Effect of packaging atmospheres on storage quality characteristics of heavily marbled beef longissimus steaks.

PubMed

Yang, Xiaoyin; Zhang, Yimin; Zhu, Lixian; Han, Mingshan; Gao, Shujuan; Luo, Xin

2016-07-01

The objective of this study was to investigate the effects of modified atmosphere packaging (MAP) systems on shelf-life and quality of beef steaks with high marbling. Four packaging types were used including 80% O2 MAP (80% O2+20% CO2), 50% O2 MAP (50% O2+30% CO2+20% N2), carbon monoxide MAP (0.4% CO+30% CO2+69.6% N2) and vacuum packaging (VP). Steaks were displayed under simulated retail conditions at 4°C for 12days. Purge loss, pH, color stability, oxidative stability and microbial counts were monitored. Aerobically packaged steaks exhibited a bright-red color at the first 4days. However, discoloration and oxidation became major factors limiting their shelf-life to 8days. Compared with aerobic packaging, anaerobic packaging extended shelf-life of heavily marbled beef steaks, due to better color stability, together with lower oxidation and microbial populations. Among all packaging methods, CO-MAP had the best preservation for steaks, with more red color than other packaging types. Copyright © 2016 Elsevier Ltd. All rights reserved.

Super-chilling (-0.7°C) with high-CO2 packaging inhibits biochemical changes of microbial origin in catfish (Clarias gariepinus) muscle during storage.

PubMed

Zhu, Yingchun; Ma, Lizhen; Yang, Hua; Xiao, Yan; Xiong, Youling L

2016-09-01

Controlled freezing-point storage (CFPS) is an emerging preservative technique desirable for fish. In the present study, catfish fillets were stored at -0.7°C under different packaging atmospheres: air (AP), vacuum (VP), and 60% CO2/40% N2 (MAP). Chemical, microbiological, and sensory analyses were performed during storage. Results showed the following descending order of chemical changes (degradation of nucleotides, conversion of protein to volatile-based nitrogen and biogenic amines, and production of trimethylamine nitrogen), as well as loss of sensory properties: 4°C AP>-0.7°C AP≈4°C VP>-0.7°C VP≈4°C MAP>-0.7°C MAP. The chemical changes were well-correlated with microbial growth suggesting the microbiological pathways. Hence, CFPS at -0.7°C in combination with high-CO2 MAP can effectively maintain the quality of fresh catfish meat compared to traditional preservation methods. Copyright © 2016 Elsevier Ltd. All rights reserved.

Culture-dependent and culture-independent assessment of spoilage community growth on VP lamb meat from packaging to past end of shelf-life.

PubMed

Kaur, Mandeep; Shang, Hongshan; Tamplin, Mark; Ross, Tom; Bowman, John P

2017-12-01

Packaging and storage temperature are important factors that influence the shelf-life of vacuum packed (VP) meat. In this study the shelf-life of VP bone-in lamb hind shanks stored at 8 °C and -1.2 °C was determined in parallel to analyses of starting and eventual spoilage bacterial communities via Illumina MiSeq based 16S rRNA amplicon sequencing. The mean total viable counts (TVC) and lactic acid bacterial viable counts (LAB) were observed to increase to log 7.5 CFU/cm 2 and 7 CFU/cm 2 after 6 and 42 days at 8 °C and -1.2 °C and stayed stable until shelf-life termination after 13 and 124 days, respectively. The sequence data showed initial communities were patchily distributed and were mainly derived from skin microbiome taxa likely prevalent within the abattoir. A broad diversity of VP meat associated specific spoilage organisms (SSO) were comparatively abundant in this initial population. Overtime meat spoilage communities developed a distinctive and stable microbiome. At -1.2 °C SSO included mainly Carnobacterium, Yersinia and Clostridium spp. while at 8 °C SSO expanded to include Hafnia, Lactococcus, Providencia spp. Growth curves inferred from the sequence data after taking into account rRNA copy number suggested that SSO growth rates were consistent with overall growth rates determined from TVC and LAB data and are predictable. Crown Copyright © 2017. Published by Elsevier Ltd. All rights reserved.

Mechanism for Coordinated RNA Packaging and Genome Replication by Rotavirus Polymerase VP1

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

Lu, Xiaohui; McDonald, Sarah M.; Tortorici, M. Alejandra

2009-04-08

Rotavirus RNA-dependent RNA polymerase VP1 catalyzes RNA synthesis within a subviral particle. This activity depends on core shell protein VP2. A conserved sequence at the 3' end of plus-strand RNA templates is important for polymerase association and genome replication. We have determined the structure of VP1 at 2.9 {angstrom} resolution, as apoenzyme and in complex with RNA. The cage-like enzyme is similar to reovirus {lambda}3, with four tunnels leading to or from a central, catalytic cavity. A distinguishing characteristic of VP1 is specific recognition, by conserved features of the template-entry channel, of four bases, UGUG, in the conserved 3' sequence.more » Well-defined interactions with these bases position the RNA so that its 3' end overshoots the initiating register, producing a stable but catalytically inactive complex. We propose that specific 3' end recognition selects rotavirus RNA for packaging and that VP2 activates the autoinhibited VP1/RNA complex to coordinate packaging and genome replication.« less

Formation of protein sub-visible particles during vacuum degassing of etanercept solutions.

PubMed

Wang, Haibin; Zheng, Hong-Jian; Wang, Zhao; Bai, Hua; Carpenter, John F; Chen, Shuqing; Fang, Wei-Jie

2014-05-01

The main purpose of this manuscript is to describe a phenomenon in which vacuum degassing a reconstituted freeze-dried fusion protein etanercept formulation caused a significant amount of protein sub-visible particles (SbVP). Physical stability of etanercept was monitored by micro-flow imaging (MFI), dynamic light scattering (DLS), size-exclusion high pressure liquid chromatography (SE-HPLC) and far- and near-ultraviolet circular dichroism (far- and near-UV CD). One potential explanation of this phenomenon is that bubble collapses when the vacuum is applied, leads to substantial heat formation, and ultimately free radical formation. Subsequently, the effect of a free-radical scavenger (ascorbic acid, AA) on SbVP formation was also evaluated. Degassing of etanercept solution by applying vacuum caused substantial increase of SbVP, as detected by MFI and DLS. However, traditional techniques such as SE-HPLC could not detect any change. The addition of free-radical scavenger had minimal effect on SbVP formation, therefore the formation of free radicals was probably not the main cause for this effect. Copyright © 2014 Elsevier B.V. All rights reserved.

Color stability of ground beef packaged in a low carbon monoxide atmosphere or vacuum.

PubMed

Jeong, Jong Youn; Claus, James R

2011-01-01

Ground beef was either packaged in an atmosphere of 0.4% CO, 30% CO₂, and 69.6% N₂ (CO-MAP) or vacuum. After storage (48 h, 2-3°C), packages of CO-MAP and vacuum were opened and overwrapped with polyvinyl chloride. Other CO-MAP and vacuum packages were left intact. Packages were initially displayed for 7 days (2-3°C). Intact packages were further displayed up to 35 days before being opened and displayed (1 or 3 days). Intact CO-MAP packaged ground beef was always more red than intact vacuum-packaged ground beef. Color was relatively stable for both types of intact packages over 35 days of display. Upon opening CO-MAP packaged ground beef, the red color decreased slower than in ground beef from vacuum packages. Published by Elsevier Ltd.

Ebola virus VP24 interacts with NP to facilitate nucleocapsid assembly and genome packaging.

PubMed

Banadyga, Logan; Hoenen, Thomas; Ambroggio, Xavier; Dunham, Eric; Groseth, Allison; Ebihara, Hideki

2017-08-09

Ebola virus causes devastating hemorrhagic fever outbreaks for which no approved therapeutic exists. The viral nucleocapsid, which is minimally composed of the proteins NP, VP35, and VP24, represents an attractive target for drug development; however, the molecular determinants that govern the interactions and functions of these three proteins are still unknown. Through a series of mutational analyses, in combination with biochemical and bioinformatics approaches, we identified a region on VP24 that was critical for its interaction with NP. Importantly, we demonstrated that the interaction between VP24 and NP was required for both nucleocapsid assembly and genome packaging. Not only does this study underscore the critical role that these proteins play in the viral replication cycle, but it also identifies a key interaction interface on VP24 that may serve as a novel target for antiviral therapeutic intervention.

The impact of enterocin AS-48 on the shelf-life and safety of sardines (Sardina pilchardus) under different storage conditions.

PubMed

Ananou, S; Zentar, H; Martínez-Bueno, M; Gálvez, A; Maqueda, M; Valdivia, E

2014-12-01

The purpose of this study was to determine the effect of enterocin AS-48, packaged under normal atmosphere (NA), vacuum (VP) or modified atmosphere (MAP) on the shelf life and safety of fresh sardines (Sardina pilchardus) stored at 5 °C. We studied the effect of these hurdles, alone or combined, on the relevant autochthonous bacterial populations. Total volatile basic nitrogen (TVB-N) content was used as indicative of freshness. Levels of biogenic amines cadaverine, putrescine, tyramine, and histamine were also determined. The application of AS-48 did not reduce the mesophilic, psychrotrophic, or Gram negative bacteria viable cell counts under any of the storage conditions tested. AS-48 did cause significant reductions in viable staphylococci counts, especially under VP. In sardines under NA treated with AS-48, the populations of histamine- and tyramine-forming total and lactic acid bacteria (LAB) showed no significant reductions. MAP or VP with AS-48 allowed reductions (significant at some storage times) in histamine- and tyramine-forming LAB. The TVB-N content was also reduced under normal atmosphere and, especially, in sardines stored under MAP. The most interesting results are those concerning the decrease (by several fold) in the levels of the biogenic amines cadaverine, putrescine, tyramine, and histamine determined after treatment with AS-48. Copyright © 2014 Elsevier Ltd. All rights reserved.

Effects of Gas Composition in the Modified Atmosphere Packaging on the Shelf-life of Longissimus dorsi of Korean Native Black Pigs-Duroc Crossbred during Refrigerated Storage

PubMed Central

Muhlisin; Panjono; Kim, Dong Soo; Song, Yeong Rae; Lee, Sung-Jin; Lee, Jeong Koo; Lee, Sung Ki

2014-01-01

This study was conducted to observe the effects of gas composition in modified atmosphere packaging (MAP) on the shelf-life of Longissimus dorsi of Korean Native Black Pigs-Duroc Crossbred (KNP×D) during refrigerated storage. Muscle sample was obtained from the left side of carcass of seven months old of KNP×D barrow. The sample was sliced into 1 cm in thickness, placed on trays (two slices/tray) and filled with different gas composition, i.e. 0:20:80/O2:CO2:N2 (MAP1), 30:20:50/O2:CO2:N2 (MAP2) and 70:20:10/O2:CO2:N2 (MAP3). Other slices of sample were vacuum packed (VP) as a control. All packs were stored at 5±1°C. At 12 d of storage, pH value of MAP2 and MAP3 were higher (p<0.05) than that of MAP1 and pH value of MAP1 was higher (p<0.05) than that of VP. At 6 d of storage, redness (a*) value of MAP2 and MAP3 were higher (p<0.05) than that of VP and MAP1 and, at 9 and 12 d of storage, redness value of MAP3 was higher (p<0.05) than that of VP, MAP1, and MAP2. At 3, 6, 9, and 12 d of storage, the 2-thiobarbituric acid reactive substances (TBARS) value of MAP3 was higher than that of MAP2 and TBARS value of MAP2 was higher than that of VP and MAP1. At 3, 6, 9, and 12 d of storage, volatile basic nitrogen values of MAP2 and MAP3 were higher (p<0.05) than those of VP and MAP1. At 3 d of storage, total aerobic plate counts of MAP2 and MAP3 were higher (p<0.05) than those of VP and MAP1 and, at 6 d of storage, total aerobic plate counts of MAP3 was higher (p<0.05) than that of MAP1 and MAP2. However, there was no significant different total aerobic plate count among MAP1, MAP2, and MAP3 at 9 and 12 d of storage. There was no significant different total anaerobic plate count among MAP1, MAP2, and MAP3 during storage. It is concluded that the MAP containing 30:20:50/O2:CO2:N2 gas composition (MAP2) might be ideal for better meat quality for KNP×D meat. PMID:25083110

A highly miniaturized vacuum package for a trapped ion atomic clock

DOE PAGES

Schwindt, Peter D. D.; Jau, Yuan-Yu; Partner, Heather; ...

2016-05-12

We report on the development of a highly miniaturized vacuum package for use in an atomic clock utilizing trapped ytterbium-171 ions. The vacuum package is approximately 1 cm 3 in size and contains a linear quadrupole RF Paul ion trap, miniature neutral Yb sources, and a non-evaporable getter pump. We describe the fabrication process for making the Yb sources and assembling the vacuum package. To prepare the vacuum package for ion trapping, it was evacuated, baked at a high temperature, and then back filled with a helium buffer gas. Once appropriate vacuum conditions were achieved in the package, the packagemore » was sealed with a copper pinch-off and was then pumped only by the non-evaporable getter. We demonstrated ion trapping in this vacuum package and the operation of an atomic clock, stabilizing a local oscillator to the 12.6 GHz hyperfine transition of 171Yb +. The fractional frequency stability of the clock was measured to be 2 × 10 -11 / τ 1/2.« less

Herpesvirus capsid assembly and DNA packaging

PubMed Central

Heming, Jason D.; Conway, James F.; Homa, Fred L.

2017-01-01

Herpes simplex virus type I (HSV-1) is the causative agent of several pathologies ranging in severity from the common cold sore to life-threatening encephalitic infection. During productive lytic infection, over 80 viral proteins are expressed in a highly regulated manner, resulting in the replication of viral genomes and assembly of progeny virions. The virion of all herpesviruses consists of an external membrane envelope, a proteinaceous layer called the tegument, and an icosahedral capsid containing the double-stranded linear DNA genome. The capsid shell of HSV-1 is built from four structural proteins: a major capsid protein, VP5, which forms the capsomers (hexons and pentons), the triplex consisting of VP19C and VP23 found between the capsomers, and VP26 which binds to VP5 on hexons but not pentons. In addition, the dodecameric pUL6 portal complex occupies one of the 12 capsid vertices, and the capsid vertex specific component (CVSC), a heterotrimer complex of pUL17, pUL25 and pUL36 binds specifically to the triplexes adjacent to each penton. The capsid is assembled in the nucleus where the viral genome is packaged into newly assembled closed capsid shells. Cleavage and packaging of replicated, concatemeric viral DNA requires the seven viral proteins encoded by the UL6, UL15, UL17, UL25, UL28, UL32, and UL33 genes. Considerable advances have been made in understanding the structure of the herpesvirus capsid and the function of several of the DNA packaging proteins by applying biochemical, genetic, and structural techniques. This review is a summary of recent advances with respect to the structure of the HSV-1 virion capsid and what is known about the function of the seven packaging proteins and their interactions with each other and with the capsid shell. PMID:28528442

Effect of starter cultures and packaging methods on amino acid profile and eating quality characteristics of pork ham.

PubMed

Gogoi, Protiva; Borpuzari, R N; Borpuzari, T; Hazarika, R A; Bora, J R

2015-08-01

Wet cured pork hams were inoculated with a mixed starter cultures comprising of Lactobacillus acidophilus and Micrococcus varians M483 at the dose level of 106 cfu/g and the un inoculated hams served as controls. The amino acid profile of hams of the treated and the control groups stored at 4oC under MAP and VP and evaluated on 60th day of storage revealed that treated hams liberated higher concentration of free amino acids except for proline and methionine which were found in higher concentration (P < 0.01) in the MAP control samples. The MAP control samples liberated glutamic acid (85.65 ± 1.40 ppm), cystine (21.56 ± 1.14 ppm) and tyrosine (16.63 ± 1.94 ppm) whereas, the treated samples did not release these amino acids. The VP control samples too liberated cystine (6.98 ± 1.36 ppm) and arginine (42.70 ± 2.78 ppm) but the treated ham of the VP did not liberate these amino acids. The VP hams had higher concentration (P < 0.01) of free proline, glycine, alanine, valine, methionine, isoleucine, phenylalanine, lysine and histidine than the MAP samples. Colour analysis of ham using CIE Lab colour system revealed that the treated samples had significantly higher concentrations of L*, a* and b* components. The L* and a* values were higher in the MAP than under VP systems while the b* values were higher in the VP samples than the MAP samples. Neither the bacterial cultures nor the packaging system influenced the textural property of ham. Starter cultures inoculated hams were rated superior (P < 0.05) in terms of their sensory properties. Hams packaged under MAP were rated superior (P < 0.05) than those packaged under VP in terms of appearance, colour, taste, tenderness, flavour, juiciness and overall acceptability.

A highly miniaturized vacuum package for a trapped ion atomic clock

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

Schwindt, Peter D. D., E-mail: pschwin@sandia.gov; Jau, Yuan-Yu; Partner, Heather

2016-05-15

We report on the development of a highly miniaturized vacuum package for use in an atomic clock utilizing trapped ytterbium-171 ions. The vacuum package is approximately 1 cm{sup 3} in size and contains a linear quadrupole RF Paul ion trap, miniature neutral Yb sources, and a non-evaporable getter pump. We describe the fabrication process for making the Yb sources and assembling the vacuum package. To prepare the vacuum package for ion trapping, it was evacuated, baked at a high temperature, and then back filled with a helium buffer gas. Once appropriate vacuum conditions were achieved in the package, it wasmore » sealed with a copper pinch-off and was subsequently pumped only by the non-evaporable getter. We demonstrated ion trapping in this vacuum package and the operation of an atomic clock, stabilizing a local oscillator to the 12.6 GHz hyperfine transition of {sup 171}Y b{sup +}. The fractional frequency stability of the clock was measured to be 2 × 10{sup −11}/τ{sup 1/2}.« less

Further characterisation of rotavirus cores: Ss(+)RNAs can be packaged in vitro but packaging lacks sequence specificity☆

PubMed Central

Desselberger, Ulrich; Richards, James; Tchertanov, Luba; Lepault, Jean; Lever, Andrew; Burrone, Oscar; Cohen, Jean

2013-01-01

Rotavirus (RV) cores were released from double-layered particles (DLPs) by high concentrations of CaCl2, purified and ‘opened’ by treatment with EDTA or EGTA. Under appropriate in vitro conditions DLPs have been shown to have transcriptase and ‘open cores’ replicase activity. Furthermore, it has been demonstrated that transcriptase activity and infectivity of native cores can be restored by transcapsidation with VP6, VP7 and VP4. The missing link for particle reconstitution in vitro has been the manipulation of ‘open cores’ to become functionally active cores again. The experiments described here were undertaken with the aim of exploring packaging of RV RNAs into opened cores in vitro. Rotavirus cores were opened by approximately 200 μM EGTA, leading to the release of genomic dsRNA. Conversely, RV cores were found to be stable in the presence of minimum concentrations of Ca2+, Mg2+, spermidine3+ and cobalthexamine3+ of between 40 and 300 μM. Aggregates of purified cores were resolved in the presence of 0.3 mM deoxycholate (minimum concentration). Core shells opened with EGTA were reconstituted by the addition of di- or trivalent cations within 2 min of the opening procedure. Addition of purified, baculovirus recombinant-expressed VP6 to native and reconstituted cores led to the formation of DLPs or DLP-like particles, which upon transfection into MA104 cells were infectious. The rescued infectivity likely originated in part from unopened and in part from reconstituted cores. Radiolabelled RV (+) ssRNAs could be packaged into reconstituted cores and DLPs, as indicated by resistance to RNase I digestion. The packaging reaction was, however, not RV RNA sequence-specific, since unrelated ssRNAs, such as those transcribed from HIV-2 cDNAs, were also packaged. The kinetics of packaging of homologous and heterologous RNAs were similar, as evidenced by competitive packaging assays. None of the packaged in vitro engineered RNA segments has so far been rescued into infectious virus. PMID:24091366

Effect of the ripening time under vacuum and packaging film permeability on processing and quality characteristics of low-fat fermented sausages.

PubMed

Liaros, N G; Katsanidis, E; Bloukas, J G

2009-12-01

The effect of vacuum ripening of low-fat fermented sausages packaged in films with different permeabilities on their microbiological, physicochemical and sensorial characteristics was studied. High-fat control sausages were produced with 30% initial fat and low-fat sausages with 10% initial fat. The low-fat sausages were separated into: (a) non-packaged (control) and (b) packaged under vacuum on 7th, 12th and 17th day of processing, remaining under vacuum during the ripening period for 21, 16 and 11days, respectively, in three different oxygen (100, 38 and⩽5cm(3)/m(2)/24h/1atm) and water vapour (4.5, <2.5 and 1g/m(2)24h) permeability plastic bags. Vacuum packaging reduced (p<0.05) the weight loss, the hardness and extent of lipid oxidation in the sausages, increased (p<0.05) their lightness, but had no effect (p>0.05) on the redness, compared to the control sausages. Packaging low-fat fermented sausages under vacuum for the last 11days of ripening in packaging film with high permeability increased (p<0.05) the lactic acid bacteria count. The same product packaged in film with medium permeability had a higher (p<0.05) Micrococcaceae count and the same (p>0.05) hardness and overall acceptability as the high-fat control sausages. A ripening time of 11days and the medium packaging film permeability were the most appropriate conditions for the vacuum packaging of low-fat fermented sausages.

Shelf life determination of sliced Portuguese traditional blood sausage--Morcela de Arroz de Monchique through microbiological challenge and consumer test.

PubMed

Pereira, Jorge A; Silva, Pedro; Matos, Teresa J S; Patarata, Luís

2015-03-01

Morcela de Arroz (MA) is a ready-to-eat blood and rice cooked sausage produced with pork, blood, rice, and seasonings, stuffed in natural casing and cooked above 90 °C/30 min. It is commercialized whole, not packed, with a restricted shelf life (1 wk/0 to 5 °C). The objective of this work was to establish sliced MA shelf life considering both the behavior of L. monocytogenes through a microbiological challenge test (MCT) and the consumer acceptability of MA stored: vacuum packed (VP), modified atmosphere packed (MAP: 80% CO2/20% N2 ), and aerobic packed (AP). The MCT was conducted inoculating ±3 log CFU/g of L. monocytogenes cell suspension on the MA slices. Packaged samples were stored at 3 ± 1 °C and 7 ± 1 °C until 20 d. At 3 ± 1 °C, L. monocytogenes behavior was not affected by packaging or storage time. At 7 ± 1 °C, the pathogen increased nearly 1 log CFU/g in the first 4 d. L. monocytogenes populations in AP were higher (P < 0.05) than in MAP. The pathogen may grow to hazardous levels in the 1st days if a temperature abuse occurs. Considering the acceptability by the consumers, the shelf life of MA stored at 3 ± 1 °C was 4.4 d for AP, 8.1 d for VP, and 10.4 d for MAP. The sensory shelf life established based on sensory spoilage is shorter than the shelf life to maintain the population of L. monocytogenes in safe levels. © 2015 Institute of Food Technologists®

Redesigning the continuous vacuum sealer packaging machine to improve the processing speed

NASA Astrophysics Data System (ADS)

Belo, J. B.; Widyanto, S. A.; Jamari, J.

