moormann, R J

    1999-03-01

    A new method for the recovery of infectious classical swine fever virus (CSFV) from full-length genomic cDNA clones of the C-strain was developed. Classical reverse genetics is based on transfection of in vitro transcribed RNA to target cells to recover RNA viruses. However, the specific infectivity of such in vitro transcribed RNA in swine kidney cells is usually low. To improve reverse genetics for CSFV, a stable swine kidney cell line was established that expresses cytoplasmic bacteriophage T7 RNA polymerase (SK6.T7). A 200-fold increased virus titre was obtained from SK6.T7 cells transfected with linearized full-length cDNA compared to in vitro transcribed RNA, whereas transfection of circular full-length cDNA resulted in 20-fold increased virus titres. Viruses generated on the SK6.T7 cells are indistinguishable from the viruses generated by the classical reverse genetic procedures. These results show the improved recovery of infectious CSFV directly from full-length cDNAs. Furthermore, the reverse genetic procedures are simplified to a faster, one step protocol. We conclude that the SK6.T7 cell line will be a valuable tool for recovering mutant CSFV and will contribute to future pestivirus research.

  6. Effects of glycosylation on antigenicity and immunogenicity of classical swine fever virus envelope proteins

    USDA-ARS?s Scientific Manuscript database

    Classical swine fever virus (CSFV) harbors three envelope glycoproteins (E(rns), E1 and E2). Previous studies have demonstrated that removal of specific glycosylation sites within these proteins yielded attenuated and immunogenic CSFV mutants. Here we analyzed the effects of lack of glycosylation of...

  7. Npro of classical swine fever virus contributes to pathogenicity in pigs by preventing type I interferon induction at local replication sites.

    PubMed

    Tamura, Tomokazu; Nagashima, Naofumi; Ruggli, Nicolas; Summerfield, Artur; Kida, Hiroshi; Sakoda, Yoshihiro

    2014-04-17

    Classical swine fever (CSF) caused by CSF virus (CSFV) is a highly contagious disease of pigs. The viral protein Npro of CSFV interferes with alpha- and beta-interferon (IFN-α/β) induction by promoting the degradation of interferon regulatory factor 3 (IRF3). During the establishment of the live attenuated CSF vaccine strain GPE-, Npro acquired a mutation that abolished its capacity to bind and degrade IRF3, rendering it unable to prevent IFN-α/β induction. In a previous study, we showed that the GPE- vaccine virus became pathogenic after forced serial passages in pigs, which was attributed to the amino acid substitutions T830A in the viral proteins E2 and V2475A and A2563V in NS4B. Interestingly, during the re-adaptation of the GPE- vaccine virus in pigs, the IRF3-degrading function of Npro was not recovered. Therefore, we examined whether restoring the ability of Npro to block IFN-α/β induction of both the avirulent and moderately virulent GPE--derived virus would enhance pathogenicity in pigs. Viruses carrying the N136D substitution in Npro regained the ability to degrade IRF3 and suppress IFN-α/β induction in vitro. In pigs, functional Npro significantly reduced the local IFN-α mRNA expression in lymphoid organs while it increased quantities of IFN-α/β in the circulation, and enhanced pathogenicity of the moderately virulent virus. In conclusion, the present study demonstrates that functional Npro influences the innate immune response at local sites of virus replication in pigs and contributes to pathogenicity of CSFV in synergy with viral replication.

  8. Treatment with interferon-alpha delays disease in swine infected with a highly virulent CSFV strain

    USDA-ARS?s Scientific Manuscript database

    Classical swine fever (CSF) is an economically significant, highly contagious swine disease. The etiological agent, CSF virus (CSFV), is an enveloped virus with a positive-sense, single-stranded RNA genome, classified as a member of the genus Pestivirus within the family Flaviviridae (Becher et al.,...

  9. Classical swine fever virus infection modulates serum levels of INF-α, IL-8 and TNF-α in 6-month-old pigs.

    PubMed

    von Rosen, T; Lohse, L; Nielsen, J; Uttenthal, Å

    2013-12-01

    Several studies have highlighted the important role of cytokines in disease development of classical swine fever virus (CSFV) infection. In the present study, we examined the kinetics of 7 porcine cytokines in serum from pigs infected with 3 different CSFV strains. Based on the clinical picture in 6-month-old Danish pigs, the strains used for inoculation were classified as being of low (Bergen), low to moderate (Eystrup) and moderate to high (Lithuania) virulence. The cytokines interferon-alpha (INF-α), interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNF-α) showed increased levels after CSFV infection with more or less comparable course in the 3 groups. However, the cytokine level peaked with a 2-3 days delay in pigs infected with the low virulent strain compared to those infected with a moderately or highly virulent strain. These findings may indicate that INF-α, IL-8 and TNF-α are involved in the immune response during CSFV infection with strains of different virulence. Copyright © 2013 Elsevier Ltd. All rights reserved.

  10. Cellular Hsp27 interacts with classical swine fever virus NS5A protein and negatively regulates viral replication by the NF-κB signaling pathway.

    PubMed

    Ling, Shifeng; Luo, Mingyang; Jiang, Shengnan; Liu, Jiayu; Ding, Chunying; Zhang, Qinghuan; Guo, Huancheng; Gong, Wenjie; Tu, Changchun; Sun, Jinfu

    2018-05-01

    Classical swine fever virus (CSFV) nonstructural protein NS5A is a multifunctional protein functioning in regulation of viral genome replication, protein translation and assembly by interaction with viral or host proteins. Here, heat shock protein 27 (Hsp27) has been identified as a novel binding partner of NS5A by using His tag "pull down" coupled with shotgun LC-MS/MS, with interaction of both proteins further confirmed by co-immunoprecipitation and laser confocal assays. In PK-15 cells, silencing of Hsp27 expression by siRNA enhanced CSFV replication, and upregulation of Hsp27 inhibited viral proliferation. Additionally, we have shown that overexpression of Hsp27 increased NF-κB signaling induced by TNFα. Blocking NF-κB signaling in PK-15 cells overexpressing Hsp27 by ammonium pyrrolidinedithiocarbamate (PDTC) eliminated the inhibition of CSFV replication by Hsp27. These findings clearly demonstrate that the inhibition of CSFV replication by Hsp27 is mediated via the NF-κB signaling pathway. Copyright © 2018 Elsevier Inc. All rights reserved.

  11. Effective surveillance for early classical swine fever virus detection will utilize both virus and antibody detection capabilities.

    PubMed

    Panyasing, Yaowalak; Kedkovid, Roongtham; Thanawongnuwech, Roongroje; Kittawornrat, Apisit; Ji, Ju; Giménez-Lirola, Luis; Zimmerman, Jeffrey

    2018-03-01

    Early recognition and rapid elimination of infected animals is key to controlling incursions of classical swine fever virus (CSFV). In this study, the diagnostic characteristics of 10 CSFV assays were evaluated using individual serum (n = 601) and/or oral fluid (n = 1417) samples collected from -14 to 28 days post inoculation (DPI). Serum samples were assayed by virus isolation (VI), 2 commercial antigen-capture enzyme-linked immunosorbent assays (ELISA), virus neutralization (VN), and 3 antibody ELISAs. Both serum and oral fluid samples were tested with 3 commercial real-time reverse transcription-polymerase chain reaction (rRT-PCR) assays. One or more serum samples was positive by VI from DPIs 3 to 21 and by antigen-capture ELISAs from DPIs 6 to 17. VN-positive serum samples were observed at DPIs ≥ 7 and by antibody ELISAs at DPIs ≥ 10. CSFV RNA was detected in serum samples from DPIs 2 to 28 and in oral fluid samples from DPIs 4 to 28. Significant differences in assay performance were detected, but most importantly, no single combination of sample and assay was able to dependably identify CSFV-inoculated pigs throughout the 4-week course of the study. The results show that effective surveillance for CSFV, especially low virulence strains, will require the use of PCR-based assays for the detection of early infections (<14 days) and antibody-based assays, thereafter. Copyright © 2018 Elsevier B.V. All rights reserved.

  12. Restoration of glycoprotein Erns dimerization via pseudoreversion partially restores virulence of classical swine fever virus.

    PubMed

    Tucakov, Anna Katharina; Yavuz, Sabine; Schürmann, Eva-Maria; Mischler, Manjula; Klingebeil, Anne; Meyers, Gregor

    2018-01-01

    The classical swine fever virus (CSFV) represents one of the most important pathogens of swine. The CSFV glycoprotein E rns is an essential structural protein and an important virulence factor. The latter is dependent on the RNase activity of this envelope protein and, most likely, its secretion from the infected cell. A further important feature with regard to its function as a virulence factor is the formation of disulfide-linked E rns homodimers that are found in virus-infected cells and virions. Mutant CSFV lacking cysteine (Cys) 171, the residue responsible for intermolecular disulfide bond formation, were found to be attenuated in pigs (Tews BA, Schürmann EM, Meyers G. J Virol 2009;83:4823-4834). In the course of an animal experiment with such a dimerization-negative CSFV mutant, viruses were reisolated from pigs that contained a mutation of serine (Ser) 209 to Cys. This mutation restored the ability to form disulphide-linked E rns homodimers. In transient expression studies E rns mutants carrying the S209C change were found to form homodimers with about wt efficiency. Also the secretion level of the mutated proteins was equivalent to that of wt E rns . Virus mutants containing the Cys171Ser/Ser209Cys configuration exhibited wt growth rates and increased virulence when compared with the Cys171Ser mutant. These results provide further support for the connection between CSFV virulence and E rns dimerization.

  13. Interaction of CSFV E2 protein with swine host factors as detected by yeast two-hybrid system

    USDA-ARS?s Scientific Manuscript database

    E2 is one of the envelope glycoproteins of pestiviruses, including classical swine fever virus (CSFV) and bovine viral diarrhea virus (BVDV). E2 is involved in several critical functions, including virus entry into target cells, induction of a protective immune response and virulence in swine. Howev...

  14. Interaction between core protein of classical swine fever virus with cellular IQGAP1 proetin appears essential for virulence in swine

    USDA-ARS?s Scientific Manuscript database

    Here we show that IQGAP1, a cellular protein that plays a pivotal role as a regulator of the cytoskeleton affecting cell adhesion, polarization and migration, interacts with Classical Swine Fever Virus (CSFV) Core protein. Sequence analyses identified a defined set of residues within CSFV Core prote...

  15. Identification of an NTPase motif in classical swine fever virus NS4B protein

    USDA-ARS?s Scientific Manuscript database

    Classical swine fever (CSF) is a highly contagious and often fatal disease of swine caused by CSF virus (CSFV), a positive sense single-stranded RNA virus in the genus Pestivirus of the Flaviviridae family. Here, we have identified, within CSFV non-structural (NS) protein NS4B, conserved sequence el...

  16. Development of an optimized protocol for the detection of classical swine fever virus in formalin-fixed, paraffin-embedded tissues by seminested reverse transcription-polymerase chain reaction and comparison with in situ hybridization.

    PubMed

    Ha, S-K; Choi, C; Chae, C

    2004-10-01

    An optimized protocol was developed for the detection of classical swine fever virus (CSFV) in formalin-fixed, paraffin-embedded tissues obtained from experimentally and naturally infected pigs by seminested reverse transcription-polymerase chain reaction (RT-PCR). The results for seminested RT-PCR were compared with those determined by in situ hybridization. The results obtained show that the use of deparaffinization with xylene, digestion with proteinase K, extraction with Trizol LS, followed by seminested RT-PCR is a reliable detection method. An increase in sensitivity was observed as amplicon size decreased. The highest sensitivity for RT-PCR on formalin-fixed, paraffin-embedded tissues RNA was obtained with amplicon sizes less than approximately 200 base pairs. An hybridization signal for CSFV was detected in lymph nodes from 12 experimentally and 12 naturally infected pigs. The sensitivity of seminested RT-PCR compared with in situ hybridization was 100% for CSFV. When only formalin-fixed tissues are available, seminested RT-PCR and in situ hybridization would be useful diagnostic methods for the detection of CSFV nucleic acid.

  17. The eukaryotic translation initiation factor 3 subunit E binds to classical swine fever virus NS5A and facilitates viral replication.

    PubMed

    Liu, Xiaofeng; Wang, Xiaoyu; Wang, Qian; Luo, Mingyang; Guo, Huancheng; Gong, Wenjie; Tu, Changchun; Sun, Jinfu

    2018-02-01

    Classical swine fever virus (CSFV) NS5A protein is a multifunctional protein, playing critical roles in viral RNA replication, translation and assembly. To further explore its functions in viral replication, interaction of NS5A with host factors was assayed using a his-tag "pull down" assay coupled with shotgun LC-MS/MS. Host protein translation initiation factor 3 subunit E was identified as a binding partner of NS5A, and confirmed by co-immunoprecipitation and co-localization analysis. Overexpression of eIF3E markedly enhanced CSFV genomic replication, viral protein expression and production of progeny virus, and downregulation of eIF3E by siRNA significantly decreased viral proliferation in PK-15 cells. Luciferase reporter assay showed an enhancement of translational activity of the internal ribosome entry site of CSFV by eIF3E and a decrease in cellular translation by NS5A. These data indicate that eIF3E plays an important role in CSFV replication, thereby identifying it as a potential target for inhibition of the virus. Copyright © 2017 Elsevier Inc. All rights reserved.

  18. Novel rope-based sampling of classical swine fever shedding in a group of wild boar showing low contagiosity upon experimental infection with a classical swine fever field strain of genotype 2.3.

    PubMed

    Mouchantat, Susan; Globig, Anja; Böhle, Wolfgang; Petrov, Anja; Strebelow, Heinz-Günther; Mettenleiter, Thomas C; Depner, Klaus

    2014-06-04

    Several classical swine fever (CSF) epidemics in wild boar and domestic pigs in Europe during the last decades have been caused by CSF virus (CSFV) strains of genotype 2.3. This genotype is known to be virulent leading to high morbidity and mortality. We experimentally infected two eight months old wild boar with 10(5,5) TCID50 of CSFV genotype 2.3 and kept the animals together with five noninoculated wild boar of the same age. Our original purpose was to evaluate a non-invasive sampling method based on saliva collection using "rope-in-a-bait" sampling baits. While expecting high morbidity, high level of virus shedding and some mortality, we actually observed a subclinical course of infection with an unexpected low contagiosity. The two inoculated animals infected only three contact animals while two contact animals remained uninfected. These findings substantially add to our epidemiological understanding of CSFV circulation in wild boar populations. CSFV infected animals older than six months and in good condition may not shed sufficient virus to transmit infection to all seronegative in-contact animals. The contagiosity in relation to the animal's age is discussed. This supports the hypothesis of silent perpetuation of CSFV in wild boar populations for several months if the wild boar density is sufficiently high. The feasibility of the "rope-in-a-bait" sampling method could be proven during the short viraemic phase of infected animals during the second week of infection. Copyright © 2014 Elsevier B.V. All rights reserved.

  19. CP7_E2alf oral vaccination confers partial protection against early classical swine fever virus challenge and interferes with pathogeny-related cytokine responses

    PubMed Central

    2013-01-01

    The conventional C-strain vaccine induces early protection against classical swine fever (CSF), but infected animals cannot be distinguished from vaccinated animals. The CP7_E2alf marker vaccine, a pestivirus chimera, could be a suitable substitute for C-strain vaccine to control CSF outbreaks. In this study, single oral applications of CP7_E2alf and C-strain vaccines were compared for their efficacy to induce protection against a CSF virus (CSFV) challenge with the moderately virulent Bas-Rhin isolate, in pigs as early as two days post-immunization. This work emphasizes the powerful potential of CP7_E2alf vaccine administered orally by a rapid onset of partial protection similar to that induced by the C-strain vaccine. Furthermore, our results revealed that both vaccinations attenuated the effects induced by CSFV on production of the pig major acute phase protein (PigMAP), IFN-α, IL-12, IL-10, and TGF-β1 cytokines. By this interference, several cytokines that may play a role in the pathogeny induced by moderately virulent CSFV strains were revealed. New hypotheses concerning the role of each of these cytokines in CSFV pathogeny are discussed. Our results also show that oral vaccination with either vaccine (CP7_E2alf or C-strain) enhanced CSFV–specific IgG2 production, compared to infection alone. Interestingly, despite the similar antibody profiles displayed by both vaccines post-challenge, the production of CSFV-specific IgG1 and neutralizing antibodies without challenge was lower with CP7_E2alf vaccination than with C-strain vaccination, suggesting a slight difference in the balance of adaptive immune responses between these vaccines. PMID:23398967

  20. Complex Virus-Host Interactions Involved in the Regulation of Classical Swine Fever Virus Replication: A Minireview.

    PubMed

    Li, Su; Wang, Jinghan; Yang, Qian; Naveed Anwar, Muhammad; Yu, Shaoxiong; Qiu, Hua-Ji

    2017-07-05

    Classical swine fever (CSF), caused by classical swine fever virus (CSFV), is one of the most devastating epizootic diseases of pigs in many countries. Viruses are small intracellular parasites and thus rely on the cellular factors for replication. Fundamental aspects of CSFV-host interactions have been well described, such as factors contributing to viral attachment, modulation of genomic replication and translation, antagonism of innate immunity, and inhibition of cell apoptosis. However, those host factors that participate in the viral entry, assembly, and release largely remain to be elucidated. In this review, we summarize recent progress in the virus-host interactions involved in the life cycle of CSFV and analyze the potential mechanisms of viral entry, assembly, and release. We conclude with future perspectives and highlight areas that require further understanding.

  1. Chimeric classical swine fever (CSF)-Japanese encephalitis (JE) viral particles as a non-transmissible bivalent marker vaccine candidate against CSF and JE infections

    USDA-ARS?s Scientific Manuscript database

    A trans-complemented CSF- JE chimeric viral replicon was constructed using an infectious cDNA clone of the CSF virus (CSFV) Alfort/187 strain. The E2 gene of CSFV Alfort/187 strain was deleted and the resultant plasmid pA187delE2 was inserted by a fragment containing the region coding for a truncate...

  2. Seroprevalence of selected viral pathogens in pigs reared in organized farms of Meghalaya from 2014 to 16

    PubMed Central

    Mukherjee, Priyanka; Karam, Amarjit; Singh, Uttam; Chakraborty, Amit Kumar; Huidrom, Surmani; Sen, Arnab; Sharma, Indu

    2018-01-01

    Aim: A pilot study was carried out to find out the seroprevalence of Porcine circovirus 2 (PCV2), classical swine fever virus (CSFV), and Porcine respiratory and reproductive syndrome virus (PRRS) in pig population of Meghalaya. Materials and Methods: Serum samples were collected from piglets of 40–45 days age group, growers, and sows reared under organized and unorganized management in 11 districts of Meghalaya situated in the Khasi, Jaintia, and Garo hills divisions in the time period of 2014-2016 from apparently healthy and suspected pigs. Seroprevalence of PCV2, CSFV, and PRRS specific antibodies was detected by enzyme-linked immunosorbent assay (ELISA). Results: A total of 1899 serum samples were collected and screened using antibody ELISA kits specific for PCV2, CSFV, and PRRS. The highest antibody prevalence during the selected time periods was detected for PCV2 (80.8% in 2014, 79.1% in 2015, and 96.2% in 2016) followed by CSFV (76.4% in 2014, 66.09% in 2015, and 25.5% in 2016) and PRRS (2.8% in 2014, 2.7% in 2015, and 3.62% in 2016). The result indicates high seroprevalence for PCV2, which can be considered as an inducement factor due to the immunosuppressive nature of the virus, for animals being susceptible to other pathogens in farms where airborne transmission of PCV2 and postweaning multisystemic wasting syndrome among animals reared in close pens can be a major possibility. Conclusions: The data from this study indicates ubiquitous prevalence of PCV2 antibodies in the farm animals along with the endemic presence of swine fever and emergence of PRRS in an organized farm. There are few reports regarding PCV2 infections/outbreaks in pigs associated with reproductive failure from northern and southern part of India, but till date, there are no reports regarding concomitant infection of CSFV and PCV2 from India. Considerable high seropositivity of PCV2 indicates the need for high impact hygiene practice in farms, routine seromonitoring and implementation the vaccination program. To the author’s best knowledge, this is the first documented report on the seroprevalence of PCV2, CSFV, and PRRS from pig population of Meghalaya. PMID:29479156

  3. Toward the development of a one-dose classical swine fever subunit vaccine: antigen titration, immunity onset, and duration of immunity

    PubMed Central

    Madera, Rachel F.; Wang, Lihua; Gong, Wenjie; Burakova, Yulia; Buist, Sterling; Nietfeld, Jerome; Henningson, Jamie; Cino-Ozuna, Ada G.; Tu, Changchun

    2018-01-01

    Highly contagious classical swine fever (CSF) remains a major trade and health problem in the pig industry, resulting in large economic losses worldwide. In CSF-endemic countries, attenuated CSF virus (CSFV) vaccines have been routinely used to control the disease. However, eradication of CSFV in a geographical area would require permanent reduction to zero presence of the virus. It is therefore of paramount importance to develop a safe, potent, and non-infectious CSF vaccine. We have previously reported on a cost-effective CSF E2 subunit vaccine, KNB-E2, which can protect against CSF symptoms in a single dose containing 75 µg of recombinant CSFV glycoprotein E2. In this study, we report on a series of animal studies undertaken to elucidate further the efficacy of KNB-E2. We found that pigs vaccinated with a single KNB-E2 dose containing 25 µg of recombinant CSFV glycoprotein E2 were protected from clinical symptoms of CSF. In addition, KNB-E2-mediated reduction of CSF symptoms was observed at two weeks post-vaccination and the vaccinated pigs continued to exhibit reduced CSF clinical signs when virus challenged at two months and four months post-vaccination. These results suggest that KNB-E2 effectively reduces CSF clinical signs, indicating the potential of this vaccine for safely minimizing CSF-related losses. PMID:29510474

  4. Identification of two internal signal peptide sequences: critical for classical swine fever virus non-structural protein 2 to trans-localize to the endoplasmic reticulum.

    PubMed

    Guo, Kang-kang; Tang, Qing-hai; Zhang, Yan-ming; Kang, Kai; He, Lei

    2011-05-18

    The membrane topology and molecular mechanisms for endoplasmic reticulum (ER) localization of classical swine fever virus (CSFV) non-structural 2 (NS2) protien is unclear. We attempted to elucidate the subcellular localization, and the molecular mechanisms responsible for the localization of this protein in our study. The NS2 gene was amplified by reverse transcription polymerase chain reaction, with the transmembrane region and hydrophilicity of the NS2 protein was predicted by bioinformatics analysis. Twelve cDNAs of the NS2 gene were amplified by the PCR deletion method and cloned into a eukaryotic expression vector, which was transfected into a swine umbilical vein endothelial cell line (SUVEC). Subcellular localization of the NS2 protein was characterized by confocal microscopy, and western blots were carried out to analyze protein expression. Our results showed that the -NH2 terminal of the CSFV NS2 protein was highly hydrophobic and the protein localized in the ER. At least four transmembrane regions and two internal signal peptide sequences (amino acids103-138 and 220-262) were identified and thought to be critical for its trans-localization to the ER. This is the first study to identify the internal signal peptide sequences of the CSFV NS2 protein and its subcellular localization, providing the foundation for further exploration of this protein's function of this protein and its role in CSFV pathogenesis.

  5. Emergence of 2.1. subgenotype of classical swine fever virus in pig population of India in 2011.

    PubMed

    Rajkhowa, T K; Hauhnar, Lalthapui; Lalrohlua, Isaac; Mohanarao G, Jagan

    2014-01-01

    Limited studies are available on molecular epidemiology of classical swine fever virus (CSFV) in India and are restricted to domestic pigs. These studies show the presence of 1.1. genotype. The aim of the present study was to subgenotype four CSFV isolates, two each from the outbreaks of CSF in wild (Sus scrofa) and domestic pigs of Mizoram state, India, in 2011. CSFV isolates were subjected to nucleotide sequencing in E2 and NS5B genomic regions. Phylogenetic analysis of the isolates in both genomic regions was carried out with 39 Indian isolates (4 isolates from the present study of Mizoram state and 35 isolates from the other states of India) and 57 reference sequences retrieved from the GenBank database. Two of the 39 isolates from India were collected from wild boar and were subgenotyped as 2.1. Out of 37 isolates from domestic pigs, only two were subgenotyped as 2.1. The analysis revealed the emergence of 2.1. subgenotype of CSFV in both wild and domestic pigs in India. The isolates from domestic pigs of Mizoram state (CSF/MZ/KOL/73 and CSF/MZ/AIZ/115) were grouped in genotype 1 and subgenotype 1.1., thus confirming that the source of CSF outbreaks in domesticated pigs in Mizoram was not from wild pigs. The current study forms an essential step for better understanding of the epidemiology of 2.1 subgroup as well as the movement and spread of the disease in India.

  6. Humoral and cellular immune response in mice induced by the classical swine fever virus E2 protein fused to the porcine CD154 antigen.

    PubMed

    Sordo, Yusmel; Suárez, Marisela; Caraballo, Rosalina; Sardina, Talía; Brown, Emma; Duarte, Carlos; Lugo, Joanna; Gil, Lázaro; Perez, Danny; Oliva, Ayme; Vargas, Milagros; Santana, Elaine; Valdés, Rodolfo; Rodríguez, María Pilar

    2018-03-01

    The development of subunit vaccines against classical swine fever is a desirable goal, because it allows discrimination between vaccinated and infected animals. In this study, humoral and cellular immune response elicited in inbred BALB/c mice by immunization with a recombinant classical swine fever virus (CSFV) E2 protein fused to porcine CD154 antigen (E2CD154) was assessed. This model was used as a predictor of immune response in swine. Mice were immunized with E2CD154 emulsified in Montanide ISA50V2 or dissolved in saline on days 1 and 21. Another group received E2His antigen, without CD154, in the same adjuvant. Montanide ISA50V2 or saline served as negative controls for each experimental group. Animals immunized with 12.5 and 2.5 μg/dose of E2CD154 developed the highest titers (>1:2000) of CSFV neutralizing antibodies. Moreover, CSFV specific splenocyte gamma-interferon production, measured after seven and twenty-eight days of immunization, was significantly higher in mice immunized with 12.5 μg of E2CD154. As a conclusion, E2CD154 emulsified in Montanide ISA50 V2 was able to induce a potent humoral and an early cellular immune response in inbred BALB/c mice. Therefore, this immunogen might be an appropriate candidate to elicit immune response in swine, control CSF disease and to eliminate CSFV in swine. Copyright © 2018 International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.

  7. Serum Metabolomic Profiling of Piglets Infected with Virulent Classical Swine Fever Virus

    PubMed Central

    Gong, Wenjie; Jia, Junjie; Zhang, Bikai; Mi, Shijiang; Zhang, Li; Xie, Xiaoming; Guo, Huancheng; Shi, Jishu; Tu, Changchun

    2017-01-01

    Classical swine fever (CSF) is a highly contagious swine infectious disease and causes significant economic losses for the pig industry worldwide. The objective of this study was to determine whether small molecule metabolites contribute to the pathogenesis of CSF. Birefly, serum metabolomics of CSFV Shimen strain-infected piglets were analyzed by ultraperformance liquid chromatography/electrospray ionization time-of-flight mass spectrometry (UPLC/ESI-Q-TOF/MS) in combination with multivariate statistical analysis. In CSFV-infected piglets at days 3 and 7 post-infection changes were found in metabolites associated with several key metabolic pathways, including tryptophan catabolism and the kynurenine pathway, phenylalanine metabolism, fatty acid and lipid metabolism, the tricarboxylic acid and urea cycles, branched-chain amino acid metabolism, and nucleotide metabolism. Several pathways involved in energy metabolism including fatty acid biosynthesis and β-oxidation, branched-chain amino acid metabolism, and the tricarboxylic acid cycle were significantly inhibited. Changes were also observed in several metabolites exclusively associated with gut microbiota. The metabolomic profiles indicate that CSFV-host gut microbiome interactions play a role in the development of CSF. PMID:28496435

  8. Virus load in pigs affected with different clinical forms of classical swine fever.

    PubMed

    Rout, M; Saikumar, G

    2012-04-01

    Classical swine fever (CSF) is an endemic disease in India, but the real magnitude of the problem is not known as only outbreaks of acute CSF are reported and many cases of chronic and clinically inapparent forms of the disease, which manifest a confusing clinical picture, remain undiagnosed. The real status of classical swine fever virus (CSFV) infection can only be known by testing pigs with highly specific and sensitive diagnostic assays. To obtain the baseline prevalence of CSFV infection among pigs in an endemic region where no vaccination was being performed, a real-time PCR assay was used to detect viral genetic material in tissue samples collected from a slaughterhouse in the northern state of Uttar Pradesh in India. In total, 1120 slaughtered pigs were examined for the presence of CSF suggestive pathological lesions and tissues from suspected cases were tested for the presence of CSFV antigen and nucleic acids by indirect immuno-peroxidase test and real-time PCR, respectively. Based on the detection of viral genetic material in the tonsils, the prevalence of CSFV infection among slaughtered pigs was found to be 7.67%. Pigs detected positive for viral genome by quantitative real-time PCR assay when categorized into different forms of CSF, depending upon the pathological lesions observed, the viral load in the tonsils of some of the pigs with chronic or clinically inapparent form of the disease was similar to that detected in pigs with acute CSF. The results of the study suggested that the risk posed by pigs with chronic disease or those infected but showing no clinical disease may be relatively higher as they can transmit the virus to new susceptible hosts over a longer period of time. © 2011 Blackwell Verlag GmbH.

  9. Enhanced protective immunity of the chimeric vector-based vaccine rAdV-SFV-E2 against classical swine fever in pigs by a Salmonella bacterial ghost adjuvant.

    PubMed

    Xia, Shui-Li; Lei, Jian-Lin; Du, Mingliang; Wang, Yimin; Cong, Xin; Xiang, Guang-Tao; Li, Lian-Feng; Yu, Shenye; Du, Enqi; Liu, Siguo; Sun, Yuan; Qiu, Hua-Ji

    2016-06-14

    Classical swine fever (CSF) is a highly contagious swine disease caused by classical swine fever virus (CSFV). Previously, we demonstrated that rAdV-SFV-E2, an adenovirus-delivered, Semliki Forest virus replicon-vectored marker vaccine against CSF, is able to protect pigs against lethal CSFV challenge. From an economical point of view, it will be beneficial to reduce the minimum effective dose of the vaccine. This study was designed to test the adjuvant effects of Salmonella enteritidis-derived bacterial ghosts (BG) to enhance the protective immunity of rAdV-SFV-E2 in pigs. Groups of 5-week-old pigs (n = 4) were immunized intramuscularly twice with 10(5) median tissue culture infective doses (TCID50) rAdV-SFV-E2 combined with 10(10) colony forming units (CFU) BG, 10(6) or 10(5) TCID50 rAdV-SFV-E2 alone or 10(10) CFU BG alone at an interval of 3 weeks, and challenged with the highly virulent CSFV Shimen strain at 1 week post-booster immunization. The results show that the pigs inoculated with 10(5) TCID50 rAdV-SFV-E2 plus BG or 10(6) TCID50 rAdV-SFV-E2 alone were completely protected from lethal CSFV challenge, in contrast with the pigs vaccinated with 10(5) TCID50 rAdV-SFV-E2 or BG alone, which displayed partial or no protection following virulent challenge. The data indicate that BG are a promising adjuvant to enhance the efficacy of rAdV-SFV-E2 and possibly other vaccines.

  10. Pathway analysis in blood cells of pigs infected with classical swine fever virus: comparison of pigs that develop a chronic form of infection or recover.

    PubMed

    Hulst, Marcel; Loeffen, Willie; Weesendorp, Eefke

    2013-02-01

    Infection of pigs with CSFV can lead to either acute disease, resulting in death or recovery, or chronic disease. The mechanisms by which CSFV manipulates the pig's first line of defence to establish a chronic infection are poorly understood. Therefore, pigs were infected with moderately virulent CSFV, and whole blood was collected on a regular basis during a period of 18 days. Using whole-genome microarrays, time-dependent changes in gene expression were recorded in blood cells of chronically diseased pigs and pigs that recovered. Bioinformatics analysis of regulated genes indicated that different immunological pathways were regulated in chronically diseased pigs compared to recovered pigs. In recovered pigs, antiviral defence mechanisms were rapidly activated, whereas in chronically diseased pigs, several genes with the potential to inhibit NF-κB- and IRF3/7-mediated transcription of type I interferons were up-regulated. Compared to recovered pigs, chronically diseased pigs failed to activate NK or cytotoxic T-cell pathways, and they showed decreased gene activity in antigen-presenting monocytes/macrophages. Remarkably, in chronically diseased pigs, genes related to the human autoimmune disease systemic lupus erythematosus (SLE) were up-regulated during the whole period of 18 days. CSFV pathology in kidney and skin resembles that of SLE. Furthermore, enzymes involved in the degradation of 1,25-dihydroxyvitamin D3 and of tryptophan to kynurenines were expressed at different levels in chronically diseased and recovered pigs. Both of these chemical processes may affect the functions of T helper/regulatory cells that are crucial for tempering the inflammatory response after a viral infection.

  11. Prevalence of African swine fever virus and classical swine fever virus antibodies in pigs in Benue State, Nigeria.

    PubMed

    Asambe, A; Sackey, A K B; Tekdek, L B

    2018-03-01

    This study investigated the prevalence of African swine fever virus (ASFV) and classical swine fever virus (CSFV) antibodies in pigs in Benue State, Nigeria. Serum samples were collected from a total of 460 pigs, including 416 from 74 piggeries and 44 from Makurdi slaughter slab. The samples were analysed using indirect enzyme-linked immunosorbent assay (ELISA) test kit to detect the presence of ASFV antibodies, while competitive ELISA test kit was used to detect antibodies to CSFV. Our findings showed a total ASF prevalence of 13 (2.8%), while prevalences of 7 (1.7%) and 6 (13.6%) were observed in piggeries and in Makurdi slaughter slab, respectively. However, no CSFV antibody sera were detected in this study. Relatively higher ASFV antibody-positive pigs were detected in the slaughter slab than in piggeries. The difference in prevalence of ASF between the two locations was significantly associated (p = 0.017). These findings suggest the presence of ASFV antibody-positive pig in Benue State, Nigeria. Continuous surveillance and monitoring of these diseases among pigs in Nigeria to prevent any fulminating outbreak are recommended.

  12. Development of a competitive ELISA using a truncated E2 recombinant protein as antigen for detection of antibodies to classical swine fever virus.

    PubMed

    Clavijo, A; Lin, M; Riva, J; Mallory, M; Lin, F; Zhou, E M

    2001-02-01

    The sequence encoding a truncated E2 glycoprotein of the Alfort/187 strain of classical swine fever virus (CSFV) was expressed in Escherichia coli using the pET expression system and the recombinant product purified by Ni-NTA agarose affinity chromatography. The antigenicity of this recombinant protein was demonstrated by immunoblot using anti- CSFV-specific antibodies. A monoclonal antibody was produced against the truncated E2 protein and used as competitor in an ELISA for the detection of antibodies to CSFV. Specific antibodies were demonstrated by competitive ELISA (C-ELISA) as early as 21 days post-infection (dpi) in experimentally infected pigs. Seroconversion was demonstrated by C-ELISA and neutralising peroxidase-linked assay (NPLA) in all infected animals by 4 weeks. No cross-reaction with antibodies to bovine viral diarrhoea virus (BVDV) was seen in the C-ELISA using sera from experimentally infected pigs. The C-ELISA is not intended as a substitute for the NPLA. However, it is expected it will be useful for monitoring and prevalence studies. It will also assist in testing a large number of samples in the event of an outbreak. Copyright 2001 Harcourt Publishers Ltd.

  13. Safety and immunogenicity of a gE/gI/TK gene-deleted pseudorabies virus variant expressing the E2 protein of classical swine fever virus in pigs.

    PubMed

    Lei, Jian-Lin; Xia, Shui-Li; Wang, Yimin; Du, Mingliang; Xiang, Guang-Tao; Cong, Xin; Luo, Yuzi; Li, Lian-Feng; Zhang, Lingkai; Yu, Jiahui; Hu, Yonghao; Qiu, Hua-Ji; Sun, Yuan

    2016-06-01

    Classical swine fever (CSF) and pseudorabies (PR) are both major infectious diseases of pigs, causing enormous economic losses to the swine industry in many countries. A marker vaccine that enables differentiation of infected from vaccinated animals (DIVA) is highly desirable for control and eradication of these two diseases in endemic areas. Since late 2011, PR outbreaks have been frequently reported in many Bartha-K61-vaccinated pig farms in China. It has been demonstrated that a pseudorabies virus (PRV) variant with altered antigenicity and increased pathogenicity was responsible for the outbreaks. Previously, we showed that rPRVTJ-delgE/gI/TK, a gE/gI/TK-deleted PRV variant, was safe for susceptible animals and provided a complete protection against lethal PRV variant challenge, indicating that rPRVTJ-delgE/gI/TK can be used as an attractive vaccine vector. To develop a safe bivalent vaccine against CSF and PR, we generated a recombinant virus rPRVTJ-delgE/gI/TK-E2 expressing the E2 protein of classical swine fever virus (CSFV) based on rPRVTJ-delgE/gI/TK and evaluated its safety and immunogenicity in pigs. The results indicated that pigs (n=5) immunized with rPRVTJ-delgE/gI/TK-E2 of different doses did not exhibit clinical signs or viral shedding following immunization, the immunized pigs produced anti-PRV or anti-CSFV neutralizing antibodies and the pigs immunized with 10(6) or 10(5) TCID50 rPRVTJ-delgE/gI/TK-E2 were completely protected against the lethal challenge with either CSFV Shimen strain or variant PRV TJ strain. These findings suggest that rPRVTJ-delgE/gI/TK-E2 is a promising bivalent DIVA vaccine candidate against CSFV and PRV coinfections. Copyright © 2016 European Federation of Immunological Societies. Published by Elsevier B.V. All rights reserved.

  14. Characterization of Helper Virus-Independent Cytopathogenic Classical Swine Fever Virus Generated by an In Vivo RNA Recombination System

    PubMed Central

    Gallei, Andreas; Rümenapf, Till; Thiel, Heinz-Jürgen; Becher, Paul

    2005-01-01

    Molecular analyses revealed that most cytopathogenic (cp) pestivirus strains evolve from noncytopathogenic (noncp) viruses by nonhomologous RNA recombination. In contrast to bovine viral diarrhea virus (BVDV), cp classical swine fever virus (CSFV) field isolates were rarely detected and always represented helper virus-dependent subgenomes. To investigate RNA recombination in more detail, we recently established an in vivo system allowing the efficient generation of recombinant cp BVDV strains in cell culture after transfecting a synthetic subgenomic and nonreplicatable transcript into cells being infected with noncp BVDV (A. Gallei, A. Pankraz, H.-J. Thiel, and P. Becher, J. Virol. 78:6271-6281, 2004). Using an analogous approach, the first helper virus-independent cp CSFV strain (CP G1) has now been generated by RNA recombination. Accordingly, this study demonstrates the applicability of RNA recombination for designing new viral RNA genomes. The genomic RNA of CP G1 has a calculated size of 18.139 kb, almost 6 kb larger than all previously described CSFV genomes. It contains cellular sequences encoding a polyubiquitin fragment directly upstream of the nonstructural protein NS3 coding gene together with a duplication of viral sequences. CP G1 induces a cytopathic effect on different tissue culture cell lines from pigs and cattle. Subsequent analyses addressed growth kinetics, expression of NS3, and genetic stability of CP G1. PMID:15681445

  15. Development of a duplex lateral flow assay for simultaneous detection of antibodies against African and Classical swine fever viruses.

    PubMed

    Sastre, Patricia; Pérez, Teresa; Costa, Sofia; Yang, Xiaoping; Räber, Alex; Blome, Sandra; Goller, Katja V; Gallardo, Carmina; Tapia, Istar; García, Julia; Sanz, Antonio; Rueda, Paloma

    2016-09-01

    Classical swine fever (CSF) and African swine fever (ASF) are both highly contagious diseases of domestic pigs and wild boar and are clinically indistinguishable. For both diseases, antibody detection is an integral and crucial part of prevention and control measures. The purpose of our study was to develop and initially validate a duplex pen-side test for simultaneous detection and differentiation of specific antibodies against CSF virus (CSFV) and ASF virus (ASFV). The test was based on the major capsid protein VP72 of ASFV and the structural protein E2 of CSFV, both considered the most immunogenic proteins of these viruses. The performance of the pen-side test was evaluated using a panel of porcine samples consisting of experimental, reference, and field sera, with the latter collected from European farms free of both diseases. The new lateral flow assay was able to detect specific antibodies to ASFV or CSFV, showing good levels of sensitivity and specificity. These preliminary data indicate the potential of the newly developed pen-side test for rapid differential detection of antibodies found in the 2 diseases, which is of particular importance in the field and in front-line laboratories where equipment and skilled personnel are limited and control of ASF and CSF is crucial. © 2016 The Author(s).

  16. A Review of Classical Swine Fever Virus and Routes of Introduction into the United States and the Potential for Virus Establishment.

    PubMed

    Brown, Vienna R; Bevins, Sarah N

    2018-01-01

    Classical swine fever (CSF) is caused by CSF virus (CSFV) which can be the source of substantial morbidity and mortality events in affected swine. The disease can take one of several forms (acute, chronic, or prenatal) and depending on the virulence of the inoculating strain may result in a lethal infection irrespective of the form acquired. Because of the disease-free status of the United States and the high cost of a viral incursion, a summary of US vulnerabilities for viral introduction and persistence is provided. The legal importation of live animals as well as animal products, byproducts, and animal feed serve as a potential route of viral introduction. Current import regulations are described as are mitigation strategies that are commonly utilized to prevent pathogens, including CSFV, from entering the US. The illegal movement of suids and their products as well as an event of bioterrorism are both feasible routes of viral introduction but are difficult to restrict or regulate. Ultimately, recommendations are made for data that would be useful in the event of a viral incursion. Population and density mapping for feral swine across the United States would be valuable in the event of a viral introduction or spillover; density data could further contribute to understanding the risk of infection in domestic swine. Additionally, ecological and behavioral studies, including those that evaluate the effects of anthropogenic food sources that support feral swine densities far above the carrying capacity would provide invaluable insight to our understanding of how human interventions affect feral swine populations. Further analyses to determine the sampling strategies necessary to detect low levels of antibody prevalence in feral swine would also be valuable.

  17. A Review of Classical Swine Fever Virus and Routes of Introduction into the United States and the Potential for Virus Establishment

    PubMed Central

    Brown, Vienna R.; Bevins, Sarah N.

    2018-01-01

    Classical swine fever (CSF) is caused by CSF virus (CSFV) which can be the source of substantial morbidity and mortality events in affected swine. The disease can take one of several forms (acute, chronic, or prenatal) and depending on the virulence of the inoculating strain may result in a lethal infection irrespective of the form acquired. Because of the disease-free status of the United States and the high cost of a viral incursion, a summary of US vulnerabilities for viral introduction and persistence is provided. The legal importation of live animals as well as animal products, byproducts, and animal feed serve as a potential route of viral introduction. Current import regulations are described as are mitigation strategies that are commonly utilized to prevent pathogens, including CSFV, from entering the US. The illegal movement of suids and their products as well as an event of bioterrorism are both feasible routes of viral introduction but are difficult to restrict or regulate. Ultimately, recommendations are made for data that would be useful in the event of a viral incursion. Population and density mapping for feral swine across the United States would be valuable in the event of a viral introduction or spillover; density data could further contribute to understanding the risk of infection in domestic swine. Additionally, ecological and behavioral studies, including those that evaluate the effects of anthropogenic food sources that support feral swine densities far above the carrying capacity would provide invaluable insight to our understanding of how human interventions affect feral swine populations. Further analyses to determine the sampling strategies necessary to detect low levels of antibody prevalence in feral swine would also be valuable. PMID:29556501

  18. A multiplex reverse transcription PCR and automated electronic microarray assay for detection and differentiation of seven viruses affecting swine.

    PubMed

    Erickson, A; Fisher, M; Furukawa-Stoffer, T; Ambagala, A; Hodko, D; Pasick, J; King, D P; Nfon, C; Ortega Polo, R; Lung, O

    2018-04-01

    Microarray technology can be useful for pathogen detection as it allows simultaneous interrogation of the presence or absence of a large number of genetic signatures. However, most microarray assays are labour-intensive and time-consuming to perform. This study describes the development and initial evaluation of a multiplex reverse transcription (RT)-PCR and novel accompanying automated electronic microarray assay for simultaneous detection and differentiation of seven important viruses that affect swine (foot-and-mouth disease virus [FMDV], swine vesicular disease virus [SVDV], vesicular exanthema of swine virus [VESV], African swine fever virus [ASFV], classical swine fever virus [CSFV], porcine respiratory and reproductive syndrome virus [PRRSV] and porcine circovirus type 2 [PCV2]). The novel electronic microarray assay utilizes a single, user-friendly instrument that integrates and automates capture probe printing, hybridization, washing and reporting on a disposable electronic microarray cartridge with 400 features. This assay accurately detected and identified a total of 68 isolates of the seven targeted virus species including 23 samples of FMDV, representing all seven serotypes, and 10 CSFV strains, representing all three genotypes. The assay successfully detected viruses in clinical samples from the field, experimentally infected animals (as early as 1 day post-infection (dpi) for FMDV and SVDV, 4 dpi for ASFV, 5 dpi for CSFV), as well as in biological material that were spiked with target viruses. The limit of detection was 10 copies/μl for ASFV, PCV2 and PRRSV, 100 copies/μl for SVDV, CSFV, VESV and 1,000 copies/μl for FMDV. The electronic microarray component had reduced analytical sensitivity for several of the target viruses when compared with the multiplex RT-PCR. The integration of capture probe printing allows custom onsite array printing as needed, while electrophoretically driven hybridization generates results faster than conventional microarrays that rely on passive hybridization. With further refinement, this novel, rapid, highly automated microarray technology has potential applications in multipathogen surveillance of livestock diseases. © 2017 Her Majesty the Queen in Right of Canada • Transboundary and Emerging Diseases.

  19. Characterization of a new chimeric marker vaccine candidate with a mutated antigenic E2-epitope.

    PubMed

    Reimann, Ilona; Depner, Klaus; Utke, Katrin; Leifer, Immanuel; Lange, Elke; Beer, Martin

    2010-04-21

    A new chimeric pestivirus "CP7_E1E2alf_TLA", based on the infectious cDNA of bovine viral diarrhea virus (BVDV) strain CP7, was constructed. The substitution of BVDV E1 and E2 with the respective proteins of classical swine fever (CSF) strain Alfort 187 allows an optimal heterodimerization of E1 and E2 in the chimeric virus, which is beneficial for efficient and authentic virus assembly and growth. In addition, for implementation of E2-based marker diagnostics, the previously described antigenic CSFV-specific TAVSPTTLR epitope was exchanged with the corresponding E2-epitope of BVDV strain CP7. Recombinant virus CP7_E1E2alf_TLA displayed a growth defect, and was not reacting with monoclonal antibodies used in commercial E2 antibody blocking ELISAs. Therefore, efficacy as well as marker properties of CP7_E1E2alf_TLA were investigated in an animal experiment with both a high dose and a low dose vaccine preparation. All CP7_E1E2alf_TLA-vaccinated animals seroconverted until day 28 post-vaccination with neutralizing antibodies. Furthermore, at the day of challenge infection CP7_E1E2alf_TLA-immunized animals showed distinct lower ELISA values in a commercial CSFV E2 antibody test in comparison to the C-strain vaccinated controls. However, E2-ELISA reactivity as well as neutralizing titers were directly connected to the dosage used for vaccination, and only the low dose group had E2-ELISA values below threshold until challenge infection. Following challenge infection with highly virulent CSFV strain Koslov, all vaccinees were protected, however, short-term fever episodes and very limited CSFV genome detection with very low copy numbers could be observed. In conclusion, manipulation of the TAVSPTTLR-epitope within the tested chimeric virus resulted in an slightly reduced efficacy, but the E2 marker properties unexpectedly did not allow a clear differentiation of infected from vaccinated animals in some cases. Copyright 2009 Elsevier B.V. All rights reserved.

  20. Complex Virus–Host Interactions Involved in the Regulation of Classical Swine Fever Virus Replication: A Minireview

    PubMed Central

    Li, Su; Wang, Jinghan; Yang, Qian; Naveed Anwar, Muhammad; Yu, Shaoxiong; Qiu, Hua-Ji

    2017-01-01

    Classical swine fever (CSF), caused by classical swine fever virus (CSFV), is one of the most devastating epizootic diseases of pigs in many countries. Viruses are small intracellular parasites and thus rely on the cellular factors for replication. Fundamental aspects of CSFV–host interactions have been well described, such as factors contributing to viral attachment, modulation of genomic replication and translation, antagonism of innate immunity, and inhibition of cell apoptosis. However, those host factors that participate in the viral entry, assembly, and release largely remain to be elucidated. In this review, we summarize recent progress in the virus–host interactions involved in the life cycle of CSFV and analyze the potential mechanisms of viral entry, assembly, and release. We conclude with future perspectives and highlight areas that require further understanding. PMID:28678154

  1. A single dose of the novel chimeric subunit vaccine E2-CD154 confers early full protection against classical swine fever virus.

    PubMed

    Suárez, Marisela; Sordo, Yusmel; Prieto, Yanet; Rodríguez, María P; Méndez, Lídice; Rodríguez, Elsa M; Rodríguez-Mallon, Alina; Lorenzo, Elianet; Santana, Elaine; González, Nemecio; Naranjo, Paula; Frías, María Teresa; Carpio, Yamila; Estrada, Mario Pablo

    2017-08-03

    Classical swine fever is an economically important, highly contagious disease of swine worldwide. Subunit vaccines are a suitable alternative for the control of classical swine fever. However, such vaccines have as the main drawback the relatively long period of time required to induce a protective response, which hampers their use under outbreak conditions. In this work, a lentivirus-based gene delivery system is used to obtain a stable recombinant HEK 293 cell line for the expression of E2-CSFV antigen fused to porcine CD154 as immunostimulant molecule. The E2-CD154 chimeric protein was secreted into the medium by HEK293 cells in a concentration around 50mg/L in suspension culture conditions using spinner bottles. The E2-CD154 immunized animals were able to overcome the challenge with a high virulent CSF virus strain performed 7days after a unique dose of the vaccine without clinical manifestations of the disease. Specific anti-CSFV neutralizing antibodies and IFN-γ were induced 8days after challenge equivalent to 14days post-vaccination. The present work constitutes the first report of a subunit vaccine able to confer complete protection by the end of the first week after a single vaccination. These results suggest that the E2-CD154 antigen could be potentially used under outbreak conditions to stop CSFV spread and for eradication programs in CSF enzootic areas. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Effect of specific amino acid substitutions in the putative fusion peptide of structural glycoprotein E2 on Classical Swine Fever Virus replication

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Fernández-Sainz, I.J.; Largo, E.; Gladue, D.P.

    E2, along with E{sup rns} and E1, is an envelope glycoprotein of Classical Swine Fever Virus (CSFV). E2 is involved in several virus functions: cell attachment, host range susceptibility and virulence in natural hosts. Here we evaluate the role of a specific E2 region, {sup 818}CPIGWTGVIEC{sup 828}, containing a putative fusion peptide (FP) sequence. Reverse genetics utilizing a full-length infectious clone of the highly virulent CSFV strain Brescia (BICv) was used to evaluate how individual amino acid substitutions within this region of E2 may affect replication of BICv. A synthetic peptide representing the complete E2 FP amino acid sequence adoptedmore » a β-type extended conformation in membrane mimetics, penetrated into model membranes, and perturbed lipid bilayer integrity in vitro. Similar peptides harboring amino acid substitutions adopted comparable conformations but exhibited different membrane activities. Therefore, a preliminary characterization of the putative FP {sup 818}CPIGWTGVIEC{sup 828} indicates a membrane fusion activity and a critical role in virus replication. - Highlights: • A putative fusion peptide (FP) region in CSFV E2 protein was shown to be critical for virus growth. • Synthetic FPs were shown to efficiently penetrate into lipid membranes using an in vitro model. • Individual residues in the FP affecting virus replication were identified by reverse genetics. • The same FP residues are also responsible for mediating membrane fusion.« less

  3. Detection of African swine fever, classical swine fever, and foot-and-mouth disease viruses in swine oral fluids by multiplex reverse transcription real-time polymerase chain reaction.

    PubMed

    Grau, Frederic R; Schroeder, Megan E; Mulhern, Erin L; McIntosh, Michael T; Bounpheng, Mangkey A

    2015-03-01

    African swine fever (ASF), classical swine fever (CSF), and foot-and-mouth disease (FMD) are highly contagious animal diseases of significant economic importance. Pigs infected with ASF and CSF viruses (ASFV and CSFV) develop clinical signs that may be indistinguishable from other diseases. Likewise, various causes of vesicular disease can mimic clinical signs caused by the FMD virus (FMDV). Early detection is critical to limiting the impact and spread of these disease outbreaks, and the ability to perform herd-level surveillance for all 3 diseases rapidly and cost effectively using a single diagnostic sample and test is highly desirable. This study assessed the feasibility of simultaneous ASFV, CSFV, and FMDV detection by multiplex reverse transcription real-time polymerase chain reaction (mRT-qPCR) in swine oral fluids collected through the use of chewing ropes. Animal groups were experimentally infected independently with each virus, observed for clinical signs, and oral fluids collected and tested throughout the course of infection. All animal groups chewed on the ropes readily before and after onset of clinical signs and before onset of lameness or serious clinical signs. ASFV was detected as early as 3 days postinoculation (dpi), 2-3 days before onset of clinical disease; CSFV was detected at 5 dpi, coincident with onset of clinical disease; and FMDV was detected as early as 1 dpi, 1 day before the onset of clinical disease. Equivalent results were observed in 4 independent studies and demonstrate the feasibility of oral fluids and mRT-qPCR for surveillance of ASF, CSF, and FMD in swine populations. © 2015 The Author(s).

  4. Factors affecting the infectivity of tissues from pigs with classical swine fever: thermal inactivation rates and oral infectious dose.

    PubMed

    Cowan, Lucie; Haines, Felicity J; Everett, Helen E; Crudgington, Bentley; Johns, Helen L; Clifford, Derek; Drew, Trevor W; Crooke, Helen R

    2015-03-23

    Outbreaks of classical swine fever are often associated with ingestion of pig meat or products derived from infected pigs. Assessment of the disease risks associated with material of porcine origin requires knowledge on the likely amount of virus in the original material, how long the virus may remain viable within the resulting product and how much of that product would need to be ingested to result in infection. Using material from pigs infected with CSFV, we determined the viable virus concentrations in tissues that comprise the majority of pork products. Decimal reduction values (D values), the time required to reduce the viable virus load by 90% (or 1 log10), were determined at temperatures of relevance for chilling, cooking, composting and ambient storage. The rate of CSFV inactivation varied in different tissues. At lower temperatures, virus remained viable for substantially longer in muscle and serum compared to lymphoid and fat tissues. To enable estimation of the temperature dependence of inactivation, the temperature change required to change the D values by 90% (Z values) were determined as 13 °C, 14 °C, 12 °C and 10 °C for lymph node, fat, muscle and serum, respectively. The amount of virus required to infect 50% of pigs by ingestion was determined by feeding groups of animals with moderately and highly virulent CSFV. Interestingly, the virulent virus did not initiate infection at a lower dose than the moderately virulent strain. Although higher than for intranasal inoculation, the amount of virus required for infection via ingestion is present in only a few grams of tissue from infected animals. Crown Copyright © 2015. Published by Elsevier B.V. All rights reserved.

  5. Sorting out pestiviral phylogeny: A tale of viral swarms, red herrings, and sons of Bs

    USDA-ARS?s Scientific Manuscript database

    Initially three species, border disease virus (BDV), bovine viral diarrhea virus (BVDV), and classical swine fever virus (CSFV), were recognized in the pestivirus genus. These three species were defined by their host of origin, and to a lesser extent by clinical presentation. Subsequently, attempts ...

  6. HoBi-like viruses: an emerging group of pestiviruses

    USDA-ARS?s Scientific Manuscript database

    The genus Pestivirus is composed by four important pathogens of livestock: bovine viral diarrhea virus types 1 and 2 (BVDV-1 and BVDV-2), classical swine fever virus (CSFV) and border disease virus of sheep (BDV). BVDV are major pathogens of cattle and infection results in significant economic losse...

  7. Pore-forming activity of pestivirus p7 in a minimal model system supports genus-specific viroporin function

    USDA-ARS?s Scientific Manuscript database

    Viroporins are small integral membrane proteins functional in viral assembly and egress by promoting permeabilization. Blocking of viroporin function therefore constitutes a target for antiviral development. Classical swine fever virus (CSFV) protein p7 has been recently regarded as a class II viro...

  8. Clinical presentation resembling mucosal disease associated with 'HoBi'-like pestivirus in a field outbreak

    USDA-ARS?s Scientific Manuscript database

    The genus Pestivirus of the family Flaviviridae consists of four recognized species: Bovine viral diarrhea virus 1 (BVDV-1), Bovine viral diarrhea virus 2 (BVDV-2), Classical swine fever virus (CSFV) And Border disease virus (BDV). Recently, atypical pestiviruses (‘HoBi’-like pestiviruses) were iden...

  9. The E2 glycoprotein of classical swine fever virus is a virulence determinant in swine.

    PubMed

    Risatti, G R; Borca, M V; Kutish, G F; Lu, Z; Holinka, L G; French, R A; Tulman, E R; Rock, D L

    2005-03-01

    To identify genetic determinants of classical swine fever virus (CSFV) virulence and host range, chimeras of the highly pathogenic Brescia strain and the attenuated vaccine strain CS were constructed and evaluated for viral virulence in swine. Upon initial screening, only chimeras 138.8v and 337.14v, the only chimeras containing the E2 glycoprotein of CS, were attenuated in swine despite exhibiting unaltered growth characteristics in primary porcine macrophage cell cultures. Additional viral chimeras were constructed to confirm the role of E2 in virulence. Chimeric virus 319.1v, which contained only the CS E2 glycoprotein in the Brescia background, was markedly attenuated in pigs, exhibiting significantly decreased virus replication in tonsils, a transient viremia, limited generalization of infection, and decreased virus shedding. Chimeras encoding all Brescia structural proteins in a CS genetic background remained attenuated, indicating that additional mutations outside the structural region are important for CS vaccine virus attenuation. These results demonstrate that CS E2 alone is sufficient for attenuating Brescia, indicating a significant role for the CSFV E2 glycoprotein in swine virulence.

  10. The influence of viral coding sequences on pestivirus IRES activity reveals further parallels with translation initiation in prokaryotes.

    PubMed Central

    Fletcher, Simon P; Ali, Iraj K; Kaminski, Ann; Digard, Paul; Jackson, Richard J

    2002-01-01

    Classical swine fever virus (CSFV) is a member of the pestivirus family, which shares many features in common with hepatitis C virus (HCV). It is shown here that CSFV has an exceptionally efficient cis-acting internal ribosome entry segment (IRES), which, like that of HCV, is strongly influenced by the sequences immediately downstream of the initiation codon, and is optimal with viral coding sequences in this position. Constructs that retained 17 or more codons of viral coding sequence exhibited full IRES activity, but with only 12 codons, activity was approximately 66% of maximum in vitro (though close to maximum in transfected BHK cells), whereas with just 3 codons or fewer, the activity was only approximately 15% of maximum. The minimal coding region elements required for high activity were exchanged between HCV and CSFV. Although maximum activity was observed in each case with the homologous combination of coding region and 5' UTR, the heterologous combinations were sufficiently active to rule out a highly specific functional interplay between the 5' UTR and coding sequences. On the other hand, inversion of the coding sequences resulted in low IRES activity, particularly with the HCV coding sequences. RNA structure probing showed that the efficiency of internal initiation of these chimeric constructs correlated most closely with the degree of single-strandedness of the region around and immediately downstream of the initiation codon. The low activity IRESs could not be rescued by addition of supplementary eIF4A (the initiation factor with ATP-dependent RNA helicase activity). The extreme sensitivity to secondary structure around the initiation codon is likely to be due to the fact that the eIF4F complex (which has eIF4A as one of its subunits) is not required for and does not participate in initiation on these IRESs. PMID:12515388

  11. Survival of viral pathogens in animal feed ingredients under transboundary shipping models

    PubMed Central

    Bauermann, Fernando V.; Niederwerder, Megan C.; Singrey, Aaron; Clement, Travis; de Lima, Marcelo; Long, Craig; Patterson, Gilbert; Sheahan, Maureen A.; Stoian, Ana M. M.; Petrovan, Vlad; Jones, Cassandra K.; De Jong, Jon; Ji, Ju; Spronk, Gordon D.; Minion, Luke; Christopher-Hennings, Jane; Zimmerman, Jeff J.; Rowland, Raymond R. R.; Nelson, Eric; Sundberg, Paul; Diel, Diego G.

    2018-01-01

    The goal of this study was to evaluate survival of important viral pathogens of livestock in animal feed ingredients imported daily into the United States under simulated transboundary conditions. Eleven viruses were selected based on global significance and impact to the livestock industry, including Foot and Mouth Disease Virus (FMDV), Classical Swine Fever Virus (CSFV), African Swine Fever Virus (ASFV), Influenza A Virus of Swine (IAV-S), Pseudorabies virus (PRV), Nipah Virus (NiV), Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), Swine Vesicular Disease Virus (SVDV), Vesicular Stomatitis Virus (VSV), Porcine Circovirus Type 2 (PCV2) and Vesicular Exanthema of Swine Virus (VESV). Surrogate viruses with similar genetic and physical properties were used for 6 viruses. Surrogates belonged to the same virus families as target pathogens, and included Senecavirus A (SVA) for FMDV, Bovine Viral Diarrhea Virus (BVDV) for CSFV, Bovine Herpesvirus Type 1 (BHV-1) for PRV, Canine Distemper Virus (CDV) for NiV, Porcine Sapelovirus (PSV) for SVDV and Feline Calicivirus (FCV) for VESV. For the remaining target viruses, actual pathogens were used. Virus survival was evaluated using Trans-Pacific or Trans-Atlantic transboundary models involving representative feed ingredients, transport times and environmental conditions, with samples tested by PCR, VI and/or swine bioassay. SVA (representing FMDV), FCV (representing VESV), BHV-1 (representing PRV), PRRSV, PSV (representing SVDV), ASFV and PCV2 maintained infectivity during transport, while BVDV (representing CSFV), VSV, CDV (representing NiV) and IAV-S did not. Notably, more viruses survived in conventional soybean meal, lysine hydrochloride, choline chloride, vitamin D and pork sausage casings. These results support published data on transboundary risk of PEDV in feed, demonstrate survival of certain viruses in specific feed ingredients (“high-risk combinations”) under conditions simulating transport between continents and provide further evidence that contaminated feed ingredients may represent a risk for transport of pathogens at domestic and global levels. PMID:29558524

  12. Alteration of a Second Putative Fusion Peptide of Structural Glycoprotein E2 of Classical Swine Fever Virus Alters Virus Replication and Virulence in Swine

    PubMed Central

    Holinka, L. G.; Largo, E.; Gladue, D. P.; O'Donnell, V.; Risatti, G. R.; Nieva, J. L.

