Perceived Social Ecological Factors Associated with Fruit and Vegetable Purchasing, Preparation, and Consumption among Young Adults

PubMed Central

Graham, Dan J.; Pelletier, Jennifer; Neumark-Sztainer, Dianne; Lust, Katherine; Laska, Melissa N.

2013-01-01

Most young adults do not consume recommended levels of fruits and vegetables (FV), and interventions to increase FV-related behaviors among this understudied population are needed. Therefore, it is important to identify correlates of FV intake among young adults to guide intervention development. This cross-sectional study utilized data from an online survey to identify factors related to young adults’ FV purchasing, preparation, and consumption, and to explore between-factor relationships using mediation analysis. In 2010, 1201 college students in Minnesota completed questionnaires assessing FV behaviors, as well as perceptions of FV-related individual, social, and environmental factors. Factor analysis identified questionnaire items assessing similar constructs. Seven factors were identified (personal barriers, FV knowledge, family, friends, neighborhood, access barriers, and campus) and evaluated for relationships with FV purchasing, preparation, and consumption using linear regression. Results revealed that perceived personal barriers (e.g., lacking cooking skills) were inversely related to all FV outcomes. Perception that family and friends eat healthfully and neighborhood access to FV were positively related to all outcomes. Individual-, social-, and environmental-level perceptions were related to purchasing, preparation, and consumption, and the effects of these factors were similar when accounting for mediated effects. Factors at all three levels and the ways in which these various factors operate together may be important to consider in future efforts to improve FV behaviors among young adults. PMID:23958116

Interleukin-Encoding Adenoviral Vectors as Genetic Adjuvant for Vaccination against Retroviral Infection

PubMed Central

Ohs, Inga; Windmann, Sonja; Wildner, Oliver; Dittmer, Ulf; Bayer, Wibke

2013-01-01

Interleukins (IL) are cytokines with stimulatory and modulatory functions in the immune system. In this study, we have chosen interleukins which are involved in the enhancement of TH2 responses and B cell functions to analyze their potential to improve a prophylactic adenovirus-based anti-retroviral vaccine with regard to antibody and virus-specific CD4+ T cell responses. Mice were vaccinated with an adenoviral vector which encodes and displays the Friend Virus (FV) surface envelope protein gp70 (Ad.pIXgp70) in combination with adenoviral vectors encoding the interleukins IL4, IL5, IL6, IL7 or IL23. Co-application of Ad.pIXgp70 with Ad.IL5, Ad.IL6 or Ad.IL23 resulted in improved protection with high control over FV-induced splenomegaly and reduced viral loads. Mice co-immunized with adenoviral vectors encoding IL5 or IL23 showed increased neutralizing antibody responses while mice co-immunized with Ad.IL6 or Ad.IL23 showed improved FV-specific CD4+ T cell responses compared to mice immunized with Ad.pIXgp70 alone. We show that the co-application of adenoviral vectors encoding specific interleukins is suitable to improve the vaccination efficacy of an anti-retroviral vaccine. Improved protection correlated with improved CD4+ T cell responses and especially with higher neutralizing antibody titers. The co-application of selected interleukin-encoding adenoviral vectors is a valuable tool for vaccination with regard to enhancement of antibody mediated immunity. PMID:24349306

First detection of Apis mellifera filamentous virus in Apis cerana cerana in China.

PubMed

Hou, Chunsheng; Li, Beibei; Luo, Yuexiong; Deng, Shuai; Diao, Qingyun

2016-07-01

Although many honey bee RNA viruses have been correlated with colony declines, little is known regarding the potential role of DNA viruses. Here, we examined seemingly healthy and crawling bee samples from China using PCR to identify whether Apis mellifera filamentous virus (AmFV) was present in A. cerana cerana. The highest AmFV infection percentage among Chinese provinces occurred in crawling bees from Gansu province (85.48%), and the lowest was in bees from Beijing (31.58%). A phylogenetic analysis showed that the Chinese isolate of AmFV exhibited a high genetic similarity with isolates from Belgium, Switzerland and USA. This is the first report of AmFV infections in Chinese A. cerana cerana populations. Copyright © 2016 Elsevier Inc. All rights reserved.

Erythroid progenitor cells (CFU-E*) from Friend virus-infected mice undergo VVFe suicide in vitro in the absence of added erythropoietin

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

Del Rizzo, D.F.; Axelrad, A.A.

The authors have investigated the effect of VVFe on the survival in suspension of erythropoietin (epo)-independent erythroid progenitor cells (CFU-E*) induced by Friend polycythemia virus (FV). Spleen cells from C3Hf/Bi mice previously infected with FV were exposed to carrier-free VVFe, and the survival of CFU-E* as a function of time in liquid medium was determined from the number of erythroid colonies that developed from these cells seeded in plasma cultures without added epo. The results showed that spleen CFU-E* were highly vulnerable to VVFe. Marrow CFU-E* behaved in a similar manner. The VVFe responsible for their suicide had been presentedmore » to the progenitor cells only during the 4-h period of incubation, after which they were washed and plated in excess nonradioactive iron. They therefore conclude that CFU-E* themselves, and not only their progeny, are capable of actively incorporating iron. Under the same conditions in the absence of added epo, the effect of VVFe on the survival of normal spleen or marrow CFU-E could not be assessed because two few normal CFU-E survived the incubation period. Normal bone marrow cells incubated in complete medium containing epo retained their capacity for erythrocytic colony formation, and CFU-E could then be shown to be vulnerable to VVFe. Thus, either the iron-incorporating system of normal CFU-E was inducible by epo, or else epo permitted survival of the CFU-E so that the activity of a constitutive iron-incorporating system could be recognized.« less

Differential pH-dependent cellular uptake pathways among foamy viruses elucidated using dual-colored fluorescent particles

PubMed Central

2012-01-01

Background It is thought that foamy viruses (FVs) enter host cells via endocytosis because all FV glycoproteins examined display pH-dependent fusion activities. Only the prototype FV (PFV) glycoprotein has also significant fusion activity at neutral pH, suggesting that its uptake mechanism may deviate from other FVs. To gain new insights into the uptake processes of FV in individual live host cells, we developed fluorescently labeled infectious FVs. Results N-terminal tagging of the FV envelope leader peptide domain with a fluorescent protein resulted in efficient incorporation of the fluorescently labeled glycoprotein into secreted virions without interfering with their infectivity. Double-tagged viruses consisting of an eGFP-tagged PFV capsid (Gag-eGFP) and mCherry-tagged Env (Ch-Env) from either PFV or macaque simian FV (SFVmac) were observed during early stages of the infection pathway. PFV Env, but not SFVmac Env, containing particles induced strong syncytia formation on target cells. Both virus types showed trafficking of double-tagged virions towards the cell center. Upon fusion and subsequent capsid release into the cytosol, accumulation of naked capsid proteins was observed within four hours in the perinuclear region, presumably representing the centrosomes. Interestingly, virions harboring fusion-defective glycoproteins still promoted virus attachment and uptake, but failed to show syncytia formation and perinuclear capsid accumulation. Biochemical and initial imaging analysis indicated that productive fusion events occur predominantly within 4–6 h after virus attachment. Non-fused or non-fusogenic viruses are rapidly cleared from the cells by putative lysosomal degradation. Quantitative monitoring of the fraction of individual viruses containing both Env and capsid signals as a function of time demonstrated that PFV virions fused within the first few minutes, whereas fusion of SFVmac virions was less pronounced and observed over the entire 90 minutes measured. Conclusions The characterized double-labeled FVs described here provide new mechanistic insights into FV early entry steps, demonstrating that productive viral fusion occurs early after target cell attachment and uptake. The analysis highlights apparent differences in the uptake pathways of individual FV species. Furthermore, the infectious double-labeled FVs promise to provide important tools for future detailed analyses on individual FV fusion events in real time using advanced imaging techniques. PMID:22935135

Evaluation of the immunomodulatory and antiviral effects of the cytokine combination IFN-α and IL-7 in the lymphocytic choriomeningitis virus and Friend retrovirus mouse infection models.

PubMed

Audigé, Annette; Hofer, Ursula; Dittmer, Ulf; van den Broek, Maries; Speck, Roberto F

2011-10-01

Existing therapies for chronic viral infections are still suboptimal or have considerable side effects, so new therapeutic strategies need to be developed. One option is to boost the host's immune response with cytokines. We have recently shown in an acute ex vivo HIV infection model that co-administration of interferon (IFN)-α and interleukin (IL)-7 allows us to combine the potent anti-HIV activity of IFN-α with the beneficial effects of IL-7 on T-cell survival and function. Here we evaluated the effect of combining IFN-α and IL-7 on viral replication in vivo in the chronic lymphocytic choriomeningitis virus (LCMV) and acute Friend retrovirus (FV) infection models. In the chronic LCMV model, cytokine treatment was started during the early replication phase (i.e., on day 7 post-infection [pi]). Under the experimental conditions used, exogenous IFN-α inhibited FV replication, but had no effect on viral replication in the LCMV model. There was no therapeutic benefit of IL-7 either alone or in combination with IFN-α in either of the two infection models. In the LCMV model, dose-dependent effects of the cytokine combination on T-cell phenotype/function were observed. It is possible that these effects would translate into antiviral activity in re-challenged mice. It is also possible that another type of IFN-α/β or induction of endogenous IFN-α/β alone or in combination with IL-7 would have antiviral activity in the LCMV model. Furthermore, we cannot exclude that some effect on viral titers would have been seen at later time points not investigated here (i.e., beyond day 34 pi). Finally, IFN-α/IL-7 may inhibit the replication of other viruses. Thus it might be worth testing these cytokines in other in vivo models of chronic viral infections.

The Apis mellifera Filamentous Virus Genome

PubMed Central

Gauthier, Laurent; Cornman, Scott; Hartmann, Ulrike; Cousserans, François; Evans, Jay D.; de Miranda, Joachim R.; Neumann, Peter

2015-01-01

A complete reference genome of the Apis mellifera Filamentous virus (AmFV) was determined using Illumina Hiseq sequencing. The AmFV genome is a double stranded DNA molecule of approximately 498,500 nucleotides with a GC content of 50.8%. It encompasses 247 non-overlapping open reading frames (ORFs), equally distributed on both strands, which cover 65% of the genome. While most of the ORFs lacked threshold sequence alignments to reference protein databases, twenty-eight were found to display significant homologies with proteins present in other large double stranded DNA viruses. Remarkably, 13 ORFs had strong similarity with typical baculovirus domains such as PIFs (per os infectivity factor genes: pif-1, pif-2, pif-3 and p74) and BRO (Baculovirus Repeated Open Reading Frame). The putative AmFV DNA polymerase is of type B, but is only distantly related to those of the baculoviruses. The ORFs encoding proteins involved in nucleotide metabolism had the highest percent identity to viral proteins in GenBank. Other notable features include the presence of several collagen-like, chitin-binding, kinesin and pacifastin domains. Due to the large size of the AmFV genome and the inconsistent affiliation with other large double stranded DNA virus families infecting invertebrates, AmFV may belong to a new virus family. PMID:26184284

The Apis mellifera Filamentous Virus Genome.

PubMed

Gauthier, Laurent; Cornman, Scott; Hartmann, Ulrike; Cousserans, François; Evans, Jay D; de Miranda, Joachim R; Neumann, Peter

2015-07-09

A complete reference genome of the Apis mellifera Filamentous virus (AmFV) was determined using Illumina Hiseq sequencing. The AmFV genome is a double stranded DNA molecule of approximately 498,500 nucleotides with a GC content of 50.8%. It encompasses 247 non-overlapping open reading frames (ORFs), equally distributed on both strands, which cover 65% of the genome. While most of the ORFs lacked threshold sequence alignments to reference protein databases, twenty-eight were found to display significant homologies with proteins present in other large double stranded DNA viruses. Remarkably, 13 ORFs had strong similarity with typical baculovirus domains such as PIFs (per os infectivity factor genes: pif-1, pif-2, pif-3 and p74) and BRO (Baculovirus Repeated Open Reading Frame). The putative AmFV DNA polymerase is of type B, but is only distantly related to those of the baculoviruses. The ORFs encoding proteins involved in nucleotide metabolism had the highest percent identity to viral proteins in GenBank. Other notable features include the presence of several collagen-like, chitin-binding, kinesin and pacifastin domains. Due to the large size of the AmFV genome and the inconsistent affiliation with other large double stranded DNA virus families infecting invertebrates, AmFV may belong to a new virus family.

Enhanced transport of plant-produced rabies single-chain antibody-RVG peptide fusion protein across an in cellulo blood-brain barrier device.

PubMed

Phoolcharoen, Waranyoo; Prehaud, Christophe; van Dolleweerd, Craig J; Both, Leonard; da Costa, Anaelle; Lafon, Monique; Ma, Julian K-C

2017-10-01

The biomedical applications of antibody engineering are developing rapidly and have been expanded to plant expression platforms. In this study, we have generated a novel antibody molecule in planta for targeted delivery across the blood-brain barrier (BBB). Rabies virus (RABV) is a neurotropic virus for which there is no effective treatment after entry into the central nervous system. This study investigated the use of a RABV glycoprotein peptide sequence to assist delivery of a rabies neutralizing single-chain antibody (ScFv) across an in cellulo model of human BBB. The 29 amino acid rabies virus peptide (RVG) recognizes the nicotinic acetylcholine receptor (nAchR) at neuromuscular junctions and the BBB. ScFv and ScFv-RVG fusion proteins were produced in Nicotiana benthamiana by transient expression. Both molecules were successfully expressed and purified, but the ScFv expression level was significantly higher than that of ScFv-RVG fusion. Both ScFv and ScFv-RVG fusion molecules had potent neutralization activity against RABVin cellulo. The ScFv-RVG fusion demonstrated increased binding to nAchR and entry into neuronal cells, compared to ScFv alone. Additionally, a human brain endothelial cell line BBB model was used to demonstrate that plant-produced ScFv-RVG P fusion could translocate across the cells. This study indicates that the plant-produced ScFv-RVG P fusion protein was able to cross the in celluloBBB and neutralize RABV. © 2017 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

B cells and TCR avidity determine distinct functions of CD4+ T cells in retroviral infection1

PubMed Central

Ploquin, Mickaël J-Y; Eksmond, Urszula; Kassiotis, George

2011-01-01

The T-cell-dependent B-cell response relies on cognate interaction between B cells and CD4+ Th cells. However, the consequences of this interaction for CD4+ T cells are not entirely known. B cells generally promote CD4+ T-cell responses to pathogens, albeit to a variable degree. In contrast, CD4+ T-cell responses to self or tumor antigens are often suppressed by B cells. Here we demonstrated that interaction with B cells dramatically inhibited the function of virus-specific CD4+ T cells in retroviral infection. We have used Friend virus (FV) infection of mice as a model for retroviral infection, in which the behavior of virus-specific CD4+ T cells was monitored according to their TCR avidity. We report that avidity for antigen and interaction with B cells determine distinct aspects of the primary CD4+ T-cell response to FV infection. Virus-specific CD4+ T cells followed exclusive Th1 and T follicular helper (Tfh) differentiation. High avidity for antigen facilitated expansion during priming and enhanced the capacity for IFN-γ and IL-21 production. In contrast, Tfh differentiation was not affected by avidity for antigen. By reducing or preventing B-cell interaction we found that B cells promoted Tfh differentiation, induced programmed death 1 (PD-1) expression and inhibited IFN-γ production by virus-specific CD4+ T cells. Ultimately, B cells protected hosts from CD4+ T-cell-mediated immune pathology, at the detriment of CD4+ T-cell-mediated protective immunity. Our results suggest that B-cell presentation of vaccine antigens could be manipulated to direct the appropriate CD4+ T-cell response. PMID:21841129

Perceived social-ecological factors associated with fruit and vegetable purchasing, preparation, and consumption among young adults.

PubMed

Graham, Dan J; Pelletier, Jennifer E; Neumark-Sztainer, Dianne; Lust, Katherine; Laska, Melissa N

2013-10-01

Most young adults do not consume recommended levels of fruits and vegetables (F/V), and interventions to increase F/V-related behaviors among this understudied population are needed. Therefore, it is important to identify correlates of F/V intake among young adults to guide intervention development. This cross-sectional study used data from an online survey to identify factors related to young adults' F/V purchasing, preparation, and consumption, and to explore between-factor relationships using mediation analysis. In 2010, 1,201 college students in Minnesota completed questionnaires assessing F/V behaviors as well as perceptions of F/V-related individual, social, and environmental factors. Factor analysis identified questionnaire items assessing similar constructs. Seven factors were identified (personal barriers, F/V knowledge, family, friends, neighborhood, access barriers, and campus) and evaluated for relationships with F/V purchasing, preparation, and consumption using linear regression. Results revealed that perceived personal barriers (eg, lacking cooking skills) were inversely related to all F/V outcomes. Perception that family and friends eat healthfully and neighborhood access to F/V were positively related to all outcomes. Individual-, social-, and environment-level perceptions were related to purchasing, preparation, and consumption, and the effects of these factors were similar when accounting for mediated effects. Factors at all three levels and the ways in which these various factors operate together may be important to consider in future efforts to improve F/V behaviors among young adults. Copyright © 2013 Academy of Nutrition and Dietetics. Published by Elsevier Inc. All rights reserved.

Transfection of Fv-1 permissive and restrictive mouse cells with integrated DNA of murine leukemia viruses (host range restriction/Fv-1 gene/DNA transfection)

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

Hsu, I.C.; Yang, W.K.; Tennant, R.W.

1978-03-01

Whole-cell DNA preparations isolated from SC-1 cells chronically infected with N- or B-tropic murine leukemia viruses (MuLV) were tested for infectious activity in an Fv-1/sup n/ (NIH-3T3) and two Fv-1/sup b/ (C57BL/6 and SV-A31) cell cultures. Efficiency of transfection for all DNAs was better in the NIH-3T3 cells than in C57BL/6 or SV-A31 cells; and an (N-tropic MuLV)SC-1 cell DNA preparation was slightly more infectious than a (B-tropic MuLV)SC-1 cell DNA preparation in all three cell cultures, regardless of their Fv-1 geonotypes. Progeny viruses from the transfection showed N- or B-tropism corresponding to that of the parent viruses produced bymore » the infected SC-I cells that were used for the DNA preparation. DNA dose-response studies in NIH-3T3 cells revealed a one-hit mechanism for both the (B-tropic MuLV)SC-1 cell DNA and the (N-tropic MuLV)SC-1 cell DNA preparation. These results demonstrate that, in contrast to virion infection, transfection of N- and B-tropic MuLV with DNA preparations from chronically infected cells is not affected by the Fv-1 gene.« less

Engineering of a novel zipFv using leucine zipper motif against rabies virus glycoprotein G with improved protection potency in vivo.

PubMed

Xi, Hualong; Zhang, Kaixin; Yin, Yanchun; Gu, Tiejun; Sun, Qing; Li, Zhuang; Cheng, Yue; Jiang, Chunlai; Kong, Wei; Wu, Yongge

2017-06-01

Rabies is an acute zoonotic infectious disease with a high fatality rate but is preventable with vaccination and rabies immunoglobulin (RIG). The single-chain Fv fragment (scFv), a small engineered antigen-binding protein derived from antibody variable heavy (V H ) and light (V L ) chains connected by a peptide linker, can potentially be used to replace RIG. Here, we produced two peptides V H -JUN-HIS and V L -FOS-HA separately in Escherichia coli and assembled them to form zipFv successfully in vitro. The new zipFv utilizes FOS and JUN leucine zippers to form an antibody structure similar to the IgG counterpart with two free N-terminal ends of V H and V L . The zipFv protein showed notable improvement in binding ability and affinity over its corresponding scFv. The zipFv also demonstrated greater stability in serum and the same protective rate as RIG against challenge with a standard rabies virus (CVS-24) in mice. Our results indicated zipFv as a novel and efficient antibody form with enhanced neutralizing potency. Copyright © 2017. Published by Elsevier B.V.

Non-Simian Foamy Viruses: Molecular Virology, Tropism and Prevalence and Zoonotic/Interspecies Transmission

PubMed Central

Kehl, Timo; Tan, Juan; Materniak, Magdalena

2013-01-01

Within the field of retrovirus, our knowledge of foamy viruses (FV) is still limited. Their unique replication strategy and mechanism of viral persistency needs further research to gain understanding of the virus-host interactions, especially in the light of the recent findings suggesting their ancient origin and long co-evolution with their nonhuman hosts. Unquestionably, the most studied member is the primate/prototype foamy virus (PFV) which was originally isolated from a human (designated as human foamy virus, HFV), but later identified as chimpanzee origin; phylogenetic analysis clearly places it among other Old World primates. Additionally, the study of non-simian animal FVs can contribute to a deeper understanding of FV-host interactions and development of other animal models. The review aims at highlighting areas of special interest regarding the structure, biology, virus-host interactions and interspecies transmission potential of primate as well as non-primate foamy viruses for gaining new insights into FV biology. PMID:24064793

The Apis mellifera filamentous virus genome

USDA-ARS?s Scientific Manuscript database

A complete reference genome of the Apis mellifera Filamentous virus (AmFV) was determined using Illumina Hiseq sequencing. The AmFV genome is a double strand DNA molecule of approximately 498’500 nucleotides with a GC content of 50.8%. It encompasses 251 non overlapping open reading frames (ORFs), e...

Characterization of a novel single-stranded RNA virus, closely related to fusariviruses, infecting the plant pathogenic fungus Alternaria brassicicola.

PubMed

Zhong, Jie; Shang, Hong Hong; Zhu, Chuan Xia; Zhu, Jun Zi; Zhu, Hong Jian; Hu, Yan; Gao, Bi Da

2016-06-02

The alternaria blackspot of rapeseed is one of the most prominent diseases of rapeseed. It is caused by three species of the genus Alternaria: Alternaria brassicicola, Alternaria brassicae, and Alternaria raphanin. Here we report a novel positive-sense RNA virus from an A. brassicicola strain 817-14. The virus has a 6639 nucleotide (nt) long genome, excluding a poly (A)-tail, and was predicted to contain three putative open reading frames (ORF1, ORF2, and ORF3). The large ORF1 encoded a 174-kDa polyprotein (composed of 1522 amino acid residues) containing a conserved RNA-dependent RNA polymerase (RdRp) domain and a helicase domain. The other two smaller ORFs encoded polypeptides with unknown function. Homology search and phylogenetic analysis, based on the RdRp and helicase domains, suggest that this virus is related to and grouped with Sclerotinia sclerotiorum fusarivirus 1 (SsFV1), Rosellinia necatrix fusarivirus 1 (RnFV1), Fusarium graminearum virus-DK21 (FgV1), and Penicillium roqueforti RNA mycovirus 1 (PrRV1), all of which belong to a newly proposed family Fusariviridae. For this study, we designed the virus as "Alternaria brassicicola fusarivirus 1" (AbFV1). Virus elimination revealed that AbFV1 has no conspicuous impact on the biological properties of its host. Copyright © 2016. Published by Elsevier B.V.

Transmission dynamics of an insect-specific flavivirus in a naturally infected Culex pipiens laboratory colony and effects of co-infection on vector competence for West Nile virus

PubMed Central

Bolling, Bethany G.; Olea-Popelka, Francisco J.; Eisen, Lars; Moore, Chester G.; Blair, Carol D.

2012-01-01

We established a laboratory colony of Culex pipiens mosquitoes from eggs collected in Colorado and discovered that mosquitoes in the colony are naturally infected with Culex flavivirus (CxFV), an insect-specific flavivirus. In this study we examined transmission dynamics of CxFV and effects of persistent CxFV infection on vector competence for West Nile virus (WNV). We found that vertical transmission is the primary mechanism for persistence of CxFV in Cx. pipiens, with venereal transmission potentially playing a minor role. Vector competence experiments indicated possible early suppression of WNV replication by persistent CxFV infection in Cx. pipiens. This is the first description of insect-specific flavivirus transmission dynamics in a naturally infected mosquito colony and the observation of delayed dissemination of superinfecting WNV suggests that the presence of CxFV may impact the intensity of enzootic transmission of WNV and the risk of human exposure to this important pathogen. PMID:22425062

EXPERIMENTAL CHALLENGE STUDY OF FV3-LIKE RANAVIRUS INFECTION IN PREVIOUSLY FV3-LIKE RANAVIRUS INFECTED EASTERN BOX TURTLES (TERRAPENE CAROLINA CAROLINA) TO ASSESS INFECTION AND SURVIVAL.

PubMed

Hausmann, Jennifer C; Wack, Allison N; Allender, Matthew C; Cranfield, Mike R; Murphy, Kevin J; Barrett, Kevin; Romero, Jennell L; Wellehan, James F X; Blum, Stella A; Zink, M Christine; Bronson, Ellen

2015-12-01

The Maryland Zoo in Baltimore experienced an outbreak of Frog virus-3 (FV3)-like ranavirus during the summer of 2011, during which 14 of 27 (52%) of its captive eastern box turtles (Terrapene carolina carolina) survived. To assess survival, immunity, and viral shedding, an experimental challenge study was performed in which the surviving, previously infected turtles were reinfected with the outbreak strain of FV3-like ranavirus. Seven turtles were inoculated with virus intramuscularly and four control turtles received saline intramuscularly. The turtles were monitored for 8 wk with blood and oral swabs collected for quantitative polymerase chain reaction (qPCR). During that time, one of seven (14%) inoculated turtles and none of the controls (0%) died; there was no significant difference in survival. Clinical signs of the inoculated turtles, except for the turtle that died, were mild compared to the original outbreak. Quantitative PCR for FV3-like ranavirus on blood and oral swabs was positive for all inoculated turtles and negative for all controls. The turtle that died had intracytoplasmic inclusion bodies in multiple organs. Three inoculated and two control turtles were euthanized at the end of the study. No inclusion bodies were present in any of the organs. Quantitative PCR detected FV3-like ranavirus in the spleen of a control turtle, which suggested persistence of the virus. The surviving five turtles were qPCR-negative for FV3-like ranavirus from blood and oral swabs after brumation. Quantitative PCR for Terrapene herpesvirus 1 found no association between ranavirus infection and herpesvirus loads. In conclusion, previously infected eastern box turtles can be reinfected with the same strain of FV3-like ranavirus and show mild to no clinical signs but can shed the virus from the oral cavity.

Retargeting of adenovirus vectors through genetic fusion of a single-chain or single-domain antibody to capsid protein IX.

PubMed

Poulin, Kathy L; Lanthier, Robert M; Smith, Adam C; Christou, Carin; Risco Quiroz, Milagros; Powell, Karen L; O'Meara, Ryan W; Kothary, Rashmi; Lorimer, Ian A; Parks, Robin J

2010-10-01

Adenovirus (Ad) vectors are the most commonly used system for gene therapy applications, due in part to their ability to infect a wide array of cell types and tissues. However, many therapies would benefit from the ability to target the Ad vector only to specific cells, such as tumor cells for cancer gene therapy. In this study, we investigated the utility of capsid protein IX (pIX) as a platform for the presentation of single-chain variable-fragment antibodies (scFv) and single-domain antibodies (sdAb) for virus retargeting. We show that scFv can be displayed on the capsid through genetic fusion to native pIX but that these molecules fail to retarget the virus, due to improper folding of the scFv. Redirecting expression of the fusion protein to the endoplasmic reticulum (ER) results in correct folding of the scFv and allows it to recognize its epitope; however, ER-targeted pIX-scFv was incorporated into the Ad capsid at a very low level which was not sufficient to retarget virus infection. In contrast, a pIX-sdAb construct was efficiently incorporated into the Ad capsid and enhanced virus infection of cells expressing the targeted receptor. Taken together, our data indicate that pIX is an effective platform for presentation of large targeting polypeptides on the surface of the virus capsid, but the nature of the ligand can significantly affect its association with virions.

Isolation and Characterization of Viruses from the Kidneys of Rana pipiens with Renal Adenocarcinoma Before and After Passage in the Red Eft (Triturus viridescens)

PubMed Central

Clark, H. Fred; Brennan, James C.; Zeigel, Robert F.; Karzon, David T.

1968-01-01

Viruses were isolated from kidneys of normal and renal tumor-bearing Vermont Rana pipiens after subinoculation into red eft newts (Triturus viridescens). Organs of efts inoculated with viable cell suspensions from four of seven tumor-bearing kidneys yielded virus (LT-1, -2, -3, -4) when inoculated into TH-1 (Terrapene heart) cell culture. One tumor-bearing kidney also yielded virus (L-4) by direct inoculation into TH-1 cells. An additional isolate (L-5) was obtained from 1 of 52 normal Vermont frog kidneys inoculated directly into TH-1 cells. LT-1 was propagated with cytopathic effect (CPE) in each of 38 cell types tested, of fish, amphibian, reptilian, avian, and mammalian origin, at 23 or 30 C. LT-1 through LT-4, L-4 and L-5, and FV-1 through FV-3 each induced similar CPE in all cells tested. LT-2, however, induced CPE that progressed at a slower rate than that caused by the other isolates and produced smaller plaques (<0.8 mm) under starch gel overlay. Each of the viruses replicated to high titer in embryonated eggs incubated at 30 C. The viruses also grew in efts and adult newts, but not in bullfrog (Rana catesbeiana) tadpoles or adult leopard frogs. Tumor induction in adult leopard frogs inoculated with LT-1 was not demonstrated. Electron microscopic observations of LT-1 and LT-2 viruses revealed cytoplasmic particles, hexagonal in cross section, approximately 120 to 140 mμ in diameter, containing a dense nucleoid. LT-1 and LT-2 viruses were indistinguishable from FV-1 and Tipula iridescent virus. LT-1 was presumed to be a deoxyribonucleic acid virus on the basis of 5-bromodeoxyuridine inhibition. The isolates were ether-sensitive. On the basis of biological, physicochemical, and antigenic similarities, LT-1 through LT-4, L-4, L-5, FV-1 through FV-3, and isolates recently recovered from the bullfrog and the newt may represent strains of the same amphibian cytoplasmic virus. Images PMID:4972302

Transfection of Fv-1 permissive and restrictive mouse cells with integrated DNA of murine leukemia viruses

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

Hsu, I.C.; Yang, W.K.; Tennant, R.W.

1978-03-01

Whole-cell DNA preparations isolated from SC-1 cells chronically infected with N- or B-tropic murine leukemia viruses (MuLV) were tested for infectious activity in an Fv-I/sup n/ (NIH-3T3) and two Fv-I/sup b/ (C57BL/6 and SV-A31) cell cultures. Efficiency of transfection for all DNAs was better in the NIH-3T3 cells than in C57BL/6 or SV-A31 cells; and an (N-tropic MuLV)SC-1 cell DNA preparation was slightly more infectious than a (B-tropic MuLV)SC-1 cell DNA preparation in all three cell cultures, regardless of their Fv-1 genotypes. Progeny viruses from the transfection showed N- or B-tropism corresponding to that of the parent viruses produced bymore » the infected SC-1 cells that were used for the DNA preparation. DNA dose-response studies in NIH-3T3 cells revealed a one-hit mechanism for both the (B-tropic MuLV)SC-1 cell DNA and the (N-tropic MuLV)SC-1 cell DNA preparation. These results demonstrate that, in contrast to virion infection, transfection of N- and B-tropic MuLV with DNA preparations from chronically infected cells is not affected by the Fv-1 gene.« less

Large Animal Models for Foamy Virus Vector Gene Therapy

PubMed Central

Trobridge, Grant D.; Horn, Peter A.; Beard, Brian C.; Kiem, Hans-Peter

2012-01-01

Foamy virus (FV) vectors have shown great promise for hematopoietic stem cell (HSC) gene therapy. Their ability to efficiently deliver transgenes to multi-lineage long-term repopulating cells in large animal models suggests they will be effective for several human hematopoietic diseases. Here, we review FV vector studies in large animal models, including the use of FV vectors with the mutant O6-methylguanine-DNA methyltransferase, MGMTP140K to increase the number of genetically modified cells after transplantation. In these studies, FV vectors have mediated efficient gene transfer to polyclonal repopulating cells using short ex vivo transduction protocols designed to minimize the negative effects of ex vivo culture on stem cell engraftment. In this regard, FV vectors appear superior to gammaretroviral vectors, which require longer ex vivo culture to effect efficient transduction. FV vectors have also compared favorably with lentiviral vectors when directly compared in the dog model. FV vectors have corrected leukocyte adhesion deficiency and pyruvate kinase deficiency in the dog large animal model. FV vectors also appear safer than gammaretroviral vectors based on a reduced frequency of integrants near promoters and also near proto-oncogenes in canine repopulating cells. Together, these studies suggest that FV vectors should be highly effective for several human hematopoietic diseases, including those that will require relatively high percentages of gene-modified cells to achieve clinical benefit. PMID:23223198

Gladiolus plants transformed with single-chain variable fragment antibodies to Cucumber mosaic virus

USDA-ARS?s Scientific Manuscript database

Transgenic plants of Gladiolus ‘Peter Pears’ or ‘Jenny Lee’ were developed that contain single-chain variable fragments (scFv) to Cucumber mosaic virus (CMV) subgroup I or II. The CMV subgroup I heavy and light chain scFv fragments were placed under control of either the duplicated CaMV 35S or suga...

Transmission dynamics of an insect-specific flavivirus in a naturally infected Culex pipiens laboratory colony and effects of co-infection on vector competence for West Nile virus.

PubMed

Bolling, Bethany G; Olea-Popelka, Francisco J; Eisen, Lars; Moore, Chester G; Blair, Carol D

2012-06-05

We established a laboratory colony of Culex pipiens mosquitoes from eggs collected in Colorado and discovered that mosquitoes in the colony are naturally infected with Culex flavivirus (CxFV), an insect-specific flavivirus. In this study we examined transmission dynamics of CxFV and effects of persistent CxFV infection on vector competence for West Nile virus (WNV). We found that vertical transmission is the primary mechanism for persistence of CxFV in Cx. pipiens, with venereal transmission potentially playing a minor role. Vector competence experiments indicated possible early suppression of WNV replication by persistent CxFV infection in Cx. pipiens. This is the first description of insect-specific flavivirus transmission dynamics in a naturally infected mosquito colony and the observation of delayed dissemination of superinfecting WNV suggests that the presence of CxFV may impact the intensity of enzootic transmission of WNV and the risk of human exposure to this important pathogen. Copyright © 2012 Elsevier Inc. All rights reserved.

The association between motivation and fruit and vegetable intake: The moderating role of social support

PubMed Central

McSpadden, Kate E.; Patrick, Heather; Oh, April Y.; Yaroch, Amy L.; Dwyer, Laura A.; Nebeling, Linda C.

2015-01-01

Despite knowing that fruit and vegetable (FV) intake promotes health and well-being, few U.S. adults meet current guidelines. Thus, understanding people’s motivation for FV intake is important for predicting dietary behavior. Applying self-determination theory, the goal of this study was to examine the role of social support as a potential moderator of the link between autonomous and controlled motivations and FV intake. Cross-sectional data from 2,959 adults in the United States were analyzed. Autonomous motivation and perceived social support were positively associated with FV intake, while controlled motivation was negatively associated with FV intake. Additionally, there was evidence that the negative association between controlled motivation and FV intake was attenuated by higher levels of perceived social support. Findings suggest the need for a more comprehensive approach to understanding the role of motivation in health behaviors like FV intake and the potential roles played by friends and family in these motivational processes. PMID:26321416

The association between motivation and fruit and vegetable intake: The moderating role of social support.

PubMed

McSpadden, Kate E; Patrick, Heather; Oh, April Y; Yaroch, Amy L; Dwyer, Laura A; Nebeling, Linda C

2016-01-01

Despite knowing that fruit and vegetable (FV) intake promotes health and well-being, few U.S. adults meet current guidelines. Thus, understanding people's motivation for FV intake is important for predicting dietary behavior. Applying self-determination theory, the goal of this study was to examine the role of social support as a potential moderator of the link between autonomous and controlled motivations and FV intake. Cross-sectional data from 2959 adults in the United States were analyzed. Autonomous motivation and perceived social support were positively associated with FV intake, while controlled motivation was negatively associated with FV intake. Additionally, there was evidence that the negative association between controlled motivation and FV intake was attenuated by higher levels of perceived social support. Findings suggest the need for a more comprehensive approach to understanding the role of motivation in health behaviors like FV intake and the potential roles played by friends and family in these motivational processes. Copyright © 2015 Elsevier Ltd. All rights reserved.

Carboxy-terminal cleavage of the human foamy virus Gag precursor molecule is an essential step in the viral life cycle.

PubMed Central

Enssle, J; Fischer, N; Moebes, A; Mauer, B; Smola, U; Rethwilm, A

1997-01-01

Foamy viruses (FVs) express the Gag protein as a precursor with a molecular mass of 74 kDa (pr74) from which a 70-kDa protein (p70) is cleaved by the viral protease. To gain a better understanding of FV Gag protein processing and function, we have generated and analyzed mutants in the C-terminal gag region of an infectious molecular clone. Our results show that p70 is an N-terminal cleavage product of pr74. However, we were unable to identify a p4 molecule. A virus mutant expressing p70 only was found to be replication competent, albeit at very low titers compared to those of wild-type virus. A strong tendency to synthesize and cleave a pr74 molecule was deduced from the occurrence of revertants upon transfection of this mutant. Substitution of the p6gag domain of human immunodeficiency virus type 1 for the p4 domain of FV resulted in a stable chimeric virus which replicated to titers 10 times lower than those of wild-type virus. FV Gag protein was found to be phosphorylated at serine residues. Mutagenesis of serines conserved in the p4 domain had no influence on viral replication in cell culture. The p70/p74 Gag cleavage was found to be required for viral infectivity, since mutagenesis of the putative cleavage site led to replication-incompetent virus. Interestingly, the cleavage site mutants were defective in the intracellular cDNA synthesis of virion DNA, which indicates that correct FV particle formation and the generation of virion DNA are functionally linked. PMID:9311808

Engineering of a recombinant trivalent single-chain variable fragment antibody directed against rabies virus glycoprotein G with improved neutralizing potency.

PubMed

Turki, Imène; Hammami, Akil; Kharmachi, Habib; Mousli, Mohamed

2014-02-01

Human and equine rabies immunoglobulins are currently available for passive immunization against rabies. However, these are hampered by the limited supply and some drawbacks. Advances in antibody engineering have led to overcome issues of clinical applications and to improve the protective efficacy. In the present study, we report the generation of a trivalent single-chain Fv (scFv50AD1-Fd), that recognizes the rabies virus glycoprotein, genetically fused to the trimerization domain of the bacteriophage T4 fibritin, termed 'foldon' (Fd). scFv50AD1-Fd was expressed as soluble recombinant protein in bacterial periplasmic space and purified through affinity chromatography. The molecular integrity and stability were analyzed by polyacrylamide gradient-gel electrophoresis, size-exclusion chromatography and incubation in human sera. The antigen-binding properties of the trimeric scFv were analyzed by direct and competitive-ELISA. Its apparent affinity constant was estimated at 1.4 ± 0.25 × 10(9)M(-1) and was 75-fold higher than its monovalent scFv (1.9 ± 0.68 × 10(7)M(-1)). The scFv50AD1-Fd neutralized rabies virus in a standard in vitro and in vivo neutralization assay. We showed a high neutralization activity up to 75-fold compared with monovalent format and the WHO standard serum. The gain in avidity resulting from multivalency along with an improved biological activity makes the trivalent scFv50AD1-Fd construct an important reagent for rabies protection. The antibody engineering approach presented here may serve as a strategy for designing a new generation of anti-rabies for passive immunotherapy. Copyright © 2013 Elsevier Ltd. All rights reserved.

An efficient method for variable region assembly in the construction of scFv phage display libraries using independent strand amplification

PubMed Central

Sotelo, Pablo H.; Collazo, Noberto; Zuñiga, Roberto; Gutiérrez-González, Matías; Catalán, Diego; Ribeiro, Carolina Hager; Aguillón, Juan Carlos; Molina, María Carmen

2012-01-01

Phage display library technology is a common method to produce human antibodies. In this technique, the immunoglobulin variable regions are displayed in a bacteriophage in a way that each filamentous virus displays the product of a single antibody gene on its surface. From the collection of different phages, it is possible to isolate the virus that recognizes specific targets. The most common form in which to display antibody variable regions in the phage is the single chain variable fragment format (scFv), which requires assembly of the heavy and light immunoglobulin variable regions in a single gene. In this work, we describe a simple and efficient method for the assembly of immunoglobulin heavy and light chain variable regions in a scFv format. This procedure involves a two-step reaction: (1) DNA amplification to produce the single strand form of the heavy or light chain gene required for the fusion; and (2) mixture of both single strand products followed by an assembly reaction to construct a complete scFv gene. Using this method, we produced 6-fold more scFv encoding DNA than the commonly used splicing by overlap extension PCR (SOE-PCR) approach. The scFv gene produced by this method also proved to be efficient in generating a diverse scFv phage display library. From this scFv library, we obtained phages that bound several non-related antigens, including recombinant proteins and rotavirus particles. PMID:22692130

Fusion Peptide Improves Stability and Bioactivity of Single Chain Antibody against Rabies Virus.

PubMed

Xi, Hualong; Zhang, Kaixin; Yin, Yanchun; Gu, Tiejun; Sun, Qing; Shi, Linqing; Zhang, Renxia; Jiang, Chunlai; Kong, Wei; Wu, Yongge

2017-04-28

The combination of rabies immunoglobulin (RIG) with a vaccine is currently effective against rabies infections, but improvements are needed. Genetic engineering antibody technology is an attractive approach for developing novel antibodies to replace RIG. In our previous study, a single-chain variable fragment, scFv57R, against rabies virus glycoprotein was constructed. However, its inherent weak stability and short half-life compared with the parent RIG may limit its diagnostic and therapeutic application. Therefore, an acidic tail of synuclein (ATS) derived from the C-terminal acidic tail of human alpha-synuclein protein was fused to the C-terminus of scFv57R in order to help it resist adverse stress and improve the stability and halflife. The tail showed no apparent effect on the preparation procedure and affinity of the protein, nor did it change the neutralizing potency in vitro. In the ELISA test of molecular stability, the ATS fusion form of the protein, scFv57R-ATS, showed an increase in thermal stability and longer half-life in serum than scFv57R. The protection against fatal rabies virus challenge improved after fusing the tail to the scFv, which may be attributed to the improved stability. Thus, the ATS fusion approach presented here is easily implemented and can be used as a new strategy to improve the stability and half-life of engineered antibody proteins for practical applications.

Examining neighborhood and interpersonal norms and social support on fruit and vegetable intake in low-income communities.

PubMed

Dulin, Akilah; Risica, Patricia M; Mello, Jennifer; Ahmed, Rashid; Carey, Kate B; Cardel, Michelle; Howe, Chanelle J; Nadimpalli, Sarah; Gans, Kim M

2018-04-05

We examined whether neighborhood-, friend-, and family- norms and social support for consumption and purchase of fruits and vegetables (F&V) were associated with F&V intake among low-income residents in subsidized housing communities. We examined baseline data from a study ancillary to the Live Well/Viva Bien intervention. Participants included 290 residents in four low-income subsidized housing sites who were ≥ 18 years of age, English and/or Spanish speaking, and without medical conditions that prevented consumption of F&V. Linear regression models examined associations of norms and social support with F&V intake after adjustments for sociodemographic characteristics. In the analysis, neighborhood social support for F&V was associated with a 0.31 cup increase in F&V intake (95% CI = 0.05, 0.57). The family norm for eating F&V and family social support for eating F&V were associated with a 0.32 cup (95% CI = 0.13, 0.52) and 0.42 cup (95% CI = 0.19, 0.64) increase in F&V intake, respectively. To our knowledge, no other studies have examined neighborhood, family, and peer norms and social support simultaneously and in relation to F&V intake. These findings may inform neighborhood interventions and community-level policies to reduce neighborhood disparities in F&V consumption.

Generation and characterization of protective antibodies to Marburg virus.

PubMed

Froude, Jeffrey W; Pelat, Thibaut; Miethe, Sebastian; Zak, Samantha E; Wec, Anna Z; Chandran, Kartik; Brannan, Jennifer Mary; Bakken, Russell R; Hust, Michael; Thullier, Philippe; Dye, John M

Marburg virus (MARV) and Ebola virus (EBOV) have been a source of epidemics and outbreaks for several decades. We present here the generation and characterization of the first protective antibodies specific for wild-type MARV. Non-human primates (NHP), cynomolgus macaques, were immunized with viral-replicon particles expressing the glycoproteins (GP) of MARV (Ci67 isolate). An antibody fragment (single-chain variable fragment, scFv) phage display library was built after four immunogen injections, and screened against the GP 1-649 of MARV. Sequencing of 192 selected clones identified 18 clones with distinct V H and V L sequences. Four of these recombinant antibodies (R4A1, R4B11, R4G2, and R3F6) were produced in the scFv-Fc format for in vivo studies. Mice that were challenged with wild-type Marburg virus (Ci67 isolate) receiving 100 µg of scFv-Fc on days -1, 1 and 3 demonstrated protective efficacies ranging from 75-100%. The amino-acid sequences of the scFv-Fcs are similar to those of their human germline counterparts, sharing an identity ranging between 68 and 100% to human germline immunoglobulin. These results demonstrate for the first time that recombinant antibodies offer protection against wild-type MARV, and suggest they may be promising candidates for further therapeutic development especially due to their human homology.

Production of recombinant scFv against p24 of human immunodeficiency virus type 1 by phage display technology.

PubMed

Mohammadzadeh, Sara; Rajabibazl, Masoumeh; Fourozandeh, Mehdi; Rasaee, Mohammad Javad; Rahbarizadeh, Fatemeh; Mohammadi, Mohammad

2014-02-01

Phage display has a fundamental role in protein isolation and engineering. Isolated proteins produced with this method can be modified for specific binding and affinity. P24 is the most produced protein during human immune deficiency virus (HIV) replication; especially in the early steps of HIV-1 infection, its evaluation may have diagnostic values. To test the HIV-1 infection, p24 antigen assay appears to be a very promising alternative to RNA assays. In this study, we have generated a recombinant mouse single chain antibody fragment against p24 of the HIV-1 with the use of phage display technology. After isolation of antibody variable-region (V) gene of B cells extracted from the spleen of an immunized mouse, a library of single chain Fv fragments (scFv) was constructed. The library was used in a series of bio-panning processes against recombinant p24 protein expressed from Escherichia coli. The isolated scFv antibody specifically recognizes the HIV-1 capsid protein p24. The affinity constant of the isolated scFv antibody (MF85) was found to be 2×10(-9) M. Our studies showed that the MF85 scFV antibody has similar properties as that of monoclonal antibodies produced by the hybridoma technology.

Generation of a mouse scFv library specific for porcine aminopeptidase N using the T7 phage display system.

PubMed

Sun, Dongbo; Shi, Hongyan; Chen, Jianfei; Shi, Da; Zhu, Qinghe; Zhang, Hong; Liu, Shengwang; Wang, Yunfeng; Qiu, Huaji; Feng, Li

2012-06-01

Porcine aminopeptidase N (pAPN) is a common cellular receptor for swine transmissible gastroenteritis virus (TGEV) and porcine epidemic diarrhea virus (PEDV). To investigate single-chain fragment variable (scFv) repertoire against pAPN, the genes encoding the immunoglobulin light chain variable region (VL) and heavy chain variable region (VH) were amplified by reverse transcript polymerase chain reaction (RT-PCR) using a series of degenerate primers from the spleen of BABL/c mice immunized with native pAPN. The VL and VH amplicons were combined randomly by a 12 amino acid flexible linker by splicing by overlap extension PCR (SOE-PCR), which produced the scFv gene repertoire. After ligation of the scFv gene repertoire into the T7Select10-3b vector, a mouse scFv phage library specific for pAPN was produced through in vitro packaging. The primary scFv library against pAPN contained 2.0×10(7) recombinant phage clones, and the titer of the amplified library was 3.6×10(9)pfu/mL. BstNI restriction analysis and DNA sequencing revealed that 28 phage clones from the primary pAPN scFv library showed excellent diversity. The effectiveness of the scFv library against pAPN was verified further by phage ELISA using the recombinant protein of the pAPN C subunit as coating antigen. The construction and evaluation of a murine scFv library against the common receptor pAPN of porcine coronaviruses TGEV and PEDV using the T7 phage display system are described. Copyright © 2012 Elsevier B.V. All rights reserved.

Social, attitudinal and behavioural correlates of fruit and vegetable consumption among Cypriot adolescents.

PubMed

Loucaides, Constantinos A; Jago, Russell; Theophanous, Maria

2011-12-01

To examine the prevalence and correlates of fruit and vegetable (FV) consumption in Cypriot adolescents. A cross-sectional study. The Republic of Cyprus. A total of 1966 adolescents with a mean age of 14·7 (SD 2·2) years from nine elementary (n 448), six middle (n 657), five high (n 475) and five technical/vocational schools (n 386) in Cyprus. Participants completed a questionnaire assessing FV consumption using a two-item screening measure and a number of social, attitudinal and behavioural correlates of FV consumption. Overall, 19·3% of adolescents reported consuming five or more portions of FV daily, with elementary and middle school students more likely to meet recommendations (23·8% and 24·4%, respectively) compared with high and technical/vocational school students (14·0% and 12·5%, respectively). Consuming five or more portions of FV was associated with preference for FV (OR = 2·2), family eating patterns (OR = 1·5), friends' FV consumption (OR = 1·2) and school support for FV consumption (OR = 0·8). Consuming at least one portion of fruit daily was significantly associated with preference for FV (OR = 2·0) and family eating patterns (OR = 1·7). Consuming at least one portion of vegetables daily was associated with preference for FV (OR = 4·2) and eating while watching television (OR = 0·8). Targeting individual and family-based components may enhance the effectiveness of intervention programmes to promote FV consumption.

The Molecular Biology of Frog Virus 3 and other Iridoviruses Infecting Cold-Blooded Vertebrates

PubMed Central

Chinchar, V. Gregory; Yu, Kwang H.; Jancovich, James K.

2011-01-01

Frog virus 3 (FV3) is the best characterized member of the family Iridoviridae. FV3 study has provided insights into the replication of other family members, and has served as a model of viral transcription, genome replication, and virus-mediated host-shutoff. Although the broad outlines of FV3 replication have been elucidated, the precise roles of most viral proteins remain unknown. Current studies using knock down (KD) mediated by antisense morpholino oligonucleotides (asMO) and small, interfering RNAs (siRNA), knock out (KO) following replacement of the targeted gene with a selectable marker by homologous recombination, ectopic viral gene expression, and recombinant viral proteins have enabled researchers to systematically ascertain replicative- and virulence-related gene functions. In addition, the application of molecular tools to ecological studies is providing novel ways for field biologists to identify potential pathogens, quantify infections, and trace the evolution of ecologically important viral species. In this review, we summarize current studies using not only FV3, but also other iridoviruses infecting ectotherms. As described below, general principles ascertained using FV3 served as a model for the family, and studies utilizing other ranaviruses and megalocytiviruses have confirmed and extended our understanding of iridovirus replication. Collectively, these and future efforts will elucidate molecular events in viral replication, intrinsic and extrinsic factors that contribute to disease outbreaks, and the role of the host immune system in protection from disease. PMID:22069524

Tetherin promotes the innate and adaptive cell-mediated immune response against retrovirus infection in vivo.

PubMed

Li, Sam X; Barrett, Bradley S; Heilman, Karl J; Messer, Ronald J; Liberatore, Rachel A; Bieniasz, Paul D; Kassiotis, George; Hasenkrug, Kim J; Santiago, Mario L

2014-07-01

Tetherin/BST-2 is a host restriction factor that could directly inhibit retroviral particle release by tethering nascent virions to the plasma membrane. However, the immunological impact of Tetherin during retrovirus infection remains unknown. We now show that Tetherin influences antiretroviral cell-mediated immune responses. In contrast to the direct antiviral effects of Tetherin, which are dependent on cell surface expression, the immunomodulatory effects are linked to the endocytosis of the molecule. Mice encoding endocytosis-competent C57BL/6 Tetherin exhibited lower viremia and pathology at 7 d postinfection with Friend retrovirus (FV) compared with mice encoding endocytosis-defective NZW/LacJ Tetherin. Notably, antiretroviral protection correlated with stronger NK cell responses. In addition, Friend retrovirus infection levels were significantly lower in wild-type C57BL/6 mice than in Tetherin knockout mice at 2 wk postinfection, and antiretroviral protection correlated with stronger NK cell and virus-specific CD8+ T cell responses. The results demonstrate that Tetherin acts as a modulator of the cell-mediated immune response against retrovirus infection in vivo.

Barriers and facilitators to improve fruit and vegetable intake among WIC eligible pregnant Latinas: An application of the Health Action Process Approach framework

PubMed Central

Hromi-Fiedler, Amber; Chapman, Donna; Segura-Pérez, Sofia; Damio, Grace; Clark, Pamela; Martinez, Josefa L.; Pérez-Escamilla, Rafael

2016-01-01

Objective Identify barriers and facilitators to improve prenatal fruit and vegetable (F&V) intake among WIC eligible Latinas using the Health Action Process Approach framework. Design Qualitative data were collected via audiotaped in-depth interviews as part of a larger study to design an intervention to increase prenatal F&V intake. Setting Hartford, Connecticut. Participants Forty-five WIC eligible Latinas completed the study. Included women were: a) ≥ 18 years old; b) in 2nd or 3rd trimester; c) having a singleton pregnancy; d) overweight or obese (i.e. pregravid BMI ≥ 25); e) not on a restricted diet; h) nonsmokers. Phenomenon of Interest Prenatal factors that promote and hinder F&V intake. Analysis Transcripts were independently read, coded, and consensus was reached about emerging themes. Results Ten factors influenced prenatal F&V intake: i) social support, ii) family structure, iii) F&V access, iv) F&V preferences, v) F&V knowledge, vi) F&V health outcome expectations, vii) self-efficacy, viii) intentions, ix) F&V action/coping planning strategies, and x) maternal health status. Conclusions and Implications Social support from family/friends emerged as the primary distal factor driving prenatal F&V intake. Interventions designed to empower pregnant Latinas to gain the access, confidence, knowledge, and strategies necessary to consume more F&Vs need to consider strengthening support to achieve the desired outcome. PMID:27373861

A creative-bonding intervention and a friendly visit approach to promote nursing students' self-transcendence and positive attitudes toward elders: a pilot study.

PubMed

Walsh, Sandra M; Chen, Shiue; Hacker, Marcia; Broschard, Dawn

2008-04-01

Nursing students' disinterest in caring for elders presents health care challenges. As the aged population increases, nursing faculty are challenged to improve students' attitudes toward elder care. Reed's self-transcendence theory guided this pilot study with nursing students (n=22) who implemented either a Creative-Bonding Intervention (CBI) or a Friendly Visit (FV) at senior citizen centers to test the effect of creative approaches on student self-transcendence and attitudes toward elders. Demographic data, a revised Kogan's Attitudes toward Old People statements, and Reed's Self-transcendence Scale were analyzed with descriptive, paired t test, ANCOVA, and Pearson correlation statistics. Results demonstrated significant differences in attitudes in the FV and changes in the expected directions in the CBI group. Self-transcendence had no significant changes. Valuable information was provided by students' comments about the interventions. Reed's belief that self-transcendence is present regardless of age was supported. Future studies are suggested with an increased sample size, a combined CBI/FV intervention, and supportive help during students' intervention delivery.

Activation of the N-Terminally Truncated Form of the Stk Receptor Tyrosine Kinase Sf-Stk by Friend Virus-Encoded gp55 Is Mediated by Cysteine Residues in the Ecotropic Domain of gp55 and the Extracellular Domain of Sf-Stk ▿

PubMed Central

He, Shihan; Ni, Shuang; Hegde, Shailaja; Wang, Xin; Sharda, Daniel R.; August, Avery; Paulson, Robert F.; Hankey, Pamela A.

2010-01-01

Friend virus induces an erythroleukemia in susceptible mice that is initiated by the interaction of the Friend virus-encoded glycoprotein gp55 with the erythropoietin (Epo) receptor and the product of the host Fv2 gene, a naturally occurring truncated form of the Stk receptor tyrosine kinase (Sf-Stk). We have previously demonstrated that the activation of Sf-Stk, recruitment of a Grb2/Gab2/Stat3 signaling complex, and induction of Pu.1 expression by Stat3 are required for the development of the early stage of Friend disease both in vitro and in vivo. Here we demonstrate that the interaction of gp55 with Sf-Stk is dependent on cysteine residues in the ecotropic domain of gp55 and the extracellular domain of Sf-Stk. Point mutation of these cysteine residues or deletion of these domains inhibits the ability of gp55 to interact with Sf-Stk, resulting in the inability of these proteins to promote the Epo-independent growth of erythroid progenitor cells. We also demonstrate that the interaction of gp55 with Sf-Stk does not promote dimerization of Sf-Stk but results in enhanced phosphorylation of Sf-Stk and the relocalization of Sf-Stk from the cytosol to the plasma membrane. Finally, we demonstrate that a constitutively active form of Sf-Stk (Sf-StkM330T), as well as its human counterpart, Sf-Ron, promotes Epo-independent colony formation in the absence of gp55 and that this response is also dependent on the cysteines in the extracellular domains of Sf-StkM330T and Sf-Ron. These data suggest that the cysteines in the extracellular domains of Sf-Stk and Sf-Ron may also mediate the interaction of these truncated receptors with other cellular factors that regulate their ability to promote cytokine-independent growth. PMID:20016000

Molecular characterization of iridoviruses isolated from sympatric amphibians and fish

USGS Publications Warehouse

Mao, J.; Green, D.E.; Fellers, G.; Chinchar, V.G.

1999-01-01

Iridoviruses infect invertebrates (primarily insects and crustaceans) and ectothermic vertebrates (fish, amphibians, and reptiles). Identical, or nearly identical viruses, have been isolated from different animals within the same taxonomic class, indicating that infection by a given virus is not limited to a single species. Although inter-class infections have been documented following experimental infection with vertebrate iridoviruses, it is not clear whether such infections occur in nature. Here we report the isolation of apparently identical iridoviruses from wild sympatric fish (the threespine stickleback, Gasterostelus aculeatus) and amphibians (the red-legged frog, Rana aurora). Viruses isolated from sticklebacks (stickleback virus, SBV) and from a red-legged frog tadpole (tadpole virus 2, TV2) replicated in fathead minnow (FHM) cells and synthesized proteins which co-migrated with those of frog virus 3 (FV3). Following restriction endonuclease digestion of viral DNA with Hind III and Xba I, gel analysis showed that the profiles of SBV and TV2 were identical to each other and distinct from FV3. Using oligonucleotide primers specific for a highly conserved region of the iridovirus major capsid protein, an not, vert, ~500 nucleotide DNA fragment was amplified from SBV and TV2. Sequence analysis showed that within this 500 nucleotide region SBV and TV2 were identical to each other and to FV3. Taken together these results provide the first evidence that iridoviruses naturally infect animals belonging to different taxonomic classes, and strengthen the suggestion that fish may serve as a reservoir for amphibian viruses or vice versa.

Glycosylphosphatidylinositol-Anchored Anti-HIV scFv Efficiently Protects CD4 T Cells from HIV-1 Infection and Deletion in hu-PBL Mice

PubMed Central

Ye, Chaobaihui; Wang, Weiming; Cheng, Liang; Li, Guangming; Wen, Michael; Wang, Qi; Zhang, Qing; Li, Dan

2016-01-01

ABSTRACT Despite success in viral inhibition and CD4 T cell recovery by highly active antiretroviral treatment (HAART), HIV-1 is still not curable due to the persistence of the HIV-1 reservoir during treatment. One patient with acute myeloid leukemia who received allogeneic hematopoietic stem cell transplantation from a homozygous CCR5 Δ32 donor has had no detectable viremia for 9 years after HAART cessation. This case has inspired a field of HIV-1 cure research focusing on engineering HIV-1 resistance in permissive cells. Here, we employed a glycosylphosphatidylinositol (GPI)-scFv X5 approach to confer resistance of human primary CD4 T cells to HIV-1. We showed that primary CD4 T cells expressing GPI-scFv X5 were resistant to CCR5 (R5)-, CXCR4 (X4)-, and dual-tropic HIV-1 and had a survival advantage compared to control cells ex vivo. In a hu-PBL mouse study, GPI-scFv X5-transduced CD4 T cells were selected in peripheral blood and lymphoid tissues upon HIV-1 infection. Finally, GPI-scFv X5-transduced CD4 T cells, after being cotransfused with HIV-infected cells, showed significantly reduced viral loads and viral RNA copy numbers relative to CD4 cells in hu-PBL mice compared to mice with GPI-scFv AB65-transduced CD4 T cells. We conclude that GPI-scFv X5-modified CD4 T cells could potentially be used as a genetic intervention against both R5- and X4-tropic HIV-1 infections. IMPORTANCE Blocking of HIV-1 entry is one of most promising approaches for therapy. Genetic disruption of the HIV-1 coreceptor CCR5 by nucleases in T cells is under 2 clinical trials and leads to reduced viremia in patients. However, the emergence of viruses using the CXCR4 coreceptor is a concern for therapies applying single-coreceptor disruption. Here, we report that HIV-1-permissive CD4 T cells engineered with GPI-scFv X5 are resistant to R5-, X4-, or dual-tropic virus infection ex vivo. In a preclinical study using hu-PBL mice, we show that CD4 T cells were protected and that GPI-scFv X5-transduced cells were selected in HIV-1-infected animals. Moreover, we show that GPI-scFv X5-transduced CD4 T cells exerted a negative effect on virus replication in vivo. We conclude that GPI-scFv X5-modified CD4 T cells could potentially be used as a genetic intervention against both R5- and X4-tropic HIV-1 infections. PMID:27881659

Amphibian ocular malformation associated with frog virus 3.

PubMed

Burton, Elizabeth C; Miller, Debra L; Styer, Eloise L; Gray, Matthew J

2008-09-01

During an on-going amphibian ecology study, a free-ranging American bullfrog (Rana catesbeiana) metamorph was captured in a pitfall trap adjacent to a constructed farm pond at the Plateau Research and Education Center (PREC) on the Cumberland Plateau near Crossville, Tennessee, USA. Grossly, the right eye was approximately 50% the size of the left. Stereo and light microscopic examination revealed two granulomas within the orbit. Electron microscopic examination revealed virus particles scattered throughout one structure but mostly aggregated toward the center. Subsequent PCR and sequencing (GenBank accession Number EF175670) confirmed frog virus 3 (FV3). This represents the first report of a malformation in an anuran associated with FV3.

Vectorization in an oncolytic vaccinia virus of an antibody, a Fab and a scFv against programmed cell death -1 (PD-1) allows their intratumoral delivery and an improved tumor-growth inhibition.

PubMed

Kleinpeter, Patricia; Fend, Laetitia; Thioudellet, Christine; Geist, Michel; Sfrontato, Nathalie; Koerper, Véronique; Fahrner, Catherine; Schmitt, Doris; Gantzer, Murielle; Remy-Ziller, Christelle; Brandely, Renée; Villeval, Dominique; Rittner, Karola; Silvestre, Nathalie; Erbs, Philippe; Zitvogel, Laurence; Quéméneur, Eric; Préville, Xavier; Marchand, Jean-Baptiste

2016-01-01

We report here the successful vectorization of a hamster monoclonal IgG (namely J43) recognizing the murine Programmed cell death-1 (mPD-1) in Western Reserve (WR) oncolytic vaccinia virus. Three forms of mPD-1 binders have been inserted into the virus: whole antibody (mAb), Fragment antigen-binding (Fab) or single-chain variable fragment (scFv). MAb, Fab and scFv were produced and assembled with the expected patterns in supernatants of cells infected by the recombinant viruses. The three purified mPD-1 binders were able to block the binding of mPD-1 ligand to mPD-1 in vitro . Moreover, mAb was detected in tumor and in serum of C57BL/6 mice when the recombinant WR-mAb was injected intratumorally (IT) in B16F10 and MCA 205 tumors. The concentration of circulating mAb detected after IT injection was up to 1,900-fold higher than the level obtained after a subcutaneous (SC) injection (i.e., without tumor) confirming the virus tropism for tumoral cells and/or microenvironment. Moreover, the overall tumoral accumulation of the mAb was higher and lasted longer after IT injection of WR-mAb1, than after IT administration of 10 µg of J43. The IT injection of viruses induced a massive infiltration of immune cells including activated lymphocytes (CD8 + and CD4 + ). Interestingly, in the MCA 205 tumor model, WR-mAb1 and WR-scFv induced a therapeutic control of tumor growth similar to unarmed WR combined to systemically administered J43 and superior to that obtained with an unarmed WR. These results pave the way for next generation of oncolytic vaccinia armed with immunomodulatory therapeutic proteins such as mAbs.

Vectorization in an oncolytic vaccinia virus of an antibody, a Fab and a scFv against programmed cell death -1 (PD-1) allows their intratumoral delivery and an improved tumor-growth inhibition

PubMed Central

Kleinpeter, Patricia; Fend, Laetitia; Thioudellet, Christine; Geist, Michel; Sfrontato, Nathalie; Koerper, Véronique; Fahrner, Catherine; Schmitt, Doris; Gantzer, Murielle; Remy-Ziller, Christelle; Brandely, Renée; Villeval, Dominique; Rittner, Karola; Silvestre, Nathalie; Erbs, Philippe; Zitvogel, Laurence; Quéméneur, Eric; Préville, Xavier; Marchand, Jean-Baptiste

2016-01-01

ABSTRACT We report here the successful vectorization of a hamster monoclonal IgG (namely J43) recognizing the murine Programmed cell death-1 (mPD-1) in Western Reserve (WR) oncolytic vaccinia virus. Three forms of mPD-1 binders have been inserted into the virus: whole antibody (mAb), Fragment antigen-binding (Fab) or single-chain variable fragment (scFv). MAb, Fab and scFv were produced and assembled with the expected patterns in supernatants of cells infected by the recombinant viruses. The three purified mPD-1 binders were able to block the binding of mPD-1 ligand to mPD-1 in vitro. Moreover, mAb was detected in tumor and in serum of C57BL/6 mice when the recombinant WR-mAb was injected intratumorally (IT) in B16F10 and MCA 205 tumors. The concentration of circulating mAb detected after IT injection was up to 1,900-fold higher than the level obtained after a subcutaneous (SC) injection (i.e., without tumor) confirming the virus tropism for tumoral cells and/or microenvironment. Moreover, the overall tumoral accumulation of the mAb was higher and lasted longer after IT injection of WR-mAb1, than after IT administration of 10 µg of J43. The IT injection of viruses induced a massive infiltration of immune cells including activated lymphocytes (CD8+ and CD4+). Interestingly, in the MCA 205 tumor model, WR-mAb1 and WR-scFv induced a therapeutic control of tumor growth similar to unarmed WR combined to systemically administered J43 and superior to that obtained with an unarmed WR. These results pave the way for next generation of oncolytic vaccinia armed with immunomodulatory therapeutic proteins such as mAbs. PMID:27853644

Mechanism of HSV infection through soluble adapter-mediated virus bridging to the EGF receptor

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

Nakano, Kenji, E-mail: kenakano@med.kyushu-u.ac.j; Kobayashi, Masatoshi; Nakamura, Kei-ichiro

2011-04-25

Herpes simplex virus entry into cells requires the binding of envelope glycoprotein D (gD) to an entry receptor. Depending on the cell, entry occurs by different mechanisms, including fusion at the cell surface or endocytosis. Here we examined the entry mechanism through a non-HSV receptor mediated by a soluble bi-specific adapter protein composed of recognition elements for gD and the EGF receptor (EGFR). Virus entered into endosomes using either EGF or an EGFR-specific single chain antibody (scFv) for receptor recognition. Infection was less efficient with the EGF adapter which could be attributed to its weaker binding to a viral gD.more » Infection mediated by the scFv adapter was pH sensitive, indicating that gD-EGFR bridging alone was insufficient for capsid release from endosomes. We also show that the scFv adapter enhanced infection of EGFR-expressing tumor tissue in vivo. Our results indicate that adapters may retarget HSV infection without drastically changing the entry mechanism.« less

Structural and functional characterization of an anti-West Nile virus monoclonal antibody and its single-chain variant produced in glycoengineered plants

PubMed Central

Lai, Huafang; He, Junyun; Hurtado, Jonathan; Stahnke, Jake; Fuchs, Anja; Mehlhop, Erin; Gorlatov, Sergey; Loos, Andreas; Diamond, Michael S.; Chen, Qiang

2014-01-01

Previously, our group engineered a plant-derived monoclonal antibody (MAb pE16) that efficiently treated West Nile virus (WNV) infection in mice. In this study, we developed a pE16 variant consisting of a single-chain variable fragment (scFv) fused to the heavy chain constant domains (CH) of human IgG (pE16scFv-CH). pE16 and pE16scFv-CH were expressed and assembled efficiently in Nicotiana benthamiana ΔXF plants, a glycosylation mutant lacking plant specific N-glycan residues. Glycan analysis revealed that ΔXF plant-derived pE16scFv-CH (ΔXFpE16scFv-CH) and pE16 (ΔXFpE16) both displayed a mammalian glycosylation profile. ΔXFpE16 and ΔXFpE16scFv-CH demonstrated equivalent antigen binding affinity and kinetics, and slightly enhanced neutralization of WNV in vitro compared to the parent mammalian cell-produced E16 (mE16). A single dose of ΔXFpE16 or ΔXFpE16scFv-CH protected mice against WNV-induced mortality even 4 days after infection at equivalent rates as mE16. This study provides a detailed tandem comparison of the expression, structure and function of a therapeutic MAb and its single-chain variant produced in glycoengineered plants. Moreover, it demonstrates the development of anti-WNV MAb therapeutic variants that are equivalent in efficacy to pE16, simpler to produce, and likely safer to use as therapeutics due to their mammalian N-glycosylation. This platform may lead to a more robust and cost effective production of antibody-based therapeutics against WNV infection and other infectious, inflammatory, or neoplastic diseases. PMID:24975464

Evolution of Foamy Viruses: The Most Ancient of All Retroviruses †

PubMed Central

Rethwilm, Axel; Bodem, Jochen

2013-01-01

Recent evidence indicates that foamy viruses (FVs) are the oldest retroviruses (RVs) that we know and coevolved with their hosts for several hundred million years. This coevolution may have contributed to the non-pathogenicity of FVs, an important factor in development of foamy viral vectors in gene therapy. However, various questions on the molecular evolution of FVs remain still unanswered. The analysis of the spectrum of animal species infected by exogenous FVs or harboring endogenous FV elements in their genome is pivotal. Furthermore, animal studies might reveal important issues, such as the identification of the FV in vivo target cells, which than require a detailed characterization, to resolve the molecular basis of the accuracy with which FVs copy their genome. The issues of the extent of FV viremia and of the nature of the virion genome (RNA vs. DNA) also need to be experimentally addressed. PMID:24072062

Xenopus-FV3 host-pathogen interactions and immune evasion.

PubMed

Jacques, Robert; Edholm, Eva-Stina; Jazz, Sanchez; Odalys, Torres-Luquis; Francisco, De Jesús Andino

2017-11-01

We first review fundamental insights into anti-ranavirus immunity learned with the Xenopus laevis/ranavirus FV3 model that are generally applicable to ectothermic vertebrates. We then further investigate FV3 genes involved in immune evasion. Focusing on FV3 knockout (KO) mutants defective for a putative viral caspase activation and recruitment domain-containing (CARD)-like protein (Δ64R-FV3), a β-hydroxysteroid dehydrogenase homolog (Δ52L-FV3), and an immediate-early18kDa protein (FV3-Δ18K), we assessed the involvement of these viral genes in replication, dissemination and interaction with peritoneal macrophages in tadpole and adult frogs. Our results substantiate the role of 64R and 52L as critical immune evasion genes, promoting persistence and dissemination in the host by counteracting type III IFN in tadpoles and type I IFN in adult frogs. Comparably, the substantial accumulation of genome copy numbers and exacerbation of type I and III IFN gene expression responses but deficient release of infectious virus suggests that 18K is a viral regulatory gene. Copyright © 2017 Elsevier Inc. All rights reserved.

Interaction of S17 Antibody with the Functional Binding Region of the Hepatitis B Virus Pre-S2 Epitope.

PubMed

Chang, Chang-Yu; Chang, Fu-Ling; Chiang, Chen-Wei; Lo, Yan-Ni; Lin, Tsai-Yu; Chen, Wang-Chuan; Tsai, Keng-Chang; Lee, Yu-Ching

2018-05-30

To understand the mechanism for inhibition of hepatitis B virus (HBV) infection is important. In this study, single-chain variable fragment (scFv) antibodies were generated and directed to the pre-S2 epitope of HBV surface antigen (HBsAg). These human scFvs were isolated from a person with history of HBV infection by phage display technology. An evaluation of panning efficiency revealed that the eluted phage titer was increased, indicating that specific clones were enriched after panning. Selected scFvs were characterized with the recombinant HBsAg through Western blotting and enzyme-linked immunosorbent assay to confirm the binding ability. Flow cytometry analysis and immunocytochemical staining revealed that one scFv, S17, could recognize endogenous HBsAg expressed on the HepG2215 cell membrane. Moreover, the binding affinity of scFv S17 to the pre-S2 epitope was determined to be 4.2 × 10 -8 M. Two ion interactions were observed as the major driving forces for scFv S17 interacting with pre-S2 by performing a rational molecular docking analysis. This study provides insights into the structural basis to understand the interactions between an antibody and the pre-S2 epitope. The functional scFv format can potentially be used in future immunotherapeutic applications.

Foamy Virus Vector-mediated Gene Correction of a Mouse Model of Wiskott–Aldrich Syndrome

PubMed Central

Uchiyama, Toru; Adriani, Marsilio; Jagadeesh, G Jayashree; Paine, Adam; Candotti, Fabio

2012-01-01

The Wiskott–Aldrich syndrome (WAS) is an X-linked disorder characterized by eczema, thrombocytopenia and immunodeficiency. Hematopoietic cell transplantation can cure the disease and gene therapy is being tested as an alternative treatment option. In this study, we assessed the use of foamy virus (FV) vectors as a gene transfer system for WAS, using a Was knockout (KO) mouse model. Preliminary experiments using FV vectors expressing the green fluorescent protein under the transcriptional control of the endogenous WAS promoter or a ubiquitously acting chromatin opening element allowed us to define transduction conditions resulting in high (>40%) and long-term in-vivo marking of blood cells after transplantation. In following experiments, Was KO mice were treated with FV vectors containing the human WAS complementary DNA (cDNA). Transplanted animals expressed the WAS protein (WASp) in T and B lymphocytes, as well as platelets and showed restoration of both T-cell receptor-mediated responses and B-cell migration. We also observed recovery of platelet adhesion and podosome formation in dendritic cells (DCs) of treated mice. These data demonstrate that FV vectors can be effective for hematopoietic stem cell (HSC)-directed gene correction of WAS. PMID:22215016

Isolation and Characterization of an Equine Foamy Virus

PubMed Central

Tobaly-Tapiero, Joelle; Bittoun, Patricia; Neves, Manuel; Guillemin, Marie-Claude; Lecellier, Charles-Henri; Puvion-Dutilleul, Francine; Gicquel, Bernard; Zientara, Stephan; Giron, Marie-Louise; de Thé, Hugues; Saïb, Ali

2000-01-01

Foamy viruses (FVs) are complex retroviruses which have been isolated from different animal species including nonhuman primates, cattle, and cats. Here, we report the isolation and characterization of a new FV isolated from blood samples of horses. Similar to other FVs, the equine foamy virus (EFV) exhibits a highly characteristic ultrastructure and induces syncytium formation and subsequent cell lysis on a large number of cell lines. Molecular cloning of EFV reveals that the general organization is that of other known FVs, whereas sequence similarity with its bovine FV counterpart is only 40%. Interestingly, EFV buds exclusively from the plasma membrane and not from the endoplasmic reticulum (ER), as previously shown for other FVs. The absence of the ER retrieval dilysine motif in EFV Env is likely responsible for this unexpected sorting pathway. PMID:10756018

Novel double-stranded RNA viruses of plant-feeding insects encode a serine-alanine-proline rich protein and a polymerase distantly related to fungal viruses

USDA-ARS?s Scientific Manuscript database

Novel double stranded RNAs (~8 kbp) were isolated from the three cornered alfalfa hopper (Spissistilus festinus) and beet leafhopper (Circulifer tenellus), two plant-feeding hemipteran insect pests. Genome organization of the two new viruses, designated as Spissistilus festinus virus 1 (SpFV1) and ...

Plant-feeding insects harbor double-stranded RNA viruses encoding a novel proline-alanine rich protein and a polymerase distantly related to that of fungal viruses

USDA-ARS?s Scientific Manuscript database

Novel double-stranded RNAs (~8 kbp) were isolated from three cornered alfalfa hopper (Spissistilus festinus) and beet leafhopper (Circulifer tenellus), two plant-feeding hemipteran insect pests. Genomes of the two new viruses, designated as Spissistilus festinus virus 1 (SpFV1) and Circulifer tenell...

Production and characterization of single-chain antibody (scFv) against 3ABC non-structural protein in Escherichia coli for sero-diagnosis of Foot and Mouth Disease virus.

PubMed

Sharma, Gaurav K; Mahajan, Sonalika; Matura, Rakesh; Subramaniam, Saravanan; Mohapatra, Jajati K; Pattnaik, Bramhadev

2014-11-01

Differentiation of Foot-and-Mouth Disease infected from vaccinated animals is essential for effective implementation of vaccination based control programme. Detection of antibodies against 3ABC non-structural protein of FMD virus by immunodiagnostic assays provides reliable indication of FMD infection. Sero-monitoring of FMD in the large country like India is a big task where thousands of serum samples are annually screened. Currently, monoclonal or polyclonal antibodies are widely used in these immunodiagnostic assays. Considering the large population of livestock in the country, an economical and replenishable alternative of these antibodies was required. In this study, specific short chain variable fragment (scFv) antibody against 3B region of 3ABC poly-protein was developed. High level of scFv expression in Escherichia coli system was obtained by careful optimization in four different strains. Two formats of enzyme immunoassays (sandwich and competitive ELISAs) were optimized using scFv with objective to differentiate FMD infected among the vaccinated population. The assays were statistically validated by testing 2150 serum samples. Diagnostic sensitivity/specificity of sandwich and competitive ELISAs were determined by ROC method as 92.2%/95.5% and 89.5%/93.5%, respectively. This study demonstrated that scFv is a suitable alternate for immunodiagnosis of FMD on large scale. Copyright © 2014 The International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.

Recombinant scFv antibodies against infectious pancreatic necrosis virus isolated by flow cytometry.

PubMed

Xu, Li-Ming; Zhao, Jing-Zhuang; Liu, Miao; Cao, Yong-Sheng; Yin, Jia-Sheng; Liu, Hong-Bai; Lu, Tongyan

2016-11-01

Infectious pancreatic necrosis is a significant disease of farmed salmonids in China. In this study, a single chain variable fragment (scFv) antibody library derived from rainbow trout (Oncorhynchus mykiss) and viral protein VP2 of a Chinese infectious pancreatic necrosis virus (IPNV) isolate ChRtm213 were co-expressed by a bacterial display technology. The library was subjected to three rounds of screening by flow cytometry (FCM) to select IPNV specific antibodies. Six antibody clones with different mean fluorescence intensities (MFI) were obtained by picking colonies at random. The antibody clones were expressed and purified. The purified IPNV-specific scFv antibodies were used successfully in Western blotting, enzyme linked immunosorbent assay (ELISA) and an immunofluorescence antibody test (IFAT). This method provides a high throughput means to screen an antibody library by flow cytometry, and isolate a panel of antibody that can be used as potential reagents for the detection and study of IPNV that are prevalent in China. Copyright © 2016 Elsevier B.V. All rights reserved.

Biophysical Properties of Frog Virus and Its Deoxyribonucleic Acid: Fate of Radioactive Virus in the Early Stage of Infection 1

PubMed Central

Smith, William R.; McAuslan, Brian R.

1969-01-01

Frog virus (FV-3) was banded by isopycnic centrifugation in cesium chloride, sucrose, or potassium tartrate. Two bands of infectivity were regularly found at positions in cesium chloride corresponding to densities of 1.26 and 1.30 g/cm3, respectively. Deoxyribonucleic acid from either band had the following characteristics: double-stranded; a Tm of 76.3 C in 0.1 SSC (0.015 m NaCl plus 0.015 m sodium citrate) and a buoyant density of 1.720 g/cm3 in cesium chloride, corresponding to a guanine plus cytosine content of 56 to 58% and a molecular weight of 130 × 106 daltons, determined by velocity sedimentation. These data, together with electron micrographs of sections of cells infected with material from either band suggest that two types of infectious frog virus particles exists, rather than a second virus in the frog virus stocks. The composition of frog virus was determined. It was found that highly purified preparations of frog virus were composed of 55.8% protein, 30.1% deoxyribonucleic acid, and 14.2% lipid. The kinetics of adsorption and uncoating of FV-3 was studied with radioactive virus. Uncoating is comparatively rapid and in contrast to poxvirus is unaffected by inhibitors of protein synthesis. Images PMID:4980848

Pathogenicity of frog virus 3-like virus in red-eared slider turtles (Trachemys scripta elegans) at two environmental temperatures.

PubMed

Allender, M C; Mitchell, M A; Torres, T; Sekowska, J; Driskell, E A

2013-01-01

Ranaviral disease has affected several species of reptiles, but disease progression and mortality in relation to environmental temperature has yet to be determined. In this study, two separate trials challenged adult female red-eared slider turtles (Trachemys scripta elegans) with a ranavirus (frog virus 3-like virus; FV3) isolate at environmental temperatures of 22 °C (n = 4) and 28 °C (n = 4). The mortality rates in the turtles in the 22 °C and 28 °C trials were 100% and 50%, respectively. Median survival time for turtles exposed to FV3 at 22 °C was 24 days, while it was 30 days in the group kept at 28 °C. Consistent microscopical lesions were observed only in the group inoculated at 22 °C and included fibrinoid necrosis of vessels in the spleen, vascular and sinusoidal thrombi in the liver, necrotizing myositis and a mild heterophilic interstitial pneumonia. Quantitative polymerase chain reaction, targeting a conserved portion of the major capsid protein, was able to detect virus copies in whole blood, oral and cloacal swabs, tongue, skeletal muscle, lung, heart, liver, spleen, ovary and kidney. Viral copy number in ante-mortem clinical samples was non-significantly highest in whole blood, while kidney had the highest viral copy number in post-mortem samples. All samples had higher virus copy number in turtles exposed to FV3 at 22 °C compared with 28 °C. This study determined that environmental temperature affects the survival and disease progression in ranavirus-infected red-eared slider turtles, which will aid in managing animals in a clinical or free-ranging setting. Copyright © 2013 Elsevier Ltd. All rights reserved.

Single-Chain Fv-Based Anti-HIV Proteins: Potential and Limitations

PubMed Central

West, Anthony P.; Galimidi, Rachel P.; Gnanapragasam, Priyanthi N. P.

2012-01-01

The existence of very potent, broadly neutralizing antibodies against human immunodeficiency virus type 1 (HIV-1) offers the potential for prophylaxis against HIV-1 infection by passive immunization or gene therapy. Both routes permit the delivery of modified forms of IgGs. Smaller reagents are favored when considering ease of tissue penetration and the limited capacities of gene therapy vectors. Immunoadhesin (single-chain fragment variable [scFv]-Fc) forms of IgGs are one class of relatively small reagent that has been explored for delivery by adeno-associated virus. Here we investigated the neutralization potencies of immunoadhesins compared to those of their parent IgGs. For the antibodies VRC01, PG9, and PG16, the immunoadhesins showed modestly reduced potencies, likely reflecting reduced affinities compared to those of the parent IgG, and the VRC01 immunoadhesin formed dimers and multimers with reduced neutralization potencies. Although scFv forms of neutralizing antibodies may exhibit affinity reductions, they provide a means of building reagents with multiple activities. Attachment of the VRC01 scFv to PG16 IgG yielded a bispecific reagent whose neutralization activity combined activities from both parent antibodies. Although the neutralization activity due to each component was partially reduced, the combined reagent is attractive since fewer strains escaped neutralization. PMID:22013046

Novel insect-specific flavivirus isolated from northern Europe

PubMed Central

Huhtamo, Eili; Moureau, Gregory; Cook, Shelley; Julkunen, Ora; Putkuri, Niina; Kurkela, Satu; Uzcátegui, Nathalie Y.; Harbach, Ralph E.; Gould, Ernest A.; Vapalahti, Olli; de Lamballerie, Xavier

2012-01-01

Mosquitoes collected in Finland were screened for flaviviral RNA leading to the discovery and isolation of a novel flavivirus designated Hanko virus (HANKV). Virus characterization, including phylogenetic analysis of the complete coding sequence, confirmed HANKV as a member of the “insect-specific” flavivirus (ISF) group. HANKV is the first member of this group isolated from northern Europe, and therefore the first northern European ISF for which the complete coding sequence has been determined. HANKV was not transcribed as DNA in mosquito cell culture, which appears atypical for an ISF. HANKV shared highest sequence homology with the partial NS5 sequence available for the recently discovered Spanish Ochlerotatus flavivirus (SOcFV). Retrospective analysis of mitochondrial sequences from the virus-positive mosquito pool suggested an Ochlerotatus mosquito species as the most likely host for HANKV. HANKV and SOcFV may therefore represent a novel group of Ochlerotatus-hosted insect-specific flaviviruses in Europe and further afield. PMID:22999256

Health risks and travel preparation among foreign visitors and expatriates during the 2008 Beijing Olympic and Paralympic Games.

PubMed

Jentes, Emily S; Davis, Xiaohong M; Macdonald, Susan; Snyman, P Johann; Nelson, Hugh; Quarry, Doug; Lai, Irene; van Vliet, Erik W N; Balaban, Victor; Marano, Cinzia; Mues, Katherine; Kozarsky, Phyllis; Marano, Nina

2010-03-01

During the 2008 Olympic and Paralympic Games, we conducted surveillance of illnesses among travelers at six Beijing clinics. Surveys asked about demographic, pre-travel, and vaccination information, and physician-provided diagnoses. Of 807 respondents, 38% and 57% were classified as foreign visitors (FV) and expatriates, respectively. Less than one-half of FV sought pre-travel advice; sources included health-care providers and friends/family. FV vaccination rate was also low; however, most vaccines given were recommended by the Centers for Disease Control and Prevention. The most common FV diagnoses were respiratory, injury/musculoskeletal, and gastrointestinal illnesses; for expatriates, injury/musculoskeletal, respiratory, and dermatologic were the most common illnesses. Respiratory illnesses in expatriates were significantly less in 2008 than during 2004-2007 (chi(2) = 10.2; P = 0.0014), suggesting that control programs may have reduced pollutants/respiratory irritants during the 2008 Games. We found no previous studies of health outcomes among expatriates living in cities with mass travel events. These findings highlight the need to continuously disseminate information to health-care providers advising travelers.

Health Risks and Travel Preparation Among Foreign Visitors and Expatriates During the 2008 Beijing Olympic and Paralympic Games

PubMed Central

Jentes, Emily S.; Davis, Xiaohong M.; MacDonald, Susan; Snyman, P. Johann; Nelson, Hugh; Quarry, Doug; Lai, Irene; van Vliet, Erik W. N.; Balaban, Victor; Marano, Cinzia; Mues, Katherine; Kozarsky, Phyllis; Marano, Nina

2010-01-01

During the 2008 Olympic and Paralympic Games, we conducted surveillance of illnesses among travelers at six Beijing clinics. Surveys asked about demographic, pre-travel, and vaccination information, and physician-provided diagnoses. Of 807 respondents, 38% and 57% were classified as foreign visitors (FV) and expatriates, respectively. Less than one-half of FV sought pre-travel advice; sources included health-care providers and friends/family. FV vaccination rate was also low; however, most vaccines given were recommended by the Centers for Disease Control and Prevention. The most common FV diagnoses were respiratory, injury/musculoskeletal, and gastrointestinal illnesses; for expatriates, injury/musculoskeletal, respiratory, and dermatologic were the most common illnesses. Respiratory illnesses in expatriates were significantly less in 2008 than during 2004–2007 (χ2 = 10.2; P = 0.0014), suggesting that control programs may have reduced pollutants/respiratory irritants during the 2008 Games. We found no previous studies of health outcomes among expatriates living in cities with mass travel events. These findings highlight the need to continuously disseminate information to health-care providers advising travelers. PMID:20207875

Reassessment of murine APOBEC1 as a retrovirus restriction factor in vivo.

PubMed

Barrett, Bradley S; Guo, Kejun; Harper, Michael S; Li, Sam X; Heilman, Karl J; Davidson, Nicholas O; Santiago, Mario L

2014-11-01

APOBEC1 is a cytidine deaminase involved in cholesterol metabolism that has been linked to retrovirus restriction, analogous to the evolutionarily-related APOBEC3 proteins. In particular, murine APOBEC1 was shown to inhibit Friend retrovirus (FV) in vitro, generating high levels of C-to-T and G-to-A mutations. These observations raised the possibility that FV infection might be altered in APOBEC1-null mice. To examine this question directly, we infected wild-type and APOBEC1-null mice with FV complex and evaluated acute infection levels. Surprisingly, APOBEC1-null mice exhibited similar cellular infection levels and plasma viremia relative to wild-type mice. Moreover, next-generation sequencing analyses revealed that in contrast to APOBEC3, APOBEC1 did not enhance retroviral C-to-T and G-to-A mutational frequencies in genomic DNA. Thus, APOBEC1 neither inhibited nor significantly drove the molecular evolution of FV in vivo. Our findings reinforce that not all retrovirus restriction factors characterized as potent in vitro may be functionally relevant in vivo. Copyright © 2014 Elsevier Inc. All rights reserved.

Tetherin/BST-2 promotes dendritic cell activation and function during acute retrovirus infection.