2017-01-01

Vacuum sealer as a product packaging tool of food products to be able to vacuum air inside the plastic which is filled with food products and it causes the pressure lower. In this condition, the optimal heating temperature is reached in a shorter time, so that damage on plastic sealer of vacuumed food products could be prevented to be more effective and efficient. The purpose of this redesigning is to design a vacuum sealer packaging machine continuously through a conveyor mechanism on the packaging quality, time of processing speed of vacuuming food product in the plastic package. This designing process is conducted through several steps of designing and constructing tools until the products are ready to operate. Data analysis is done through quality test of vacuum and sealer to the plastic thickness of 75 µm, 80 µm, and 100 µm with temperature of 170°C, 180°C, 190°C and vacuum duration of 5 seconds, 8 seconds, and 60 seconds. Results of this designing process indicate that vacuum sealer works practically and more optimally with the time of vacuum processing speed of 0 to 1 minute/s; whereas, the pressure of vacuuming suction is until 1e-5 MPa. The results of tensile strength test are at a maximum of 32,796 (N/mm2) and a minimum of 20,155 (N/mm2) and the analysis of plastic composite with EDX. This result shows that the vacuum pressure and the quality of vacuum sealer are better and more efficient.

Study on vacuum packaging reliability of micromachined quartz tuning fork gyroscopes

NASA Astrophysics Data System (ADS)

Fan, Maoyan; Zhang, Lifang

2017-09-01

Packaging technology of the micromachined quartz tuning fork gyroscopes by vacuum welding has been experimentally studied. The performance of quartz tuning fork is influenced by the encapsulation shell, encapsulation method and fixation of forks. Alloy solder thick film is widely used in the package to avoid the damage of the chip structure by the heat resistance and hot temperature, and this can improve the device performance and welding reliability. The results show that the bases and the lids plated with gold and nickel can significantly improve the airtightness and reliability of the vacuum package. Vacuum packaging is an effective method to reduce the vibration damping, improve the quality factor and further enhance the performance. The threshold can be improved nearly by 10 times.

Quality of Meat (Longissimus dorsi) from Male Fallow Deer (Dama dama) Packaged and Stored under Vacuum and Modified Atmosphere Conditions

PubMed Central

Piaskowska, N.; Daszkiewicz, T.; Kubiak, D.; Zapotoczny, P.

2016-01-01

This study evaluated the effect of vacuum and modified atmosphere (40% CO2+60% N2, MA) packaging on the chemical composition, physicochemical properties and sensory attributes of chill-stored meat from 10 fallow deer (Dama dama) bucks at 17 to 18 months of age. The animals were hunter-harvested in the forests of north-eastern Poland. During carcass dressing (48 to 54 h post mortem), both musculus longissimus muscles were cut out. Each muscle was divided into seven sections which were allocated to three groups: 0, A, and B. Samples 0 were immediately subjected to laboratory analyses. Samples A were vacuum-packaged, and samples B were packaged in MA. Packaged samples were stored for 7, 14, and 21 days at 2°C. The results of the present study showed that the evaluated packaging systems had no significant effect on the quality of fallow deer meat during chilled storage. However, vacuum-packaged meat samples were characterised by greater drip loss. Vacuum and MA packaging contributed to preserving the desired physicochemical properties and sensory attributes of meat during 21 days of storage. Regardless of the packaging method used, undesirable changes in the colour, water-holding capacity and juiciness of meat, accompanied by tenderness improvement, were observed during chilled storage. PMID:27165026

Comparison of modified atmosphere packaging and vacuum packaging for long period storage of dry-cured ham: effects on colour, texture and microbiological quality.

PubMed

García-Esteban, Marta; Ansorena, Diana; Astiasarán, Iciar

2004-05-01

Slices of dry-cured hams (Biceps femoris muscle) were stored during 8 weeks under vacuum and modified atmospheres (100% N(2) and a mixture of 20% CO(2) and 80% N(2)) in order to study the modifications on colour, texture and microbial counts during that period. Lightness was found to be more stable when samples were stored with 20% CO(2) and 80% N(2) without statistical differences between vacuum and 100% N(2). A slight whiteness was observed in the vacuum packed samples. Yellowness increased during time in vacuum packed samples, although no differences were found among the three conditions at the end of the study. Redness values were not affected by time or by the packaging system. With regard to texture, values found for all samples were within the normal range for this type of products, although it was observed that modified atmosphere packaging preserved samples better from hardening than vacuum packaging. No safety problems were detected in relation to the microbial quality in any case. In general, no clear differences were found among the three packaging systems for colour, texture and microbial quality in the storage conditions studied.

Extension of the shelf life of prawns (Penaeus japonicus) by vacuum packaging and high-pressure treatment.

PubMed

López-Caballero, M E; Pérez-Mateos, M; Borderías, J A; Montero, P

2000-10-01

The present study has investigated the application of high pressures (200 and 400 MPa) in chilled prawn tails, both conventionally stored (air) and vacuum packaged. Vacuum packaging and high-pressure treatment did extend the shelf life of the prawn samples, although it did affect muscle color very slightly, giving it a whiter appearance. The viable shelf life of 1 week for the air-stored samples was extended to 21 days in the vacuum-packed samples, 28 days in the samples treated at 200 MPa, and 35 days in the samples pressurized at 400 MPa. Vacuum packaging checked the onset of blackening, whereas high-pressure treatment aggravated the problem. From a microbiological point of view, batches conventionally stored reached about 6 log CFU/g or even higher at 14 days. Similar figures were reached in total number of bacteria in vacuum-packed samples and in pressurized at 200-MPa samples at 21 days. When samples were pressurized at 400 MPa, total numbers of bacteria were below 5.5 log CFU/g at 35 days of storage. Consequently, a combination of vacuum packaging and high-pressure treatment would appear to be beneficial in prolonging freshness and preventing spotting.

An investigation of the effect of rapid slurry chilling on blown pack spoilage of vacuum-packaged beef primals.

PubMed

Reid, R; Fanning, S; Whyte, P; Kerry, J; Bolton, D

2017-02-01

The aim of this study was to investigate if rapid slurry chilling would retard or prevent blown pack spoilage (BPS) of vacuum-packaged beef primals. Beef primals were inoculated with Clostridium estertheticum subspp. estertheticum (DSMZ 8809), C. estertheticum subspp. laramenise (DSMZ 14864) and C. gasigenes (DSMZ 12272), and vacuum-packaged with and without heat shrinkage (90°C for 3 s). These packs were then subjected to immediate chilling in an ice slurry or using conventional blast chilling systems and stored at 2°C for up to 100 days. The onset and progress of BPS was monitored using the following scale; 0-no gas bubbles in drip; 1-gas bubbles in drip; 2-loss of vacuum; 3-'blown'; 4-presence of sufficient gas inside the packs to produce pack distension and 5-tightly stretched, 'overblown' packs/packs leaking. Rapid slurry chilling (as compared to conventional chilling) did not significantly affect (P > 0.05) the time to the onset or progress of BPS. It was therefore concluded that rapid chilling of vacuum-packaged beef primals, using an ice slurry system, may not be used as a control intervention to prevent or retard blown pack spoilage. This study adds to our growing understanding of blown pack spoilage of vacuum-packaged beef primals and suggests that rapid chilling of vacuum-packaged beef primals is not a control option for the beef industry. The results suggest that neither eliminating the heat shrinkage step nor rapid chilling of vacuum-packaged beef retard the time to blown pack spoilage. © 2016 The Society for Applied Microbiology.

Wafer-level vacuum/hermetic packaging technologies for MEMS

NASA Astrophysics Data System (ADS)

Lee, Sang-Hyun; Mitchell, Jay; Welch, Warren; Lee, Sangwoo; Najafi, Khalil

2010-02-01

An overview of wafer-level packaging technologies developed at the University of Michigan is presented. Two sets of packaging technologies are discussed: (i) a low temperature wafer-level packaging processes for vacuum/hermeticity sealing, and (ii) an environmentally resistant packaging (ERP) technology for thermal and mechanical control as well as vacuum packaging. The low temperature wafer-level encapsulation processes are implemented using solder bond rings which are first patterned on a cap wafer and then mated with a device wafer in order to encircle and encapsulate the device at temperatures ranging from 200 to 390 °C. Vacuum levels below 10 mTorr were achieved with yields in an optimized process of better than 90%. Pressures were monitored for more than 4 years yielding important information on reliability and process control. The ERP adopts an environment isolation platform in the packaging substrate. The isolation platform is designed to provide low power oven-control, vibration isolation and shock protection. It involves batch flip-chip assembly of a MEMS device onto the isolation platform wafer. The MEMS device and isolation structure are encapsulated at the wafer-level by another substrate with vertical feedthroughs for vacuum/hermetic sealing and electrical signal connections. This technology was developed for high performance gyroscopes, but can be applied to any type of MEMS device.

Structure of Ljungan virus provides insight into genome packaging of this picornavirus

NASA Astrophysics Data System (ADS)

Zhu, Ling; Wang, Xiangxi; Ren, Jingshan; Porta, Claudine; Wenham, Hannah; Ekström, Jens-Ola; Panjwani, Anusha; Knowles, Nick J.; Kotecha, Abhay; Siebert, C. Alistair; Lindberg, A. Michael; Fry, Elizabeth E.; Rao, Zihe; Tuthill, Tobias J.; Stuart, David I.

2015-10-01

Picornaviruses are responsible for a range of human and animal diseases, but how their RNA genome is packaged remains poorly understood. A particularly poorly studied group within this family are those that lack the internal coat protein, VP4. Here we report the atomic structure of one such virus, Ljungan virus, the type member of the genus Parechovirus B, which has been linked to diabetes and myocarditis in humans. The 3.78-Å resolution cryo-electron microscopy structure shows remarkable features, including an extended VP1 C terminus, forming a major protuberance on the outer surface of the virus, and a basic motif at the N terminus of VP3, binding to which orders some 12% of the viral genome. This apparently charge-driven RNA attachment suggests that this branch of the picornaviruses uses a different mechanism of genome encapsidation, perhaps explored early in the evolution of picornaviruses.

Structure of Ljungan virus provides insight into genome packaging of this picornavirus.

PubMed

Zhu, Ling; Wang, Xiangxi; Ren, Jingshan; Porta, Claudine; Wenham, Hannah; Ekström, Jens-Ola; Panjwani, Anusha; Knowles, Nick J; Kotecha, Abhay; Siebert, C Alistair; Lindberg, A Michael; Fry, Elizabeth E; Rao, Zihe; Tuthill, Tobias J; Stuart, David I

2015-10-08

Picornaviruses are responsible for a range of human and animal diseases, but how their RNA genome is packaged remains poorly understood. A particularly poorly studied group within this family are those that lack the internal coat protein, VP4. Here we report the atomic structure of one such virus, Ljungan virus, the type member of the genus Parechovirus B, which has been linked to diabetes and myocarditis in humans. The 3.78-Å resolution cryo-electron microscopy structure shows remarkable features, including an extended VP1 C terminus, forming a major protuberance on the outer surface of the virus, and a basic motif at the N terminus of VP3, binding to which orders some 12% of the viral genome. This apparently charge-driven RNA attachment suggests that this branch of the picornaviruses uses a different mechanism of genome encapsidation, perhaps explored early in the evolution of picornaviruses.

Lipid and protein oxidation and sensory properties of vacuum-packaged dry-cured ham subjected to high hydrostatic pressure.

PubMed

Fuentes, Verónica; Ventanas, Jesús; Morcuende, David; Estévez, Mario; Ventanas, Sonia

2010-07-01

The effect of HHP treatment (600 MPa) on the oxidative stability of lipids and proteins of vacuum-packaged Iberian dry-cured ham and the impact on the sensory characteristics of the product was investigated. In order to assess how different commercial presentations are affected by HHP treatment, three different presentations of vacuum-packaged Iberian dry-cured ham were considered, namely, (i) intact format (IF) corresponding to non-sliced vacuum-packaged dry-cured ham, (ii) conventional-sliced format (CSF) corresponding to dry-cured ham slices placed stretched out in the package and (iii) alternative-sliced format (ASF) corresponding to dry-cured ham slices piled up horizontally. The oxidation of dry-cured ham lipids and proteins was enhanced by HHP-treatment with the presentation being highly influential on these oxidative reactions. Pre-slicing dry-cured ham results in a more susceptible product to oxidative reactions during pressurisation and subsequent refrigerated storage. Possible mechanisms, by which HHP-induced oxidative reactions would affect particular sensory traits in vacuum-packaged Iberian dry-cured ham such as colour, texture and flavour attributes, are discussed. Copyright 2010 Elsevier Ltd. All rights reserved.

A conserved carboxy-terminal domain in the major tegument structural protein VP22 facilitates virion packaging of a chimeric protein during productive herpes simplex virus 1 infection

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

Schlegel, Elisabeth F.M.; Blaho, John A., E-mail: john.blaho@mssm.ed

2009-05-10

Recombinant virus HSV-1(RF177) was previously generated to examine tegument protein VP22 function by inserting the GFP gene into the gene encoding VP22. During a detailed analysis of this virus, we discovered that RF177 produces a novel fusion protein between the last 15 amino acids of VP22 and GFP, termed GCT-VP22. Thus, the VP22 carboxy-terminal specific antibody 22-3 and two anti-GFP antibodies reacted with an approximately 28 kDa protein from RF177-infected Vero cells. GCT-VP22 was detected at 1 and 3 hpi. Examination of purified virions indicated that GCT-VP22 was incorporated into RF177 virus particles. These observations imply that at least amore » portion of the information required for virion targeting is located in this domain of VP22. Indirect immunofluorescence analyses showed that GCT-VP22 also localized to areas of marginalized chromatin during RF177 infection. These results indicate that the last fifteen amino acids of VP22 participate in virion targeting during HSV-1 infection.« less

Effectiveness of some recent antimicrobial packaging concepts.

PubMed

Vermeiren, L; Devlieghere, F; Debevere, J

2002-01-01

A new type of active packaging is the combination of food-packaging materials with antimicrobial substances to control microbial surface contamination of foods. For both migrating and non-migrating antimicrobial materials, intensive contact between the food product and packaging material is required and therefore potential food applications include especially vacuum or skin-packaged products, e.g. vacuum-packaged meat, fish, poultry or cheese. Several antimicrobial compounds have been combined with different types of carriers (plastic and rubber articles, paper-based materials, textile fibrils and food-packaging materials). Until now, however, few antimicrobial concepts have found applications as a food-packaging material. Antimicrobial packaging materials cannot legally be used in the EU at the moment. The potential use would require amendments of several different legal texts involving areas such as food additives, food packaging, hygiene, etc. The main objective of this paper is to provide a state of the art about the different types of antimicrobial concepts, their experimental development and commercialization, and to present a case study summarizing the results of investigations on the feasibility of a low-density polyethylene (LDPE)-film containing triclosan to inhibit microbial growth on food surfaces and consequently prolong shelf-life or improve microbial food safety. In contrast with the strong antimicrobial effect in in-vitro simulated vacuum-packaged conditions against the psychrotrophic food pathogen L. monocytogenes, the 1000 mg kg(-1) containing triclosan film did not effectively reduce spoilage bacteria and growth of L. monocytogenes on refrigerated vacuum-packaged chicken breasts stored at 7 degrees C.

A wafer-level vacuum package using glass-reflowed silicon through-wafer interconnection for nano/micro devices.

PubMed

Jin, Joo-Young; Yoo, Seung-Hyun; Yoo, Byung-Wook; Kim, Yong-Kweon

2012-07-01

We propose a vacuum wafer-level packaging (WLP) process using glass-reflowed silicon via for nano/micro devices (NMDs). A through-wafer interconnection (TWIn) substrate with silicon vias and reflowed glass is introduced to accomplish a vertical feed-through of device. NMDs are fabricated in the single crystal silicon (SCS) layer which is formed on the TWIn substrate by Au eutectic bonding including Cr adhesion layer. The WLPof the devices is achieved with the capping glass wafer anodically bonded to the SCS layer. In order to demonstrate the successful hermetic packaging, we fabricated the micro-Pirani gauge in the SCS layer, and packaged it in the wafer-level. The vacuum level inside the packaging was measured to be 3.1 Torr with +/- 0.12 Torr uncertainty, and the packaging leakage was not detected during 24 hour after the packaging.

Effect of modified atmosphere and vacuum packaging on TVB-N production of rainbow trout (Oncorhynchus mykiss) and carp (Cyprinus carpio) cuts

NASA Astrophysics Data System (ADS)

Babić Milijašević, J.; Milijašević, M.; Đinović-Stojanović, J.; Vranić, D.

2017-09-01

The aim of our research was to examine the influence of packaging in modified atmosphere and vacuum on the total volatile basic nitrogen (TVB-N) content in muscle of rainbow trout (Oncorhynchus mykiss) and common carp (Cyprinus carpio), as well as to determine the most suitable gas mixtures for packing of these freshwater species. Three sample groups of trout and carp cuts were investigated. The two groups were packaged in modified atmosphere with different gas ratios: 90%CO2+10%N2 (MAP 1) and 60%CO2+40%N2 (MAP 2), whereas the third group of fish cuts were vacuum packaged. During trials, the trout and carp cuts were stored in refrigerator at 3°C±0.5°C. Determination of TVB-N was performed on 1, 4, 7, 9, 12 and 14 days of storage. The obtained results indicate that the investigated mixtures of gases and vacuum had a significant influence on the values of TVB-N in trout and carp cuts. The lowest increase in TVB-N was established in trout and carp cuts packaged in MAP 1, whereas the highest increase was established in vacuum packaged cuts. Based on the obtained results, it can be concluded that the gas mixture consisting of 90% CO2 and 10% N2 was the most suitable for packaging of fresh trout and carp cuts in terms of TVB-N value.

The effects of atmospheric pressure cold plasma treatment on microbiological, physical-chemical and sensory characteristics of vacuum packaged beef loin.

PubMed

Bauer, A; Ni, Y; Bauer, S; Paulsen, P; Modic, M; Walsh, J L; Smulders, F J M

2017-06-01

Effects on vacuum packaged and non-packaged beef longissimus samples exposed to atmospheric cold plasma (ACP) generated at different powers were studied over a 10day period of vacuum-, and a subsequent 3day period of aerobic storage. Exposure of non-covered beef samples under high power ACP conditions resulted in increased a*, b*, Chroma and Hue values, but ACP treatment of packaged loins did not impact colour (L*, a*, b*, Chroma, Hue), lipid peroxidation, sarcoplasmic protein denaturation, nitrate/nitrite uptake, or myoglobin isoform distribution. Colour values measured after 3days of aerobic storage following unpackaging (i.e. 20days post-mortem) were similar and all compliant with consumer acceptability standards. Exposure to ACP of the polyamide-polyethylene packaging film inoculated with Staphylococcus aureus, Listeria monocytogenes and two Escherichia coli strains resulted in >2 log reduction without affecting the integrity of the packaging matrix. Results indicate that ACP can reduce microbial numbers on surfaces of beef packages without affecting characteristics of the packaged beef. Copyright © 2017 Elsevier Ltd. All rights reserved.

Assessment of Performance of the Industrial Process of Bulk Vacuum Packaging of Raw Meat with Nondestructive Optical Oxygen Sensing Systems.

PubMed

Kelly, Caroline A; Cruz-Romero, Malco; Kerry, Joseph P; Papkovsky, Dmitri P

2018-05-02

The commercially-available optical oxygen-sensing system Optech-O₂ Platinum was applied to nondestructively assess the in situ performance of bulk, vacuum-packaged raw beef in three ~300 kg containers. Twenty sensors were attached to the inner surface of the standard bin-contained laminate bag (10 on the front and back sides), such that after filling with meat and sealing under vacuum, the sensors were accessible for optical interrogation with the external reader device. After filling and sealing each bag, the sensors were measured repetitively and nondestructively over a 15-day storage period at 1 °C, thus tracking residual oxygen distribution in the bag and changes during storage. The sensors revealed a number of unidentified meat quality and processing issues, and helped to improve the packaging process by pouring flakes of dry ice into the bag. Sensor utility in mapping the distribution of residual O₂ in sealed bulk containers and optimising and improving the packaging process, including handling and storage of bulk vacuum-packaged meat bins, was evident.

Safety analysis report for packaging (onsite) multicanister overpack cask

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

Edwards, W.S.

1997-07-14

This safety analysis report for packaging (SARP) documents the safety of shipments of irradiated fuel elements in the MUlticanister Overpack (MCO) and MCO Cask for a highway route controlled quantity, Type B fissile package. This SARP evaluates the package during transfers of (1) water-filled MCOs from the K Basins to the Cold Vacuum Drying Facility (CVDF) and (2) sealed and cold vacuum dried MCOs from the CVDF in the 100 K Area to the Canister Storage Building in the 200 East Area.

[Evaluation of the quality of poultry meat and its processing for vacuum packaging].

PubMed

Swiderski, F; Russel, S; Waszkiewicz-Robak, B; Cholewińska, E

1997-01-01

The aim of study was to evaluate the quality of poultry meat, roasted and smoked chicken and poultry pie packing under low and high vacuum. All investigated products were stored at +4 degrees C and evaluated by microbiological analysis. It was showed that packing under low and high vacuum inhibited development of aerobic microorganisms, proteolytic bacteria, yeasts and moulds. Vacuum-packaged storage of poultry meat and its products stimulated activity of anaerobic, nonsporeforming bacteria. The fast spoilage of fresh poultry meat was observed both under vacuum and conventional storage. The microbiology quality of poultry products depended on technology of production and microbiological quality of raw material.