    2016-01-01

    ABSTRACT E2, the major envelope glycoprotein of classical swine fever virus (CSFV), is involved in several critical virus functions, including cell attachment, host range susceptibility, and virulence in natural hosts. Functional structural analysis of E2 based on a Wimley-White interfacial hydrophobicity distribution predicted the involvement of a loop (residues 864 to 881) stabilized by a disulfide bond (869CKWGGNWTCV878, named FPII) in establishing interactions with the host cell membrane. This loop further contains an 872GG873 dipeptide, as well as two aromatic residues (871W and 875W) accessible to solvent. Reverse genetics utilizing a full-length infectious clone of the highly virulent CSFV strain Brescia (BICv) was used to evaluate how amino acid substitutions within FPII may affect replication of BICv in vitro and virus virulence in swine. Recombinant CSFVs containing mutations in different residues of FPII were constructed. A particular construct, harboring amino acid substitutions W871T, W875D, and V878T (FPII.2), demonstrated a significantly decreased ability to replicate in a swine cell line (SK6) and swine macrophage primary cell cultures. Interestingly, mutated virus FPII.2 was completely attenuated in pigs. Also, animals infected with FPII.2 virus were protected against virulent challenge with Brescia virus at 21 days postvaccination. Supporting a role for the E2 the loop from residues 864 to 881 in membrane fusion, only synthetic peptides that were based on the native E2 functional sequence were competent for insertion into model membranes and perturbation of their integrity, and this functionality was lost in synthetic peptides harboring amino acid substitutions W871T, W875D, and V878T in FPII.2. IMPORTANCE This report describes the identification and characterization of a putative fusion peptide (FP) in the major structural protein E2 of classical swine fever virus (CSFV). The FP identification was performed by functional structural analysis of E2. We characterized the functional significance of this FP by using artificial membranes. Replacement of critical amino acid residues within the FP radically alters how it interacts with the artificial membranes. When we introduced the same mutations into the viral sequence, there was a reduction in replication in cell cultures, and when we infected domestic swine, the natural host of CSFV host, we observed that the virus was now completely attenuated in swine. In addition, the virus mutant that was attenuated in vivo efficiently protected pigs against wild-type virus. These results provide the proof of principle to support as a strategy for vaccine development the discovery and manipulation of FPs. PMID:27605674

  13. Interaction of structural core protein of Classical Swine Fever Virus with endoplasmic reticulum-associated degradation pathway protein OS9

    USDA-ARS?s Scientific Manuscript database

    Classical Swine Fever Virus (CSFV) Core protein is involved in virus RNA protection, transcription regulation and virus virulence. To discover additional Core protein functions a yeast two-hybrid system was used to identify host proteins that interact with Core. Among the identified host proteins, t...

  14. Alteration of a second putative fusion peptide of structural glycoprotein E2 of Classical Swine Fever Virus alters virus replication and virulence in swine

    USDA-ARS?s Scientific Manuscript database

    E2, the major envelope glycoprotein of Classical Swine Fever Virus (CSFV), is involved in several critical virus functions including cell attachment, host range susceptibility, and virulence in natural hosts. Functional structural analysis of E2 based on Wimley-White interfacial hydrophobicity dis...

  15. Classical Swine Fever Virus p7 protein is a viroporin involved in virulence in swine

    USDA-ARS?s Scientific Manuscript database

    The non-structural protein p7 of Classical Swine Fever Virus (CSFV) is a hydrophobic polypeptide with an apparent molecular mass of 7 kDa. The protein contains two hydrophobic stretches of amino acids interrupted by a short charged segment that are predicted to form transmembrane helices and a cytos...

  16. Simulating the spread of classical swine fever virus between a hypothetical wild-boar population and domestic pig herds in Denmark.

    PubMed

    Boklund, A; Goldbach, S G; Uttenthal, A; Alban, L

    2008-07-15

    Denmark has no free-range wild-boar population. However, Danish wildlife organizations have suggested that wild boar should be reintroduced into the wild to broaden national biodiversity. Danish pig farmers fear that this would lead to a higher risk of introduction of classical swine fever virus (CSFV), which could have enormous consequences in terms of loss of pork exports. We conducted a risk assessment to address the additional risk of introducing and spreading CSFV due to the reintroduction of wild boar. In this paper, we present the part of the risk assessment that deals with the spread of CSFV between the hypothetical wild-boar population and the domestic population. Furthermore, the economic impact is assessed taking the perspective of the Danish national budget and the Danish pig industry. We used InterSpreadPlus to model the differential classical swine fever (CSF) risk due to wild boar. Nine scenarios were run to elucidate the effect of: (a) presence of wild boar (yes/no), (b) locations for the index case (domestic pig herd/wild-boar group), (c) type of control strategy for wild boar (hunting/vaccination) and (d) presence of free-range domestic pigs. The presence of free-range wild boar was simulated in two large forests using data from wildlife studies and Danish habitat data. For each scenario, we estimated (1) the control costs borne by the veterinary authorities, (2) the control-related costs to farmers and (3) the loss of exports associated with an epidemic. Our simulations predict that CSFV will be transmitted from the domestic pig population to wild boar if the infected domestic pig herd is located close to an area with wild boar (<5 km). If an outbreak begins in the wild-boar population, the epidemic will last longer and will occasionally lead to several epidemics because of periodic transfer of virus from groups of infected wild boar to domestic pig herds. The size and duration of the epidemic will be reduced if there are no free-range domestic pig herds in the area with CSF-infected wild boar. The economic calculations showed that the total national costs for Denmark (i.e. the direct costs to the national budget and the costs to the pig industry) related to an outbreak of CSF in Denmark will be highly driven by the reactions of the export markets and in particular of the non-EU markets. Unfortunately, there is a substantial amount of uncertainty surrounding this issue. If hunting is used as a control measure, the average expenses related to a CSF outbreak will be 40% higher if wild boar are present compared with not present. However, a vaccination strategy for wild boar will double the total costs compared with a hunting strategy.

  17. Effect of specific amino acid substitutions in the putative fusion peptide of structural glycoprotein E2 on Classical Swine Fever Virus replication

    USDA-ARS?s Scientific Manuscript database

    E2, along with E^rns and E1, is an envelope glycoprotein of Classical Swine Fever Virus (CSFV). E2 is involved in several virus functions including cell attachment, host range susceptibility and virulence in natural hosts. In infected cells, E2 forms homodimers as well as heterodimers with E1, media...

  18. Deploying Crowd-Sourced Formal Verification Systems in a DoD Network

    DTIC Science & Technology

    2013-09-01

    INTENTIONALLY LEFT BLANK 1 I. INTRODUCTION A. INTRODUCTION In 2014 cyber attacks on critical infrastructure are expected to increase...CSFV systems on the Internet‒‒possibly using cloud infrastructure (Dean, 2013). By using Amazon Compute Cloud (EC2) systems, DARPA will use ordinary...through standard access methods. Those clients could be mobile phones, laptops, netbooks, tablet computers or personal digital assistants (PDAs) (Smoot

  19. Mutations in the carboxyl terminal region of E2 glycoprotein of classical swine fever virus are responsible for viral attenuation in swine.

    PubMed

    Risatti, G R; Holinka, L G; Fernandez Sainz, I; Carrillo, C; Kutish, G F; Lu, Z; Zhu, J; Rock, D L; Borca, M V

    2007-08-01

    We have previously reported [Risatti, G.R., Borca, M.V., Kutish, G.F., Lu, Z., Holinka, L.G., French, R.A., Tulman, E.R., Rock, D.L. 2005a. The E2 glycoprotein of classical swine fever virus is a virulence determinant in swine. J. Virol. 79, 3787-3796] that chimeric virus 319.1v containing the E2 glycoprotein gene from Classical Swine Fever Virus (CSFV) vaccine strain CS with the genetic background of highly virulent CSFV strain Brescia (BICv) was markedly attenuated in pigs. To identify the amino acids mediating 319.1v attenuation a series of chimeric viruses containing CS E2 residues in the context of the Brescia strain were constructed. Chimera 357v, containing CS E2 residues 691 to 881 of CSFV polyprotein was virulent, while chimera 358v, containing CS E2 residues 882 to 1064, differing in thirteen amino acids from BICv, was attenuated in swine. Single or double substitutions of those amino acids in BICv E2 to CS E2 residues did not affect virulence. Groups of amino acids were then substituted in BICv E2 to CS E2 residues. Mutant 32v, with six substitutions between residues 975 and 1059, and mutant 33v, with six substitutions between 955 and 994, induced disease indistinguishable from BICv. Mutant 31v, with seven substitutions between residues 882 and 958, induced a delayed onset of lethal disease. Amino acids abrogating BICv virulence were then determined by progressively introducing six CS residues into 31v. Mutant 39v, containing nine residue substitutions, was virulent. Mutant 40v, containing ten residue substitutions, induced mild disease. Mutant 42v, containing twelve substitutions, and mutant 43v, with an amino acid composition identical to 358v, were attenuated in swine indicating that all substitutions were necessary for attenuation of the highly virulent strain Brescia. Importantly, 358v protected swine from challenge with virulent BICv at 3 and 28 days post-infection.

  20. Hepatitis-C-virus-like internal ribosome entry sites displace eIF3 to gain access to the 40S subunit

    NASA Astrophysics Data System (ADS)

    Hashem, Yaser; Des Georges, Amedee; Dhote, Vidya; Langlois, Robert; Liao, Hstau Y.; Grassucci, Robert A.; Pestova, Tatyana V.; Hellen, Christopher U. T.; Frank, Joachim

    2013-11-01

    Hepatitis C virus (HCV) and classical swine fever virus (CSFV) messenger RNAs contain related (HCV-like) internal ribosome entry sites (IRESs) that promote 5'-end independent initiation of translation, requiring only a subset of the eukaryotic initiation factors (eIFs) needed for canonical initiation on cellular mRNAs. Initiation on HCV-like IRESs relies on their specific interaction with the 40S subunit, which places the initiation codon into the P site, where it directly base-pairs with eIF2-bound initiator methionyl transfer RNA to form a 48S initiation complex. However, all HCV-like IRESs also specifically interact with eIF3 (refs 2, 5, 6, 7, 9, 10, 11, 12), but the role of this interaction in IRES-mediated initiation has remained unknown. During canonical initiation, eIF3 binds to the 40S subunit as a component of the 43S pre-initiation complex, and comparison of the ribosomal positions of eIF3 and the HCV IRES revealed that they overlap, so that their rearrangement would be required for formation of ribosomal complexes containing both components. Here we present a cryo-electron microscopy reconstruction of a 40S ribosomal complex containing eIF3 and the CSFV IRES. Remarkably, although the position and interactions of the CSFV IRES with the 40S subunit in this complex are similar to those of the HCV IRES in the 40S-IRES binary complex, eIF3 is completely displaced from its ribosomal position in the 43S complex, and instead interacts through its ribosome-binding surface exclusively with the apical region of domain III of the IRES. Our results suggest a role for the specific interaction of HCV-like IRESs with eIF3 in preventing ribosomal association of eIF3, which could serve two purposes: relieving the competition between the IRES and eIF3 for a common binding site on the 40S subunit, and reducing formation of 43S complexes, thereby favouring translation of viral mRNAs.

  1. HCV-like IRESs displace eIF3 to gain access to the 40S subunit

    PubMed Central

    Hashem, Yaser; des Georges, Amedee; Dhote, Vidya; Langlois, Robert; Liao, Hstau Y.; Grassucci, Robert A.; Pestova, Tatyana V.; Hellen, Christopher U.T.; Frank, Joachim

    2014-01-01

    Hepatitis C virus (HCV) and Classical swine fever virus (CSFV) mRNAs contain related (HCV-like) internal ribosome entry sites (IRESs) that promote 5’-end independent initiation of translation, requiring only a subset of the eukaryotic initiation factors (eIFs) needed for canonical initiation on cellular mRNAs1. Initiation on HCV-like IRESs relies on their specific interaction with the 40S subunit2–8, which places the initiation codon into the P site, where it directly base-pairs with eIF2-bound Met-tRNAiMet to form a 48S initiation complex. However, all HCV-like IRESs also specifically interact with eIF32,5–7,9–12, but the role of this interaction in IRES-mediated initiation has remained unknown. During canonical initiation, eIF3 binds to the 40S subunit as a component of the 43S pre-initiation complex, and comparison of the ribosomal positions of eIF313 and the HCV IRES8 revealed that they overlap, so that their rearrangement would be required for formation of ribosomal complexes containing both components13. Here, we present a cryo-electron microscopy reconstruction of a 40S ribosomal complex containing eIF3 and the CSFV IRES. Strikingly, although the position and interactions of the CSFV IRES with the 40S subunit in this complex are similar to those of the HCV IRES in the 40S/IRES binary complex8, eIF3 is completely displaced from its ribosomal position in the 43S complex, and instead interacts through its ribosome-binding surface exclusively with the apical region of domain III of the IRES. Our results suggest a role for the specific interaction of HCV-like IRESs with eIF3 in preventing ribosomal association of eIF3, which could serve two purposes: relieving the competition between the IRES and eIF3 for a common binding site on the 40S subunit, and reducing formation of 43S complexes, thereby favoring translation of viral mRNAs. PMID:24185006

  2. The E2 glycoprotein is necessary but not sufficient for the adaptation of classical swine fever virus lapinized vaccine C-strain to the rabbit.

    PubMed

    Li, Yongfeng; Xie, Libao; Zhang, Lingkai; Wang, Xiao; Li, Chao; Han, Yuying; Hu, Shouping; Sun, Yuan; Li, Su; Luo, Yuzi; Liu, Lihong; Munir, Muhammad; Qiu, Hua-Ji

    2018-06-01

    Classical swine fever virus (CSFV) C-strain was developed through hundreds of passages of a highly virulent CSFV in rabbits. To investigate the molecular basis for the adaptation of C-strain to the rabbit (ACR), a panel of chimeric viruses with the exchange of glycoproteins E rns , E1, and/or E2 between C-strain and the highly virulent Shimen strain and a number of mutant viruses with different amino acid substitutions in E2 protein were generated and evaluated in rabbits. Our results demonstrate that Shimen-based chimeras expressing E rns -E1-E2, E rns -E2 or E1-E2 but not E rns -E1, E rns , E1, or E2 of C-strain can replicate in rabbits, indicating that E2 in combination with either E rns or E1 confers the ACR. Notably, E2 and the amino acids P108 and T109 in Domain I of E2 are critical in ACR. Collectively, our data indicate that E2 is crucial in mediating the ACR, which requires synergistic contribution of E rns or E1. Copyright © 2018 Elsevier Inc. All rights reserved.

  3. Disease-emergence dynamics and control in a socially-structured wildlife species

    NASA Astrophysics Data System (ADS)

    Pepin, Kim M.; Vercauteren, Kurt C.

    2016-04-01

    Once a pathogen is introduced in a population, key factors governing rate of spread include contact structure, supply of susceptible individuals and pathogen life-history. We examined the interplay of these factors on emergence dynamics and efficacy of disease prevention and response. We contrasted transmission dynamics of livestock viruses with different life-histories in hypothetical populations of feral swine with different contact structures (homogenous, metapopulation, spatial and network). Persistence probability was near 0 for the FMDV-like case under a wide range of parameter values and contact structures, while persistence was probable for the CSFV-like case. There were no sets of conditions where the FMDV-like pathogen persisted in every stochastic simulation. Even when population growth rates were up to 300% annually, the FMDV-like pathogen persisted in <25% of simulations regardless of transmission probabilities and contact structure. For networks and spatial contact structure, persistence probability of the FMDV-like pathogen was always <10%. Because of its low persistence probability, even very early response to the FMDV-like pathogen in feral swine was unwarranted while response to the CSFV-like pathogen was generally effective. When pre-emergence culling of feral swine caused population declines, it was effective at decreasing outbreak size of both diseases by ≥80%.

  4. Substitution of specific cysteine residues in E1 glycoprotein of classical swine fever virus strain Brescia affects formation of E1-E2 heterodimers and alters virulence in swine

    USDA-ARS?s Scientific Manuscript database

    E1, along with E^rns and E2, is one of the three envelope glycoproteins of Classical Swine Fever Virus (CSFV). E1 and E2 are anchored to the virus envelope at their carboxyl termini and E^rns loosely associates with the viral envelope. In infected cells, E2 forms homodimers and heterodimers with E1,...

  5. Validation of two commercial real-time RT-PCR kits for rapid and specific diagnosis of classical swine fever virus.

    PubMed

    Le Dimna, M; Vrancken, R; Koenen, F; Bougeard, S; Mesplède, A; Hutet, E; Kuntz-Simon, G; Le Potier, M F

    2008-01-01

    Two real-time RT-PCR kits, developed by LSI (TaqVet CSF) and ADIAGENE (Adiavet CSF), obtained an agreement to be commercialised in France, subject to conditions, defined by the French Classical Swine Fever (CSF) National Reference Laboratory. The producers were asked to introduce an internal control to check the RNA extraction efficacy. The different criteria assessed were sensitivity, "pestivirus specificity", reproducibility and ease of handling, using 189 different samples. These samples were either CSFV inactivated strains or blood/serum/organs collected from CSFV experimentally infected pigs or naturally infected wild boars. The reproducibility of the assays was confirmed by the analysis of a batch-to-batch panel control that was used for inter-laboratory tests involving nine laboratories. The two kits were also tested for the use in mass diagnostics and the results proved the kits to be suited using pools of blood, serum and tonsils. Moreover, a field evaluation, carried out on spleen samples collected from the CSF surveillance of wild boars in an area known to be infected and from domestic pigs at a slaughterhouse, confirmed the high sensitivity and specificity of the two kits. This step-by-step evaluation procedure confirmed that the two commercial CSF real-time RT-PCR kits have a higher predictive value than the current diagnostic standard, Virus Isolation.

  6. A serosurvey for Brucella suis, classical swine fever virus, porcine circovirus type 2, and pseudorabies virus in feral swine (Sus scrofa) of eastern North Carolina.

    PubMed

    Sandfoss, Mark R; DePerno, Christopher S; Betsill, Carl W; Palamar, Maria Baron; Erickson, Gene; Kennedy-Stoskopf, Suzanne

    2012-04-01

    As feral swine (Sus scrofa) populations expand their range and the opportunity for feral swine hunting increases, there is increased potential for disease transmission that may impact humans, domestic swine, and wildlife. From September 2007 to March 2010, in 13 North Carolina, USA, counties and at Howell Woods Environmental Learning Center, we conducted a serosurvey of feral swine for Brucella suis, pseudorabies virus (PRV), and classical swine fever virus (CSFV); the samples obtained at Howell Woods also were tested for porcine circovirus type 2 (PCV-2). Feral swine serum was collected from trapped and hunter-harvested swine. For the first time since 2004 when screening began, we detected B. suis antibodies in 9% (9/98) of feral swine at Howell Woods and <1% (1/415) in the North Carolina counties. Also, at Howell Woods, we detected PCV-2 antibodies in 59% (53/90) of feral swine. We did not detect antibodies to PRV (n=512) or CSFV (n=307) at Howell Woods or the 13 North Carolina counties, respectively. The detection of feral swine with antibodies to B. suis for the first time in North Carolina warrants increased surveillance of the feral swine population to evaluate speed of disease spread and to establish the potential risk to commercial swine and humans.

  7. Pre-Clinical Evaluation of a Real-Time PCR Assay on a Portable Instrument as a Possible Field Diagnostic Tool: Experiences from the Testing of Clinical Samples for African and Classical Swine Fever Viruses.

    PubMed

    Liu, L; Luo, Y; Accensi, F; Ganges, L; Rodríguez, F; Shan, H; Ståhl, K; Qiu, H-J; Belák, S

    2017-10-01

    African swine fever (ASF) and classical swine fever (CSF) are two highly infectious transboundary animal diseases (TADs) that are serious threats to the pig industry worldwide, including in China, the world's largest pork producer. In this study, a duplex real-time PCR assay was developed for the rapid detection and differentiation of African swine fever virus (ASFV) and classical swine fever virus (CSFV). The assay was performed on a portable, battery-powered PCR thermocycler with a low sample throughput (termed as 'T-COR4 assay'). The feasibility and reliability of the T-COR4 assay as a possible field method was investigated by testing clinical samples collected in China. When evaluated with reference materials or samples from experimental infections, the assay performed in a reliable manner, producing results comparable to those obtained from stationary PCR platforms. Of 59 clinical samples, 41 had results identical to a two-step CSFV real-time PCR assay. No ASFV was detected in these samples. The T-COR4 assay was technically easy to perform and produced results within 3 h, including sample preparation. In combination with a simple sample preparation method, the T-COR4 assay provides a new tool for the field diagnosis and differentiation of ASF and CSF, which could be of particular value in remote areas. © 2016 Blackwell Verlag GmbH.

  8. Comparative analyses of host responses upon infection with moderately virulent classical swine fever virus in domestic pigs and wild boar.

    PubMed

    Petrov, Anja; Blohm, Ulrike; Beer, Martin; Pietschmann, Jana; Blome, Sandra

    2014-07-29

    Classical swine fever (CSF) is one of the most important viral diseases of pigs. Clinical signs may vary from almost inapparent infection to a hemorrhagic fever like illness. Among the host factors leading to different disease courses are age, breed, and immune status. The aim of this study was to compare host responses of different pig breeds upon infection with a recent moderately virulent CSF virus (CSFV) strain, and to assess their impact on the clinical outcome and the efficiency of immune responses. To this means, two domestic pig types (German Landrace and hybrids), were compared to European wild boar. Along with clinical and pathological assessments and routine virological and serological methods, kinetics of immune-cellular parameters were evaluated. All animals were susceptible to infection and despite clinical differences, virus could be detected in all infected animals to similar amounts. All but one animal developed an acute disease course, two landrace animals recovered after a transient infection. One wild boar got chronically infected. Changes in the percentages of lymphocyte subsets in peripheral blood did not show a clear correlation with the clinical outcome. High and early titers of neutralizing antibodies were especially detected in wild boar and German Landrace pigs. While differences among breeds did not have the expected impact on course and outcome of CSFV infection, preload with facultative pathogens and even small differences in age seemed to be more relevant. Future studies will target the characterization of responses observed during different disease courses including cytokine reactions and further analyses of lymphocyte subsets.

  9. Endurance and failure characteristics of modified Vasco X-2, CBS 600 and AISI 9310 spur gears. [aircraft construction materials

    NASA Technical Reports Server (NTRS)

    Townsend, D. P.; Zaretsky, E. V.

    1980-01-01

    Gear endurance tests and rolling-element fatigue tests were conducted to compare the performance of spur gears made from AISI 9310, CBS 600 and modified Vasco X-2 and to compare the pitting fatigue lives of these three materials. Gears manufactured from CBS 600 exhibited lives longer than those manufactured from AISI 9310. However, rolling-element fatigue tests resulted in statistically equivalent lives. Modified Vasco X-2 exhibited statistically equivalent lives to AISI 9310. CBS 600 and modified Vasco X-2 gears exhibited the potential of tooth fracture occurring at a tooth surface fatigue pit. Case carburization of all gear surfaces for the modified Vasco X-2 gears results in fracture at the tips of the gears.

  10. Is there a role for modified probiotics as beneficial microbes: a systematic review of the literature.

    PubMed

    Zorzela, L; Ardestani, S K; McFarland, L V; Vohra, S

    2017-10-13

    Our objective was to conduct a systematic review and meta-analysis for the use of modified (heat-killed or sonicated) probiotics for the efficacy and safety to prevent and treat various diseases. Recent clinical research has focused on living strains of probiotics, but use in high-risk patients and potential adverse reactions including bacteremia has focused interest on alternatives to the use of live probiotics. We searched MEDLINE/PubMed, Embase, Cochrane Central Register of Controlled Trials, CINAHL, Alt Health Watch, Web of Science, Scopus, PubMed, from inception to February 14, 2017 for randomised controlled trials involving modified probiotic strains. The primary outcome was efficacy to prevent or treat disease and the secondary outcome was incidence of adverse events. A total of 40 trials were included (n=3,913): 14 trials (15 arms with modified probiotics and 20 control arms) for the prevention of diseases and 26 trials (29 arms with modified probiotics and 32 control arms) for treatment of various diseases. Modified microbes were compared to either placebo (44%), or the same living probiotic strain (39%) or to only standard therapies (17%). Modified microbes were not significantly more or less effective than the living probiotic in 86% of the preventive trials and 69% of the treatment trials. Modified probiotic strains were significantly more effective in 15% of the treatment trials. Incidence rates of adverse events were similar for modified and living probiotics and other control groups, but many trials did not collect adequate safety data. Although several types of modified probiotics showed significant efficacy over living strains of probiotics, firm conclusions could not be reached due to the limited number of trials using the same type of modified microbe (strain, daily dose and duration) for a specific disease indication. Further research may illuminate other strains of modified probiotics that may have potential as clinical biotherapeutics.

  11. Serologic Survey for Selected Viral and Bacterial Swine Pathogens in Colombian Collared Peccaries ( Pecari tajacu) and Feral Pigs ( Sus scrofa).

    PubMed

    Montenegro, Olga L; Roncancio, Nestor; Soler-Tovar, Diego; Cortés-Duque, Jimena; Contreras-Herrera, Jorge; Sabogal, Sandra; Acevedo, Luz Dary; Navas-Suárez, Pedro Enrique

    2018-06-14

    In South America, wild populations of peccaries coexist with domestic and feral pigs, with poorly understood consequences. We captured 58 collared peccaries ( Pecari tajacu) and 15 feral pigs ( Sus scrofa) in locations of Colombia where coexistence of these species is known. Blood samples were tested for antibodies against four viral agents, classical swine fever virus (CSFV), Aujeszky's disease virus (ADV), porcine circovirus (PCV-2), and vesicular stomatitis virus (New Jersey and Indiana subtypes) and two bacterial agents, Brucella spp. and six serovars of Leptospira interrogans. The prevalence of CSFV was 5% (3/58) in collared peccaries and 7% (1/15) in feral pigs. The prevalence of PCV-2 was 7% (1/15) in collared peccaries and 67% (2/3) in feral pigs. Vesicular stomatitis prevalence was 33% (8/24) in collared peccaries and 67% (4/6) in feral pigs. Leptospira prevalence was 78% (39/50) in collared peccary and 100% (8/8) in feral pigs; bratislava, grippotyphosa, icterohaemorrhagiae, and pomona were the most frequent serovars. Also, the only white-lipped peccary ( Tayassu pecari) sampled was positive for L. interrogans serovar bratislava and for vesicular stomatitis virus, New Jersey strain. No samples were positive for ADV or Brucella. The seroprevalence of antibodies against L. interrogans was similar to that observed in other studies. Icterohaemorrhagiae appears to be a common serovar among in situ and ex situ peccary populations. Positive antibodies against PVC-2 represent a novel report of exposure to this pathogen in Colombian peccaries. Our results indicate the possible transmission of various pathogens, important for pig farms, in the studied pig and peccaries.

  12. Development of a new LAMP assay for the detection of CSFV strains from Cuba: a proof-of-concept study.

    PubMed

    Postel, Alexander; Pérez, Lester J; Perera, Carmen L; Schmeiser, Stefanie; Meyer, Denise; Meindl-Boehmer, Alexandra; Rios, Liliam; Austermann-Busch, Sophia; Frias-Lepoureau, Maria T; Becher, Paul

    2015-06-01

    Classical swine fever (CSF) is a devastating animal disease of great economic impact worldwide. In many countries, CSF has been endemic for decades, and vaccination of domestic pigs is one of the measures to control the disease. Consequently, differentiating infected from vaccinated animals by antibody ELISA screening is not applicable. In some countries, such as Cuba, lack of molecular techniques for sensitive, rapid and reliable detection of virus genomes is a critical point. To overcome this problem, an easy-to-use one-tube assay based on the loop-mediated isothermal amplification (LAMP) principle has been developed for detection of the genome of CSF virus (CSFV) of endemic Cuban genotype 1.4 isolates. The assay reliably detected recent isolates from three different regions of Cuba with an analytical sensitivity 10-100 times lower than that of quantitative reverse transcription RT-qPCR. Diagnostic test sensitivity was examined using reference sera from two groups of pigs experimentally infected with Cuban virulent strain CSF0705 "Margarita" and the recent field isolate CSF1058 "Pinar del Rio". Differences in pathogenicity of the two viruses were reflected in the clinical course of disease as well as in virus loads of blood samples. Low viral RNA loads in samples from pigs infected with the field isolate caused serious detection problems in RT-LAMP as well as in RT-qPCR. Thus, it will be necessary in future research to focus on targeted sampling of diseased animals and to restrict diagnosis to the herd level in order to establish LAMP as an efficient tool for diagnosing CSF under field conditions.

  13. Evaluation of the risk of classical swine fever (CSF) spread from backyard pigs to other domestic pigs by using the spatial stochastic disease spread model Be-FAST: the example of Bulgaria.

    PubMed

    Martínez-López, Beatriz; Ivorra, Benjamin; Ramos, Angel Manuel; Fernández-Carrión, Eduardo; Alexandrov, Tsviatko; Sánchez-Vizcaíno, José Manuel

    2013-07-26

    The study presented here is one of the very first aimed at exploring the potential spread of classical swine fever (CSF) from backyard pigs to other domestic pigs. Specifically, we used a spatial stochastic spread model, called Be-FAST, to evaluate the potential spread of CSF virus (CSFV) in Bulgaria, which holds a large number of backyards (96% of the total number of pig farms) and is one of the very few countries for which backyard pigs and farm counts are available. The model revealed that, despite backyard pigs being very likely to become infected, infections from backyard pigs to other domestic pigs were rare. In general, the magnitude and duration of the CSF simulated epidemics were small, with a median [95% PI] number of infected farms per epidemic of 1 [1,4] and a median [95% PI] duration of the epidemic of 44 [17,101] days. CSFV transmission occurs primarily (81.16%) due to indirect contacts (i.e. vehicles, people and local spread) whereas detection of infected premises was mainly (69%) associated with the observation of clinical signs on farm rather than with implementation of tracing or zoning. Methods and results of this study may support the implementation of risk-based strategies more cost-effectively to prevent, control and, ultimately, eradicate CSF from Bulgaria. The model may also be easily adapted to other countries in which the backyard system is predominant. It can also be used to simulate other similar diseases such as African swine fever. Copyright © 2013 Elsevier B.V. All rights reserved.

  14. The effects of modified constraint-induced movement therapy and mirror therapy on upper extremity function and its influence on activities of daily living.

    PubMed

    Ju, Yumi; Yoon, In-Jin

    2018-01-01

    [Purpose] Modified constraint-induced movement therapy and mirror therapy are recognized as stroke rehabilitation methods. The aim of the present study was to determine whether these therapies influence upper extremity function and whether upper extremity function influences the ability to perform activities of daily living in further. [Subjects and Methods] Twenty-eight stroke patients participated in the study. Interventions were administered five times per week for 3 weeks. Activities of daily living or self-exercise were performed after modified constraint-induced movement therapy or mirror therapy, respectively. Analyses were performed on the results of the Manual Function Test and the Korean version of the Modified Barthel Index to determine the factors influencing activities of daily living. [Results] Both groups showed improvement in upper extremity function, but only the modified constraint-induced movement therapy group showed a correlation between upper extremity function and performance in the hygiene, eating, and dressing. The improved hand manipulation function found in the modified constraint-induced movement therapy had statistically significant influences on eating and dressing. [Conclusion] Our results suggest that a patient's attempts to move the affected side result in improved performance in activities of daily living as well as physical function.

  15. The effects of modified constraint-induced movement therapy and mirror therapy on upper extremity function and its influence on activities of daily living

    PubMed Central

    Ju, Yumi; Yoon, In-Jin

    2018-01-01

    [Purpose] Modified constraint-induced movement therapy and mirror therapy are recognized as stroke rehabilitation methods. The aim of the present study was to determine whether these therapies influence upper extremity function and whether upper extremity function influences the ability to perform activities of daily living in further. [Subjects and Methods] Twenty-eight stroke patients participated in the study. Interventions were administered five times per week for 3 weeks. Activities of daily living or self-exercise were performed after modified constraint-induced movement therapy or mirror therapy, respectively. Analyses were performed on the results of the Manual Function Test and the Korean version of the Modified Barthel Index to determine the factors influencing activities of daily living. [Results] Both groups showed improvement in upper extremity function, but only the modified constraint-induced movement therapy group showed a correlation between upper extremity function and performance in the hygiene, eating, and dressing. The improved hand manipulation function found in the modified constraint-induced movement therapy had statistically significant influences on eating and dressing. [Conclusion] Our results suggest that a patient’s attempts to move the affected side result in improved performance in activities of daily living as well as physical function. PMID:29410571

  16. Evaluation of the efficacy of a new modified live porcine reproductive and respiratory syndrome virus (PRRSV) vaccine (Fostera PRRS) against heterologous PRRSV challenge.

    PubMed

    Park, Changhoon; Seo, Hwi Won; Han, Kiwon; Kang, Ikjae; Chae, Chanhee

    2014-08-27

    The objective of this study was to evaluate a new modified live porcine reproductive and respiratory syndrome virus (PRRSV) vaccine (Fostera PRRS, Zoetis, Florham, NJ, USA) that was based on a virulent US PRRSV isolate (P129) attenuated using CD163-expressing cell lines. Sixty-four PRRSV-seronegative 3-week-old pigs were randomly divided into the following four groups: vaccinated challenged (group 1), vaccinated unchallenged (group 2), unvaccinated challenged (group 3), and unvaccinated unchallenged (group 4). The pigs in groups 1 and 2 were immunized with a 2.0 mL dose of modified live PRRSV vaccine at 21 days of age, according to the manufacturer's recommendations. At 56 days of age (0 days post-challenge), the pigs in groups 1 and 3 were inoculated intranasally with 3 mL of tissue culture fluid containing 10(5) 50% tissue culture infective dose (TCID50)/mL of PRRSV (SNUVR090851 strain, fourth passage in MARC-145 cells). Vaccinated challenged pigs exhibited significantly lower (P<0.05) respiratory scores, viremia, macroscopic and microscopic lung lesion scores, and PRRSV-antigen with interstitial pneumonia than unvaccinated challenged pigs. The induction of PRRSV-specific IFN-γ-SCs by the new modified live PRRSV vaccine produced a protective immune response, leading to the reduction of PRRSV viremia. Although the new modified live PRRSV vaccine is not effective against heterologous PRRSV challenge, the new modified live PRRSV vaccine was able to reduce the levels of viremia and nasal shedding, and severity of PRRSV-induced lesions after challenging virus under experimental conditions. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Modified constraint-induced movement therapy for clients with chronic stroke: interrupted time series (ITS) design.

    PubMed

    Park, JuHyung; Lee, NaYun; Cho, YongHo; Yang, YeongAe

    2015-03-01

    [Purpose] The purpose of this study was to investigate the impact that modified constraint-induced movement therapy has on upper extremity function and the daily life of chronic stroke patients. [Subjects and Methods] Modified constraint-induced movement therapy was conduct for 2 stroke patients with hemiplegia. It was performed 5 days a week for 2 weeks, and the participants performed their daily living activities wearing mittens for 6 hours a day, including the 2 hours of the therapy program. The assessment was conducted 5 times in 3 weeks before and after intervention. The upper extremity function was measured using the box and block test and a dynamometer, and performance daily of living activities was assessed using the modified Barthel index. The results were analyzed using a scatterplot and linear regression. [Results] All the upper extremity functions of the participants all improved after the modified constraint-induced movement therapy. Performance of daily living activities by participant 1 showed no change, but the results of participant 2 had improved after the intervention. [Conclusion] Through the results of this research, it was identified that modified constraint-induced movement therapy is effective at improving the upper extremity functions and the performance of daily living activities of chronic stroke patients.

  18. 9 CFR 112.7 - Special additional requirements.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... a biological product containing inactivated rabies virus, carton labels, enclosures, and all but... biological product containing modified live rabies virus, the carton labels, enclosures, and all but very... Any Other Animal!” (2) For other vaccines containing modified live rabies virus, the statement “For...

  19. Response of gray foxes to modified live-virus canine distemper vaccines.

    PubMed

    Halbrooks, R D; Swango, L J; Schnurrenberger, P R; Mitchell, F E; Hill, E P

    1981-12-01

    Ten gray foxes seronegative for canine distemper virus were vaccinated with 1 of 3 commercial modified live-virus canine distemper vaccines. Of 5 foxes receiving vaccine A (chicken tissue culture origin), 4 developed significant titers (greater than or equal to 1:100) of neutralizing antibody to canine distemper virus and remained clinically normal after vaccination. Two of 3 foxes vaccinated with vaccine B (canine cell line origin) and both foxes receiving vaccine C (canine cell line origin) died of vaccine-induced distemper. Five unvaccinated control foxes died of distemper after a known occasion for contact transmission of virus from a fox vaccinated with vaccine B. The results suggested that the chicken tissue culture origin modified live-virus canine distemper vaccine is probably safe for normal adult gray foxes, whereas the canine cell origin vaccines are hazardous. The results of this study tended to corroborate anecdotal experiences of veterinarians who have observed that gray foxes frequently die from distemper soon after vaccination with modified live-virus canine distemper vaccines.

  20. Feline panleukopenia virus, feline herpesvirus-1 and feline calicivirus antibody responses in seronegative specific pathogen-free kittens after parenteral administration of an inactivated FVRCP vaccine or a modified live FVRCP vaccine.

    PubMed

    Lappin, Michael R

    2012-02-01

    Two groups of feline panleukopenia (FPV), feline calicivirus (FCV) and feline herpesvirus 1 (FHV-1) seronegative kittens (six cats per group) were administered one of two feline viral rhinotracheitis, calcivirus and panleukopenia (FVRCP) vaccines subcutaneously (one inactivated and one modified live) and the serological responses to each agent were followed over 49 days (days 0, 2, 5, 7, 10, 14, 21, 28, 35, 42, 49). While the kittens administered the modified live FPV vaccine were more likely to seroconvert on day 7 after the first inoculation than kittens administered the inactivated vaccine, all kittens had seroconverted by day 14. In contrast, FHV-1 serological responses were more rapid following administration of the inactivated FVRCP vaccine when compared with the modified live FVRCP vaccine. There were no statistical differences between the serological response rates between the two FVRCP vaccines in regard to FCV.

  1. Mammalian Toxicological Evaluations of TNT Wastewaters. Volume III. Acute and Subacute Mammalian Toxicity of Condensate Water.

    DTIC Science & Technology

    1979-04-01

    Dellen Labs., Inc., Omaha, Nebr- 68134 I The Distemper and Hepatitis fractions are a modified live virus with a canine tissue culture origin. The...Leptospirosis fraction contains physically inactivated Leptospira Canicola and Icterohemorrhagiae Bacterin. DISTEMPER Modified Live Virus-Chick Tissue...ISTEMER-HEPATITIS-LEPTOSPI ROS IS DELCINE HL produced by Del len Labs., Inc., Omaha, Nebraska 68134 The Distemper and Hepatitis fractions are a modified

  2. Development of a broadly protective modified-live virus vaccine candidate against porcine reproductive and respiratory syndrome virus

    USDA-ARS?s Scientific Manuscript database

    Modified-live virus (MLV) vaccines are widely used to protect pigs against porcine reproductive and respiratory syndrome virus (PRRSV). However, current MLV vaccines do not confer adequate levels of heterologous protection, presumably due to the substantial genetic diversity of PRRSV isolates circul...

  3. [Serological detection of emerging viral infections in wild boars from different hunting regions of Southern Germany].

    PubMed

    Sattler, T; Sailer, E; Wodak, E; Schmoll, F

    2012-01-01

    Wild boars represent a possible virus reservoir for notifiable diseases of farm animals, including Aujeszky's disease (AD) and classical swine fever (CSF). Monitoring of the epidemiological situation in the wild boar population is especially relevant in countries that are officially free from these diseases. Apart from OIE-notifiable diseases, other viral agents that are widely distributed and play a significant role in farm animals, such as the porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type2 (PCV-2), and swine influenza virus (SIV), are sporadically detected in wild boars. Thus, the wild boar population is a potential source for maintenance of these infections in farm animals. The aim of this study was therefore to test for antibodies to the indicated emerging viral infections in wild boars in several hunting regions of Southern Germany. Blood serum of 94 shot wild boars from 19 hunting regions in Bavaria, Rhineland-Palatinate and Baden-Württemberg were collected. Antibodies to AD virus (ADV), CSF virus (CSFV), PRRSV, SIV (H1N1) (all by IDEXX ELISA) and PCV-2 (IgM and IgG by Ingenasa ELISA) in blood serum were determined. Antibodies to ADV were detected in four animals (4.2%), to PRRSV in one animal (1.2%), to SIV (H1N1) in two animals (2.1%) and to PCV-2 (IgG) in 15 animals (16.0%) of which three animals also had antibodies to PCV-2 (IgM) (3.2%). CSFV antibodies were not detected in the examined wild boars. Compared to other studies in several European and American states, the seroprevalence to the tested emerging diseases was low in this study. Nevertheless, the wild boar population may still be a virus reservoir and therefore a source of infection for domestic pigs. This is especially important in the case of notifiable diseases, like AD and CSF. Therefore, a continuous monitoring of those diseases in the wild boar population would be advisable.

  4. Novel poly-uridine insertion in the 3'UTR and E2 amino acid substitutions in a low virulent classical swine fever virus.

    PubMed

    Coronado, Liani; Liniger, Matthias; Muñoz-González, Sara; Postel, Alexander; Pérez, Lester Josue; Pérez-Simó, Marta; Perera, Carmen Laura; Frías-Lepoureau, Maria Teresa; Rosell, Rosa; Grundhoff, Adam; Indenbirken, Daniela; Alawi, Malik; Fischer, Nicole; Becher, Paul; Ruggli, Nicolas; Ganges, Llilianne

    2017-03-01

    In this study, we compared the virulence in weaner pigs of the Pinar del Rio isolate and the virulent Margarita strain. The latter caused the Cuban classical swine fever (CSF) outbreak of 1993. Our results showed that the Pinar del Rio virus isolated during an endemic phase is clearly of low virulence. We analysed the complete nucleotide sequence of the Pinar del Rio virus isolated after persistence in newborn piglets, as well as the genome sequence of the inoculum. The consensus genome sequence of the Pinar del Rio virus remained completely unchanged after 28days of persistent infection in swine. More importantly, a unique poly-uridine tract was discovered in the 3'UTR of the Pinar del Rio virus, which was not found in the Margarita virus or any other known CSFV sequences. Based on RNA secondary structure prediction, the poly-uridine tract results in a long single-stranded intervening sequence (SS) between the stem-loops I and II of the 3'UTR, without major changes in the stem- loop structures when compared to the Margarita virus. The possible implications of this novel insertion on persistence and attenuation remain to be investigated. In addition, comparison of the amino acid sequence of the viral proteins E rns , E1, E2 and p7 of the Margarita and Pinar del Rio viruses showed that all non-conservative amino acid substitutions acquired by the Pinar del Rio isolate clustered in E2, with two of them being located within the B/C domain. Immunisation and cross-neutralisation experiments in pigs and rabbits suggest differences between these two viruses, which may be attributable to the amino acid differences observed in E2. Altogether, these data provide fresh insights into viral molecular features which might be associated with the attenuation and adaptation of CSFV for persistence in the field. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Valuing lives and allocating resources: a defense of the modified youngest first principle of scarce resource distribution.

    PubMed

    Tallman, Ruth

    2014-06-01

    In this paper, I argue that the 'modified youngest first' principle provides a morally appropriate criterion for making decisions regarding the distribution of scarce medical resources, and that it is morally preferable to the simple 'youngest first' principle. Based on the complete lives system's goal of maximizing complete lives rather than individual life episodes, I argue that essential to the value we see in complete lives is the first person value attributed by the experiencer of that life. For a life to be 'complete' or 'incomplete,' the subject of that life must be able to understand the concept of a complete life, to have started goals and projects, and to know what it would be for that life to be complete. As the very young are not able to do this, it can reasonably be said that their characteristically human lives have not yet begun, giving those accepting a complete lives approach good reason to accept the modified youngest first principle over a simple 'youngest first' approach. © 2012 John Wiley & Sons Ltd.

  6. Thrombocytopenia associated with vaccination of a dog with a modified-live paramyxovirus vaccine.

    PubMed

    McAnulty, J F; Rudd, R G

    1985-06-01

    Thrombocytopenia (10,000/mm3), with hematochezia and melena, appeared in a dog 8 days after it was given modified-live canine distemper, virus vaccine and persisted for approximately 5 days. Clinical investigation discounted other possible causes of thrombocytopenia; the condition was considered to be associated with vaccination. The problem spontaneously resolved. The appearance of thrombocytopenia after modified-live canine distemper virus vaccination is not unknown and may assume a severe form. This condition may be mistaken for idiopathic thrombocytopenia of immune origin, and in other instances, it may contribute significantly to surgical risk if concurrent coagulation disorders are present. Administration of levamisole HCl may alleviate the decrease in platelet count in affected animals.

  7. A new modified live porcine reproductive and respiratory syndrome vaccine improves growth performance in pigs under field conditions.

    PubMed

    Park, Changhoon; Seo, Hwi Won; Kang, Ikjae; Jeong, Jiwoon; Choi, Kyuhyung; Chae, Chanhee

    2014-09-01

    The change in growth performance resulting from a new modified live porcine reproductive and respiratory syndrome (PRRS) vaccine was evaluated under field conditions for registration with the government as guided by the Republic of Korea's Animal and Plant Quarantine Agency. Three farms were selected based on their history of PRRS-associated respiratory diseases. On each farm, a total of 45 3-week-old pigs were randomly allocated to one of two treatment groups, (i) vaccinated (n = 25) or (ii) control (n = 20) animals. A new modified live PRRSV vaccine increased market weight by 1.26 kg/pig (104.71 kg versus 103.45 kg; P < 0.05) and decreased mortality by 17% (1.33% versus 18.33%; P < 0.05). Pathological examination indicated that vaccination effectively reduced microscopic lung lesions compared with control animals on the 3 farms. Thus, the new modified live PRRS vaccine improved growth performance and decreased mortality and lung lesions when evaluated under field conditions. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  8. Modifying your Physics and Astronomy Courses to Incorporate Heliophysics - Some Examples

    NASA Astrophysics Data System (ADS)

    Cebulka, Rebecca; Cox, Amanda; Rodriguez Garrigues, Alvar; Hoshino, Laura; Fitzgerald, Cullen; Montgomery, M.; Al-Rawi, Ahlam N.; Velissaris, Christos; Flitsiyan, Elena

    2016-01-01

    Although physics and astronomy courses include heliophysics topics, students still leave the courses without knowing what heliophysics is and how heliophysics relates to their daily lives. To meet goals of NASA's Living With a Star Program, UCF Physics has modified courses such as SCALE-UP: Electricity and Magnetism for Engineers and Scientists, Astronomy (for non-science majors), and Astrophysics to include heliophysics topics. In this poster, we present the previous labs, the student-modified labs to incorporate heliophysics, and we present student learning statistics.

  9. MontanideTM Gel01 ST adjuvant enhances PRRS modified live vaccine efficacy by regulating porcine humoral and cellular immune responses

    USDA-ARS?s Scientific Manuscript database

    Porcine reproductive and respiratory syndrome (PRRS) is a devastating disease caused by the PRRS virus. The MontanideTM class of flexible polymeric adjuvants has recently been shown to enhance protective immunity against PRRSV infection in piglets when used in combination with PRRS modified live vac...

  10. Adenovirus 2, Bordetella bronchiseptica, and Parainfluenza Molecular Diagnostic Assay Results in Puppies After vaccination with Modified Live Vaccines.

    PubMed

    Ruch-Gallie, R; Moroff, S; Lappin, M R

    2016-01-01

    Canine adenovirus 2, parainfluenza, and Bordetella bronchiseptica cause respiratory disease in dogs, and each has a modified live intranasal vaccine available. Molecular diagnostic assays to amplify specific nucleic acids are available for each of these agents. If positive molecular diagnostic assay results are common after vaccination, the positive predictive value of the diagnostic assays for disease would be decreased. To determine the impact of administration of commercially available modified live topical adenovirus 2, B. bronchiseptica, and parainfluenza vaccine has on the results of a commercially available PCR panel. Eight puppies from a research breeding facility negative for these pathogens. Blinded prospective pilot study. Puppies were vaccinated with a single dose of modified live topical adenovirus 2, B. bronchiseptica, and parainfluenza and parenteral dose of adenovirus 2, canine distemper virus, and parvovirus. Nasal and pharyngeal swabs were collected on multiple days and submitted for PCR assay. Nucleic acids of all 3 organisms contained in the topical vaccine were detected from both samples multiple times through 28 days after vaccination with higher numbers of positive samples detected between days 3 and 10 after vaccination. Vaccine status should be considered when interpreting respiratory agent PCR results if modified live vaccines have been used. Development of quantitative PCR and wild-type sequencing are necessary to improve positive predictive value of these assays by distinguishing vaccinate from natural infection. Copyright © 2015 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

  11. Clinical trials with canine distemper vaccines in exotic carnivores.

    PubMed

    Montali, R J; Bartz, C R; Teare, J A; Allen, J T; Appel, M J; Bush, M

    1983-12-01

    Two types of killed canine distemper virus (CDV) vaccine and a modified-live CDV vaccine were clinically evaluated in four species of exotic carnivores. In 16 trials in which 13 red pandas (Ailurus fulgens) were given the killed vaccine, only 1 animal had a virus-neutralization titer that exceeded 1:100. A red panda given modified-live CDV vaccine deemed safe for gray foxes and ferrets died of bacterial pneumonia 16 days later. There was no pathologic evidence of canine distemper in that panda. The same modified-live vaccine proved to be immunogenic and safe in 12 bush dogs (Speothos venaticus), 5 maned wolves (Chrysocyon brachyurus), and 3 fennec foxes (Fennecus zerda) in which virus-neutralization titers often exceeded 1:512 and persisted for several months after vaccination.

  12. A modified live canine parvovirus strain with novel plaque characteristics. I. Viral attenuation and dog response.

    PubMed

    Carmichael, L E; Joubert, J C; Pollock, R V

    1981-10-01

    A canine parvovirus (CPV) strain (C-780916) was found attenuated for pups at 80, but not after 51 serial passages in dog kidney cell (DKC) cultures. A variant viral population ('large plaque') emerged after prolonged cultivation in DKC cultures that may be associated with reduced native virulence. Dogs vaccinated with modified CPV developed high hemagglutination-inhibiting (HI) antibody titers within 4 days of incoluation and antibody persisted. Vaccinated animals shed small amounts of virus in the feces that spread to contact dogs. After five back-passages in dogs the modified strain was not pathogenic for pups and the plaque characteristics of the virus isolated from the feces were typical of the attenuated strain. The modified live CPV did not cause infection of the fetus when inoculated parenterally into pregnant bitches at various stages of gestation. It was not pathogenic for neonatal pups. These results suggest that a safe and effective live homologous (CPV) vaccine has been developed which should aid substantially in controlling CPV infection.

  13. Potency control of modified live viral vaccines for veterinary use.

    PubMed

    Terpstra, C; Kroese, A H

    1996-04-01

    This paper reviews various aspects of efficacy, and methods for assaying the potency of modified live viral vaccines. The pros and cons of parametric versus non-parametric methods for analysis of potency assays are discussed and critical levels of protection, as determined by the target(s) of vaccination, are exemplified. Recommendations are presented for designing potency assays on master virus seeds and vaccine batches.

  14. Potency control of modified live viral vaccines for veterinary use.

    PubMed

    Terpstra, C; Kroese, A H

    1996-01-01

    This paper reviews various aspects of efficacy, and methods for assaying the potency of modified live viral vaccines. The pros and cons of parametric versus non-parametric methods for analysis of potency assays are discussed and critical levels of protection, as determined by the target(s) of vaccination, are exemplified. Recommendations are presented for designing potency assays on master virus seeds and vaccine batches.

  15. Immunogenicity of an intranasally administered modified live canine parvovirus type 2b vaccine in pups with maternally derived antibodies.

    PubMed

    Martella, Vito; Cavalli, Alessandra; Decaro, Nicola; Elia, Gabriella; Desario, Costantina; Campolo, Marco; Bozzo, Giancarlo; Tarsitano, Elvira; Buonavoglia, Canio

    2005-10-01

    The ability of a modified live canine parvovirus type 2b vaccine to elicit active immunization in pups with maternally derived antibodies (MDA) by intranasal administration was evaluated. The vaccine induced seroconversion in 100% of pups with MDA titers of < or = 80 and in 51.6% of pups with titers between 160 and 320.

  16. Immunogenicity of an Intranasally Administered Modified Live Canine Parvovirus Type 2b Vaccine in Pups with Maternally Derived Antibodies

    PubMed Central

    Martella, Vito; Cavalli, Alessandra; Decaro, Nicola; Elia, Gabriella; Desario, Costantina; Campolo, Marco; Bozzo, Giancarlo; Tarsitano, Elvira; Buonavoglia, Canio

    2005-01-01

    The ability of a modified live canine parvovirus type 2b vaccine to elicit active immunization in pups with maternally derived antibodies (MDA) by intranasal administration was evaluated. The vaccine induced seroconversion in 100% of pups with MDA titers of ≤80 and in 51.6% of pups with titers between 160 and 320. PMID:16210491

  17. Evaluation of reproductive protection against bovine viral diarrhea virus and bovine herpesvirus-1 afforded by annual revaccination with modified-live viral or combination modified-live/killed viral vaccines after primary vaccination with modified-live viral vaccine.

    PubMed

    Walz, Paul H; Givens, M Daniel; Rodning, Soren P; Riddell, Kay P; Brodersen, Bruce W; Scruggs, Daniel; Short, Thomas; Grotelueschen, Dale

    2017-02-15

    The objective of this study was to compare reproductive protection in cattle against bovine viral diarrhea virus (BVDV) and bovine herpesvirus 1 (BoHV-1) provided by annual revaccination with multivalent modified-live viral (MLV) vaccine or multivalent combination viral (CV) vaccine containing temperature-sensitive modified-live BoHV-1 and killed BVDV when MLV vaccines were given pre-breeding to nulliparous heifers. Seventy-five beef heifers were allocated into treatment groups A (n=30; two MLV doses pre-breeding, annual revaccination with MLV vaccine), B (n=30; two MLV doses pre-breeding, annual revaccination with CV vaccine) and C (n=15; saline in lieu of vaccine). Heifers were administered treatments on days 0 (weaning), 183 (pre-breeding), 366 (first gestation), and 738 (second gestation). After first calving, primiparous cows were bred, with pregnancy assessment on day 715. At that time, 24 group A heifers (23 pregnancies), 23 group B heifers (22 pregnancies), and 15 group C heifers (15 pregnancies) were commingled with six persistently infected (PI) cattle for 16days. Ninety-nine days after PI removal, cows were intravenously inoculated with BoHV-1. All fetuses and live offspring were assessed for BVDV and BoHV-1. Abortions occurred in 3/23 group A cows, 1/22 group B cows, and 11/15 group C cows. Fetal infection with BVDV or BoHV-1 occurred in 4/23 group A offspring, 0/22 group B offspring, and 15/15 group C offspring. This research demonstrates efficacy of administering two pre-breeding doses of MLV vaccine with annual revaccination using CV vaccine to prevent fetal loss due to exposure to BVDV and BoHV-1. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  18. The possible enhancement of parvovirus vaccination on the mortality rate of diseased dogs.

    PubMed

    Brenner, J; Markus, R; Klopfer-Orgad, U; Trainin, Z

    1989-09-01

    A survey was conducted by using questionnaires dealing with morbidity and mortality of young dogs. Seventy-one clinical evaluations of diseased dogs were included in this survey. The cases were divided into 3 groups according to their vaccination history: 1. dogs vaccinated with the parvovirus and distemper live modified vaccine; 2. dogs vaccinated with the killed parvovirus vaccine; 3. non-vaccinated dogs. Statistical analysis of the information obtained by analyzing the questionnaires gave the following results: The survival rate of the non-vaccinated and killed parvovirus vaccine-vaccinated dogs was significantly higher than that of the dogs which were vaccinated by the live modified vaccine. No such difference was encountered between groups 2 and 3. This finding, once more, supports the notion that the live modified parvovirus vaccine may have an immunosuppressive property.

  19. The effect of reagents mimicking oxidative stress on fibrinogen function.

    PubMed

    Štikarová, Jana; Kotlín, Roman; Riedel, Tomáš; Suttnar, Jiří; Pimková, Kristýna; Chrastinová, Leona; Dyr, Jan E

    2013-01-01

    Fibrinogen is one of the plasma proteins most susceptible to oxidative modification. It has been suggested that modification of fibrinogen may cause thrombotic/bleeding complications associated with many pathophysiological states of organism. We exposed fibrinogen molecules to three different modification reagents-malondialdehyde, sodium hypochlorite, and peroxynitrite-that are presented to various degrees in different stages of oxidative stress. We studied the changes in fibrin network formation and platelet interactions with modified fibrinogens under flow conditions. The fastest modification of fibrinogen was caused by hypochlorite. Fibers from fibrinogen modified with either reagent were thinner in comparison with control fibers. We found that platelet dynamic adhesion was significantly lower on fibrinogen modified with malondialdehyde and significantly higher on fibrinogen modified either with hypochlorite or peroxynitrite reflecting different prothrombotic/antithrombotic properties of oxidatively modified fibrinogens. It seems that, in the complex reactions ongoing in living organisms at conditions of oxidation stress, hypochlorite modifies proteins (e.g., fibrinogen) faster and more preferentially than malondialdehyde. It suggests that the prothrombotic effects of prior fibrinogen modifications may outweigh the antithrombotic effect of malondialdehyde-modified fibrinogen in real living systems.

  20. The Effect of Reagents Mimicking Oxidative Stress on Fibrinogen Function

    PubMed Central

    Štikarová, Jana; Kotlín, Roman; Riedel, Tomáš; Suttnar, Jiří; Pimková, Kristýna; Chrastinová, Leona; Dyr, Jan E.

    2013-01-01

    Fibrinogen is one of the plasma proteins most susceptible to oxidative modification. It has been suggested that modification of fibrinogen may cause thrombotic/bleeding complications associated with many pathophysiological states of organism. We exposed fibrinogen molecules to three different modification reagents—malondialdehyde, sodium hypochlorite, and peroxynitrite—that are presented to various degrees in different stages of oxidative stress. We studied the changes in fibrin network formation and platelet interactions with modified fibrinogens under flow conditions. The fastest modification of fibrinogen was caused by hypochlorite. Fibers from fibrinogen modified with either reagent were thinner in comparison with control fibers. We found that platelet dynamic adhesion was significantly lower on fibrinogen modified with malondialdehyde and significantly higher on fibrinogen modified either with hypochlorite or peroxynitrite reflecting different prothrombotic/antithrombotic properties of oxidatively modified fibrinogens. It seems that, in the complex reactions ongoing in living organisms at conditions of oxidation stress, hypochlorite modifies proteins (e.g., fibrinogen) faster and more preferentially than malondialdehyde. It suggests that the prothrombotic effects of prior fibrinogen modifications may outweigh the antithrombotic effect of malondialdehyde-modified fibrinogen in real living systems. PMID:24235886

  1. Expected Net Benefit of Vaccinating Rangeland Sheep against Bluetongue Virus Using a Modified-Live versus Killed Virus Vaccine

    PubMed Central

    Munsick, Tristram R.; Peck, Dannele E.; Ritten, John P.; Jones, Randall; Jones, Michelle; Miller, Myrna M.

    2017-01-01

    Recurring outbreaks of bluetongue virus in domestic sheep of the US Intermountain West have prompted questions about the economic benefits and costs of vaccinating individual flocks against bluetongue (BT) disease. We estimate the cost of a BT outbreak on a representative rangeland sheep operation in the Big Horn Basin of the state of Wyoming using enterprise budgets and stochastic simulation. The latter accounts for variability in disease severity and lamb price, as well as uncertainty about when an outbreak will occur. We then estimate the cost of purchasing and administering a BT vaccine. Finally, we calculate expected annual net benefit of vaccinating under various outbreak intervals. Expected annual net benefit is calculated for both a killed virus (KV) vaccine and modified-live virus vaccine, using an observed price of $0.32 per dose for modified-live and an estimated price of $1.20 per dose for KV. The modified-live vaccine’s expected annual net benefit has a 100% chance of being positive for an outbreak interval of 5, 10, or 20 years, and a 77% chance of being positive for a 50-year interval. The KV vaccine’s expected annual net benefit has a 97% chance of being positive for a 5-year outbreak interval, and a 42% chance of being positive for a 10-year interval. A KV vaccine is, therefore, unlikely to be economically attractive to producers in areas exposed less frequently to BT disease. A modified-live vaccine, however, requires rigorous authorization before legal use can occur in Wyoming. To date, no company has requested to manufacture a modified-live vaccine for commercial use in Wyoming. The KV vaccine poses less risk to sheep reproduction and less risk of unintentional spread, both of which facilitate approval for commercial production. Yet, our results show an economically consequential tradeoff between a KV vaccine’s relative safety and higher cost. Unless the purchase price is reduced below our assumed $1.20 per dose, producer adoption of a KV vaccine for BT is likely to be low in the study area. This tradeoff between cost and safety should be considered when policymakers regulate commercial use of the two vaccine types. PMID:29075635

  2. Combined administration in a single injection of a feline multivalent modified live vaccine against FHV, FCV, and FPLV together with a recombinant FeLV vaccine is both safe and efficacious for all four major feline viral pathogens.