PubMed

Li, Sam X; Barrett, Bradley S; Guo, Kejun; Kassiotis, George; Hasenkrug, Kim J; Dittmer, Ulf; Gibbert, Kathrin; Santiago, Mario L

2016-02-05

Tetherin/BST-2 is a host restriction factor that inhibits retrovirus release from infected cells in vitro by tethering nascent virions to the plasma membrane. However, contradictory data exists on whether Tetherin inhibits acute retrovirus infection in vivo. Previously, we reported that Tetherin-mediated inhibition of Friend retrovirus (FV) replication at 2 weeks post-infection correlated with stronger natural killer, CD4+ T and CD8+ T cell responses. Here, we further investigated the role of Tetherin in counteracting retrovirus replication in vivo. FV infection levels were similar between wild-type (WT) and Tetherin KO mice at 3 to 7 days post-infection despite removal of a potent restriction factor, Apobec3/Rfv3. However, during this phase of acute infection, Tetherin enhanced myeloid dendritic cell (DC) function. DCs from infected, but not uninfected, WT mice expressed significantly higher MHC class II and the co-stimulatory molecule CD80 compared to Tetherin KO DCs. Tetherin-associated DC activation during acute FV infection correlated with stronger NK cell responses. Furthermore, Tetherin+ DCs from FV-infected mice more strongly stimulated FV-specific CD4+ T cells ex vivo compared to Tetherin KO DCs. The results link the antiretroviral and immunomodulatory activity of Tetherin in vivo to improved DC activation and MHC class II antigen presentation.

Tetherin/BST-2 promotes dendritic cell activation and function during acute retrovirus infection

PubMed Central

Li, Sam X.; Barrett, Bradley S.; Guo, Kejun; Kassiotis, George; Hasenkrug, Kim J.; Dittmer, Ulf; Gibbert, Kathrin; Santiago, Mario L.

2016-01-01

Tetherin/BST-2 is a host restriction factor that inhibits retrovirus release from infected cells in vitro by tethering nascent virions to the plasma membrane. However, contradictory data exists on whether Tetherin inhibits acute retrovirus infection in vivo. Previously, we reported that Tetherin-mediated inhibition of Friend retrovirus (FV) replication at 2 weeks post-infection correlated with stronger natural killer, CD4+ T and CD8+ T cell responses. Here, we further investigated the role of Tetherin in counteracting retrovirus replication in vivo. FV infection levels were similar between wild-type (WT) and Tetherin KO mice at 3 to 7 days post-infection despite removal of a potent restriction factor, Apobec3/Rfv3. However, during this phase of acute infection, Tetherin enhanced myeloid dendritic cell (DC) function. DCs from infected, but not uninfected, WT mice expressed significantly higher MHC class II and the co-stimulatory molecule CD80 compared to Tetherin KO DCs. Tetherin-associated DC activation during acute FV infection correlated with stronger NK cell responses. Furthermore, Tetherin+ DCs from FV-infected mice more strongly stimulated FV-specific CD4+ T cells ex vivo compared to Tetherin KO DCs. The results link the antiretroviral and immunomodulatory activity of Tetherin in vivo to improved DC activation and MHC class II antigen presentation. PMID:26846717

Complete genome sequences of two novel European clade bovine foamy viruses from Germany and Poland.

PubMed

Hechler, Torsten; Materniak, Magdalena; Kehl, Timo; Kuzmak, Jacek; Löchelt, Martin

2012-10-01

Bovine foamy virus (BFV), or bovine spumaretrovirus, is an infectious agent of cattle with no obvious disease association but high prevalence in its host. Here, we report two complete BFV sequences, BFV-Riems, isolated in 1978 in East Germany, and BFV100, isolated in 2005 in Poland. Both new BFV isolates share the overall genetic makeup of other foamy viruses (FV). Although isolated almost 25 years apart and propagated in either bovine (BFV-Riems) or nonbovine (BFV100) cells, both viruses are highly related, forming the European BFV clade. Despite clear differences, BFV-Riems and BFV100 are still very similar to BFV isolates from China and the United States, comprising the non-European BFV clade. The genomic sequences presented here confirm the concept of high sequence conservation across most of the FV genome. Analyses of cell culture-derived genomes reveal that proviral DNA may specifically lack introns in the env-bel coding region. The spacing of the splice sites in this region suggests that BFV has developed a novel mode to express a secretory but nonfunctional Env protein.

Complete Genome Sequences of Two Novel European Clade Bovine Foamy Viruses from Germany and Poland

PubMed Central

Hechler, Torsten; Materniak, Magdalena; Kehl, Timo; Kuzmak, Jacek

2012-01-01

Bovine foamy virus (BFV), or bovine spumaretrovirus, is an infectious agent of cattle with no obvious disease association but high prevalence in its host. Here, we report two complete BFV sequences, BFV-Riems, isolated in 1978 in East Germany, and BFV100, isolated in 2005 in Poland. Both new BFV isolates share the overall genetic makeup of other foamy viruses (FV). Although isolated almost 25 years apart and propagated in either bovine (BFV-Riems) or nonbovine (BFV100) cells, both viruses are highly related, forming the European BFV clade. Despite clear differences, BFV-Riems and BFV100 are still very similar to BFV isolates from China and the United States, comprising the non-European BFV clade. The genomic sequences presented here confirm the concept of high sequence conservation across most of the FV genome. Analyses of cell culture-derived genomes reveal that proviral DNA may specifically lack introns in the env-bel coding region. The spacing of the splice sites in this region suggests that BFV has developed a novel mode to express a secretory but nonfunctional Env protein. PMID:22966195

Effect of a creative-bonding intervention on Taiwanese nursing students' self-transcendence and attitudes toward elders.

PubMed

Chen, Shiue; Walsh, Sandra M

2009-04-01

Nursing students worldwide have little interest in caring for a growing elder population. The purpose of this study, based on self-transcendence theory, was to test the effectiveness of a creative-bonding intervention (CBI) to promote self-transcendence and positive attitudes towards elders in Taiwanese nursing students. A quasi-experimental design was used to compare a CBI group (n = 100) with a friendly visit (FV) control group (n = 94). ANCOVA results indicated that after the intervention, the CBI group had significantly more positive attitudes towards elders than the FV group. Nursing school faculty may want to promote art-making activities between students and elders to foster students' interest in elder care.

N-Myc Interactor Inhibits Prototype Foamy Virus by Sequestering Viral Tas Protein in the Cytoplasm

PubMed Central

Hu, Xiaomei; Yang, Wei; Liu, Ruikang; Geng, Yunqi; Qiao, Wentao

2014-01-01

ABSTRACT Foamy viruses (FVs) are complex retroviruses that establish lifelong persistent infection without evident pathology. However, the roles of cellular factors in FV latency are poorly understood. This study revealed that N-Myc interactor (Nmi) could inhibit the replication of prototype foamy virus (PFV). Overexpression of Nmi reduced PFV replication, whereas its depletion by small interfering RNA increased PFV replication. The Nmi-mediated impairment of PFV replication resulted from the diminished transactivation by PFV Tas of the viral long terminal repeat (LTR) and an internal promoter (IP). Nmi was determined to interact with Tas and abrogate its function by sequestration in the cytoplasm. In addition, human and bovine Nmi proteins were found to inhibit the replication of bovine foamy virus (BFV) and PFV. Together, these results indicate that Nmi inhibits both human and bovine FVs by interfering with the transactivation function of Tas and may have a role in the host defense against FV infection. IMPORTANCE From this study, we report that the N-Myc interactor (Nmi), an interferon-induced protein, can interact with the regulatory protein Tas of the prototype foamy virus and sequester it in the cytoplasm. The results of this study suggest that Nmi plays an important role in maintaining foamy virus latency and may reveal a new pathway in the interferon-mediated antiviral barrier against viruses. These findings are important for understanding virus-host relationships not only with FVs but potentially for other retroviruses as well. PMID:24719420

Safety and immunogenicity of a recombinant tetravalent dengue vaccine in 9-16 year olds: a randomized, controlled, phase II trial in Latin America.

PubMed

Villar, Luis Á; Rivera-Medina, Doris M; Arredondo-García, José Luis; Boaz, Mark; Starr-Spires, Linda; Thakur, Manoj; Zambrano, Betzana; Miranda, María C; Rivas, Enrique; Dayan, Gustavo H

2013-10-01

The dengue virus is a member of the Flavivirus (FV) genus, which also includes the yellow fever virus. Dengue disease is caused by any 1 of 4 dengue virus serotypes and is a serious public health concern in Latin America. This study evaluated the safety and immunogenicity of a candidate recombinant, live-attenuated, tetravalent dengue vaccine (CYD-TDV) in 9-16 year olds in Latin America. In this randomized, blinded, controlled study, volunteers received either 3 doses of CYD-TDV (n = 401) or placebo as first and second injection and tetanus/diphtheria/acellular pertussis vaccine as third injection (n = 199) at 0, 6 and 12 months. Adverse events were documented. Plaque reduction neutralization test antibody titers against the 4 CYD-TDV parental strains were measured before and 28 days after each dose. Seropositivity was defined as antibody titers ≥10 1/dil. The number of adverse reactions decreased after each successive CYD-TDV dose. After each CYD-TDV dose, antibody titers against all 4 serotypes were higher than baseline and respective predose titers. After the third dose of CYD-TDV, 100%, 98.6% and 93.4% of participants were seropositive for at least 2, at least 3 or all 4 serotypes, respectively. Higher antibody titers were observed in participants in the CYD-TDV group who were FV-seropositive at baseline compared with those who were FV-seronegative. CYD-TDV had a favorable safety profile and elicited antibody responses against all 4 dengue virus serotypes in 9-16 year olds in Latin America. These findings support the continued development of CYD-TDV.

Engineering intracellular active transport systems as in vivo biomolecular tools.

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

Bachand, George David; Carroll-Portillo, Amanda

2006-11-01

Active transport systems provide essential functions in terms of cell physiology and metastasis. These systems, however, are also co-opted by invading viruses, enabling directed transport of the virus to and from the cell's nucleus (i.e., the site of virus replication). Based on this concept, fundamentally new approaches for interrogating and manipulating the inner workings of living cells may be achievable by co-opting Nature's active transport systems as an in vivo biomolecular tool. The overall goal of this project was to investigate the ability to engineer kinesin-based transport systems for in vivo applications, specifically the collection of effector proteins (e.g., transcriptionalmore » regulators) within single cells. In the first part of this project, a chimeric fusion protein consisting of kinesin and a single chain variable fragment (scFv) of an antibody was successfully produced through a recombinant expression system. The kinesin-scFv retained both catalytic and antigenic functionality, enabling selective capture and transport of target antigens. The incorporation of a rabbit IgG-specific scFv into the kinesin established a generalized system for functionalizing kinesin with a wide range of target-selective antibodies raised in rabbits. The second objective was to develop methods of isolating the intact microtubule network from live cells as a platform for evaluating kinesin-based transport within the cytoskeletal architecture of a cell. Successful isolation of intact microtubule networks from two distinct cell types was demonstrated using glutaraldehyde and methanol fixation methods. This work provides a platform for inferring the ability of kinesin-scFv to function in vivo, and may also serve as a three-dimensional scaffold for evaluating and exploiting kinesin-based transport for nanotechnological applications. Overall, the technology developed in this project represents a first-step in engineering active transport system for in vivo applications. Further development could potentially enable selective capture of intracellular antigens, targeted delivery of therapeutic agents, or disruption of the transport systems and consequently the infection and pathogenesis cycle of biothreat agents.« less

Development of sugar chain-binding single-chain variable fragment antibody to adult T-cell leukemia cells using glyco-nanotechnology and phage display method.

PubMed

Muchima, Kaname; Todaka, Taro; Shinchi, Hiroyuki; Sato, Ayaka; Tazoe, Arisa; Aramaki, Rikiya; Kakitsubata, Yuhei; Yokoyama, Risa; Arima, Naomichi; Baba, Masanori; Wakao, Masahiro; Ito, Yuji; Suda, Yasuo

2018-04-01

Adult T-cell leukemia (ATL) is an intractable blood cancer caused by the infection of human T-cell leukemia virus type-1, and effective medical treatment is required. It is known that the structure and expression levels of cell surface sugar chains vary depending on cell states such as inflammation and cancer. Thus, it is expected that the antibody specific for ATL cell surface sugar chain would be an effective diagnostic tool and a strong candidate for the development of an anti-ATL drug. Here, we developed a stable sugar chain-binding single-chain variable fragment antibody (scFv) that can bind to ATL cells using a fibre-type Sugar Chip and phage display method. The fiber-type Sugar Chips were prepared using O-glycans released from ATL cell lines. The scFv-displaying phages derived from human B cells (diversity: 1.04 × 108) were then screened using the fiber-type Sugar Chips, and an O-glycan-binding scFv was obtained. The flow cytometry analysis revealed that the scFv predominantly bound to ATL cell lines. The sugar chain-binding properties of the scFv was evaluated by array-type Sugar Chip immobilized with a library of synthetic glycosaminoglycan disaccharide structures. Highly sulphated disaccharide structures were found to have high affinity to scFv.

Engineering a Single Chain Fv Antibody to αvβ6 Integrin using the Specificity-Determining Loop of a Foot-and-Mouth Disease Virus

PubMed Central

Kogelberg, Heide; Tolner, Berend; Thomas, Gareth J.; Di Cara, Danielle; Minogue, Shane; Ramesh, Bala; Sodha, Serena; Marsh, Dan; Lowdell, Mark W.; Meyer, Tim; Begent, Richard H.J.; Hart, Ian; Marshall, John F; Chester, Kerry

2010-01-01

Summary The αvβ6 integrin is a promising target for cancer therapy. Its expression is up-regulated de novo on many types of carcinoma where it may activate transforming growth factor-β1 and transforming growth factor-β3, interact with the specific extracellular matrix proteins and promote migration and invasion of tumour cells. The viral protein 1 (VP1) coat protein of the O1 British field strain serotype of foot-and-mouth disease virus is a high-affinity ligand for αvβ6, and we recently reported that a peptide derived from VP1 exhibited αvβ6-specific binding in vitro and in vivo. We hypothesized that this peptide could confer binding specificity of an antibody to αvβ6. A 17-mer peptide of VP1 was inserted into the complementary-determining region H3 loop of MFE-23, a murine single-chain Fv (scFv) antibody reactive with carcinoembryonic antigen (CEA). The resultant scFv (B6-1) bound to αvβ6 but retained residual reactivity with CEA. This was eliminated by point mutation (Y100bP) in the variable heavy-chain domain to create an scFv (B6-2) that was as structurally stable as MFE-23 and reacted specifically with αvβ6 but not α5β1, αvβ3, αvβ5, αvβ8 or CEA. B6-2 was internalized into αvβ6-expressing cells and inhibited αvβ6-dependent migration of carcinoma cells. B6-2 was subsequently humanized. The humanized form (B6-3) was obtained as a non-covalent dimer from secretion in Pichia pastoris (115 mg/l) and was a potent inhibitor of αvβ6-mediated cell adhesion. Thus, we have used a rational stepwise approach to create a humanized scFv with therapeutic potential to block αvβ6-mediated cancer cell invasion or to deliver and internalize toxins specifically to αvβ6-expressing tumours. PMID:18656482

Friend Leukemogenic Virus-neutralizing Antibody from Mouse Ascitic Fluid

PubMed Central

March, R. W.; Chirigos, M. A.; Hook, W. A.; Burka, B. L.

1967-01-01

Ascitic fluid antibody produced in C57/B1 mice immunized with Friend leukemogenic virus exhibited potent neutralizing activity. In vitro neutralization tests revealed that a mean neutralization index of 3.0 was achieved, and it was shown by sucrose gradient ultracentrifugation that this antibody resembled the 7S type. A mean yield of 6.7 ml of fluid per mouse per weekly paracentesis was obtained over an 8-week period. The ascitic fluid antibody to Friend virus was also active in vivo. Mice given antibody 3, 5, 7, and 9 days after infection with Friend virus did not develop the splenomegaly characteristic of Friend disease. PMID:16349755

Complete sequence determination of a novel reptile iridovirus isolated from soft-shelled turtle and evolutionary analysis of Iridoviridae

PubMed Central

Huang, Youhua; Huang, Xiaohong; Liu, Hong; Gong, Jie; Ouyang, Zhengliang; Cui, Huachun; Cao, Jianhao; Zhao, Yingtao; Wang, Xiujie; Jiang, Yulin; Qin, Qiwei

2009-01-01

Background Soft-shelled turtle iridovirus (STIV) is the causative agent of severe systemic diseases in cultured soft-shelled turtles (Trionyx sinensis). To our knowledge, the only molecular information available on STIV mainly concerns the highly conserved STIV major capsid protein. The complete sequence of the STIV genome is not yet available. Therefore, determining the genome sequence of STIV and providing a detailed bioinformatic analysis of its genome content and evolution status will facilitate further understanding of the taxonomic elements of STIV and the molecular mechanisms of reptile iridovirus pathogenesis. Results We determined the complete nucleotide sequence of the STIV genome using 454 Life Science sequencing technology. The STIV genome is 105 890 bp in length with a base composition of 55.1% G+C. Computer assisted analysis revealed that the STIV genome contains 105 potential open reading frames (ORFs), which encode polypeptides ranging from 40 to 1,294 amino acids and 20 microRNA candidates. Among the putative proteins, 20 share homology with the ancestral proteins of the nuclear and cytoplasmic large DNA viruses (NCLDVs). Comparative genomic analysis showed that STIV has the highest degree of sequence conservation and a colinear arrangement of genes with frog virus 3 (FV3), followed by Tiger frog virus (TFV), Ambystoma tigrinum virus (ATV), Singapore grouper iridovirus (SGIV), Grouper iridovirus (GIV) and other iridovirus isolates. Phylogenetic analysis based on conserved core genes and complete genome sequence of STIV with other virus genomes was performed. Moreover, analysis of the gene gain-and-loss events in the family Iridoviridae suggested that the genes encoded by iridoviruses have evolved for favoring adaptation to different natural host species. Conclusion This study has provided the complete genome sequence of STIV. Phylogenetic analysis suggested that STIV and FV3 are strains of the same viral species belonging to the Ranavirus genus in the Iridoviridae family. Given virus-host co-evolution and the phylogenetic relationship among vertebrates from fish to reptiles, we propose that iridovirus might transmit between reptiles and amphibians and that STIV and FV3 are strains of the same viral species in the Ranavirus genus. PMID:19439104

Development of anti-bovine IgA single chain variable fragment and its application in diagnosis of foot-and-mouth disease

PubMed Central

Sridevi, N. V.; Shukra, A. M.; Neelakantam, B.; Anilkumar, J.; Madhanmohan, M.; Rajan, S.; Dev Chandran

2014-01-01

Recombinant antibody fragments like single chain variable fragments (scFvs) represent an attractive yet powerful alternative to immunoglobulins and hold great potential in the development of clinical diagnostic/therapeutic reagents. Structurally, scFvs are the smallest antibody fragments capable of retaining the antigen-binding capacity of whole antibodies and are composed of an immunoglobulin (Ig) variable light (VL) and variable heavy (VH) chain joined by a flexible polypeptide linker. In the present study, we constructed a scFv against bovine IgA from a hybridoma cell line IL-A71 that secretes a monoclonal antibody against bovine IgA using recombinant DNA technology. The scFv was expressed in Escherichia coli and purified using immobilized metal affinity chromatography (IMAC). The binding activity and specificity of the scFv was established by its non-reactivity toward other classes of immunoglobulins as determined by enzyme-linked immunosorbent assay (ELISA) and immunoblot analysis. Kinetic measurement of the scFv indicated that the recombinant antibody fragment had an affinity in picomolar range toward purified IgA. Furthermore, the scFv was used to develop a sensitive ELISA for the detection of foot and mouth disease virus (FMDV) carrier animals. PMID:24678404

Nuclear factors that bind to the enhancer region of nondefective Friend murine leukemia virus.

PubMed Central

Manley, N R; O'Connell, M A; Sharp, P A; Hopkins, N

1989-01-01

Nondefective Friend murine leukemia virus (MuLV) causes erythroleukemia when injected into newborn NFS mice, while Moloney MuLV causes T-cell lymphoma. Exchange of the Friend virus enhancer region, a sequence of about 180 nucleotides including the direct repeat and a short 3'-adjacent segment, for the corresponding region in Moloney MuLV confers the ability to cause erythroid disease on Moloney MuLV. We have used the electrophoretic mobility shift assay and methylation interference analysis to identify cellular factors which bind to the Friend virus enhancer region and compared these with factors, previously identified, that bind to the Moloney virus direct repeat (N. A. Speck and D. Baltimore, Mol. Cell. Biol. 7:1101-1110, 1987). We identified five binding sites for sequence-specific DNA-binding proteins in the Friend virus enhancer region. While some binding sites are present in both the Moloney and Friend virus enhancers, both viruses contain unique sites not present in the other. Although none of the factors identified in this report which bind to these unique sites are present exclusively in T cells or erythroid cells, they bind to three regions of the enhancer shown by genetic analysis to encode disease specificity and thus are candidates to mediate the tissue-specific expression and distinct disease specificities encoded by these virus enhancer elements. Images PMID:2778872

Deciphering the behaviour manipulation imposed by a virus on its parasitoid host: insights from a dual transcriptomic approach.

PubMed

Varaldi, J; Lepetit, D

2018-05-28

Behaviour manipulation imposed by parasites is a fascinating phenomenon but our understanding is still very limited. We studied the interaction between a virus and the parasitic wasp Leptopilina boulardi that attacks Drosophila larvae. Wasps usually refrain to lay eggs into already parasitized hosts (superparasitism avoidance). On the contrary, females infected by the Leptopilina boulardi Filamentous Virus (LbFV) are much more incline to superparasitize. Interestingly, the host-sharing induced by this behaviour modification leads to the horizontal transmission of the virus, thus increasing its fitness at the expense of that of the wasp. To better understand the mechanisms underlying this behaviour manipulation, we studied by RNA sequencing the meta-transcriptome of LbFV and the parasitic wasp both in the abdomen and in the head. We found that the abundance of viral transcripts was independent of the wasp strain but strongly differed between tissues. Based on the tissue pattern of expression, we identified a set of 20 viral genes putatively involved in the manipulation process. In addition, we identified a set of wasp genes deregulated in the presence of the virus either in the abdomen or in the head, including genes with annotations suggesting involvement in behaviour (i.e. Potassium-channel protein). This dataset gives new insights into the behaviour manipulation and on the genetic basis of superparasitism in parasitoids.

A baculovirus-mediated strategy for full-length plant virus coat protein expression and purification.

PubMed

Ardisson-Araújo, Daniel Mendes Pereira; Rocha, Juliana Ribeiro; da Costa, Márcio Hedil Oliveira; Bocca, Anamélia Lorenzetti; Dusi, André Nepomuceno; de Oliveira Resende, Renato; Ribeiro, Bergmann Morais

2013-08-15

Garlic production is severely affected by virus infection, causing a decrease in productivity and quality. There are no virus-free cultivars and garlic-infecting viruses are difficult to purify, which make specific antibody production very laborious. Since high quality antisera against plant viruses are important tools for serological detection, we have developed a method to express and purify full-length plant virus coat proteins using baculovirus expression system and insects as bioreactors. In this work, we have fused the full-length coat protein (cp) gene from the Garlic Mite-borne Filamentous Virus (GarMbFV) to the 3'-end of the Polyhedrin (polh) gene of the baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV). The recombinant baculovirus was amplified in insect cell culture and the virus was used to infect Spodoptera frugiperda larvae. Thus, the recombinant fused protein was easily purified from insect cadavers using sucrose gradient centrifugation and analyzed by Western Blotting. Interestingly, amorphous crystals were produced in the cytoplasm of cells infected with the recombinant virus containing the chimeric-protein gene but not in cells infected with the wild type and recombinant virus containing the hexa histidine tagged Polh. Moreover, the chimeric protein was used to immunize rats and generate antibodies against the target protein. The antiserum produced was able to detect plants infected with GarMbFV, which had been initially confirmed by RT-PCR. The expression of a plant virus full-length coat protein fused to the baculovirus Polyhedrin in recombinant baculovirus-infected insects was shown to produce high amounts of the recombinant protein which was easily purified and efficiently used to generate specific antibodies. Therefore, this strategy can potentially be used for the development of plant virus diagnostic kits for those viruses that are difficult to purify, are present in low titers or are present in mix infection in their plant hosts.

Erythroid colony induction without erythropoietin by Friend leukemia virus in vitro.

PubMed

Clarke, B J; Axelrad, A A; Shreeve, M M; McLeod, D L

1975-09-01

Erythroid colonies could be produced without the addition of erythropeietin in plasma cultures seeded with bone marrow cells from normal C3Hf/Bi mice by exposure of the cells in vitro to medium from a cell line (IS) that continuously produces Friend leukemia virus in culture. The activity in the culture medium was viral rather than erythropoietin-like, since it was sedimentable by high-speed centrifugation and heat labile. Erythroid colonies did not develop when the bone marrow cells exposed to virus-containing medium were from mice genetically resistant to Friend virus. IS culture medium contained both Friend spleen focus-forming and XC-plaque-forming activities. No erythroid colonies were induced when genetically sensitive cells were exposed to a preparation from which the spleen focus-forming activity had been removed, but which contained XC plaque-forming activity in high concentration. Thus the spleen focus-forming component of Friend virus appeared to be responsible for inducing erythroid colony formation without erythropoietin in vitro. Some erythroid colonies were also found in control cultures to which neither virus nor erythropoietin had been added. Reduction in the concentration of fetal calf serum in the culture medium substantially decreased the number of these colonies but had only a minor effect on the number of virus-induced colonies. The number of erythroid colonies produced after 2 days of culture without erythropoietin or fetal calf serum was approximately proportional to the titer of Friend spleen focus-forming virus to whcih the bone marrow cells had been exposed. This system should prove useful for investigation in vitro of Friend virus--host cell interactions which lead to erythropoietin-independent erythropoiesis.

Friend and Moloney murine leukemia viruses specifically recombine with different endogenous retroviral sequences to generate mink cell focus-forming viruses.

PubMed

Evans, L H; Cloyd, M W

1985-01-01

A group of mink cell focus-forming (MCF) viruses was derived by inoculation of NFS/N mice with Moloney murine leukemia virus (Mo-MuLV 1387) and was compared to a similarly derived group of MCF viruses from mice inoculated with Friend MuLV (Fr-MuLV 57). Antigenic analyses using monoclonal antibodies specific for MCF virus and xenotropic MuLV envelope proteins and genomic structural analyses by RNase T1-resistant oligonucleotide finger-printing indicated that the Moloney and Friend MCF viruses arose by recombination of the respective ecotropic MuLVs with different endogenous retrovirus sequences of NFS mice.

Selective anti-herpesvirus agents.

PubMed

De Clercq, Erik

2013-01-23

This review article focuses on the anti-herpesvirus agents effective against herpes simplex virus, varicella-zoster virus and cytomegalovirus, which have either been licensed for clinical use (idoxuridine, trifluridine, brivudin, acyclovir, valaciclovir, valganciclovir, famciclovir and foscarnet) or are under clinical development (CMX001 [the hexadecyloxypropyl prodrug of cidofovir], the helicase-primase inhibitor BAY 57-1293 [now referred to as AIC316], FV-100 [the valine ester of Cf 1743] and the terminase inhibitor letermovir [AIC246]).

Antiviral Immunotoxin Against Bovine herpesvirus-1: Targeted Inhibition of Viral Replication and Apoptosis of Infected Cell

PubMed Central

Xu, Jian; Li, Xiaoyang; Jiang, Bo; Feng, Xiaoyu; Wu, Jing; Cai, Yunhong; Zhang, Xixi; Huang, Xiufen; Sealy, Joshua E.; Iqbal, Munir; Li, Yongqing

2018-01-01

Bovine herpesvirus 1 (BoHV-1) is a highly contagious viral pathogen which causes infectious bovine rhinotracheitis in cattle worldwide. Currently, there is no antiviral prophylactic treatment available capable of mitigating the disease impact and facilitating recovery from latent infection. In this study, we have engineered a novel recombinant anti-BoHV-1 immunotoxin construct termed “BoScFv-PE38” that consists of a single-chain monoclonal antibody fragment (scFv) fused with an active domain of Pseudomonas exotoxin A as a toxic effector (PE38). The recombinant BoScFv-PE38 immunotoxin expressed in a prokaryotic expression system has specific binding affinity for BoHV-1 glycoprotein D (gD) with a dissociation constant (Kd) of 12.81 nM and for BoHV-1 virus particles with a Kd value of 97.63 nM. We demonstrate that the recombinant BoScFv-PE38 is internalized into MDBK cell compartments that inhibit BoHV-1 replication with a half-maximal inhibitory concentration (IC50) of 4.95 ± 0.33 nM and a selective index (SI) of 456 ± 31. Furthermore, the BoScFv-PE38 exerted a cytotoxic effect through the induction of ATP and ammonia, leading to apoptosis of BoHV-1-infected cells and the inhibition of BoHV-1 replication in MDBK cells. Collectively, we show that BoScFv-PE38 can potentially be employed as a therapeutic agent for the treatment of BoHV-1 infection. PMID:29670605

Bovine single chain Fv antibody inhibits bovine herpesvirus-1 infectivity by targeting viral glycoprotein D.

PubMed

Xu, Jian; Wu, Jing; Jiang, Bo; He, Houjun; Zhang, Xixi; Li, Xiaoyang; Yang, Dawei; Huang, Xiufen; Sealy, Joshua E; Iqbal, Munir; Li, Yongqing

2017-12-01

Glycoprotein D (gD) of bovine herpesvirus-1 (BoHV-1) is essential for attachment and penetration of cells during infection and is a major target for neutralizing antibodies during an adaptive immune response. Currently there are no recombinant antibodies capable of binding gD epitopes for use in treating BoHV-1 infection. In this study, a bovine scFv gene derived from a hybridoma secreting monoclonal antibodies (McAbs) against the amino acid motif MEESKGYEPP of gD was expressed in E. coli. Molecular modeling, western blot and ELISA analysis showed that this scFv had a high affinity for BoHV-1 gD, with a Kd of 161.2 ± 37.58 nM and for whole BoHV-1 virus, with a Kd of 67.44 ± 16.99 nM. In addition, this scFv displayed a high affinity for BoHV-1 antigen in an ELISA and competed with BoHV-1 anti-serum in a competitive ELISA. Immunofluorescence assay (IFA) and laser confocal microscopy showed that this scFv could efficiently bind to and be internalized by BoHV-1 infected Madin-Darby bovine kidney (MDBK) cells. Importantly, this scFv was shown to inhibit BoHV-1 infectivity and to reduce the number of viral plaques by blocking viral attachment to MDBK cells. Our study suggests that this bovine single-chain antibody could be developed for use as a diagnostic and therapeutic agent against BoHV-1 infection in cattle.

The Envelope Glycoprotein of Friend Spleen Focus-Forming Virus Covalently Interacts with and Constitutively Activates a Truncated Form of the Receptor Tyrosine Kinase Stk

PubMed Central

Nishigaki, Kazuo; Thompson, Delores; Hanson, Charlotte; Yugawa, Takashi; Ruscetti, Sandra

2001-01-01

The Friend spleen focus-forming virus (SFFV) encodes a unique envelope glycoprotein, gp55, which allows erythroid cells to proliferate and differentiate in the absence of erythropoietin (Epo). SFFV gp55 has been shown to interact with the Epo receptor complex, causing constitutive activation of various signal-transducing molecules. When injected into adult mice, SFFV induces a rapid erythroleukemia, with susceptibility being determined by the host gene Fv-2, which was recently shown to be identical to the gene encoding the receptor tyrosine kinase Stk/Ron. Susceptible, but not resistant, mice encode not only full-length Stk but also a truncated form of the kinase, sf-Stk, which may mediate the biological effects of SFFV infection. To determine whether expression of SFFV gp55 leads to the activation of sf-Stk, we expressed sf-Stk, with or without SFFV gp55, in hematopoietic cells expressing the Epo receptor. Our data indicate that sf-Stk interacts with SFFV gp55 as well as gp55P, the biologically active form of the viral glycoprotein, forming disulfide-linked complexes. This covalent interaction, as well as noncovalent interactions with SFFV gp55, results in constitutive tyrosine phosphorylation of sf-Stk and its association with multiple tyrosine-phosphorylated signal-transducing molecules. In contrast, neither Epo stimulation in the absence of SFFV gp55 expression nor expression of a mutant of SFFV that cannot interact with sf-Stk was able to induce tyrosine phosphorylation of sf-Stk or its association with any signal-transducing molecules. Covalent interaction of sf-Stk with SFFV gp55 and constitutive tyrosine phosphorylation of sf-Stk can also be detected in an erythroleukemia cell line derived from an SFFV-infected mouse. Our results suggest that SFFV gp55 may mediate its biological effects in vivo by interacting with and activating a truncated form of the receptor tyrosine kinase Stk. PMID:11483734

The Acaricidal Activity of Venom from the Jellyfish Nemopilema nomurai against the Carmine Spider Mite Tetranychus cinnabarinus

PubMed Central

Yu, Huahua; Yue, Yang; Dong, Xiangli; Li, Rongfeng; Li, Pengcheng

2016-01-01

The carmine spider mite Tetranychus cinnabarinus (T. cinnabarinus) is a common polyphagous pest that attacks crops, vegetables, flowers, and so on. It is necessary to find lead compounds for developing novel, powerful, and environmentally-friendly acaricides as an alternative approach to controlling the carmine spider mite because of the serious resistance and residual agrochemicals in the environment. In addition, the study on the acaricidal activities of marine bioactive substances is comparatively deficient. In the present study, the acaricidal activity of venom (NnFV) from the jellyfish Nemopilema nomurai against the carmine spider mite T. cinnabarinus was determined for the first time. The venom had contact toxicity, and the 24-h LC50-value was 29.1 μg/mL. The mite body wall was affected by the venom, with the mite body having no luster and being seriously shrunken after 24 h. T. cinnabarinus was a potential target pest of NnFV, which had potential as a type of natural bioacaricide. The repellent activity and systemic toxicity of the venom against T. cinnabarinus were also studied. However, NnFV had no repellent activity and systemic toxicity against T. cinnabarinus. PMID:27294957

Bifunctional Anti-Huntingtin Proteasome-Directed Intrabodies Mediate Efficient Degradation of Mutant Huntingtin Exon 1 Protein Fragments

PubMed Central

Butler, David C.; Messer, Anne

2011-01-01

Huntington's disease (HD) is a fatal autosomal dominant neurodegenerative disorder caused by a trinucleotide (CAG)n repeat expansion in the coding sequence of the huntingtin gene, and an expanded polyglutamine (>37Q) tract in the protein. This results in misfolding and accumulation of huntingtin protein (htt), formation of neuronal intranuclear and cytoplasmic inclusions, and neuronal dysfunction/degeneration. Single-chain Fv antibodies (scFvs), expressed as intrabodies that bind htt and prevent aggregation, show promise as immunotherapeutics for HD. Intrastriatal delivery of anti-N-terminal htt scFv-C4 using an adeno-associated virus vector (AAV2/1) significantly reduces the size and number of aggregates in HDR6/1 transgenic mice; however, this protective effect diminishes with age and time after injection. We therefore explored enhancing intrabody efficacy via fusions to heterologous functional domains. Proteins containing a PEST motif are often targeted for proteasomal degradation and generally have a short half life. In ST14A cells, fusion of the C-terminal PEST region of mouse ornithine decarboxylase (mODC) to scFv-C4 reduces htt exon 1 protein fragments with 72 glutamine repeats (httex1-72Q) by ∼80–90% when compared to scFv-C4 alone. Proteasomal targeting was verified by either scrambling the mODC-PEST motif, or via proteasomal inhibition with epoxomicin. For these constructs, the proteasomal degradation of the scFv intrabody proteins themselves was reduced<25% by the addition of the mODC-PEST motif, with or without antigens. The remaining intrabody levels were amply sufficient to target N-terminal httex1-72Q protein fragment turnover. Critically, scFv-C4-PEST prevents aggregation and toxicity of httex1-72Q fragments at significantly lower doses than scFv-C4. Fusion of the mODC-PEST motif to intrabodies is a valuable general approach to specifically target toxic antigens to the proteasome for degradation. PMID:22216210

Production of antibody labeled gold nanoparticles for influenza virus H5N1 diagnosis kit development

NASA Astrophysics Data System (ADS)

Pham, Van Dong; Hoang, Ha; Hoang Phan, Trong; Conrad, Udo; Chu, Hoang Ha

2012-12-01

Preparation of colloidal gold conjugated antibodies specific for influenza A/H5N1 and its use in developing a virus A/H5N1 rapid diagnostic kit is presented. Colloidal gold nanoparticles (AuNPs) were prepared through citrate reduction. Single chain antibodies specific to H5N1 (scFv7 and scFv24) were produced using pTI2 + vector and E. coli strain HB2151. These antibodies were purified by affinity chromatography technique employing HiTrap Chelating HP columns pre-charged with Ni2 + . The method for preparation of antibody-colloidal gold conjugate was based on electrostatic force binding antibody with colloidal gold. The effect of factors such as pH and concentration of antibody has been quantitatively analyzed using spectroscopic methods after adding 1 wt% NaCl which induced AuNP aggregation. The morphological study by scanning electron microscopy (SEM) showed that the average size of the spherical AuNPs was 23 nm with uniform sizes. The spectroscopic properties of colloidal AuNPs showed the typical surface plasmon resonance band at 523 nm in UV-visible spectrum. The optimal pH of conjugated colloidal gold was found between 8.0 and 10.0. The activity of synthesized antibody labeled AuNPs for detection of H5N1 flu virus was checked by dot blot immunological method. The results confirmed the ability in detection of the A/H5N1 virus of the prepared antibody labeled gold particles and opened up the possibility of using them in manufacturing rapid detection kit for this virus.

Structure of a Spumaretrovirus Gag Central Domain Reveals an Ancient Retroviral Capsid

PubMed Central

Dutta, Moumita; Pollard, Dominic J.; Goldstone, David C.; Ramos, Andres; Müllers, Erik; Stirnnagel, Kristin; Stanke, Nicole; Lindemann, Dirk; Taylor, William R.; Rosenthal, Peter B.

2016-01-01

The Spumaretrovirinae, or foamy viruses (FVs) are complex retroviruses that infect many species of monkey and ape. Despite little sequence homology, FV and orthoretroviral Gag proteins perform equivalent functions, including genome packaging, virion assembly, trafficking and membrane targeting. However, there is a paucity of structural information for FVs and it is unclear how disparate FV and orthoretroviral Gag molecules share the same function. To probe the functional overlap of FV and orthoretroviral Gag we have determined the structure of a central region of Gag from the Prototype FV (PFV). The structure comprises two all α-helical domains NtDCEN and CtDCEN that although they have no sequence similarity, we show they share the same core fold as the N- (NtDCA) and C-terminal domains (CtDCA) of archetypal orthoretroviral capsid protein (CA). Moreover, structural comparisons with orthoretroviral CA align PFV NtDCEN and CtDCEN with NtDCA and CtDCA respectively. Further in vitro and functional virological assays reveal that residues making inter-domain NtDCEN—CtDCEN interactions are required for PFV capsid assembly and that intact capsid is required for PFV reverse transcription. These data provide the first information that relates the Gag proteins of Spuma and Orthoretrovirinae and suggests a common ancestor for both lineages containing an ancient CA fold. PMID:27829070

Foamy virus–mediated gene transfer to canine repopulating cells

PubMed Central

Kiem, Hans-Peter; Allen, James; Trobridge, Grant; Olson, Erik; Keyser, Kirsten; Peterson, Laura; Russell, David W.

2007-01-01

Foamy virus (FV) vectors are particularly attractive gene-transfer vectors for stem-cell gene therapy because they form a stable transduction intermediate in quiescent cells and can efficiently transduce hematopoietic stem cells. Here, we studied the use of FV vectors to transduce long-term hematopoietic repopulating cells in the dog, a clinically relevant large animal model. Mobilized canine peripheral blood (PB) CD34+ cells were transduced with an enhanced green fluorescent protein (EGFP)–expressing FV vector in an 18-hour transduction protocol. All 3 dogs studied had rapid neutrophil engraftment to greater than 500/μL with a median of 10 days. Transgene expression was detected in all cell lineages (B cells, T cells, granulocytes, red blood cells, and platelets), indicating multilineage engraftment of transduced cells. Up to 19% of blood cells were EGFP+, and this was confirmed at the DNA level by real-time polymerase chain reaction (PCR) and Southern blot analysis. These transduction rates were higher than the best results we obtained previously with lentiviral vectors in a similar transduction protocol. Integration site analysis also demonstrated polyclonal repopulation and the transduction of multipotential hematopoietic repopulating cells. These data suggest that FV vectors should be useful for stem-cell gene therapy, particularly for applications in which short transduction protocols are critical. PMID:16968897

Corexit 9500 inactivates two enveloped viruses of aquatic animals but enhances the infectivity of a nonenveloped fish virus.

PubMed

Pham, P H; Huang, Y J; Chen, C; Bols, N C

2014-02-01

The effects of Corexit 9500, a dispersant used to clean up oil spills, on invertebrates, lower vertebrates, birds, and human health have been examined, but there is a significant lack of study of the effect of this dispersant on aquatic viruses. In this study, the effects of Corexit 9500 on four aquatic viruses of differing structural composition were examined. Corexit 9500 reduced the titer of the enveloped viral hemorrhagic septicemia virus (VHSV) at all concentrations (10% to 0.001%) examined. The titer of frog virus 3 (FV3), a virus with both enveloped and nonenveloped virions, was reduced only at the high Corexit 9500 concentrations (10% to 0.1%). Corexit 9500 was unable to reduce the titer of nonenveloped infectious pancreatic necrosis virus (IPNV) but enhanced the titer of chum salmon reovirus (CSV) by 2 to 4 logs. With the ability to inactivate enveloped viruses and possibly enhance some nonenveloped viruses, Corexit 9500 has the potential to alter the aquatic virosphere.

Experimental Transmission of Frog Virus 3-Like Ranavirus in Juvenile Chelonians at Two Temperatures.

PubMed

Allender, Matthew C; Barthel, Ashley C; Rayl, Jeremy M; Terio, Karen A

2018-06-07

The pathogenicity of frog virus 3 (FV3)-like ranavirus varies in adult chelonian species at different environmental temperatures, but differences in pathogenicity at different temperatures has yet to be determined in juveniles. Our objective was to determine the susceptibility to FV3-like ranavirus in four species of juvenile chelonians: red-eared sliders (RES; Trachemys scripta elegans), Mississippi map turtles ( Graptemys pseudogeographica kohnii), false map turtles (FMT; Graptemys pseudogeographica), and eastern river cooters ( Pseudemys concinna concinna) at two environmental temperatures. Two simultaneous trials ( n=8 treatment and n=4 controls of each species) were conducted in separate temperature-controlled rooms with animals maintained at 22 C or 27 C. All of the inoculated animals of each species at each temperature died, but no mortality was observed in control animals. Median survival times varied between 8 d and 11 d, based on species and temperature, with RES in the 27 C trial surviving the shortest time and the FMT in the 22 C trial surviving the longest. Combining all species, turtles in the 27 C trial survived for fewer days than those housed at 22 C, despite all turtles in both trials having similar viral copies detected in postmortem tissues. Lesions in inoculated turtles resembled those noted in natural and experimental FV3-like ranavirus infections and included vasculitis, thrombosis, hemorrhage in multiple organs, renal tubular necrosis, and hepatic necrosis. Myositis was not present in any juvenile, infected turtles in this study. This study confirmed that juvenile chelonians have a high susceptibility to ranaviral disease.

Rapid immune reconstitution of SCID-X1 canines after G-CSF/AMD3100 mobilization and in vivo gene therapy

PubMed Central

Humbert, Olivier; Chan, Frieda; Rajawat, Yogendra S.; Torgerson, Troy R.; Burtner, Christopher R.; Hubbard, Nicholas W.; Humphrys, Daniel; Norgaard, Zachary K.; O’Donnell, Patricia; Adair, Jennifer E.; Trobridge, Grant D.; Scharenberg, Andrew M.; Felsburg, Peter J.; Rawlings, David J.

2018-01-01

Hematopoietic stem-cell gene therapy is a promising treatment of X-linked severe combined immunodeficiency disease (SCID-X1), but currently, it requires recipient conditioning, extensive cell manipulation, and sophisticated facilities. With these limitations in mind, we explored a simpler therapeutic approach to SCID-X1 treatment by direct IV administration of foamy virus (FV) vectors in the canine model. FV vectors were used because they have a favorable integration site profile and are resistant to serum inactivation. Here, we show improved efficacy of our in vivo gene therapy platform by mobilization with granulocyte colony-stimulating factor (G-CSF) and AMD3100 before injection of an optimized FV vector incorporating the human phosphoglycerate kinase enhancerless promoter. G-CSF/AMD3100 mobilization before FV vector delivery accelerated kinetics of CD3+ lymphocyte recovery, promoted thymopoiesis, and increased immune clonal diversity. Gene-corrected T lymphocytes exhibited a normal CD4:CD8 ratio and a broad T-cell receptor repertoire and showed restored γC-dependent signaling function. Treated animals showed normal primary and secondary antibody responses to bacteriophage immunization and evidence for immunoglobulin class switching. These results demonstrate safety and efficacy of an accessible, portable, and translatable platform with no conditioning regimen for the treatment of SCID-X1 and other genetic diseases. PMID:29720491

FV-100: the most potent and selective anti-varicella zoster virus agent reported to date.

PubMed

Migliore, Marco

2010-01-05

Bicyclic aryl furano pyrimidines represent the most potent anti-varicella zoster virus (VZV) agents reported to date. Lead compounds have 50% effective concentration (EC(50)) values in vitro that are in the subnanomolar range and selectivity index values that exceed 1 million. They have an absolute requirement for VZV thymidine kinase and most likely act as their phosphate forms. Some structural modification (such as aryl substitution in the base moiety) is tolerated, whereas little sugar modification is acceptable. The Cf1743 compound has proved to be significantly more potent than all reference anti-VZV compounds, as measured either by inhibition of infectious virus particles and/or viral DNA production; however, the high lipophilicity and very low water solubility of this compound gives poor oral bioavailability (<14%). Use of the modified cyclodextrin captisol and the synthesis of the 5'-monophosphate prodrug of Cf1743 has significantly improved water solubility, but does not give any enhancement in oral bioavailability. By contrast, the synthesis of the ether series does not give any further improvement in terms of solubility. The most promising prodrug to emerge to date is the hydrochloric salt of the 5'-valyl-ester, designated as FV-100. Its uptake into cells has been studied using fluorescent microscopy and biological assays, which have indicated that the compound is efficiently taken up by the cells after a short period of incubation.

Antivirion Effects of Streptovaricin Complex Against Friend Virus

PubMed Central

Horoszewicz, Julius S.; Leong, Susan S.; Byrd, Daniel M.; Carter, William A.

1974-01-01

The in vitro antivirion activities of five different streptovaricin complex lots against the polycythemic strain of the Friend virus were evaluated. The assay system was based on the inhibition of the Friend virus-induced spleen foci. The virus inactivation process was shown to be susceptible to variation in temperature, pH, and time. The antivirion activity and the acute toxicity for mice, as well as the optical properties of these streptovaricin complexes, do not co-vary; this suggests that their biological activities are not associated with a single molecular structure. In addition, the antivirion activity of the five preparations of streptovaricin complex differs about 30-fold, indicating that this activity does not reside in a major component of the complex. PMID:15825311

A novel small animal model to study the replication of simian foamy virus in vivo.

PubMed

Blochmann, Rico; Curths, Christoph; Coulibaly, Cheick; Cichutek, Klaus; Kurth, Reinhard; Norley, Stephen; Bannert, Norbert; Fiebig, Uwe

2014-01-05

Preclinical evaluation in a small animal model would help the development of gene therapies and vaccines based on foamy virus vectors. The establishment of persistent, non-pathogenic infection with the prototype foamy virus in mice and rabbits has been described previously. To extend this spectrum of available animal models, hamsters were inoculated with infectious cell supernatant or bioballistically with a foamy virus plasmid. In addition, a novel foamy virus from a rhesus macaque was isolated and characterised genetically. Hamsters and mice were infected with this new SFVmac isolate to evaluate whether hamsters are also susceptible to infection. Both hamsters and mice developed humoral responses to either virus subtype. Virus integration and replication in different animal tissues were analysed by PCR and co-cultivation. The results strongly indicate establishment of a persistent infection in hamsters. These studies provide a further small animal model for studying FV-based vectors in addition to the established models. © 2013 Elsevier Inc. All rights reserved.

Viral Membrane Fusion and Nucleocapsid Delivery into the Cytoplasm are Distinct Events in Some Flaviviruses

PubMed Central

Nour, Adel M.; Li, Yue; Wolenski, Joseph; Modis, Yorgo

2013-01-01

Flaviviruses deliver their genome into the cell by fusing the viral lipid membrane to an endosomal membrane. The sequence and kinetics of the steps required for nucleocapsid delivery into the cytoplasm remain unclear. Here we dissect the cell entry pathway of virions and virus-like particles from two flaviviruses using single-particle tracking in live cells, a biochemical membrane fusion assay and virus infectivity assays. We show that the virus particles fuse with a small endosomal compartment in which the nucleocapsid remains trapped for several minutes. Endosomal maturation inhibitors inhibit infectivity but not membrane fusion. We propose a flavivirus cell entry mechanism in which the virus particles fuse preferentially with small endosomal carrier vesicles and depend on back-fusion of the vesicles with the late endosomal membrane to deliver the nucleocapsid into the cytoplasm. Virus entry modulates intracellular calcium release and phosphatidylinositol-3-phosphate kinase signaling. Moreover, the broadly cross-reactive therapeutic antibody scFv11 binds to virus-like particles and inhibits fusion. PMID:24039574

Clinical signs, pathology and dose-dependent survival of adult wood frogs, Rana sylvatica, inoculated orally with frog virus 3 Ranavirus sp., Iridoviridae.

PubMed

Forzn, Mara J; Jones, Kathleen M; Vanderstichel, Raphal V; Wood, John; Kibenge, Frederick S B; Kuiken, Thijs; Wirth, Wytamma; Ariel, Ellen; Daoust, Pierre-Yves

2015-05-01

Amphibian populations suffer massive mortalities from infection with frog virus 3 FV3, genus Ranavirus, family Iridoviridae, a pathogen also involved in mortalities of fish and reptiles. Experimental oral infection with FV3 in captive-raised adult wood frogs, Rana sylvatica Lithobates sylvaticus, was performed as the first step in establishing a native North American animal model of ranaviral disease to study pathogenesis and host response. Oral dosing was successful LD50 was 10(2.93 2.423.44) p.f.u. for frogs averaging 35mm in length. Onset of clinical signs occurred 614days post-infection p.i. median 11 days p.i. and time to death was 1014 days p.i. median 12 days p.i.. Each tenfold increase in virus dose increased the odds of dying by 23-fold and accelerated onset of clinical signs and death by approximately 15. Ranavirus DNA was demonstrated in skin and liver of all frogs that died or were euthanized because of severe clinical signs. Shedding of virus occurred in faeces 710 days p.i. 34.5days before death and skin sheds 10 days p.i. 01.5days before death of some frogs dead from infection. Most common lesions were dermal erosion and haemorrhages haematopoietic necrosis in bone marrow, kidney, spleen and liver and necrosis in renal glomeruli, tongue, gastrointestinal tract and urinary bladder mucosa. Presence of ranavirus in lesions was confirmed by immunohistochemistry. Intracytoplasmic inclusion bodies probably viral were present in the bone marrow and the epithelia of the oral cavity, gastrointestinal tract, renal tubules and urinary bladder. Our work describes a ranaviruswood frog model and provides estimates that can be incorporated into ranavirus disease ecology models. © 2015 The Authors.

Factor V Has Anticoagulant Activity in Plasma in the Presence of TFPIα: Difference between FV1 and FV2.

PubMed

van Doorn, Peter; Rosing, Jan; Duckers, Connie; Hackeng, Tilman M; Simioni, Paolo; Castoldi, Elisabetta

2018-06-04

 Activated factor V (FVa) is a potent procoagulant cofactor in the prothrombinase complex, whereas its precursor factor V (FV) stimulates the inhibition of factor Xa (FXa) by tissue factor pathway inhibitor-α (TFPIα), presumably by promoting TFPIα binding to phospholipids. Plasma FV comprises two glycosylation isoforms (FV1 and FV2) with low and high phospholipid-binding affinity, respectively. The FV1/FV2 ratio is increased in carriers of the FV R2 haplotype.  This article demonstrates the TFPIα-cofactor function of FV in plasma and compares FV1 and FV2.  Thrombin generation at low TF concentration was measured in FV-depleted plasma reconstituted with 0 to 100% FV, FV1 or FV2, and in 122 individuals genotyped for the R2 haplotype. The TFPIα-cofactor activities of FV1 and FV2 were also investigated in a model system of TFPIα-mediated FXa inhibition.  In the FV titration, thrombin generation first increased (up to 5% FV) and then progressively decreased at higher FV concentrations. This anticoagulant effect of FV, which was also observed with FV2 but not with FV1, was largely abolished by anti-TFPIα antibodies, suggesting that it reflects TFPIα-cofactor activity of FV. In the model system of TFPIα-mediated FXa inhibition, FV2 was a more potent TFPIα-cofactor than FV1, in line with their respective phospholipid affinities. Accordingly, FV R2 carriers had higher thrombin generation than non-carriers, even after correction for demographics and plasma levels of coagulation factors and inhibitors.  FV (and particularly its FV2 isoform) contributes to the TFPIα-dependent down-regulation of thrombin generation in plasma triggered with low TF. Schattauer GmbH Stuttgart.

Chimeric rabbit/human Fab antibodies against the hepatitis Be-antigen and their potential applications in assays, characterization, and therapy.

PubMed

Zhuang, Xiaolei; Watts, Norman R; Palmer, Ira W; Kaufman, Joshua D; Dearborn, Altaira D; Trenbeath, Joni L; Eren, Elif; Steven, Alasdair C; Rader, Christoph; Wingfield, Paul T

2017-10-06

Hepatitis B virus (HBV) infection afflicts millions worldwide, causing cirrhosis and liver cancer. HBV e-antigen (HBeAg), a clinical marker for disease severity, is a soluble variant of the viral capsid protein. HBeAg is not required for viral replication but is implicated in establishing immune tolerance and chronic infection. The structure of recombinant e-antigen (rHBeAg) was recently determined, yet to date, the exact nature and quantitation of HBeAg still remain uncertain. Here, to further characterize HBeAg, we used phage display to produce a panel of chimeric rabbit/human monoclonal antibody fragments (both Fab and scFv) against rHBeAg. Several of the Fab/scFv, expressed in Escherichia coli , had unprecedentedly high binding affinities ( K d ∼10 -12 m) and high specificity. We used Fab/scFv in the context of an enzyme-linked immunosorbent assay (ELISA) for HBeAg quantification, which we compared with commercially available kits and verified with seroconversion panels, the WHO HBeAg standard, rHBeAg, and patient plasma samples. We found that the specificity and sensitivity are superior to those of existing commercial assays. To identify potential fine differences between rHBeAg and HBeAg, we used these Fabs in microscale immunoaffinity chromatography to purify HBeAg from individual patient plasmas. Western blotting and MS results indicated that rHBeAg and HBeAg are essentially structurally identical, although HBeAg from different patients exhibits minor carboxyl-terminal heterogeneity. We discuss several potential applications for the humanized Fab/scFv.

Foamy virus reverse transcriptase is expressed independently from the Gag protein.

PubMed Central

Enssle, J; Jordan, I; Mauer, B; Rethwilm, A

1996-01-01

In the foamy virus (FV) subgroup of retroviruses the pol genes are located in the +1 reading frame relative to the gag genes and possess potential ATG initiation codons in their 5' regions. This genome organization suggests either a + 1 ribosomal frameshift to generate a Gag-Pol fusion protein, similar to all other retroviruses studied so far, or new initiation of Pol translation, as used by pararetroviruses, to express the Pol protein. By using a genetic approach we have ruled out the former possibility and provide evidence for the latter. Two down-mutations (M53 and M54) of the pol ATG codon were found to abolish replication and Pol protein expression of the human FV isolate. The introduction of a new ATG in mutation M55, 3' to the down-mutated ATG of mutation M53, restored replication competence, indicating that the pol ATG functions as a translational initiation codon. Two nonsense mutants (M56 and M57), which functionally separated gag and pol with respect to potential frame-shifting sites, were also replication-competent, providing further genetic evidence that FVs express the Pol protein independently from Gag. Our results show that during a particular step of the replication cycle, FVs differ fundamentally from all other retroviruses. Images Fig. 3 PMID:8633029

[Isolation and identification of mosquito-borne arboviruses in Yuncheng city, Shanxi province, 2012].

PubMed

Zheng, Yayun; Cao, Yuxi; Fu, Shihong; Cheng, Jingxia; Zhao, Junying; Dai, Peifang; Kong, Xiangsheng; Liang, Guodong

2015-04-01

To investigate the species and distribution of mosquitoes and mosquito-borne arboviruses in Yuncheng city of Shanxi province, China. Mosquito samples were collected in 19 collection sites from Linyi county and Yongji city in Yuncheng city, in August, 2012. After identification and classification, all the specimens were homogenized and centrifuged to acquire supernatant before being inoculated to both C6/36 and BHK21 cells for viral isolation. Positive isolates were identified with arbovirus species-specific primers under RT-PCR, for further sequencing and phylogenetic analysis. A total of 10 455 mosquitoes of 7 species in 4 genuese were collected. The predominant mosquito species in Linyi county was Culex pipens pallens (91.96%, 3 911/4 253), but the one in Yongji city was Culex tritaeniorhynchus (72.85%, 4 518/6 202). A total of 23 strains of viruses were isolated from the mosquito pools. 15 strains from Culex tritaeniorhynchus and Culex pipens pallens were identified as genotype I Japanese encephalitis virus (JEV). Four strains from Culex pipens pallens were identified as Culex flavivirus (CxFV). Three strains from Culex pipens pallens were identified as Culex pipiens pallens densovirus (CppDNV). One strain from Armigeres subalbatus and Aedes albopictus was identified as Getah virus (GETV). Four kinds of arboviruses were isolated from the mosquito pools, including GETV and CxFV, which were isolated and documented in Shanxi province for the first time. In the city of Yuncheng, Culex tritaeniorhynchus had been the predominant species and major vector for transmitting JEV. Genotype I JEV remained the major JEV circulating in the local natural environment.

Induction of complex immune responses and strong protection against retrovirus challenge by adenovirus-based immunization depends on the order of vaccine delivery.

PubMed

Kaulfuß, Meike; Wensing, Ina; Windmann, Sonja; Hrycak, Camilla Patrizia; Bayer, Wibke

2017-02-06

In the Friend retrovirus mouse model we developed potent adenovirus-based vaccines that were designed to induce either strong Friend virus GagL 85-93 -specific CD8 + T cell or antibody responses, respectively. To optimize the immunization outcome we evaluated vaccination strategies using combinations of these vaccines. While the vaccines on their own confer strong protection from a subsequent Friend virus challenge, the simple combination of the vaccines for the establishment of an optimized immunization protocol did not result in a further improvement of vaccine effectivity. We demonstrate that the co-immunization with GagL 85-93 /leader-gag encoding vectors together with envelope-encoding vectors abrogates the induction of GagL 85-93 -specific CD8 + T cells, and in successive immunization protocols the immunization with the GagL 85-93 /leader-gag encoding vector had to precede the immunization with an envelope encoding vector for the efficient induction of GagL 85-93 -specific CD8 + T cells. Importantly, the antibody response to envelope was in fact enhanced when the mice were adenovirus-experienced from a prior immunization, highlighting the expedience of this approach. To circumvent the immunosuppressive effect of envelope on immune responses to simultaneously or subsequently administered immunogens, we developed a two immunizations-based vaccination protocol that induces strong immune responses and confers robust protection of highly Friend virus-susceptible mice from a lethal Friend virus challenge.

Detection of Culex flavivirus and Aedes flavivirus nucleotide sequences in mosquitoes from parks in the city of São Paulo, Brazil.

PubMed

Fernandes, Licia Natal; de Paula, Marcia Bicudo; Araújo, Alessandra Bergamo; Gonçalves, Elisabeth Fernandes Bertoletti; Romano, Camila Malta; Natal, Delsio; Malafronte, Rosely dos Santos; Marrelli, Mauro Toledo; Levi, José Eduardo

2016-05-01

The dengue viruses are widespread in Brazil and are a major public health concern. Other flaviviruses also cause diseases in humans, although on a smaller scale. The city of São Paulo is in a highly urbanized area with few green spaces apart from its parks, which are used for recreation and where potential vertebrate hosts and mosquito vectors of pathogenic Flavivirus species can be found. Although this scenario can contribute to the transmission of Flavivirus to humans, little is known about the circulation of members of this genus in these areas. In light of this, the present study sought to identify Flavivirus infection in mosquitoes (Diptera: Culicidae) collected in parks in the city of São Paulo. Seven parks in different sectors of the city were selected. Monthly mosquito collections were carried out in each park from March 2011 to February 2012 using aspiration and traps (Shannon and CD C-CO2). Nucleic acids were extracted from the mosquitoes collected and used for reverse-transcriptase and real-time polymerase chain reactions with genus-specific primers targeting a 200-nucleotide region in the Flavivirus NS5 gene. Positive samples were sequenced, and phylogenetic analyses were performed. Culex and Aedes were the most frequent genera of Culicidae collected. Culex flavivirus (CxFV)-related and Aedes flavivirus (AEFV)- related nucleotide sequences were detected in 17 pools of Culex and two pools of Aedes mosquitoes, respectively, among the 818 pools of non-engorged females analyzed. To the best of our knowledge, this is the first report of CxFV and AEFV in the city of São Paulo and Latin America, respectively. Both viruses are insect- specific flaviviruses, a group known to replicate only in mosquito cells and induce a cytopathic effect in some situations. Hence, our data suggests that CxFV and AEFV are present in Culex and Aedes mosquitoes, respectively, in parks in the city of São Paulo. Even though Flavivirus species of medical importance were not detected, surveillance is recommended in the study areas because of the presence of vertebrates and mosquitoes that could act as amplifying hosts and vectors of flaviviruses, providing the required conditions for circulation of these viruses. Copyright © 2016 Elsevier B.V. All rights reserved.

Supercritical anti-solvent technique assisted synthesis of thymoquinone liposomes for radioprotection: Formulation optimization, in-vitro and in-vivo studies.

PubMed

Ahmad, Iqbal; Akhter, Sohail; Anwar, Mohammed; Zafar, Sobiya; Sharma, Rakesh Kumar; Ali, Asgar; Ahmad, Farhan Jalees

2017-05-15

The aim of this study was to develop Thymoquinone (TQ) loaded PEGylated liposomes using supercritical anti-solvent (SAS) process for enhanced blood circulation, and greater radioprotection. The SAS process of PEGylated liposomes synthesis was optimized by Box-Behnken design. Spherical liposomes with a particle size of 195.6±5.56nm and entrapment efficiency (%EE) of 89.4±3.69% were obtained. Optimized SAS process parameters; temperature, pressure and solution flow rate were 35°C, 140bar and 0.18mL/min, respectively, while 7.5mmol phospholipid, 0.75mmol of cholesterol, and 1mmol TQ were optimized formulation ingredients. Incorporation of MPEG-2000-DSPE (5% w/w) provided the PEGylated liposomes (FV-17B; particle size=231.3±6.74nm, %EE=91.9±3.45%, maximum TQ release >70% in 24h). Pharmacokinetics of FV-17B in mice demonstrated distinctly superior systemic circulation time for TQ in plasma. Effectiveness of radioprotection by FV-17B in mice model was demonstrated by non-significant body weight change, normal vital blood components (WBCs, RBCs, and Platelets), micronuclei and spleen index and increased survival probability in post irradiation animal group as compared to controls (plain TQ and marketed formulation). Altogether, the results anticipated that the SAS process could serve as a single step environmental friendly technique for the development of stable long circulating TQ loaded liposomes for effective radioprotection. Copyright © 2017 Elsevier B.V. All rights reserved.

Endocytosis of exogenous factor V by ex-vivo differentiated megakaryocytes from patients with severe parahaemophilia.

PubMed

Radu, Claudia M; Spiezia, Luca; Bulato, Cristiana; Gavasso, Sabrina; Campello, Elena; Sartorello, Francesca; Castoldi, Elisabetta; Simioni, Paolo

2016-11-01

Although human megakaryocytes can synthesize factor V (FV), platelet FV derives largely from endocytosis of plasma FV. Recently, it has been shown that plasma transfusions can replenish the platelet FV pool in parahaemophilic patients. Here we corroborate this finding by showing FV endocytosis by ex vivo differentiated megakaryocytes derived from patients with inherited parahaemophilia. Mononuclear stem cells isolated from peripheral blood of healthy subjects and of three patients with severe parahaemophilia were cultured in the presence of thrombopoietin and interleukin-3 and differentiated into CD41-positive polynucleated megakaryocytes. Exogenous purified FV was added to the culture medium to evaluate FV endocytosis. Immunofluorescence staining revealed abundant FV expression in megakaryocytes derived from healthy donors, but no FV expression in those derived from patients with severe parahaemophilia. However, after the addition of purified FV to the culture medium, megakaryocytes from parahaemophilia patients became positive upon FV immunostaining, suggesting endocytosis of exogenous FV. Endocytosed FV retained factor Xa-co-factor activity as assessed by a prothrombin time-based functional test in megakaryocyte lysates. Addition of exogenous FV to culture medium can restore the FV content of megakaryocytes derived from patients with severe FV defects. This rescue mechanism can have important clinical implications in the management of parahaemophilia patients. © 2016 John Wiley & Sons Ltd.

Insertion of targeting domains into the envelope glycoprotein of Moloney murine leukemia virus (MoMLV)-based vectors modulates the route of mCAT-1-mediated viral entry.

PubMed

Viejo-Borbolla, A; Pizzato, M; Blair, E D; Schulz, T F

2005-03-01

Several groups have inserted targeting domains into the envelope glycoprotein (Env) of Moloney murine leukemia virus (MoMLV) in an attempt to produce targeted retroviral vectors for human gene therapy. While binding of these modified Envs to the target molecule expressed on the surface of human cells was observed, specific high-titer infection of human cells expressing the target molecule was not achieved. Here we investigate the initial steps in the entry process of targeted MoMLV vectors both in murine and human cells expressing the MoMLV receptor, the mouse cationic amino acid transporter-1 (mCAT-1). We show that insertion of a small ligand targeted to E-selectin and of a single chain antibody (scFv) targeted to folate-binding protein (FBP) into the N-terminus of MoMLV Env results in the reduction of the infectivity and the kinetics of entry of the MoMLV vectors. The use of soluble receptor-binding domain (sRBD), bafilomycin A1 (BafA1) and methyl-beta-cyclodextrin (MbetaC) increase the infectivity of the MoMLV vectors targeted to FBP (MoMLV-FBP) suggesting that the scFv targeted to FBP increases the threshold for fusion and might re-route entry of the targeted MoMLV-FBP vector towards an endocytic, non-productive pathway.

Analysis of Proteins of Mouse Sarcoma Pseudotype Viruses: Type-Specific Radioimmunoassays for Ecotropic Virus p30's

PubMed Central

Kennel, Stephen J.; Tennant, Raymond W.

1979-01-01

Murine sarcoma virus pseudotypes were prepared by infection of nonproducer cells (A1-2), which were transformed by the Gazdar strain of mouse sarcoma virus, with Gross (N-tropic), WN1802B (B-tropic), or Moloney (NB-tropic) viruses. The respective host range pseudotype sarcoma viruses were defined by the titration characteristics on cells with the appropriate Fv-1 genotype. Proteins from virus progeny were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Bands present in both the 65,000- and the 10,000- to 20,000- molecular-weight regions of the gel distinguished the pseudotype viruses from their respective helpers. Furthermore, two protein bands were noted in the p30 region of murine sarcoma virus (Gross), one corresponding to Gross virus p30, and another of slightly slower mobility. However, since the mobility of the putative sarcoma p30 is nearly indentical to that of WN1802B, its presence could not be established by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Type-specific radioimmunoassays for Gross virus p30 and for WN1802B p30 were applied for analysis of pseudotype preparations, and among several ecotropic viruses tested, only the homologous virus scored in the respective assay. By use of these assays, pseudotype viruses were found to contain only 8 to 48% helper-specific p30's; the remainder is presumably derived from the sarcoma virus. Images PMID:90164

Implementing a free school-based fruit and vegetable programme: barriers and facilitators experienced by pupils, teachers and produce suppliers in the Boost study

PubMed Central

2014-01-01

Background Multi-component interventions which combine educational and environmental strategies appear to be most effective in increasing fruit and vegetable (FV) intake in adolescents. However, multi-component interventions are complex to implement and often poorly implemented. Identification of barriers and facilitators for implementation is warranted to improve future interventions. This study aimed to explore implementation of two intervention components which addressed availability and accessibility of FV in the multi-component, school-based Boost study which targeted FV intake among Danish 13-year-olds and to identify barriers and facilitators for implementation among pupils, teachers and FV suppliers. Methods We conducted focus group interviews with 111 13-year-olds and 13 teachers, completed class observations at six schools, and conducted telephone interviews with all involved FV suppliers. Interviews were transcribed, coded and analysed using qualitative analytical procedures. Results FV suppliers affected the implementation of the FV programme at schools and thereby pupils’ intake through their timing of delivery and through the quality, quantity and variety of the delivered FV. Teachers influenced the accessibility and appearance of FV by deciding if and when the pupils could eat FV and whether FV were cut up. Different aspects of time acted as barriers for teachers’ implementation of the FV programme: time spent on having a FV break during lessons, time needed to prepare FV and time spent on pupils’ misbehaviour and not being able to handle getting FV. Teacher timing of cutting up and serving FV could turn into a barrier for pupils FV intake due to enzymatic browning. The appearance of FV was important for pupils’ intake, especially for girls. FV that did not appeal to the pupils e.g. had turned brown after being cut up were thrown around as a part of a game by the pupils, especially boys. Girls appreciated the social dimension of eating FV together to a larger extent than boys. Conclusions Limited time and pupils’ misbehaviour were barriers for teachers’ implementation. Establishing FV delivery to schools as a new routine challenged FV suppliers’ implementation. Food aesthetics were important for most pupils’ FV intake while the social dimension of eating FV together seemed more important to girls than boys. Trial registration Current Controlled Trials ISRCTN11666034. PMID:24512278

Implementing a free school-based fruit and vegetable programme: barriers and facilitators experienced by pupils, teachers and produce suppliers in the Boost study.

PubMed

Aarestrup, Anne Kristine; Krølner, Rikke; Jørgensen, Thea Suldrup; Evans, Alexandra; Due, Pernille; Tjørnhøj-Thomsen, Tine

2014-02-11

Multi-component interventions which combine educational and environmental strategies appear to be most effective in increasing fruit and vegetable (FV) intake in adolescents. However, multi-component interventions are complex to implement and often poorly implemented. Identification of barriers and facilitators for implementation is warranted to improve future interventions.This study aimed to explore implementation of two intervention components which addressed availability and accessibility of FV in the multi-component, school-based Boost study which targeted FV intake among Danish 13-year-olds and to identify barriers and facilitators for implementation among pupils, teachers and FV suppliers. We conducted focus group interviews with 111 13-year-olds and 13 teachers, completed class observations at six schools, and conducted telephone interviews with all involved FV suppliers. Interviews were transcribed, coded and analysed using qualitative analytical procedures. FV suppliers affected the implementation of the FV programme at schools and thereby pupils' intake through their timing of delivery and through the quality, quantity and variety of the delivered FV. Teachers influenced the accessibility and appearance of FV by deciding if and when the pupils could eat FV and whether FV were cut up. Different aspects of time acted as barriers for teachers' implementation of the FV programme: time spent on having a FV break during lessons, time needed to prepare FV and time spent on pupils' misbehaviour and not being able to handle getting FV. Teacher timing of cutting up and serving FV could turn into a barrier for pupils FV intake due to enzymatic browning. The appearance of FV was important for pupils' intake, especially for girls. FV that did not appeal to the pupils e.g. had turned brown after being cut up were thrown around as a part of a game by the pupils, especially boys. Girls appreciated the social dimension of eating FV together to a larger extent than boys. Limited time and pupils' misbehaviour were barriers for teachers' implementation. Establishing FV delivery to schools as a new routine challenged FV suppliers' implementation. Food aesthetics were important for most pupils' FV intake while the social dimension of eating FV together seemed more important to girls than boys. Current Controlled Trials ISRCTN11666034.

Infection in systemic lupus erythematosus: friend or foe?

PubMed Central

Francis, Lisa; Perl, Andras

2010-01-01

Infectious agents have long been implicated in the pathogenesis of systemic lupus erythematosus. Common viruses, such as the Epstein-Barr virus, transfusion transmitted virus, parvovirus and cytomegalovirus, have an increased prevalence in patients with systemic lupus erythematosus. They may contribute to disease pathogenesis through triggering autoimmunity via structural or functional molecular mimicry, encoding proteins that induce cross-reactive immune responses to self antigens or modulate antigen processing, activation, or apoptosis of B and T cells, macrophages or dendritic cells. Alternatively, some infectious agents, such as malaria, Toxoplasma gondii and Helicobacter pylori, may have a protective effect. Vaccinations may play dual roles by protecting against friend and foe alike. PMID:20209114

Activity of Pure Streptovaricins and Fractionated Streptovaricin Complex Against Friend Virus

PubMed Central

Horoszewicz, Julius S.; Rinehart, Kenneth L.; Leong, Susan S.; Carter, William A.

1975-01-01

Chromatographic fractionation of streptovaricin complex yields two stable components enriched (4- to 16-fold) in activity directed against the polycythemic strain of Friend virus; both components apparently contain no streptovaricins. When compared with their unfractionated parent streptovaricin complex, eight individual intact streptovaricins (A through G and J) show at least a 30-fold reduction in antiviral activity. These results further support the conclusion that the diversified biological properties of streptovaricin complex probably reside in different molecular structures. PMID:237470

High Specific Selectivity and Membrane-Active Mechanism of Synthetic Cationic Hybrid Antimicrobial Peptides Based on the Peptide FV7

PubMed Central

Tan, Tingting; Wu, Di; Li, Weizhong; Zheng, Xin; Li, Weifen; Shan, Anshan

2017-01-01

Hybrid peptides integrating different functional domains of peptides have many advantages, such as remarkable antimicrobial activity, lower hemolysis and ideal cell selectivity, compared with natural antimicrobial peptides. FV7 (FRIRVRV-NH2), a consensus amphiphilic sequence was identified as being analogous to host defense peptides. In this study, we designed a series of hybrid peptides FV7-LL-37 (17–29) (FV-LL), FV7-magainin 2 (9–21) (FV-MA) and FV7-cecropin A (1–8) (FV-CE) by combining the FV7 sequence with the small functional sequences LL-37 (17–29) (LL), magainin 2 (9–21) (MA) and cecropin A (1–8) (CE) which all come from well-described natural peptides. The results demonstrated that the synthetic hybrid peptides, in particular FV-LL, had potent antibacterial activities over a wide range of Gram-negative and Gram-positive bacteria with lower hemolytic activity than other peptides. Furthermore, fluorescent spectroscopy indicated that the hybrid peptide FV-LL exhibited marked membrane destruction by inducing outer and inner bacterial membrane permeabilization, while scanning electron microscopy (SEM) and transmission electron microscopy (TEM) demonstrated that FV-LL damaged membrane integrity by disrupting the bacterial membrane. Inhibiting biofilm formation assays also showed that FV-LL had similar anti-biofilm activity compared with the functional peptide sequence FV7. Synthetic cationic hybrid peptides based on FV7 could provide new models for combining different functional domains and demonstrate effective avenues to screen for novel antimicrobial agents. PMID:28178190

Genome-Wide Identification and Characterization of Novel Laccase Genes in the White-Rot Fungus Flammulina velutipes

PubMed Central

Kim, Hong-Il; Kwon, O-Chul; Kong, Won-Sik; Lee, Chang-Soo

2014-01-01

The aim of this study was to identify and characterize new Flammulina velutipes laccases from its whole-genome sequence. Of the 15 putative laccase genes detected in the F. velutipes genome, four new laccase genes (fvLac-1, fvLac-2, fvLac3, and fvLac-4) were found to contain four complete copper-binding regions (ten histidine residues and one cysteine residue) and four cysteine residues involved in forming disulfide bridges, fvLac-1, fvLac-2, fvLac3, and fvLac-4, encoding proteins consisting of 516, 518, 515, and 533 amino acid residues, respectively. Potential N-glycosylation sites (Asn-Xaa-Ser/Thr) were identified in the cDNA sequence of fvLac-1 (Asn-454), fvLac-2 (Asn-437 and Asn-455), fvLac-3 (Asn-111 and Asn-237), and fvLac4 (Asn-402 and Asn-457). In addition, the first 19~20 amino acid residues of these proteins were predicted to comprise signal peptides. Laccase activity assays and reverse transcription polymerase chain reaction analyses clearly reveal that CuSO4 affects the induction and the transcription level of these laccase genes. PMID:25606003

Associations between school meals offered through the National School Lunch Program and the School Breakfast Program and fruit and vegetable intake among ethnically diverse, low-income children.

PubMed

Robinson-O'Brien, Ramona; Burgess-Champoux, Teri; Haines, Jess; Hannan, Peter J; Neumark-Sztainer, Dianne

2010-10-01

Despite evidence in support of the health benefits associated with fruit and vegetable (FV) intake, national data indicate that FV consumption among school-aged children is below recommended levels, particularly among low-income children. School meals offered through the School Breakfast Program and National School Lunch Program can provide an important contribution to child FV intake. This study examines the proportion of fruits and vegetables consumed from school meals programs among ethnically diverse, low socioeconomic status children. Participants (n = 103) included fourth to sixth grade boys and girls from 4 urban elementary schools in St. Paul, Minnesota serving primarily low-income populations. Research staff interviewed children during school hours and recorded dietary intake via 24-hour recall. Analysis included descriptive statistics using cross tabulations and means. Average reported mean (SD) daily FV intake was 3.6 (2.5) servings, with 80% of children consuming fewer than 5 daily servings of FV. On average, children consumed over half of their daily FV intake within school. Children with low FV intake (<5 FV servings daily) consumed a higher proportion of their daily intake at school than children with higher FV intake (≥5 FV servings daily) (39% vs 59%; p = .002). Child FV intake is below recommended levels. School meals provide an important contribution to the daily FV intake among ethnically diverse, low socioeconomic status children, particularly among those with the lowest FV intake. School meals programs promoting FV intake within the school environment may provide an opportunity to encourage increased FV consumption. © 2010, American School Health Association.

Fruit and Vegetable Attitudes, Norms, and Intake in Low-Income Youth.

PubMed

Di Noia, Jennifer; Cullen, Karen Weber

2015-12-01

Fruit and vegetable (FV) attitudes and norms have been shown to influence intake in youth; yet research with low-income youth and studies supplementing self-report with objective measures of intake are lacking. Cross-sectional survey data on self-rated FV intake, FV attitudes, and FV norms were collected in a sample of 116 youth attending a residential summer camp serving low-income families. FV intake also was estimated by direct observation. Differences between self-rated and observed FV intake, perceived and observed peer intake, and perceived and peer-reported attitudes toward eating FVs were assessed with paired samples t tests. The role of FV attitudes, descriptive norms (perceived peer FV intake), injunctive norms (perceived peer attitudes toward eating FVs), and actual norms (observed peer FV intake and peer-reported FV attitudes) in predicting FV intake also was examined with multiple regression analysis. Youth misperceived their own and their peers' FV intake (i.e., overestimated intake of fruit and underestimated intake of vegetables) and believed that peers held less favorable attitudes toward eating FVs than was the case. The models predicting self-rated intake were significant, accounting for 34% of the variance in fruit intake and 28% of the variance in vegetable intake. Attitudes and descriptive norms were positively associated with FV intake, and observed peer fruit intake was negatively associated with fruit intake. Findings suggest that in low-income youth, FV attitudes, descriptive norms, and normative peer behavior predict perceived but not actual intake. Youth may benefit from intervention to promote favorable FV attitudes and norms. A focus on descriptive norms holds promise for improving self-rated intake in this population. © 2015 Society for Public Health Education.

The effectiveness of asking behaviors among 9-11 year-old children in increasing home availability and children's intake of fruit and vegetables: results from the Squire's Quest II self-regulation game intervention.

PubMed

DeSmet, Ann; Liu, Yan; De Bourdeaudhuij, Ilse; Baranowski, Tom; Thompson, Debbe

2017-04-21

Home environment has an important influence on children's fruit and vegetable (FV) consumption, but children may in turn also impact their home FV environment, e.g. by asking for FV. The Squire's Quest II serious game intervention aimed to increase asking behaviors to improve home FV availability and children's FV intake. This study's aims were to assess: 1) did asking behaviors at baseline predict home FV availability at baseline (T0) (RQ1); 2) were asking behaviors and home FV availability influenced by the intervention (RQ2); 3) did increases in asking behaviors predict increased home FV availability (RQ3); and 4) did increases in asking behaviors and increases in home FV availability mediate increases in FV intake among children (RQ4)? This is a secondary analysis of a study using a randomized controlled trial, with 4 groups (each n = 100 child-parent dyads). All groups were analyzed together for this paper since groups did not vary on components relevant to our analysis. All children and parents (n = 400 dyads) received a self-regulation serious game intervention and parent material. The intervention ran for three months. Measurements were taken at baseline, immediately after intervention and at 3-month follow-up. Asking behavior and home FV availability were measured using questionnaires; child FV intake was measured using 24-h dietary recalls. ANCOVA methods (research question 1), linear mixed-effect models (research question 2), and Structural Equation Modeling (research questions 3 and 4) were used. Baseline child asking behaviors predicted baseline home FV availability. The intervention increased child asking behaviors and home FV availability. Increases in child asking behaviors, however, did not predict increased home FV availability. Increased child asking behaviors and home FV availability also did not mediate the increases in child FV intake. Children influence their home FV environment through their asking behaviors, which can be enhanced via a serious game intervention. The obtained increases in asking behavior were, however, insufficient to affect home FV availability or intake. Other factors, such as child preferences, sample characteristics, intervention duration and parental direct involvement may play a role and warrant examination in future research. ClinicalTrials.gov NCT01004094 . Date registered 10/28/2009.

Selecting and expressing protective single-chain Fv fragment to stabilize L-asparaginase against inactivation by trypsin.

PubMed

Guo, L; Yan, X; Qian, S; Meng, G

2000-02-01

Four non-inhibitory specific single-chain Fv (sc Fv) fragments directed against L-asparaginase (ASNase) of Escherichia coli were selected from a synthetic phage-display scFv library. The scFv46 fragment could enhance the resistance of ASNase to trypsin proteolysis, with 70% of the initial ASNase activity present after the ASNase-scFv46 complex had been treated with trypsin for 30 min at 37 degrees C, whereas little residual activity was detected without the scFv46 fragment. The scFv46 gene was cloned to an expression vector pET-21a and expressed at high levels (about 45% of total cell protein) in E. coli BL21 (DE3) as inclusion bodies. The refolded and purified scFv46 fragment was proved to protect ASNase, and the protective effect was further confirmed by SDS/PAGE. It was found that under optimum conditions of molar ratio of scFv to ASNase, incubation time and temperature, the residual activity of the ASNase-scFv46 complex could reach about 78% after treatment with trypsin for 30 min at 37 degrees C. The results demonstrated that scFv fragments prepared by phage-antibody library technology could be used to protect target proteins.

Fruits and vegetables displace, but do not decrease, total energy in school lunches.

PubMed

Bontrager Yoder, Andrea B; Schoeller, Dale A

2014-08-01

The high overweight and obesity prevalence among US children is a well-established public health concern. Diet is known to play a causal role in obesity. Increasing fruit and vegetable (FV) consumption to recommended levels is proposed to help reduce obesity, because their bulk and low energy density are believed to reduce energy-dense food consumption (volume displacement hypothesis). This study tests this hypothesis at the lunch meal among upper-elementary students participating in a Farm to School (F2S) program. Digital photographs of students' school lunch trays were visually analyzed to identify the food items and amounts that were present and consumed before and after the meal. Using the USDA Nutrient Database, total and FV-only energy were calculated for each tray. Analysis of total- and non-FV energy intake was performed according to (1) levels of FV energy intake, (2) FV energy density, and (3) previous years of Farm to School programming. Higher intake of FV energy displaced non-FV energy, but total energy did not decrease across FV energy intake groups. High-FV-energy-density trays showed lower non-FV energy intake than low-FV-energy-density trays (470±179 vs. 534±219 kcal; p<0.0001). Trays from schools with more previous years of F2S programming decreased total and non-FV energy intake from school lunches (p for trend<0.0001, both). Increased FV consumption reduces non-FV energy intake, but does not reduce total energy intake. Therefore, this study does not support the volume displacement hypothesis and suggests calorie displacement instead.

Family Violence and Associated Help-Seeking Behavior among Older African American Women

PubMed Central

Paranjape, Anuradha; Tucker, Alyce; Mckenzie-Mack, LaTasha; Thompson, Nancy; Kaslow, Nadine

2007-01-01

Objective Little is known about how older African American women define family violence (FV) and what FV survivors might expect from their healthcare providers. The purpose of this study was to understand how these women define FV, where they seek help for FV, and what barriers they face in these efforts. Methods We conducted 6 focus groups with 30 African American women over the age of 50, including some FV survivors, at a large, inner-city public hospital. Results Participants defined FV broadly, citing examples of abuse (physical, sexual, emotional and financial) and neglect. Spiritual sources were cited over physicians as being available to help FV survivors. Barriers to receiving assistance included negative encounters with physicians, lack of trust in the system and dearth of age-appropriate resources. Conclusions For older African American women, FV takes many forms of which many may not be obvious during the clinical encounter. Like younger FV survivors, they expect physicians to serve as a resource for FV. Practice implications Physicians caring for older African American women need to remember to ask them about FV, and when making referrals for abuse and neglect, consider offering referrals to pastoral care if appropriate. PMID:17644300

A conjugate of an anti-midkine single-chain variable fragment to doxorubicin inhibits tumor growth

PubMed Central

Zhao, Shuli; Zhao, Guangfeng; Xie, Hao; Huang, Yahong; Hou, Yayi

2012-01-01

Doxorubicin (DOX) was conjugated to a single-chain variable fragment (scFv) against human midkine (MK), and the conjugate (scFv-DOX) was used to target the chemotherapeutic agent to a mouse solid tumor model in which the tumor cells expressed high levels of human MK. The His-tagged recombinant scFv was expressed in bacteria, purified by metal affinity chromatography, and then conjugated to DOX using oxidative dextran (Dex) as a linker. The molecular formula of this immunoconjugate was scFv(Dex)1.3(DOX)20. In vitro apoptosis assays showed that the scFv-DOX conjugate was more cytotoxic against MK-transfected human adenocarcinoma cells (BGC823-MK) than untransfected cells (55.3 ± 2.4 vs 22.4 ± 3.8%) for three independent experiments. Nude mice bearing BGC823-MK solid tumors received scFv-DOX or equivalent doses of scFv + DOX for 2 weeks and tumor growth was more effectively inhibited by the scFv-DOX conjugate than by scFv + DOX (51.83% inhibition vs 40.81%). Histological analysis of the tumor tissues revealed that the highest levels of DOX accumulated in tumors from mice treated with scFv-DOX and this resulted in more extensive tumor cell death than in animals treated with the equivalent dose of scFv + DOX. These results show that the scFv-DOX conjugate effectively inhibited tumor growth in vivo and suggest that antigen-specific scFv may be competent drug-carriers. PMID:22267001

Family meals can help children reach their 5 a day: a cross-sectional survey of children's dietary intake from London primary schools.

PubMed

Christian, Meaghan S; Evans, Charlotte E L; Hancock, Neil; Nykjaer, Camilla; Cade, Janet E

2013-04-01

This study aims to explore how the home food environment and parental attitudes and values affect children's fruit and vegetable (F&V) intake. The sample consists of 2383 children with a mean age of 8.3 years (95% CI 8.2 to 8.3) attending 52 primary schools in London. These children are taking part in two randomised controlled trials to evaluate a school gardening programme. Diet was assessed using a validated 24-h food tick list, the Child And Diet Evaluation Tool (CADET). The CADET tool found that children consumed on average 293 g F&V (95% CI 287 to 303) per day. Clustered (by school) multilevel regression models with total F&V as the primary outcome were conducted to explore how the home environment affects children's F&V intake. Children of families who reported 'always' eating a family meal together at a table had 125 g (95% CI 92 to 157; p=<0.001) more F&V than families who never ate a meal together. Daily consumption of F&V by parents was associated with higher F&V (88 g, 95% CI 37 to 138) intake in children compared with rarely/never consumption of F&V by parents. Cutting up fruit and vegetables for children was associated with higher consumption. Families who reported always cutting up F&V for their children had 44 g (95% CI 18 to 71) more F&V than families who never cut up F&V. This study identified that cutting up F&V and family consumption of F&V facilitates children's intake. Eating a family meal together regularly could increase children's F&V intake and help them achieve the recommended intake. ISRCTN11396528.

Fruits and Vegetables Displace, But Do Not Decrease, Total Energy in School Lunches

PubMed Central

Schoeller, Dale A.

2014-01-01

Abstract Background: The high overweight and obesity prevalence among US children is a well-established public health concern. Diet is known to play a causal role in obesity. Increasing fruit and vegetable (FV) consumption to recommended levels is proposed to help reduce obesity, because their bulk and low energy density are believed to reduce energy-dense food consumption (volume displacement hypothesis). This study tests this hypothesis at the lunch meal among upper-elementary students participating in a Farm to School (F2S) program. Methods: Digital photographs of students' school lunch trays were visually analyzed to identify the food items and amounts that were present and consumed before and after the meal. Using the USDA Nutrient Database, total and FV-only energy were calculated for each tray. Analysis of total- and non-FV energy intake was performed according to (1) levels of FV energy intake, (2) FV energy density, and (3) previous years of Farm to School programming. Results: Higher intake of FV energy displaced non-FV energy, but total energy did not decrease across FV energy intake groups. High-FV-energy-density trays showed lower non-FV energy intake than low-FV-energy-density trays (470±179 vs. 534±219 kcal; p<0.0001). Trays from schools with more previous years of F2S programming decreased total and non-FV energy intake from school lunches (p for trend<0.0001, both). Conclusions: Increased FV consumption reduces non-FV energy intake, but does not reduce total energy intake. Therefore, this study does not support the volume displacement hypothesis and suggests calorie displacement instead. PMID:24988122

Technology of compact MAb and its application for medicinal plant breeding named as missile type molecular breeding.