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

Reynoso, F; Cho, S

Purpose: To develop and validate a Monte Carlo (MC) model of a Phillips RT-250 orthovoltage unit to test various beam spectrum modulation strategies for in vitro/vivo studies. A model of this type would enable the production of unconventional beams from a typical orthovoltage unit for novel therapeutic applications such as gold nanoparticle-aided radiotherapy. Methods: The MCNP5 code system was used to create a MC model of the head of RT-250 and a 30 × 30 × 30 cm{sup 3} water phantom. For the x-ray machine head, the current model includes the vacuum region, beryllium window, collimators, inherent filters and exteriormore » steel housing. For increased computational efficiency, the primary x-ray spectrum from the target was calculated from a well-validated analytical software package. Calculated percentage-depth-dose (PDD) values and photon spectra were validated against experimental data from film and Compton-scatter spectrum measurements. Results: The model was validated for three common settings of the machine namely, 250 kVp (0.25 mm Cu), 125 kVp (2 mm Al), and 75 kVp (2 mm Al). The MC results for the PDD curves were compared with film measurements and showed good agreement for all depths with a maximum difference of 4 % around dmax and under 2.5 % for all other depths. The primary photon spectra were also measured and compared with the MC results showing reasonable agreement between the two, validating the input spectra and the final spectra as predicted by the current MC model. Conclusion: The current MC model accurately predicted the dosimetric and spectral characteristics of each beam from the RT-250 orthovoltage unit, demonstrating its applicability and reliability for beam spectrum modulation tasks. It accomplished this without the need to model the bremsstrahlung xray production from the target, while significantly improved on computational efficiency by at least two orders of magnitude. Supported by DOD/PCRP grant W81XWH-12-1-0198.« less

Wafer-level hermetic vacuum packaging by bonding with a copper-tin thin film sealing ring

NASA Astrophysics Data System (ADS)

Akashi, Teruhisa; Funabashi, Hirofumi; Takagi, Hideki; Omura, Yoshiteru; Hata, Yoshiyuki

2018-04-01

A wafer-level hermetic vacuum packaging technology intended for use with MEMS devices was developed based on a copper-tin (CuSn) thin film sealing ring. To allow hermetic packaging, the shear strength of the CuSn thin film bond was improved by optimizing the pretreatment conditions. As a result, an average shear strength of 72.3 MPa was obtained and a cavity that had been hermetically sealed using wafer-level packaging (WLP) maintained its vacuum for 1.84 years. The total pressures in the cavities and the partial pressures of residual gases were directly determined with an ultra-low outgassing residual gas analyzer (RGA) system. Hermeticity was evaluated based on helium leak rates, which were calculated from helium pressures determined with the RGA system. The resulting data showed that a vacuum cavity following 1.84 years storage had a total pressure of 83.1 Pa, contained argon as the main residual gas and exhibited a helium leak rate as low as 1.67  ×  10-17 Pa · m3 s-1, corresponding to an air leak rate of 6.19  ×  10-18 Pa · m3 s-1. The RGA data demonstrate that WLP using a CuSn thin film sealing ring permits ultra-high hermeticity in conjunction with long-term vacuum packaging that is applicable to MEMS devices.

Effects of lactate and modified atmospheric packaging on premature browning in cooked ground beef patties.

PubMed

Mancini, R A; Ramanathan, R; Suman, S P; Konda, M K R; Joseph, P; Dady, G A; Naveena, B M; López-López, I

2010-06-01

Our objectives were to determine the effects of lactate and modified atmosphere packaging on raw surface color, lipid oxidation, and internal cooked color of ground beef patties. Eight chubs (85% lean) were divided in half and each half was either assigned to the control (no lactate) or mixed with 2.5% lactate (w/w). Following treatment, patties were prepared and packaged in either vacuum, PVC (atmospheric oxygen level), high-oxygen (80% O(2)+20% CO(2)), or 0.4% CO (30% CO(2)+69.6% N(2)) and stored for 0, 2, or 4days at 2 degrees C. After storage, raw surface color and lipid oxidation were measured and patties were cooked to either 66 degrees C or 71 degrees C. Lactate improved (p<0.05) color stability of PVC, high-oxygen, and vacuum packaged raw patties, but had no effect (p>0.05) on the a * values and visual color scores of patties in 0.4% CO. Lactate decreased (p<0.05) lipid oxidation in all packaging atmospheres. Nevertheless, high-oxygen and PVC-packaged patties had more (p<0.05) lipid oxidation than patties in CO and vacuum. Lactate had no effect (p>0.05) on premature browning, whereas patties packaged in high-oxygen demonstrated premature browning. Conversely, cooked patties in 0.4% CO and vacuum were more red (p<0.05) than both high-oxygen and PVC-packaged patties. Although lactate improved raw color stability, it did not minimize premature browning in cooked ground beef patties. Copyright 2010 Elsevier Ltd. All rights reserved.

A novel life cycle modeling system for Ebola virus shows a genome length-dependent role of VP24 in virus infectivity.

PubMed

Watt, Ari; Moukambi, Felicien; Banadyga, Logan; Groseth, Allison; Callison, Julie; Herwig, Astrid; Ebihara, Hideki; Feldmann, Heinz; Hoenen, Thomas

2014-09-01

Work with infectious Ebola viruses is restricted to biosafety level 4 (BSL4) laboratories, presenting a significant barrier for studying these viruses. Life cycle modeling systems, including minigenome systems and transcription- and replication-competent virus-like particle (trVLP) systems, allow modeling of the virus life cycle under BSL2 conditions; however, all current systems model only certain aspects of the virus life cycle, rely on plasmid-based viral protein expression, and have been used to model only single infectious cycles. We have developed a novel life cycle modeling system allowing continuous passaging of infectious trVLPs containing a tetracistronic minigenome that encodes a reporter and the viral proteins VP40, VP24, and GP1,2. This system is ideally suited for studying morphogenesis, budding, and entry, in addition to genome replication and transcription. Importantly, the specific infectivity of trVLPs in this system was ∼ 500-fold higher than that in previous systems. Using this system for functional studies of VP24, we showed that, contrary to previous reports, VP24 only very modestly inhibits genome replication and transcription when expressed in a regulated fashion, which we confirmed using infectious Ebola viruses. Interestingly, we also discovered a genome length-dependent effect of VP24 on particle infectivity, which was previously undetected due to the short length of monocistronic minigenomes and which is due at least partially to a previously unknown function of VP24 in RNA packaging. Based on our findings, we propose a model for the function of VP24 that reconciles all currently available data regarding the role of VP24 in nucleocapsid assembly as well as genome replication and transcription. Ebola viruses cause severe hemorrhagic fevers in humans, with no countermeasures currently being available, and must be studied in maximum-containment laboratories. Only a few of these laboratories exist worldwide, limiting our ability to study Ebola viruses and develop countermeasures. Here we report the development of a novel reverse genetics-based system that allows the study of Ebola viruses without maximum-containment laboratories. We used this system to investigate the Ebola virus protein VP24, showing that, contrary to previous reports, it only modestly inhibits virus genome replication and transcription but is important for packaging of genomes into virus particles, which constitutes a previously unknown function of VP24 and a potential antiviral target. We further propose a comprehensive model for the function of VP24 in nucleocapsid assembly. Importantly, on the basis of this approach, it should easily be possible to develop similar experimental systems for other viruses that are currently restricted to maximum-containment laboratories. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Quality attributes of farmed eel (Anguilla anguilla) stored under air, vacuum and modified atmosphere packaging at 0 degrees C.

PubMed

Arkoudelos, John; Stamatis, Nikolaos; Samaras, Fotis

2007-01-01

The shelf life of fresh eel in various packaging conditions of atmospheric air, vacuum and modified atmosphere packaging (MAP) (40% CO(2), 30% N(2) and 30% O(2)) at 0 degrees C was investigated. All raw eel samples received acceptable sensory scores during the first 11+/-1 days of storage in atmospheric air, 11+/-1 days of storage in vacuum and finally 18+/-1 days of storage in MAP conditions. Using the microbial quality indicators the shelf life of eel packed in air, vacuum and MAP was estimated to be more than 18, 28 and 34 days, respectively. The main spoilage microorganisms under MAP conditions were lactic acid producing bacteria followed by Shewanella spp., pseudomonads, Enterobacteriaceae and yeasts. Chemical data revealed that pH, ammonia, glucose and lactate examinations might not be useful for monitoring eel quality differences.

Effect of chemical sanitizer combined with modified atmosphere packaging on inhibiting Escherichia coli O157:H7 in commercial spinach.

PubMed

Lee, Sun-Young; Baek, Seung-Youb

2008-06-01

Escherichia coli O157:H7 contaminated spinach has recently caused several outbreaks of human illness in the USA and Canada. However, to date, there has been no study demonstrating an effective way to eliminate E. coli O157:H7 in spinach. Therefore, this study was conducted to investigate the effect of chemical sanitizers alone or in combination with packaging methods such as vacuum and modified atmosphere packaging (MAP) on inactivating E. coli O157:H7 in spinach during storage time. Spinach inoculated with E. coli O157:H7 was packaged in four different methods (air, vacuum, N(2) gas, and CO(2) gas packaging) following treatment with water, 100 ppm chlorine dioxide, or 100 ppm sodium hypochlorite for 5 min at room temperature and stored at 7+/-2 degrees C. Treatment with water did not significantly reduce levels of E. coli O157:H7 in spinach. However, treatment with chlorine dioxide and sodium hypochlorite significantly decreased levels of E. coli O157:H7 by 2.6 and 1.1 log(10)CFU/g, respectively. Levels of E. coli O157:H7 in samples packaged in air following treatments grew during storage time, whereas levels were maintained in samples packaged in other packaging methods (vacuum, N(2) gas, and CO(2) gas packaging). Therefore there were significant differences (about 3-4 log) of E. coli O157:H7 populations between samples packed in air and other packaging methods following treatment with chemical sanitizers after 7 days storage. These results suggest that the combination of treatment with chlorine dioxide and packaging methods such as vacuum and MAP may be useful for improving the microbial safety of spinach against E. coli O157:H7 during storage.

Effect of low-dose (1 kGy) gamma radiation and selected phosphates on the microflora of vacuum-packaged ground pork

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

Ehioba, R.M.

1987-01-01

The effects of low-dose (1 kGy) gamma radiation and selected phosphates on the microbiology of refrigerated, vacuum-packaged ground pork were studied. Low-dose gamma radiation reduced the numbers of naturally occurring mesophiles, psychrotrophs, and anaerobes. The effect of low-dose radiation on the populations of lactic acid bacteria was minimal. On storage of the irradiated vacuum-packaged ground pork at 5/sup 0/C, there was a partial bacterial recovery, suggesting sublethal bacterial injury due to irradiation. When 10/sup 7/ CFU/g of meat is taken to be the level beyond which the meat would be considered spoiled, uninoculated, vacuum-packaged ground pork treated with 1 kGymore » (100 krad) of gamma radiation had 3.5 more days of shelf-life in terms of psychrotrophic total counts. In relation to anaerobic bacterial numbers, meat shelf-life was extended 2.5 days, while the shelf-life of meat was extended 1 day in terms of aerobic mesophilic bacteria. Irradiation prolonged shelf-life in inoculated (10/sup 5/CFU/g) meat for 1.0-1.5 days. Addition of 0.4% sodium acid pyrophosphate (SAPP) contributed 2 additional days to inoculated, irradiated vacuum-packaged ground pork shelf-life. However, SAPP had no added effect on naturally occurring microflora. Irradiation greatly decreased the numbers of gram-negative microorganisms, resulting in predominance of the gram-positive, nonsporeforming Lactobacillus and coryneform bacteria.« less

Wafer-Level Vacuum Packaging of Smart Sensors.

PubMed

Hilton, Allan; Temple, Dorota S

2016-10-31

The reach and impact of the Internet of Things will depend on the availability of low-cost, smart sensors-"low cost" for ubiquitous presence, and "smart" for connectivity and autonomy. By using wafer-level processes not only for the smart sensor fabrication and integration, but also for packaging, we can further greatly reduce the cost of sensor components and systems as well as further decrease their size and weight. This paper reviews the state-of-the-art in the wafer-level vacuum packaging technology of smart sensors. We describe the processes needed to create the wafer-scale vacuum microchambers, focusing on approaches that involve metal seals and that are compatible with the thermal budget of complementary metal-oxide semiconductor (CMOS) integrated circuits. We review choices of seal materials and structures that are available to a device designer, and present techniques used for the fabrication of metal seals on device and window wafers. We also analyze the deposition and activation of thin film getters needed to maintain vacuum in the ultra-small chambers, and the wafer-to-wafer bonding processes that form the hermetic seal. We discuss inherent trade-offs and challenges of each seal material set and the corresponding bonding processes. Finally, we identify areas for further research that could help broaden implementations of the wafer-level vacuum packaging technology.

Wafer-Level Vacuum Packaging of Smart Sensors

PubMed Central

Hilton, Allan; Temple, Dorota S.

2016-01-01

The reach and impact of the Internet of Things will depend on the availability of low-cost, smart sensors—“low cost” for ubiquitous presence, and “smart” for connectivity and autonomy. By using wafer-level processes not only for the smart sensor fabrication and integration, but also for packaging, we can further greatly reduce the cost of sensor components and systems as well as further decrease their size and weight. This paper reviews the state-of-the-art in the wafer-level vacuum packaging technology of smart sensors. We describe the processes needed to create the wafer-scale vacuum microchambers, focusing on approaches that involve metal seals and that are compatible with the thermal budget of complementary metal-oxide semiconductor (CMOS) integrated circuits. We review choices of seal materials and structures that are available to a device designer, and present techniques used for the fabrication of metal seals on device and window wafers. We also analyze the deposition and activation of thin film getters needed to maintain vacuum in the ultra-small chambers, and the wafer-to-wafer bonding processes that form the hermetic seal. We discuss inherent trade-offs and challenges of each seal material set and the corresponding bonding processes. Finally, we identify areas for further research that could help broaden implementations of the wafer-level vacuum packaging technology. PMID:27809249

Ebola VP40 in Exosomes Can Cause Immune Cell Dysfunction

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

Pleet, Michelle L.; Mathiesen, Allison; DeMarino, Catherine

Ebola virus (EBOV) is an enveloped, ssRNA virus from the family Filoviridae capable of causing severe hemorrhagic fever with up to 80–90% mortality rates. The most recent outbreak of EBOV in West Africa starting in 2014 resulted in over 11,300 deaths; however, long-lasting persistence and recurrence in survivors has been documented, potentially leading to further transmission of the virus. We have previously shown that exosomes from cells infected with HIV-1, HTLV-1 and Rift Valley Fever virus are able to transfer viral proteins and non-coding RNAs to naïve recipient cells, resulting in an altered cellular activity. In the current manuscript, wemore » examined the effect of Ebola structural proteins VP40, GP, NP and VLPs on recipient immune cells, as well as the effect of exosomes containing these proteins on naïve immune cells. We found that VP40-transfected cells packaged VP40 into exosomes, and that these exosomes were capable of inducing apoptosis in recipient immune cells. In addition, we show that presence of VP40 within parental cells or in exosomes delivered to naïve cells could result in the regulation of RNAi machinery including Dicer, Drosha, and Ago 1, which may play a role in the induction of cell death in recipient immune cells. Exosome biogenesis was regulated by VP40 in transfected cells by increasing levels of ESCRT-II proteins EAP20 and EAP45, and exosomal marker proteins CD63 and Alix. VP40 was phosphorylated by Cdk2/Cyclin complexes at Serine 233 which could be reversed with r-Roscovitine treatment. The level of VP40-containing exosomes could also be regulated by treated cells with FDA-approved Oxytetracycline. Additionally, we utilized novel nanoparticles to safely capture VP40 and other viral proteins from Ebola VLPs spiked into human samples using SDS/reducing agents, thus minimizing the need for BSL-4 conditions for most downstream assays. Collectively, our data indicates that VP40 packaged into exosomes may be responsible for the deregulation and eventual destruction of the T-cell and myeloid arms of the immune system (bystander lymphocyte apoptosis), allowing the virus to replicate to high titers in the immunocompromised host. Moreover, our results suggest that the use of drugs such as Oxytetracycline to modulate the levels of exosomes exiting EBOV-infected cells may be able to prevent the devastation of the adaptive immune system and allow for an improved rate of survival.« less

Ebola VP40 in Exosomes Can Cause Immune Cell Dysfunction

DOE PAGES

Pleet, Michelle L.; Mathiesen, Allison; DeMarino, Catherine; ...

2016-11-07

Ebola virus (EBOV) is an enveloped, ssRNA virus from the family Filoviridae capable of causing severe hemorrhagic fever with up to 80–90% mortality rates. The most recent outbreak of EBOV in West Africa starting in 2014 resulted in over 11,300 deaths; however, long-lasting persistence and recurrence in survivors has been documented, potentially leading to further transmission of the virus. We have previously shown that exosomes from cells infected with HIV-1, HTLV-1 and Rift Valley Fever virus are able to transfer viral proteins and non-coding RNAs to naïve recipient cells, resulting in an altered cellular activity. In the current manuscript, wemore » examined the effect of Ebola structural proteins VP40, GP, NP and VLPs on recipient immune cells, as well as the effect of exosomes containing these proteins on naïve immune cells. We found that VP40-transfected cells packaged VP40 into exosomes, and that these exosomes were capable of inducing apoptosis in recipient immune cells. In addition, we show that presence of VP40 within parental cells or in exosomes delivered to naïve cells could result in the regulation of RNAi machinery including Dicer, Drosha, and Ago 1, which may play a role in the induction of cell death in recipient immune cells. Exosome biogenesis was regulated by VP40 in transfected cells by increasing levels of ESCRT-II proteins EAP20 and EAP45, and exosomal marker proteins CD63 and Alix. VP40 was phosphorylated by Cdk2/Cyclin complexes at Serine 233 which could be reversed with r-Roscovitine treatment. The level of VP40-containing exosomes could also be regulated by treated cells with FDA-approved Oxytetracycline. Additionally, we utilized novel nanoparticles to safely capture VP40 and other viral proteins from Ebola VLPs spiked into human samples using SDS/reducing agents, thus minimizing the need for BSL-4 conditions for most downstream assays. Collectively, our data indicates that VP40 packaged into exosomes may be responsible for the deregulation and eventual destruction of the T-cell and myeloid arms of the immune system (bystander lymphocyte apoptosis), allowing the virus to replicate to high titers in the immunocompromised host. Moreover, our results suggest that the use of drugs such as Oxytetracycline to modulate the levels of exosomes exiting EBOV-infected cells may be able to prevent the devastation of the adaptive immune system and allow for an improved rate of survival.« less

Ebola VP40 in Exosomes Can Cause Immune Cell Dysfunction.

PubMed

Pleet, Michelle L; Mathiesen, Allison; DeMarino, Catherine; Akpamagbo, Yao A; Barclay, Robert A; Schwab, Angela; Iordanskiy, Sergey; Sampey, Gavin C; Lepene, Benjamin; Nekhai, Sergei; Aman, M J; Kashanchi, Fatah

2016-01-01

Ebola virus (EBOV) is an enveloped, ssRNA virus from the family Filoviridae capable of causing severe hemorrhagic fever with up to 80-90% mortality rates. The most recent outbreak of EBOV in West Africa starting in 2014 resulted in over 11,300 deaths; however, long-lasting persistence and recurrence in survivors has been documented, potentially leading to further transmission of the virus. We have previously shown that exosomes from cells infected with HIV-1, HTLV-1 and Rift Valley Fever virus are able to transfer viral proteins and non-coding RNAs to naïve recipient cells, resulting in an altered cellular activity. In the current manuscript, we examined the effect of Ebola structural proteins VP40, GP, NP and VLPs on recipient immune cells, as well as the effect of exosomes containing these proteins on naïve immune cells. We found that VP40-transfected cells packaged VP40 into exosomes, and that these exosomes were capable of inducing apoptosis in recipient immune cells. Additionally, we show that presence of VP40 within parental cells or in exosomes delivered to naïve cells could result in the regulation of RNAi machinery including Dicer, Drosha, and Ago 1, which may play a role in the induction of cell death in recipient immune cells. Exosome biogenesis was regulated by VP40 in transfected cells by increasing levels of ESCRT-II proteins EAP20 and EAP45, and exosomal marker proteins CD63 and Alix. VP40 was phosphorylated by Cdk2/Cyclin complexes at Serine 233 which could be reversed with r-Roscovitine treatment. The level of VP40-containing exosomes could also be regulated by treated cells with FDA-approved Oxytetracycline. Additionally, we utilized novel nanoparticles to safely capture VP40 and other viral proteins from Ebola VLPs spiked into human samples using SDS/reducing agents, thus minimizing the need for BSL-4 conditions for most downstream assays. Collectively, our data indicates that VP40 packaged into exosomes may be responsible for the deregulation and eventual destruction of the T-cell and myeloid arms of the immune system (bystander lymphocyte apoptosis), allowing the virus to replicate to high titers in the immunocompromised host. Moreover, our results suggest that the use of drugs such as Oxytetracycline to modulate the levels of exosomes exiting EBOV-infected cells may be able to prevent the devastation of the adaptive immune system and allow for an improved rate of survival.

Structure and dynamics of Ebola virus matrix protein VP40 by a coarse-grained Monte Carlo simulation

NASA Astrophysics Data System (ADS)

Pandey, Ras; Farmer, Barry

Ebola virus matrix protein VP40 (consisting of 326 residues) plays a critical role in viral assembly and its functions such as regulation of viral transcription, packaging, and budding of mature virions into the plasma membrane of infected cells. How does the protein VP40 go through structural evolution during the viral life cycle remains an open question? Using a coarse-grained Monte Carlo simulation we investigate the structural evolution of VP40 as a function of temperature with the input of a knowledge-based residue-residue interaction. A number local and global physical quantities (e.g. mobility profile, contact map, radius of gyration, structure factor) are analyzed with our large-scale simulations. Our preliminary data show that the structure of the protein evolves through different state with well-defined morphologies which can be identified and quantified via a detailed analysis of structure factor.

Shelf life of cooked goat blood sausage prepared with the addition of heart and kidney.

PubMed

Silva, F A P; Amaral, D S; Guerra, I C D; Arcanjo, N M O; Bezerra, T K A; Ferreira, V C S; Araújo, I B S; Dalmás, P S; Madruga, M S

2014-08-01

This study focused on the effect of two packaging formats (vacuum packaging and over-wrap packaging) on the shelf life of cooked sausage prepared with blood, heart, kidney and goat meat fragments under storage at 4±1°C for a period of 90 days. The storage time and type of packaging significantly affected the chemical (pH, moisture, protein and TBARS number), physical (shear force) and microbial (mould and yeast) parameters of cooked sausage. Vacuum packaging maintained the microbiological and chemical qualities of cooked goat blood sausage for a longer period of time (63 days) compared to over-wrap packaging (41 days) and could be a viable alternative to refrigerated storage of the product for quality maintenance. Copyright © 2014 Elsevier Ltd. All rights reserved.