    PubMed

    Kanellos, Theo; Sutton, David J; Salisbury, Claire F; Chalmers, William Stuart K

    2008-08-01

    Nobivac Tricat, a lyophilised trivalent modified live attenuated vaccine is routinely used to protect cats against three commonly diagnosed feline viral pathogens namely herpesvirus, calicivirus and panleukopenia virus. The recognition of feline leukaemia virus (FeLV) as an important viral pathogen has prompted the development of an efficacious liquid recombinant subunit FeLV vaccine (p45 envelope protein). Lyophilised Tricat vaccine was dissolved in the liquid FeLV vaccine and no detectable deleterious effect on the titre of any of the live virus components was observed after 2h incubation. In vivo studies where the vaccines were mixed in the same syringe prior to inoculation showed no alteration to the safety profile assessed by repeat and overdose studies. Serological comparisons of the modified live viral antibody titres showed no evidence of reduced responses following administration of the mixed products. Challenge studies using pathogenic herpesvirus and FeLV revealed no difference in the degree of clinical protection. This paper shows that neither safety nor efficacy is adversely affected as a result of mixing the two vaccines.

  3. Validity and reliability of the modified Chinese version of the Older People's Quality of Life Questionnaire (OPQOL) in older people living alone in China.

    PubMed

    Chen, Yu; Hicks, Allan; While, Alison E

    2014-12-01

    This study aimed to test the validity and reliability of a modified Chinese version of the OPQOL among older people living alone in China. China has an ageing population with an increasing number of older people living alone who may have a poorer quality of life (QoL) in the light of the traditional culture of collectivism and filial piety. An appropriate instrument is important to assess their QoL. The Older People's Quality of Life Questionnaire (OPQOL) was developed directly from the views of older people and has been validated in England. There has been no psychometric evaluation of the scale in China. The OPQOL was translated and modified prior to being administered to a stratified random cluster sample of 521 older people living alone. Validity was assessed through convergent validity, discriminant validity and construct validity. Reliability was assessed through internal consistency and test-retest reliability. Exploratory factor analysis indicated eight factors accounting for 63.77% of the variance. The convergent validity was supported by moderate correlations with functional ability, social support and loneliness with Spearman's rho of -0.50, 0.49 and -0.53, respectively. The discriminant validity was confirmed by differentiating QoL scores between the depressed and non-depressed groups. The Cronbach's α coefficient was 0.90 for the total scale and over 0.70 for most of its dimensions. The 2-week test-retest reliability ranged from 0.53 to 0.87. The modified Chinese version of the Older People's Quality of Life has acceptable validity and reliability as a useful instrument to measure the QoL of older people living alone in China. © 2013 John Wiley & Sons Ltd.

  4. Use of modified live feline panleukopenia virus vaccine to immunize dogs against canine parvovirus.

    PubMed

    Pollock, R V; Carmichael, L E

    1983-02-01

    Modified live feline panleukopenia virus (FPLV) vaccine protected dogs against canine parvovirus (CPV) infection. However, unlike the long-lived (greater than or equal to 20-month) immunity engendered by CPV infection, the response of dogs to living FPLV was variable. Doses of FPLV (snow leopard strain) in excess of 10(5.7) TCID50 were necessary for uniform immunization; smaller inocula resulted in decreased success rates. The duration of immunity, as measured by the persistence of hemagglutination-inhibiting antibody, was related to the magnitude of the initial response to vaccination; dogs with vigorous initial responses resisted oronasal CPV challenge exposure 6 months after vaccination, and hemagglutination-inhibiting antibodies persisted in such dogs for greater than 1 year. Limited replication of FPLV in dogs was demonstrated, but unlike CPV, the feline virus did not spread to contact dogs or cats. Adverse reactions were not associated with living FPLV vaccination, and FPLV did not interfere with simultaneous response to attenuated canine distemper virus.

  5. Modified blank ammunition injuries.

    PubMed

    Ogunc, Gokhan I; Ozer, M Tahir; Coskun, Kagan; Uzar, Ali Ihsan

    2009-12-15

    Blank firing weapons are designed only for discharging blank ammunition cartridges. Because they are cost-effective, are easily accessible and can be modified to live firearms plus their unclear legal situation in Turkish Law makes them very popular in Turkey. 2004 through 2008, a total of 1115 modified blank weapons were seized in Turkey. Blank firing weapons are easily modified by owners, making them suitable for discharging live firearm ammunition or modified blank ammunitions. Two common methods are used for modification of blank weapons. After the modification, these weapons can discharge the live ammunition. However, due to compositional durability problems with these types of weapons; the main trend is to use the modified blank ammunitions rather than live firearm ammunition fired from modified blank firing weapons. In this study, two types of modified blank weapons and two types of modified blank cartridges were tested on three different target models. Each of the models' shooting side was coated with 1.3+/-2 mm thickness chrome tanned cowhide as a skin simulant. The first model was only coated with skin simulant. The second model was coated with skin simulant and 100% cotton police shirt. The third model was coated with skin simulant and jean denim. After the literature evaluation four high risky anatomic locations (the neck area; the eyes; the thorax area and inguinal area) were pointed out for the steel and lead projectiles are discharged from the modified blank weapons especially in close range (0-50 cm). The target models were designed for these anatomic locations. For the target models six Transparent Ballistic Candle blocks (TCB) were prepared and divided into two test groups. The first group tests were performed with lead projectiles and second group with steel projectile. The shortest penetration depth (lead projectile: 4.358 cm; steel projectile 8.032 cm) was recorded in the skin simulant and jean denim coated block for both groups. In both groups, the longest penetration depth (lead projectile: 6.434 cm; steel projectile 8.608 cm) was recorded in the only skin simulant coated block. And the penetration depth of skin simulant and 100% cotton police shirt coated model was 5.870 cm for lead projectile; 8.440 cm for steel projectile. According to penetration results, national and international legislations and production standards should be re-evaluated in order to prevent the modification of blank weapons and ammunitions. There are three methods for preventing modification of blank weapons: completely closed barrel structure; intersected restrain pieces application; eccentric barrel structure.

  6. Prospective Comparison of Live Evaluation and Video Review in the Evaluation of Operator Performance in a Pediatric Emergency Airway Simulation

    PubMed Central

    House, Joseph B.; Dooley-Hash, Suzanne; Kowalenko, Terry; Sikavitsas, Athina; Seeyave, Desiree M.; Younger, John G.; Hamstra, Stanley J.; Nypaver, Michele M.

    2012-01-01

    Introduction Real-time assessment of operator performance during procedural simulation is a common practice that requires undivided attention by 1 or more reviewers, potentially over many repetitions of the same case. Objective To determine whether reviewers display better interrater agreement of procedural competency when observing recorded, rather than live, performance; and to develop an assessment tool for pediatric rapid sequence intubation (pRSI). Methods A framework of a previously established Objective Structured Assessment of Technical Skills (OSATS) tool was modified for pRSI. Emergency medicine residents (postgraduate year 1–4) were prospectively enrolled in a pRSI simulation scenario and evaluated by 2 live raters using the modified tool. Sessions were videotaped and reviewed by the same raters at least 4 months later. Raters were blinded to their initial rating. Interrater agreement was determined by using the Krippendorff generalized concordance method. Results Overall interrater agreement for live review was 0.75 (95% confidence interval [CI], 0.72–0.78) and for video was 0.79 (95% CI, 0.73–0.82). Live review was significantly superior to video review in only 1 of the OSATS domains (Preparation) and was equivalent in the other domains. Intrarater agreement between the live and video evaluation was very good, greater than 0.75 for all raters, with a mean of 0.81 (95% CI, 0.76–0.85). Conclusion The modified OSATS assessment tool demonstrated some evidence of validity in discriminating among levels of resident experience and high interreviewer reliability. With this tool, intrareviewer reliability was high between live and 4-months' delayed video review of the simulated procedure, which supports feasibility of delayed video review in resident assessment. PMID:23997874

  7. Detection and genotyping of bovine diarrhea virus by reverse transcription-polymerase chain amplification of the 5' untranslated region.

    PubMed

    Letellier, C; Kerkhofs, P; Wellemans, G; Vanopdenbosch, E

    1999-01-01

    A reverse-transcription polymerase chain reaction (RT-PCR) was developed to differentiate the bovine diarrhea virus (BVDV) from other pestiviruses, and to determine the genotype of the BVDV isolates. For this purpose, primer pairs were selected in the 5' untranslated region (5'UTR). The primers BE and B2 were located in highly conserved regions and were pestivirus-specific. Two primer pairs named B3B4 and B5B6 were specific of BVDV genotypes I and II, respectively. With this technique, an amplification product of the expected size was obtained with either the B3B4 or the B5B6 primer pairs for the 107 BVDV isolates tested but not for BDV or CSFV. For some isolates that were grouped in the genotype II, sequence analysis of the PCR fragments confirmed their classification into this genotype.

  8. Three-year rabies duration of immunity in dogs following vaccination with a core combination vaccine against canine distemper virus, canine adenovirus type-1, canine parvovirus, and rabies virus.

    PubMed

    Lakshmanan, Nallakannu; Gore, Thomas C; Duncan, Karen L; Coyne, Michael J; Lum, Melissa A; Sterner, Frank J

    2006-01-01

    Thirty-two seronegative pups were vaccinated at 8 weeks of age with modified-live canine distemper virus (CDV), canine adenovirus type-2 (CAV-2), and canine parvovirus (CPV) vaccine and at 12 weeks with a modified-live CDV, CAV-2, CPV, and killed rabies virus vaccine. An additional 31 seronegative pups served as age-matched, nonvaccinated controls. All test dogs were strictly isolated for 3 years after receiving the second vaccination and then were challenged with virulent rabies virus. Clinical signs of rabies were prevented in 28 (88%) of the 32 vaccinated dogs. In contrast, 97% (30 of 31) of the control dogs died of rabies infection. These study results indicated that no immunogenic interference occurred between the modified-live vaccine components and the killed rabies virus component. Furthermore, these results indicated that the rabies component in the test vaccine provided protection against virulent rabies challenge in dogs 12 weeks of age or older for a minimum of 3 years following vaccination.

  9. Cross-protection of a new type 2 porcine reproductive and respiratory syndrome virus (PRRSV) modified live vaccine (Fostera PRRS) against heterologous type 1 PRRSV challenge in growing pigs.

    PubMed

    Park, Changhoon; Choi, Kyuhyung; Jeong, Jiwoon; Chae, Chanhee

    2015-05-15

    The objective of the present study was to determine the cross-protection of a new type 2 porcine reproductive and respiratory syndrome virus (PRRSV) modified live vaccine against heterologous type 1 PRRSV challenge in growing pigs. The mean rectal temperature and respiratory score was significantly (P<0.05) lower in vaccinated challenged pigs than in unvaccinated challenged pigs. Vaccination of pigs with type 2 PRRSV reduced the levels of type 1 PRRSV viremia after challenge with type 1 PRRSV. Vaccinated challenged pigs had significantly (P<0.05) higher frequency of interferon-γ secreting cells and lower levels of interleukin-10 compared to unvaccinated challenged pigs. Vaccination of pigs with the type 2 PRRSV effectively reduced the macroscopic and microscopic lung lesion and the type 1 PRRSV antigens within lung lesions in vaccinated challenged pigs. This study demonstrates partial cross-protection of a new type 2 PRRSV modified live vaccine against heterologous type 1 PRRSV challenge in growing pigs. Copyright © 2015 Elsevier B.V. All rights reserved.

  10. 9 CFR 113.67 - Erysipelothrix Rhusiopathiae Vaccine.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Erysipelothrix Rhusiopathiae Vaccine... REQUIREMENTS Live Bacterial Vaccines § 113.67 Erysipelothrix Rhusiopathiae Vaccine. Erysipelothrix Rhusiopathiae Vaccine shall be prepared as a desiccated live culture of an avirulent or modified strain of...

  11. 9 CFR 113.67 - Erysipelothrix Rhusiopathiae Vaccine.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Erysipelothrix Rhusiopathiae Vaccine... REQUIREMENTS Live Bacterial Vaccines § 113.67 Erysipelothrix Rhusiopathiae Vaccine. Erysipelothrix Rhusiopathiae Vaccine shall be prepared as a desiccated live culture of an avirulent or modified strain of...

  12. 9 CFR 113.67 - Erysipelothrix Rhusiopathiae Vaccine.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Erysipelothrix Rhusiopathiae Vaccine... REQUIREMENTS Live Bacterial Vaccines § 113.67 Erysipelothrix Rhusiopathiae Vaccine. Erysipelothrix Rhusiopathiae Vaccine shall be prepared as a desiccated live culture of an avirulent or modified strain of...

  13. 9 CFR 113.67 - Erysipelothrix Rhusiopathiae Vaccine.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Erysipelothrix Rhusiopathiae Vaccine... REQUIREMENTS Live Bacterial Vaccines § 113.67 Erysipelothrix Rhusiopathiae Vaccine. Erysipelothrix Rhusiopathiae Vaccine shall be prepared as a desiccated live culture of an avirulent or modified strain of...

  14. 9 CFR 113.67 - Erysipelothrix Rhusiopathiae Vaccine.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Erysipelothrix Rhusiopathiae Vaccine... REQUIREMENTS Live Bacterial Vaccines § 113.67 Erysipelothrix Rhusiopathiae Vaccine. Erysipelothrix Rhusiopathiae Vaccine shall be prepared as a desiccated live culture of an avirulent or modified strain of...

  15. Modifying the 5'-Cap for Click Reactions of Eukaryotic mRNA and To Tune Translation Efficiency in Living Cells.

    PubMed

    Holstein, Josephin M; Anhäuser, Lea; Rentmeister, Andrea

    2016-08-26

    The 5'-cap is a hallmark of eukaryotic mRNAs and plays fundamental roles in RNA metabolism, ranging from quality control to export and translation. Modifying the 5'-cap may thus enable modulation of the underlying processes and investigation or tuning of several biological functions. A straightforward approach is presented for the efficient production of a range of N7-modified caps based on the highly promiscuous methyltransferase Ecm1. We show that these, as well as N(2) -modified 5'-caps, can be used to tune translation of the respective mRNAs both in vitro and in cells. Appropriate modifications allow subsequent bioorthogonal chemistry, as demonstrated by intracellular live-cell labeling of a target mRNA. The efficient and versatile N7 manipulation of the mRNA cap makes mRNAs amenable to both modulation of their biological function and intracellular labeling, and represents a valuable addition to the chemical biology toolbox. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Immunization of pups with maternally derived antibodies to canine parvovirus (CPV) using a modified-live variant (CPV-2b).

    PubMed

    Pratelli, A; Cavalli, A; Normanno, G; De Palma, M G; Pastorelli, G; Martella, V; Buonavoglia, C

    2000-05-01

    The results of vaccination trials carried out on pups with maternally derived antibodies (MDA) to canine parvovirus (CPV), using a modified-live CPV-2b variant vaccine (29-97/40 strain), are reported. The vaccine was able to overcome the obstacle of MDA, and to elicit protective immunity in 100% of the pups whose antibody titres were 1:10-1:40, 83% of the pups with titres of 1:80, 57% of the pups with titres of 1:160, and even in 60% of the pups with antibody titres of 1:320.

  17. Soy Intake Modifies the Relation Between Urinary Bisphenol A Concentrations and Pregnancy Outcomes Among Women Undergoing Assisted Reproduction.

    PubMed

    Chavarro, Jorge E; Mínguez-Alarcón, Lidia; Chiu, Yu-Han; Gaskins, Audrey J; Souter, Irene; Williams, Paige L; Calafat, Antonia M; Hauser, Russ

    2016-03-01

    Experimental data in rodents suggest that the adverse reproductive health effects of bisphenol A (BPA) can be modified by intake of soy phytoestrogens. Whether the same is true in humans is not known. The purpose of this study was to evaluate whether soy consumption modifies the relation between urinary BPA levels and infertility treatment outcomes among women undergoing assisted reproduction. The study was conducted in a fertility center in a teaching hospital. We evaluated 239 women enrolled between 2007 and 2012 in the Environment and Reproductive Health (EARTH) Study, a prospective cohort study, who underwent 347 in vitro fertilization (IVF) cycles. Participants completed a baseline questionnaire and provided up to 2 urine samples in each treatment cycle before oocyte retrieval. IVF outcomes were abstracted from electronic medical records. We used generalized linear mixed models with interaction terms to evaluate whether the association between urinary BPA concentrations and IVF outcomes was modified by soy intake. Live birth rates per initiated treatment cycle were measured. Soy food consumption modified the association of urinary BPA concentration with live birth rates (P for interaction = .01). Among women who did not consume soy foods, the adjusted live birth rates per initiated cycle in increasing quartiles of cycle-specific urinary BPA concentrations were 54%, 35%, 31%, and 17% (P for trend = .03). The corresponding live birth rates among women reporting pretreatment consumption of soy foods were 38%, 42%, 47%, and 49% (P for trend = 0.35). A similar pattern was found for implantation (P for interaction = .02) and clinical pregnancy rates (P for interaction = .03) per initiated cycle, where urinary BPA was inversely related to these outcomes among women not consuming soy foods but unrelated to them among soy consumers. Soy food intake may protect against the adverse reproductive effects of BPA. As these findings represent the first report suggesting a potential interaction between soy and BPA in humans, they should be further evaluated in other populations.

  18. Efficacy and safety of a modified-live cyprinid herpesvirus 3 vaccine in koi (Cyprinus carpio koi) for prevention of koi herpesvirus disease.

    PubMed

    Weber, E P Scott; Malm, Kirsten V; Yun, Susan C; Campbell, Lori A; Kass, Philip H; Marty, Gary D; Salonius, Kira; Dishon, Arnon

    2014-10-01

    To investigate safety and efficacy of a cyprinid herpesvirus type 3 (CyHV3) modified-live virus vaccine for the prevention of koi herpesvirus disease (KHVd). 420 healthy koi (Cyprinus carpio koi). Fish were vaccinated with a 1× dose or 10× overdose of CyHV3 modified-live virus vaccine or a placebo through bath exposure in tanks at 22°C. Horizontal transmission of vaccine virus was evaluated by commingling unvaccinated and vaccinated fish. Efficacy was evaluated by challenge exposure of vaccinated and naïve fish to a wild-type virus. Fish that died were submitted for quantitative PCR assay for CyHV3 and histologic evaluation. The CyHV3 vaccine was safe and efficacious, even at a 10× overdose. Vaccine-associated mortality rate was inversely associated with body weight, with a cumulative mortality rate of 9.4% (18/192) in fish weighing ≤ 87 g and no deaths in fish weighing > 87 g (0/48). Horizontal transfer of vaccine virus from vaccinates to naïve fish was negligible. For efficacy, the vaccine provided a significant reduction in mortality rate after challenge exposure to a wild-type virus, with a prevented fraction of 0.83 versus the placebo control fish. KHVd is highly contagious and commonly leads to deaths in 80% to 100% of exposed fish, representing a major threat to koi and common carp populations throughout the world. The CyHV3 modified-live virus vaccine had a favorable safety profile and was an effective vaccine for the control of KHVd in koi weighing > 87 g.

  19. Food insecurity, depression and the modifying role of social support among people living with HIV/AIDS in rural Uganda.

    PubMed

    Tsai, Alexander C; Bangsberg, David R; Frongillo, Edward A; Hunt, Peter W; Muzoora, Conrad; Martin, Jeffrey N; Weiser, Sheri D

    2012-06-01

    Depression is common among people living with HIV/AIDS and contributes to a wide range of worsened HIV-related outcomes, including AIDS-related mortality. Targeting modifiable causes of depression, either through primary or secondary prevention, may reduce suffering as well as improve HIV-related outcomes. Food insecurity is a pervasive source of uncertainty for those living in resource-limited settings, and cross-sectional studies have increasingly recognized it as a critical determinant of poor mental health. Using cohort data from 456 men and women living with HIV/AIDS initiating HIV antiretroviral therapy in rural Uganda, we sought to (a) estimate the association between food insecurity and depression symptom severity, (b) assess the extent to which social support may serve as a buffer against the adverse effects of food insecurity, and (c) determine whether the buffering effects are specific to certain types of social support. Quarterly data were collected by structured interviews and blood draws. The primary outcome was depression symptom severity, measured by a modified Hopkins Symptom Checklist for Depression. The primary explanatory variables were food insecurity, measured with the Household Food Insecurity Access Scale, and social support, measured with a modified version of the Functional Social Support Questionnaire. We found that food insecurity was associated with depression symptom severity among women but not men, and that social support buffered the impacts of food insecurity on depression. We also found that instrumental support had a greater buffering influence than emotional social support. Interventions aimed at improving food security and strengthening instrumental social support may have synergistic beneficial effects on both mental health and HIV outcomes among PLWHA in resource-limited settings. Copyright © 2012 Elsevier Ltd. All rights reserved.

  20. Effects of North American Porcine Reproductive and Respiratory Syndrome Virus (PRRSV)-Based Modified Live Vaccines on Preimmunized Sows Artificially Inseminated with European PRRSV-Spiked Semen

    PubMed Central

    Han, Kiwon; Seo, Hwi Won; Oh, Yeonsu; Kang, Ikjae; Park, Changhoon

    2012-01-01

    The objective of the present study was to determine if the European porcine reproductive and respiratory syndrome virus (PRRSV) can be transmitted via spiked semen to preimmunized sows and induce reproductive failure. Sows were immunized with the North American PRRSV-based modified live vaccine (Ingelvac PRRS MLV; Boehringer Ingelheim Animal Health, St. Joseph, MO) and were artificially inseminated. The sows were randomly divided into three groups. The vaccinated (group 2) and nonvaccinated (group 3) sows developed a PRRSV viremia at 7 to 28 days postinsemination with the European PRRSV-spiked semen. The number of genomic copies of the European PRRSV in serum samples was not significantly different between vaccinated and nonvaccinated sows. All negative-control sows in group 1 farrowed at the expected date. The sows in groups 2 and 3 farrowed between 103 and 110 days after the first insemination. European PRRSV RNA was detected in the lungs of 8 out of 11 live-born piglets and 46 out of 54 stillborn fetuses. In addition, PRRSV RNA was detected using in situ hybridization in other tissues from vaccinated sows that had been inseminated with European PRRSV-spiked semen (group 2). The present study has demonstrated that vaccinating sows with the North American PRRSV-based modified live vaccine does not prevent reproductive failure after insemination with European PRRSV-spiked semen. PMID:22237898

  1. Effects of North American porcine reproductive and respiratory syndrome virus (PRRSV)-based modified live vaccines on preimmunized sows artificially inseminated with European PRRSV-spiked semen.

    PubMed

    Han, Kiwon; Seo, Hwi Won; Oh, Yeonsu; Kang, Ikjae; Park, Changhoon; Chae, Chanhee

    2012-03-01

    The objective of the present study was to determine if the European porcine reproductive and respiratory syndrome virus (PRRSV) can be transmitted via spiked semen to preimmunized sows and induce reproductive failure. Sows were immunized with the North American PRRSV-based modified live vaccine (Ingelvac PRRS MLV; Boehringer Ingelheim Animal Health, St. Joseph, MO) and were artificially inseminated. The sows were randomly divided into three groups. The vaccinated (group 2) and nonvaccinated (group 3) sows developed a PRRSV viremia at 7 to 28 days postinsemination with the European PRRSV-spiked semen. The number of genomic copies of the European PRRSV in serum samples was not significantly different between vaccinated and nonvaccinated sows. All negative-control sows in group 1 farrowed at the expected date. The sows in groups 2 and 3 farrowed between 103 and 110 days after the first insemination. European PRRSV RNA was detected in the lungs of 8 out of 11 live-born piglets and 46 out of 54 stillborn fetuses. In addition, PRRSV RNA was detected using in situ hybridization in other tissues from vaccinated sows that had been inseminated with European PRRSV-spiked semen (group 2). The present study has demonstrated that vaccinating sows with the North American PRRSV-based modified live vaccine does not prevent reproductive failure after insemination with European PRRSV-spiked semen.

  2. Effective intracellular inhibition of the cAMP-dependent protein kinase by microinjection of a modified form of the specific inhibitor peptide PKi in living fibroblasts.

    PubMed

    Fernandez, A; Mery, J; Vandromme, M; Basset, M; Cavadore, J C; Lamb, N J

    1991-08-01

    In order to obtain a peptide retaining its biological activity following microinjection into living cells, we have modified a synthetic peptide [PKi(m)(6-24)], derived from the specific inhibitor protein of the cAMP-dependent protein kinase (A-kinase) in two ways: (1) substitution of the arginine at position 18 for a D-arginine; (2) blockade of the side chain on the C-terminal aspartic acid by a cyclohexyl ester group. In an in vitro assay, PKi(m) has retained a specific inhibitory activity against A-kinase as assessed against six other kinases, with similar efficiency to that of the unmodified PKi(5-24) peptide. Microinjection of PKi(m) into living fibroblasts reveals its capacity to prevent the changes in cell morphology and cytoskeleton induced by drugs which activate endogenous A-kinase, whereas the original PKi peptide failed to do so. This inhibition of A-kinase in vivo by PKi(m) lasts between 4 and 6 h after injection. In light of its effective half-life, this modified peptide opens a route for the use of biologically active peptides in vivo, an approach which has been hampered until now by the exceedingly short half-life of peptides inside living cells. By providing a direct means of inhibiting A-kinase activity for sufficiently long periods to observe effects on cellular functions in living cells, PKi(m) represents a powerful tool in studying the potential role of cAMP-dependent phosphorylation in vivo.

  3. Foreign-Born Concentration and Acculturation to Volunteering among Immigrant Youth

    ERIC Educational Resources Information Center

    Tong, Yuying

    2010-01-01

    Using children of immigrants sample from National Longitudinal Study of Adolescent Health, this study investigates how immigrant youth acculturating to the American social norm of volunteering and how the acculturation is modified by living in immigrant neighborhoods. Multilevel logistic regression produces distinct patterns for children living in…

  4. Attempt to develop live attenuated bacterial vaccines by selecting resistance to gossypol, proflavine hemisulfate, novobiocin, or ciprofloxacin

    USDA-ARS?s Scientific Manuscript database

    In an attempt to develop attenuated bacteria as potential live vaccines, four chemicals (gossypol, proflavine hemisulfate, novobiocin, and ciprofloxacin) were used to modify the following four genera of bacteria through chemical-resistance strategy: 1) Aeromonas hydrophila (9 isolates); 2) Edwardsie...

  5. Attempt to develop live attenuated bacterial vaccines by selecting resistance to gossypol, proflavine hemisulfate, novobiocin, or ciprofloxacin

    USDA-ARS?s Scientific Manuscript database

    In an attempt to develop attenuated bacteria as potential live vaccines, four chemicals (gossypol, proflavine hemisulfate, novobiocin, and ciprofloxacin) were used to modify the following four genera of bacteria through chemical-resistance strategy: (1) Aeromonas hydrophila (9 isolates); (2) Edwards...

  6. 9 CFR 113.70 - Pasteurella Multocida Vaccine, Avian Isolate.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Pasteurella Multocida Vaccine, Avian... REQUIREMENTS Live Bacterial Vaccines § 113.70 Pasteurella Multocida Vaccine, Avian Isolate. Pasteurella Multocida Vaccine, Avian Isolate, shall be prepared as a desiccated live culture of an avirulent or modified...

  7. 9 CFR 113.70 - Pasteurella Multocida Vaccine, Avian Isolate.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Pasteurella Multocida Vaccine, Avian... REQUIREMENTS Live Bacterial Vaccines § 113.70 Pasteurella Multocida Vaccine, Avian Isolate. Pasteurella Multocida Vaccine, Avian Isolate, shall be prepared as a desiccated live culture of an avirulent or modified...

  8. 9 CFR 113.70 - Pasteurella Multocida Vaccine, Avian Isolate.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Pasteurella Multocida Vaccine, Avian... REQUIREMENTS Live Bacterial Vaccines § 113.70 Pasteurella Multocida Vaccine, Avian Isolate. Pasteurella Multocida Vaccine, Avian Isolate, shall be prepared as a desiccated live culture of an avirulent or modified...

  9. 9 CFR 113.70 - Pasteurella Multocida Vaccine, Avian Isolate.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Pasteurella Multocida Vaccine, Avian... REQUIREMENTS Live Bacterial Vaccines § 113.70 Pasteurella Multocida Vaccine, Avian Isolate. Pasteurella Multocida Vaccine, Avian Isolate, shall be prepared as a desiccated live culture of an avirulent or modified...

  10. 9 CFR 113.70 - Pasteurella Multocida Vaccine, Avian Isolate.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Pasteurella Multocida Vaccine, Avian... REQUIREMENTS Live Bacterial Vaccines § 113.70 Pasteurella Multocida Vaccine, Avian Isolate. Pasteurella Multocida Vaccine, Avian Isolate, shall be prepared as a desiccated live culture of an avirulent or modified...

  11. Variation in levels of anxiety to dental treatment among nonorphan and orphan children living under different systems.

    PubMed

    Chikkala, Jayanth; Chandrabhatla, Srinivas Kumar; Vanga, Narasimha Rao V

    2015-08-01

    It is essential to understand the factors influencing the level of anxiety to dental treatment among different children as it can influence seeking dental care. Here, we assessed the impact of parental loss on dental anxiety among 6-13-year-old children. A total of 444 children within the age group 6-13 years were selected. Group 1 consisted of orphan children living in government-run orphanages, Group 2 consisted of orphan children taken care by a person with a motherly relationship, Group 3 consisted of abandoned children living in private organization and Group 4 consisted of children living with their parents. Dental anxiety was measured using children's fear survey schedule-dental subscale and modified faces version of modified child dental anxiety scale. The highest number of anxious children were observed in Group 4 and the difference in the anxiety levels among the four groups was found to be highly statistically significant. Children living in government-run orphanages had least dental anxiety. All the orphans may not have the same anxiety levels and the environment of upbringing the orphans plays a significant role in the development of the anxiety.

  12. In Cell Footprinting Coupled with Mass Spectrometry for the Structural Analysis of Proteins in Live Cells.

    PubMed

    Espino, Jessica A; Mali, Vishaal S; Jones, Lisa M

    2015-08-04

    Protein footprinting coupled with mass spectrometry has become a widely used tool for the study of protein-protein and protein-ligand interactions and protein conformational change. These methods provide residue-level analysis on protein interaction sites and have been successful in studying proteins in vitro. The extension of these methods for in cell footprinting would open an avenue to study proteins that are not amenable for in vitro studies and would probe proteins in their native environment. Here we describe the application of an oxidative-based footprinting approach inside cells in which hydroxyl radicals are used to oxidatively modify proteins. Mass spectrometry is used to detect modification sites and to calculate modification levels. The method is probing biologically relevant proteins in live cells, and proteins in various cellular compartments can be oxdiatively modified. Several different amino acid residues are modified making the method a general labeling strategy for the study of a variety of proteins. Further, comparison of the extent of oxidative modification with solvent accessible surface area reveals the method successfully probes solvent accessibility. This marks the first time protein footprinting has been performed in live cells.

  13. Use of genetically modified bacteria for drug delivery in humans: Revisiting the safety aspect.

    PubMed

    Wegmann, Udo; Carvalho, Ana Lucia; Stocks, Martin; Carding, Simon R

    2017-05-23

    The use of live, genetically modified bacteria as delivery vehicles for biologics is of considerable interest scientifically and has attracted significant commercial investment. We have pioneered the use of the commensal gut bacterium Bacteroides ovatus for the oral delivery of therapeutics to the gastrointestinal tract. Here we report on our investigations of the biological safety of engineered B. ovatus bacteria that includes the use of thymineless death as a containment strategy and the potential for the spread of transgenes in vivo in the mammalian gastrointestinal tract. We demonstrate the ability of GM-strains of Bacteroides to survive thymine starvation and overcome it through the exchange of genetic material. We also provide evidence for horizontal gene transfer in the mammalian gastrointestinal tract resulting in transgene-carrying wild type bacteria. These findings sound a strong note of caution on the employment of live genetically modified bacteria for the delivery of biologics.

  14. Does the use of gonadotropin-releasing hormone antagonists in natural IVF cycles for poor responder patients cause more harm than benefit?

    PubMed

    Aksoy, Senai; Yakin, Kayhan; Seyhan, Ayse; Oktem, Ozgur; Alatas, Cengiz; Ata, Baris; Urman, Bulent

    2016-06-01

    Poor ovarian response to controlled ovarian stimulation (COS) is one of the most critical factors that substantially limits the success of assisted reproduction techniques (ARTs). Natural and modified natural cycle IVF are two options that could be considered as a last resort. Blocking gonadotropin-releasing hormone (GnRH) actions in the endometrium via GnRH receptor antagonism may have a negative impact on endometrial receptivity. We analysed IVF outcomes in 142 natural (n = 30) or modified natural (n = 112) IVF cycles performed in 82 women retrospectively. A significantly lower proportion of natural cycles reached follicular aspiration compared to modified natural cycles (56.7% vs. 85.7%, p < 0.001). However, the difference between the numbers of IVF cycles ending in embryo transfer (26.7% vs. 44.6%) was not statistically significant between natural cycle and modified natural IVF cycles. Clinical pregnancy (6.7% vs. 7.1%) and live birth rates per initiated cycle (6.7% vs. 5.4%) were similar between the two groups. Notably, the implantation rate was slightly lower in modified natural cycles (16% vs. 25%, p > 0.05). There was a trend towards higher clinical pregnancy (25% vs. 16%) and live birth (25% vs. 12%) rates per embryo transfer in natural cycles compared to modified natural cycles, but the differences did not reach statistical significance.

  15. Development of live attenuated sparfloxacin-resistant Streptococcus agalactiae polyvalent vaccines to protect Nile tilapia

    USDA-ARS?s Scientific Manuscript database

    To develop attenuated bacteria as potential live vaccines, sparfloxacin was used in this study to modify 40 isolates of Streptococcus agalactiae. Majority of S. agalactiae used in this study were able to develop at least 80-fold resistance to sparfloxacin. When the virulence of the sparfloxacin-resi...

  16. Development of live attenuated Streptococcus agalactiae as potential vaccines by selecting for resistance to sparfloxacin

    USDA-ARS?s Scientific Manuscript database

    To develop attenuated bacteria as potential live vaccines, sparfloxacin was used in this study to modify 40 isolates of Streptococcus agalactiae. Majority of S. agalactiae used in this study were able to develop at least 80-fold resistance to sparfloxacin. When the virulence of the sparfloxacin-resi...

  17. Fatal vaccine-induced canine distemper virus infection in black-footed ferrets

    USGS Publications Warehouse

    Carpenter, J.W.; Appel, M.J.G.; Erickson, R.C.; Novilla, M.N.

    1976-01-01

    Four black-footed ferrets that were live-trapped in South Dakota and transported to the Patuxent Wildlife Research Center died within 21 days after vaccination with modified live canine distemper virus. Immunofluorescence, European ferret inoculation, virus isolation attempts, and serum-neutralization tests indicated insufficient attenuation of the vaccine for this species.

  18. Fatal vaccine-induced canine distemper virus infection in black-footed ferrets.

    PubMed

    Carpenter, J W; Appel, M J; Erickson, R C; Novilla, M N

    1976-11-01

    Four black-footed ferrets that were live-trapped in South Dakota and transported to the Patuxent Wildlife Research Center died within 21 days after vaccination with modified live canine distemper virus. Immunofluorescence, European ferret inoculation, virus isolation attempts, and serum-neutralization tests indicated insufficient attenuation of the vaccine for this species.

  19. Miniature fiber optic spectrometer-based quantitative fluorescence resonance energy transfer measurement in single living cells.

    PubMed

    Chai, Liuying; Zhang, Jianwei; Zhang, Lili; Chen, Tongsheng

    2015-03-01

    Spectral measurement of fluorescence resonance energy transfer (FRET), spFRET, is a widely used FRET quantification method in living cells today. We set up a spectrometer-microscope platform that consists of a miniature fiber optic spectrometer and a widefield fluorescence microscope for the spectral measurement of absolute FRET efficiency (E) and acceptor-to-donor concentration ratio (R(C)) in single living cells. The microscope was used for guiding cells and the spectra were simultaneously detected by the miniature fiber optic spectrometer. Moreover, our platform has independent excitation and emission controllers, so different excitations can share the same emission channel. In addition, we developed a modified spectral FRET quantification method (mlux-FRET) for the multiple donors and multiple acceptors FRET construct (mD∼nA) sample, and we also developed a spectra-based 2-channel acceptor-sensitized FRET quantification method (spE-FRET). We implemented these modified FRET quantification methods on our platform to measure the absolute E and R(C) values of tandem constructs with different acceptor/donor stoichiometries in single living Huh-7 cells.

  20. A recombinase polymerase amplification-based assay for rapid detection of African swine fever virus.

    PubMed

    Wang, Jianchang; Wang, Jinfeng; Geng, Yunyun; Yuan, Wanzhe

    2017-10-01

    A recombinase polymerase amplification (RPA)-based method was developed for rapid and specific detection of African swine fever virus (ASFV), the etiologic agent of African swine fever, a devastating disease of swine. Primers and the exo probe targeting the conserved region of the P72 gene of ASFV were designed and the reaction was run on the Genie III scanner device. Using recombinant plasmid DNA containing the P72 gene as template, we showed that the amplified product could be detected in less than 10 min and that the detection limit was 10 2 copies DNA/reaction [same detection limit as real-time polymerase chain reaction (PCR)]. The RPA assay did not cross-detect CSFV, PCV-2, PRV, PRRSV, or FMDV, common viruses seen in pigs. Tests of recombinant plasmid-spiked serum samples revealed that RPA and real-time PCR had the same diagnostic rate. The RPA assay, which is simple, cost-effective, and fast, is a promising alternative to real-time PCR for ASFV detection.

  1. A recombinase polymerase amplification-based assay for rapid detection of African swine fever virus

    PubMed Central

    Wang, Jianchang; Wang, Jinfeng; Geng, Yunyun; Yuan, Wanzhe

    2017-01-01

    A recombinase polymerase amplification (RPA)-based method was developed for rapid and specific detection of African swine fever virus (ASFV), the etiologic agent of African swine fever, a devastating disease of swine. Primers and the exo probe targeting the conserved region of the P72 gene of ASFV were designed and the reaction was run on the Genie III scanner device. Using recombinant plasmid DNA containing the P72 gene as template, we showed that the amplified product could be detected in less than 10 min and that the detection limit was 102 copies DNA/reaction [same detection limit as real-time polymerase chain reaction (PCR)]. The RPA assay did not cross-detect CSFV, PCV-2, PRV, PRRSV, or FMDV, common viruses seen in pigs. Tests of recombinant plasmid-spiked serum samples revealed that RPA and real-time PCR had the same diagnostic rate. The RPA assay, which is simple, cost-effective, and fast, is a promising alternative to real-time PCR for ASFV detection. PMID:29081590

  2. Soy Intake Modifies the Relation Between Urinary Bisphenol A Concentrations and Pregnancy Outcomes Among Women Undergoing Assisted Reproduction

    PubMed Central

    Mínguez-Alarcón, Lidia; Chiu, Yu-Han; Gaskins, Audrey J.; Souter, Irene; Williams, Paige L.; Calafat, Antonia M.; Hauser, Russ

    2016-01-01

    Context: Experimental data in rodents suggest that the adverse reproductive health effects of bisphenol A (BPA) can be modified by intake of soy phytoestrogens. Whether the same is true in humans is not known. Objective: The purpose of this study was to evaluate whether soy consumption modifies the relation between urinary BPA levels and infertility treatment outcomes among women undergoing assisted reproduction. Setting: The study was conducted in a fertility center in a teaching hospital. Design: We evaluated 239 women enrolled between 2007 and 2012 in the Environment and Reproductive Health (EARTH) Study, a prospective cohort study, who underwent 347 in vitro fertilization (IVF) cycles. Participants completed a baseline questionnaire and provided up to 2 urine samples in each treatment cycle before oocyte retrieval. IVF outcomes were abstracted from electronic medical records. We used generalized linear mixed models with interaction terms to evaluate whether the association between urinary BPA concentrations and IVF outcomes was modified by soy intake. Main Outcome Measure: Live birth rates per initiated treatment cycle were measured. Results: Soy food consumption modified the association of urinary BPA concentration with live birth rates (P for interaction = .01). Among women who did not consume soy foods, the adjusted live birth rates per initiated cycle in increasing quartiles of cycle-specific urinary BPA concentrations were 54%, 35%, 31%, and 17% (P for trend = .03). The corresponding live birth rates among women reporting pretreatment consumption of soy foods were 38%, 42%, 47%, and 49% (P for trend = 0.35). A similar pattern was found for implantation (P for interaction = .02) and clinical pregnancy rates (P for interaction = .03) per initiated cycle, where urinary BPA was inversely related to these outcomes among women not consuming soy foods but unrelated to them among soy consumers. Conclusion: Soy food intake may protect against the adverse reproductive effects of BPA. As these findings represent the first report suggesting a potential interaction between soy and BPA in humans, they should be further evaluated in other populations. PMID:26815879

  3. Repeated suppression of lymphocyte blastogenesis following vaccinations of CPV-immune dogs with modified-live CPV vaccines.

    PubMed

    Mastro, J M; Axthelm, M; Mathes, L E; Krakowka, S; Ladiges, W; Olsen, R G

    1986-09-01

    A commercially available modified-live canine parvovirus (CPV) vaccine was evaluated for its immunosuppressive properties in eight random-bred dogs, all with circulatory antibody to CPV. Three of the eight dogs exhibited a significant decrease in lymphocyte blastogenesis after vaccine administration. In these dogs, this decrease in blastogenesis was of short duration and was consistently observed after repeated administrations of the vaccine. Neither gastroenteritis, fever nor leukopenia, signs indicative of virulent canine parvovirus infection, were detected in these animals. In addition, lymphocytes from these dogs lacked Ia antigen expression. This study demonstrated that the immunomodulating effects of ML-CPV is not observed in all animals yet is consistent in affected individuals.

  4. Nuclear isospin effect on α-decay half-lives

    NASA Astrophysics Data System (ADS)

    Akrawy, Dashty T.; Hassanabadi, H.; Hosseini, S. S.; Santhosh, K. P.

    2018-07-01

    The α-decay half-lives for the even-even, even-odd, odd-even and odd-odd of 356 nuclei in the range 52 ≤Zp ≤ 118 have been studied within the analytical formula of Royer and also within the modified analytical formula of Royer. We calculated the new coefficient of the Royer by fitting 356 isotopes. Also, we considered the Denisov and Khudenko formula and obtained the new coefficient for the modified Denisov and Khudenko formula. We calculated the standard deviation and the average deviation. Analytical results are compared with the experimental data. The results are in better agreement with the experimental data when the effect of the isospin considered for the parent nuclei.

  5. In vitro proteolysis of nematode FLPs by preparations from the free-living nematode Panagrellus redivivus and two plant-parasitic nematodes (Heterodera glycines and Meloidogyne incognita)

    USDA-ARS?s Scientific Manuscript database

    Proteolytic activities in extracts from three nematodes, the plant parasites Heterodera glycines and Meloidogyne incognita, and the free-living Panagrellus redivivus, were surveyed for substrate preferences using a battery of seven FRET-modified peptide substrates, all derived from members of the la...

  6. Synthesis and Evaluation of Novel RSK Inhibitors in a Living Human Breast Model

    DTIC Science & Technology

    2015-09-01

    the n- propyl carbamate analogue 9, and the sec-butyl carbamate analogue 10. The modified synthetic route could also be applied to the synthesis of...AWARD NUMBER: W81XWH-11-1-0068 TITLE: Synthesis and Evaluation of Novel RSK Inhibitors in a Living Human Breast Model PRINCIPAL INVESTIGATOR...DATES COVERED 01 Jul 2011 – 30 Jun 2015 4. TITLE AND SUBTITLE Synthesis and Evaluation of Novel RSK Inhibitors in a Living Human Breast Model 5a

  7. Bovine viral diarrhea virus fetal persistent infection after immunization with a contaminated modified-live virus vaccine.

    PubMed

    Palomares, Roberto A; Marley, Shonda M; Givens, M Daniel; Gallardo, Rodrigo A; Brock, Kenny V

    2013-05-01

    The objective was to determine whether a multivalent modified-live virus vaccine containing noncytopathic bovine viral diarrhea virus (BVDV) administered off-label to pregnant cattle can result in persistently infected fetuses and to assess whether vaccinal strains can be shed to unvaccinated pregnant cattle commingling with vaccinates. Nineteen BVDV-naïve pregnant heifers were randomly assigned to two groups: cattle vaccinated near Day 77 of gestation with modified-live virus vaccine containing BVDV-1a (WRL strain), bovine herpes virus-1, parainfluenza 3, and bovine respiratory syncytial virus (Vx group; N = 10) or control unvaccinated cattle (N = 9). During the course of the study a voluntary stop-sale/recall was conducted by the manufacturer because of the presence of a BVDV contaminant in the vaccine. At Day 175 of gestation, fetuses were removed by Cesarean section and fetal tissues were submitted for virus isolation, and quantitative reverse transcription polymerase chain reaction using BVDV-1- and BVDV-2-specific probes. Nucleotide sequencing of viral RNA was performed for quantitative reverse transcription polymerase chain reaction-positive samples. Two vaccinated and two control heifers aborted their pregnancies, but their fetuses were unavailable for BVDV testing. Virus was isolated from all eight fetuses in the Vx group heifers and from 2 of 7 fetuses in the control unvaccinated heifers. Only BVDV-2 was detected in fetuses from the Vx group, and only BVDV-1 was detected in the two fetuses from the control group. Both BVDV-1 and BVDV-2 were detected in the vaccine. In conclusion, vaccination of pregnant heifers with a contaminated modified-live BVDV vaccine resulted in development of BVDV-2 persistently infected fetuses in all tested vaccinated animals. Furthermore, BVDV was apparently shed to unvaccinated heifers causing fetal infections from which only BVDV-1 was detected. Published by Elsevier Inc.

  8. One year duration of immunity of the modified live bovine viral diarrhea virus type 1 and type 2 and bovine herpesvirus-1 fractions of Vista® Once SQ vaccine.

    PubMed

    Purtle, Lisa; Mattick, Debra; Schneider, Corey; Smith, Linda; Xue, Wenzhi; Trigo, Emilio

    2016-03-18

    Three studies were performed to determine the duration of immunity of the bovine viral diarrhea virus type 1 and type 2 (BVDV-1 and BVDV-2) and bovine herpesvirus-1 (BHV-1) fractions of a commercially prepared modified-live vaccine. Vista® Once SQ (Vista®) vaccine contains five modified-live viruses, BVDV-1, BVDV-2, BHV-1, bovine respiratory syncytial virus, and bovine parainfluenza 3 virus, and two modified-live bacteria, Pasteurella multocida and Mannheimia haemolytica. For all three studies, calves were administered a single dose of vaccine or placebo vaccine subcutaneously, and were challenged with one of the three virulent viruses at least one year following vaccination. Calves were evaluated daily following challenge for clinical signs of disease associated with viral infection, nasal swab samples were evaluated for virus shedding, and serum was tested for neutralizing antibodies. Following the BVDV-1 and BVDV-2 challenges, whole blood was evaluated for white blood cell counts, and for the BVDV-2 study, whole blood was also evaluated for platelet counts. Calves vaccinated with BVDV type 1a, were protected from challenge with BVDV type 1b, and had significant reductions in clinical disease, fever, leukopenia, and virus shedding compared to control calves. Vaccinated calves in the BVDV-2 study were protected from clinical disease, mortality, fever, leukopenia, thrombocytopenia, and virus shedding compared to controls. Vaccinated calves in the BHV-1 study were protected from clinical disease and fever, and had significantly reduced duration of nasal virus shedding. These three studies demonstrated that a single administration of the Vista® vaccine to healthy calves induces protective immunity against BVDV-1, BVDV-2 and BHV-1 that lasts at least one year following vaccination. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

  9. Radical SAM Enzymes in the Biosynthesis of Ribosomally Synthesized and Post-translationally Modified Peptides (RiPPs).

    PubMed

    Benjdia, Alhosna; Balty, Clémence; Berteau, Olivier

    2017-01-01

    Ribosomally-synthesized and post-translationally modified peptides (RiPPs) are a large and diverse family of natural products. They possess interesting biological properties such as antibiotic or anticancer activities, making them attractive for therapeutic applications. In contrast to polyketides and non-ribosomal peptides, RiPPs derive from ribosomal peptides and are post-translationally modified by diverse enzyme families. Among them, the emerging superfamily of radical SAM enzymes has been shown to play a major role. These enzymes catalyze the formation of a wide range of post-translational modifications some of them having no counterparts in living systems or synthetic chemistry. The investigation of radical SAM enzymes has not only illuminated unprecedented strategies used by living systems to tailor peptides into complex natural products but has also allowed to uncover novel RiPP families. In this review, we summarize the current knowledge on radical SAM enzymes catalyzing RiPP post-translational modifications and discuss their mechanisms and growing importance notably in the context of the human microbiota.

  10. A Review of Living Collections with Special Emphasis on Sustainability and Its Impact on Research Across Multiple Disciplines

    PubMed Central

    2017-01-01

    Formal living collections have unique characteristics that distinguish them from other types of biorepositories. Comprising diverse resources, microbe culture collections, crop and biodiversity plant germplasm collections, and animal germplasm repositories are commonly allied with specific research communities or stakeholder groups. Among living collections, microbial culture collections have very long and unique life histories, with some being older than 100 years. Regulatory, financial, and technical developments have impacted living collections in many ways. International treaty obligations and restrictions on release of genetically modified organisms complicate the activities of living collections. Funding for living collections is a continuing challenge and threatens to create a two-tier system where medically relevant collections are well funded and all other collections are underfunded and hence understaffed. Molecular, genetic, and whole genome sequence analysis of contents of microbes and other living resource collections bring additional value to living collections. PMID:27869477

  11. A Review of Living Collections with Special Emphasis on Sustainability and Its Impact on Research Across Multiple Disciplines.

    PubMed

    McCluskey, Kevin

    2017-02-01

    Formal living collections have unique characteristics that distinguish them from other types of biorepositories. Comprising diverse resources, microbe culture collections, crop and biodiversity plant germplasm collections, and animal germplasm repositories are commonly allied with specific research communities or stakeholder groups. Among living collections, microbial culture collections have very long and unique life histories, with some being older than 100 years. Regulatory, financial, and technical developments have impacted living collections in many ways. International treaty obligations and restrictions on release of genetically modified organisms complicate the activities of living collections. Funding for living collections is a continuing challenge and threatens to create a two-tier system where medically relevant collections are well funded and all other collections are underfunded and hence understaffed. Molecular, genetic, and whole genome sequence analysis of contents of microbes and other living resource collections bring additional value to living collections.

  12. Live long and prosper? Childhood living conditions, marital status, social class in adulthood and mortality during mid-life: a cohort study.

    PubMed

    Fors, Stefan; Lennartsson, Carin; Lundberg, Olle

    2011-03-01

    The aim of the present study was to investigate the impact of childhood living conditions, marital status, and social class in adulthood on the risk of mortality during mid-life. Two questions were addressed: Is there an effect of childhood living conditions on mortality risk during mid-life and if so, is the effect mediated or modified by social class and/or marital status in adulthood? A nationally representative, Swedish, level of living survey from 1968 was used as baseline. The study included those aged 25-69 at baseline (n = 4082). Social conditions in childhood and adulthood were assessed using self-reports. These individuals were then followed for 39 years using registry data on mortality. The results showed associations between childhood living conditions, marital status, social class in adulthood and mortality during mid life. Social class and familial conditions during childhood as well as marital status and social class in adulthood all contributed to the risk of mortality during mid-life. Individuals whose father's were manual workers, who grew up in broken homes, who were unmarried, and/or were manual workers in adulthood had an increased risk of mortality during mid life. The effects of childhood conditions were, in part, both mediated and modified by social class in adulthood. The findings of this study suggest that there are structural, social conditions experienced at different stages of the life course that affect the risk of mortality during mid-life.

  13. Employees with Cerebral Palsy

    MedlinePlus

    ... Allowing use of a service animal Allowing extra time for activities of daily living (ADL) Fine Motor Impairment: Modifying workstation design Using alternative computer input devices/software Using telephone ...

  14. Development of a highly sensitive PCR/DNA chip method to detect mycoplasmas in a veterinary modified live vaccine.

    PubMed

    Mbelo, Sylvie; Gay, Virginie; Blanchard, Stephanie; Abachin, Eric; Falque, Stephanie; Lechenet, Jacques; Poulet, Hervé; de Saint-Vis, Blandine

    2018-05-09

    Mycoplasmas are potential contaminants that introduce undesirable changes in mammalian cell cultures. They frequently contaminate cell substrates and other starting materials used for manufacturing cell-derived biologics, such as vaccines and pharmaceutical products. Mycoplasma purity testing of live vaccines, active ingredients, raw material, and seed lots is required during vaccine production. Previously, testing using a time-consuming, costly 28-day culture assay, which lacks sensitivity for species that do not grow in culture, was required in the European Pharmacopoeia (Ph. Eur). But now nucleic acid amplification techniques (NATs) can be used. NATs provide rapid results and are sensitive. We evaluated the sensitivity and specificity of a commercially-available NAT to detect individual mycoplasma DNA in a veterinary modified live vaccine using five reference strains recommended by the Ph. Eur. Our results showed that this NAT-based method can be used to detect mycoplasma in spiked live vaccine, without interference from the vaccine components, with a limit of detection of 10 CFU/mL, as required by the Ph. Eur. Its specificity was demonstrated since no mycoplasmas were detected in non-spiked vaccine. This method is undergoing validation as a replacement for the conventional culture method in the production of veterinary live vaccines. Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.

  15. Complex outflow anatomy in left lateral lobe graft and modified venous reconstruction in pediatric living donor liver transplantation.

    PubMed

    Benetatos, Nikolaos; Scalera, Irene; Isaac, John R; Mirza, Darius F; Muiesan, Paolo

    2014-10-01

    Hepatic venous outflow reconstruction is of critical significance in pediatric patients undergoing living donor liver transplantation. Accurate knowledge of the anatomical variations is important to obtain appropriate size segmental grafts. The diameter of the hepatic veins and the potential risk of complications at the level of the anastomosis require an adequate primary vascular reconstruction. We describe a venous outflow reconstruction technique, in a living related left lateral lobe graft, with unfavorable hepatic venous anatomy. © 2014 Steunstichting ESOT.

  16. "Maybe the Algae Was from the Filter": Maybe and Similar Modifiers as Mediational Tools and Indicators of Uncertainty and Possibility in Children's Science Talk

    ERIC Educational Resources Information Center

    Kirch, Susan A.; Siry, Christina A.

    2012-01-01

    Uncertainty is an essential component of scientific inquiry and it also permeates our daily lives. Understanding how to identify, evaluate, resolve and live in the presence of uncertainty is important for decision-making strategies and engaging in transformative actions. In contrast, confidence and certainty are prized in elementary school…

  17. Avoiding genetically modified foods in GMO Ground Zero: A reflective self-narrative.

    PubMed

    Edwards, Sachi

    2015-05-01

    I engage in a reflective self-narrative of my experience attempting to maintain a diet free of genetically modified organisms. Social tension over the genetically modified organism industry in Hawai'i, United States, has led to public debates over jobs and social identities. Drawing on local media sources, grassroots organizations, and blog posts, I describe the way this tension has shaped my experience with food, eating, and being with others as a genetically modified organism avoider. I utilize discursive positioning to make sense of my experiences by locating them within the ongoing public conversations that give structure to the daily lives of Hawai'i's residents. © The Author(s) 2015.

  18. Preliminary development of a live attenuated canine parvovirus vaccine from an isolate of British origin.

    PubMed

    Churchill, A E

    1987-04-04

    Canine parvovirus isolated from a case of haemorrhagic enteritis in a breeding kennel in England was passaged and cloned in cultured feline and canine cells. No significant evidence of pathogenicity was found during six serial passages of the modified virus back through young dogs. The attenuated virus was excreted by inoculated animals and spread rapidly to uninoculated animals held in contact. When high titre attenuated virus was given to the six-week-old offspring of a seropositive dam a prompt seroconversion was observed. When the attenuated virus was used as an experimental vaccine in 108 pups in an infected breeding colony a highly significant improvement was obtained in the accumulated morbidity and mortality compared with a parallel group vaccinated with modified live feline panleucopenia virus.

  19. The daily lives of people with Parkinson's disease.

    PubMed

    Valcarenghi, Rafaela Vivian; Alvarez, Angela Maria; Santos, Silvana Sidney Costa; Siewert, Josiane Steil; Nunes, Simony Fabíola Lopes; Tomasi, Andrelise Viana Rosa

    2018-01-01

    To understand the daily lives of people with Parkinson's disease. Qualitative research, using as methodological and theoretical referential the Grounded Theory and Symbolic Interactionism, respectively. The in-depth interview was conducted with 30 people with Parkinson's disease. From data analysis, three themes were selected: Living with the disease - living with the treatment and changes in lifestyle; Modifying of one's job performance - revealing incapacity for work and the need to anticipate retirement and; Living with the stigma - the feeling of prejudice against the disease and the perceived limitations of the health services. Living with a chronic and non-transferable disease encompasses social, physical and cultural effects, along with the personal experiences of each unique individual. This study assists the improvement of care to people with the disease, because the care practice emerges from the interactions between the subjects.

  20. Interaction of multi-functional silver nanoparticles with living cells

    NASA Astrophysics Data System (ADS)

    Sur, Ilknur; Cam, Dilek; Kahraman, Mehmet; Baysal, Asli; Culha, Mustafa

    2010-04-01

    Silver nanoparticles (AgNPs) are widely used in household products and in medicine due to their antibacterial and to wound healing properties. In recent years, there is also an effort for their use in biomedical imaging and photothermal therapy. The primary reason behind the effort for their utility in biomedicine and therapy is their unique plasmonic properties and easy surface chemistry for a variety of functionalizations. In this study, AgNPs modified with glucose, lactose, oligonucleotides and combinations of these ligands are investigated for their cytotoxicity and cellular uptake in living non-cancer (L929) and cancer (A549) cells. It is found that the chemical nature of the ligand strongly influences the toxicity and cellular uptake into the model cells. While the lactose-and glucose-modified AgNPs enter the L929 cells at about the same rate, a significant increase in the rate of lactose-modified AgNPs into the A549 cells is observed. The binding of oligonucleotides along with the carbohydrate on the AgNP surfaces influences the differential uptake rate pattern into the cells. The cytotoxicity study with the modified AgNPs reveals that only naked AgNPs influence the viability of the A549 cells. The findings of this study may provide the key to developing effective applications in medicine such as cancer therapy.

  1. Radical SAM Enzymes in the Biosynthesis of Ribosomally Synthesized and Post-translationally Modified Peptides (RiPPs)

    PubMed Central

    Benjdia, Alhosna; Balty, Clémence; Berteau, Olivier

    2017-01-01

    Ribosomally-synthesized and post-translationally modified peptides (RiPPs) are a large and diverse family of natural products. They possess interesting biological properties such as antibiotic or anticancer activities, making them attractive for therapeutic applications. In contrast to polyketides and non-ribosomal peptides, RiPPs derive from ribosomal peptides and are post-translationally modified by diverse enzyme families. Among them, the emerging superfamily of radical SAM enzymes has been shown to play a major role. These enzymes catalyze the formation of a wide range of post-translational modifications some of them having no counterparts in living systems or synthetic chemistry. The investigation of radical SAM enzymes has not only illuminated unprecedented strategies used by living systems to tailor peptides into complex natural products but has also allowed to uncover novel RiPP families. In this review, we summarize the current knowledge on radical SAM enzymes catalyzing RiPP post-translational modifications and discuss their mechanisms and growing importance notably in the context of the human microbiota. PMID:29167789

  2. What is so important about completing lives? A critique of the modified youngest first principle of scarce resource allocation.

    PubMed

    Gamlund, Espen

    2016-04-01

    Ruth Tallman has recently offered a defense of the modified youngest first principle of scarce resource allocation [1]. According to Tallman, this principle calls for prioritizing adolescents and young adults between 15-40 years of age. In this article, I argue that Tallman's defense of the modified youngest first principle is vulnerable to important objections, and that it is thus unsuitable as a basis for allocating resources. Moreover, Tallman makes claims about the badness of death for individuals at different ages, but she lacks an account of the loss involved in dying to support her claims. To fill this gap in Tallman's account, I propose a view on the badness of death that I call 'Deprivationism'. I argue that this view explains why death is bad for those who die, and that it has some advantages over Tallman's complete lives view in the context of scarce resource allocation. Finally, I consider some objections to the relevance of Deprivationism to resource allocation, and offer my responses.

  3. [Parametabolism as Non-Specific Modifier of Supramolecular Interactions in Living Systems].

    PubMed

    Kozlov, V A; Sapozhnikov, S P; Sheptuhina, A I; Golenkov, A V

    2015-01-01

    As it became known recently, in addition to the enzyme (enzymes and/or ribozymes) in living organisms occur a large number of ordinary chemical reactions without the participation of biological catalysts. These reactions are distinguished by low speed and, as a rule, the irreversibility. For example, along with diabetes mellitus, glycation and fructosilation of proteins are observed resulted in posttranslational modification with the low- or nonfunctioning protein formation which is poorly exposed to enzymatic proteolysis and therefore accumulates in the body. In addition, the known processes such as the nonenzymatic carbomoylation, pyridoxylation and thiamiation proteins. There is a reasonable basis to believe that alcoholic injury also realized through parametabolic secondary metabolites synthesis such as acetaldehyde. At the same time, the progress in supramolecular chemistry proves that in biological objects there is another large group ofparametabolic reactions caused by the formation of supramolecular complexes. Obviously, known parameterizes interactions can modify the formation of supramolecular complexes in living objects. These processes are of considerable interest for fundamental biology and fundamental and practical medicine, but they remain unexplored due to a lack of awareness of a wide range of researchers.

  4. Relationship between risk factors and activities of daily living using modified Shah Barthel Index in stroke patients

    NASA Astrophysics Data System (ADS)

    Kusumaningsih, W.; Rachmayanti, S.; Werdhani, R. A.