PubMed

Putalun, Waraporn

2011-03-01

Single chain fragment-variable (scFv) enhanced solasodine glycoside accumulation in Solanum khasianum hairy root cultures transformed by the ScFv solamargine (As)-scFv gene. The scFv protein was expressed at a high level in inclusion bodies of E. coli. After being renatured, the scFv protein was purified in a one-step manner by metal chelate affinity chromatography. The yield of refolded and purified scFv was 12.5 mg per 100 ml of cell culture. The characteristics of the As-scFv expressed in E. coli and transgenic hairy roots were similar to those of the parent monoclonal antibody (MAb). The expression of scFv protein provides a low cost and a high yield of functional scFv antibody against solamargine. The full linear range of the ELISA assay using scFv was extended from 1.5-10 µg/ml. The expressed anti-solamargine scFv protein could be useful for determination of total solasodine glycoside content in plant samples by ELISA. Solasodine glycoside levels in the transgenic hairy root were 2.3-fold higher than that in the wild-type hairy root based on the soluble protein level and binding activities. The As-scFv expressed in S. khasianum hairy roots enhanced solasodine glycosides accumulation and provide a novel medicinal plant breeding methodology that can produce a high yield of secondary metabolites.

What determines the fruit and vegetables intake of primary school children? - An analysis of personal and social determinants.

PubMed

Haß, Julia; Hartmann, Monika

2018-01-01

The high prevalence of childhood obesity is a major concern in developed and developing countries. An increase in fruit and vegetable (F&V) intake is perceived as one of the numerous strategies to prevent and reduce the risk of adiposity. The purpose of this study was to investigate the relevance of personal and social determinants in explaining children's F&V intake. Written questionnaire data were collected from 702 parent-child pairs that included 3rd and 4th graders (aged 7 to 10) and their parents. Children's F&V intake was recorded over three food records. Hierarchical linear regression models were applied to assess the impact of personal and social determinants on children's F&V intake. Regression models focusing on personal and social determinants revealed that the most promising personal determinants pertained to the knowledge of different types of F&V and preferences for F&V. Moreover, an exclusive focus on social determinants indicated that parental modeling and peer influence had significant and positive relationships with children's F&V intake, whereas verbal directives to eat F&V exhibited a significant and negative relationship. In combination, the following four personal and social determinants were demonstrated to be significant: knowledge of different types of F&V, preferences for F&V and parental modeling, all of which had positive relationships, and verbal directives to eat F&V, which had a negative impact. The results identify important associative determinants of children's F&V intake. These are in part personal and in part social and are shown by our analysis to be of equal and perhaps mutual importance. Therefore, we suggest that interventions aimed at improving children's F&V intake should address children's preferences for F&V, impart knowledge concerning the variety of F&V and encourage parents to act as role models. Copyright © 2017 Elsevier Ltd. All rights reserved.

IFP35 Is Involved in the Antiviral Function of Interferon by Association with the Viral Tas Transactivator of Bovine Foamy Virus▿

PubMed Central

Tan, Juan; Qiao, Wentao; Wang, Jian; Xu, Fengwen; Li, Yue; Zhou, Jun; Chen, Qimin; Geng, Yunqi

2008-01-01

Interferon-induced proteins (IFPs) exert multiple functions corresponding to diverse interferon signals. However, the intracellular functions of many IFPs are not fully characterized. Here, we report that IFP35, a member of the IFP family with a molecular mass of 35 kDa, can interact with the bovine Tas (BTas) regulatory protein of bovine foamy virus (BFV). The interaction involves NID2 (IFP35/Nmi homology domain) of IFP35 and the central domain of BTas. The overexpression of IFP35 disturbs the ability of BTas to activate viral-gene transcription and inhibits viral replication. The depletion of endogenous IFP35 by interfering RNA can promote the activation of BFV, suggesting an inhibitory function of IFP35 in viral-gene expression. In addition, IFP35 can interact with the homologous regulatory protein of prototype FV and arrest viral replication and repress viral transcription. Our study suggests that IFP35 may represent a novel pathway of interferon-mediated antiviral activity in host organisms that plays a role in the maintenance of FV latency. PMID:18305040

Trying versus liking fruits and vegetables: correspondence between mothers and preschoolers.

PubMed

Worobey, Harriet; Ostapkovich, Kathleen; Yudin, Kristin; Worobey, John

2010-01-01

Extensive research indicates that a diet rich in fruits and vegetables (F&V) protects against numerous illnesses in adulthood, but that most individuals, including children, consume far fewer F&V per day than is recommended. Since evidence suggests that eating habits in childhood track into adulthood, more research is necessary to learn about how parental F&V intake and opportunities influence child F&V consumption. The purpose of this study was to examine the relationship between mothers' F&V preferences and those of their preschool-age children to determine if greater maternal "liking" of fruits and vegetables was associated with their reports of their children's "trying" more fruits and vegetables. Eighty-three mothers completed a questionnaire that assessed whether they and their preschoolers had tried or liked a variety of F&V. Mothers liked 86% of the fruits they tried, girls 76%, and boys 69%. Mothers liked 81% of the vegetables they tried; boys and girls liked 55%. Mothers' likes correlated with F&V that their children tried, but mothers' likes also limited the number of F&V that their children tried. Mothers reported preferences for F&V are associated with estimates of their preschoolers' preferences for F&V. Relative to girls, boys may need additional opportunities for F&V exposure. Copyright © Taylor & Francis Group, LLC

Single-step colony assay for screening antibody libraries.

PubMed

Kato, Mieko; Hanyu, Yoshiro

2017-08-10

We describe a method, single-step colony assay, for simple and rapid screening of single-chain Fv fragment (scFv) libraries. Colonies of Escherichia coli expressing the scFv library are formed on a hydrophilic filter that is positioned in contact with a membrane coated with an antigen. scFv expression is triggered upon treatment of colonies with an induction reagent, following which scFvs are secreted from the cells and diffused to the antigen-coated membrane. scFvs that exhibit binding affinity for the antigen are captured by the membrane-immobilized antigen. Lastly, detection of scFv binding of the antigen on the membrane allows identification of the clones on the filter that express antigen-specific scFvs. We tested this methodology by using an anti-rabbit IgG scFv, scFv(A10B), and a rat immune scFv library. Experiments conducted using scFv(A10B) revealed that this method improves scFv expression during the colony assay. By using our method to screen an immune library of 3×10 3 scFv clones, we established several clones exhibiting affinity for the antigen. Moreover, we tested 7 other antigens, including peptides, and successfully identified positive clones. We believe that this simple procedure and controlled scFv expression of the single-step colony assay could make the antibody screening both rapid and reliable and lead to successful isolation of positive clones from antibody libraries. Copyright © 2017 Elsevier B.V. All rights reserved.

Differential stability of host mRNAs in Friend erythroleukemia cells infected with herpes simplex virus type 1

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

Mayman, B.A.; Nishioka, Y.

1985-01-01

The consequences of herpes simplex virus type 1 infection on cellular macromolecules were investigated in Friend erythroleukemia cells. The patterns of protein synthesis, examined by polyacrylamide gel electrophoresis, demonstrated that by 4 h postinfection the synthesis of many host proteins, with the exception of histones, was inhibited. Examination of the steady-state level of histone H3 mRNA by molecular hybridization of total RNA to a cloned mouse histone H3 complementary DNA probe demonstrated that the ratio of histone H3 mRNA to total RNA remained unchanged for the first 4 h postinfection. In contrast, the steady-state levels of globin and actin mRNAsmore » decreased progressively at early intervals postinfection. Studies on RNA synthesis in isolated nuclei demonstrated that the transcription of the histone H3 gene was inhibited to approximately the same extent as that of actin gene. It was concluded that the stabilization of preexisting histone H3 mRNA was responsible for the persistence of H3 mRNA and histone protein synthesis in herpes simplex virus type 1-infected Friend erythroleukemia cells. The possible mechanisms influencing the differential stability of host mRNAs during the course of productive infection with herpes simplex virus type 1 are discussed.« less

"Gay Boy Talk" Meets "Girl Talk": HIV Risk Assessment Assumptions in Young Gay Men's Sexual Health Communication with Best Friends

ERIC Educational Resources Information Center

Mutchler, Matt G.; McDavitt, Bryce

2011-01-01

Young adults, particularly young gay men (YGM), are vulnerable to human immunodeficiency virus (HIV). Yet, little is known about how YGM discuss sexual health issues with their friends ("gay boy talk"). We conducted semi-structured interviews with YGM and their best friends (11 YGM/YGM dyads and 13 YGM/heterosexual female dyads). In this paper, we…

Toward low-cost affinity reagents: lyophilized yeast-scFv probes specific for pathogen antigens.

PubMed

Gray, Sean A; Weigel, Kris M; Ali, Ibne K M; Lakey, Annie A; Capalungan, Jeremy; Domingo, Gonzalo J; Cangelosi, Gerard A

2012-01-01

The generation of affinity reagents, usually monoclonal antibodies, remains a critical bottleneck in biomedical research and diagnostic test development. Recombinant antibody-like proteins such as scFv have yet to replace traditional monoclonal antibodies in antigen detection applications, in large part because of poor performance of scFv in solution. To address this limitation, we have developed assays that use whole yeast cells expressing scFv on their surfaces (yeast-scFv) in place of soluble purified scFv or traditional monoclonal antibodies. In this study, a nonimmune library of human scFv displayed on the surfaces of yeast cells was screened for clones that bind to recombinant cyst proteins of Entamoeba histolytica, an enteric pathogen of humans. Selected yeast-scFv clones were stabilized by lyophilization and used in detection assay formats in which the yeast-scFv served as solid support-bound monoclonal antibodies. Specific binding of antigen to the yeast-scFv was detected by staining with rabbit polyclonal antibodies. In flow cytometry-based assays, lyophilized yeast-scFv reagents retained full binding activity and specificity for their cognate antigens after 4 weeks of storage at room temperature in the absence of desiccants or stabilizers. Because flow cytometry is not available to all potential assay users, an immunofluorescence assay was also developed that detects antigen with similar sensitivity and specificity. Antigen-specific whole-cell yeast-scFv reagents can be selected from nonimmune libraries in 2-3 weeks, produced in vast quantities, and packaged in lyophilized form for extended shelf life. Lyophilized yeast-scFv show promise as low cost, renewable alternatives to monoclonal antibodies for diagnosis and research.

No Time For Family Meals? Parenting Practices Associated With Adolescent Fruit And Vegetable Intake When Family Meals Are Not An Option

PubMed Central

Loth, Katie; Berge, Jerica M.; Larson, Nicole; Neumark-Sztainer, Dianne

2017-01-01

Background Despite research linking family meals to healthier diets, some families are unable to have regular meals together. These families need guidance about other ways to promote healthy eating among adolescents. Objective To examine the association between various parenting practices and adolescent fruit and vegetable (FV) intake at different levels of family meal frequency. Design Cross-sectional, population-based survey of influences on adolescent weight-related behaviors: EAT 2010 (Eating and Activity in Teens). Participants/Setting 2,491 adolescents recruited from middle/high schools in Minneapolis/St-Paul Measures Adolescent FV intake was ascertained with a food frequency questionnaire. Survey items assessed frequency of family meals and FV parenting practices (availability, accessibility, parent modeling, parent encouragement, and family communication). Statistical Analyses Linear regression models were used to examine associations with and interactions among family meals and parenting practices. Models were adjusted for age, sex, socioeconomic status, race/ethnicity, and energy intake (kcal/day). Results Family meals, FV availability, FV accessibility, FV modeling, and encouragement to eat healthy foods were independently associated with higher FV intake. Of the 949 (34%) adolescents who reported infrequent family meals (≤2 days/week), mean FV intake was 3.6 servings/day for those with high home FV availability versus 3.0 servings/day for those with low home FV availability. Similar differences in mean FV intake (0-3-0.6 servings/day) were found for high versus low FV accessibility, parental modeling, and parent encouragement for healthy eating. Frequent family meals in addition to more favorable parenting practices were associated with the highest FV intakes. Conclusion Food parenting practices and family meals are associated with greater adolescent FV intake. Longitudinal and intervention studies are needed to determine which combination of parenting practices will lead to improvements in adolescent diet. PMID:27989447

Fragaria vesca CONSTANS controls photoperiodic flowering and vegetative development.

PubMed

Kurokura, Takeshi; Samad, Samia; Koskela, Elli; Mouhu, Katriina; Hytönen, Timo

2017-10-13

According to the external coincidence model, photoperiodic flowering occurs when CONSTANS (CO) mRNA expression coincides with light in the afternoon of long days (LDs), leading to the activation of FLOWERING LOCUS T (FT). CO has evolved in Brassicaceae from other Group Ia CO-like (COL) proteins which do not control photoperiodic flowering in Arabidopsis. COLs in other species have evolved different functions as floral activators or even as repressors. To understand photoperiodic development in the perennial rosaceous model species woodland strawberry, we functionally characterized FvCO, the only Group Ia COL in its genome. We demonstrate that FvCO has a major role in the photoperiodic control of flowering and vegetative reproduction through runners. FvCO is needed to generate a bimodal rhythm of FvFT1 which encodes a floral activator in the LD accession Hawaii-4: a sharp FvCO expression peak at dawn is followed by the FvFT1 morning peak in LDs indicating possible direct regulation, but additional factors that may include FvGI and FvFKF1 are probably needed to schedule the second FvFT1 peak around dusk. These results demonstrate that although FvCO and FvFT1 play major roles in photoperiodic development, the CO-based external coincidence around dusk is not fully applicable to the woodland strawberry. © The Author 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Effects of ranavirus infection of red-eared sliders (Trachemys scripta elegans) on plasma proteins.

PubMed

Moore, A Russell; Allender, Matthew C; MacNeill, Amy L

2014-06-01

Ranavirus is an emerging disease that infects fish, amphibians, and reptiles. Ranavirus induces an inflammatory response leading to death in many susceptible species. Red-eared sliders (RES; Trachemys scripta elegans) are vulnerable to ranavirus infection and are economically significant chelonians kept in the pet trade and utilized in research. Early identification of RES with inflammatory diseases would allow for isolation of affected individuals and subsequent disease investigation, including molecular testing for ranavirus. Validation of an inexpensive, clinically relevant, and reproducible diagnostic test that detects inflammation in turtles is needed. Although commonly used, plasma protein electrophoresis to detect an inflammatory acute-phase protein response has not been evaluated in a controlled environment in turtles with experimentally induced inflammatory disease. The objective of this study was to measure plasma protein fractions by electrophoresis to determine if an acute-phase protein response occurs in RES during infection with a frog virus 3-like ranavirus (FV3-like virus) isolated from a chelonian. A Bradford assay and agarose gel electrophoresis (AGE) were performed using plasma collected during a study of the effect of temperature on the pathogenesis of ranavirus in RES. In RES at the time of viremia, total albumin (ALB(mg/ml)) and albumin to globulin ratio were significantly lower and beta-globulin percentage was significantly higher in RES exposed to ranavirus (n = 4) as compared to matched, uninfected RES (n = 8). In the last sample collected prior to death, total protein (TP(mg/ml)), ALB(mg/ml), alpha-globulin percentage, and total alpha-globulin (alpha(mg/ml)) were significantly lower in RES exposed to ranavirus (n = 4) than control individuals (n = 8). In summary, FV3-like virus induces a decrease in plasma albumin concentration at the onset ofviremia and decreases in TP(mg/ml, ALB(mg/ml), and alpha(mg/ml) concentrations prior to death in RES as measured by AGE.

Unique variations of Epstein-Barr virus-encoded BARF1 gene in nasopharyngeal carcinoma biopsies.

PubMed

Wang, Yun; Wang, Xiao-Feng; Sun, Zhi-Fu; Luo, Bing

2012-06-01

The Epstein-Barr virus (EBV) BamHI-A rightward frame 1 (BARF1) gene is frequently expressed in EBV-associated epithelial malignancies and involves in oncogenicity and immunomodulation. To characterize the variations of BARF1 gene in different populations, the sequences of BARF1 gene in Northern Chinese nasopharyngeal carcinoma (NPC), EBV-associated gastric carcinoma (EBVaGC) and healthy donors were analyzed. The correlation of BARF1 variation with polymorphisms of BamHI F fragment (type F and f variants) and EBV-coded viral interleukin-10 (vIL-10) gene (B95-8 and SPM patterns) was also explored. Two major subtypes of BARF1 gene, designated as B95-8 and V29A, were identified. B95-8 subtype had identical amino acid sequence to B95-8 and was the dominant subtype among the EBV isolates from Northern China. V29A subtype, with one consistent amino acid change at residue 29 (V→A) and several nucleotide changes, showed higher frequency in NPC cases (25.3%, 20/79) than in EBVaGC cases (0/45) or healthy donors (4.3%, 2/46) (NPC vs. EBVaGC: P=0.0001; NPC vs. healthy donor: P=0.004). A preferential linkage between BamHI F and BARF1/vIL-10 polymorphisms was found. Type f isolates was specially correlated with the V29A/SPM genotype in NPC isolates and type f/V29A/SPM was preferentially found in NPC. BARF1/c-fms homology domain, transforming domain and cytotoxic T lymphocyte (CTL) epitopes of BARF1 were highly conserved in most isolates, suggesting the important role of BARF1 in virus infection and the potential usefulness in EBV-targeting immunotherapy of EBV-associated tumors. The relatively higher prevalence of type f/V29A/SPM strains in NPC may also suggest the association between these variations in multiple viral genes and NPC. Copyright © 2012 Elsevier B.V. All rights reserved.

77 FR 35850 - Safety Zone; F/V Deep Sea, Penn Cove, WA

Federal Register 2010, 2011, 2012, 2013, 2014

2012-06-15

... 1625-AA00 Safety Zone; F/V Deep Sea, Penn Cove, WA AGENCY: Coast Guard, DHS. ACTION: Temporary final rule. SUMMARY: The Coast Guard is establishing a safety zone around the Fishing Vessel (F/V) Deep Sea... with the sunken F/V Deep Sea. B. Basis and Purpose On the evening of May 13, 2012, the F/V Deep Sea...

Efficacy of solvent/detergent plasma after storage at 2-8 °C for 5 days in comparison to other plasma products to improve factor V levels in factor V deficient plasma.

PubMed

Cushing, Melissa M; Asmis, Lars; Calabia, Carmencita; Rand, Jacob H; Haas, Thorsten

2016-08-01

Factor V (FV) plays an important role in coagulation. As no purified concentrate is available to restore critical FV levels, the main blood product used to replace FV is plasma. The aim of the present in vitro study was to compare the efficacy of the different available plasma products on the reversal of moderate and severe FV deficiency as assessed by ROTEM® and FV levels. Five different plasma products (6 batches of each) were compared to determine their effectiveness in replacing FV in plasma moderately or severely deficient in FV. Effectiveness was measured using the ROTEM® EXTEM clotting time (CT) and a factor V assay. FFP, plasma frozen within 24 hours (FP24), Octaplas (solvent/detergent treated pooled plasma), as well as Octaplas and FP24 thawed and stored for 5 days (Octaplas TP and TP), were all used for in vitro replacement of FV. TP was significantly less effective at reversing a prolonged EXTEM CT and FV levels in FV deficient plasma than other tested products. There were no significant differences in EXTEM CT between Octaplas and Octaplas TP, while factor V activity was significantly lower in the Octaplas TP. There was no significant difference between Octaplas and FFP for EXTEM CT or FV activity. Octaplas and Octaplas TP appear to have an equivalent ability to improve the EXTEM CT and could be considered as a treatment alternative to FFP in patients with FV deficiency. Copyright © 2016 Elsevier Ltd. All rights reserved.

The Role of B Cells for in Vivo T Cell Responses to a Friend Virus-Induced Leukemia

NASA Astrophysics Data System (ADS)

Schultz, Kirk R.; Klarnet, Jay P.; Gieni, Randall S.; Hayglass, Kent T.; Greenberg, Philip D.

1990-08-01

B cells can function as antigen-presenting cells and accessory cells for T cell responses. This study evaluated the role of B cells in the induction of protective T cell immunity to a Friend murine leukemia virus (F-MuLV)-induced leukemia (FBL). B cell-deficient mice exhibited significantly reduced tumor-specific CD4^+ helper and CD8^+ cytotoxic T cell responses after priming with FBL or a recombinant vaccinia virus containing F-MuLV antigens. Moreover, these mice had diminished T cell responses to the vaccinia viral antigens. Tumor-primed T cells transferred into B cell-deficient mice effectively eradicated disseminated FBL. Thus, B cells appear necessary for efficient priming but not expression of tumor and viral T cell immunity.

Engineered Recombinant Single-Chain Fragment Variable Antibody for Immunosensors

PubMed Central

Shen, Zhihong; Mernaugh, Raymond L.; Yan, Heping; Yu, Lei; Zhang, Ying; Zeng, Xiangqun

2008-01-01

A recombinant single-chain fragment variable (scFv) antibody (designated A10B) was engineered to contain two histidines within the linker peptide used to join the scFv heavy and light chains. A piezoimmunosensor using the scFv was successfully developed. A10B scFv bound to the gold piezoimmunosensor surface were correctly oriented, retained antigen-binding activity, and coupled at high surface concentration. These results, and results obtained from an earlier study using an scFv containing a linker cysteine, suggest that the location on the linker sequence in which the amino acids were incorporated was well tolerated by the scFv and did not interfere with scFv antigen-binding activity. The scFv-modified QCM sensor was thoroughly characterized and used to specifically detect antigen in crude serum sample and had a sensitivity of 2.3 ± 0.15 nM (n = 4) with a linear range over 2.3 × 10−9–3.3 × 10−8 M. The piezoimmunosensor was also used to study the kinetics and thermodynamics of antigen/scFv antibody binding. PMID:16255580

The Fragaria vesca Homolog of SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 Represses Flowering and Promotes Vegetative Growth[W

PubMed Central

Mouhu, Katriina; Kurokura, Takeshi; Koskela, Elli A.; Albert, Victor A.; Elomaa, Paula; Hytönen, Timo

2013-01-01

In the annual long-day plant Arabidopsis thaliana, SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (SOC1) integrates endogenous and environmental signals to promote flowering. We analyzed the function and regulation of the SOC1 homolog (Fragaria vesca [Fv] SOC1) in the perennial short-day plant woodland strawberry (Fragaria vesca). We found that Fv SOC1 overexpression represses flower initiation under inductive short days, whereas its silencing causes continuous flowering in both short days and noninductive long days, similar to mutants in the floral repressor Fv TERMINAL FLOWER1 (Fv TFL1). Molecular analysis of these transgenic lines revealed that Fv SOC1 activates Fv TFL1 in the shoot apex, leading to the repression of flowering in strawberry. In parallel, Fv SOC1 regulates the differentiation of axillary buds to runners or axillary leaf rosettes, probably through the activation of gibberellin biosynthetic genes. We also demonstrated that Fv SOC1 is regulated by photoperiod and Fv FLOWERING LOCUS T1, suggesting that it plays a central role in the photoperiodic control of both generative and vegetative growth in strawberry. In conclusion, we propose that Fv SOC1 is a signaling hub that regulates yearly cycles of vegetative and generative development through separate genetic pathways. PMID:24038650

Crystal Structure and Size-Dependent Neutralization Properties of HK20, a Human Monoclonal Antibody Binding to the Highly Conserved Heptad Repeat 1 of gp41

PubMed Central

Seaman, Mike S.; Lutje Hulsik, David; Hinz, Andreas; Vanzetta, Fabrizia; Agatic, Gloria; Silacci, Chiara; Mainetti, Lara; Scarlatti, Gabriella; Sallusto, Federica; Weiss, Robin; Lanzavecchia, Antonio; Weissenhorn, Winfried

2010-01-01

The human monoclonal antibody (mAb) HK20 neutralizes a broad spectrum of primary HIV-1 isolates by targeting the highly conserved heptad repeat 1 (HR1) of gp41, which is transiently exposed during HIV-1 entry. Here we present the crystal structure of the HK20 Fab in complex with a gp41 mimetic 5-Helix at 2.3 Å resolution. HK20 employs its heavy chain CDR H2 and H3 loops to bind into a conserved hydrophobic HR1 pocket that is occupied by HR2 residues in the gp41 post fusion conformation. Compared to the previously described HR1-specific mAb D5, HK20 approaches its epitope with a different angle which might favor epitope access and thus contribute to its higher neutralization breadth and potency. Comparison of the neutralization activities of HK20 IgG, Fab and scFv employing both single cycle and multiple cycle neutralization assays revealed much higher potencies for the smaller Fab and scFv over IgG, implying that the target site is difficult to access for complete antibodies. Nevertheless, two thirds of sera from HIV-1 infected individuals contain significant titers of HK20-inhibiting antibodies. The breadth of neutralization of primary isolates across all clades, the higher potencies for C-clade viruses and the targeting of a distinct site as compared to the fusion inhibitor T-20 demonstrate the potential of HK20 scFv as a therapeutic tool. PMID:21124990

Crystal structure and size-dependent neutralization properties of HK20, a human monoclonal antibody binding to the highly conserved heptad repeat 1 of gp41.

PubMed

Sabin, Charles; Corti, Davide; Buzon, Victor; Seaman, Mike S; Lutje Hulsik, David; Hinz, Andreas; Vanzetta, Fabrizia; Agatic, Gloria; Silacci, Chiara; Mainetti, Lara; Scarlatti, Gabriella; Sallusto, Federica; Weiss, Robin; Lanzavecchia, Antonio; Weissenhorn, Winfried

2010-11-18

The human monoclonal antibody (mAb) HK20 neutralizes a broad spectrum of primary HIV-1 isolates by targeting the highly conserved heptad repeat 1 (HR1) of gp41, which is transiently exposed during HIV-1 entry. Here we present the crystal structure of the HK20 Fab in complex with a gp41 mimetic 5-Helix at 2.3 Å resolution. HK20 employs its heavy chain CDR H2 and H3 loops to bind into a conserved hydrophobic HR1 pocket that is occupied by HR2 residues in the gp41 post fusion conformation. Compared to the previously described HR1-specific mAb D5, HK20 approaches its epitope with a different angle which might favor epitope access and thus contribute to its higher neutralization breadth and potency. Comparison of the neutralization activities of HK20 IgG, Fab and scFv employing both single cycle and multiple cycle neutralization assays revealed much higher potencies for the smaller Fab and scFv over IgG, implying that the target site is difficult to access for complete antibodies. Nevertheless, two thirds of sera from HIV-1 infected individuals contain significant titers of HK20-inhibiting antibodies. The breadth of neutralization of primary isolates across all clades, the higher potencies for C-clade viruses and the targeting of a distinct site as compared to the fusion inhibitor T-20 demonstrate the potential of HK20 scFv as a therapeutic tool.

Selection of stable scFv antibodies by phage display.

PubMed

Brockmann, Eeva-Christine

2012-01-01

ScFv fragments are popular recombinant antibody formats but often suffer from limited stability. Phage display is a powerful tool in antibody engineering and applicable also for stability selection. ScFv variants with improved stability can be selected from large randomly mutated phage displayed libraries with a specific antigen after the unstable variants have been inactivated by heat or GdmCl. Irreversible scFv denaturation, which is a prerequisite for efficient selection, is achieved by combining denaturation with reduction of the intradomain disulfide bonds. Repeated selection cycles of increasing stringency result in enrichment of stabilized scFv fragments. Procedures for constructing a randomly mutated scFv library by error-prone PCR and phage display selection for enrichment of stable scFv antibodies from the library are described here.

Know Your Enemy: Successful Bioinformatic Approaches to Predict Functional RNA Structures in Viral RNAs.

PubMed

Lim, Chun Shen; Brown, Chris M

2017-01-01

Structured RNA elements may control virus replication, transcription and translation, and their distinct features are being exploited by novel antiviral strategies. Viral RNA elements continue to be discovered using combinations of experimental and computational analyses. However, the wealth of sequence data, notably from deep viral RNA sequencing, viromes, and metagenomes, necessitates computational approaches being used as an essential discovery tool. In this review, we describe practical approaches being used to discover functional RNA elements in viral genomes. In addition to success stories in new and emerging viruses, these approaches have revealed some surprising new features of well-studied viruses e.g., human immunodeficiency virus, hepatitis C virus, influenza, and dengue viruses. Some notable discoveries were facilitated by new comparative analyses of diverse viral genome alignments. Importantly, comparative approaches for finding RNA elements embedded in coding and non-coding regions differ. With the exponential growth of computer power we have progressed from stem-loop prediction on single sequences to cutting edge 3D prediction, and from command line to user friendly web interfaces. Despite these advances, many powerful, user friendly prediction tools and resources are underutilized by the virology community.

Know Your Enemy: Successful Bioinformatic Approaches to Predict Functional RNA Structures in Viral RNAs

PubMed Central

Lim, Chun Shen; Brown, Chris M.

2018-01-01

Structured RNA elements may control virus replication, transcription and translation, and their distinct features are being exploited by novel antiviral strategies. Viral RNA elements continue to be discovered using combinations of experimental and computational analyses. However, the wealth of sequence data, notably from deep viral RNA sequencing, viromes, and metagenomes, necessitates computational approaches being used as an essential discovery tool. In this review, we describe practical approaches being used to discover functional RNA elements in viral genomes. In addition to success stories in new and emerging viruses, these approaches have revealed some surprising new features of well-studied viruses e.g., human immunodeficiency virus, hepatitis C virus, influenza, and dengue viruses. Some notable discoveries were facilitated by new comparative analyses of diverse viral genome alignments. Importantly, comparative approaches for finding RNA elements embedded in coding and non-coding regions differ. With the exponential growth of computer power we have progressed from stem-loop prediction on single sequences to cutting edge 3D prediction, and from command line to user friendly web interfaces. Despite these advances, many powerful, user friendly prediction tools and resources are underutilized by the virology community. PMID:29354101

[Construction of a phage antibody library and screening of anti-epidermal growth factor receptor variant III single chain antibody].

PubMed

Han, Dong-gang; Duan, Xiao-yi; Guo, You-min; Zhou, Qi; Wang, Quan-ying; Yang, Guang-xiao

2010-01-01

To obtain specific anti-epidermal growth factor receptor variant III (EGFRvIII) single chain antibody (ScFv) by phage antibody library display system. The total RNA was extracted from the spleen B cells of BALB/c mice immunized with pep-3-OVA protein, and the first-strand cDNA was synthesized by reverse transcription. Antibody VH and VL gene fragments were amplified and joined to a ScFv gene with the linker. The ScFv gene was ligated into the phagemid vector pCANTAB5E, which was transformed into competent E. coli TG1. The transformed cells were then infected with M13KO7 helper phage to yield the recombinant phage to construct the phage ScFv library. Pep-3-BSA protein was used to screen the phage antibody library and ELISA carried out to characterize the activity of the antibody. The VH and VL gene fragments of the antibody were about 350 bp and 320 bp in length as analyzed by agarose gel electrophoresis. The ScFv gene was 780 bp, consistent with the expected length. The recombinant phagemid with ScFv gene insert was rescued, and an immune phage ScFv library with the content of 5.0x10(6) was constructed. The recombinant ScFv phage had a titer of 3.0x10(4) cfu/ml, and the fourth phage harvest yielded 56 times as much as that of the first one. SDS-PAGE demonstrated a molecular mass of the soluble ScFv of about 28 kD. ELISA results indicated good specificity of the ScFv to bind EGFRvIII. An immune phage ScFv library is successfully constructed, and the ScFv antibody fragment is capable of specific binding to EGFRvIII.

No Time for Family Meals? Parenting Practices Associated with Adolescent Fruit and Vegetable Intake When Family Meals Are Not an Option.

PubMed

Watts, Allison W; Loth, Katie; Berge, Jerica M; Larson, Nicole; Neumark-Sztainer, Dianne

2017-05-01

Despite research linking family meals to healthier diets, some families are unable to have regular meals together. These families need guidance about other ways to promote healthy eating among adolescents. Our aim was to examine the association between various parenting practices and adolescent fruit and vegetable (F/V) intake at different levels of family meal frequency. We conducted a cross-sectional, population-based survey of influences on adolescent weight-related behaviors using Project EAT (Eating and Activity in Teens) 2010. Participants were 2,491 adolescents recruited from middle/high schools in Minneapolis/St Paul, MN. Adolescent F/V intake was ascertained with a food frequency questionnaire. Survey items assessed frequency of family meals and F/V parenting practices (availability, accessibility, parent modeling, parent encouragement, and family communication). Linear regression models were used to examine associations with and interactions among family meals and parenting practices. Models were adjusted for age, sex, socioeconomic status, race/ethnicity, and energy intake (kilocalories per day). Family meals, F/V availability, F/V accessibility, F/V modeling, and encouragement to eat healthy foods were independently associated with higher F/V intake. Of the 949 (34%) adolescents who reported infrequent family meals (≤2 days/wk), mean F/V intake was 3.6 servings/day for those with high home F/V availability vs 3.0 servings/day for those with low home F/V availability. Similar differences in mean F/V intake (0.3 to 0.6 servings/day) were found for high vs low F/V accessibility, parental modeling, and parent encouragement for healthy eating. Frequent family meals in addition to more favorable parenting practices were associated with the highest F/V intakes. Food parenting practices and family meals are associated with greater adolescent F/V intake. Longitudinal and intervention studies are needed to determine which combination of parenting practices will lead to improvements in adolescent diets. Copyright © 2017 Academy of Nutrition and Dietetics. Published by Elsevier Inc. All rights reserved.

Symmetry of Fv architecture is conducive to grafting a second antibody binding site in the Fv region.

PubMed Central

Keck, P C; Huston, J S

1996-01-01

Molecular modeling studies on antibody Fv regions have been pursued to design a second antigen-binding site (chi-site) in a chimeric single-chain Fv (chi sFv) species of about 30 kDa. This analysis has uncovered an architectural basis common to many Fv regions that permits grafting a chi-site onto the Fv surface that diametrically opposes the normal combining site. By using molecular graphics analysis, chimeric complementarity-determining regions (chi CDRs) were defined that comprised most of the CDRs from an antibody binding site of interest. The chain directionality of chi CDRs was consistent with that of specific bottom loops of the sFv, which allowed for grafting of chi CDRs with an overall geometry approximating CDRs in the parent combining site. Analysis of 10 different Fv crystal structures indicates that the positions for inserting chi CDRs are very highly conserved, as are the corresponding chi CDR boundaries in the parent binding site. The results of this investigation suggest that it should be possible to generally apply this approach to the development of chimeric bispecific antibody binding site (chi BABS) proteins. Images FIGURE 2 FIGURE 3 PMID:8889174

Novel avian single-chain fragment variable (scFv) targets dietary gluten and related natural grain prolamins, toxic entities of celiac disease.

PubMed

Stadlmann, Valerie; Harant, Hanna; Korschineck, Irina; Hermann, Marcela; Forster, Florian; Missbichler, Albert

2015-12-01

Celiac disease (CD) is a chronic, small intestinal inflammatory disease mediated by dietary gluten and related prolamins. The only current therapeutic option is maintenance of a strict life-long gluten-free diet, which implies substantial burden for CD patients. Different treatment regimes might be feasible, including masking of toxic celiac peptides with blocking antibodies or fragments thereof. The objective of this study was therefore to select and produce a recombinant avian single-chain fragment variable (scFv) directed against peptic-tryptic digested gliadin (PT-Gliadin) and related celiac toxic entities. Gluten-free raised chicken of same age were immunized with PT-Gliadin. Chicken splenic lymphocytes, selected with antigen-coated magnetic beads, served as RNA source for the generation of cDNA. Chicken VH and VL genes were amplified from the cDNA by PCR to generate full-length scFv constructs consisting of VH and VL fragments joined by a linker sequence. ScFv constructs were ligated in a prokaryotic expression vector, which provides a C-terminal hexahistidine tag. ScFvs from several bacterial clones were expressed in soluble form and crude cell lysates screened for binding to PT-Gliadin by ELISA. We identified an enriched scFv motif, which showed reactivity to PT-Gliadin. One selected scFv candidate was expressed and purified to homogeneity. Polyclonal anti-PT-Gliadin IgY, purified from egg yolk of immunized chicken, served as control. ScFv binds in a dose-dependent manner to PT-Gliadin, comparable to IgY. Furthermore, IgY competitively displaces scFv from PT-Gliadin and natural wheat flour digest, indicating a common epitope of scFv and IgY. ScFv was tested for reactivity to different gastric digested dietary grain flours. ScFv detects common and khorasan wheat comparably with binding affinities in the high nanomolar range, while rye is detected to a lesser extent. Notably, barley and cereals which are part of the gluten-free diet, like corn and rice, are not detected by scFv. Similarly, the pseudo-grain amaranth, used as gluten-free alternative, is not targeted by scFv. This data indicate that scFv specifically recognizes toxic cereal peptides relevant in CD. ScFv can be of benefit for future CD treatment regimes.

PET Imaging of 64Cu-DOTA-scFv-Anti-PSMA Lipid Nanoparticles (LNPs): Enhanced Tumor Targeting over Anti-PSMA scFv or Untargeted LNPs

PubMed Central

Wong, Patty; Li, Lin; Chea, Junie; Delgado, Melissa K.; Crow, Desiree; Poku, Erasmus; Szpikowska, Barbara; Bowles, Nicole; Channappa, Divya; Colcher, David; Wong, Jeffrey Y.C.; Shively, John E.; Yazaki, Paul J.

2017-01-01

Introduction Single chain (scFv) antibodies are ideal targeting ligands due to their modular structure, high antigen specificity and affinity. These monovalent ligands display rapid tumor targeting but have limitations due to their fast urinary clearance. Methods An anti-prostate membrane antigen (PSMA) scFv with a site-specific cysteine was expressed and evaluated in a prostate cancer xenograft model by Cu-64 PET imaging. To enhance tumor accumulation, the scFv-cys was conjugated to the co-polymer DSPE-PEG-maleimide that spontaneously assembled into a homogeneous multivalent lipid nanoparticle (LNP). Results The targeted LNP exhibited a 2-fold increase in tumor uptake compared to the scFv alone using two different thiol ester chemistries. The anti-PSMA scFv-LNP exhibited a 1.6 fold increase in tumor targeting over the untargeted LNP. Conclusions The targeted anti-PSMA scFv-LNP showed enhanced tumor accumulation over the scFv alone or the untargeted DOTA-micelle providing evidence for the development of this system for drug delivery. Advances in Knowledge and implications for patient care Anti-tumor scFv antibody fragments have not achieved their therapeutic potential due to their fast blood clearance. Conjugation to a LNP enables multivalency to the tumor antigen as well as increased molecular size for chemotherapy drug delivery. PMID:28126683

Production of a germline-humanized cetuximab scFv and evaluation of its activity in recognizing EGFR- overexpressing cancer cells.

PubMed

Banisadr, Arsham; Safdari, Yaghoub; Kianmehr, Anvarsadat; Pourafshar, Mahdieh

2018-04-03

The aim of this study was to produce a humanized single chain antibody (scFv) as a potential improved product design to target EGFR (Epidermal Growth Factor Receptor) overexpressing cancer cells. To this end, CDR loops of cetuximab (an FDA-approved anti-EGFR antibody) were grafted on framework regions derived from type 3 (VH3 and VL3 kappa) human germline sequences to obtain recombinant VH and VL domainslinked together with a flexible linker [(Gly 4 Ser) 3 ] to form a scFv. Codon optimized synthetic gene encoding the scFv (with NH2-VH-linker-VL-COOH orientation) was expressed in E. coli Origami™ 2(DE3) cells and the resultant scFv purified by using Ni-NTA affinity chromatography. The scFv, called cet.Hum scFv, was evaluated in ELISA and immunoblot to determine whether it can recognize EGFR. The scFv was able to recognize EGFR over-expressing cancer cells (A-431) but failed to detect cancer cells with low levels of EGFR (MCF-7 cells). Although the affinity of the scFv forA-431 cells was 9 fold lower than that of cetuximab, it was strong enough to recognize these cells. Considering its ability to bind EGFR molecules, the scFv may exhibit a potential application for the detection of EGFR-overexpressing cancer cells.

Production of Recombinant Human scFv Against Tetanus Toxin Heavy Chain by Phage Display Technology.

PubMed

Khalili, Ehsan; Lakzaei, Mostafa; Rasaee, Mohhamad Javad; Aminian, Mahdi

2015-10-01

Tetanus, as a major cause of death in developing countries, is caused by tetanus neurotoxin. Recombinant antibodies against tetanus neurotoxin can be useful in tetanus management. Phage display of antibody fragments from immune human antibody libraries with single chain constructs combining the variable fragments (scFv) has been one of the most prominent technologies in antibody engineering. The aim of this study was the generation of a single chain fragment of variable region (scFv) library and selection of specific antibodies with high affinity against tetanus toxin. Immune human single chain fragment variable (HuscFv) antibody phagemid library was displayed on pIII of filamentous bacteriophage. Selection of scFv clones was performed against tetanus toxin antigens after three rounds of panning. The selected scFv clones were analyzed for inhibition of tetanus toxin binding to ganglioside GT1b. After the third round of panning, over 35 HuscFv phages specific for tetanus toxin were isolated from this library of which 15 clones were found to bind specifically to tetanus toxin. The selected HuscFv phages expressed as a soluble HuscFv peptide and some clones showed positive signals against tetanus toxin. We found that six HuscFv clones inhibit toxin binding to ganglioside GT1b. These selected antibodies can be used in the management of tetanus.

Inverse Association between Fruit and Vegetable Intake and BMI even after Controlling for Demographic, Socioeconomic and Lifestyle Factors

PubMed Central

Heo, Moonseong; Kim, Ryung S.; Wylie-Rosett, Judith; Allison, David B.; Heymsfield, Steve B.; Faith, Myles S.

2011-01-01

Objective To estimate fruit and vegetable (FV) intake levels of US adult population and evaluate the association between FV intake and BMI status after controlling for confounding demographic, socioeconomic and lifestyle factors. We also sought to identify moderating factors. Methods We used 2007 Behavior Risk Factors Surveillance System (N > 400,000) data. FV intake was dichotomized as ≥5 servings (FV5+) versus <5 servings/day. BMI status was categorized as normal, overweight, and obese. Identification of moderators was performed by testing interactions between BMI status and other variables using bivariate analyses followed by multiple logistic regression analysis incorporating complex survey sampling design features. Results Only 24.6% of US adults consumed ≥5 servings per day and less than 4% consumed 9 or more servings. Overweight (% FV5+ = 23.9%) and obese (21.9%) groups consumed significantly less FV than the normal-weight (27.4%) group (p < 0.0001). This inverse association remained significant even after controlling for potential confounding factors. Multivariate analysis identified five significant moderators (p < 0.0001) after controlling for all evaluated variables: race, sex, smoking status, health coverage, and physical activity. Notably, physically inactive obese males tended to consume the least FV (% FV5+ = 14.7%). Conclusion Current US population FV intake level is below recommended levels. The inverse association between FV intake and obesity was significant and was moderated by demographic, socioeconomic status, and lifestyle factors. These factors should be considered when developing policies and interventions to increase FV intake. PMID:22248995

Fruit and vegetable attitudes, norms, and intake in low-income youth

USDA-ARS?s Scientific Manuscript database

Fruit and vegetable (FV) attitudes and norms have been shown to influence intake in youth; yet research with low-income youth and studies supplementing self-report with objective measures of intake are lacking. Cross-sectional survey data on self-rated FV intake, FV attitudes, and FV norms were coll...

Provision of Fluoride Varnish to Medicaid-Enrolled Children by Physicians: The Massachusetts Experience

PubMed Central

Isong, Inyang A; Silk, Hugh; Rao, Sowmya R; Perrin, James M; Savageau, Judith A; Donelan, Karen

2011-01-01

Objectives To evaluate the impact of a 2008 Medicaid policy in Massachusetts (MA), regarding reimbursing physicians for providing fluoride varnish (FV) to eligible children in medical settings. Data Source Survey of a sample of primary care physicians in MA. Study Design Cross-sectional survey of a sample of physicians who provide care to MassHealth (MA Medicaid) enrolled-children. Dependent variables: history of completed preventive dental skills training, and FV provision. Independent variables: oral health knowledge, FV-attitudes, and physician and practice characteristics. Principal Findings Overall, 19 percent of respondents had completed the training required to be eligible to bill for FV provision. Only 5 percent of physicians were providing FV. Most respondents (63 percent) were not familiar with the new policy, and only 25 percent felt that FV should be provided during well-child visits. Most physicians (60 percent) did not feel that the reimbursement rate of U.S.$26/application was sufficient; 17 percent said that they would not provide FV, regardless of payment. Most common barriers to FV provision were a lack of time and logistical challenges. Conclusions Our findings suggest that simply reimbursing physicians for FV provision is insufficient to ensure provider participation. Success of this policy will likely require addressing several barriers identified. PMID:21762142

Efficacy and safety of a clinically relevant foamy vector design in human hematopoietic repopulating cells.

PubMed

Everson, Elizabeth M; Hocum, Jonah D; Trobridge, Grant D

2018-06-23

Previous studies have shown that foamy viral (FV) vectors are a promising alternative to gammaretroviral and lentiviral vectors and insulators can improve FV vector safety. However, in a previous analysis of insulator effects on FV vector safety, strong viral promoters were used to elicit genotoxic events. Here we developed and analyzed the efficacy and safety of a high-titer, clinically relevant FV vector driven by the housekeeping promoter elongation factor-1α and insulated with an enhancer blocking A1 insulator (FV-EGW-A1). Human CD34 + cord blood cells were exposed to an enhanced green fluorescent protein expressing vector, FV-EGW-A1, at a multiplicity of infection of 10 and then maintained in vitro or transplanted into immunodeficient mice. Flow cytometry was used to measure engraftment and marking in vivo. FV vector integration sites were analyzed to assess safety. FV-EGW-A1 resulted in high-marking, multi-lineage engraftment of human repopulating cells with no evidence of silencing. Engraftment was highly polyclonal with no clonal dominance and a promising safety profile based on integration site analysis. An FV vector with an elongation factor-1α promoter and an A1 insulator is a promising vector design for use in the clinic. This article is protected by copyright. All rights reserved.

Bacterial Expression of a Single-Chain Variable Fragment (scFv) Antibody against Ganoderic Acid A: A Cost-Effective Approach for Quantitative Analysis Using the scFv-Based Enzyme-Linked Immunosorbent Assay.

PubMed

Yusakul, Gorawit; Nuntawong, Poomraphie; Sakamoto, Seiichi; Ratnatilaka Na Bhuket, Pahweenvaj; Kohno, Toshitaka; Kikkawa, Nao; Rojsitthisak, Pornchai; Shimizu, Kuniyoshi; Tanaka, Hiroyuki; Morimoto, Satoshi

2017-01-01

Due to the highly specific binding between an antibody and its target, superior analytical performances was obtained by immunoassays for phytochemical analysis over conventional chromatographic techniques. Here, we describe a simple method for producing a functional single-chain variable fragment (scFv) antibody against ganoderic acid A (GAA), a pharmacologically active metabolite from Ganoderma lingzhi. The Escherichia coli BL21(DE3) strain produced a large amount of anti-GAA scFv. However, in vitro refolding steps, which partially recovered the reactivity of the scFv, were required. Interestingly, the functional scFv was expressed as a soluble and active form in the cytoplasm of an engineered E. coli SHuffle ® strain. Purified anti-GAA scFv, which yielded 2.56 mg from 1 L of culture medium, was obtained from simple and inexpensive procedures for expression and purification. The anti-GAA scFv-based indirect competitive enzyme-linked immunosorbent assay (icELISA) exhibited high sensitivity (linearity: 0.078-1.25 µg/mL) with precision (CV: ≤6.20%) and reliability (recovery: 100.1-101.8%) for GAA determination. In summary, the approach described here is an inexpensive, simple, and efficient expression system that extends the application of anti-GAA scFv-based immunoassays. In addition, when in vitro refolding steps can be skipped, the cost and complexity of scFv antibody production can be minimized.

Isolation and characterization of a novel human scFv inhibiting EGFR vIII expressing cancers.

PubMed

Rahbarnia, Leila; Farajnia, Safar; Babaei, Hossein; Majidi, Jafar; Dariushnejad, Hassan; Hosseini, Mohammad Kazem

2016-12-01

EGFRvIII, a mutant form of epidermal growth factor receptor is highly expressed in glioblastoma, carcinoma of the breast, ovary, and lung but not in normal cells. This tumor specific antigen has emerged as a promising candidate for antibody based therapy of several cancers. The aim of the present study was isolation and characterization of a human single chain antibody against EGFRvIII as a promising target for cancer therapy. For this, a synthetic peptide corresponding to EGFRvIII protein was used for screening the naive human scFv phage library. Selection was performed using a novel screening strategy for enrichment of rare specific clones. After five rounds of screening, six positive scFv clones against EGFRvIII were selected using monoclonal phage ELISA, among them, a clone with an amber mutation in VH CDR2 coding sequence showed higher reactivity. The mutation was corrected through site directed mutagenesis and then scFv fragment was expressed after subcloning into the bacterial expression vector. Expression in BL21 pLysS resulted in a highly soluble scFv appeared in soluble fraction of E. coli lysate. Bioinformatic in silico analysis between scFv and EGFRvIII sequences confirmed specific binding of desired scFv to EGFRvIII in CDR regions. The specific reactivity of the purified scFv with native EGFRvIII was confirmed by cell based ELISA and western blot. In conclusion, human anti- EGFRvIII scFv isolated from a scFv phage library displayed high reactivity with EGFRvIII. The scFv isolated in this study can be the groundwork for developing more effective diagnostic and therapeutic agents against EGFRvIII expressing cancers. Copyright © 2016 European Federation of Immunological Societies. Published by Elsevier B.V. All rights reserved.

Consumption of fruits and vegetables among adolescents: a multi-national comparison of eleven countries in the Eastern Mediterranean Region.

PubMed

Al Ani, M F; Al Subhi, L K; Bose, S

2016-03-28

Regional cross-country profile of fruit and vegetable (F&V) consumption is lacking in the Eastern Mediterranean Region (EMR). This study examines the prevalence and differences of consuming F&V ≥5 times/d among adolescents in eleven EMR countries, and also describes differences in the proportions of taking F&V ≥5 times/d by sex, age and BMI. The study included 26 328 school adolescents (13-15 years) with complete data on consumption of F&V, age, sex, weight and height taken from the Global School-based Student Health Survey conducted in the EMR between 2005 and 2009. Overall, only 19·4 % of adolescents reported consuming F&V ≥5 times/d. The highest prevalence was reported in Djibouti (40·4 %) and the lowest was reported in Pakistan (10·0 %). Statistically significant differences in prevalence were observed across countries (P<0·05). With the exception of Oman, Libya and Djibouti, significantly more males than females ate F&V ≥5 times/d. Proportion of students consuming F&V ≥5 times/d also varied significantly in all counties based on BMI (P<0·0001), with students within normal BMI having the highest frequency. A negative trend was observed between age and the prevalence of taking F&V ≥5 times/d in most of the eleven EMR countries but Jordan, Djibouti and Morocco. The prevalence of adequate intake of F&V was low in the eleven EMR countries. There is a need for interventions to increase the prevalence of adolescents consuming F&V ≥5 times/d. Interventions should take into consideration psychosocial, environmental and socio-environmental factors influencing F&V intake within countries.

A simple and robust approach to immobilization of antibody fragments.

PubMed

Ikonomova, Svetlana P; He, Ziming; Karlsson, Amy J

2016-08-01

Antibody fragments, such as the single-chain variable fragment (scFv), have much potential in research and diagnostics because of their antigen-binding ability similar to a full-sized antibody and their ease of production in microorganisms. Some applications of antibody fragments require immobilization on a surface, and we have established a simple immobilization method that is based on the biotin-streptavidin interaction and does not require a separate purification step. We genetically fused two biotinylation tags-the biotin carboxyl carrier protein (BCCP) or the AviTag minimal sequence-to six different scFvs (scFv13R4, scFvD10, scFv26-10, scFv3, scFv5, and scFv12) for site-specific biotinylation in vivo by endogenous biotin ligases produced by Escherichia coli. The biotinylated scFvs were immobilized onto streptavidin-coated plates directly from cell lysates, and immobilization was detected through enzyme-linked immunosorbent assays. All scFvs fusions were successfully immobilized, and scFvs biotinylated via the BCCP tag tended to immobilize better than those biotinylated via the AviTag, even when biotinylation efficiency was improved with the biotin ligase BirA. The ability of immobilized scFvs to bind antigens was confirmed using scFv13R4 and scFvD10 with their respective targets β-galactosidase and bacteriophage lambda head protein D (gpD). The immobilized scFv13R4 bound to β-galactosidase at the same level for both biotinylation tags when the surface was saturated with the scFv, and immobilized scFvs retained their functionality for at least 100days after immobilization. The simplicity and robustness of our method make it a promising approach for future applications that require antibody fragment immobilization. Copyright © 2016 Elsevier B.V. All rights reserved.

A strategy for high-level expression of a single-chain variable fragment against TNFα by subcloning antibody variable regions from the phage display vector pCANTAB 5E into pBV220.

PubMed

Yang, Tao; Yang, Lijun; Chai, Weiran; Li, Renke; Xie, Jun; Niu, Bo

2011-03-01

A phage display single-chain variable fragment (scFv) library against TNFα was constructed using a recombinant phage antibody system (RPAS). The cloned scFv gene was introduced into the phage display vector pCANTAB 5E and expressed in Escherichia coli (E. coli) with a yield of up to 0.15 mg/l of total protein. With the attempt to improve the expression level of TNF-scFv, a strategy was established for subcloning the scFv gene from pCANTAB 5E into the plasmid pBV220. Under the control of a highly efficient tandem P(R)P(L) promoter system, scFv production was increased to 30% of total protein as inclusion bodies. After extraction from the cell pellet by sonication, the inclusion bodies were solubilized and denatured in the presence of 8M urea. Purification of denatured scFv was performed using nickel column chromatography followed by renaturation. The purity and activity of the refolded scFv were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), Western blotting and by an enzyme-linked immunoabsorbent assay (ELISA). The results reveal that the overall yield of bioactive TNF-scFv from E. coli flask cultures was more than 45 mg/l culture medium and 15 mg/g wet weight cells. The renatured scFv exhibited binding activity similarly to soluble scFv. In conclusion we developed a method to over-express TNF-scFv, which have biological function after purification and renaturation. Copyright © 2010 Elsevier Inc. All rights reserved.

Human scFv antibody fragments specific for hepatocellular carcinoma selected from a phage display library.

PubMed

Yu, Bing; Ni, Ming; Li, Wen-Han; Lei, Ping; Xing, Wei; Xiao, Dai-Wen; Huang, Yu; Tang, Zhen-Jie; Zhu, Hui-Fen; Shen, Guan-Xin

2005-07-14

To identify the scFv antibody fragments specific for hepatocellular carcinoma by biopanning from a large human naive scFv phage display library. A large human naive scFv phage library was used to search for the specific targets by biopanning with the hepatocellular carcinoma cell line HepG2 for the positive-selecting and the normal liver cell line L02 for the counter-selecting. After three rounds of biopanning, individual scFv phages binding selectively to HepG2 cells were picked out. PCR was carried out for identification of the clones containing scFv gene sequence. The specific scFv phages were selected by ELISA and flow cytometry. DNA sequences of positive clones were analyzed by using Applied Biosystem Automated DNA sequencers 3 730. The expression proteins of the specific scFv antibody fragments in E.coli HB2151 were purified by the affinity chromatography and detected by SDS-PAGE, Western blot and ELISA. The biological effect of the soluble antibody fragments on the HepG2 cells was investigated by observing the cell proliferation. Two different positive clones were obtained and the functional variable sequences were identified. Their DNA sequences of the scFv antibody fragments were submitted to GenBank (accession nos: AY686498 and AY686499). The soluble scFv antibody fragments were successfully expressed in E.coli HB2151. The relative molecular mass of the expression products was about 36 ku, according to its predicted M(r) value. The two soluble scFv antibody fragments also had specific binding activity and obvious growth inhibition properties to HepG2 cells. The phage library biopanning permits identification of specific antibody fragments for hepatocellular carcinoma and affords experiment evidence for its immunotherapy study.

Construction, expression and in vitro biological behaviors of Ig scFv fragment in patients with chronic B cell leukemia.

PubMed

Zhu, Lijuan; Liao, Wenjun; Zhu, Huifen; Lei, Ping; Wang, Zhihua; Shao, Jingfang; Zhang, Yue; Shen, Guanxin

2006-01-01

The expression vector of SmIg scFv fragment was constructed in patient with B cell chronic lymphocyte leukemia (B-CLL) and expressed in E. coli to obtain scFv fragment, and the effect of the protein on the proliferation of stimulated peripheral blood mononuclear cells (PBMC) was investigated in vitro. Two pairs of primers were designed, and variable region genes of light chain and heavy chain were amplified by PCR respectively from the pGEM-T vectors previously constructed in our laboratory which containing light chain gene or Fd fragment of heavy chain gene. The PCR product was digested, purified and inserted into pHEN2 vector to construct the soluble expression vector pHEN2-scFv. After the induction by IPTG, the scFv protein was identified by SDS-PAGE electrophoresis and purified by Ni-NTA-Chromatography. MTT was used to determine the effect of purified protein on the proliferation of stimulated PBMC in vitro. Plasmid PCR and restriction enzyme digestion of pHEN2-scFv revealed the pHEN2-scFv vector was constructed successfully. Id-scFv protein was expressed in positive clone after induced by IPTG. SDS-PAGE analysis showed that the relative molecular weight of fusion protein was about 30 kD (1 kD= 0.9921 ku), which was consistent with the theoretically predicted value. Proliferation of PBMC could be induced by purified Id-scFv. It was suggested that the expression vector of SmIg scFv fragment was constructed successfully, and scFv protein was expressed and secreted from E. coli, which could induce proliferation of PBMC. This may lay an experimental foundation for further research of Id-HSP complex vaccine for B-CLL.

Fruits and vegetables as a healthier snack throughout the day among families with older children: findings from a survey of parent-child dyads.

PubMed

Smith, Teresa M; Pinard, Courtney A; Byker Shanks, Carmen; Wethington, Holly; Blanck, Heidi M; Yaroch, Amy L

2015-04-01

Most U.S. youth fail to eat the recommended amount of fruits and vegetables (FV) however many consume too many calories as added sugars and solid fats, often as snacks. The aim of this study was to assess factors associated with serving FV as snacks and with meals using parent-child dyads. A cross-sectional sample of U.S. children aged 9 to 18, and their caregiver/parent (n=1522) were part of a Consumer Panel of households for the 2008 YouthStyles mail survey. Chi-square test of independence and multivariable logistic regression were used to assess associations between serving patterns of FV as snacks with variations in serving patterns, and covariates including dietary habits. Most parents (72%) reported serving FV at meals and as snacks. Fruit was most frequently served as a snack during the day (52%) and vegetables were most frequently served as a snack during the day (22%) but rarely in the morning. Significant differences in child FV intake existed among FV as a snack serving patterns by parents. Compared to children whose parents served FV only at meals, children whose parents reported serving FV as snacks in addition to meals were significantly more likely to have consumed FV the day before (using a previous day screener), P<0.05. Contributing to the growing collection of literature describing parent-child dyad dietary behaviors, these findings suggest promoting FV access and intake throughout the day, not only at meals, by including serving as snacks, may increase FV intake among older children and adolescents. Copyright © 2015 Elsevier Ltd. All rights reserved.

Exploring perceptions and beliefs about the cost of fruit and vegetables and whether they are barriers to higher consumption.

PubMed

Chapman, Kathryn; Goldsbury, David; Watson, Wendy; Havill, Michelle; Wellard, Lyndal; Hughes, Clare; Bauman, Adrian; Allman-Farinelli, Margaret

2017-06-01

Fruit and vegetable (F&V) consumption is below recommendations, and cost may be a barrier to meeting recommendations. Limited evidence exists on individual perceptions about the cost, actual spending and consumption of F&V. This study investigated perceptions and beliefs about cost of F&V and whether this is a barrier to higher consumption. An online survey of Australian adults (n = 2474) measured F&V consumption; expenditure on F&V and food; and perceived barriers to consumption. Multivariable logistic regression examined associations between participants' responses about cost of F&V and demographic factors, and with actual consumption and expenditure on F&V. Cost was identified as a barrier for 29% of people not meeting recommended fruit servings and for 14% of people not meeting recommendations for vegetables. Cost was a more common barrier for those on lower incomes (fruit aOR 1.89; 95% CI 1.20-2.98 and vegetables aOR 2.94; 95% CI 1.97-4.39) and less common for older participants (fruit aOR 0.33; 95% CI 0.17-0.62 and vegetables aOR 0.31; 95% CI 0.18-0.52). There was no association between the perceived barriers and actual F&V spending. Twenty percent of participants said F&V were not affordable; 39% said cost made it difficult to buy F&V, and for 23% the cost of F&V meant they bought less than desired. A minority reported F&V were not affordable where they shopped and that cost was a barrier to higher consumption. However, it is apparent that young adults and those on low incomes eat less than they would like because of cost. Strategies that remove financial impediments to consumption are indicated for these population sub-groups. Copyright © 2017 Elsevier Ltd. All rights reserved.

Development of a microplate coagulation assay for Factor V in human plasma.

PubMed

Tilley, Derek; Levit, Irina; Samis, John A

2011-06-28

Factor V (FV) in its activated form, FVa, is a critical regulator of thrombin generation during fibrin clot formation. There is a need of a simple, fast, and inexpensive microplate-based coagulation assay to measure the functional activity of FV in human plasma. The objective of this study was to develop a microplate-based assay that measures FV coagulation activity during clot formation in human plasma, which is currently not available. The FV assay requires a kinetic microplate reader to measure the change in absorbance at 405nm during fibrin formation in human plasma. The FV assay accurately measures the time, initial rate, and extent of fibrin clot formation in human plasma. The FV microplate assay is simple, fast, economical, sensitive to approx 24-80pM, and multiple samples may be analyzed simultaneously. All the required materials are commercially available. Standard curves of time or initial rate of fibrin clot formation vs FV activity in the 1-stage assay (Without activation by thrombin) may be used to measure FV activity in samples of human plasma. The assay was used to demonstrate that in nine patients with disseminated intravascular coagulation (DIC), the FV 1-stage, 2-stage (With activation by thrombin), and total (2-stage activity - 1-stage activity) activities were decreased, on average, by approximately 54%, 44%, and 42%, respectively, from prolonged clot times when compared to normal pooled human reference plasma (NHP). The results indicate that the FV in the DIC patient plasmas supported both a delayed and slower rate of fibrin clot formation compared with NHP; however, the extent of fibrin clot formation in the DIC patients remained largely unchanged from that observed with NHP. The FV microplate assay may be easily adapted to measure the activity of any coagulation factor using the appropriate factor-deficient plasma and clot initiating reagent. The microplate assay will find use in both research and clinical laboratories to provide measurement of the functional coagulation activity of FV in human plasma.

In vitro caries lesion rehardening and enamel fluoride uptake from fluoride varnishes as a function of application mode.

PubMed

Lippert, Frank; Hara, Anderson T; Martinez-Mier, Esperanza Angeles; Zero, Domenick T

2013-04-01

To study the laboratory predicted anticaries efficacy of five commercially available fluoride varnishes (FV) by determining their ability to reharden and to deliver fluoride to an early caries lesion when applied directly or in close vicinity to the lesion (halo effect). Early caries lesions were created in 80 polished bovine enamel specimens. Specimens were allocated to five FV groups (n = 16) based on Knoop surface microhardness (KHN) after lesion creation. All tested FV claimed to contain 5% sodium fluoride and were: CavityShield, Enamel Pro, MI Varnish, Prevident and Vanish. FV were applied (10 +/- 2 mg per lesion) to eight specimens per FV group (direct application); the remaining eight specimens received no FV but were later exposed to fluoride released from specimens which received a FV treatment (indirect application). Specimens were paired again and placed into containers (one per FV). Artificial saliva was added and containers placed into an incubator (27 hours at 37 degrees C). Subsequently, FV was carefully removed using chloroform. Specimens were exposed to fresh artificial saliva again (67 hours at 37 degrees C). KHN was measured and differences to baseline values calculated. Enamel fluoride uptake (EFU) was determined using the acid etch technique. Data were analyzed using two-way ANOVA. The two-way ANOVA highlighted significant interactions between FV vs. application mode, for both deltaKHN and EFU (P < 0.001). All FV were able to reharden and deliver fluoride to caries lesions, but to different degrees. Furthermore, considerable differences were found for both variables between FV when applied either directly or in close vicinity to the lesion: MI Varnish and Enamel Pro exhibited greater fluoride efficacy when applied in vicinity rather than directly to the lesion, whereas CavityShield and Vanish did not differ. Prevident exhibited a higher EFU when applied directly, but little difference in rehardening.

The combination of Cassia obtusifolia L. and Foeniculum vulgare M. exhibits a laxative effect on loperamide-induced constipation of rats

PubMed Central

Jang, Seung Hee

2018-01-01

Chronic constipation is a functional gastrointestinal disease that is detrimental to the quality of patient life. Cassia obtusifolia L. (CO) and Foeniculum vulgare M. (FV) are commonly used as medicinal foods in many countries. We aimed to examine the laxative effect and their underlying mechanism of CO and FV mixture on loperamide (lop)-induced constipated rats. To determine the laxative effects of these compounds, Sprague-Dawley rats were divided into six groups: the control, lop-induced constipated (2mg/kg), and three doses (100, 300, and 500mg/kg) of CO and FV mixture-, and Bisacodyl (bis, 3.3mg/kg)-treated groups. The mixture of CO and FV and bis were orally administered once a day for 4 weeks. For induction of constipation, the lop were treated with a dose of 2 mg/kg twice a day on the 3rd week after treatments of CO and FV extracts and bis. The results were revealed that the CO and FV mixture has the laxative effects more than those in CO and FV-alone treatments on constipated rats by determining the stool parameters, including stool number and weight. Indeed, stool parameters, such as, stool number, weight, and water contents and colonic peristalsis from the intestinal transit length and ratio were dramatically improved by CO and FV mixture treatment. Histological study also revealed that CO and FV mixture enhanced the thicknesses of mucosa and muscular layers of the colon in constipated rats. For their underlying mechanism, the mRNAs and proteins expression of muscarinic acetylcholine receptors (mAchR) M2 and M3 and their downstream signaling were preserved by CO and FV mixture treatment in constipated rats. Therefore, this study suggests that treatment with CO and FV mixture has beneficial effects against constipation. We further suggest that CO and FV mixture may be utilized as an alternative therapeutic strategy for constipation. PMID:29621360

Complementarity determining regions and frameworks contribute to the disulfide bond independent folding of intrinsically stable scFv

PubMed Central

Gąciarz, Anna

2017-01-01

CyDisCo is a system facilitating disulfide bond formation in recombinant proteins in the cytoplasm of Escherichia coli. Previously we screened for soluble expression of single chain antibody fragments (scFv) in the cytoplasm of E. coli in the presence and absence of CyDisCo, with >90% being solubly expressed. Two scFv, those derived from natalizumab and trastuzumab, were solubly produced in high amounts even in the absence of folding catalysts i.e. disulfide bond formation is not critical for their folding. Here we investigate the contribution of the framework and the complementarity determining regions (CDRs) of scFv to the disulfide-independence of folding. We swapped CDRs between four scFv that have different properties, including two scFv that can efficiently fold independently from disulfide bonds and two more disulfide-dependent scFv. To confirm disulfide-independence we generated cysteine to alanine mutants of the disulfide-independent scFv. All of the scFv were tested for soluble expression in the cytoplasm of E. coli in the presence and absence of the oxidative folding catalysts Erv1p and PDI. Eight of the hybrid scFv were solubly produced in the presence of CyDisCo, while seven were solubly produced in the absence of CyDisCo, though the yields were often much lower when CyDisCo was absent. Soluble expression was also observed for scFv natalizumab and trastuzumab containing no cysteines. We compared yields, thermal stability and secondary structure of solubly produced scFv and undertook binding studies by western blotting, dot blotting or surface plasmon resonance of those produced in good yields. Our results indicate that both the CDRs and the framework contribute to the disulfide-dependence of soluble production of scFv, with the CDRs having the largest effect. In addition, there was no correlation between thermal stability and disulfide-dependence of folding and only a weak correlation between the yield of protein and the thermal stability of the protein. PMID:29253024

Complementarity determining regions and frameworks contribute to the disulfide bond independent folding of intrinsically stable scFv.

PubMed

Gąciarz, Anna; Ruddock, Lloyd W

2017-01-01

CyDisCo is a system facilitating disulfide bond formation in recombinant proteins in the cytoplasm of Escherichia coli. Previously we screened for soluble expression of single chain antibody fragments (scFv) in the cytoplasm of E. coli in the presence and absence of CyDisCo, with >90% being solubly expressed. Two scFv, those derived from natalizumab and trastuzumab, were solubly produced in high amounts even in the absence of folding catalysts i.e. disulfide bond formation is not critical for their folding. Here we investigate the contribution of the framework and the complementarity determining regions (CDRs) of scFv to the disulfide-independence of folding. We swapped CDRs between four scFv that have different properties, including two scFv that can efficiently fold independently from disulfide bonds and two more disulfide-dependent scFv. To confirm disulfide-independence we generated cysteine to alanine mutants of the disulfide-independent scFv. All of the scFv were tested for soluble expression in the cytoplasm of E. coli in the presence and absence of the oxidative folding catalysts Erv1p and PDI. Eight of the hybrid scFv were solubly produced in the presence of CyDisCo, while seven were solubly produced in the absence of CyDisCo, though the yields were often much lower when CyDisCo was absent. Soluble expression was also observed for scFv natalizumab and trastuzumab containing no cysteines. We compared yields, thermal stability and secondary structure of solubly produced scFv and undertook binding studies by western blotting, dot blotting or surface plasmon resonance of those produced in good yields. Our results indicate that both the CDRs and the framework contribute to the disulfide-dependence of soluble production of scFv, with the CDRs having the largest effect. In addition, there was no correlation between thermal stability and disulfide-dependence of folding and only a weak correlation between the yield of protein and the thermal stability of the protein.

Healthier side dishes at restaurants: an analysis of children's perspectives, menu content, and energy impacts.

PubMed

Anzman-Frasca, Stephanie; Dawes, Franciel; Sliwa, Sarah; Dolan, Peter R; Nelson, Miriam E; Washburn, Kyle; Economos, Christina D

2014-07-04

Children consume restaurant-prepared foods at high rates, suggesting that interventions and policies targeting consumption of these foods have the potential to improve diet quality and attenuate excess energy intake. One approach to encouraging healthier dietary intake in restaurants is to offer fruits and vegetables (FV) as side dishes, as opposed to traditional, energy-dense accompaniments like French fries. The aims of the current study were to examine: children's views about healthier side dishes at restaurants; current side dish offerings on children's menus at leading restaurants; and potential energy reductions when substituting FV side dishes in place of French fries. To investigate children's attitudes, a survey was administered to a nationally representative sample of U.S. 8- to 18-year-olds (n = 1178). To examine current side dish offerings, children's menus from leading quick service (QSR; n = 10) and full service restaurant chains (FSR; n = 10) were analyzed. Energy reductions that could result from substituting commonly-offered FV side dishes for French fries were estimated using nutrition information corresponding to the children's menu items. Two-thirds of children reported that they would not feel negatively about receiving FV sides instead of French fries with kids' meals. Liking/taste was the most common reason that children gave to explain their attitudes about FV side dishes. Nearly all restaurants offered at least 1 FV side dish option, but at most restaurants (60% of QSR; 70% of FSR), FV sides were never served by default. Substituting FV side dishes for French fries yielded an average estimated energy reduction of at least 170 calories. Results highlight some healthy trends in the restaurant context, including the majority of children reporting non-negative attitudes about FV side dishes and the consistent availability of FV side dish options at leading QSR and FSR. Yet the minority of restaurants offer these FV sides by default. Promoting creative, appealing FV side dishes can result in healthier, less energy-dense meals for children. Substituting or displacing energy-dense default side dishes with such FV dishes show promise as part of continued, comprehensive efforts to increase the healthfulness of meals consumed by children in restaurant settings.

On carrots and curiosity: eating fruit and vegetables is associated with greater flourishing in daily life.

PubMed

Conner, Tamlin S; Brookie, Kate L; Richardson, Aimee C; Polak, Maria A

2015-05-01

Our aim was to determine whether eating fruit and vegetables (FV) is associated with other markers of well-being beyond happiness and life satisfaction. Towards this aim, we tested whether FV consumption is associated with greater eudaemonic well-being - a state of flourishing characterized by feelings of engagement, meaning, and purpose in life. We also tested associations with two eudaemonic behaviours - curiosity and creativity. Daily diary study across 13 days (micro-longitudinal, correlational design). A sample of 405 young adults (67% women; mean age 19.9 [SD 1.6] years) completed an Internet daily diary for 13 consecutive days. Each day, participants reported on their consumption of fruit, vegetables, sweets, and chips, as well as their eudaemonic well-being, curiosity, creativity, positive affect (PA), and negative affect. Between-person associations were analysed on aggregated data. Within-person associations were analysed using multilevel models controlling for weekday and weekend patterns. Fruit and vegetables consumption predicted greater eudaemonic well-being, curiosity, and creativity at the between- and within-person levels. Young adults who ate more FV reported higher average eudaemonic well-being, more intense feelings of curiosity, and greater creativity compared with young adults who ate less FV. On days when young adults ate more FV, they reported greater eudaemonic well-being, curiosity, and creativity compared with days when they ate less FV. FV consumption also predicted higher PA, which mostly did not account for the associations between FV and the other well-being variables. Few unhealthy foods (sweets, chips) were related to well-being except that consumption of sweets was associated with greater curiosity and PA at the within-person level. Lagged data analyses showed no carry-over effects of FV consumption onto next-day well-being (or vice versa). Although these patterns are strictly correlational, this study provides the first evidence that FV consumption may be related to a broader range of well-being states that signal human flourishing in early adulthood. Statement of contribution What is already known on this subject? There is growing evidence that a diet rich in fruits and vegetables (FV) is related to greater happiness, life satisfaction, and positive affect. These associations are not entirely explained by demographic or health variables including socio-economic status, exercise, smoking, and body mass index (BMI). Recent experimental and daily diary research suggests that FV consumption may be a causal factor in promoting states of positive well-being. Research has examined the links between FV consumption and hedonic well-being - whether people feel good (vs. bad) and satisfied-but has not addressed links between FV consumption and eudaemonic well-being- whether people feel engaged and experience their lives as meaningful and purposeful. What does this study add? It provides the first evidence that eating FV is related to greater eudaemonic well-being in a naturalistic setting. Eating FV was also related to greater self-reported curiosity and creativity. FV consumption may underlie a broad range of experiences that signal flourishing. Future randomised controlled trials of FV should include measures of eudaemonic well-being as outcome variables. © 2014 The British Psychological Society.

Does Nutrition Education with Fruit and Vegetable Supplementation Increase Fruit and Vegetable Intake and Improve Anthropometrics of Overweight or Obese People of Varying Socioeconomic Status?

PubMed

Honrath, Kerrie; Wagner, Meredith G; Rhee, Yeong

2018-01-01

Fruit and vegetable (F/V) intake is inadequate and obesity is more prevalent among adults of lower socioeconomic status (SES) in the United States. The effect of nutrition education and F/V supplementation on F/V intake and anthropometrics of overweight or obese adults of varying SES was determined. F/V intake was not different between the nutrition education and F/V supplementation groups. Individuals with a graduate degree had significant improvements in fruit intake. Few of the improvements in anthropometrics seen were significant. Future research should focus on specific barriers to F/V intake and include information on total energy intake and expenditure.

Numerical investigation of finite-volume effects for the HVP

NASA Astrophysics Data System (ADS)

Boyle, Peter; Gülpers, Vera; Harrison, James; Jüttner, Andreas; Portelli, Antonin; Sachrajda, Christopher

2018-03-01

It is important to correct for finite-volume (FV) effects in the presence of QED, since these effects are typically large due to the long range of the electromagnetic interaction. We recently made the first lattice calculation of electromagnetic corrections to the hadronic vacuum polarisation (HVP). For the HVP, an analytical derivation of FV corrections involves a two-loop calculation which has not yet been carried out. We instead calculate the universal FV corrections numerically, using lattice scalar QED as an effective theory. We show that this method gives agreement with known analytical results for scalar mass FV effects, before applying it to calculate FV corrections for the HVP. This method for numerical calculation of FV effects is also widely applicable to quantities beyond the HVP.

Mediation of parental educational level on fruit and vegetable intake among schoolchildren in ten European countries.

PubMed

Lehto, Elviira; Ray, Carola; Te Velde, Saskia; Petrova, Stefka; Duleva, Vesselka; Krawinkel, Michael; Behrendt, Isabel; Papadaki, Angeliki; Kristjansdottir, Asa; Thorsdottir, Inga; Yngve, Agneta; Lien, Nanna; Lynch, Christel; Ehrenblad, Bettina; Vaz de Almeida, Maria Daniel; Ribic, Cirila Hlastan; Simčic, Irena; Roos, Eva

2015-01-01

To examine which factors act as mediators between parental educational level and children's fruit and vegetable (F&V) intake in ten European countries. Cross-sectional data were collected in ten European countries participating in the PRO GREENS project (2009). Schoolchildren completed a validated FFQ about their daily F&V intake and filled in a questionnaire about availability of F&V at home, parental facilitation of F&V intake, knowledge of recommendations about F&V intake, self-efficacy to eat F&V and liking for F&V. Parental educational level was determined from a questionnaire given to parents. The associations were examined with multilevel mediation analyses. Schools in Bulgaria, Finland, Germany, Greece, Iceland, the Netherlands, Norway, Portugal, Slovenia and Sweden. Eleven-year-old children (n 8159, response rate 72%) and their parents. In five of the ten countries, children with higher educated parents were more likely to report eating fruits daily. This association was mainly mediated by knowledge but self-efficacy, liking, availability and facilitation also acted as mediators in some countries. Parents' education was positively associated with their children's daily vegetable intake in seven countries, with knowledge and availability being the strongest mediators and self-efficacy and liking acting as mediators to some degree. Parental educational level correlated positively with children's daily F&V intake in most countries and the pattern of mediation varied among the participating countries. Future intervention studies that endeavour to decrease the educational-level differences in F&V intake should take into account country-specific features in the relevant determinants of F&V intake.

Identification, isolation, and expression analysis of heat shock transcription factors in the diploid woodland strawberry Fragaria vesca

PubMed Central

Hu, Yang; Han, Yong-Tao; Wei, Wei; Li, Ya-Juan; Zhang, Kai; Gao, Yu-Rong; Zhao, Feng-Li; Feng, Jia-Yue

2015-01-01

Heat shock transcription factors (Hsfs) are known to play dominant roles in plant responses to heat, as well as other abiotic or biotic stress stimuli. While the strawberry is an economically important fruit plant, little is known about the Hsf family in the strawberry. To explore the functions of strawberry Hsfs in abiotic and biotic stress responses, this study identified 17 Hsf genes (FvHsfs) in a wild diploid woodland strawberry (Fragaria vesca, 2n = 2x = 14) and isolated 14 of these genes. Phylogenetic analysis divided the strawberry FvHsfs genes into three main groups. The evolutionary and structural analyses revealed that the FvHsf family is conserved. The promoter sequences of the FvHsf genes contain upstream regulatory elements corresponding to different stress stimuli. In addition, 14 FvHsf-GFP fusion proteins showed differential subcellular localization in Arabidopsis mesophyll protoplasts. Furthermore, we examined the expression of the 17 FvHsf genes in wild diploid woodland strawberries under various conditions, including abiotic stresses (heat, cold, drought, and salt), biotic stress (powdery mildew infection), and hormone treatments (abscisic acid, ethephon, methyl jasmonate, and salicylic acid). Fifteen of the seventeen FvHsf genes exhibited distinct changes on the transcriptional level during heat treatment. Of these 15 FvHsfs, 8 FvHsfs also exhibited distinct responses to other stimuli on the transcriptional level, indicating versatile roles in the response to abiotic and biotic stresses. Taken together, the present work may provide the basis for further studies to dissect FvHsf function in response to stress stimuli. PMID:26442049

The value of facial attractiveness for encouraging fruit and vegetable consumption: analyses from a randomized controlled trial.

PubMed

Appleton, Katherine M; McGrath, Alanna J; McKinley, Michelle C; Draffin, Claire R; Hamill, Lesley L; Young, Ian S; Woodside, Jayne V

2018-03-01

An effect of increased fruit and vegetable (FV) consumption on facial attractiveness has been proposed and recommended as a strategy to promote FV intakes, but no studies to date demonstrate a causal link between FV consumption and perceived attractiveness. This study investigated perceptions of attractiveness before and after the supervised consumption of 2, 5 or 8 FV portions/day for 4 weeks in 30 low FV consumers. Potential mechanisms for change via skin colour and perceived skin healthiness were also investigated. Faces were photographed at the start and end of the 4 week intervention in controlled conditions. Seventy-three independent individuals subsequently rated all 60 photographs in a randomized order, for facial attractiveness, facial skin yellowness, redness, healthiness, clarity, and symmetry. Using clustered multiple regression, FV consumption over the previous 4 weeks had no direct effect on attractiveness, but, for female faces, some evidence was found for an indirect impact, via linear and non-linear changes in skin yellowness. Effect sizes, however, were small. No association between FV consumption and skin healthiness was found, but skin healthiness was associated with facial attractiveness. Controlled and objectively measured increases in FV consumption for 4 weeks resulted indirectly in increased attractiveness in females via increases in skin yellowness, but effects are small and gradually taper as FV consumption increases. Based on the effect sizes from this study, we are hesitant to recommend the use of facial attractiveness to encourage increased FV consumption. Clinical trial Registration Number NCT01591057 ( www.clinicaltrials.gov ). Registered: 27th April, 2012.

Changes in fruit and vegetable consumption of third-grade students in body quest: food of the warrior, a 17-class childhood obesity prevention program.

PubMed

Struempler, Barbara J; Parmer, Sondra M; Mastropietro, Lisa M; Arsiwalla, Dilbur; Bubb, Robert R

2014-01-01

To increase fruit and vegetable (FV) consumption of youth in Body Quest: Food of the Warrior (BQ), a childhood obesity prevention program. Quasi-experimental. Supplemental Nutrition Assistance Program-Education eligible schools (n = 60). Third-grade students (n = 2,477). Treatment groups (n = 1,674) self-reported foods consumed through the School Lunch Program for 17 weekly assessments; they participated in BQ curriculum, iPad app education, and weekly FV tastings. Control groups (n = 803) completed only pre- and post-assessments. Weekly FV consumed through School Lunch Program. ANCOVA and growth modeling. From before to after the program, the treatment group demonstrated significant, moderate increases in fruit (P < .01) and vegetable (P < .001) consumptions, increasing from 7 to 8 weekly FV servings. After the program, the treatment group consumed significantly (P < .001) more FV than the control group. Fruit and vegetable consumption increased to class 10 and then stabilized. From before to after the program, all FV predictors were significantly higher and included gender (vegetables), race (FV), and free/reduced lunch (fruit). Nutrition programs can increase FV intake. Even moderate increases in FV intake can be an initial step for the prevention of chronic disease. Copyright © 2014 Society for Nutrition Education and Behavior. Published by Elsevier Inc. All rights reserved.

A Farmers’ Market at a Federally Qualified Health Center Improves Fruit and Vegetable Intake among Low-income Diabetics

PubMed Central

Freedman, Darcy A.; Choi, Seul Ki; Hurley, Thomas; Anadu, Edith; Hebert, James R.

2013-01-01

Objective A 22-week federally qualified health center (FQHC)-based farmers’ market (FM) and personal financial incentive intervention designed to improve access to and consumption of fruits and vegetables (FV) among low-income diabetics in rural South Carolina was evaluated. Methods A mixed methods, one-group, repeated-measures design was used. Data were collected in 2011 before (May/June), during (August), and after (November) the intervention with 41 diabetes patients from the FQHC. FV consumption was assessed using a validated National Cancer Institute FV screener modified to include FV sold at the FM. Sales receipts were recorded for all FM transactions. A mixed-model, repeated measures analysis of variance was used to assess intervention effects on FV consumption. Predictors of changes in FV consumption were examined using logistic regression. Results A marginally significant (p=0.07) average increase of 1.6 servings of total FV consumption per day occurred. The odds of achieving significant improvements in FV consumption increased for diabetics using financial incentives for payment at the FM (OR: 38.8, 95% CI: 3.4–449.6) and for those frequenting the FM more often (OR: 2.1, 95% CI: 1.1–4.0). Conclusions Results reveal a dose-response relationship between the intervention and FV improvements and emphasize the importance of addressing economic barriers to food access. PMID:23384473

[Relationship between factor v Leiden mutation and Chinese Budd-Chiari syndrome and its clinical significance].

PubMed

Feng, B; Xu, K; Jiang, H

2000-05-01

To investigate the relationship between factor v Leiden (FvL) mutation and Chinese sporadic Budd-Chiari syndrome (BCS), familial BCS, and to explore the significance of FvL mutation in the etiology of BCS. Twenty-five patients with sporadic BCS, 6 patients with familial BCS (from A and B families), 39 both A and B family members, and 31 healthy persons were detected for FvL mutation with PCR. Meantime, two family members were explored for the related etiology of BCS. Factor V Leiden mutation was detected in 4 of 6 patients with familial BCS and in 2 family members. AIII(7,11,15) and BII(10), AII(2) and BIII(5) were found FvL mutation, and mutation was heterzygous. FvL mutation in the two degrees was compatible with Mendel hereditery law. The frequency of FvL mutation in 31 BCS and 31 healthy persons showed no statistical significance: but the frequency of FvL mutation between the familial BCS and healthy persons showed statistical significance. Ten persons in A family had varicose vein of the low extremeties, which was compatible with autosomal dominant inheritance. FvL mutation is related to Chinese familial BCS, but is not related to Chinese sporadic BCS. FvL mutation may be a underlying pathogenicity of familial BCS. Varicose vein of the low extremeties may be one of the pathogenicity of familial BCS.

Adolescent Overweight and Obesity: Links to Socioeconomic Status and Fruit and Vegetable Intakes

PubMed Central

You, Jihyun; Choo, Jina

2016-01-01

Whether adolescent overweight/obesity is linked to socioeconomic status (SES) and fruit and vegetable (F/V) intakes has not been confirmed. We aimed to determine whether there is an association between SES and adolescent overweight/obesity and to test the mediating effect of F/V intakes. This cross-sectional study included the data of 63,111 adolescents extracted from the 2013 Korea Youth Risk Behavior Web-based Survey. Overweight/obesity was defined as a body mass index ≥ 85th percentile, while F/V intakes were categorized as high (recommended levels: ≥1 fruit serving and ≥3 vegetable servings per day) versus low. Among girls, low SES (beta = 0.50, p < 0.001) and F/V intakes (beta = −0.17, p = 0.038) were both significantly associated with overweight/obesity; the former association was significantly mediated by F/V intakes (Sobel test: z = 2.00, p = 0.046). Among boys, neither SES nor F/V intakes was significantly associated with overweight/obesity. Adolescent overweight/obesity was significantly linked to low SES and F/V intakes among girls only; low SES indirectly increased the risk of overweight/obesity via low F/V intakes. Therefore, promoting F/V intakes for socially disadvantaged girls should be prioritized as a population-based strategy for preventing adolescent overweight/obesity in South Korea. PMID:27005654

Fruit and Vegetable Attitudes, Norms, and Intake in Low-Income Youth

ERIC Educational Resources Information Center

Di Noia, Jennifer; Cullen, Karen Weber

2015-01-01

Fruit and vegetable (FV) attitudes and norms have been shown to influence intake in youth; yet research with low-income youth and studies supplementing self-report with objective measures of intake are lacking. Cross-sectional survey data on self-rated FV intake, FV attitudes, and FV norms were collected in a sample of 116 youth attending a…

Examination of the Relationship between In-Store Environmental Factors and Fruit and Vegetable Purchasing among Hispanics.

PubMed

Sanchez-Flack, Jennifer; Pickrel, Julie L; Belch, George; Lin, Shih-Fan; Anderson, Cheryl A M; Martinez, Maria Elena; Arredondo, Elva M; Ayala, Guadalupe X

2017-10-27

Retail food environments have received attention for their influence on dietary behaviors and for their nutrition intervention potential. To improve diet-related behaviors, such as fruit and vegetable (FV) purchasing, it is important to examine its relationship with in-store environmental characteristics. This study used baseline data from the " El Valor de Nuestra Salud " study to examine how in-store environmental characteristics, such as product availability, placement and promotion, were associated with FV purchasing among Hispanic customers in San Diego County. Mixed linear regression models indicated that greater availability of fresh FVs was associated with a $0.36 increase in FV purchasing ( p = 0.01). Placement variables, specifically each additional square foot of display space dedicated to FVs ( p = 0.01) and each additional fresh FV display ( p = 0.01), were associated with a $0.02 increase and $0.29 decrease, respectively, in FV purchasing. Introducing FV promotions in the final model was not related to FV purchasing. Exploratory analyses indicated that men reported spending $3.69 fewer dollars on FVs compared to women, controlling for covariates ( p = 0.02). These results can help inform interventions targeting in-store environmental characteristics to encourage FV purchasing among Hispanics.

Production of in vivo biotinylated scFv specific to almond (Prunus dulcis) proteins by recombinant Pichia pastoris.