[Assessment of the probability of encountering staphylococcal enterotoxins in lactic acid cheese packaged in laminates].

PubMed

Steinka, Izabela

2004-01-01

Immunoassay methods were used to identify the presence of staphylococcal enterotoxins in lactic acid cheese vacuum and non-vacuum packed. There was assessed the probability of encountering staphylococcal enterotoxin in cheese dependent on different systems of packaging, count of staphylococcal cells, intensiveness of coagulase synthesis and tightness of packaging. The presence of enterotoxin was identified in 5% of researched samples of products stored for 14 days. The influence of packaging system and tightness on presence of enterotoxin was observed. The probability of presence of staphylococcal and enterotoxin in relation to researched factors was presented by the mathematical models.

Effect of modified atmosphere and vacuum packaging on some quality characteristics and the shelf-life of "morcilla", a typical cooked blood sausage.

PubMed

Cachaldora, Aida; García, Gloria; Lorenzo, José M; García-Fontán, M Camino

2013-02-01

The effect of modified atmosphere and vacuum packaging on the shelf-life of "morcilla", a traditional cooked blood sausage, was investigated. A total of 99 "morcillas" were packaged under vacuum and in modified atmosphere using three different gas mixtures: 15:35:50/O(2):N(2):CO(2) (atmosphere 1), 60:40/N(2):CO(2) (atmosphere 2) and 40:60/N(2):CO(2) (atmosphere 3), and stored during 2, 4, 6 and 8 weeks at 4 °C. Shelf life evaluation was based on pH, water activity (a(w)), colour (CIE L*, a*, b*, C* and h*), TBARS formation and microbial counts. The results indicated that, in general, storage time affected (P<0.05) all parameters whereas no significant differences were observed (P>0.05) among packaging conditions. Based on the microbial counts, the shelf-life of "morcilla" would be greater than 8 weeks for all packaging conditions. Samples packaged with high CO(2) concentrations (40:60/N(2):CO(2)) showed the lowest values of TBARS at the end of storage. Copyright © 2012 Elsevier Ltd. All rights reserved.

Effect of a nano-silver coating on the quality of fresh turkey meat during storage after modified atmosphere or vacuum packaging.

PubMed

Deus, D; Kehrenberg, C; Schaudien, D; Klein, G; Krischek, C

2017-02-01

Nano-silver is used in consumer products due to its antibacterial properties. The aim of this study was to evaluate the effect of a nano-silver-coated film on the quality of turkey meat during vacuum-sealed and modified atmosphere packaging up to 12 days of storage. In the first part of the experiment, turkey breasts were packaged using either vacuum packaging or modified atmosphere packages (MAPs) and contained films with or without a nano-silver coating (control film). Parameters such as pH, electrical conductivity, color (lightness L*, redness a*), myoglobin redox forms, thiobarbituric acid-reactive substances (TBARS), biogenic amines (BAs), total viable bacterial counts, Pseudomonas species counts, and Enterobacteriaceae species counts were evaluated on storage days 4, 8, and 12. In the second part of the study, the antimicrobial effect of a nano-silver-coated film on turkey breast was evaluated after inoculation with Escherichia coli (E. coli). Turkey meat packaged with the nano-silver film exhibited lower a* values on days 1 (3.15 ± 0.62), 4 (3.90 ± 0.68), and 8 (4.27 ± 0.76) compared to the packaged meat with the control film (3.41 ± 0.73, 4.35 ± 0.94, 4.85 ± 0.89, respectively), indicating special optical properties of nanoparticles. Concerning the BAs, silver packaged meat showed higher values of tyramine on day 12 (1274 ± 392 ng/g meat) and cadaverine on day 4 (1224 ± 435 ng/g meat) compared to the normal packaged products (647 ± 576 and 508 ± 314 ng/g meat, respectively). MAP meat revealed higher L* and TBARS values and lower microbial counts than the vacuum packaged products on all days. The MAP meat also showed lower a* results on days 4 and 8 and higher metmyoglobin (metMb) values on days 8 and 12 compared to th E: vacuum products. In the inoculation study, the microbial counts of the turkey meat were comparable between the two film types. The study showed that the nano-silver coating did not exhibit any advantageous effects on the quality and microbiological parameters of the turkey meat. © 2016 Poultry Science Association Inc.

Quality of hurdle treated pork sausages during refrigerated (4 ± 1°C) storage.

PubMed

Thomas, R; Anjaneyulu, A S R; Kondaiah, N

2010-06-01

Pork sausages developed using hurdle technology was evaluated during refrigerated storage (4 ± 1°C). Hurdles incorporated were low pH, low water activity, vacuum packaging and post package reheating. Dipping in potassium sorbate solution prior to vacuum packaging was also tried. Hurdle treatment significantly (p <0.05) reduced the rate of deterioration of quality characteristics of pork sausages during storage, as indicated by TBARS and tyrosine values. Incorporation of hurdles decreased the growth of different spoilage and pathogenic microorganisms. Combination of pH, water activity, vacuum packaging and reheating inhibited the growth of yeast and molds up to 12 days, while additional dipping of sausages in 1% potassium sorbate solution prior to packaging inhibited their growth even on 30(th) day of storage. Incorporation of hurdles resulted in initial reduction in all the sensory attributes, but they helped to maintain these attributes for significantly longer period compared to control. Hurdle treated sausages exhibited no spoilage signs even on day 30, while the control sausages were found acceptable only up to 18 days.

Surface attachment of active antimicrobial coatings onto conventional plastic-based laminates and performance assessment of these materials on the storage life of vacuum packaged beef sub-primals.

PubMed

Clarke, David; Tyuftin, Andrey A; Cruz-Romero, Malco C; Bolton, Declan; Fanning, Seamus; Pankaj, Shashi K; Bueno-Ferrer, Carmen; Cullen, Patrick J; Kerry, Joe P

2017-04-01

Two antimicrobial coatings, namely Sodium octanoate and Auranta FV (a commercial antimicrobial composed of bioflavonoids, citric, malic, lactic, and caprylic acids) were used. These two antimicrobials were surface coated onto the inner polyethylene layer of cold plasma treated polyamide films using beef gelatin as a carrier and coating polymer. This packaging material was then used to vacuum pack beef sub-primal cuts and stored at 4 °C. A control was prepared using the non-coated commercial laminate and the same vacuum packaged sub-primal beef cuts. During storage, microbial and quality assessments were carried out. Sodium octanoate treated packages significantly (p < 0.05) reduced microbial counts for all bacteria tested with an increase of 7 and 14 days, respectively compared to control samples. No significant effect on pH was observed with any treatment. The results suggested that these food grade antimicrobials have the potential to be used in antimicrobial active packaging applications for beef products. Copyright © 2016 Elsevier Ltd. All rights reserved.

Combined effects of gamma irradiation and a modified atmospheric packaging on the physicochemical characteristics of sausage

NASA Astrophysics Data System (ADS)

Ahn, Hyun-Joo; Kim, Jae-Hyun; Jo, Cheorun; Lee, Ju-Woon; Yook, Hong-Sun; Kim, Hee-Yun; Byun, Myung-Woo

2004-09-01

This study is to investigate the combined effects of irradiation and a modified atmospheric packaging (MAP) on the color, nitrosoheme pigments (NO-Mb), residual nitrite and N-nitrosodimethylamine (NDMA) in sausage during storage. Sausage with air, vacuum, CO 2, N 2, or CO 2/N 2 packaging was irradiated at 5 kGy. Irradiation reduced the red color of sausage, and a vacuum or MAP was effective in minimizing the loss of redness. The reduction of NO-Mb was observed by irradiation, while the MAP was more effective in maintaining the NO-Mb than the aerobic ones. Residual nitrite was reduced by irradiation, and the contents were lower under vacuum or MAP than aerobic ones. NDMA was significantly reduced by irradiation.

Effect of different packaging methods and storage temperature on microbiological and physicochemical quality characteristics of meatball.

PubMed

Yilmaz, I; Demirci, M

2010-06-01

The objective of this research was to determine physicochemical changes and microbiological quality of the different packaged meatball samples. Meatball samples in polystyrene tray were closed with polyethylene film (PS packs), vacuumed and modified atmosphere packaged, (MAP) (65% N(2), 35% CO(2)), and held under refrigerated display (4 °C) for 8, 16 and 16 days for PS packs, vacuum and MAP, respectively. Microbial load, free fatty acids and thiobarbituric acid values of the samples tended to increase with storage time. Bacteria counts of the raw meatball samples increased 2 log cycles at the end of storage compared with initial values. Meatball samples can be stored without any microbiological problem for 7 days at 4 °C. Results from this study suggested that shelf-life assigned to modified-MAP and vacuum-packed meatballs may be appropriate. Meatball samples underwent physical deformation when they were packed before vacuum process. With these negative factors considered, MAP is superior to other two packs methods.

Relevance of nanocomposite packaging on the stability of vacuum-packed dry cured ham.

PubMed

Lloret, Elsa; Fernandez, Avelina; Trbojevich, Raul; Arnau, Jacint; Picouet, Pierre A

2016-08-01

In this study effects of a novel high barrier multilayer polyamide film containing dispersed nanoclays (PAN) on the stability of vacuum packed dry-cured ham were investigated during 90days refrigerated storage in comparison with non-modified multilayer polyamide (PA) and a commercial high barrier film. Characteristic bands of the mineral in FT-IR spectra confirmed the presence of nanoclays in PAN, enhancing oxygen transmission barrier properties and UV protection. Packaging in PAN films did not originate significant changes on colour or lipid oxidation during prolonged storage of vacuum-packed dry-cured ham. Larger oxygen transmission rates in PA films caused changes in CIE b* during refrigerated storage. Ham quality was not affected by light exposition during 90days and only curing had a significant benefit on colour and TBARS, being cured samples more stable during storage in all the packages used. Packaging of dry-cured ham in PAN was equivalent to commercial high barrier films. Copyright © 2016 Elsevier Ltd. All rights reserved.

Structural dissection of Ebola virus and its assembly determinants using cryo-electron tomography.

PubMed

Bharat, Tanmay A M; Noda, Takeshi; Riches, James D; Kraehling, Verena; Kolesnikova, Larissa; Becker, Stephan; Kawaoka, Yoshihiro; Briggs, John A G

2012-03-13

Ebola virus is a highly pathogenic filovirus causing severe hemorrhagic fever with high mortality rates. It assembles heterogenous, filamentous, enveloped virus particles containing a negative-sense, single-stranded RNA genome packaged within a helical nucleocapsid (NC). We have used cryo-electron microscopy and tomography to visualize Ebola virus particles, as well as Ebola virus-like particles, in three dimensions in a near-native state. The NC within the virion forms a left-handed helix with an inner nucleoprotein layer decorated with protruding arms composed of VP24 and VP35. A comparison with the closely related Marburg virus shows that the N-terminal region of nucleoprotein defines the inner diameter of the Ebola virus NC, whereas the RNA genome defines its length. Binding of the nucleoprotein to RNA can assemble a loosely coiled NC-like structure; the loose coil can be condensed by binding of the viral matrix protein VP40 to the C terminus of the nucleoprotein, and rigidified by binding of VP24 and VP35 to alternate copies of the nucleoprotein. Four proteins (NP, VP24, VP35, and VP40) are necessary and sufficient to mediate assembly of an NC with structure, symmetry, variability, and flexibility indistinguishable from that in Ebola virus particles released from infected cells. Together these data provide a structural and architectural description of Ebola virus and define the roles of viral proteins in its structure and assembly.

Joint inversion for Vp, Vs, and Vp/Vs at SAFOD, Parkfield, California

USGS Publications Warehouse

Zhang, H.; Thurber, C.; Bedrosian, P.

2009-01-01

We refined the three-dimensional (3-D) Vp, Vs and Vp/Vs models around the San Andreas Fault Observatory at Depth (SAFOD) site using a new double-difference (DD) seismic tomography code (tomoDDPS) that simultaneously solves for earthquake locations and all three velocity models using both absolute and differential P, S, and S-P times. This new method is able to provide a more robust Vp/Vs model than that from the original DD tomography code (tomoDD), obtained simply by dividing Vp by Vs. For the new inversion, waveform cross-correlation times for earthquakes from 2001 to 2002 were also used, in addition to arrival times from earthquakes and explosions in the region. The Vp values extracted from the model along the SAFOD trajectory match well with the borehole log data, providing in situ confirmation of our results. Similar to previous tomographic studies, the 3-D structure around Parkfield is dominated by the velocity contrast across the San Andreas Fault (SAF). In both the Vp and Vs models, there is a clear low-velocity zone as deep as 7 km along the SAF trace, compatible with the findings from fault zone guided waves. There is a high Vp/Vs anomaly zone on the southwest side of the SAF trace that is about 1-2 km wide and extends as deep as 4 km, which is interpreted to be due to fluids and fractures in the package of sedimentary rocks abutting the Salinian basement rock to the southwest. The relocated earthquakes align beneath the northeast edge of this high Vp/Vs zone. We carried out a 2-D correlation analysis for an existing resistivity model and the corresponding profiles through our model, yielding a classification that distinguishes several major lithologies. ?? 2009 by the American Geophysical Union.

Vacuum decay container/closure integrity testing technology. Part 2. Comparison to dye ingress tests.

PubMed

Wolf, Heinz; Stauffer, Tony; Chen, Shu-Chen Y; Lee, Yoojin; Forster, Ronald; Ludzinski, Miron; Kamat, Madhav; Mulhall, Brian; Guazzo, Dana Morton

2009-01-01

Part 1 of this series demonstrated that a container closure integrity test performed according to ASTM F2338-09 Standard Test Method for Nondestructive Detection of Leaks in Packages by Vacuum Decay Method using a VeriPac 325/LV vacuum decay leak tester by Packaging Technologies & Inspection, LLC (PTI) is capable of detecting leaks > or = 5.0 microm (nominal diameter) in rigid, nonporous package systems, such as prefilled glass syringes. The current study compared USP, Ph.Eur. and ISO dye ingress integrity test methods to PTI's vacuum decay technology for the detection of these same 5-, 10-, and 15-microm laser-drilled hole defects in 1-mL glass prefilled syringes. The study was performed at three test sites using several inspectors and a variety of inspection conditions. No standard dye ingress method was found to reliably identify all holed syringes. Modifications to these standard dye tests' challenge conditions increased the potential for dye ingress, and adjustments to the visual inspection environment improved dye ingress detection. However, the risk of false positive test results with dye ingress tests remained. In contrast, the nondestructive vacuum decay leak test method reliably identified syringes with holes > or = 5.0 microm.

Localized heating and bonding technique for MEMS packaging

NASA Astrophysics Data System (ADS)

Cheng, Yu-Ting

Localized heating and bonding techniques have been developed for hermetic and vacuum packaging of MEMS devices, including silicon-to-glass fusion, silicon-gold eutectic, and silicon-to-glass bonding using PSG, indium, aluminum, and aluminum/silicon alloy as the intermediate layer. Line shaped phosphorus-doped polysilicon or gold films are used as resistive microheaters to provide enough thermal energy for bonding. The bonding processes are conducted in the common environment of room temperature and atmospheric pressure and can achieve bonding strength comparable to the fracture toughness of bulk silicon in less than 10 minutes. About 5 watts of input power is needed for localized bonding which can seal a 500 x 500 mum2 area. The total input power is determined by the thermal properties of bonding materials, including the heat capacity and latent heat. Two important bonding results are obtained: (1) The surface step created by the electrical interconnect line can be planarized by reflowing the metal solder. (2) Small applied pressure, less than 1MPa, for intimate contact reduces mechanical damage to the device substrate. This new class of bonding technology has potential applications for MEMS fabrication and packaging that require low temperature processing at the wafer level, excellent bonding strength and hermetic sealing characteristics. A hermetic package based on localized aluminum/silicon-to-glass bonding has been successfully fabricated. Less than 0.2 MPa contact pressure with 46mA input current for two parallel 3.5mum wide polysilicon on-chip microheaters can create as high as 700°C bonding temperature and achieve a strong and reliable bond in 7.5 minutes. Accelerated testing in an autoclave shows some packages survive more than 450 hours under 3 atm, 100%RH and 128°C. Premature failure has been attributed to some unbonded regions on the failed samples. The bonding yield and reliability have been improved by increasing bonding time and applied pressure. Finally, vacuum encapsulation of folded-beam comb-drive mu-resonators used as pressure monitors has been demonstrated using localized aluminum/silicon-to-glass bonding. With 3.4 watt heating power, ˜0.2MPa applied contact pressure, and 90 minutes wait time before bonding, vacuum encapsulation can be achieved with the same vacuum level as the packaging environment which is about 25 mtorr. Metal coating used as diffusion barrier and a longer wait time before bonding are used to improve the vacuum level of the package. Long-term measurement of the Q of un-annealed vacuum-packaged mu-resonators, illustrates stable operation after 19 weeks.

How to Build a Vacuum Spring-transport Package for Spinning Rotor Gauges

PubMed Central

Fedchak, James A.; Scherschligt, Julia; Sefa, Makfir

2016-01-01

The spinning rotor gauge (SRG) is a high-vacuum gauge often used as a secondary or transfer standard for vacuum pressures in the range of 1.0 x 10-4 Pa to 1.0 Pa. In this application, the SRGs are frequently transported to laboratories for calibration. Events can occur during transportation that change the rotor surface conditions, thus changing the calibration factor. To assure calibration stability, a spring-transport mechanism is often used to immobilize the rotor and keep it under vacuum during transport. It is also important to transport the spring-transport mechanism using packaging designed to minimize the risk of damage during shipping. In this manuscript, a detailed description is given on how to build a robust spring-transport mechanism and shipping container. Together these form a spring-transport package. The spring-transport package design was tested using drop-tests and the performance was found to be excellent. The present spring-transport mechanism design keeps the rotor immobilized when experiencing shocks of several hundred g (g = 9.8 m/sec2 and is the acceleration due to gravity), while the shipping container assures that the mechanism will not experience shocks greater than about 100 g during common shipping mishaps (as defined by industry standards). PMID:27078575

Vacuum decay container closure integrity leak test method development and validation for a lyophilized product-package system.

PubMed

Patel, Jayshree; Mulhall, Brian; Wolf, Heinz; Klohr, Steven; Guazzo, Dana Morton

2011-01-01

A leak test performed according to ASTM F2338-09 Standard Test Method for Nondestructive Detection of Leaks in Packages by Vacuum Decay Method was developed and validated for container-closure integrity verification of a lyophilized product in a parenteral vial package system. This nondestructive leak test method is intended for use in manufacturing as an in-process package integrity check, and for testing product stored on stability in lieu of sterility tests. Method development and optimization challenge studies incorporated artificially defective packages representing a range of glass vial wall and sealing surface defects, as well as various elastomeric stopper defects. Method validation required 3 days of random-order replicate testing of a test sample population of negative-control, no-defect packages and positive-control, with-defect packages. Positive-control packages were prepared using vials each with a single hole laser-drilled through the glass vial wall. Hole creation and hole size certification was performed by Lenox Laser. Validation study results successfully demonstrated the vacuum decay leak test method's ability to accurately and reliably detect those packages with laser-drilled holes greater than or equal to approximately 5 μm in nominal diameter. All development and validation studies were performed at Whitehouse Analytical Laboratories in Whitehouse, NJ, under the direction of consultant Dana Guazzo of RxPax, LLC, using a VeriPac 455 Micro Leak Test System by Packaging Technologies & Inspection (Tuckahoe, NY). Bristol Myers Squibb (New Brunswick, NJ) fully subsidized all work. A leak test performed according to ASTM F2338-09 Standard Test Method for Nondestructive Detection of Leaks in Packages by Vacuum Decay Method was developed and validated to detect defects in stoppered vial packages containing lyophilized product for injection. This nondestructive leak test method is intended for use in manufacturing as an in-process package integrity check, and for testing product stored on stability in lieu of sterility tests. Test method validation study results proved the method capable of detecting holes laser-drilled through the glass vial wall greater than or equal to 5 μm in nominal diameter. Total test time is less than 1 min per package. All method development and validation studies were performed at Whitehouse Analytical Laboratories in Whitehouse, NJ, under the direction of consultant Dana Guazzo of RxPax, LLC, using a VeriPac 455 Micro Leak Test System by Packaging Technologies & Inspection (Tuckahoe, NY). Bristol Myers Squibb (New Brunswick, NJ) fully subsidized all work.

Nitrogen reactive ion etch processes for the selective removal of poly-(4-vinylpyridine) in block copolymer films.

PubMed

Flynn, Shauna P; Bogan, Justin; Lundy, Ross; Khalafalla, Khalafalla E; Shaw, Matthew; Rodriguez, Brian J; Swift, Paul; Daniels, Stephen; O'Connor, Robert; Hughes, Greg; Kelleher, Susan M

2018-08-31

Self-assembling block copolymer (BCP) patterns are one of the main contenders for the fabrication of nanopattern templates in next generation lithography technology. Transforming these templates to hard mark materials is key for pattern transfer and in some cases, involves selectively removing one block from the nanopattern. For poly(styrene)-block-poly(4-vinylpyridine) (PS-b-P4VP), a high χ BCP system which could be potentially incorporated into semiconductor nanofabrication, this selective removal is predominantly done by a wet etch/activation process. Conversely, this process has numerous disadvantages including lack of control and high generation of waste leading to high cost. For these reasons, our motivation was to move away from the wet etch process and optimise a dry etch which would overcome the limitations associated with the activation process. The work presented herein shows the development of a selective plasma etch process for the removal of P4VP cores from PS-b-P4VP nanopatterned film. Results have shown that a nitrogen reactive ion etch plasma has a selectivity for P4VP of 2.2:1 and suggest that the position of the nitrogen in the aromatic ring of P4VP plays a key role in this selectivity. In situ plasma etching and x-ray photoelectron spectrometry measurements were made without breaking vacuum, confirming that the nitrogen plasma has selectivity for removal of P4VP over PS.

Radiation sensitivity of foodborne pathogens in meat byproducts with different packaging

NASA Astrophysics Data System (ADS)

Yong, Hae In; Kim, Hyun-Joo; Nam, Ki Chang; Kwon, Joong Ho; Jo, Cheorun

2015-10-01

The aim of this study was to determine radiation sensitivity of Escherichia coli O157:H7 and Listeria monocytogenes in edible meat byproducts. Seven beef byproducts (heart, liver, lung, lumen, omasum, large intestine, and small intestine) and four pork byproducts (heart, large intestine, liver, and small intestine) were used. Electron beam irradiation significantly reduced the numbers of pathogenic microorganisms in meat byproducts and no viable cells were detected in both aerobically- and vacuum-packaged samples irradiated at 4 kGy. Meat byproducts packed under vacuum had higher D10 value than the ones packed aerobically. No significant difference was observed between the D10 values of E. coli O157:H7 and L. monocytogenes inoculated in either aerobically or vacuum packaged samples. These results suggest that low-dose electron beam irradiation can significantly decrease microbial numbers and reduce the risk of meat byproduct contamination by the foodborne pathogens.