    2017-08-01

    Hypertension and diabetes mellitus are the most common risk factors of stroke. The study aimed to determine the relationship between hypertension and diabetes mellitus risk factors and dependence on assistance with activities of daily living in chronic stroke patients. The study used an analytical observational cross-sectional design. The study’s sample included 44 stroke patients selected using the quota sampling method. The relationship between the variables was analyzed using the bivariate chi-squared test and multivariate logistic regression. Based on the chi-squared test, the relationship between the Modified Shah Barthel Index (MSBI) score and hypertension and diabetes mellitus as stroke risk factors, were p = 0.122 and p = 0.002, respectively. The logistic regression results suggest that hypertension and diabetes mellitus are stroke risk factors related to the MSBI score: p = 0.076 (OR 4.076; CI 95% 0.861-19.297) and p = 0.007 (OR 22.690; CI 95% 2.332-220.722), respectively. Diabetes mellitus is the most prominent risk factor of severe dependency on assistance with activities of daily living in chronic stroke patients.

  5. Selective probing of mRNA expression levels within a living cell.

    PubMed

    Nawarathna, D; Turan, T; Wickramasinghe, H Kumar

    2009-08-24

    We report on a selective and nondestructive measurement of mRNA (messenger ribonucleic acid) expression levels within a living cell. We first modify an atomic force microscope tip to create a tapered nanoscale coaxial cable. Application of an ac (alternating potential) between the inner and outer electrodes of this cable creates a dielectrophoretic force attracting mRNA molecules toward the tip-end which is pretreated with gene specific primers. We selectively extracted and analyzed both high ( approximately 2500) and extremely low (11 0) copy number mRNA from a living cell mRNA in less than 10 s.

  6. Current issues connected with usage of genetically modified crops in production of feed and livestock feeding.

    PubMed

    Kwiatek, K; Mazur, M; Sieradzki, Z

    2008-01-01

    Progress, which is brought by new advances in modern molecular biology, allowed interference in the genome of live organisms and gene manipulation. Introducing new genes to the recipient organism enables to give them new features, absent before. Continuous increase in the area of the biotech crops triggers continuous discussion about safety of genetically modified (GM) crops, including food and feed derived from them. Important issue connected with cultivation of genetically modified crops is a horizontal gene transfer and a bacterial antibiotic resistance. Discussion about safety of GM crops concerns also food allergies caused by eating genetically modified food. The problem of genetic modifications of GM crops used for livestock feeding is widely discussed, taking into account Polish feed law.

  7. In vivo monitoring of distributional transport kinetics and extravasation of quantum dots in living rat liver

    NASA Astrophysics Data System (ADS)

    Su, Cheng-Kuan; Sun, Yuh-Chang

    2013-04-01

    Although the unique optical properties of surface-modified quantum dots (QDs) have attracted wide interest in molecular biology and bioengineering, there are very few reports of their in vivo biodistribution, due to a lack of analytical techniques for characterizing the dynamic variation of QDs in living animals. In this study, we used an in vivo online monitoring system and a batch-wise elemental analytical method to investigate the biodistribution/extravasation of various surface-modified CdTeSe/ZnS (QDs) in rat liver. It is found that the surface modification dictated not only the blood retention profile but also the degree of extravasation and the clearance of extracellular QDs, making it an important variable for regulating the transfer and exchange process of QDs among three physiological compartments—bloodstream, extracellular space and Kupffer cells/hepatocytes.

  8. Multiphoton luminescent graphene quantum dots for in vivo tracking of human adipose-derived stem cells

    NASA Astrophysics Data System (ADS)

    Kim, Jin; Song, Sung Ho; Jin, Yoonhee; Park, Hyun-Ji; Yoon, Hyewon; Jeon, Seokwoo; Cho, Seung-Woo

    2016-04-01

    The applicability of graphene quantum dots (GQDs) for the in vitro and in vivo live imaging and tracking of different types of human stem cells is investigated. GQDs synthesized by the modified graphite intercalated compound method show efficient cellular uptake with improved biocompatibility and highly sensitive optical properties, indicating their feasibility as a bio-imaging probe for stem cell therapy.The applicability of graphene quantum dots (GQDs) for the in vitro and in vivo live imaging and tracking of different types of human stem cells is investigated. GQDs synthesized by the modified graphite intercalated compound method show efficient cellular uptake with improved biocompatibility and highly sensitive optical properties, indicating their feasibility as a bio-imaging probe for stem cell therapy. Electronic supplementary information (ESI) available: Additional results. See DOI: 10.1039/c6nr02143c

  9. Revitalizing genetically-modified mouse strains using frozen-thawed sperm after up to 192 h of refrigerated epididymis transportation.

    PubMed

    Moreno-Del Val, Gonzalo; Muñoz-Robledano, Patricia

    2017-10-01

    In the scientific interchange of genetically-modified mouse strains the transportation of refrigerated epididymis has several advantages over the transportation of live animals, especially with regard to the 3R (replacement, reduction and refinement) principles. The major limiting factor is the duration of the transportation. Previous reports have shown that sperm collected from transported epididymis maintained their fertility for around 72 h, but there are no published data with longer transportation times, and this window of time may be too short, especially for international shipments and where locations are not well connected. In this study live pups were born using frozen-thawed sperm after up to 192 h (8 days) of transportation, using a special in vitro fertilization design which resulted in a fertilization rate of 10.5%.

  10. Evaluation of a 20year old porcine reproductive and respiratory syndrome (PRRS) modified live vaccine (Ingelvac(®) PRRS MLV) against two recent type 2 PRRS virus isolates in South Korea.

    PubMed

    Jeong, Jiwoon; Choi, Kyuhyung; Kang, Ikjae; Park, Changhoon; Chae, Chanhee

    2016-08-30

    Type 2 porcine reproductive and respiratory syndrome (PRRS) virus (PRRSV) was first isolated in Korea in 1994. The commercial PRRS modified live vaccine (Ingelvac(®) PRRS MLV, Boehringer Ingelheim Vetmedica Inc., St. Joseph, Missouri, USA) based on type 2 PRRSV, was first licensed for use in 3- to 18-week-old pigs in Korea in 1996. The objective of the present study was to evaluate the efficacy of this 20year old commercial PRRS modified live vaccine (MLV) against two recent PRRSV isolates. Two genetically distant type 2 PRRSV strains (SNUVR150004 for lineage 1 and SNUVR150324 for lineage 5), isolated in 2015, were used as challenge virus. Regardless of the challenge virus, vaccination of pigs effectively reduced the level of viremia, the lung lesions, and of the PRRSV antigen within the lung lesions. The induction of virus-specific interferon-γ secreting cells by the PRRS vaccine produced a protective immune response, leading to the reduction of PRRSV viremia. There were no significant differences in efficacy against the two recently isolated viruses by the PRRS MLV based on virological results, immunological responses, and pathological outcomes. This study demonstrates that the PRRS MLV used in this study is still effective against recently isolated heterologous type 2 PRRSV strains even after 20 years of use in over 35 million pigs. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Evaluation of Airborne Visible/Infrared Imaging Spectrometer (AVIRIS) and Moderate Resolution Imaging Spectrometer (MODIS) measures of live fuel moisture and fuel condition in a shrubland ecosystem in southern California

    Treesearch

    D. A. Roberts; P.E. Dennison; S. Peterson; S. Sweeney; J. Rechel

    2006-01-01

    Dynamic changes in live fuel moisture (LFM) and fuel condition modify fire danger in shrublands. We investigated the empirical relationship between field-measured LFM and remotely sensed greenness and moisture measures from the Airborne Visible/Infrared Imaging Spectrometer (AVIRIS) and the Moderate Resolution Imaging Spectrometer (MODIS). Key goals were to assess the...

  12. A live attenuated H7N7 candidate vaccine virus induces neutralizing antibody that confers protection from challenge in mice, ferrets and monkeys

    USDA-ARS?s Scientific Manuscript database

    A live attenuated H7N7 candidate vaccine virus was generated by reverse genetics using the modified hemagglutinin (HA) and neuraminidase (NA) genes of HP A/Netherlands/219/03 (NL/03) (H7N7) wild-type (wt) virus and the six internal protein genes of the cold-adapted (ca) A/Ann Arbor/6/60 ca (AA ca) (...

  13. Radiation induced dechlorination of some chlorinated hydrocarbons in aqueous suspensions of various solid particles

    NASA Astrophysics Data System (ADS)

    Múčka, V.; Buňata, M.; Čuba, V.; Silber, R.; Juha, L.

    2015-07-01

    Radiation induced dechlorination of trichloroethylene (TCE) and tetrachloroethylene (PCE) in aqueous solutions containing the active carbon (AC) or cupric oxide (CuO) as the modifiers was studied. The obtained results were compared to the previously studied dechlorination of polychlorinated biphenyls (PCBs). Both modifiers were found to decrease the efficiency of dechlorination. The AC modifier acts mainly via adsorption of the aliphatic (unlike the aromatic) hydrocarbons and the CuO oxide mainly inhibits the mineralization of the perchloroethylene. The results presented in this paper will be also helpful for the studies of the impact of chlorinated hydrocarbons on the membrane permeability of living cells.

  14. Surface-modified CMOS IC electrochemical sensor array targeting single chromaffin cells for highly parallel amperometry measurements.

    PubMed

    Huang, Meng; Delacruz, Joannalyn B; Ruelas, John C; Rathore, Shailendra S; Lindau, Manfred

    2018-01-01

    Amperometry is a powerful method to record quantal release events from chromaffin cells and is widely used to assess how specific drugs modify quantal size, kinetics of release, and early fusion pore properties. Surface-modified CMOS-based electrochemical sensor arrays allow simultaneous recordings from multiple cells. A reliable, low-cost technique is presented here for efficient targeting of single cells specifically to the electrode sites. An SU-8 microwell structure is patterned on the chip surface to provide insulation for the circuitry as well as cell trapping at the electrode sites. A shifted electrode design is also incorporated to increase the flexibility of the dimension and shape of the microwells. The sensitivity of the electrodes is validated by a dopamine injection experiment. Microwells with dimensions slightly larger than the cells to be trapped ensure excellent single-cell targeting efficiency, increasing the reliability and efficiency for on-chip single-cell amperometry measurements. The surface-modified device was validated with parallel recordings of live chromaffin cells trapped in the microwells. Rapid amperometric spikes with no diffusional broadening were observed, indicating that the trapped and recorded cells were in very close contact with the electrodes. The live cell recording confirms in a single experiment that spike parameters vary significantly from cell to cell but the large number of cells recorded simultaneously provides the statistical significance.

  15. In-phase and out-of-phase axial-torsional fatigue behavior of Haynes 188 at 760 C

    NASA Technical Reports Server (NTRS)

    Kalluri, Sreeramesh; Bonacuse, Peter J.

    1991-01-01

    Isothermal, in-phase and out-of-phase axial-torsional fatigue experiments have been conducted at 760 C on uniform gage section, thin-walled tubular specimens of a wrought cobalt-base superalloy, Haynes 188. Test-control and data acquisition were accomplished with a minicomputer. Fatigue lives of the in- and out-of-phase axial-torsional fatigue tests have been estimated with four different multiaxial fatigue life prediction models that were developed primarly for predicting axial-torsional fatigue lives at room temperature. The models investigated were: (1) the von Mises equivalent strain range; (2) the Modified Multiaxiality Factor Approach; (3) the Modified Smith-Watson-Topper Parameter; and (4) the critical shear plane method of Fatemi, Socie, and Kurath. In general, life predictions by the von Mises equivalent strain range model were within a factor of 2 for a majority of the tests and the predictions by the Modified Multiaxiality Factor Approach were within a factor of 2, while predictions of the Modified Smith-Watson-Topper Parameter and of the critical shear plane method of Fatemi, Socie, and Kurath were unconservative and conservative, respectively, by up to factors of 4. In some of the specimens tested under combined axial-torsional loading conditions, fatigue cracks initiated near extensometer indentations. Two design modifications have been proposed to the thin-walled tubular specimen to overcome this problem.

  16. In vivo response of bioactive PMMA-based bone cement modified with alkoxysilane and calcium acetate.

    PubMed

    Sugino, Atsushi; Ohtsuki, Chikara; Miyazaki, Toshiki

    2008-11-01

    The use of polymethylmethacrylate (PMMA)-based bone cement is popular in orthopedics for the fixation of artificial joints with bone. However, it has a major problem with prostheses loosening because of coverage by fibrous tissue after long-term implantation. Recently, a bioactive bone cement has been developed that shows direct bonding to living bone through modification of PMMA resin with gamma-methacryloxypropyltrimethoxysilane (MPS) and calcium acetate. The cement is designed to exhibit bioactivity, through incorporation of silanol groups and calcium ions. Thus, it has the potential to form a layer of bone-like hydroxyapatite, which is essential for achieving direct bonding to living bone. This type of modification allows the cement to show spontaneous hydroxyapatite formation on its surface in a simulated body fluid after one day, and there is evidence of osteoconduction of the cement in rabbit tibia for periods of more than three weeks. However, the influence of the dissolved ions from the modified cement has not yet been clarified. Thus, the authors focused on the dissolution of the modified PMMA-based bone cement and its tissue response in muscle and bone by comparison with the behavior of non-modified PMMA-based bone cement. One week after implantation in the latissimus dorsi of a rabbit, the modified PMMA-based bone cement showed more inflammatory width than the commercial cement. However, four weeks after implantation, the inflammatory width of both cements was essentially the same. The osteoconductivity around the modified cement was higher than that for the conventional cement after four weeks implantation. These results indicate that the initial dissolution of calcium acetate from the modified cement to form the hydroxyapatite induced the acute inflammation around tissue, but also developed the osteoconductivity. It is suggested that the initial inflammation can be effective for inducing osteoconduction through a bone healing reaction when the material provides an environment that promotes bone formation.

  17. [The hospital as space and as territory].

    PubMed

    Stiefel, Friedrich; Marion-Veyron, Régis; Englebert, Jérôme

    2018-02-07

    Space is lived individually and collectively and can become a source of existential affectation, especially when the lived experience is modified by disease. The fact that the hospital is also a place of territorialisation can potentiate this affectation, with at times surprising consequences. We aim - based on a reflection about the relationship patients and clinicians establish with the territory of the hospital- to identify some psychological and existential issues at stake with regard to space as a social construction.

  18. Synergy of SOCS-1 Inhibition and Microbial-Based Cancer Vaccines

    DTIC Science & Technology

    2014-11-01

    response without causing additional risk to the patient. The goal of our proposal is to modify a live- attenuated vaccine vector based on the food -borne...response after vaccination with a live-­‐‑attenuated L. monocytogenes. Aim 3: Test the hypothesis that secretion of a SOCS-­‐‑1 small peptide ...efficient internalization of pathogens and dying cells, processing of this material into peptide antigens that are presented in the context of major

  19. Evidence of pestivirus RNA in human virus vaccines.

    PubMed Central

    Harasawa, R; Tomiyama, T

    1994-01-01

    We examined live virus vaccines against measles, mumps, and rubella for the presence of pestivirus RNA or of pestiviruses by reverse transcription PCR. Pestivirus RNA was detected in two measles-mumps-rubella combined vaccines and in two monovalent vaccines against mumps and rubella. Nucleotide sequence analysis of the PCR products indicated that a modified live vaccine strain used for immunization of cattle against bovine viral diarrhea is not responsible for the contamination of the vaccines. Images PMID:8077414

  20. Fluorescent probes for nucleic Acid visualization in fixed and live cells.

    PubMed

    Boutorine, Alexandre S; Novopashina, Darya S; Krasheninina, Olga A; Nozeret, Karine; Venyaminova, Alya G

    2013-12-11

    This review analyses the literature concerning non-fluorescent and fluorescent probes for nucleic acid imaging in fixed and living cells from the point of view of their suitability for imaging intracellular native RNA and DNA. Attention is mainly paid to fluorescent probes for fluorescence microscopy imaging. Requirements for the target-binding part and the fluorophore making up the probe are formulated. In the case of native double-stranded DNA, structure-specific and sequence-specific probes are discussed. Among the latest, three classes of dsDNA-targeting molecules are described: (i) sequence-specific peptides and proteins; (ii) triplex-forming oligonucleotides and (iii) polyamide oligo(N-methylpyrrole/N-methylimidazole) minor groove binders. Polyamides seem to be the most promising targeting agents for fluorescent probe design, however, some technical problems remain to be solved, such as the relatively low sequence specificity and the high background fluorescence inside the cells. Several examples of fluorescent probe applications for DNA imaging in fixed and living cells are cited. In the case of intracellular RNA, only modified oligonucleotides can provide such sequence-specific imaging. Several approaches for designing fluorescent probes are considered: linear fluorescent probes based on modified oligonucleotide analogs, molecular beacons, binary fluorescent probes and template-directed reactions with fluorescence probe formation, FRET donor-acceptor pairs, pyrene excimers, aptamers and others. The suitability of all these methods for living cell applications is discussed.

  1. Tetrazine-Based Cycloadditions: Application to Pretargeted Live Cell Imaging

    PubMed Central

    Devaraj, Neal K.; Weissleder, Ralph; Hilderbrand, Scott A.

    2009-01-01

    Bioorthogonal tetrazine cycloadditions have been applied to live cell labeling. Tetrazines react irreversibly with the strained dienophile norbornene forming dihydropyrazine products and dinitrogen. The reaction is high yielding, selective, and fast in aqueous media. Her2/neu receptors on live human breast cancer cells were targeted with a monoclonal antibody modified with a norbornene. Tetrazines conjugated to a near-infrared fluorochrome selectively and rapidly label the pretargeted antibody in the presence of serum. These findings indicate that this chemistry is suitable for in vitro labeling experiments, and suggests that it may prove a useful strategy for in vivo pretargeted imaging under numerous modalities. PMID:19053305

  2. Selective probing of mRNA expression levels within a living cell

    PubMed Central

    Nawarathna, D.; Turan, T.; Wickramasinghe, H. Kumar

    2009-01-01

    We report on a selective and nondestructive measurement of mRNA (messenger ribonucleic acid) expression levels within a living cell. We first modify an atomic force microscope tip to create a tapered nanoscale coaxial cable. Application of an ac (alternating potential) between the inner and outer electrodes of this cable creates a dielectrophoretic force attracting mRNA molecules toward the tip-end which is pretreated with gene specific primers. We selectively extracted and analyzed both high (∼2500) and extremely low (11¯0) copy number mRNA from a living cell mRNA in less than 10 s. PMID:19777090

  3. Vaccination-induced distemper in kinkajous.

    PubMed

    Kazacos, K R; Thacker, H L; Shivaprasad, H L; Burger, P P

    1981-12-01

    Following vaccination for distemper, using a modified live-virus vaccine developed for dogs, 2 young kinkajous (Potos flavus) developed diarrhea, then central nervous system disease. Clinical signs included myoclonus, head trembling, loss of muscular coordination, and convulsions. One kinkajou gradually recovered; the other seemed to recover, then relapsed and was euthanatized. Microscopic lesions included those of interstitial pneumonia and enteritis and multifocal lymphocytic inflammation, gliosis, spongiosis, and swollen and degenerating axons in the cerebral, cerebellar, and brain stem white matter. Similar lesions were found at all caudally, as exemplified by complete destruction of the dorsal funiculi and dorsal horns of the gray matter of the upper sacral cord segments. Eosinophilic intranuclear inclusions were seen histologically in the glia in the spinal cord, midbrain, and cerebellum, and were confirmed as canine distemper viral inclusions by the fluorescent antibody method. It was concluded that modified live canine distemper virus vaccines should be used with caution or not at all in kinkajous.

  4. Modifying the verbal expression of a child with autistic behaviors.

    PubMed

    Hargrave, E; Swisher, L

    1975-06-01

    The Bell and Howell Language Master was used in conjunction with the Monterey Language Program to modify the verbal expression of a nine-year-old boy with autistic behaviors. The goal was to train the child to correctly name up to 10 pictures presented individually. Two training modes were used. For one, the therapist spoke at the time (live voice). For the other, she presented a tape recording of her voice via a Language Master. The results suggested that the child's responses to the Language Master were as good as, if not better than, his responses to the live-voice presentations. In addition, observation indicated that he responded more readily to the Language Master presentations. His spontaneous speech was also noted by independent observers to improve in his classroom and in his home. Possible reasons for the improvement in verbal expression are considered.

  5. A time series based sequence prediction algorithm to detect activities of daily living in smart home.

    PubMed

    Marufuzzaman, M; Reaz, M B I; Ali, M A M; Rahman, L F

    2015-01-01

    The goal of smart homes is to create an intelligent environment adapting the inhabitants need and assisting the person who needs special care and safety in their daily life. This can be reached by collecting the ADL (activities of daily living) data and further analysis within existing computing elements. In this research, a very recent algorithm named sequence prediction via enhanced episode discovery (SPEED) is modified and in order to improve accuracy time component is included. The modified SPEED or M-SPEED is a sequence prediction algorithm, which modified the previous SPEED algorithm by using time duration of appliance's ON-OFF states to decide the next state. M-SPEED discovered periodic episodes of inhabitant behavior, trained it with learned episodes, and made decisions based on the obtained knowledge. The results showed that M-SPEED achieves 96.8% prediction accuracy, which is better than other time prediction algorithms like PUBS, ALZ with temporal rules and the previous SPEED. Since human behavior shows natural temporal patterns, duration times can be used to predict future events more accurately. This inhabitant activity prediction system will certainly improve the smart homes by ensuring safety and better care for elderly and handicapped people.

  6. Improved hatchability and efficient protection after in ovo vaccination with live-attenuated H7N2 and H9N2 avian influenza viruses

    PubMed Central

    2011-01-01

    Mass in ovo vaccination with live attenuated viruses is widely used in the poultry industry to protect against various infectious diseases. The worldwide outbreaks of low pathogenic and highly pathogenic avian influenza highlight the pressing need for the development of similar mass vaccination strategies against avian influenza viruses. We have previously shown that a genetically modified live attenuated avian influenza virus (LAIV) was amenable for in ovo vaccination and provided optimal protection against H5 HPAI viruses. However, in ovo vaccination against other subtypes resulted in poor hatchability and, therefore, seemed impractical. In this study, we modified the H7 and H9 hemagglutinin (HA) proteins by substituting the amino acids at the cleavage site for those found in the H6 HA subtype. We found that with this modification, a single dose in ovo vaccination of 18-day old eggs provided complete protection against homologous challenge with low pathogenic virus in ≥70% of chickens at 2 or 6 weeks post-hatching. Further, inoculation of 19-day old egg embryos with 106 EID50 of LAIVs improved hatchability to ≥90% (equivalent to unvaccinated controls) with similar levels of protection. Our findings indicate that the strategy of modifying the HA cleavage site combined with the LAIV backbone could be used for in ovo vaccination against avian influenza. Importantly, with protection conferred as early as 2 weeks post-hatching, with this strategy birds would be protected prior to or at the time of delivery to a farm or commercial operation. PMID:21255403

  7. Development of hypertrophic osteodystrophy and antibody response in a litter of vaccinated Weimaraner puppies.

    PubMed

    Harrus, S; Waner, T; Aizenberg; Safra, N; Mosenco, A; Radoshitsky, M; Bark, H

    2002-01-01

    Two different vaccination protocols were compared with regard to the development of hypertrophic osteodystrophy (HOD) (also termed metaphyseal osteopathy) and effectiveness of immunisation in a litter of 10 Weimaraner puppies. Five puppies (group 1) were vaccinated with a modified live canine parvovirus vaccine (CPV) and then two weeks later with a trivalent vaccine containing modified live canine distemper virus and adenovirus type 2 combined with a Leptospira bacterin (DHL). The CPV and DHL vaccine protocols were administered a further two times, at two-week intervals. Group 2 was vaccinated with three consecutive multivalent vaccines containing modified live canine distemper virus, canine parvovirus, parainfluenza and adenovirus type 2 combined with a Leptospira bacterin, at four-week intervals. All puppies were first vaccinated at the age of eight weeks. Three dogs in group 1 developed HOD, while all five dogs in group 2 developed HOD during the study period. Dogs in group 2 had more episodes of HOD than those in group 1. Dogs in group 1 developed higher antibody titres to canine distemper virus and parvovirus compared with those in group 2. Only two out of the 10 dogs developed protective antibody titres to parvovirus. The results of this study suggest that the two different vaccination protocols affected the pattern of appearance of HOD and immunisation in this litter of Weimaraner puppies. The results obtained and the previously reported data suggest that a larger controlled study is needed to further elucidate the effect of different vaccination protocols on HOD and immunisation in Weimaraner puppies.

  8. Perinatal Loss: The Effect on Attachment in Subsequent Pregnancies

    DTIC Science & Technology

    1989-01-01

    nurses can have the greatest impact on assisting parents to regain a state of equilibrium in their lives. Goals of Maternal -Child Health, as stated in the...negative results bring on feelings of failure and low self - esteem . There are two developmental changes which affect a women’s acceptance of the pregnancy...to the comparison group who had 2.4 living children . C. Data Collection Tool A modified version of Cranley’s Maternal Fetal Attachment Scale was

  9. The influence of the multi-basic cleavage site of the H5 hemagglutinin on the attenuation, immunogenicity and efficacy of a live attenuated influenza A h5N1 cold-adapted vaccine virus

    USDA-ARS?s Scientific Manuscript database

    A recombinant live attenuated influenza virus (LAIV) deltaH5N1 vaccine with a modified hemagglutinin (HA) and intact neuraminidase genes from A/Vietnam/1203/04 (H5N1) and the six remaining genome segments from A/Ann Arbor/6/60 (H2N2) cold-adapted (AA ca) virus was attenuated in chickens, mice and fe...

  10. Coating and dispersion of ceramic nanoparticles by UV-ozone etching assisted surface-initiated living radical polymerization.

    PubMed

    Arita, Toshihiko

    2010-10-01

    Commercially available unmodified ceramic nanoparticles (NPs) in dry powder state were surface-modified and dispersed in almost single-crystal size. The surface-initiated living radical polymerization after just UV-ozone soft etching enables one to graft polymers onto the surface of ceramic NPs and disperse them in solvents. Furthermore, a number of NPs were dispersed with single-crystal sizes. The technique developed here could be applied to almost all ceramic NPs including metal nitrides.

  11. Interaction of pollution abatement with world dynamics

    NASA Technical Reports Server (NTRS)

    Smith, G. L.

    1973-01-01

    The world dynamics model of Jay W. Forrester was modified to account for pollution abatement. In the modified model, it is assumed that as pollution increases, efforts are made to control pollution. There is a competition between food supply, material standard of living, and pollution abatement for capital, and time is required for diversion of capital toward pollution abatement. Inclusion of pollution abatement in the model drastically alters the response of the world system for the case in which depletion of natural resources is not considered. Instead of undergoing a pollution catastrophe, all system levels move more or less smoothly toward an equilibrium. A FORTRAN program listing of the modified world dynamics model is included.

  12. In vitro co-culture of human skin keratinocytes and fibroblasts on a biocompatible and biodegradable scaffold.

    PubMed

    Pajoum Shariati, Seyed Ramin; Shokrgozar, Mohammad Ali; Vossoughi, Manouchehr; Eslamifar, Ali

    2009-07-01

    Extensive full-thickness burns require replacement of both epidermis and dermis. In designing skin replacements, the goal has been to re-create this model and make a product which has both essential components. In the present study, we developed procedures for establishing confluent, stratified layers of cultured human keratinocytes on the surface of modified collagen-chitosan scaffold that contains fibroblasts. The culture methods for propagation of keratinocytes and fibroblasts isolated from human neonatal foreskin were developed. The growth and proliferation of normal human keratinocytes were evaluated in serum-free (keratinocyte growth medium) and our modified medium. Characterization of human keratinocytes was determined by using pan-keratin and anti-involucrin monoclonal antibodies. For fabrication of relevant biodegradable and biocompatible collagen-chitosan porous scaffold with improved biostability, modified method of freeze-gelation was used. In generating organotypic co-cultures, epidermal keratinocytes were plated onto the upper surface of scaffold containing embedded fibroblasts. The results showed that the growth of isolated human skin fibroblasts and keratinocytes in our modified medium was more than that in the serum-free medium. The different evaluations of collagen-chitosan scaffold showed that it is relevant to growth of cells (fibroblast and keratinocyte) and has a good flexibility in manipulation of the living skin equivalents. These findings indicate that the integration of collagen-chitosan scaffold with co-cultured keratinocyte and fibroblast in vitro provides a potential source of living skin for grafting in vivo.

  13. Parallel Prebiotic Origin of Canonical and Non-Canonical Purine Nucleosides

    NASA Astrophysics Data System (ADS)

    Becker, S.; Carell, T.

    2017-07-01

    RNA of all living organisms is highly modified. It is unclear if these non-canonical bases are ancestors of an early Earth or biological inventions. We investigated a prebiotic pathway that leads to canonical and non-canonical purine nucleosides.

  14. In Vitro Spermatogenesis of Gypsy Moth Larvae.

    ERIC Educational Resources Information Center

    Brown, Judy; Loeb, Marcia J.

    1994-01-01

    Students establish simple cell developmental cultures to observe the process of spermatogenesis, mitosis, and meiosis in living cells. Using the background information, hints for further exploration, and experimental procedures provided, teachers can easily modify this experiment to suit their students needs. (ZWH)

  15. Production of transgenic pigs over-expressing the antiviral gene Mx1.

    PubMed

    Yan, Quanmei; Yang, Huaqiang; Yang, Dongshan; Zhao, Bentian; Ouyang, Zhen; Liu, Zhaoming; Fan, Nana; Ouyang, Hongsheng; Gu, Weiwang; Lai, Liangxue

    2014-01-01

    The myxovirus resistance gene (Mx1) has a broad spectrum of antiviral activities. It is therefore an interesting candidate gene to improve disease resistance in farm animals. In this study, we report the use of somatic cell nuclear transfer (SCNT) to produce transgenic pigs over-expressing the Mx1 gene. These transgenic pigs express approximately 15-25 times more Mx1 mRNA than non-transgenic pigs, and the protein level of Mx1 was also markedly enhanced. We challenged fibroblast cells isolated from the ear skin of transgenic and control pigs with influenza A virus and classical swine fever virus (CFSV). Indirect immunofluorescence assay (IFA) revealed a profound decrease of influenza A proliferation in Mx1 transgenic cells. Growth kinetics showed an approximately 10-fold reduction of viral copies in the transgenic cells compared to non-transgenic controls. Additionally, we found that the Mx1 transgenic cells were more resistant to CSFV infection in comparison to non-transgenic cells. These results demonstrate that the Mx1 transgene can protect against viral infection in cells of transgenic pigs and indicate that the Mx1 transgene can be harnessed to develop disease-resistant pigs.

  16. Live bacterial vaccines--a review and identification of potential hazards.

    PubMed

    Detmer, Ann; Glenting, Jacob

    2006-06-23

    The use of live bacteria to induce an immune response to itself or to a carried vaccine component is an attractive vaccine strategy. Advantages of live bacterial vaccines include their mimicry of a natural infection, intrinsic adjuvant properties and their possibility to be administered orally. Derivatives of pathogenic and non-pathogenic food related bacteria are currently being evaluated as live vaccines. However, pathogenic bacteria demands for attenuation to weaken its virulence. The use of bacteria as vaccine delivery vehicles implies construction of recombinant strains that contain the gene cassette encoding the antigen. With the increased knowledge of mucosal immunity and the availability of genetic tools for heterologous gene expression the concept of live vaccine vehicles gains renewed interest. However, administration of live bacterial vaccines poses some risks. In addition, vaccination using recombinant bacteria results in the release of live recombinant organisms into nature. This places these vaccines in the debate on application of genetically modified organisms. In this review we give an overview of live bacterial vaccines on the market and describe the development of new live vaccines with a focus on attenuated bacteria and food-related lactic acid bacteria. Furthermore, we outline the safety concerns and identify the hazards associated with live bacterial vaccines and try to give some suggestions of what to consider during their development.

  17. Impact of Cropping Systems, Soil Inoculum, and Plant Species Identity on Soil Bacterial Community Structure.

    PubMed

    Ishaq, Suzanne L; Johnson, Stephen P; Miller, Zach J; Lehnhoff, Erik A; Olivo, Sarah; Yeoman, Carl J; Menalled, Fabian D

    2017-02-01

    Farming practices affect the soil microbial community, which in turn impacts crop growth and crop-weed interactions. This study assessed the modification of soil bacterial community structure by organic or conventional cropping systems, weed species identity [Amaranthus retroflexus L. (redroot pigweed) or Avena fatua L. (wild oat)], and living or sterilized inoculum. Soil from eight paired USDA-certified organic and conventional farms in north-central Montana was used as living or autoclave-sterilized inoculant into steam-pasteurized potting soil, planted with Am. retroflexus or Av. fatua and grown for two consecutive 8-week periods to condition soil nutrients and biota. Subsequently, the V3-V4 regions of the microbial 16S rRNA gene were sequenced by Illumina MiSeq. Treatments clustered significantly, with living or sterilized inoculum being the strongest delineating factor, followed by organic or conventional cropping system, then individual farm. Living inoculum-treated soil had greater species richness and was more diverse than sterile inoculum-treated soil (observed OTUs, Chao, inverse Simpson, Shannon, P < 0.001) and had more discriminant taxa delineating groups (linear discriminant analysis). Living inoculum soil contained more Chloroflexi and Acidobacteria, while the sterile inoculum soil had more Bacteroidetes, Firmicutes, Gemmatimonadetes, and Verrucomicrobia. Organically farmed inoculum-treated soil had greater species richness, more diversity (observed OTUs, Chao, Shannon, P < 0.05), and more discriminant taxa than conventionally farmed inoculum-treated soil. Cyanobacteria were higher in pots growing Am. retroflexus, regardless of inoculum type, for three of the four organic farms. Results highlight the potential of cropping systems and species identity to modify soil bacterial communities, subsequently modifying plant growth and crop-weed competition.

  18. Chemical and Biological Tools for the Preparation of Modified Histone Proteins

    PubMed Central

    Howard, Cecil J.; Yu, Ruixuan R.; Gardner, Miranda L.; Shimko, John C.; Ottesen, Jennifer J.

    2016-01-01

    Eukaryotic chromatin is a complex and dynamic system in which the DNA double helix is organized and protected by interactions with histone proteins. This system is regulated through, a large network of dynamic post-translational modifications (PTMs) exists to ensure proper gene transcription, DNA repair, and other processes involving DNA. Homogenous protein samples with precisely characterized modification sites are necessary to better understand the functions of modified histone proteins. Here, we discuss sets of chemical and biological tools that have been developed for the preparation of modified histones, with a focus on the appropriate choice of tool for a given target. We start with genetic approaches for the creation of modified histones, including the incorporation of genetic mimics of histone modifications, chemical installation of modification analogs, and the use of the expanded genetic code to incorporate modified amino acids. Additionally, we will cover the chemical ligation techniques that have been invaluable in the generation of complex modified histones that are indistinguishable from the natural counterparts. Finally, we will end with a prospectus on future directions of synthetic chromatin in living systems. PMID:25863817

  19. Developing live bacterial vaccines by selecting resistance to antibacterials

    USDA-ARS?s Scientific Manuscript database

    Four chemicals were used in this study to modify bacterial isolates through chemical-resistance strategy. All bacteria were able to develop high resistance to gossypol. However, none of the gossypol-resistant isolate was attenuated. Although majority of the proflavine hemisulfate-resistant isolates ...

  20. Vibrations that live long and prosper

    NASA Astrophysics Data System (ADS)

    Utz, Arthur L.

    2018-06-01

    Molecular vibrations can be highly effective promoters of gas-phase chemistry. Now, measurements show that excited vibrational states can survive on metal surfaces far longer than expected — reshaping our understanding of how vibrational excitation might also promote or modify heterogeneously catalysed chemistry on metals.

  1. Reduction of Radon Progeny in Indoor Air.

    DTIC Science & Technology

    1986-03-01

    arises from indoor radon is due * 4 to inhalation of the short-lived radon daughters Ra-A, Ra-B, and Ra-C. These decay products are formed from the alpha...concentrations of radon daughters 40 ’ in an air sample from the gross alpha counting of a filter 50 ’ in accordance with the modified Tsivoglou method. 60 ’ 70...8217 The modified Tsivoglou method may be found in " Measurement 80 ’ of Radon Daughters in Air," Health Physics, 23, : pp7S3-789 90 ’ (19𔃼). 95 100 The

  2. Estimation of maternal and neonatal mortality at the subnational level in Liberia.

    PubMed

    Moseson, Heidi; Massaquoi, Moses; Bawo, Luke; Birch, Linda; Dahn, Bernice; Zolia, Yah; Barreix, Maria; Gerdts, Caitlin

    2014-11-01

    To establish representative local-area baseline estimates of maternal and neonatal mortality using a novel adjusted sisterhood method. The status of maternal and neonatal health in Bomi County, Liberia, was investigated in June 2013 using a population-based survey (n=1985). The standard direct sisterhood method was modified to account for place and time of maternal death to enable calculation of subnational estimates. The modified method of measuring maternal mortality successfully enabled the calculation of area-specific estimates. Of 71 reported deaths of sisters, 18 (25.4%) were due to pregnancy-related causes and had occurred in the past 3 years in Bomi County. The estimated maternal mortality ratio was 890 maternal deaths for every 100 000 live births (95% CI, 497-1301]. The neonatal mortality rate was estimated to be 47 deaths for every 1000 live births (95% CI, 42-52). In total, 322 (16.9%) of 1900 women with accurate age data reported having had a stillbirth. The modified direct sisterhood method may be useful to other countries seeking a more regionally nuanced understanding of areas in which neonatal and maternal mortality levels still need to be reduced to meet Millennium Development Goals. Copyright © 2014 International Federation of Gynecology and Obstetrics. Published by Elsevier Ireland Ltd. All rights reserved.

  3. Chitosan coated carbon fiber microelectrode for selective in vivo detection of neurotransmitters in live zebrafish embryos

    PubMed Central

    Özel, Rıfat Emrah; Wallace, Kenneth N.; Andreescu, Silvana

    2011-01-01

    We report the development of a chitosan modified carbon fiber microelectrode for in vivo detection of serotonin. We find that chitosan has the ability to reject physiological levels of ascorbic acid interferences and facilitate selective and sensitive detection of in vivo levels of serotonin, a common catecholamine neurotransmitter. Presence of chitosan on the microelectrode surface was investigated using scanning electron microscopy (SEM) and cyclic voltammetry (CV). The electrode was characterized using differential pulse voltammetry (DPV). A detection limit of 1.6 nM serotonin with a sensitivity of 5.12 nA/µM, a linear range from 2 to 100 nM and a reproducibility of 6.5 % for n=6 electrodes were obtained. Chitosan modified microelectrodes selectively measure serotonin in presence of physiological levels of ascorbic acid. In vivo measurements were performed to measure concentration of serotonin in the live embryonic zebrafish intestine. The sensor quantifies in vivo intestinal levels of serotonin while successfully rejecting ascorbic acid interferences. We demonstrate that chitosan can be used as an effective coating to reject ascorbic acid interferences at carbon fiber microelectrodes, as an alternative to Nafion, and that chitosan modified microelectrodes are reliable tools for in vivo monitoring of changes in neurotransmitter levels. PMID:21601035

  4. The effect of the serum corona on interactions between a single nano-object and a living cell

    NASA Astrophysics Data System (ADS)

    Dror, Yael; Sorkin, Raya; Brand, Guy; Boubriak, Olga; Urban, Jill; Klein, Jacob

    2017-04-01

    Nanoparticles (NPs) which enter physiological fluids are rapidly coated by proteins, forming a so-called corona which may strongly modify their interaction with tissues and cells relative to the bare NPs. In this work the interactions between a living cell and a nano-object, and in particular the effect on this of the adsorption of serum proteins, are directly examined by measuring the forces arising as an Atomic Force Microscope tip (diameter 20 nm) - simulating a nano-object - approaches and contacts a cell. We find that the presence of a serum protein corona on the tip strongly modifies the interaction as indicated by pronounced increase in the indentation, hysteresis and work of adhesion compared to a bare tip. Classically one expects an AFM tip interacting with a cell surface to be repelled due to cell elastic distortion, offset by tip-cell adhesion, and indeed such a model fits the bare-tip/cell interaction, in agreement with earlier work. However, the force plots obtained with serum-modified tips are very different, indicating that the cell is much more compliant to the approaching tip. The insights obtained in this work may promote better design of NPs for drug delivery and other nano-medical applications.

  5. The effect of the serum corona on interactions between a single nano-object and a living cell

    PubMed Central

    Dror, Yael; Sorkin, Raya; Brand, Guy; Boubriak, Olga; Urban, Jill; Klein, Jacob

    2017-01-01

    Nanoparticles (NPs) which enter physiological fluids are rapidly coated by proteins, forming a so-called corona which may strongly modify their interaction with tissues and cells relative to the bare NPs. In this work the interactions between a living cell and a nano-object, and in particular the effect on this of the adsorption of serum proteins, are directly examined by measuring the forces arising as an Atomic Force Microscope tip (diameter 20 nm) - simulating a nano-object - approaches and contacts a cell. We find that the presence of a serum protein corona on the tip strongly modifies the interaction as indicated by pronounced increase in the indentation, hysteresis and work of adhesion compared to a bare tip. Classically one expects an AFM tip interacting with a cell surface to be repelled due to cell elastic distortion, offset by tip-cell adhesion, and indeed such a model fits the bare-tip/cell interaction, in agreement with earlier work. However, the force plots obtained with serum-modified tips are very different, indicating that the cell is much more compliant to the approaching tip. The insights obtained in this work may promote better design of NPs for drug delivery and other nano-medical applications. PMID:28383528

  6. Faecal shedding of canine parvovirus after modified-live vaccination in healthy adult dogs.

    PubMed

    Freisl, M; Speck, S; Truyen, U; Reese, S; Proksch, A-L; Hartmann, K

    2017-01-01

    Since little is known about the persistence and faecal shedding of canine parvovirus (CPV) in dogs after modified-live vaccination, diagnostic tests for CPV can be difficult to interpret in the post-vaccination period. The primary aim of this study was to determine the incidence, duration and extent of CPV vaccine virus shedding in adult dogs and to investigate related factors, including the presence of protective antibodies, increase in anti-CPV antibody titres and development of any gastrointestinal side-effects. A secondary objective was to assess prevalence of CPV field virus shedding in clinically healthy dogs due to subclinical infections. One hundred adult, healthy privately owned dogs were vaccinated with a commercial CPV-2 modified-live vaccine (MLV). Faeces were tested for the presence of CPV DNA on days 0 (prior to vaccination), 3, 7, 14, 21 and 28 by quantitative real-time PCR. Pre- and post-vaccination serum titres were determined by haemagglutination inhibition on days 0, 7 and 28. Transient excretion of CPV DNA was detected in 2.0% of dogs before vaccination. About one quarter of dogs (23.0%) shed CPV DNA during the post-vaccination period, but field and vaccine virus differentiation by VP2 gene sequencing was only successful in few samples. Faecal CPV excretion occurred despite protective serum antibody titres. Post-vaccination CPV shedding was not related to adequate antibody response after vaccination or to the occurrence of gastrointestinal side-effects. Despite individual differences, CPV DNA was detectable for up to 28 days after vaccination, although the faecal CPV DNA load in these clinically healthy dogs was very low. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Effect of the Modified Live Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Vaccine on European and North American PRRSV Shedding in Semen from Infected Boars ▿

    PubMed Central

    Han, Kiwon; Seo, Hwi Won; Shin, Jeoung Hwa; Oh, Yeonsu; Kang, Ikjae; Park, Changhoon; Chae, Chanhee

    2011-01-01

    The objective of the present study was to compare the effects of the modified live porcine reproductive and respiratory syndrome virus (PRRSV) vaccine (Ingelvac PRRS MLV; Boehringer Ingelheim Animal Health, St. Joseph, MO) on European and North American PRRSV shedding in the semen of experimentally infected boars. The boars were randomly divided into six groups. Vaccinated boars shed the North American PRRSV at the rate of 100.1 to 101.0 viral genome copies per ml and 3.63 to 101.1 50% tissue culture infective doses (TCID50)/ml, respectively, in semen, whereas nonvaccinated boars shed the North American PRRSV at the rate of 100.2 to 104.7 viral genome copies per ml and 1.14 to 103.07 TCID50/ml, respectively, in semen. Vaccinated boars shed the European PRRSV at the rate of 100.1 to 104.57 viral genome copies per ml and 1.66 to 103.10 TCID50/ml, respectively, in semen, whereas nonvaccinated boars shed the European PRRSV at the rate of 100.3 to 105.14 viral genome copies per ml and 1.69 to 103.17 TCID50/ml, respectively, in semen. The number of genomic copies of the European PRRSV in semen samples was not significantly different between vaccinated and nonvaccinated challenged European PRRSV boars. The present study demonstrated that boar vaccination using commercial modified live PRRSV vaccine was able to decrease subsequent shedding of North American PRRSV in semen after challenge but was unable to decrease shedding of European PRRSV in semen after challenge. PMID:21832096

  8. Influence of social and material individual and area deprivation on suicide mortality among 2.7 million Canadians: a prospective study.

    PubMed

    Burrows, Stephanie; Auger, Nathalie; Gamache, Philippe; St-Laurent, Danielle; Hamel, Denis

    2011-07-19

    Few studies have investigated how area-level deprivation influences the relationship between individual disadvantage and suicide mortality. The aim of this study was to examine individual measures of material and social disadvantage in relation to suicide mortality in Canada and to determine whether these relationships were modified by area deprivation. Using the 1991-2001 Canadian Census Mortality Follow-up Study cohort (N = 2,685,400), measures of individual social (civil status, family structure, living alone) and material (education, income, employment) disadvantage were entered into Cox proportional hazard models to calculate hazard ratios (HR) and 95% confidence intervals (CI) for male and female suicide mortality. Two indices of area deprivation were computed - one capturing social, and the other material, dimensions - and models were run separately for high versus low deprivation. After accounting for individual and area characteristics, individual social and material disadvantage were associated with higher suicide mortality, especially for individuals not employed, not married, with low education and low income. Associations between social and material area deprivation and suicide mortality largely disappeared upon adjustment for individual-level disadvantage. In stratified analyses, suicide risk was greater for low income females in socially deprived areas and males living alone in materially deprived areas, and there was no evidence of other modifying effects of area deprivation. Individual disadvantage was associated with suicide mortality, particularly for males. With some exceptions, there was little evidence that area deprivation modified the influence of individual disadvantage on suicide risk. Prevention strategies should primarily focus on individuals who are unemployed or out of the labour force, and have low education or income. Individuals with low income or who are living alone in deprived areas should also be targeted.

  9. Does neighbourhood composition modify the association between acculturation and unhealthy dietary behaviours?

    PubMed

    Zhang, Donglan; van Meijgaard, Jeroen; Shi, Lu; Cole, Brian; Fielding, Jonathan

    2015-08-01

    Studies have shown that immigrants' acculturation is associated with numerous unhealthy behaviours. Yet, the role of environmental factors in modifying the effect of acculturation on health behaviours has received little attention. This study aims to create a more nuanced understanding of the health effects of acculturation by examining how neighbourhood immigrant composition modifies the association between individuals' eating patterns and acculturation. Cross-sectional Data from Los Angeles County Health Survey 2007 adult sample were linked to data on retail food establishments and US Census 2000 neighbourhood characteristics. Acculturation was measured by language spoken at home and years stayed in the US. Eating fast food more than once per week and eating zero serving of fruit or vegetables during the previous day were used as proxy indicators for unhealthy dietary behaviour. Multilevel logistic regression models were performed in the full sample and in the sample with only Latino adults. Immigrants' lack of acculturation and living in a neighbourhood with a high percentage immigrants were associated with healthier dietary behaviour. We also identified that lack of acculturation conveyed a significantly stronger protective effect on regular fast-food consumption for immigrants living in neighbourhoods with higher percentage immigrants (OR: 0.34, 95% CI: 0.12 to 0.93). Among immigrants in Los Angeles County, living in a neighbourhood with a high density of other immigrants attenuates the negative effects of acculturation on healthy eating behaviours. Healthy eating promotion efforts should build on this protective effect in outreach to acculturating immigrant communities. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

  10. Protective effects against abortion and fetal infection following exposure to bovine viral diarrhea virus and bovine herpesvirus 1 during pregnancy in beef heifers that received two doses of a multivalent modified-live virus vaccine prior to breeding.

    PubMed

    Givens, M Daniel; Marley, M Shonda D; Jones, Craig A; Ensley, Douglas T; Galik, Patricia K; Zhang, Yijing; Riddell, Kay P; Joiner, Kellye S; Brodersen, Bruce W; Rodning, Soren P

    2012-08-15

    To determine whether administration of 2 doses of a multivalent, modified-live virus vaccine prior to breeding of heifers would provide protection against abortion and fetal infection following exposure of pregnant heifers to cattle persistently infected (PI) with bovine viral diarrhea virus (BVDV) and cattle with acute bovine herpesvirus 1 (BHV1) infection. Randomized controlled clinical trial. 33 crossbred beef heifers, 3 steers, 6 bulls, and 25 calves. 20 of 22 vaccinated and 10 of 11 unvaccinated heifers became pregnant and were commingled with 3 steers PI with BVDV type 1a, 1b, or 2 for 56 days beginning 102 days after the second vaccination (administered 30 days after the first vaccination). Eighty days following removal of BVDV-PI steers, heifers were commingled with 3 bulls with acute BHV1 infection for 14 days. After BVDV exposure, 1 fetus (not evaluated) was aborted by a vaccinated heifer; BVDV was detected in 0 of 19 calves from vaccinated heifers and in all 4 fetuses (aborted after BHV1 exposure) and 6 calves from unvaccinated heifers. Bovine herpesvirus 1 was not detected in any fetus or calf and associated fetal membranes in either treatment group. Vaccinated heifers had longer gestation periods and calves with greater birth weights, weaning weights, average daily gains, and market value at weaning, compared with those for calves born to unvaccinated heifers. Prebreeding administration of a modified-live virus vaccine to heifers resulted in fewer abortions and BVDV-PI offspring and improved growth and increased market value of weaned calves.

  11. Control and eradication of porcine reproductive and respiratory syndrome virus type 2 using a modified-live type 2 vaccine in combination with a load, close, homogenise model: an area elimination study.

    PubMed

    Rathkjen, Poul H; Dall, Johannes

    2017-01-05

    Porcine reproductive and respiratory syndrome virus (PRRSV) causes significant animal and economic losses worldwide. The infection is difficult to control and PRRSV elimination at local level requires coordinated intervention among multiple farms. This case study describes a successful elimination of PRRSV from all 12 herds on the Horne Peninsula, Denmark, using a combination of load, close, homogenise (LCH) using PRRSV type 2 modified-live vaccine, optimised pig flow, and'10 Golden Rules' (10GR) for biosecurity management. To the authors' knowledge, this is the first successful European PRRSV area elimination project documented in detail. The PRRSV type 2 modified-live vaccine was used as part of the LCH method in breeding herds. Complete or partial depopulation was performed in some infected herds. A simplified biosecurity protocol (10GR) based on the McREBEL™ system of pig flow management, was employed in all herds and at all times throughout the study. At study commencement, all herds were infected with PRRSV, and most were actively shedding virus. In just over 18 months, all 12 herds on the Horne Peninsula were confirmed to be PRRSV negative by polymerase chain reaction testing and negative for antibodies against PRRSV by enzyme-linked immunosorbent assay testing. All herds were subsequently obtained 'Specific Pathogen Free' status for PRRSV. This study provides compelling evidence suggesting that an area elimination plan combining LCH with PRRSV type 2 vaccination, optimised pig flow, and 10GR for biosecurity management can effectively eliminate PRRSV from a geographic area. Additionally this study confirms the value of a previously unpublished, simplified alternative to the McREBEL system for controlling PRRSV.

  12. Compatibility of a bivalent modified-live vaccine against Bordetella bronchiseptica and CPiV, and a trivalent modified-live vaccine against CPV, CDV and CAV-2.

    PubMed

    Jacobs, A A C; Bergman, J G H E; Theelen, R P H; Jaspers, R; Helps, J M; Horspool, L J I; Paul, G

    2007-01-13

    Eight puppies (group 1) were vaccinated once with a bivalent modified-live vaccine against infectious tracheobronchitis by the intranasal route and at the same time with an injectable trivalent vaccine against canine parvovirus, canine distemper virus and canine adenovirus; a second group of eight puppies (group 2) was vaccinated only with the intranasal bivalent vaccine, and a further eight puppies (group 3) were vaccinated only with the injectable trivalent vaccine. Three weeks later they were all challenged with wildtype Bordetella bronchiseptica and canine parainfluenza virus by the aerosol route, and their antibody responses to the five vaccine organisms were determined. Oronasal swabs were taken regularly before and after the challenge for the isolation of bacteria and viruses, and the puppies were observed for clinical signs for three weeks after the challenge. There were no significant differences in the puppies' titres against canine parvovirus, canine distemper virus and canine adenovirus type 2 between the groups vaccinated with or without the bivalent intranasal vaccine. After the challenge the mean clinical scores of the two groups vaccinated with the intranasal vaccine were nearly 90 per cent lower (P=0.001) than the mean score of the group vaccinated with only the trivalent injectable vaccine, and the puppies in this group all became culture-positive for B bronchiseptica and canine parainfluenza virus. There were only small differences between the rates of isolation of B bronchiseptica from groups 1, 2 and 3, but significantly lower yields of canine parainfluenza virus were isolated from groups 1 and 2 than from group 3.

  13. Immunogenicity of a low-passage, high-titer modified live canine parvovirus vaccine in pups with maternally derived antibodies.

    PubMed

    Hoare, C M; DeBouck, P; Wiseman, A

    1997-02-01

    The study evaluated the ability of a low-passage, high-titer modified live canine parvovirus (CPV) vaccine to produce seroconversion in pups with maternally derived hemagglutination inhibition (HI) titers ranging from < 8 to < or = 256. The vaccine's low-passage CPV strain was less attenuated and therefore more infective than conventional modified live CPV strains in order to overcome relatively greater levels of maternally derived antibodies, the principal cause of CPV vaccine failures in pups. To assess vaccine performance under field conditions, healthy pups presented at five private veterinary clinics were used as test animals. A single dose of vaccine was given to 59 pups at 12 weeks of age (Group A). To accommodate the protocol of clinics where earlier CPV vaccination was practiced, 87 other pups were vaccinated with two doses, the first at 8-10 weeks of age, and the second at 12 weeks of age (Group B). Geometric mean HI titers were measured for blood samples obtained at the time of vaccination and at 14 weeks of age. Seroconversion was considered to have occurred if pups developed a fourfold or greater increase in HI titer to a level > or = 64. Of the 59 pups in Group A, 100% seroconverted following the single vaccine dose at 12 weeks of age. Of the 87 Group B pups, 82 (94.3%) seroconverted following either of the two vaccine doses. A geometric mean HI titer of 4828 was measured for Group A, and a geometric mean HI titer of 2028 was measured for Group B. An overall seroconversion rate of 96.5% was achieved in pups with maternally derived HI titers < or = 256.

  14. Association of selenium status and blood glutathione concentrations in blacks and whites

    PubMed Central

    Richie, John P.; Muscat, Joshua E.; Ellison, Irina; Calcagnotto, Ana; Kleinman, Wayne; El-Bayoumy, Karam

    2011-01-01

    Selenium deficiency has been linked with increased cancer risk and, in some studies, selenium supplementation was protective against certain cancers. Previous studies suggest that selenium chemoprevention may involve reduced oxidative stress through enhanced glutathione (GSH). Our objectives were to examine the relationships between selenium and GSH in blood and modifying effects of race and sex in free living adults and individuals supplemented with selenium. Plasma selenium concentrations and free and bound GSH concentrations and γ-glutamyl cysteine ligase (GCL) activity in blood were measured in 336 healthy adults, (161 blacks, 175 whites). Plasma selenium and blood GSH were also measured in 36 healthy men from our previously conducted placebo-controlled trial of selenium-enriched yeast (247 μg/day for 9 months). In free-living adults, selenium concentrations were associated with increased blood GSH concentration and GCL activity (P<0.05). Further, selenium was significantly higher in whites than in blacks (P<0.01). After 9 months of supplementation, plasma selenium was increased 114% in whites and 50% in blacks (P<0.05) and blood GSH was increased 35% in whites (P<0.05) but was unchanged in blacks. These results indicate a direct association between selenium and GSH in blood of both free-living and selenium-supplemented individuals, with race being an important modifying factor. PMID:21462082

  15. State-Level Policies and Psychiatric Morbidity In Lesbian, Gay, and Bisexual Populations

    PubMed Central

    Keyes, Katherine M.; Hasin, Deborah S.

    2009-01-01

    Objectives. We investigated the modifying effect of state-level policies on the association between lesbian, gay, or bisexual status and the prevalence of psychiatric disorders. Methods. Data were from wave 2 of the National Epidemiologic Survey on Alcohol and Related Conditions (NESARC), a nationally representative study of noninstitutionalized US adults (N = 34 653). States were coded for policies extending protections against hate crimes and employment discrimination based on sexual orientation. Results. Compared with living in states with policies extending protections, living in states without these policies predicted a significantly stronger association between lesbian, gay, or bisexual status and psychiatric disorders in the past 12 months, including generalized anxiety disorder (F = 3.87; df = 2; P = .02), post-traumatic stress disorder (F = 3.42; df = 2; P = .04), and dysthymia (F = 5.20; df = 2; P = .02). Living in states with policies that did not extend protections also predicted a stronger relation between lesbian, gay, or bisexual status and psychiatric comorbidity (F = 2.47; df = 2; P = .04). Conclusions. State-level protective policies modify the effect of lesbian, gay, or bisexual status on psychiatric disorders. Policies that reduce discrimination against gays and lesbians are urgently needed to protect the health and well-being of this population. PMID:19833997

  16. Genetically modified organisms and visceral leishmaniasis.

    PubMed

    Chhajer, Rudra; Ali, Nahid

    2014-01-01

    Vaccination is the most effective method of preventing infectious diseases. Since the eradication of small pox in 1976, many other potentially life compromising if not threatening diseases have been dealt with subsequently. This event was a major leap not only in the scientific world already burdened with many diseases but also in the mindset of the common man who became more receptive to novel treatment options. Among the many protozoan diseases, the leishmaniases have emerged as one of the largest parasite killers of the world, second only to malaria. There are three types of leishmaniasis namely cutaneous (CL), mucocutaneous (ML), and visceral (VL), caused by a group of more than 20 species of Leishmania parasites. Visceral leishmaniasis, also known as kala-azar is the most severe form and almost fatal if untreated. Since the first attempts at leishmanization, we have killed parasite vaccines, subunit protein, or DNA vaccines, and now we have live recombinant carrier vaccines and live attenuated parasite vaccines under various stages of development. Although some research has shown promising results, many more potential genes need to be evaluated as live attenuated vaccine candidates. This mini-review attempts to summarize the success and failures of genetically modified organisms used in vaccination against some of major parasitic diseases for their application in leishmaniasis.

  17. Communicating cancer-prevention information.

    PubMed

    Brody, J E

    1992-01-01

    The message to the public regarding cancer prevention should emphasize moderation and evolutionary change in living habits rather than revolutions and focus on total life-style not just individual factors. It is essential to keep people focused on the modifiable factors that can really make a difference. Currently, people worry about minutiae, ie, possible substances in the environment that might cause some cancers, rather than the main, more clearly defined factors, eg, smoking, diet, and sedentary living. The media must play a major role in cancer prevention, assisted by rational, clear-speaking professionals.

  18. Mobile Learning

    ERIC Educational Resources Information Center

    Erickson, Paul W.

    2011-01-01

    Children are doing less physical activity and sitting more than they did 20 years ago; exploring physical abilities used to be the norm. But societal changes have modified people's living conditions--children take part in "adult-initiated" activities; they sit statically, influenced by technology gaming/information; and over-cautious parents often…

  19. Vaccine Mediated Protection Against Zika Virus-Induced Congenital Disease.

    PubMed

    Richner, Justin M; Jagger, Brett W; Shan, Chao; Fontes, Camila R; Dowd, Kimberly A; Cao, Bin; Himansu, Sunny; Caine, Elizabeth A; Nunes, Bruno T D; Medeiros, Daniele B A; Muruato, Antonio E; Foreman, Bryant M; Luo, Huanle; Wang, Tian; Barrett, Alan D; Weaver, Scott C; Vasconcelos, Pedro F C; Rossi, Shannan L; Ciaramella, Giuseppe; Mysorekar, Indira U; Pierson, Theodore C; Shi, Pei-Yong; Diamond, Michael S

    2017-07-13

    The emergence of Zika virus (ZIKV) and its association with congenital malformations has prompted the rapid development of vaccines. Although efficacy with multiple viral vaccine platforms has been established in animals, no study has addressed protection during pregnancy. We tested in mice two vaccine platforms, a lipid nanoparticle-encapsulated modified mRNA vaccine encoding ZIKV prM and E genes and a live-attenuated ZIKV strain encoding an NS1 protein without glycosylation, for their ability to protect against transmission to the fetus. Vaccinated dams challenged with a heterologous ZIKV strain at embryo day 6 (E6) and evaluated at E13 showed markedly diminished levels of viral RNA in maternal, placental, and fetal tissues, which resulted in protection against placental damage and fetal demise. As modified mRNA and live-attenuated vaccine platforms can restrict in utero transmission of ZIKV in mice, their further development in humans to prevent congenital ZIKV syndrome is warranted. Copyright © 2017 Elsevier Inc. All rights reserved.

  20. Three-year duration of immunity in dogs following vaccination against canine adenovirus type-1, canine parvovirus, and canine distemper virus.

    PubMed

    Gore, Thomas C; Lakshmanan, Nallakannu; Duncan, Karen L; Coyne, Michael J; Lum, Melissa A; Sterner, Frank J

    2005-01-01

    A challenge-of-immunity study was conducted to demonstrate immunity in dogs 3 years after their second vaccination with a new multivalent, modified-live vaccine containing canine adenovirus type 2 (CAV-2), canine parvovirus (CPV), and canine distemper virus (CDV). Twenty-three seronegative pups were vaccinated at 7 and 11 weeks of age. Eighteen seronegative pups, randomized into groups of six dogs, served as challenge controls. Dogs were kept in strict isolation for 3 years following the vaccination and then challenged sequentially with virulent canine adenovirus type 1 (CAV-1), CPV, and CDV. For each viral challenge, a separate group of six control dogs was also challenged. Clinical signs of CAV-1, CPV, and CDV infections were prevented in 100% of vaccinated dogs, demonstrating that the multivalent, modified-live test vaccine provided protection against virulent CAV-1, CPV, and CDV challenge in dogs 7 weeks of age or older for a minimum of 3 years following second vaccination.