PubMed

de la Cruz, Silvia; Alcocer, Marcos; Madrid, Raquel; García, Aina; Martín, Rosario; González, Isabel; García, Teresa

2016-06-10

The methylotropic yeast Pichia pastoris has demonstrated its suitability for large-scale production of recombinant proteins. As an eukaryotic organism P. pastoris presents a series of advantages at expression and processing of heterologous proteins when compared with Escherichia coli. In this work, P. pastoris has been used to express a scFv from a human synthetic library previously shown to bind almond proteins. In order to facilitate purification and post processing manipulations, the scFv was engineered with a C-terminal tag and biotinylated in vivo. After purification, biotinylated scFv were bound to avidin conjugated with HRP producing a multimeric scFv. The multimeric scFv showed to maintain their ability to recognize almond protein when assayed in ELISA, reaching a LOD of 470mgkg(-1). This study describes an easy method to produce large quantities of in vivo biotinylated scFv in P. pastoris. By substituting the enzyme or fluorochromes linked to avidin, it will be possible to generate a diverse number of multimeric scFv as probes to suit different analytical platforms in the detection of almond in food products. Copyright © 2016 Elsevier B.V. All rights reserved.

Anti-CTGF single-chain variable fragment dimers inhibit human airway smooth muscle (ASM) cell proliferation by down-regulating p-Akt and p-mTOR levels.

PubMed

Gao, Wei; Cai, Liting; Xu, Xudong; Fan, Juxiang; Xue, Xiulei; Yan, Xuejiao; Qu, Qinrong; Wang, Xihua; Zhang, Chen; Wu, Guoqiu

2014-01-01

Connective tissue growth factor (CTGF) contributes to airway smooth muscle (ASM) cell hyperplasia in asthma. Humanized single-chain variable fragment antibody (scFv) was well characterized as a CTGF antagonist in the differentiation of fibroblast into myofibroblast and pulmonary fibrosis in our previous studies. To further improve the bioactivity of scFv, we constructed a plasmid to express scFv-linker-matrilin-6×His fusion proteins that could self-assemble into the scFv dimers by disulfide bonds in matrilin under non-reducing conditions. An immunoreactivity assay demonstrated that the scFv dimer could highly bind to CTGF in a concentration-dependent manner. The MTT and EdU assay results revealed that CTGF (≥10 ng/mL) promoted the proliferation of ASM cells, and this effect was inhibited when the cells were treated with anti-CTGF scFv dimer. The western blot analysis results showed that increased phosphorylation of Akt and mTOR induced by CTGF could be suppressed by this scFv dimer. Based on these findings, anti-CTGF scFv dimer may be a potential agent for the prevention of airway remodeling in asthma.

Anti-CTGF Single-Chain Variable Fragment Dimers Inhibit Human Airway Smooth Muscle (ASM) Cell Proliferation by Down-Regulating p-Akt and p-mTOR Levels

PubMed Central

Xu, Xudong; Fan, Juxiang; Xue, Xiulei; Yan, Xuejiao; Qu, Qinrong; Wang, Xihua; Zhang, Chen; Wu, Guoqiu

2014-01-01

Connective tissue growth factor (CTGF) contributes to airway smooth muscle (ASM) cell hyperplasia in asthma. Humanized single-chain variable fragment antibody (scFv) was well characterized as a CTGF antagonist in the differentiation of fibroblast into myofibroblast and pulmonary fibrosis in our previous studies. To further improve the bioactivity of scFv, we constructed a plasmid to express scFv-linker-matrilin-6×His fusion proteins that could self-assemble into the scFv dimers by disulfide bonds in matrilin under non-reducing conditions. An immunoreactivity assay demonstrated that the scFv dimer could highly bind to CTGF in a concentration-dependent manner. The MTT and EdU assay results revealed that CTGF (≥10 ng/mL) promoted the proliferation of ASM cells, and this effect was inhibited when the cells were treated with anti-CTGF scFv dimer. The western blot analysis results showed that increased phosphorylation of Akt and mTOR induced by CTGF could be suppressed by this scFv dimer. Based on these findings, anti-CTGF scFv dimer may be a potential agent for the prevention of airway remodeling in asthma. PMID:25478966

FvVE1 Regulates Biosynthesis of Fumonisins and Fusarins in Fusarium verticillioides

PubMed Central

MYUNG, KYUNG; LI, SHAOJIE; BUTCHKO, ROBERT A.E.; BUSMAN, MARK; PROCTOR, ROBERT H; ABBAS, HAMED K.; CALVO, ANA M.

2009-01-01

The veA gene positively regulates sterigmatocystin production in Aspergillus nidulans and aflatoxin production in A. parasiticus and A. flavus. Whether veA homologs have a role in regulating secondary metabolism in other fungal genera is unknown. In this study, we examined the role of the veA homolog, FvVE1, on production of two mycotoxin families, fumonisins and fusarins, in the important corn pathogen F. verticillioides. We found that FvVE1 deletion completely suppressed fumonisin production on two natural substrates, corn and rice. Furthermore, our results revealed that FvVE1 is necessary for the expression of the pathway-specific regulatory gene FUM21 and structural genes in the fumonisin biosynthetic gene (FUM) cluster. FvVE1 deletion also blocked production of fusarins. The effects of FvVE1 deletion on the production of these toxins were found to be the same in two separate mating types. Our results strongly suggest that FvVE1 play an important role in regulating mycotoxin production in F. verticillioides. PMID:19382792

Relationships between parenting style, feeding style and feeding practices and fruit and vegetable consumption in early childhood.

PubMed

Blissett, Jackie

2011-12-01

Despite substantial evidence suggesting that a diet high in fruit and vegetables (FV) is associated with reduced risk of cancer, only 21% of children in the UK consume the recommended 5 portions of fruit or vegetables a day. This review examines the role of parenting style, feeding style and feeding practices in FV consumption in early childhood. Whilst inconsistencies in concepts and terminology cloud this literature, overall the evidence suggests that the context of an authoritative parenting and feeding style is associated with better FV consumption in the childhood years. This context is typified by emotional warmth but high expectations for children's dietary adequacy and behaviour, accompanied by specific feeding practices such as modeling consumption of FV, making FV available within the home, covertly restricting unhealthy alternative snack foods, and encouraging children to try FV. Further longitudinal and intervention studies are required to determine the efficacy of modification of parenting style and feeding practice on children's FV intake. Copyright © 2011 Elsevier Ltd. All rights reserved.

Highly efficient recovery of functional single-chain Fv fragments from inclusion bodies overexpressed in Escherichia coli by controlled introduction of oxidizing reagent--application to a human single-chain Fv fragment.

PubMed

Tsumoto, K; Shinoki, K; Kondo, H; Uchikawa, M; Juji, T; Kumagai, I

1998-10-01

An improved and efficient refolding system for a single-chain antibody fragment (scFv) from inclusion bodies expressed in Escherichia coli was developed. Stepwise removal of denaturing reagent and controlled addition of oxidizing reagent were found to be the most effective conditions to achieve for almost complete recovery of functional monomeric scFv from inclusion bodies. Adding L-arginine to the refolding solution also increased the yield of refolded functional scFv. The single-chain Fv fragments of both a mouse anti-lysozyme monoclonal antibody, HyHEL10, and a human monoclonal antibody against the D antigen of the Rh blood group, D10, in solubilized inclusion bodies could be refolded under these conditions with yields of up to 95%. The refolding procedures developed in this study will contribute to providing a stable supply of large amounts of human single-chain Fv fragments.

Prediction of Winter Storm Tracks and Intensities Using the GFDL fvGFS Model

NASA Astrophysics Data System (ADS)

Rees, S.; Boaggio, K.; Marchok, T.; Morin, M.; Lin, S. J.

2017-12-01

The GFDL Finite-Volume Cubed-Sphere Dynamical core (FV3) is coupled to a modified version of the Global Forecast System (GFS) physics and initial conditions, to form the fvGFS model. This model is similar to the one being implemented as the next-generation operational weather model for the NWS, which is also FV3-powered. Much work has been done to verify fvGFS tropical cyclone prediction, but little has been done to verify winter storm prediction. These costly and dangerous storms impact parts of the U.S. every year. To verify winter storms we ran the NCEP operational cyclone tracker, developed at GFDL, on semi-real-time 13 km horizontal resolution fvGFS forecasts. We have found that fvGFS compares well to the operational GFS in storm track and intensity, though often predicts slightly higher intensities. This presentation will show the track and intensity verification from the past two winter seasons and explore possible reasons for bias.

Efficient refolding and immobilization of PMMA-tag-fused single-chain Fv antibodies for sensitive immunological detection on a PMMA plate.

PubMed

Kumada, Yoichi; Ishikawa, Yasuyuki; Fujiwara, Yusuke; Takeda, Rui; Miyamoto, Ryosuke; Niwa, Daisuke; Momose, Shun; Kang, Bongmun; Kishimoto, Michimasa

2014-09-01

In this study, we investigated the efficient refolding and site-specific immobilization of single-chain variable fragments (scFvs) genetically fused with a poly(methylmethacrylate)-binding peptide (PMMA-tag). According to the results of an aggregation test of a scFv-PM in the presence of 0.5 M urea, aggregation was hardly detectable at a weak-alkaline pH (8.5) with lower concentrations of NaCl. Consequently, more than 93% recovery of the anti-RNase scFv-PM model was attained, when it was refolded by dialysis against 50 mM TAPS (pH8.5). These results suggested that the apparent isoelectric point (pI) of a target scFv was decreased to a great extent by the genetic fusion of a PMMA-tag containing 5 acidic amino acids, and, thus, the solubility of the scFv-PM in its semi-denatured form was considerably improved. We also designed alternative peptide-tags composed of plural aspartic acid residues (D5, D10 and D15-tags) to decrease the apparent pI value of the fusion protein. As a consequence, scFv-D5, scFv-D10 and scFv-D15 were also efficiently refolded with yields of more than 95%. It is noteworthy that even scFv-PS-D15, which had both a positively charged polystyrene-binding peptide (PS-tag) and a negatively charged D15-tag, was serially connected at the C-terminal region of scFvs, and also refolded with a yield of 96.1%. These results clearly indicate that controlling the apparent pI value of scFvs by the fusion of oligo-peptides composed of acidic amino acids at the C-terminus resulted in a high degree of recovery via dialysis refolding. According to the results of a sandwich ELISA using scFv-PMs, scFv-D15 and scFv-PS-D15 as ligands, high antigen-binding signals were detected from both the PMMA and phi-PS plates immobilized with scFv-PMs. Furthermore, the high antigen-binding activity of scFv-PMs was maintained in an adsorption state when it was immobilized on the surface of not only PMMA, but also hydrophilic PS (phi-PS) and polycarbonate (PC). These results strongly suggested that a PMMA-tag introduced at the C-terminus of scFvs preferably recognizes ester and/or carboxyl groups exposed on the surface of plastics. The scFv-PM developed in the present study has advantages such as being a ligand antibody, compared with whole Ab and the conventional PS-tag-fused scFvs (scFv-PS), and, thus, it is considerably useful in a sandwich ELISA as well as in various immuno-detection and immuno-separation systems. Copyright © 2014 Elsevier B.V. All rights reserved.

Fruit and vegetable purchasing patterns and preferences in South Delhi

PubMed Central

FINZER, LAUREN E.; AJAY, VAMADEVAN S.; ALI, MOHAMMED K.; SHIVASHANKAR, ROOPA; GOENKA, SHIFALIKA; PILLAI, DIVYA S.; KHANDELWAL, SHWETA; TANDON, NIKHIL; REDDY, K. SRINATH; NARAYAN, K.M. VENKAT; PRABHAKARAN, DORAIRAJ

2017-01-01

This study examines associations between consumer characteristics, beliefs, and preferences and fruit and vegetable (FV) purchasing and intake in South Delhi, India. Home interviews were conducted with 245 households, using a structured questionnaire to assess FV consumption and purchasing frequency, spending, place of purchase, mode of travel, knowledge and attitudes towards organics, and beliefs about barriers to FV consumption. In-depth interviews with 62 experts and key informants validated survey findings that street vendors and markets are currently the dominant source of FV in South Delhi and that affordability, not accessibility, is the main barrier to increasing FV intake. PMID:23282188

Healthier side dishes at restaurants: an analysis of children’s perspectives, menu content, and energy impacts

PubMed Central

2014-01-01

Background Children consume restaurant-prepared foods at high rates, suggesting that interventions and policies targeting consumption of these foods have the potential to improve diet quality and attenuate excess energy intake. One approach to encouraging healthier dietary intake in restaurants is to offer fruits and vegetables (FV) as side dishes, as opposed to traditional, energy-dense accompaniments like French fries. The aims of the current study were to examine: children's views about healthier side dishes at restaurants; current side dish offerings on children's menus at leading restaurants; and potential energy reductions when substituting FV side dishes in place of French fries. Methods To investigate children’s attitudes, a survey was administered to a nationally representative sample of U.S. 8- to 18-year-olds (n = 1178). To examine current side dish offerings, children's menus from leading quick service (QSR; n = 10) and full service restaurant chains (FSR; n = 10) were analyzed. Energy reductions that could result from substituting commonly-offered FV side dishes for French fries were estimated using nutrition information corresponding to the children's menu items. Results Two-thirds of children reported that they would not feel negatively about receiving FV sides instead of French fries with kids' meals. Liking/taste was the most common reason that children gave to explain their attitudes about FV side dishes. Nearly all restaurants offered at least 1 FV side dish option, but at most restaurants (60% of QSR; 70% of FSR), FV sides were never served by default. Substituting FV side dishes for French fries yielded an average estimated energy reduction of at least 170 calories. Conclusions Results highlight some healthy trends in the restaurant context, including the majority of children reporting non-negative attitudes about FV side dishes and the consistent availability of FV side dish options at leading QSR and FSR. Yet the minority of restaurants offer these FV sides by default. Promoting creative, appealing FV side dishes can result in healthier, less energy-dense meals for children. Substituting or displacing energy-dense default side dishes with such FV dishes show promise as part of continued, comprehensive efforts to increase the healthfulness of meals consumed by children in restaurant settings. PMID:24996545

Associations between socioeconomic, parental and home environment factors and fruit and vegetable consumption of children in grades five and six in British Columbia, Canada.

PubMed

Attorp, Adrienne; Scott, Jenny E; Yew, Ann C; Rhodes, Ryan E; Barr, Susan I; Naylor, Patti-Jean

2014-02-11

Regular fruit and vegetable (FV) consumption has been associated with reduced chronic disease risk. Evidence from adults shows a social gradient in FV consumption. Evidence from pre-adolescent children varies and there is little Canadian data. This study assessed the FV intake of school children in British Columbia (BC), Canada to determine whether socio-economic status (SES), parental and the home environment factors were related to FV consumption. As part of the BC School Fruit and Vegetable Nutrition Program, 773 British Columbia fifth-and sixth-grade school children (Mean age 11.3 years; range 10.3-12.5) and their parents were surveyed to determine FV consumption and overall dietary intake. Students completed a web-based 24-hour dietary food recall, and a student measure of socio-economic status (The Family Affluence Scale). Parents completed a self-administered survey about their education, income, home environment and perceptions of their neighbourhood and children's eating habits. Correlations and multiple regression analyses were used to examine the association between SES, parental and home environment factors and FV consumption. Approximately 85.8% of children in this study failed to meet minimum Canadian guidelines for FV intake (6 servings). Parent income and education were not significantly associated with child FV consumption but were associated with each other, child-reported family affluence, neighbourhood environment, access to FV, and eating at the table or in front of the television. Significant positive associations were found between FV consumption and child-reported family affluence, meal-time habits, neighbourhood environment and parent perceptions of the healthiness of their child's diet; however, these correlations were weak (ranging from .089-.115). Multiple regression analysis showed that only child-reported family affluence significantly predicted FV consumption (std-β = 0.096 95% CI = 0.01 to 0.27). The majority of children in our study were not meeting guidelines for FV intake irrespective of SES, parent perceptions or home environment, making this a population wide concern. An almost trivial socio-economic gradient was observed for the child-reported SES measure only. These results are consistent with several other studies of children. Longitudinal research is needed to further explore individual and social factors associated with FV consumption in childhood and their development over time.

Development of a microplate coagulation assay for Factor V in human plasma

PubMed Central

2011-01-01

Background Factor V (FV) in its activated form, FVa, is a critical regulator of thrombin generation during fibrin clot formation. There is a need of a simple, fast, and inexpensive microplate-based coagulation assay to measure the functional activity of FV in human plasma. The objective of this study was to develop a microplate-based assay that measures FV coagulation activity during clot formation in human plasma, which is currently not available. Methods The FV assay requires a kinetic microplate reader to measure the change in absorbance at 405nm during fibrin formation in human plasma. The FV assay accurately measures the time, initial rate, and extent of fibrin clot formation in human plasma. Results The FV microplate assay is simple, fast, economical, sensitive to approx 24-80pM, and multiple samples may be analyzed simultaneously. All the required materials are commercially available. Standard curves of time or initial rate of fibrin clot formation vs FV activity in the 1-stage assay (Without activation by thrombin) may be used to measure FV activity in samples of human plasma. The assay was used to demonstrate that in nine patients with disseminated intravascular coagulation (DIC), the FV 1-stage, 2-stage (With activation by thrombin), and total (2-stage activity - 1-stage activity) activities were decreased, on average, by approximately 54%, 44%, and 42%, respectively, from prolonged clot times when compared to normal pooled human reference plasma (NHP). The results indicate that the FV in the DIC patient plasmas supported both a delayed and slower rate of fibrin clot formation compared with NHP; however, the extent of fibrin clot formation in the DIC patients remained largely unchanged from that observed with NHP. Conclusions The FV microplate assay may be easily adapted to measure the activity of any coagulation factor using the appropriate factor-deficient plasma and clot initiating reagent. The microplate assay will find use in both research and clinical laboratories to provide measurement of the functional coagulation activity of FV in human plasma. PMID:21711555

Cell growth inhibition and apoptotic effects of a specific anti-RTFscFv antibody on prostate cancer, but not glioblastoma, cells

PubMed Central

Nejatollahi, Foroogh; Bayat, Payam; Moazen, Bahareh

2017-01-01

Background: Single chain antibody (scFv) has shown interesting results in cancer immunotargeting approaches, due to its advantages over monoclonal antibodies. Regeneration and tolerance factor (RTF) is one of the most important regulators of extracellular and intracellular pH in eukaryotic cells. In this study, the inhibitory effects of a specific anti-RTF scFv were investigated and compared between three types of prostate cancer and two types of glioblastoma cells.  Methods: A phage antibody display library of scFv was used to select specific scFvs against RTF using panning process. The reactivity of a selected scFv was assessed by phage ELISA. The anti-proliferative and apoptotic effects of the antibody on prostate cancer (PC-3, Du-145 and LNCaP) and glioblastoma (U-87 MG and A-172) cell lines were investigated by MTT and Annexin V/PI assays.  Results: A specific scFv with frequency 35% was selected against RTF epitope. This significantly inhibited the proliferation of the prostate cells after 24 h. The percentages of cell viability (using 1000 scFv/cell) were 52, 61 and 73% for PC-3, Du-145 and LNCaP cells, respectively, compared to untreated cells. The antibody (1000 scFv/cell) induced apoptosis at 50, 40 and 25% in PC-3, Du-145 and LNCaP cells, respectively. No growth inhibition and apoptotic induction was detected for U-87 and A172 glioblastoma cells.  Conclusions: Anti-RTFscFv significantly reduced the proliferation of the prostate cancer cells. The inhibition of cell growth and apoptotic induction effects in PC-3 cells were greater than Du-145 and LNCaP cells. This might be due to higher expression of RTF antigen in PC-3 cells and/or better accessibility of RTF to scFv antibody. The resistance of glioblastoma cells to anti-RTF scFv offers the existence of mechanism(s) that abrogate the inhibitory effect(s) of the antibody to RTF. The results suggest that the selected anti-RTF scFv antibody could be an effective new alternative for prostate cancer immunotherapy. PMID:28491282

A Supermarket Double-Dollar Incentive Program Increases Purchases of Fresh Fruits and Vegetables Among Low-Income Families With Children: The Healthy Double Study.

PubMed

Polacsek, Michele; Moran, Alyssa; Thorndike, Anne N; Boulos, Rebecca; Franckle, Rebecca L; Greene, Julie C; Blue, Dan J; Block, Jason P; Rimm, Eric B

2018-03-01

To carry out a pilot study to determine whether a supermarket double-dollar fruit and vegetable (F&V) incentive increases F&V purchases among low-income families. Randomized controlled design. Purchases were tracked using a loyalty card that provided participants with a 5% discount on all purchases during a 3-month baseline period followed by the 4-month intervention. A supermarket in a low-income rural Maine community. A total of 401 low-income and Supplemental Nutrition Assistance Program (SNAP) supermarket customers. Same-day coupon at checkout for half-off eligible fresh, frozen, or canned F&V over 4 months. Weekly spending in dollars on eligible F&V. A linear model with random intercepts accounted for repeated transactions by individuals to estimate change in F&V spending per week from baseline to intervention. Secondary analyses examined changes among SNAP-eligible participants. Coupons were redeemed among 53% of eligible baskets. Total weekly F&V spending increased in the intervention arm compared with control ($1.83; 95% confidence interval [CI], $0.29 to $3.88). The largest increase was for fresh F&V ($1.97; 95% CI, $0.49 to $3.44). Secondary analyses revealed greater increases in F&V spending among SNAP-eligible participants who redeemed coupons ($5.14; 95% CI, $1.93 to $8.34) than among non-SNAP eligible participants who redeemed coupons ($3.88; 95% CI, $1.67 to $6.08). A double-dollar pricing incentive increased F&V spending in a low-income community despite the moderate uptake of the coupon redemption. Customers who were eligible for SNAP saw the greatest F&V spending increases. Financial incentives for F&V are an effective strategy for food assistance programs to increase healthy purchases and improve dietary intake in low-income families. Copyright © 2017 Society for Nutrition Education and Behavior. Published by Elsevier Inc. All rights reserved.

Do We Produce Enough Fruits and Vegetables to Meet Global Health Need?

PubMed Central

Siegel, Karen R.; Ali, Mohammed K.; Srinivasiah, Adithi; Nugent, Rachel A.; Narayan, K. M. Venkat

2014-01-01

Background Low fruit and vegetable (FV) intake is a leading risk factor for chronic disease globally, but much of the world’s population does not consume the recommended servings of FV daily. It remains unknown whether global supply of FV is sufficient to meet current and growing population needs. We sought to determine whether supply of FV is sufficient to meet current and growing population needs, globally and in individual countries. Methods and Findings We used global data on agricultural production and population size to compare supply of FV in 2009 with population need, globally and in individual countries. We found that the global supply of FV falls, on average, 22% short of population need according to nutrition recommendations (supply:need ratio: 0.78 [Range: 0.05–2.01]). This ratio varies widely by country income level, with a median supply:need ratio of 0.42 and 1.02 in low-income and high-income countries, respectively. A sensitivity analysis accounting for need-side food wastage showed similar insufficiency, to a slightly greater extent (global supply:need ratio: 0.66, varying from 0.37 [low-income countries] to 0.77 [high-income countries]). Using agricultural production and population projections, we also estimated supply and need for FV for 2025 and 2050. Assuming medium fertility and projected growth in agricultural production, the global supply:need ratio for FV increases slightly to 0.81 by 2025 and to 0.88 by 2050, with similar patterns seen across country income levels. In a sensitivity analysis assuming no change from current levels of FV production, the global supply:need ratio for FV decreases to 0.66 by 2025 and to 0.57 by 2050. Conclusion The global nutrition and agricultural communities need to find innovative ways to increase FV production and consumption to meet population health needs, particularly in low-income countries. PMID:25099121

Time to address continued poor vegetable intake in Australia for prevention of chronic disease.

PubMed

Chapman, Kathryn; Havill, Michelle; Watson, Wendy L; Wellard, Lyndal; Hughes, Clare; Bauman, Adrian; Allman-Farinelli, Margaret

2016-12-01

Australian and most international Dietary Guidelines recommend people consume more fruits and vegetables (F&V) to maintain a healthy weight and reduce chronic disease risk. Previous Australian and international surveys have shown sub-optimal consumption of F&V. This study aimed to assess adults' F&V consumption, knowledge of recommended servings, readiness to change, barriers/enabling factors, so that this knowledge might be used for campaigns that support improved consumption. An online survey of a representative sample of adults living in New South Wales, Australia (n = 2474) measuring self-reported F&V consumption; attitudes towards F&V consumption; stage of change for increasing F&V; barriers to consumption; and knowledge of cancer-health benefits. F&V consumption was below recommendations, with vegetable consumption notably low. Only 10% of participants ate at least five servings of vegetables/day (median intake was two daily servings), and 57% consumed two servings fruit/day. There was poor recognition that intake of vegetables was inadequate and this was a barrier to improving vegetable consumption; with preferences for other foods, habit and cost also important barriers. Key barriers to increasing fruit intake were habit, preferences for other foods, perishability, and cost. For vegetable consumption, 49% of participants were in the pre-contemplation stage of change, whereas for fruits 56% were in the action/maintenance stage. Sixty-four percent of respondents believed that eating F&V would protect against cancer, with 56% reporting they thought not eating enough F&V would cause cancer. Understanding what motivates and prevents people from consuming F&V is important for developing effective health promotion programs. Similar to previous surveys, there has been little shift in F&V consumption. Social marketing campaigns have been shown to improve health-related behaviours, and this study may assist in identifying audience segmentation for better targeted campaigns. Copyright © 2016 Elsevier Ltd. All rights reserved.

Potent inhibition of OKT3-induced T cell proliferation and suppression of CD147 cell surface expression in HeLa cells by scFv-M6-1B9.

PubMed

Intasai, Nutjeera; Tragoolpua, Khajornsak; Pingmuang, Prakitnavin; Khunkaewla, Panida; Moonsom, Seangdeun; Kasinrerk, Watchara; Lieber, André; Tayapiwatana, Chatchai

2008-01-01

CD147, a multifunctional type I transmembrane glycoprotein, has been implicated in various physiological and pathological processes. It is involved in signal transduction pathways and also plays a crucial role in the invasive and metastatic activity of malignant tumor cells. Diminished expression of this molecule has been shown to be beneficial in suppression of tumor progression. In a previous study, we generated and characterized a recombinant antibody fragment, scFv, which reacted specifically to CD147. In the present study, we further investigated the biological properties, function and the effect of generated scFv on CD147 expression. The in vitro study showed that soluble scFv-M6-1B9 produced from E. coli HB2151 bound to CD147 surface molecule and inhibited OKT3-induced T cell proliferation. Furthermore, soluble lysate of scFv-M6-1B9 from 293A cells, transduced with a scFv-M6-1B9 expressing adenovirus vector, recognized both recombinant and native CD147. These results indicate that scFv-M6-1B9 binds with high efficiency and specificity. Importantly, scFv-M6-1B9 intrabody reduced the expression of CD147 on the cell surface of HeLa cells suggesting that scFv-M6-1B9 is biologically active. In conclusion, our present study demonstrated that scFv-M6-1B9 has a great potential to target both the intracellular and the extracellular CD147. The generated scFv-M6-1B9 may be an effective agent to clarify the cellular function of CD147 and may aid in efforts to develop a novel treatment in various human carcinomas.

Newly-developed, forward-viewing echoendoscope: a comparative pilot study to the standard echoendoscope in the imaging of abdominal organs and feasibility of endoscopic ultrasound-guided interventions.

PubMed

Iwashita, Takuji; Nakai, Yousuke; Lee, John G; Park, Do Hyun; Muthusamy, V Raman; Chang, Kenneth J

2012-02-01

Multiple diagnostic and therapeutic endoscopic ultrasound (EUS) procedures have been widely performed using a standard oblique-viewing (OV) curvilinear array (CLA) echoendoscope. Recently, a new, forward-viewing (FV) CLA was developed, with the advantages of improved endoscopic viewing and manipulation of devices. However, the FV-CLA echoendoscope has a narrower ultrasound scanning field, and lacks an elevator, which might represent obstacles for clinical use. The aim of this study was to compare the FV-CLA echoendoscope to the OV-CLA echoendoscope for EUS imaging of abdominal organs, and to assess the feasibility of EUS-guided interventions using the FV-CLA echoendoscope. EUS examinations were first performed and recorded using the OV-CLA echoendoscope, followed immediately by the FV-CLA echoendoscope. Video recordings were then assessed by two independent endosonographers in a blinded fashion. The EUS visualization and image quality of specific abdominal organs/structures were scored. Any indicated fine-needle aspiration (FNA) or intervention was performed using the FV-CLA echoendoscope, with the OV-CLA echoendoscope as salvage upon failure. A total of 21 patients were examined in the study. Both echoendoscopes had similar visualization and image quality for all organs/structures, except the common hepatic duct (CHD), which was seen significantly better with the FV-CLA echoendoscope. EUS interventions were conducted in eight patients, including FNA of pancreatic mass (3), pancreatic cyst (3), and cystgastrostomy (2). The FV-CLA echoendoscope was successful in seven patients. One failed FNA of the pancreatic head cyst was salvaged using the OV-CLA echoendoscope. There were no differences between the FV-CLA echoendoscope and the OV-CLA echoendoscope in visualization or image quality on upper EUS, except for the superior image quality of CHD using the FV-CLA echoendoscope. Therefore, the disadvantages of the FV-CLA echoendoscope appear minimal in light of the potential advantages. © 2011 Journal of Gastroenterology and Hepatology Foundation and Blackwell Publishing Asia Pty Ltd.

Development of potential breeder-friendly markers for the I gene using bulked segregant analysis and whole-genome sequencig

USDA-ARS?s Scientific Manuscript database

Bean common mosaic virus (BCMV) and its related necrotic species Bean common mosaic necrosis virus (BCMNV), are the most prevalent potyviruses in common bean (Phaseolus vulgaris). The dominant I gene confers immunity or temperature-dependent hypersensitive response to most strains of BCMV, and tempe...

Consumer segmentation based on the level and structure of fruit and vegetable intake: an empirical evidence for US adults from the National Health and Nutrition Examination Survey (NHANES) 2005-2006.

PubMed

Demydas, Tetyana

2011-06-01

To identify consumption patterns of fruit and vegetables within a representative sample of US adults with a focus on degree of produce processing and to explore sociodemographic, lifestyle and nutritional profiles associated with these patterns. Cross-sectional analysis. Fruit and vegetable (F&V) consumption data were collected using two non-consecutive 24 h recalls. For the purpose of the study, F&V intakes were aggregated into seven subgroups indicating degree of processing, which afterwards were used as inputs into cluster analysis. The 2005-2006 National Health and Nutrition Examination Survey. The sample consisted of 2444 adults aged 20-59 years. Total average F&V intake of the adults was below the recommended level. Thereby, 20 % of the respondents consumed fruit only in the form of juice. Three F&V consumption patterns were identified: 'low-intake F&V consumers' (74 % of respondents), 'consumers of healthier F&V options' (13 %) and 'intensive fruit juice consumers' (13 %). These groups differed markedly in terms of their sociodemographic, lifestyle and health characteristics, such as gender, age, race/ethnicity, education, smoking, weight status, etc. Differences in nutrient profiles were also found, with the 'consumers of healthier F&V options' showing better nutritional quality compared with other clusters. Only a small share of US adults combines high F&V intakes with healthier F&V options that lead to a better nutritional profile. This raises discussion about a need to deliver more specific F&V promotion messages, including advice on healthier preparation methods, especially for the specific population groups.

Synergistic capture of Clostridium botulinum Type A neurotoxin by scFv antibodies to novel epitopes

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

Gray, Sean A.; Barr, John R.; Kalb, Suzanne R.

2011-10-01

A non-immune library of human single chain fragment variable (scFv) antibodies displayed on Saccharomyces cerevisiae was screened for binding to the Clostridium botulinum neurotoxin serotype A binding domain [BoNT/A (Hc)] with the goal of identifying scFv to novel epitopes. To do this, an antibody-mediated labeling strategy was used in which antigen-binding yeast clones were selected after labeling with previously characterized monoclonal antibodies (MAbs) specific to the Hc. Twenty unique scFv clones were isolated that bound Hc. Of these, three also bound to full-length BoNT/A toxin complex with affinities ranging from 5 nM to 170 nM. Epitope binning showed that themore » three unique clones recognized at least two epitopes that were distinct from one another and from the detection MAbs. After production in E. coli, the scFv were coupled to magnetic particles and tested for their ability to capture BoNT/A holotoxin using an Endopep-MS assay. In this assay, toxin captured by scFv coated magnetic particles was detected by incubation of the complex with a peptide containing a BoNT/A-specific cleavage sequence. Mass spectrometry was used to detect the ratio of intact peptide to cleavage products as evidence for toxin capture. When tested individually, each of the scFv showed a weak positive Endopep-MS result. However, when the particles were coated with all three scFv simultaneously, they exhibited significantly higher Endopep-MS activity, consistent with synergistic binding. These results demonstrate novel approaches toward the isolation and characterization of scFv antibodies specific to unlabeled antigen. They also provide evidence that distinct scFv antibodies can work synergistically to increase the efficiency of antigen capture onto a solid support.« less

How much is '5-a-day'? A qualitative investigation into consumer understanding of fruit and vegetable intake guidelines.

PubMed

Rooney, C; McKinley, M C; Appleton, K M; Young, I S; McGrath, A J; Draffin, C R; Hamill, L L; Woodside, J V

2017-02-01

Despite the known health benefits of fruit and vegetables (FV), population intakes remain low. One potential contributing factor may be a lack of understanding surrounding recommended intakes. The present study aimed to explore the understanding of FV intake guidelines among a sample of low FV consumers. Six semi-structured focus groups were held with low FV consumers (n = 28, age range 19-55 years). Focus groups were recorded digitally, transcribed verbatim and analysed thematically using nvivo (QSR International, Melbourne, Australia) to manage the coded data. Participants also completed a short questionnaire assessing knowledge on FV intake guidelines. Descriptive statistics were used to analyse responses. The discussions highlighted that, although participants were aware of FV intake guidelines, they lacked clarity with regard to the meaning of the '5-a-day' message, including what foods are included in the guideline, as well as what constitutes a portion of FV. There was also a sense of confusion surrounding the concept of achieving variety with regard to FV intake. The sample highlighted a lack of previous education on FV portion sizes and put forward suggestions for improving knowledge, including increased information on food packaging and through health campaigns. Questionnaire findings were generally congruent with the qualitative findings, showing high awareness of the '5-a-day' message but a lack of knowledge surrounding FV portion sizes. Future public health campaigns should consider how best to address the gaps in knowledge identified in the present study, and incorporate evaluations that will allow the impact of future initiatives on knowledge, and ultimately behaviour, to be investigated. © 2016 The British Dietetic Association Ltd.

Factors within the family environment such as parents' dietary habits and fruit and vegetable availability have the greatest influence on fruit and vegetable consumption by Polish children.

PubMed

Wolnicka, Katarzyna; Taraszewska, Anna Małgorzata; Jaczewska-Schuetz, Joanna; Jarosz, Mirosław

2015-10-01

To identify determinants of fruit and vegetable (F&V) consumption among school-aged children. A survey study was conducted in October 2010. The questionnaire contained questions concerning social and demographic data, lifestyle and dietary habits, particularly the frequency of F&V consumption, availability of F&V and knowledge about recommended amounts of F&V intake. Polish primary schools. Children (n 1255) aged 9 years from randomly selected primary schools and their parents. The children's consumption of fruit and of vegetables was influenced by the fruit consumption and vegetable consumption of their parents (r=0·333 and r=0·273, respectively; P=0·001), parents encouraging their children to eat F&V (r=0·259 and r=0·271, respectively; P=0·001), giving children F&V to take to school (r=0·338 and r=0·321, respectively; P=0·001) and the availability of F&V at home (r=0·200 and r=0·296, respectively; P=0·001). Parental education influenced only the frequency of fruit consumption (r=0·074; P=0·01). A correlation between parents' knowledge of the recommended intakes and the frequency of vegetable and fruit consumption by children was noticed (r=0·258 and r=0·192, respectively, P=0·001). Factors within the family environment such as parents' dietary habits and F&V availability had the greatest influence on the F&V consumption by children. Educational activities aimed at parents are crucial to increase the consumption of F&V among children.

Impact of the revised Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) food package policy on fruit and vegetable prices.

PubMed

Zenk, Shannon N; Powell, Lisa M; Odoms-Young, Angela M; Krauss, Ramona; Fitzgibbon, Marian L; Block, Daniel; Campbell, Richard T

2014-02-01

Obesity is generally inversely related to income among women in the United States. Less access to healthy foods is one way lower income can influence dietary behaviors and body weight. Federal food assistance programs, such as the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC), are an important source of healthy food for low-income populations. In 2009, as part of a nationwide policy revision, WIC added a fruit and vegetable (F/V) voucher to WIC food packages. This quasi-experimental study determined whether F/V prices at stores authorized to accept WIC (ie, WIC vendors) decreased after the policy revision in seven Illinois counties. It also examined cross-sectional F/V price variations by store type and neighborhood characteristics. Two pre-policy observations were conducted in 2008 and 2009; one post-policy observation was conducted in 2010. Small pre- to post-policy reductions in some F/V prices were found, particularly for canned fruit and frozen vegetables at small stores. Compared with chain supermarkets, mass merchandise stores had lower prices for fresh F/V and frozen F/V and small stores and non-chain supermarkets had higher canned and frozen F/V prices, but lower fresh F/V prices. Limited price differences were found across neighborhoods, although canned vegetables were more expensive in neighborhoods with higher concentrations of either Hispanics or blacks and fresh F/V prices were lower in neighborhoods with more Hispanics. Results suggest the WIC policy revision contributed to modest reductions in F/V prices. WIC participants' purchasing power can differ depending on the type and neighborhood of the WIC vendor used. Copyright © 2014 Academy of Nutrition and Dietetics. Published by Elsevier Inc. All rights reserved.

Prevalence of Factor V Genetic Variants Associated With Indian APCR Contributing to Thrombotic Risk.

PubMed

Sharma, Amit; Bhakuni, Teena; Biswas, Arijit; Ranjan, Ravi; Kumar, Ravi; Kishore, Kamal; Mahapatra, Manoranjan; Jairajpuri, Mohamad Aman; Saxena, Renu

2017-09-01

Phenotypic resistance to activated protein C (APC) is a complex mechanism associated with increased thrombosis risk. Activated protein C resistance (APCR) is mainly influenced by FV Leiden mutation, and various other single nucleotide polymorphisms (SNPs) in FV gene are known to be associated with APCR. The aim of present study was to investigate the incidence and assess possible mechanisms of APCR in Indian patients with deep vein thrombosis (DVT). Three hundred and ten Doppler-proven patients with DVT were screened for APCR, and 50 APCR positive patients and 50 controls were typed for FV Leiden , Hong Kong, Cambridge, HR2 haplotype, Glu666Asp, Ala485Lys, and Liverpool using either polymerase chain reaction (PCR)-restriction fragment length polymorphism or allele specific PCR. FV Leiden was commonest cause of APCR (50%) in Indian patients with DVT being statistically significant ( P = .001) compared to controls. FV Liverpool, FV Glu666Asp and FV Ala485Lys were studied for the first time in Indian population. FV Liverpool, FV Glu666Asp, Hong Kong, and Cambridge were found to be absent. High frequency of Ala485Lys in patients shows that it might be a risk factor contributing to APCR in Indian patients with DVT. HR2 haplotype was not associated with APCR; however, presence of homozygous HR2 haplotype in patients only indicates the role it might play in Indian APCR population. In conclusion, contribution of FV Leiden causing APCR in Indian population is not as strong as previously reported in Western countries. The presence of other SNPs observed in the present study requires such studies on larger sample size to understand the molecular basis of defect.

FV-100 versus valacyclovir for the prevention of post-herpetic neuralgia and the treatment of acute herpes zoster-associated pain: A randomized-controlled trial.

PubMed

Tyring, Stephen K; Lee, Patricia; Hill, Gordon T; Silverfield, Joel C; Moore, Angela Yen; Matkovits, Theresa; Sullivan-Bolyai, John

2017-07-01

This prospective, parallel-group, randomized, double-blind, multicenter study compared the efficacy and safety of FV-100 with valacyclovir for reducing pain associated with acute herpes zoster (HZ). Patients, ≥50 years of age, diagnosed with HZ within 72 h of lesion appearance who had HZ-associated pain, were randomized 1:1:1 to a 7-day course of either FV-100 200 mg QD (n = 117), FV-100 400 mg QD (n = 116), or valacyclovir 1000 mg TID (n =117). Efficacy was evaluated on the basis of the burden of illness (BOI; Zoster Brief Pain Inventory scores); incidence and duration of clinically significant pain (CSP); pain scores; incidence and severity of post-herpetic neuralgia (PHN); and times to full lesion crusting and to lesion healing. Safety was evaluated on the basis of adverse event (AE)/SAE profiles, changes in laboratory and vital signs values, and results of electrocardiograms. The burden of illness scores for pain through 30 days were 114.5, 110.3, and 118.0 for FV-100 200 mg, FV-100 400 mg, and valacyclovir 3000 mg, respectively. The incidences of PHN at 90 days for FV-100 200 mg, FV-100 400 mg, and valacyclovir 3000 mg were 17.8%, 12.4%, and 20.2%, respectively. Adverse event and SAE profiles of the two FV-100 and the valacyclovir groups were similar and no untoward signals or trends were evident. These results demonstrate a potential for FV-100 as an antiviral for the treatment of shingles that could both reduce the pain burden of the acute episode and reduce the incidence of PHN compared with available treatments. © 2016 Wiley Periodicals, Inc.

[Construction and screening of phage antibody libraries against epidermal growth factor receptor and soluble expression of single chain Fv].

PubMed

Sheng, Wei-Jin; Miao, Qing-Fang; Zhen, Yong-Su

2009-06-01

Recent studies have shown that epidermal growth factor receptor (EGFR) is an important target for cancer therapy. The present study prepared single chain Fv (scFv) directed against EGFR. Balb/c mice were immunized by human carcinoma A431 cells, and total RNA of the splenic cells was extracted. VH and VL gene fragments were amplified by RT-PCR and further joined into scFv gene with a linker, then scFv gene fragments were ligated into the phagemid vector pCANTAB 5E. The phagemid containing scFv were transformed into electro-competent E. coli TG1 cells. The recombinant phage antibody library was constructed through rescuing the transformed cells with help phage M13K07. The specified recombinant phages were enriched through 5 rounds of affinity panning and the anti-EGFR phage scFv clones were screened and identified with ELISA. A total of 48 clones from the library were selected randomly and 45 clones were identified positive. After infecting E. coli HB2151 cells with one positive clone, soluble recombinant antibodies about 27 kD were produced and located in the periplasm and the supernatant. The result of sequencing showed that the scFv gene was 768 bp, which encoded 256 amino acid residues. VH and VL including 3 CDRs and 4 FRs, respectively, were all homologous to mouse Ig. The soluble scFv showed the specific binding activity to purified EGFR and EGFR located in carcinoma cell membrane. The successful preparation of anti-EGFR scFv will provide an EGFR targeted molecule for the development of antibody-based drugs and biological therapy of cancer.

Impact of the Revised Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) Food Package Policy on Fruit and Vegetable Prices

PubMed Central

Zenk, Shannon N.; Powell, Lisa M.; Odoms-Young, Angela M.; Krauss, Ramona; Fitzgibbon, Marian L.; Block, Daniel; Campbell, Richard T.

2014-01-01

Obesity is generally inversely related to income among women in the United States. Less access to healthy foods is one way lower income can influence dietary behaviors and body weight. Federal food assistance programs, such as the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC), are an important source of healthy food for low-income populations. In 2009, as part of a nationwide policy revision, WIC added a fruit and vegetable (F/V) voucher to WIC food packages. This quasi-experimental study determined whether F/V prices at stores authorized to accept WIC (ie, WIC vendors) decreased after the policy revision in seven Illinois counties. It also examined cross-sectional F/V price variations by store type and neighborhood characteristics. Two pre-policy observations were conducted in 2008 and 2009; one post-policy observation was conducted in 2010. Small pre- to post-policy reductions in some F/V prices were found, particularly for canned fruit and frozen vegetables at small stores. Compared with chain supermarkets, mass merchandise stores had lower prices for fresh F/V and frozen F/V and small stores and non-chain supermarkets had higher canned and frozen F/V prices, but lower fresh F/V prices. Limited price differences were found across neighborhoods, although canned vegetables were more expensive in neighborhoods with higher concentrations of either Hispanics or blacks and fresh F/V prices were lower in neighborhoods with more Hispanics. Results suggest the WIC policy revision contributed to modest reductions in F/V prices. WIC participants’ purchasing power can differ depending on the type and neighborhood of the WIC vendor used. PMID:24183996

Identification and Transcript Analysis of the TCP Transcription Factors in the Diploid Woodland Strawberry Fragaria vesca

PubMed Central

Wei, Wei; Hu, Yang; Cui, Meng-Yuan; Han, Yong-Tao; Gao, Kuan; Feng, Jia-Yue

2016-01-01

Plant-specific TEOSINTE BRANCHED 1, CYCLOIDEA, and PROLIFERATING CELL FACTORS (TCP) transcription factors play versatile functions in multiple processes of plant growth and development. However, no systematic study has been performed in strawberry. In this study, 19 FvTCP genes were identified in the diploid woodland strawberry (Fragaria vesca) accession Heilongjiang-3. Phylogenetic analysis suggested that the FvTCP genes were classified into two main classes, with the second class further divided into two subclasses, which was supported by the exon-intron organizations and the conserved motif structures. Promoter analysis revealed various cis-acting elements related to growth and development, hormone and/or stress responses. We analyzed FvTCP gene transcript accumulation patterns in different tissues and fruit developmental stages. Among them, 12 FvTCP genes exhibited distinct tissue-specific transcript accumulation patterns. Eleven FvTCP genes were down-regulated in different fruit developmental stages, while five FvTCP genes were up-regulated. Transcripts of FvTCP genes also varied with different subcultural propagation periods and were induced by hormone treatments and biotic and abiotic stresses. Subcellular localization analysis showed that six FvTCP-GFP fusion proteins showed distinct localizations in Arabidopsis mesophyll protoplasts. Notably, transient over-expression of FvTCP9 in strawberry fruits dramatically affected the expression of a series of genes implicated in fruit development and ripening. Taken together, the present study may provide the basis for functional studies to reveal the role of this gene family in strawberry growth and development. PMID:28066489

Fruit and vegetable intake and smoking cessation.

PubMed

Poisson, T; Dallongeville, J; Evans, A; Ducimetierre, P; Amouyel, P; Yarnell, J; Bingham, A; Kee, F; Dauchet, L

2012-11-01

In cohort studies, fruit and vegetable (F&V) intake is associated with lower cardiovascular diseases (CVDs). Former smokers often have a higher F&V intake than current smokers. If a high intake of F&V precedes smoking cessation, the latter may explain the favorable association between F&V intake and CVD among smokers. The objective was to assess whether higher F&V intake precedes smoking cessation. The study population comprised 1056 male smokers from Lille (France) and Belfast (Northern Ireland) aged 50-59 years on inclusion in 1991. At baseline, participants completed self-administered questionnaires related to smoking habits, demographic, socioeconomic factors and diet. At the 10-year follow-up, smoking habits were assessed by mailed questionnaire. After 10 years, 590 out of 1056 smokers had quit smoking (70.7% of smoker in Lille and 37.8% in Belfast). After adjusting for center, consumption of F&V was associated with quitting (odds ratio (OR) for high versus low F&V intake: 1.73; 95% confidence interval (CI): (1.22-2.45); P-trend=0.002). After further adjustment for sociodemographic factors, body mass index and medical diet, the association was still statistically significant (OR: 1.59; 95% CI (1.12-2.27); P-trend = 0.01). In a model fully adjusted for age, smoking intensity, alcohol consumption and physical activity, the association was no longer significant (P = 0.14). Higher F&V intake precedes smoking cessation. Hence, smoking cessation could affect the causal interpretation of the association between F&V and CVD in smokers.

Psychometrics of the "Self-Efficacy Consumption of Fruit and Vegetables Scale" in African American women.

PubMed

Gittner, Lisaann S; Gittner, Kevin B

2017-08-01

Assess the psychometric properties of the Self-Efficacy Consumption of Fruit and Vegetable Scale (F/V scale) in African American women. Midwestern Health Maintenance Organization. 221 African American women age 40-65 with BMI≥30 MEASURES: F/V scale was compared to eating efficacy/availability subscale reported on the WEL and mean micronutrient intake (vitamins A, C, K, folate, potassium, and beta-carotene reported on 3-day food records. F/V scale construct validity and internal consistency were assessed and compared to: 1) the original scale validation in Chinese women, 2) WEL scale, and 3) to micronutrient intake from 3-day food records. Total scale scores differed between African American women (μ=1.87+/-0.87) and Chinese (μ=0.41). In a Chinese population, F/V scale factored into two subscales; the F/V factored into one subscale in African American women. Construct validity was supported with correlation between the F/V scale and the eating efficacy WEL subscale (r 2 =-0.336, p=0.000). There was not a significant correlation between dietary consumption of micronutrients representative of fruit and vegetable intake and the F/V scale. The F/V scale developed for Chinese populations can be reliably used with African American women. Copyright © 2017 Elsevier Ltd. All rights reserved.

Veggie Van Pilot Study: Impact of a Mobile Produce Market for Underserved Communities on Fruit and Vegetable Access and Intake

PubMed Central

Leone, Lucia A.; Haynes-Maslow, Lindsey; Ammerman, Alice S.

2016-01-01

We conducted a pilot evaluation of the Veggie Van, a mobile produce market that brings weekly boxes of reduced-cost locally grown fruits and vegetables (F&V) to lower-income communities and offers cooking and nutrition education to customers. We conducted surveys just prior to starting Veggie Van at each of 3 sites and again at 2–3 months. F&V intake was measured with a 2-question item and a 10-item food frequency questionnaire (FFQ) in a subset of participants. At baseline, average servings/day of F&V was 4.9 (SD = 2.6, n = 60). At follow-up, individuals who reported shopping at Veggie Van frequently (n = 32) increased their F&V consumption by 0.41 servings/day compared with a decrease of −1.19 for those who rarely/never used Veggie Van (n = 27), a total difference of 1.6 servings/day (P = .01). There were no statistically significant differences in F&V consumption between groups based on the FFQ measure. Frequent shoppers reported additional health improvements and increases in their ability to buy enough F&V. We conclude that offering weekly boxes of affordable F&V paired with education in underserved communities may improve F&V consumption for frequent program users. PMID:28529669

Veggie Van Pilot Study: Impact of a Mobile Produce Market for Underserved Communities on Fruit and Vegetable Access and Intake.

PubMed

Leone, Lucia A; Haynes-Maslow, Lindsey; Ammerman, Alice S

2017-01-01

We conducted a pilot evaluation of the Veggie Van, a mobile produce market that brings weekly boxes of reduced-cost locally grown fruits and vegetables (F&V) to lower-income communities and offers cooking and nutrition education to customers. We conducted surveys just prior to starting Veggie Van at each of 3 sites and again at 2-3 months. F&V intake was measured with a 2-question item and a 10-item food frequency questionnaire (FFQ) in a subset of participants. At baseline, average servings/day of F&V was 4.9 (SD = 2.6, n = 60). At follow-up, individuals who reported shopping at Veggie Van frequently (n = 32) increased their F&V consumption by 0.41 servings/day compared with a decrease of -1.19 for those who rarely/never used Veggie Van (n = 27), a total difference of 1.6 servings/day (P = .01). There were no statistically significant differences in F&V consumption between groups based on the FFQ measure. Frequent shoppers reported additional health improvements and increases in their ability to buy enough F&V. We conclude that offering weekly boxes of affordable F&V paired with education in underserved communities may improve F&V consumption for frequent program users.

Insulated Foamy Viral Vectors

PubMed Central

Browning, Diana L.; Collins, Casey P.; Hocum, Jonah D.; Leap, David J.; Rae, Dustin T.; Trobridge, Grant D.

2016-01-01

Retroviral vector-mediated gene therapy is promising, but genotoxicity has limited its use in the clinic. Genotoxicity is highly dependent on the retroviral vector used, and foamy viral (FV) vectors appear relatively safe. However, internal promoters may still potentially activate nearby genes. We developed insulated FV vectors, using four previously described insulators: a version of the well-studied chicken hypersensitivity site 4 insulator (650cHS4), two synthetic CCCTC-binding factor (CTCF)-based insulators, and an insulator based on the CCAAT box-binding transcription factor/nuclear factor I (7xCTF/NF1). We directly compared these insulators for enhancer-blocking activity, effect on FV vector titer, and fidelity of transfer to both proviral long terminal repeats. The synthetic CTCF-based insulators had the strongest insulating activity, but reduced titers significantly. The 7xCTF/NF1 insulator did not reduce titers but had weak insulating activity. The 650cHS4-insulated FV vector was identified as the overall most promising vector. Uninsulated and 650cHS4-insulated FV vectors were both significantly less genotoxic than gammaretroviral vectors. Integration sites were evaluated in cord blood CD34+ cells and the 650cHS4-insulated FV vector had fewer hotspots compared with an uninsulated FV vector. These data suggest that insulated FV vectors are promising for hematopoietic stem cell gene therapy. PMID:26715244

Examination of the Relationship between In-Store Environmental Factors and Fruit and Vegetable Purchasing among Hispanics

PubMed Central

Sanchez-Flack, Jennifer; Pickrel, Julie L.; Belch, George; Lin, Shih-Fan; Anderson, Cheryl A. M.; Martinez, Maria Elena; Arredondo, Elva M.; Ayala, Guadalupe X.

2017-01-01

Retail food environments have received attention for their influence on dietary behaviors and for their nutrition intervention potential. To improve diet-related behaviors, such as fruit and vegetable (FV) purchasing, it is important to examine its relationship with in-store environmental characteristics. This study used baseline data from the “El Valor de Nuestra Salud” study to examine how in-store environmental characteristics, such as product availability, placement and promotion, were associated with FV purchasing among Hispanic customers in San Diego County. Mixed linear regression models indicated that greater availability of fresh FVs was associated with a $0.36 increase in FV purchasing (p = 0.01). Placement variables, specifically each additional square foot of display space dedicated to FVs (p = 0.01) and each additional fresh FV display (p = 0.01), were associated with a $0.02 increase and $0.29 decrease, respectively, in FV purchasing. Introducing FV promotions in the final model was not related to FV purchasing. Exploratory analyses indicated that men reported spending $3.69 fewer dollars on FVs compared to women, controlling for covariates (p = 0.02). These results can help inform interventions targeting in-store environmental characteristics to encourage FV purchasing among Hispanics. PMID:29077075

Documented family violence and risk of suicide attempt among U.S. Army soldiers.

PubMed

Ursano, Robert J; Stein, Murray B; Herberman Mash, Holly B; Naifeh, James A; Fullerton, Carol S; Zaslavsky, Alan M; Ng, Tsz Hin Hinz; Aliaga, Pablo A; Wynn, Gary H; Dinh, Hieu M; McCarroll, James E; Sampson, Nancy A; Kao, Tzu-Cheg; Schoenbaum, Michael; Heeringa, Steven G; Kessler, Ronald C

2018-04-01

Suicide attempt (SA) rates in the U.S. Army increased substantially during the wars in Afghanistan and Iraq. This study examined associations of family violence (FV) history with SA risk among soldiers. Using administrative data from the Army Study to Assess Risk and Resilience in Servicemembers (Army STARRS), we identified person-month records of active duty, Regular Army, enlisted soldiers with medically documented SAs from 2004 to 2009 (n = 9650) and a sample of control person-months (n = 153,528). Logistic regression analyses examined associations of FV with SA, adjusting for socio-demographics, service-related characteristics, and prior mental health diagnosis. Odds of SA were higher in soldiers with a FV history and increased as the number of FV events increased. Soldiers experiencing past-month FV were almost five times as likely to attempt suicide as those with no FV history. Odds of SA were elevated for both perpetrators and those who were exclusively victims. Male perpetrators had higher odds of SA than male victims, whereas female perpetrators and female victims did not differ in SA risk. A discrete-time hazard function indicated that SA risk was highest in the initial months following the first FV event. FV is an important consideration in understanding risk of SA among soldiers. Published by Elsevier B.V.

State farm-to-school laws influence the availability of fruits and vegetables in school lunches at US public elementary schools.

PubMed

Nicholson, Lisa; Turner, Lindsey; Schneider, Linda; Chriqui, Jamie; Chaloupka, Frank

2014-05-01

State laws and farm-to-school programs (FTSPs) have the potential to increase fruit and vegetable (FV) availability in school meals. This study examined whether FV were more available in public elementary school lunches in states with a law requiring/encouraging FTSPs or with a locally grown-related law, and whether the relationship between state laws and FV availability could be explained by schools opting for FTSPs. A pooled, cross-sectional analysis linked a nationally representative sample of public elementary schools with state laws. A series of multivariate logistic regressions, controlling for school-level demographics were performed according to mediation analysis procedures for dichotomous outcomes. Roughly 50% of schools reported FV availability in school lunches on most days of the week. Schools with the highest FV availability (70.6%) were in states with laws and schools with FTSPs. State laws requiring/encouraging FTSPs were significantly associated with increased FV availability in schools and a significant percentage (13%) of this relationship was mediated by schools having FTSPs. Because state farm-to-school laws are associated with significantly higher FV availability in schools-through FTSPs, as well as independently-enacting more state legislation may facilitate increased FTSP participation by schools and increased FV availability in school meals. © 2014, American School Health Association.

Collider signals of maximal flavor violation: Same-sign leptons from same-sign top quarks at the Fermilab Tevatron

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

Bar-Shalom, Shaouly; Department of Physics and Astronomy, University of California, Irvine, California 92697; Rajaraman, Arvind

2008-08-01

In models of maximal flavor violation (MxFV) suggested in [S. Bar-Shalom and A. Rajaraman, Phys. Rev. D 77, 095011 (2008).] there is at least one new scalar {phi}{sub FV} which couples to the quarks via {phi}{sub FV}q{sub i}q{sub j}{proportional_to}{xi}{sub ij} where {xi}{sub i3},{xi}{sub 3i}{approx}V{sub tb} for i=1, 2 and {xi}{sub 33}{approx}V{sub td} and V is the CKM matrix. In this article, we explore the potential phenomenological implications of MxFV for collider experiments. We study MxFV signals of same-sign leptons from same-sign top-quark pair production at the Tevatron and at the LHC. We show that the current Tevatron data set hasmore » strong sensitivity to this signature, for which there are no current limits. For example, if m{sub {phi}{sub FV}}{approx}200 GeV and the MxFV coupling {xi} has a natural value of {approx}1, we expect {approx}12 MxFV events to survive a selection requiring a pair of same-sign leptons, a tagged b-jet and missing transverse energy, over a background of approximately 4-5 events.« less

Are plasma homocysteine concentrations in Brazilian adolescents influenced by the intake of the main food sources of natural folate?

PubMed

Bigio, Roberta Schein; Verly, Eliseu; de Castro, Michelle Alessandra; Cesar, Chester Luis Galvão; Fisberg, Regina Mara; Marchioni, Dirce Maria Lobo

2013-01-01

Folate, a B vitamin, has been associated with a reduced concentration of plasma homocysteine (phcy), a marker of cardiovascular disease. The contribution of fruits and vegetables (FV) and other natural folate-rich foods to folate intake and folate status in Brazilian adolescents has hardly been determined. To investigate the intake of FV and beans and its association with the concentration of phcy in adolescents. This was a cross-sectional population-based study with a complex sample survey, with 198 adolescents who completed two 24-hour dietary recalls, a food frequency questionnaire, and a fasting blood draw. Usual dietary intake estimates were derived applying the Multiple Source Method. Three different generalized linear models with a gamma distribution were developed for each sex to evaluate the relationship between phcy and tertiles of FV intake as well as to evaluate the relationship between phcy and tertiles of FV and bean intake. No association was found between phcy concentration and FV intake or between phcy and FV and beans. Serum folate and female sex were inversely related to phcy. Phcy was not related to FV or FV and beans; this may be attributable to a low intake of these food groups. Copyright © 2013 S. Karger AG, Basel.

Histone H3 lysine 9 methyltransferase FvDim5 regulates fungal development, pathogenicity and osmotic stress responses in Fusarium verticillioides.

PubMed

Gu, Qin; Ji, Tiantian; Sun, Xiao; Huang, Hai; Zhang, Hao; Lu, Xi; Wu, Liming; Huo, Rong; Wu, Huijun; Gao, Xuewen

2017-10-16

Histone methylation plays important biological roles in eukaryotic cells. Methylation of lysine 9 at histone H3 (H3K9me) is critical for regulating chromatin structure and gene transcription. Dim5 is a lysine histone methyltransferase (KHMTase) enzyme, which is responsible for the methylation of H3K9 in eukaryotes. In the current study, we identified a single ortholog of Neurospora crassa Dim5 in Fusarium verticillioides. In this study, we report that FvDim5 regulates the trimethylation of H3K9 (H3K9me3). The FvDIM5 deletion mutant (ΔFvDim5) showed significant defects in conidiation, perithecium production and fungal virulence. Unexpectedly, we found that deletion of FvDIM5 resulted in increased tolerance to osmotic stresses and upregulated FvHog1 phosphorylation. These results indicate the importance of FvDim5 for the regulation of fungal development, pathogenicity and osmotic stress responses in F. verticillioides. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Vegetable and fruit juice enhances antioxidant capacity and regulates antioxidant gene expression in rat liver, brain and colon

PubMed Central

Yuan, Linhong; Liu, Jinmeng; Zhen, Jie; Xu, Yao; Chen, Shuying; Halm-Lutterodt, Nicholas Van; Xiao, Rong

2017-01-01

Abstract To explore the effect of fruit and vegetable (FV) juice on biomarkers of oxidative damage and antioxidant gene expression in rats, 36 adult male Wistar rats were randomly divided into control, low FV juice dosage or high FV juice dosage treatment groups. The rats were given freshly extracted FV juice or the same volume of saline water daily for five weeks. After intervention, serum and tissues specimens were collected for biomarker and gene expression measurement. FV juice intervention increased total antioxidant capacity, glutathione, vitamin C, β-carotene, total polyphenols, flavonoids levels andglutathione peroxidaseenzyme activity in rat serum or tissues (p < 0.05). FV juice intervention caused reduction of malondialdehyde levels in rat liver (p < 0.05) and significantly modulated transcript levels of glutamate cysteine ligase catalytic subunit (GCLC) and NAD(P)H:quinone oxidoreductase l (NQO1)in rat liver and brain (p < 0.05). The results underline the potential of FV juice to improve the antioxidant capacity and to prevent the oxidative damage in liver, brain and colon. PMID:28323302

The effectiveness of asking behaviors among 9-11 year-old children in increasing home availability and children's intake of fruit and vegetables: Results from the Squire's Quest II self-regulation game intervention

USDA-ARS?s Scientific Manuscript database

Home environment has an important influence on children's fruit and vegetable (FV) consumption, but children may in turn also impact their home FV environment, e.g. by asking for FV. The Squire's Quest II serious game intervention aimed to increase asking behaviors to improve home FV availability an...

[Relationship between fruit and vegetable gardening and health-related factors: male community gardeners aged 50-74 years living in a suburban area of Japan].

PubMed

Machida, Daisuke; Yoshida, Tohru

2017-01-01

Objectives　The aims of the study were as follows: 1) to investigate the relationship between community fruit and vegetable (FV) gardening and perceived changes in health-related factors by utilizing community gardens and 2) to determine the relationship of community FV gardening and other types of gardening on health-related factors among men aged 50-74 years living in a suburban area of Japan.Methods　In this cross-sectional study, we targeted men aged 50-74 years living in a city in Gunma Prefecture. A survey solicited demographic characteristics, FV gardening information, and health-related factors [BMI, self-rated health status, FV intake, physical activity (PA), and perceived neighborhood social cohesion (PNSC)]. The participants were divided into three groups: community gardeners, other types of gardeners, and non-gardeners. Items related to community gardening and perceived changes in health-related factors were presented only to community gardeners. The relationship between community gardening and perceived changes in health-related factors were analyzed by computing correlation coefficients. The relationships between FV gardening and specific health-related factors were analyzed by logistic regression modeling.Results　Significant positive correlations were observed between community FV gardening (the frequency of community gardening, the product of community gardening time and frequency of community gardening) and perceived changes in health-related factors (frequency of FV intake, amount of FV intake, and PA). The logistic regression models showed that 1) the number of participants with ≥23 METs h/week of PA was significantly greater among community gardeners than among non-gardeners; 2) the number of participants whose frequency of total vegetable intake, total vegetable intake (excluding juice), and total FV intake (excluding juice) was ≥5 times/day was significantly greater among other types of gardeners than non-gardeners; 3) participants with scores ≥ the median of PNSC were significantly greater among other types of gardeners than non-gardeners; and 4) participants who spent ≥4 hours/day sitting were significantly fewer among other types of gardeners than non-gardeners.Conclusion　Higher frequency of community gardening appears to induce greater perceived positive changes on FV intake and PA. It was indicated that FV gardening in community gardens contributes to increased PA, whereas other types of FV gardening contribute to increased FV intake frequency and decreased sitting time. In the future, higher-quality studies-for example, intervention studies using more rigorous measurements-will be necessary.

Chaperone-Assisted Soluble Expression of a Humanized Anti-EGFR ScFv Antibody in E. Coli

PubMed Central

Veisi, Kamal; Farajnia, Safar; Zarghami, Nosratollah; Khoram Khorshid, Hamid Reza; Samadi, Nasser; Ahdi Khosroshahi, Shiva; Zarei Jaliani, Hossein

2015-01-01

Purpose: Formation of inclusion bodies is a considerable obstacle threatening the advantages of E. coli expression system to serve as the most common and easiest system in recombinant protein production. To solve this problem, several strategies have been proposed among which application of molecular chaperones is of remarkable consideration. The aim of this study was to evaluate the effects of molecular chaperones on soluble expression of aggregation-prone humanized single chain antibody. Methods: To increase the solubility of a humanized single chain antibody (hscFv), different chaperone plasmids including PG-tf2 (GroES- GroEL- tig), ptf16 (tig) and pGro7 (GroES- GroEL) were co-expressed in BL21 cells containing pET-22b- hscFv construct. The solubility of recombinant hscFv was analyzed by SDS-PAGE. After purification of soluble hscFv by Ni-NTA column, the biological activity and cytotoxicity of the recombinant protein were tested by ELISA and MTT assay, respectively. Results: SDS-PAGE analysis of the hscFv revealed that chaperone utility remarkably increased (up to 50%) the solubility of the protein. ELISA test and MTT assay analyses also confirmed the biological activity of the gained hscFv in reaction with A431 cells (OD value: 2.6) and inhibition of their proliferation, respectively. Conclusion: The results of this study revealed that co-expression of chaperones with hscFv leads to remarkable increase in the solubility of the recombinant hscFv, which could be of great consideration for large scale production of recombinant single chain antibodies. PMID:26793607

Recombinant anti-podoplanin (NZ-1) immunotoxin for the treatment of malignant brain tumors

PubMed Central

Chandramohan, Vidyalakshmi; Bao, Xuhui; Kaneko, Mika Kato; Kato, Yukinari; Keir, Stephen T.; Szafranski, Scott E.; Kuan, Chien-Tsun; Pastan, Ira H.; Bigner, Darell D.

2013-01-01

Current study demonstrates the glioma tumor antigen podoplanin to be present at very high levels (>90%) in both glioblastoma (D2159MG, D08-0308MG, and D08-0493MG) and medulloblastoma (D283MED, D425MED, and DAOY) xenografts and cell line. We constructed a novel recombinant single-chain antibody variable region fragment (scFv), NZ-1, specific for podoplanin from the NZ-1 hybridoma. NZ-1-scFv was then fused to Pseudomonas exotoxin A, carrying a C-terminal KDEL peptide (NZ-1-PE38KDEL). The immunotoxin was further stabilized by a disulfide (ds) bond between the heavy-chain and light-chain variable regions as the construct NZ-1-(scdsFv)-PE38KDEL. NZ-1-(scdsFv)-PE38KDEL exhibited significant reactivity to glioblastoma and medulloblastoma cells. The affinity of NZ-1-(scdsFv), NZ-1-(scdsFv)-PE38KDEL and NZ-1 antibody, for podoplanin peptide was 2.1×10−8 M, 8.0×10−8 M, and 3.9×10−10 M, respectively. In a protein stability assay, NZ-1-(scdsFv)-PE38KDEL retained 33-98% of its activity while that of NZ-1-PE38KDEL declined to 13% of its initial levels after incubation at 37°C for 3 days. In vitro cytotoxicity of the NZ-1-(scdsFv)-PE38KDEL was measured in cells isolated from glioblastoma xenografts, D2159MG, D08-0308MG, D08-0493MG, and in the medulloblastoma D283MED, D425MED, and DOAY xenografts and cell line. The NZ-1-(scdsFv)-PE38KDEL immunotoxin was highly cytotoxic, with an IC50 in the range of 1.6–29 ng/mL. Significantly, NZ-1-(scdsFv)-PE38KDEL demonstrated tumor-growth delay, averaging 24 days (P<0.001) and 21 days (P<0.001) in D2159MG and D283MED in vivo tumor models, respectively. Crucially, in the D425MED intracranial tumor model, NZ-1-(scdsFv)-PE38KDEL caused a 41% increase in survival (P≤0.001). In preclinical studies, NZ-1-(scdsFv)-PE38KDEL exhibited significant potential as a targeting agent for malignant brain tumors. PMID:23115013

Recombinant anti-podoplanin (NZ-1) immunotoxin for the treatment of malignant brain tumors.

PubMed

Chandramohan, Vidyalakshmi; Bao, Xuhui; Kato Kaneko, Mika; Kato, Yukinari; Keir, Stephen T; Szafranski, Scott E; Kuan, Chien-Tsun; Pastan, Ira H; Bigner, Darell D

2013-05-15

Our study demonstrates the glioma tumor antigen podoplanin to be present at very high levels (>90%) in both glioblastoma (D2159MG, D08-0308MG and D08-0493MG) and medulloblastoma (D283MED, D425MED and DAOY) xenografts and cell line. We constructed a novel recombinant single-chain antibody variable region fragment (scFv), NZ-1, specific for podoplanin from the NZ-1 hybridoma. NZ-1-scFv was then fused to Pseudomonas exotoxin A, carrying a C-terminal KDEL peptide (NZ-1-PE38KDEL). The immunotoxin (IT) was further stabilized by a disulfide (ds) bond between the heavy-chain and light-chain variable regions as the construct NZ-1-(scdsFv)-PE38KDEL. NZ-1-(scdsFv)-PE38KDEL exhibited significant reactivity to glioblastoma and medulloblastoma cells. The affinity of NZ-1-(scdsFv), NZ-1-(scdsFv)-PE38KDEL and NZ-1 antibody for podoplanin peptide was 2.1 × 10(-8) M, 8.0 × 10(-8) M and 3.9 × 10(-10) M, respectively. In a protein stability assay, NZ-1-(scdsFv)-PE38KDEL retained 33-98% of its activity, whereas that of NZ-1-PE38KDEL declined to 13% of its initial levels after incubation at 37°C for 3 days. In vitro cytotoxicity of the NZ-1-(scdsFv)-PE38KDEL was measured in cells isolated from glioblastoma xenografts, D2159MG, D08-0308MG and D08-0493MG, and in the medulloblastoma D283MED, D425MED and DOAY xenografts and cell line. The NZ-1-(scdsFv)-PE38KDEL IT was highly cytotoxic, with an 50% inhibitory concentration in the range of 1.6-29 ng/ml. Significantly, NZ-1-(scdsFv)-PE38KDEL demonstrated tumor growth delay, averaging 24 days (p < 0.001) and 21 days (p < 0.001) in D2159MG and D283MED in vivo tumor models, respectively. Crucially, in the D425MED intracranial tumor model, NZ-1-(scdsFv)-PE38KDEL caused a 41% increase in survival (p ≤ 0.001). In preclinical studies, NZ-1-(scdsFv)-PE38KDEL exhibited significant potential as a targeting agent for malignant brain tumors. Copyright © 2012 UICC.

Cluster randomized controlled trial of a mobile market intervention to increase fruit and vegetable intake among adults in lower-income communities in North Carolina.

PubMed

Leone, Lucia A; Tripicchio, Gina L; Haynes-Maslow, Lindsey; McGuirt, Jared; Grady Smith, Jacqueline S; Armstrong-Brown, Janelle; Gizlice, Ziya; Ammerman, Alice

2018-01-05

Poorer diets and subsequent higher rates of chronic disease among lower-income individuals may be partially attributed to reduced access to fresh fruits and vegetables (F&V) and other healthy foods. Mobile markets are an increasingly popular method for providing access to F&V in underserved communities, but evaluation efforts are limited. The purpose of this study was to determine the impact of Veggie Van (VV), a mobile produce market, on F&V intake in lower-income communities using a group randomized controlled trial. VV is a mobile produce market that sells reduced-cost locally grown produce and offers nutrition and cooking education. We recruited 12 sites in lower-income communities in North Carolina (USA) to host VV, randomizing them to receive VV immediately (intervention) or after the 6-month study period (delayed intervention control). Participants at each site completed baseline and follow-up surveys including F&V intake, perceived access to fresh F&V and self-efficacy for purchasing, preparing and eating F&V. We used multiple linear regression to calculate adjusted differences in outcomes while controlling for baseline values, education and clustering within site. Among 142 participants who completed the follow-up, baseline F&V intake was 3.48 cups/day for control and 3.33 for intervention. At follow-up, adjusted change in F&V consumption was 0.95 cups/day greater for intervention participants (p = 0.005), but was attenuated to 0.51 cups per day (p = 0.11) after removing extreme values. VV customers increased their F&V consumption by 0.41 cups/day (n = 30) compared to a 0.25 cups/day decrease for 111 non-customers (p = 0.04). Intervention participants did not show significant improvements in perceived access to fresh F&V, but increased their self-efficacy for working more F&V into snacks (p = 0.02), making up a vegetable dish with what they had on hand (p = 0.03), and cooking vegetables in a way that is appealing to their family (p = 0.048). Mobile markets may help improve F&V intake in lower-income communities. Clinicaltrials.gov ID# NCT03026608 retrospectively registered January 2, 2017.

Systemic p53 gene therapy of cancer with immunolipoplexes targeted by anti-transferrin receptor scFv.

PubMed Central

Xu, L.; Tang, W. H.; Huang, C. C.; Alexander, W.; Xiang, L. M.; Pirollo, K. F.; Rait, A.; Chang, E. H.

2001-01-01

BACKGROUND: A long-standing goal in genetic therapy for cancer is a systemic gene delivery system that selectively targets tumor cells, including metastases. Here we describe a novel cationic immunolipoplex system that shows high in vivo gene transfer efficiency and anti- tumor efficacy when used for systemic p53 gene therapy of cancer. MATERIALS AND METHODS: A cationic immunolipoplex incorporating a biosynthetically lipid-tagged, anti-transferrin receptor single-chain antibody (TfRscFv), was designed to target tumor cells both in vitro and in vivo. A human breast cancer metastasis model was employed to evaluate the in vivo efficacy of systemically administered, TfRscFv-immunolipoplex-mediated, p53 gene therapy in combination with docetaxel. RESULTS: The TfRscFv-targeting cationic immunolipoplex had a size of 60-100 nm, showed enhanced tumor cell binding, and improved targeted gene delivery and transfection efficiencies, both in vitro and in vivo. The p53 tumor suppressor gene was not only systemically delivered by the immunolipoplex to human tumor xenografts in nude mice but also functionally expressed. In the nude mouse breast cancer metastasis model, the combination of the p53 gene delivered by the systemic administration of the TfRscFv-immunolipoplex and docetaxel resulted in significantly improved efficacy with prolonged survival. CONCLUSIONS: This is the first report using scFv-targeting immunolipoplexes for systemic gene therapy. The TfRscFv has a number of advantages over the transferrin (Tf) molecule itself: (1) scFv has a much smaller size than Tf producing a smaller immunolipoplex giving better penetration into solid tumors; (2) unlike Tf, the scFv is a recombinant protein, not a blood product; (3) large scale production and strict quality control of the recombinant scFv, as well as scFv-immunolipoplex, are feasible. The sensitization of tumors to chemotherapy by this tumor-targeted and efficient p53 gene delivery method could lower the effective dose of the drug, correspondingly lessening the severe side effects, while decreasing the possibility of recurrence. Moreover, this approach is applicable to both primary and recurrent tumors, and more significantly, metastatic disease. The TfRscFv-targeting of cationic immunolipoplexes is a promising method of tumor targeted gene delivery that can be used for systemic gene therapy of cancer with the potential to critically impact the clinical management of cancer. PMID:11713371

A multi-level intervention in subsidized housing sites to increase fruit and vegetable access and intake: Rationale, design and methods of the 'Live Well, Viva Bien' cluster randomized trial.

PubMed

Gans, Kim M; Gorham, Gemma; Risica, Patricia M; Dulin-Keita, Akilah; Dionne, Laura; Gao, Tina; Peters, Sarah; Principato, Ludovica

2016-06-28

Adequate fruit and vegetable (F&V) intake is important for disease prevention. Yet, most Americans, especially low-income and racial/ethnic minorities, do not eat adequate amounts. These disparities are partly attributable to food environments in low-income neighborhoods where residents often have limited access to affordable, healthful food and easy access to inexpensive, unhealthful foods. Increasing access to affordable healthful food in underserved neighborhoods through mobile markets is a promising, year-round strategy for improving dietary behaviors and reducing F&V intake disparities. However, to date, there have been no randomized controlled trials studying their effectiveness. The objective of the 'Live Well, Viva Bien' (LWVB) cluster randomized controlled trial is to evaluate the efficacy of a multicomponent mobile market intervention at increasing F&V intake among residents of subsidized housing complexes. One housing complex served as a pilot site for the intervention group and the remaining 14 demographically-matched sites were randomized into either the intervention or control group. The intervention group received bimonthly, discount, mobile, fresh F&V markets in conjunction with a nutrition education intervention (two F&V campaigns, newsletters, DVDs and cooking demonstrations) for 12 months. The control group received physical activity and stress reduction interventions. Outcome measures include F&V intake (measured by two validated F&V screeners at baseline, six-month and twelve-months) along with potential psychosocial mediating variables. Extensive quantitative and qualitative process evaluation was also conducted throughout the study. Modifying neighborhood food environments in ways that increase access to affordable, healthful food is a promising strategy for improving dietary behaviors among low-income, racial and ethnic minority groups at increased risk for obesity and other food-related chronic diseases. Discount, mobile F&V markets address all the major barriers to eating more F&V (high cost, poor quality, limited access and limited time to shop and cook) and provide a year-round solution to limited access to healthful food in low-income neighborhoods. LWVB is the first randomized controlled trial evaluating the effectiveness of mobile markets at increasing F&V intake. If proven efficacious at increasing F&V consumption, LWVB could be disseminated widely to neighborhoods that have low access to fresh F&V. Clinicatrials.gov registration number: NCT02669472 First Received: January 19, 2016.

The intervening removable affinity tag (iRAT) production system facilitates Fv antibody fragment‐mediated crystallography

PubMed Central

Nomura, Yayoi; Sato, Yumi; Suno, Ryoji; Horita, Shoichiro

2016-01-01

Abstract Fv antibody fragments have been used as co‐crystallization partners in structural biology, particularly in membrane protein crystallography. However, there are inherent technical issues associated with the large‐scale production of soluble, functional Fv fragments through conventional methods in various expression systems. To circumvent these problems, we developed a new method, in which a single synthetic polyprotein consisting of a variable light (VL) domain, an intervening removable affinity tag (iRAT), and a variable heavy (VH) domain is expressed by a Gram‐positive bacterial secretion system. This method ensures stoichiometric expression of VL and VH from the monocistronic construct followed by proper folding and assembly of the two variable domains. The iRAT segment can be removed by a site‐specific protease during the purification process to yield tag‐free Fv fragments suitable for crystallization trials. In vitro refolding step is not required to obtain correctly folded Fv fragments. As a proof of concept, we tested the iRAT‐based production of multiple Fv fragments, including a crystallization chaperone for a mammalian membrane protein as well as FDA‐approved therapeutic antibodies. The resulting Fv fragments were functionally active and crystallized in complex with the target proteins. The iRAT system is a reliable, rapid and broadly applicable means of producing milligram quantities of Fv fragments for structural and biochemical studies. PMID:27595817

A farmers' market at a federally qualified health center improves fruit and vegetable intake among low-income diabetics.

PubMed

Freedman, Darcy A; Choi, Seul Ki; Hurley, Thomas; Anadu, Edith; Hébert, James R

2013-05-01

A 22-week federally qualified health center (FQHC)-based farmers' market (FM) and personal financial incentive intervention designed to improve access to and consumption of fruits and vegetables (FVs) among low-income diabetics in rural South Carolina was evaluated. A mixed methods, one-group, repeated-measures design was used. Data were collected in 2011 before (May/June), during (August), and after (November) the intervention with 41 diabetes patients from the FQHC. FV consumption was assessed using a validated National Cancer Institute FV screener modified to include FV sold at the FM. Sales receipts were recorded for all FM transactions. A mixed-model, repeated measures analysis of variance was used to assess intervention effects on FV consumption. Predictors of changes in FV consumption were examined using logistic regression. A marginally significant (p=0.07) average increase of 1.6 servings of total FV consumption per day occurred. The odds of achieving significant improvements in FV consumption increased for diabetics using financial incentives for payment at the FM (OR: 38.8, 95% CI: 3.4-449.6) and for those frequenting the FM more often (OR: 2.1, 95% CI: 1.1-4.0). Results reveal a dose-response relationship between the intervention and FV improvements and emphasize the importance of addressing economic barriers to food access. Copyright © 2013 Elsevier Inc. All rights reserved.

The development and effectiveness of an ecological momentary intervention to increase daily fruit and vegetable consumption in low-consuming young adults.

PubMed

Brookie, Kate L; Mainvil, Louise A; Carr, Anitra C; Vissers, Margreet C M; Conner, Tamlin S

2017-01-01

To develop and test the effectiveness of a mobile-phone based ecological momentary intervention (EMI) to increase fruit and vegetable (FV) consumption in low-consuming young adults. A two-week randomised controlled trial of low-FV consuming young adults ages 18-25 years (n = 171) compared three conditions: ecological momentary intervention (EMI), fruit and vegetable intervention (FVI), and a diet-as-usual control (ANZCTRN12615000183583). Participants in the EMI condition were sent two targeted text messages a day for 13 days and were asked to increase daily FV consumption to at least five servings. These messages were designed, using the Health Action Process Approach, to address salient beliefs identified as effective in a preliminary focus group investigation. Participants in the FVI condition were provided two servings of FV a day (carrots, kiwifruit or oranges, and apples) to eat in addition to their current diet. Control participants ate their normal diet. Participants reported their daily servings of FV each evening during the study using a smartphone-delivered survey. Blood samples testing plasma vitamin C and total carotenoids were taken pre- and post-intervention as an objective biomarker of FV intake. Participants in the EMI and FVI conditions reported higher daily servings of FV - approximately +1 serving per day more compared to control (EMI = 3.7 servings/day; FVI = 3.7 servings/day; Control = 2.8 servings/day) and approximately +1.2 servings compared to baseline. Increases in objective biomarkers for the experimental conditions supported the validity of self-reported FV consumption. Our results provide initial proof of concept that EMI strategies (with minor financial assistance) are as effective as giving FV in increasing FV consumption in educated, low-consuming young adults. Copyright © 2016 Elsevier Ltd. All rights reserved.

Simple, monolithic optical element for forward-viewing spectrally encoded endoscopy (Conference Presentation)

NASA Astrophysics Data System (ADS)

Do, Dukho; Kang, Dongkyun; Ikuta, Mitsuhiro; Tearney, Guillermo J.

2016-03-01

Spectrally encoded endoscopy (SEE) is a miniature endoscopic technology that can acquire images of internal organs through a hair-thin probe. While most previously described SEE probes have been side viewing, forward-view (FV)-SEE is advantageous in certain clinical applications as it provides more natural navigation of the probe and has the potential to provide a wider field of view. Prior implementations of FV-SEE used multiple optical elements that increase fabrication complexity and may diminish the robustness of the device. In this paper, we present a new design that uses a monolithic optical element to realize FV-SEE imaging. The optical element is specially designed spacer, fabricated from a 500-μm-glass rod that has a mirror surface on one side and a grating stamped on its distal end. The mirror surface is used to change the incident angle on the grating to diffract the shortest wavelength of the spectrum so that it is parallel to the optical axis. Rotating the SEE optics creates a circular FV-SEE image. Custom-designed software processes FV-SEE images into circular images, which are displayed in real-time. In order to demonstrate this new design, we have constructed the FV-SEE optical element using a 1379 lines/mm diffraction grating. When illuminated with a source with a spectral bandwidth of 420-820 nm, the FV-SEE optical element provides 678 resolvable points per line. The imaging performance of the FV-SEE device was tested by imaging a USAF resolution target. SEE images showed that this new approach generates high quality images in the forward field with a field of view of 58°. Results from this preliminary study demonstrate that we can realize FV-SEE imaging with simple, monolithic, miniature optical element. The characteristics of this FV-SEE configuration will facilitate the development of robust miniature endoscopes for a variety of medical imaging applications.

Effects of GSTM1/GSTT1 gene polymorphism and fruit & vegetable consumption on antioxidant biomarkers and cognitive function in the elderly: a community based cross-sectional study.

PubMed

Yuan, Linhong; Ma, Weiwei; Liu, Jinmeng; Meng, Liping; Liu, Jixia; Li, Shuang; Han, Jing; Liu, Quanri; Feng, Lingli; Wang, Chao; Xiao, Rong

2014-01-01

It was reported that Glutathione S-transferase (GST) gene polymorphism and fruit and vegetable (FV) intake were associated with body antioxidant capacity. The oxidative/anti-oxidative imbalance played an important role in the pathogenesis of AD. However, the association of GST genotype, dietary FV consumption with body antioxidant biomarkers and cognitive function in the elderly is not clear. The aim of the present study was to determine the association of GST genotype, and dietary FV intake, with antioxidant biomarkers and cognitive function in the elderly. Food frequency questionnaire was used to collect data of dietary FV intakes in 504 community dwelling elderly aged from 55 to 75 years old. GSTM1 and GSTT1 genotypes were determined by using multiple-PCR method. Plasma and erythrocyte antioxidant biomarkers were measured. Cognitive function was measured by using Montreal Cognitive Assessment. Statistical analysis was applied for exploring the association of GST genotype and FV intake with antioxidant biomarkers level and cognitive function in the elderly. Individual GSTM1 or GSTT1 gene deletion affects body antioxidant biomarkers levels, including erythrocyte GST activity, plasma total antioxidant capacity, and glutathione levels. GSTM1and/or GSTT1 gene deletion have no effects on cognitive function in the surveyed participants. The effect of GST genotype on antioxidant biomarkers are FV intake dependent. There is interaction of FV intake and GST genotype on cognitive function in the elderly. GST genotype or daily FV consumption impact body antioxidant biomarkers, but not cognitive function in the elderly. There were combined effects of GST genotype and FV consumption on cognitive function in the elderly population. Large scale perspective population study is required to explore the association of GST genetic polymorphism, FV consumption and antioxidant biomarkers and cognitive function in the elderly.

Adolescent fruit and vegetable intake: influence of family support and moderation by home availability of relationships with afrocentric values and taste preferences.

PubMed

Di Noia, Jennifer; Byrd-Bredbenner, Carol

2013-06-01

Economically disadvantaged African-American adolescents have fruit and vegetable (F/V) intakes that are less than optimal. To facilitate intervention planning to address low F/V intake in this population, an understanding of determinants of youths' intake is needed. The influence of determinants consistently supported by evidence (ie, home F/V availability, F/V taste preferences, and parental modeling/intake) and variables hypothesized to influence intake in the targeted population (ie, family support for F/V consumption and Afrocentric values) were examined. Participants were African-American adolescents recruited in 2011 through summer camps serving low-income youths (N=93). Youths completed a cross-sectional survey. Hierarchical logistic regression analysis was used to examine whether availability directly influenced (ie, explained variations in) intake and whether it moderated (ie, affected the direction and/or strength of) the relationships between the other hypothesized determinants and intake. The dependent variable was intake of five or more daily servings of F/V estimated with the Block 7-item food frequency questionnaire. Family support was directly related to intake (odds ratio=1.062; 95% CI 1.007 to 1.120; P=0.026). The relationships between F/V intake and taste preferences and Afrocentric values were moderated by (ie, differed based on) home F/V availability. When availability was high, taste preferences (odds ratio=1.081; 95% CI 1.007 to 1.161; P=0.032) and Afrocentric values (OR=2.504; 95% CI 1.303 to 4.811; P=0.006) had positive influences on intake. To enhance intervention effectiveness, more research is warranted on approaches for increasing home F/V availability and family support for F/V consumption in the targeted population. Copyright © 2013 Academy of Nutrition and Dietetics. Published by Elsevier Inc. All rights reserved.

Augmenting fruit and vegetable consumption by an online intervention: Psychological mechanisms.

PubMed

Keller, Jan; Motter, Susannah; Motter, Mirjam; Schwarzer, Ralf

2018-01-01

Fruit and vegetable (FV) intake was examined among men and women who participated in an online intervention. The psychological constructs involved were outcome expectancies, behavioral intention, planning, and self-efficacy. One purpose of the analyses was the evaluation of a self-efficacy treatment component. The other purpose of the analyses regarded the role of psychological mechanisms that might be responsible for individual differences in the process of behavior change. A two-arm online intervention with a standard and an enhanced intervention group focusing on FV planning was conducted to improve FV intake, followed up at two and four weeks. The intervention groups differed by the additional inclusion of a self-efficacy ingredient in the enhanced intervention. Linear mixed models examined the intervention effects, and a longitudinal structural equation model explored which psychological constructs were associated with changes in FV intake. Participants were N = 275 adults of whom n = 148 completed the four-week follow-up. Their age range was 18-81 years (M age  = 32.50, SD age  = 14.00). Analyses yielded an overall increase in self-reported FV intake. Moreover, a triple interaction between time, sex, and experimental groups on self-efficacy emerged, indicating that men, independent of treatment conditions, reported an increase in their confidence to improve FV intake, whereas women developed higher FV self-efficacy when being in the enhanced group instead of the standard group. Planning, self-efficacy, and intention mediated between outcome expectancies, and follow-up FV intake. Both intervention arms produced overall improvements in FV intake. The enhanced intervention resulted in a steeper increase in self-efficacy in women compared to men, and compared to the standard intervention. A psychological mechanism transpired that included a sequence leading from initial outcome expectancies via planning, self-efficacy, and intention towards FV intake. Copyright © 2017 Elsevier Ltd. All rights reserved.