A Lateral Differential Resonant Pressure Microsensor Based on SOI-Glass Wafer-Level Vacuum Packaging.

PubMed

Xie, Bo; Xing, Yonghao; Wang, Yanshuang; Chen, Jian; Chen, Deyong; Wang, Junbo

2015-09-21

This paper presents the fabrication and characterization of a resonant pressure microsensor based on SOI-glass wafer-level vacuum packaging. The SOI-based pressure microsensor consists of a pressure-sensitive diaphragm at the handle layer and two lateral resonators (electrostatic excitation and capacitive detection) on the device layer as a differential setup. The resonators were vacuum packaged with a glass cap using anodic bonding and the wire interconnection was realized using a mask-free electrochemical etching approach by selectively patterning an Au film on highly topographic surfaces. The fabricated resonant pressure microsensor with dual resonators was characterized in a systematic manner, producing a quality factor higher than 10,000 (~6 months), a sensitivity of about 166 Hz/kPa and a reduced nonlinear error of 0.033% F.S. Based on the differential output, the sensitivity was increased to two times and the temperature-caused frequency drift was decreased to 25%.

A Lateral Differential Resonant Pressure Microsensor Based on SOI-Glass Wafer-Level Vacuum Packaging

PubMed Central

Xie, Bo; Xing, Yonghao; Wang, Yanshuang; Chen, Jian; Chen, Deyong; Wang, Junbo

2015-01-01

This paper presents the fabrication and characterization of a resonant pressure microsensor based on SOI-glass wafer-level vacuum packaging. The SOI-based pressure microsensor consists of a pressure-sensitive diaphragm at the handle layer and two lateral resonators (electrostatic excitation and capacitive detection) on the device layer as a differential setup. The resonators were vacuum packaged with a glass cap using anodic bonding and the wire interconnection was realized using a mask-free electrochemical etching approach by selectively patterning an Au film on highly topographic surfaces. The fabricated resonant pressure microsensor with dual resonators was characterized in a systematic manner, producing a quality factor higher than 10,000 (~6 months), a sensitivity of about 166 Hz/kPa and a reduced nonlinear error of 0.033% F.S. Based on the differential output, the sensitivity was increased to two times and the temperature-caused frequency drift was decreased to 25%. PMID:26402679

Transmission of broad W/Rh and W/Al (target/filter) x-ray beams operated at 25-49 kVp through common shielding materials.

PubMed

Li, Xinhua; Zhang, Da; Liu, Bob

2012-07-01

To provide transmission data for broad 25-39 kVp (kilovolt peak) W/Rh and 25-49 kVp W/Al (target/filter, W-tungsten, Rh-rhodium, and Al-aluminum) x-ray beams through common shielding materials, such as lead, concrete, gypsum wallboard, wood, steel, and plate glass. The unfiltered W-target x-ray spectra measured on a Selenia Dimensions system (Hologic Inc., Bedford, MA) set at 20-49 kVp were, respectively, filtered using 50-μm Rh and 700-μm Al, and were subsequently used for Monte Carlo calculations. The transmission of broad x-ray beams through shielding materials was simulated using Geant4 low energy electromagnetic physics package with photon- and electron-processes above 250 eV, including photoelectric effect, Compton scattering, and Rayleigh scattering. The calculated transmission data were fitted using Archer equation with a robust fitting algorithm. The transmission of broad x-ray beams through the above-mentioned shielding materials was calculated down to about 10(-5) for 25-39 kVp W/Rh and 25-49 kVp W/Al. The fitted results of α, β, and γ in Archer equation were provided. The α values of kVp ≥ 40 were approximately consistent with those of NCRP Report No. 147. These data provide inputs for the shielding designs of x-ray imaging facilities with W-anode x-ray beams, such as from Selenia Dimensions.

A miniature low-cost LWIR camera with a 160×120 microbolometer FPA

NASA Astrophysics Data System (ADS)

Tepegoz, Murat; Kucukkomurler, Alper; Tankut, Firat; Eminoglu, Selim; Akin, Tayfun

2014-06-01

This paper presents the development of a miniature LWIR thermal camera, MSE070D, which targets value performance infrared imaging applications, where a 160x120 CMOS-based microbolometer FPA is utilized. MSE070D features a universal USB interface that can communicate with computers and some particular mobile devices in the market. In addition, it offers high flexibility and mobility with the help of its USB powered nature, eliminating the need for any external power source, thanks to its low-power requirement option. MSE070D provides thermal imaging with its 1.65 inch3 volume with the use of a vacuum packaged CMOS-based microbolometer type thermal sensor MS1670A-VP, achieving moderate performance with a very low production cost. MSE070D allows 30 fps thermal video imaging with the 160x120 FPA size while resulting in an NETD lower than 350 mK with f/1 optics. It is possible to obtain test electronics and software, miniature camera cores, complete Application Programming Interfaces (APIs) and relevant documentation with MSE070D, as MikroSens want to help its customers to evaluate its products and to ensure quick time-to-market for systems manufacturers.

Modified-atmosphere storage under subatmospheric pressure and beef quality: II. Color, drip, cooking loss, sarcomere length, and tenderness.

PubMed

Smulders, F J M; Hiesberger, J; Hofbauer, P; Dögl, B; Dransfield, E

2006-09-01

Beef has a requirement for refrigerated storage up to 14 d to achieve adequate aging and a tender product. To achieve this aging with little spoilage and no surface drying, vacuum packaging is attractive, because it is inherently simple and offers a clear indication to the packer when the process has failed or there is risk of spoilage. However, there is increasing pressure on the meat industry to limit the use of packaging materials in view of their cost and the cost involved in their recovery and recycling. The purpose of this report was to evaluate an alternative storage system in containers using modified atmospheres at reduced pressure (approximately 25 kPa). The quality of the meat for both container- and vacuum-packed treatments was measured during chilled storage for up to 3 wk. Storage time had the most significant effect on quality characteristics, irrespective of the packaging method. Storage in containers under a 70%N2:30%CO2 gas mixture gave characteristics similar to beef stored under vacuum. Storage in containers under 100% CO2 produced less drip loss than under 70%N2:30%CO2, but generally container storage produced 3 times as much drip loss as vacuum packaging. Shear force of the LM was unaffected by the type of packaging, and at d 2 after slaughter (i.e., before the storage trial was begun), sarcomere lengths of muscles intended for container storage were similar to those destined for vacuum storage. During the packaging treatment, the comparison between the storage systems was always done within 1 animal using one carcass-half for container storage and the other half for vacuum packaging; all bulls were shackled from the left hindleg during bleeding. The majority of the muscles from the left sides had lower shear force values than those from the right sides at the earlier storage times (2 and 9 d after slaughter) but had similar values after longer storage (16 and 23 d after slaughter). This is the first report that shackling beef carcasses from the left side can result in more tender meat in the LM from that side. The increased tenderness in the LM from the shackled side probably resulted from an early decrease in pH and an increase in calpain activity after mechanical strain of the muscles on the shackled side. This effect of shackling should be taken into account when designing systematic comparisons of tenderness in beef.

Type of packaging affects the colour stability of vitamin E enriched beef.

PubMed

Nassu, Renata T; Uttaro, Bethany; Aalhus, Jennifer L; Zawadski, Sophie; Juárez, Manuel; Dugan, Michael E R

2012-12-01

Colour stability is a very important parameter for meat retail display, as appearance of the product is the deciding factor for consumers at time of purchase. This study investigated the possibility of extending appearance shelf-life through the combined use of packaging method (overwrapping - OVER, modified atmosphere - MAP, vacuum skin packaging - VSP and a combination of modified atmosphere and vacuum skin packaging - MAPVSP) and antioxidants (vitamin E enriched beef). Retail attributes (appearance, lean colour, % surface discolouration), as well as colour space analysis of images for red, green and blue parameters were measured over 18days. MAPVSP provided the most desirable retail appearance during the first 4days of retail display, while VSP-HB had the best colour stability. Overall, packaging type was more influential than α-tocopherol levels on meat colour stability, although α-tocopherol levels (>4μgg(-1) meat) had a protective effect when using high oxygen packaging methods. Crown Copyright © 2012. Published by Elsevier Ltd. All rights reserved.

Assembly/disassembly of a complex icosahedral virus to incorporate heterologous nucleic acids

NASA Astrophysics Data System (ADS)

Pascual, Elena; Mata, Carlos P.; Carrascosa, José L.; Castón, José R.

2017-12-01

Hollow protein containers are widespread in nature, and include virus capsids as well as eukaryotic and bacterial complexes. Protein cages are studied extensively for applications in nanotechnology, nanomedicine and materials science. Their inner and outer surfaces can be modified chemically or genetically, and the internal cavity can be used to template, store and/or arrange molecular cargos. Virus capsids and virus-like particles (VLP, noninfectious particles) provide versatile platforms for nanoscale bioengineering. Study of capsid protein self-assembly into monodispersed particles, and of VLP structure and biophysics is necessary not only to understand natural processes, but also to infer how these platforms can be redesigned to furnish novel functional VLP. Here we address the assembly dynamics of infectious bursal disease virus (IBDV), a complex icosahedral virus. IBDV has a ~70 nm-diameter T  =  13 capsid with VP2 trimers as the only structural subunits. During capsid assembly, VP2 is synthesized as a precursor (pVP2) whose C terminus is cleaved. The pVP2 C terminus has an amphipathic helix that controls VP2 polymorphism. In the absence of the VP3 scaffolding protein, necessary for control of assembly, 466/456-residue pVP2 intermediates bearing this helix assemble into VLP only when expressed with an N-terminal His6 tag (the HT-VP2-466 protein). HT-VP2-466 capsids are optimal for genetic insertion of proteins (cargo space ~78 000 nm3). We established an in vitro assembly/disassembly system of HT-VP2-466-based VLP for heterologous nucleic acid packaging and/or encapsulation of drugs and other molecules. HT-VP2-466 (empty) capsids were disassembled and reassembled by dialysis against low-salt/basic pH and high-salt/acid pH buffers, respectively, thus illustrating the reversibility in vitro of IBDV capsid assembly. HT-VP2-466 VLP also packed heterologous DNA by non-specific confinement during assembly. These and previous results establish the bases for biotechnological applications based on the IBDV capsid and its ability to incorporate exogenous proteins and nucleic acids.

Elucidation of the Block to Herpes Simplex Virus Egress in the Absence of Tegument Protein UL16 Reveals a Novel Interaction with VP22

PubMed Central

Starkey, Jason L.; Han, Jun; Chadha, Pooja; Marsh, Jacob A.

2014-01-01

UL16 is a tegument protein of herpes simplex virus (HSV) that is conserved among all members of the Herpesviridae, but its function is poorly understood. Previous studies revealed that UL16 is associated with capsids in the cytoplasm and interacts with the membrane protein UL11, which suggested a “bridging” function during cytoplasmic envelopment, but this conjecture has not been tested. To gain further insight, cells infected with UL16-null mutants were examined by electron microscopy. No defects in the transport of capsids to cytoplasmic membranes were observed, but the wrapping of capsids with membranes was delayed. Moreover, clusters of cytoplasmic capsids were often observed, but only near membranes, where they were wrapped to produce multiple capsids within a single envelope. Normal virion production was restored when UL16 was expressed either by complementing cells or from a novel position in the HSV genome. When the composition of the UL16-null viruses was analyzed, a reduction in the packaging of glycoprotein E (gE) was observed, which was not surprising, since it has been reported that UL16 interacts with this glycoprotein. However, levels of the tegument protein VP22 were also dramatically reduced in virions, even though this gE-binding protein has been shown not to depend on its membrane partner for packaging. Cotransfection experiments revealed that UL16 and VP22 can interact in the absence of other viral proteins. These results extend the UL16 interaction network beyond its previously identified binding partners to include VP22 and provide evidence that UL16 plays an important function at the membrane during virion production. PMID:24131716

Modification of in-pack conditions to extend the storage life of vacuum packaged lamb.

PubMed

Gill, C O; Penney, N

1985-01-01

High pH (>5·9) lamb loins from a research abattoir were subjected to differing packaging treatments to determine whether package modification could reliably extend the storage life of chilled lamb cuts beyond that attained by cuts vacuum-packaged in film of low gas permeability, as in current commercial practice. Treatments applied were carbon dioxide flushing or addition of a citrate buffer (pH 4·8), a 5% lactic acid solution or a Lactobacillus inoculum (plastic packs only) and packaging in a plastic film of moderately low oxygen permeability (140 cc/m(2)/24 h at 25°C and 90% relative humidity) or in a foil laminate of immeasurably low oxygen permeability. After 12 weeks' storage at -0.5°C, the cuts packaged in the plastic film were spoiled by off-odours produced by enterobacteria, except for inoculated cuts, which, instead, had developed unacceptable dairy flavours. In contrast, cuts packaged in foil laminate developed floras of lactobacilli that had not caused spoilage after 12 weeks, and meat colour was much improved by the exclusion of oxygen. Loin cuts from a commercial packaging operation were packaged in a shrinkable plastic film of low oxygen permeability (30 to 40 cc/m(2)/24 h at 25°C and 90% relative humidity), in foil laminate, or in foil laminate after the addition of 5% lactic acid solution. For the first 6 weeks, cuts were stored in a commercial chiller nominally operating at 0°C; subsequently, they were held in a laboratory chiller at -0.5°C. Some cuts packaged in the shrinkable plastic were spoiled after 9 weeks' storage and all were spoiled at 12 weeks, because of off-flavours produced by enterobacteria. All cuts packaged in the foil laminate were very acceptable at 9 weeks but most were spoiled by off-flavours at 12 weeks. Most cuts treated with lactic acid and packaged in foil laminate were unspoiled after 12 weeks. The packaging requirements indicated to be necessary for reliable extension of the storage life of vacuum packaged lamb are discussed. Copyright © 1985. Published by Elsevier Ltd.

320 x 240 uncooled IRFPA with pixel wise thin film vacuum packaging

NASA Astrophysics Data System (ADS)

Yon, J.-J.; Dumont, G.; Rabaud, W.; Becker, S.; Carle, L.; Goudon, V.; Vialle, C.; Hamelin, A.; Arnaud, A.

2012-10-01

Silicon based vacuum packaging is a key enabling technology for achieving affordable uncooled Infrared Focal Plane Arrays (IRFPA) as required by the promising mass market for very low cost IR applications, such as automotive driving assistance, energy loss monitoring in buildings, motion sensors… Among the various approaches studied worldwide, the CEA, LETI is developing a unique technology where each bolometer pixel is sealed under vacuum at the wafer level, using an IR transparent thin film deposition. This technology referred to as PLP (Pixel Level Packaging), leads to an array of hermetic micro-caps each containing a single microbolometer. Since the successful demonstration that the PLP technology, when applied on a single microbolometer pixel, can provide the required vacuum < 10-3 mbar, the authors have pushed forward the development of the technology on fully operational QVGA readout circuits CMOS base wafers (320 x 240 pixels). In this outlook, the article reports on the electro optical performance obtained from this preliminary PLP based QVGA demonstrator. Apart from the response, noise and NETD distributions, the paper also puts emphasis on additional key features such as thermal time constant, image quality, and ageing properties.

Use of antimicrobial biodegradable packaging to control Listeria monocytogenes during storage of cooked ham.

PubMed

Marcos, Begonya; Aymerich, Teresa; Monfort, Josep M; Garriga, Margarita

2007-11-30

The antimicrobial effect against L. monocytogenes of biodegradable films (alginate, zein and polyvinyl alcohol) containing enterocins was investigated. Survival of the pathogen was studied by means of challenge tests performed at 6 degrees C during 8 and 29 days, for air-packed and vacuum-packed sliced cooked ham, respectively. Air packaging was tested with two concentrations of enterocins (200 and 2000 AU/cm2). Control air-packed cooked ham showed an increase of L. monocytogenes from 10(4) to 10(7) CFU/g after 8 days. By contrast, packaging with antimicrobial films effectively slowed down the pathogen's growth, leading to final counts lower than in control lots. Air-packaging with alginate films containing 2000 AU/cm2 of enterocins effectively controlled L. monocytogenes for 8 days. An increase of only 1 log unit was observed in zein and polyvinyl alcohol lots at the same enterocin concentration. Vacuum packaging with films containing enterocins (2000 AU/cm2) also delayed the growth of the pathogen. No increase from inoculated levels was observed during 15 days in antimicrobial alginate films. After 29 days of storage, the lowest counts were obtained in samples packed with zein and alginate films containing enterocins, as well as with zein control films. The most effective treatment for controlling L. monocytogenes during 6 degrees C storage was vacuum-packaging of sliced cooked ham with alginate films containing 2000 AU/cm2 of enterocins. From the results obtained it can concluded that antimicrobial packaging can improve the safety of sliced cooked ham by delaying and reducing the growth of L. monocytogenes.

Transmission of broad W/Rh and W/Al (target/filter) x-ray beams operated at 25-49 kVp through common shielding materials

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

Li Xinhua; Zhang Da; Liu, Bob

2012-07-15

Purpose: To provide transmission data for broad 25-39 kVp (kilovolt peak) W/Rh and 25-49 kVp W/Al (target/filter, W-tungsten, Rh-rhodium, and Al-aluminum) x-ray beams through common shielding materials, such as lead, concrete, gypsum wallboard, wood, steel, and plate glass. Methods: The unfiltered W-target x-ray spectra measured on a Selenia Dimensions system (Hologic Inc., Bedford, MA) set at 20-49 kVp were, respectively, filtered using 50-{mu}m Rh and 700-{mu}m Al, and were subsequently used for Monte Carlo calculations. The transmission of broad x-ray beams through shielding materials was simulated using Geant4 low energy electromagnetic physics package with photon- and electron-processes above 250 eV,more » including photoelectric effect, Compton scattering, and Rayleigh scattering. The calculated transmission data were fitted using Archer equation with a robust fitting algorithm. Results: The transmission of broad x-ray beams through the above-mentioned shielding materials was calculated down to about 10{sup -5} for 25-39 kVp W/Rh and 25-49 kVp W/Al. The fitted results of {alpha}, {beta}, and {gamma} in Archer equation were provided. The {alpha} values of kVp Greater-Than-Or-Slanted-Equal-To 40 were approximately consistent with those of NCRP Report No. 147. Conclusions: These data provide inputs for the shielding designs of x-ray imaging facilities with W-anode x-ray beams, such as from Selenia Dimensions.« less

Comparison of the Transcription and Replication Strategies of Marburg Virus and Ebola Virus by Using Artificial Replication Systems

PubMed Central

Mühlberger, Elke; Weik, Michael; Volchkov, Viktor E.; Klenk, Hans-Dieter; Becker, Stephan

1999-01-01

The members of the family Filoviridae, Marburg virus (MBGV) and Ebola virus (EBOV), are very similar in terms of morphology, genome organization, and protein composition. To compare the replication and transcription strategies of both viruses, an artificial replication system based on the vaccinia virus T7 expression system was established for EBOV. Specific transcription and replication of an artificial monocistronic minireplicon was demonstrated by reporter gene expression and detection of the transcribed and replicated RNA species. As it was shown previously for MBGV, three of the four EBOV nucleocapsid proteins, NP, VP35, and L, were essential and sufficient for replication. In contrast to MBGV, EBOV-specific transcription was dependent on the presence of the fourth nucleocapsid protein, VP30. When EBOV VP30 was replaced by MBGV VP30, EBOV-specific transcription was observed but with lower efficiency. Exchange of NP, VP35, and L between the two replication systems did not lead to detectable reporter gene expression. It was further observed that neither MBGV nor EBOV were able to replicate the heterologous minigenomes. A chimeric minigenome, however, containing the EBOV leader and the MBGV trailer was encapsidated, replicated, transcribed, and packaged by both viruses. PMID:9971816

Ultraminiature Broadband Light Source and Method of Manufacturing Same

NASA Technical Reports Server (NTRS)

Tuma, Margaret L. (Inventor); Collura, Joseph S. (Inventor); Helvajian, Henry (Inventor); Pocha, Michael D. (Inventor); Meyer, Glenn A. (Inventor); McConaghy, Charles F. (Inventor); Olsen, Barry L. (Inventor); Hansen, William W. (Inventor)

2010-01-01

An ultraminiature light source using a double-spiral shaped tungsten filament includes end contact portions which are separated to allow for radial and length-wise unwinding of the spiral. The double-spiral filament is spaced relatively far apart at the end portions thereof so that contact between portions of the filament upon expansion is avoided. The light ource is made by fabricating a double-spiral ultraminiature tungsten filament from tungsten foil and housing the filament in a ceramic package having a reflective bottom and a well wherein the filament is suspended. A vacuum furnace brazing process attaches the filament to contacts of the ceramic package. Finally, a cover with a transparent window is attached onto the top of the ceramic package by solder reflow in a second vacuum furnace process to form a complete hermetically sealed package.

Ultraminiature broadband light source with spiral shaped filament

NASA Technical Reports Server (NTRS)

McConaghy, Charles F. (Inventor); Olsen, Barry L. (Inventor); Tuma, Margaret L. (Inventor); Collura, Joseph S. (Inventor); Pocha, Michael D. (Inventor); Helvajian, Henry (Inventor); Meyer, Glenn A. (Inventor); Hansen, William W (Inventor)

2012-01-01

An ultraminiature light source using a double-spiral shaped tungsten filament includes end contact portions which are separated to allow for radial and length-wise unwinding of the spiral. The double-spiral filament is spaced relatively far apart at the end portions thereof so that contact between portions of the filament upon expansion is avoided. The light source is made by fabricating a double-spiral ultraminiature tungsten filament from tungsten foil and housing the filament in a ceramic package having a reflective bottom and a well wherein the filament is suspended. A vacuum furnace brazing process attaches the filament to contacts of the ceramic package. Finally, a cover with a transparent window is attached onto the top of the ceramic package by solder reflow in a second vacuum furnace process to form a complete hermetically sealed package.

Effect of including whole linseed and vitamin E in the diet of young bulls slaughtered at two fat covers on the sensory quality of beef packaged in two different packaging systems.