  1. Power Calculations and Placebo Effect for Future Clinical Trials in Progressive Supranuclear Palsy

    PubMed Central

    Stamelou, Maria; Schöpe, Jakob; Wagenpfeil, Stefan; Ser, Teodoro Del; Bang, Jee; Lobach, Iryna Y.; Luong, Phi; Respondek, Gesine; Oertel, Wolfgang H.; Boxer, Adam L.; Höglinger, Günter U.

    2016-01-01

    Background Two recent randomized, placebo-controlled trials of putative disease-modifying agents (davunetide, tideglusib) in progressive supranuclear palsy (PSP) failed to show efficacy, but generated data relevant for future trials. Methods We provide sample size calculations based on data collected in 187 PSP patients assigned to placebo in these trials. A placebo effect was calculated. Results The total PSP-Rating Scale required the least number of patients per group (N = 51) to detect a 50% change in the 1-year progression and 39 when including patients with ≤ 5 years disease duration. The Schwab and England Activities of Daily Living required 70 patients per group and was highly correlated with the PSP-Rating Scale. A placebo effect was not detected in these scales. Conclusions We propose the 1-year PSP-Rating Scale score change as the single primary readout in clinical neuroprotective or disease-modifying trials. The Schwab and England Activities of Daily Living could be used as a secondary outcome. PMID:26948290

  2. Serologic response of domestic ferrets (Mustela putorius furo) to canine distemper and rabies virus vaccines.

    PubMed

    Hoover, J P; Baldwin, C A; Rupprecht, C E

    1989-01-15

    Nine unrelated 12-week-old naive domestic ferrets (Mustela putorius furo) were used to evaluate the serologic responses to commercial canine distemper virus (CDV) and rabies virus (RV) vaccines. Five of the ferrets (group 1) were inoculated 3 times at 2-week intervals with a multivalent modified-live virus vaccine of canine cell-line origin, containing CDV and an inactivated RV vaccine. Four of the ferrets (group 2) were inoculated once with the multivalent modified-live virus vaccine containing CDV and were not inoculated with the RV vaccine. Both group-1 and group-2 ferrets seroconverted to the CDV component of the vaccine. Group-1 ferrets also seroconverted after RV vaccination and maintained serum antibody titers to both CDV and RV for at least 7 months. Domestic ferret sera were found to have IgG epitopes similar to sera of domestic dogs and cats. Domestic ferret sera did not contain antibodies to feline coronavirus or FeLV antigens.

  3. Modification of selected South Carolina bridge-scour envelope curves

    USGS Publications Warehouse

    Benedict, Stephen T.; Caldwell, Andral W.

    2012-01-01

    Historic scour was investigated at 231 bridges in the Piedmont and Coastal Plain physiographic provinces of South Carolina by the U.S. Geological Survey in cooperation with the South Carolina Department of Transportation. These investigations led to the development of field-derived envelope curves that provided supplementary tools to assess the potential for scour at bridges in South Carolina for selected scour components that included clear-water abutment, contraction, and pier scour, and live-bed pier and contraction scour. The envelope curves consist of a single curve with one explanatory variable encompassing all of the measured field data for the respective scour components. In the current investigation, the clear-water abutment-scour and live-bed contraction-scour envelope curves were modified to include a family of curves that utilized two explanatory variables, providing a means to further refine the assessment of scour potential for those specific scour components. The modified envelope curves and guidance for their application are presented in this report.

  4. Post-vaccinal distemper encephalitis in two Border Collie cross littermates.

    PubMed

    Fairley, R A; Knesl, O; Pesavento, P A; Elias, B C

    2015-03-01

    One 4.5-month-old male Border Collie cross presented with aggression and seizures in October 2006. A 16-month-old, female, spayed Border Collie cross presented with hypersalivation and a dropped jaw and rapidly became stuporous in September 2007. The dogs were littermates and developed acute neurological signs 5 and 27 days, respectively, after vaccination with different modified live vaccines containing canine distemper virus. Sections of brain in both dogs showed evidence of encephalitis mainly centred on the grey matter of brainstem nuclei, where there was extensive and intense parenchymal and perivascular infiltration of histiocytes and lymphocytes. Intra-nuclear and intra-cytoplasmic inclusions typical of distemper were plentiful and there was abundant labelling for canine distemper virus using immunohistochemistry. Post-vaccinal canine distemper. Post-vaccinal canine distemper has mainly been attributed to virulent vaccine virus, but it may also occur in dogs whose immunologic nature makes them susceptible to disease induced by a modified-live vaccine virus that is safe and protective for most dogs.

  5. [Research on the Kosmos biosatellites].

    PubMed

    Il'in, E A

    1984-01-01

    In the last decade the USSR has launched six biosatellites of the Cosmos series. The duration of the first flight was 6 days and of the five subsequent flights 18 to 21 days. The major goals of the flight studies were: investigation of adaptation of living systems to weightlessness, identification of the modifying effect of weightlessness on radiosensitivity, and detection of the biological effect of artificial gravity. The examinations were performed on 37 biological species, with most of them on rats. The exposure to weightlessness gave rise to moderate stress reactions and specific changes, particularly in the musculo-skeletal system (muscle atrophy, reduced bone strength, etc). Artificial gravity of 1 g generated inflight helped maintain the normal function of most physiological systems. The exposure of mammals (rats) to 137Ce irradiation did not reveal a modifying effect of weightlessness on radiation sickness. Distinct manifestations of the effects of weightlessness on intracellular processes were not observed. Dissimilar results were obtained with respect to the growth and development of living organisms in weightlessness.

  6. Place of Residence Modifies the Association Between Acculturation and Dietary Tools Knowledge Among Latina WIC Participants: A Multi-State Study

    PubMed Central

    Song, David Sheng; Taylor, Christopher Alan; Mejia, Alfredo; Melgar-Quiñonez, Hugo; Balcazar, Hector G.; Anders, Robert L.; Segura-Pérez, Sofia; Duarte-Gardea, Maria O.; Ibarra, Jorge M.

    2010-01-01

    This cross-sectional study assessed: (a) awareness and knowledge of federal dietary tools (MyPyramid, Food Guide Pyramid (FGP), and food labels (FL)), and (b) the influence of acculturation and state of residence on FGP knowledge (FGPK) indicators among low-income Latina WIC participants (N = 479) living in Connecticut, Ohio, Texas, and California. Participants were familiar with FGP but only 37% recognized MyPyramid. FGPK was highest for fruits (71%) and lowest for the ‘breads and cereals’ group (12%). Less than half (47%) used FL’s when grocery shopping. Living in OH, was associated with the lowest FGPK. Multivariate analyses showed that more acculturated individuals living in CT/CA had better FGPK than participants living in TX and their less acculturated counterparts in CT/CA. The forthcoming revised federal dietary tools need to be adequately disseminated among Latinos, with special emphasis on those with lower acculturation levels, living in rapid emerging Latino communities or in the US–Mexico border. PMID:20549357

  7. Biomarkers of Safety and Immune Protection for Genetically Modified Live Attenuated Leishmania Vaccines Against Visceral Leishmaniasis – Discovery and Implications

    PubMed Central

    Gannavaram, Sreenivas; Dey, Ranadhir; Avishek, Kumar; Selvapandiyan, Angamuthu; Salotra, Poonam; Nakhasi, Hira L.

    2014-01-01

    Despite intense efforts there is no safe and efficacious vaccine against visceral leishmaniasis, which is fatal and endemic in many tropical countries. A major shortcoming in the vaccine development against blood-borne parasitic agents such as Leishmania is the inadequate predictive power of the early immune responses mounted in the host against the experimental vaccines. Often immune correlates derived from in-bred animal models do not yield immune markers of protection that can be readily extrapolated to humans. The limited efficacy of vaccines based on DNA, subunit, heat killed parasites has led to the realization that acquisition of durable immunity against the protozoan parasites requires a controlled infection with a live attenuated organism. Recent success of irradiated malaria parasites as a vaccine candidate further strengthens this approach to vaccination. We developed several gene deletion mutants in Leishmania donovani as potential live attenuated vaccines and reported extensively on the immunogenicity of LdCentrin1 deleted mutant in mice, hamsters, and dogs. Additional limited studies using genetically modified live attenuated Leishmania parasites as vaccine candidates have been reported. However, for the live attenuated parasite vaccines, the primary barrier against widespread use remains the absence of clear biomarkers associated with protection and safety. Recent studies in evaluation of vaccines, e.g., influenza and yellow fever vaccines, using systems biology tools demonstrated the power of such strategies in understanding the immunological mechanisms that underpin a protective phenotype. Applying similar tools in isolated human tissues such as PBMCs from healthy individuals infected with live attenuated parasites such as LdCen−/− in vitro followed by human microarray hybridization experiments will enable us to understand how early vaccine-induced gene expression profiles and the associated immune responses are coordinately regulated in normal individuals. In addition, comparative analysis of biomarkers in PBMCs from asymptomatic or healed visceral leishmaniasis individuals in response to vaccine candidates including live attenuated parasites may provide clues about determinants of protective immunity and be helpful in shaping the final Leishmania vaccine formulation in the clinical trials. PMID:24904589

  8. Biomarkers of safety and immune protection for genetically modified live attenuated leishmania vaccines against visceral leishmaniasis - discovery and implications.

    PubMed

    Gannavaram, Sreenivas; Dey, Ranadhir; Avishek, Kumar; Selvapandiyan, Angamuthu; Salotra, Poonam; Nakhasi, Hira L

    2014-01-01

    Despite intense efforts there is no safe and efficacious vaccine against visceral leishmaniasis, which is fatal and endemic in many tropical countries. A major shortcoming in the vaccine development against blood-borne parasitic agents such as Leishmania is the inadequate predictive power of the early immune responses mounted in the host against the experimental vaccines. Often immune correlates derived from in-bred animal models do not yield immune markers of protection that can be readily extrapolated to humans. The limited efficacy of vaccines based on DNA, subunit, heat killed parasites has led to the realization that acquisition of durable immunity against the protozoan parasites requires a controlled infection with a live attenuated organism. Recent success of irradiated malaria parasites as a vaccine candidate further strengthens this approach to vaccination. We developed several gene deletion mutants in Leishmania donovani as potential live attenuated vaccines and reported extensively on the immunogenicity of LdCentrin1 deleted mutant in mice, hamsters, and dogs. Additional limited studies using genetically modified live attenuated Leishmania parasites as vaccine candidates have been reported. However, for the live attenuated parasite vaccines, the primary barrier against widespread use remains the absence of clear biomarkers associated with protection and safety. Recent studies in evaluation of vaccines, e.g., influenza and yellow fever vaccines, using systems biology tools demonstrated the power of such strategies in understanding the immunological mechanisms that underpin a protective phenotype. Applying similar tools in isolated human tissues such as PBMCs from healthy individuals infected with live attenuated parasites such as LdCen(-/-) in vitro followed by human microarray hybridization experiments will enable us to understand how early vaccine-induced gene expression profiles and the associated immune responses are coordinately regulated in normal individuals. In addition, comparative analysis of biomarkers in PBMCs from asymptomatic or healed visceral leishmaniasis individuals in response to vaccine candidates including live attenuated parasites may provide clues about determinants of protective immunity and be helpful in shaping the final Leishmania vaccine formulation in the clinical trials.

  9. Genetically Modified Live Attenuated Leishmania donovani Parasites Induce Innate Immunity through Classical Activation of Macrophages That Direct the Th1 Response in Mice

    PubMed Central

    Bhattacharya, Parna; Dey, Ranadhir; Dagur, Pradeep K.; Kruhlak, Michael; Ismail, Nevien; Debrabant, Alain; Joshi, Amritanshu B.; Akue, Adovi; Kukuruga, Mark; Takeda, Kazuyo; Selvapandiyan, Angamuthu; McCoy, John Philip

    2015-01-01

    Visceral leishmaniasis (VL) causes significant mortality and there is no effective vaccine. Previously, we have shown that genetically modified Leishmania donovani parasites, here described as live attenuated parasites, induce a host protective adaptive immune response in various animal models. In this study, we demonstrate an innate immune response upon infection with live attenuated parasites in macrophages from BALB/c mice both in vitro and in vivo. In vitro infection of macrophages with live attenuated parasites (compared to that with wild-type [WT] L. donovani parasites) induced significantly higher production of proinflammatory cytokines (tumor necrosis factor alpha [TNF-α], interleukin-12 [IL-12], gamma interferon [IFN-γ], and IL-6), chemokines (monocyte chemoattractant protein 1/CCL-2, macrophage inflammatory protein 1α/CCL-3, and IP-10), reactive oxygen species (ROS), and nitric oxide, while concomitantly reducing anti-inflammatory cytokine IL-10 and arginase-1 activities, suggesting a dominant classically activated/M1 macrophage response. The classically activated response in turn helps in presenting antigen to T cells, as observed with robust CD4+ T cell activation in vitro. Similarly, parasitized splenic macrophages from live attenuated parasite-infected mice also demonstrated induction of an M1 macrophage phenotype, indicated by upregulation of IL-1β, TNF-α, IL-12, and inducible nitric oxide synthase 2 and downregulation of genes associated with the M2 phenotype, i.e., the IL-10, YM1, Arg-1, and MRC-1 genes, compared to WT L. donovani-infected mice. Furthermore, an ex vivo antigen presentation assay showed macrophages from live attenuated parasite-infected mice induced higher IFN-γ and IL-2 but significantly less IL-10 production by ovalbumin-specific CD4+ T cells, resulting in proliferation of Th1 cells. These data suggest that infection with live attenuated parasites promotes a state of classical activation (M1 dominant) in macrophages that leads to the generation of protective Th1 responses in BALB/c mice. PMID:26169275

  10. Genetically Modified Live Attenuated Leishmania donovani Parasites Induce Innate Immunity through Classical Activation of Macrophages That Direct the Th1 Response in Mice.

    PubMed

    Bhattacharya, Parna; Dey, Ranadhir; Dagur, Pradeep K; Kruhlak, Michael; Ismail, Nevien; Debrabant, Alain; Joshi, Amritanshu B; Akue, Adovi; Kukuruga, Mark; Takeda, Kazuyo; Selvapandiyan, Angamuthu; McCoy, John Philip; Nakhasi, Hira L

    2015-10-01

    Visceral leishmaniasis (VL) causes significant mortality and there is no effective vaccine. Previously, we have shown that genetically modified Leishmania donovani parasites, here described as live attenuated parasites, induce a host protective adaptive immune response in various animal models. In this study, we demonstrate an innate immune response upon infection with live attenuated parasites in macrophages from BALB/c mice both in vitro and in vivo. In vitro infection of macrophages with live attenuated parasites (compared to that with wild-type [WT] L. donovani parasites) induced significantly higher production of proinflammatory cytokines (tumor necrosis factor alpha [TNF-α], interleukin-12 [IL-12], gamma interferon [IFN-γ], and IL-6), chemokines (monocyte chemoattractant protein 1/CCL-2, macrophage inflammatory protein 1α/CCL-3, and IP-10), reactive oxygen species (ROS), and nitric oxide, while concomitantly reducing anti-inflammatory cytokine IL-10 and arginase-1 activities, suggesting a dominant classically activated/M1 macrophage response. The classically activated response in turn helps in presenting antigen to T cells, as observed with robust CD4(+) T cell activation in vitro. Similarly, parasitized splenic macrophages from live attenuated parasite-infected mice also demonstrated induction of an M1 macrophage phenotype, indicated by upregulation of IL-1β, TNF-α, IL-12, and inducible nitric oxide synthase 2 and downregulation of genes associated with the M2 phenotype, i.e., the IL-10, YM1, Arg-1, and MRC-1 genes, compared to WT L. donovani-infected mice. Furthermore, an ex vivo antigen presentation assay showed macrophages from live attenuated parasite-infected mice induced higher IFN-γ and IL-2 but significantly less IL-10 production by ovalbumin-specific CD4(+) T cells, resulting in proliferation of Th1 cells. These data suggest that infection with live attenuated parasites promotes a state of classical activation (M1 dominant) in macrophages that leads to the generation of protective Th1 responses in BALB/c mice. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  11. Deformed shell model calculations of half lives for β+/EC decay and 2ν β+β+/β+EC/ECEC decay in medium-heavy N~Z nuclei

    NASA Astrophysics Data System (ADS)

    Mishra, S.; Shukla, A.; Sahu, R.; Kota, V. K. B.

    2008-08-01

    The β+/EC half-lives of medium heavy N~Z nuclei with mass number A~64-80 are calculated within the deformed shell model (DSM) based on Hartree-Fock states by employing a modified Kuo interaction in (2p3/2,1f5/2,2p1/2,1g9/2) space. The DSM model has been quite successful in predicting many spectroscopic properties of N~Z medium heavy nuclei with A~64-80. The calculated β+/EC half-lives, for prolate and oblate shapes, compare well with the predictions of the calculations with Skyrme force by Sarriguren Going further, following recent searches, half-lives for 2ν β+β+/β+EC/ECEC decay for the nucleus Kr78 are calculated using DSM and the results compare well with QRPA predictions.

  12. Coherent control of an opsin in living brain tissue

    NASA Astrophysics Data System (ADS)

    Paul, Kush; Sengupta, Parijat; Ark, Eugene D.; Tu, Haohua; Zhao, Youbo; Boppart, Stephen A.

    2017-11-01

    Retinal-based opsins are light-sensitive proteins. The photoisomerization reaction of these proteins has been studied outside cellular environments using ultrashort tailored light pulses. However, how living cell functions can be modulated via opsins by modifying fundamental nonlinear optical properties of light interacting with the retinal chromophore has remained largely unexplored. We report the use of chirped ultrashort near-infrared pulses to modulate light-evoked ionic current from Channelrhodopsin-2 (ChR2) in brain tissue, and consequently the firing pattern of neurons, by manipulating the phase of the spectral components of the light. These results confirm that quantum coherence of the retinal-based protein system, even in a living neuron, can influence its current output, and open up the possibilities of using designer-tailored pulses for controlling molecular dynamics of opsins in living tissue to selectively enhance or suppress neuronal function for adaptive feedback-loop applications in the future.

  13. Interaction of carbohydrate modified boron nitride nanotubes with living cells.

    PubMed

    Emanet, Melis; Şen, Özlem; Çobandede, Zehra; Çulha, Mustafa

    2015-10-01

    Boron nitride nanotubes (BNNTs) are composed of boron and nitrogen atoms and they show significantly different properties from their carbon analogues (carbon nanotubes, CNTs). Due to their unique properties including low electrical conductivity, and imaging contrast and neutron capture properties; they can be used in biomedical applications. When their use in biological fields is considered, the route of their toxic effect should be clarified. Therefore, the study of interactions between BNNTs and living systems is important in envisaging biological applications at both cellular and sub-cellular levels to fully gain insights of their potential adverse effects. In this study, BNNTs were modified with lactose, glucose and starch and tested for their cytotoxicity. First, the interactions and the behavior of BNNTs with bovine serum albumin (BSA), Dulbecco's Modified Eagle's Medium (DMEM) and DMEM/Nutrient Mixture F-12Ham were investigated. Thereafter, their cellular uptake and the cyto- and genotoxicity on human dermal fibroblasts (HDFs) and adenocarcinoma human alveolar basal epithelial cells (A549) were evaluated. HDFs and A549 cells internalized the modified and unmodified BNNTs, and BNNTs were found to not cause significant viability change and DNA damage. A higher uptake rate of BNNTs by A549 cells compared to HDFs was observed. Moreover, a concentration-dependent cytotoxicity was observed on A549 cells while they were safer for HDFs in the same concentration range. Based on these findings, it can be concluded that BNNTs and their derivatives made with biomacromolecules might be good candidates for several applications in medicine and biomedical applications. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Core outcome measures for interventions to prevent or slow the progress of dementia for people living with mild to moderate dementia: Systematic review and consensus recommendations

    PubMed Central

    Groskreutz, Derek; Grinbergs-Saull, Anna; Howard, Rob; O’Brien, John T.; Mountain, Gail; Woods, Bob; Perneczky, Robert; McCleery, Jenny; Pickett, James; Challis, David; Charlesworth, Georgina; Featherstone, Katie; Jones, Roy; Schneider, Justine; Shepperd, Sasha; Thompson-Coon, Jo; Ballard, Clive; Burns, Alistair; Garrard, Peter; Kehoe, Patrick; Passmore, Peter; Robinson, Louise

    2017-01-01

    Background There are no disease-modifying treatments for dementia. There is also no consensus on disease modifying outcomes. We aimed to produce the first evidence-based consensus on core outcome measures for trials of disease modification in mild-to-moderate dementia. Methods and findings We defined disease-modification interventions as those aiming to change the underlying pathology. We systematically searched electronic databases and previous systematic reviews for published and ongoing trials of disease-modifying treatments in mild-to-moderate dementia. We included 149/22,918 of the references found; with 81 outcome measures from 125 trials. Trials involved participants with Alzheimer’s disease (AD) alone (n = 111), or AD and mild cognitive impairment (n = 8) and three vascular dementia. We divided outcomes by the domain measured (cognition, activities of daily living, biological markers, neuropsychiatric symptoms, quality of life, global). We calculated the number of trials and of participants using each outcome. We detailed psychometric properties of each outcome. We sought the views of people living with dementia and family carers in three cities through Alzheimer’s society focus groups. Attendees at a consensus conference (experts in dementia research, disease-modification and harmonisation measures) decided on the core set of outcomes using these results. Recommended core outcomes were cognition as the fundamental deficit in dementia and to indicate disease modification, serial structural MRIs. Cognition should be measured by Mini Mental State Examination or Alzheimer's Disease Assessment Scale-Cognitive Subscale. MRIs would be optional for patients. We also made recommendations for measuring important, but non-core domains which may not change despite disease modification. Limitations Most trials were about AD. Specific instruments may be superseded. We searched one database for psychometric properties. Interpretation This is the first review to identify the 81 outcome measures the research community uses for disease-modifying trials in mild-to-moderate dementia. Our recommendations will facilitate designing, comparing and meta-analysing disease modification trials in mild-to-moderate dementia, increasing their value. Trial registration PROSPERO no. CRD42015027346. PMID:28662127

  15. Genetically Modified Porcine Skin Grafts for Treatment of Severe Burn Injuries

    DTIC Science & Technology

    2011-07-01

    limited the usefulness of living porcine grafts, since the lack of blood supply soon lead to desiccation and avascular necrosis . Failure of...porcine grafts, since the lack of blood supply soon lead to desiccation and avascular necrosis . Failure of vascularization of xenografts is largely

  16. Composition of air pollution particles modifies oxidative stress in cells, tissues, and living systems

    EPA Science Inventory

    Epidemiological studies demonstrate an association between increased levels of ambient air pollution particles and human morbidity and mortality. Production of oxidants, either directly by the air pollution particles or by the host response to the particles, appears to be fundame...

  17. Investigating the Effects of Higher Spatial Resolution on Benthic Classification Accuracy at Midway Atoll

    DTIC Science & Technology

    2008-09-01

    2 X Components: 1 Y Components: 1 Product MBR Geographic Coordinates Number of Coordinates: 4 Coordinate: 1 Latitude...bottom (other than live coral) bldgs., docks, etc.) 4. linear reef- B. SHORELINE -INTERTIDAL modifiers 5. pinnacle reef- c. submerged vegetation- sand

  18. Total laparoscopic living donor right hepatectomy.

    PubMed

    Han, Ho-Seong; Cho, Jai Young; Yoon, Yoo-Seok; Hwang, Dae Wook; Kim, Young Ki; Shin, Hong Kyung; Lee, Woohyung

    2015-01-01

    Right lobe living donor liver transplantation (LDLT) is the predominant form of adult-to-adult LDLT. Accordingly, cosmetic and functional demand by young donors is increasing. We developed the world first total laparoscopic donor right hepatectomy (LDRH) in adult living donors. Total LDRH was performed in two young donors without vascular clamping. Modified extended right graft (right liver including all the middle hepatic vein branches) was retrieved from suprapubic transverse incision. After full mobilization of right liver, hilar dissection was done. First, right portal vein was isolated under retracting common bile duct laterally. Right hepatic artery was cautiously dissected and isolated without injuring. An exact transection line was drawn during transient clamping of the hepatic artery and portal vein on the right side of the liver using bulldog clamp. Dissection was meticulously performed along the right side of the middle hepatic vein until the origin of middle hepatic vein until exposure of the hilar plate. Anterior section vein branches (V5 and V8) were finely dissected and were reconstructed using an artificial vascular graft. A modified extended right graft with preservation of the middle hepatic vein branches was extracted through the suprapubic incision. There was no complication in both donors and recipients. Postoperative hospital stay of donors was 10 and 8 days, respectively. After follow-up of more than 1 year, all donors and recipients live well with normal liver function. Total LDRH was feasible in selected adult donors. If this procedure will be more standardized, then total LDRH will be new option for adult LDLT, which meets demand by donors and diminish guilty feeling by recipients.

  19. Endocannabinoids: Multi-scaled, Global Homeostatic Regulators of Cells and Society

    NASA Astrophysics Data System (ADS)

    Melamede, Robert

    Living systems are far from equilibrium open systems that exhibit many scales of emergent behavior. They may be abstractly viewed as a complex weave of dissipative structures that maintain organization by passing electrons from reduced hydrocarbons to oxygen. Free radicals are unavoidable byproducts of biological electron flow. Due to their highly reactive chemical properties, free radicals modify all classes of biological molecules (carbohydrates, lipids, nucleic acids, and proteins). As a result, free radicals are destructive. The generally disruptive nature of free radicals makes them the "friction of life." As such, they are believed to be the etiological agents behind age related illnesses such as cardiovascular, immunological, and neurological diseases, cancer, and ageing itself. Free radicals also play a critical constructive role in living systems. From a thermodynamic perspective, life can only exist if a living system takes in sufficient negative entropy from its environment to overcome the obligatory increase in entropy that would result if the system could not appropriately exchange mass, energy and information with its environment. Free radicals are generated in response to perturbations in the relationship between a living system and its environment. However, evolution has selected for biological response systems to free radicals so that the cellular biochemistry can adapt to environmental perturbations by modifying cellular gene expression and biochemistry. Endocannabinoids are marijuana-like compounds that have their origins hundreds of millions of years in the evolutionary past. They serve as fundamental modulators of energy homeostasis in all vertebrates. Their widespread biological activities may often be attributed to their ability to minimize the negative consequences of free radicals.

  20. Modified constraint-induced therapy for children with hemiplegic cerebral palsy: a randomized trial.

    PubMed

    Wallen, Margaret; Ziviani, Jenny; Naylor, Olivia; Evans, Ruth; Novak, Iona; Herbert, Robert D

    2011-12-01

    Conventional constraint-based therapies are intensive and demanding to implement, particularly for children. Modified forms of constraint-based therapies that are family-centred may be more acceptable and feasible for families of children with cerebral palsy (CP)-but require rigorous evaluation using randomized trials. The aim of this study was to determine the effects of modified constraint-induced therapy compared with intensive occupational therapy on activities of daily living and upper limb outcomes in children with hemiplegic CP. In this assessor-blinded pragmatic randomized trial, 50 children (27 males, 23 females; age range 19 mo-7 y 10 mo) with hemiplegic CP were randomized using a concealed allocation procedure to one of two 8-week interventions: intensive occupational therapy (n = 25), or modified constraint-induced therapy (n = 25). Manual Ability Classification System (MACS) levels of the participants were, level I n = 2, II n = 37, III n = 8, and level IV n = 1; Gross Motor Function Classification System (GMFCS) levels were, level I n = 33, level II n = 15, and level III n = 1. Participants were recruited through three specialist CP centres in Australia and randomized between January 2008 and April 2010. Children randomized to modified constraint-induced therapy wore a mitt on the unaffected hand for 2 hours each day, during which time the children participated in targeted therapy. The primary outcome was the Canadian Occupational Performance Measure (COPM--measured on a 10-point scale) at completion of therapy. Other outcome measures were Goal Attainment Scaling, Assisting Hand Assessment, Pediatric Motor Activity Log, Modified Ashworth Scale, Modified Tardieu Scale, and a parent questionnaire. Assessments were carried out at 10 weeks and 6 months following randomization. All participants were included in the analysis. Between-group differences for all outcomes were neither clinically important nor statistically significant. The mean difference in COPM was 0.3 (95% confidence interval [CI] -0.8 to 1.4; p=0.61) and mean difference in COPM satisfaction was 0.1 (95% CI -1.1 to 1.2; p=0.90). Minor adverse events were reported by five of the 25 participants in the modified constraint-induced therapy group and by one of the 25 in the intensive occupational therapy group. All adverse events were related to participants' lack of acceptance of therapy. Modified constraint-induced therapy is no more effective than intensive occupational therapy for improving completion of activities of daily living or upper limb function in children with hemiplegic CP. © The Authors. Developmental Medicine & Child Neurology © 2011 Mac Keith Press.

  1. Use of Traditional and Genetically Modified Probiotics in Human Health: What Does the Future Hold?

    PubMed

    Bermúdez-Humarán, Luis G; Langella, Philippe

    2017-09-01

    Probiotics are live, nonpathogenic microorganisms that confer benefits to human health when administered in adequate amounts. Among the frequent proposed health benefits attributed to probiotics, their ability to interact with the host immune system is now well demonstrated. Although history has revealed that probiotics were part of fermented foods in the past, clinicians have started to use them therapeutically in regular diets. Moreover, the use of genetically modified probiotics to deliver molecules of therapeutic interest is gaining importance as an extension of the probiotic concept. This chapter summarizes some of the recent findings and perspectives on the use of both traditional and genetically modified probiotics to treat human diseases as well as what the future may hold concerning the use of these probiotics in humans.

  2. Photoluminescent graphene quantum dots for in vivo imaging of apoptotic cells

    NASA Astrophysics Data System (ADS)

    Roy, Prathik; Periasamy, Arun Prakash; Lin, Chiu-Ya; Her, Guor-Mour; Chiu, Wei-Jane; Li, Chi-Lin; Shu, Chia-Lun; Huang, Chih-Ching; Liang, Chi-Te; Chang, Huan-Tsung

    2015-01-01

    Apoptosis (programmed cell death) is linked to many incurable neurodegenerative, cardiovascular and cancer causing diseases. Numerous methods have been developed for imaging apoptotic cells in vitro; however, there are few methods available for imaging apoptotic cells in live animals (in vivo). Here we report a novel method utilizing the unique photoluminescence properties of plant leaf-derived graphene quantum dots (GQDs) modified with annexin V antibody (AbA5) to form (AbA5)-modified GQDs (AbA5-GQDs) enabling us to label apoptotic cells in live zebrafish (Danio rerio). The key is that zebrafish shows bright red photoluminescence in the presence of apoptotic cells. The toxicity of the GQDs has also been investigated with the GQDs exhibiting high biocompatibility as they were excreted from the zebrafish's body without affecting its growth significantly at a concentration lower than 2 mg mL-1 over a period of 4 to 72 hour post fertilization. The GQDs have further been used to image human breast adenocarcinoma cell line (MCF-7 cells), human cervical cancer cell line (HeLa cells), and normal human mammary epithelial cell line (MCF-10A). These results are indispensable to further the advance of graphene-based nanomaterials for biomedical applications.Apoptosis (programmed cell death) is linked to many incurable neurodegenerative, cardiovascular and cancer causing diseases. Numerous methods have been developed for imaging apoptotic cells in vitro; however, there are few methods available for imaging apoptotic cells in live animals (in vivo). Here we report a novel method utilizing the unique photoluminescence properties of plant leaf-derived graphene quantum dots (GQDs) modified with annexin V antibody (AbA5) to form (AbA5)-modified GQDs (AbA5-GQDs) enabling us to label apoptotic cells in live zebrafish (Danio rerio). The key is that zebrafish shows bright red photoluminescence in the presence of apoptotic cells. The toxicity of the GQDs has also been investigated with the GQDs exhibiting high biocompatibility as they were excreted from the zebrafish's body without affecting its growth significantly at a concentration lower than 2 mg mL-1 over a period of 4 to 72 hour post fertilization. The GQDs have further been used to image human breast adenocarcinoma cell line (MCF-7 cells), human cervical cancer cell line (HeLa cells), and normal human mammary epithelial cell line (MCF-10A). These results are indispensable to further the advance of graphene-based nanomaterials for biomedical applications. Electronic supplementary information (ESI) available: Experimental discussion on synthesis, characterization, cellular imaging, cytotoxicity of GQDs in addition to its effect on zebrafish embryos, preparation of annexin V (A5)-modified GQDs (AbA5-GQDs), staining procedures and imaging are given. Figures for XRD, UV-vis absorption, photoluminescence of GQDs, mortality of zebrafish, time course recording of morphology of zebrafish embryos and morphology of adult zebrafish exposed to GQDs are illustrated. See DOI: 10.1039/c4nr07005d

  3. 50 CFR 622.48 - Adjustment of management measures.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... of the applicable FMPs, the RA may establish or modify the following items: (a) Caribbean coral reef... (maintaining fish in whole condition, use as bait). (k) South Atlantic coral, coral reefs, and live/hard bottom... conservation districts. (b) Caribbean reef fish. Fishery management units (FMUs), quotas, trip limits, bag...

  4. EFFECT OF ACUTE STRESS ON PLASMA CONCENTRATIONS OF SEX AND STRESS HORMONES IN JUVENILE ALLIGATORS LIVING IN CONTROL AND CONTAMINATED LAKES

    EPA Science Inventory

    Environmental contaminants can act as stressors, inducing elevated circulating concentrations of stress hormones such as corticosterone and cortisol. Development in contaminated eggs has been reported to modify circulating sex steroid hormone concentrations in alligators (Alligat...

  5. 50 CFR 622.48 - Adjustment of management measures.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... of the applicable FMPs, the RA may establish or modify the following items: (a) Caribbean coral reef..., and essential fish habitat HAPCs or Coral HAPCs. (d) Gulf reef fish. (1) For a species or species..., coral reefs, and live/hard bottom habitats. Definitions of essential fish habitat and essential fish...

  6. Along Came a Spider: Using Live Arthropods in a Predator-Prey Activity

    ERIC Educational Resources Information Center

    Richardson, Matthew L.; Hari, Janice

    2011-01-01

    We developed a predator-prey activity with eighth-grade students in which they used wolf spiders ("Lycosa carolinensis"), house crickets ("Acheta domestica"), and abiotic factors to address how (1) adaptations in predators and prey shape their interaction and (2) abiotic factors modify the interaction between predators and…

  7. AROMATASE ACTIVITY IN THE OVARY OF MOSQUITOFISH GAMBUSIA HOLBROOKI, COLLECTED FROM THE FENHOLLOWAY AND ECONFINA RIVERS, FLORIDA (

    EPA Science Inventory

    Scientists are increasingly aware of the adverse effects of environmental contaminants, including their ability to alter the normal development and reproduction of wildlife species by modifying the endocrine system. Female mosquitofish living downstream of a paper mill plant loca...

  8. Nasal lavage natural killer cell function is suppressed in smokers after live attenuated influenza virus

    EPA Science Inventory

    Background Modified function of immune cells in nasal secretions may playa role in the enhanced susceptibility to resp iratory viruses that is seen in smokers. Innate immune cells in nasal secretions have largely been characterized by cellular differentials using morphologic c...

  9. Sequence Stability of PRRSV Chimeras After Passage in Swine

    USDA-ARS?s Scientific Manuscript database

    Recombinant chimeric porcine reproductive and respiratory syndrome virus (PRRSV), generated from parental strains MN184 and a licensed modified live vaccine (Ingelvac® PRRS MLV), and a MN184 nsp2 deletion mutant were used to elucidate the mechanisms of attenuation and/or protective immunity to heter...

  10. Modifying the Existing Campus Building for Accessibility: Accessible Products Catalog.

    ERIC Educational Resources Information Center

    Cotler, Stephen Richard

    This catalog is intended to assist architects and college administrators to select products that help physically handicapped people lead lives free of architectural barriers. The product information, obtained directly from the manufacturers, is listed on comparative matrix sheets, that can be used to achieve the design recommendations. Products of…

  11. Alternative strategies for the control and elimination of PRRS

    USDA-ARS?s Scientific Manuscript database

    Porcine reproductive and respiratory syndrome (PRRS) is the most costly disease of modern global pig production systems. The etiological agent, PRRS virus (PRRSV), an RNA virus, was identified in Europe (PRRSV-1 isolates) in 1991, and later in the US (PRRSV-2 isolates). Modified live virus (MLV) vac...

  12. Modified live Edwardsiella ictaluri vaccine, AQUAVAC-ESC, lacks multidrug resistance plasmids

    USDA-ARS?s Scientific Manuscript database

    Plasmid mediated antibiotic resistance was first discovered in Edwardsiella ictaluri in the early 1990’s, and in 2007 an E. ictaluri isolate harboring an IncA/C plasmid was recovered from a moribund channel catfish infected with the bacterium. Due to the identification of multidrug resistance plasm...

  13. Development and Validation of an Instrument to Measure University Students' Biotechnology Attitude

    ERIC Educational Resources Information Center

    Erdogan, Mehmet; Ozel, Murat; Usak, Muhammet; Prokop, Pavol

    2009-01-01

    The impact of biotechnologies on peoples' everyday lives continuously increases. Measuring young peoples' attitudes toward biotechnologies is therefore very important and its results are useful not only for science curriculum developers and policy makers, but also for producers and distributors of genetically modified products. Despite of…

  14. Undergraduate Female Science-Related Career Choices: A Phenomenological Study

    ERIC Educational Resources Information Center

    Curry, Kathy S.

    2010-01-01

    This qualitative phenomenological study used a modified Groenewald's five steps method with semi-structured, recorded, and transcribed interviews to focus on the underrepresentation of females in science-related careers. The study explored the lived experiences of a purposive sample of 25 senior female college students attending a college in…

  15. Vaccines and Disease-Modifying Antirheumatic Drugs: Practical Implications for the Rheumatologist.

    PubMed

    Friedman, Marcia A; Winthrop, Kevin L

    2017-02-01

    Patients with rheumatoid arthritis are highly vulnerable to infections because of abnormalities in their immune system, and because of immunosuppressive effects of their medications. Vaccinations in this population are complicated by disease-modifying antirheumatic drugs, which also modulate or suppress the immune system and potentially decrease the immunogenicity and efficacy of the vaccines. We review the available data regarding the impact of rheumatoid arthritis therapy on the immunogenicity of various common vaccines. We also review rheumatoid arthritis-specific vaccination recommendations, live vaccine safety concerns, and current gaps in our understanding of these issues." Published by Elsevier Inc.

  16. Nucleoside-modified mRNA vaccines induce potent T follicular helper and germinal center B cell responses.

    PubMed

    Pardi, Norbert; Hogan, Michael J; Naradikian, Martin S; Parkhouse, Kaela; Cain, Derek W; Jones, Letitia; Moody, M Anthony; Verkerke, Hans P; Myles, Arpita; Willis, Elinor; LaBranche, Celia C; Montefiori, David C; Lobby, Jenna L; Saunders, Kevin O; Liao, Hua-Xin; Korber, Bette T; Sutherland, Laura L; Scearce, Richard M; Hraber, Peter T; Tombácz, István; Muramatsu, Hiromi; Ni, Houping; Balikov, Daniel A; Li, Charles; Mui, Barbara L; Tam, Ying K; Krammer, Florian; Karikó, Katalin; Polacino, Patricia; Eisenlohr, Laurence C; Madden, Thomas D; Hope, Michael J; Lewis, Mark G; Lee, Kelly K; Hu, Shiu-Lok; Hensley, Scott E; Cancro, Michael P; Haynes, Barton F; Weissman, Drew

    2018-06-04

    T follicular helper (Tfh) cells are required to develop germinal center (GC) responses and drive immunoglobulin class switch, affinity maturation, and long-term B cell memory. In this study, we characterize a recently developed vaccine platform, nucleoside-modified, purified mRNA encapsulated in lipid nanoparticles (mRNA-LNPs), that induces high levels of Tfh and GC B cells. Intradermal vaccination with nucleoside-modified mRNA-LNPs encoding various viral surface antigens elicited polyfunctional, antigen-specific, CD4 + T cell responses and potent neutralizing antibody responses in mice and nonhuman primates. Importantly, the strong antigen-specific Tfh cell response and high numbers of GC B cells and plasma cells were associated with long-lived and high-affinity neutralizing antibodies and durable protection. Comparative studies demonstrated that nucleoside-modified mRNA-LNP vaccines outperformed adjuvanted protein and inactivated virus vaccines and pathogen infection. The incorporation of noninflammatory, modified nucleosides in the mRNA is required for the production of large amounts of antigen and for robust immune responses. © 2018 Pardi et al.

  17. Preparation and properties of thermal insulation coatings with a sodium stearate-modified shell powder as a filler

    NASA Astrophysics Data System (ADS)

    Tang, Qiang; Zhang, Ya-mei; Zhang, Pei-gen; Shi, Jin-jie; Tian, Wu-bian; Sun, Zheng-ming

    2017-10-01

    Waste shell stacking with odor and toxicity is a serious hazard to our living environment. To make effective use of the natural resources, the shell powder was applied as a filler of outdoor thermal insulation coatings. Sodium stearate (SS) was used to modify the properties of shell powder to reduce its agglomeration and to increase its compatibility with the emulsion. The oil absorption rate and the spectrum reflectance of the shell powder show that the optimized content of SS as a modifier is 1.5wt%. The total spectrum reflectance of the coating made with the shell powder that is modified at this optimum SS content is 9.33% higher than that without any modification. At the optimum SS content of 1.5wt%, the thermal insulation of the coatings is improved by 1.0°C for the cement mortar board and 1.6°C for the steel plate, respectively. The scouring resistance of the coating with the 1.5wt% SS-modified shell powder is three times that of the coating without modification.

  18. Evaluation of Psn, HmuR and a modified LcrV protein delivered to mice by live attenuated Salmonella as a vaccine against bubonic and pneumonic Yersinia pestis challenge.

    PubMed

    Branger, Christine G; Sun, Wei; Torres-Escobar, Ascención; Perry, Robert; Roland, Kenneth L; Fetherston, Jacqueline; Curtiss, Roy

    2010-12-16

    We evaluated the ability of Yersinia pestis antigens HmuR, Psn and modified forms of LcrV delivered by live attenuated Salmonella strains to stimulate a protective immune response against subcutaneous or intranasal challenge with Y. pestis CO92. LcrV196 is a previously described truncated protein that includes aa 131-326 of LcrV and LcrV5214 has been modified to replace five key amino acids required for interaction with the TLR2 receptor. Psn is the outer membrane receptor for the siderophore, yersiniabactin, and the bacteriocin, pesticin. Mice immunized with Salmonella synthesizing Psn, LcrV196 or LcrV5214 developed serum IgG responses to the respective Yersinia antigen and were protected against pneumonic challenge with Y. pestis. Immunization with Salmonella synthesizing Psn or LcrV196 was sufficient to afford nearly full protection against bubonic challenge, while immunization with the strain synthesizing LcrV5214 was not protective. Immunization with Salmonella synthesizing HmuR, an outer membrane protein involved in heme acquisition in Y. pestis, was poorly immunogenic and did not elicit a protective response against either challenge route. These findings indicate that both Psn and LcrV196 delivered by Salmonella provide protection against both bubonic and pneumonic plague. Copyright © 2010 Elsevier Ltd. All rights reserved.

  19. Safe sorting of GFP-transduced live cells for subsequent culture using a modified FACS vantage.

    PubMed

    Sørensen, T U; Gram, G J; Nielsen, S D; Hansen, J E

    1999-12-01

    A stream-in-air cell sorter enables rapid sorting to a high purity, but it is not well suited for sorting of infectious material due to the risk of airborne spread to the surroundings. A FACS Vantage cell sorter was modified for safe use with potentially HIV infected cells. Safety tests with bacteriophages were performed to evaluate the potential spread of biologically active material during cell sorting. Cells transduced with a retroviral vector carrying the gene for GFP were sorted on the basis of their GFP fluorescence, and GFP expression was followed during subsequent culture. The bacteriophage sorting showed that the biologically active material was confined to the sorting chamber. A failure mode simulating a nozzle blockage resulted in detectable droplets inside the sorting chamber, but no droplets could be detected when an additional air suction from the sorting chamber had been put on. The GFP transduced cells were sorted to 99% purity. Cells not expressing GFP at the time of sorting did not turn on the gene during subsequent culture. Un-sorted cells and cells sorted to be positive for GFP showed a decrease in the fraction of GFP positive cells during culture. Sorting of live infected cells can be performed safely and with no deleterious effects on vector expression using the modified FACS Vantage instrument. Copyright 1999 Wiley-Liss, Inc.

  20. SCALE-UP Your Astronomy and Physics Undergraduate Courses to Incorporate Heliophysics

    NASA Astrophysics Data System (ADS)

    Al-Rawi, Ahlam N.; Cox, Amanda; Hoshino, Laura; Fitzgerald, Cullen; Cebulka, Rebecca; Rodriguez Garrigues, Alvar; Montgomery, Michele; Velissaris, Chris; Flitsiyan, Elena

    2016-01-01

    Although physics and astronomy courses include heliophysics topics, students still leave these courses without knowing what heliophysics is and how heliophysics relates to their daily lives. To meet goals of NASA's Living With a Star Program of incorporating heliophysics into undergraduate curriculum, UCF Physics has modified courses such as Astronomy (for non-science majors), Astrophysics, and SCALE-UP: Electricity and Magnetism for Engineers and Scientists to incorporate heliophysics topics. In this presentation, we discuss these incorporations and give examples that have been published in NASA Wavelength. In an associated poster, we present data on student learnin

  1. The novel use of modified pig zygotic medium for the efficient culture of the preimplantation mouse embryos.

    PubMed

    Amarnath, Dasari; Wakayama, Sayaka; Zhu, Jie; Moawad, Adel R; Wakayama, Teruhiko; Campbell, Keith H S

    2011-12-01

    A high potassium concentration in culture media is considered detrimental to in vitro culture of mouse embryos. Here we show that pig zygotic medium (PZM) containing a higher concentration of potassium, and modified to contain 0.2 mM glucose and 0.01 mM EDTA, supported efficient pre- and post-implantation development of mouse zygotes to blastocysts and live pups, respectively. At first, modified PZM (mPZM) was compared with other culture media such as M16, CZB and KSOM-AA for its ability to support development of in vivo mouse zygotes to the blastocyst stage. The proportions of zygotes reaching 2-cell (94-99%) and blastocyst (90-96%) stages in mPZM and other media were not different. However, hatching rates of blastocysts were different (P < 0.05); whereas more than 90% of the blastocysts were hatching in mPZM or KSOM-AA, only 60% of the blastocysts did in M16 or CZB media (P < 0.05). Next we compared post-implantation development of in vitro fertilized zygotes developed to blastocysts in mPZM and KSOM-AA. The proportion of blastocysts developing into live pups was not different between mPZM (49%) and KSOM-AA (44%). Finally, we evaluated whether mPZM could be also used as a fertilization medium. Modified PZM containing 5.56 mM of glucose and 0.4% BSA efficiently supported IVF of mouse gametes. The percent of zygotes cleaving to 2-cell (94-98%) and blastocysts (91-93%) stage was not different from zygotes fertilized in human tubal fluid medium. We concluded that modified pig zygotic medium containing a higher potassium concentration than any other commonly used mouse media supported not only culture of mouse embryos, but also efficient IVF of mouse gametes. Copyright © 2011 Elsevier Inc. All rights reserved.

  2. Implications for Social Support on Prolonged Sleep Difficulties among a Disaster-Affected Population: Second Report from a Cross-Sectional Survey in Ishinomaki, Japan

    PubMed Central

    Matsumoto, Shoko; Yamaoka, Kazue; Inoue, Machiko; Inoue, Mariko; Muto, Shinsuke

    2015-01-01

    Study Objectives This study aimed to investigate the role of social factors, especially social support for sleep, among victims living at home around 1–2 years after the Great East Japan Earthquake and tsunami. Design A cross-sectional household survey was conducted between May and December 2012 (14–21 months after the disaster) in the Ishinomaki area, Japan. Univariate and multivariate logistic regression models were used to examine the association between social factors, including social support, and prolonged sleep difficulties (persisting over 1 month). Social support was divided into three functions: emotional, informational, and instrumental support. Participants Data were obtained on 2,593 individuals who were living at home after the disaster. Results The prevalence of prolonged sleep difficulties was 6.9% (5.8% male, 7.7% female). This study showed that lack of social support has a stronger association with prolonged sleep difficulties than non-modifiable or hardly modifiable consequences caused directly by the disaster, i.e., severity of home damage, change in family structure and income. Among the three dimensions of social support, lack of emotional support showed the strongest association with prolonged sleep difficulties. Conclusions Social support, especially emotional support, may positively affect sleep among victims living at home around 1–2 years after a disaster. PMID:26087305

  3. Fluorescence Cross-Correlation Spectroscopy Reveals Mechanistic Insights into the Effect of 2′-O-Methyl Modified siRNAs in Living Cells

    PubMed Central

    Ohrt, Thomas; Staroske, Wolfgang; Mütze, Jörg; Crell, Karin; Landthaler, Markus; Schwille, Petra

    2011-01-01

    RNA interference (RNAi) offers a powerful tool to specifically direct the degradation of complementary RNAs, and thus has great therapeutic potential for targeting diseases. Despite the reported preferences of RNAi, there is still a need for new techniques that will allow for a detailed mechanistic characterization of RNA-induced silencing complex (RISC) assembly and activity to further improve the biocompatibility of modified siRNAs. In contrast to previous reports, we investigated the effects of 2′-O-methyl (2′OMe) modifications introduced at specific positions within the siRNA at the early and late stages of RISC assembly, as well as their influence on target recognition and cleavage directly in living cells. We found that six to 10 2′OMe nucleotides on the 3′-end inhibit passenger-strand release as well as target-RNA cleavage without changing the affinity, strand asymmetry, or target recognition. 2′OMe modifications introduced at the 5′-end reduced activated RISC stability, whereas incorporations at the cleavage site showed only minor effects on passenger-strand release when present on the passenger strand. Our new fluorescence cross-correlation spectroscopy assays resolve different steps and stages of RISC assembly and target recognition with heretofore unresolved detail in living cells, which is needed to develop therapeutic siRNAs with optimized in vivo properties. PMID:21689532

  4. Serologic evaluation, efficacy, and safety of a commerical modified-live canine distemper vaccine in domestic ferrets.

    PubMed

    Wimsatt, J; Jay, M T; Innes, K E; Jessen, M; Collins, J K

    2001-05-01

    To determine efficacy and safety of a commercial modified-live canine distemper virus (CDV) vaccine used for prophylaxis in domestic ferrets. Sixteen 16-week-old neutered male ferrets. Equal groups of ferrets were inoculated subcutaneously at 16 and 20 weeks of age with saline (0.9% NaCl) solution or a vaccine derived from the Onderstepoort CDV strain and attenuated in a primate cell line. Live virulent CDV was administered to all ferrets intranasally and orally 3 weeks after the second inoculation. Clinical signs and body weights were monitored regularly during the study. Blood samples for serologic examination were drawn prior to each inoculation, before challenge exposure, and 10, 15, and 21 days after exposure. Blood samples for reverse transcriptase polymerase chain reaction (RT-PCR) were obtained 5 days after the first vaccination, and 5, 10, 15, and 21 days after challenge exposure. After challenge exposure, control ferrets had significantly more clinical signs and weight loss, compared with vaccinates. All vaccinated ferrets survived, whereas all control ferrets died. The RT-PCR assay was successful in detecting CDV in blood and fresh or formalin-fixed tissues from infected ferrets. Findings suggest that the vaccine when given SC to domestic ferrets as directed is safe and protective against challenge exposure with virulent CDV. The RT-PCR assay may simplify detection of CDV in fresh and fixed tissues.

  5. Comparison between the resistance of benzalkonium chloride-adapted and -nonadapted biofilms of Listeria monocyogenes to modified atmosphere packaging and nisin once transferred to mussels.

    PubMed

    Saá Ibusquiza, P; Herrera, J J R; Cabo, M L

    2011-07-01

    Benzalkonium chloride-adapted and -nonadapted Listeria monocytogenes biofilm cells were transferred by contact to cooked or live mussels and packed in rich CO(2) and O(2), respectively. The viabilities of transferred cells during storage of these packed samples at 2.5 °C were compared. In addition, in cooked mussels the combined effect of CO(2) and nisin against the survival of L. monocytogenes was also studied by using a first-order factorial design. The results obtained demonstrated that biofilms formed by benzalkonium chloride-adapted L. monocytogenes cells could be more resistant to the application of modified atmospheres rich in CO(2) and nisin once they have been transferred to cooked mussels by contact (simulating cross-contamination). This implies an increase in the risk associated with the presence of these cells in food processing plants. Significant empirical equations obtained after 7, 11, and 20 days showed an inhibitory effect of CO(2) and nisin against L. monocytogenes. However, a significant positive interaction between both variables highlights an incompatibility between CO(2) and nisin at high concentrations. Results also demonstrated that L. monocytogenes could persist after cross-contamination during the processing of live mussels, so L. monocytogenes is of concern as a contaminant in live mussels packaged in high-O(2) atmospheres. Copyright ©, International Association for Food Protection

  6. Living Alone or With Others and Depressive Symptoms, and Effect Modification by Residential Social Cohesion Among Older Adults in Japan: The JAGES Longitudinal Study.

    PubMed

    Honjo, Kaori; Tani, Yukako; Saito, Masashige; Sasaki, Yuri; Kondo, Katsunori; Kawachi, Ichiro; Kondo, Naoki

    2018-02-03

    There is little longitudinal evidence on the impact of specific living arrangements (ie, who individuals live with) on mental health among older adults, and no studies have examined the modifying effect of residential social cohesion level on this association. We aimed to examine the association between living arrangements and depressive symptoms and whether this association varies with residential neighborhood social cohesion level among 19,656 men and 22,513 women aged 65 years and older in Japan. We analyzed the association between baseline living arrangements in 2010 and depressive symptoms in 2013. We calculated gender-specific odds ratios (ORs) of living arrangements for depressive symptoms using a logistic regression and conducted subgroup analyses by neighborhood social cohesion level. Among men (but not women), living alone (OR 1.43; 95% confidence intervals [CI], 1.18-1.73) and living with spouse and parent (OR 1.47, 95% CI, 1.09-1.98) were associated with increased odds of depressive symptoms compared with living with a spouse only. Living with spouse and child was a risk for men in the young age group but a protective factor for women. We also identified that the negative impact of living arrangements on depressive symptoms was attenuated in neighborhoods with higher levels of social cohesion. Living arrangements are associated with risk of depressive symptoms among men and women; these associations differ by gender and neighborhood social cohesion level. Our results suggest the need to pay more attention to whether individuals live alone, as well as who individuals live with, to prevent depressive symptoms among older adults.

  7. A comparative study of female sterilization via modified Uchida and silver clip techniques in rural China.

    PubMed

    Qiu, Hongyan; Li, Li; Wu, Shangchun; Liang, Hong; Yuan, Wei; He, Yingqin

    2011-03-01

    To compare the specific effects of 2 female sterilization methods: the modified Uchida technique and the application of silver clips. A total of 2198 women living in rural areas who were still of reproductive age but opting for sterilization were enrolled. The participants were randomly divided into 2 groups, and underwent sterilization by either modified Uchida technique or silver clips. Information on acceptability, operation conditions, effectiveness, adverse effects, and complaints was collected 3, 6, and 12 months after the procedure. No significant difference in effectiveness, adverse effects or chief complaints between the 2 procedures was found. Differences in operative outcome, bleeding volume during the procedure, and operation time were found. A shorter operation time and less bleeding for the silver clip method indicated that female sterilization by this technique was as safe as that by modified Uchida technique. Copyright © 2010 International Federation of Gynecology and Obstetrics. Published by Elsevier Ireland Ltd. All rights reserved.

  8. A study of the diffusion dynamics and concentration distribution of gold nanospheres (GNSs) without fluorescent labeling inside live cells using fluorescence single particle spectroscopy.

    PubMed

    Liu, Fangchao; Dong, Chaoqing; Ren, Jicun

    2018-03-15

    Colloidal gold nanospheres (GNSs) have become important nanomaterials in biomedical applications due to their special optical properties, good chemical stability, and biocompatibility. However, measuring the diffusion coefficients or concentration distribution of GNSs within live cells accurately without any extra fluorescent labeling in situ has still not been resolved. In this work, a single particle method is developed to study the concentration distribution of folic acid-modified GNSs (FA-GNSs) internalized via folate receptors, and investigates their diffusion dynamics within live cells using single particle fluorescence correlation spectroscopy (FCS). We optimized the experimental conditions and verified the feasibility of 30 nm GNSs without extra fluorescence labeling being used for single particle detection inside live cells. Then, the FCS characterization strategy was used to measure the concentration and diffusion coefficient distributions of GNSs inside live cells and the obtained results were basically in agreement with those obtained by TEM. The results demonstrate that our strategy is characterized as an in situ, nondestructive, rapid and dynamic method for the assay of live cells, and it may be widely used in the further design of GNP-based drug delivery and therapeutics.

  9. Event-based analysis of free-living behaviour.

    PubMed

    Granat, Malcolm H

    2012-11-01

    The quantification of free-living physical activities is important in understanding how physical activity and sedentary behaviour impact on health and also on how interventions might modify free-living behaviour to enhance health. Quantification, and the terminology used, has in many ways been determined by the choice of measurement technique. The inter-related issues around measurement devices and terminology used are explored. This paper proposes a terminology and a systematic approach for the analysis of free-living activity information using event-based activity data. The event-based approach uses a flexible hierarchical classification of events and, dependent on the research question, analysis can then be undertaken on a selection of these events. The quantification of free-living behaviour is therefore the result of the analysis on the patterns of these chosen events. The application of this approach is illustrated with results from a range of published studies by our group showing how event-based analysis provides a flexible yet robust method of addressing the research question(s) and provides a deeper insight into free-living behaviour. It is proposed that it is through event-based analysis we can more clearly understand how behaviour is related to health and also how we can produce more relevant outcome measures.

  10. Additive Manufacturing of Catalytically Active Living Materials.

    PubMed

    Saha, Abhijit; Johnston, Trevor G; Shafranek, Ryan T; Goodman, Cassandra J; Zalatan, Jesse G; Storti, Duane W; Ganter, Mark A; Nelson, Alshakim

    2018-04-25

    Living materials, which are composites of living cells residing in a polymeric matrix, are designed to utilize the innate functionalities of the cells to address a broad range of applications such as fermentation and biosensing. Herein, we demonstrate the additive manufacturing of catalytically active living materials (AMCALM) for continuous fermentation. A multi-stimuli-responsive yeast-laden hydrogel ink, based on F127-dimethacrylate, was developed and printed using a direct-write 3D printer. The reversible stimuli-responsive behaviors of the polymer hydrogel inks to temperature and pressure are critical, as they enabled the facile incorporation of yeast cells and subsequent fabrication of 3D lattice constructs. Subsequent photo-cross-linking of the printed polymer hydrogel afforded a robust elastic material. These yeast-laden living materials were metabolically active in the fermentation of glucose into ethanol for 2 weeks in a continuous batch process without significant reduction in efficiency (∼90% yield of ethanol). This cell immobilization platform may potentially be applicable toward other genetically modified yeast strains to produce other high-value chemicals in a continuous biofermentation process.

  11. Biosafety evaluation of recombinant live oral bacterial vaccines in the context of European regulation.

    PubMed

    Favre, Didier; Viret, Jean-François

    2006-05-01

    Live bacterial vaccines represent a highly valid preventive strategy in the fight against infectious disease. However, the road from research to market is peppered with hurdles, one of which is the requirement for high biosafety characteristics, which the candidate vaccine has to display. In Europe, the European Agency for the evaluation of medicinal products (EMEA) is the relevant authority regulating the licensure of genetically engineered vaccines. For this purpose, the agency may rely on several directives and guidelines defined in the past 15 years. As for live vaccines containing genetically modified organisms (GMOs) susceptible to be released into the environment, Directive 2001/18/EC determines the framework and principles of an environmental risk assessment (ERA) process, the results of which constitute an important section of the vaccine registration package submitted to registration authorities. In this article, we address the implications of current European regulations for the approval of live oral bacterial vaccines with emphasis on the assessment of potential risks associated with environmental release. Biosafety aspects of already registered and some promising live bacterial vaccine strains will be briefly discussed.

  12. Living Arrangements Modify the Relationship Between Depressive Symptoms and Self-care in Patients With Heart Failure.

    PubMed

    Lee, Kyoung Suk; Lennie, Terry A; Yoon, Ju Young; Wu, Jia-Rong; Moser, Debra K

    Depressive symptoms hinder heart failure patients' engagement in self-care. As social support helps improve self-care and decrease depressive symptoms, it is possible that social support buffers the negative impact of depressive symptoms on self-care. The purpose of this study is to examine the effect of living arrangements as an indicator of social support on the relationship between depressive symptoms and self-care in heart failure patients. Stable heart failure patients (N = 206) completed the Patient Health Questionnaire-9 to measure depressive symptoms. Self-care (maintenance, management, and confidence) was measured with the Self-Care of Heart Failure Index. Path analyses were used to examine associations among depressive symptoms and the self-care constructs by living arrangements. Depressive symptoms had a direct effect on self-care maintenance and management (standardized β = -0.362 and -0.351, respectively), but not on self-care confidence in patients living alone. Depressive symptoms had no direct or indirect effect on any of the 3 self-care constructs in patients living with someone. Depressive symptoms had negative effects on self-care in patients living alone, but were not related to self-care in patients living with someone. Our results suggest that negative effects of depressive symptoms on self-care are buffered by social support.

  13. Community influences on intimate partner violence in India: Women's education, attitudes towards mistreatment and standards of living.