Weight resilience and fruit and vegetable intake among African-American women in an obesogenic environment.

PubMed

Parisi, Sara M; Bodnar, Lisa M; Dubowitz, Tamara

2018-02-01

To investigate relationships between weight resilience (maintaining a normal weight in a food desert environment) and fruit and vegetable (F&V) intake, attitudes and barriers. Cross-sectional, in-person surveys collected May-December 2011, including self-reported data on F&V-related psychosocial factors, attitudes and barriers. Two 24 h dietary recalls were completed; weight and height were measured. Multivariable regression models estimated prevalence ratios (95 % CI). Two low-income, predominantly African-American food deserts in Pittsburgh, Pennsylvania, USA. Women aged 18-49 years (n 279) who were the primary food shopper in a household randomly selected for a parent study. Fifteen per cent were weight resilient, 30 % were overweight and 55 % were obese. Overall, 25 % reported eating ≥5 F&V servings/d. After adjustment for age, education, parity, employment, living alone, physical activity, per capita income and mean daily energy intake, women eating ≥5 F&V servings/d were 94 % more likely to be weight resilient compared with those eating <5 servings/d (1·94; 1·10, 3·43). Across BMI groups, self-efficacy regarding F&V consumption was high and few F&V barriers were reported. The most frequently reported barrier was concern about the cost of F&V (36 %). Of the attitudinal F&V-related factors, only concern about wasting food when serving F&V was associated with weight resilience in adjusted models (0·29; 0·09, 0·94). In a model predicting consuming ≥5 F&V servings/d, driving one's own car to the store was the only attitudinal F&V-related factor associated with consumption (1·50; 1·00, 2·24). In this population, weight resilience may be encouraged by improving access to affordable and convenient F&V options and providing education on ways to make them palatable to the entire household, rather than by shifting women's F&V perceptions, which are already positive.

Refolding of autodisplayed anti-NEF scFv through oxidation with glutathione for immunosensors.

PubMed

Bong, Ji-Hong; Song, Hyun-Woo; Kim, Tae-Hun; Kang, Min-Jung; Jose, Joachim; Pyun, Jae-Chul

2018-04-15

In this study, a single-domain antibody against negative regulatory factor (anti-NEF scFv) was autodisplayed on the outer membrane of Escherichia coli and used to detect NEF in an immunoassay based on fluorescence-activated cell sorting, enzyme-linked immunosorbent assay, and surface plasmon resonance biosensors. Next, the autodisplayed single-domain antibody was oxidized to form disulfide bonds by using glutathione, and the change in NEF-binding activity of anti-NEF scFv was analyzed by fluorescence-activated cell sorting-based immunoassay, chromogenic immunoassay, and surface plasmon resonance biosensor. For each type of immunoassays the anti-NEF scFv on the isolated outer membrane showed more NEF binding activity after the disulfide bond formation by glutathione. To determine the role of cysteines in anti-NEF scFv, three mutants were prepared, and the NEF binding activity of mutants was compared with that of wild-type anti-NEF scFv in a competitive immunoassay based on FACS. In these mutant studies, the refolding process of autodisplayed anti-NEF scFv by following oxidation via GSH/GSSG revealed that disulfide bonds formed and increased NEF binding activity. Copyright © 2017 Elsevier B.V. All rights reserved.

Perceived Influences on Farmers' Market Use among Urban, WIC-enrolled Women.

PubMed

Di Noia, Jennifer; Monica, Dorothy; Cullen, Karen Weber; Thompson, Debbe

2017-09-01

We identified perceived barriers and facilitators to purchasing fruits and vegetables (FV) at farmers' markets, FV shopping practices, and reactions to a planned online lesson to promote farmers' market use among urban, inner-city WIC-enrolled women. Thirteen focus groups were conducted with 3-5 participants each (N = 54). Common barriers were structural (transportation issues) and informational (not knowing the locations of markets). Improving access (by increasing the number of area markets, expanding market hours and locations, and increasing transportation options to markets) and raising awareness of the importance of eating healthfully were common facilitators. Information was sought on the locations of farmers who accept FV vouchers provided by WIC, FV sold at farmers' markets, reasons to eat locally grown FV, and FV food safety and preparation skills. Questions were raised about the accessibility of an online lesson; providing information via inperson seminars and handouts also was recommended. Although purchasing FV at supermarkets and corner stores and bodegas was common, concerns were expressed about the freshness, cost, quality, and variety of produce sold at these venues. Findings aid understanding of factors to consider in designing interventions to promote farmers' market use in this population.

Improved Brain Expression of Anti-Amyloid β scFv by Complexation of mRNA Including a Secretion Sequence with PEG-based Block Catiomer.

PubMed

Perche, Federico; Uchida, Satoshi; Akiba, Hiroki; Lin, Chin-Yu; Ikegami, Masaru; Dirisala, Anjaneyulu; Nakashima, Toshihiro; Itaka, Keiji; Tsumoto, Kohei; Kataoka, Kazunori

2017-01-01

The ever-increasing number of people living with Alzheimer's disease urges to develop more effective therapies. Despite considerable success, anti-Alzheimer immunotherapy still faces the challenge of intracerebral and intracellular delivery. This work introduces in situ production of anti-amyloid beta (Aβ) antibody after intracerebral injection of PEG-PAsp(DET)/mRNA polyplexes as a novel immunotherapy approach and a safer alternative compared to high systemic antibodies doses or administration of adenovirus encoding anti- Aβ antibodies. We used mRNA encoding three different Aβ-specific scFV with a secretion signal for passive immunotherapy. scFv contained a 6xHis-tag for immuno-detection. The secretion signal from IL2 (IL2ss) was added to allow extracellular engagement of senile plaques. Aβ affinity of scFv was measured by surface plasmon resonance. To allow intracellular delivery, scFv were administered as polyplexes formed with our smart copolymer polyethylene glycol-poly[N'-[N-(2-aminoethyl)-2-aminoethyl] aspartamide] [PEG-PAsp (DET)]. We evaluated scFv expression in cellulo by Western blot and ELISA, their ability to disaggregate amyloid aggregates by thioflavine T assay. Moreover, in vivo expression and therapeutic activity were evaluated in a murine amyloidosis model, by anti-6xHis-tag ELISA and anti- Aβ ELISA, respectively. The selected anti-amyloid beta scFv showed affinity towards Aβ and disaggregated Aβ fibers in vitro. Whereas both DNA and mRNA transfection led to scFV expression in cancer cells, only mRNA led to detectable scFv expression in primary neurons. In addition, the use of IL2ss increased by 3.4-fold scFv secretion by primary neurons over mRNA polyplexes devoid of secretion signal. In vivo, a 3 to 11- fold of intracranial scFv levels was measured for mRNA compared to DNA polyplexes and higher in vivo scFv levels were obtained with mRNA containing IL2ss over non-secreted mRNA. Intracranial injection of anti-Aβ mRNA polyplexes with IL2ss resulted in 40 % Aβ decrease in an acute amyloidosis model; with no decrease detected with control scFv mRNA nor DNA polyplexes. However, no Aβ decrease was detected in a more challenging transgenic model of Alzheimer's disease. Our results introduce a concerted approach not only for Alzheimer's disease treatment but also for immunotherapy against neurological diseases. The effectivity of our platform required the intracranial delivery of anti-Aβ scFv as mRNA not DNA, as mRNA with an IL2ss secretion sequence to favor engagement of Aβ in the amyloidosis model, complexation with a smart copolymer for efficient transfection of primary neurons and to achieve detectable mRNA expression in the brain during 48h. Amyloid burden decrease in an acute amyloidosis model was only achieved when these three factors (mRNA coding scFv, smart copolymer, IL2ss) were integrated into a single formulation. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

Screening of a ScFv Antibody With High Affinity for Application in Human IFN-γ Immunoassay

PubMed Central

Yang, Hang; Zhong, Yanfang; Wang, Juncheng; Zhang, Qinghong; Li, Xiulan; Ling, Sumei; Wang, Shihua; Wang, Rongzhi

2018-01-01

Interferon gamma (IFN-γ), a signal proinflammatory cytokine secreted by immune cell, and plays a critical role in the pathogenesis and progression of many diseases. It has been regarded as an important marker for determination of disease-specific immune responses. Therefore, it is urgent to develop a feasible and accurate method to detect IFN-γ in clinic real blood samples. Until now, the immunoassay based on singe chain variable fragment (scFv) antibody for human IFN-γ is still not reported. In the present study, an scFv antibody named scFv-A8 with high specificity was obtained by phage display and biopanning, with the affinity 2.6 × 109 L/mol. Maltose binding protein (MBP) was used to improve the solubility of scFv by inserting an linker DNA between scFv and MBP tag, and the resulted fusion protein (MBP-LK-scFv) has high solubility and antigen biding activity. The expressed and purified MBP-LK-scFv antibody was used to develop the indirect competitive enzyme-linked immunosorbent assay (ELISA) (ic-ELISA) for detection of human IFN-γ, and the result indicated that the linear range to detect IFN-γ was 6–60 pg/mL with IC50 of 25 pg/mL. The limit of detection was 2 pg/mL (1.3 fm), and the average recovery was 85.05%, further demonstrating that the detection method based on scFv has higher recovery and accuracy. Hence, the developed ic-ELISA can be used to detect IFN-γ in real samples, and it may be further provided a scientific basis for disease diagnosis. PMID:29563896

Biocompatible coupling of therapeutic fusion proteins to human erythrocytes

PubMed Central

Villa, Carlos H.; Pan, Daniel C.; Johnston, Ian H.; Greineder, Colin F.; Walsh, Landis R.; Hood, Elizabeth D.; Cines, Douglas B.; Poncz, Mortimer; Siegel, Don L.

2018-01-01

Carriage of drugs by red blood cells (RBCs) modulates pharmacokinetics, pharmacodynamics, and immunogenicity. However, optimal targets for attaching therapeutics to human RBCs and adverse effects have not been studied. We engineered nonhuman-primate single-chain antibody fragments (scFvs) directed to human RBCs and fused scFvs with human thrombomodulin (hTM) as a representative biotherapeutic cargo (hTM-scFv). Binding fusions to RBCs on band 3/glycophorin A (GPA; Wright b [Wrb] epitope) and RhCE (Rh17/Hr0 epitope) similarly endowed RBCs with hTM activity, but differed in their effects on RBC physiology. scFv and hTM-scFv targeted to band 3/GPA increased membrane rigidity and sensitized RBCs to hemolysis induced by mechanical stress, while reducing sensitivity to hypo-osmotic hemolysis. Similar properties were seen for other ligands bound to GPA and band 3 on human and murine RBCs. In contrast, binding of scFv or hTM-scFv to RhCE did not alter deformability or sensitivity to mechanical and osmotic stress at similar copy numbers bound per RBCs. Contrasting responses were also seen for immunoglobulin G antibodies against band 3, GPA, and RhCE. RBC-bound hTM-scFv generated activated protein C (APC) in the presence of thrombin, but RhCE-targeted hTM-scFv demonstrated greater APC generation per bound copy. Both Wrb- and RhCE-targeted fusion proteins inhibited fibrin deposition induced by tumor necrosis factor-α in an endothelialized microfluidic model using human whole blood. RhCE-bound hTM-scFv more effectively reduced platelet and leukocyte adhesion, whereas anti-Wrb scFv appeared to promote platelet adhesion. These data provide a translational framework for the development of engineered affinity ligands to safely couple therapeutics to human RBCs. PMID:29365311

Fruit and Vegetable Exposure in Children is Linked to the Selection of a Wider Variety of Healthy Foods at School

PubMed Central

Korinek, Elizabeth V.; Bartholomew, John B.; Jowers, Esbelle M.; Latimer, Lara A.

2013-01-01

Schools often offer healthy fruits and vegetables (FV) and healthy entrées. However, children may resist these efforts due to a lack of familiarity with the offerings. While numerous exposures with a food increase its liking, it may be that an exposure to a variety of FV at home leads to greater willingness to select other foods – even those that are unrelated to those eaten at home. As an initial test of this possibility, this study was designed to examine how self-reports of exposure and consumption of various FV were associated with the selection of FV and lunch entrées at school. Participants (N=59) were a convenience sample of elementary children. A median-split was used to place students into high and low exposure groups for self-reports of both exposure and consumption at home. The primary dependent variables were: self-reports of selecting FV at school; the children’s absolute and relative ratings of eight “healthier” lunch entrées; and self-reports of selecting these entrées. These entrées were recently added to the school menu and, therefore, tended to be less familiar to children. Food ratings were collected through taste exposures conducted at school. Results indicate that children who reported more frequent exposure to FV at home consumed a wider variety of FV at school and were more likely to report selecting “healthier” entrées at school lunch. These data suggest that exposure to and the consumption of a variety of FV may make children more willing to select a wider range of FV and other, healthy entrées. PMID:23557428

The impact of financial incentives on participants' food purchasing patterns in a supermarket-based randomized controlled trial.

PubMed

Olstad, Dana Lee; Crawford, David A; Abbott, Gavin; McNaughton, Sarah A; Le, Ha Nd; Ni Mhurchu, Cliona; Pollard, Christina; Ball, Kylie

2017-08-25

The impacts of supermarket-based nutrition promotion interventions might be overestimated if participants shift their proportionate food purchasing away from their usual stores. This study quantified whether participants who received price discounts on fruits and vegetables (FV) in the Supermarket Healthy Eating for Life (SHELf) randomized controlled trial (RCT) shifted their FV purchasing into study supermarkets during the intervention period. Participants were 642 females randomly assigned to a 1) skill-building (n = 160), 2) price reduction (n = 161), 3) combined skill-building and price reduction (n = 160), or 4) control (n = 161) group. Participants self-reported the proportion of FV purchased in study supermarkets at baseline, 3- and 6-months post-intervention. Fisher's exact and χ 2 tests assessed differences among groups in the proportion of FV purchased in study supermarkets at each time point. Multinomial logistic regression assessed differences among groups in the change in proportionate FV purchasing over time. Post-intervention, 49% of participants purchased ≥50% of their FV in study supermarkets. Compared to all other groups, the price reduction group was approximately twice as likely (RRR: 1.8-2.2) to have increased proportionate purchasing of FV in study supermarkets from baseline to post-intervention (p< 0.05). Participants who received price reductions on FV were approximately twice as likely to shift their FV purchasing from other stores into study supermarkets during the intervention period. Unless food purchasing data are available for all sources, differential changes in purchasing patterns can make it difficult to discern the true impacts of nutrition interventions. The SHELf trial is registered with Current Controlled Trials Registration ISRCTN39432901, Registered 30 June 2010, Retrospectively registered ( http://www.isrctn.com/ISRCTN39432901 ).

Let them eat fruit! The effect of fruit and vegetable consumption on psychological well-being in young adults: A randomized controlled trial.

PubMed

Conner, Tamlin S; Brookie, Kate L; Carr, Anitra C; Mainvil, Louise A; Vissers, Margreet C M

2017-01-01

This study tested the psychological benefits of a 14-day preregistered clinical intervention to increase fruit and vegetable (FV) consumption in 171 low-FV-consuming young adults (67% female, aged 18-25). Participants were randomly assigned into a diet-as-usual control condition, an ecological momentary intervention (EMI) condition involving text message reminders to increase their FV consumption plus a voucher to purchase FV, or a fruit and vegetable intervention (FVI) condition in which participants were given two additional daily servings of fresh FV to consume on top of their normal diet. Self-report outcome measures were depressive symptoms and anxiety measured pre- and post-intervention, and daily negative and positive mood, vitality, flourishing, and flourishing behaviors (curiosity, creativity, motivation) assessed nightly using a smartphone survey. Vitamin C and carotenoids were measured from blood samples pre- and post-intervention, and psychological expectancies about the benefits of FV were measured post-intervention to test as mediators of psychological change. Only participants in the FVI condition showed improvements to their psychological well-being with increases in vitality, flourishing, and motivation across the 14-days relative to the other groups. No changes were found for depressive symptoms, anxiety, or mood. Intervention benefits were not mediated by vitamin C, carotenoids, or psychological expectancies. We conclude that providing young adults with high-quality FV, rather than reminding them to eat more FV (with a voucher to purchase FV), resulted in significant short-term improvements to their psychological well-being. These results provide initial proof-of-concept that giving young adults fresh fruit and vegetables to eat can have psychological benefits even over a brief period of time. Australian New Zealand Clinical Trials Registry ACTRN12615000183583.

Purification and refolding of anti-T-antigen single chain antibodies (scFvs) expressed in Escherichia coli as inclusion bodies.

PubMed

Yuasa, Noriyuki; Koyama, Tsubasa; Fujita-Yamaguchi, Yoko

2014-02-01

T-antigen (Galβ1-3GalNAcα-1-Ser/Thr) is an oncofetal antigen that is commonly expressed as a carbohydrate determinant in many adenocarcinomas. Since it is associated with tumor progression and metastasis, production of recombinant antibodies specific for T-antigen could lead to the development of cancer diagnostics and therapeutics. Previously, we isolated and characterized 11 anti-T-antigen phage clones from a phage library displaying human single-chain antibodies (scFvs) and purified one scFv protein, 1G11. More recently, we purified and characterized 1E8 scFv protein using a Drosophila S2 expression system. In the current study, four anti-T-antigen scFv genes belonging to Groups 1-4 were purified from inclusion bodies expressed in Escherichia coli cells. Inclusion bodies isolated from E. coli cells were denatured in 3.5 M Gdn-HCl. Solubilized His-tagged scFv proteins were purified using Ni(2+)-Sepharose column chromatography in the presence of 3.5 M Gdn-HCl. Purified scFv proteins were refolded according to a previously published method of step-wise dialysis. Two anti-T-antigen scFv proteins, 1E6 and 1E8 that belong to Groups 1 and 2, respectively, were produced in sufficient amounts, thus allowing further characterization of their binding activity with T-antigen. Specificity and affinity constants determined using enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR), respectively, provided evidence that both 1E8 and 1E6 scFv proteins are T-antigen specific and suggested that 1E8 scFv protein has a higher affinity for T-antigen than 1E6 scFv protein.

A systematic review of methods to assess intake of fruits and vegetables among healthy European adults and children: a DEDIPAC (DEterminants of DIet and Physical Activity) study.

PubMed

Riordan, Fiona; Ryan, Kathleen; Perry, Ivan J; Schulze, Matthias B; Andersen, Lene Frost; Geelen, Anouk; Van't Veer, Pieter; Eussen, Simone; Dagnelie, Pieter; Wijckmans-Duysens, Nicole; Harrington, Janas M

2017-02-01

Evidence suggests that health benefits are associated with consuming recommended amounts of fruits and vegetables (F&V), yet standardised assessment methods to measure F&V intake are lacking. The current review aims to identify methods to assess F&V intake among children and adults in pan-European studies and inform the development of the DEDIPAC (DEterminants of DIet and Physical Activity) toolbox of methods suitable for use in future European studies. A literature search was conducted using three electronic databases and by hand-searching reference lists. English-language studies of any design which assessed F&V intake were included in the review. Studies involving two or more European countries were included in the review. Healthy, free-living children or adults. The review identified fifty-one pan-European studies which assessed F&V intake. The FFQ was the most commonly used (n 42), followed by 24 h recall (n 11) and diet records/diet history (n 7). Differences existed between the identified methods; for example, the number of F&V items on the FFQ and whether potatoes/legumes were classified as vegetables. In total, eight validated instruments were identified which assessed F&V intake among adults, adolescents or children. The current review indicates that an agreed classification of F&V is needed in order to standardise intake data more effectively between European countries. Validated methods used in pan-European populations encompassing a range of European regions were identified. These methods should be considered for use by future studies focused on evaluating intake of F&V.

Let them eat fruit! The effect of fruit and vegetable consumption on psychological well-being in young adults: A randomized controlled trial

PubMed Central

Carr, Anitra C.; Mainvil, Louise A.; Vissers, Margreet C. M.

2017-01-01

This study tested the psychological benefits of a 14-day preregistered clinical intervention to increase fruit and vegetable (FV) consumption in 171 low-FV-consuming young adults (67% female, aged 18–25). Participants were randomly assigned into a diet-as-usual control condition, an ecological momentary intervention (EMI) condition involving text message reminders to increase their FV consumption plus a voucher to purchase FV, or a fruit and vegetable intervention (FVI) condition in which participants were given two additional daily servings of fresh FV to consume on top of their normal diet. Self-report outcome measures were depressive symptoms and anxiety measured pre- and post-intervention, and daily negative and positive mood, vitality, flourishing, and flourishing behaviors (curiosity, creativity, motivation) assessed nightly using a smartphone survey. Vitamin C and carotenoids were measured from blood samples pre- and post-intervention, and psychological expectancies about the benefits of FV were measured post-intervention to test as mediators of psychological change. Only participants in the FVI condition showed improvements to their psychological well-being with increases in vitality, flourishing, and motivation across the 14-days relative to the other groups. No changes were found for depressive symptoms, anxiety, or mood. Intervention benefits were not mediated by vitamin C, carotenoids, or psychological expectancies. We conclude that providing young adults with high-quality FV, rather than reminding them to eat more FV (with a voucher to purchase FV), resulted in significant short-term improvements to their psychological well-being. These results provide initial proof-of-concept that giving young adults fresh fruit and vegetables to eat can have psychological benefits even over a brief period of time. Trial registration: Australian New Zealand Clinical Trials Registry ACTRN12615000183583 PMID:28158239

Capsid-binding retrovirus restriction factors: discovery, restriction specificity and implications for the development of novel therapeutics.

PubMed

Sanz-Ramos, Marta; Stoye, Jonathan P

2013-12-01

The development of drugs against human immunodeficiency virus type 1 infection has been highly successful, and numerous combinational treatments are currently available. However, the risk of the emergence of resistance and the toxic effects associated with prolonged use of antiretroviral therapies have emphasized the need to consider alternative approaches. One possible area of investigation is provided by the properties of restriction factors, cellular proteins that protect organisms against retroviral infection. Many show potent viral inhibition. Here, we describe the discovery, properties and possible therapeutic uses of the group of restriction factors known to interact with the capsid core of incoming retroviruses. This group comprises Fv1, TRIM5α and TRIMCypA: proteins that all act shortly after virus entry into the target cell and block virus replication at different stages prior to integration of viral DNA into the host chromosome. They have different origins and specificities, but share general structural features required for restriction, with an N-terminal multimerization domain and a C-terminal capsid-binding domain. Their overall efficacy makes it reasonable to ask whether they might provide a framework for developing novel antiretroviral strategies.

Fab-dsFv: A bispecific antibody format with extended serum half-life through albumin binding.

PubMed

Davé, Emma; Adams, Ralph; Zaccheo, Oliver; Carrington, Bruce; Compson, Joanne E; Dugdale, Sarah; Airey, Michael; Malcolm, Sarah; Hailu, Hanna; Wild, Gavin; Turner, Alison; Heads, James; Sarkar, Kaushik; Ventom, Andrew; Marshall, Diane; Jairaj, Mark; Kopotsha, Tim; Christodoulou, Louis; Zamacona, Miren; Lawson, Alastair D; Heywood, Sam; Humphreys, David P

2016-10-01

An antibody format, termed Fab-dsFv, has been designed for clinical indications that require monovalent target binding in the absence of direct Fc receptor (FcR) binding while retaining substantial serum presence. The variable fragment (Fv) domain of a humanized albumin-binding antibody was fused to the C-termini of Fab constant domains, such that the VL and VH domains were individually connected to the Cκ and CH1 domains by peptide linkers, respectively. The anti-albumin Fv was selected for properties thought to be desirable to ensure a durable serum half-life mediated via FcRn. The Fv domain was further stabilized by an inter-domain disulfide bond. The bispecific format was shown to be thermodynamically and biophysically stable, and retained good affinity and efficacy to both antigens simultaneously. In in vivo studies, the serum half-life of Fab-dsFv, 2.6 d in mice and 7.9 d in cynomolgus monkeys, was equivalent to Fab'-PEG.

Fab-dsFv: A bispecific antibody format with extended serum half-life through albumin binding

PubMed Central

Davé, Emma; Adams, Ralph; Zaccheo, Oliver; Carrington, Bruce; Compson, Joanne E.; Dugdale, Sarah; Airey, Michael; Malcolm, Sarah; Hailu, Hanna; Wild, Gavin; Turner, Alison; Heads, James; Sarkar, Kaushik; Ventom, Andrew; Marshall, Diane; Jairaj, Mark; Kopotsha, Tim; Christodoulou, Louis; Zamacona, Miren; Lawson, Alastair D.; Heywood, Sam; Humphreys, David P.

2016-01-01

ABSTRACT An antibody format, termed Fab-dsFv, has been designed for clinical indications that require monovalent target binding in the absence of direct Fc receptor (FcR) binding while retaining substantial serum presence. The variable fragment (Fv) domain of a humanized albumin-binding antibody was fused to the C-termini of Fab constant domains, such that the VL and VH domains were individually connected to the Cκ and CH1 domains by peptide linkers, respectively. The anti-albumin Fv was selected for properties thought to be desirable to ensure a durable serum half-life mediated via FcRn. The Fv domain was further stabilized by an inter-domain disulfide bond. The bispecific format was shown to be thermodynamically and biophysically stable, and retained good affinity and efficacy to both antigens simultaneously. In in vivo studies, the serum half-life of Fab-dsFv, 2.6 d in mice and 7.9 d in cynomolgus monkeys, was equivalent to Fab'-PEG. PMID:27532598

The founder of the Friends Foundation--Tessie Soi.

PubMed

Topurua, Ore

2013-01-01

Tessie Soi is well known in Papua New Guinea and beyond for her work with HIV/AIDS (human immunodeficiency virus/acquired immune deficiency syndrome) patients, including through the Friends Foundation, an organization that focuses on helping families affected by HIV and AIDS. This article explores Tessie's early life and childhood, providing insight into some of the values she learned from her parents. Providing details about the Friends Foundation and the Orphan Buddy Systems program, a program Tessie established to support AIDS orphans, the article offers insight into Tessie's beliefs and compassion, simultaneously highlighting the value she places on her family.

A functional role for CD28 costimulation in tumor recognition by single-chain receptor-modified T cells.

PubMed

Moeller, Maria; Haynes, Nicole M; Trapani, Joseph A; Teng, Michele W L; Jackson, Jacob T; Tanner, Jane E; Cerutti, Loretta; Jane, Stephen M; Kershaw, Michael H; Smyth, Mark J; Darcy, Phillip K

2004-05-01

T cells engineered to express single-chain antibody receptors that incorporate TCR-zeta and cluster designation (CD)28 signaling domains (scFv-alpha-erbB2-CD28-zeta) can be redirected in vivo to cancer cells that lack triggering costimulatory molecules. To assess the contribution of CD28 signaling to the function of the scFv-CD28-zeta receptor, we expressed a series of mutated scFv-CD28-zeta receptors directed against erbB2. Residues known to be critical for CD28 signaling were mutated from tyrosine to phenylalanine at position 170 or proline to alanine at positions 187 and 190. Primary mouse T cells expressing either of the mutant receptors demonstrated impaired cytokine (IFN-gamma and GM-CSF) production and decreased proliferation after antigen ligation in vitro and decreased antitumor efficacy in vivo compared with T cells expressing the wild-type scFv-CD28-zeta receptor, suggesting a key signaling role for the CD28 component of the scFv-CD28-zeta receptor. Importantly, cell surface expression, binding capacity and cytolytic activity mediated by the scFv-CD28-zeta receptor were not diminished by either mutation. Overall, this study has definitively demonstrated a functional role for the CD28 component of the scFv-CD28-zeta receptor and has shown that incorporation of costimulatory activity in chimeric scFv receptors is a powerful approach for improving adoptive cancer immunotherapy.

Combined effects of fruit and vegetables intake and physical activity on the risk of metabolic syndrome among Chinese adults.

PubMed

Li, Xin-Tong; Liao, Wei; Yu, Hong-Jie; Liu, Ming-Wei; Yuan, Shuai; Tang, Bo-Wen; Yang, Xu-Hao; Song, Yong; Huang, Yao; Cheng, Shi-le; Chen, Zhi-Yu; Towne, Samuel D; Mao, Zong-Fu; He, Qi-Qiang

2017-01-01

Unbalanced dietary intake and insufficient physical activity (PA) have been recognized as risk factors for metabolic syndrome (MetS). We aimed to examine the independent and combined effects of fruit and vegetables (FV) intake and PA on MetS. A cross-sectional survey was conducted among residents of China in 2009, with fasting blood samples collected. Participants were divided into sufficient/insufficient FV intake and adequate/ inadequate PA groups according to self-reported questionnaires. MetS was defined using the NCEP-ATPIII criteria. The difference of individual MetS components was compared across different PA or FV groups. Multivariable logistic regression was used to assess association between FV/PA and the risk of MetS. A total of 7424 adults were included in the current study. MetS was prevalent in 28.7% of participants, with 24.7% and 32.9% in male and female, respectively. Compared with those with inadequate PA and insufficient FV intake, participants with the combination of adequate PA and sufficient FV intake had the lowest risk of MetS (OR = 0.69,95%CI: 0.59-0.82), following by the group with adequate PA time but insufficient FV intake (OR = 0.74, 95%CI:0.65-0.83). Findings of the current study show that the combination of sufficient FV intake and adequate PA was significantly associated with reduced MetS risk among adult residents of China.

Psychosocial Influences on Fruit and Vegetable Intake Following a NYC Supermarket Discount.

PubMed

Bernales-Korins, Maria; Ang, Ian Yi Han; Khan, Shamima; Geliebter, Allan

2017-08-01

To assess the effect of a 50% discount on fruits and vegetables (F&V) on the purchase and intake of F&V and on psychosocial determinants of F&V intake: self-efficacy (SE), stages of change (SOC), and perceived barriers (PB). This randomized controlled trial was conducted in local supermarkets over 16 weeks, including a 4-week baseline, 8-week discount intervention, and 4-week follow-up. Shoppers with overweight or obesity (BMI > 25) were randomized to receive a discount or no discount via their reward scan card after the baseline. Twenty-four-hour recalls and psychosocial measures were obtained for each study period. Purchases (P < 0.0005) and intakes (P = 0.019) of F&V increased significantly during the intervention, while only F&V intake was sustained at follow-up. The discount intervention increased SE (P < 0.01) and SOC (P < 0.05) and did not decrease PB (P = 0.057) during the intervention. SOC mediated the discount intervention effect on F&V intake (P < 0.05) during the intervention, explaining 43% of variance. A supermarket discount intervention led to increases in purchases and intakes of F&V and increases in the psychosocial factors SE and SOC and did not decrease PB. The discount intervention prompted participants to move from the preparation to action stage of SOC, which acted as a mediator for increased F&V intake. © 2017 The Obesity Society.

Downregulation of Extracellular Matrix Metalloproteinase Inducer by scFv-M6-1B9 Intrabody Suppresses Cervical Cancer Invasion Through Inhibition of Urokinase-Type Plasminogen Activator.

PubMed

Panich, Tipattaraporn; Tragoolpua, Khajornsak; Pata, Supansa; Tayapiwatana, Chatchai; Intasai, Nutjeera

2017-02-01

Overexpression of extracellular matrix metalloproteinase inducer (EMMPRIN) accelerates tumor invasion and metastasis via activation of matrix metalloproteinases (MMPs) and urokinase-type plasminogen activator (uPA) expression. The authors were interested in whether the scFv-M6-1B9 intrabody against EMMPRIN that retains EMMPRIN in endoplasmic reticulum could be a potential tool to suppress cervical cancer invasion through inhibition of uPA. The chimeric adenoviral vector Ad5/F35-scFv-M6-1B9 was transferred into human cervical carcinoma HeLa cells to produce the scFv-M6-1B9 intrabody against EMMPRIN. Cell surface expression of EMMPRIN, the membrane-bound uPA, the enzymatic activity of secreted uPA, and the invasion ability were analyzed. The scFv-M6-1B9 intrabody successfully diminished the cell surface expression of EMMPRIN and the membrane-bound uPA on HeLa cells. uPA activity from tissue culture media of EMMPRIN-downregulated HeLa cells was decreased. The invasion ability of HeLa cells harboring scFv-M6-1B9 intrabody was also suppressed. These results suggested that the scFv-M6-1B9 intrabody might represent a potential approach for invasive cervical cancer treatment. The application of scFv-M6-1B9 intrabody in animal experiments and preclinical studies would be investigated further.

Magnetic Resonance Venous Volume Measurements in Peripheral Artery Disease (from ELIMIT).

PubMed

Kamran, Hassan; Nambi, Vijay; Negi, Smita; Yang, Eric Y; Chen, Changyi; Virani, Salim S; Kougias, Panos; Lumsden, Alan B; Morrisett, Joel D; Ballantyne, Christie M; Brunner, Gerd

2016-11-01

The relation between the arterial and venous systems in patients with impaired lower extremity blood flow remains poorly described. The objective of this secondary analysis of the Effectiveness of Intensive Lipid Modification Medication in Preventing the Progression on Peripheral Artery Disease Trial was to determine the association between femoral vein (FV) volumes and measurements of peripheral artery disease. FV wall, lumen, and total volumes were quantified with fast spin-echo proton density-weighted magnetic resonance imaging scans in 79 patients with peripheral artery disease over 2 years. Reproducibility was excellent for FV total vessel (intraclass correlation coefficient 0.924, confidence interval 0.910 to 0.935) and lumen volumes (intraclass correlation coefficient 0.893, confidence interval 0.873 to 0.910). Baseline superficial femoral artery volumes were directly associated with FV wall (r = 0.46, p <0.0001), lumen (r = 0.42, p = 0.0001), and total volumes (r = 0.46, p <0.0001). The 2-year change in maximum walking time was inversely associated with the 24-month change in FV total volume (r = -0.45, p = 0.03). In conclusion, FV volumes can be measured reliably with fast spin-echo proton density-weighted magnetic resonance imaging, and baseline superficial femoral artery plaque burden is positively associated with FV volumes, whereas the 2-year change in FV volumes and leg function show an inverse relation. Copyright © 2016 Elsevier Inc. All rights reserved.

The impact of fruit and vegetable intake on weight management

USDA-ARS?s Scientific Manuscript database

Fruit and vegetables (FV) are important sources of phytochemicals, dietary fiber, and low energy density, and their consumption may be protective against obesity. Despite these potential benefits of FV consumption on human health, rates of FV intake remain low throughout the world. This chapter revi...

Relationship of fruit and vegetable intake with adiposity: a systematic review

USDA-ARS?s Scientific Manuscript database

Fruit and vegetable (FV) intake has been proposed to protect against obesity. The purpose of this paper was to assess the FV consumption to adiposity relationship. Twenty-three publications were included. Inclusion criteria: longitudinal or experimental designs; FV intake tested in relation to adipo...

Phage display antibodies against ectromelia virus that neutralize variola virus: Selection and implementation for p35 neutralizing epitope mapping.

PubMed

Khlusevich, Yana; Matveev, Andrey; Baykov, Ivan; Bulychev, Leonid; Bormotov, Nikolai; Ilyichev, Ivan; Shevelev, Georgiy; Morozova, Vera; Pyshnyi, Dmitrii; Tikunova, Nina

2018-04-01

In this study, five phage display antibodies (pdAbs) against ectromelia virus (ECTV) were selected from vaccinia virus (VACV)-immune phage-display library of human single chain variable fragments (scFv). ELISA demonstrated that selected pdAbs could recognize ECTV, VACV, and cowpox virus (CPXV). Atomic force microscopy visualized binding of the pdAbs to VACV. Three of the selected pdAbs neutralized variola virus (VARV) in the plaque reduction neutralization test. Western blot analysis of ECTV, VARV, VACV, and CPXV proteins indicated that neutralizing pdAbs bound orthopoxvirus 35 kDa proteins, which are encoded by the open reading frames orthologous to the ORF H3L in VACV. The fully human antibody fh1A was constructed on the base of the VH and VL domains of pdAb, which demonstrated a dose-dependent inhibition of plaque formation after infection with VARV, VACV, and CPXV. To determine the p35 region responsible for binding to neutralizing pdAbs, a panel of truncated p35 proteins was designed and expressed in Escherichia coli cells, and a minimal p35 fragment recognized by selected neutralizing pdAbs was identified. In addition, peptide phage-display combinatorial libraries were applied to localize the epitope. The obtained data indicated that the epitope responsible for recognition by the neutralizing pdAbs is discontinuous and amino acid residues located within two p35 regions, 15-19 aa and 232-237 aa, are involved in binding with neutralizing anti-p35 antibodies. Copyright © 2018. Published by Elsevier B.V.

Influences on children's dietary behavior, and innovative attempts to change it

USDA-ARS?s Scientific Manuscript database

Fruit and vegetable (FV) intake may protect against several chronic diseases, and the preferences and habits in relation to FV intake appear to form in early childhood. Child FV intake reflects many influences from multiple levels (e.g., internal to the child, family, school, neighborhood). We have ...

77 FR 67673 - Fernando Valle, M.D.; Decision and Order

Federal Register 2010, 2011, 2012, 2013, 2014

2012-11-13

... DEPARTMENT OF JUSTICE Drug Enforcement Administration [Docket No. 12-56] Fernando Valle, M.D... Certificate of Registration Numbers FV1935595, FV2000711, and FV2000735, issued to Fernando Valle, M.D., be, and they hereby are, revoked. I further order that any pending applications of Fernando Valle, M.D...

Molecular Exploration of Beta-Lactamases in Fusarium verticillioides

USDA-ARS?s Scientific Manuscript database

The mycotoxigenic fungus Fusarium verticillioides (Fv) is one of the most prevalent maize fungal pathogens. Fv mycotoxins are a significant food safety issue and have given rise to exposure concerns worldwide. The FDB1 locus, a beta-lactamase-containing Fv gene cluster, was previously shown to be in...

Gallium-68-labeled anti-HER2 single-chain Fv fragment: development and in vivo monitoring of HER2 expression.

PubMed

Ueda, Masashi; Hisada, Hayato; Temma, Takashi; Shimizu, Yoichi; Kimura, Hiroyuki; Ono, Masahiro; Nakamoto, Yuji; Togashi, Kaori; Saji, Hideo

2015-02-01

We aimed to develop a gallium-68 (Ga-68)-labeled single-chain variable fragment (scFv) targeting the human epidermal growth factor receptor 2 (HER2) to rapidly and noninvasively evaluate the status of HER2 expression. Anti-HER2 scFv was labeled with Ga-68 by using deferoxamine (Df) as a bifunctional chelate. Biodistribution of [(68)Ga]Df-anti-HER2 scFv was examined with tumor-bearing mice and positron emission tomography (PET) imaging was performed. The changes in HER2 expression after anti-HER2 therapy were monitored by PET imaging. [(68)Ga]Df-anti-HER2 scFv was obtained with high radiochemical yield after only a 5-min reaction at room temperature. The probe showed high accumulation in HER2-positive xenografts and the intratumoral distribution of radioactivity coincided with HER2-positive regions. Furthermore, [(68)Ga]Df-anti-HER2 scFv helped visualize HER2-positive xenografts and monitor the changes in HER2 expression after anti-HER2 therapy. [(68)Ga]Df-anti-HER2 scFv could be a promising probe to evaluate HER2 status by in vivo PET imaging, unless trastuzumab is prescribed as part of the therapy.

Evolution and manipulation of parasitoid egg load.

PubMed

Gandon, Sylvain; Varaldi, Julien; Fleury, Frédéric; Rivero, Ana

2009-11-01

In proovigenic parasitoids such as Leptopilina boulardi, the female emerges with a limited egg load and no further eggs are produced during its adult life. A female thus runs the risk of exhausting this limited supply of eggs before the end of her life. Given that the production of an egg is costly, what is the evolutionarily stable egg load at emergence? This question has attracted a lot of attention in the last decade. Here, we analyze a model that allows us to track both the evolution and the population dynamics of a solitary, proovigenic parasitoid. First, we show how host-parasitoid dynamics feedbacks on the evolution of parasitoid egg load. Second, we use this model to consider the situation in which the parasitoid can be infected by a virus that manipulates the oviposition behavior of the females. In particular, we model the effect of the LbFV virus in L. boulardi, a virus that is known to enhance its horizontal transmission by increasing superparasitism (i.e., the laying of eggs in a host already parasitized). Specifically, we model (1) the effect of the virus on parasitoid egg load strategies, and (2) the evolution of egg load manipulation by the virus. This analysis yields two alternative, yet not mutually exclusive, adaptive explanations for the observation that females infected by the virus harbor higher egg loads than uninfected females. Infected females could either respond plastically to the infection status, or be manipulated by the virus. Further experimental work is required to distinguish between these two hypotheses. In a broader context, we present a general theoretical framework that allows us to study the epidemiology, the evolution, the coevolution, and the evolution of manipulation of various reproductive strategies of parasitoids.

Evaluation of (1R,2R)-1-(5'-methylfur-3'-yl)propane-1,2,3-triol, a sphydrofuran derivative isolated from a Streptomyces species, as an anti-herpesvirus drug.

PubMed

Hayashi, K; Kawahara, K; Nakai, C; Sankawa, U; Seto, H; Hayashi, T

2000-08-01

(1R,2R)-1-(5'-Methylfur-3'-yl)propane-1,2,3-triol (MFPT), a stable anhydro derivative of sphydrofuran, was obtained from the culture broth of STREPTOMYCES: sp. strain FV60 as an inhibitor of herpes simplex virus type 1 (HSV-1). The compound showed antiherpetic activity with a 50% inhibitory concentration of 1.2 IM in an in vitro assay system. Although the binding of virus to host cells was not inhibited, the penetration of virus into cells was moderately blocked by MFPT. Some of the viruses, once they had penetrated cells, failed to form plaques in the presence of MFPT. When added to the late stages of HSV-1 replication, MFPT also inhibited virus production. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis analysis of isotope-labelled HSV-specific proteins revealed that a protein or proteins with reduced molecular weight (about 120 kDa) was clearly detected in cells treated with MFPT. Western blot analysis with antibodies against three HSV-specific glycoproteins (gB, gC and gD) showed a significant difference in gC synthesis between untreated and MFPT-treated cells. Release of progeny viruses was suppressed by MFPT. Syncytium formation by HSV-1 strain HF was inhibited and small plaques with rounded cells were formed in MFPT-treated cell cultures. When wild-type HSV-1 was serially propagated under the selective pressure of MFPT, resistant virus emerged. MFPT-resistant progeny were accompanied by the formation of plaques with rounded cells. These results, taken together, suggest that MFPT might act by limiting the maturation of HSV-specific glycoproteins, particularly of HSV-1 gC.

One-Shot In Vitro Evolution Generated an Antibody Fragment for Testing Urinary Cotinine with More Than 40-Fold Enhanced Affinity.

PubMed

Oyama, Hiroyuki; Morita, Izumi; Kiguchi, Yuki; Banzono, Erika; Ishii, Kasumi; Kubo, Satoshi; Watanabe, Yoshiro; Hirai, Anna; Kaede, Chiaki; Ohta, Mitsuhiro; Kobayashi, Norihiro

2017-01-03

Immunoassays for cotinine, a major nicotine metabolite, in the urine are useful for monitoring the degree of tobacco smoke exposure. However, hybridoma-based anti-cotinine antibodies lack sufficient binding affinity to perform practically sensitive measurements, and thus most cotinine assays still rely on polyclonal antibodies. Here, we describe the generation of a mutant single-chain Fv fragment (scFv) that was used in an enzyme-linked immunosorbent assay (ELISA) to determine urinary cotinine levels in passive smokers. A "wild-type" scFv (scFv-wt) with a K a value of 2.7 × 10 7 M -1 (at 4 °C) was prepared by linking the V H and V L domains in a mouse anti-cotinine antibody. "One-shot" random mutagenesis on the scFv-wt gene by error-prone PCR generated mutant scFv genes, which were expressed on phage particles. Repeated panning directed toward mutants with slower off-rates selected scFv clones that showed improved sensitivity in an ELISA system. One of these mutants (scFv#m1-54) with five amino acid substitutions showed more than a 40-fold enhanced K a (1.2 × 10 9 M -1 at 4 °C) and, thus, was used to monitor human urinary cotinine. A limited amount of soluble scFv was reacted with urine specimens (or cotinine standards) at 4 °C for 120 min in microwells on which cotinine residues had been immobilized. The midpoint of the dose-response curves under optimized conditions (0.27 ng/assay) was more than 100-fold lower than the ELISA results obtained using scFv-wt. The limit of detection (8.4 pg/assay) corresponded to 0.17 ng/mL urinary cotinine, which was satisfactorily low for testing the threshold levels for passive smoke exposure. The assay values for volunteers correlated with the values determined using a commercial assay kit. This study evidently showed the potential of a molecular breeding approach, in which simple in vitro evolution might generate superior antibody reagents as cloned proteins, overcoming the limited molecular diversity inherent to conventional immunization-based antibodies.

Using Virtual Pets to Increase Fruit and Vegetable Consumption in Children: A Technology-Assisted Social Cognitive Theory Approach.

PubMed

Ahn, Sun Joo Grace; Johnsen, Kyle; Moore, James; Brown, Scott; Biersmith, Melanie; Ball, Catherine

2016-02-01

A virtual pet in the form of a mid-sized dog was developed based on the framework of social cognitive theory and tested as a vehicle for promoting fruit and vegetable (F&V) consumption in children. Three groups of children (N = 68) between the ages of 7 and 13 years were studied: baseline (no treatment), computer only, and virtual dog. Children in the virtual dog condition interacted with the virtual dog for 3 days, setting F&V consumption goals and receiving evaluation and reinforcement based on whether they met their self-set goals. Children vicariously experienced future health outcomes of F&V consumption by seeing, hearing, and feeling their virtual dog's physical and mental health improve or deteriorate based on their F&V consumption in the physical world. Children in the computer only condition interacted with a computer system that presented equivalent features, but without the virtual dog. Children in the baseline condition did not receive any experimental treatment. Results indicated that children in the virtual dog condition chose to be served significantly more F&V than those in the computer only or baseline conditions did. However, children in the virtual dog condition were unable to consume significantly more F&V than those in the computer only condition, although children in those two conditions consumed more F&V than the baseline condition. Food preferences did not differ significantly across the three conditions before and after the experimental treatments. Theoretical and practical potentials of using a virtual pet to promote F&V consumption systematically in children are discussed.

Generation of a Highly Reactive Chicken-Derived Single-Chain Variable Fragment against Fusarium verticillioides by Phage Display

PubMed Central

Hu, Zu-Quan; Liu, Jin-Long; Li, He-Ping; Xing, Shu; Xue, Sheng; Zhang, Jing-Bo; Wang, Jian-Hua; Nölke, Greta; Liao, Yu-Cai

2012-01-01

Fusarium verticillioides is the primary causal agent of Fusarium ear and kernel rot in maize, producing fumonisin mycotoxins that are toxic to humans and domestic animals. Rapid detection and monitoring of fumonisin-producing fungi are pivotally important for the prevention of mycotoxins from entering into food/feed products. Chicken-derived single-chain variable fragments (scFvs) against cell wall-bound proteins from F. verticillioides were isolated from an immunocompetent phage display library. Comparative phage enzyme-linked immunosorbant assays (ELISAs) and sequencing analyses identified four different scFv antibodies with high sensitivity. Soluble antibody ELISAs identified two highly sensitive scFv antibodies, FvCA3 and FvCA4, with the latter being slightly more sensitive. Three-dimensional modeling revealed that the FvCA4 may hold a better overall structure with CDRH3, CDRL1 and CDRL3 centered in the core region of antibody surface compared with that of other scFvs. Immunofluorescence labeling revealed that the binding of FvCA4 antibody was localized to the cell walls of conidiospores and hyphae of F. verticillioides, confirming the specificity of this antibody for a surface target. This scFv antibody was able to detect the fungal mycelium as low as 10−2 μg/mL and contaminating mycelium at a quantity of 10−2 mg/g maize. This is the first report that scFv antibodies derived from phage display have a wide application for rapid and accurate detection and monitoring of fumonisin-producing pathogens in agricultural samples. PMID:22837678

Dietary Foeniculum vulgare Mill extract attenuated UVB irradiation-induced skin photoaging by activating of Nrf2 and inhibiting MAPK pathways.

PubMed

Sun, Zhengwang; Park, Sang Yong; Hwang, Eunson; Park, Bom; Seo, Seul A; Cho, Jin-Gyeong; Zhang, Mengyang; Yi, Tae-Hoo

2016-11-15

Foeniculum vulgare Mill (FV) has long been prescribed in traditional medicine due to its antioxidant anti-inflammatory properties. However, little research has been done on the use of FV to alleviate changes in UVB-induced photoaging PURPOSE: This study was to investigate the photoprotective effects and mechanism of FV in vitro and in vivo. The anti-photoaging effect of FV was assessed in normal human dermal fibroblasts (NHDFs) in vitro. The secretion of reactive oxygen species (ROS), lactate dehydrogenase (LDH), GSH, matrix metalloproteinases (MMPs), procollagen type I, IL-6 and transforming growth factor-β1 (TGF-β1) were measured by kits. Additionally, the level of nuclear factor (erythroid-derived 2)-like 2 (Nrf2), p-ERK and p38 were evaluated by western blotting. In vivo, H&E and Masson's trichrome staining were employed. The expression of MMP-1, procollagen type I, TGF-β1 and elastin were measured by western blot. FV significantly increased the production of collagen, elastin and TGF-β1 levels, while blocked matrix metalloproteinases (MMPs) production in UVB irradiation induced hairless mice, which were consistent with the result in NHDFs. Furthermore, FV dose-dependently decreased the production of ROS and LDH by promoting the nuclear amount of Nrf2 and enhancing the expression of cytoprotective antioxidants such as GSH. FV also significantly quenched UVB-induced phosphorylation of ERK and p38 in NHDFs. Our results indicate that FV is a potential botanical agent for the treatment of skin damage induced by UV irradiation. Copyright © 2016 Elsevier GmbH. All rights reserved.

Production and characterization of a single chain variable fragment (scFv) for the mycotoxin deoxynivalenol

USDA-ARS?s Scientific Manuscript database

Deoxynivalenol (DON)is a mycotoxin produced by certain fungi that infest cereal grains worldwide. A hybridoma cell line producing a monoclonal antibody (Mab) recognizing DON was used as the starting point in the development of a recombinant single chain variable fragment (scFv) antibody. The scFv wa...

Children's Daily Fruit and Vegetable Intake: Associations with Maternal Intake and Child Weight Status

ERIC Educational Resources Information Center

Miller, Paige; Moore, Renee H.; Kral, Tanja V. E.

2011-01-01

Objective: To evaluate associations between children's and their mothers' fruit and vegetable (FV) intake and children's FV intake and weight status. Methods: Mothers (n = 39) residing in Philadelphia, PA completed a subsection of the Diet History Questionnaire assessing their FV intake. Mothers also completed this questionnaire to estimate FV…

75 FR 37288 - Kiwifruit Grown in California; Order Amending Marketing Order No. 920

Federal Register 2010, 2011, 2012, 2013, 2014

2010-06-29

... DEPARTMENT OF AGRICULTURE Agricultural Marketing Service 7 CFR Part 920 [Doc. No. AO-FV-08-0174; AMS-FV-08-0085; FV08-920-3] Kiwifruit Grown in California; Order Amending Marketing Order No. 920 AGENCY: Agricultural Marketing Service, USDA. ACTION: Final rule. SUMMARY: This rule amends Marketing...

76 FR 4201 - Kiwifruit Grown in California; Order Amending Marketing Order No. 920; Correction

Federal Register 2010, 2011, 2012, 2013, 2014

2011-01-25

... DEPARTMENT OF AGRICULTURE Agricultural Marketing Service 7 CFR Part 920 [Doc. No. AO-FV-08-0174; AMS-FV-08-0085; FV08-920-3 C] Kiwifruit Grown in California; Order Amending Marketing Order No. 920; Correction AGENCY: Agricultural Marketing Service, USDA. ACTION: Correcting amendment. SUMMARY: This document...

Validating the Hamilton Anatomy of Risk Management-Forensic Version and the Aggressive Incidents Scale.

PubMed

Cook, Alana N; Moulden, Heather M; Mamak, Mini; Lalani, Shams; Messina, Katrina; Chaimowitz, Gary

2018-06-01

The Hamilton Anatomy of Risk Management-Forensic Version (HARM-FV) is a structured professional judgement tool of violence risk developed for use in forensic inpatient psychiatric settings. The HARM-FV is used with the Aggressive Incidents Scale (AIS), which provides a standardized method of recording aggressive incidents. We report the findings of the concurrent validity of the HARM-FV and the AIS with widely used measures of violence risk and aggressive acts, the Historical, Clinical, Risk Management-20, Version 3 (HCR-20 V3 ) and a modified version of the Overt Aggression Scale. We also present findings on the predictive validity of the HARM-FV in the short term (1-month follow-up periods) for varying severities of aggressive acts. The results indicated strong support for the concurrent validity of the HARM-FV and AIS and promising support for the predictive accuracy of the tool for inpatient aggression. This article provides support for the continued clinical use of the HARM-FV within an inpatient forensic setting and highlights areas for further research.

Fast conversion of scFv to Fab antibodies using type IIs restriction enzymes.

PubMed

Sanmark, Hanna; Huovinen, Tuomas; Matikka, Tero; Pettersson, Tiina; Lahti, Maria; Lamminmäki, Urpo

2015-11-01

Single chain variable fragment (scFv) antibody libraries are widely used for developing novel bioaffinity reagents, although Fab or IgG molecules are the preferred antibody formats in many final applications. Therefore, rapid conversion methods for combining multiple DNA fragments are needed to attach constant domains to the scFv derived variable domains. In this study we describe a fast and easy cloning method for the conversion of single framework scFv fragments to Fab fragments using type IIS restriction enzymes. All cloning steps excluding plating of the Fab transformants can be done in 96 well plates and the procedure can be completed in one working day. The concept was tested by converting 69 scFv clones into Fab format on 96 well plates, which resulted in 93% success rate. The method is particularly useful as a high-throughput tool for the conversion of the chosen scFv clones into Fab molecules in order to analyze them as early as possible, as the conversion can significantly affect the binding properties of the chosen clones. Copyright © 2015 Elsevier B.V. All rights reserved.

[A report of two Chinese familial Budd-Chiari syndrome].

PubMed

Feng, B; Xu, K; Jiang, H; Fu, W; Li, H; Guo, M; Liu, X; Wang, Z

2000-09-01

To investigate the etiology of two Chinese familial Budd-Chiari syndrome (BCS). Four patients with familial BCS (from A and B families), and the other 41 family members were detected by angiography, ultrasound Dopler, etiology analysis and Factor V Leiden (FvL) mutation analysis. Four BCS patients were proved by angiography, 2 by ultra sound Dopler in family A. Ten members in family A were varicosis in low extremeties. FvL mutation was detected in 4 of 6 patients and 2 normal family members. A II(2), A III(7, 11, 15,) B II(10) and B III(5) had FvL mutation. The FvL mutations were compatible with Mendel hereditary law. FvL mutation may be one of main risk factors and varicosis in low extremeties may be another risk factors for familial BCS.

Types of fruits and vegetables used in commercial baby foods and their contribution to sugar content.

PubMed

Garcia, Ada Lizbeth; McLean, Kimberley; Wright, Charlotte M

2016-10-01

Fruits and vegetables (F&V) are often featured in names of commercial baby foods (CBFs). We aimed to survey all available CBFs in the UK market with F&V included in the food name in order to describe the amount and types of F&V used in CBF and their contribution to total sugar content. Food labels were used to identify F&V and total sugar content. Fruits were more common than vegetables in names of the 329 CBFs identified. The six most common F&V in the names were all relatively sweet: apple, banana, tomato, mango, carrot and sweet potato. The percentage of F&V in the foods ranged from a median of 94% for sweet-spoonable to 13% for dry-savoury products. Fruit content of sweet foods (n = 177) was higher than vegetable content of savoury foods (n = 152) with a median (IQR) of 64.0 g/100 g (33.0-100.0) vs. 46.0 g/100 g (33-56.7). Fruit juice was added to 18% of products. The proportion of F&V in CBF correlated significantly with sugar content for all the food types except dry-savoury food (sweet-spoonable r = 0.24, P = 0.006; savoury-spoonable r = 0.65, P < 0.001; sweet-dry r = 0.81, P < 0.001; savoury-dry r = 0.51, P = 0.06) and explained up to two-thirds of the variation in sugar content. The F&V content of CBFs mainly consists of fruits and relatively sweet vegetables which are unlikely to encourage preferences for bitter-tasting vegetables or other non-sweet foods. F&V contribute significantly to the total sugar content, particularly of savoury foods. © 2015 John Wiley & Sons Ltd.

Platelets Contain Tissue Factor Pathway Inhibitor-2 Derived from Megakaryocytes and Inhibits Fibrinolysis*

PubMed Central

Vadivel, Kanagasabai; Ponnuraj, Sathya-Moorthy; Kumar, Yogesh; Zaiss, Anne K.; Bunce, Matthew W.; Camire, Rodney M.; Wu, Ling; Evseenko, Denis; Herschman, Harvey R.; Bajaj, Madhu S.; Bajaj, S. Paul

2014-01-01

Tissue factor pathway inhibitor-2 (TFPI-2) is a homologue of TFPI-1 and contains three Kunitz-type domains and a basic C terminus region. The N-terminal domain of TFPI-2 is the only inhibitory domain, and it inhibits plasma kallikrein, factor XIa, and plasmin. However, plasma TFPI-2 levels are negligible (≤20 pm) in the context of influencing clotting or fibrinolysis. Here, we report that platelets contain significant amounts of TFPI-2 derived from megakaryocytes. We employed RT-PCR, Western blotting, immunohistochemistry, and confocal microscopy to determine that platelets, MEG-01 megakaryoblastic cells, and bone marrow megakaryocytes contain TFPI-2. ELISA data reveal that TFPI-2 binds factor V (FV) and partially B-domain-deleted FV (FV-1033) with Kd ∼9 nm and binds FVa with Kd ∼100 nm. Steady state analysis of surface plasmon resonance data reveal that TFPI-2 and TFPI-1 bind FV-1033 with Kd ∼36–48 nm and bind FVa with Kd ∼252–456 nm. Further, TFPI-1 (but not TFPI-1161) competes with TFPI-2 in binding to FV. These data indicate that the C-terminal basic region of TFPI-2 is similar to that of TFPI-1 and plays a role in binding to the FV B-domain acidic region. Using pull-down assays and Western blots, we show that TFPI-2 is associated with platelet FV/FVa. TFPI-2 (∼7 nm) in plasma of women at the onset of labor is also, in part, associated with FV. Importantly, TFPI-2 in platelets and in plasma of pregnant women inhibits FXIa and tissue-type plasminogen activator-induced clot fibrinolysis. In conclusion, TFPI-2 in platelets from normal or pregnant subjects and in plasma from pregnant women binds FV/Va and regulates intrinsic coagulation and fibrinolysis. PMID:25262870

High red meat intake and all-cause cardiovascular and cancer mortality: is the risk modified by fruit and vegetable intake?

PubMed

Bellavia, Andrea; Stilling, Frej; Wolk, Alicja

2016-10-01

High red meat consumption is associated with a shorter survival and higher risk of cardiovascular disease (CVD), cancer, and all-cause mortality. Fruit and vegetable (FV) consumption is associated with a longer survival and lower mortality risk. Whether high FV consumption can counterbalance the negative impact of high red meat consumption is unknown. We evaluated 2 large prospective cohorts of Swedish men and women (the Swedish Mammography Cohort and the Cohort of Swedish Men) to determine whether the association between red meat consumption and the risk of all-cause, CVD, and cancer-specific mortality differs across amounts of FV intake. The study population included 74,645 Swedish men and women. Red meat and FV consumption were assessed through a self-administered questionnaire. We estimated HRs of all-cause, CVD, and cancer mortality according to quintiles of total red meat consumption. We next investigated possible interactions between red meat and FV consumption and evaluated the dose-response associations at low, medium, and high FV intake. Compared with participants in the lowest quintile of total red meat consumption, those in the highest quintile had a 21% increased risk of all-cause mortality (HR: 1.21; 95% CI: 1.13, 1.29), a 29% increased risk of CVD mortality (HR: 1.29; 95% CI: 1.14, 1.46), and no increase in the risk of cancer mortality (HR: 1.00; 95% CI: 0.88, 1.43). Results were remarkably similar across amounts of FV consumption, and no interaction between red meat and FV consumption was detected. High intakes of red meat were associated with a higher risk of all-cause and CVD mortality. The increased risks were consistently observed in participants with low, medium, and high FV consumption. The Swedish Mammography Cohort and the Cohort of Swedish Men were registered at clinicaltrials.gov as NCT01127698 and NCT01127711, respectively. © 2016 American Society for Nutrition.

Mid-childhood fruit and vegetable consumption: The roles of early liking, early consumption, and maternal consumption.

PubMed

Kong, Kai Ling; Gillman, Matthew W; Rifas-Shiman, Sheryl L; Wen, Xiaozhong

2016-10-01

Previous studies have shown that early liking, early consumption, and maternal consumption of fruits and vegetables (F&V) each predict children's F&V consumption, but no one has examined the independent contributions of these three correlated factors. We aim to examine the extent to which each of these 3 factors is associated with F&V consumption in mid-childhood after accounting for the other 2 in the analysis. We analyzed data from 901 mother-child dyads from Project Viva, a prospective pre-birth cohort study. Mothers reported their child's early liking and consumption of F&V at age 2 years and later consumption at mid-childhood (median age 7.7 years). They also reported their own consumption of F&V at 6 months postpartum. We used multivariable linear regression models to examine the independent associations of these 3 factors with mid-childhood consumption, adjusting for socio-demographic, pregnancy, and child confounders. At 2 years, 53% of the mothers strongly agreed that their child liked fruit and 25% strongly agreed that their child liked vegetables. F&V consumption was 2.5 (1.3) and 1.8 (1.1) times/d at age 2 y and 1.5 (1.1) and 1.3 (0.8) times/d in mid-childhood. Maternal F&V consumption was 1.4 (1.1) and 1.5 (1.0) times/d, respectively. Children's early consumption played the most predominant role. For every 1 time/d increment in children's early consumption of F&V, mid-childhood consumption was higher by 0.25 (95% confidence interval [CI]: 0.19, 0.30) times/d for fruits and 0.21 (95% CI: 0.16, 0.26) times/d for vegetables, adjusted for confounders plus the other 2 exposures. In conclusion, children's early F&V consumption has the most significant influence on children's later consumption. Published by Elsevier Ltd.

Gardening Experience Is Associated with Increased Fruit and Vegetable Intake among First-Year College Students: A Cross-Sectional Examination.

PubMed

Loso, Jennifer; Staub, Daniel; Colby, Sarah E; Olfert, Melissa D; Kattelmann, Kendra; Vilaro, Melissa; Colee, James; Zhou, Wenjun; Franzen-Castle, Lisa; Mathews, Anne E

2018-02-01

Gardening interventions have been shown to increase fruit and vegetable (F/V) intake among school-aged children. It is unknown whether these effects persist into later adolescence or adulthood, and little is known about whether gardening in later adolescence is related to F/V intake. To identify the relationship between both childhood and recent (within the past 12 months) gardening experiences and current F/V intake among college students. A cross-sectional evaluation of 1,121 college freshmen with suboptimal F/V consumption from eight US universities. Participants completed the National Cancer Institute Fruit and Vegetable Screener and questions about gardening experiences. Respondents were grouped as having gardened or not gardened during childhood and recently. A linear mixed model was used to evaluate the relationship between childhood and recent gardening and current F/V intake. Of the student participants, 11% reported gardening only during childhood, 19% reported gardening only recently, 20% reported gardening both as a child and recently, and 49% of students reported never having gardened. Students who gardened both during childhood and recently had a significantly higher mean current intake of F/V compared with students who never gardened (2.5±0.6 vs 1.9±0.5 cup equivalents [CE], respectively; P<0.001). In addition, F/V intake increased with frequency of recent gardening engagement when comparing students who did not garden with those who gardened monthly or weekly (2.1±0.5 CE, 2.4±0.6 CE, and 2.8±0.7 CE, respectively; P<0.001). This analysis suggests that the combination of childhood and recent gardening experience is associated with greater current F/V intake among first-year college students not currently meeting national F/V recommendations. In addition, a greater frequency of gardening experience may further enhance this effect. Copyright © 2018 Academy of Nutrition and Dietetics. Published by Elsevier Inc. All rights reserved.

A quantile regression approach can reveal the effect of fruit and vegetable consumption on plasma homocysteine levels.

PubMed

Verly, Eliseu; Steluti, Josiane; Fisberg, Regina Mara; Marchioni, Dirce Maria Lobo

2014-01-01

A reduction in homocysteine concentration due to the use of supplemental folic acid is well recognized, although evidence of the same effect for natural folate sources, such as fruits and vegetables (FV), is lacking. The traditional statistical analysis approaches do not provide further information. As an alternative, quantile regression allows for the exploration of the effects of covariates through percentiles of the conditional distribution of the dependent variable. To investigate how the associations of FV intake with plasma total homocysteine (tHcy) differ through percentiles in the distribution using quantile regression. A cross-sectional population-based survey was conducted among 499 residents of Sao Paulo City, Brazil. The participants provided food intake and fasting blood samples. Fruit and vegetable intake was predicted by adjusting for day-to-day variation using a proper measurement error model. We performed a quantile regression to verify the association between tHcy and the predicted FV intake. The predicted values of tHcy for each percentile model were calculated considering an increase of 200 g in the FV intake for each percentile. The results showed that tHcy was inversely associated with FV intake when assessed by linear regression whereas, the association was different when using quantile regression. The relationship with FV consumption was inverse and significant for almost all percentiles of tHcy. The coefficients increased as the percentile of tHcy increased. A simulated increase of 200 g in the FV intake could decrease the tHcy levels in the overall percentiles, but the higher percentiles of tHcy benefited more. This study confirms that the effect of FV intake on lowering the tHcy levels is dependent on the level of tHcy using an innovative statistical approach. From a public health point of view, encouraging people to increase FV intake would benefit people with high levels of tHcy.

Location of School Lunch Salad Bars and Fruit and Vegetable Consumption in Middle Schools: A Cross-Sectional Plate Waste Study.

PubMed

Adams, Marc A; Bruening, Meg; Ohri-Vachaspati, Punam; Hurley, Jane C

2016-03-01

The school lunch environment is a prime target for increasing a child's consumption of fresh fruits and vegetables (F/V). Salad bars are heavily encouraged in schools; however, more research is needed to examine the contexts in which salad bars promote consumption of F/V among students. To compare the amount of fresh F/V self-served, consumed, and wasted by students during lunch at schools with differing salad bar placement: inside or outside of the serving line. Cross-sectional plate waste study in which salad bar placement differed between schools. A random sample of middle school students (N=533) from six schools (three schools per district). Amount of fresh F/V taken, consumed, and wasted. Negative binomial multivariable regression examined placement of salad bars, adjusting for sex, grade, race/ethnicity, free/reduced status, day of the week, and nesting of students within schools. Almost all students (98.6%) in the schools with salad bars inside serving lines self-served F/V compared with only 22.6% of students in the schools with salad bars outside lines (adjusted prevalence ratio=5.38; 95% CI 4.04 to 7.17). Similarly, students at schools with salad bars inside the line had greater prevalence of consuming any F/V compared with students in schools with salad bars outside the line (adjusted prevalence ratio=4.83; 95% CI 3.40 to 6.81). On average, students with the salad bar outside the line wasted less F/V compared with those with salad bars inside the line (30% vs 48%, respectively). Few students visited salad bars located outside the lunch line. Salad bars inside the lunch line resulted in significantly greater fresh F/V taken, consumed, and wasted. When possible, schools should try to include salad bars inside the line to increase students' exposure to F/V. Copyright © 2016 Academy of Nutrition and Dietetics. Published by Elsevier Inc. All rights reserved.

First Discovery of Acetone Extract from Cottonseed Oil Sludge as a Novel Antiviral Agent against Plant Viruses

PubMed Central

Zhao, Lei; Feng, Chaohong; Hou, Caiting; Hu, Lingyun; Wang, Qiaochun; Wu, Yunfeng

2015-01-01

A novel acetone extract from cottonseed oil sludge was firstly discovered against plant viruses including Tobacco mosaic virus (TMV), Rice stripe virus (RSV) and Southern rice black streaked dwarf virus (SRBSDV). Gossypol and β-sitosterol separated from the acetone extract were tested for their effects on anti-TMV and analysed by nuclear magnetic resonance (NMR) assay. In vivo and field trials in different geographic distributions and different host varieties declared that this extract mixture was more efficient than the commercial agent Ningnanmycin with a broad spectrum of anti-plant-viruses activity. No phytotoxic activity was observed in the treated plants and environmental toxicology showed that this new acetone extract was environmentally friendly, indicating that this acetone extract has potential application in the control of plant virus in the future. PMID:25705894

First discovery of acetone extract from cottonseed oil sludge as a novel antiviral agent against plant viruses.

PubMed

Zhao, Lei; Feng, Chaohong; Hou, Caiting; Hu, Lingyun; Wang, Qiaochun; Wu, Yunfeng

2015-01-01

A novel acetone extract from cottonseed oil sludge was firstly discovered against plant viruses including Tobacco mosaic virus (TMV), Rice stripe virus (RSV) and Southern rice black streaked dwarf virus (SRBSDV). Gossypol and β-sitosterol separated from the acetone extract were tested for their effects on anti-TMV and analysed by nuclear magnetic resonance (NMR) assay. In vivo and field trials in different geographic distributions and different host varieties declared that this extract mixture was more efficient than the commercial agent Ningnanmycin with a broad spectrum of anti-plant-viruses activity. No phytotoxic activity was observed in the treated plants and environmental toxicology showed that this new acetone extract was environmentally friendly, indicating that this acetone extract has potential application in the control of plant virus in the future.

A Cosmetic Content-Based Nutrition Education Program Improves Fruit and Vegetable Consumption Among Grade 11 Thai Students.

PubMed

Somsri, Pattraporn; Satheannoppakao, Warapone; Tipayamongkholgul, Mathuros; Vatanasomboon, Paranee; Kasemsup, Rachada

2016-03-01

To examine and compare the effectiveness of a cosmetic content-based nutrition education (CCBNEd) program and a health content-based nutrition education (HCBNEd) program on the promotion of fruit and vegetable (F&V) consumption. Quasi-experimental. Three secondary schools in Nonthaburi, Thailand. Three classes of students were randomly assigned to 3 study groups: experimental group 1 (n = 41) participated in the CCBNEd program, experimental group 2 (n = 35) experienced the HCBNEd program, and a comparison group (n = 37) did not participate in a program. All groups received F&V information. Data were collected between July and September, 2013. Knowledge about F&V, attitude toward F&V consumption, and the amount and variety of F&V consumed were measured at baseline, posttest, and follow-up. Nonparametric statistics were used to compare the programs' effectiveness. After the test, experimental group 1 had significantly increased knowledge scores, attitude scores, and the amount and variety of F&V consumed compared with those at baseline (P < .001). These positive changes were maintained until follow-up. In experimental group 2, knowledge and attitude scores increased (P < .001) at posttest and then decreased at follow-up whereas the comparison group positively changed only in knowledge. The CCBNEd program was most effective at increasing F&V consumption. Copyright © 2016 Society for Nutrition Education and Behavior. Published by Elsevier Inc. All rights reserved.

Design of Embedded-Hybrid Antimicrobial Peptides with Enhanced Cell Selectivity and Anti-Biofilm Activity

PubMed Central

Xu, Wei; Zhu, Xin; Tan, Tingting; Li, Weizhong; Shan, Anshan

2014-01-01

Antimicrobial peptides have attracted considerable attention because of their broad-spectrum antimicrobial activity and their low prognostic to induce antibiotic resistance which is the most common source of failure in bacterial infection treatment along with biofilms. The method to design hybrid peptide integrating different functional domains of peptides has many advantages. In this study, we designed an embedded-hybrid peptide R-FV-I16 by replacing a functional defective sequence RR7 with the anti-biofilm sequence FV7 embedded in the middle position of peptide RI16. The results demonstrated that the synthetic hybrid the peptide R-FV-I16 had potent antimicrobial activity over a wide range of Gram-negative and Gram-positive bacteria, as well as anti-biofilm activity. More importantly, R-FV-I16 showed lower hemolytic activity and cytotoxicity. Fluorescent assays demonstrated that R-FV-I16 depolarized the outer and the inner bacterial membranes, while scanning electron microscopy and transmission electron microscopy further indicated that this peptide killed bacterial cells by disrupting the cell membrane, thereby damaging membrane integrity. Results from SEM also provided evidence that R-FV-I16 inherited anti-biofilm activity from the functional peptide sequence FV7. Embedded-hybrid peptides could provide a new pattern for combining different functional domains and showing an effective avenue to screen for novel antimicrobial agents. PMID:24945359

Reaching Low-Income Mothers to Improve Family Fruit and Vegetable Intake: Food Hero Social Marketing Campaign—Research Steps, Development and Testing

PubMed Central

Tobey, Lauren N.; Koenig, Harold F.; Brown, Nicole A.; Manore, Melinda M.

2016-01-01

The objective of this study was to create/test a social marketing campaign to increase fruit/vegetable (FV) intake within Oregon Supplemental Nutrition Assistance Program (SNAP) eligible families. Focus groups (n = 2) and pre/post campaign phone surveys (n = 2082) were conducted in intervention counties (IC) and one control county. Participants were female (86%–100%) with 1–2 children at home. Mean FV intake/without juice was 3.1 servings/day; >50% preferred the Internet for delivery of healthy eating information. Participants reported time/financial burdens, low household FV variety and desirability of frozen/canned FV, and acceptance of positive messages. A Food Hero (FH) campaign was created/delivered daily August–October 2009 to mothers through multiple channels (e.g., grocery stores, online, educators). Results showed that the IC had better FH name recall (12%) and interpretation of intended messages (60%) vs. control (3%, 23%, respectively). Compared to controls, the IC were less likely to report healthy food preparation as time consuming or a FV rich diet expensive, and it was easier to get their family to eat fruit. Results did not vary based on county/household characteristics. The FH campaign increased FH awareness and positive FV beliefs. A longer campaign with FV assessments will increase understanding of the target audience, and allow for campaign refinement. PMID:27649233

Identification of Fusarium virguliforme FvTox1-Interacting Synthetic Peptides for Enhancing Foliar Sudden Death Syndrome Resistance in Soybean

PubMed Central

Wang, Bing; Swaminathan, Sivakumar; Bhattacharyya, Madan K.

2015-01-01

Soybean is one of the most important crops grown across the globe. In the United States, approximately 15% of the soybean yield is suppressed due to various pathogen and pests attack. Sudden death syndrome (SDS) is an emerging fungal disease caused by Fusarium virguliforme. Although growing SDS resistant soybean cultivars has been the main method of controlling this disease, SDS resistance is partial and controlled by a large number of quantitative trait loci (QTL). A proteinacious toxin, FvTox1, produced by the pathogen, causes foliar SDS. Earlier, we demonstrated that expression of an anti-FvTox1 single chain variable fragment antibody resulted in reduced foliar SDS development in transgenic soybean plants. Here, we investigated if synthetic FvTox1-interacting peptides, displayed on M13 phage particles, can be identified for enhancing foliar SDS resistance in soybean. We screened three phage-display peptide libraries and discovered four classes of M13 phage clones displaying FvTox1-interacting peptides. In vitro pull-down assays and in vivo interaction assays in yeast were conducted to confirm the interaction of FvTox1 with these four synthetic peptides and their fusion-combinations. One of these peptides was able to partially neutralize the toxic effect of FvTox1 in vitro. Possible application of the synthetic peptides in engineering SDS resistance soybean cultivars is discussed. PMID:26709700

Successful construction and stable expression of an anti-CD45RA scFv-EGFP fusion protein in Chinese hamster ovary cells.

PubMed

Wang, Zhujun; Chen, Yuanyuan; Li, Sisi; Cheng, Yuping; Zhao, Haizhao; Jia, Ming; Luo, Zebin; Tang, Yongmin

2014-02-01

CD45RA has been found highly expressed on leukemia cells and may be a potential target of the disease. In this study, an anti-CD45RA single-chain antibody fragment (scFv3A4) was genetically linked to the N terminus of the enhanced green fluorescent protein (EGFP) to generate a scFv3A4-EGFP fusion protein. The scFv3A4-EGFP with a molecular weight of 57kDa was stably expressed and secreted from the transfected CHO cells through the ER/Golgi-dependent pathway. The fusion protein was soluble in the culture supernatant and the yield was 1350μg/L. Flow cytometry analysis showed that the scFv3A4-EGFP had the same binding site and a very similar reactivity pattern with its parental murine monoclonal antibody (mAb) 3A4. Furthermore, comparing to conventional labeled 3A4-FITC antibody, the scFv3A4-EGFP was more resistant to illumination and more suitable for immunofluorescence histology (IFH) detection. Therefore, the scFv3A4-EGFP fusion protein can be a powerful tool to investigate the targeting of CD45RA on leukemia cells, biological activity of the target and possibly for the genetic manipulation of the antibody. Copyright © 2013 Elsevier Inc. All rights reserved.

Reaching Low-Income Mothers to Improve Family Fruit and Vegetable Intake: Food Hero Social Marketing Campaign-Research Steps, Development and Testing.

PubMed

Tobey, Lauren N; Koenig, Harold F; Brown, Nicole A; Manore, Melinda M

2016-09-13

The objective of this study was to create/test a social marketing campaign to increase fruit/vegetable (FV) intake within Oregon Supplemental Nutrition Assistance Program (SNAP) eligible families. Focus groups (n = 2) and pre/post campaign phone surveys (n = 2082) were conducted in intervention counties (IC) and one control county. Participants were female (86%-100%) with 1-2 children at home. Mean FV intake/without juice was 3.1 servings/day; >50% preferred the Internet for delivery of healthy eating information. Participants reported time/financial burdens, low household FV variety and desirability of frozen/canned FV, and acceptance of positive messages. A Food Hero (FH) campaign was created/delivered daily August-October 2009 to mothers through multiple channels (e.g., grocery stores, online, educators). Results showed that the IC had better FH name recall (12%) and interpretation of intended messages (60%) vs. control (3%, 23%, respectively). Compared to controls, the IC were less likely to report healthy food preparation as time consuming or a FV rich diet expensive, and it was easier to get their family to eat fruit. Results did not vary based on county/household characteristics. The FH campaign increased FH awareness and positive FV beliefs. A longer campaign with FV assessments will increase understanding of the target audience, and allow for campaign refinement.

Administrative and School Nutrition Perspectives of Salad Bar Operations in Public Schools

ERIC Educational Resources Information Center

Andersen, Lori; Myers, Leann; O'Malley, Keelia; Rose, Donald; Johnson, Carolyn C.

2016-01-01

Purposes/Objectives: Fruit and vegetable (F/V) consumption may aid in childhood obesity prevention. F/V consumption in youth is low. School-based salad bars (SBs) may improve F/V access in youth. The purpose of this study was to explore administrative and school nutrition personnel perspectives related to adoption and continued implementation of…

Cross-Country comparison of professionals' perceptions of the effectiveness of parenting practices to promote fruit and vegetable consumption in preschool children

USDA-ARS?s Scientific Manuscript database

Obesity prevention often includes promotion of fruit and vegetable (FV) intake. Parents may be an important influence on FV intake for preschool children. Which parenting practices (PP) are effective in getting children to eat FV throughout childhood is unclear. A national variations in professional...

Measurement of Factor V Activity in Human Plasma Using a Microplate Coagulation Assay

PubMed Central

Tilley, Derek; Levit, Irina; Samis, John A.

2012-01-01

In response to injury, blood coagulation is activated and results in generation of the clotting protease, thrombin. Thrombin cleaves fibrinogen to fibrin which forms an insoluble clot that stops hemorrhage. Factor V (FV) in its activated form, FVa, is a critical cofactor for the protease FXa and accelerator of thrombin generation during fibrin clot formation as part of prothrombinase 1, 2. Manual FV assays have been described 3, 4, but they are time consuming and subjective. Automated FV assays have been reported 5-7, but the analyzer and reagents are expensive and generally provide only the clot time, not the rate and extent of fibrin formation. The microplate platform is preferred for measuring enzyme-catalyzed events because of convenience, time, cost, small volume, continuous monitoring, and high-throughput 8, 9. Microplate assays have been reported for clot lysis 10, platelet aggregation 11, and coagulation Factors 12, but not for FV activity in human plasma. The goal of the method was to develop a microplate assay that measures FV activity during fibrin formation in human plasma. This novel microplate method outlines a simple, inexpensive, and rapid assay of FV activity in human plasma. The assay utilizes a kinetic microplate reader to monitor the absorbance change at 405nm during fibrin formation in human plasma (Figure 1) 13. The assay accurately measures the time, initial rate, and extent of fibrin clot formation. It requires only μl quantities of plasma, is complete in 6 min, has high-throughput, is sensitive to 24-80pM FV, and measures the amount of unintentionally activated (1-stage activity) and thrombin-activated FV (2-stage activity) to obtain a complete assessment of its total functional activity (2-stage activity - 1-stage activity). Disseminated intravascular coagulation (DIC) is an acquired coagulopathy that most often develops from pre-existing infections 14. DIC is associated with a poor prognosis and increases mortality above the pre-existing pathology 15. The assay was used to show that in 9 patients with DIC, the FV 1-stage, 2-stage, and total activities were decreased, on average, by 54%, 44%, and 42%, respectively, compared with normal pooled human reference plasma (NHP). The FV microplate assay is easily adaptable to measure the activity of any coagulation factor. This assay will increase our understanding of FV biochemistry through a more accurate and complete measurement of its activity in research and clinical settings. This information will positively impact healthcare environments through earlier diagnosis and development of more effective treatments for coagulation disorders, such as DIC. PMID:22987015

Measurement of factor v activity in human plasma using a microplate coagulation assay.

PubMed

Tilley, Derek; Levit, Irina; Samis, John A

2012-09-09

In response to injury, blood coagulation is activated and results in generation of the clotting protease, thrombin. Thrombin cleaves fibrinogen to fibrin which forms an insoluble clot that stops hemorrhage. Factor V (FV) in its activated form, FVa, is a critical cofactor for the protease FXa and accelerator of thrombin generation during fibrin clot formation as part of prothrombinase (1, 2). Manual FV assays have been described (3, 4), but they are time consuming and subjective. Automated FV assays have been reported (5-7), but the analyzer and reagents are expensive and generally provide only the clot time, not the rate and extent of fibrin formation. The microplate platform is preferred for measuring enzyme-catalyzed events because of convenience, time, cost, small volume, continuous monitoring, and high-throughput (8, 9). Microplate assays have been reported for clot lysis (10), platelet aggregation (11), and coagulation Factors (12), but not for FV activity in human plasma. The goal of the method was to develop a microplate assay that measures FV activity during fibrin formation in human plasma. This novel microplate method outlines a simple, inexpensive, and rapid assay of FV activity in human plasma. The assay utilizes a kinetic microplate reader to monitor the absorbance change at 405 nm during fibrin formation in human plasma (Figure 1) (13). The assay accurately measures the time, initial rate, and extent of fibrin clot formation. It requires only μl quantities of plasma, is complete in 6 min, has high-throughput, is sensitive to 24-80 pM FV, and measures the amount of unintentionally activated (1-stage activity) and thrombin-activated FV (2-stage activity) to obtain a complete assessment of its total functional activity (2-stage activity - 1-stage activity). Disseminated intravascular coagulation (DIC) is an acquired coagulopathy that most often develops from pre-existing infections (14). DIC is associated with a poor prognosis and increases mortality above the pre-existing pathology (15). The assay was used to show that in 9 patients with DIC, the FV 1-stage, 2-stage, and total activities were decreased, on average, by 54%, 44%, and 42%, respectively, compared with normal pooled human reference plasma (NHP). The FV microplate assay is easily adaptable to measure the activity of any coagulation factor. This assay will increase our understanding of FV biochemistry through a more accurate and complete measurement of its activity in research and clinical settings. This information will positively impact healthcare environments through earlier diagnosis and development of more effective treatments for coagulation disorders, such as DIC.

Force-velocity properties' contribution to bilateral deficit during ballistic push-off.

PubMed

Samozino, Pierre; Rejc, Enrico; di Prampero, Pietro Enrico; Belli, Alain; Morin, Jean-Benoît

2014-01-01

The objective of this study is to quantify the contribution of the force-velocity (F-v) properties to bilateral force deficit (BLD) in ballistic lower limb push-off and to relate it to individual F-v mechanical properties of the lower limbs. The F-v relation was individually assessed from mechanical measurements for 14 subjects during maximal ballistic lower limb push-offs; its contribution to BLD was then investigated using a theoretical macroscopic approach, considering both the mechanical constraints of movement dynamics and the maximal external capabilities of the lower limb neuromuscular system. During ballistic lower limb push-off, the maximum force each lower limb can produce was lower during bilateral than unilateral actions, thus leading to a BLD of 36.7% ± 5.7%. The decrease in force due to the F-v mechanical properties amounted to 19.9% ± 3.6% of the force developed during BL push-offs, which represents a nonneural contribution to BLD of 43.5% ± 9.1%. This contribution to BLD that cannot be attributed to changes in neural features was negatively correlated to the maximum unloaded extension velocity of the lower limb (r = -0.977, P < 0.001). During ballistic lower limb push-off, BLD is due to both neural alterations and F-v mechanical properties, the latter being associated with the change in movement velocity between bilateral and unilateral actions. The level of the contribution of the F-v properties depends on the individual F-v mechanical profile of the entire lower limb neuromuscular system: the more the F-v profile is oriented toward velocity capabilities, the lower the loss of force from unilateral to bilateral push-offs due to changes in movement velocity.

Interrelationship between food security status, home availability of variety of fruits and vegetables and their dietary intake among low-income pregnant women.

PubMed

Nunnery, Danielle L; Labban, Jeffrey D; Dharod, Jigna M

2018-03-01

To (i) determine differences in the availability of variety of fruits and vegetables (F&V) at home by food security status; and (ii) examine the inter-associations between food security status, availability of variety of F&V at home and frequency of F&V intake, among low-income pregnant women. Design/Setting Participants were interviewed to collect food security status, home availability of variety of F&V and frequency of F&V intake. Bivariate analyses, multivariate regression and exploratory mediation analyses were conducted using IBM SPSS Statistics version 23.0 and the PROCESS macro. Low-income pregnant women (n 198) were interviewed if they were ≥18 years of age, in the second trimester of pregnancy, and spoke English or Spanish. Low/very low food security was found among 43 % of participants. Compared with fully food-secure participants, very low food-secure participants reported a lower variety of fruits (P=0·028) and vegetables (P=0·058) available at home. Mediation analyses indicated that through home availability of variety of fresh F&V, food security status was associated with the daily intake of F&V (indirect effect (95 % CI): fresh fruits, -0·039 (-0·074, -0·013); fresh vegetables, -0·048 (-0·083, -0·023)). As food security worsened, the available variety of fresh F&V decreased, which was associated with lower intake. The study highlights the interlink between food security, home food environment and diet quality, and the importance of nutrition education intervention to promote a healthy home food environment and improved pregnancy outcomes among low-income women.

The WRKY transcription factors in the diploid woodland strawberry Fragaria vesca: Identification and expression analysis under biotic and abiotic stresses.

PubMed

Wei, Wei; Hu, Yang; Han, Yong-Tao; Zhang, Kai; Zhao, Feng-Li; Feng, Jia-Yue

2016-08-01

WRKY proteins comprise a large family of transcription factors that play important roles in response to biotic and abiotic stresses and in plant growth and development. To date, little is known about the WRKY gene family in strawberry. In this study, we identified 62 WRKY genes (FvWRKYs) in the wild diploid woodland strawberry (Fragaria vesca, 2n = 2x = 14) accession Heilongjiang-3. According to the phylogenetic analysis and structural features, these identified strawberry FvWRKY genes were classified into three main groups. In addition, eight FvWRKY-GFP fusion proteins showed distinct subcellular localizations in Arabidopsis mesophyll protoplasts. Furthermore, we examined the expression of the 62 FvWRKY genes in 'Heilongjiang-3' under various conditions, including biotic stress (Podosphaera aphanis), abiotic stresses (drought, salt, cold, and heat), and hormone treatments (abscisic acid, ethephon, methyl jasmonate, and salicylic acid). The expression levels of 33 FvWRKY genes were upregulated, while 12 FvWRKY genes were downregulated during powdery mildew infection. FvWRKY genes responded to drought and salt treatment to a greater extent than to temperature stress. Expression profiles derived from quantitative real-time PCR suggested that 11 FvWRKY genes responded dramatically to various stimuli at the transcriptional level, indicating versatile roles in responses to biotic and abiotic stresses. Interaction networks revealed that the crucial pathways controlled by WRKY proteins may be involved in the differential response to biotic stress. Taken together, the present work may provide the basis for future studies of the genetic modification of WRKY genes for pathogen resistance and stress tolerance in strawberry. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

High efficient expression of a functional humanized single-chain variable fragment (scFv) antibody against CD22 in Pichia pastoris.

PubMed

Zarei, Najmeh; Vaziri, Behrouz; Shokrgozar, Mohammad Ali; Mahdian, Reza; Fazel, Ramin; Khalaj, Vahid

2014-12-01

Single-chain variable fragments (scFvs) have recently emerged as attractive candidates in targeted immunotherapy of various malignancies. The anti-CD22 scFv is able to target CD22, on B cell surface and is being considered as a promising molecule in targeted immunotherapy of B cell malignancies. The recombinant anti-CD22 scFv has been successfully expressed in Escherichia coli; however, the insufficient production yield has been a major bottleneck for its therapeutic application. The methylotrophic yeast Pichia pastoris has become a highly popular expression host for the production of a wide variety of recombinant proteins such as antibody fragments. In this study, we used the Pichia expression system to express a humanized scFv antibody against CD22. The full-length humanized scFv gene was codon optimized, cloned into the pPICZαA and expressed in GS115 strain. The maximum production level of the scFv (25 mg/L) were achieved at methanol concentration, 1 %; pH 6.0; inoculum density, OD600 = 3 and the induction time of 72 h. The correlation between scFv gene dosage and expression level was also investigated by real-time PCR, and the results confirmed the presence of such correlation up to five gene copies. Immunofluorescence and flow cytometry studies and Biacore analysis demonstrated binding to CD22 on the surface of human lymphoid cell line Raji and recombinant soluble CD22, respectively. Taken together, the presented data suggest that the Pichia pastoris can be considered as an efficient host for the large-scale production of anti-CD22 scFv as a promising carrier for targeted drug delivery in treatment of CD22(+) B cell malignancies.