PubMed

Albertí, Pere; Campo, María M; Beriain, María J; Ripoll, Guillermo; Sañudo, Carlos

2017-02-01

Forty-six Pirenaica young bulls, slaughtered at two levels of fatness (3 and 4 mm), were used to evaluate the effect of the inclusion of 50 g kg -1 linseed alone or with 200 IU vitamin E kg -1 in the concentrate and of the meat packaging system (vacuum or modified atmosphere packaging (MAP)) on the beef sensory quality. The inclusion of linseed or supplementation with vitamin E in the concentrate induced no significant differences in the main meat sensory scores and overall appraisal except under MAP, where small differences due to concentrate ingredients were found in juiciness and metallic flavor intensity. Extending the display time up to 4 or 8 days in high-oxygen MAP had detrimental effects on sensory attributes. Meat from animals with 4 mm fat cover depth were rated more tender and juicy, less fibrous and with a higher intensity of beef flavor and rancid odor than meat from 3 mm fat cover bulls when both samples were vacuum packaged. The inclusion of 50 g kg -1 linseed in the concentrate fed to bulls had no detrimental effect on the beef sensory quality. The vacuum-packaged meat of bulls slaughtered at 4 mm fat cover was rated higher on sensory analysis than that at 3 mm fat cover. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

Carbon monoxide concentration and exposure time effects on the depth of CO penetration and surface color of raw and cooked beef longissimus lumborum steaks.

PubMed

Sakowska, A; Guzek, D; Głąbska, D; Wierzbicka, A

2016-11-01

This study investigated the influence of carbon monoxide (CO) exposure time (0, 7, 14, and 21days) and concentration in gas mixture on depth of penetration and the surface color of raw and cooked striploin steaks. Seven packaging treatments were evaluated: vacuum, vacuum after 48h of exposure to 0.1%, 0.3% or 0.5% CO (mixed with 30% CO2 and 69.5-69.9% N2), and modified atmosphere packaging (MAP) containing the same gas mixtures. CO penetration depth increased as exposure times and CO concentration in gas mixtures increased (p<0.05). However, the carboxymyoglobin that formed did not always turn brown during thermal treatment. In cooked samples treated with 0.3% and 0.5% CO-MAP, a red carboxymyoglobin border was visible at the cross section, whereas other CO packaging treatments had its partial or total browning. To create a red color in raw and avoid a red boarder in cooked beef, up to 0.5% CO in vacuum packages and only 0.1% for MAP can be recommended. Copyright © 2016 Elsevier Ltd. All rights reserved.

Chemical composition of the continental crust: Insights from a quantitative interpretation of the Vp/Vs ratio

NASA Astrophysics Data System (ADS)

Guerri, M.; Youssof, M.; Fullea, J.

2017-12-01

The processes driving continental crust formation are not yet fully understood. One of the fundamental keys necessary to investigate the enigma is represented by crustal composition. The Vp/Vs ratio from seismic receiver functions or tomography studies is a powerful tool to constrain the crustal composition. However, to date only qualitative relationships between Vp/Vs and composition have been proposed. We present a quantitative interpretation of the Vp/Vs in terms of major oxide components, based on thermo-elastic constrained modelling of rock phase equilibria and physical properties. The geophysical-petrological approach is implemented in the new release of the software package LitMod, which now allows for integrated and self-consistent modeling of the entire lithosphere (crust + lithospheric mantle) and upper mantle. Forward modelling of the Vp/Vs, based on petrology and thermodynamics, reveals that, as expected, mafic compositions have higher Vp/Vs than felsic ones. However, in high temperature settings (surface heat flow > 75 mW/m2), the quartz alpha / quartz beta transition strongly increases Vp, leaving Vs almost unaltered, leading to SiO2-rich compositions displaying Vp/Vs values higher than those associated with mafic compositions. Additionally, we highlight the importance of H2O, the presence of which stabilizes amphibole (in place of pyroxene), characterized by a relatively low Vp/Vs. If H2O is present, mafic compositions show Vp/Vs ratios that are comparable to those produced by anhydrous SiO2-rich compositions. The destabilization of amphibole (in favour of pyroxene) generates a sharp seismic discontinuity, potentially detectable by, for example, seismic refraction and receiver function investigations. We invert the Vp/Vs ratio for composition and hydrous state of the crust in the Southern African cratons. Our results show that the Kaapvaal craton, Archean in age, has an intermediate (SiO2 60 wt%) composition. The finding has implications on our understanding of the mechanisms responsible for crustal formation as an intermediate bulk composition is potentially generated by subduction-related processes. We conclude that, already before the Meso-Archean (3.2 Ga), subduction related crust-forming processes were active in the area and potentially also in other cratons.

Effects of self-carbon dioxide-generation material for active packaging on pH, water-holding capacity, meat color, lipid oxidation and microbial growth in beef during cold storage.

PubMed

Lee, Seung-Jae; Lee, Seung Yun; Kim, Gap-Don; Kim, Geun-Bae; Jin, Sang Keun; Hur, Sun Jin

2017-08-01

Active packaging refers to the mixing of additive agents into packaging materials with the purpose of maintaining or extending food product quality and shelf life. The aim of this study was to develop an easy and cheap active packaging for beef. Beef loin samples were divided into three packaging groups (C, ziplock bag packaging; T1, vacuum packaging; T2, active packaging) and stored at 4 °C for 21 days. The water-holding capacity was significantly (P < 0.05) higher in C and T2 than in T1 for up to 7 days of storage. The TBARS value was significantly (P < 0.05) lower in T1 and T2 after 7 days of storage. The counts of some microorganism were significantly (P < 0.05) lower in T1 and T2 after 7 days of storage; the total bacterial count and Escherichia coli count were lowest in T2 at the end of storage. These results indicate that active packaging using self-CO 2 -generation materials can extend the shelf life similarly to that observed with vacuum packaging, and that the active packaging method can improve the quality characteristics of beef during cold storage. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

9 CFR 590.548 - Drying, blending, packaging, and heat treatment rooms and facilities.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 9 Animals and Animal Products 2 2011-01-01 2011-01-01 false Drying, blending, packaging, and heat..., blending, packaging, and heat treatment rooms and facilities. (a) General. Processing rooms shall be... vacuum cleaned daily. (c) The heat treatment room shall be of an approved construction and be maintained...

9 CFR 590.548 - Drying, blending, packaging, and heat treatment rooms and facilities.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 9 Animals and Animal Products 2 2013-01-01 2013-01-01 false Drying, blending, packaging, and heat..., blending, packaging, and heat treatment rooms and facilities. (a) General. Processing rooms shall be... vacuum cleaned daily. (c) The heat treatment room shall be of an approved construction and be maintained...

9 CFR 590.548 - Drying, blending, packaging, and heat treatment rooms and facilities.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 9 Animals and Animal Products 2 2012-01-01 2012-01-01 false Drying, blending, packaging, and heat..., blending, packaging, and heat treatment rooms and facilities. (a) General. Processing rooms shall be... vacuum cleaned daily. (c) The heat treatment room shall be of an approved construction and be maintained...

9 CFR 590.548 - Drying, blending, packaging, and heat treatment rooms and facilities.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 9 Animals and Animal Products 2 2014-01-01 2014-01-01 false Drying, blending, packaging, and heat..., blending, packaging, and heat treatment rooms and facilities. (a) General. Processing rooms shall be... vacuum cleaned daily. (c) The heat treatment room shall be of an approved construction and be maintained...

Multiple internal seal right micro-electro-mechanical system vacuum package

NASA Technical Reports Server (NTRS)

Shcheglov, Kirill V. (Inventor); Wiberg, Dean V. (Inventor); Hayworth, Ken J. (Inventor); Yee, Karl Y. (Inventor); Bae, Youngsam (Inventor); Challoner, A. Dorian (Inventor); Peay, Chris S. (Inventor)

2007-01-01

A Multiple Internal Seal Ring (MISR) Micro-Electro-Mechanical System (MEMS) vacuum package that hermetically seals MEMS devices using MISR. The method bonds a capping plate having metal seal rings to a base plate having metal seal rings by wafer bonding the capping plate wafer to the base plate wafer. Bulk electrodes may be used to provide conductive paths between the seal rings on the base plate and the capping plate. All seals are made using only metal-to-metal seal rings deposited on the polished surfaces of the base plate and capping plate wafers. However, multiple electrical feed-through metal traces are provided by fabricating via holes through the capping plate for electrical connection from the outside of the package through the via-holes to the inside of the package. Each metal seal ring serves the dual purposes of hermetic sealing and providing the electrical feed-through metal trace.

Multiple internal seal ring micro-electro-mechanical system vacuum packaging method

NASA Technical Reports Server (NTRS)

Hayworth, Ken J. (Inventor); Yee, Karl Y. (Inventor); Shcheglov, Kirill V. (Inventor); Bae, Youngsam (Inventor); Wiberg, Dean V. (Inventor); Challoner, A. Dorian (Inventor); Peay, Chris S. (Inventor)

2008-01-01

A Multiple Internal Seal Ring (MISR) Micro-Electro-Mechanical System (MEMS) vacuum packaging method that hermetically seals MEMS devices using MISR. The method bonds a capping plate having metal seal rings to a base plate having metal seal rings by wafer bonding the capping plate wafer to the base plate wafer. Bulk electrodes may be used to provide conductive paths between the seal rings on the base plate and the capping plate. All seals are made using only metal-to-metal seal rings deposited on the polished surfaces of the base plate and capping plate wafers. However, multiple electrical feed-through metal traces are provided by fabricating via holes through the capping plate for electrical connection from the outside of the package through the via-holes to the inside of the package. Each metal seal ring serves the dual purposes of hermetic sealing and providing the electrical feed-through metal trace.

Characterization of a potentially novel 'blown pack' spoilage bacterium isolated from bovine hide.

PubMed

Moschonas, G; Bolton, D J

2013-03-01

To characterize a psychrotrophic bacterium, designated TC1, previously isolated from a cattle hide in Ireland, and to investigate the ability of this strain to cause 'blown pack' spoilage (BPS) of vacuum-packaged beef primals. TC1 was characterized using a combination of phenotypic, chemotaxonomic and genotypic analyses and was assessed for its ability to spoil vacuum-packaged beef at refrigerated temperatures. TC1 was Gram-positive and formed elliptical subterminal endospores. The strain was able to grow between 0 and 33 °C, with optimal growth between 23 and 24 °C. TC1 could be differentiated from its phylogenetically closest neighbour (Clostridium lituseburense DSM 797(T)) by 16S rRNA gene sequencing, pulsed-field gel electrophoresis and cellular fatty acid composition. TC1 spoiled (BPS) beef within 42 days when inoculated in cold-stored (1 °C) vacuum-packed beef. The phenotypic, chemotaxonomic and genotypic characterization indicated that TC1 may represent a potentially novel, cold-tolerant, gas-producing bacterium of considerable economic significance to the beef industry. This study reports and characterizes an emerging BPS bacterium, which should be considered in future activities designed to minimize the psychrophilic and psychrotrophic spoilage of vacuum-packaged beef. © 2012 The Society for Applied Microbiology.

A Novel Method to Decontaminate Surgical Instruments for Operational and Austere Environments.

PubMed

Knox, Randy W; Demons, Samandra T; Cunningham, Cord W

2015-12-01

The purpose of this investigation was to test a field-expedient, cost-effective method to decontaminate, sterilize, and package surgical instruments in an operational (combat) or austere environment using chlorhexidine sponges, ultraviolet C (UVC) light, and commercially available vacuum sealing. This was a bench study of 4 experimental groups and 1 control group of 120 surgical instruments. Experimental groups were inoculated with a 10(6) concentration of common wound bacteria. The control group was vacuum sealed without inoculum. Groups 1, 2, and 3 were first scrubbed with a chlorhexidine sponge, rinsed, and dried. Group 1 was then packaged; group 2 was irradiated with UVC light, then packaged; group 3 was packaged, then irradiated with UVC light through the bag; and group 4 was packaged without chlorhexidine scrubbing or UVC irradiation. The UVC was not tested by itself, as it does not grossly clean. The instruments were stored overnight and tested for remaining colony forming units (CFU). Data analysis was conducted using analysis of variance and group comparisons using the Tukey method. Group 4 CFU was statistically greater (P < .001) than the control group and groups 1 through 3. There was no statistically significant difference between the control group and groups 1 through 3. Vacuum sealing of chlorhexidine-scrubbed contaminated instruments with and without handheld UVC irradiation appears to be an acceptable method of field decontamination. Chlorhexidine scrubbing alone achieved a 99.9% reduction in CFU, whereas adding UVC before packaging achieved sterilization or 100% reduction in CFU, and UVC through the bag achieved disinfection. Published by Elsevier Inc.

Vacuum decay container/closure integrity testing technology. Part 1. ASTM F2338-09 precision and bias studies.

PubMed

Wolf, Heinz; Stauffer, Tony; Chen, Shu-Chen Y; Lee, Yoojin; Forster, Ronald; Ludzinski, Miron; Kamat, Madhav; Godorov, Phillip; Guazzo, Dana Morton

2009-01-01

ASTM F2338-09 Standard Test Method for Nondestructive Detection of Leaks in Packages by Vacuum Decay Method is applicable for leak-testing rigid and semi-rigid non-lidded trays; trays or cups sealed with porous barrier lidding materials; rigid, nonporous packages; and flexible, nonporous packages. Part 1 of this series describes the precision and bias studies performed in 2008 to expand this method's scope to include rigid, nonporous packages completely or partially filled with liquid. Round robin tests using three VeriPac 325/LV vacuum decay leak testers (Packaging Technologies & Inspection, LLC, Tuckahoe, NY) were performed at three test sites. Test packages were 1-mL glass syringes. Positive controls had laser-drilled holes in the barrel ranging from about 5 to 15 microm in nominal diameter. Two different leak tests methods were performed at each site: a "gas leak test" performed at 250 mbar (absolute) and a "liquid leak test" performed at about 1 mbar (absolute). The gas leak test was used to test empty, air-filled syringes. All defects with holes > or = 5.0 microm and all no-defect controls were correctly identified. The only false negative result was attributed to a single syringe with a < 5.0-microm hole. Tests performed using a calibrated air leak supported a 0.10-cm3 x min(-1) (ccm) sensitivity limit (99/99 lower tolerance limit). The liquid leak test was used to test both empty, air-filled syringes and water-filled syringes. Test results were 100% accurate for all empty and water-filled syringes, both without holes and with holes (5, 10, and 15 microm). Tests performed using calibrated air flow leaks of 0, 0.05, and 0.10 ccm were also 100% accurate; data supported a 0.10-ccm sensitivity limit (99/99 lower tolerance limit). Quantitative differential pressure results strongly correlated to hole size using either liquid or gas vacuum decay leak tests. The higher vacuum liquid leak test gave noticeably higher pressure readings when water was present in the defect. Both the ASTM F2338-09 test method and the precision and bias study report are available by contacting ASTM International in West Conshohocken, PA, USA (www.astm.org).

Effects of vacuum and modified atmosphere on textural parameters and structural proteins of cultured meagre (Argyrosomus regius) fillets.

PubMed

Sáez, María I; Martínez, Tomás F; Cárdenas, Salvador; Suárez, María D

2015-09-01

The influence of two preservation strategies (vacuum package and modified atmosphere package) on the post-mortem changes of textural parameters, pH, water holding capacity, sarcoplasmic and myofibrillar proteins, and collagen content of meagre (Argyrosomus regius) fillets was studied. Fillets were stored in a cold room in aerobic (control, C), vacuum (V) and modified atmosphere (MA) package. Samples were withdrawn at six sampling points throughout 15-day storage, and post-mortem changes were assessed. The textural parameters were significantly enhanced in V and MA compared to C. Both V and MA treatments reduced the intensity of a group of myofibrillar protein fractions (140-195 kDa) and increased insoluble collagen compared to C. Consequently, the post-mortem flesh softening in C was attributed to increased proteolysis in both intracellular and extracellular structural proteins. The preservation of the textural and biochemical characteristics of meagre fillets subjected to V and MA treatments makes these two treatments highly recommendable for the commercialization of meagre fillets. © The Author(s) 2014.

Effect of low-temperature preservation on the quality of vacuum-packaged dry-cured ham: Refrigerated boneless ham and frozen ham cuts.

PubMed

Cilla, Irene; Martínez, Luis; Beltrán, José Antonio; Roncalés, Pedro

2006-05-01

The effect of storage on dry-cured ham quality was studied. Sixteen vacuum-packaged boneless dry-cured hams and sixteen vacuum-packaged dry-cured ham cuts were stored in darkness under refrigeration (4±2°C; 8 months) or freezing (-18±1°C; 24 months), respectively. Instrumental colour and texture, physico-chemical and biochemical parameters, sensory profile and consumer acceptability and purchase satisfaction were measured throughout storage. The overall quality of refrigerated boneless dry-cured hams and frozen dry-cured ham cuts showed only limited changes throughout long-term storage. Significant changes involved loss of odour and flavour, increased adhesiveness and modification of hardness, the Semimembranosus muscle became tender while Biceps femoris became harder, leading to a higher textural homogeneity. In agreement with those changes, the overall acceptability assessed by a trained panel decreased throughout storage, though this was significant regarding only frozen hams. However, consumer evaluation of acceptability, as well as satisfaction with hypothetical purchasing, did not vary significantly throughout storage.

Effect of packaging and storage time on survival of Listeria monocytogenes on kippered beef steak and turkey tenders.

PubMed

Uppal, Kamaldeep K; Getty, Kelly J K; Boyle, Elizabeth A E; Harper, Nigel M; Lobaton-Sulabo, April Shayne S; Barry, Bruce

2012-01-01

The objective of our study was to determine effect of packaging method and storage time on reducing Listeria monocytogenes in shelf-stable meat snacks. Commercially available kippered beef steak strips and turkey tenders were dipped into a 5-strain L. monocytogenes cocktail, and dried at 23 °C until a water activity of 0.80 was achieved. Inoculated samples were packaged with 4 treatments: (1) vacuum, (2) nitrogen flushed with oxygen scavenger, (3) heat sealed with oxygen scavenger, and (4) heat sealed without oxygen scavenger. Samples were stored at 23 °C and evaluated for L. monocytogenes levels at 0, 24, 48, and 72 h. Initial levels (time 0) of L. monocytogenes were approximately 5.7 log CFU/cm² for steak and tenders. After 24 h of storage time, a 1 log CFU/cm² reduction of L. monocytogenes was observed for turkey tenders for all packaging treatments. After 48 h, turkey tenders showed >1 log CFU/cm² reduction of L. monocytogenes for all packaging treatments except for vacuum, where only 0.9 log CFU/cm² reduction was observed. After 72 h, reductions for all packaging treatments for turkey tenders ranged from 1.5 to 2.4 log CFU/cm². For kippered beef steak, there was no interaction between the packaging treatments and all storage times (P > 0.05) whereas, time was different (P <0.05). For kippered beef steak, there was 1 log reduction of L. monocytogenes at 24 and 48 h of storage times at 23 °C for all packaging treatments and a 2.1 log CFU/ cm² L. monocytogenes reduction at 72 h of storage time. Processors of kippered beef steak and turkey tenders could use a combination of vacuum or nitrogen-flushing or heat sealed with an oxygen scavenger packaging methods and a holding time of 24 h prior to shipping to reduce potential L. monocytogenes numbers by ≥1 log. However, processors should be encouraged to hold packaged product a minimum of 72 h to enhance the margin of safety for L. monocytogenes control. © 2011 Institute of Food Technologists®

Postharvest changes in the phenolic profile of watercress induced by post-packaging irradiation and modified atmosphere packaging.

PubMed

Pinela, José; Barros, Lillian; Barreira, João C M; Carvalho, Ana Maria; Oliveira, M Beatriz P P; Santos-Buelga, Celestino; Ferreira, Isabel C F R

2018-07-15

The effects of γ-ray irradiation and modified atmosphere packaging (MAP) on watercress (Nasturtium officinale R. Br.) phenolic compounds were evaluated after 7-day storage at 4 °C. Irradiation doses of 1, 2 and 5 kGy were tested, as well as vacuum-packaging and MAP enriched with 100% N 2 and Ar. A non-irradiated, air-packaged control was included in all experiments. p-Coumaric acid was the most abundant compound in fresh watercress, followed by quercetin-3-O-sophoroside and isorhamnetin-O-hydroxyferuloylhexoside-O-hexoside. Four kaempferol glycoside derivatives were identified for the first time in this species. In general, flavonoids predominated over phenolic acids. Samples stored under vacuum and irradiated at 2 kGy revealed lower phenolic levels. Ar-enriched MAP and control conditions preserved the initial phenolic content. The 5 kGy dose also maintained concentrations of flavonoids and total phenolic compounds, but increased the phenolic acids content. Additionally, flavonoids were found strongly correlated to DPPH scavenging activity and β-carotene bleaching inhibition capacity. Copyright © 2018 Elsevier Ltd. All rights reserved.

7 CFR 58.241 - Packaging, repackaging and storage.

Code of Federal Regulations, 2012 CFR

2012-01-01

... contamination. The room should be vacuumed as often as necessary and kept clean and orderly. ... clean, sound commercially accepted container or packaging material which will satisfactorily protect the... them practically free of residual product before being transferred from the filling room to the...

7 CFR 58.241 - Packaging, repackaging and storage.

Code of Federal Regulations, 2014 CFR

2014-01-01

... contamination. The room should be vacuumed as often as necessary and kept clean and orderly. ... clean, sound commercially accepted container or packaging material which will satisfactorily protect the... them practically free of residual product before being transferred from the filling room to the...

Modified atmosphere packaging and vacuum packaging for long period chilled storage of dry-cured Iberian ham.

PubMed

Parra, V; Viguera, J; Sánchez, J; Peinado, J; Espárrago, F; Gutierrez, J I; Andrés, A I

2010-04-01

Dry-cured Iberian ham slices were stored under vacuum and under four different modified atmospheres (60/40=60%N(2)+40%CO(2); 70/30=70%N(2)+30%CO(2); 80/20=80%N(2)+20%CO(2); argon=70%argon+30%CO(2)) at 4+/-1 degrees C during 120 days. Gas composition, moisture content, pH, colour, pigment content, and lipid stability were measured, as well as sensory and microbial analysis were carried out throughout storage. A loss of intensity of red colour (a(*)-values) was observed during storage in ham slices (P<0.05). Consistently, MbFe(II)NO content also decreased throughout storage (P>0.05). Slices of ham packed in 40%CO(2) (60/40) and 30%CO(2) (70/30) showed lower a(*)-values than the rest of the batches after 60 days (P<0.05), though differences were not evident after 120 days (P>0.05). TBARs values showed an upward trend during the storage of packaged slices (P<0.05). Vacuum-packed slices showed the lowest TBARs values and those packed with 40%CO(2), the highest. Sensory attributes did not vary significantly (P>0.05) throughout storage under refrigeration and packed either in vacuum or in modified atmospheres. No safety problems were detected in relation to the microbial quality in any case. 2009 Elsevier Ltd. All rights reserved.