    PubMed

    Boyle, Michael H; Georgiades, Katholiki; Cullen, John; Racine, Yvonne

    2009-09-01

    Intimate partner violence (IPV) directed towards women is a serious public health problem. Women's education may offer protection against IPV, but uncertainty exists over how it might reduce risk for IPV at the community and individual levels. The objectives of this study are to: (1) disentangle community from individual-level influences of women's education on risk for IPV; (2) quantify the moderating influence of communities on individual-level associations between women's education and IPV; (3) determine if women's attitudes towards mistreatment and living standards at the community and individual levels account for the protective influence of women's education; and (4) determine if the protective influence of education against IPV is muted among women living in communities exhibiting attitudes more accepting of mistreatment. Study information came from 68,466 married female participants in the National Family Health Survey conducted throughout India in 1998-1999. Multilevel logistic regression was used to address the study objectives. IPV showed substantial clustering at both the state (10.2%) and community levels (11.5%). At the individual level, there was a strong non-linear association between women's education and IPV, partially accounted for by household living standards. The strength of association between women's education and IPV varied from one community to the next with evidence that the acceptance of mistreatment at the community level mutes the protective influence of higher education. Furthermore, women's attitudes towards mistreatment and their standards of living accounted for community-level associations between women's education and IPV. Place of residence accounted for substantial variation in risk of IPV and also modified individual-level associations between IPV and women's education. At the community level, women's education appeared to exert much of its protective influence by altering population attitudes towards the acceptability of mistreatment. However, there was no residual association between women's education and IPV at the community level once living standards are taken into account. While women's education provides strong, independent leverage for reducing the risk of IPV, planners must keep in mind important community factors that modify its protective influence.

  14. Antibiotic Algae by Chemical Surface Engineering.

    PubMed

    Kerschgens, Isabel P; Gademann, Karl

    2018-03-02

    Chemical cell-surface engineering is a tool for modifying and altering cellular functions. Herein, we report the introduction of an antibiotic phenotype to the green alga Chlamydomonas reinhardtii by chemically modifying its cell surface. Flow cytometry and confocal microscopy studies demonstrated that a hybrid of the antibiotic vancomycin and a 4-hydroxyproline oligomer binds reversibly to the cell wall without affecting the viability or motility of the cells. The modified cells were used to inhibit bacterial growth of Gram-positive Bacillus subtilis cultures. Delivery of the antibiotic from the microalgae to the bacterial cells was verified by microscopy. Our studies provide compelling evidence that 1) chemical surface engineering constitutes a useful tool for the introduction of new, previously unknown functionality, and 2) living microalgae can serve as new platforms for drug delivery. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Modifying Exposure-Based CBT for Cambodian Refugees with Posttraumatic Stress Disorder

    ERIC Educational Resources Information Center

    Otto, Michael W.; Hinton, Devon E.

    2006-01-01

    Cambodian refugees represent a severely traumatized population living in the United States. In this paper, we describe the modification of a cognitive-behavior therapy program to facilitate delivery of an exposure-based treatment for posttraumatic stress disorder while addressing some of the challenges brought by differences in language and…

  16. Kingdom of the Tides.

    ERIC Educational Resources Information Center

    Carter, Samuel, III

    Areas of discussion are the history of tides, the forces which exert an influence upon the earth's tides, the behavior of tides as modified by terrestrial features, "freak" behavior of tides, the marine life which inhabits tidal areas, the manner in which tides have helped to shape the course of history, how tides affect our lives on a…

  17. Temperament-Based Learning Styles of Palestinian and US Children

    ERIC Educational Resources Information Center

    Oakland, Thomas; Alghorani, Mohammed Adnan; Lee, Dong Hun

    2007-01-01

    Temperament styles of 400 Palestinian children living in Gaza are described, examined for possible gender and age differences, and compared with those of 3,200 US children in light of Jung's theory of temperament as modified by Myers and Briggs. The results show that Palestinian children generally prefer practical to imaginative, feeling to…

  18. "Do You Live in a Teepee?" Aboriginal Students' Experiences with Racial Microaggressions in Canada

    ERIC Educational Resources Information Center

    Clark, D. Anthony; Kleiman, Sela; Spanierman, Lisa B.; Isaac, Paige; Poolokasingham, Gauthamie

    2014-01-01

    The purpose of the current qualitative investigation was to examine Aboriginal undergraduates' (N = 6) experiences with racial microaggressions at a leading Canadian university. The research team analyzed focus group data using a modified consensual qualitative research approach (Hill, Thompson, & Williams, 1997). The authors identified 5…

  19. Vaccine-induced canine distemper in a lesser panda.

    PubMed

    Bush, M; Montali, R J; Brownstein, D; James, A E; Appel, M J

    1976-11-01

    A fatal disease occurred in a lesser panda (Ailurus fulgens) 2 weeks after vaccination with modified live distemper vaccine. The disease clinically resembled canine distemper. Pathologically there was giant cell pneumonia, with canine distemper viral inclusion bodies in pulmonary and digestive tract epithelium. Viral isolates were indicative of an attenuated strain rather than virulent types.

  20. THE EFFECT OF HEMOPHILUS INFLUENZAE SUIS VACCINES ON SWINE INFLUENZA

    PubMed Central

    Shope, Richard E.

    1937-01-01

    Either living or heat-killed H. influenzae suis vaccines, given intramuscularly to swine, elicit an immune response capable of modifying the course of a later swine influenza infection. The protection afforded is only partial and is in no way comparable to the complete immunity afforded by swine influenza virus vaccines. PMID:19870654

  1. The Caring Community as a Context for Joining Youth Needs and Program Services.

    ERIC Educational Resources Information Center

    Ianni, Francis A. J.

    1996-01-01

    Argues that many of the needs youth have are determined by where and how they live. Suggests youth services providers should take a constructivist approach by helping communities and organizations create services that provide and nurture caring attitudes and behaviors. Presents recommendations for modifying cultures and organizing caring…

  2. Expected net benefit of vaccinating rangeland sheep against bluetongue virus using a modified-live versus killed virus vaccine

    USDA-ARS?s Scientific Manuscript database

    Recurring outbreaks of bluetongue virus in large rangeland sheep flocks in the Intermountain West of the United States have prompted questions about the economic benefits and costs of vaccinating individual flocks against bluetongue disease. We use enterprise budgets and stochastic simulation to est...

  3. Caring for Students with Type 1 Diabetes: School Nurses' Experiences

    ERIC Educational Resources Information Center

    Wang, Yueh-Ling; Volker, Deborah L.

    2013-01-01

    This qualitative study used a Husserlian phenomenological approach to obtain an understanding of the essences of five experienced Taiwanese school nurses' lived experience of caring for students with type 1 diabetes mellitus (T1DM). Audio-recorded, semi-structured, in-depth interviews were conducted. Data analysis entailed a modified method from…

  4. Inactivated and subunit vaccines against porcine reproductive and respiratory syndrome: current status and future direction

    USDA-ARS?s Scientific Manuscript database

    Within a few years of its emergence in the late 1980's, the PRRS virus had spread globally to become the foremost infectious disease concern for the pork industry. Since 1994, modified live-attenuated vaccines against porcine reproductive and respiratory syndrome virus (PRRSV-MLV) have been widely u...

  5. Punjabi Childrearing in Britain: Development of Identity, Religion and Bilingualism.

    ERIC Educational Resources Information Center

    Dosanjh, J. S.; Ghuman, Paul A. S.

    1997-01-01

    Interviewed two generations of Punjabi mothers living in Britain. Found that while second-generation Punjabis are changing some traditional mores (equal treatment of boys and girls, modified system of arranged marriage), they are also eager to transmit the core values (religion, mother tongue, familial spirit) of their culture and want their…

  6. Deletion of the thymidine kinase gene induces complete attenuation of the Georgia isolate of African swine fever virus

    USDA-ARS?s Scientific Manuscript database

    African swine fever virus (ASFV) is the etiological agent of a contagious and often lethal viral disease of domestic pigs. There are no vaccines to control Africa swine fever (ASF). Experimental vaccines have been developed using genetically modified live attenuated ASFVs obtained by specifically de...

  7. Use of Social Welfare Programs and the Disintegration of the Black Nuclear Family.

    ERIC Educational Resources Information Center

    Jewell, K. Sue

    1984-01-01

    Social welfare programs contribute to Black women's decisions to terminate their marriages by modifying existing beliefs and values, giving rise to a perception of welfare as a more viable alternative to marital relationships. But welfare is insufficient to enable the maintenance of Black women's preseparation standard of living. (RDN)

  8. Investigating the Mercalli Intensity Scale through "Lived Experience"

    ERIC Educational Resources Information Center

    Jones, Richard

    2012-01-01

    The modified Mercalli (MM) intensity scale is composed of 12 increasing levels of intensity that range from imperceptible shaking to catastrophic destruction and is designated by Roman numerals I through XII. Although qualitative in nature, it can provide a more concrete model for middle and high school students striving to understand the dynamics…

  9. Do two current canine parvovirus type 2 and 2b vaccines provide protection against the new type 2c variant?

    PubMed

    Larson, Laurie J; Schultz, R D

    2008-01-01

    Three groups (n=9 or 10) of 12-week-old canine parvovirus type 2 (CPV-2) antibody-negative puppies were vaccinated: one group with a product containing modified-live CPV-2b (Galaxy DA2PPv; Schering-Plough Animal Health), one group with a product containing modified-live CPV-2 (Continuum DAP, Intervet), and one group (controls) with sterile saline. All puppies receiving CPV-2 and CPV-2b vaccines developed antibody as determined by the hemagglutination inhibition assay. All groups of puppies were challenged with a combination of virulent CPV-2b and CPV-2c 5 weeks after vaccination. All puppies in the CPV-2 and CPV-2b vaccinated groups were protected from disease, whereas all control group puppies developed disease and 50% died or were euthanized. This study demonstrated that the CPV-2 and CPV-2b vaccine components of the Continuum DAP and Galaxy DA2PPv products, respectively, provided protection against the CPV-2b virus and also provided complete protection against the new CPV-2c variant.

  10. [Anti-influenza vaccination in animals].

    PubMed

    Bublot, M

    2009-01-01

    Until recently, Influenza was considered as a veterinary problem in avian, swine and horse only. New influenza strains able to infect and cause a disease in dogs and cats emerged these last six years. The most widely used influenza veterinary vaccines are the inactivated adjuvanted vaccines which are based on whole or split virus. New technologies have allowed the development of new generation vaccines including modified-live and vector vaccines. Modified-live influenza vaccines are available for horses only but they are in development in other species. Vector vaccines are already in use in chickens (replicative fowlpox vector) and in horses (non-replicative canarypox vector). These vaccines induce a rapid cellular and humoral immunity. Experimental studies have also shown that these vector vaccines are protective in other domestic species. These vector vaccines are compatible with the "DIVA" strategy which consists in differentiating infected from vaccinated animals and which allows disease eradication. The successive use of vector and inactivated vaccines (heterologous "prime-boost") induces a superior protective immunity in domestic poultry and constitutes a promising strategy for the control of H5N1 infection.

  11. Hemorrhagic and necrotizing hepatitis associated with administration of a modified live canine adenovirus-2 vaccine in a maned wolf (Chrysocyon brachyurus).

    PubMed

    Swenson, Julie; Orr, Kathryn; Bradley, Gregory A

    2012-06-01

    A 15-yr-old, female, maned wolf (Chrysocyon brachyurus) was euthanized after presenting semicomatose with severe, uncontrolled frank hemorrhage from her rectum 6 days following a routine physical examination and vaccination. Histopathology indicated severe hemorrhagic and necrotizing hepatitis with intranuclear basophilic inclusion bodies in the liver that were thought to be consistent with adenoviral infection. Further classification by polymerase chain reaction, immunohistochemical staining, virus isolation, and electron microscopy confirmed the etiologic agent to be canine adenovirus-2. A representative sample of the vaccine that had been used was submitted and sequenced along with the virus isolated from the maned wolf. The sequencing of the etiologic agent that had been isolated from the maned wolf was determined to be the same as the strain of virus used in the production of the modified live vaccine that had been administered 6 days prior to death. From this information, the diagnosis of vaccine-induced adenoviral hepatitis was made. This is the first confirmed case of vaccine-induced canine adenoviral hepatitis in a maned wolf.

  12. Efficacy of vaccination with La Sota strain vaccine to control Newcastle disease in village chickens in Nepal.

    PubMed

    Shrestha, Sulochana; Dhawan, Mamta; Donadeu, Meritxell; Dungu, Baptiste

    2017-02-01

    The efficacy of vaccination with Newcastle disease (ND) La Sota and R 2 B (Mukteswar) modified live strain vaccines was determined by experimental challenge and with ND La Sota vaccine under field conditions in Nepal. Booster vaccination with ND La Sota vaccine after a primary vaccination with ND La Sota vaccine, induced a geometric mean titre (GMT) of 5.0 log 2 haemagglutination inhibition (HI) units, compared to a GMT of 6.0 log 2 HI units following booster vaccination with R 2 B vaccine 1 month after primary vaccination with ND La Sota vaccine. Both vaccines provided 100% protection against challenge with a local field ND strain. Furthermore, booster vaccination with ND La Sota vaccine induced protective levels of antibody after field use in villages in Jhapa, and no outbreaks of ND occurred during the study period. The ND La Sota modified live vaccine is immunogenic and efficacious and is a suitable vaccine for use in vaccination programmes in village chickens in the rural areas of Nepal.

  13. Antigenic characterization of a formalin-inactivated poliovirus vaccine derived from live-attenuated Sabin strains.

    PubMed

    Tano, Yoshio; Shimizu, Hiroyuki; Martin, Javier; Nishimura, Yorihiro; Simizu, Bunsiti; Miyamura, Tatsuo

    2007-10-10

    A candidate inactivated poliovirus vaccine derived from live-attenuated Sabin strains (sIPV), which are used in the oral poliovirus vaccine (OPV), was prepared in a large-production scale. The modification of viral antigenic epitopes during the formalin inactivation process was investigated by capture ELISA assays using type-specific and antigenic site-specific monoclonal antibodies (MoAbs). The major antigenic site 1 was modified during the formalin inactivation of Sabin 1. Antigenic sites 1-3 were slightly modified during the formalin inactivation of Sabin 2 strain. Sites 1 and 3 were altered on inactivated Sabin 3 virus. These alterations were different to those shown by wild-type Saukett strain, used in conventional IPV (cIPV). It has been previously reported that type 1 sIPV showed higher immunogenicity to type 1 cIPV whereas types 2 and 3 sIPV induced lower level of immunogenicity than their cIPV counterparts. Our results suggest that the differences in epitope structure after formalin inactivation may account, at least in part, for the observed differences in immunogenicity between Sabin and wild-type inactivated poliovaccines.

  14. Live Music Therapy as an Active Focus of Attention for Pain and Behavioral Symptoms of Distress During Pediatric Immunization.

    PubMed

    Sundar, Sumathy; Ramesh, Bhuvaneswari; Dixit, Priyanka B; Venkatesh, Soma; Das, Prarthana; Gunasekaran, Dhandapany

    2016-07-01

    A total of 100 children coming for routine immunization to pediatric outpatient department were included and were divided into experiment (n = 50) and control (n = 50) groups. Experiment group received live music therapy during immunization procedure. Control group received no intervention. The Modified Behavior Pain Scale (MBPS), 10-point pain levels, and 10-point distress levels were documented by parents. Duration of crying was recorded by investigators. Pre- and postimmunization blood pressures and heart rates of parents holding the children were also measured and recorded by investigators. Independent and paired t tests were used for analysis. All 3 domains of the Modified Behavior Pain Scale and duration of crying showed significant improvement (P < .05) in the experiment group. Pain and distress levels also showed statistically nonsignificant improvement in experiment group. Blood pressure and heart rate of parents showed no difference. Music therapy could be helpful to children, parents, and health care providers by reducing discomfort of the child during pediatric immunization. © The Author(s) 2015.

  15. Galileo: The Added Value for Integrity in Harsh Environments.

    PubMed

    Borio, Daniele; Gioia, Ciro

    2016-01-16

    A global navigation satellite system (GNSS)-based navigation is a challenging task in a signal-degraded environments where GNSS signals are distorted by multipath and attenuated by fading effects: the navigation solution may be inaccurate or unavailable. A possible approach to improve accuracy and availability is the joint use of measurements from different GNSSs and quality check algorithms; this approach is investigated here using live GPS and Galileo signals. A modified receiver autonomous integrity monitoring (RAIM) algorithm, including geometry and separability checks, is proposed to detect and exclude erroneous measurements: the multi-constellation approach provides redundant measurements, and RAIM exploits them to exclude distorted observations. The synergy between combined GPS/Galileo navigation and RAIM is analyzed using live data; the performance is compared to the accuracy and availability of a GPS-only solution. The tests performed demonstrate that the methods developed are effective techniques for GNSS-based navigation in signal-degraded environments. The joint use of the multi-constellation approach and of modified RAIM algorithms improves the performance of the navigation system in terms of both accuracy and availability.

  16. Galileo: The Added Value for Integrity in Harsh Environments

    PubMed Central

    Borio, Daniele; Gioia, Ciro

    2016-01-01

    A global navigation satellite system (GNSS)-based navigation is a challenging task in a signal-degraded environments where GNSS signals are distorted by multipath and attenuated by fading effects: the navigation solution may be inaccurate or unavailable. A possible approach to improve accuracy and availability is the joint use of measurements from different GNSSs and quality check algorithms; this approach is investigated here using live GPS and Galileo signals. A modified receiver autonomous integrity monitoring (RAIM) algorithm, including geometry and separability checks, is proposed to detect and exclude erroneous measurements: the multi-constellation approach provides redundant measurements, and RAIM exploits them to exclude distorted observations. The synergy between combined GPS/Galileo navigation and RAIM is analyzed using live data; the performance is compared to the accuracy and availability of a GPS-only solution. The tests performed demonstrate that the methods developed are effective techniques for GNSS-based navigation in signal-degraded environments. The joint use of the multi-constellation approach and of modified RAIM algorithms improves the performance of the navigation system in terms of both accuracy and availability. PMID:26784205

  17. Targeting α-synuclein oligomers by protein-fragment complementation for drug discovery in synucleinopathies.

    PubMed

    Moussaud, Simon; Malany, Siobhan; Mehta, Alka; Vasile, Stefan; Smith, Layton H; McLean, Pamela J

    2015-05-01

    Reducing the burden of α-synuclein oligomeric species represents a promising approach for disease-modifying therapies against synucleinopathies such as Parkinson's disease and dementia with Lewy bodies. However, the lack of efficient drug discovery strategies that specifically target α-synuclein oligomers has been a limitation to drug discovery programs. Here we describe an innovative strategy that harnesses the power of bimolecular protein-fragment complementation to monitor synuclein-synuclein interactions. We have developed two robust models to monitor α-synuclein oligomerization by generating novel stable cell lines expressing α-synuclein fusion proteins for either fluorescent or bioluminescent protein-fragment complementation under the tetracycline-controlled transcriptional activation system. A pilot screen was performed resulting in the identification of two potential hits, a p38 MAPK inhibitor and a casein kinase 2 inhibitor, thereby demonstrating the suitability of our protein-fragment complementation assay for the measurement of α-synuclein oligomerization in living cells at high throughput. The application of the strategy described herein to monitor α-synuclein oligomer formation in living cells with high throughput will facilitate drug discovery efforts for disease-modifying therapies against synucleinopathies and other proteinopathies.

  18. Protection of dogs for 13 months against Bordetella bronchiseptica and canine parainfluenza virus with a modified live vaccine.

    PubMed

    Jacobs, A A C; Theelen, R P H; Jaspers, R; Horspool, L J I; Sutton, D; Bergman, J G H E; Paul, G

    2005-07-02

    Twelve specific pathogen-free (spf) puppies were vaccinated intranasally with a bivalent, modified live vaccine against infectious tracheobronchitis (group 1) and six puppies of the same age and from the same source served as unvaccinated controls (group 2). Both groups were challenged with wild-type Bordetella bronchiseptica and canine parainfluenza virus by the aerosol route 56 weeks after group 1 had been vaccinated, and at the same time six 10-week-old spf puppies from the same source (group 3) were also challenged. Oronasal swabs were taken regularly before and after the challenge, for the isolation of bacteria and viruses, and the dogs were observed for clinical signs for three weeks after the challenge. The control dogs became culture-positive for B bronchiseptica and canine parainfluenza virus, but the isolation yields from the vaccinated group were significantly lower (P<0.05). The mean clinical scores of the vaccinated group were 61 per cent lower than the scores of group 2 (P=0.009), and 90 per cent lower than the scores of group 3 (P=0.001).

  19. Efficacy of a modified live Flavobacterium columnare vaccine in fish.

    PubMed

    Shoemaker, Craig A; Klesius, Phillip H; Drennan, John D; Evans, Joyce J

    2011-01-01

    Flavobacterium columnare is an aquatic bacterium that is responsible for columnaris disease. This aquatic pathogen has a worldwide distribution and is highly infectious to both warm and cold water fish. A modified live F. columnare vaccine was developed by repeated passage of a virulent strain on increasing concentrations of rifampicin that resulted in attenuation. Here we report vaccination/challenge trials to evaluate efficacy and safety. In separate laboratory trials, immersion vaccination of channel catfish (Ictalurus punctatus) fry between 10 to 48 days post hatch (DPH) with experimental vaccine or licensed product resulted in relative percent survival (RPS) between 57-94% following challenge. Similarly, a vaccination/challenge trial using largemouth bass (Micropterus salmoides) fry at 10 DPH was performed using various doses of licensed product under laboratory conditions. Results demonstrated safety of the vaccine and significant protection following challenge with RPS values between 74-94%, depending on vaccine dose. Together, these trials demonstrate the vaccine administered to early life-stage channel catfish and largemouth bass is safe and reduces mortality following challenge with F. columnare. Published by Elsevier Ltd.

  20. Three-year serologic immunity against canine parvovirus type 2 and canine adenovirus type 2 in dogs vaccinated with a canine combination vaccine.

    PubMed

    Larson, L J; Schultz, R D

    2007-01-01

    A group of client-owned dogs and a group of dogs at a commercial kennel were evaluated for duration of antibody responses against canine parvovirus type 2 (CPV-2) and canine adenovirus type 1 (CAV-1) after receiving a combination vaccine containing recombinant canarypox-vectored canine distemper virus (CDV) and modified-live CPV-2, CAV-2, and canine parainfluenza virus, with (C6) or without (C4) two serovars of Leptospira (Recombitek C4 or C6, Merial). Duration of antibody, which correlates with protective immunity, was found to be at least 36 months in both groups. Recombitek combination vaccines can confidently be given every 3 years with assurance of protection in immunocompetent dogs against CPV-2 and CAV-1 as well as CDV. This allows this combination vaccine, like other, similar modified- live virus combination products containing CDV, CAV-2, and CPV-2, to be administered in accordance with the recommendations of the American Animal Hospital Association Canine Vaccine Task Force.

  1. Immunogenicity of a modified-live virus vaccine against bovine viral diarrhea virus types 1 and 2, infectious bovine rhinotracheitis virus, bovine parainfluenza-3 virus, and bovine respiratory syncytial virus when administered intranasally in young calves.

    PubMed

    Xue, Wenzhi; Ellis, John; Mattick, Debra; Smith, Linda; Brady, Ryan; Trigo, Emilio

    2010-05-14

    The immunogenicity of an intranasally-administered modified-live virus (MLV) vaccine in 3-8 day old calves was evaluated against bovine viral diarrhea virus (BVDV) types 1 and 2, infectious bovine rhinotracheitis (IBR) virus, parainfluenza-3 (PI-3) virus and bovine respiratory syncytial virus (BRSV). Calves were intranasally vaccinated with a single dose of a multivalent MLV vaccine and were challenged with one of the respective viruses three to four weeks post-vaccination in five separate studies. There was significant sparing of diseases in calves intranasally vaccinated with the MLV vaccine, as indicated by significantly fewer clinical signs, lower rectal temperatures, reduced viral shedding, greater white blood cell and platelet counts, and less severe pulmonary lesions than control animals. This was the first MLV combination vaccine to demonstrate efficacy against BVDV types 1 and 2, IBR, PI-3 and BRSV in calves 3-8 days of age. Copyright 2010 Elsevier Ltd. All rights reserved.

  2. Microbiome of Total Versus Live Bacteria in the Gut of Rex Rabbits

    PubMed Central

    Fu, Xiangchao; Zeng, Bo; Wang, Ping; Wang, Lihuan; Wen, Bin; Li, Ying; Liu, Hanzhong; Bai, Shiqie; Jia, Gang

    2018-01-01

    Gastrointestinal bacteria are essential for host health, and only viable microorganisms contribute to gastrointestinal functions. When evaluating the gut microbiota by next generation sequencing method, dead bacteria, which compose a proportion of gut bacteria, may distort analysis of the live gut microbiota. We collected stomach, jejunum, ileum, cecum, and colon contents from Rex rabbits. A modified propidium monoazide (PMA) treatment protocol was used to exclude DNA from dead bacteria. Analysis of untreated samples yielded total bacteria, and analysis of PMA-treated samples yielded live bacteria. Quantitative polymerase chain reaction and 16S rRNA gene sequencing were performed to evaluate the live-to-total bacteria ratio and compare the difference between live and total microbiota in the entire digestive tract. A low proportion of live bacteria in the foregut (stomach 1.12%, jejunum 1.2%, ileum 2.84%) and a high proportion of live bacteria in the hindgut (cecum 24.66%, colon 19.08%) were observed. A significant difference existed between total and live microbiota. Clostridiales, Ruminococcaceae, and S24-7 dominated the hindgut of both groups, while Acinetobacter and Cupriavidus dominated only in live foregut microbiota. Clostridiales and Ruminococcaceae abundance decreased, while S24-7 increased in live hindgut microbiota. The alpha- and beta-diversities differed significantly between groups. Analysis of networks showed the mutual relationship between live bacteria differed vastly when compared with total bacteria. Our study revealed a large number of dead bacteria existed in the digestive tract of Rex rabbits and distorted the community profile of the live microbiota. Total bacteria is an improper representation of the live gut microbiota, particularly in the foregut. PMID:29692775

  3. Food Environments and Childhood Weight Status: Effects of Neighborhood Median Income

    PubMed Central

    Sharifi, Mona; Sequist, Thomas; Block, Jason; Duncan, Dustin T.; Melly, Steven J.; Rifas-Shiman, Sheryl L.; Taveras, Elsie M.

    2015-01-01

    Abstract Background: A key aspect of any intervention to improve obesity is to better understand the environment in which decisions are being made related to health behaviors, including the food environment. Methods: Our aim was to examine the extent to which proximity to six types of food establishments is associated with BMI z-score and explore potential effect modification of this relationship. We used geographical information software to determine proximity from 49,770 pediatric patients' residences to six types of food establishments. BMI z-score obtained from the electronic health record was the primary outcome. Results: In multivariable analyses, living in closest proximity to large (β, −0.09 units; 95% confidence interval [CI], −0.13, −0.05) and small supermarkets (−0.08 units; 95% CI, −0.11, −0.04) was associated with lower BMI z-score; living in closest proximity to fast food (0.09 units; 95% CI, 0.03, 0.15) and full-service restaurants (0.07 units; 95% CI, 0.01, 0.14) was associated with a higher BMI z-score versus those living farthest away. Neighborhood median income was an effect modifier of the relationships of convenience stores and full-service restaurants with BMI z-score. In both cases, closest proximity to these establishments had more of an adverse effect on BMI z-score in lower-income neighborhoods. Conclusions: Living closer to supermarkets and farther from fast food and full-service restaurants was associated with lower BMI z-score. Neighborhood median income was an effect modifier; convenience stores and full-service restaurants had a stronger adverse effect on BMI z-score in lower-income neighborhoods. PMID:25923838

  4. Development of low level 226Ra analysis for live fish using gamma-ray spectrometry

    NASA Astrophysics Data System (ADS)

    Chandani, Z.; Prestwich, W. V.; Byun, S. H.

    2017-06-01

    A low level 226Ra analysis method for live fish was developed using a 4π NaI(Tl) gamma-ray spectrometer. In order to find out the best algorithm for accomplishing the lowest detection limit, the gamma-ray spectrum from a 226Ra point was collected and nine different methods were attempted for spectral analysis. The lowest detection limit of 0.99 Bq for an hour counting occurred when the spectrum was integrated in the energy region of 50-2520 keV. To extend 226Ra analysis to live fish, a Monte Carlo simulation model with a cylindrical fish in a water container was built using the MCNP code. From simulation results, the spatial distribution of the efficiency and the efficiency correction factor for the live fish model were determined. The MCNP model will be able to be conveniently modified when a different fish or container geometry is employed as fish grow up in real experiments.

  5. Towards physics of neural processes and behavior

    PubMed Central

    Latash, Mark L.

    2016-01-01

    Behavior of biological systems is based on basic physical laws, common across inanimate and living systems, and currently unknown physical laws that are specific for living systems. Living systems are able to unite basic laws of physics into chains and clusters leading to new stable and pervasive relations among variables (new physical laws) involving new parameters and to modify these parameters in a purposeful way. Examples of such laws are presented starting from the tonic stretch reflex. Further, the idea of control with referent coordinates is formulated and merged with the idea of hierarchical control and the principle of abundance. The notion of controlled stability of behaviors is linked to the idea of structured variability, which is a common feature across living systems and actions. The explanatory and predictive power of this approach is illustrated with respect to the control of both intentional and unintentional movements, the phenomena of equifinality and its violations, preparation to quick actions, development of motor skills, changes with aging and neurological disorders, and perception. PMID:27497717

  6. Pitheciids in fragmented habitats: Land cover change and its implications for conservation.

    PubMed

    Boyle, Sarah A

    2016-05-01

    Pitheciids (Cacajao, Callicebus, Chiropotes, and Pithecia) have experienced habitat loss and fragmentation across their geographic range in South America. Some populations living in habitat fragments live in smaller groups, travel shorter distances, and consume items that are not regularly found in the diets of populations living in continuous habitat; however, these patterns are not consistent across species. I used the IUCN Red List of Threatened Species to delineate the geographic range and conservation status of 43 pitheciid species. I calculated the amount of modified land cover within the range of each species, as well as the extent to which the remaining habitat exists in small fragments and the amount of forest lost from 2000 to 2012. Mean forest fragment size ranged from 12 to 12,027 ha, and mean forest loss from 2000 to 2012 ranged from 10.7% for Chiropotes to 0.9% for Pithecia. Critically Endangered and Endangered species represented 20.9% of the pitheciid species, and 46.5% of these species had population trends documented as decreasing. Total modified land cover was greatest for Callicebus species (18.0% of geographic range), followed by Chiropotes (13.8%), Pithecia (4.4%), and Cacajao (1.1%). Species of greater conservation concern had smaller geographic ranges, and a greater percentage of their range consisting of modified land cover than species of lower conservation concern. Species of greater conservation concern also had a greater percentage of forest lost from 2000 to 2012 and a smaller percentage of the remaining forest being protected. Most studies of pitheciids in fragments have concentrated on census data; the behavior of pitheciids in fragments has been examined for only 9 of the 43 species. Increased data on the responses of pitheciid species to forest loss and fragmentation are necessary in order to address pitheciid conservation, especially in areas undergoing severe habitat loss. © 2014 Wiley Periodicals, Inc.

  7. Versatile quantitative phase imaging system applied to high-speed, low noise and multimodal imaging (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Federici, Antoine; Aknoun, Sherazade; Savatier, Julien; Wattellier, Benoit F.

    2017-02-01

    Quadriwave lateral shearing interferometry (QWLSI) is a well-established quantitative phase imaging (QPI) technique based on the analysis of interference patterns of four diffraction orders by an optical grating set in front of an array detector [1]. As a QPI modality, this is a non-invasive imaging technique which allow to measure the optical path difference (OPD) of semi-transparent samples. We present a system enabling QWLSI with high-performance sCMOS cameras [2] and apply it to perform high-speed imaging, low noise as well as multimodal imaging. This modified QWLSI system contains a versatile optomechanical device which images the optical grating near the detector plane. Such a device is coupled with any kind of camera by varying its magnification. In this paper, we study the use of a sCMOS Zyla5.5 camera from Andor along with our modified QWLSI system. We will present high-speed live cell imaging, up to 200Hz frame rate, in order to follow intracellular fast motions while measuring the quantitative phase information. The structural and density information extracted from the OPD signal is complementary to the specific and localized fluorescence signal [2]. In addition, QPI detects cells even when the fluorophore is not expressed. This is very useful to follow a protein expression with time. The 10 µm spatial pixel resolution of our modified QWLSI associated to the high sensitivity of the Zyla5.5 enabling to perform high quality fluorescence imaging, we have carried out multimodal imaging revealing fine structures cells, like actin filaments, merged with the morphological information of the phase. References [1]. P. Bon, G. Maucort, B. Wattellier, and S. Monneret, "Quadriwave lateral shearing interferometry for quantitative phase microscopy of living cells," Opt. Express, vol. 17, pp. 13080-13094, 2009. [2] P. Bon, S. Lécart, E. Fort and S. Lévêque-Fort, "Fast label-free cytoskeletal network imaging in living mammalian cells," Biophysical journal, 106(8), pp. 1588-1595, 2014

  8. Unemployment and self-rated health: neighborhood influence.

    PubMed

    Giatti, Luana; Barreto, Sandhi M; César, Cibele C

    2010-08-01

    This work contributes to the study of the relationship between health, work and context by investigating the interaction between them in Brazil, a country with great social inequalities. It investigates whether unemployment and socioeconomic characteristics of the neighborhoods in which people live are associated with poor self-rated health after adjustment for individual sociodemographic characteristics, behavioral risk factors and health status. Moreover, it tests whether living in an area of socioeconomic deprivation modifies the association between unemployment and self-rated health. The study involved participants whose ages ranged from 15 up to 64 years, and who lived in four Brazilian cities included in the National Household Survey on Risk Behaviors and Reported Morbidity from Non-Communicable Diseases, carried out by the Ministry of Health in 2002/2003. Data from the 2000 Brazilian Population Census were used to calculate two neighborhood socioeconomic indicators: the proportion of householders with low income, a compositional variable of individual level characteristics, and residing in slums, a contextual variable not captured by individual properties. Logistic regression analysis was estimated by Generalized Estimating Equations. Of the 6426 participants, 20.6% reported poor self-rated health. Unemployment as well as residing in slums or in low income household areas were significantly associated with poor self-rated health. The magnitudes of these associations were attenuated after adjustment for sociodemographic characteristics, behavioral risk factors and other health status indicators. However, the association between unemployment and poor self-rated health was not modified by neighborhood socioeconomic indicators. Results confirm the association between unemployment and poor self-rated health, regardless of the personal or contextual characteristics studied here. Similarly, they show a clear independent association between self-rated health and neighborhood context. Even so, they do not show that the neighborhood contexts investigated modify the associations between unemployment and poor self-rated health. (c) 2010 Elsevier Ltd. All rights reserved.

  9. Snow and Rain Modify Neighbourhood Walkability for Older Adults.

    PubMed

    Clarke, Philippa; Hirsch, Jana A; Melendez, Robert; Winters, Meghan; Sims Gould, Joanie; Ashe, Maureen; Furst, Sarah; McKay, Heather

    2017-06-01

    The literature has documented a positive relationship between walkable built environments and outdoor mobility in older adults. Yet, surprisingly absent is any consideration of how weather conditions modify the impact of neighbourhood walkability. Using archived weather data linked to survey data collected from a sample of older adults in Vancouver, Canada, we found that car-dependent neighbourhoods (featuring longer block lengths, fewer intersections, and greater distance to amenities) became inaccessible in snow. Even older adults who lived in very walkable neighbourhoods walked to 25 per cent fewer destinations in snow. It is crucial to consider the impact of weather in the relationship between neighbourhood walkability and older adult mobility.

  10. Estimation of the alpha decay of Platinum isotopes using different versions of theoretical formula

    NASA Astrophysics Data System (ADS)

    Hosseini, S. S.; Hassanabadi, H.; Sobhani, H.

    The alpha decay half-lives of even-even and even-odd Platinum (Pt) nuclei have been studied within the Coulomb and proximity potential model (CPPM). The present study is restricted to even-even nuclei with A = 166-198. The results are compared with other calculations such as the Semi-empirical formula (SemFIS) from Poenaru et al. based on fission theory of alpha decay, the Viola-Seaborg (VS), Royer (R) and Brown formulae. Also, the alpha decay half-lives have been calculated using the Scaling law of Brown (SLB), the Universal Decay Law (UDL) of Qi et al., the Scaling Law of Horoi et al. (SLH), and Akrawy-Dorin formula (ADF) of Akrawy and Poenaru, which are the Royer modified formula for alpha decay half-live by adding asymmetry term.

  11. Emotional attitudes of young people completing secondary schools towards genetic modification of organisms (GMO) and genetically modified foods (GMF).

    PubMed

    Jurkiewicz, Anna; Zagórski, Jerzy; Bujak, Franciszek; Lachowski, Stanisław; Florek-Łuszczki, Magdalena

    2014-01-01

    The objective of the study was recognition of the opinions of adolescents completing secondary schools concerning genetically modified organisms and genetically modified food, especially the respondents' emotional attitude towards scientific achievements in the area of live genetically modified organisms. The study covered a group of 500 school adolescents completing secondary school at the level of maturity examination. The study was conducted by the method of a diagnostic survey using a self-designed questionnaire form. Knowledge concerning the possible health effects of consumption of food containing GMO among adolescents competing secondary schools is on a relatively low level; the adolescents examined 'know rather little' or 'very little know' about this problem. In respondents' opinions the results of reliable studies pertaining to the health effects of consumption of GMO 'rather do not exist'. The respondents are against the cultivation of GM plants and breeding of GM animals on own farm in the future. Secondary school adolescents considered that the production of genetically modified food means primarily the enrichment of biotechnological companies, higher income for food producers, and not the elimination of hunger in the world or elimination of many diseases haunting humans.

  12. Biotechnology and human rights.

    PubMed

    Feuillet-Le Mintier, B

    2001-12-01

    Biotechnology permits our world to progress. It's a tool to better apprehend the human being, but as well to let him go ahead. Applied to the living, biotechnologies present the same finality. But since their matter concerns effectively the living, they are the sources of specific dangers and particularly of that one to use the improvements obtained on the human to modify the human species. The right of the persons has to find its place to avoid that the fundamental rights of the human personality shall undergo harm. This mission assigned to the right of the persons is as so much invaluable that the economical stakes are particularly important in the domain of the biotechnologies.

  13. Biological safety concepts of genetically modified live bacterial vaccines.

    PubMed

    Frey, Joachim

    2007-07-26

    Live vaccines possess the advantage of having access to induce cell-mediated and antibody-mediated immunity; thus in certain cases they are able to prevent infection, and not only disease. Furthermore, live vaccines, particularly bacterial live vaccines, are relatively cheap to produce and easy to apply. Hence they are suitable to immunize large communities or herds. The induction of both cell-mediated immunity as well as antibody-mediated immunity, which is particularly beneficial in inducing mucosal immune responses, is obtained by the vaccine-strain's ability to colonize and multiply in the host without causing disease. For this reason, live vaccines require attenuation of virulence of the bacterium to which immunity must be induced. Traditionally attenuation was achieved simply by multiple passages of the microorganism on growth medium, in animals, eggs or cell cultures or by chemical or physical mutagenesis, which resulted in random mutations that lead to attenuation. In contrast, novel molecular methods enable the development of genetically modified organisms (GMOs) targeted to specific genes that are particularly suited to induce attenuation or to reduce undesirable effects in the tissue in which the vaccine strains can multiply and survive. Since live vaccine strains (attenuated by natural selection or genetic engineering) are potentially released into the environment by the vaccinees, safety issues concerning the medical as well as environmental aspects must be considered. These involve (i) changes in cell, tissue and host tropism, (ii) virulence of the carrier through the incorporation of foreign genes, (iii) reversion to virulence by acquisition of complementation genes, (iv) exchange of genetic information with other vaccine or wild-type strains of the carrier organism and (v) spread of undesired genes such as antibiotic resistance genes. Before live vaccines are applied, the safety issues must be thoroughly evaluated case-by-case. Safety assessment includes knowledge of the precise function and genetic location of the genes to be mutated, their genetic stability, potential reversion mechanisms, possible recombination events with dormant genes, gene transfer to other organisms as well as gene acquisition from other organisms by phage transduction, transposition or plasmid transfer and cis- or trans-complementation. For this, GMOs that are constructed with modern techniques of genetic engineering display a significant advantage over random mutagenesis derived live organisms. The selection of suitable GMO candidate strains can be made under in vitro conditions using basic knowledge on molecular mechanisms of pathogenicity of the corresponding bacterial species rather than by in vivo testing of large numbers of random mutants. This leads to a more targeted safety testing on volunteers and to a reduction in the use of animal experimentation.

  14. Health effects after firing small arms comparing leaded and unleaded ammunition.

    PubMed

    Voie, Øyvind; Borander, Anne-Katrine; Sikkeland, Liv Ingunn Bjoner; Grahnstedt, Svein; Johnsen, Arnt; Danielsen, Tor Erik; Longva, Kjetil; Kongerud, Johny

    2014-12-01

    A number of Norwegian soldiers have reported health problems after live-fire training using the HK416 rifle. The objective of this study was to characterize gaseous and particulate emissions from three different types of ammunition, and record the health effects after exposure to emissions from live-firing. Fifty-five healthy, non-smoking men (mean age 40 years) were recruited and divided randomly into three groups, one for each type of ammunition. All subjects fired the HK416 rifle in a semi-airtight tent for 60 min using leaded ammunition, unleaded ammunition and modified unleaded ammunition. Gaseous and particulate emissions were monitored within the tent. The symptoms experienced by the subjects were recorded immediately after and the day after firing using a standardized questionnaire. The concentrations of particulate matter and copper exceeded their respective occupational exposure limits (eight hours per day, five days a week) by a factor of 3 and 27, respectively. Of the 55 subjects, 54 reported general and respiratory symptoms. The total number of symptoms reported was significantly higher among shooters using unleaded ammunition as compared with the use of leaded and modified unleaded ammunition. Copper was the substance that had the highest concentration relative to its toxicity. Although the general symptoms were found to be consistent with the development of metal fume fever, the respiratory symptoms indicated an irritant effect of the airways different from that seen in metal fume fever. More symptoms were reported when unleaded ammunition was used compared with leaded and modified unleaded ammunition.

  15. Intensive management modifies soil CO2 efflux in 6-year-old Pinus taeda L. stands

    Treesearch

    Lisa J. Samuelson; Kurt Johnsen; Tom Stokes; Weinlang Lu

    2004-01-01

    Intensive forestry may reduce net CO2 emission into atmosphere by storing carbon in living biomass, dead organic matter and soil, and durable wood products. Because quantification of belowground carbon dynamics is important for reliable estimation of the carbon sequestered by intensively managed plantations, we examined soil CO2...

  16. Top 10 Research Questions Related to Physical Activity and Multiple Sclerosis

    ERIC Educational Resources Information Center

    Motl, Robert W.; Learmonth, Yvonne C.; Pilutti, Lara A.; Gappmaier, Eduard; Coote, Susan

    2015-01-01

    An estimated 2.5 million people worldwide are living with multiple sclerosis (MS), and this disease may be increasing in prevalence. MS is a disease of the central nervous system that is associated with heterogeneous symptoms and functional consequences, and the current first-line disease-modifying therapies often become ineffective later in the…

  17. Possible living flea beetle fossil in Bolivia: A new genus of flea beetles with modified hind legs (Coleoptera: Chrysomelidae: Galerucinae: Alticini)

    USDA-ARS?s Scientific Manuscript database

    A new genus (Chanealtica) with three new species (C. cuevas, C. ellimon, and C. maxi) from Bolivia is described and illustrated. It is compared with Aphthonoides Jacoby 1885, Argopistes Motschulsky 1860, Metroserrapha Bechyne 1958, Psylliodes Berthold 1827 and Psyllototus Nadein 2010. Remarkably, ba...

  18. Classical Swine Fever Outbreak after Modified Live LOM Strain Vaccination in Naive Pigs, South Korea

    PubMed Central

    Je, Sang H.; Kwon, Taeyong; Yoo, Sung J.; Lee, Dong-Uk; Lee, SeungYoon; Richt, Juergen A.

    2018-01-01

    We report classical swine fever outbreaks occurring in naive pig herds on Jeju Island, South Korea, after the introduction of the LOM vaccine strain. Two isolates from sick pigs had >99% identity with the vaccine stain. LOM strain does not appear safe; its use in the vaccine should be reconsidered. PMID:29553332

  19. Emotional Lessons: Out on the Tundra, Kids Learn To Better Understand Their Own and Others' Feelings.

    ERIC Educational Resources Information Center

    Sherman, Lee

    1999-01-01

    A nationally acclaimed antiviolence program, Second Step teaches three basic skills needed for living peacefully in society: empathy, impulse control, and anger management. In Bethel (Alaska), where student gunfire killed a student and principal in 1997, Second Step is used enthusiastically, having been modified to fit Yupik Eskimo culture and…

  20. Culture and Embodied Cognition: Moral Discourses in Internet Support Groups for Overeaters

    ERIC Educational Resources Information Center

    Ignatow, Gabriel

    2009-01-01

    This article argues that a modified version of Bourdieu's "habitus" concept can generate insights into moral culture and the ways people use culture to make changes in their lives. If revised in light of recent findings from cognitive neuroscience, the habitus allows for the analysis of culture as embodied cognitive structures linking individuals…

  1. Whole genome analysis using Bayesian models to identify candidate genes for immune response to vaccination

    USDA-ARS?s Scientific Manuscript database

    This study identified genome regions associated with variation in immune response to vaccination against bovine viral diarrhea virus type 2 (BVDV 2) in American Angus calves. Calves were born in the spring or fall of 2006-2008 (n = 620). Two doses of modified live vaccine were administered three wee...

  2. The Family Therapist's Role in Treating Families in Rural Communities: A General Systems Approach.

    ERIC Educational Resources Information Center

    Bagarozzi, Dennis A.

    1982-01-01

    Demonstrates how concepts derived from General Systems Theory can be used to understand and treat families living in impoverished, isolated rural communities in the South. Suggests interventions frequently need to be directed at the community system as a whole if community rules are to be modified for second-order change. (Author)

  3. Variations in Parenting and Adolescent Outcomes among African American and Latino Families Living in Low-Income, Urban Areas

    ERIC Educational Resources Information Center

    Roche, Kathleen M.; Ensminger, Margaret E.; Cherlin, Andrew J.

    2007-01-01

    Drawing from social disorganization theory, this study examined how perceived neighborhood conditions modified associations between parenting and delinquency, depressive symptoms, and school problem behavior among more than 800 African American and Latino 10- to 14-year-olds participating in Welfare, Children, and Families: A Three-City Study.…

  4. Achieving high aspect ratio wrinkles by modifying material network stress.

    PubMed

    Chen, Yu-Cheng; Wang, Yan; McCarthy, Thomas J; Crosby, Alfred J

    2017-06-07

    Wrinkle aspect ratio, or the amplitude divided by the wavelength, is hindered by strain localization transitions when an increasing global compressive stress is applied to synthetic material systems. However, many examples from living organisms show extremely high aspect ratios, such as gut villi and flower petals. We use three experimental approaches to demonstrate that these high aspect ratio structures can be achieved by modifying the network stress in the wrinkle substrate. We modify the wrinkle stress and effectively delay the strain localization transition, such as folding, to larger aspect ratios by using a zero-stress initial wavy substrate, creating a secondary network with post-curing, or using chemical stress relaxation materials. A wrinkle aspect ratio as high as 0.85, almost three times higher than common values of synthetic wrinkles, is achieved, and a quantitative framework is presented to provide understanding the different strategies and predictions for future investigations.

  5. A Technique for Facile and Precise Transfer of Mouse Embryos

    PubMed Central

    Sarvari, Ali; Naderi, Mohammad Mehdi; Sadeghi, Mohammad Reza; Akhondi, Mohammad Mehdi

    2013-01-01

    Background Successful Embryo Transfer (ET) technique is a fateful step of all efforts to achieve live births from in vitro produced embryos in assisted reproductive techniques or in knockout, transgenic or cloned animal projects. Small reproductive tract of mice and limitation of current techniques may not well satisfy the requirements for mass production of genetically modified mice. Genetic abnormalities of embryos, receptivity and uterine contractions, expulsion of embryos, blood, mucus or bacterial contamination on the transfer pipette tip, technical problems and even animal strain may affect embryo transfer outcome. Methods In this study, two techniques of embryo transfer in mice were compared. In conventional technique the oviduct wall was punctured with a 30-gauge needle and the loaded Pasteur pipette with embryos and medium was inserted into the hole. In new technique, embryos that were loaded in modified micropipette with minimal medium were transferred directly to the oviduct by manual piston micro-pump easily. Embryo viability was evaluated considering the percentage of live healthy newborns. Results Results of the two techniques were compared by t-test within the NPAR1WAY procedure of SAS software (ver. 9.2). The average live birth rates in the novel methods was significantly higher (42.4%) than the conventional method (21.7%, p<0.05). Conclusion In conclusion, using new embryo transfer technique improved birth rate by preventing embryos expulsion from the oviduct, saving time and easy transfer of embryos with minimum volume of medium. PMID:23626878

  6. A Nonlinear Viscoelastic Model for Ceramics at High Temperatures

    NASA Technical Reports Server (NTRS)

    Powers, Lynn M.; Panoskaltsis, Vassilis P.; Gasparini, Dario A.; Choi, Sung R.

    2002-01-01

    High-temperature creep behavior of ceramics is characterized by nonlinear time-dependent responses, asymmetric behavior in tension and compression, and nucleation and coalescence of voids leading to creep rupture. Moreover, creep rupture experiments show considerable scatter or randomness in fatigue lives of nominally equal specimens. To capture the nonlinear, asymmetric time-dependent behavior, the standard linear viscoelastic solid model is modified. Nonlinearity and asymmetry are introduced in the volumetric components by using a nonlinear function similar to a hyperbolic sine function but modified to model asymmetry. The nonlinear viscoelastic model is implemented in an ABAQUS user material subroutine. To model the random formation and coalescence of voids, each element is assigned a failure strain sampled from a lognormal distribution. An element is deleted when its volumetric strain exceeds its failure strain. Element deletion has been implemented within ABAQUS. Temporal increases in strains produce a sequential loss of elements (a model for void nucleation and growth), which in turn leads to failure. Nonlinear viscoelastic model parameters are determined from uniaxial tensile and compressive creep experiments on silicon nitride. The model is then used to predict the deformation of four-point bending and ball-on-ring specimens. Simulation is used to predict statistical moments of creep rupture lives. Numerical simulation results compare well with results of experiments of four-point bending specimens. The analytical model is intended to be used to predict the creep rupture lives of ceramic parts in arbitrary stress conditions.

  7. Effects of daily living occupational therapy and resistance exercise on the activities of daily living and muscular fitness in Guillain-Barré syndrome: a case study.

    PubMed

    Ko, Kwang-Jun; Ha, Gi-Chul; Kang, Seol-Jung

    2017-05-01

    [Purpose] The study aimed to investigate the effects of daily living occupational therapy and resistance exercise on the performance of activities of daily living and muscular fitness in a patient with Guillain-Barré syndrome. [Subject and Methods] A 35-year-old man was diagnosed with Guillain-Barré syndrome. He was hospitalized at A Hospital for 3 years, and was discharged from the hospital after he became able to execute daily life activities. After discharge, he performed daily occupational therapy and resistance exercise twice a week for 70 minutes per session for 12 weeks. Performance in the activities of daily living was assessed using the modified Barthel index, and muscular fitness was measured in terms of isokinetic muscular function using the Biodes system. [Results] The subject's Barthel index score improved from 54 points before the intervention to 62 points after 4 weeks, 69 points after 8 weeks, and 79 points after 12 weeks. In addition, his shoulder flexion and extension, knee flexion and extension, and lumbar flexion and extension were improved. [Conclusion] The present study suggests that daily living occupational therapy and resistance exercise are effective in improving the activities of daily living and muscular fitness in a patient recovering from Guillain-Barré syndrome.

  8. Animal-assisted therapy and loneliness in nursing homes: use of robotic versus living dogs.

    PubMed

    Banks, Marian R; Willoughby, Lisa M; Banks, William A

    2008-03-01

    Loneliness is a common problem in long-term care facilities (LTCF) and previous work has shown that animal-assisted therapy (AAT) can to some degree reverse loneliness. Here, we compared the ability of a living dog (Dog) and a robotic dog (AIBO) to treat loneliness in elderly patients living in LTCF. In comparison with a control group not receiving AAT, both the Dog and AIBO groups had statistically significant improvements in their levels of loneliness. As measured by a modified Lexington Attachment to Pets Scale (MLAPS), residents showed high levels of attachment to both the dog and AIBO. Subscale analysis showed that the AIBO group scored lower than the living dog on "animal rights/animal welfare" but not on "general attachment" or "people substituting." However, MLAPS measures did not correlate with changes in loneliness, showing that attachment was not the mechanism by which AAT decreases loneliness. We conclude that interactive robotic dogs can reduce loneliness in residents of LTCF and that residents become attached to these robots. However, level of attachment does not explain the decrease in loneliness associated with AAT conducted with either a living or robotic dog.

  9. Live chat alternative security protocol

    NASA Astrophysics Data System (ADS)

    Rahman, J. P. R.; Nugraha, E.; Febriany, A.

    2018-05-01

    Indonesia is one of the largest e-commerce markets in Southeast Asia, as many as 5 million people do transactions in e-commerce, therefore more and more people use live chat service to communicate with customer service. In live chat, the customer service often asks customers’ data such as, full name, address, e-mail, transaction id, which aims to verify the purchase of the product. One of the risks that will happen is sniffing which will lead to the theft of confidential information that will cause huge losses to the customer. The anticipation that will be done is build an alternative security protocol for user interaction in live chat by using a cryptographic algorithm that is useful for protecting confidential messages. Live chat requires confidentiality and data integration with encryption and hash functions. The used algorithm are Rijndael 256 bits, RSA, and SHA256. To increase the complexity, the Rijndael algorithm will be modified in the S-box and ShiftRow sections based on the shannon principle rule, the results show that all pass the Randomness test, but the modification in Shiftrow indicates a better avalanche effect. Therefore the message will be difficult to be stolen or changed.

  10. Sticky-flares for in situ monitoring of human telomerase RNA in living cells.

    PubMed

    Wu, Qilong; Liu, Zhengjie; Su, Lei; Han, Guangmei; Liu, Renyong; Zhao, Jun; Zhao, Tingting; Jiang, Changlong; Zhang, Zhongping

    2018-05-17

    Human telomerase RNA (hTR), a template of telomerase for telomeric repeat synthesis, was used to reflect the telomerase activity and act as a potential target of antitumor therapy. Here, we report a novel DNA-conjugated AuNP probe termed sticky-flares for the in situ detection of intracellular human telomerase RNA. The sticky-flares probe is capable of entering living cells directly without any auxiliary and recognizing the binding domain of human telomerase RNA. On recognition, the fluorophore-modified recognition flares can specifically bind to the target, separate from the sticky-flares and act as a fluorescent reporter to quantify and dynamically profile human telomerase RNA in living cells. We envision that the sticky-flares probe would be a valuable platform to investigate the function and regulation of hTR in antitumor therapy and hTR-related drug invention.

  11. Measurement of intrinsic physiological membrane noise in cultured living cells.

    PubMed

    Bukhari, Masroor H Shah; Miller, John H

    2010-06-01

    An experimental technique and some preliminary observations are reported here for the measurement of electric noise and potentials intrinsic to the physiological function of living cells, using an in vitro yeast cells (Saccharomyces cerevisiae) model. The design and working of technique is based on a micro-electrode-based sensor working in a modified patch-clamp configuration. We present recordings of intrinsic noise and cellular electric potentials in living and aerobically respiring cells (in an electromagnetically shielded environment). An important observation of the effect of aerobic respiration on the studied cells is discussed, whereby conspicuously higher magnitude potentials were seen with aerobically respiring active yeast cells, as compared to anaerobic or dead cells. Recorded noise potentials from aerobically respiring cells are found to have a magnitude on the order of a few microVolts/cm and fall within the range of 140- in the low-frequency (LF) band.

  12. Tuberculosis Treatment Completion Rates in Southern New Mexico Colonias.

    PubMed

    Holden, Maria Arroyo; Huttlinger, Kathleen; Schultz, Pamela; Mullins, Iris; Forster-Cox, Sue

    2016-04-01

    TB medication completion treatment rates for active TB patients living in impoverished US-Mexico border communities called colonias in southern New Mexico counties are unknown. It might be suspected that residents of colonias have lower completion rates than those living in incorporated and medically more accessible areas. A retrospective record review of closed TB case records from 1993 to 2010 of southern New Mexico border counties, was conducted using a modified version of the New Mexico Department of Health Tuberculosis Targeted Health Assessment/History form (Appendix 1). Study findings reveal that despite their unincorporated status, poorer living conditions and questionable legal status, colonia TB patients had a higher medication completion rate than their non-colonia counterparts. A robust New Mexico TB treatment program contributed to high completion rates with death being the number-one reason for treatment non-completion in both colonia and non-colonias.

  13. Functional display of ice nucleation protein InaZ on the surface of bacterial ghosts.

    PubMed

    Kassmannhuber, Johannes; Rauscher, Mascha; Schöner, Lea; Witte, Angela; Lubitz, Werner

    2017-09-03

    In a concept study the ability to induce heterogeneous ice formation by Bacterial Ghosts (BGs) from Escherichia coli carrying ice nucleation protein InaZ from Pseudomonas syringae in their outer membrane was investigated by a droplet-freezing assay of ultra-pure water. As determined by the median freezing temperature and cumulative ice nucleation spectra it could be demonstrated that both the living recombinant E. coli and their corresponding BGs functionally display InaZ on their surface. Under the production conditions chosen both samples belong to type II ice-nucleation particles inducing ice formation at a temperature range of between -5.6 °C and -6.7 °C, respectively. One advantage for the application of such BGs over their living recombinant mother bacteria is that they are non-living native cell envelopes retaining the biophysical properties of ice nucleation and do no longer represent genetically modified organisms (GMOs).

  14. Caffeine and Insomnia in People Living With HIV From the Miami Adult Studies on HIV (MASH) Cohort.

    PubMed

    Ramamoorthy, Venkataraghavan; Campa, Adriana; Rubens, Muni; Martinez, Sabrina S; Fleetwood, Christina; Stewart, Tiffanie; Liuzzi, Juan P; George, Florence; Khan, Hafiz; Li, Yinghui; Baum, Marianna K

    We explored the relationship between caffeine consumption, insomnia, and HIV disease progression (CD4+ T cell counts and HIV viral loads). Caffeine intake and insomnia levels were measured using the Modified Caffeine Consumption Questionnaire and the Pittsburgh Insomnia Rating Scale (PIRS) in 130 clinically stable participants who were living with HIV, taking antiretroviral therapy, and recruited from the Miami Adult Studies on HIV cohort. Linear regressions showed that caffeine consumption was significantly and adversely associated with distress score, quality-of-life score, and global PIRS score. Linear regression analyses also showed that global PIRS score was significantly associated with lower CD4+ T cell counts and higher HIV viral loads. Caffeine could have precipitated insomnia in susceptible people living with HIV, which could be detrimental to their disease progression states. Copyright © 2017 Association of Nurses in AIDS Care. Published by Elsevier Inc. All rights reserved.

  15. Functional display of ice nucleation protein InaZ on the surface of bacterial ghosts

    PubMed Central

    Kassmannhuber, Johannes; Rauscher, Mascha; Schöner, Lea; Witte, Angela; Lubitz, Werner

    2017-01-01

    ABSTRACT In a concept study the ability to induce heterogeneous ice formation by Bacterial Ghosts (BGs) from Escherichia coli carrying ice nucleation protein InaZ from Pseudomonas syringae in their outer membrane was investigated by a droplet-freezing assay of ultra-pure water. As determined by the median freezing temperature and cumulative ice nucleation spectra it could be demonstrated that both the living recombinant E. coli and their corresponding BGs functionally display InaZ on their surface. Under the production conditions chosen both samples belong to type II ice-nucleation particles inducing ice formation at a temperature range of between −5.6 °C and −6.7 °C, respectively. One advantage for the application of such BGs over their living recombinant mother bacteria is that they are non-living native cell envelopes retaining the biophysical properties of ice nucleation and do no longer represent genetically modified organisms (GMOs). PMID:28121482

  16. Relay kidney transplantation in Korea--legal, ethical and medical aspects.

    PubMed

    Park, Jong-Hyun; Park, Joong-Won; Koo, Young-Mo; Kim, Jang Han

    2004-07-01

    Living kidney transplantations constitute the majority of kidney transplantations in Korea. Recently, relay kidney transplantation, which is a modified form of both 'exchange transplantation' and 'living anonymous donation', has become at issue. After a living anonymous donor makes the initial donation, the next donor, who is related to the first recipient, makes the second donation; the third donor, who is related to the second recipient, makes the third donation; and so on. In relay kidney transplantation, organ trafficking, coercion of donation, assessment order, breach of agreement, and recipient burden should be evaluated with respect to ethical, legal and medical considerations. Despite these problems, a non-governmental body, the Korean Organ and Tissue Donor Program, has been promoting relay kidney transplantations to address the shortage of cadaveric kidney donations. Acceptance of the method of relay kidney transplantation requires the institution of supplementary measures to minimize the related problems.