Evaluation of the impact of school gardening interventions on children's knowledge of and attitudes towards fruit and vegetables. A cluster randomised controlled trial.

PubMed

Hutchinson, Jayne; Christian, Meaghan Sarah; Evans, Charlotte Elizabeth Louise; Nykjaer, Camilla; Hancock, Neil; Cade, Janet Elizabeth

2015-08-01

Involvement of children in gardening has the potential to increase liking of fruit and vegetables (FV) and consequently, intake, but research results are mixed. School gardening led by external specialists such as the Royal Horticultural Society (RHS) could have more impact than teacher-led gardening on children's knowledge of, and attitudes towards, FV. Data from a cluster randomised controlled trial were used to compare a RHS-led school gardening intervention with a teacher-led gardening intervention amongst 7-10 year olds in 21 London schools. A short questionnaire was developed and used to identify children's knowledge and attitudes towards FV consumption before the garden intervention and 18 months afterwards. Results from multilevel regression models, both unadjusted and adjusted for baseline responses and socio-demographic factors, were reported. Attitudes to FV intake were compared between groups. Change in FV knowledge was used to predict change in FV consumption assessed using 24-hour food diaries. In comparison with the RHS-led group (n = 373), teacher-led children (n = 404) were more likely to agree they ate lots of fruit (p < 0.009) and tried new fruits (p = 0.045), but RHS-led gardening was associated with a greater increase in the total number of vegetables recognised (p = 0.031). No other differences in improvements in attitudes, or associations between change in FV recognition and intake were found. In relation to improvements in children's recognition and attitudes towards eating FV, this trial produced limited evidence that gardening activity packages led by external specialists (RHS-led) provide additional benefits over those led by teachers trained by the RHS. Indeed, the latter were potentially more effective. Copyright © 2015 Elsevier Ltd. All rights reserved.

A multi-level intervention in worksites to increase fruit and vegetable access and intake: Rationale, design and methods of the 'Good to Go' cluster randomized trial.

PubMed

Risica, Patricia M; Gorham, Gemma; Dionne, Laura; Nardi, William; Ng, Doug; Middler, Reese; Mello, Jennifer; Akpolat, Rahmet; Gettens, Katelyn; Gans, Kim M

2018-02-01

Fruit and vegetable (F&V) consumption is an important contributor to chronic disease prevention. However, most Americans do not eat adequate amounts. The worksite is an advantageous setting to reach large, diverse segments of the population with interventions to increase F&V intake, but research gaps exist. No studies have evaluated the implementation of mobile F&V markets at worksites nor compared the effectiveness of such markets with or without nutrition education. This paper describes the protocol for Good to Go (GTG), a cluster randomized trial to evaluate F&V intake change in employees from worksites randomized into three experimental arms: discount, fresh F&V markets (Access Only arm); markets plus educational components including campaigns, cooking demonstrations, videos, newsletters, and a web site (Access Plus arm); and an attention placebo comparison intervention on physical activity and stress reduction (Comparison). Secondary aims include: 1) Process evaluation to determine costs, reach, fidelity, and dose as well as the relationship of these variables with changes in F&V intake; 2) Applying a mediating variable framework to examine relationships of psychosocial factors/determinants with changes in F&V consumption; and 3) Cost effectiveness analysis of the different intervention arms. The GTG study will fill important research gaps in the field by implementing a rigorous cluster randomized trial to evaluate the efficacy of an innovative environmental intervention providing access and availability to F&V at the worksite and whether this access intervention is further enhanced by accompanying educational interventions. GTG will provide an important contribution to public health research and practice. Trial registration number NCT02729675, ClinicalTrials.gov. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Genetic modification of mesenchymal stem cells to express a single-chain antibody against EGFRvIII on the cell surface.

PubMed

Balyasnikova, Irina V; Franco-Gou, Rosa; Mathis, J Michael; Lesniak, Maciej S

2010-06-01

Human adult mesenchymal stem cells (hMSCs) are under active investigation as cellular carriers for gene therapy. hMSCs possess natural tropism toward tumours; however, the targeting of hMSCs to specific cell populations within tumours is unexplored. In the case of glioblastoma multiforme (GBM), at least half of the tumours express EGFRvIII on the cell surface, an ideal target for antibody-mediated gene/drug delivery. In this study, we investigated the feasibility of genetically modifying hMSCs to express a single-chain antibody (scFv) to EGFRvIII on their surfaces. Nucleofection was used to transfect hMSCs with cDNA encoding scFv EGFRvIII fused with PDGFR or human B7-1 transmembrane domains. The expression of scFv EGFRvIII on the cell surface was assessed by FACS. A stable population of scFv EGFRvIII-expressing hMSCs was selected, based on antibiotic resistance, and enriched using FACS. We found that nucleofection allows the efficient expression of scFv EGFRvIII on the cell surface of hMSCs. hMSCs transfected with the construct encoding scFv EGFRvIII as a fusion with PDGFRtm showed scFv EGFRvIII expression in up to 86% of cells. Most importantly, human MSCs expressing scFv against EGFRvIII demonstrated enhanced binding to U87-EGFRvIII cells in vitro and significantly increased retention in human U87-EGFRvIII-expressing tumours in vivo. In summary, we provide the first conclusive evidence of genetic modification of hMSCs with a single-chain antibody against an antigen expressed on the surface of tumour cells, thereby opening up a new venue for enhanced delivery of gene therapy applications in the context of malignant brain cancer. Copyright 2009 John Wiley & Sons, Ltd.

Play with your food! Sensory play is associated with tasting of fruits and vegetables in preschool children.

PubMed

Coulthard, Helen; Sealy, Annemarie

2017-06-01

The objective of the current study was to ascertain whether taking part in a sensory play activity with real fruits and vegetables (FV) can encourage tasting in preschool children, compared to a non-food activity or visual exposure to the activity. Three to four year old pre-school children (N = 62) were recruited from three preschool nursery classes from a school in Northamptonshire, UK. A between participants experimental study was conducted with each class assigned to one of three conditions; sensory FV play, sensory non-food play and visual FV exposure. Parental report of several baseline variables were taken; child baseline liking of the foods used in the study, parental and child FV consumption (portions/day), child neophobia and child tactile sensitivity. Outcome measures were the number of fruits and vegetables tasted in a post experiment taste test which featured (n = 5) or did not feature (n = 3) in the task. Analyses of covariance controlling for food neophobia and baseline liking of foods, showed that after the activity children in the sensory FV play condition tried more FV than both children in the non-food sensory play task (p < 0.001) and children in the visual FV exposure task (p < 0.001). This was true not only for five foods used in the activity (p < 0.001), but also three foods that were not used in the activity (p < 0.05). Sensory play activities using fruits and vegetables may encourage FV tasting in preschool children more than non food play or visual exposure alone. Long term intervention studies need to be carried out to see if these effects can be sustained over time. Copyright © 2017 Elsevier Ltd. All rights reserved.

A multi-level intervention in worksites to increase fruit and vegetable access and intake: Rationale, design and methods of the ‘Good to Go’ cluster randomized trial

PubMed Central

Risica, Patricia M.; Gorham, Gemma; Dionne, Laura; Nardi, William; Ng, Doug; Middler, Reese; Mello, Jennifer; Akpolat, Rahmet; Gettens, Katelyn; Gans, Kim M.

2018-01-01

Background Fruit and vegetable (F&V) consumption is an important contributor to chronic disease prevention. However, most Americans do not eat adequate amounts. The worksite is an advantageous setting to reach large, diverse segments of the population with interventions to increase F&V intake, but research gaps exist. No studies have evaluated the implementation of mobile F&V markets at worksites nor compared the effectiveness of such markets with or without nutrition education. Methods This paper describes the protocol for Good to Go (GTG), a cluster randomized trial to evaluate F&V intake change in employees from worksites randomized into three experimental arms: discount, fresh F&V markets (Access Only arm); markets plus educational components including campaigns, cooking demonstrations, videos, newsletters, and a web site (Access Plus arm); and an attention placebo comparison intervention on physical activity and stress reduction (Comparison). Secondary aims include: 1) Process evaluation to determine costs, reach, fidelity, and dose as well as the relationship of these variables with changes in F&V intake; 2) Applying a mediating variable framework to examine relationships of psychosocial factors/determinants with changes in F&V consumption; and 3) Cost effectiveness analysis of the different intervention arms. Discussion The GTG study will fill important research gaps in the field by implementing a rigorous cluster randomized trial to evaluate the efficacy of an innovative environmental intervention providing access and availability to F&V at the worksite and whether this access intervention is further enhanced by accompanying educational interventions. GTG will provide an important contribution to public health research and practice. Trial registration number NCT02729675, ClinicalTrials.gov PMID:29242108

An efficient strategy for cell-based antibody library selection using an integrated vector system.

PubMed

Yoon, Hyerim; Song, Jin Myung; Ryu, Chun Jeih; Kim, Yeon-Gu; Lee, Eun Kyo; Kang, Sunghyun; Kim, Sang Jick

2012-09-18

Cell panning of phage-displayed antibody library is a powerful tool for the development of therapeutic and imaging agents since disease-related cell surface proteins in native complex conformation can be directly targeted. Here, we employed a strategy taking advantage of an integrated vector system which allows rapid conversion of scFv-displaying phage into scFv-Fc format for efficient cell-based scFv library selection on a tetraspanin protein, CD9. A mouse scFv library constructed by using a phagemid vector, pDR-D1 was subjected to cell panning against stable CD9 transfectant, and the scFv repertoire from the enriched phage pool was directly transferred to a mammalian cassette vector, pDR-OriP-Fc1. The resulting constructs enabled transient expression of enough amounts of scFv-Fcs in HEK293E cells, and flow cytometric screening of binders for CD9 transfectant could be performed simply by using the culture supernatants. All three clones selected from the screening showed correct CD9-specificity. They could immunoprecipitate CD9 molecules out of the transfectant cell lysate and correctly stain endogenous CD9 expression on cancer cell membrane. Furthermore, competition assay with a known anti-CD9 monoclonal antibody (mAb) suggested that the binding epitopes of some of them overlap with that of the mAb which resides within the large extracellular loop of CD9. This study demonstrates that scFv-Fc from mammalian transient expression can be chosen as a reliable format for rapid screening and validation in cell-based scFv library selection, and the strategy described here will be applicable to efficient discovery of antibodies to diverse cell-surface targets.

The influence of family violence and child marriage on unmet need for family planning in Jordan.

PubMed

Clark, Cari Jo; Spencer, Rachael A; Khalaf, Inaam A; Gilbert, Louisa; El-Bassel, Nabila; Silverman, Jay G; Raj, Anita

2017-04-01

Risk for unmet need for contraception is associated with men's perpetration of intimate partner violence (IPV) against women and may be influenced by violence perpetrated by other family members (family violence, FV). Women who married as minors may be most vulnerable to the potential compounding effect of IPV and FV on unmet need. Using nationally representative data from the 2012 Jordan Population and Family Health Survey we examined unmet need by exposure to IPV and FV by women's age at marriage (<18, 18+ years). Logistic regression was used to test whether IPV and FV were independently associated with unmet need, by age at marriage. Interaction terms (IPV×FV) were tested in both models. Stratification by FV was employed to clarify the interpretation of significant interactions. IPV increased the odds of unmet need by 87% [adjusted odds ratio (AOR) 1.87; 95% confidence interval (95% CI) 1.13-3.10] and 76% (AOR 1.76; 95% CI 1.30-2.38) among women who married prior to and after the age of 18 years, respectively. Women married as minors who experienced IPV and FV had a four-fold higher likelihood of having an unmet need (AOR 6.75; 95% CI 1.95-23.29) compared to those experiencing only IPV (AOR 1.49; 95% CI 0.84-2.38). No interaction between IPV and FV was detected for women married at or above majority. Laws that prohibit child marriage should be strengthened and health sector screening for violence experience could help identify women at risk of unmet need and improve women's reproductive agency. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

Fruit and vegetable availability and selection: federal food package revisions, 2009.

PubMed

Zenk, Shannon N; Odoms-Young, Angela; Powell, Lisa M; Campbell, Richard T; Block, Daniel; Chavez, Noel; Krauss, Ramona C; Strode, Steven; Armbruster, James

2012-10-01

With nearly 49,000 authorized retailers nationwide, a policy change that added fruits and vegetables (FV) to the U.S. Department of Agriculture's Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) food packages in 2009 had the potential to expand neighborhood FV availability. This study examined changes in availability and selection of commonly consumed and culturally specific FV at authorized retailers (WIC vendors) before and after implementation of the revised WIC food packages. Quasi-experimental, one-group design with two pre-policy observations and one post-policy observation. Trained observers assessed a list of fresh, frozen, and canned FV at each vendor in seven northern Illinois counties. Eight indices of FV availability and selection were derived. Multiple regression estimated relationships. Data were collected in 2008-2010 and analyzed in 2011. Overall, availability and selection of commonly consumed fresh FV and availability of African-American culturally specific fresh FV improved after implementation of the new policy. Modest improvements in the overall availability of canned low-sodium vegetables and frozen FV were observed. Changes differed by vendor type (large vendor, small vendor, and pharmacy). Changes in availability or selection did not differ by neighborhood characteristics (population density, median household income, racial/ethnic composition). Expansion of WIC foods was associated with small positive externalities on the food environment. Larger subsidies to create more demand and more-substantial stocking requirements for retailers may yield significantly larger improvements and thus warrant further investigation. Approaches targeting rural, low-income, and racial/ethnic minority neighborhoods also may be needed. Copyright © 2012 American Journal of Preventive Medicine. Published by Elsevier Inc. All rights reserved.

Wheat cultivars selected for high Fv /Fm under heat stress maintain high photosynthesis, total chlorophyll, stomatal conductance, transpiration and dry matter.

PubMed

Sharma, Dew Kumari; Andersen, Sven Bode; Ottosen, Carl-Otto; Rosenqvist, Eva

2015-02-01

The chlorophyll fluorescence parameter Fv /Fm reflects the maximum quantum efficiency of photosystem II (PSII) photochemistry and has been widely used for early stress detection in plants. Previously, we have used a three-tiered approach of phenotyping by Fv /Fm to identify naturally existing genetic variation for tolerance to severe heat stress (3 days at 40°C in controlled conditions) in wheat (Triticum aestivum L.). Here we investigated the performance of the previously selected cultivars (high and low group based on Fv /Fm value) in terms of growth and photosynthetic traits under moderate heat stress (1 week at 36/30°C day/night temperature in greenhouse) closer to natural heat waves in North-Western Europe. Dry matter accumulation after 7 days of heat stress was positively correlated to Fv /Fm . The high Fv /Fm group maintained significantly higher total chlorophyll and net photosynthetic rate (PN ) than the low group, accompanied by higher stomatal conductance (gs ), transpiration rate (E) and evaporative cooling of the leaf (ΔT). The difference in PN between the groups was not caused by differences in PSII capacity or gs as the variation in Fv /Fm and intracellular CO2 (Ci ) was non-significant under the given heat stress. This study validated that our three-tiered approach of phenotyping by Fv /Fm performed under increasing severity of heat was successful in identifying wheat cultivars differing in photosynthesis under moderate and agronomically more relevant heat stress. The identified cultivars may serve as a valuable resource for further studies to understand the physiological mechanisms underlying the genetic variability in heat sensitivity of photosynthesis. © 2014 Scandinavian Plant Physiology Society.

Radioiodination and biodistribution of the monoclonal antibody TU-20 and its scFv fragment

NASA Astrophysics Data System (ADS)

Kubaštová, H.; Kleinova, V.; Seifert, D.; Fišer, M.; Kranda, K.

2006-01-01

The ability of the monoclonal antibody TU-20 and its scFv fragment to specifically bind to the C-end of the class III beta-tubulin makes these preparations useful as potential diagnostics for in vivo determination of neurodegenerative diseases that entail degradation of neuronal cytoskeleton. To examine this hypothesis, TU-20 and its scFv were labelled with 125I and their properties were extensively investigated. TU-20 and its scFv were labelled via chloramine-T with the yield 90 95% and 64 78%, respectively. Their quality control, performed by an ELISA and gel electrophoresis, determined adequate properties for further studies. The in vitro experiment, involving autoradiography and immunohistochemistry of mice’ brain slices, enabled confirmation of preserved immunospecificity of the radiolabelled substances. Finally, the in vivo biodistribution proved differences in elimination of either TU-20, scFv TU-20, or iodide from the mice.

Methods of preparing and using single chain anti-tumor antibodies

DOEpatents

Cheung, Nai-Kong; Guo, Hong-Fen

2010-02-23

This invention provides a method for identifying cells expressing a target single chain antibody (scFv) directed against a target antigen from a collection of cells that includes cells that do not express the target scFv, comprising the step of combining the collection of cells with an anti-idiotype directed to an antibody specific for the target antigen and detecting interaction, if any, of the anti-idiotype with the cells, wherein the occurrence of an interaction identifies the cell as one which expresses the target scFv. This invention also provides a method for making a single chain antibody (scFv) directed against an antigen, wherein the selection of clones is made based upon interaction of those clones with an appropriate anti-idiotype, and heretofore inaccessible scFv so made. This invention provides the above methods or any combination thereof. Finally, this invention provides various uses of these methods.

Method for preparation of single chain antibodies

DOEpatents

Cheung, Nai-Kong V [New York, NY; Guo, Hong-fen [New York, NY

2012-04-03

This invention provides a method for identifying cells expressing a target single chain antibody (scFv) directed against a target antigen from a collection of cells that includes cells that do not express the target scFv, comprising the step of combining the collection of cells with an anti-idiotype directed to an antibody specific for the target antigen and detecting interaction, if any, of the anti-idiotype with the cells, wherein the occurrence of an interaction identifies the cell as one which expresses the target scFv. This invention also provides a method for making a single chain antibody (scFv) directed against an antigen, wherein the selection of clones is made based upon interaction of those clones with an appropriate anti-idiotype, and heretofore inaccessible scFv so made. This invention provides the above methods or any combination thereof. Finally, this invention provides various uses of these methods.

Role of vegetables and fruits in Mediterranean diets to prevent hypertension.

PubMed

Nuñez-Cordoba, J M; Alonso, A; Beunza, J J; Palma, S; Gomez-Gracia, E; Martinez-Gonzalez, M A

2009-05-01

Several studies support the effectiveness of increasing the consumption of fruits and vegetables (F&V) to prevent hypertension. However, none of them have been conducted in a Mediterranean setting. The aim of this study was to assess the association between F&V consumption and the risk of hypertension. A prospective Mediterranean study (the SUN cohort), including 8594 participants aged 20-95 years (mean, 41.1) with median follow-up of 49 months. Analyses according to the joint classification by olive oil and F&V consumption showed a significant inverse relation between F&V consumption and the risk of hypertension only among participants with a low olive oil consumption (<15 g per day). Also, tests for trend were significant only in the low olive oil intake stratum. We found a statistically significant interaction (P=0.01) between olive oil intake and F&V consumption. These data suggest a sub-additive effect of both food items.

The effect of simulated field storage conditions on the accuracy of rapid user-friendly blood pathogen detection kits.

PubMed

Bienek, Diane R; Charlton, David G

2012-05-01

Being able to test for the presence of blood pathogens at forward locations could reduce morbidity and mortality in the field. Rapid, user-friendly blood typing kits for detecting Human Immunodeficiency Virus (HIV), Hepatitis C Virus (HCV), and Hepatitis B Virus (HBV) were evaluated to determine their accuracy after storage at various temperatures/humidities. Rates of positive tests of control groups, experimental groups, and industry standards were compared (Fisher's exact chi2, p < or = 0.05). Compared to the control group, 2 of 10 HIV detection devices were adversely affected by exposure to high temperature/high humidity or high temperature/low humidity. With one exception, none of the environmentally exposed HCV or HBV detection devices exhibited significant differences compared to those stored under control conditions. For HIV, HCV, and HBV devices, there were differences compared to the industry standard. Collectively, this evaluation of pathogen detection kits revealed that diagnostic performance varies among products and storage conditions, and that the tested products cannot be considered to be approved for use to screen blood, plasma, cell, or tissue donors.

Removal of either N-glycan site from the envelope receptor binding domain of Moloney and Friend but not AKV mouse ecotropic gammaretroviruses alters receptor usage

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

Knoper, Ryan C.; Ferrarone, John; Yan Yuhe

2009-09-01

Three N-linked glycosylation sites were removed from the envelope glycoproteins of Friend, Moloney, and AKV mouse ecotropic gammaretroviruses: gs1 and gs2, in the receptor binding domain; and gs8, in a region implicated in post-binding cell fusion. Mutants were tested for their ability to infect rodent cells expressing 4 CAT-1 receptor variants. Three mutants (Mo-gs1, Mo-gs2, and Fr-gs1) infect NIH 3T3 and rat XC cells, but are severely restricted in Mus dunni cells and Lec8, a Chinese hamster cell line susceptible to ecotropic virus. This restriction is reproduced in ferret cells expressing M. dunni dCAT-1, but not in cells expressing NIHmore » 3T3 mCAT-1. Virus binding assays, pseudotype assays, and the use of glycosylation inhibitors further suggest that restriction is primarily due to receptor polymorphism and, in M. dunni cells, to glycosylation of cellular proteins. Virus envelope glycan size or type does not affect infectivity. Thus, host range variation due to N-glycan deletion is receptor variant-specific, cell-specific, virus type-specific, and glycan site-specific.« less

When Fruits and Vegetables Are Optional, Elementary School Children Choose Processed over Whole Offerings

ERIC Educational Resources Information Center

Amin, Sarah A.; Yon, Bethany A.; Taylor, Jennifer C.; Johnson, Rachel K.

2014-01-01

Purpose/Objectives: Increasing children's fruit and vegetable (FV) consumption is an important goal for the National School Lunch Program (NSLP). In 2012 the NSLP began requiring students to select a FV. The objective of this study was to compare children's FV choices in two school cafeteria environments a year before these new USDA regulations…

Rural versus urban Texas WIC participants' fruit and vegetable consumption.

PubMed

Ettienne-Gittens, Reynolette; McKyer, E Lisako J; Odum, Mary; Diep, Cassandra S; Li, Yan; Girimaji, Asha; Murano, Peter S

2013-01-01

To explore fruit and vegetable (F&V) consumption among urban and rural Black Texas WIC participants. The study included 673 WIC mothers and their 351 children from the TEXFAN study. T-tests, Pearson chi-squared test, and regressions were performed. Among children, differences were found for fruit consumption but not vegetable. Urban children were offered and consumed a greater variety of F&V. Mothers' F&V intake was related to age, pregnancy status, and breastfeeding status. This study provides insight into a subsection of the WIC population. Further research should be done regarding increasing the amount and variety of F&V consumed by WIC women and children.

Cloning, bacterial expression and crystallization of Fv antibody fragments

NASA Astrophysics Data System (ADS)

E´, Jean-Luc; Boulot, Ginette; Chitarra, V´ronique; Riottot, Marie-Madeleine; Souchon, H´le`ne; Houdusse, Anne; Bentley, Graham A.; Narayana Bhat, T.; Spinelli, Silvia; Poljak, Roberto J.

1992-08-01

The variable Fv fragments of antibodies, cloned in recombinant plasmids, can be expressed in bacteria as functional proteins having immunochemical properties which are very similar or identical with those of the corresponding parts of the parent eukaryotic antibodies. They offer new possibilities for the study of antibody-antigen interactions since the crystals of Fv fragments and of their complexes with antigen reported here diffract X-rays to a higher resolution that those obtained with the cognate Fab fragments. The Fv approach should facilitate the structural study of the combining site of antibodies and the further characterization of antigen-antibody interactions by site-directed mutagenesis experiments.

Construction and Validation of SRA-FV Need Assessment.

PubMed

Thornton, David; Knight, Raymond A

2015-08-01

This article describes the construction and testing of a newly designed instrument to assess psychological factors associated with increased rates of sexual recidivism. The new instrument (Structured Risk Assessment-Forensic Version or SRA-FV) was based on previous research using the SRA framework. This article describes the results of testing SRA-FV with a large sample (N = 566) of sexual offenders being evaluated for an early civil commitment program. SRA-FV was found to significantly predict sexual recidivism for both child molesters and rapists and to have incremental predictive value relative to two widely used static actuarial instruments (Static-99R; Risk Matrix 2000/S). © The Author(s) 2013.

Good Practices for Learning to Recognize Actions Using FV and VLAD.

PubMed

Wu, Jianxin; Zhang, Yu; Lin, Weiyao

2016-12-01

High dimensional representations such as Fisher vectors (FV) and vectors of locally aggregated descriptors (VLAD) have shown state-of-the-art accuracy for action recognition in videos. The high dimensionality, on the other hand, also causes computational difficulties when scaling up to large-scale video data. This paper makes three lines of contributions to learning to recognize actions using high dimensional representations. First, we reviewed several existing techniques that improve upon FV or VLAD in image classification, and performed extensive empirical evaluations to assess their applicability for action recognition. Our analyses of these empirical results show that normality and bimodality are essential to achieve high accuracy. Second, we proposed a new pooling strategy for VLAD and three simple, efficient, and effective transformations for both FV and VLAD. Both proposed methods have shown higher accuracy than the original FV/VLAD method in extensive evaluations. Third, we proposed and evaluated new feature selection and compression methods for the FV and VLAD representations. This strategy uses only 4% of the storage of the original representation, but achieves comparable or even higher accuracy. Based on these contributions, we recommend a set of good practices for action recognition in videos for practitioners in this field.

T-cell immunotherapy for human MK-1-expressing tumors using a fusion protein of the superantigen SEA and anti-MK-1 scFv antibody.

PubMed

Ueno, Aruto; Arakawa, Fumiko; Abe, Hironori; Matsumoto, Hisanobu; Kudo, Toshio; Asano, Ryutaro; Tsumoto, Kohei; Kumagai, Izumi; Kuroki, Motomu; Kuroki, Masahide

2002-01-01

The bacterial superantigen staphylococcal enterotoxin A (SEA) is an extremely potent activator of T lymphocytes when presented on major histocompatibility complex (MHC) class II molecules. To develop a tumor-specific superantigen for cancer therapy, we constructed a recombinant fusion protein of SEA and the single-chain variable fragment (scFv) of the FU-MK-1 antibody, which recognizes a glycoprotein antigen (termed MK-1 antigen) present on most carcinomas. We employed recombinant DNA techniques to fuse recombinant mutant SEA to an scFv antibody derived from FU-MK-1 and the resulting fusion protein (SEA/FUscFv) was produced by a bacterial expression system, purified with a metal-affinity column, and characterized for its MK-1-binding specificity and its antitumor activity. The SEA/FUscFv fusion protein retained the reactivity with MK-1-expressing tumor cells, introduced a specific cytotoxicity of lymphokine-activated killer T-cells to the tumor cells, and consequently suppressed the tumor growth in a SCID mouse xenograft model. This genetically engineered SEA/FUscFv fusion protein may serve as a potentially useful immunotherapeutic reagent for human MK-1-expressing tumors.

Factor Xa Activation of Factor V is of Paramount Importance in Initiating the Coagulation System: Lessons from a Tick Salivary Protein

PubMed Central

Schuijt, Tim J.; Bakhtiari, Kamran; Daffre, Sirlei; DePonte, Kathleen; Wielders, Simone J.H.; Marquart, J. Arnoud; Hovius, Joppe W.; van der Poll, Tom; Fikrig, Erol; Bunce, Matthew W.; Camire, Rodney M.; Nicolaes, Gerry A.F.; Meijers, Joost C.M.; van 't Veer, Cornelis

2013-01-01

Background Generation of active procoagulant cofactor FVa and its subsequent association with the enzyme FXa to form the prothrombinase complex is a pivotal initial event in blood coagulation and has been the subject of investigative effort, speculation and controversy. The current paradigm assumes that FV activation is initiated by limited proteolysis by traces of (meizo) thrombin. Methods and Results Recombinant tick salivary protein TIX-5 was produced and anticoagulant properties were studied using plasma, whole blood and purified systems. Here we report that TIX-5 specifically inhibits FXa-mediated FV activation involving the B-domain of FV and show that FXa activation of FV is pivotal for plasma and blood clotting. In line, tick feeding is impaired on TIX-5 immune rabbits displaying the in vivo importance of TIX-5. Conclusions Our data elucidate a unique molecular mechanism by which ticks inhibit the host's coagulation system. Based on our data we propose a revised blood coagulation scheme wherein direct FXa-mediated FV activation occurs in the initiation phase during which thrombin-mediated FV activation is restrained by fibrinogen and inhibitors. PMID:23817575

Structural Variation of Type I-F CRISPR RNA Guided DNA Surveillance.

PubMed

Pausch, Patrick; Müller-Esparza, Hanna; Gleditzsch, Daniel; Altegoer, Florian; Randau, Lennart; Bange, Gert

2017-08-17

CRISPR-Cas systems are prokaryotic immune systems against invading nucleic acids. Type I CRISPR-Cas systems employ highly diverse, multi-subunit surveillance Cascade complexes that facilitate duplex formation between crRNA and complementary target DNA for R-loop formation, retention, and DNA degradation by the subsequently recruited nuclease Cas3. Typically, the large subunit recognizes bona fide targets through the PAM (protospacer adjacent motif), and the small subunit guides the non-target DNA strand. Here, we present the Apo- and target-DNA-bound structures of the I-Fv (type I-F variant) Cascade lacking the small and large subunits. Large and small subunits are functionally replaced by the 5' terminal crRNA cap Cas5fv and the backbone protein Cas7fv, respectively. Cas5fv facilitates PAM recognition from the DNA major groove site, in contrast to all other described type I systems. Comparison of the type I-Fv Cascade with an anti-CRISPR protein-bound I-F Cascade reveals that the type I-Fv structure differs substantially at known anti-CRISPR protein target sites and might therefore be resistant to viral Cascade interception. Copyright © 2017 Elsevier Inc. All rights reserved.

Design and Evaluation of a Training Protocol for a Photographic Method of Visual Estimation of Fruit and Vegetable Intake among Kindergarten Through Second-Grade Students.

PubMed

Masis, Natalie; McCaffrey, Jennifer; Johnson, Susan L; Chapman-Novakofski, Karen

2017-04-01

To design a replicable training protocol for visual estimation of fruit and vegetable (FV) intake of kindergarten through second-grade students through digital photography of lunch trays that results in reliable data for FV served and consumed. Protocol development through literature and researcher input was followed by 3 laboratory-based trainings of 3 trainees. Lunchroom data collection sessions were done at 2 elementary schools for kindergarten through second-graders. Intraclass correlation coefficients (ICCs) were used. By training 3, ICC was substantial for amount of FV served and consumed (0.86 and 0.95, respectively; P < .05). The ICC was moderate for percentage of fruits consumed (0.67; P = .06). In-school estimates for ICCs were all significant for amounts served at school 1 and percentage of FV consumed at both schools. The protocol resulted in reliable estimation of combined FV served and consumed using digital photography. The ability to estimate FV intake accurately will benefit intervention development and evaluation. Copyright © 2017 Society for Nutrition Education and Behavior. Published by Elsevier Inc. All rights reserved.

Expression of single-chain Fv gene specific for gamma-seminoprotein by RTS and its biological activity identification.

PubMed

Han, Yuedong; Haun, Yi; Deng, Jinlan; Gao, Feng; Pan, Bifeng; Cui, Daxiang

2006-01-01

Fabricating a single-chain variable fragment specific for human seminoprotein is very important in antibody-directed enzyme prodrug therapy and NMR imaging for prostate cancer. Here a single-chain Fv specific for gamma-seminoprotein was expressed by RTS. Its activity and the efficiency of entry into prostate cancer cells are investigated by immunoprecipitation and Western blotting and immunofluorescent staining, as well as entry of conjugated magnetic beads into cells. Results showed that ScFv peptides specific for gamma-seminoprotein were successfully prepared, which can bind with the prostate cells specifically and can bring magnetic beads into prostate cancer cells within 15 min, the amount of magnetic beads inside prostate cancer cells increased as the culture time prolonged. ScFv-conjugated magnetic beads did not enter into control cells. In conclusion, the ScFv peptide against human gamma-seminoprotein with biological activity was successfully fabricated, which can take magnetic beads to prostate cancer cells specifically and not to the control cells. This ScFv peptide against human gamma-seminoprotein should be useful in improving the detection and therapy of prostate cancer at early stages and NMR imaging.

Applying normalization process theory to understand implementation of a family violence screening and care model in maternal and child health nursing practice: a mixed method process evaluation of a randomised controlled trial.

PubMed

Hooker, Leesa; Small, Rhonda; Humphreys, Cathy; Hegarty, Kelsey; Taft, Angela

2015-03-28

In Victoria, Australia, Maternal and Child Health (MCH) services deliver primary health care to families with children 0-6 years, focusing on health promotion, parenting support and early intervention. Family violence (FV) has been identified as a major public health concern, with increased prevalence in the child-bearing years. Victorian Government policy recommends routine FV screening of all women attending MCH services. Using Normalization Process Theory (NPT), we aimed to understand the barriers and facilitators of implementing an enhanced screening model into MCH nurse clinical practice. NPT informed the process evaluation of a pragmatic, cluster randomised controlled trial in eight MCH nurse teams in metropolitan Melbourne, Victoria, Australia. Using mixed methods (surveys and interviews), we explored the views of MCH nurses, MCH nurse team leaders, FV liaison workers and FV managers on implementation of the model. Quantitative data were analysed by comparing proportionate group differences and change within trial arm over time between interim and impact nurse surveys. Qualitative data were inductively coded, thematically analysed and mapped to NPT constructs (coherence, cognitive participation, collective action and reflexive monitoring) to enhance our understanding of the outcome evaluation. MCH nurse participation rates for interim and impact surveys were 79% (127/160) and 71% (114/160), respectively. Twenty-three key stakeholder interviews were completed. FV screening work was meaningful and valued by participants; however, the implementation coincided with a significant (government directed) change in clinical practice which impacted on full engagement with the model (coherence and cognitive participation). The use of MCH nurse-designed FV screening/management tools in focussed women's health consultations and links with FV services enhanced the participants' work (collective action). Monitoring of FV work (reflexive monitoring) was limited. The use of theory-based process evaluation helped identify both what inhibited and enhanced intervention effectiveness. Successful implementation of an enhanced FV screening model for MCH nurses occurred in the context of focussed women's health consultations, with the use of a maternal health and wellbeing checklist and greater collaboration with FV services. Improving links with these services and the ongoing appraisal of nurse work would overcome the barriers identified in this study.

Effects of garlic and diallyl trisulfide on the growth, photosynthesis, and alkaline phosphatase activity of the toxic cyanobacterium Microcystis aeruginosa.

PubMed

Wang, Shoubing; Wang, Yuanan; Ma, Xiaoxue; Xu, Ziran

2016-03-01

To identify a botanical algicide and elucidate the response of cyanobacteria to the extract from higher plants, the effects of garlic and garlic-derived diallyl trisulfide on Microcystis aeruginosa were studied. Effects were evaluated by changes in cell density, chlorophyll a, maximum effective quantum yield (Fv/Fm), effective quantum yield (YII), non-photochemical quenching (NPQ), and rapid light curves of M. aeruginosa. In addition, alkaline phosphatase activity (APA) was measured when M. aeruginosa was incubated with diallyl trisulfide. Results indicated that the inhibition by garlic and diallyl trisulfide was significant. The 120-h 50 % effective concentrations of garlic and diallyl trisulfide (EC50) were 0.75 g L(-1) and 2.84 mg L(-1), respectively. Moreover, the inhibitory rate increased with increasing concentration and the growth of M. aeruginosa was inhibited by 90.0 % at the highest concentrations. We also show that the response of M. aeruginosa to stress could involve both impairment of the photosynthetic center PSII and alteration of APA. For example, at high garlic concentration (2.0 g L(-1)), Fv/Fm significantly decreased from 0.501 to 0.084 (p < 0.05) after 120 h of exposure. Furthermore, the total APA was significantly decreased by exposure to a high diallyl trisulfide concentration after 24 h exposure. As new algal inhibitors, there are several advantages for their utilization, such as being common, cheap, non-toxic, and with high efficiency. It would be meaningful to further research on garlic as an environmentally friendly algicide.

Genome-wide identification and expression profile analysis of the NAC transcription factor family during abiotic and biotic stress in woodland strawberry

PubMed Central

Qi, Yanxiang; Liu, Xiaomei; Pu, Jinji

2018-01-01

The NAC transcription factors involved plant development and response to various stress stimuli. However, little information is available concerning the NAC family in the woodland strawberry. Herein, 37 NAC genes were identified from the woodland strawberry genome and were classified into 13 groups based on phylogenetic analysis. And further analyses of gene structure and conserved motifs showed closer relationship of them in every subgroup. Quantitative real-time PCR evaluation different tissues revealed distinct spatial expression profiles of the FvNAC genes. The comprehensive expression of FvNAC genes revealed under abiotic stress (cold, heat, drought, salt), signal molecule treatments (H2O2, ABA, melatonin, rapamycin), biotic stress (Colletotrichum gloeosporioides and Ralstonia solanacearum). Expression profiles derived from quantitative real-time PCR suggested that 5 FvNAC genes responded dramatically to the various abiotic and biotic stresses, indicating their contribution to abiotic and biotic stresses resistance in woodland strawberry. Interestingly, FvNAC genes showed greater extent responded to the cold treatment than other abiotic stress, and H2O2 exhibited a greater response than ABA, melatonin, and rapamycin. For biotic stresses, 3 FvNAC genes were up-regulated during infection with C. gloeosporioides, while 6 FvNAC genes were down-regulated during infection with R. solanacearum. In conclusion, this study identified candidate FvNAC genes to be used for the genetic improvement of abiotic and biotic stress tolerance in woodland strawberry. PMID:29897926

Healthy Foods, Healthy Families: combining incentives and exposure interventions at urban farmers' markets to improve nutrition among recipients of US federal food assistance.

PubMed

Bowling, April B; Moretti, Mikayla; Ringelheim, Kayla; Tran, Alvin; Davison, Kirsten

2016-01-01

Healthy Foods, Healthy Families (HFHF) is a fruit and vegetable (F&V) exposure/incentive program implemented at farmers' markets in low-income neighborhoods, targeting families receiving US federal food assistance. We examined program effects on participants' diet and associations between attendance, demographics and dietary change. Exposure activities included F&V tastings and cooking demonstrations. Incentives included 40% F&V bonus for electronic benefit transfer (EBT) card users and $20 for use purchasing F&V at every third market visit. Self-report surveys measuring nutritional behaviors/literacy were administered to participants upon enrollment (n = 425, 46.2% Hispanic, 94.8%female). Participants were sampled for follow-up at markets during mid-season (n = 186) and at season end (n = 146). Attendance was tracked over 16 weeks. Participants post-intervention reported significantly higher vegetable consumption(P = 0.005) and lower soda consumption (P = 0.005). Participants reporting largest F&V increases attended the market 6-8 times and received $40 in incentives. No change in food assistance spent on F&V (P = 0.94); 70% reported significant increases in family consumption of F&V,indicating subsidies increased overall F&V purchasing. Participants reported exposure activities and incentives similarly affected program attendance. Interventions combining exposure activities and modest financial incentives at farmers' markets in low-income neighborhoods show strong potential to improve diet quality of families receiving federal food assistance.

Development of a Novel Human Single Chain Antibody Against EGFRVIII Antigen by Phage Display Technology.

PubMed

Rahbarnia, Leila; Farajnia, Safar; Babaei, Hossein; Majidi, Jafar; Akbari, Bahman; Ahdi Khosroshahi, Shiva

2016-12-01

Purpose: EGFRvIII as the most common mutant variant of the epidermal growth factor receptor is resulting from deletion of exons 2-7 in the coding sequence and junction of exons 1 and 8 through a novel glycine residue. EGFRvIII is highly expressed in glioblastoma, carcinoma of the breast, ovary, and lung but not in normal cells. The aim of the present study was identification of a novel single chain antibody against EGFRvIII as a promising target for cancer therapy. Methods: In this study, a synthetic peptide corresponding to EGFRvIII protein was used for screening a naive human scFv phage library. A novel five-round selection strategy was used for enrichment of rare specific clones. Results: After five rounds of screening, six positive scFv clones against EGFRvIII were selected using monoclonal phage ELISA, among them, only three clones had expected size in PCR reaction. The specific interaction of two of the scFv clones with EGFRvIII was confirmed by indirect ELISA. One phage clone with higher affinity in scFv ELISA was purified for further analysis. The purity of the produced scFv antibody was confirmed using SDS-PAGE and Western blotting analyses. Conclusion: In the present study, a human anti- EGFRvIII scFv with high affinity was first identified from a scFv phage library. This study can be the groundwork for developing more effective diagnostic and therapeutic agents against EGFRvIII expressing cancers.

Salad Bars Increased Selection and Decreased Consumption of Fruits and Vegetables 1 Month After Installation in Title I Elementary Schools: A Plate Waste Study.

PubMed

Bean, Melanie K; Brady Spalding, Bethany; Theriault, Elizabeth; Dransfield, Kayla-Brooke; Sova, Alexandra; Dunne Stewart, Mary

2018-06-01

To evaluate the 1-month impact of salad bars on fruit and vegetable (FV) selection, intake, and waste. Pre-post quasi-experimental design. Title I elementary schools in a large, urban district in central Virginia. Students (grades 1-5; >95% African American) from 2 elementary schools participated in plate waste assessments (282 plates were rated at baseline, 443 at post-assessment); fourth- and fifth-grade students from 15 (of 18 eligible) schools (n = 1,193) responded to surveys. Digital imagery plate waste assessments were conducted before salad bars were installed (baseline) and 1 month afterward (post). Post-surveys examined student perceptions of salad bars. Fruit and vegetable selection, consumption, and waste. General linear models (without considering clustering) examined changes in outcomes, controlling for school. Frequencies and qualitative analyses were applied to survey data. At post, students selected more types of FVs (1.81-2.58; P < .001), although FV consumption decreased by 0.65 cups (P < .001). Given the smaller portions selected, there was less FV waste (0.27 cups; P < .001) at post. Students liked the ability to choose FV from salad bars. Short-term exposure to salad bars increased the number of FV students chose but decreased FV consumption. Additional strategies are needed to increase FV consumption. Copyright © 2018 Society for Nutrition Education and Behavior. Published by Elsevier Inc. All rights reserved.

Functional Consequences of Complementarity-determining Region Deactivation in a Multifunctional Anti-nucleic Acid Antibody*

PubMed Central

Lee, Jiyeon; Kim, Hye-Jin; Roh, Jooho; Seo, Youngsil; Kim, Minjae; Jun, Hye-Ryeong; Pham, Chuong D.; Kwon, Myung-Hee

2013-01-01

Many murine monoclonal anti-DNA antibodies (Abs) derived from mice models for systemic lupus erythematosus have additional cell-penetration and/or nucleic acid-hydrolysis properties. Here, we examined the influence of deactivating each complementarity-determining region (CDR) within a multifunctional anti-nucleic acid antibody (Ab) that possesses these activities, the catalytic 3D8 single chain variable fragment (scFv). CDR-deactivated 3D8 scFv variants were generated by replacing all of the amino acids within each CDR with Gly/Ser residues. The structure of 3D8 scFv accommodated single complete CDR deactivations. Different functional activities of 3D8 scFv were affected differently depending on which CDR was deactivated. The only exception was CDR1, located within the light chain (LCDR1); deactivation of LCDR1 abolished all of the functional activities of 3D8 scFv. A hybrid Ab, HW6/3D8L1, in which the LCDR1 from an unrelated Ab (HW6) was replaced with the LCDR1 from 3D8, acquired all activities associated with the 3D8 scFv. These results suggest that the activity of a multifunctional 3D8 scFv Ab can be modulated by single complete CDR deactivation and that the LCDR1 plays a crucial role in maintaining Ab properties. This study presents a new approach for determining the role of individual CDRs in multifunctional Abs with important implications for the future of Ab engineering. PMID:24155236

5-a-day fruit and vegetable food product labels: reduced fruit and vegetable consumption following an exaggerated compared to a modest label.

PubMed

Appleton, K M; Pidgeon, H J

2018-05-15

Food product labels based on the WHO 5-a-day fruit and vegetable (FV) message are becoming increasingly common, but these labels may impact negatively on complementary or subsequent FV consumption. This study aimed to investigate the impact of a '3 of your 5-a-day' versus a '1 of your 5-a-day' smoothie product label on subsequent FV consumption. Using an acute experimental design, 194 participants (90 males, 104 females) were randomised to consume a smoothie labelled as either '3 of your 5-a-day' (N = 97) or '1 of your 5-a-day' (N = 97) in full, following a usual breakfast. Subsequent FV consumption was measured for the rest of the day using 24-h recall. Usual FV consumption was also assessed via 24-h recall for the day before the study. Regression analyses revealed a significantly lower subsequent FV consumption following smoothies displaying the '3 of your 5-a-day' label compared to the '1 of your 5-a-day' label (Beta = - 0.15, p = 0.04). Secondary analyses revealed these effects to be driven mainly by changes to consumption in usual high FV consumers, in females and in vegetable as opposed to fruit consumption. These findings demonstrate a role for label information in food intake, and the potential negative impacts of an exaggerated food product label on healthy food consumption and healthy dietary profiles.

An anti-CD30 single-chain Fv selected by phage display and fused to Pseudomonas exotoxin A (Ki-4(scFv)-ETÁ) is a potent immunotoxin against a Hodgkin-derived cell line

PubMed Central

Klimka, A; Barth, S; Matthey, B; Roovers, R C; Lemke, H; Hansen, H; Arends, J-W; Diehl, V; Hoogenboom, H R; Engert, A

1999-01-01

The human CD30 receptor is highly overexpressed on the surface of Hodgkin Reed-Sternberg cells and has been shown to be an excellent target for selective immunotherapy using monoclonal antibody-based agents such as immunotoxins. To construct a new recombinant immunotoxin for possible clinical use in patients with Hodgkin's lymphoma, we have chosen the murine anti-CD30 hybridoma Ki-4 to generate a high-affinity Ki-4 single-chain variable fragment (scFv). Hybridoma V-genes were polymerase chain reaction-amplified, assembled, cloned and expressed as a mini-library for display on filamentous phage. Functional Ki-4 scFv were obtained by selection of binding phage on the Hodgkin lymphoma-derived, CD30-expressing cell line L540Cy. The selected recombinant Ki-4 scFv was shown to specifically bind to an overlapping epitope on the CD30 antigen with binding kinetics similar to those of the original antibody. The Ki-4 scFv was subsequently fused to a deletion mutant of Pseudomonas exotoxin A (ETÁ). The resulting immunotoxin Ki-4(scFv)-ETÁ specifically binds to CD30+ L540Cy cells and inhibits the protein synthesis by 50% at a concentration (IC50) of 43 pM. This recombinant immunotoxin is a promising candidate for further clinical evaluation in patients with Hodgkin's lymphoma or other CD30+ malignancies. © 1999 Cancer Research Campaign PMID:10376974

Fluoride Varnish for Caries Prevention: Efficacy and Implementation.

PubMed

Bonetti, Debbie; Clarkson, Jan E

2016-01-01

Many reviews support fluoride varnish (FV) as a caries-inhibitory agent. Evidence from 6 Cochrane systematic reviews involving 200 trials and more than 80,000 participants further confirms the effectiveness of FV, applied professionally 2-4 times a year, for preventing dental caries in both primary and permanent teeth. The relative benefit of FV application seems to occur irrespective of baseline caries risk, baseline caries severity, background exposure to fluorides, use of fluoride toothpaste and application features such as prior prophylaxis, concentration of fluoride or frequency of application. While the efficacy of FV is acknowledged in clinical practice guidelines globally, the implementation of this recommendation may still be an issue. Factors that may facilitate FV application in the USA include Medicaid eligibility, relationships with dentists/community centers and strong cooperation and communication between physicians and support staff. Barriers include insufficient time to integrate oral health services into well-child visits, difficulty in applying FV (lack of skills/training) and resistance among colleagues and staff. Research in the UK/Scotland also suggests encouraging clinicians in their motivation to perform this treatment and addressing professional and parental concerns relating to possible negative consequences may be influential. Further research targeting cost-effectiveness and how FV in routine care may fit in with political agendas relating to, for example, inequalities in health care provision and access will also play a key part in stakeholder decisions to put resources into this issue. © 2016 The Author(s) Published by S. Karger AG, Basel.

Isolation and characterization of anti c-met single chain fragment variable (scFv) antibodies.

PubMed

Qamsari, Elmira Safaie; Sharifzadeh, Zahra; Bagheri, Salman; Riazi-Rad, Farhad; Younesi, Vahid; Abolhassani, Mohsen; Ghaderi, Sepideh Safaei; Baradaran, Behzad; Somi, Mohammad Hossein; Yousefi, Mehdi

2017-12-01

The receptor tyrosine kinase (RTK) Met is the cell surface receptor for hepatocyte growth factor (HGF) involved in invasive growth programs during embryogenesis and tumorgenesis. There is compelling evidence suggesting important roles for c-Met in colorectal cancer proliferation, migration, invasion, angiogenesis, and survival. Hence, a molecular inhibitor of an extracellular domain of c-Met receptor that blocks c-Met-cell surface interactions could be of great thera-peutic importance. In an attempt to develop molecular inhibitors of c-Met, single chain variable fragment (scFv) phage display libraries Tomlinson I + J against a specific synthetic oligopeptide from the extracellular domain of c-Met receptor were screened; selected scFv were then characterized using various immune techniques. Three c-Met specific scFv (ES1, ES2, and ES3) were selected following five rounds of panning procedures. The scFv showed specific binding to c-Met receptor, and significantly inhibited proliferation responses of a human colorectal carcinoma cell line (HCT-116). Moreover, anti- apoptotic effects of selected scFv antibodies on the HCT-116 cell line were also evaluated using Annexin V/PI assays. The results demonstrated rates of apoptotic cell death of 46.0, 25.5, and 37.8% among these cells were induced by use of ES1, ES2, and ES3, respectively. The results demonstrated ability to successfully isolate/char-acterize specific c-Met scFv that could ultimately have a great therapeutic potential in immuno-therapies against (colorectal) cancers.

Construction and characterization of a highly reactive chicken-derived single-chain variable fragment (scFv) antibody against Staphylococcus aureus developed with the T7 phage display system.

PubMed

Li, Jingquan; Xu, Yongping; Wang, Xitao; Li, Yuan; Wang, Lili; Li, Xiaoyu

2016-06-01

The purpose of this study was to construct a single-chain variable fragment (scFv) antibody from chicken egg yolk immunoglobulin (IgY) by means of genetic engineering and subsequent panning for a specific antibody against Staphylococcus aureus. We amplified the scFv using blood and spleen obtained from 100-day-old Roman chickens immunized with inactivated S. aureus and subsequently constructed a T7 phage display antibody library using phage display technology. Four non-repeated blood scFv and 6 spleen scFv were obtained following 3 rounds of panning of the T7 phage display antibody library, enzyme-linked immunosorbent assay and sequencing. These 10 scFv were cloned into the prokaryotic expression vector pCold I with expression induced at a low temperature. Four soluble proteins were obtained. Among them, soluble protein SFV6 derived from the spleen showed good reactivity against S. aureus using indirect ELISA and produced a particularly strong antibacterial effect in vitro. We were successful in isolating a highly specific scFv antibody against S. aureus from the spleen phage display library. This study provides a simple and rapid method for the quick preparation of a large number of antibodies against S. aureus and provides the foundation for the positioning of antibodies in the organism and the study of the antibacterial mechanism through which the antibody functions. Copyright © 2016 Elsevier B.V. All rights reserved.

Escherichia coli surface display of single-chain antibody VRC01 against HIV-1 infection

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

Wang, Lin-Xu; School of Biological Sciences, University of Nebraska—Lincoln, Lincoln, NE 68583; Mellon, Michael

Human immunodeficiency virus type 1 (HIV-1) transmission and infection occur mainly via the mucosal surfaces. The commensal bacteria residing in these surfaces can potentially be employed as a vehicle for delivering inhibitors to prevent HIV-1 infection. In this study, we have employed a bacteria-based strategy to display a broadly neutralizing antibody VRC01, which could potentially be used to prevent HIV-1 infection. The VRC01 antibody mimics CD4-binding to gp120 and has broadly neutralization activities against HIV-1. We have designed a construct that can express the fusion peptide of the scFv-VRC01 antibody together with the autotransporter β-barrel domain of IgAP gene frommore » Neisseria gonorrhoeae, which enabled surface display of the antibody molecule. Our results indicate that the scFv-VRC01 antibody molecule was displayed on the surface of the bacteria as demonstrated by flow cytometry and immunofluorescence microscopy. The engineered bacteria can capture HIV-1 particles via surface-binding and inhibit HIV-1 infection in cell culture. - Highlights: • Designed single-chain VRC01 antibody was demonstrated to bind HIV-1 envelope gp120. • Single-chain VRC01 antibody was successfully displayed on the surface of E. coli. • Engineered bacteria can absorb HIV-1 particles and prevent HIV-1 infection in cell culture.« less

Detection of E. coli O157:H7 with a reporter phage containing the luxCDABE cassette

USDA-ARS?s Scientific Manuscript database

Bacteriophage and reporter phage are used for typing and/or detection of pathogens. The temperate tailed phage fV10 has been utilized for phage-typing E. coli O157:H7. By modifying fV10 to transduce kanamycin resistance and the a luxCDABE cassette, we developed a reporter bacteriophage (fV10-lux) p...

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

Generation and evaluation of antibody agents for molecular imaging of CD44v6-expressing cancers

PubMed Central

Haylock, Anna-Karin; Nilvebrant, Johan; Mortensen, Anja; Velikyan, Irina; Nestor, Marika; Falk, Ronny

2017-01-01

Aim The aim of this study was to generate and characterize scFv antibodies directed to human CD44v6, as well as to radiolabel and evaluate top candidates in vitro and in vivo for their potential use in CD44v6-targeted molecular imaging in cancer patients. Materials and methods Phage display selections were used to isolate CD44v6-specific scFvs. A chain shuffling strategy was employed for affinity maturation based on a set of CD44v6-specific first-generation clones. Two second-generation scFv clones were then chosen for labeling with 111In or 125I and assessed for CD44v6-specific binding on cultured tumor cells. In vivo uptake and distribution was evaluated in tumor-bearing mice using a dual tumor model. Finally, a proof-of-concept small animal PET-CT study was performed on one of the candidates labeled with 124I. Results Two affinity-matured clones, CD44v6-scFv-A11 and CD44v6-scFv-H12, displayed promising binding kinetics. Seven out of eight radiolabeled conjugates demonstrated CD44v6-specific binding. In vivo studies on selected candidates demonstrated very advantageous tumor-to-organ ratios, in particular for iodinated conjugates, where 125I-labeled scFvs exhibited favorable kinetics and tumor-to-blood ratios above five already at 24 hours p.i.. The small animal PET-CT study using 124I-labeled CD44v6-scFv-H12 was in line with the biodistribution data, clearly visualizing the high CD44v6-expressing tumor. Conclusion The single chain fragments, CD44v6-scFv-A11 and CD44v6-scFv-H12 specifically bind to CD44v6, and the radiolabeled counterparts provide high tumor-to-blood ratios and fast clearance from organs and blood. We conclude that radioiodinated CD44v6-scFv-A11 and CD44v6-scFv-H12 possess features highly suitable for stringent molecular imaging. PMID:29029420

Relevance of fruits, vegetables and flavonoids from fruits and vegetables during early life, mid-childhood and adolescence for levels of insulin-like growth factor (IGF-1) and its binding proteins IGFBP-2 and IGFBP-3 in young adulthood.

PubMed

Krupp, Danika; Remer, Thomas; Penczynski, Katharina J; Bolzenius, Katja; Wudy, Stefan A; Buyken, Anette E

2016-02-14

The growth hormone (GH) insulin-like growth factor (IGF) axis has been linked to insulin metabolism and cancer risk. Experimental evidence indicates that the GH-IGF axis itself can be influenced by dietary flavonoids. As fruit and vegetable (FV) intake is a major source of flavonoid consumption, FV's beneficial health effects may be explained via flavonoids' influence on the GH-IGF axis, but observational evidence is currently rare. We used data from Dortmund Nutritional and Anthropometric Longitudinally Designed Study participants to analyse prospective associations between FV, fruit intake and flavonoid intake from FV (FlavFV) with IGF-1 and its binding proteins IGFBP-2 and IGFBP-3. Subjects needed to provide a fasting blood sample in adulthood (18-39 years) and at least two 3-d weighed dietary records in early life (0·5-2 years, n 191), mid-childhood (3-7 years, n 265) or adolescence (girls: 9-15 years, boys: 10-16 years, n 261). Additional analyses were conducted among those providing at least three 24-h urine samples in adolescence (n 236) to address the predictor urinary hippuric acid (HA), a biomarker of polyphenol intake. Higher fruit intake in mid-childhood and adolescence was related to higher IGFBP-2 in adulthood (P=0·03 and P=0·045). Comparable trends (P=0·045-0·09) were discernable for FV intake (but not FlavFV) in all three time windows. Similarly, higher adolescent HA excretion tended to be related (P=0·06) to higher adult IGFBP-2 levels. Regarding IGFBP-3, a marginal (P=0·08) positive association was observed with FlavFV in mid-childhood only. None of the investigated dietary factors was related to IGF-1. In conclusion, higher fruit and FV intakes during growth may be relevant for adult IGFBP-2, but probably not for IGFBP-3 or IGF-1.

Fresh Fruit and Vegetable Program and Requests for Fruits and Vegetables Outside School Settings.

PubMed

Ohri-Vachaspati, Punam; Dachenhaus, Elizabeth; Gruner, Jessie; Mollner, Kristina; Hekler, Eric B; Todd, Michael

2018-01-08

Consumption of fruits and vegetables (F/V) among elementary school-aged children remains inadequate, especially among low-income children. The US Department of Agriculture's Fresh Fruit and Vegetable Program (FFVP) provides F/V as snacks to children during the school day, outside of school meals. School-based initiatives are successful in changing behaviors in school settings; however, their influence on behaviors outside of schools needs investigation. To examine whether FFVP participation is associated with F/V requests at stores, self-efficacy to ask for and choose F/V at home, and F/V consumption. Cross-sectional study. Fourth graders in six classrooms (n=296) from three urban, low-income school districts in Phoenix, AZ, were surveyed during 2015; one FFVP and one non-FFVP school from each district that were similar in school size, percent free/reduced-price meal eligibility, and race/ethnicity of enrolled students were selected. Children's self-reported F/V requests during shopping, their self-efficacy to ask for and choose F/V at home, and F/V consumption on the previous day (non-FFVP school day) were measured using questions adapted from validated surveys. Multivariable mixed-effect regression models, adjusting for clustering of students within classes and classes within schools were explored. In models adjusting for individual-level factors (ie, age and sex) only, several significant positive associations were observed between school FFVP participation and healthier F/V outcomes. After additionally adjusting for school-level factors (ie, total enrollment and % Hispanic/Latino students) significant associations were observed between school FFVP participation and more requests for vegetables during shopping (P<0.001), higher scores on self-efficacy to choose vegetables at home (P=0.004), stronger preferences for vegetables (P<0.001), and more frequent consumption of fruit (P=0.006). School FFVP participation was associated with more requests for vegetables during shopping and higher self-efficacy to make healthy choices at home, suggesting the influence of the FFVP may extend beyond the school day. Copyright © 2018 Academy of Nutrition and Dietetics. Published by Elsevier Inc. All rights reserved.

Feasibility and Acceptability of Brighter Bites: A Food Co-Op in Schools to Increase Access, Continuity and Education of Fruits and Vegetables Among Low-Income Populations.

PubMed

Sharma, Shreela; Helfman, Lisa; Albus, Katherine; Pomeroy, Mike; Chuang, Ru-Jye; Markham, Christine

2015-08-01

Intake of fruits and vegetables (F&V) continues to be low in children in the United States. The purpose of this study was to conduct a pilot feasibility evaluation of Brighter Bites, a school-based food co-op to provide access to fresh F&V and nutrition education to low-income children and their families. Brighter Bites is a 16-week school-based food co-op consisting of: (1) Weekly distribution of 50-60 servings of fresh F&V; (2) Weekly bilingual parent handouts and recipe demonstrations; and (3) implementing CATCH, a coordinated school health program in schools. Brighter Bites was pilot tested using a pre-post evaluation design in one charter school in Houston, TX, USA (n = 57 3rd grade parent-child dyads; 94.1 % Hispanic, 91 % low-income). Evaluation, at baseline, midpoint, and post-intervention, included self-reported child and parent surveys on psychosocial factors, dietary habits and mealtime practices. Pearson's Chi square test, Fisher's exact-test or paired t test were used to determine changes pre- to post-intervention (at p < 0.05). Process data using parent surveys, teacher surveys, attendance logs, and produce cost data were used to determine feasibility and acceptability of program. Participants received on average 61 servings of F&V weekly for 16 weeks at the cost of $4.31/family/week. Results showed significant increases in child reported self-efficacy, outcome expectations and attitudes towards consuming F&V (p < 0.05). We found significant increases in child exposure to F&V and child preference of various F&V from baseline to post-intervention (p < 0.05). Parent surveys showed significant improvements in mealtime practices at home: decrease in children eating while watching TV, increase in eating dinner with the family, less fast food, less sugary drinks with meals, more children asking for F&V as snacks. Process data showed 98 % retention rate and high parent acceptability of program components. Brighter Bites is a promising strategy to increase F&V access and education in low-income populations using existing infrastructure of schools and food banks.

Free stream capturing in fluid conservation law for moving coordinates in three dimensions

NASA Technical Reports Server (NTRS)

Obayashi, Shigeru

1991-01-01

The free-stream capturing technique for both the finite-volume (FV) and finite-difference (FD) framework is summarized. For an arbitrary motion of the grid, the FV analysis shows that volumes swept by all six surfaces of the cell have to be computed correctly. This means that the free-stream capturing time-metric terms should be calculated not only from a surface vector of a cell at a single time level, but also from a volume swept by the cell surface in space and time. The FV free-stream capturing formulation is applicable to the FD formulation by proper translation from an FV cell to an FD mesh.

Affinity improvement of a therapeutic antibody to methamphetamine and amphetamine through structure-based antibody engineering

PubMed Central

Thakkar, Shraddha; Nanaware-Kharade, Nisha; Celikel, Reha; Peterson, Eric C.; Varughese, Kottayil I.

2014-01-01

Methamphetamine (METH) abuse is a worldwide threat, without any FDA approved medications. Anti-METH IgGs and single chain fragments (scFvs) have shown efficacy in preclinical studies. Here we report affinity enhancement of an anti-METH scFv for METH and its active metabolite amphetamine (AMP), through the introduction of point mutations, rationally designed to optimize the shape and hydrophobicity of the antibody binding pocket. The binding affinity was measured using saturation binding technique. The mutant scFv-S93T showed 3.1 fold enhancement in affinity for METH and 26 fold for AMP. The scFv-I37M and scFv-Y34M mutants showed enhancement of 94, and 8 fold for AMP, respectively. Structural analysis of scFv-S93T:METH revealed that the substitution of Ser residue by Thr caused the expulsion of a water molecule from the cavity, creating a more hydrophobic environment for the binding that dramatically increases the affinities for METH and AMP. PMID:24419156

A Comparison of Smokers' and Nonsmokers' Fruit and Vegetable Intake and Relevant Psychosocial Factors

PubMed Central

McClure, Jennifer B.; Divine, George; Alexander, Gwen; Tolsma, Dennis; Rolnick, Sharon J.; Stopponi, Melanie; Richards, Julie; Johnson, Christine C.

2009-01-01

The authors examined the relation between smoking status and fruit and vegetable (FV) consumption among a population-based sample and examined differences in psychosocial factors that may influence diet and inform intervention efforts. The authors recruited adults (N = 2,540) from 5 US health plans to participate in a Web-based dietary intervention trial. At baseline, smokers ate fewer FV servings per day (p < .001) and were less likely to meet the 5 A Day goal (p < .001). Smokers reported lower self-efficacy, overall motivation, and intrinsic motivation for meeting daily FV recommendations. Fewer smokers expected that eating 5 FV servings a day would reduce their risk for diabetes (p = .02) or obesity (p = .008). Smokers are an important target group for dietary intervention. Intervention efforts should attempt to increase smokers' motivation and confidence in their abilities to change their eating patterns and educate them about the health benefits of eating FV. PMID:19297300

Fv-clasp: An Artificially Designed Small Antibody Fragment with Improved Production Compatibility, Stability, and Crystallizability.

PubMed

Arimori, Takao; Kitago, Yu; Umitsu, Masataka; Fujii, Yuki; Asaki, Ryoko; Tamura-Kawakami, Keiko; Takagi, Junichi

2017-10-03

Antibody fragments are frequently used as a "crystallization chaperone" to aid structural analysis of complex macromolecules that are otherwise crystallization resistant, but conventional fragment formats have not been designed for this particular application. By fusing an anti-parallel coiled-coil structure derived from the SARAH domain of human Mst1 kinase to the variable region of an antibody, we succeeded in creating a novel chimeric antibody fragment of ∼37 kDa, termed "Fv-clasp," which exhibits excellent crystallization compatibility while maintaining the binding ability of the original IgG molecule. The "clasp" and the engineered disulfide bond at the bottom of the Fv suppressed the internal mobility of the fragment and shielded hydrophobic residues, likely contributing to the high heat stability and the crystallizability of the Fv-clasp. Finally, Fv-clasp antibodies showed superior "chaperoning" activity over conventional Fab fragments, and facilitated the structure determination of an ectodomain fragment of integrin α6β1. Copyright © 2017 Elsevier Ltd. All rights reserved.

Bullying Victimization Among Chinese Middle School Students: The Role of Family Violence.

PubMed

Zhu, Yuhong; Chan, Ko Ling; Chen, Jinsong

2018-06-01

This study used the data from a representative sample to investigate the association between family violence (FV) and child bullying victimization (BV) in Xi'an city, China. Data on social demographic information and the prevalence of BV and FV were collected from a randomly selected sample with 3,175 middle school students aged 15 to 17 by self-administrated questionnaires. Results show that 55.9% and 30.3% of the participants have witnessed intimate partner violence (IPV), 37.7% and 30.8% have been victims of child abuse, and 54.9% and 44.6% have been bullied in a lifetime and in the preceding year, respectively. The lifetime and preceding-year co-occurrence rate of FV and BV are 45% and 30.4%, respectively. Multiple logistic regressions confirm FV as a unique risk factor in predicting both direct and relational BV after controlling for a number of confounding factors. This study suggests that FV experiences should be included in the screening and assessment of risk for child BV.

Who are eating and not eating fruits and vegetables in Malaysia?

PubMed

Yen, Steven T; Tan, Andrew K G

2012-12-01

To investigate the roles of sociodemographic factors in fruit and vegetable (FV) consumption in Malaysia. Data are obtained from the Malaysia Non-Communicable Disease Surveillance-1. Logistic regressions are conducted using a multiracial (Malay, Chinese, Indian and other ethnic groups) sample of 2,447 observations to examine the factors affecting individual decisions to consume FV on a daily basis. Based on the binary outcomes of whether individuals consumed FV daily, results indicate that work hours, education, age ethnicity, income, gender, smoking status, and location of residence are significantly correlated with daily fruit consumption. Daily vegetable consumption is significantly correlated with income, gender, health condition, and location of residence. Our results imply the need for programs to educate and motivate consumers to make healthier dietary choices. Interventions to increase FV consumption by changing behaviors should be considered, as should those that increase public awareness of the dietary benefits of FV. These intervention programs should be targeted at and tailored toward individuals who are less educated, younger, less affluent, males, smokers, and metropolitan dwellers.

Consumption of fruits and vegetables in Malaysia: profiling the daily and nondaily consumers.

PubMed

Yen, Steven T; Tan, Andrew K G; Feisul, Mustapha I

2015-03-01

This study examines the sociodemographic factors associated with daily fruit and vegetable (FV) consumption in Malaysia. Based on a cross-sectional sample of 2447 individuals from the Malaysia Non-Communicable Disease Surveillance-1, a multivariate sample selection system is developed and estimated, to accommodate high frequencies of daily FV consumption and the days of servings among nondaily consumers. Results indicate that the authors' attempt to account for endogenous sample selectivity and cross-equation correlations is justified. There exist positive correlations between FV consumption likelihoods and longer work hours, higher levels of education, high income, female gender, nonsmoking status, and being from East Malaysia. Among nondaily consumers, those with longer work hours, singles, and people with diabetes are less inclined to eat fruits on more days. Overall, higher-educated, affluent people, nonsmokers, and East Malaysians display more days of FV consumption. Based on these outcomes, several policy implications are recommended vis-à-vis FV consumption patterns in Malaysia. © 2012 APJPH.

Effects of parenting style and parent-related weight and diet on adolescent weight status.

PubMed

Alia, Kassandra A; Wilson, Dawn K; St George, Sara M; Schneider, Elizabeth; Kitzman-Ulrich, Heather

2013-04-01

This study examined the interaction between parental limit setting of sedentary behaviors and health factors (weight status, physical activity [PA], fruit and vegetable [FV] intake) on standardized body mass index (zBMI) in African American adolescents. Data were from 67 parent-adolescent dyads. Parental limit setting, PA and FV intake were assessed via self-report, and objective height and weight measurements were collected. Regressions examined the interaction between parental limit setting and BMI, PA, FV intake on adolescent zBMI. The model for parent BMI and FV intake accounted for 31% of the variance in adolescent zBMI. A significant interaction for parent BMI by limit setting showed that as parental BMI increased, higher (vs. lower) limit setting was associated with lower adolescent zBMI. Higher parent FV consumption was associated with lower adolescent zBMI. Future interventions should integrate parent limit setting and target parent fruit and vegetable intake for obesity prevention in underserved adolescents.

Harmful effects behind the daily supplementation of a fixed vegetarian blend in the rat model.

PubMed

Bonamassa, Barbara; Canistro, Donatella; Sapone, Andrea; Vivarelli, Fabio; Vornoli, Andrea; Longo, Vincenzo; Paolini, Moreno

2016-11-01

Fruit and vegetables (FV) have long been considered a panacea against major chronic diseases, including cancer. However, there is no convincing epidemiological, clinical or experimental evidence supporting FV chemopreventive ability. A daily mono-supplementation of lyophilized onion, tomato, peach, black grape or lettuce was compared with the daily combined administration of the same FV (5 a day-like diet). Ten days post-treatment, the phase-I/II xenobiotic metabolizing and antioxidant enzyme activities, protein and mRNA levels were investigated. As a marker of oxidative stress, the level of hydroperoxides was measured in rat serum samples. Here we show that a blend of FV orally administered to rats not only potentially manipulates metabolism but also disrupts systemic oxidative homeostasis. A daily combination of the five servings remarkably down-regulates the catalytic activity, protein and mRNA levels of a cohort of hepatic metabolizing enzymes, suggesting a possible depressed clearance upon exposure to ubiquitous carcinogens. Strikingly, we observed an impairment of antioxidant enzymes with a boost in systemic hydroperoxide levels. Our study identifies new potential factors of cancer risk connected with the persistent consumption of fixed servings of FV, suggesting that dietary guidance should rely on a "daily diversification" of FV. Copyright © 2016 Elsevier Ltd. All rights reserved.

Family Violence and Other Potentially Traumatic Interpersonal Events Among 9- to 17-Year-Old Children Attending an Outpatient Psychiatric Clinic.

PubMed

Hultmann, Ole; Broberg, Anders G

2016-11-01

Among children visiting child and adolescent mental health care (CAM), the prevalence of exposure to family violence (FV) is reported to exceed prevalence in community samples, as are potentially traumatic interpersonal events (IPE) outside the family. The aim of the study was to relate CAM patients' self-reported experiences of violence exposure to their current psychiatric symptoms and to compare patients exposed to violence with patients who reported no exposure. We asked 305 consecutive 9- to 17-year-old patients in CAM about their current and previous exposure to violence in and outside of the family. Prevalence of exposure to any kind of violence was 67%. Reported exposures were 19% to IPE, 21% to FV, and 27% to both. Children exposed to both FV and IPE were more negatively affected by the events than children exposed to FV or IPE only. Children in the FV + IPE group reported more mental health symptoms than those in the no violence (33%) group. In general, IPE was related to the outcome measures only in combination with FV. Degree of violence exposure seemed to have a dose-response relationship with the diagnosis of post-traumatic stress disorder. © The Author(s) 2015.

Analysis of the relationship between end-to-end distance and activity of single-chain antibody against colorectal carcinoma.

PubMed

Zhang, Jianhua; Liu, Shanhong; Shang, Zhigang; Shi, Li; Yun, Jun

2012-08-22

We investigated the relationship of End-to-end distance between VH and VL with different peptide linkers and the activity of single-chain antibodies by computer-aided simulation. First, we developed (G4S)n (where n = 1-9) as the linker to connect VH and VL, and estimated the 3D structure of single-chain Fv antibody (scFv) by homologous modeling. After molecular models were evaluated and optimized, the coordinate system of every protein was built and unified into one coordinate system, and End-to-end distances calculated using 3D space coordinates. After expression and purification of scFv-n with (G4S)n as n = 1, 3, 5, 7 or 9, the immunoreactivity of purified ND-1 scFv-n was determined by ELISA. A multi-factorial relationship model was employed to analyze the structural factors affecting scFv: rn=ABn-ABO2+CDn-CDO2+BCn-BCst2. The relationship between immunoreactivity and r-values revealed that fusion protein structure approached the desired state when the r-value = 3. The immunoreactivity declined as the r-value increased, but when the r-value exceeded a certain threshold, it stabilized. We used a linear relationship to analyze structural factors affecting scFv immunoreactivity.

LA Sprouts: A 12-week gardening, nutrition, and cooking randomized control trial improves determinants of dietary behaviors

PubMed Central

Davis, Jaimie N; Martinez, Lauren C.; Spruijt-Metz, Donna; Gatto, Nicole M.

2015-01-01

Objective To evaluate the effect of an exploratory 12-week nutrition, cooking and gardening RCT (“LA Sprouts”) on preference for fruit and vegetables (FV); willingness to try FV; identification of FV; self-efficacy to garden/eat/cook FV; motivation to garden/eat/cook FV; attitudes towards FV; nutrition and gardening knowledge; and home gardening habits. Design and Participants Four elementary schools with 304 predominately Hispanic/Latino 3rd–5th grade students were randomized to either the LA Sprouts (n=167 students) or Control group (n=137 students). LA Sprouts participants received 12 weeks of weekly 90-minute culturally tailored gardening, nutrition, and cooking classes after school. Questionnaire data examining dietary determinants were obtained at baseline and post-intervention. Results After the 12-week program, LA Sprouts participants compared with controls improved scores for identification of vegetables (+11% vs. +5%; P=.001), nutrition and gardening knowledge (+14.5% vs. −5.0%; P =.003), and were more likely to garden at home (+7.5% vs. −4.4%; P=.003). Conclusions The LA Sprouts program positively impacted a number of determinants of dietary behaviors, which suggest possible mechanisms by which gardening and nutrition education act to improve dietary intake and health outcomes. PMID:26453367

[Construction, expression and characterization of the fusion gene of super-antigen SEA and single chain Fv of the ND-1 monoclonal antibody against human colorectal cancer].

PubMed

Chen, Hang; Li, Li; Fang, Jin

2012-04-01

To construct and express the recombinant ND-1-scFv/SEA, a fusion protein of superantigen (staphylococcal enterotoxinA, SEA) and single-chain variable fragment of monoclonal antibody ND-1 against human clolorectal carcinoma, and to enhance the targeted killing effect of SEA. The expression of the fusion protein was induced in E.coli M15 by IPTG. Ni-NTA resin affinity chromatography was used to separate and purify the expressed product. The specific binding activity of the purified ND-1-scFv/SEA protein was examined by indirect immunofluorescence assay and the targeted-cytotoxicity was determined using MTT assay. The expressing vector of fusion gene ND-1scFv/SEA was constructed successfully. ND-1-scFv/SEA protein retained a high binding affinity to antigen-positive human colorectal cancer cell CCL-187 and had a stronger capability to activate PBMC and kill the target cells compared to SEA alone, with a killing rate of 91% at 4 μg/mL. ND-1-scFv/SEA fusion protein could specifically target colorectal cancer cell, enhance the activity of kill tumor cell and has potential applications in the targeted therapy of colorectal cancer.

"5 A Day" achievement badge for urban boy scouts: formative evaluation results.

PubMed

Cullen, K W; Baranowski, T; Baranowski, J; Warnecke, C; de Moor, C; Nwachokor, A; Hajek, R A; Jones, L A

1998-01-01

Certain cancers are more common among African Americans (AA). Fruit and vegetables (F&V) reduce cancer risk, but Americans, and African Americans in particular, do not meet the "5 A Day" goal. Scouting organizations, particularly urban Boy Scout groups that target inner-city youth, provide promising channels for nutritional behavioral change programs. Focus groups were conducted with urban Boy Scouts and their parents to identify factors influencing F&V consumption and evaluate potential intervention activities. Twenty-four-hour dietary recalls were collected from 85 area Boy Scouts. A national data set was used to obtain values for F&V consumption by African American and European American (boys age 0-16). Vegetable preferences were low and a negative peer influence for vegetables was reported. The group has limited food-preparation skills, but both parents and scouts reported that F&V were available in their homes. Use of goal setting and use of problem-solving techniques were limited. The local scouts' mean F&V intake was 3.2 servings per day. Ethnic differences in F&V consumption were identified in the national data. Based on these results and previous interventions in schools, an overall structure for the intervention was developed to include eight weekly troop sessions and two camping sessions, parent newsletters, seven weekly home badge assignments, and ten comic books.

Annual parallax and a dimming event of a Mira variable star, FV Bootis

NASA Astrophysics Data System (ADS)

Kamezaki, Tatsuya; Nakagawa, Akiharu; Omodaka, Toshihiro; Inoue, Kan-ichiro; Chibueze, James O.; Nagayama, Takumi; Ueno, Yuji; Matsunaga, Noriyuki

2016-10-01

We present the first measurement of the trigonometric parallax of water masers associated with a Mira star, FV Bootis (FV Boo) using VLBI Exploration of Radio Astrometry (VERA). Based on our multi-epoch VERA observations, we derived the parallax to be 0.97 ± 0.06 mas, which corresponds to a distance of 1.03^{+0.07}_{-0.06} kpc. The water masers around FV Boo were spatially distributed over an area of 41 au × 41 au, and their internal motions indicate the presence of an outflow. Using the Kagoshima University 1 m optical/infrared telescope, we determined the period to be 305.6 d and the mean apparent magnitude to be +2.91 mag in the K'-band. On the period-luminosity plane, the obtained period and K'-band magnitude puts FV Boo slightly below the sequence of Miras, possibly due to circumstellar reddening. Combining our photometric data with COBE and 2MASS datasets spanning over 20 years, we found in the near infrared that FV Boo was significantly fainter in 2005 compared with preceding and later phases. Its color, however, did not show a large variation through this change. We infer that the dimming could be caused by an eclipse due to a cloud in a binary system.

LA Sprouts: A 12-Week Gardening, Nutrition, and Cooking Randomized Control Trial Improves Determinants of Dietary Behaviors.

PubMed

Davis, Jaimie N; Martinez, Lauren C; Spruijt-Metz, Donna; Gatto, Nicole M

2016-01-01

To evaluate the effect of an exploratory 12-week nutrition, cooking, and gardening trial (LA Sprouts) on preference for fruit and vegetables (FV); willingness to try FV; identification of FV; self-efficacy to garden, eat, and cook FV; motivation to garden, eat, and cook FV; attitudes toward FV; nutrition and gardening knowledge; and home gardening habits. Randomized controlled trial. Four elementary schools. Three hundred four predominately Hispanic/Latino third- through fifth-grade students were randomized to either the LA Sprouts group (n = 167 students) or control group (n = 137 students). Twelve-week after-school nutrition, cooking, and gardening intervention. Determinants of dietary behavior as measured by questionnaire at baseline and postintervention. Analyses of covariance. After the 12-week program, compared with controls, LA Sprouts participants improved scores for identification of vegetables (+11% vs +5%; P = .001) and nutrition and gardening knowledge (+14.5% vs -5.0%; P = .003), and were more likely to garden at home (+7.5% vs -4.4%; P = .003). The LA Sprouts program positively affected a number of determinants of dietary behaviors that suggest possible mechanisms by which gardening and nutrition education act to improve dietary intake and health outcomes. Copyright © 2016 Society for Nutrition Education and Behavior. Published by Elsevier Inc. All rights reserved.

Relationship between the changes in blood flow and volume in the finger during a Braille character discrimination task.

PubMed

Murata, J; Murata, S; Soma, M; Nakae, H; Sato, Y; Kogo, H; Umeki, N

2017-11-01

We hypothesized that skin blood flow (SBF) of fingers are modulated during concentrated finger perception and that the changes in SBF reflect fluctuations in finger volume (FV). The aim of this study, therefore, was examine the relationship between the changes in SBF and FV during Braille reading. We measured SBF of the finger, cutaneous vascular conductance (CVC), FV, and arterial blood pressure during Braille reading performed under blind conditions in thirty healthy subjects. The subjects were instructed to read a flat plate with raised letters (Braille reading) for 15 seconds using their forefinger, and to touch a blank plate as a control for the Braille discrimination procedure. Arterial blood pressure slightly increased during Braille reading but remained unchanged during the touching of the blank plate. SBF, CVC, and FV were reduced during Braille reading (decreased by -26%, -29%, and -0.3 mL/100 mL respectively). Furthermore, a significant relationship was observed between the changes in SBF and FV (r=.613) during Braille reading. These results suggested that SBF of fingers is modulated during concentrated finger perception, and that the variability of blood flow reflects the response in FV. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Effects of vibration on spasticity in individuals with spinal cord injury: a scoping systematic review.

PubMed

Sadeghi, Mahsa; Sawatzky, Bonita

2014-11-01

The objective of this systematic review was to evaluate how whole-body vibration (WBV) or focal vibration (FV) would change spasticity in individuals with spinal cord injury (SCI). A search was conducted of MEDLINE, EMBASE, CINAHL, and PsycINFO electronic databases. A hand search was conducted of the bibliographies of articles and journals relevant to the research question. The inclusion criteria were three or more individuals, 17 yrs or older, with SCI who experience spasticity, and WBV or FV application. The evidence level of all ten identified studies (195 SCI subjects) was low on the basis of Centre for Evidence Based Medicine level of evidence. WBV (n = 1) and FV (n = 9) were applied to assess the effects of vibration on different measures of spasticity in individuals with SCI. FV application resulted in a short-term spasticity reduction lasting for a maximum of 24 hrs. Neurophysiologic measures showed H-reflex inhibition in individuals with SCI after FV application. WBV resulted in a decrease in spasticity lasting for 6-8 days after the last vibration session. WBV and FV might decrease spasticity for a short period, but no evidence-based recommendation can be drawn from the literature to guide rehabilitation medicine clinicians to manage spasticity with vibration application.

Ligand-induced rapid skeletal muscle atrophy in HSA-Fv2E-PERK transgenic mice.

PubMed

Miyake, Masato; Kuroda, Masashi; Kiyonari, Hiroshi; Takehana, Kenji; Hisanaga, Satoshi; Morimoto, Masatoshi; Zhang, Jun; Oyadomari, Miho; Sakaue, Hiroshi; Oyadomari, Seiichi

2017-01-01

Formation of 43S and 48S preinitiation complexes plays an important role in muscle protein synthesis. There is no muscle-wasting mouse model caused by a repressed 43S preinitiation complex assembly. The aim of the present study was to develop a convenient mouse model of skeletal muscle wasting with repressed 43S preinitiation complex assembly. A ligand-activatable PERK derivative Fv2E-PERK causes the phosphorylation of eukaryotic initiation factor 2α (eIF2α), which inhibits 43S preinitiation complex assembly. Thus, muscle atrophic phenotypes, intracellular signaling pathways, and intracellular free amino acid profiles were investigated in human skeletal muscle α-actin (HSA) promoter-driven Fv2E-PERK transgenic (Tg) mice. HSA-Fv2E-PERK Tg mice treated with the artificial dimerizer AP20187 phosphorylates eIF2α in skeletal muscles and leads to severe muscle atrophy within a few days of ligand injection. Muscle atrophy was accompanied by a counter regulatory activation of mTORC1 signaling. Moreover, intracellular free amino acid levels were distinctively altered in the skeletal muscles of HSA-Fv2E-PERK Tg mice. As a novel model of muscle wasting, HSA-Fv2E-PERK Tg mice provide a convenient tool for studying the pathogenesis of muscle loss and for assessing putative therapeutics.

Behavioral correlates of fruit and vegetable intake in Puerto Rico: results from the Health Information National Trends Survey.

PubMed

Colón-López, Vivian; Banerjee, Geetanjoli; Gertz, Alida María; Ortiz, Ana Patricia; Calo, William; Finney-Rutten, Lila J; Colón-Ramos, Uriyoán; Hesse, Bradford W; Tortolero, Guillermo

2013-12-01

A diet high in fruit and vegetables (FV) is associated with a decreased risk for chronic diseases, such as cancer. Limited information exists regarding the factors associated with FV intake in persons living in Puerto Rico. The objective of this study was to examine sociodemographic, behavioral, and health-belief factors associated with dietary habits in Puerto Rico. Secondary data analysis of adults aged 18 years and older from the Puerto Rico Health Information National Trends Survey (HINTS-PR) conducted in 2009. Multivariate logistic regression models were used to identify factors associated with meeting the established recommendations for FV consumption. Only 14.5% of the adults in Puerto Rico met the recommendations for daily FV intake, and the vast majority (90.9%) were unaware of current recommendations. Bivariate analyses demonstrated that being obese, having lower than a high school education, and not knowing the recommendations were significantly associated with not meeting these recommendations. In the multivariate logistic regression analysis, being obese (OR = 3.77; 95% CI = 1.41-10.08) and not being aware of the current dietary recommendations (OR = 9.26; 95% CI = 3.77-22.73) continued to be significantly associated with not meeting the FV intake recommendations. The Puerto Rican population is far from meeting FV consumption recommendations, with prevalence estimates of consumption that are below the US median. Low FV intake might put the population in Puerto Rico at increased risk of developing cancer as well as a number of other chronic diseases that are secondary both to improper nutrition and to obesity.

Production of a soluble single-chain variable fragment antibody against okadaic acid and exploration of its specific binding.

PubMed

He, Kuo; Zhang, Xiuyuan; Wang, Lixia; Du, Xinjun; Wei, Dong

2016-06-15

Okadaic acid is a lipophilic marine algal toxin commonly responsible for diarrhetic shellfish poisoning (DSP). Outbreaks of DSP have been increasing and are of worldwide public health concern; therefore, there is a growing demand for more rapid, reliable, and economical analytical methods for the detection of this toxin. In this study, anti-okadaic acid single-chain variable fragment (scFv) genes were prepared by cloning heavy and light chain genes from hybridoma cells, followed by fusion of the chains via a linker peptide. An scFv-pLIP6/GN recombinant plasmid was constructed and transformed into Escherichia coli for expression, and the target scFv was identified with IC-CLEIA (chemiluminescent enzyme immunoassay). The IC15 was 0.012 ± 0.02 μg/L, and the IC50 was 0.25 ± 0.03 μg/L. The three-dimensional structure of the scFv was simulated with computer modeling, and okadaic acid was docked to the scFv model to obtain a putative structure of the binding complex. Two predicted critical amino acids, Ser32 and Thr187, were then mutated to verify this theoretical model. Both mutants exhibited significant loss of binding activity. These results help us to understand this specific scFv-antigen binding mechanism and provide guidance for affinity maturation of the antibody in vitro. The high-affinity scFv developed here also has potential for okadaic acid toxin detection. Copyright © 2016. Published by Elsevier Inc.

Identification of NADPH oxidase family members associated with cold stress in strawberry.

PubMed

Zhang, Yunting; Li, Yali; He, Yuwei; Hu, Wenjie; Zhang, Yong; Wang, Xiaorong; Tang, Haoru

2018-04-01

NADPH oxidase is encoded by a small gene family (Respiratory burst oxidase homologs, Rbohs ) and plays an important role in regulating various biological processes. However, little information about this gene family is currently available for strawberry. In this study, a total of seven Rboh genes were identified from strawberry through genomewide analysis. Gene structure analysis showed the number of exons ranged from 10 to 23, implying that this variation occurred in FvRboh genes by the insertion and distribution of introns; the order and approximate size of exons were relatively conserved. FvRbohC was predicted to localize to the thylakoid membrane of the chloroplast, while other members were computed to localize to the plasma membrane, indicating different functions. Amino acid sequence alignment, conserved domain, and motif analysis showed that all identified FvRbohs had typical features of plant Rbohs. Phylogenetic analysis of Rbohs from strawberry, grape, Arabidopsis, and rice suggested that the FvRbohs could be divided into five subgroups and showed a closer relationship with those from grape and Arabidopsis than those from rice. The expression patterns of FvRboh genes in root, stem, leaf, flower, and fruit revealed robust tissue specificity. The expression levels of FvRbohA and FvRbohD were quickly induced by cold stress, followed by an increase in NADPH oxidase activity, leading to O2- accumulation and triggering the antioxidant reaction by the transient increases in SOD activity. This suggested these two genes may be involved in cold stress and defense responses in strawberry.