Reducing Salt in Raw Pork Sausages Increases Spoilage and Correlates with Reduced Bacterial Diversity

PubMed Central

Fougy, Lysiane; Desmonts, Marie-Hélène; Coeuret, Gwendoline; Fassel, Christine; Hamon, Erwann; Hézard, Bernard; Champomier-Vergès, Marie-Christine

2016-01-01

ABSTRACT Raw sausages are perishable foodstuffs; reducing their salt content raises questions about a possible increased spoilage of these products. In this study, we evaluated the influence of salt reduction (from 2.0% to 1.5% [wt/wt]), in combination with two types of packaging (modified atmosphere [50% mix of CO2-N2] and vacuum packaging), on the onset of spoilage and on the diversity of spoilage-associated bacteria. After 21 days of storage at 8°C, spoilage was easily observed, characterized by noticeable graying of the products and the production of gas and off-odors defined as rancid, sulfurous, or sour. At least one of these types of spoilage occurred in each sample, and the global spoilage intensity was more pronounced in samples stored under modified atmosphere than under vacuum packaging and in samples with the lower salt content. Metagenetic 16S rRNA pyrosequencing revealed that vacuum-packaged samples contained a higher total bacterial richness (n = 69 operational taxonomic units [OTUs]) than samples under the other packaging condition (n = 46 OTUs). The core community was composed of 6 OTUs (Lactobacillus sakei, Lactococcus piscium, Carnobacterium divergens, Carnobacterium maltaromaticum, Serratia proteamaculans, and Brochothrix thermosphacta), whereas 13 OTUs taxonomically assigned to the Enterobacteriaceae, Enterococcaceae, and Leuconostocaceae families comprised a less-abundant subpopulation. This subdominant community was significantly more abundant when 2.0% salt and vacuum packaging were used, and this correlated with a lower degree of spoilage. Our results demonstrate that salt reduction, particularly when it is combined with CO2-enriched packaging, promotes faster spoilage of raw sausages by lowering the overall bacterial diversity (both richness and evenness). IMPORTANCE Our study takes place in the context of raw meat product manufacturing and is linked to a requirement for salt reduction. Health guidelines are calling for a reduction in dietary salt intake. However, salt has been used for a very long time as a hurdle technology, and salt reduction in meat products raises the question of spoilage and waste of food. The study was conceived to assess the role of sodium chloride reduction in meat products, both at the level of spoilage development and at the level of bacterial diversity, using 16S rRNA amplicon sequencing and raw pork sausage as a meat model. PMID:27107120

Reducing Salt in Raw Pork Sausages Increases Spoilage and Correlates with Reduced Bacterial Diversity.

PubMed

Fougy, Lysiane; Desmonts, Marie-Hélène; Coeuret, Gwendoline; Fassel, Christine; Hamon, Erwann; Hézard, Bernard; Champomier-Vergès, Marie-Christine; Chaillou, Stéphane

2016-07-01

Raw sausages are perishable foodstuffs; reducing their salt content raises questions about a possible increased spoilage of these products. In this study, we evaluated the influence of salt reduction (from 2.0% to 1.5% [wt/wt]), in combination with two types of packaging (modified atmosphere [50% mix of CO2-N2] and vacuum packaging), on the onset of spoilage and on the diversity of spoilage-associated bacteria. After 21 days of storage at 8°C, spoilage was easily observed, characterized by noticeable graying of the products and the production of gas and off-odors defined as rancid, sulfurous, or sour. At least one of these types of spoilage occurred in each sample, and the global spoilage intensity was more pronounced in samples stored under modified atmosphere than under vacuum packaging and in samples with the lower salt content. Metagenetic 16S rRNA pyrosequencing revealed that vacuum-packaged samples contained a higher total bacterial richness (n = 69 operational taxonomic units [OTUs]) than samples under the other packaging condition (n = 46 OTUs). The core community was composed of 6 OTUs (Lactobacillus sakei, Lactococcus piscium, Carnobacterium divergens, Carnobacterium maltaromaticum, Serratia proteamaculans, and Brochothrix thermosphacta), whereas 13 OTUs taxonomically assigned to the Enterobacteriaceae, Enterococcaceae, and Leuconostocaceae families comprised a less-abundant subpopulation. This subdominant community was significantly more abundant when 2.0% salt and vacuum packaging were used, and this correlated with a lower degree of spoilage. Our results demonstrate that salt reduction, particularly when it is combined with CO2-enriched packaging, promotes faster spoilage of raw sausages by lowering the overall bacterial diversity (both richness and evenness). Our study takes place in the context of raw meat product manufacturing and is linked to a requirement for salt reduction. Health guidelines are calling for a reduction in dietary salt intake. However, salt has been used for a very long time as a hurdle technology, and salt reduction in meat products raises the question of spoilage and waste of food. The study was conceived to assess the role of sodium chloride reduction in meat products, both at the level of spoilage development and at the level of bacterial diversity, using 16S rRNA amplicon sequencing and raw pork sausage as a meat model. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Bio-protective potential of lactic acid bacteria: Effect of Lactobacillus sakei and Lactobacillus curvatus on changes of the microbial community in vacuum-packaged chilled beef.

PubMed

Zhang, Yimin; Zhu, Lixian; Dong, Pengcheng; Liang, Rongrong; Mao, Yanwei; Qiu, Shubing; Luo, Xin

2018-04-01

This study was to determine the bacterial diversity and monitor the community dynamic changes during storage of vacuum-packaged sliced raw beef as affected by Lactobacillus sakei and Lactobacillus curvatus . L. sakei and L. curvatus were separately incubated in vacuumed-packaged raw beef as bio-protective cultures to inhibit the naturally contaminating microbial load. Dynamic changes of the microbial diversity of inoculated or non-inoculated (control) samples were monitored at 4°C for 0 to 38 days, using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). The DGGE profiles of DNA directly extracted from non-inoculated control samples highlighted the order of appearance of spoilage bacteria during storage, showing that Enterbacteriaceae and Pseudomonas fragi emerged early, then Brochothrix thermosphacta shared the dominant position, and finally, Pseudomonas putida showed up became predominant. Compared with control, the inoculation of either L. sakei or L. curvatus significantly lowered the complexity of microbial diversity and inhibited the growth of spoilage bacteria (p<0.05). Interestingly, we also found that the dominant position of L. curvatus was replaced by indigenous L. sakei after 13 d for L. curvatus -inoculated samples. Plate counts on selective agars further showed that inoculation with L. sakei or L. curvatus obviously reduced the viable counts of Enterbacteraceae , Pseudomonas spp. and B. thermosphacta during later storage (p< 0.05), with L. sakei exerting greater inhibitory effect. Inoculation with both bio-protective cultures also significantly decreased the total volatile basic nitrogen values of stored samples (p<0.05). Taken together, the results proved the benefits of inoculation with lactic acid bacteria especially L. sakei as a potential way to inhibit growth of spoilage-related bacteria and improve the shelf life of vacuum-packaged raw beef.

Chemical and biochemical study of industrially produced San Simón da Costa smoked semi-hard cow's milk cheeses: Effects of storage under vacuum and different modified atmospheres.

PubMed

Garabal, J I; Rodríguez-Alonso, P; Franco, D; Centeno, J A

2010-05-01

Two batches of smoked, semi-hard (ripened for 45 d) San Simón da Costa cow's milk cheeses with Protected Designation of Origin were used to investigate the chemical, biochemical, and sensorial parameters that may be affected by modified-atmosphere packaging. Cheeses were packaged for 45 d as follows: vacuum packaging, packaging in 100% N(2), packaging in a gas mixture of 20% CO(2)/80% N(2), and packaging in a gas mixture of 50% CO(2)/50% N(2). The San Simón da Costa cheeses were characterized by high contents of lactic, oxalic, and citric organic acids. The main free amino acids found were isoleucine, phenylalanine, serine, valine, lysine, and glutamic acid, and the most abundant volatile compounds included ethanol, diacetyl, 2-butanol, isopropyl alcohol, furfural, acetaldehyde, 2-butanone, acetone, and 2-methylfuran. Modified atmospheres appeared to alter the ripening processes by affecting lipolysis, as indicated by the lower concentrations of butyric and propionic acids compared with control cheeses. In addition, modified-atmosphere packaging altered the proteolysis processes, yielding higher amounts of branched-chain alcohols. The results revealed that storage under modified atmosphere contributes to the accumulation of several compounds probably derived from smoke, including aldehydes such as 2-furancarboxaldehyde (furfural), alcohols such as 2-methoxyphenol (guaiacol), ketones such as 2-cyclopenten-1-one, and esters such as methyl furancarboxylate, which were negatively correlated with flavor. Vacuum packaging was the most useful technique in terms of preserving the sensory quality of San Simón da Costa Protected Designation of Origin cheeses. Considering the current demands for packaged portions of food at the distribution and retail levels and the potential health risks associated with some smoke-derived compounds usually present in some smoked foods, the results obtained in this study may be of special interest to the cheese industry. Copyright 2010 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Lactic acid bacteria of meat and meat products.

PubMed

Egan, A F

1983-09-01

When the growth of aerobic spoilage bacteria is inhibited, lactic acid bacteria may become the dominant component of the microbial flora of meats. This occurs with cured meats and with meats packaged in films of low gas permeability. The presence of a flora of psychrotrophic lactic acid bacteria on vacuum-packaged fresh chilled meats usually ensures that shelf-life is maximal. When these organisms spoil meats it is generally by causing souring, however other specific types of spoilage do occur. Some strains cause slime formation and greening of cured meats, and others may produce hydrogen sulphide during growth on vacuum-packaged beef. The safety and stability of fermented sausages depends upon fermentation caused by lactic acid bacteria. Overall the presence on meats of lactic acid bacteria is more desirable than that of the types of bacteria they have replaced.

Structural Characterization of H-1 Parvovirus: Comparison of Infectious Virions to Empty Capsids

PubMed Central

Halder, Sujata; Nam, Hyun-Joo; Govindasamy, Lakshmanan; Vogel, Michèle; Dinsart, Christiane; Salomé, Nathalie; McKenna, Robert

2013-01-01

The structure of single-stranded DNA (ssDNA) packaging H-1 parvovirus (H-1PV), which is being developed as an antitumor gene delivery vector, has been determined for wild-type (wt) virions and noninfectious (empty) capsids to 2.7- and 3.2-Å resolution, respectively, using X-ray crystallography. The capsid viral protein (VP) structure consists of an α-helix and an eight-stranded anti-parallel β-barrel with large loop regions between the strands. The β-barrel and loops form the capsid core and surface, respectively. In the wt structure, 600 nucleotides are ordered in an interior DNA binding pocket of the capsid. This accounts for ∼12% of the H-1PV genome. The wt structure is identical to the empty capsid structure, except for side chain conformation variations at the nucleotide binding pocket. Comparison of the H-1PV nucleotides to those observed in canine parvovirus and minute virus of mice, two members of the genus Parvovirus, showed both similarity in structure and analogous interactions. This observation suggests a functional role, such as in capsid stability and/or ssDNA genome recognition for encapsulation. The VP structure differs from those of other parvoviruses in surface loop regions that control receptor binding, tissue tropism, pathogenicity, and antibody recognition, including VP sequences reported to determine tumor cell tropism for oncotropic rodent parvoviruses. These structures of H-1PV provide insight into structural features that dictate capsid stabilization following genome packaging and three-dimensional information applicable for rational design of tumor-targeted recombinant gene delivery vectors. PMID:23449783

New class of microminiature Joule — Thomson refrigerator and vacuum package

NASA Astrophysics Data System (ADS)

Paugh, Robert L.

1990-12-01

Progress is reported on the development of a two-stage, fast cooldown Joule — Thomson refrigerator using nitrogen gas and a nitrogen — hydrocarbon gas mixture as the refrigerants. The refrigerator incorporates a microminiature Venturi pump to reduce the pressure of the exhaust of the main boiler to bring the operating temperature of the cold stage to < 70 K in as little as 10 s. The vacuum package for the refrigerator contains no organic materials and is designed to provide a ten year shelf life. Special glass strengthening techniques are being used to achieve cooler survival of acceleration tests of up to 100 000g.

The effectiveness of triclosan-incorporated plastic against bacteria on beef surfaces.

PubMed

Cutter, C N

1999-05-01

Triclosan is a nonionic, broad-spectrum, antimicrobial agent that has been incorporated into a variety of personal hygiene products, including hand soaps, deodorants, shower gels, mouthwashes, and toothpastes. In this study, plastic containing 1,500 ppm of triclosan was evaluated in plate overlay assays and meat experiments as a means of reducing populations of bacteria. Plate overlay assays indicated that the triclosan-incorporated plastic (TIP) inhibited the following organisms: Brochothrix thermosphacta ATCC 11509, Salmonella Typhimurium ATCC 14028, Staphylococcus aureus ATCC 12598, Bacillus subtilis ATCC 6051, Shigella flexneri ATCC 12022, Escherichia coli ATCC 25922, and several strains of E. coli O157:H7. In meat experiment 1, irradiated, lean beef surfaces inoculated with B. thermosphacta, Salmonella Typhimurium, E. coli O157:H7, or B. subtilis were covered with TIP, vacuum packaged, and stored for 24 h at 4 degrees C. Of the organisms tested, only populations of B. thermosphacta were slightly reduced. In meat experiment 2, prerigor beef surfaces were inoculated with E. coli O157: H7, Salmonella Typhimurium, or B. thermosphacta incubated at 4 degrees C for 24 h, wrapped in TIP or control plastic, vacuum packaged, and stored at 4 degrees C for up to 14 days. There was a slight reduction in the population of the organisms after initial application with TIP. However, bacterial populations following long-term, refrigerated (4 degrees C), vacuum-packaged storage up to 14 days were not statistically (P< or =0.05) or numerically different than controls. In meat experiment 3, even TIP-wrapped, vacuum-packaged beef samples that were temperature abused at 12 degrees C did not exhibit significant (P< or =0.05) or sustainable reductions after 14 days of 4 degrees C storage. Another study indicated that populations of E. coli O157:H7 or B. thermosphacta added directly to TIP were not affected after 2 h of refrigerated storage or that the antimicrobial activity could be extracted from the plastic. Additional experiments suggest that presence of fatty acids or adipose may diminish the antimicrobial activity of TIP on meat surfaces. This study demonstrates that while antimicrobial activity is detected against bacterial cultures in antimicrobial plate assays, plastic containing 1,500 ppm of triclosan does not effectively reduce bacterial populations on refrigerated, vacuum-packaged meat surfaces.

Transfer orbit stage mechanisms thermal vacuum test

NASA Technical Reports Server (NTRS)

Oleary, Scott T.

1990-01-01

A systems level mechanisms test was conducted on the Orbital Sciences Corp.'s Transfer Orbit Stage (TOS). The TOS is a unique partially reusable transfer vehicle which will boost a satellite into its operational orbit from the Space Shuttle's cargo bay. The mechanical cradle and tilt assemblies will return to earth with the Space Shuttle while the Solid Rocket Motor (SRM) and avionics package are expended. A mechanisms test was performed on the forward cradle and aft tilting assemblies of the TOS under thermal vacuum conditions. Actuating these assemblies under a 1 g environment and thermal vacuum conditions proved to be a complex task. Pneumatic test fixturing was used to lift the forward cradle, and tilt the SRM, and avionics package. Clinometers, linear voltage displacement transducers, and load cells were used in the thermal vacuum chamber to measure the performance and characteristics of the TOS mechanism assembly. Incorporation of the instrumentation and pneumatic system into the test setup was not routine since pneumatic actuation of flight hardware had not been previously performed in the facility. The methods used are presented along with the problems experienced during the design, setup and test phases.

Method and apparatus for in-cell vacuuming of radiologically contaminated materials

DOEpatents

Spadaro, Peter R.; Smith, Jay E.; Speer, Elmer L.; Cecconi, Arnold L.

1987-01-01

A vacuum air flow operated cyclone separator arrangement for collecting, handling and packaging loose contaminated material in accordance with acceptable radiological and criticality control requirements. The vacuum air flow system includes a specially designed fail-safe prefilter installed upstream of the vacuum air flow power supply. The fail-safe prefilter provides in-cell vacuum system flow visualization and automatically reduces or shuts off the vacuum air flow in the event of an upstream prefilter failure. The system is effective for collecting and handling highly contaminated radiological waste in the form of dust, dirt, fuel element fines, metal chips and similar loose material in accordance with radiological and criticality control requirements for disposal by means of shipment and burial.

Silicone-oil-based subvisible particles: their detection, interactions, and regulation in prefilled container closure systems for biopharmaceuticals.

PubMed

Felsovalyi, Flora; Janvier, Sébastien; Jouffray, Sébastien; Soukiassian, Hervé; Mangiagalli, Paolo

2012-12-01

Recent increased regulatory scrutiny concerning subvisible particulates (SbVPs) in parenteral formulations of biologics has led to the publication of numerous articles about the sources, characteristics, implications, and approaches to monitoring and detecting SbVPs. Despite varying opinions on the level of associated risks and method of regulation, nearly all industry scientists and regulators agree on the need for monitoring and reporting visible and subvisible particles. As prefillable drug delivery systems have become a prominent packaging option, silicone oil, a common primary packaging lubricant, may play a role in the appearance of particles. The goal of this article is to complement the current SbVP knowledge base with new insights into the evolution of silicone-oil-related particulates and their interactions with components in prefillable systems. We propose a "toolbox" for improved silicone-oil-related particulate detection and enumeration, and discuss the benefits and limitations of approaches for lowering and controlling silicone oil release in parenterals. Finally, we present surface cross-linking of silicone as the recommended solution for achieving significant SbVP reduction without negatively affecting functional performance. Copyright © 2012 Wiley Periodicals, Inc.

Interaction and interdependent packaging of tegument protein UL11 and glycoprotein e of herpes simplex virus.

PubMed

Han, Jun; Chadha, Pooja; Meckes, David G; Baird, Nicholas L; Wills, John W

2011-09-01

The UL11 tegument protein of herpes simplex virus plays a critical role in the secondary envelopment; however, the mechanistic details remain elusive. Here, we report a new function of UL11 in the budding process in which it directs efficient acquisition of glycoprotein E (gE) via a direct interaction. In vitro binding assays showed that the interaction required only the first 28, membrane-proximal residues of the cytoplasmic tail of gE, and the C-terminal 26 residues of UL11. A second, weaker binding site was also found in the N-terminal half of UL11. The significance of the gE-UL11 interaction was subsequently investigated with viral deletion mutants. In the absence of the gE tail, virion packaging of UL11, but not other tegument proteins such as VP22 and VP16, was reduced by at least 80%. Reciprocally, wild-type gE packaging was also drastically reduced by about 87% in the absence of UL11, and this defect could be rescued in trans by expressing U(L)11 at the U(L)35 locus. Surprisingly, a mutant that lacks the C-terminal gE-binding site of UL11 packaged nearly normal amounts of gE despite its strong interaction with the gE tail in vitro, indicating that the interaction with the UL11 N terminus may be important. Mutagenesis studies of the UL11 N terminus revealed that the association of UL11 with membrane was not required for this function. In contrast, the UL11 acidic cluster motif was found to be critical for gE packaging and was not replaceable with foreign acidic clusters. Together, these results highlight an important role of UL11 in the acquisition of glycoprotein-enriched lipid bilayers, and the findings may also have important implications for the role of UL11 in gE-mediated cell-to-cell spread.

Variability of High Resolution Vp/Vs and Seismic Velocity Structure Along the Nicaragua/Costa Rica Segment of the Middle America Subduction Zone

NASA Astrophysics Data System (ADS)

Moore-Driskell, M. M.; DeShon, H. R.

2012-12-01

Previous studies of subduction zone earthquakes have shown that fault conditions control earthquake rupture and behavior. There are many potential properties that may vary along the subduction margin that could cause fault zone variability, including plate age, temperature, and/or geometry, convergence rate, state of hydration, overriding geology, subducting sediment packages, or subducting seamounts/ridges. The Nicaragua/Costa Rica segment of the Middle America subduction zone is highly variable along strike and down dip. We use this margin to examine how these variable conditions affect earthquake behavior by determining local ratios of compressional to shear wave velocities (Vp/Vs) and detailed seismic velocity structure. Vp/Vs is one of the best tools available to reliably define fault conditions because it is directly related to the Poisson's ratio of the fault material, and it is sensitive to the presence of fluids and changing permeability. Thus with well-resolved near source Vp/Vs measurements we can infer composition and/or high fluid pressures. Here, we use a technique developed by Lin and Shearer (2007) to determine local Vp/Vs in small areas (~2 x 2 x 2 km) with high seismicity. Within the seismogenic zone, we find the margin to be highly variable along strike in Vp/Vs and seismic velocity. These changes correlate to documented variability in incoming plate properties. Increased Vp/Vs is associated with intraplate earthquakes along Nicaragua and northern Costa Rica. We compare our results with other geophysical studies including new high-resolution images of seismic velocity structure, an extensive catalog of high quality relocated events, apparent stress calculations, coupling, and SSE/NVT occurrence. A better understanding of the connection between fault properties and earthquake behavior gives insight into the role of fluids in seismogenesis, the spectrum of earthquake rupture, and possible hazard at subduction zones.

The effect of heat shrink treatment and storage temperature on the time of onset of "blown pack" spoilage.

PubMed

Moschonas, Galatios; Bolton, Declan J; Sheridan, James J; McDowell, David A

2011-02-01

This study determined the effects of (a) the short "heat shrink" treatment frequently applied to vacuum packed meats within normal commercial production, and (b) chill holding storage temperature, on the subsequent time to onset (TTO) of "blown pack" spoilage (BPS). Beef or lamb steaks were inoculated with 10³ CFU/cm² of spore suspensions of five gas producing clostridia, vacuum packed (VP) and treated as follows: no heat, 50°C/15 s, 70°C/10 s or 90°C/3 s. Samples were stored at -1.5, 1 or 4°C and examined daily to determine TTO of BPS. For each strain, pack treatment and storage temperature had significant (P<0.05 and P<0.001 respectively) effects on TTO of BPS, i.e. 90°C/3 s<70°C/10 s<50°C/15 s≤"no heat", and 4°C<1°C<-1.5°C. The study suggested that the meat industry could reduce the risks of BPS by avoiding higher temperature (90°C/3 s or 70°C/10 s) heat shrinking, and by storing VP meats at lower temperatures (e.g. -1.5°C). Copyright © 2010 The American Meat Science Association. Published by Elsevier Ltd. All rights reserved.