  17. Migrating Huns and modified heads: Eigenshape analysis comparing intentionally modified crania from Hungary and Georgia in the Migration Period of Europe

    PubMed Central

    Mayall, Peter; Bitadze, Liana

    2017-01-01

    An intentionally modified head is a visually distinctive sign of group identity. In the Migration Period of Europe (4th– 7th century AD) the practice of intentional cranial modification was common among several nomadic groups, but was strongly associated with the Huns from the Carpathian Basin in Hungary, where modified crania are abundant in archaeological sites. The frequency of modified crania increased substantially in the Mtskheta region of Georgia in this time period, but there are no records that Huns settled here. We compare the Migration Period modified skulls from Georgia with those from Hungary to test the hypothesis that the Huns were responsible for cranial modification in Georgia. We use extended eigenshape analysis to quantify cranial outlines, enabling a discriminant analysis to assess group separation and identify morphological differences. Twenty-one intentionally modified skulls from Georgia are compared with sixteen from Hungary, using nineteen unmodified crania from a modern population as a comparative baseline. Results indicate that modified crania can be differentiated from modern unmodified crania with 100% accuracy. The Hungarian and Georgian crania show some overlap in shape, but can be classified with 81% accuracy. Shape gradations along the main eigenvectors indicate that the Hungarian crania show little variation in cranial shape, in accordance with a two-bandage binding technique, whereas the Georgian crania had a wider range of variation, fitting with a diversity of binding styles. As modification style is a strong signifier of social identity, our results indicate weak Hunnic influence on cranial modification in Georgia and are equivocal about the presence of Huns in Georgia. We suggest instead that other nomadic groups such as Alans and Sarmatians living in this region were responsible for modified crania in Georgia. PMID:28152046

  18. To continue, modify or relinquish driving: findings from a longitudinal study of healthy ageing.

    PubMed

    Unsworth, Carolyn A; Wells, Yvonne; Browning, Colette; Thomas, Shane A; Kendig, Hal

    2007-01-01

    The number and proportion of drivers among people entering later life continues to rise. More information on patterns of driving for older adults is required to improve service provision and traffic planning. To map the changes in driving status for a sample of drivers aged 65 years or older over the period 1994-2000, and to identify factors associated with older people continuing, modifying or relinquishing their status as drivers. The 752 participants were drawn from the Melbourne Longitudinal Studies on Healthy Ageing (MELSHA) program, a longitudinal study of people aged 65 years and older living in the community. Participants were interviewed or contacted for follow-up in 1994, 1996, 1998 and 2000 on a range of topics including their health, functional independence and driving status. Although the number of recent drivers was smaller as participants died or were admitted to nursing homes over the 6-year data collection period, relatively few participants relinquished driving while remaining in the community. Many drivers reported modifying their driving habits over time, including decisions to restrict their driving to their local area during daylight only. Relationships were explored between driving status and the key variables of age, gender, marital status, instrumental activities of daily living (IADL) independence and self-rated measures of income, health, eyesight and hearing. Multivariate analyses indicated that drivers were more likely to modify their driving habits if they were older, dependent in IADL, and rated their eyesight as poor. Similar factors predicted relinquishing driving, but in addition, women were three times more likely to relinquish driving than men (even when health and disability were taken into account) and people who rated their incomes as 'comfortable' were more likely to relinquish driving than those with lower incomes. This study confirmed previous evidence that older drivers self-regulate by modifying their driving behavior as they age. However, since few drivers voluntarily relinquish driving, further research is required to identify ways of supporting older drivers to continue to drive safely. Copyright 2007 S. Karger AG, Basel.

  19. A Cu-free clickable fluorescent probe for intracellular targeting of small biomolecules.

    PubMed

    Yamagishi, Kento; Sawaki, Kazuaki; Murata, Atsushi; Takeoka, Shinji

    2015-05-07

    We synthesized a novel cyclooctyne-based clickable fluorescent probe with versatile properties such as high cell-membrane permeability and free diffusibility in the cell. Our probe "FC-DBCO" was conjugated to an azide-modified mannose via a Cu-free click reaction in living HeLa cells and displayed intracellular specific fluorescence imaging with low background signals.

  20. Using the McDaniels Career Transitions Considerations (1991) (Modified) Form with Potential Temporary Employees.

    ERIC Educational Resources Information Center

    Preston, Debra S.

    With the many layoffs and downsizing of companies during the 1990s, many displaced workers have turned to temporary employment to earn a living while looking for permanent employment. Others have adopted "temping" as a more long-term work style. Although it may not be possible to predict whether an individual will find satisfaction or…

  1. Perception of Canadian-French Word-Final Vowels in Lexical and Morphosyntactic Minimal Pairs by Canadian English Learners of French

    ERIC Educational Resources Information Center

    Law, Franzo II

    2011-01-01

    This study investigated the perception of Canadian French word-final vowels by English-dominant and French-dominant bilinguals living in Montreal. In a modified identification task, listeners selected the response that rhymed with the target word, embedded in a carrier sentence. Minimal sets of real and nonsense target words were used, contrasting…

  2. Scientists Grow Therapeutic Protein in Engineered Soya Bean Seeds to Prevent AIDS | Poster

    Cancer.gov

    Genetically modified soya beans provide a scalable, low-cost method of producing microbicides that prevent AIDS, a technique sustainable for resource-poor countries where AIDS is spreading rapidly. According to the Joint United Nations Programme on HIV/AIDS, more than 36 million people worldwide are living with HIV. While the number of AIDS-related deaths are decreasing,

  3. Effect of Training Japanese L1 Speakers in the Production of American English /r/ Using Spectrographic Visual Feedback

    ERIC Educational Resources Information Center

    Patten, Iomi; Edmonds, Lisa A.

    2015-01-01

    The present study examines the effects of training native Japanese speakers in the production of American /r/ using spectrographic visual feedback. Within a modified single-subject design, two native Japanese participants produced single words containing /r/ in a variety of positions while viewing live spectrographic feedback with the aim of…

  4. Development and Application of Environmental Quality of Life Scale among People Residing near Three Types of Industrial Areas, Southern Thailand

    ERIC Educational Resources Information Center

    Sopsuk, Jirawan; Chongsuvivatwong, Virasakdi; Sornsrivichai, Vorasith; Hasuwanakit, Supat

    2013-01-01

    To compare QOL among rural people living in three different industrial areas and one non-industrial area in southern Thailand. A questionnaire based on the WHOQOL-BREF with environmental assessment was initially developed. After consultation with experts and pilot study, it was tested to check internal reliability and further modified as…

  5. Physical and chemical characteristics of surface fuels in masticated mixed-conifer stands of the U.S. Rocky Mountains

    Treesearch

    Robert E. Keane; Pamela G. Sikkink; Theresa B. Jain

    2018-01-01

    Mastication is a wildland fuel treatment technique that is rapidly becoming the preferred method for many fire hazard reduction projects, especially in areas where reducing fuels with prescribed fire is particularly challenging. Mastication is the process of mechanically modifying the live and dead surface and canopy biomass by chopping and shredding vegetation to...

  6. The fungus Raffaelea lauricola modifies behavior of its symbiont and vector, the redbay ambrosia beetle (Xyleborus glabratus), by altering host plant volatile production

    USDA-ARS?s Scientific Manuscript database

    The redbay ambrosia beetle, Xyleborus glabratus, is the vector of a symbiotic fungus, Raffaelea lauricola that causes laurel wilt, a highly lethal disease to members of the Lauraceae. Pioneer Xyleborus glabratus beetles infect live trees with Raffaelea lauricola, and only when trees are declining be...

  7. Neonatal mucosal immunization with a non-living, non-genetically modified Lactococcus lactis vaccine carrier induces systemic and local Th1-type immunity and protects against lethal bacterial infection

    PubMed Central

    Ramirez, Karina; Ditamo, Yanina; Rodriguez, Liliana; Picking, Wendy L.; van Roosmalen, Maarten L.; Leenhouts, Kees; Pasetti, Marcela F.

    2010-01-01

    Safe and effective immunization of newborns and infants can significantly reduce childhood mortality, yet conventional vaccines have been largely unsuccessful in stimulating the neonatal immune system. We explored the capacity of a novel mucosal antigen delivery system consisting of non-living, non-genetically modified Lactococcus lactis particles, designated Gram-positive Enhancer Matrix (GEM), to induce immune responses in the neonatal setting. Yersinia pestis LcrV, used as model protective antigen, was displayed on the GEM particles. Newborn mice immunized intranasally with GEM-LcrV developed LcrV-specific antibodies, Th1-type cell-mediated immunity, and were protected against lethal Y. pestis (plague) infection. The GEM particles activated and enhanced the maturation of neonatal dendritic cells both in vivo and in vitro. These dendritic cells showed increased capacities for secretion of pro-inflammatory and Th1-cell polarizing cytokines, antigen presentation and stimulation of CD4+ and CD8+ T cells. These data show that mucosal immunization with L. lactis GEM particles carrying vaccine antigens represents a promising approach to prevent infectious diseases early in life. PMID:19924118

  8. Vaccination with a Porcine Reproductive and Respiratory Syndrome (PRRS) Modified Live Virus Vaccine Followed by Challenge with PRRS Virus and Porcine Circovirus Type 2 (PCV2) Protects against PRRS but Enhances PCV2 Replication and Pathogenesis Compared to Results for Nonvaccinated Cochallenged Controls.

    PubMed

    Niederwerder, Megan C; Bawa, Bhupinder; Serão, Nick V L; Trible, Benjamin R; Kerrigan, Maureen A; Lunney, Joan K; Dekkers, Jack C M; Rowland, Raymond R R

    2015-12-01

    Coinfections involving porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) contribute to a group of disease syndromes known as porcine circovirus-associated disease (PCVAD). Presumably, PRRSV infection enhances PCV2 replication as a result of modulation of host immunity. The purpose of this study was to evaluate PCV2 replication and pathogenesis in pigs vaccinated with a PRRS modified live virus (MLV) vaccine and subsequently challenged with a combination of PRRSV and PCV2. During the early postchallenge period, the number of pigs with PRRSV-associated clinical signs was decreased, and average daily gain (ADG) was increased, in the vaccinated group, demonstrating the protective effect of PRRS vaccination. However, during the later postchallenge period, more pigs in the vaccinated group showed increased PCV2 viremia, decreased ADG, increased PCVAD clinical signs, and increased mortality. In this disease model, the early benefits of PRRSV vaccination were outweighed by the later amplification of PCVAD. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  9. Vaccination with a Porcine Reproductive and Respiratory Syndrome (PRRS) Modified Live Virus Vaccine Followed by Challenge with PRRS Virus and Porcine Circovirus Type 2 (PCV2) Protects against PRRS but Enhances PCV2 Replication and Pathogenesis Compared to Results for Nonvaccinated Cochallenged Controls

    PubMed Central

    Bawa, Bhupinder; Serão, Nick V. L.; Trible, Benjamin R.; Kerrigan, Maureen A.; Lunney, Joan K.; Dekkers, Jack C. M.; Rowland, Raymond R. R.

    2015-01-01

    Coinfections involving porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) contribute to a group of disease syndromes known as porcine circovirus-associated disease (PCVAD). Presumably, PRRSV infection enhances PCV2 replication as a result of modulation of host immunity. The purpose of this study was to evaluate PCV2 replication and pathogenesis in pigs vaccinated with a PRRS modified live virus (MLV) vaccine and subsequently challenged with a combination of PRRSV and PCV2. During the early postchallenge period, the number of pigs with PRRSV-associated clinical signs was decreased, and average daily gain (ADG) was increased, in the vaccinated group, demonstrating the protective effect of PRRS vaccination. However, during the later postchallenge period, more pigs in the vaccinated group showed increased PCV2 viremia, decreased ADG, increased PCVAD clinical signs, and increased mortality. In this disease model, the early benefits of PRRSV vaccination were outweighed by the later amplification of PCVAD. PMID:26446422

  10. Development of Cell-SELEX Technology and Its Application in Cancer Diagnosis and Therapy.

    PubMed

    Chen, Man; Yu, Yuanyuan; Jiang, Feng; Zhou, Junwei; Li, Yongshu; Liang, Chao; Dang, Lei; Lu, Aiping; Zhang, Ge

    2016-12-10

    SELEX (systematic evolution of ligands by exponential enrichment) is a process involving the progressive isolation of high selective ssDNA/RNA from a combinatorial single-stranded oligonucleotide library through repeated rounds of binding, partitioning and amplification. SELEX-derived single-stranded DNA/RNA molecules, called aptamers, are selected against a wide range of targets, including purified proteins, live cells, tissues, microorganisms, small molecules and so on. With the development of SELEX technology over the last two decades, various modified SELEX processes have been arisen. A majority of aptamers are selected against purified proteins through traditional SELEX. Unfortunately, more and more evidence showed aptamers selected against purified membrane proteins failed to recognize their targets in live cells. Cell-SELEX could develop aptamers against a particular target cell line to discriminate this cell line from others. Therefore, cell-SELEX has been widely used to select aptamers for the application of both diagnosis and therapy of various diseases, especially for cancer. In this review, the advantages and limitations of cell-SELEX and SELEX against purified protein will be compared. Various modified cell-SELEX techniques will be summarized, and application of cell-SELEX in cancer diagnosis and therapy will be discussed.

  11. Organically Modified Silica Nanoparticles Are Biocompatible and Can Be Targeted to Neurons In Vivo

    PubMed Central

    Kumar, Rajiv; Iacobucci, Gary J.; Kuznicki, Michelle L.; Kosterman, Andrew; Bergey, Earl J.; Prasad, Paras N.; Gunawardena, Shermali

    2012-01-01

    The application of nanotechnology in biological research is beginning to have a major impact leading to the development of new types of tools for human health. One focus of nanobiotechnology is the development of nanoparticle-based formulations for use in drug or gene delivery systems. However most of the nano probes currently in use have varying levels of toxicity in cells or whole organisms and therefore are not suitable for in vivo application or long-term use. Here we test the potential of a novel silica based nanoparticle (organically modified silica, ORMOSIL) in living neurons within a whole organism. We show that feeding ORMOSIL nanoparticles to Drosophila has no effect on viability. ORMOSIL nanoparticles penetrate into living brains, neuronal cell bodies and axonal projections. In the neuronal cell body, nanoparticles are present in the cytoplasm, but not in the nucleus. Strikingly, incorporation of ORMOSIL nanoparticles into the brain did not induce aberrant neuronal death or interfered with normal neuronal processes. Our results in Drosophila indicate that these novel silica based nanoparticles are biocompatible and not toxic to whole organisms, and has potential for the development of long-term applications. PMID:22238611

  12. Serologic response of pups to the low-passage, modified-live canine parvovirus-2 component in a combination vaccine.

    PubMed

    O'Brien, S E

    1994-04-15

    Thirty pups from a general pet population were vaccinated for canine parvovirus-2 (CPV-2), using a low-passage, modified-live CPV-2 in a combination vaccine, every 3 weeks until the pups were between 11 and 18 weeks old. Canine parvovirus-2 antibody titers were measured by means of serum neutralization (SN) immediately before each vaccination and > or = 2 weeks after final vaccination. Thirteen pups that were initially seronegative (SN titer < 2 for CPV-2) developed protective titers (SN titer > or = 16) after 1 to 3 doses of the vaccine, administered when the pups were between 8 and 17 weeks old; 11 of the 13 developed protective titers after the first dose. Seventeen pups were initially seropositive for CPV-2 (SN titer > or = 2). Of these, 8 initially had titers < 16 and 3 others had titers that fell to < 16 during the study. Nine (82%) of these 11 pups developed protective titers. Results indicated that pups at high risk for CPV-2 disease should be vaccinated until 18 to 20 weeks old, even when the low-passage vaccine is used. Pups at lower risk of CPV-2 disease should be vaccinated periodically until 16 weeks old.

  13. Development of Cell-SELEX Technology and Its Application in Cancer Diagnosis and Therapy

    PubMed Central

    Chen, Man; Yu, Yuanyuan; Jiang, Feng; Zhou, Junwei; Li, Yongshu; Liang, Chao; Dang, Lei; Lu, Aiping; Zhang, Ge

    2016-01-01

    SELEX (systematic evolution of ligands by exponential enrichment) is a process involving the progressive isolation of high selective ssDNA/RNA from a combinatorial single-stranded oligonucleotide library through repeated rounds of binding, partitioning and amplification. SELEX-derived single-stranded DNA/RNA molecules, called aptamers, are selected against a wide range of targets, including purified proteins, live cells, tissues, microorganisms, small molecules and so on. With the development of SELEX technology over the last two decades, various modified SELEX processes have been arisen. A majority of aptamers are selected against purified proteins through traditional SELEX. Unfortunately, more and more evidence showed aptamers selected against purified membrane proteins failed to recognize their targets in live cells. Cell-SELEX could develop aptamers against a particular target cell line to discriminate this cell line from others. Therefore, cell-SELEX has been widely used to select aptamers for the application of both diagnosis and therapy of various diseases, especially for cancer. In this review, the advantages and limitations of cell-SELEX and SELEX against purified protein will be compared. Various modified cell-SELEX techniques will be summarized, and application of cell-SELEX in cancer diagnosis and therapy will be discussed. PMID:27973403

  14. Immune responses to modified live virus vaccines developed from classical or highly pathogenic PRRSV following challenge with a highly pathogenic PRRSV strain.

    PubMed

    Wang, Gang; Yu, Ying; Zhang, Chong; Tu, Yabin; Tong, Jie; Liu, Yonggang; Chang, Yafei; Jiang, Chenggang; Wang, Shujie; Zhou, En-Min; Cai, Xuehui

    2016-09-01

    Modified live virus vaccines (MLVs) are used on swine farms to control porcine reproductive and respiratory syndrome virus (PRRSV). MLVs from classical PRRSV (C-PRRSV) provide some protection against emergent highly pathogenic PRRSV (HP-PRRSV). This study characterized the protective efficacy and immune response to MLVs from C-PRRSV (CH-1R) or HP-PRRSV (HuN4-F112) in a challenge using HP-PRRSV (HuN4). The outcomes were clinical signs of disease, pathological changes in the thymus and lungs, viremia, and humoral and cellular immune responses. CH-1R provided some protection against challenge with HuN4, while HuN4-F112 was protective in the HuN4 challenge. Compared to unvaccinated piglets, the vaccinated piglets had milder symptoms and fewer pathological changes in the lung and thymus. Piglets vaccinated with HuN4-F112 had higher antibody titers and lower viral loads than piglets vaccinated with CH-1R post challenge. The differences in outcome between the MLVs suggested that underlying differences in the immune responses might warrant further study. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. Solving fatigue-related problems with cardiac arrest survivors living in the community.

    PubMed

    Kim, Young Joo; Rogers, Joan C; Raina, Ketki D; Callaway, Clifton W; Rittenberger, Jon C; Leibold, Mary Lou; Holm, Margo B

    2017-09-01

    The aim was to describe fatigue-related problems reported by post-cardiac arrest adults with chronic fatigue and energy conservation strategies generated using an Energy Conservation plus Problem Solving Therapy intervention. Following an introduction to the intervention process outlined in a Participant Workbook, participants engaged in the telephone intervention by identifying one to two fatigue-related problems. They then brainstormed with the interventionist to identify potential strategies to reduce fatigue, tested them, and either modified the strategies or moved to the next problem over three to five sessions. Eighteen cardiac arrest survivors with chronic fatigue identified instrumental activities of daily living and leisure activities as fatigue-related activities more frequently than basic activities of daily living. Energy Conservation strategies used most frequently were: plan ahead, pace yourself, delegate to others, and simplify the task. Post-cardiac arrest adults living in the community with chronic fatigue can return to previous daily activities by using energy conservation strategies such as planning ahead, pacing tasks, delegating tasks, and simplifying tasks. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Towards physics of neural processes and behavior.

    PubMed

    Latash, Mark L

    2016-10-01

    Behavior of biological systems is based on basic physical laws, common across inanimate and living systems, and currently unknown physical laws that are specific for living systems. Living systems are able to unite basic laws of physics into chains and clusters leading to new stable and pervasive relations among variables (new physical laws) involving new parameters and to modify these parameters in a purposeful way. Examples of such laws are presented starting from the tonic stretch reflex. Further, the idea of control with referent coordinates is formulated and merged with the idea of hierarchical control and the principle of abundance. The notion of controlled stability of behaviors is linked to the idea of structured variability, which is a common feature across living systems and actions. The explanatory and predictive power of this approach is illustrated with respect to the control of both intentional and unintentional movements, the phenomena of equifinality and its violations, preparation to quick actions, development of motor skills, changes with aging and neurological disorders, and perception. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. Use of radiometer to reform and repair an old living house to passive solar one

    NASA Astrophysics Data System (ADS)

    Okamoto, Yoshizo; Inagaki, Terumi; Suzuki, Takakazu; Kurokawa, Takashi

    1994-03-01

    Japanese living houses mainly consist of wooden elements in high-temperature and moist conditions. To modify the hot and humid environment, a conventional old house was partially rebuilt and repaired. Especially in the winter season, a diagnostic thermographic test was used to find deteriorated and leaking parts of interior and exterior walls. Macroscopic deteriorated parts were checked again in detail. The deteriorated element was then removed. During the reconstruction process, a new solar heat and air conditioning system using a silica-gel adsorber and underground water was installed to cool and warm up the living room. Thermography tests of this remodeled house show that room temperature is always constant and mild to human beings, especially in the winter. Temperature and heat flow distribution of flowing air in the living room was measured using thermal net and wire methods. Leaking thermal streak flow of the gap was locally visualized by the IR radiometer and a highly sensitive video camera. It was verified that IR thermography is a useful measuring instrument to check thermal defects of a house.

  18. Single living predicts a higher mortality in both women and men with chronic heart failure.

    PubMed

    Mard, Shan; Nielsen, Finn Erland

    2016-09-01

    We examined the impact of single living on all-cause mortality in patients with chronic heart failure and determined if this association was modified by gender. This historical cohort study included 637 patients who were admitted to the Department of Cardiology, Herlev Hospital, Denmark, between 1 July 2005 and 30 June 2007. Baseline clinical data were obtained from patient records. Data on survival rates were obtained from the Danish Civil Registration System. Cox proportional hazard analysis was used to compute the hazard ratio (HR) of all-cause mortality, controlling for confounding factors. The median follow-up time was 2.8 years. A total of 323 (50.7%) patients died during the follow-up period. After adjustment for confounding factors, risk of death was associated with being single (HR = 1.53 (95% confidence interval: 1.19-1.96)). In a gender-stratified analysis, the risk of death did not differ among single-living women and men. Single living is a prognostic determinant of all-cause mortality in men and women with chronic heart failure. none. not relevant.

  19. Modified Separator Performing Dual Physical/Chemical Roles to Inhibit Polysulfide Shuttle Resulting in Ultrastable Li-S Batteries.

    PubMed

    Abbas, Syed Ali; Ding, Jiang; Wu, Sheng Hui; Fang, Jason; Boopathi, Karunakara Moorthy; Mohapatra, Anisha; Lee, Li Wei; Wang, Pen-Cheng; Chang, Chien-Cheng; Chu, Chih Wei

    2017-12-26

    In this paper we describe a modified (AEG/CH) coated separator for Li-S batteries in which the shuttling phenomenon of the lithium polysulfides is restrained through two types of interactions: activated expanded graphite (AEG) flakes interacted physically with the lithium polysulfides, while chitosan (CH), used to bind the AEG flakes on the separator, interacted chemically through its abundance of amino and hydroxyl functional groups. Moreover, the AEG flakes facilitated ionic and electronic transfer during the redox reaction. Live H-cell discharging experiments revealed that the modified separator was effective at curbing polysulfide shuttling; moreover, X-ray photoelectron spectroscopy analysis of the cycled separator confirmed the presence of lithium polysulfides in the AEG/CH matrix. Using this dual functional interaction approach, the lifetime of the pure sulfur-based cathode was extended to 3000 cycles at 1C-rate (1C = 1670 mA/g), decreasing the decay rate to 0.021% per cycle, a value that is among the best reported to date. A flexible battery based on this modified separator exhibited stable performance and could turn on multiple light-emitting diodes. Such modified membranes with good mechanical strength, high electronic conductivity, and anti-self-discharging shield appear to be a scalable solution for future high-energy battery systems.

  20. Establishment of a nanoparticle-assisted RT-PCR assay to distinguish field strains and attenuated strains of porcine epidemic diarrhea virus.

    PubMed

    Zhu, Yu; Wang, Gui-Hua; Cui, Yu-Dong; Cui, Shang-Jin

    2016-09-01

    Porcine epidemic diarrhea virus (PEDV) can cause serious disease and even death in neonatal piglets, resulting in serious damage to the swine industry worldwide. Open reading frame 3 (ORF3) is the only accessory gene in the PEDV genome. Previous studies have indicated that PEDV vaccine strains have a partial deletion in ORF3. In this study, a nanoparticle-assisted polymerase chain reaction (nanoparticle-assisted RT-PCR) assay targeting the ORF3 of PEDV was developed to distinguish PEDV field strains from attenuated strains by using a specific pair of primers. The PCR products of field strains and attenuated strains were 264 bp and 215 bp in length, respectively. The sensitivity and specificity of this assay were also assessed. The nanoparticle-assisted RT-PCR assay was 10-100 times more sensitive than the conventional RT-PCR assay, with no cross-reactions when amplifying porcine pseudorabies virus (PRV), porcine circovirus type 2 (PCV2), classical swine fever virus (CSFV), porcine parvovirus (PPV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine rotavirus (RV), and porcine transmissible gastroenteritis virus (TGEV). The nanoparticle-assisted RT-PCR assay we describe here can be used to distinguish field strains from vaccine strains of PEDV, and it shows promise for reducing economic loss due to PEDV infection.

  1. Prevalence of swine viral and bacterial pathogens in rodents and stray cats captured around pig farms in Korea.

    PubMed

    Truong, Quang Lam; Seo, Tae Won; Yoon, Byung-Il; Kim, Hyeon-Cheol; Han, Jeong Hee; Hahn, Tae-Wook

    2013-12-30

    In 2008, 102 rodents and 24 stray cats from the areas around 9 pig farms in northeast South Korea were used to determine the prevalence of the following selected swine pathogens: ten viral pathogens [porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), rotavirus, classical swine fever virus (CSFV), porcine circovirus type 2 (PCV2), encephalomyocarditis virus (EMCV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine parvovirus (PPV), pseudorabies virus (PRV) and Japanese encephalitis virus (JEV)] and four bacterial pathogens (Brucella, Leptospira, Salmonella and Lawsonia intracellularis). In total, 1,260 tissue samples from 102 rodents and 24 stray cats were examined by specific PCR and RT-PCR assays, including tissue samples of the brain, tonsils, lungs, heart, liver, kidneys, spleen, small intestine, large intestine and mesenteric lymph nodes. The percentages of PCR-positive rodents for the porcine pathogens were as follows: 63.7% for Leptospira, 39.2% for Brucella, 6.8% for Salmonella, 15.7% for L. intracellularis, 14.7% for PCV2 and 3.9% for EMCV. The percentages of PCR-positive stray cats for the swine pathogens were as follows: 62.5% for Leptospira, 25% for Brucella, 12.5% for Salmonella, 12.5% for L. intracellularis and 4.2% for PEDV. These results may be helpful for developing control measures to prevent the spread of infectious diseases of pigs.

  2. How does mental-physical multimorbidity express itself in lived time and space? A phenomenological analysis of encounters with depression and chronic physical illness.

    PubMed

    Coventry, Peter A; Dickens, Chris; Todd, Chris

    2014-10-01

    Mental-physical multimorbidity (the co-existence of mental and physical ill health) is highly prevalent and associated with significant impairments and high healthcare costs. While the sociology of chronic illness has developed a mature discourse on coping with long term physical illness the impact of mental and physical health have remained analytically separated, highlighting the need for a better understanding of the day-to-day complexities encountered by people living with mental-physical multimorbidity. We used the phenomenological paradigm of the lived body to elucidate how the experience of mental-physical multimorbidity shapes people's lifeworlds. Nineteen people with chronic obstructive pulmonary disease (COPD) and depression (defined as a score ≥8 on depression scale of Hospital Anxiety and Depression Scale) were recruited from secondary NHS care and interviewed at their homes. Data were analysed phenomenologically using van Manen's lifeworld existential framework of the lived body, lived time, lived space, lived relations. Additionally, we re-analysed data (using the same framework) collected from 13 people recruited from secondary NHS care with either COPD, rheumatoid arthritis, heart disease, or type 1 or type 2 diabetes and depression. The phenomenology of mental-physical multimorbidity was articulated through embodied and emotional encounters with day-to-day life in four ways: [a] participants' perception of lived time and lived space contracted; [b] time and [c] space were experienced as liminal categories, enforcing negative mood and temporal and spatial contraction; and [d] time and space could also be customised to reinstate agency and self-determination. Mental-physical multimorbidity negatively impacts on individuals' perceptions of lived time and lived space, leading to a loss of agency, heightened uncertainty, and poor well-being. Harnessing people's capacity to modify their experience of time and space may be a novel way to support people with mental-physical multimorbidity to live well with illness. Copyright © 2014 Elsevier Ltd. All rights reserved.

  3. [Knowledge and attitudes of the population about the living wills].

    PubMed

    Angel-López-Rey, Esther; Romero-Cano, Marta; Tébar-Morales, Juan Pablo; Mora-García, Cristina; Fernández-Rodríguez, Olga

    2008-01-01

    To evaluate knowledge of living wills and attitudes to these documents in the populations of two basic health areas in Toledo (Spain). We performed a descriptive, cross-sectional study in primary care centers. Participants consisted of the populations of two basic health areas selected by systematic stratified sampling. Inclusion criteria comprised age between 18 and 80 years, and absence of mental or terminal illnesses. The participants were contacted by telephone before a personal interview was carried out. Data on demographic variables and knowledge of living wills and attitudes to these documents were recorded. A total of 395 participants were interviewed (58% women) with a mean age of 46.22 years (SD: 17.13). The vast majority (88.6%) were unaware of living wills, with no significant differences in sex or religious beliefs. Most (67.8%) would sign a living will in the case of terminal illness whereas 56.3% would sign at the present moment, with no significant differences in sex or religious beliefs. More than half of the men (57.5%) would agree to modify the living will if requested by relatives versus 42.6% of the women. More than three-quarters (76.2%) believed that information on living wills was insufficient. One-third (34.5%) would allow a relative's organs to be donated even without the relative leaving express wishes, while 49.2% would donate their own organs. There was a huge lack of awareness on living wills in our population, which nevertheless proved to be highly receptive to, and in agreement with, the implications of these documents after receiving information on the topic.

  4. Hypertension and Exposure to Noise near Airports (HYENA): study design and noise exposure assessment.

    PubMed

    Jarup, Lars; Dudley, Marie-Louise; Babisch, Wolfgang; Houthuijs, Danny; Swart, Wim; Pershagen, Göran; Bluhm, Gösta; Katsouyanni, Klea; Velonakis, Manolis; Cadum, Ennio; Vigna-Taglianti, Federica

    2005-11-01

    An increasing number of people live near airports with considerable noise and air pollution. The Hypertension and Exposure to Noise near Airports (HYENA) project aims to assess the impact of airport-related noise exposure on blood pressure (BP) and cardiovascular disease using a cross-sectional study design. We selected 6,000 persons (45-70 years of age) who had lived at least 5 years near one of six major European airports. We used modeled aircraft noise contours, aiming to maximize exposure contrast. Automated BP instruments are used to reduce observer error. We designed a standardized questionnaire to collect data on annoyance, noise disturbance, and major confounders. Cortisol in saliva was collected in a subsample of the study population (n = 500) stratified by noise exposure level. To investigate short-term noise effects on BP and possible effects on nighttime BP dipping, we measured 24-hr BP and assessed continuous night noise in another subsample (n = 200). To ensure comparability between countries, we used common noise models to assess individual noise exposure, with a resolution of 1 dB(A). Modifiers of individual exposure, such as the orientation of living and bedroom toward roads, window-opening habits, and sound insulation, were assessed by the questionnaire. For four airports, we estimated exposure to air pollution to explore modifying effects of air pollution on cardiovascular disease. The project assesses exposure to traffic-related air pollutants, primarily using data from another project funded by the European Union (APMoSPHERE, Air Pollution Modelling for Support to Policy on Health and Environmental Risks in Europe).

  5. Progress towards the 'Golden Age' of biotechnology.

    PubMed

    Gartland, K M A; Bruschi, F; Dundar, M; Gahan, P B; Viola Magni, M p; Akbarova, Y

    2013-07-01

    Biotechnology uses substances, materials or extracts derived from living cells, employing 22 million Europeans in a € 1.5 Tn endeavour, being the premier global economic growth opportunity this century. Significant advances have been made in red biotechnology using pharmaceutically and medically relevant applications, green biotechnology developing agricultural and environmental tools and white biotechnology serving industrial scale uses, frequently as process feedstocks. Red biotechnology has delivered dramatic improvements in controlling human disease, from antibiotics to overcome bacterial infections to anti-HIV/AIDS pharmaceuticals such as azidothymidine (AZT), anti-malarial compounds and novel vaccines saving millions of lives. Green biotechnology has dramatically increased food production through Agrobacterium and biolistic genetic modifications for the development of 'Golden Rice', pathogen resistant crops expressing crystal toxin genes, drought resistance and cold tolerance to extend growth range. The burgeoning area of white biotechnology has delivered bio-plastics, low temperature enzyme detergents and a host of feedstock materials for industrial processes such as modified starches, without which our everyday lives would be much more complex. Biotechnological applications can bridge these categories, by modifying energy crops properties, or analysing circulating nucleic acid elements, bringing benefits for all, through increased food production, supporting climate change adaptation and the low carbon economy, or novel diagnostics impacting on personalized medicine and genetic disease. Cross-cutting technologies such as PCR, novel sequencing tools, bioinformatics, transcriptomics and epigenetics are in the vanguard of biotechnological progress leading to an ever-increasing breadth of applications. Biotechnology will deliver solutions to unimagined problems, providing food security, health and well-being to mankind for centuries to come. Copyright © 2013 Elsevier Ltd. All rights reserved.

  6. Estimating the probability of an extinction or major outbreak for an environmentally transmitted infectious disease.

    PubMed

    Lahodny, G E; Gautam, R; Ivanek, R

    2015-01-01

    Indirect transmission through the environment, pathogen shedding by infectious hosts, replication of free-living pathogens within the environment, and environmental decontamination are suspected to play important roles in the spread and control of environmentally transmitted infectious diseases. To account for these factors, the classic Susceptible-Infectious-Recovered-Susceptible epidemic model is modified to include a compartment representing the amount of free-living pathogen within the environment. The model accounts for host demography, direct and indirect transmission, replication of free-living pathogens in the environment, and removal of free-living pathogens by natural death or environmental decontamination. Based on the assumptions of the deterministic model, a continuous-time Markov chain model is developed. An estimate for the probability of disease extinction or a major outbreak is obtained by approximating the Markov chain with a multitype branching process. Numerical simulations illustrate important differences between the deterministic and stochastic counterparts, relevant for outbreak prevention, that depend on indirect transmission, pathogen shedding by infectious hosts, replication of free-living pathogens, and environmental decontamination. The probability of a major outbreak is computed for salmonellosis in a herd of dairy cattle as well as cholera in a human population. An explicit expression for the probability of disease extinction or a major outbreak in terms of the model parameters is obtained for systems with no direct transmission or replication of free-living pathogens.

  7. Gay, Orthodox, and trembling: the rise of Jewish Orthodox gay consciousness, 1970s-2000s.

    PubMed

    Ariel, Yaakov

    2007-01-01

    In 2001, the documentary movie, Trembling Before God, was played in Jewish and gay film festivals around the world, provoking strong emotional reactions. Trembling Before God comprises interviews with Orthodox Jewish gay and lesbian persons who vividly and movingly describe their struggles to live their lives as observant Jewish people, being faithful at the same time to their sexual desires and their religious tradition. Almost all the people interviewed in the movie expressed mixed emotions: love towards their tradition and attachment to their community of faith, coupled with resentment against a community, which in their eyes failed to respond with understanding to their emotional needs, thus adding to their pain. This article aims to modify the picture portrayed in the movie. The dilemmas and struggles of gays and lesbians who live their lives in Orthodox Jewish communities are indeed real. Orthodox gays and lesbians experience a greater dissonance between their sexuality and the values of their community and therefore face more anxieties and inner turmoils than gays and lesbians who live in more permissive environments. The struggles of gay and lesbian Orthodox Jews, however, are not necessarily greater than those of gays and lesbians who live their lives in other conservative communities. In fact, while it is almost impossible to be a sexually active gay or lesbian and a practicing Southern Baptist, Seventh-Day Adventist, Jehovah's Witness, or Mormon, it is not impossible for gays and lesbians to live their lives in an Orthodox Jewish environment. Amazingly, since the 1970s, thousands of gays and lesbians have given up on liberal environments and joined the ranks of traditionalist Jewish congregations.

  8. Biodegradable polymer nanocarriers for therapeutic antisense microRNA delivery in living animals

    NASA Astrophysics Data System (ADS)

    Paulmurugan, Ramasamy; Sekar, Narayana M.; Sekar, Thillai V.

    2012-03-01

    MicroRNAs are endogenous regulators of gene expression, deregulated in several cellular diseases including cancer. Altering the cellular microenvironment by modulating the microRNAs functions can regulate different genes involved in major cellular processes, and this approach is now being investigated as a promising new generation of molecularly targeted anti-cancer therapies. AntagomiRs (Antisense-miRNAs) are a novel class of chemically modified stable oligonucleotides used for blocking the functions of endogenous microRNAs, which are overexpressed. A key challenge in achieving effective microRNAbased therapeutics lies in the development of an efficient delivery system capable of specifically delivering antisense oligonucleotides and target cancer cells in living animals. We are now developing an effective delivery system designed to selectively deliver antagomiR- 21 and antagomiR-10b to triple negative breast cancer cells, and to revert tumor cell metastasis and invasiveness. The FDA-approved biodegradable PLGA-nanoparticles were selected as a carrier for antagomiRs delivery. Chemically modified antagomiRs (antagomiR-21 and antagomiR-10b) were co-encapsulated in PEGylated-PLGA-nanoparticles by using the double-emulsification (W/O/W) solvent evaporation method, and the resulting average particle size of 150-200nm was used for different in vitro and in vivo experiments. The antagomiR encapsulated PLGA-nanoparticles were evaluated for their in vitro antagomiRs delivery, intracellular release profile, and antagomiRs functional effects, by measuring the endogenous cellular targets, and the cell growth and metastasis. The xenografts of tumor cells in living mice were used for evaluating the anti-metastatic and anti-invasive properties of cells. The results showed that the use of PLGA for antagomiR delivery is not only efficient in crossing cell membrane, but can also maintain functional intracellular antagomiRs level for a extended period of time and achieve therapeutic effect in living animals.

  9. Intracellular click reaction with a fluorescent chemical Ca2+ indicator to prolong its cytosolic retention.

    PubMed

    Takei, Yoshiaki; Murata, Atsushi; Yamagishi, Kento; Arai, Satoshi; Nakamura, Hideki; Inoue, Takafumi; Takeoka, Shinji

    2013-08-25

    The powerful strategy of "intracellular click reaction" was used to retain a chemical Ca(2+) indicator in the cytosol. Specifically, a novel clickable Ca(2+) indicator "N3-fura-2 AM" was coupled with dibenzylcyclooctyl-modified biomacromolecules via copper-free click reaction in living cells and Ca(2+) oscillation was observed for an extended period of time.

  10. Evaluation of the Effectiveness of Simulation for M4 Marksmanship Training

    DTIC Science & Technology

    2014-02-01

    DEMOGRAPHIC QUESTIONNAIRE ................................................. 34 APPENDIX C: ANALYSIS OF MARKSMANSHIP PERFORMANCE DATA TO IDENTIFY POTENTIAL...machine guns and anti- armour weapons. In these simulators, firers aim a modified weapon at a target image on a screen. When the firer pulls the trigger...investigate predictors of live-fire LF6 qualification. Specifically, we examined the utility of LF6 simulator scores and trainee demographic data as

  11. Targeting Cell Surface Proteins in Molecular Photoacoustic Imaging to Detect Ovarian Cancer Early

    DTIC Science & Technology

    2013-07-01

    biology, nanotechnology, and imaging technology, molecular imaging utilizes specific probes as contrast agents to visualize cellular processes at the...This reagent was covalently coupled to the oligosaccharides attached to polypeptide side-chains of extracellular membrane proteins on living cells...website. The normal tissue gene expression profile dataset was modified and processed as described by Fang (8) and mean intensities and standard

  12. Hole-Accepting-Ligand-Modified CdSe QDs for Dramatic Enhancement of Photocatalytic and Photoelectrochemical Hydrogen Evolution by Solar Energy.

    PubMed

    Li, Xu-Bing; Liu, Bin; Wen, Min; Gao, Yu-Ji; Wu, Hao-Lin; Huang, Mao-Yong; Li, Zhi-Jun; Chen, Bin; Tung, Chen-Ho; Wu, Li-Zhu

    2016-04-01

    Solar H 2 evolution of CdSe QDs can be significantly enhanced simply by introducing a suitable hole-accepting-ligand for achieving efficient hole extraction and transfer at the nanoscale interfaces, which opens an effective pathway for dissociation of excitons to generate long-lived charge separation, thus improving the solar-to-fuel conversion efficiency.

  13. Vaccination with a porcine reproductive and respiratory syndrome modified live virus vaccine followed by challenge with PRRSV and porcine circovirus type 2 protects against PRRS but enhances PCV2 replication and parthogenesis

    USDA-ARS?s Scientific Manuscript database

    Co-infections involving porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) contribute to a group of disease syndromes known as porcine circovirus-associated disease (PCVAD). Presumably, PRRSV infection enhances PCV2 replication as a result of modulation...

  14. Organic acid blend with pure botanical product treatment reduces Escherichia coli and Salmonella populations in pure culture and in in vitro mixed ruminal microorganism fermentations

    USDA-ARS?s Scientific Manuscript database

    Foodborne pathogenic bacteria can live in the intestinal tract of food animals and can be transmitted to humans via food or indirectly through animal or fecal contact. Organic acid blend products have been used as non-antibiotic modifiers of the gastrointestinal fermentation of food animals to impr...

  15. Modeling climate and fuel reduction impacts on mixed-conifer forest carbon stocks in the Sierra Nevada, California

    Treesearch

    Matthew D. Hurteau; Timothy A. Robards; Donald Stevens; David Saah; Malcolm North; George W. Koch

    2014-01-01

    Quantifying the impacts of changing climatic conditions on forest growth is integral to estimating future forest carbon balance. We used a growth-and-yield model, modified for climate sensitivity, to quantify the effects of altered climate on mixed-conifer forest growth in the Lake Tahoe Basin, California. Estimates of forest growth and live tree carbon stocks were...

  16. "Not" Just a Matter of Style: Addressing Culturally Different Musics as Social Praxes in Secondary School Music Classes

    ERIC Educational Resources Information Center

    Goble, J. Scott

    2010-01-01

    In this article, the author aims to make evident how instructional practices could be modified or expanded in order that music education might have a "more clearly tangible and beneficial effect on the present and future lives of music learners, communities, and society at large." First, the author explains briefly how pragmatist philosophy and…

  17. Biosorption of strontium ions from simulated high-level liquid waste by living Saccharomyces cerevisiae.

    PubMed

    Qiu, Liang; Feng, Jundong; Dai, Yaodong; Chang, Shuquan

    2018-04-12

    In this study, the Saccharomyces cerevisiae (S. cerevisiae) was modified by γ-ray. The RNA-seq results reflect that the high γ-ray energies could change some gene fragments, such as deletion, recombination, and mutation. The biosorption of strontium ions (Sr 2+ ) to different types of S. cerevisiae (S. cerevisiae (K-0), modified S. cerevisiae (Y-7), and non-living S. cerevisiae (H-K)) from the simulated high-level liquid waste (S-HLLW) was assessed at different experimental conditions. The sorption experimental results show that, under an appropriate condition, γ-ray radiation can enhance its biosorption capacity slightly of Sr 2+ to S. cerevisiae. The maximum metal uptake and efficiency of Y-7 under S-HLLW were 11.656 mg g -1 and 37.91% at 32 h (wet weight), respectively. They decreased to 9.46 mg g -1 and 30.76% under radiation conditions. SEM-EDX and TEM analysis indicates that Sr 2+ was adsorbed both on the cellular surface and the inner parts of the cells. Our experimental results fit well to the Langmuir and Freundlich model isotherms (r 2  > 0.94), and the maximum biosorption capacity values reached q max  > 24.74 mg g -1 at 32 °C. Negative values of ΔG 0 and positive values of ΔH 0 were observed, indicating the spontaneous and endothermic nature of Sr 2+ biosorption on modified S. cerevisiae. The biosorption kinetics follow a pseudo-second-order equation at 32 °C (r 2  > 0.94). The desorption efficiency of Sr 2+ adsorbed onto Y-7 was 7.65 ± 0.52%, 76.51 ± 2.13%, and 65.62 ± 2.42% by deionized water, 1 M HCl, and 0.1 M EDTA-Na, respectively. However, they were lower than H-K (18.82, 83.32, and 73.32%). Our findings demonstrate that living S. cerevisiae (Y-7) is a promising sorbent material for the treatment of radioactive process streams.

  18. Physical performance tests after stroke: reliability and validity.

    PubMed

    Maeda, A; Yuasa, T; Nakamura, K; Higuchi, S; Motohashi, Y

    2000-01-01

    To evaluate the reliability and validity of the modified physical performance tests for stroke survivors who live in a community. The subjects included 40 stroke survivors and 40 apparently healthy independent elderly persons. The physical performance tests for the stroke survivors comprised two physical capacity evaluation tasks that represented physical abilities necessary to perform the main activities of daily living, e.g., standing-up ability (time needed to stand up from bed rest) and walking ability (time needed to walk 10 m). Regarding the reliability of tests, significant correlations were confirmed between test and retest of physical performance tests with both short and long intervals in individuals after stroke. Regarding the validity of tests, the authors studied the significant correlations between the maximum isometric strength of the quardriceps muscle and the time needed to walk 10 m, centimeters reached while sitting and reaching, and the time needed to stand up from bed rest. The authors confirmed that there were significant correlations between the instrumental activity of daily living and the time needed to stand up from bed rest, along with the time needed to walk 10 m for the stroke survivors. These physical performance tests are useful guides for evaluating a level of activity of daily living and physical frailty of stroke survivors living in a community.

  19. Combining bio-electrospraying with gene therapy: a novel biotechnique for the delivery of genetic material via living cells.

    PubMed

    Ward, Eliot; Chan, Emma; Gustafsson, Kenth; Jayasinghe, Suwan N

    2010-05-01

    The investigations reported in this article demonstrate the ability of bio-electrosprays and cell electrospinning to deliver a genetic construct in association with living cells. Previous studies on both bio-electrosprays and cell electrospinning demonstrated great promise for tissue engineering and regenerative biology/medicine. The investigations described herein widen the applicability of these biotechniques by combining gene therapy protocols, resulting in a novel drug delivery methodology previously unexplored. In these studies a human cell line was transduced with recombinant self-inactivating lentiviral particles. These particles incorporated a green fluorescent protein fused to an endosomal targeting construct. This construct encodes a peptide, which can subsequently be detected on the surface of cells by specific T-cells. The transduced cell line was subsequently manipulated in association with either bio-electrospraying or cell electrospinning. Hence this demonstrates (i) the ability to safely handle genetically modified living cells and (ii) the ability to directly form pre-determined architectures bearing living therapeutic cells. This merged technology demonstrates a unique approach for directly forming living therapeutic architectures for controlled and targeted release of experimental cells/genes, as well as medical cell/gene therapeutics for a plethora of biological and medical applications. Hence, such developments could be applied to personalised medicine.

  20. Engineering and exploitation of a fluorescent HIV-1 gp120 for live cell CD4 binding assays

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Costantini, Lindsey M.; Irvin, Susan C.; Department of Microbiology and Immunology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461

    The HIV-1 envelope glycoprotein, gp120, binds the host cell receptor, CD4, in the initial step of HIV viral entry and infection. This process is an appealing target for the development of inhibitory drugs and neutralizing antibodies. To study gp120 binding and intracellular trafficking, we engineered a fluorescent fusion of the humanized gp120 JRFL HIV-1 variant and GFP. Gp120-sfGFP is glycosylated with human sugars, robustly expressed, and secreted from cultured human cells. Protein dynamics, quality control, and trafficking can be visualized in live cells. The fusion protein can be readily modified with different gp120 variants or fluorescent proteins. Finally, secreted gp120-sfGFPmore » enables a sensitive and easy binding assay that can quantitatively screen potential inhibitors of gp120-CD4 binding on live cells via fluorescence imaging or laser scanning cytometry. This adaptable research tool should aid in studies of gp120 cell biology and the development of novel anti-HIV drugs. - Highlights: • Development of fluorescent protein labeled HIV-1 envelope gp120. • Imaging of gp120 dynamics and trafficking in live cells. • Quantitative visual assay of antibody-mediated inhibition of gp120 binding to CD4 on live cells.« less

  1. Promoting Cas9 degradation reduces mosaic mutations in non-human primate embryos

    PubMed Central

    Tu, Zhuchi; Yang, Weili; Yan, Sen; Yin, An; Gao, Jinquan; Liu, Xudong; Zheng, Yinghui; Zheng, Jiezhao; Li, Zhujun; Yang, Su; Li, Shihua; Guo, Xiangyu; Li, Xiao-Jiang

    2017-01-01

    CRISPR-Cas9 is a powerful new tool for genome editing, but this technique creates mosaic mutations that affect the efficiency and precision of its ability to edit the genome. Reducing mosaic mutations is particularly important for gene therapy and precision genome editing. Although the mechanisms underlying the CRSIPR/Cas9-mediated mosaic mutations remain elusive, the prolonged expression and activity of Cas9 in embryos could contribute to mosaicism in DNA mutations. Here we report that tagging Cas9 with ubiquitin-proteasomal degradation signals can facilitate the degradation of Cas9 in non-human primate embryos. Using embryo-splitting approach, we found that shortening the half-life of Cas9 in fertilized zygotes reduces mosaic mutations and increases its ability to modify genomes in non-human primate embryos. Also, injection of modified Cas9 in one-cell embryos leads to live monkeys with the targeted gene modifications. Our findings suggest that modifying Cas9 activity can be an effective strategy to enhance precision genome editing. PMID:28155910

  2. Assessing the impact of the Indian Ocean tsunami on households: a modified domestic assets index approach.

    PubMed

    Arlikatti, Sudha; Peacock, Walter Gillis; Prater, Carla S; Grover, Himanshu; Sekar, Arul S Gnana

    2010-07-01

    This paper offers a potential measurement solution for assessing disaster impacts and subsequent recovery at the household level by using a modified domestic assets index (MDAI) approach. Assessment of the utility of the domestic assets index first proposed by Bates, Killian and Peacock (1984) has been confined to earthquake areas in the Americas and southern Europe. This paper modifies and extends the approach to the Indian sub-continent and to coastal surge hazards utilizing data collected from 1,000 households impacted by the Indian Ocean tsunami (2004) in the Nagapattinam district of south-eastern India. The analyses suggest that the MDAI scale is a reliable and valid measure of household living conditions and is useful in assessing disaster impacts and tracking recovery efforts over time. It can facilitate longitudinal studies, encourage cross-cultural, cross-national comparisons of disaster impacts and inform national and international donors of the itemized monetary losses from disasters at the household level.

  3. Apparatus and method for modifying the operation of a robotic vehicle in a real environment, to emulate the operation of the robotic vehicle operating in a mixed reality environment

    DOEpatents

    Garretson, Justin R [Albuquerque, NM; Parker, Eric P [Albuquerque, NM; Gladwell, T Scott [Albuquerque, NM; Rigdon, J Brian [Edgewood, NM; Oppel, III, Fred J.

    2012-05-29

    Apparatus and methods for modifying the operation of a robotic vehicle in a real environment to emulate the operation of the robotic vehicle in a mixed reality environment include a vehicle sensing system having a communications module attached to the robotic vehicle for communicating operating parameters related to the robotic vehicle in a real environment to a simulation controller for simulating the operation of the robotic vehicle in a mixed (live, virtual and constructive) environment wherein the affects of virtual and constructive entities on the operation of the robotic vehicle (and vice versa) are simulated. These effects are communicated to the vehicle sensing system which generates a modified control command for the robotic vehicle including the effects of virtual and constructive entities, causing the robot in the real environment to behave as if virtual and constructive entities existed in the real environment.

  4. Expanding the chemical toolbox for the synthesis of large and uniquely modified proteins

    NASA Astrophysics Data System (ADS)

    Bondalapati, Somasekhar; Jbara, Muhammad; Brik, Ashraf

    2016-05-01

    Methods to prepare proteins that include a specific modification at a desired position are essential for understanding their cellular functions and physical properties in living systems. Chemical protein synthesis, which relies on the chemoselective ligation of unprotected peptides, enables the preparation of modified proteins that are not easily fabricated by other methods. In contrast to recombinant approaches, chemical synthesis can be used to prepare protein analogues such as D-proteins, which are useful in protein structure determination and the discovery of novel therapeutics. Post-translationally modifying proteins is another example where chemical protein synthesis proved itself as a powerful approach for preparing samples with high homogeneity and in workable quantities. In this Review, we discuss the basic principles of the field, focusing on novel chemoselective peptide ligation approaches such as native chemical ligation and the recent advances based on this method with a proven record of success in the synthesis of highly important protein targets.

  5. The roles of ubiquitin modifying enzymes in neoplastic disease.

    PubMed

    Kumari, Nishi; Jaynes, Patrick William; Saei, Azad; Iyengar, Prasanna Vasudevan; Richard, John Lalith Charles; Eichhorn, Pieter Johan Adam

    2017-12-01

    The initial experiments performed by Rose, Hershko, and Ciechanover describing the identification of a specific degradation signal in short-lived proteins paved the way to the discovery of the ubiquitin mediated regulation of numerous physiological functions required for cellular homeostasis. Since their discovery of ubiquitin and ubiquitin function over 30years ago it has become wholly apparent that ubiquitin and their respective ubiquitin modifying enzymes are key players in tumorigenesis. The human genome encodes approximately 600 putative E3 ligases and 80 deubiquitinating enzymes and in the majority of cases these enzymes exhibit specificity in sustaining either pro-tumorigenic or tumour repressive responses. In this review, we highlight the known oncogenic and tumour suppressive effects of ubiquitin modifying enzymes in cancer relevant pathways with specific focus on PI3K, MAPK, TGFβ, WNT, and YAP pathways. Moreover, we discuss the capacity of targeting DUBs as a novel anticancer therapeutic strategy. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. 3D conformal radiation therapy for palliative treatment of canine nasal tumors.

    PubMed

    Buchholz, Julia; Hagen, Regine; Leo, Chiara; Ebling, Alessia; Roos, Malgorzata; Kaser-Hotz, Barbara; Bley, Carla Rohrer

    2009-01-01

    We evaluated the response of 38 dogs treated with a coarsely fractionated, palliative radiation protocol based on CT-based 3D treatment planning. Dogs with histologically confirmed malignant nasal tumors were studied. Treatment prescriptions consisted of 3-4 x 8 Gy, 4-5 x 6 Gy, or 10 x 3 Gy fractions. Selected patient and tumor factors were evaluated for an effect on outcome. Resolution of clinical signs was reported after irradiation in all dogs. Acute toxicities were mild and short lived. Thirty-seven of 38 dogs died or were euthanized due to tumor-related disease. Overall median progression-free interval (PFI) was 10 months. Tumor stage affected response, with modified stage 1 patients having a median PFI 21.3 months vs. a median PFI of 8.5 months for modified stage 2 patients (P = 0.0006). Modified stage was the only factor significantly related to outcome. Based on these findings, a palliative radiation prescription based on computerized treatment planning may be justified in some canine nasal tumor patients.

  7. Morphological characteristics of developmental stages of Acanthamoeba and Naegleria species before and after staining by various techniques.

    PubMed

    Ithoi, Init; Ahmad, Arine-Fadzlun; Mak, J W; Nissapatorn, Veeranoot; Lau, Yee-Ling; Mahmud, Rohela

    2011-11-01

    Seven stains were studied to determine the best color and contrast for staining the developmental stages of free living pathogenic Acanthamoeba and Naegleria species. The acid-fast bacilli stain (AFB) produced a blue color without contrast; trichrome-eosin and modified Field's showed various color contrasts; Giemsa, iron-hematoxylin, modified AFB and Gram produced only one color which distinguished the nucleus, nucleolus, cytoplasm, food- and water-vacuoles. The motile organs (acanthopodia, pseudopodia, lobopodia and flagella) were also clearly differentiated but produced a similar color as the cytoplasm. These motile organelles were first induced by incubating at 37 degrees C for at least 15 minutes and then fixing with methanol in order to preserve the protruding morphology prior to staining. The trichrome-eosin and iron-hematoxylin stains showed good color contrast for detecting all three stages, the trophozoite, cyst and flagellate; Giemsa and Gram stained the trophozoite and flagellate stages; the modified Field's and modified AFB stains stained only the trophozoite stage. Depending on the purpose, all these stains (except the AFB stain) can be used to identify the developmental stages of Acanthamoeba and Naegleria for clinical, epidemiological or public health use.

  8. The analysis of predictability of recent alpha decay formulae and the alpha partial half-lives of some exotic nuclei

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Dasgupta-Schubert, N.; Reyes, M. A.; Tamez, V. A.

    2009-04-20

    Alpha decay is one of the two main decay modes of the heaviest nuclei, (SHE), and constitutes one of the dominant decay modes of highly neutron deficient medium mass nuclei ('exotics'). Thus identifying and characterizing the alpha decay chains form a crucial part of the identification of SHE. We report the extension of the previously developed method for the detailed and systematic investigation of the reliability of the three main extant analytical formulae of alpha decay half-lives: the generalized liquid drop model based formula of Royer et al. (FR), the Sobiczewski modified semi-empirical Viola-Seaborg formula (VSS) and the recent phenomenologicalmore » formula of Sobiczewski and Parkhomenko (SP)« less

  9. Pattern and prevalence of smoking among students at King Faisal University, Al Hassa, Saudi Arabia.

    PubMed

    Al-Mohamed, H I; Amin, T T

    2010-01-01

    The study describes the prevalence of different forms of smoking, and the correlates of current smoking, by male students of King Faisal University, Saudi Arabia. A random sample of 1382 students at 9 colleges answered a self-administered questionnaire based on the Global Youth Tobacco Survey plus the modified Fagerström Test for Nicotine Dependence. The prevalence of current smoking was 28.1% (21.6% for cigarettes, 14.6% for waterpipe). Of current smokers, 41.4% were living in homes where others smoke and 17.0% initiated smoking below age 12 years. In logistic regression analysis older age, living away from home, smoking by family and close friends and exposure to tobacco promotion were predictors of current smoking status.

  10. Periscope for noninvasive two-photon imaging of murine retina in vivo

    PubMed Central

    Stremplewski, Patrycjusz; Komar, Katarzyna; Palczewski, Krzysztof; Wojtkowski, Maciej; Palczewska, Grazyna

    2015-01-01

    Two-photon microscopy allows visualization of subcellular structures in the living animal retina. In previously reported experiments it was necessary to apply a contact lens to each subject. Extending this technology to larger animals would require fitting a custom contact lens to each animal and cumbersome placement of the living animal head on microscope stage. Here we demonstrate a new device, periscope, for coupling light energy into mouse eye and capturing emitted fluorescence. Using this periscope we obtained images of the RPE and their subcellular organelles, retinosomes, with larger field of view than previously reported. This periscope provides an interface with a commercial microscope, does not require contact lens and its design could be modified to image retina in larger animals. PMID:26417507

  11. [Medicine behind bars. Illness is usual not grounds for modifying imprisonment].

    PubMed

    Schwarzkopf, A; Zenker, M

    2002-05-16

    The article describes the possibilities of medical care in prison as exemplified by the Kassel 1 prison with its attached central hospital facility. The main areas of medical care covered there are the treatment of wounds, conservative treatment of fractures, ophthalmology, ENT, urology, dentistry and, via external consultant physicians, also internal medicine, pneumology, dermatology and gynecology. Such infections as hepatitis B and C, syphilis, and tuberculosis have a greater prevalence among prisoners--in contrast to other infections afflicting people housed under similar living conditions, such as in communal living facilities, which show no such increased prevalence. Furthermore, there is a relatively high percentage of injuries, including those that are self-inflicted. The problem of certifying a prisoner medically unfit to tolerate imprisonment is discussed.

  12. Modified cleaning method for biomineralized components

    NASA Astrophysics Data System (ADS)

    Tsutsui, Hideto; Jordan, Richard W.

    2018-02-01

    The extraction and concentration of biomineralized components from sediment or living materials is time consuming and laborious and often involves steps that remove either the calcareous or siliceous part, in addition to organic matter. However, a relatively quick and easy method using a commercial cleaning fluid for kitchen drains, sometimes combined with a kerosene soaking step, can produce remarkable results. In this study, the method is applied to sediments and living materials bearing calcareous (e.g., coccoliths, foraminiferal tests, holothurian ossicles, ichthyoliths, and fish otoliths) and siliceous (e.g., diatom valves, silicoflagellate skeletons, and sponge spicules) components. The method preserves both components in the same sample, without etching or partial dissolution, but is not applicable to unmineralized components such as dinoflagellate thecae, tintinnid loricae, pollen, or plant fragments.

  13. Design and validation of a new ratiometric intracellular pH imaging probe using lanthanide-doped upconverting nanoparticles.

    PubMed

    Du, Shuoren; Hernández-Gil, Javier; Dong, Hao; Zheng, Xiaoyu; Lyu, Guangming; Bañobre-López, Manuel; Gallo, Juan; Sun, Ling-Dong; Yan, Chun-Hua; Long, Nicholas J

    2017-10-17

    pH homeostasis is strictly controlled at a subcellular level. A deregulation of the intra/extra/subcellular pH environment is associated with a number of diseases and as such, the monitoring of the pH state of cells and tissues is a valuable diagnostic tool. To date, only a few tools have been developed to measure the pH in living cells with the spatial resolution needed for intracellular imaging. Among the techniques available, only optical imaging offers enough resolution and biocompatibility to be proposed for subcellular pH monitoring. We present herein a ratiometric probe based on upconversion nanoparticles modified with a pH sensitive moiety for the quantitative imaging of pH at the subcellular level in living cells. This system provides the properties required for live cell quantitative imaging i.e. positive cellular uptake, biocompatibility, long wavelength excitation, sensitive response to pH within a biologically relevant range, and self-referenced signal.

  14. Strategies and Considerations for Distributing and Recovering Mouse Lines

    PubMed Central

    Du, Yubin; Xie, Wen; Liu, Chengyu

    2012-01-01

    As more and more genetically modified mouse lines are being generated, it becomes increasingly common to share animal models among different research institutions. Live mice are routinely transferred between animal facilities. Due to various issues concerning animal welfare, intellectual property rights, colony health status and biohazard, significant paperwork and coordination are required before any animal travel can take place. Shipping fresh or frozen preimplantation embryos, gametes, or reproductive organs can bypass some of the issues associated with live animal transfer, but it requires the receiving facilities to be able to perform delicate and sometimes intricate procedures such as embryo transfer, in vitro fertilization (IVF), or ovary transplantation. Here, we summarize the general requirements for live animal transport and review some of the assisted reproductive technologies (ART) that can be applied to shipping and reviving mouse lines. Intended users of these methods should consult their institution’s responsible official to find out whether each specific method is legal or appropriate in their own animal facilities. PMID:20691859

  15. Preventing falls in assisted living: Results of a quality improvement pilot study.

    PubMed

    Zimmerman, Sheryl; Greene, Angela; Sloane, Philip D; Mitchell, Madeline; Giuliani, Carol; Nyrop, Kirsten; Walsh, Edith

    Residents of assisted living (AL) communities are at high risk for falls, which result in negative outcomes and high health care costs. Adapting effective falls prevention programs for AL quality improvement (QI) has the potential to reduce falls, improve resident quality of life, and reduce costs. This project tested the feasibility and outcomes of an evidence-based multi-component QI program, the Assisted Living Falls Prevention and Monitoring Program (AL-FPMP). Resident posture and gait improved, likely due to exercise and/or physical therapy. Effective falls prevention QI programs can be implemented in AL, and are advised to (1) establish and maintain a falls team to create a culture focused on the reduction of falls risk; (2) teach staff to assess residents using the Morse Falls Scale to increase their awareness of residents' falls risk and improvement; and (3) modify existing exercise programs to address balance and lower body strength. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Social media and organ donation: Ethically navigating the next frontier.