Clinical utility of balloon expulsion test for functional defecation disorders

PubMed Central

2016-01-01

Purpose I investigated the diagnostic accuracy of balloon expulsion test (BET) with various techniques to find out the most appropriate method, and tried to confirm its clinical utility in diagnosing functional defecation disorders (FDD) in constipated patients. Methods Eighty-seven patients constituted the study population. FDD was defined when patients had at least two positive findings in defecography, manometry, and electromyography. BET was done 4 times in each patient with 2 different positions and 2 different volumes. The positions were seated position (SP) and left lateral decubitus position (LDP). The volumes were fixed volume (FV) of 60 mL and individualized volume with which patient felt a constant desire to defecate (CDV). The results of BETs with 4 different settings (LDP-FV, LDP-CDV, SP-FV, and SP-CDV) were statistically compared and analyzed. Results Of 87 patients, 23 patients (26.4%) had at least two positive findings in 3 tests and thus were diagnosed to have FDD. On receiver operating characteristic curve analysis, area under curve was highest in BET with SP-FV. With a cutoff value of 30 seconds, the specificity of BET with SP-FV was 86.0%, sensitivity was 73.9%, negative predictive value was 89.8%, positive predictive value was 65.4%, and accuracy rate was 82.8% for diagnosing FDD. Conclusion SP-FV is the most appropriate method for BET. In this setting, BET has a diagnostic accuracy sufficient to identify constipated patients who do not have FDD. Patients with negative results in BET with SP-FV may not need other onerous tests to exclude FDD. PMID:26878016

Clinical utility of balloon expulsion test for functional defecation disorders.

PubMed

Seong, Moo-Kyung

2016-02-01

I investigated the diagnostic accuracy of balloon expulsion test (BET) with various techniques to find out the most appropriate method, and tried to confirm its clinical utility in diagnosing functional defecation disorders (FDD) in constipated patients. Eighty-seven patients constituted the study population. FDD was defined when patients had at least two positive findings in defecography, manometry, and electromyography. BET was done 4 times in each patient with 2 different positions and 2 different volumes. The positions were seated position (SP) and left lateral decubitus position (LDP). The volumes were fixed volume (FV) of 60 mL and individualized volume with which patient felt a constant desire to defecate (CDV). The results of BETs with 4 different settings (LDP-FV, LDP-CDV, SP-FV, and SP-CDV) were statistically compared and analyzed. Of 87 patients, 23 patients (26.4%) had at least two positive findings in 3 tests and thus were diagnosed to have FDD. On receiver operating characteristic curve analysis, area under curve was highest in BET with SP-FV. With a cutoff value of 30 seconds, the specificity of BET with SP-FV was 86.0%, sensitivity was 73.9%, negative predictive value was 89.8%, positive predictive value was 65.4%, and accuracy rate was 82.8% for diagnosing FDD. SP-FV is the most appropriate method for BET. In this setting, BET has a diagnostic accuracy sufficient to identify constipated patients who do not have FDD. Patients with negative results in BET with SP-FV may not need other onerous tests to exclude FDD.

Autonomous Motivation and Fruit/Vegetable Intake in Parent–Adolescent Dyads

PubMed Central

Dwyer, Laura A.; Bolger, Niall; Laurenceau, Jean-Philippe; Patrick, Heather; Oh, April Y.; Nebeling, Linda C.; Hennessy, Erin

2017-01-01

Introduction Autonomous motivation (motivation to engage in a behavior because of personal choice, interest, or value) is often associated with health behaviors. The present study contributes to research on motivation and eating behaviors by examining: (1) how autonomous motivation is correlated within parent–adolescent dyads; and (2) whether parent- and adolescent-reported autonomous motivation predicts the parent–adolescent correlation in fruit and vegetable (FV) intake frequency. Methods Data were drawn from the Family Life, Activity, Sun, Health, and Eating (FLASHE) Study, a cross-sectional U.S. survey of parent–adolescent dyads led by the National Cancer Institute and fielded between April and October 2014. In 2016, data were analyzed from dyads who had responses on a six-item self-report measure of daily frequency of FV consumption and a two-item self-report measure of autonomous motivation for consuming FVs. Results Parents' and adolescents' reports of autonomous motivation and FV intake frequency were positively correlated. Both parents' and adolescents' autonomous motivation predicted higher levels of their own FV intake frequency and that of their dyad partner (p-values ≤0.001). These effects of autonomous motivation explained 22.6% of the parent–adolescent correlation in FV intake frequency. Actor effects (one's motivation predicting their own FV intake frequency) were stronger than partner effects (one's motivation predicting their partner's FV intake frequency). Conclusions Parent–adolescent similarity in autonomous motivation for healthy eating may contribute to similarity in eating behaviors. Future research should further examine how individual-level health behavior correlates influence health behaviors within dyads. PMID:28526363

'Kids in the Kitchen' impact evaluation: engaging primary school students in preparing fruit and vegetables for their own consumption.

PubMed

Ritchie, Barbara; O'Hara, Lily; Taylor, Jane

2015-08-01

Children's fruit and vegetable (FV) consumption in Australia is below levels recommended for optimal growth, development and health. 'Kids in the Kitchen' is a classroom-based program that engages students in preparing FV. Impact evaluation was conducted with students from Grade 1 (around 6 years old) and Grade 5 (around 11 years old) who participated in the program. A questionnaire was used to collect pre- and post-program data on knowledge, attitudes and consumption of FV, FV preparation skills and environmental supports for FV consumption. A skill audit was also conducted for Grade 1 students. Study participants (n = 118) included 70 Grade 1 and 48 Grade 5 students. There was an increase in the median number of fruits correctly identified (from 14 to 16), tried (from 14 to 16) and liked (from 10.5 to 12; P = 0.0001 for all changes). The median number of vegetables correctly identified increased from 10 to 12 (P = 0.0001), but there was no change in the number of vegetables tried or liked. The proportion of participants who rated their skills in using a knife to prepare FV as 'not really that good' decreased by 15%, from 42% to 27% (P = 0.04). Grade 1 participants' skills in cutting, grating and peeling improved (P = 0.0001 for all changes). If children are involved in the preparation of FV, they are more likely to correctly identify them, try them, like them and eat them. SO WHAT? Primary schools have the potential to contribute to children's nutrition through hands-on food preparation activities.

Development of the behaviorally focused fruits & Veggies--More Matters public health initiative.

PubMed

Pivonka, Elizabeth; Seymour, Jennifer; McKenna, Jeff; Baxter, Suzanne Domel; Williams, Simon

2011-10-01

Due to changing recommendations for fruit and vegetable (F/V) intake, public health and private organizations recognized the need to revise communications and programs that promote F/V consumption. This article describes formative research conducted in 2005 to develop a new identity for the 5 A Day Program, updated to the Fruits & Veggies--More Matters program. The objective was to re-brand the 5 A Day Program's campaign message to be adaptable, sustainable, and compelling, while leveraging the program's existing message dissemination infrastructure. Formative research included interviews with representatives from government, nonprofit organizations, and industry, and consumer perspectives from interviews, focus groups, and an online survey. Public health and private organizations agreed that a compelling emotional benefit was needed to motivate consumers to eat more F/V and that messaging needed to be used consistently among national, state, and local programs. Interviews and focus groups targeted mothers who believed they and their families were getting enough F/V, knew they could eat more, but needed to be convinced why they should do so. The most effective messages appealed to mothers' emotional needs to be responsible, leveraged functional intrinsic values of F/V, did not try to quantify "enough," and focused on small steps. When the Fruits & Veggies-More Matters slogan and graphic were viewed together, the majority (62%) said it increased their interest in eating more F/V. The Fruits & Veggies-More Matters brand offers numerous opportunities for promoting F/V consumption through this public health initiative. Copyright © 2011 American Dietetic Association. Published by Elsevier Inc. All rights reserved.

Autonomous Motivation and Fruit/Vegetable Intake in Parent-Adolescent Dyads.

PubMed

Dwyer, Laura A; Bolger, Niall; Laurenceau, Jean-Philippe; Patrick, Heather; Oh, April Y; Nebeling, Linda C; Hennessy, Erin

2017-06-01

Autonomous motivation (motivation to engage in a behavior because of personal choice, interest, or value) is often associated with health behaviors. The present study contributes to research on motivation and eating behaviors by examining (1) how autonomous motivation is correlated within parent-adolescent dyads and (2) whether parent- and adolescent-reported autonomous motivation predicts the parent-adolescent correlation in fruit and vegetable (FV) intake frequency. Data were drawn from the Family Life, Activity, Sun, Health, and Eating (FLASHE) Study, a cross-sectional U.S. survey of parent-adolescent dyads led by the National Cancer Institute and fielded between April and October 2014. In 2016, data were analyzed from dyads who had responses on a six-item self-report measure of daily frequency of FV consumption and a two-item self-report measure of autonomous motivation for consuming FVs. Parents' and adolescents' reports of autonomous motivation and FV intake frequency were positively correlated. Both parents' and adolescents' autonomous motivation predicted higher levels of their own FV intake frequency and that of their dyad partner (p-values ≤0.001). These effects of autonomous motivation explained 22.6% of the parent-adolescent correlation in FV intake frequency. Actor effects (one's motivation predicting their own FV intake frequency) were stronger than partner effects (one's motivation predicting their partner's FV intake frequency). Parent-adolescent similarity in autonomous motivation for healthy eating may contribute to similarity in eating behaviors. Future research should further examine how individual-level health behavior correlates influence health behaviors within dyads. Published by Elsevier Inc.

Poor creativity in frontotemporal dementia: a window into the neural bases of the creative mind.

PubMed

de Souza, Leonardo Cruz; Volle, Emmanuelle; Bertoux, Maxime; Czernecki, Virginie; Funkiewiez, Aurélie; Allali, Gilles; Leroy, Baptiste; Sarazin, Marie; Habert, Marie-Odile; Dubois, Bruno; Kas, Aurélie; Levy, Richard

2010-11-01

The prefrontal cortex (PFC) supports functions critical for creative thinking. Damage to the PFC is expected to impair creativity. Yet, previous works suggested the emergence of artistic talent in patients with frontotemporal lobar degeneration (FTLD), which was interpreted as increased creativity. We designed a study in patients with frontal variant (fv) of FTLD in order to verify whether: (1) creativity is impaired after frontal degeneration, (2) poor creativity is associated with frontal dysfunctions, and (3) poor creativity is related to hypoperfusion in specific PFC regions. Three groups of subjects were enrolled in the study: fvFTLD patients (n=17), non-demented Parkinson's disease (PD) patients (n=12) and healthy controls (n=17). Participants performed a standardized test of creativity, the Torrance Test of Creative Thinking (TTCT) and tests assessing frontal functions. Brain perfusion was correlated to fvFTLD patients' performance in the TTCT. Patients with fvFTLD were strongly impaired in all dimensions of the TTCT, compared to PD patients and controls. Disinhibited and perseverative responses were observed only in fvFTLD patients, leading to "pseudo-creative" responses. Poor creativity was positively correlated with several frontal tests. Poor creativity was also correlated with prefrontal hypoperfusion, particularly in the frontal pole. Poor creativity is associated with fvFTLD. The results also suggest that the integrity of the PFC (in particular frontopolar) is strongly associated with creative thinking. The emergence of artistic talent in patients with fvFTLD is explained by the release of involuntary behaviors, rather than by the development of creative thinking. Copyright © 2010 Elsevier Ltd. All rights reserved.

Anti-EpCAM scFv gadolinium chelate: a novel targeted MRI contrast agent for imaging of colorectal cancer.

PubMed

Khantasup, Kannika; Saiviroonporn, Pairash; Jarussophon, Suwatchai; Chantima, Warangkana; Dharakul, Tararaj

2018-05-08

The development of targeted contrast agents for magnetic resonance imaging (MRI) facilitates enhanced cancer imaging and more accurate diagnosis. In the present study, a novel contrast agent was developed by conjugating anti-EpCAM humanized scFv with gadolinium chelate to achieve target specificity. The material design strategy involved site-specific conjugation of the chelating agent to scFv. The scFv monomer was linked to maleimide-DTPA via unpaired cysteine at the scFv C-terminus, followed by chelation with gadolinium (Gd). Successful scFv-DTPA conjugation was achieved at 1:10 molar ratio of scFv to maleimide-DTPA at pH 6.5. The developed anti-EpCAM-Gd-DTPA MRI contrast agent was evaluated for cell targeting ability, in vitro serum stability, cell cytotoxicity, relaxivity, and MR contrast enhancement. A high level of targeting efficacy of anti-EpCAM-Gd-DTPA to an EpCAM-overexpressing HT29 colorectal cell was demonstrated by confocal microscopy. Good stability of the contrast agent was obtained and no cytotoxicity was observed in HT29 cells after 48 h incubation with 25-100 µM of Gd. Favorable imaging was obtained using anti-EpCAM-Gd-DTPA, including 1.8-fold enhanced relaxivity compared with Gd-DTPA, and MR contrast enhancement observed after binding to HT29. The potential benefit of this contrast agent for in vivo MR imaging of colorectal cancer, as well as other EpCAM positive cancers, is suggested and warrants further investigation.

[The alpha-herpesviridae in dermatology : Herpes simplex virus types I and II. German version].

PubMed

El Hayderi, L; Rübben, A; Nikkels, A F

2017-03-01

This review on herpes simplex virus type I and type II (HSV-I, HSV-II) summarizes recent developments in clinical manifestations and treatment interventions for primary and recurrent orolabial and genital herpes, as well as those regarding vaccination issues. Among the clinical presentations, the relationship between pyogenic granuloma and chronic HSV-I infection; HSV-related folliculitis; verrucous HSV-I and HSV-II lesions; the role of recurrent HSV-I infection in burning mouth syndrome; HSV-I and HSV-II infection of the periareolar area; zosteriform HSV; the "knife-cut sign"; and the preferential colonization and infection of preexisting dermatoses by HSV-I or HSV-II are discussed. The usual antiviral treatment regimens for primary and recurrent orolabial and genital herpes are compared to short-term and one-day treatment options. New anti-HSV-I and anti-HSV-II agents include amenavir, pritelivir, brincidofovir, valomaciclovir, and FV-100. Therapeutic or preventive vaccination against HSV-I and HSV-II infections still remains a highly desirable treatment aim, which, unfortunately, has no clinically relevant applications to date.

Exploring the remarkable limits of continuum elastic theory to understand the nanomechanics of viruses

NASA Astrophysics Data System (ADS)

Roos, Wouter; Gibbons, Melissa; Klug, William; Wuite, Gijs

2009-03-01

We report nanoindentation experiments by atomic force microscopy on capsids of the Hepatitis B Virus (HBV). HBV is investigated because its capsids can form in either a smaller T=3 or a bigger T=4 configuration, making it an ideal system to test the predictive power of continuum elastic theory to describe nanometre-sized objects. It is shown that for small, consecutive indentations the particles behave reversibly linear and no material fatigue occurs. For larger indentations the particles start to deform non-linearly. The experimental force response fits very well with finite element simulations on coarse grained models of HBV capsids. Furthermore, this also fits with thin shell simulations guided by the F"oppl- von K'arm'an (FvK) number (the dimensionless ratio of stretching and bending stiffness of a thin shell). Both the T=3 and T=4 morphology are very well described by the simulations and the capsid material turns out to have the same Young's modulus, as expected. The presented results demonstrate the surprising strength of continuum elastic theory to describe indentation of viral capsids.

A high-throughput platform for population reformatting and mammalian expression of phage display libraries to enable functional screening as full-length IgG.

PubMed

Xiao, Xiaodong; Douthwaite, Julie A; Chen, Yan; Kemp, Ben; Kidd, Sara; Percival-Alwyn, Jennifer; Smith, Alison; Goode, Kate; Swerdlow, Bonnie; Lowe, David; Wu, Herren; Dall'Acqua, William F; Chowdhury, Partha S

Phage display antibody libraries are a rich resource for discovery of potential therapeutic antibodies. Single-chain variable fragment (scFv) libraries are the most common format due to the efficient display of scFv by phage particles and the ease by which soluble scFv antibodies can be expressed for high-throughput screening. Typically, a cascade of screening and triaging activities are performed, beginning with the assessment of large numbers of E. coli-expressed scFv, and progressing through additional assays with individual reformatting of the most promising scFv to full-length IgG. However, use of high-throughput screening of scFv for the discovery of full-length IgG is not ideal because of the differences between these molecules. Furthermore, the reformatting step represents a bottle neck in the process because each antibody has to be handled individually to preserve the unique VH and VL pairing. These problems could be resolved if populations of scFv could be reformatted to full-length IgG before screening without disrupting the variable region pairing. Here, we describe a novel strategy that allows the reformatting of diverse populations of scFv from phage selections to full-length IgG in a batch format. The reformatting process maintains the diversity and variable region pairing with high fidelity, and the resulted IgG pool enables high-throughput expression of IgG in mammalian cells and cell-based functional screening. The improved process led to the discovery of potent candidates that are comparable or better than those obtained by traditional methods. This strategy should also be readily applicable to Fab-based phage libraries. Our approach, Screening in Product Format (SiPF), represents a substantial improvement in the field of antibody discovery using phage display.

Determinants of inadequate fruit and vegetable consumption amongst Portuguese adults.

PubMed

Oliveira, A; Maia, B; Lopes, C

2014-04-01

A low consumption of fruit and vegetables (F&V) represents a high burden on health. The present study evaluates sociodemographic, lifestyle and anthropometric determinants of an inadequate consumption of F&V (<5 servings per day). Participants were randomly selected within the adult Porto population (n = 2485). Those with normal cognitive function and information on key variables were analysed (n = 2362). Diet was assessed by a validated food frequency questionnaire. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated by logistic regression, after sex stratification and controlling for age, education, marital status, smoking, regular physical exercise and total energy intake. Older women and men had 37% and 67%, respectively, lower odds of inadequate F&V consumption (≥65 versus <40 years: OR = 0.63, 95% CI = 0.42-0.94; OR = 0.33, 95% CI = 0.20-0.56). More educated subjects (>12 versus <5 years) and those engaged in regular physical exercise had approximately 50% lower probability of F&V inadequacy compared to nonpractitioners, and this was similar in both sexes. On the other hand, female and male current smokers had 1.86- and 2.05-fold higher probabilities of having inadequate F&V consumption. Individuals consuming excessive alcohol (women: ≥15 g day(-1) ; men: ≥30 g day(-1) ) presented a two- and four-fold higher probability of having inadequate F&V consumption compared to nondrinkers (OR = 1.95, 95% CI = 1.38-2.77 in women; OR = 4.40, 95% CI = 2.70-7.18 in men). In both sexes, an inadequate consumption of F&V was more frequently found in younger, less educated and less physically active subjects with smoking and drinking habits. Strategies aiming to increase F&V consumption should consider these target groups that present a clustering of unhealthy lifestyles. © 2013 The British Dietetic Association Ltd.

Using three-phase theory-based formative research to explore healthy eating in Australian truck drivers.

PubMed

Vayro, Caitlin; Hamilton, Kyra

2016-03-01

In Australia, fruit and vegetable consumption is lower than recommended while discretionary foods (i.e., foods high in fat, sugar, and salt) are eaten in excess. Long-haul truck drivers are a group at risk of unhealthy eating but have received limited attention in the health literature. We aimed to examine long-haul truck drivers eating decisions in order to develop theory-based and empirically-driven health messages to improve their healthy food choices. Drawing on the Theory of Planned Behavior, three-phased formative research was conducted using self-report surveys. Phase 1 (N = 30, Mage = 39.53, SDage = 10.72) identified modal salient beliefs about fruit and vegetable (FV) intake and limiting discretionary choices (DC). There were nine behavioral and seven normative beliefs elicited for both FV and DC; while nine and five control beliefs were elicited for FV and DC, respectively. Phase 2 (N = 148, Mage = 44.23, SDage = 12.08) adopted a prospective design with one week follow-up to examine the predictors of FV and DC intention and behavior. A variety of behavioral and control beliefs were predictive of FV and DC intention and behavior. Normative beliefs were predictive of FV intention and behavior and DC intention only. Phase 3 (N = 20, Mage = 46.9, SDage = 12.85) elicited the reasons why each belief is held/solutions to negative beliefs, that could be used as health messages. In total, 40 reasons/solutions were identified: 26 for FV and 14 for DC. In summary, we found that specific behavioral, normative and control beliefs influenced FV and DC eating decisions. These results have implications for truck driver's health and provide formative research to inform future interventions to improve the food choices of a unique group who are at risk of unhealthy eating behaviors. Copyright © 2015 Elsevier Ltd. All rights reserved.

Family Violence Exposure and Health Outcomes Among Older African American Women: Do Spirituality and Social Support Play Protective Roles?

PubMed Central

Kaslow, Nadine

2010-01-01

Abstract Background Family violence (FV), spirituality, and social support are salient psychosocial determinants of health. FV is associated with poor health among older African American women. The effect of spirituality and social support levels on the health of older African American women is unknown. Methods To assess the role of spirituality and social support as culturally relevant determinants of health status for older African American women independent of FV levels, we used a cross-sectional observational study. Two hundred twelve African American women, aged ≥50, were interviewed in two urban primary care practices. The measures used were (1) Family Violence Against Older Women (FVOW) scale, (2) Physical and Mental Composite Scores of the Short-Form 8® scale, (3) Medical Outcomes of Social Support survey (MOSS), and (4) Spiritual Well-Being Scale (SWBS). Spearman correlation coefficients estimated to test associations among lifetime FV exposure, spirituality, social support, and health status outcomes and multivariate regression models were used to examine the independent effect of spirituality and social support on physical and mental health status, controlling for FV and significant demographic variables. Results Mean participant age was 63.9 years. Higher spirituality levels were significantly associated with better physical health status after adjusting for FV levels and demographic factors (F = 6.17, p = 0.0001). Similarly, higher levels of spirituality and social support both significantly correlated with better mental health status in the multivariate model (F = 13.45, p < 0.0001) that controlled for lifetime FV levels and demographic factors. Conclusions Spirituality and social support are two potentially modifiable determinants of health for older African American women. Culturally appropriate mechanisms to enhance social support and spirituality levels need to be explored as potential inteventions to improve the health of those African American women who have been exposed to FV. PMID:20831432

Expression and purification of a novel therapeutic single-chain variable fragment antibody against BNP from inclusion bodies of Escherichia coli.

PubMed

Bu, Dawei; Zhou, Yuwei; Tang, Jian; Jing, Fang; Zhang, Wei

2013-12-01

Abnormal brain natriuretic peptide (BNP) secretion is regarded as the dominating mechanism of cerebral salt wasting syndrome (CSW), which results from a renal loss of sodium and water during intracranial disease leading to hyponatremia. Scale preparation of therapeutic single-chain variable fragment (scFv) that can neutralize elevated circulating BNP may have potential value for clinical use. In this report, we used a recently isolated humanized anti-BNP scFv fragment (3C1) as model antibody (Ab) to evaluate the potential of scale production of this therapeutic protein. The truncated gene encoding for scFv fragment cloned in pET22b (+) was mainly overexpressed as inclusion bodies in Escherichia coli (E. coli) Rosetta (DE3) pLysS cells. The insoluble fragment was solubilized and purified by Ni-NTA agarose resin under denaturation conditions, and recovered via an effective refolding buffer containing 50 mM Tris-HCl, pH 8.0, 0.15 M NaCl, 1 mM EDTA, 0.5 M arginine, 2 mM GSH, 1 mM GSSG, and 5% glycerol. The refolded scFv fragment was concentrated by PEG20000, and dialyzed in PBS (containing 5% glycerol, pH 7.4). The final yield was approximately 10.2 mg active scFv fragment per liter of culture (3.4 g wet weight cells). The scFv fragment was more than 95% pure assessed by SDS-PAGE assay. Recombinant scFv fragment with His tag displayed its immunoreactivity with anti-His tag Ab by western blotting. ELISA showed the scFv fragment specifically bound to BNP, and it displayed similar activity as the traditional anti-BNP monoclonal Ab (mAb). Thus, the current strategy allows convenient small-scale production of this therapeutic protein. Copyright © 2013 Elsevier Inc. All rights reserved.

Increasing Fruit and Vegetable Intake among Children and Youth through Gardening-Based Interventions: A Systematic Review.

PubMed

Savoie-Roskos, Mateja R; Wengreen, Heidi; Durward, Carrie

2017-02-01

Although there are numerous health benefits associated with eating fruit and vegetables (F/V), few children are consuming recommended amounts. Gardening interventions have been implemented in various settings in an effort to increase children's F/V consumption by expanding knowledge, exposure, and preferences for a variety of F/V. The purpose of this review was to identify the effectiveness of gardening interventions that have been implemented to increase F/V consumption among children. A systematic review was conducted using four electronic databases: Web of Science, PubMed, Scopus, and the Cumulative Index to Nursing and Allied Health Literature. English language studies conducted in developed countries between January 2005 and October 2015 were included in this review. Included studies measured F/V consumption among children aged 2 to 15 years before and after implementation of a gardening intervention in a school, community, or afterschool setting. All study designs were included in this review. A total of 891 articles were identified through database searching and cross-referencing. After removing duplicates, 650 articles remained and were screened using inclusion and exclusion criteria. Twenty-seven full-text articles were analyzed and 14 articles were included in this review. Of the 14 articles reviewed, 10 articles found statistically significant increases in fruit or vegetable consumption among participants after implementation of a gardening intervention. However, many studies were limited by the use of convenience samples, small sample sizes, and self-reported measurements of F/V consumption. Although the evidence is mixed and fraught with limitations, most studies suggest a small but positive influence of gardening interventions on children's F/V intake. Future studies that include control groups, randomized designs, and assessments of F/V consumption over at least 1 year are needed to advance the literature on this topic. Copyright © 2017 Academy of Nutrition and Dietetics. Published by Elsevier Inc. All rights reserved.

Cell growth inhibition and apoptosis in breast cancer cells induced by anti-FZD7 scFvs: involvement of bioinformatics-based design of novel epitopes.

PubMed

Zarei, Neda; Fazeli, Mehdi; Mohammadi, Mozafar; Nejatollahi, Foroogh

2018-06-01

FZD7 has a critical role as a surface receptor of Wnt/β-catenin signaling in cancer cells. Suppressing Wnt signaling through blocking FZD7 is shown to decrease cell viability, metastasis and invasion. Bioinformatic methods have been a powerful tool in epitope designing studies. Small size, high affinity and human origin of scFv antibodies have provided unique advantages for these recombinant antibodies. Two epitopes from extracellular domain of FZD7 were designed using bioinformatic methods. Specific anti-FZD7 scFvs were selected against these epitopes through panning process. The specificity of the scFvs was assessed by phage ELISA and the ability to bind to FZD7 expressing cell line (MDA-MB-231) was determined by flowcytometry. Antiproliferative and apoptotic effects of the scFvs were evaluated by MTT and Annexin V/PI assays. The effects of selected scFvs on expression level of Surivin, c-Myc and Dvl genes were also evaluated by real-time PCR. Results demonstrated selection of two specific scFvs (scFv-I and scFv-II) with frequencies of 35 and 20%. Both antibodies bound to the corresponding peptides and cell surface receptors as shown by phage ELISA and flowcytometry, respectively. The scFvs inhibited cell growth of MDA-MB-231 cells significantly as compared to untreated cells. Growth inhibition of 58.6 and 53.1% were detected for scFv-I and scFv-II, respectively. No significant growth inhibition was detected for SKBR-3 negative control cells. The scFvs induced apoptotic effects in the MDA-MB-231 treated cells after 48 h, which were 81.6 and 74.9% for scFv-I and scFv-II, respectively. Downregulation of Surivin, c-Myc and Dvl genes were also shown after 48h treatment of cells with either of scFvs (59.3-93.8%). ScFv-I showed significant higher antiproliferative and apoptotic effects than scFv-II. Bioinformatic methods could effectively select potential epitopes of FZD7 protein and suggest that epitope designing by bioinformatic methods could contribute to the selection of key antigens for cancer immunotherapy. The selected scFvs, especially scFv-I, with high antiproliferative and apoptotic effects could be considered as effective agents for immunotherapy of cancers expressing FZD7 receptor including triple negative breast cancer.

Validity and reliability of a brief self-reported questionnaire assessing fruit and vegetable consumption among pregnant women.

PubMed

Vézina-Im, Lydi-Anne; Godin, Gaston; Couillard, Charles; Perron, Julie; Lemieux, Simone; Robitaille, Julie

2016-09-15

Short instruments measuring frequency of specific foods, such as fruit and vegetable (FV), are increasingly used in interventions. The objective of the study was to verify the validity and test-retest reliability of such an instrument among pregnant women. Pregnant women from the region of Quebec City, Quebec, Canada, were recruited through e-mails sent to female students and employees of the local university from October 2014 to April 2015. To assess the validity of the fruit and vegetable questionnaire (FVQ) developed by Godin et al. (Can J Public Health 99: 494-498, 2008), pregnant women were asked in a first mailing to complete the FVQ assessing FV intake over the past 7 days and a 3-day estimated food record. A subsample (n = 33) also gave a fasting blood sample and completed a validated semi-quantitative FFQ administered by a trained registered dietitian during a visit at the research center. FV intakes for all instruments were calculated in terms of servings of FV based on Canada's Food Guide definition of a serving of fruit or vegetable. In order to assess its test-retest reliability, respondents were asked to complete the FVQ 14 days later in a second mailing. Forty-eight pregnant women from all three trimesters completed the questionnaires in the first mailing. FV intake assessed using the FVQ was correlated to FV consumption measured using the food record (r = 0.34, p = 0.0180) and the FFQ (r = 0.61, p = 0.0002). Results were similar when controlling for energy intake and the experience of nausea in the past month. Only β-cryptoxanthin was significantly correlated to FV intake assessed by the FFQ when adjusted for the presence of nausea (r = 0.35, p = 0.0471). Data on the test-retest reliability was available for 44 women and the intra-class coefficient for the FVQ was 0.72 at a mean 28-day interval. The FVQ has acceptable validity and test-retest reliability values, but seems to underestimate FV servings in pregnant women. It represents an interesting alternative for researchers or clinicians interested in estimating quickly FV intake among pregnant women, such as in large trials or during prenatal visits. The FVQ should however be coupled with other self-reported measures, such as a food record, for assessing precise individual FV intake.

Erythroblast Transformation by the Friend Spleen Focus-Forming Virus Is Associated with a Block in Erythropoietin-Induced STAT1 Phosphorylation and DNA Binding and Correlates with High Expression of the Hematopoietic Phosphatase SHP-1

PubMed Central

Nishigaki, Kazuo; Hanson, Charlotte; Ohashi, Takashi; Spadaccini, Angelo; Ruscetti, Sandra

2006-01-01

Infection of mice with Friend spleen focus-forming virus (SFFV) results in a multistage erythroleukemia. In the first stage, the SFFV envelope glycoprotein interacts with the erythropoietin receptor and a short form of the receptor tyrosine kinase sf-Stk, resulting in constitutive activation of signal transducing molecules and the development of erythropoietin (Epo)-independent erythroid hyperplasia and polycythemia. The second stage results from the outgrowth of a rare virus-infected erythroid cell that expresses nonphysiological levels of the myeloid transcription factor PU.1. These cells exhibit a differentiation block and can be grown as murine erythroleukemia (MEL) cell lines. In this study, we examined SFFV MEL cells to determine whether their transformed phenotype was associated with a block in the activation of any Epo signal-transducing molecules. Our studies indicate that Epo- or SFFV-induced activation of STAT1/3 DNA binding activity is blocked in SFFV MEL cells. The block is at the level of tyrosine phosphorylation of STAT1, although Jak2 phosphorylation is not blocked in these cells. In contrast to Epo, alpha interferon can induce STAT1 phosphorylation and DNA binding in SFFV MEL cells. The SFFV-transformed cells were shown to express elevated levels of the hematopoietic phosphatase SHP-1, and treatment of the cells with a phosphatase inhibitor restored STAT1 tyrosine phosphorylation. MEL cells derived from Friend murine leukemia virus (MuLV) or ME26 MuLV-infected mice, which do not express PU.1, express lower levels of SHP-1 and are not blocked in STAT1/3 DNA-binding activity. Our studies suggest that SFFV-infected erythroid cells become transformed when differentiation signals activated by STAT1/3 are blocked due to high SHP-1 levels induced by inappropriate expression of the PU.1 protein. PMID:16731906

XTHs from Fragaria vesca: genomic structure and transcriptomic analysis in ripening fruit and other tissues.

PubMed

Opazo, María Cecilia; Lizana, Rodrigo; Stappung, Yazmina; Davis, Thomas M; Herrera, Raúl; Moya-León, María Alejandra

2017-11-07

Fragaria vesca or 'woodland strawberry' has emerged as an attractive model for the study of ripening of non-climacteric fruit. It has several advantages, such as its small genome and its diploidy. The recent availability of the complete sequence of its genome opens the possibility for further analysis and its use as a reference species. Fruit softening is a physiological event and involves many biochemical changes that take place at the final stages of fruit development; among them, the remodeling of cell walls by the action of a set of enzymes. Xyloglucan endotransglycosylase/hydrolase (XTH) is a cell wall-associated enzyme, which is encoded by a multigene family. Its action modifies the structure of xyloglucans, a diverse group of polysaccharides that crosslink with cellulose microfibrills, affecting therefore the functional structure of the cell wall. The aim of this work is to identify the XTH-encoding genes present in F. vesca and to determine its transcription level in ripening fruit. The search resulted in identification of 26 XTH-encoding genes named as FvXTHs. Genetic structure and phylogenetic analyses were performed allowing the classification of FvXTH genes into three phylogenetic groups: 17 in group I/II, 2 in group IIIA and 4 in group IIIB. Two sequences were included into the ancestral group. Through a comparative analysis, characteristic structural protein domains were found in FvXTH protein sequences. In complement, expression analyses of FvXTHs by qPCR were performed in fruit at different developmental and ripening stages, as well as, in other tissues. The results showed a diverse expression pattern of FvXTHs in several tissues, although most of them are highly expressed in roots. Their expression patterns are not related to their respective phylogenetic groups. In addition, most FvXTHs are expressed in ripe fruit, and interestingly, some of them (FvXTH 18 and 20, belonging to phylogenic group I/II, and FvXTH 25 and 26 to group IIIB) display an increasing expression pattern as the fruit ripens. A discrete group of FvXTHs (18, 20, 25 and 26) increases their expression during softening of F. vesca fruit, and could take part in cell wall remodeling required for softening in collaboration with other cell wall degrading enzymes.

Comparing effectiveness of mass media campaigns with price reductions targeting fruit and vegetable intake on US cardiovascular disease mortality and race disparities.

PubMed

Pearson-Stuttard, Jonathan; Bandosz, Piotr; Rehm, Colin D; Afshin, Ashkan; Peñalvo, Jose L; Whitsel, Laurie; Danaei, Goodarz; Micha, Renata; Gaziano, Tom; Lloyd-Williams, Ffion; Capewell, Simon; Mozaffarian, Dariush; O'Flaherty, Martin

2017-07-01

Background: A low intake of fruits and vegetables (F&Vs) is a major risk factor for cardiovascular disease (CVD) in the United States. Both mass media campaigns (MMCs) and economic incentives may increase F&V consumption. Few data exist on their comparative effectiveness. Objective: We estimated CVD mortality reductions potentially achievable by price reductions and MMC interventions targeting F&V intake in the US population. Design: We developed a US IMPACT Food Policy Model to compare 3 policies targeting F&V intake across US adults from 2015 to 2030: national MMCs and national F&V price reductions of 10% and 30%. We accounted for differences in baseline diets, CVD rates, MMC coverage, MMC duration, and declining effects over time. Outcomes included cumulative CVD (coronary heart disease and stroke) deaths prevented or postponed and life-years gained (LYGs) over the study period, stratified by age, sex, and race. Results: A 1-y MMC in 2015 would increase the average national F&V consumption by 7% for 1 y and prevent ∼18,600 CVD deaths (95% CI: 17,600, 19,500), gaining ∼280,100 LYGs by 2030. With a 15-y MMC, increased F&V consumption would be sustained, yielding a 3-fold larger reduction (56,100; 95% CI: 52,400, 57,700) in CVD deaths. In comparison, a 10% decrease in F&V prices would increase F&V consumption by ∼14%. This would prevent ∼153,300 deaths (95% CI: 146,400, 159,200), gaining ∼2.51 million LYGs. For a 30% price decrease, resulting in a 42% increase in F&V consumption, corresponding values would be 451,900 CVD deaths prevented or postponed (95% CI: 433,100, 467,500) and 7.3 million LYGs gained. Effects were similar by sex, with a smaller proportional effect and larger absolute effects at older ages. A 1-y MMC would be 35% less effective in preventing CVD deaths in non-Hispanic blacks than in whites. In comparison, price-reduction policies would have equitable proportional effects. Conclusion: Both national MMCs and price-reduction policies could reduce US CVD mortality, with price reduction being more powerful and sustainable. © 2017 American Society for Nutrition.

[Targeted detecting HER2 expression with recombinant anti HER2 ScFv-GFP fusion antibody].

PubMed

Gao, Guohui; Chen, Chong; Yang, Yanmei; Yang, Han; Wang, Jindan; Zheng, Yi; Huang, Qidi; Hu, Xiaoqu

2012-08-01

To verify the reliability of targeted detecting HER2 positive cancer cells and clinical pathological tissue specimens with a recombinant anti HER2 single chain antibody in single chain Fv fragment (scFv) format, we have constructed the fusion variable regions of the ScFv specific for HER2/neu. labeled a green-fluorescent protein(GFP). The humanized recombinant Anti HER2 ScFv-GFP gene was inserted into pFast Bac HT A, and expressed in insect cells sf9. Then the recombinant fusion protein Anti HER2 ScFv-GFP was properly purified with Ni2+-NTA affinity chromatography from the infected sf9 cells used to test the specificity of the fusion antibody for HER2 positive cancer cells. Firstly, the purified antibody incubated with HER2 positive breast cancer cells SKBR3, BT474 and HER2 negative breast cancer cells MCF7 for 12 h/24 h/48 h at 37 degrees C, in order to confirm targeted detecting HER2 positive breast cancer cells by Laser Confocal Microscopy. Furthermore, the same clinical pathological tissue samples were assessed by immunohistochemistry (IHC) and the fusion antibody Anti HER2 ScFv-GFP in the meanwhile. The data obtained indicated that the recombinant eukaryotic expression plasmid pFast Bac HT A/Anti HER2 ScFv-GFP was constructed successfully In addition, obvious green fluorescent was observed in insect cells sf9. When the purified fusion antibody was incubated with different cancer cells, much more green fluorescent was observed on the surface of the HER2 positive cancer cells SKBR3 and BT474. In contrast, no green fluorescent on the surface of the HER2 negative cancer cells MCF7 was detected. The concentration of the purified fusion antibody was 115.5 microg/mL, of which protein relative molecular weight was 60 kDa. The analysis showed the purity was about 97% and the titer was about 1:64. The detection results of IHC and fusion antibody testing indicated the conformity. In summary, the study showed that the new fusion antibody Anti HER2 ScFv-GFP can test HER2 positive cancer cells, indicating a potential candidate method for clinical HER2 positive specimens detection.

Applications and Improvement of a Coupled, Global and Cloud-Resolving Modeling System

NASA Technical Reports Server (NTRS)

Tao, W.-K.; Chern, J.; Atlas, R.

2005-01-01

Recently Grabowski (2001) and Khairoutdinov and Randall (2001) have proposed the use of 2D CFWs as a "super parameterization" [or multi-scale modeling framework (MMF)] to represent cloud processes within atmospheric general circulation models (GCMs). In the MMF, a fine-resolution 2D CRM takes the place of the single-column parameterization used in conventional GCMs. A prototype Goddard MMF based on the 2D Goddard Cumulus Ensemble (GCE) model and the Goddard finite volume general circulation model (fvGCM) is now being developed. The prototype includes the fvGCM run at 2.50 x 20 horizontal resolution with 32 vertical layers from the surface to 1 mb and the 2D (x-z) GCE using 64 horizontal and 32 vertical grid points with 4 km horizontal resolution and a cyclic lateral boundary. The time step for the 2D GCE would be 15 seconds, and the fvGCM-GCE coupling frequency would be 30 minutes (i.e. the fvGCM physical time step). We have successfully developed an fvGCM-GCE coupler for this prototype. Because the vertical coordinate of the fvGCM (a terrain-following floating Lagrangian coordinate) is different from that of the GCE (a z coordinate), vertical interpolations between the two coordinates are needed in the coupler. In interpolating fields from the GCE to fvGCM, we use an existing fvGCM finite- volume piecewise parabolic mapping (PPM) algorithm, which conserves the mass, momentum, and total energy. A new finite-volume PPM algorithm, which conserves the mass, momentum and moist static energy in the z coordinate, is being developed for interpolating fields from the fvGCM to the GCE. In the meeting, we will discuss the major differences between the two MMFs (i.e., the CSU MMF and the Goddard MMF). We will also present performance and critical issues related to the MMFs. In addition, we will present multi-dimensional cloud datasets (i.e., a cloud data library) generated by the Goddard MMF that will be provided to the global modeling community to help improve the representation and performance of moist processes in climate models and to improve our understanding of cloud processes globally (the software tools needed to produce cloud statistics and to identify various types of clouds and cloud systems from both high-resolution satellite and model data will be also presented).

Enhancing the Oncolytic Activity of CD133-Targeted Measles Virus: Receptor Extension or Chimerism with Vesicular Stomatitis Virus Are Most Effective

PubMed Central

Kleinlützum, Dina; Hanauer, Julia D. S.; Muik, Alexander; Hanschmann, Kay-Martin; Kays, Sarah-Katharina; Ayala-Breton, Camilo; Peng, Kah-Whye; Mühlebach, Michael D.; Abel, Tobias; Buchholz, Christian J.

2017-01-01

Therapy resistance and tumor recurrence are often linked to a small refractory and highly tumorigenic subpopulation of neoplastic cells, known as cancer stem cells (CSCs). A putative marker of CSCs is CD133 (prominin-1). We have previously described a CD133-targeted oncolytic measles virus (MV-CD133) as a promising approach to specifically eliminate CD133-positive tumor cells. Selectivity was introduced at the level of cell entry by an engineered MV hemagglutinin (H). The H protein was blinded for its native receptors and displayed a CD133-specific single-chain antibody fragment (scFv) as targeting domain. Interestingly, MV-CD133 was more active in killing CD133-positive tumors than the unmodified MV-NSe despite being highly selective for its target cells. To further enhance the antitumoral activity of MV-CD133, we here pursued arming technologies, receptor extension, and chimeras between MV-CD133 and vesicular stomatitis virus (VSV). All newly generated viruses including VSV-CD133 were highly selective in eliminating CD133-positive cells. MV-CD46/CD133 killed in addition CD133-negative cells being positive for the MV receptors. In an orthotopic glioma model, MV-CD46/CD133 and MVSCD-CD133, which encodes the super cytosine deaminase, were most effective. Notably, VSV-CD133 caused fatal neurotoxicity in this tumor model. Use of CD133 as receptor could be excluded as being causative. In a subcutaneous tumor model of hepatocellular cancer, VSV-CD133 revealed the most potent oncolytic activity and also significantly prolonged survival of the mice when injected intravenously. Compared to MV-CD133, VSV-CD133 infected a more than 104-fold larger area of the tumor within the same time period. Our data not only suggest new concepts and approaches toward enhancing the oncolytic activity of CD133-targeted oncolytic viruses but also raise awareness about careful toxicity testing of novel virus types. PMID:28695108

Population parameters for resistance to Fusarium graminearum and Fusarium verticillioides ear rot among large sets of early, mid-late and late maturing European maize (Zea mays L.) inbred lines.

PubMed

Löffler, Martin; Kessel, Bettina; Ouzunova, Milena; Miedaner, Thomas

2010-03-01

Infection of maize ears with Fusarium graminearum (FG) and Fusarium verticillioides (FV) reduces yield and quality by mycotoxin contamination. Breeding and growing varieties resistant to both Fusarium spp. is the best alternative to minimize problems. The objectives of our study were to draw conclusions on breeding for ear rot resistance by estimating variance components, heritabilities and correlations between resistances to FV and FG severity and to investigate different inoculation methods. In 2007 and 2008, three maturity groups (early, mid-late, late) each comprising about 150 inbred lines were tested in Germany, France, Italy, and Hungary according to their maturity group. They were silk channel inoculated by FG (early) and FV (all groups). In the late maturity group, additionally kernel inoculation was applied in a separate trial. The percentage of mycelium coverage on the ear was rated at harvest (0-100%). Significant (P < 0.01) genotypic variances of ear rot severity were found in all groups. Inoculation was superior to natural infection because of higher disease severities and heritabilities. In early maturing flints and dents, FG caused significantly (P < 0.01) higher ear rot severity than FV (61.7 and 55.1% FG vs. 18.2 and 11.1% FV ear rot severity, respectively). FV inoculation in Southern Europe (mid-late, late) resulted in similar means between 10.3 and 14.0%. Selection is complicated by significant (P < 0.01) genotype x environment interactions. Correlation between FG and FV severity was moderate in flints and dents (r = 0.59 and 0.49, respectively) but lines resistant to both fungi exist. We conclude that chances for selecting improved European elite maize material within the existing germplasms is promising by multi-environmental inoculation trials.

Behavioral Correlates of Fruit and Vegetable Intake in Puerto Rico: Results from the Health Information National Trends Survey

PubMed Central

Colón-López, Vivian; Banerjee, Geetanjoli; Gertz, Alida María; Ortiz, Ana Patricia; Calo, William; Finney-Rutten, Lila J.; Colón-Ramos, Uriyoán; Hesse, Bradford W.; Tortolero, Guillermo

2016-01-01

Objective A diet high in fruit and vegetables (FV) is associated with a decreased risk for chronic diseases, such as cancer. Limited information exists regarding the factors associated with FV intake in persons living in Puerto Rico. The objective of this study was to examine sociodemographic, behavioral, and health-belief factors associated with dietary habits in Puerto Rico. Methods Secondary data analysis of adults aged 18 years and older from the Puerto Rico Health Information National Trends Survey (HINTS-PR) conducted in 2009. Multivariate logistic regression models were used to identify factors associated with meeting the established recommendations for FV consumption. Results Only 14.5% of the adults in Puerto Rico met the recommendations for daily FV intake, and the vast majority (90.9%) were unaware of current recommendations. Bivariate analyses demonstrated that being obese, having lower than a high school education, and not knowing the recommendations were significantly associated with not meeting these recommendations. In the multivariate logistic regression analysis, being obese (OR = 3.77; 95% CI = 1.41-10.08) and not being aware of the current dietary recommendations (OR = 9.26; 95% CI = 3.77-22.73) continued to be significantly associated with not meeting the FV intake recommendations. Conclusion The Puerto Rican population is far from meeting FV consumption recommendations, with prevalence estimates of consumption that are below the US median. Low FV intake might put the population in Puerto Rico at increased risk of developing cancer as well as a number of other chronic diseases that are secondary both to improper nutrition and to obesity. PMID:24397217

Efficacy of a store-based environmental change intervention compared with a delayed treatment control condition on store customers’ intake of fruits and vegetables

PubMed Central

Ayala, Guadalupe X; Baquero, Barbara; Laraia, Barbara A; Ji, Ming; Linnan, Laura

2013-01-01

Objective The present store-based intervention was designed to promote sales of fruits and vegetables (F&V) to increase intake among store customers – specifically customers of tiendas, small-to-medium-sized Latino food stores. Design Four tiendas were randomized to a 2-month environmental change intervention or a delayed treatment control condition. Employees and managers were trained to promote F&V sales, including how to implement a food marketing campaign and installing store equipment to promote fresh fruits and vegetables. The primary outcome was self-reported daily intake of F&V among a convenience sample of customers (at least forty per store) collected at baseline prior to randomization and then 4 months later. In addition, changes in availability of F&V in the tiendas, using unobtrusive observational methods, provided evidence of intervention fidelity. Setting Tiendas in central North Carolina. Subjects Participants included 179 customers who were recent immigrants from Mexico and Central America. Results A group-by-time interaction approached significance on daily servings of F&V; intervention customers reported an increase in F&V intake over time and as a function of the intervention (P≤0·06). Unexpectedly, self-efficacy for consuming more fruits (P≤0·01) and more vegetables (P≤0·06) decreased. In our store-level analyses, a group-by-time interaction was observed for availability of fresh and canned vegetables; the intervention increased availability of vegetables but not fruit. Conclusions Environmental change strategies to promote healthy eating are needed given the rates of obesity and diabetes in the Latino population. A store-based intervention was moderately effective at increasing customers’ reported F&V intake. Such strategies can have a public health impact on underserved populations. PMID:23561842

Parenting practices are associated with fruit and vegetable consumption in pre-school children

PubMed Central

O’Connor, Teresia M; Hughes, Sheryl O; Watson, Kathy B; Baranowski, Tom; Nicklas, Theresa A; Fisher, Jennie O; Beltran, Alicia; Baranowski, Janice C; Qu, Haiyan; Shewchuk, Richard M

2009-01-01

Objective Parents may influence children’s fruit and vegetable (F&V) consumption in many ways, but research has focused primarily on counterproductive parenting practices, such as restriction and pressure to eat. The present study aimed to assess the association of diverse parenting practices to promote F&V and its consumption among pre-school children. Design An exploratory analysis was performed on cross-sectional data from 755 Head Start pre-school children and their parents collected in 2004–5. Data included parent practices to facilitate child F&V consumption (grouped into five categories); parent-reported dietary intake of their child over 3 d; and a number of potential correlates. K-means cluster analysis assigned parents to groups with similar use of the food parenting practice categories. Stepwise linear regression analyses investigated the association of parent clusters with children’s consumption of F&V, after controlling for potential confounding factors. Results A three-cluster solution provided the best fit (R2 = 0 62), with substantial differences in the use of parenting practices. The clusters were labelled Indiscriminate Food Parenting, Non-directive Food Parenting and Low-involved Food Parenting. Non-directive parents extensively used enhanced availability and teachable moments’ practices, but less firm discipline practices than the other clusters, and were significantly associated with child F&V intake (standardized β = 0·09, P < 0·1; final model R2 = 0·17) after controlling for confounders, including parental feeding styles. Conclusions Parents use a variety of parenting practices, beyond pressuring to eat and restrictive practices, to promote F&V intake in their young child. Evaluating the use of combinations of practices may provide a better understanding of parental influences on children’s F&V intake. PMID:19490734

Modification and identification of a vector for making a large phage antibody library.

PubMed

Zhang, Guo-min; Chen, Yü-ping; Guan, Yuan-zhi; Wang, Yan; An, Yun-qing

2007-11-20

The large phage antibody library is used to obtain high-affinity human antibody, and the Loxp/cre site-specific recombination system is a potential method for constructing a large phage antibody library. In the present study, a phage antibody library vector pDF was reconstructed to construct diabody more quickly and conveniently without injury to homologous recombination and the expression function of the vector and thus to integrate construction of the large phage antibody library with the preparation of diabodies. scFv was obtained by overlap polymerase chain reaction (PCR) amplification with the newly designed VL and VH extension primers. loxp511 was flanked by VL and VH and the endonuclease ACC III encoding sequences were introduced on both sides of loxp511. scFv was cloned into the vector pDF to obtain the vector pDscFv. The vector expression function was identified and the feasibility of diabody preparation was evaluated. A large phage antibody library was constructed in pDscFv. Several antigens were used to screen the antibody library and the quality of the antibody library was evaluated. The phage antibody library expression vector pDscFv was successfully constructed and confirmed to express functional scFv. The large phage antibody library constructed using this vector was of high diversity. Screening of the library on 6 antigens confirmed the generation of specific antibodies to these antigens. Two antibodies were subjected to enzymatic digestion and were prepared into diabody with functional expression. The reconstructed vector pDscFv retains its recombination capability and expression function and can be used to construct large phage antibody libraries. It can be used as a convenient and quick method for preparing diabodies after simple enzymatic digestion, which facilitates clinical trials and application of antibody therapy.

Environmental influences on fruit and vegetable intake: Results from a path analytic model

PubMed Central

Liese, Angela D.; Bell, Bethany A.; Barnes, Timothy L.; Colabianchi, Natalie; Hibbert, James D.; Blake, Christine E.; Freedman, Darcy A.

2014-01-01

Objective Fruit and vegetable intake (F&V) is influenced by behavioral and environmental factors, but these have rarely been assessed simultaneously. We aimed to quantify the relative influence of supermarket availability, perceptions of the food environment, and shopping behavior on F&V intake. Design A cross-sectional study. Setting Eight-counties in South Carolina, USA, with verified locations of all supermarkets. Subjects A telephone survey of 831 household food shoppers ascertained F&V intake with a 17-item screener, primary food store location, shopping frequency, perceptions of healthy food availability, and calculated GIS-based supermarket availability. Path analysis was conducted. We report standardized beta coefficients on paths significant at the 0.05 level. Results Frequency of grocery shopping at primary food store (β=0.11) was the only factor exerting an independent, statistically significant direct effect on F&V intake. Supermarket availability was significantly associated with distance to food store (β=-0.24) and shopping frequency (β=0.10). Increased supermarket availability was significantly and positively related to perceived healthy food availability in the neighborhood (β=0.18) and ease of shopping access (β=0.09). Collectively considering all model paths linked to perceived availability of healthy foods, this measure was the only other factor to have a significant total effect on F&V intake. Conclusions While the majority of literature to date has suggested an independent and important role of supermarket availability for F&V intake, our study found only indirect effects of supermarket availability and suggests that food shopping frequency and perceptions of healthy food availability are two integral components of a network of influences on F&V intake. PMID:24192274

Efficacy of a store-based environmental change intervention compared with a delayed treatment control condition on store customers' intake of fruits and vegetables.

PubMed

Ayala, Guadalupe X; Baquero, Barbara; Laraia, Barbara A; Ji, Ming; Linnan, Laura

2013-11-01

The present store-based intervention was designed to promote sales of fruits and vegetables (F&V) to increase intake among store customers--specifically customers of tiendas, small-to-medium-sized Latino food stores. Four tiendas were randomized to a 2-month environmental change intervention or a delayed treatment control condition. Employees and managers were trained to promote F&V sales, including how to implement a food marketing campaign and installing store equipment to promote fresh fruits and vegetables. The primary outcome was self-reported daily intake of F&V among a convenience sample of customers (at least forty per store) collected at baseline prior to randomization and then 4 months later. In addition, changes in availability of F&V in the tiendas, using unobtrusive observational methods, provided evidence of intervention fidelity. Tiendas in central North Carolina. Participants included 179 customers who were recent immigrants from Mexico and Central America. A group-by-time interaction approached significance on daily servings of F&V; intervention customers reported an increase in F&V intake over time and as a function of the intervention (P < or = 0.06). Unexpectedly, self-efficacy for consuming more fruits (P < or = 0.01) and more vegetables (P < or = 0.06) decreased. In our store-level analyses, a group-by-time interaction was observed for availability of fresh and canned vegetables; the intervention increased availability of vegetables but not fruit. Environmental change strategies to promote healthy eating are needed given the rates of obesity and diabetes in the Latino population. A store-based intervention was moderately effective at increasing customers’ reported F&V intake. Such strategies can have a public health impact on underserved populations.

A natively paired antibody library yields drug leads with higher sensitivity and specificity than a randomly paired antibody library.

PubMed

Adler, Adam S; Bedinger, Daniel; Adams, Matthew S; Asensio, Michael A; Edgar, Robert C; Leong, Renee; Leong, Jackson; Mizrahi, Rena A; Spindler, Matthew J; Bandi, Srinivasa Rao; Huang, Haichun; Tawde, Pallavi; Brams, Peter; Johnson, David S

2018-04-01

Deep sequencing and single-chain variable fragment (scFv) yeast display methods are becoming more popular for discovery of therapeutic antibody candidates in mouse B cell repertoires. In this study, we compare a deep sequencing and scFv display method that retains native heavy and light chain pairing with a related method that randomly pairs heavy and light chain. We performed the studies in a humanized mouse, using interleukin 21 receptor (IL-21R) as a test immunogen. We identified 44 high-affinity binder scFv with the native pairing method and 100 high-affinity binder scFv with the random pairing method. 30% of the natively paired scFv binders were also discovered with the randomly paired method, and 13% of the randomly paired binders were also discovered with the natively paired method. Additionally, 33% of the scFv binders discovered only in the randomly paired library were initially present in the natively paired pre-sort library. Thus, a significant proportion of "randomly paired" scFv were actually natively paired. We synthesized and produced 46 of the candidates as full-length antibodies and subjected them to a panel of binding assays to characterize their therapeutic potential. 87% of the antibodies were verified as binding IL-21R by at least one assay. We found that antibodies with native light chains were more likely to bind IL-21R than antibodies with non-native light chains, suggesting a higher false positive rate for antibodies from the randomly paired library. Additionally, the randomly paired method failed to identify nearly half of the true natively paired binders, suggesting a higher false negative rate. We conclude that natively paired libraries have critical advantages in sensitivity and specificity for antibody discovery programs.

Discounts on fruit and vegetables combined with a space management intervention increased sales in supermarkets.

PubMed

Toft, U; Winkler, L L; Mikkelsen, B E; Bloch, P; Glümer, C

2017-04-01

To examine the effects of two interventions on consumer purchases of fruits and vegetables (F&V) on the Danish island of Bornholm: a 20% discount on F&V combined with improved shelf-space allocation, and improved shelf-space allocation alone. A space management intervention to promote F&V sales was performed in two large discount supermarkets on Bornholm in Denmark for 3 months (September-November 2012). In addition, a 20% discount on F&V was introduced for 3 months in one of the supermarkets ('space + price'). The effect was evaluated using sales data from the two intervention supermarkets and three control supermarkets from the same supermarket chain but in Odsherred, Denmark (control area). Both the effect on sales of fresh F&V and potential unhealthy substitution effects were evaluated using multi-level regression analyses. During the price intervention period, the index number for sales of fresh vegetables increased by 22.2% (P=0.001) in the 'space + price' intervention supermarket compared with the control supermarkets. Furthermore, the index number for the sale of organic fresh fruit and vegetables increased by 12.1% (P=0.04) and the sale of the total amount of fruit and vegetables (fresh, frozen, dried and canned) increased by 15.3% (P=0.01) compared with the control supermarkets. In the 'space only' intervention supermarket no significant increase in the sale of fruit and vegetables was found. No unhealthy substitution effects were found. In conclusion, a 20% price reduction on F&V significantly increased sales of F&V. The effect was most pronounced on vegetables and no negative/unhealthy substitution effects were found.

A natively paired antibody library yields drug leads with higher sensitivity and specificity than a randomly paired antibody library

PubMed Central

Adler, Adam S.; Bedinger, Daniel; Adams, Matthew S.; Asensio, Michael A.; Edgar, Robert C.; Leong, Renee; Leong, Jackson; Mizrahi, Rena A.; Spindler, Matthew J.; Bandi, Srinivasa Rao; Huang, Haichun; Brams, Peter; Johnson, David S.

2018-01-01

ABSTRACT Deep sequencing and single-chain variable fragment (scFv) yeast display methods are becoming more popular for discovery of therapeutic antibody candidates in mouse B cell repertoires. In this study, we compare a deep sequencing and scFv display method that retains native heavy and light chain pairing with a related method that randomly pairs heavy and light chain. We performed the studies in a humanized mouse, using interleukin 21 receptor (IL-21R) as a test immunogen. We identified 44 high-affinity binder scFv with the native pairing method and 100 high-affinity binder scFv with the random pairing method. 30% of the natively paired scFv binders were also discovered with the randomly paired method, and 13% of the randomly paired binders were also discovered with the natively paired method. Additionally, 33% of the scFv binders discovered only in the randomly paired library were initially present in the natively paired pre-sort library. Thus, a significant proportion of “randomly paired” scFv were actually natively paired. We synthesized and produced 46 of the candidates as full-length antibodies and subjected them to a panel of binding assays to characterize their therapeutic potential. 87% of the antibodies were verified as binding IL-21R by at least one assay. We found that antibodies with native light chains were more likely to bind IL-21R than antibodies with non-native light chains, suggesting a higher false positive rate for antibodies from the randomly paired library. Additionally, the randomly paired method failed to identify nearly half of the true natively paired binders, suggesting a higher false negative rate. We conclude that natively paired libraries have critical advantages in sensitivity and specificity for antibody discovery programs. PMID:29376776

Iron-mediated changes in phytoplankton photosynthetic competence during SOIREE

NASA Astrophysics Data System (ADS)

Boyd, P. W.; Abraham, E. R.

Active fluorescence (fast repetition rate fluorometry, FRRF) was used to follow the photosynthetic response of the phytoplankton community during the 13-day Southern Ocean Iron RElease Experiment (SOIREE). This in situ iron enrichment was conducted in the polar waters of the Australasian-Pacific sector of the Southern Ocean in February 1999. Iron fertilisation of these high nitrate low chlorophyll (HNLC) waters resulted in an increase in the photosynthetic competence ( Fv/ Fm) of the resident cells from around 0.20 to greater than 0.60 (i.e. close to the theoretical maximum) by 10/11 days after the first enrichment. Although a significant iron-mediated response in Fv/ Fm was detected as early as 24 h after the initial fertilisation, the increase in Fv/ Fm to double ambient levels took 6 days. This response was five-fold slower than observed in iron enrichments (in situ and in vitro) in the HNLC waters of the subarctic and equatorial Pacific. Although little is known about the relationship between water temperature and Fv/ Fm, it is likely that low water temperatures — and possibly the deep mixed layer — were responsible for this slow response time. During SOIREE, the photosynthetic competence of the resident phytoplankton in iron-enriched waters increased at dissolved iron levels above 0.2 nM, suggesting that iron limitation was alleviated at this concentration. Increases in Fv/ Fm of cells within four algal size classes suggested that all taxa displayed a photosynthetic response to iron enrichment. Other physiological proxies of algal iron stress (such as flavodoxin levels in diatoms) exhibited different temporal trends to iron-enrichment than Fv/ Fm during the time-course of SOIREE. The relationship between Fv/ Fm, algal growth rate and such proxies in Southern Ocean waters is discussed.

Evaluating the Use of Antibody Variable Region (Fv) Charge as a Risk Assessment Tool for Predicting Typical Cynomolgus Monkey Pharmacokinetics*

PubMed Central

Bumbaca Yadav, Daniela; Sharma, Vikas K.; Boswell, Charles Andrew; Hotzel, Isidro; Tesar, Devin; Shang, Yonglei; Ying, Yong; Fischer, Saloumeh K.; Grogan, Jane L.; Chiang, Eugene Y.; Urban, Konnie; Ulufatu, Sheila; Khawli, Leslie A.; Prabhu, Saileta; Joseph, Sean; Kelley, Robert F.

2015-01-01

The pharmacokinetic (PK) behavior of monoclonal antibodies in cynomolgus monkeys (cynos) is generally translatable to that in humans. Unfortunately, about 39% of the antibodies evaluated for PKs in cynos have fast nonspecific (or non-target-mediated) clearance (in-house data). An empirical model relating variable region (Fv) charge and hydrophobicity to cyno nonspecific clearance was developed to gauge the risk an antibody would have for fast nonspecific clearance in the monkey. The purpose of this study was to evaluate the predictability of this empirical model on cyno nonspecific clearance with antibodies specifically engineered to have either high or low Fv charge. These amino acid changes were made in the Fv region of two test antibodies, humAb4D5-8 and anti-lymphotoxin α. The humAb4D5-8 has a typical nonspecific clearance in cynos, and by making it more positively charged, the antibody acquires fast nonspecific clearance, and making it less positively charged did not impact its clearance. Anti-lymphotoxin α has fast nonspecific clearance in cynos, and making it more positively charged caused it to clear even faster, whereas making it less positively charged caused it to clear slower and within the typical range. These trends in clearance were also observed in two other preclinical species, mice and rats. The effect of modifying Fv charge on subcutaneous bioavailability was also examined, and in general bioavailability was inversely related to the direction of the Fv charge change. Thus, modifying Fv charge appears to impact antibody PKs, and the changes tended to correlate with those predicted by the empirical model. PMID:26491012

Single chain variable fragment displaying M13 phage library functionalized magnetic microsphere-based protein equalizer for human serum protein analysis.

PubMed

Zhu, Guijie; Zhao, Peng; Deng, Nan; Tao, Dingyin; Sun, Liangliang; Liang, Zhen; Zhang, Lihua; Zhang, Yukui

2012-09-18

Single chain variable fragment (scFv) displaying the M13 phage library was covalently immobilized on magnetic microspheres and used as a protein equalizer for the treatment of human serum. First, scFv displaying M13 phage library functionalized magnetic microspheres (scFv@M13@MM) was incubated with a human serum sample. Second, captured proteins on scFv@M13@MM were eluted with 2 M NaCl, 50 mM glycine-hydrochloric acid (Gly-HCl), and 20% (v/v) acetonitrile with 0.5% (v/v) trifluoroacetic acid in sequence. Finally, the tightly bonded proteins were released by the treatment with thrombin. The eluates were first analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with silver staining. Results indicated that the difference of protein concentration was reduced obviously in NaCl and Gly-HCl fractions compared with untreated human serum sample. The eluates were also digested with trypsin, followed by online 2D-strong cation exchange (SCX)-RPLC-ESI-MS/MS analysis. Results demonstrated that the number of proteins identified from an scFv@M13@MM treated human serum sample was improved 100% compared with that from the untreated sample. In addition, the spectral count of 10 high abundance proteins (serum albumin, serotransferrin, α-2-macroglobulin, α-1-antitrypsin, apolipoprotein B-100, Ig γ-2 chain C region, haptoglobin, hemopexin, α-1-acid glycoprotein 1, and α-2-HS-glycoprotein) decreased evidently after scFv@M13@MM treatment. All these results demonstrate that scFv@M13@MM could efficiently remove high-abundance proteins, reduce the protein concentration difference of human serum, and result in more protein identification.

Socioeconomic indicators and frequency of traditional food, junk food, and fruit and vegetable consumption amongst Inuit adults in the Canadian Arctic.

PubMed

Hopping, B N; Erber, E; Mead, E; Sheehy, T; Roache, C; Sharma, S

2010-10-01

Increasing consumption of non-nutrient-dense foods (NNDF), decreasing consumption of traditional foods (TF) and low consumption of fruit and vegetables (FV) may contribute to increasing chronic disease rates amongst Inuit. The present study aimed to assess the daily frequency and socioeconomic and demographic factors influencing consumption of TF, FV and NNDF amongst Inuit adults in Nunavut, Canada. Using a cross-sectional study design and random household sampling in three communities in Nunavut, a food frequency questionnaire developed for the population was used to assess frequency of NNDF, TF and FV consumption amongst Inuit adults. Socioeconomic status (SES) was assessed by education level, ownership of items in working condition, and whether or not people in the household were employed or on income support. Mean frequencies of daily consumption were compared across gender and age groups, and associations with socioeconomic indicators were analysed using logistic regression. Two hundred and eleven participants (36 men, 175 women; mean (standard deviation) ages 42.1 (15.0) and 42.2 (13.2) years, respectively; response rate 69-93%) completed the study. Mean frequencies of consumption for NNDF, TF and FV were 6.3, 1.9 and 1.6 times per day, respectively. On average, participants ≤50 years consumed NNDF (P=0.003) and FV (P=0.01) more frequently and TF (P=0.01) less frequently than participants >50 years. Education was positively associated with FV consumption and negatively associated with TF consumption. Households on income support were more likely to consume TF and NNDF. These results support the hypothesis that the nutrition transition taking place amongst Inuit in Nunavut results in elevated consumption of NNDF compared with TF and FV. © 2010 The Authors. Journal compilation © 2010 The British Dietetic Association Ltd.

Enhancement and Analysis of Human Antiaflatoxin B1 (AFB1) scFv Antibody-Ligand Interaction Using Chain Shuffling.

PubMed

Rangnoi, Kuntalee; Choowongkomon, Kiattawee; O'Kennedy, Richard; Rüker, Florian; Yamabhai, Montarop

2018-06-06

A human antiaflatoxin B1 (AFB1) scFv antibody (yAFB1-c3), selected from a naı̈ve human phage-displayed scFv library, was used as a template for improving and analysis of antibody-ligand interactions using the chain-shuffling technique. The variable-heavy and variable-light (VH/VL)-shuffled library was constructed from the VH of 25 preselected clones recombined with the VL of yAFB1-c3 and vice versa. Affinity selection from these libraries demonstrated that the VH domain played an important role in the binding of scFv to free AFB1. Therefore, in the next step, VH-shuffled scFv library was constructed from variable-heavy (VH) chain repertoires, amplified from the naı̈ve library, recombined with the variable-light (VL) chain of the clone yAFB1-c3. This library was then used to select a specific scFv antibody against soluble AFB1 by a standard biopanning method. Three clones that showed improved binding properties were isolated. Amino acid sequence analysis indicated that the improved clones have amino acid mutations in framework 1 (FR1) and the complementarity determining region (CDR1) of the VH chain. One clone, designated sAFH-3e3, showed 7.5-fold improvement in sensitivity over the original scFv clone and was selected for molecular binding studies with AFB1. Homology modeling and molecular docking were used to compare the binding of this and the original clones. The results confirmed that VH is more important than VL for AFB1 binding.

Identification of inhibitory scFv antibodies targeting fibroblast activation protein utilizing phage display functional screens

PubMed Central

Zhang, Jiping; Valianou, Matthildi; Simmons, Heidi; Robinson, Matthew K.; Lee, Hyung-Ok; Mullins, Stefanie R.; Marasco, Wayne A.; Adams, Gregory P.; Weiner, Louis M.; Cheng, Jonathan D.

2013-01-01

Fibroblast activation protein (FAP) is a serine protease selectively expressed on tumor stromal fibroblasts in epithelial carcinomas and is important in cancer growth, adhesion, and metastases. As FAP enzymatic activity is a potent therapeutic target, we aimed to identify inhibitory antibodies. Using a competitive inhibition strategy, we used phage display techniques to identify 53 single-chain variable fragments (scFvs) after three rounds of panning against FAP. These scFvs were expressed and characterized for binding to FAP by surface plasmon resonance and flow cytometry. Functional assessment of these antibodies yielded an inhibitory scFv antibody, named E3, which could attenuate 35% of FAP cleavage of the fluorescent substrate Ala-Pro-7-amido-4-trifluoromethylcoumarin compared with nonfunctional scFv control. Furthermore, a mutant E3 scFv was identified by yeast affinity maturation. It had higher affinity (4-fold) and enhanced inhibitory effect on FAP enzyme activity (3-fold) than E3. The application of both inhibitory anti-FAP scFvs significantly affected the formation of 3-dimensional FAP-positive cell matrix, as demonstrated by reducing the fibronectin fiber orientation from 41.18% (negative antibody control) to 34.06% (E3) and 36.15% (mutant E3), respectively. Thus, we have identified and affinity-maturated the first scFv antibody capable of inhibiting FAP function. This scFv antibody has the potential to disrupt the role of FAP in tumor invasion and metastasis.—Zhang, J., Valianou, M., Simmons, H., Robinson, M. K., Lee, H.-O., Mullins, S. R., Marasco, W. A., Adams, G. P., Weiner, L. M., Cheng, J. D. Identification of inhibitory ScFv antibodies targeting fibroblast activation protein utilizing phage display functional screens. PMID:23104982

Differential Impact of LPG-and PG-Deficient Leishmania major Mutants on the Immune Response of Human Dendritic Cells

PubMed Central

Jayakumar, Asha; Hickerson, Suzanne; Mostrom, Janet; Turco, Salvatore J.; Beverley, Stephen M.; McDowell, Mary Ann

2015-01-01

Background Leishmania major infection induces robust interleukin-12 (IL12) production in human dendritic cells (hDC), ultimately resulting in Th1-mediated immunity and clinical resolution. The surface of Leishmania parasites is covered in a dense glycocalyx consisting of primarily lipophosphoglycan (LPG) and other phosphoglycan-containing molecules (PGs), making these glycoconjugates the likely pathogen-associated molecular patterns (PAMPS) responsible for IL12 induction. Methodology/Principal Findings Here we explored the role of parasite glycoconjugates on the hDC IL12 response by generating L. major Friedlin V1 mutants defective in LPG alone, (FV1 lpg1-), or generally deficient for all PGs, (FV1 lpg2-). Infection with metacyclic, infective stage, L. major or purified LPG induced high levels of IL12B subunit gene transcripts in hDCs, which was abrogated with FV1 lpg1- infections. In contrast, hDC infections with FV1 lpg2- displayed increased IL12B expression, suggesting other PG-related/LPG2 dependent molecules may act to dampen the immune response. Global transcriptional profiling comparing WT, FV1 lpg1-, FV1 lpg2- infections revealed that FV1 lpg1- mutants entered hDCs in a silent fashion as indicated by repression of gene expression. Transcription factor binding site analysis suggests that LPG recognition by hDCs induces IL-12 in a signaling cascade resulting in Nuclear Factor κ B (NFκB) and Interferon Regulatory Factor (IRF) mediated transcription. Conclusions/Significance These data suggest that L. major LPG is a major PAMP recognized by hDC to induce IL12-mediated protective immunity and that there is a complex interplay between PG-baring Leishmania surface glycoconjugates that result in modulation of host cellular IL12. PMID:26630499

Short-term effects of povidone iodine and sodium fluoride therapy on plaque levels and microbiome diversity.

PubMed

Reilly, C; Goettl, M; Steinmetz, M; Nikrad, J; Jones, R S

2016-03-01

The objective of this study is to evaluate the effect of short-term changes in the oral microbial ecology of dental plaque and plaque levels after topical treatment of a combination of 10% povidone iodine (PI) and 5% sodium fluoride varnish (FV). A single group design intervention study on 12 pediatric patients, who underwent two baseline plaques samplings before the intervention, were enrolled in the study. A modified mixed dentition Silness-Löe plaque index score was used to assess plaque accumulation and microbial composition was assessed by amplicon sequencing analysis of the 16S rRNA V4 region. Dental plaque accumulation (P = 0.0424) was reduced after 1 week using PI/FV application. This reduction was not observed between the two double-baseline visits. 16S rRNA analysis showed that the single PI/FV therapy did not have dramatic shifts in the plaque microbiome community depicted by hierarchical cluster and principle component analysis. More subtle changes were found when analyzing the Shannon diversity index after the application of PI/FV vs two baselines prior to combination therapy. The bacteria within the dental biofilms showed resilience in maintaining the overall community diversity but reduced biofilm accumulation following PI/FV therapy. Repeated uses of PI/FV may augment plaque control during dental rehabilitation in children. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Anthelmintic effects of the essential oil of fennel (Foeniculum vulgare Mill., Apiaceae) against Schistosoma mansoni.

PubMed

Wakabayashi, Kamila A L; de Melo, Nathalya I; Aguiar, Daniela P; de Oliveira, Pollyanna F; Groppo, Milton; da Silva Filho, Ademar A; Rodrigues, Vanderlei; Cunha, Wilson R; Tavares, Denise C; Magalhães, Lizandra G; Crotti, Antônio E M

2015-07-01

Foeniculum vulgare Mill. (Apiaceae), known as fennel, is a widespread aromatic herbaceous plant, and its essential oil is used as additive in the food, pharmaceutical, cosmetic, and perfume industries. The in vitro antischistosomal activity and cytotoxic effects against V79 cells of the essential oil of F. vulgare cultivated in southeastern Brazil (FV-EO) was investigated. The FV-EO was obtained by hydrodistillation and characterized by GC-FID and GC/MS analyses. (E)-Anethole (69.8%) and limonene (22.5%) were identified as the major constituents. Its anthelmintic activity against Schistosoma mansoni was evaluated at concentrations of 10, 50, and 100 μg/ml, and it was found to be active against adult S. mansoni worms, although it was less effective than the positive control praziquantel (PZQ) in terms of separation of the coupled pairs, mortality, and decreased motor activity. However, FV-EO elicited an interesting dose-dependent reduction in the number of S. mansoni eggs. On their own, (E)-anethole and the limonene enantiomers were much less effective than FV-EO and PZQ. An XTT-cytotoxicity-based assay evidenced no FV-EO cytotoxicity against V79 cells. In summary, FV-EO displayed moderate in vitro schistosomicidal activity against adult S. mansoni worms, exerted remarkable inhibitory effects on the egg development, and was of low toxicity. Copyright © 2015 Verlag Helvetica Chimica Acta AG, Zürich.

Lunch Salad Bars in New Orleans’ Middle and High Schools: Student Intake of Fruit and Vegetables

PubMed Central

Johnson, Carolyn C.; Myers, Leann; Mundorf, Adrienne R.; O’Malley, Keelia; Spruance, Lori Andersen; Harris, Diane M.

2017-01-01

The school lunch salad bar (SB) is a recommended food environmental strategy to increase access to, and consumption of fruit and vegetables (F/V). In a study to examine use of school lunch SBs, middle and high school students provided data via the Automated Self-Administered 24-h dietary recall (24HDR) tool for kids (ASA24-Kids-2012), a web-based data collection platform. Kilocalories were computed, food groups were assigned and F/V sources were obtained. Students (n = 718) from 12 schools with SBs and nine schools without SBs were approximately 87% African American, over 64% female and most were 7th and 8th graders. SB school students had higher median energy consumption at lunch but a higher percent of non-SB students reported eating fruit at lunch compared to SB students. Most students reporting eating F/V at lunch obtained F/V from the cafeteria main line; only 19.6% reported eating F/V exclusively from the SB. In SB schools median intake of cups F/V was higher among students using the SB (0.92) compared to those not using the SB (0.53). Results of this study are mixed, but encouraging. Additional factors, e.g., nutrition education, marketing, and kinds of foods offered on the SB need to be examined for potential influence on SB use. PMID:28406472

A chimeric antigen receptor for TRAIL-receptor 1 induces apoptosis in various types of tumor cells.

PubMed

Kobayashi, Eiji; Kishi, Hiroyuki; Ozawa, Tatsuhiko; Hamana, Hiroshi; Nakagawa, Hidetoshi; Jin, Aishun; Lin, Zhezhu; Muraguchi, Atsushi

2014-10-31

Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) and its associated receptors (TRAIL-R/TR) are attractive targets for cancer therapy because TRAIL induces apoptosis in tumor cells through TR while having little cytotoxicity on normal cells. Therefore, many agonistic monoclonal antibodies (mAbs) specific for TR have been produced, and these induce apoptosis in multiple tumor cell types. However, some TR-expressing tumor cells are resistant to TR-specific mAb-induced apoptosis. In this study, we constructed a chimeric antigen receptor (CAR) of a TRAIL-receptor 1 (TR1)-specific single chain variable fragment (scFv) antibody (TR1-scFv-CAR) and expressed it on a Jurkat T cell line, the KHYG-1 NK cell line, and human peripheral blood lymphocytes (PBLs). We found that the TR1-scFv-CAR-expressing Jurkat cells killed target cells via TR1-mediated apoptosis, whereas TR1-scFv-CAR-expressing KHYG-1 cells and PBLs killed target cells not only via TR1-mediated apoptosis but also via CAR signal-induced cytolysis, resulting in cytotoxicity on a broader range if target cells than with TR1-scFv-CAR-expressing Jurkat cells. The results suggest that TR1-scFv-CAR could be a new candidate for cancer gene therapy. Copyright © 2014 Elsevier Inc. All rights reserved.

Development of a Novel Human scFv Against EGFR L2 Domain by Phage Display Technology.

PubMed

Rahbarnia, Leila; Farajnia, Safar; Babaei, Hossein; Majidi, Jafar; Veisi, Kamal; Khosroshahi, Shiva Ahdi; Tanomand, Asghar

2017-01-01

Epidermal growth factor receptor (EGFR) as a transmembrane tyrosine kinase receptor frequently overexpresses in tumors with epithelial origin. The L2 domain from extracellular part of EGFR is involved in ligand binding and the blockage of this domain prevents activation of related signaling pathways. This study was aimed to develop a novel human scFv against EGFR L2 domain as a promising target for cancer therapy. The L2 recombinant protein was purified and used for panning a human scFv phage library (Tomlinson I). In this study, a novel screening strategy was applied to select clones with high binding and enrichment of rare specific phage clones of the L2 protein. After five biopanning rounds several specific clones were isolated which among them one phage clone with high binding was purified for further analysis. The specific interaction of selected clone against target antigen was confirmed by ELISA and western blotting. Immunofluorescence staining showed that purified scFv binds to A431 cells surface, displaying EGFR surface receptor. In the present study, we isolated for the first time a novel human scFv against EGFR L2 domain. This study can be the groundwork for developing more effective diagnostic and therapeutic agents against EGFR overexpressing cancers using this novel human anti-L2 ScFv. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

Consumption Frequency of Foods Away from Home Linked with Higher Body Mass Index and Lower Fruit and Vegetable Intake among Adults: A Cross-Sectional Study

PubMed Central

Seguin, Rebecca A.; Aggarwal, Anju; Vermeylen, Francoise; Drewnowski, Adam

2016-01-01

Introduction. Consumption of foods prepared away from home (FAFH) has grown steadily since the 1970s. We examined the relationship between FAFH and body mass index (BMI) and fruit and vegetable (FV) consumption. Methods. Frequency of FAFH, daily FV intake, height and weight, and sociodemographic data were collected using a telephone survey in 2008-2009. Participants included a representative sample of 2,001 adult men and women (mean age 54 ± 15 years) residing in King County, WA, with an analytical sample of 1,570. Frequency of FAFH was categorized as 0-1, 2–4, or 5+ times per week. BMI was calculated from self-reported height and weight. We examined the relationship between FAFH with FV consumption and BMI using multivariate models. Results. Higher frequency of FAFH was associated with higher BMI, after adjusting for age, income, education, race, smoking, marital status, and physical activity (women: p = 0.001; men: p = 0.003). There was a negative association between frequency of FAFH and FV consumption. FAFH frequency was significantly (p < 0.001) higher among males than females (43.1% versus 54.0% eating out 0-1 meal per week, resp.). Females reported eating significantly (p < 0.001) more FV than males. Conclusion. Among adults, higher frequency of FAFH was related to higher BMI and less FV consumption. PMID:26925111

Anti-Aβ single-chain variable fragment antibodies exert synergistic neuroprotective activities in Drosophila models of Alzheimer's disease

PubMed Central

Fernandez-Funez, Pedro; Zhang, Yan; Sanchez-Garcia, Jonatan; de Mena, Lorena; Khare, Swati; Golde, Todd E.; Levites, Yona; Rincon-Limas, Diego E.

2015-01-01

Both active and passive immunotherapy protocols decrease insoluble amyloid-ß42 (Aß42) peptide in animal models, suggesting potential therapeutic applications against the main pathological trigger in Alzheimer's disease (AD). However, recent clinical trials have reported no significant benefits from humanized anti-Aß42 antibodies. Engineered single-chain variable fragment antibodies (scFv) are much smaller and can easily penetrate the brain, but identifying the most effective scFvs in murine AD models is slow and costly. We show here that scFvs against the N- and C-terminus of Aß42 (scFv9 and scFV42.2, respectively) that decrease insoluble Aß42 in CRND mice are neuroprotective in Drosophila models of Aß42 and amyloid precursor protein neurotoxicity. Both scFv9 and scFv42.2 suppress eye toxicity, reduce cell death in brain neurons, protect the structural integrity of dendritic terminals in brain neurons and delay locomotor dysfunction. Additionally, we show for the first time that co-expression of both anti-Aß scFvs display synergistic neuroprotective activities, suggesting that combined therapies targeting distinct Aß42 epitopes can be more effective than targeting a single epitope. Overall, we demonstrate the feasibility of using Drosophila as a first step for characterizing neuroprotective anti-Aß scFvs in vivo and identifying scFv combinations with synergistic neuroprotective activities. PMID:26253732

Multi-channeled single chain variable fragment (scFv) based microfluidic device for explosives detection.

PubMed

Charles, Paul T; Davis, Jasmine; Adams, André A; Anderson, George P; Liu, Jinny L; Deschamps, Jeffrey R; Kusterbeck, Anne W

2015-11-01

The development of explosives detection technologies has increased significantly over the years as environmental and national security agencies implement tighter pollution control measures and methods for improving homeland security. 2, 4, 6-Trinitrotoluene (TNT), known primarily as a component in munitions, has been targeted for both its toxicity and carcinogenic properties that if present at high concentrations can be a detriment to both humans, marine and plant ecosystems. Enabling end users with environmental detection and monitoring systems capable of providing real-time, qualitative and quantitative chemical analysis of these toxic compounds would be extremely beneficial. Reported herein is the development of a multi-channeled microfluidic device immobilized with single chain fragment variable (scFv) recombinant proteins specific for the explosive, TNT. Fluorescence displacement immunoassays performed under constant flow demonstrated trace level sensitivity and specificity for TNT. The utility of three multi-channeled devices immobilized with either (1) scFv recombinant protein, (2) biotinylated-scFv (bt-scFv) and (3) monoclonal anti-TNT (whole IgG molecule) were investigated and compared. Fluorescence dose response curves, crossreactivity measurements and limits of detection (LOD) for TNT were determined. Fluorescence displacement immunoassays for TNT in natural seawater demonstrated detection limits at sub-parts-per-billion levels (0.5 ppb) utilizing the microfluidic device with immobilized bt-scFv. Published by Elsevier B.V.

Lunch Salad Bars in New Orleans' Middle and High Schools: Student Intake of Fruit and Vegetables.

PubMed

Johnson, Carolyn C; Myers, Leann; Mundorf, Adrienne R; O'Malley, Keelia; Spruance, Lori Andersen; Harris, Diane M

2017-04-13

The school lunch salad bar (SB) is a recommended food environmental strategy to increase access to, and consumption of fruit and vegetables (F/V). In a study to examine use of school lunch SBs, middle and high school students provided data via the Automated Self-Administered 24-h dietary recall (24HDR) tool for kids (ASA24-Kids-2012), a web-based data collection platform. Kilocalories were computed, food groups were assigned and F/V sources were obtained. Students (n = 718) from 12 schools with SBs and nine schools without SBs were approximately 87% African American, over 64% female and most were 7th and 8th graders. SB school students had higher median energy consumption at lunch but a higher percent of non-SB students reported eating fruit at lunch compared to SB students. Most students reporting eating F/V at lunch obtained F/V from the cafeteria main line; only 19.6% reported eating F/V exclusively from the SB. In SB schools median intake of cups F/V was higher among students using the SB (0.92) compared to those not using the SB (0.53). Results of this study are mixed, but encouraging. Additional factors, e.g., nutrition education, marketing, and kinds of foods offered on the SB need to be examined for potential influence on SB use.

Novel Immunocytokine IL12-SS1 (Fv) Inhibits Mesothelioma Tumor Growth in Nude Mice

PubMed Central

Kim, Heungnam; Gao, Wei; Ho, Mitchell

2013-01-01

Mesothelin is a glycosylphosphatidylinositol-anchored glycoprotein that is highly expressed on the cell surface of malignant mesothelioma. Monoclonal antibodies against mesothelin are being evaluated for the treatment of mesothelioma. Immunocytokines represent a novel class of armed antibodies. To provide an alternative approach to current mesothelin-targeted antibody therapies, we have developed a novel immunocytokine based on interleukin-12 (IL12) and the SS1 Fv specific for mesothelin. IL12 possesses potent anti-tumor activity in a wide variety of solid tumors. The newly-developed recombinant immunocytokine, IL12-SS1 (Fv), was produced in insect cells using a baculovirus-insect cell expression system. The SS1 single-chain Fv was fused to the C terminus of the p35 subunit of IL12 through a short linker (GSADGG). The single-chain IL12-SS1 (Fv) immunocytokine bound native mesothelin proteins on malignant mesothelioma (NCI-H226) and ovarian (OVCAR-3) cells as well as recombinant mesothelin on A431/H9 cells. The immunocytokine retained sufficient bioactivity of IL12 and significantly inhibited human malignant mesothelioma (NCI-H226) grown in the peritoneal cavity of nude mice and showed comparable anti-tumor activity to that of the SS1P immunotoxin. IL12-SS1 (Fv) is the first reported immunocytokine to mesothelin-positive tumors and may be an attractive addition to mesothelin-targeted cancer therapies. PMID:24260587

Quantitative and nutritional characterization of fruit and vegetable waste from marketplace: a potential use as bovine feedstuff?

PubMed

Angulo, Joaquin; Mahecha, Liliana; Yepes, Sergio A; Yepes, Angela M; Bustamante, Gilberto; Jaramillo, Harold; Valencia, Edward; Villamil, Tomás; Gallo, Jorge

2012-03-01

There are different sources for the generation of solid waste, and marketplaces are considered one of them. Fruit and vegetable waste (FV) from a marketplace in Colombia was quantitatively and nutritionally characterized to contribute to its use in bovine feeding and to contribute minimizing its environmental impact. The evaluation was carried out 7 days per week during 4 periods of the year. FV was grouped by cluster analysis using SAS(®) 2006. FV was composed of 43% fruit, 30% vegetables and 27% stems, leaves, leaf wrappers, corncobs, roots, refuse and others. FV was defined in four main groups. On average, FV contained 10% crude protein (CP), 36.6% neutral detergent fiber (NDF), 29.6% acid detergent fiber (ADF), 87.8% ruminal degradability at 24 h, 3657 kcal/kg, 0.59% calcium (Ca(+2)), and 0.21% phosphorous (P). There were no statistical differences between days or between periods of evaluation (p > 0.05) for CP or for Ca(+2). As for NDF and ADF, there were statistically significant differences between periods but not between days. The microbiological parameters only increased when the humidity was up to 12%. FV represents a potential feedstuff for bovine feeding, and its recycling could avoid the discharge of a large amount of waste to landfills, which would minimize its environmental impact. Copyright © 2010. Published by Elsevier Ltd.

Bioinformatics identification and transcript profile analysis of the mitogen-activated protein kinase gene family in the diploid woodland strawberry Fragaria vesca

PubMed Central

Wei, Wei; Chai, Zhuangzhuang; Xie, Yinge; Gao, Kuan; Cui, Mengyuan; Jiang, Ying

2017-01-01

Mitogen-activated protein kinases (MAPKs) play essential roles in mediating biotic and abiotic stress responses in plants. However, the MAPK gene family in strawberry has not been systematically characterized. Here, we performed a genome-wide survey and identified 12 MAPK genes in the Fragaria vesca genome. Protein domain analysis indicated that all FvMAPKs have typical protein kinase domains. Sequence alignments and phylogenetic analysis classified the FvMAPK genes into four different groups. Conserved motif and exon-intron organization supported the evolutionary relationships inferred from the phylogenetic analysis. Analysis of the stress-related cis-regulatory element in the promoters and subcellular localization predictions of FvMAPKs were also performed. Gene transcript profile analysis showed that the majority of the FvMAPK genes were ubiquitously transcribed in strawberry leaves after Podosphaera aphanis inoculation and after treatment with cold, heat, drought, salt and the exogenous hormones abscisic acid, ethephon, methyl jasmonate, and salicylic acid. RT-qPCR showed that six selected FvMAPK genes comprehensively responded to various stimuli. Additionally, interaction networks revealed that the crucial signaling transduction controlled by FvMAPKs may be involved in the biotic and abiotic stress responses. Our results may provide useful information for future research on the function of the MAPK gene family and the genetic improvement of strawberry resistance to environmental stresses. PMID:28562633

Improved Soluble ScFv ELISA Screening Approach for Antibody Discovery Using Phage Display Technology.

PubMed

Tohidkia, Mohammad R; Sepehri, Maryam; Khajeh, Shirin; Barar, Jaleh; Omidi, Yadollah

2017-09-01

Phage display technology (PDT) is a powerful tool for the isolation of recombinant antibody (Ab) fragments. Using PDT, target molecule-specific phage-Ab clones are enriched through the "biopanning" process. The individual specific binders are screened by the monoclonal scFv enzyme-linked immunosorbent assay (ELISA) that may associate with inevitable false-negative results. Thus, in this study, three strategies were investigated for optimization of the scFvs screening using Tomlinson I and J libraries, including (1) optimizing the expression of functional scFvs, (2) improving the sensitivity of ELISA, and (3) preparing different samples containing scFvs. The expression of all scFv Abs was significantly enhanced when scFv clones were cultivated in the terrific broth (TB) medium at the optimum temperature of 30 °C. The protein A-conjugated with horseradish peroxidase (HRP) was found to be a well-suited reagent for the detection of Ag-bound scFvs in comparison with either anti-c-myc Ab or the mixing procedure. Based on our findings, it seems there is no universal media supplement for an improved expression of all scFvs derived from both Tomlinson I and J libraries. We thus propose that expression of scFv fragments in a microplate scale is largely dependent on a variety of parameters, in particular the scFv clones and relevant sequences.

Expression, production, and renaturation of a functional single-chain variable antibody fragment (scFv) against human ICAM-1

PubMed Central

Sun, H.; Wu, G.M.; Chen, Y.Y.; Tian, Y.; Yue, Y.H.; Zhang, G.L.

2014-01-01

Intercellular adhesion molecule-1 (ICAM-1) is an important factor in the progression of inflammatory responses in vivo. To develop a new anti-inflammatory drug to block the biological activity of ICAM-1, we produced a monoclonal antibody (Ka=4.19×10−8 M) against human ICAM-1. The anti-ICAM-1 single-chain variable antibody fragment (scFv) was expressed at a high level as inclusion bodies in Escherichia coli. We refolded the scFv (Ka=2.35×10−7 M) by ion-exchange chromatography, dialysis, and dilution. The results showed that column chromatography refolding by high-performance Q Sepharose had remarkable advantages over conventional dilution and dialysis methods. Furthermore, the anti-ICAM-1 scFv yield of about 60 mg/L was higher with this method. The purity of the final product was greater than 90%, as shown by denaturing gel electrophoresis. Enzyme-linked immunosorbent assay, cell culture, and animal experiments were used to assess the immunological properties and biological activities of the renatured scFv. PMID:24919171

Efficient recovery of the functional IP10-scFv fusion protein from inclusion bodies with an on-column refolding system.

PubMed

Guo, Jun-Qing; Li, Qing-Mei; Zhou, Ji-Yong; Zhang, Gai-Ping; Yang, Yan-Yan; Xing, Guang-Xu; Zhao, Dong; You, Shang-You; Zhang, Chu-Yu

2006-01-01

A functional IP10-scFv fusion protein retaining the antibody specificity for acidic isoferritin and chemokine function was produced at high level in Esherichia coli (E. coli). IP10-scFv gene from the recombinant plasmid pc3IP104c9 was subcloned into pET28a fused to N-terminal His-tag sequence in frame and overexpressed in E. coli BL21(DE3). With an on-column refolding procedure based on Ni-chelating chromatography, the active fusion protein was recovered efficiently from inclusion bodies with a refolding yield of approximate 45% confirmed by spectrophotometer. The activity of refolded IP10-scFv was determined through sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western blotting and enzyme-linked immunosorbent assay. The results showed the fusion protein retains the specific binding activity to AIF with an affinity constant of 4.48x10(-8) M as well as the chemokine function of IP-10. The overall yield of IP10-scFv with bioactivity in E. coli flask culture was more than 40 mg/L.

FvSet2 regulates fungal growth, pathogenicity, and secondary metabolism in Fusarium verticillioides.

PubMed

Gu, Qin; Wang, Zhenzhong; Sun, Xiao; Ji, Tiantian; Huang, Hai; Yang, Yang; Zhang, Hao; Tahir, Hafiz Abdul Samad; Wu, Liming; Wu, Huijun; Gao, Xuewen

2017-10-01

Histone H3 lysine 36 methylation (H3K36me) is generally associated with activation of gene expression in most eukaryotic cells. However, the function of H3K36me in filamentous fungi is largely unknown. Set2 is the sole lysine histone methyltransferase (KHMTase) enzyme responsible for the methylation of H3K36 in Saccharomyces cerevisiae. In the current study, we identified a single ortholog of S. cerevisiae Set2 in Fusarium verticillioides. We report that FvSet2 is responsible for the trimethylation of H3K36 (H3K36me3). The FvSET2 deletion mutant (ΔFvSet2) showed significant defects in vegetative growth, FB 1 biosynthesis, pigmentation, and fungal virulence. Furthermore, trimethylation of H3K36 was found to be important for active transcription of genes involved in FB 1 and bikaverin biosyntheses. These data indicate that FvSet2 plays an important role in the regulation of secondary metabolism, vegetative growth and fungal virulence in F. verticillioides. Copyright © 2017 Elsevier Inc. All rights reserved.

ESCRT-mediated Uptake and Degradation of Brain-targeted α-synuclein Single Chain Antibody Attenuates Neuronal Degeneration In Vivo

PubMed Central

Spencer, Brian; Emadi, Sharareh; Desplats, Paula; Eleuteri, Simona; Michael, Sarah; Kosberg, Kori; Shen, Jay; Rockenstein, Edward; Patrick, Christina; Adame, Anthony; Gonzalez, Tania; Sierks, Michael; Masliah, Eliezer

2014-01-01

Parkinson's disease and dementia with Lewy bodies are neurodegenerative disorders characterized by accumulation of α-synuclein (α-syn). Recently, single-chain fragment variables (scFVs) have been developed against individual conformational species of α-syn. Unlike more traditional monoclonal antibodies, these scFVs will not activate or be endocytosed by Fc receptors. For this study, we investigated an scFV directed against oligomeric α-syn fused to the LDL receptor-binding domain from apolipoprotein B (apoB). The modified scFV showed enhanced brain penetration and was imported into neuronal cells through the endosomal sorting complex required for transport (ESCRT) pathway, leading to lysosomal degradation of α-syn aggregates. Further analysis showed that the scFV was effective at ameliorating neurodegenerative pathology and behavioral deficits observed in the mouse model of dementia with Lewy bodies/Parkinson's disease. Thus, the apoB modification had the effect of both increasing accumulation of the scFV in the brain and directing scFV/α-syn complexes for degradation through the ESCRT pathway, leading to improved therapeutic potential of immunotherapy. PMID:25008355

ESCRT-mediated uptake and degradation of brain-targeted α-synuclein single chain antibody attenuates neuronal degeneration in vivo.

PubMed

Spencer, Brian; Emadi, Sharareh; Desplats, Paula; Eleuteri, Simona; Michael, Sarah; Kosberg, Kori; Shen, Jay; Rockenstein, Edward; Patrick, Christina; Adame, Anthony; Gonzalez, Tania; Sierks, Michael; Masliah, Eliezer

2014-10-01

Parkinson's disease and dementia with Lewy bodies are neurodegenerative disorders characterized by accumulation of α-synuclein (α-syn). Recently, single-chain fragment variables (scFVs) have been developed against individual conformational species of α-syn. Unlike more traditional monoclonal antibodies, these scFVs will not activate or be endocytosed by Fc receptors. For this study, we investigated an scFV directed against oligomeric α-syn fused to the LDL receptor-binding domain from apolipoprotein B (apoB). The modified scFV showed enhanced brain penetration and was imported into neuronal cells through the endosomal sorting complex required for transport (ESCRT) pathway, leading to lysosomal degradation of α-syn aggregates. Further analysis showed that the scFV was effective at ameliorating neurodegenerative pathology and behavioral deficits observed in the mouse model of dementia with Lewy bodies/Parkinson's disease. Thus, the apoB modification had the effect of both increasing accumulation of the scFV in the brain and directing scFV/α-syn complexes for degradation through the ESCRT pathway, leading to improved therapeutic potential of immunotherapy.

Tumor-targeting CTL expressing a single-chain Fv specific for VEGFR2.

PubMed

Kanagawa, Naoko; Yanagawa, Tatsuya; Mukai, Yohei; Yoshioka, Yasuo; Okada, Naoki; Nakagawa, Shinsaku

2010-03-26

Cytotoxic T lymphocytes (CTL) are critical effector cells in tumor immunity. Adoptive transfer therapy with in vitro-expanded tumor-specific CTL is a promising approach for preventing cancer metastasis and recurrence. Transferred CTL are not effective in clinical trials, however, due to inadequate tumor-infiltration. Therefore, the development of functionally modified CTL, such as tumor-targeting CTL, is widely desired. Here, we designed the tumor-targeting CTL expressing a single-chain antibody fragment (scFv-CTL) specific for vascular endothelial growth factor receptor 2 (VEGFR2/flk1) by transducing the CTL with a retroviral vector. The scFv-CTL bound to VEGFR2/flk1-expressing cells and retained their cytotoxic activity against tumor cells. In addition, adoptive transfer of scFv-CTL into tumor-bearing mice effectively suppressed tumor growth due to the augmented accumulation of the transferred CTL in the tumor tissue. These findings indicate that the creation of CTL capable of targeting tumor vascular endothelial cells by scFv-expression technique is considerably promising for improvement of efficacy in adoptive immunotherapy. Copyright (c) 2010 Elsevier Inc. All rights reserved.

Search for maximal flavor violating scalars in same-charge lepton pairs in pp collisions at sqrt[s]=1.96 TeV.

PubMed

Aaltonen, T; Adelman, J; Akimoto, T; Albrow, M G; Alvarez González, B; Amerio, S; Amidei, D; Anastassov, A; Annovi, A; Antos, J; Aoki, M; Apollinari, G; Apresyan, A; Arisawa, T; Artikov, A; Ashmanskas, W; Attal, A; Aurisano, A; Azfar, F; Azzi-Bacchetta, P; Azzurri, P; Bacchetta, N; Badgett, W; Barbaro-Galtieri, A; Barnes, V E; Barnett, B A; Baroiant, S; Bar-Shalom, S; Bartsch, V; Bauer, G; Beauchemin, P-H; Bedeschi, F; Bednar, P; Behari, S; Bellettini, G; Bellinger, J; Belloni, A; Benjamin, D; Beretvas, A; Beringer, J; Berry, T; Bhatti, A; Binkley, M; Bisello, D; Bizjak, I; Blair, R E; Blocker, C; Blumenfeld, B; Bocci, A; Bodek, A; Boisvert, V; Bolla, G; Bolshov, A; Bortoletto, D; Boudreau, J; Boveia, A; Brau, B; Bridgeman, A; Brigliadori, L; Bromberg, C; Brubaker, E; Budagov, J; Budd, H S; Budd, S; Burkett, K; Busetto, G; Bussey, P; Buzatu, A; Byrum, K L; Cabrera, S; Campanelli, M; Campbell, M; Canelli, F; Canepa, A; Carlsmith, D; Carosi, R; Carrillo, S; Carron, S; Casal, B; Casarsa, M; Castro, A; Catastini, P; Cauz, D; Cavalli-Sforza, M; Cerri, A; Cerrito, L; Chang, S H; Chen, Y C; Chertok, M; Chiarelli, G; Chlachidze, G; Chlebana, F; Cho, K; Chokheli, D; Chou, J P; Choudalakis, G; Chuang, S H; Chung, K; Chung, W H; Chung, Y S; Ciobanu, C I; Ciocci, M A; Clark, A; Clark, D; Compostella, G; Convery, M E; Conway, J; Cooper, B; Copic, K; Cordelli, M; Cortiana, G; Crescioli, F; Cuenca Almenar, C; Cuevas, J; Culbertson, R; Cully, J C; Dagenhart, D; Datta, M; Davies, T; de Barbaro, P; De Cecco, S; Deisher, A; De Lentdecker, G; De Lorenzo, G; Dell'orso, M; Demortier, L; Deng, J; Deninno, M; De Pedis, D; Derwent, P F; Di Giovanni, G P; Dionisi, C; Di Ruzza, B; Dittmann, J R; D'Onofrio, M; Donati, S; Dong, P; Donini, J; Dorigo, T; Dube, S; Efron, J; Erbacher, R; Errede, D; Errede, S; Eusebi, R; Fang, H C; Farrington, S; Fedorko, W T; Feild, R G; Feindt, M; Fernandez, J P; Ferrazza, C; Field, R; Flanagan, G; Forrest, R; Forrester, S; Franklin, M; Freeman, J C; Furic, I; Gallinaro, M; Galyardt, J; Garberson, F; Garcia, J E; Garfinkel, A F; Genser, K; Gerberich, H; Gerdes, D; Giagu, S; Giakoumopolou, V; Giannetti, P; Gibson, K; Gimmell, J L; Ginsburg, C M; Giokaris, N; Giordani, M; Giromini, P; Giunta, M; Glagolev, V; Glenzinski, D; Gold, M; Goldschmidt, N; Golossanov, A; Gomez, G; Gomez-Ceballos, G; Goncharov, M; González, O; Gorelov, I; Goshaw, A T; Goulianos, K; Gresele, A; Grinstein, S; Grosso-Pilcher, C; Grundler, U; Guimaraes da Costa, J; Gunay-Unalan, Z; Haber, C; Hahn, K; Hahn, S R; Halkiadakis, E; Hamilton, A; Han, B-Y; Han, J Y; Handler, R; Happacher, F; Hara, K; Hare, D; Hare, M; Harper, S; Harr, R F; Harris, R M; Hartz, M; Hatakeyama, K; Hauser, J; Hays, C; Heck, M; Heijboer, A; Heinemann, B; Heinrich, J; Henderson, C; Herndon, M; Heuser, J; Hewamanage, S; Hidas, D; Hill, C S; Hirschbuehl, D; Hocker, A; Hou, S; Houlden, M; Hsu, S-C; Huffman, B T; Hughes, R E; Husemann, U; Huston, J; Incandela, J; Introzzi, G; Iori, M; Ivanov, A; Iyutin, B; James, E; Jayatilaka, B; Jeans, D; Jeon, E J; Jindariani, S; Johnson, W; Jones, M; Joo, K K; Jun, S Y; Jung, J E; Junk, T R; Kamon, T; Kar, D; Karchin, P E; Kato, Y; Kephart, R; Kerzel, U; Khotilovich, V; Kilminster, B; Kim, D H; Kim, H S; Kim, J E; Kim, M J; Kim, S B; Kim, S H; Kim, Y K; Kimura, N; Kirsch, L; Klimenko, S; Klute, M; Knuteson, B; Ko, B R; Koay, S A; Kondo, K; Kong, D J; Konigsberg, J; Korytov, A; Kotwal, A V; Kraus, J; Kreps, M; Kroll, J; Krumnack, N; Kruse, M; Krutelyov, V; Kubo, T; Kuhlmann, S E; Kuhr, T; Kulkarni, N P; Kusakabe, Y; Kwang, S; Laasanen, A T; Lai, S; Lami, S; Lammel, S; Lancaster, M; Lander, R L; Lannon, K; Lath, A; Latino, G; Lazzizzera, I; Lecompte, T; Lee, J; Lee, J; Lee, Y J; Lee, S W; Lefèvre, R; Leonardo, N; Leone, S; Levy, S; Lewis, J D; Lin, C; Lin, C S; Linacre, J; Lindgren, M; Lipeles, E; Lister, A; Litvintsev, D O; Liu, T; Lockyer, N S; Loginov, A; Loreti, M; Lovas, L; Lu, R-S; Lucchesi, D; Lueck, J; Luci, C; Lujan, P; Lukens, P; Lungu, G; Lyons, L; Lys, J; Lysak, R; Lytken, E; Mack, P; Macqueen, D; Madrak, R; Maeshima, K; Makhoul, K; Maki, T; Maksimovic, P; Malde, S; Malik, S; Manca, G; Manousakis, A; Margaroli, F; Marino, C; Marino, C P; Martin, A; Martin, M; Martin, V; Martínez, M; Martínez-Ballarín, R; Maruyama, T; Mastrandrea, P; Masubuchi, T; Mattson, M E; Mazzanti, P; McFarland, K S; McIntyre, P; McNulty, R; Mehta, A; Mehtala, P; Menzemer, S; Menzione, A; Merkel, P; Mesropian, C; Messina, A; Miao, T; Miladinovic, N; Miles, J; Miller, R; Mills, C; Milnik, M; Mitra, A; Mitselmakher, G; Miyake, H; Moed, S; Moggi, N; Moon, C S; Moore, R; Morello, M; Movilla Fernandez, P; Mülmenstädt, J; Mukherjee, A; Muller, Th; Mumford, R; Murat, P; Mussini, M; Nachtman, J; Nagai, Y; Nagano, A; Naganoma, J; Nakamura, K; Nakano, I; Napier, A; Necula, V; Neu, C; Neubauer, M S; Nielsen, J; Nodulman, L; Norman, M; Norniella, O; Nurse, E; Oh, S H; Oh, Y D; Oksuzian, I; Okusawa, T; Oldeman, R; Orava, R; Osterberg, K; Pagan Griso, S; Pagliarone, C; Palencia, E; Papadimitriou, V; Papaikonomou, A; Paramonov, A A; Parks, B; Pashapour, S; Patrick, J; Pauletta, G; Paulini, M; Paus, C; Pellett, D E; Penzo, A; Phillips, T J; Piacentino, G; Piedra, J; Pinera, L; Pitts, K; Plager, C; Pondrom, L; Portell, X; Poukhov, O; Pounder, N; Prakoshyn, F; Pronko, A; Proudfoot, J; Ptohos, F; Punzi, G; Pursley, J; Rademacker, J; Rahaman, A; Rajaraman, A; Ramakrishnan, V; Ranjan, N; Redondo, I; Reisert, B; Rekovic, V; Renton, P; Rescigno, M; Richter, S; Rimondi, F; Ristori, L; Robson, A; Rodrigo, T; Rogers, E; Rolli, S; Roser, R; Rossi, M; Rossin, R; Roy, P; Ruiz, A; Russ, J; Rusu, V; Saarikko, H; Safonov, A; Sakumoto, W K; Salamanna, G; Saltó, O; Santi, L; Sarkar, S; Sartori, L; Sato, K; Savoy-Navarro, A; Scheidle, T; Schlabach, P; Schmidt, E E; Schmidt, M A; Schmidt, M P; Schmitt, M; Schwarz, T; Scodellaro, L; Scott, A L; Scribano, A; Scuri, F; Sedov, A; Seidel, S; Seiya, Y; Semenov, A; Sexton-Kennedy, L; Sfyria, A; Shalhout, S Z; Shapiro, M D; Shears, T; Shepard, P F; Sherman, D; Shimojima, M; Shochet, M; Shon, Y; Shreyber, I; Sidoti, A; Sinervo, P; Sisakyan, A; Slaughter, A J; Slaunwhite, J; Sliwa, K; Smith, J R; Snider, F D; Snihur, R; Soderberg, M; Soha, A; Somalwar, S; Sorin, V; Spalding, J; Spinella, F; Spreitzer, T; Squillacioti, P; Stanitzki, M; St Denis, R; Stelzer, B; Stelzer-Chilton, O; Stentz, D; Strologas, J; Stuart, D; Suh, J S; Sukhanov, A; Sun, H; Suslov, I; Suzuki, T; Taffard, A; Takashima, R; Takeuchi, Y; Tanaka, R; Tecchio, M; Teng, P K; Terashi, K; Thom, J; Thompson, A S; Thompson, G A; Thomson, E; Tipton, P; Tiwari, V; Tkaczyk, S; Toback, D; Tokar, S; Tollefson, K; Tomura, T; Tonelli, D; Torre, S; Torretta, D; Tourneur, S; Trischuk, W; Tu, Y; Turini, N; Ukegawa, F; Uozumi, S; Vallecorsa, S; van Remortel, N; Varganov, A; Vataga, E; Vázquez, F; Velev, G; Vellidis, C; Veszpremi, V; Vidal, M; Vidal, R; Vila, I; Vilar, R; Vine, T; Vogel, M; Volobouev, I; Volpi, G; Würthwein, F; Wagner, P; Wagner, R G; Wagner, R L; Wagner-Kuhr, J; Wagner, W; Wakisaka, T; Wallny, R; Wang, S M; Warburton, A; Waters, D; Weinberger, M; Wester, W C; Whitehouse, B; Whiteson, D; Wicklund, A B; Wicklund, E; Williams, G; Williams, H H; Wilson, P; Winer, B L; Wittich, P; Wolbers, S; Wolfe, C; Wright, T; Wu, X; Wynne, S M; Yagil, A; Yamamoto, K; Yamaoka, J; Yamashita, T; Yang, C; Yang, U K; Yang, Y C; Yao, W M; Yeh, G P; Yoh, J; Yorita, K; Yoshida, T; Yu, G B; Yu, F; Yu, I; Yu, S S; Yun, J C; Zanello, L; Zanetti, A; Zaw, I; Zhang, X; Zheng, Y; Zucchelli, S

2009-01-30

Models of maximal flavor violation (MxFV) in elementary particle physics may contain at least one new scalar SU(2) doublet field Phi(FV)=(eta(0),eta(+)) that couples the first and third generation quarks (q_(1), q_(3)) via a Lagrangian term L(FV)=xi(13)Phi(FV)q(1)q(3). These models have a distinctive signature of same-charge top-quark pairs and evade flavor-changing limits from meson mixing measurements. Data corresponding to 2 fb(-1) collected by the Collider Dectector at Fermilab II detector in pp[over ] collisions at sqrt[s]=1.96 TeV are analyzed for evidence of the MxFV signature. For a neutral scalar eta(0) with m_(eta;(0))=200 GeV/c(2) and coupling xi(13)=1, approximately 11 signal events are expected over a background of 2.1+/-1.8 events. Three events are observed in the data, consistent with background expectations, and limits are set on the coupling xi(13) for m(eta(0)=180-300 GeV/c(2).

Prevalence of 1691G>A FV mutation in females from Bosnia and Herzegovina - a preliminary report

PubMed Central

Yaljevac, Amina; Mehić, Bakir; Kiseljaković, Emina; Ibrulj, Slavka; Garstka, Agnieszka; Adler, Grazyna

2013-01-01

Factor V is the liver-synthesized multidomain glycoprotein encoded by a gene localised on chromosome 1q23. The point mutation 1691G>A in this gene results in formation of an altered protein of V Factor resistant to activated protein C (APC) cleavage. This mutation alone is the most frequent cause of inborn thrombophilia and the most widely acknowledged genetic risk factor for venous thrombosis in a Caucasian population. This study was designed to provide the first estimate of the frequency of the allele 1691A FV in the Bosnian female population. The 1691G>A FV mutation was examined by polymerase chain reaction-restriction fragment length polymorphism, in a group of 67 women, mean age of 58.6 years with no history of cardiovascural incident. Our findings revealed an absence of the mutated allele 1691A FV in the studied group. This is the first report on the 1691G>A FV mutation in a population from Bosnia and Herzegovina. Further research is needed to establish prevalence of the mutated allele in the population from Bosnia and Herzegovina. PMID:23448608

Construction of a scFv Library with Synthetic, Non-combinatorial CDR Diversity.

PubMed

Bai, Xuelian; Shim, Hyunbo

2017-01-01

Many large synthetic antibody libraries have been designed, constructed, and successfully generated high-quality antibodies suitable for various demanding applications. While synthetic antibody libraries have many advantages such as optimized framework sequences and a broader sequence landscape than natural antibodies, their sequence diversities typically are generated by random combinatorial synthetic processes which cause the incorporation of many undesired CDR sequences. Here, we describe the construction of a synthetic scFv library using oligonucleotide mixtures that contain predefined, non-combinatorially synthesized CDR sequences. Each CDR is first inserted to a master scFv framework sequence and the resulting single-CDR libraries are subjected to a round of proofread panning. The proofread CDR sequences are assembled to produce the final scFv library with six diversified CDRs.

Friendly fire: redirecting herpes simplex virus-1 for therapeutic applications.

PubMed

Advani, S J; Weichselbaum, R R; Whitley, R J; Roizman, B

2002-09-01

Herpes simplex virus-1 (HSV-1) is a relatively large double-stranded DNA virus encoding at least 89 proteins with well characterized disease pathology. An understanding of the functions of viral proteins together with the ability to genetically engineer specific viral mutants has led to the development of attenuated HSV-1 for gene therapy. This review highlights the progress in creating attenuated genetically engineered HSV-1 mutants that are either replication competent (viral non-essential gene deleted) or replication defective (viral essential gene deleted). The choice between a replication-competent or -defective virus is based on the end-goal of the therapeutic intervention. Replication-competent HSV-1 mutants have primarily been employed as antitumor oncolytic viruses, with the lytic nature of the virus harnessed to destroy tumor cells selectively. In replacement gene therapy, replication-defective viruses have been utilized as delivery vectors. The advantages of HSV-1 vectors are that they infect quiescent and dividing cells efficiently and can encode for relatively large transgenes.

Operator-Friendly Technique and Quality Control Considerations for Indigo Colorimetric Measurement of Ozone Residual

EPA Science Inventory

Drinking water ozone disinfection systems measure ozone residual concentration, C, for regulatory compliance reporting of concentration-times-time (CT), and the resultant log-inactivation of virus, Giardia and Cryptosporidium. The indigotrisulfonate (ITS) colorimetric procedure i...

Sexual communication between early adolescents and their dating partners, parents, and best friends.

PubMed

Widman, Laura; Choukas-Bradley, Sophia; Helms, Sarah W; Golin, Carol E; Prinstein, Mitchell J

2014-01-01

This study assessed early adolescents' sexual communication with dating partners, parents, and best friends about six sexual health topics: condoms, birth control, sexually transmitted diseases (STDs), human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS), pregnancy, and abstinence/waiting. Using a school-based sample of 603 youth (ages 12 to 15; 57% female; 46% Caucasian), we examined communication differences across demographic and developmental factors, tested whether communication with parents and best friends was associated with greater communication with partners, and examined associations between communication and condom use. More than half of participants had not discussed any sexual topics with their dating partners (54%), and many had not communicated with parents (29%) or best friends (25%). On average, communication was more frequent among adolescents who were female, African American, older, and sexually active, despite some variation in subgroups across partner, parent, and friend communication. Importantly, communication with parents and friends--and the interaction between parent and friend communication--was associated with increased communication with dating partners. Further, among sexually active youth, increased sexual communication with partners was associated with more frequent condom use. Results highlight the importance of understanding the broader family and peer context surrounding adolescent sexual decision making and suggest a possible need to tailor sexual communication interventions.

‘Gay boy talk’ meets ‘girl talk’: HIV risk assessment assumptions in young gay men's sexual health communication with best friends

PubMed Central

Mutchler, Matt G.; McDavitt, Bryce

2011-01-01

Young adults, particularly young gay men (YGM), are vulnerable to human immunodeficiency virus (HIV). Yet, little is known about how YGM discuss sexual health issues with their friends (‘gay boy talk’). We conducted semi-structured interviews with YGM and their best friends (11 YGM/YGM dyads and 13 YGM/heterosexual female dyads). In this paper, we examine risk assessment assumptions conveyed within YGM's communication about sexual health with their friends and how, if at all, the sexual scripts guiding these assumptions may differ between YGM and young women. Findings demonstrated that, while these young adults clearly intended to support their friends and promote safer sex, they also conveyed assumptions about HIV risk assessment, especially regarding sexual partner selection, that may actually increase their friends’ risk for HIV infection. Since inaccurate HIV risk assessment assumptions were transmitted via sexual health communication between peers, it is suggested that such assumptions may need to be addressed in HIV prevention programs working with YGM and their friends. Further, gender differences were identified within the sexual scripts shared between YGM and their friends, suggesting that such interventions should be tailored to the specific needs of different friendship networks. PMID:21059803

Ludwik Gross, Sarah Stewart, and the 1950s discoveries of Gross murine leukemia virus and polyoma virus.

PubMed

Morgan, Gregory J

2014-12-01

The Polish-American scientist Ludwik Gross made two important discoveries in the early 1950s. He showed that two viruses - murine leukemia virus and parotid tumor virus - could cause cancer when they were injected into susceptible animals. At first, Gross's discoveries were greeted with skepticism: it seemed implausible that viruses could cause a disease as complex as cancer. Inspired by Gross's initial experiments, similar results were obtained by Sarah Stewart and Bernice Eddy who later renamed the parotid tumor virus SE polyoma virus after finding it could cause many different types of tumors in mice, hamsters, and rats. Eventually the "SE" was dropped and virologists adopted the name "polyoma virus." After Gross's work was published, additional viruses capable of causing solid tumors or blood-borne tumors in mice were described by Arnold Graffi, Charlotte Friend, John Moloney and others. By 1961, sufficient data had been accumulated for Gross to confidently publish an extensive monograph--Oncogenic Viruses--the first history of tumor virology, which became a standard reference work and marked the emergence of tumor virology as a distinct, legitimate field of study. Copyright © 2014 Elsevier Ltd. All rights reserved.

Evaluation of eco-friendly zwitterionic detergents for enveloped virus inactivation.

PubMed

Conley, Lynn; Tao, Yinying; Henry, Alexis; Koepf, Edward; Cecchini, Douglas; Pieracci, John; Ghose, Sanchayita

2017-04-01

Inclusion of a detergent in protein biotherapeutic purification processes is a simple and very robust method for inactivating enveloped viruses. The detergent Triton X-100 has been used for many years and is part of the production process of several commercial therapeutic proteins. However, recent ecological studies have suggested that Triton X-100 and its break-down products can potentially behave as endocrine disrupters in aquatic organisms, raising concerns from an environmental impact perspective. As such, discharge of Triton X-100 into the waste water treatment plants is regulated in some jurisdictions, and alternative detergents for viral inactivation are required. In this work, we report on the identification and evaluation of more eco-friendly detergents as viable replacements for Triton X-100. Five detergent candidates with low to moderate environmental impact were initially identified and evaluated with respect to protein stability, followed by proof-of-concept virus inactivation studies using a model enveloped virus. From the set of candidates lauryldimethylamine N-oxide (LDAO) was identified as the most promising detergent due to its low ecotoxicity, robust anti-viral activity (LRV >4 at validation set-point conditions with X-MuLX), and absence of any negative impact on protein function. This detergent exhibited effective and robust virus inactivation in a broad range of protein concentrations, solution conductivities, pHs, and in several different cell culture fluid matrices. The only process parameter which correlated with reduced virus inactivation potency was LDAO concentration, and then only when the concentration was reduced to below the detergent's critical micelle concentration (CMC). Additionally, this work also demonstrated that LDAO was cleared to below detectable levels after Protein A affinity chromatography, making it suitable for use in a platform process that utilizes this chromatographic mode for protein capture. All these findings suggest that LDAO may be a practical alternative to Triton X-100 for use in protein therapeutic production processes for inactivating enveloped viruses. Biotechnol. Bioeng. 2017;114: 813-820. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

The relationship between sleep duration and fruit/vegetable intakes in UK adults: a cross-sectional study from the National Diet and Nutrition Survey.

PubMed

Noorwali, Essra A; Cade, Janet E; Burley, Victoria J; Hardie, Laura J

2018-04-27

There is increasing evidence to suggest an association between sleep and diet. The aim of the present study was to examine the association between sleep duration and fruit/vegetable (FV) intakes and their associated biomarkers in UK adults. Cross-sectional. Data from The National Diet and Nutrition Survey. 1612 adults aged 19-65 years were included, pregnant/breastfeeding women were excluded from the analyses. Sleep duration was assessed by self-report, and diet was assessed by 4-day food diaries, disaggregation of foods containing FV into their components was conducted to determine total FV intakes. Sleep duration was divided into: short (<7 hours/day), reference (7-8 hours/day) and long (>8 hours/day) sleep periods. Multiple regression adjusting for confounders was used for analyses where sleep duration was the exposure and FV intakes and their associated biomarkers were the outcomes. Restricted cubic spline models were developed to explore potential non-linear associations. In adjusted models, long sleepers (LS) consumed on average 28 (95% CI -50 to -6, p=0.01) g/day less of total FV compared to reference sleepers (RS), whereas short sleepers (SS) consumed 24 g/day less (95% CI -42 to -6, p=0.006) and had lower levels of FV biomarkers (total carotenoids, β-carotene and lycopene) compared to RS. Restricted cubic spline models showed that the association between sleep duration and FV intakes was non-linear (p<0.001) with RS having the highest intakes compared to SS and LS. The associations between sleep duration and plasma total carotenoids (p=0.0035), plasma vitamin C (p=0.009) and lycopene (p<0.001) were non-linear with RS having the highest levels. These findings show a link between sleep duration and FV consumption. This may have important implications for lifestyle and behavioural change policy. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

Determinants of fruit and vegetable consumption among children and adolescents: a review of the literature. Part II: qualitative studies.

PubMed

Krølner, Rikke; Rasmussen, Mette; Brug, Johannes; Klepp, Knut-Inge; Wind, Marianne; Due, Pernille

2011-10-14

Large proportions of children do not fulfil the World Health Organization recommendation of eating at least 400 grams of fruit and vegetables (FV) per day. To promote an increased FV intake among children it is important to identify factors which influence their consumption. Both qualitative and quantitative studies are needed. Earlier reviews have analysed evidence from quantitative studies. The aim of this paper is to present a systematic review of qualitative studies of determinants of children's FV intake. Relevant studies were identified by searching Anthropology Plus, Cinahl, CSA illumine, Embase, International Bibliography of the Social Sciences, Medline, PsycINFO, and Web of Science using combinations of synonyms for FV intake, children/adolescents and qualitative methods as search terms. The literature search was completed by December 1st 2010. Papers were included if they applied qualitative methods to investigate 6-18-year-olds' perceptions of factors influencing their FV consumption. Quantitative studies, review studies, studies reported in other languages than English, and non-peer reviewed or unpublished manuscripts were excluded. The papers were reviewed systematically using standardised templates for summary of papers, quality assessment, and synthesis of findings across papers. The review included 31 studies, mostly based on US populations and focus group discussions. The synthesis identified the following potential determinants for FV intake which supplement the quantitative knowledge base: Time costs; lack of taste guarantee; satiety value; appropriate time/occasions/settings for eating FV; sensory and physical aspects; variety, visibility, methods of preparation; access to unhealthy food; the symbolic value of food for image, gender identity and social interaction with peers; short term outcome expectancies. The review highlights numerous potential determinants which have not been investigated thoroughly in quantitative studies. Future large scale quantitative studies should attempt to quantify the importance of these factors. Further, mechanisms behind gender, age and socioeconomic differences in FV consumption are proposed which should be tested quantitatively in order to better tailor interventions to vulnerable groups. Finally, the review provides input to the conceptualisation and measurements of concepts (i.e. peer influence, availability in schools) which may refine survey instruments and theoretical frameworks concerning eating behaviours.

Older adults' perceptions of using iPads for improving fruit and vegetable intake: an exploratory study.

PubMed

Watkins, Ivan; Xie, Bo

2015-01-01

Fruit and vegetable (FV) consumption can improve older adults' health outcomes, but conventional interventions can be resource demanding and make it difficult to provide just-in-time intervention content. iPad-based interventions may help overcome these limitations, but little is known about how older adults might perceive and use iPads for FV consumption. To address this gap in the literature, we conducted a qualitative study to explore older adults' perceptions and use of iPads for improving FV consumption between February and August of 2012. Five focus group sessions each lasting 120 min were conducted with 22 older adult participants. During each session, participants received guided exposure and instruction on iPad use and then explored three iPad applications targeting FV consumption (MyFood, FiveADay Lite, and Whole Foods Market Recipes). Detailed notes from focus group interviews were analyzed with a grounded theory approach that applied a constant comparative method to enable themes to emerge from the data. Three themes were identified from the data regarding participants' baseline perceptions of iPads. These included (a) limited knowledge on iPad's functions, (b) iPads were intended for younger users, and (c) iPads were too expensive. Themes identified regarding participants' perceptions of iPads after guided exposure included (a) the touchscreen was easier to use than a computer mouse, (b) tapping the interface required practice, (c) portability was an asset in conjunction with functionality, (d) portability and functionality supported personal interests, (e) the difficulty of learning an iPad's functions varied, and (f) practice and instruction helped overcome fear of the iPad. Finally, participants recommended iPad app features that could help them overcome barriers to their FV intake. These included (a) locating inexpensive FV from nearby sources, (b) providing tailored food and recipe suggestions, and (c) tracking and communicating FV intake with a doctor. These findings have important implications for future research on mobile app-based eHealth interventions to improve older adults' FV intake.

Enhancement of antitumor activity of gammaretrovirus carrying IL-12 gene through genetic modification of envelope targeting HER2 receptor: a promising strategy for bladder cancer therapy.

PubMed

Tsai, Y-S; Shiau, A-L; Chen, Y-F; Tsai, H-T; Tzai, T-S; Wu, C-L

2010-01-01

The objective of this study was to develop an HER2-targeted, envelope-modified Moloney murine leukemia virus (MoMLV)-based gammaretroviral vector carrying interleukin (IL)-12 gene for bladder cancer therapy. It displayed a chimeric envelope protein containing a single-chain variable fragment (scFv) antibody to the HER2 receptor and carried the mouse IL-12 gene. The fragment of anti-erbB2scFv was constructed into the proline-rich region of the viral envelope of the packaging vector lacking a transmembrane subunit of the carboxyl terminal region of surface subunit. As compared with envelope-unmodified gammaretroviruses, envelope-modified ones had extended viral tropism to human HER2-expressing bladder cancer cell lines, induced apoptosis, and affected cell cycle progression despite lower viral titers. Moreover, animal studies showed that envelope-modified gammaretroviruses carrying IL-12 gene exerted higher antitumor activity in terms of retarding tumor growth and prolonging the survival of tumor-bearing mice than unmodified ones, which were associated with enhanced tumor cell apoptosis as well as increased intratumoral levels of IL-12, interferon-gamma, IL-1beta, and tumor necrosis factor-alpha proteins. Therefore, the antitumor activity of gammaretroviruses carrying the IL-12 gene was enhanced through genetic modification of the envelope targeting HER2 receptor, which may be a promising strategy for bladder cancer therapy.

Immune evasion strategies of ranaviruses and innate immune responses to these emerging pathogens.

PubMed

Grayfer, Leon; Andino, Francisco De Jesús; Chen, Guangchun; Chinchar, Gregory V; Robert, Jacques

2012-07-01

Ranaviruses (RV, Iridoviridae) are large double-stranded DNA viruses that infect fish, amphibians and reptiles. For ecological and commercial reasons, considerable attention has been drawn to the increasing prevalence of ranaviral infections of wild populations and in aquacultural settings. Importantly, RVs appear to be capable of crossing species barriers of numerous poikilotherms, suggesting that these pathogens possess a broad host range and potent immune evasion mechanisms. Indeed, while some of the 95-100 predicted ranavirus genes encode putative evasion proteins (e.g., vIFα, vCARD), roughly two-thirds of them do not share significant sequence identity with known viral or eukaryotic genes. Accordingly, the investigation of ranaviral virulence and immune evasion strategies is promising for elucidating potential antiviral targets. In this regard, recombination-based technologies are being employed to knock out gene candidates in the best-characterized RV member, Frog Virus (FV3). Concurrently, by using animal infection models with extensively characterized immune systems, such as the African clawed frog, Xenopus laevis, it is becoming evident that components of innate immunity are at the forefront of virus-host interactions. For example, cells of the macrophage lineage represent important combatants of RV infections while themselves serving as targets for viral infection, maintenance and possibly dissemination. This review focuses on the recent advances in the understanding of the RV immune evasion strategies with emphasis on the roles of the innate immune system in ranaviral infections.

USER FRIENDLY MODELS FOR EVALUATING HYDROGEOLOGIC BARRIERS TO VIRUSES

EPA Science Inventory

Impending regulations in U.S. EPA's forthcoming Ground Water Rule (Federal Register, 2000) will require public water systems (PWS) to more closely monitor their groundwater systems for contamination by pathogens. As part of this process, State resource managers will assess the se...

Fluorescent immunolabeling of cancer cells by quantum dots and antibody scFv fragment.

PubMed

Zdobnova, Tatiana A; Dorofeev, Sergey G; Tananaev, Piter N; Vasiliev, Roman B; Balandin, Taras G; Edelweiss, Eveline F; Stremovskiy, Oleg A; Balalaeva, Irina V; Turchin, Ilya V; Lebedenko, Ekaterina N; Zlomanov, Vladimir P; Deyev, Sergey M

2009-01-01

Semiconductor quantum dots (QDs) coupled with cancer-specific targeting ligands are new promising agents for fluorescent visualization of cancer cells. Human epidermal growth factor receptor 2/neu (HER2/neu), overexpressed on the surface of many cancer cells, is an important target for cancer diagnostics. Antibody scFv fragments as a targeting agent for direct delivery of fluorophores offer significant advantages over full-size antibodies due to their small size, lower cross-reactivity, and immunogenicity. We have used quantum dots linked to anti-HER2/neu 4D5 scFv antibody to label HER2/neu-overexpressing live cells. Labeling of target cells was shown to have high brightness, photostability, and specificity. The results indicate that construction based on quantum dots and scFv antibody can be successfully used for cancer cell visualization.

The role of the local retail food environment in fruit, vegetable and sugar-sweetened beverage consumption in Brazil.

PubMed

Duran, Ana Clara; de Almeida, Samuel Luna; Latorre, Maria do Rosario D O; Jaime, Patricia Constante

2016-04-01

To examine the relationship between the local retail food environment and consumption of fruits and vegetables (FV) and sugar-sweetened beverages (SSB) in São Paulo, Brazil, as well as the moderation effects of income in the studied relationships. Cross-sectional study design that drew upon neighbourhood- and individual-level data. For each participant, community (density and proximity) and community food environment (availability, variety, quality and price) measures of FV and SSB were assessed in retail food stores and specialized fresh produce markets within 1·6 km of their homes. Poisson generalized estimating equations (GEE) were used to model the associations of food consumption with food environment measures, adjusted by individual-level characteristics. São Paulo, Brazil. Adults (n 1842) residing in the same census tracts (n 52) in São Paulo, Brazil as those where the neighbourhood-level measures were taken. FV availability in neighbourhoods was associated with regular FV consumption (≥5 times/week; prevalence ratio=1·41; 95 % CI 1·19, 1·67). Regular FV consumption prevalence was significantly lower among lower-income individuals living in neighbourhoods with fewer supermarkets and fresh produce markets (P-interaction <0·05). A greater variety of SSB was associated with a 15 % increase in regular SSB consumption (≥5 times/week) prevalence, after adjustment for confounding variables. Our findings suggest that the local retail food environment is associated with FV and SSB consumption in a Brazilian urban sample.

Some notes on the Roman domination number and Italian domination number in graphs

NASA Astrophysics Data System (ADS)

Hajibaba, Maryam; Jafari Rad, Nader

2017-09-01

An Italian dominating function (or simply, IDF) on a graph G = (V, E) is a function f : V → {0, 1, 2} that satisfies the property that for every vertex v ∈ V, with f(v) = 0, Σ u∈N(v) f(u) ≥ 2. The weight of an Italian dominating function f is defined as w(f) = f(V ) = Σ u∈V f(u). The minimum weight among all of the Italian dominating functions on a graph G is called the Italian domination number of G, and is denoted by γI (G). A double Roman dominating function (or simply, DRDF) is a function f : V → {0, 1, 2, 3} having the property that if f(v) = 0 for a vertex v, then v has at least two adjacent vertices assigned 2 under f or one adjacent vertex assigned 3 under f, and if f(v) = 1, then v has at least one neighbor with f(w) ≥ 2. The weight of a DRDF f is defined as the sum f(V) = Σ v∈V f(v), and the minimum weight of a DRDF on G is the double Roman domination number of G, denoted by γdR (G). In this paper we show that γdR (G)/2 ≤ γI (G) ≤ 2γdR (G)/3, and characterize all trees T with γI (T) = 2γdR (T)/3.

Gardening for Health: Patterns of Gardening and Fruit and Vegetable Consumption Among the Navajo.

PubMed

Ornelas, India J; Osterbauer, Katie; Woo, Lisa; Bishop, Sonia K; Deschenie, Desiree; Beresford, Shirley A A; Lombard, Kevin

2018-05-19

American Indians, including Navajo, are disproportionately affected by obesity and diabetes, in part due to diet-related health behaviors. The purpose of this study was to assess the patterns of gardening and fruit and vegetable (FV) consumption among residents in two communities on the Navajo Nation in order to inform a community gardening intervention. We analyzed survey data collected from participants in the Yéego Gardening study conducted in two communities in the Navajo Nation (N = 169). We found that 51% of the sample gardened, and on average participants gardened 8.9 times per month. Lack of time (53%) and financial barriers, such as gas for transportation or irrigation (51 and 49%, respectively), were reported as barriers to gardening. Most participants reported low levels of self-efficacy (80%) and behavioral capability (82%) related to gardening. Those with higher levels of gardening self-efficacy and behavioral capability reported more frequent gardening. Average daily FV consumption was 2.5 servings. Most participants reported high levels of self-efficacy to eat FV daily (64%) and high behavioral capability to prepare FV (66%). There was a positive association between FV consumption and gardening, with those gardening more than 4 times per month eating about 1 more serving of FV per day than those gardening 4 or fewer times per month. Further research is needed to better understand how gardening can increase fruit and vegetable availability and consumption among residents of the Navajo Nation.

Perceived barriers towards healthy eating and their association with fruit and vegetable consumption.

PubMed

Mc Morrow, L; Ludbrook, A; Macdiarmid, J I; Olajide, D

2017-06-01

Improving dietary intakes is a key public health target. Perceived barriers to healthy eating (PBHE) are an important component of the Health Belief Model which aims to understand why individuals do not adopt preventive health measures. This study investigates the relationship between PBHE and reported fruit and vegetable (F&V) consumption. Data from the Scottish Health Survey 2008-11 (n = 8319) for PBHE and self-reported F&V consumption were used in Probit regression models to test the association between meeting the 400 g per day F&V recommendation and PBHE. Regression models show women who reported a lack of cooking skills were 10.4% less likely to meet the F&V recommendations (P = 0.001). Not liking the taste of healthy foods or finding them too boring (10.2%, P = 0.022), preparation time (5.6%, P = 0.020) or willpower (3.0%, P = 0.021) were also significant. For men, reporting not liking the taste of healthy foods or finding them too boring (6.8%, P = 0.02) was the only significant result. Price, a commonly reported PBHE, was not significantly associated with F&V consumption. Not all commonly reported perceived barriers to healthy eating are significantly associated with meeting the recommended F&V intake. © The Author 2016. Published by Oxford University Press on behalf of Faculty of Public Health. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Validating Stages of Change for Obesogenic Behaviors Across Filipino and Other Asian-American and Pacific Islander Adolescents.

PubMed

Fleary, Sasha A; Tagorda, M; Kim, S; Rathke, M; Nigg, C R

2018-06-01

Filipino adolescents are underrepresented in obesity research, although Filipinos are the second largest Asian-American and Pacific Islander (AAPI) subpopulation in the USA. An understanding of how well the theories of behavior change apply to Filipino and other AAPI adolescents is critical to addressing obesogenic behaviors in these groups. This study aimed to validate the transtheoretical model of behavior change (TTM) for physical activity (PA) and fruit and vegetable intake (FV) among a majority Filipino adolescent population. Adolescents in grades 9-11 (N = 159, 82.4% female) completed measures of PA and FV behaviors and PA and FV stages of change. One-way ANOVAs and Tukey's HSD post hoc tests were computed to assess the validity of the PA and FV stages of change with the respective behaviors. There was a significant effect for fruit (action > contemplation, preparation) and vegetable (maintenance, action > contemplation) intakes across the FV stages of change. There was a significant effect of strenuous PA (precontemplation/contemplation, preparation < action < maintenance) and moderate PA (precontemplation/contemplation < action, maintenance) across the PA stages of change. Some variability in associations emerged when the sample was stratified by gender. This study provides validity evidence for the TTM stages of change for FV and PA among Filipino and other AAPI adolescents. This validation, in turn, extends the generalizability of the stages of change construct to include this ethnic group and replicates other adolescent studies.

Detection of Metallothionein in Javanese Medaka (Oryzias javanicus), Using a scFv-Immobilized Protein Chip

PubMed Central

Lee, Euiyeon; Jeon, Hyunjin; Kang, Chungwon; Woo, Seonock; Yum, Seungshic; Kwon, Youngeun

2018-01-01

Environmental pollution by various industrial chemicals and biological agents poses serious risks to human health. Especially, marine contamination by potentially toxic elements (PTEs) has become a global concern in recent years. Many efforts have been undertaken to monitor the PTE contamination of the aquatic environment. However, there are few approaches available to assess the PTE exposure of aquatic organisms. In this research, we developed a strategy to evaluate the heavy metal exposure of marine organisms, by measuring the expression levels of metallothionein protein derived from Oryzias javanicus (OjaMT). OjaMT is a biomarker of heavy metal exposure because the expression level increases upon heavy metal exposure. The developed assay is based on a real-time, label-free surface plasmon resonance (SPR) measurement. Anti-OjaMT antibody and anti-OjaMT single-chain fragment of variable region (scFv) were used as detection probes. Two types of SPR sensor chips were fabricated, by immobilizing antibody or Cys3-tagged scFv (scFv-Cys3) in a controlled orientation and were tested for in situ label-free OjaMT detection. Compared to the antibody-presenting sensor chips, the scFv-presenting sensor chips showed improved performance, displaying enhanced sensitivity and enabling semi-quantitative detection. The portable SPR system combined with scFv-immobilized sensor chips is expected to provide an excellent point-of-care testing system that can monitor target biomarkers in real time. PMID:29614840

Microbiological characterization of vegetables and their rhizosphere soil in Eastern Poland.

PubMed

Kłapeć, Teresa; Wójcik-Fatla, Angelina; Cholewa, Alicja; Cholewa, Grażyna; Dutkiewicz, Jacek

2016-12-23

The aim of this study was to investigate the bacteriological quality of 5 kinds of vegetables (lettuce, dill, radish, beetroot, carrot) and their rhizosphere soil, originating from conventional farms located in the Lublin Province of Eastern Poland. A total number of 35 samples of fresh vegetables (FV) taken immediately from soil, 35 samples of soil from rhizosphere of these vegetables (SR) and 35 samples of vegetables sold at retail in the markets in Lublin (VR) were examined. The samples were analysed for the content of: aerobic mesophilic bacteria (AMB) grown at 30°C and 37°C, Gram-negative bacteria of Enterobacteriaceae family, faecal coliform (FC) bacteria, Salmonella spp., and Clostridium perfringens. Median AMB values determined at 30°C for FV, SR and VR were 5.27, 5.00, and 5.00 log 10 CFU g -1 , respectively, being significantly greater compared to those recorded at 37°C. The exceeding of the threshold value of 6.0 log 10 CFU g -1 proposed by Gelosa (1998) was noted only in 5 FV samples grown at 30°C (14.3%), and in 3 FV samples grown at 37°C (8.6%). The threshold value was never exceeded in SR and VR samples. Median concentrations of Enterobacteriaceae determined for FV, SR and VR were 4.03, 3.87, and 3.04 log 10 CFU g -1 , respectively. Eleven species of Enterobacteriaceae were identified in the FV, SR and VR samples. The percent of samples containing Escherichia coli was greatest for VR (22.9%), smaller for FV (17.1%) and smallest for SR (5.7%). The median concentrations of the faecal coliform bacteria (FC), determined by culture at 44°C, were low, amounting to 1.000 log 10 CFU g -1 for FV and SR and 0.00 for VR. All examined vegetable and soil samples tested negative for the presence of Salmonella. The median concentrations of Clostridium perfringens were low, amounting to 0.00 log 10 CFU g -1 for all categories of samples. This bacterium was relatively common in soil samples with the prevalence of 40.0%, but very rare in vegetable samples (occurring in 5.7% of FV and in none of VR samples). In conclusion, the results of the present study generally indicate that the microbiological quality of Polish vegetables grown on conventional farms is satisfactory and safe for consumers.

Poliomyelitis in MuLV-infected ICR-SCID mice after injection of basement membrane matrix contaminated with lactate dehydrogenase-elevating virus.

PubMed

Carlson Scholz, Jodi A; Garg, Rohit; Compton, Susan R; Allore, Heather G; Zeiss, Caroline J; Uchio, Edward M

2011-10-01

The arterivirus lactate dehydrogenase-elevating virus (LDV) causes life-long viremia in mice. Although LDV infection generally does not cause disease, infected mice that are homozygous for the Fv1(n) allele are prone to develop poliomyelitis when immunosuppressed, a condition known as age-dependent poliomyelitis. The development of age-dependent poliomyelitis requires coinfection with endogenous murine leukemia virus. Even though LDV is a common contaminant of transplantable tumors, clinical signs of poliomyelitis after inadvertent exposure to LDV have not been described in recent literature. In addition, LDV-induced poliomyelitis has not been reported in SCID or ICR mice. Here we describe the occurrence of poliomyelitis in ICR-SCID mice resulting from injection of LDV-contaminated basement membrane matrix. After exposure to LDV, a subset of mice presented with clinical signs including paresis, which was associated with atrophy of the hindlimb musculature, and tachypnea; in addition, some mice died suddenly with or without premonitory signs. Mice presenting within the first 6 mo after infection had regions of spongiosis, neuronal necrosis and astrocytosis of the ventral spinal cord, and less commonly, brainstem. Axonal degeneration of ventral roots prevailed in more chronically infected mice. LDV was identified by RT-PCR in 12 of 15 mice with typical neuropathology; positive antiLDV immunolabeling was identified in all PCR-positive animals (n = 7) tested. Three of 8 mice with neuropathology but no clinical signs were LDV negative by RT-PCR. RT-PCR yielded murine leukemia virus in spinal cords of all mice tested, regardless of clinical presentation or neuropathology.

Impact of Commercials on Food Preferences of Low-Income Minority Preschoolers

PubMed Central

Nicklas, Theresa A.; Goh, Eugenia Tsuei; Goodell, Lora S.; Acuff, Daniel S.; Reiher, Robert; Buday, Richard; Ottenbacher, Allison

2010-01-01

Objective To determine if fruit and vegetable (FV) commercials have an impact on preschool children’s preferences for specific FV. Design A year of extensive formative assessment was conducted to develop two 30-second commercials; “Judy Fruity” promoted apples and bananas and “Reggie Veggie” promoted broccoli and carrots. The commercials were embedded into a 15-minute TV program. FV preferences were assessed before and after four exposures to each of the commercials. Setting/Participants One hundred eighty-three preschool children (39% African-American; 61% Hispanic-American) from four Head Start centers in Houston, Texas. Main Outcome/Analysis A general linear model was used to assess whether FV preferences were significantly higher in the treatment group than the control group, controlling for baseline FV preferences, age, race and intervention dose in the model. Results There was a significantly higher preference for broccoli and carrots (p = 0.02) in the intervention group compared to the control group after multiple exposures to the vegetable commercial. Conclusions/Implications Data suggest that commercials promoting vegetables may be an effective strategy to influence young children’s preferences for vegetables. This may not be the case with fruit preferences which are already high in this age group. PMID:20851053

The Built Food Environment and Dietary Intake among African-American Adults

PubMed Central

Reitzel, Lorraine R.; Okamoto, Hiroe; Hernandez, Daphne C.; Regan, Seann D.; McNeill, Lorna H.; Obasi, Ezemenari M.

2016-01-01

Objectives The built food environment surrounding people's homes may influence their dietary intake. This exploratory study examined how the density of different sources of food in the residential environment was associated with dietary consumption among 77 African-American adults in Houston, Texas. Methods The number of fast-food-type restaurants, large grocery stores, and convenience-type stores within 2- and 5-mile residential buffers were divided by the respective areas to obtain food environment density variables. Intake of fruit and vegetables [FV], fiber [FI], and percent energy from fat [PEF] was assessed using National Health Interview Survey items. Covariate-adjusted regressions were used to assess relations of interest. Results Greater density of fast-food-type restaurants within 2 miles was associated with greater FV, FI, and PEF (ps ≤ .012); and for FV and FI within 5 miles (ps < .004). Density of large grocery stores was unrelated to intake. Greater density of convenience-type stores within 2 miles was negatively associated with FV and FI (ps ≤ .03); results became marginal at 5 miles for FV (p = .10) but not FI (p = .03). Conclusion Maximizing healthy offerings in venue-rich metropolitan areas might provide direction for policies to reduce obesity. PMID:26685808

Focus groups inform a web-based program to increase fruit and vegetable intake.

PubMed

Rolnick, Sharon J; Calvi, Josephine; Heimendinger, Jerianne; McClure, Jennifer B; Kelley, Mary; Johnson, Christine; Alexander, Gwen L

2009-11-01

To use focus groups to inform a web-based educational intervention for increased fruit and vegetable (FV) consumption. Twelve groups (participants=137, aged 21-65) were recruited from four geographically diverse health systems. Four groups were stratified by gender and eight by race (white and African American) and gender. Questions included perceptions of healthy eating, factors that encourage or serve as barriers to FV consumption and features preferred for a web-based educational intervention. Though knowledgeable about healthy eating, participants did not know how to achieve or always care about healthy nutritional choices. Motivators for FV consumption included being role models and health concerns. Barriers included: lack of time, expense and FV availability. Website preferences included: visuals, links, tailored materials, menu suggestions, goal setting assistance, printable summaries and built in motivation. The developers incorporated nearly all suggestions. Focus groups provided needs-based tactical strategies for an online, education intervention targeting factors to improve FV consumption. Focus groups can provide valuable input to inform interventions. Further, web-based programs' abilities to offer information without time or geographic constraints, with capacity for tailoring and tracking progress makes them a valuable addition in the arsenal of efforts to promote healthy behaviors.

Generation of anti-idiotype scFv for pharmacokinetic measurement in lymphoma patients treated with chimera anti-CD22 antibody SM03.

PubMed

Zhao, Qi; Wong, Pui-Fan; Lee, Susanna S T; Leung, Shui-On; Cheung, Wing-Tai; Wang, Jun-Zhi

2014-01-01

Pre-clinical and clinical studies of therapeutic antibodies require highly specific reagents to examine their immune responses, bio-distributions, immunogenicity, and pharmacodynamics in patients. Selective antigen-mimicking anti-idiotype antibody facilitates the assessment of therapeutic antibody in the detection, quantitation and characterization of antibody immune responses. Using mouse specific degenerate primer pairs and splenocytic RNA, we generated an idiotype antibody-immunized phage-displayed scFv library in which an anti-idiotype antibody against the therapeutic chimera anti-CD22 antibody SM03 was isolated. The anti-idiotype scFv recognized the idiotype of anti-CD22 antibody and inhibited binding of SM03 to CD22 on Raji cell surface. The anti-idiotype scFv was subsequently classified as Ab2γ type. Moreover, our results also demonstrated firstly that the anti-idiotype scFv could be used for pharmacokinetic measurement of circulating residual antibody in lymphoma patients treated with chimera anti-CD22 monoclonal antibody SM03. Of important, the present approach could be easily adopted to generate anti-idiotype antibodies for therapeutic antibodies targeting membrane proteins, saving the cost and time for producing a soluble antigen.

Generation of Anti-Idiotype scFv for Pharmacokinetic Measurement in Lymphoma Patients Treated with Chimera Anti-CD22 Antibody SM03

PubMed Central

Zhao, Qi; Wong, Pui-Fan; Lee, Susanna S. T.; Leung, Shui-On; Cheung, Wing-Tai; Wang, Jun-Zhi

2014-01-01

Pre-clinical and clinical studies of therapeutic antibodies require highly specific reagents to examine their immune responses, bio-distributions, immunogenicity, and pharmacodynamics in patients. Selective antigen-mimicking anti-idiotype antibody facilitates the assessment of therapeutic antibody in the detection, quantitation and characterization of antibody immune responses. Using mouse specific degenerate primer pairs and splenocytic RNA, we generated an idiotype antibody-immunized phage-displayed scFv library in which an anti-idiotype antibody against the therapeutic chimera anti-CD22 antibody SM03 was isolated. The anti-idiotype scFv recognized the idiotype of anti-CD22 antibody and inhibited binding of SM03 to CD22 on Raji cell surface. The anti-idiotype scFv was subsequently classified as Ab2γ type. Moreover, our results also demonstrated firstly that the anti-idiotype scFv could be used for pharmacokinetic measurement of circulating residual antibody in lymphoma patients treated with chimera anti-CD22 monoclonal antibody SM03. Of important, the present approach could be easily adopted to generate anti-idiotype antibodies for therapeutic antibodies targeting membrane proteins, saving the cost and time for producing a soluble antigen. PMID:24816427

Family influence: key to fruit and vegetable consumption among fourth- and fifth-grade students.

PubMed

Gross, Susan M; Pollock, Elizabeth Davenport; Braun, Bonnie

2010-01-01

To assess social and familial environmental influences on fruit and vegetable (FV) consumption of fourth- and fifth-graders living in a culturally diverse, urban setting. In 2006, students from 9 fourth- and fifth-grade classrooms from a public school in the Washington-Baltimore Metropolitan Region were recruited as part of the Food Stamp Nutrition Education's initiative to increase FV consumption among school-aged children in low-income schools. This intervention was conducted in partnership with the school. Ninety-three fourth- and fifth-grade students and their parents. The average daily FV consumption of elementary school students was the dependent variable. Home engagement (eg, student participation in menu planning and food selection at home), parental modeling, school environment, peer influences, and individual characteristics were the independent variables. Multiple regression analysis. Family and home environment factors explained more than 50% of the variance in students' FV consumption. Interventions designed to influence school-aged children's FV consumption should target family members and consider incorporating strategies to reach families and promote meal planning activities with the children in their homes. Copyright 2010 Society for Nutrition Education. Published by Elsevier Inc. All rights reserved.

Phage displayed scFv: pIII scaffold may fine tune binding specificity.

PubMed

Goswami, Pooja; Saini, Deepti; Sinha, Subrata

2009-10-01

The fine specificity of antibodies is important for their discriminating powers during diagnostics and in vivo therapy. We have attempted to isolate human scFv antibodies to the oncofetal antigen, the placental isozyme of alkaline phosphatase (PLAP) in which it is important to distinguish between the closely related intestinal alkaline phosphatase (IAP) and bone alkaline phosphatase (BAP) isozymes. As the antibodies are selected in the phage displayed form and might be finally used as different entities, including the soluble scFv form, it may be important to look at the influence of scaffolds in determining specificity. There have been earlier reports of the role of the constant region and other scaffolding proteins in determining specificity. In this paper, we report isolation of one such clone, E6, which showed specificity to PLAP in phage antibody form but lost the specificity when soluble scFv was tested for same, and showed partial cross reactivity to BAP. We suggest that the altered specificity of scFv might be the result of loss of phage pIII scaffold, which is present in phage-displayed antibody and may help the displayed antibody to assume specific conformational structure, which may govern binding characteristics of the same.

EMMPRIN reduction via scFv-M6-1B9 intrabody affects α3β1-integrin and MCT1 functions and results in suppression of progressive phenotype in the colorectal cancer cell line Caco-2.

PubMed

Sangboonruang, S; Thammasit, P; Intasai, N; Kasinrerk, W; Tayapiwatana, C; Tragoolpua, K

2014-06-01

Extracellular matrix metalloproteinase inducer (EMMPRIN) exhibits overexpression in various cancers and promotes cancer progression and metastasis via the interaction with its associated molecules. The scFv-M6-1B9 intrabody has a potential ability to reduce EMMPRIN cell surface expression. However, the subsequent effect of scFv-M6-1B9 intrabody-mediated EMMPRIN abatement on its related molecules, α3β1-integrin, MCT1, MMP-2 and MMP-9, is undefined. Our results demonstrated that the scFv-M6-1B9 intrabody efficiently decreased α3β1-integrin cell surface expression levels. In addition, intracellular accumulation of MCT1 and lactate were increased. These results lead to suppression of features characteristic for tumor progression, including cell migration, proliferation and invasion, in a colorectal cancer cell line (Caco-2) although there was no difference in MMP expression. Thus, EMMPRIN represents an attractive target molecule for the disruption of cancer proliferation and metastasis. An scFv-M6-1B9 intrabody-based approach could be relevant for cancer gene therapy.

Production of anti-amoxicillin ScFv antibody and simulation studying its molecular recognition mechanism for penicillins.

PubMed

Liu, Jing; Zhang, Hui C; Duan, Chang F; Dong, Jun; Zhao, Guo X; Wang, Jian P; Li, Nan; Liu, Jin Z; Li, Yu W

2016-11-01

The molecular recognition mechanism of an antibody for its hapten is very interesting. The objective of this research was to study the intermolecular interactions of an anti-amoxicillin antibody with penicillin drugs. The single chain variable fragment (ScFv) antibody was generated from a hybridoma cell strain excreting the monoclonal antibody for amoxicillin. The recombinant ScFv antibody showed similar recognition ability for penicillins to its parental monoclonal antibody: simultaneous recognizing 11 penicillins with cross-reactivities of 18-107%. The three-dimensional structure of the ScFv antibody was simulated by using homology modeling, and its intermolecular interactions with 11 penicillins were studied by using molecular docking. Results showed that three CDRs are involved in antibody recognition; CDR L3 Arg 100, CDR H3 Tyr226, and CDR H3 Arg 228 were the key contact amino acid residues; hydrogen bonding was the main antibody-drug intermolecular force; and the core structure of penicillin drugs was the main antibody binding position. These results could explain the recognition mechanism of anti-amoxicillin antibody for amoxicillin and its analogs. This is the first study reporting the production of ScFv antibody for penicillins and stimulation studying its recognition mechanism.

What Are Rare Clotting Factor Deficiencies?

MedlinePlus

... Deficiency Factor V Deficiency Combined FV & FVIII Deficiencies Factor VII Deficiency Factor X Deficiency Factor XI Deficiency Factor ... Deficiency Factor V Deficiency Combined FV & FVIII Deficiencies Factor VII Deficiency Factor X Deficiency Factor XI Deficiency Factor ...

Selection of Single-Chain Antibodies against the VP8* Subunit of Rotavirus VP4 Outer Capsid Protein and Their Expression in Lactobacillus casei

PubMed Central

Monedero, Vicente; Rodríguez-Díaz, Jesús; Viana, Rosa; Buesa, Javier; Pérez-Martínez, Gaspar

2004-01-01

Single-chain antibodies (scFv) recognizing the VP8* fraction of rotavirus outer capsid and blocking rotavirus infection in vitro were isolated by phage display. Vectors for the extracellular expression in Lactobacillus casei of one of the scFv were constructed. L. casei was able to secrete active scFv to the growth medium, showing the potential of probiotic bacteria to be engineered to express molecules suitable for in vivo antirotavirus therapies. PMID:15528568

Assimilation of MODIS and VIIRS AOD to improve aerosols forecasts with FV3-GOCART

NASA Astrophysics Data System (ADS)

Pagowski, M.

2017-12-01

In 2016 NOAA chose the FV3 dynamical core as a basis for its future global modeling system. We present an implementation of aerosol module in the FV3 model and its assimilation framework. The parameterization of aerosols is based on the GOCART scheme. The assimilation methodology relies on hybrid 3D-Var and EnKF methods. Aerosol observations include aerosol optical depth at 550 nm from VIIRS satellite. Results and evaluation of the system against independent observations and NASA's MERRA-2 is shown.

Tattoos

MedlinePlus

... Tattoos might make eczema, psoriasis, or other skin conditions flare up. Serious problems can happen if you try to do a tattoo yourself, have a friend do it for you, or have it done in any unclean environment. Skin infections caused by bacteria, viruses, or fungi can happen if the skin ...

Blocking monocyte transmigration in in vitro system by an anti-CD99 human antibody in single chain fragment variable (scFv) format. Efficient large scale purification of biological active scFv from inclusion bodies in E. coli expression system

PubMed Central

Moricoli, Diego; Muller, William A.; Carbonella, Damiano Cosimo; Balducci, Maria Cristina; Dominici, Sabrina; Fiori, Valentina; Watson, Richard; Weber, Evan; Cianfriglia, Maurizio; Scotlandi, Katia; Magnani, Mauro

2015-01-01

Migration of leukocytes into a site of inflammation involves several steps mediated by various families of adhesion molecules. CD99 play a significant role in transendothelial migration (TEM) of leukocytes. Inhibition of TEM by specific monoclonal antibody (mAb) can provide a potent therapeutic approach to treating inflammatory conditions. However, the therapeutic utilization of whole IgG can lead to an inappropriate activation of Fc receptor-expressing cells inducing serious adverse side effects due to cytokine release. In this regard, specific recombinant antibody in single chain variable fragments (scFvs) originated by phage library may offer a solution by affecting TEM function in a safe clinical context. However, this consideration requires large scale production of functional scFv antibodies under GMP conditions and hence, the absence of toxic reagents utilized for the solubilization and refolding steps of inclusion bodies that may discourage industrial application of these antibody fragments. In order to apply the scFv anti-CD99 named C7A in a clinical setting we herein describe an efficient and large scale production of the antibody fragments expressed in E.coli as insoluble protein avoiding gel filtration chromatography approach, and laborious refolding step pre- and post-purification. Using differential salt elution which is a simple, reproducible and effective procedure we are able to separate scFv in monomer format from aggregates. The purified scFv antibody C7A exhibits inhibitory activity comparable to an antagonistic conventional mAb, thus providing an excellent agent for blocking CD99 signalling. Thanks to the original purification protocol that can be extended to other scFvs that are expressed as inclusion bodies in bacterial systems, the scFv anti-CD99 C7A herein described represents the first step towards the construction of new antibody therapeutic. PMID:24798881

A cross-sectional descriptive study of the family functioning, health and social support of hospital patients with family violence backgrounds.

PubMed

Kivelä, Salla; Leppäkoski, Tuija; Helminen, Mika; Paavilainen, Eija

2017-12-19

Family violence (FV) has serious effects on the health and well-being of the family. The health sector plays a vital role in FV prevention by helping to identify abuse early, providing victims with the necessary treatment and referring patients to appropriate care. The aim of the present cross-sectional study was to describe the prevalence of FV among hospital patients, as well as to assess the association between family functioning, health and social support, considering patients as the perpetrators or victims of violence. The data were collected using a Family Functioning, Health and Social Support (FAFHES) questionnaire that was given to patients who visited a Finnish central hospital between October 2012 and April 2013. As a result, the data (N = 188) were contributed by the patients who returned the questionnaire and gave permission for a follow-up survey. The participation rate was 47%, of which 73% were women and 27% were men. Their ages ranged from 18 to 89 years. The data were analysed with quantitative methods using the unadjusted analyses and linear regression model. In total, 24% of both the male and female participants had experienced or used violence at home or in the family. Of these, 22 had been the perpetrators, and 23 had been the victims. Participants in relationships and who were living together had less violence than singles and those who were not living together. The family functioning and health of the participants who had experienced or used FV were worse than those of the participants who had not. Various patients can have an FV background, and nursing professionals are on the front line to identify and intervene in FV situations. The results of this study can be utilised in the treatment of FV victims and perpetrators by training healthcare workers to identify and intervene in violence. © 2017 Nordic College of Caring Science.

Assessment and monitoring of flow limitation and other parameters from flow/volume loops.

PubMed

Dueck, R

2000-01-01

Flow/volume (F/V) spirometry is routinely used for assessing the type and severity of lung disease. Forced vital capacity (FVC) and timed vital capacity (FEV1) provide the best estimates of airflow obstruction in patients with asthma, chronic obstructive pulmonary disease (COPD) and emphysema. Computerized spirometers are now available for early home recognition of asthma exacerbation in high risk patients with severe persistent disease, and for recognition of either infection or rejection in lung transplant patients. Patients with severe COPD may exhibit expiratory flow limitation (EFL) on tidal volume (VT) expiratory F/V (VTF/V) curves, either with or without applying negative expiratory pressure (NEP). EFL results in dynamic hyperinflation and persistently raised alveolar pressure or intrinsic PEEP (PEEPi). Hyperinflation and raised PEEPi greatly enhance dyspnea with exertion through the added work of the threshold load needed to overcome raised pleural pressure. Esophageal (pleural) pressure monitoring may be added to VTF/V loops for assessing the severity of PEEPi: 1) to optimize assisted ventilation by mask or via endotracheal tube with high inspiratory flow rates to lower I:E ratio, and 2) to assess the efficacy of either pressure support ventilation (PSV) or low level extrinsic PEEP in reducing the threshold load of PEEPi. Intraoperative tidal volume F/V loops can also be used to document the efficacy of emphysema lung volume reduction surgery (LVRS) via disappearance of EFL. Finally, the mechanism of ventilatory constraint can be identified with the use of exercise tidal volume F/V loops referenced to maximum F/V loops and static lung volumes. Patients with severe COPD show inspiratory F/V loops approaching 95% of total lung capacity, and flow limitation over the entire expiratory F/V curve during light levels of exercise. Surprisingly, patients with a history of congestive heart failure may lower lung volume towards residual volume during exercise, thereby reducing airway diameter and inducing expiratory flow limitation.

In vitro activity of monoclonal and recombinant yeast killer toxin-like antibodies against antibiotic-resistant gram-positive cocci.

PubMed Central

Conti, S.; Magliani, W.; Arseni, S.; Dieci, E.; Frazzi, R.; Salati, A.; Varaldo, P. E.; Polonelli, L.

2000-01-01

BACKGROUND: Monoclonal (mAbKT) and recombinant single-chain (scFvKT) anti-idiotypic antibodies were produced to represent the internal image of a yeast killer toxin (KT) characterized by a wide spectrum of antimicrobial activity, including gram-positive cocci. Pathogenic eukaryotic and prokaryotic microorganisms, such as Candida albicans, Pneumocystis carinii, and a multidrug-resistant strain of Mycobacterium tuberculosis, presenting specific, although yet undefined, KT-cell wall receptors (KTR), have proven to be killed in vitro by mAbKT and scFvKT. mAbKT and scFvKT exert a therapeutic effect in vivo in experimental models of candidiasis and pneumocystosis by mimicking the functional activity of protective antibodies naturally produced in humans against KTR of infecting microorganisms. The swelling tide of concern over increasing bacterial resistance to antibiotic drugs gives the impetus to develop new therapeutic compounds against microbial threat. Thus, the in vitro bactericidal activity of mAbKT and scFvKT against gram-positive, drug-resistant cocci of major epidemiological interest was investigated. MATERIALS AND METHODS: mAbKT and scFvKT generated by hybridoma and DNA recombinant technology from the spleen lymphocytes of mice immunized with a KT-neutralizing monoclonal antibody (mAb KT4) were used in a conventional colony forming unit (CFU) assay to determine, from a qualitative point of view, their bactericidal activity against Staphylococcus aureus, S. haemolyticus, Enterococcus faecalis, E. faecium, and Streptococcus pneumoniae strains. These bacterial strains are characterized by different patterns of resistance to antibiotics, including methicillin, vancomycin, and penicillin. RESULTS: According to the experimental conditions adopted, no bacterial isolate proved to be resistant to the activity of mAbKT and scFvKT. CONCLUSIONS: scFvKT exerted a microbicidal activity against multidrug resistant bacteria, which may represent the basis for the drug modeling of new antibiotics with broad antibacterial spectra to tackle the emergence of microbial resistance. PMID:10997342

Improvement in Fruit and Vegetable Consumption Associated with More Favorable Energy Density and Nutrient and Food Group Intake, but not Kilocalories.

PubMed

Thompson, Debbe; Ferry, Robert J; Cullen, Karen W; Liu, Yan

2016-09-01

Children generally do not consume adequate amounts of fruits and vegetables (F/V). Eating more F/V can improve energy density and overall diet quality. Our aim was to investigate whether improvements in F/V consumption were associated with improvements in energy density, total calories, and dietary components related to F/V. We performed secondary analyses of dietary data from a successful four-group randomized controlled trial promoting F/V. Data were collected at baseline, immediately after gameplay, and 3 months post intervention. Preadolescent child-parent dyads (n=400) were recruited. Eligibility criteria were 4th- or 5th-grade child (approximately 9 to 11 years old) with Internet access and a parent willing to participate in the intervention. Complete dietary data were collected on 387 of the 400 child participants. The videogame was available online on a secure, password-protected website. Dietary intake was assessed with three unannounced dietary recalls collected at each data-collection period via telephone by trained staff using Nutrition Data System for Research software. Energy density and F/V, nutrient, and food consumption were calculated. A 4×3 (group by time) repeated measures analysis of covariance with mixed-effect linear models was used. Covariates included child's sex, race/ethnicity, and total energy intake as well as parent's age and household education. Energy was excluded as a covariate in the energy density and energy models. Significant changes occurred in energy density. A significant interaction (group by time) was observed (F6, 515=2.40; P<0.05) in energy density from food only, while a significant time effect was observed for energy density from all foods and beverages (F2, 388=13.75; P<0.0001). Desirable changes were also observed in F/V-related dietary components. Increasing F/V consumption improved energy density and diet quality considerably in preadolescent children. Copyright © 2016 Academy of Nutrition and Dietetics. Published by Elsevier Inc. All rights reserved.

Replication-Competent Foamy Virus Vaccine Vectors as Novel Epitope Scaffolds for Immunotherapy

PubMed Central

Lei, Janet; Osen, Wolfram; Gardyan, Adriane; Hotz-Wagenblatt, Agnes; Wei, Guochao; Gissmann, Lutz; Eichmüller, Stefan; Löchelt, Martin

2015-01-01

The use of whole viruses as antigen scaffolds is a recent development in vaccination that improves immunogenicity without the need for additional adjuvants. Previous studies highlighted the potential of foamy viruses (FVs) in prophylactic vaccination and gene therapy. Replication-competent FVs can trigger immune signaling and integrate into the host genome, resulting in persistent antigen expression and a robust immune response. Here, we explored feline foamy virus (FFV) proteins as scaffolds for therapeutic B and T cell epitope delivery in vitro. Infection- and cancer-related B and T cell epitopes were grafted into FFV Gag, Env, or Bet by residue replacement, either at sites of high local sequence homology between the epitope and the host protein or in regions known to tolerate sequence alterations. Modified proviruses were evaluated in vitro for protein steady state levels, particle release, and virus titer in permissive cells. Modification of Gag and Env was mostly detrimental to their function. As anticipated, modification of Bet had no impact on virion release and affected virus titers of only some recombinants. Further evaluation of Bet as an epitope carrier was performed using T cell epitopes from the model antigen chicken ovalbumin (OVA), human tyrosinase-related protein 2 (TRP-2), and oncoprotein E7 of human papillomavirus type 16 (HPV16E7). Transfection of murine cells with constructs encoding Bet-epitope chimeric proteins led to efficient MHC-I-restricted epitope presentation as confirmed by interferon-gamma enzyme-linked immunospot assays using epitope-specific cytotoxic T lymphocyte (CTL) lines. FFV infection-mediated transduction of cells with epitope-carrying Bet also induced T-cell responses, albeit with reduced efficacy, in a process independent from the presence of free peptides. We show that primate FV Bet is also a promising T cell epitope carrier for clinical translation. The data demonstrate the utility of replication-competent and -attenuated FVs as antigen carriers in immunotherapy. PMID:26397953

Decreased Flight Activity in Culex pipiens (Diptera: Culicidae) Naturally Infected With Culex flavivirus.

PubMed

Newman, Christina M; Anderson, Tavis K; Goldberg, Tony L

2016-01-01

Insect-specific flaviviruses (ISFVs) commonly infect vectors of mosquito-borne arboviruses. To investigate whether infection with an ISFV might affect mosquito flight behavior, we quantified flight behavior in Culex pipiens L. naturally infected with Culex flavivirus (CxFV). We observed a significant reduction in the scotophase (dark hours) flight activity of CxFV-positive mosquitoes relative to CxFV-negative mosquitoes, but only a marginal reduction in photophase (light hours) flight activity, and no change in the circadian pattern of flight activity. These results suggest that CxFV infection alters the flight activity of naturally infected Cx. pipiens most dramatically when these vectors are likely to be host seeking and may therefore affect the transmission of medically important arboviruses. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Spin wave propagation in perpendicular magnetized 20 nm Yttrium Iron Garnet with different antenna design

NASA Astrophysics Data System (ADS)

Chen, Jilei; Stueckler, Tobias; Zhang, Youguang; Zhao, Weisheng; Yu, Haiming; Chang, Houchen; Liu, Tao; Wu, Mingzhong; Liu, Chuanpu; Liao, Zhimin; Yu, Dapeng; Fert Beijing research institute Team; Colorado State University Team; Peking University Collaboration

Magnonics offers a new way to transport information using spin waves free of charge current and could lead to a new paradigm in the area of computing. Forward volume (FV) mode spin wave with perpendicular magnetized configuration is suitable for spin wave logic device because it is free of non-reciprocity effect. Here, we study FV mode spin wave propagation in YIG thin film with an ultra-low damping. We integrated differently designed antenna i.e., coplanar waveguide and micro stripline with different dimensions. The k vectors of the spin waves defined by the design of the antenna are calculated using Fourier transform. We show FV mode spin wave propagation results by measuring S12 parameter from vector network analyzer and we extract the group velocity of the FV mode spin wave as well as its dispersion relations.

A Jacalin-Related Lectin Regulated the Formation of Aerial Mycelium and Fruiting Body in Flammulina velutipes

PubMed Central

Lu, Yuan-Ping; Chen, Ren-Liang; Long, Ying; Li, Xiao; Jiang, Yu-Ji; Xie, Bao-Gui

2016-01-01

Flammulina velutipes, one of the most popular mushroom species in the world, has been recognized as a useful model system to study the biochemical and physiological aspects of the formation and elongation of fruit body. However, few reports have been published on the regulation of fruiting body formation in F. velutipes at the molecular level. In this study, a jacalin-related lectin gene from F. velutipes was characterized. The phylogenetic tree revealed that Fv-JRL1 clustered with other basidiomycete jacalin-like lectins. Moreover, the transcriptional pattern of the Fv-JRL1 gene in different developmental stages of F. velutipes implied that Fv-JRL1 could be important for formation of fruit body. Additionally, RNA interference (RNAi) and overexpression analyses provided powerful evidence that the lectin gene Fv-JRL1 from F. velutipes plays important roles in fruiting body formation. PMID:27916794

In Vivo Antiviral Activity of 1,3-Bis(2-Chloroethyl)-1-Nitrosourea

PubMed Central

Sidwell, Robert W.; Dixon, Glen J.; Sellers, Sara M.; Schabel, Frank M.

1965-01-01

A prolongation in the lives of Swiss mice inoculated intracerebrally with lymphocytic choriomeningitis virus (LCM) was observed after treatment with 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU). A variety of treatment schedules, including therapy once or twice daily up to 17 days and single treatments at various times after virus inoculation, were employed. Virus titers ranging to greater than 104 were detected in the blood and brains of surviving drug-treated animals. In three comparative studies in which different treatment schedules were used, BCNU was shown to exert a protective effect approximately equal to that of methotrexate in LCM virus-infected mice. Tests were also carried out to investigate the activity of BCNU in mice experimentally infected with eastern equine encephalomyelitis (EEE) virus, western equine encephalomyelitis virus, Semliki Forest (SF) virus, herpes simplex virus, influenza virus strain PR8, vaccinia virus strain WR, Rous sarcoma virus, Friend leukemia virus (FLV), and poliovirus. Slight increases in life span were observed in the treated EEE, SF, and influenza PR8 virus-infected animals. Significant reduction in splenomegaly in FLV-infected animals treated with BCNU was demonstrated. The possible mechanisms of LCM virus inhibition by BCNU, on the basis of these and other studies, were postulated to be either specific antiviral activity or inhibition of “lethal” immune response to the LCM virus. Each of these postulates is discussed. PMID:14339266

Genetic control of T cell responsiveness to the Friend murine leukemia virus envelope antigen. Identification of class II loci of the H-2 as immune response genes

PubMed Central

1988-01-01

T cells primed specifically for the envelope glycoprotein of Friend murine leukemia helper virus (F-MuLV) were prepared by immunizing mice with a recombinant vaccinia virus that expressed the entire env gene of F-MuLV. Significant proliferative responses of F-MuLV envelope- specific, H-2a/b T cells were observed when the T cells were stimulated with antigen-pulsed peritoneal exudate cells (PEC) having the b allele at the K, A beta, A alpha, and E beta loci of the H-2. On the other hand, PEC having only the kappa allele at these loci did not induce the envelope-specific T cell proliferation, even when the PEC had the b allele at the E alpha, S, or D loci. F-MuLV envelope-specific proliferation of H-2a/b T cells under the stimulation of antigen- pulsed, H-2a/b PEC was specifically blocked with anti-I-Ab and anti-I- Ek mAbs but not with anti-Kb, anti-Kk, or anti-I-Ak mAbs. Moreover, (B10.MBR x A/WySn)F1 mice that have the b allele only at the K locus but not in I-A subregion were nonresponders to the envelope glycoprotein, and the bm12 mutation at the A beta locus completely abolished the T cell responsiveness to this antigen. These results indicate that proliferative T cells recognize a limited number of epitopes on F-MuLV envelope protein in the context of I-Ab, hybrid I- Ak/b, and/or hybrid I-Ek/b class II MHC molecules but fail to recognize the same envelope protein in the context of I-Ak or I-Ek molecules. This influence of the H-2I region on T cell recognition of the envelope glycoprotein appeared to control in vivo induction of protective immunity against Friend virus complex after immunization with the vaccinia-F-MuLV env vaccine. Thus, these results provide, for the first time, direct evidence for Ir gene-controlled responder/nonresponder phenotypes influencing the immune response to a pathogenic virus of mice. PMID:3141552

Friend leukemia virus transformed cells exposed to microgravity in the presence of DMSO (7-IML-1)

NASA Technical Reports Server (NTRS)

Cogoli, Augusto

1992-01-01

The purpose of this experiment is to study the adaptation of living cells to microgravity. The in vitro transformation of Friend cells by Dimethylsufoxide (DMSO) is a good model for the study of cell differentiation and protein biosynthesis. Cultures of cells will be prepared shortly before launch. Once in space, transformation will be induced by injection of DMSO. One set of cultures will be chemically fixed with glutaraldehyde for electron microscope investigations; another set will be preserved for determining the amount of hemogloben produced and the extent of cell proliferation.

Diverse binding site structures revealed in homology models of polyreactive immunoglobulins

NASA Astrophysics Data System (ADS)

Ramsland, Paul A.; Guddat, Luke W.; Edmundson, Allen B.; Raison, Robert L.

1997-09-01

We describe here computer-assisted homology models of the combiningsite structure of three polyreactive immunoglobulins. Template-based modelsof Fv (VL-VH) fragments were derived forthe surface IgM expressed by the malignant CD5 positive B cells from threepatients with chronic lymphocytic leukaemia (CLL). The conserved frameworkregions were constructed using crystal coordinates taken from highlyhomologous human variable domain structures (Pot and Hil). Complementaritydetermining regions (CDRs) were predicted by grafting loops, taken fromknown immunoglobulin structures, onto the Fv framework models. The CDRtemplates were chosen, where possible, to be of the same length and of highresidue identity or similarity. LCDR1, 2 and 3 as well as HCDR1 and 2 forthe Fv were constructed using this strategy. For HCDR3 prediction, adatabase containing the Cartesian coordinates of 30 of these loops wascompiled from unliganded antibody X-ray crystallographic structures and anHCDR3 of the same length as that of the B CLL Fv was selected as a template.In one case (Yar), the resulting HCDR3 model gave unfavourable interactionswhen incorporated into the Fv model. This HCDR3 was therefore modelled usingan alternative strategy of construction of the loop stems, using apreviously described HCDR3 conformation (Pot), followed by chain closurewith a β-turn. The template models were subjected to positionalrefinement using energy minimisation and molecular dynamics simulations(X-PLOR). An electrostatic surface description (GRASP) did not reveal acommon structural feature within the binding sites of the three polyreactiveFv. Thus, polyreactive immunoglobulins may recognise similar and multipleantigens through a diverse array of binding site structures.

Effects of Self-Efficacy on Healthy Eating Depends on Normative Support: a Prospective Study of Long-Haul Truck Drivers.

PubMed

Hamilton, Kyra; Hagger, Martin S

2018-04-01

Fruit and vegetable intake (FV) is insufficient in industrialized nations and there is excess of discretionary food choices (DC; foods high in fat, sugar, and salt). Long-haul truck drivers are considered a particularly at-risk group given the limited food choices and normatively reinforced eating habits at truck rest-stops. Self-efficacy and normative support are key determinants of eating behavior yet the processes underlying their effects on behavior are not well understood. We tested the direct and interactive effects of self-efficacy and normative support on healthy eating behaviors in long-haul truck drivers in a prospective correlational study. Long-haul truck drivers (N = 82) completed an initial survey containing self-report measures of behavioral intentions, perceived normative support, and self-efficacy for their FV and DC behaviors. Participants completed a follow-up survey 1 week later in which they self-reported their FV and DC behavior. A mediated moderation analysis identified an interactive effect of self-efficacy and normative support on behavior mediated by intention for FV and DC behavior. Specifically, we confirmed a compensation effect in which self-efficacy was more likely to have an effect on FV and DC behavior through intentions in participants with low normative support. Results indicate the importance of self-efficacy in predicting FV and DC intentions and behavior in the absence of a supportive normative environment. The compensatory effect of self-efficacy beliefs on behavior through intentions when normative support is low should be confirmed using experimental methods.