    PubMed

    Henderson, M L; Clayville, K A; Fisher, J S; Kuntz, K K; Mysel, H; Purnell, T S; Schaffer, R L; Sherman, L A; Willock, E P; Gordon, E J

    2017-11-01

    As the organ shortage continues to grow, the creation of social media communities by transplant hospitals and the public is rapidly expanding to increase the number of living donors. Social media communities are arranged in myriad ways and without standardization, raising concerns about transplant candidates' and potential donors' autonomy and quality of care. Social media communities magnify and modify extant ethical issues in deceased and living donation related to privacy, confidentiality, professionalism, and informed consent, and increase the potential for undue influence and coercion for potential donors and transplant candidates. Currently, no national ethical guidelines have been developed in the United States regarding the use of social media to foster organ transplantation. We provide an ethical framework to guide transplant stakeholders in using social media for public and patient communication about transplantation and living donation, and offer recommendations for transplant clinical practice and future research. © 2017 The American Society of Transplantation and the American Society of Transplant Surgeons.

  17. Within-season increase in parental investment in a long-lived bird species: investment shifts to maximize successful reproduction?

    PubMed

    Schneider, N A; Griesser, M

    2015-01-01

    In nest-building species predation of nest contents is a main cause of reproductive failure and parents have to trade off reproductive investment against antipredatory behaviours. While this trade-off is modified by lifespan (short-lived species prioritize current reproduction; long-lived species prioritize future reproduction), it may vary within a breeding season, but this idea has only been tested in short-lived species. Yet, life history theory does not make any prediction how long-lived species should trade off current against future reproductive investment within a season. Here, we investigated this trade-off through predator-exposure experiments in a long-lived bird species, the brown thornbill. We exposed breeding pairs that had no prior within-season reproductive success to the models of a nest predator and a predator of adults during their first or second breeding attempt. Overall, parents reduced their feeding rate in the presence of a predator, but parents feeding second broods were more risk sensitive and almost ceased feeding when exposed to both types of predators. However, during second breeding attempts, parents had larger clutches and a higher feeding rate in the absence of predators than during first breeding attempts and approached both types of predators closer when mobbing. Our results suggest that the trade-off between reproductive investment and risk-taking can change in a long-lived species within a breeding season depending on both prior nest predation and renesting opportunities. These patterns correspond to those in short-lived species, raising the question of whether a within-season shift in reproductive investment trade-offs is independent of lifespan. © 2014 European Society For Evolutionary Biology. Journal of Evolutionary Biology © 2014 European Society For Evolutionary Biology.

  18. Impact of estradiol valerate/dienogest on work productivity and activities of daily living in women with heavy menstrual bleeding.

    PubMed

    Wasiak, Radoslaw; Filonenko, Anna; Vanness, David J; Law, Amy; Jeddi, Mark; Wittrup-Jensen, Kim U; Stull, Donald E; Siak, Steven; Jensen, Jeffrey T

    2013-04-01

    To quantify the change in work productivity and activities of daily living in North American women with heavy menstrual bleeding (HMB) treated with estradiol valerate/dienogest (E2V/DNG; Qlaira(®)/Natazia(®)) compared to placebo. Women in the United States and Canada, aged 20-53 years with an objective diagnosis of HMB and no recognizable anatomical pathology, were treated with E2V/DNG or placebo for seven cycles (196 days). Main outcome measures included work productivity (i.e., productivity while at work) and activities of daily living measured using a modified Work Productivity and Activity Impairment Questionnaire (mWPAI) on a Likert scale from 0 to 10 (higher values denote higher impairment levels). In both countries, significant improvement was observed between baseline and end of treatment in work productivity and activities of daily living impairment. The improvements in work productivity and activities of daily living with E2V/DNG treatment relative to placebo ranged from 37.2% to 39.2% across both countries. Monthly gains due to E2V/DNG treatment (net of placebo improvement) associated with improvement in work productivity were estimated to be US$80.2 and Can$70.8 (US$58.5) and those associated with improvement in activities of daily living were estimated to be US$84.9 and Can$73.5 (US$60.7). E2V/DNG was shown to have a consistent positive impact on work productivity and activities of daily living in U.S. and Canadian women with HMB. In addition, these improvements in work productivity and activities of daily living were associated with a reduction in HMB-related monetary burden compared to the placebo group.

  19. Maintaining Genome Stability: The Role of Helicases and Deaminases

    DTIC Science & Technology

    2008-07-01

    deaminases. The cells response to different forms of damage is fundamental to its ability to repair itself when challenged by environmental or...Page 4 of 12 INTRODUCTION Genomic DNA stores all the information for living organisms. The faithful duplication the maintanance of DNA are...maturation of the immune system by modifying enzymes called deaminases. The cells response to different forms of damage is fundamental to its ability to

  20. Studies on chemical evolution using carbon suboxide

    NASA Technical Reports Server (NTRS)

    Yanagawa, H.; Egami, F.

    1980-01-01

    It was learned that when carbon suboxide polymers react with hydroxylamine in modified sea water the amino acids glycine and lysine are produced and that they react with urea to produce nucleic acid-like substances. These results lead to the supposition that it is possible that carbon suboxide polymers may have accumulated in the primeval sea some three or more billion years ago and were transformed into the procursors of living molecules.

  1. Mycetoma caused by Nocardia madurae.

    PubMed Central

    Saksun, J. M.; Kane, J.; Schachter, R. K.

    1978-01-01

    Actinomycotic mycetoma was diagnosed in a woman from Jamaica living in Ontario. This is the first case reported in Canada in which the infection was caused by Nocardia madurae. Despite oral therapy with trimethoprim-sulfamethoxazole, local excision of newly appearing nodules was required periodically for clinical improvement. Laboratory procedures were modified to aid in identification of pathogenic actinomycetes. Images FIG. 1 FIG. 2 FIG. 3 FIG. 4 FIG. 5 FIG. 6 PMID:737642

  2. A unique challenge: Emergency egress and life support equipment at KSC

    NASA Technical Reports Server (NTRS)

    Waddell, H. M., Jr.

    1975-01-01

    As a result of the investigation following the January 1967 fire, which took the lives of three astronauts, materials were developed, flight hardware was modified, and test procedures were rewritten in order to establish the framework within which a more effective rescue concept could be developed. Topics discussed include breathing units, improved life support equipment, miniresuscitators, and hazardous tasks during space shuttle launch and landing operations.

  3. Coherent Raman Imaging of Live Muscle Sarcomeres Assisted by SFG Microscopy.

    PubMed

    Kim, Hyunmin; Kim, Do-Young; Joo, Kyung-Il; Kim, Jung-Hye; Jeong, Soon Moon; Lee, Eun Seong; Hahm, Jeong-Hoon; Kim, Kyuhyung; Moon, Dae Woon

    2017-08-23

    In this study, we used spectrally focused coherent anti-Stokes Raman scattering (spCARS) microscopy assisted by sum-frequency generation (SFG) to monitor the variations in the structural morphology and molecular vibrations of a live muscle of Caenorhabditis elegans. The subunits of the muscle sarcomeres, such as the M-line, myosin, dense body, and α-actinin, were alternatively observed using spCARS microscopy for different sample orientations, with the guidance of a myosin positional marker captured by SFG microscopy. Interestingly enough, the beam polarization dependence of the spCARS contrasts for two parallel subunits (dense body and myosin) showed a ~90° phase difference. The chemically sensitive spCARS spectra induced by the time-varying overlap of two pulses allowed (after a robust subtraction of the non-resonant background using a modified Kramers-Krönig transformation method) high-fidelity detection of various genetically modified muscle sarcomeres tuned to the C-H vibration (2800-3100 cm -1 ). Conversely, SFG image mapping assisted by phase-retrieved spCARS spectra also facilitated label-free monitoring of the changes in the muscle content of C. elegans that are associated with aging, based on the hypothesis that the C-H vibrational modes could serve as qualitative chemical markers sensitive to the amount and/or structural modulation of the muscle.

  4. Association between Sedentary Behaviour and Physical, Cognitive, and Psychosocial Status among Older Adults in Assisted Living.

    PubMed

    Leung, Pet-Ming; Ejupi, Andreas; van Schooten, Kimberley S; Aziz, Omar; Feldman, Fabio; Mackey, Dawn C; Ashe, Maureen C; Robinovitch, Stephen N

    2017-01-01

    Identification of the factors that influence sedentary behaviour in older adults is important for the design of appropriate intervention strategies. In this study, we determined the prevalence of sedentary behaviour and its association with physical, cognitive, and psychosocial status among older adults residing in Assisted Living (AL). Participants ( n = 114, mean age = 86.7) from AL sites in British Columbia wore waist-mounted activity monitors for 7 consecutive days, after being assessed with the Timed Up and Go (TUG), Montreal Cognitive Assessment (MoCA), Short Geriatric Depression Scale (GDS), and Modified Fall Efficacy Scale (MFES). On average, participants spent 87% of their waking hours in sedentary behaviour, which accumulated in 52 bouts per day with each bout lasting an average of 13 minutes. Increased sedentary behaviour associated significantly with scores on the TUG ( r = 0.373, p < 0.001) and MFES ( r = -0.261, p = 0.005), but not with the MoCA or GDS. Sedentary behaviour also associated with male gender, use of mobility aid, and multiple regression with increased age. We found that sedentary behaviour among older adults in AL associated with TUG scores and falls-related self-efficacy, which are modifiable targets for interventions to decrease sedentary behaviour in this population.

  5. Long-term viremia and fecal shedding in pups after modified-live canine parvovirus vaccination.

    PubMed

    Decaro, Nicola; Crescenzo, Giuseppe; Desario, Costantina; Cavalli, Alessandra; Losurdo, Michele; Colaianni, Maria Loredana; Ventrella, Gianpiero; Rizzi, Stefania; Aulicino, Stefano; Lucente, Maria Stella; Buonavoglia, Canio

    2014-06-24

    Canine parvovirus (CPV) modified live virus vaccines are able to infect vaccinated dogs replicating in the bloodstream and enteric mucosa. However, the exact duration and extent of CPV vaccine-induced viremia and fecal shedding are not known. With the aim to fill this gap, 26 dogs were administered two commercial vaccines containing a CPV-2 or CPV-2b strain and monitored for 28 days after vaccination. By using real-time PCR, vaccine-induced viremia and shedding were found to be long lasting for both vaccinal strains. Vaccinal CPV-2b shedding was detected for a shorter period than CPV-2 (12 against 19 mean days) but with greater viral loads, whereas viremia occurred for a longer period (22 against 19 mean days) and with higher titers for CPV-2b. Seroconversion appeared as early as 7 and 14 days post-vaccination for CPV-2b and CPV-2 vaccines, respectively. With no vaccine there was any diagnostic interference using in-clinic or hemagglutination test, since positive results were obtained only by fecal real-time PCR testing. The present study adds new insights into the CPV vaccine persistence in the organism and possible interference with diagnostic tests. Copyright © 2014 Elsevier Ltd. All rights reserved.

  6. Relationship between poverty and health among adolescents.

    PubMed

    Abernathy, Thomas J; Webster, Greg; Vermeulen, Marian

    2002-01-01

    Although the effect of low socioeconomic status on the health of adolescents has been documented, the mechanism by which this occurs is not well understood. Furthermore, improving adolescent health through public health policy typically requires the presence of one or more modifiable risk factors which can be targeted for intervention. In spite of the well-documented negative associations between poverty and health, few modifiable risk factors have been identified. This study used the Evans-Stoddart Model of Health and Well-Being as a framework to examine data on 1,759 adolescents, aged 12 to 19, collected as part of the 1994 National Population Health Survey. Results not only confirm the relationship between income and health, but suggest how the pathway operates through the social environment, lifestyle differences, access to health care, and a reduced sense of self-esteem and self-mastery. Bivariate and multivariate analyses found positive associations between physical activity levels and self-esteem and mastery. We interpret these findings as preliminary evidence that it might be possible to buffer the impact of poverty on health through policies which increase physical activity levels among those living in poverty. Such policies could also include a secondary goal of increasing the activity levels among inactive adolescents who are not living in poverty, as they will derive benefits from this increase, both psychologically and physiologically.

  7. Commercial porcine reproductive and respiratory syndrome virus (PRRSV)-2 modified live virus vaccine against heterologous single and dual Korean PRRSV-1 and PRRSV-2 challenge.

    PubMed

    Jeong, Jiwoon; Kim, Seeun; Park, Changhoon; Park, Kee Hwan; Kang, Ikjae; Park, Su-Jin; Chae, Chanhee

    2018-04-28

    This study evaluated porcine reproductive and respiratory syndrome virus (PRRSV)-2 modified live virus (MLV) vaccine against heterologous single and dual challenge of Korean PRRSV-1 and PRRSV-2. Pigs were administered PRRSV-2 MLV vaccine intramuscularly at 21 days of age and inoculated intranasally with both genotypes at 56 days of age. Vaccination of pigs with PRRSV-2 MLV vaccine resulted in reduction of viral loads of both PRRSV-1 and PRRSV-2 after heterologous single and dual challenge with PRRSV-1 and PRRSV-2. In addition, pigs vaccinated with PRRSV-2 MLV vaccine exhibited higher frequencies of PRRSV-1 and PRRSV-2 specific interferon-γ secreting cells (IFN-γ-SC) and showed a significant reduction in lung lesions and PRRSV nucleic acid within the lung lesions after single and dual challenge compared with unvaccinated challenged pigs. Taken together these results demonstrated that vaccination of pigs with PRRSV-2 is efficacious in protecting growing pigs from respiratory disease against heterologous single and dual PRRSV-1 and PRRSV-2 challenge. © British Veterinary Association (unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

  8. Improvement of Cell Survival During Human Pluripotent Stem Cell Definitive Endoderm Differentiation

    PubMed Central

    Wang, Han; Luo, Xie; Yao, Li; Lehman, Donna M.

    2015-01-01

    Definitive endoderm (DE) is a vital precursor for internal organs such as liver and pancreas. Efficient protocol to differentiate human embryonic stem cells (hESCs) or induced pluripotent stem cells (iPSCs) to DE is essential for regenerative medicine and for modeling diseases; yet, poor cell survival during DE differentiation remains unsolved. In this study, our use of B27 supplement in modified differentiation protocols has led to a substantial improvement. We used an SOX17-enhanced green fluorescent protein (eGFP) reporter hESC line to compare and modify established DE differentiation protocols. Both total live cell numbers and the percentages of eGFP-positive cells were used to assess differentiation efficiency. Among tested protocols, three modified protocols with serum-free B27 supplement were developed to generate a high number of DE cells. Massive cell death was avoided during DE differentiation and the percentage of DE cells remained high. When the resulting DE cells were further differentiated toward the pancreatic lineage, the expression of pancreatic-specific markers was significantly increased. Similar high DE differentiation efficiency was observed in H1 hESCs and iPSCs through the modified protocols. In B27 components, bovine serum albumin was found to facilitate DE differentiation and cell survival. Using our modified DE differentiation protocols, satisfactory quantities of quality DE can be produced as primary material for further endoderm lineage differentiation. PMID:26132288

  9. Effects of modified constraint-induced movement therapy with trunk restraint in early stroke patients: A single-blinded, randomized, controlled, pilot trial.

    PubMed

    Bang, Dae-Hyouk; Shin, Won-Seob; Choi, Ho-Suk

    2018-01-01

    Reducing compensatory strategies during repetitive upper-limb training may be helpful in relearning motor skills. To explore the effects of modified constraint-induced movement therapy (mCIMT), additionally modified by adding trunk restraint (TR), on upper-limb function and activities of daily living (ADLs) in early post-stroke patients. Twenty-four participants with early stroke were randomly assigned to mCIMT combined with TR (mCIMT + TR) or mCIMT alone. Each group underwent twenty sessions (1 h/d, 5 d/wk for 4 weeks). Patients were assessed with the action research arm test (ARAT), the Fugl-Meyer Assessment-Upper extremity (FMA-UE), the Modified Barthel index (MBI), the Maximal elbow extension angle during reaching (MEEAR), and Motor Activity Logs (MAL-AOU and MAL-QOM). The mCIMT + TR group exhibited greater improvement in the ARAT, FMA-UE, MBI, MEEAR, and MAL-AOU, and MAL-QOM than the mCIMT group. Statistical analyses showed significant differences in ARAT (P = 0.003), FMA-UE (P = 0.042), MBI (P = 0.001), MEEAR (P = 0.002), and MAL-AOU (P = 0.005) between the groups. Modified CIMT combined with TR may be more effective than mCIMT alone in improving upper-limb function and ADLs in patients with early stroke.

  10. Modifiable Lifestyle Behaviors Are Associated With Metabolic Syndrome in a Taiwanese Population.

    PubMed

    Lin, Kuei-Man; Chiou, Jeng-Yuan; Ko, Shu-Hua; Tan, Jung-Ying; Huang, Chien-Ning; Liao, Wen-Chun

    2015-11-01

    To explore associations between metabolic syndrome and modifiable lifestyle behaviors among the adult population in Taiwan. This cross-sectional study analyzed data from a nationally representative sample that participated in the 2005-2008 Nutrition and Health Survey in Taiwan. The sample (2,337 participants older than 19 years) provided data on demographic characteristics, modifiable lifestyle behaviors, anthropometric measurements, and blood chemistry panel. These data were analyzed by descriptive statistics, univariate logistic regression, and multivariate logistic regression to determine factors associated with metabolic syndrome. Metabolic syndrome had a prevalence of 25.2%, and this prevalence increased with age. In univariate regression analysis, metabolic syndrome was associated with age, living with family members, educational level, and modifiable lifestyle behaviors (smoking, drinking, betel quid chewing, and physical activity). Individuals with a smoking history and currently chewing betel quid had the highest risk for metabolic syndrome. The risk for metabolic syndrome might be reduced by public health campaigns to encourage people to quit smoking cigarettes and chewing betel quid. Implementing more modifiable lifestyle behaviors in daily life will decrease metabolic syndrome in Taiwan. Considering that betel quid chewing and tobacco smoking interact to adversely affect metabolic syndrome risk, public health campaigns against both behaviors seem to be a cost-effective and efficient health promotion strategy to reduce the prevalence rate of metabolic syndrome. © 2015 Sigma Theta Tau International.

  11. Gone with the Wind: Conceiving of Moral Responsibility in the Case of GMO Contamination.

    PubMed

    Robaey, Zoë

    2016-06-01

    Genetically modified organisms are a technology now used with increasing frequency in agriculture. Genetically modified seeds have the special characteristic of being living artefacts that can reproduce and spread; thus it is difficult to control where they end up. In addition, genetically modified seeds may also bring about uncertainties for environmental and human health. Where they will go and what effect they will have is therefore very hard to predict: this creates a puzzle for regulators. In this paper, I use the problem of contamination to complicate my ascription of forward-looking moral responsibility to owners of genetically modified organisms. Indeed, how can owners act responsibly if they cannot know that contamination has occurred? Also, because contamination creates new and unintended ownership, it challenges the ascription of forward-looking moral responsibility based on ownership. From a broader perspective, the question this paper aims to answer is as follows: how can we ascribe forward-looking moral responsibility when the effects of the technologies in question are difficult to know or unknown? To solve this problem, I look at the epistemic conditions for moral responsibility and connect them to the normative notion of the social experiment. Indeed, examining conditions for morally responsible experimentation helps to define a range of actions and to establish the related epistemic virtues that owners should develop in order to act responsibly where genetically modified organisms are concerned.

  12. Living well: an intervention to improve self-management of medical illness for individuals with serious mental illness.

    PubMed

    Goldberg, Richard W; Dickerson, Faith; Lucksted, Alicia; Brown, Clayton H; Weber, Elyssa; Tenhula, Wendy N; Kreyenbuhl, Julie; Dixon, Lisa B

    2013-01-01

    Individuals with serious mental illness have elevated rates of comorbid chronic general medical conditions and may benefit from interventions designed to support illness self-management. This study examined the effectiveness of a modified version of the Chronic Disease Self-Management Program called Living Well for individuals with serious mental illness. A total of 63 mental health consumers with serious mental illness and at least one concurrent chronic general medical condition were randomly assigned to receive the 13-session peer-cofacilitated Living Well intervention or usual care. Participants were evaluated on attitudinal, behavioral, and functional outcomes at baseline, at the end of the intervention, and at a two-month follow-up. Living Well participants showed significant postintervention improvements across a range of attitudinal (self-efficacy and patient activation), behavioral (illness self-management techniques), and functional (physical and emotional well-being and general health functioning) outcomes. Although attenuation of effect was observed for most outcomes at two months postintervention, evidence was found of continued improvement in general self-management behaviors (use of action planning, brainstorming, and problem-solving). Continued advantage was found for the Living Well group in other areas, such as health-related locus of control and reports of healthy eating and physical activity. Receipt of Living Well was associated with a notable decrease in use of the emergency room for medical care, although the between-group difference was not statistically significant. Living Well shows promise in helping mental health consumers more effectively manage chronic general medical conditions and experience improved functioning and well-being.

  13. Evaluation of serum immunoglobulins among individuals living near six Superfund sites.

    PubMed

    Williamson, Dhelia M; White, Mary C; Poole, Charles; Kleinbaum, David; Vogt, Robert; North, Kari

    2006-07-01

    Residents living in communities near Superfund sites have expressed concern that releases from these facilities affect their health, including adverse effects on their immune systems. We used data from six cross-sectional studies to evaluate whether people who live near several Superfund sites are more likely to have individual immunoglobulin test results (IgA, IgG, and IgM) below or above the reference range than those who live in comparison areas with no Superfund site. Study participants consisted of target-area residents who lived close to a Superfund site and comparison-area residents who were not located near any Superfund or hazardous waste sites. A consistent modeling strategy was used across studies to assess the magnitude of the relationship between area of residence and immunoglobulin test results, adjusting for potential confounders and effect modifiers. In all study areas, the results suggest that people who live near a Superfund site may have been more likely to have IgA test results above the reference range than comparison areas residents regardless of modeling strategy employed. The effect measures were larger for residents who lived in communities near military bases with groundwater contamination. For all analyses the wide confidence intervals reflect uncertainty in the magnitude of these effects. To adequately address the question of whether the immune system is affected by low-level exposures to hazardous substances, we recommend that more functional immunotoxicity tests be conducted in human populations where individual exposure information is available or when it can be reasonably estimated from environmental exposure measurements.

  14. The double-edged sword: How evolution can make or break a live-attenuated virus vaccine

    PubMed Central

    Hanley, Kathryn A.

    2012-01-01

    Even students who reject evolution are often willing to consider cases in which evolutionary biology contributes to, or undermines, biomedical interventions. Moreover the intersection of evolutionary biology and biomedicine is fascinating in its own right. This review offers an overview of the ways in which evolution has impacted the design and deployment of live-attenuated virus vaccines, with subsections that may be useful as lecture material or as the basis for case studies in classes at a variety of levels. Live- attenuated virus vaccines have been modified in ways that restrain their replication in a host, so that infection (vaccination) produces immunity but not disease. Applied evolution, in the form of serial passage in novel host cells, is a “classical” method to generate live-attenuated viruses. However many live-attenuated vaccines exhibit reversion to virulence through back-mutation of attenuating mutations, compensatory mutations elsewhere in the genome, recombination or reassortment, or changes in quasispecies diversity. Additionally the combination of multiple live-attenuated strains may result in competition or facilitation between individual vaccine viruses, resulting in undesirable increases in virulence or decreases in immunogenicity. Genetic engineering informed by evolutionary thinking has led to a number of novel approaches to generate live-attenuated virus vaccines that contain substantial safeguards against reversion to virulence and that ameliorate interference among multiple vaccine strains. Finally, vaccines have the potential to shape the evolution of their wild type counterparts in counter-productive ways; at the extreme vaccine-driven eradication of a virus may create an empty niche that promotes the emergence of new viral pathogens. PMID:22468165

  15. Recognition of Activities of Daily Living Based on Environmental Analyses Using Audio Fingerprinting Techniques: A Systematic Review

    PubMed Central

    Santos, Rui; Pombo, Nuno; Flórez-Revuelta, Francisco

    2018-01-01

    An increase in the accuracy of identification of Activities of Daily Living (ADL) is very important for different goals of Enhanced Living Environments and for Ambient Assisted Living (AAL) tasks. This increase may be achieved through identification of the surrounding environment. Although this is usually used to identify the location, ADL recognition can be improved with the identification of the sound in that particular environment. This paper reviews audio fingerprinting techniques that can be used with the acoustic data acquired from mobile devices. A comprehensive literature search was conducted in order to identify relevant English language works aimed at the identification of the environment of ADLs using data acquired with mobile devices, published between 2002 and 2017. In total, 40 studies were analyzed and selected from 115 citations. The results highlight several audio fingerprinting techniques, including Modified discrete cosine transform (MDCT), Mel-frequency cepstrum coefficients (MFCC), Principal Component Analysis (PCA), Fast Fourier Transform (FFT), Gaussian mixture models (GMM), likelihood estimation, logarithmic moduled complex lapped transform (LMCLT), support vector machine (SVM), constant Q transform (CQT), symmetric pairwise boosting (SPB), Philips robust hash (PRH), linear discriminant analysis (LDA) and discrete cosine transform (DCT). PMID:29315232

  16. Use of luciferase probes to measure ATP in living cells and animals.

    PubMed

    Morciano, Giampaolo; Sarti, Alba Clara; Marchi, Saverio; Missiroli, Sonia; Falzoni, Simonetta; Raffaghello, Lizzia; Pistoia, Vito; Giorgi, Carlotta; Di Virgilio, Francesco; Pinton, Paolo

    2017-08-01

    ATP, the energy exchange factor that connects anabolism and catabolism, is required for major reactions and processes that occur in living cells, such as muscle contraction, phosphorylation and active transport. ATP is also the key molecule in extracellular purinergic signaling mechanisms, with an established crucial role in inflammation and several additional disease conditions. Here, we describe detailed protocols to measure the ATP concentration in isolated living cells and animals using luminescence techniques based on targeted luciferase probes. In the presence of magnesium, oxygen and ATP, the protein luciferase catalyzes oxidation of the substrate luciferin, which is associated with light emission. Recombinantly expressed wild-type luciferase is exclusively cytosolic; however, adding specific targeting sequences can modify its cellular localization. Using this strategy, we have constructed luciferase chimeras targeted to the mitochondrial matrix and the outer surface of the plasma membrane. Here, we describe optimized protocols for monitoring ATP concentrations in the cytosol, mitochondrial matrix and pericellular space in living cells via an overall procedure that requires an average of 3 d. In addition, we present a detailed protocol for the in vivo detection of extracellular ATP in mice using luciferase-transfected reporter cells. This latter procedure may require up to 25 d to complete.

  17. Resilience amongst Older Colombians Living in Poverty: an Ecological Approach.

    PubMed

    Bennett, Kate M; Reyes-Rodriguez, Maria F; Altamar, Paula; Soulsby, Laura K

    2016-12-01

    Older Colombians face significant adversities: poverty, violence and displacement. However, there is evidence that Latinos are often resilient. We examine resilience in older Colombians living in poverty using an ecological framework that identifies three levels: individual; community; and societal. In this paper we examine data from 16 semi-structured interviews with older Colombians that explore resilience within the context of poverty. We analyze our data using three stages: (1) modified grounded theory; (2) assignment of resilience status; (3) identification of components of the ecological framework which contribute to resilience in these participants. The most striking feature is that some participants are able to adapt to their situation, demonstrating resilience, whilst others are not. Individual characteristics such as psychological and material resources contribute to resilience. At the community level, family, social support, participation and cohesion promote resilience. Finally, at the societal level, social and welfare services, finance, religion and social policy, are important factors. These different levels of resilience are co-dependent, and we illustrate how this is so. We suggest that older Colombians living in poverty often demonstrate resilience, but that more can be done to enhance their lives. This includes interventions at the individual and community levels alongside changes in social policy.

  18. Living Cell Microarrays: An Overview of Concepts

    PubMed Central

    Jonczyk, Rebecca; Kurth, Tracy; Lavrentieva, Antonina; Walter, Johanna-Gabriela; Scheper, Thomas; Stahl, Frank

    2016-01-01

    Living cell microarrays are a highly efficient cellular screening system. Due to the low number of cells required per spot, cell microarrays enable the use of primary and stem cells and provide resolution close to the single-cell level. Apart from a variety of conventional static designs, microfluidic microarray systems have also been established. An alternative format is a microarray consisting of three-dimensional cell constructs ranging from cell spheroids to cells encapsulated in hydrogel. These systems provide an in vivo-like microenvironment and are preferably used for the investigation of cellular physiology, cytotoxicity, and drug screening. Thus, many different high-tech microarray platforms are currently available. Disadvantages of many systems include their high cost, the requirement of specialized equipment for their manufacture, and the poor comparability of results between different platforms. In this article, we provide an overview of static, microfluidic, and 3D cell microarrays. In addition, we describe a simple method for the printing of living cell microarrays on modified microscope glass slides using standard DNA microarray equipment available in most laboratories. Applications in research and diagnostics are discussed, e.g., the selective and sensitive detection of biomarkers. Finally, we highlight current limitations and the future prospects of living cell microarrays. PMID:27600077

  19. Conducting participatory photography with children with disabilities: a literature review.

    PubMed

    Eisen, Isabel; Cunningham, Barbara Jane; Campbell, Wenonah

    2018-03-28

    This review summarized studies that used participatory photography with children with disabilities, including those with communication impairments, and described modifications made to the methodology to facilitate their participation in qualitative research. In the fall of 2016, we searched Psycinfo (OVID), ERIC, CINAHL and Web of Science to identify studies that used participatory photography with children with disabilities. The search was repeated in January 2018 to retrieve any new publications. The first author extracted data that described the characteristics of each study and the modifications used. Of the 258 articles identified, 19 met inclusion criteria. Participants ranged from 4-21 years old and had a variety of disabilities. Study topics included education, leisure activities and adulthood. Researchers modified participatory photography to enhance accessibility by: modifying cameras; providing individual training; teaching consent through role play; allowing children to direct adults to take photographs; including additional forms of media; using diaries and questionnaires; providing individual interviews with simplified questions; using multiple forms of communication; and modifying how photographs are shared. Participatory photography can be an effective method for studying the lived experiences of children with disabilities, particularly those with communication impairments. Methodological modifications can enhance the accessibility of this approach for this population. Implications for Rehabilitation Participatory photography may be an effective qualitative research method for learning about the perspectives and experiences of children with disabilities on a wide array of topics. There are many specific modifications that researchers can use to support the inclusion of children with disabilities in participatory photography research. The findings of studies that use participatory photography methodology may provide rehabilitation professionals with important insights into the lives of children with disabilities.

  20. Therapeutic Potential of a Drum and Dance Ceremony Based on the African Ngoma Tradition.

    PubMed

    Vinesett, Ava L; Price, Miurel; Wilson, Kenneth H

    2015-08-01

    Ngoma ceremonies are used throughout Central and South Africa to help people address "difficult issues," including medical illness. They are examples of ceremonies that use strong rhythms and dance for this purpose in indigenous cultures throughout the world. This study sought to modify an ngoma ceremony to make it appropriate for biomedical use and to determine its acceptance and potential for benefit for people living in the United States. The Congolese Zebola ceremony, an African healing practice, was modified to be religion-neutral and to involve only moderate exercise. Seventeen participants were recruited for the current study. Most participants were living with a chronic illness (n=15), and a few had no medical diagnoses (n=2). Participants spent 10 minutes in a focused activity, such as meditation, yoga, or prayer. They then danced to the Congolese rhythm Zebola for an hour and a half, with a rest every 20 minutes. Afterward, they indicated whether the experience was positive, neutral, or negative and wrote a narrative describing their experience and what they saw as strengths and weaknesses of the ceremony. They then participated in a focus group discussion. Data from the narrative and focus group discussion were coded, tabulated, and analyzed for themes. Sixteen of seventeen participants found the ceremony to be a positive experience. None found the form uncomfortably foreign or disturbing. Participants described diverse benefits, including increased exercise tolerance, stress reduction, feelings of group support, and beneficial spiritual experiences. Further study of the modified ngoma ceremony is warranted. Global health programs should consider the potential benefits of transferring technologies in both directions rather than only from technologically advanced countries to less technologically advanced ones.

  1. Therapeutic Potential of a Drum and Dance Ceremony Based on the African Ngoma Tradition

    PubMed Central

    Vinesett, Ava L.; Price, Miurel

    2015-01-01

    Abstract Objective: Ngoma ceremonies are used throughout Central and South Africa to help people address “difficult issues,” including medical illness. They are examples of ceremonies that use strong rhythms and dance for this purpose in indigenous cultures throughout the world. This study sought to modify an ngoma ceremony to make it appropriate for biomedical use and to determine its acceptance and potential for benefit for people living in the United States. Methods: The Congolese Zebola ceremony, an African healing practice, was modified to be religion-neutral and to involve only moderate exercise. Seventeen participants were recruited for the current study. Most participants were living with a chronic illness (n=15), and a few had no medical diagnoses (n=2). Participants spent 10 minutes in a focused activity, such as meditation, yoga, or prayer. They then danced to the Congolese rhythm Zebola for an hour and a half, with a rest every 20 minutes. Afterward, they indicated whether the experience was positive, neutral, or negative and wrote a narrative describing their experience and what they saw as strengths and weaknesses of the ceremony. They then participated in a focus group discussion. Data from the narrative and focus group discussion were coded, tabulated, and analyzed for themes. Results: Sixteen of seventeen participants found the ceremony to be a positive experience. None found the form uncomfortably foreign or disturbing. Participants described diverse benefits, including increased exercise tolerance, stress reduction, feelings of group support, and beneficial spiritual experiences. Conclusions: Further study of the modified ngoma ceremony is warranted. Global health programs should consider the potential benefits of transferring technologies in both directions rather than only from technologically advanced countries to less technologically advanced ones. PMID:26171652

  2. Living Independently: Exploring the Experiences of Visually Impaired People Living in Age-Related and Lifetime Housing Through Qualitative Synthesis.

    PubMed

    Rooney, Clíona; Hadjri, Karim; Faith, Verity; Rooney, Máirin; McAllister, Keith; Craig, Cathy

    2017-01-01

    The aim of this study is to gain a deeper understanding of the experiences of visually impaired older people living independently at home. As populations are aging globally, there is now an increase in the prevalence of visual impairment. That means for ongoing and future aging-in-place strategies that seek to enable older people to remain independent for longer, more attention needs to be given to the needs of those with visual impairment. As people develop visual impairment, they use adaptive strategies including modifying long-term homes or relocating to more suitable accommodation. In the United Kingdom, aging-in-place strategies include employing statutory lifetime home standards (LTHS) in the home or relocating to sheltered housing to live independently with support available if required. To get a better understanding of the needs of the visually impaired in the home, 12 interviews with six visually impaired occupants of LTHS homes and six from sheltered accommodation were analyzed separately using interpretative phenomenological analysis. Secondly, qualitative synthesis was used to further analyze themes generated from both samples before interview results were conceptualized in two superordinate concepts, namely, "negotiating priorities" and "understanding visual impairment." Participants from both groups had similar needs and were willing to compromise by living with some negative features. Those who coped well with moving utilized various resources. These findings will shed more understanding on providing good quality housing for those with visual impairment wanting to live either independently or within healthcare home environments.

  3. High prevalence of body dysmorphic disorder symptoms in patients seeking rhinoplasty.

    PubMed

    Picavet, Valerie A; Prokopakis, Emmanuel P; Gabriëls, Lutgardis; Jorissen, Mark; Hellings, Peter W

    2011-08-01

    Nasal aesthetic deformities may be associated with significant body image dissatisfaction. The only diagnostic category in the current list of psychiatric disorders that directly addresses these concerns is body dysmorphic disorder. This large-scale study determined the prevalence of body dysmorphic disorder and its symptoms in patients seeking rhinoplasty and evaluated the clinical profile of these patients. Two hundred twenty-six patients were given questionnaires including demographic characteristics, visual analogue scales for nasal shape, the Yale-Brown Obsessive Compulsive Scale modified for body dysmorphic disorder to assess severity of symptoms, a generic quality-of-life questionnaire, and the Derriford Appearance Scale 59, to assess appearance-related disruption of everyday living. Independent observers scored the nasal shape. Thirty-three percent of patients showed at least moderate symptoms of body dysmorphic disorder. Aesthetic goals (p < 0.001), revision rhinoplasty (p = 0.003), and psychiatric history (p = 0.031) were associated with more severe symptoms. There was no correlation between the objective and subjective scoring of the nasal shape. Yale-Brown scale modified for body dysmorphic disorder scores correlated inversely with the subjective nasal scoring (n = 210, p < 0.001), without relation to the objective deformity of the nose. Body dysmorphic disorder symptoms significantly reduced the generic quality of life (n = 160, p < 0.001) and led to significant appearance-related disruption of everyday living (n = 161, p < 0.001). The prevalence of moderate to severe body dysmorphic disorder symptoms in an aesthetic rhinoplasty population is high. Patients undergoing revision rhinoplasty and with psychiatric history are particularly at risk. Body dysmorphic disorder symptoms significantly reduce the quality of life and cause significant appearance-related disruption of everyday living. Risk, III.

  4. Probiotic E. coli Nissle 1917 biofilms on silicone substrates for bacterial interference against pathogen colonization.

    PubMed

    Chen, Quan; Zhu, Zhiling; Wang, Jun; Lopez, Analette I; Li, Siheng; Kumar, Amit; Yu, Fei; Chen, Haoqing; Cai, Chengzhi; Zhang, Lijuan

    2017-03-01

    Bacterial interference is an alternative strategy to fight against device-associated bacterial infections. Pursuing this strategy, a non-pathogenic bacterial biofilm is used as a live, protective barrier to fence off pathogen colonization. In this work, biofilms formed by probiotic Escherichia coli strain Nissle 1917 (EcN) are investigated for their potential for long-term bacterial interference against infections associated with silicone-based urinary catheters and indwelling catheters used in the digestive system, such as feeding tubes and voice prostheses. We have shown that EcN can form stable biofilms on silicone substrates, particularly those modified with a biphenyl mannoside derivative. These biofilms greatly reduced the colonization by pathogenic Enterococcus faecalis in Lysogeny broth (LB) for 11days. Bacterial interference is an alternative strategy to fight against device-associated bacterial infections. Pursuing this strategy, we use non-pathogenic bacteria to form a biofilm that serves as a live, protective barrier against pathogen colonization. Herein, we report the first use of preformed probiotic E. coli Nissle 1917 biofilms on the mannoside-presenting silicone substrates to prevent pathogen colonization. The biofilms serve as a live, protective barrier to fence off the pathogens, whereas current antimicrobial/antifouling coatings are subjected to gradual coverage by the biomass from the rapidly growing pathogens in a high-nutrient environment. It should be noted that E. coli Nissle 1917 is commercially available and has been used in many clinical trials. We also demonstrated that this probiotic strain performed significantly better than the non-commercial, genetically modified E. coli strain that we previously reported. Copyright © 2017 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  5. Liver Stiffness Measurements Using Acoustic Radiation Force Impulse in Recipients of Living-Donor and Deceased-Donor Orthotopic Liver Transplant.

    PubMed

    Haberal, Kemal Murat; Turnaoğlu, Hale; Özdemir, Adnan; Uslu, Nihal; Haberal Reyhan, Asuman Nihan; Moray, Gökhan; Haberal, Mehmet

    2017-08-24

    The aim of this study was to evaluate the diagnostic efficiency of the acoustic radiation force impulse (Siemens Medical Solutions, Erlangen, Germany) elastography in assessment of fibrosis in orthotopic liver transplant patients. We enrolled 28 orthotopic liver transplant patients (deceased and living donors), whose biopsy decision had been prospectively given clinically. Ten acoustic radiation force impulse elastographic measurements were applied before the biopsy or within 3 days after the biopsy by 2 radiologists. After the core tissue needle biopsy, specimens of all patients were analyzed according to the modified Ishak scoring system. Measurements of acoustic radiation force impulse elastography and pathology specimen results were compared. From 28 biopsies, fibrosis scores of 4 biopsies were evaluated as F0 (14.3%), 16 as F1 (57.1%), 4 as F2 (14.3%), and 4 as F3 (14.3%). Mean results of acoustic radiation force impulse measurements were calculated as 1.4 ± 0.07 in F0, 1.74 ± 0.57 in F1, 2.19 ± 0.7 in F2, and 2.18 ± 0.35 in F3. There were no significant correlations of mean acoustic radiation force impulse values between the F0 versus F1 (P = .956) and F0 versus F2 stages (P = .234). A statistically significant correlation of mean acoustic radiation force impulse values was found between the F0 and F3 fibrosis stages (P = .046). Acoustic radiation force impulse imaging is a promising screening test for detecting significant liver fibrosis (≥ F3 in modified Ishak) in living-donor or deceased-donor orthotopic liver transplant recipients.

  6. Date palm pollen allergoid: characterization of its chemical-physical and immunological properties.

    PubMed

    Mistrello, G; Harfi, H; Roncarolo, D; Kwaasi, A; Zanoni, D; Falagiani, P; Panzani, R

    2008-01-01

    Date palm (DP) pollen can cause allergic symptoms in people living in different countries. Specific immunotherapy with allergenic extracts by subcutaneous route is effective to cure allergic people. However, the risk of side effects has led to explore safer therapeutic modalities. The aim of our work was to evaluate IgE cross-reactivity between DP and autochthonous palm (European fan palm, EFP) pollen extracts, to chemically modify DP extract with potassium cyanate in order to obtain an allergoid, and to characterize it. By radioallergosorbent test inhibition, immunoblotting (IB) and skin prick test, in vitro and in vivo allergenic activities of native and modified DP extracts were compared. By SDS-PAGE and IB, we compared the protein profile and IgE-binding capacity of both native and modified DP, as well as of EFP extracts. By IB inhibition, IgE cross-reactivity of native DP and EFP extracts was evaluated. By ELISA, the capacity of modified DP-induced IgG to react with native DP extract was determined. Radioallergosorbent test inhibition, IB and skin prick test results demonstrated that modified DP was significantly less allergenic than native DP extract. The SDS-PAGE profile showed that potassium cyanate treatment of DP extract did not alter the molecular weight of its components. In addition, no difference was observed between native DP and EFP extracts. Subsequent IB inhibition data evidenced the existence of a strong IgE cross-reactivity between native DP and EFP extracts. ELISA results indicated that the administration of modified DP in mice was able to induce specific IgG also recognizing native DP extract. Modified DP extract (allergoid) seems to be a good candidate for immunotherapy of patients affected by specific allergy. 2007 S. Karger AG, Basel

  7. Modifiable risk and protective factors for depressive symptoms in low-income African American mothers.

    PubMed

    Siefert, Kristine; Finlayson, Tracy L; Williams, David R; Delva, Jorge; Ismail, Amid I

    2007-01-01

    Low-income African American mothers of young children experience high rates of depression, but many of the risk factors that have been identified provide little direction for intervention. The authors examined modifiable risk and protective factors for probable depression (Center for Epidemiological Studies Depression Scale >or= 23) in 824 African American mothers living in the 39 poorest census tracts in Detroit. Household food insufficiency and deteriorated housing significantly increased the odds of likely depression, whereas availability of a loan in a crisis, help with childcare, and transportation were protective. However, more frequent experiences of everyday discrimination greatly increased the odds of elevated depressive symptoms. These findings support the need for interventions that operate across individual and societal levels to address the fundamental causes of poor mental health.

  8. Artificial Cell Therapy: New Strategies for the Therapeutic Delivery of Live Bacteria

    PubMed Central

    2005-01-01

    There has been rapid growth in research regarding the use of live bacterial cells for therapeutic purposes. The recognition that these cells can be genetically engineered to synthesize products that have therapeutic potential has generated considerable interest and excitement among clinicians and health professionals. It is expected that a wide range of disease modifying substrates such as enzymes, hormones, antibodies, vaccines, and other genetic products will be used successfully and will impact upon health care substantially. However, a major limitation in the use of these bacterial cells is the complexity of delivering them to the correct target tissues. Oral delivery of live cells, lyophilized cells, and immobilized cells has been attempted but with limited success. Primarily, this is because bacterial cells are incapable of surviving passage through the gastrointestinal tract. In many occasions, when given orally, these cells have been found to provoke immunogenic responses that are undesirable. Recent studies show that these problems can be overcome by delivering live bacterial cells, such as genetically engineered cells, using artificial cell microcapsules. This review summarizes recent advances in the therapeutic use of live bacterial cells for therapy, discusses the principles of using artificial cells for the oral delivery of bacterial cells, outlines methods for preparing suitable artificial cells for this purpose, addresses potentials and limitations for their application in therapy, and provides insight for the future direction of this emergent and highly prospective technology. PMID:15689638

  9. A mutualism without honeydew: what benefits for Melissotarsus emeryi ants and armored scale insects (Diaspididae)?

    PubMed

    Peeters, Christian; Foldi, Imre; Matile-Ferrero, Danièle; Fisher, Brian L

    2017-01-01

    Mutualisms between ants and sap-sucking insects generally involve clear benefits for both partners: the ants provide protection in exchange for honeydew. However, a single ant genus associates with armoured scale insects (Diaspididae) that do not excrete honeydew. We studied three colonies of Melissotarsus emeryi ants from two localities in Mozambique. Vast numbers of the diaspidid Morganella conspicua occupied galleries dug by the ants under the bark of living trees. Unlike free-living M. conspicua and other diaspidids, M. conspicua living with ants are known to lack shields, likely because they gain protection against enemies and desiccation. Nevertheless, we documented the occurrence of rare individuals with shields inside ant galleries, indicating that their glands continue to secrete wax and proteins as building material. This is likely to constitute a significant portion of the ants' diet, in addition to diaspidid exuviae and excretions from the Malpighian tubules. Indeed, Melissotarsus workers cannot walk outside the galleries due to modified middle legs, forcing them to obtain all nourishment within the tree. Melissotarsus founding queens, however, must locate a suitable host tree while flying, and acquire diaspidid crawlers. This mutualism involves ants that are highly specialised to chew through living wood, and diaspidids that can also live freely outside the bark. It is extremely widespread in Africa and Madagascar, recorded from 20 tree families, and harmful effects on plant hosts require rapid study.

  10. Knowledge, attitude and perception of second-hand smoke and factors promoting smoking in Malaysian adolescents.

    PubMed

    Abidin, N Zainol; Zulkifli, A; Abidin, E Zainal; Rasdi, I; Ismail, S N Syed; Rahman, A Abd; Hashim, Z; Semple, S

    2014-07-01

    To identify the relationship between knowledge, attitude and perception regarding environmental tobacco smoke (ETS) and smoking among Malaysian adolescents living in states with complete or partial smoke-free legislation (SFL). A total of 898 respondents aged 13-14 years were randomly selected from 21 secondary schools. The Malay version of the modified Global Youth Tobacco Survey questionnaire was used. Hierarchical logistic regression was performed in examining predictors of smoking attempt among adolescents. Participants exposed to ETS >5 h/day were more likely to have smoked compared to those exposed to ETS <1 h/day (OR 4.1, 95%CI 1.03-16.0). Living in a state with partial SFL was associated with a two-fold increase in self-reported smoking attempts (OR 1.95, 95%CI 1.10-3.43) compared to living in a state with complete SFL. Negative attitudes and perceptions towards smoking and ETS exposure were linked to lower smoking attempts in states with complete SFL. Adolescents with limited ETS exposure who lived in a state with complete SFL were less likely to attempt smoking compared to those exposed more regularly to ETS and living in a state with partial SFL. Preventing adolescents from becoming smokers is the key to reducing national prevalence rates in smoking. There is a need to implement comprehensive smoke-free legislation nationally across Malaysia.

  11. A mutualism without honeydew: what benefits for Melissotarsus emeryi ants and armored scale insects (Diaspididae)?

    PubMed Central

    Foldi, Imre; Matile-Ferrero, Danièle; Fisher, Brian L.

    2017-01-01

    Mutualisms between ants and sap-sucking insects generally involve clear benefits for both partners: the ants provide protection in exchange for honeydew. However, a single ant genus associates with armoured scale insects (Diaspididae) that do not excrete honeydew. We studied three colonies of Melissotarsus emeryi ants from two localities in Mozambique. Vast numbers of the diaspidid Morganella conspicua occupied galleries dug by the ants under the bark of living trees. Unlike free-living M. conspicua and other diaspidids, M. conspicua living with ants are known to lack shields, likely because they gain protection against enemies and desiccation. Nevertheless, we documented the occurrence of rare individuals with shields inside ant galleries, indicating that their glands continue to secrete wax and proteins as building material. This is likely to constitute a significant portion of the ants’ diet, in addition to diaspidid exuviae and excretions from the Malpighian tubules. Indeed, Melissotarsus workers cannot walk outside the galleries due to modified middle legs, forcing them to obtain all nourishment within the tree. Melissotarsus founding queens, however, must locate a suitable host tree while flying, and acquire diaspidid crawlers. This mutualism involves ants that are highly specialised to chew through living wood, and diaspidids that can also live freely outside the bark. It is extremely widespread in Africa and Madagascar, recorded from 20 tree families, and harmful effects on plant hosts require rapid study. PMID:28761787

  12. Effect of trauma on quality of life as mediated by mental distress and moderated by coping and social support among postconflict displaced Ethiopians.

    PubMed

    Araya, Mesfin; Chotai, Jayanti; Komproe, Ivan H; de Jong, Joop T V M

    2007-08-01

    An understanding of how quality of life is affected by severe trauma and mental distress may facilitate better intervention strategies for postconflict internally displaced persons, by identifying mediators, moderators, and independent risk factors. We investigate the pathways involved in this process and also study the moderating roles of coping strategies and perceived social support. A random sample of 1193 (62% women) internally displaced Ethiopian adults living in shelters in Addis Ababa were interviewed with instruments capturing the relevant concepts, including SCL-90-R and WHOQOL-BREF. Path analysis was employed to elaborate the mediating and moderating effects. Self-reported living conditions were also assessed. Mental distress increased and quality of life decreased with age. Mental distress mediated the effects of trauma in reducing the quality of life, and some trauma also reduced quality of life directly. These effects remained after adjusting for living conditions. Living conditions were related to quality of life also on their own. Coping strategies and perceived social support influenced mental distress and quality of life directly as well as indirectly by moderation, in part gender specific. Intervention strategies aimed at reducing mental distress, modifying coping strategies, and encouraging social support may turn out to be useful in increasing the overall quality of life in postconflict situations, and are worth considering as complements to strategies that improve the living conditions.

  13. To be or not to be alive: How recent discoveries challenge the traditional definitions of viruses and life.

    PubMed

    Forterre, Patrick

    2016-10-01

    Three major discoveries have recently profoundly modified our perception of the viral world: molecular ecologists have shown that viral particles are more abundant than cells in natural environments; structural biologists have shown that some viruses from the three domains of life, Bacteria, Eukarya and Archaea, are evolutionarily related, and microbiologists have discovered giant viruses that rival with cells in terms of size and gene content. I discuss here the scientific and philosophical impact of these discoveries on the debates over the definition, nature (living or not), and origin of viruses. I suggest that viruses have often been considered non-living, because they are traditionally assimilated to their virions. However, the term virus describes a biological process and should integrate all aspects of the viral reproduction cycle. It is especially important to focus on the intracellular part of this cycle, the virocell, when viral information is actively expressed and reproduced, allowing the emergence of new viral genes. The virocell concept theoretically removes roadblocks that prevent defining viruses as living organisms. However, defining a "living organism" remains challenging, as indicated by the case of organelles that evolved from intracellular bacteria. To bypass this problem, I suggest considering that all biological entities that actively participate in the process of life are living. Copyright © 2016. Published by Elsevier Ltd.

  14. Urban reconciliation ecology: the potential of living roofs and walls.

    PubMed

    Francis, Robert A; Lorimer, Jamie

    2011-06-01

    Reconciling human and non-human use of urban regions to support biological conservation represents a major challenge for the 21st century. The concept of reconciliation ecology, by which the anthropogenic environment may be modified to encourage non-human use and biodiversity preservation without compromising societal utilization, potentially represents an appropriate paradigm for urban conservation given the generally poor opportunities that exist for reserve establishment and ecological restoration in urban areas. Two habitat improvement techniques with great potential for reconciliation ecology in urban areas are the installation of living roofs and walls, which have been shown to support a range of taxa at local scales. This paper evaluates the reconciliation potential of living roofs and walls, in particular highlighting both ecological and societal limitations that need to be overcome for application at the landscape scale. We further consider that successful utilization of living roofs and walls for urban reconciliation ecology will rely heavily on the participation of urban citizens, and that a 'citizen science' model is needed to facilitate public participation and support and to create an evidence base to determine their effectiveness. Living roofs and walls are just one aspect of urban reconciliation ecology, but are particularly important 'bottom-up' techniques for improving urban biodiversity that can be performed directly by the citizenry. Copyright © 2011 Elsevier Ltd. All rights reserved.

  15. Stimulating Parenting Practices in Indigenous and Non-Indigenous Mexican Communities

    PubMed Central

    Ozer, Emily J.; Dow, William

    2017-01-01

    Parenting may be influenced by ethnicity; marginalization; education; and poverty. A critical but unexamined question is how these factors may interact to compromise or support parenting practices in ethnic minority communities. This analysis examined associations between mothers’ stimulating parenting practices and a range of child-level (age; sex; and cognitive and socio-emotional development); household-level (indigenous ethnicity; poverty; and parental education); and community-level (economic marginalization and majority indigenous population) variables among 1893 children ages 4–18 months in poor; rural communities in Mexico. We also explored modifiers of associations between living in an indigenous community and parenting. Key findings were that stimulating parenting was negatively associated with living in an indigenous community or family self-identification as indigenous (β = −4.25; SE (Standard Error) = 0.98; β = −1.58; SE = 0.83 respectively). However; living in an indigenous community was associated with significantly more stimulating parenting among indigenous families than living in a non-indigenous community (β = 2.96; SE = 1.25). Maternal education was positively associated with stimulating parenting only in indigenous communities; and household crowding was negatively associated with stimulating parenting only in non-indigenous communities. Mothers’ parenting practices were not associated with child sex; father’s residential status; education; or community marginalization. Our findings demonstrate that despite greater community marginalization; living in an indigenous community is protective for stimulating parenting practices of indigenous mothers. PMID:29295595

  16. Stimulating Parenting Practices in Indigenous and Non-Indigenous Mexican Communities.

    PubMed

    Knauer, Heather A; Ozer, Emily J; Dow, William; Fernald, Lia C H

    2017-12-25

    Parenting may be influenced by ethnicity; marginalization; education; and poverty. A critical but unexamined question is how these factors may interact to compromise or support parenting practices in ethnic minority communities. This analysis examined associations between mothers' stimulating parenting practices and a range of child-level (age; sex; and cognitive and socio-emotional development); household-level (indigenous ethnicity; poverty; and parental education); and community-level (economic marginalization and majority indigenous population) variables among 1893 children ages 4-18 months in poor; rural communities in Mexico. We also explored modifiers of associations between living in an indigenous community and parenting. Key findings were that stimulating parenting was negatively associated with living in an indigenous community or family self-identification as indigenous (β = -4.25; SE (Standard Error) = 0.98; β = -1.58; SE = 0.83 respectively). However; living in an indigenous community was associated with significantly more stimulating parenting among indigenous families than living in a non-indigenous community (β = 2.96; SE = 1.25). Maternal education was positively associated with stimulating parenting only in indigenous communities; and household crowding was negatively associated with stimulating parenting only in non-indigenous communities. Mothers' parenting practices were not associated with child sex; father's residential status; education; or community marginalization. Our findings demonstrate that despite greater community marginalization; living in an indigenous community is protective for stimulating parenting practices of indigenous mothers.

  17. Coated microneedle arrays for transcutaneous delivery of live virus vaccines

    PubMed Central

    Vrdoljak, Anto; McGrath, Marie G.; Carey, John B.; Draper, Simon J.; Hill, Adrian V.S.; O’Mahony, Conor; Crean, Abina M.; Moore, Anne C.

    2016-01-01

    Vaccines are sensitive biologics that require continuous refrigerated storage to maintain their viability. The vast majority of vaccines are also administered using needles and syringes. The need for cold chain storage and the significant logistics surrounding needle-and-syringe vaccination is constraining the success of immunization programs. Recombinant live viral vectors are a promising platform for the development of vaccines against a number of infectious diseases, however these viruses must retain infectivity to be effective. Microneedles offer an effective and painless method for delivery of vaccines directly into skin that in the future could provide solutions to current vaccination issues. Here we investigated methods of coating live recombinant adenovirus and modified vaccinia virus Ankara (MVA) vectors onto solid microneedle arrays. An effective spray-coating method, using conventional pharmaceutical processes, was developed, in tandem with suitable sugar-based formulations, which produces arrays with a unique coating of viable virus in a dry form around the shaft of each microneedle on the array. Administration of live virus-coated microneedle arrays successfully resulted in virus delivery, transcutaneous infection and induced an antibody or CD8+ T cell response in mice that was comparable to that obtained by needle-and-syringe intradermal immunization. To our knowledge, this is the first report of successful vaccination with recombinant live viral vectored vaccines coated on microneedle delivery devices. PMID:22245683

  18. Coated microneedle arrays for transcutaneous delivery of live virus vaccines.

    PubMed

    Vrdoljak, Anto; McGrath, Marie G; Carey, John B; Draper, Simon J; Hill, Adrian V S; O'Mahony, Conor; Crean, Abina M; Moore, Anne C

    2012-04-10

    Vaccines are sensitive biologics that require continuous refrigerated storage to maintain their viability. The vast majority of vaccines are also administered using needles and syringes. The need for cold chain storage and the significant logistics surrounding needle-and-syringe vaccination is constraining the success of immunization programs. Recombinant live viral vectors are a promising platform for the development of vaccines against a number of infectious diseases, however these viruses must retain infectivity to be effective. Microneedles offer an effective and painless method for delivery of vaccines directly into skin that in the future could provide solutions to current vaccination issues. Here we investigated methods of coating live recombinant adenovirus and modified vaccinia virus Ankara (MVA) vectors onto solid microneedle arrays. An effective spray-coating method, using conventional pharmaceutical processes, was developed, in tandem with suitable sugar-based formulations, which produces arrays with a unique coating of viable virus in a dry form around the shaft of each microneedle on the array. Administration of live virus-coated microneedle arrays successfully resulted in virus delivery, transcutaneous infection and induced an antibody or CD8(+) T cell response in mice that was comparable to that obtained by needle-and-syringe intradermal immunization. To our knowledge, this is the first report of successful vaccination with recombinant live viral vectored vaccines coated on microneedle delivery devices. Copyright © 2011 Elsevier B.V. All rights reserved.

  19. Does living density matter for nonfatal unintentional home injury in Asian urban settings? Evidence from Hong Kong.

    PubMed

    Chan, Emily Y Y; Kim, Jean H; Griffiths, Sian M; Lau, Joseph T F; Yu, Ignatius

    2009-11-01

    Injury is a major global disease burden for the twenty-first century. There are, however, few studies of unintentional household injury in Asian urban settings where living environments are characterized by extremely compact, high-living-density, multistory apartments. This study investigated the association between nonfatal unintentional household injuries with the resident's sociodemographic attributes and household characteristics in Hong Kong, the city with the world's highest population density. A cross-sectional retrospective recall study was conducted in May 2007 using a random telephone survey with a modified Chinese version of the World Health Organization Injury and Violence instrument. The study sample included 1,001 noninstitutionalized Cantonese-speaking Hong Kong residents of all ages, including foreign live-in domestic helpers. Multivariate regression was conducted to identify risk factors for nonfatal unintentional injuries in Hong Kong. Among a predominantly adult sample, household size and time spent at home were not associated with nonfatal unintentional household injuries in the general population in Hong Kong. The multivariate analyses indicated that female gender, owners of private homes, lower square footage of living space per person, and those with slip prevention devices in the bathroom were significantly associated with household injuries. Injured and noninjured groups were found to have adopted different injury prevention strategies toward household injuries. The results identified potential target groups for household injury prevention programs.

  20. Screening for Social Determinants of Health Among Children and Families Living in Poverty: A Guide for Clinicians.

    PubMed

    Chung, Esther K; Siegel, Benjamin S; Garg, Arvin; Conroy, Kathleen; Gross, Rachel S; Long, Dayna A; Lewis, Gena; Osman, Cynthia J; Jo Messito, Mary; Wade, Roy; Shonna Yin, H; Cox, Joanne; Fierman, Arthur H

    2016-05-01

    Approximately 20% of all children in the United States live in poverty, which exists in rural, urban, and suburban areas. Thus, all child health clinicians need to be familiar with the effects of poverty on health and to understand associated, preventable, and modifiable social factors that impact health. Social determinants of health are identifiable root causes of medical problems. For children living in poverty, social determinants of health for which clinicians may play a role include the following: child maltreatment, child care and education, family financial support, physical environment, family social support, intimate partner violence, maternal depression and family mental illness, household substance abuse, firearm exposure, and parental health literacy. Children, particularly those living in poverty, exposed to adverse childhood experiences are susceptible to toxic stress and a variety of child and adult health problems, including developmental delay, asthma and heart disease. Despite the detrimental effects of social determinants on health, few child health clinicians routinely address the unmet social and psychosocial factors impacting children and their families during routine primary care visits. Clinicians need tools to screen for social determinants of health and to be familiar with available local and national resources to address these issues. These guidelines provide an overview of social determinants of health impacting children living in poverty and provide clinicians with practical screening tools and resources. Copyright © 2016 Mosby, Inc. All rights reserved.

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