Packaging Concerns and Techniques for Large Devices: Challenges for Complex Electronics

NASA Technical Reports Server (NTRS)

LaBel, Kenneth A.; Sampson, Michael J.

2010-01-01

NASA is going to have to accept the use of non-hermetic packages for complex devices. There are a large number of packaging options available. Space application subjects the packages to stresses that they were probably not designed for (vacuum for instance). NASA has to find a way of having assurance in the integrity of the packages. There are manufacturers interested in qualifying non-hermetic packages to MIL-PRF-38535 Class V. Government space users are agreed that Class V should be for hermetic packages only. NASA is working on a new Class for non-hermetic packages for M38535 Appendix B, "Class Y". Testing for package integrity will be required but can be package specific as described by a Package Integrity Test Plan. The plan is developed by the manufacturer and approved by DSCC and government space.

High-sensitivity Leak-testing Method with High-Resolution Integration Technique

NASA Astrophysics Data System (ADS)

Fujiyoshi, Motohiro; Nonomura, Yutaka; Senda, Hidemi

A high-resolution leak-testing method named HR (High-Resolution) Integration Technique has been developed for MEMS (Micro Electro Mechanical Systems) sensors such as a vibrating angular-rate sensor housed in a vacuum package. Procedures of the method to obtain high leak-rate resolution were as follows. A package filled with helium gas was kept in a small accumulation chamber to accumulate helium gas leaking from the package. After the accumulation, the accumulated helium gas was introduced into a mass spectrometer in a short period of time, and the flux of the helium gas was measured by the mass spectrometer as a transient phenomenon. The leak-rate of the package was calculated from the detected transient waveform of the mass spectrometer and the accumulation time of the helium gas in the accumulation chamber. Because the density of the helium gas in the vacuum chamber increased and the accumulated helium gas was measured in a very short period of time with the mass spectrometer, the peak strength of the transient waveform became high and the signal to noise ratio was much improved. The detectable leak-rate resolution of the technique reached 1×10-15 (Pa·m3/s). This resolution is 103 times superior to that of the conventional helium vacuum integration method. The accuracy of the measuring system was verified with a standard helium gas leak source. The results were well matched between theoretical calculation based on the leak-rate of the source and the experimental results within only 2% error.

Colour, lipid and protein stability of Rhea americana meat during air- and vacuum-packaged storage: influence of muscle on oxidative processes.

PubMed

Filgueras, R S; Gatellier, P; Aubry, L; Thomas, A; Bauchart, D; Durand, D; Zambiazi, R C; Santé-Lhoutellier, V

2010-11-01

Physicochemical characteristics and oxidative stability during storage were determined in Gastrocnemius pars interna (GN) and Iliofiburalis (IF) muscles of Rhea americana. Glycolytic potential (GP) and pH decline of muscles were measured within the first 24 h post mortem. Colour, lipid and protein stability were determined during storage of meat, i.e. 5 days under air-packaging at 4°C, or 28 days under vacuum-packaging at 4°C. In parallel, anti-oxidant status of muscles was estimated by measuring α-tocopherol content and anti-oxidant enzyme activities (superoxide dismutase and catalase), while pro-oxidant status was evaluated by determining haeminic iron and long chain fatty acids (especially polyunsaturated fatty acids). The ultimate pH was similar in both muscles, but the GP value was significantly higher in IF than in GN muscle. Haeminic iron and alpha-tocopherol content differed between muscles, with 30% more haeminic iron (p<0.05) and 134% more alpha-tocopherol (p<0.001) in IF than GN muscle. The IF muscle presented higher lipid content and lower PUFA/SFA ratio (polyunsaturated fatty acids/saturated fatty acids) than GN muscle. With storage under air-packaging, lipid and protein oxidation of rhea muscles increased up to 275% and 30%, respectively. This increase was more rapidly and marked in IF muscle. The IF also showed high level of metmyoglobin accumulation after 3 days of storage (47%) and was rejected by 1 consumer out of 2 in sensorial analysis. Under vacuum-packaging, both muscles showed a high stability of colour and no oxidation of lipids and proteins. Copyright © 2010 The American Meat Science Association. Published by Elsevier Ltd. All rights reserved.

7 CFR 319.73-3 - Conditions for transit movement of certain products through Puerto Rico or Hawaii.

Code of Federal Regulations, 2012 CFR

2012-01-01

... Agriculture (Continued) ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE FOREIGN... not limited to, sealed cartons, airtight containers, or vacuum packaging. Samples of unroasted coffee received by mail but not packaged in this manner are subject to inspection and safeguard by an inspector...

7 CFR 319.73-3 - Conditions for transit movement of certain products through Puerto Rico or Hawaii.

Code of Federal Regulations, 2011 CFR

2011-01-01

... Agriculture (Continued) ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE FOREIGN... not limited to, sealed cartons, airtight containers, or vacuum packaging. Samples of unroasted coffee received by mail but not packaged in this manner are subject to inspection and safeguard by an inspector...

7 CFR 319.73-3 - Conditions for transit movement of certain products through Puerto Rico or Hawaii.

Code of Federal Regulations, 2014 CFR

2014-01-01

... Agriculture (Continued) ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE FOREIGN... not limited to, sealed cartons, airtight containers, or vacuum packaging. Samples of unroasted coffee received by mail but not packaged in this manner are subject to inspection and safeguard by an inspector...

7 CFR 319.73-3 - Conditions for transit movement of certain products through Puerto Rico or Hawaii.

Code of Federal Regulations, 2010 CFR

2010-01-01

... Agriculture (Continued) ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE FOREIGN... not limited to, sealed cartons, airtight containers, or vacuum packaging. Samples of unroasted coffee received by mail but not packaged in this manner are subject to inspection and safeguard by an inspector...

7 CFR 319.73-3 - Conditions for transit movement of certain products through Puerto Rico or Hawaii.

Code of Federal Regulations, 2013 CFR

2013-01-01

... Agriculture (Continued) ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE FOREIGN... not limited to, sealed cartons, airtight containers, or vacuum packaging. Samples of unroasted coffee received by mail but not packaged in this manner are subject to inspection and safeguard by an inspector...

Lipid oxidation and color changes of goose meat stored under vacuum and modified atmosphere conditions.

PubMed

Orkusz, A; Haraf, G; Okruszek, A; Werenska-Sudnik, M

2017-03-01

The objective of the work was to investigate the color and lipid oxidation changes of goose breast meat packaged in vacuum and modified atmosphere (MA) conditions consisting of 80% O2, 20% CO2, and stored in refrigerated conditions at 4°C. Color stability was monitored by determining total heme pigments concentration; relative concentration of myoglobin, oxymyoglobin, and metmyoglobin; parameters of color L*, a*, b*, and sensory evaluation of the surface color. Lipid stability was measured by determining thiobarbituric acid reactive substances (TBARS). The samples were examined in 24 h after slaughter (unpacked muscles) and on d 4, 7, 9, 11 of storage (muscles packed in vacuum and in MA). Through the time of storage, samples packed in MA had higher TBARS values in comparison to the meat packed in vacuum. For samples packed in two types of atmospheres, the total pigments concentration decreased gradually within 11 d of storage. It was observed that relative metmyoglobin concentration increased whereas relative oxymyoglobin concentration decreased in total heme pigments in the MA stored muscle. The relative concentration of all three myoglobin forms sample packed in vacuum remained unchanged. The color parameters (L*, a*, b*) did not change for 11 d of storage for the vacuum packed meat. The value of the color parameter a* decreased and the value of the color parameters L* and b* increased in the samples packaged in MA. The data prove that if you store goose meat in MA (consisting of 80% O2, 20% CO2) or vacuum, the unchanged surface color is preserved for 9 and 11 day, respectively.Vacuum appears to be a better method as regards the maintaining of lipid stability in goose meat. © 2016 Poultry Science Association Inc.

Dissolved carbon dioxide and oxygen concentrations in purge of vacuum-packaged pork chops and the relationship to shelf life and models for estimating microbial populations.

PubMed

Adams, K R; Niebuhr, S E; Dickson, J S

2015-12-01

The objectives of this study were to determine the dissolved CO2 and O2 concentrations in the purge of vacuum-packaged pork chops over a 60 day storage period, and to elucidate the relationship of dissolved CO2 and O2 to the microbial populations and shelf life. As the populations of spoilage bacteria increased, the dissolved CO2 increased and the dissolved O2 decreased in the purge. Lactic acid bacteria dominated the spoilage microflora, followed by Enterobacteriaceae and Brochothrix thermosphacta. The surface pH decreased to 5.4 due to carbonic acid and lactic acid production before rising to 5.7 due to ammonia production. A mathematical model was developed which estimated microbial populations based on dissolved CO2 concentrations. Scanning electron microscope images were also taken of the packaging film to observe the biofilm development. The SEM images revealed a two-layer biofilm on the packaging film that was the result of the tri-phase growth environment. Copyright © 2015 Elsevier Ltd. All rights reserved.

Effect of packaging type during postmortem aging and degree of doneness on pork chop sensory traits of loins selected to vary in color and marbling.

PubMed

Klehm, B J; King, D A; Dilger, A C; Shackelford, S D; Boler, D D

2018-05-04

The objective was to determine the interactions between packaging type and degree of doneness on sensory traits of pork loins classified based on the newly proposed USDA quality grades. A total of 144 loins were selected from 2 groups of pigs (lean growth or meat quality production focus) to represent as much variation in visual color and marbling as possible. Selection was achieved with a VQG grading camera. The ventral surface of the loins was evaluated for loin quality traits at 1 d postmortem. At 2 d postmortem loins were sliced into 28-mm-thick chops. Chop within each loin was randomly assigned to either individual vacuum packages or to individual Styrofoam trays and overwrapped in polyvinyl chloride (PVC) oxygen permeable film. Overwrapped PVC packages were then placed in bulk packages and flushed with a gas mixture that contained approximately 0.4% carbon monoxide, 30% carbon dioxide, and 80% nitrogen. Vacuum-packaged chops were aged until 14 d postmortem. Chops packaged in PVC overwrap were aged until 9 d postmortem in the bulk packages, then placed on simulated retail display until 14 d postmortem. Chops from each packaging type were cooked to an internal temperature of either 63 °C or 71 °C for the evaluation of slice shear force (SSF) or for evaluation of tenderness, juiciness, and flavor by a trained panel. Data were analyzed as split-split plot design with production focus of the pigs, proposed USDA quality grade, packaging type, and degree of doneness as fixed effects. While there were main effect differences between production focuses, there were no interactions with production focus. There were also no 3-way (P ≥ 0.19) interactions and only one 2-way interaction among quality grade, packaging type, or degree of doneness. There were no differences in sensory tenderness (P = 0.30), juiciness (P = 0.49), flavor (P = 0.89), SSF (P = 0.13), or cook loss (P = 0.06) among USDA quality grades. There were no differences in sensory tenderness (P = 0.06), juiciness (P = 0.32), flavor (P = 0.74), SSF (P = 0.99), or cook loss (P = 0.12) between chops aged in vacuum packages or PVC packages. Chops cooked to 63 °C were 4.6% more tender (P < 0.0001), 10.1% juicier (P < 0.0001), and 2.9% less flavorful (P = 0.01) than chops cooked to 71 °C. These data suggest that cooking chops to 63 °C rather than 71 °C was a more effective way to improve tenderness and juiciness than selecting chops of a certain quality grade or altering packaging postmortem.

Cryo-Electron Tomography of Marburg Virus Particles and Their Morphogenesis within Infected Cells

PubMed Central

Kolesnikova, Larissa; Welsch, Sonja; Krähling, Verena; Davey, Norman; Parsy, Marie-Laure; Becker, Stephan; Briggs, John A. G.

2011-01-01

Several major human pathogens, including the filoviruses, paramyxoviruses, and rhabdoviruses, package their single-stranded RNA genomes within helical nucleocapsids, which bud through the plasma membrane of the infected cell to release enveloped virions. The virions are often heterogeneous in shape, which makes it difficult to study their structure and assembly mechanisms. We have applied cryo-electron tomography and sub-tomogram averaging methods to derive structures of Marburg virus, a highly pathogenic filovirus, both after release and during assembly within infected cells. The data demonstrate the potential of cryo-electron tomography methods to derive detailed structural information for intermediate steps in biological pathways within intact cells. We describe the location and arrangement of the viral proteins within the virion. We show that the N-terminal domain of the nucleoprotein contains the minimal assembly determinants for a helical nucleocapsid with variable number of proteins per turn. Lobes protruding from alternate interfaces between each nucleoprotein are formed by the C-terminal domain of the nucleoprotein, together with viral proteins VP24 and VP35. Each nucleoprotein packages six RNA bases. The nucleocapsid interacts in an unusual, flexible “Velcro-like” manner with the viral matrix protein VP40. Determination of the structures of assembly intermediates showed that the nucleocapsid has a defined orientation during transport and budding. Together the data show striking architectural homology between the nucleocapsid helix of rhabdoviruses and filoviruses, but unexpected, fundamental differences in the mechanisms by which the nucleocapsids are then assembled together with matrix proteins and initiate membrane envelopment to release infectious virions, suggesting that the viruses have evolved different solutions to these conserved assembly steps. PMID:22110401

A low feed-through 3D vacuum packaging technique with silicon vias for RF MEMS resonators

NASA Astrophysics Data System (ADS)

Zhao, Jicong; Yuan, Quan; Kan, Xiao; Yang, Jinling; Yang, Fuhua

2017-01-01

This paper presents a wafer-level three-dimensional (3D) vacuum packaging technique for radio frequency microelectromechanical systems (RF MEMS) resonators. A Sn-rich Au-Sn solder bonding is employed to provide a vacuum encapsulation as well as electrical conductions. Vertical silicon vias are micro-fabricated by glass reflow process. The optimized grounding, via pitch, and all-round shielding effectively reduce feed-through capacitance. Thus the signal-to-background ratios (SBRs) of the transmission signals increase from 17 dB to 20 dB, and the quality factor (Q) values of the packaged resonators go from around 8000 up to more than 9500. The measured average leak rate and shear strength are (2.55  ±  0.9)  ×  10-8 atm-cc s-1 and 42.53  ±  4.19 MPa, respectively. Furthermore, thermal cycling test between  -40 °C and 100 °C and high temperature storage test at 150 °C show that the resonant-frequency drifts are less than  ±7 ppm. In addition, the SBRs and the Q values have no obvious change after the tests. The experimental results demonstrated that the proposed encapsulation technique is well suited for the applications of RF MEMS devices.

Screening of lactic acid bacteria from vacuum packaged beef for antimicrobial activity

PubMed Central

Oliveira, Roseane B. P.; de L. Oliveira, Afonso; Glória, M. Beatriz A.

2008-01-01

The objective of this study was to isolate lactic acid bacteria (LAB) from vacuum packaged beef and to investigate their antagonist activity. LAB mean counts of 5.19 log cfu/cm2 were obtained from five samples of vacuum packaged beef. Two hundred isolates were selected and screened for the inhibitory effect on five ATCC reference Lactobacillus strains. Thirty six isolates showed activity in the agar spot test against at least two of the indicator strains. However, only six cell free supernatants (CFS) from these isolates exhibited activity against the indicator strains using the well-diffusion test and conditions that eliminated the effects of organic acids and hydrogen peroxide. L. acidophilus was the most sensitive indicator tested, whereas L. plantarum and L. fermentum were the most resistant ones. Identification by MIDI system indicated that these LAB isolates were Lactococcus lactis subsp. cremoris, Pediococcus acidilactici, Lactobacillus delbrueckii subsp. bulgaricus and Lactobacillus casei GC subgroup A. The antagonistic factors produced by most of these LAB against L. acidophilus were resistant to heat treatment (100°C for 10 min) and stable over a wide pH range (4.0 to 9.0). These data suggest that these isolates could be used as promising hurdles aiming increased safety and extended shelf life of meat products. PMID:24031232

Bio-protective potential of lactic acid bacteria: Effect of Lactobacillus sakei and Lactobacillus curvatus on changes of the microbial community in vacuum-packaged chilled beef

PubMed Central

Zhang, Yimin; Zhu, Lixian; Dong, Pengcheng; Liang, Rongrong; Mao, Yanwei; Qiu, Shubing

2018-01-01

Objective This study was to determine the bacterial diversity and monitor the community dynamic changes during storage of vacuum-packaged sliced raw beef as affected by Lactobacillus sakei and Lactobacillus curvatus. Methods L. sakei and L. curvatus were separately incubated in vacuumed-packaged raw beef as bio-protective cultures to inhibit the naturally contaminating microbial load. Dynamic changes of the microbial diversity of inoculated or non-inoculated (control) samples were monitored at 4°C for 0 to 38 days, using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Results The DGGE profiles of DNA directly extracted from non-inoculated control samples highlighted the order of appearance of spoilage bacteria during storage, showing that Enterbacteriaceae and Pseudomonas fragi emerged early, then Brochothrix thermosphacta shared the dominant position, and finally, Pseudomonas putida showed up became predominant. Compared with control, the inoculation of either L. sakei or L. curvatus significantly lowered the complexity of microbial diversity and inhibited the growth of spoilage bacteria (p<0.05). Interestingly, we also found that the dominant position of L. curvatus was replaced by indigenous L. sakei after 13 d for L. curvatus-inoculated samples. Plate counts on selective agars further showed that inoculation with L. sakei or L. curvatus obviously reduced the viable counts of Enterbacteraceae, Pseudomonas spp. and B. thermosphacta during later storage (p< 0.05), with L. sakei exerting greater inhibitory effect. Inoculation with both bio-protective cultures also significantly decreased the total volatile basic nitrogen values of stored samples (p<0.05). Conclusion Taken together, the results proved the benefits of inoculation with lactic acid bacteria especially L. sakei as a potential way to inhibit growth of spoilage-related bacteria and improve the shelf life of vacuum-packaged raw beef. PMID:29059725

[Impact of storage conditions and time on herb of Lonicera macranthoides].

PubMed

Ma, Peng; Li, Long-Yun; Zhang, Ying

2014-03-01

To study the effect of different storage conditions and storage time on herb quality of Lonicera macranthoides, different packaging materials including vacuum plastic bags, plastic bags, woven bags, sealed with endometrial bags, paper bags, sack bags were selected for the study under different storage conditions including room temperature, 5 degrees C refrigerator, low temperature of - 20 degrees C refrigerator and desiccator. Twenty-four batches of samples were used for the study, and active ingredients were determined. The experimental results showed that the ingredients in each storage group changed with the storage time, storage conditions (storage environment, packaging). Under the same storage time, the storage environment (temperature, humidity) had effect on the stability of herb quality. Low temperature had less effect on herb quality. The effect of packaging on herb quality was as following: plastic vacuum packaging > woven with endometrial sealed packaging > plastic bag > woven bag > sack bags > paper bags. Under the same storage conditions, with the increase of storage time, caffeic acid content increased slowly, and other five ingredients content decreased gradually. Storage time affected significantly on the intrinsic quality (chemical composition) and appearance of herb. It is suggested that low temperature (5 degrees C), dark and sealed storage are suitable for storage of L. macranthoides herb, the storage time should be not more than 24 months.

Development, fabrication and testing of a magnetically connected plastic vacuum probe surface sampler

NASA Technical Reports Server (NTRS)

Phillips, G. B.; Pace, V. A., Jr.

1972-01-01

The sampler utilizes permanent magnets and soft metal pole pieces to connect the cone/filter assembly to the sampling head and vacuum supply. The cone/filter assembly is packaged in a plastic container and presterilized so that the need for any human contact during the sampling procedure is completely eliminated. Microbiological tests have demonstrated that the sampling efficiency is not affected by the magnetic coupling apparatus and that the probe appears to function as efficiently as the conventional plastic and Sandia vacuum probes.

Use of Optical Oxygen Sensors in Non-Destructively Determining the Levels of Oxygen Present in Combined Vacuum and Modified Atmosphere Packaged Pre-Cooked Convenience-Style Foods and the Use of Ethanol Emitters to Extend Product Shelf-Life.

PubMed

Hempel, Andreas W; Papkovsky, Dmitri B; Kerry, Joseph P

2013-11-18

O₂ sensors were used to non-destructively monitor O₂ levels in commercially packed pre-cooked, convenience modified atmosphere packaging (MAP) foods. A substantial level of O₂ (>15%) was present in packs resulting in a shorter than expected shelf-life, where the primary spoilage mechanism was found to be mould. Various combinations of vacuum (0-0.6 MPa) and gas flush (0.02-0.03 MPa) (30% CO₂/70% N₂) settings were assessed as treatments that result in the desired shelf-life (28 days). This was achieved using the combined treatment of vacuum 0.35 MPa and gas flush 0.02 MPa which resulted in a reduction of 6%-9% O 2 in all three samples (battered sausages (BS), bacon slices (BA), and meat and potato pies (PP)). Reduced O₂ levels reflect the microbial quality of products, which has been successfully reduced. Duplicate samples of all product packs were produced using ethanol emitters (EE) to see if shelf-life could be further extended. Results showed a further improvement in shelf-life to 35 days. Sensory analysis showed that ethanol flavour and aroma was not perceived by panellists in two of the three products assessed. This study demonstrates how smart packaging technologies, both intelligent and active, can be used to assist in the modification of conventional packaging systems in order to enhance product quality and safety and through the extension of product shelf-life.

The Function of Emulsions on the Biogenic Amine Formation and their Indices of Sea Bass Fillets (Dicentrarchus Labrax) Stored in Vacuum Packaging.

PubMed

Ozogul, Yesim; Durmus, Mustafa; Kuley Boga, Esmeray; Uçar, Yılmaz; Ozogul, Fatih

2018-02-01

The impacts of emulsions based on commercial oils on the biogenic amine formation and their indices of vacuumed packed sea bass fillets were investigated. The results showed that among biogenic amines, cadaverine, putrescine, spermidine, spermine, serotonin, dopamine, and agmatine were predominant amines in sea bass fillets stored under vacuum packaging. Significant differences (P < 0.05) in biogenic amines concentrations of vacuumed packed sea bass treated with emulsions were observed. All groups contained histamine lower than 5.0 mg/100 g, regarded as the allowable limit by the U.S. Food and Drug Administration. Polyamine levels were not affected by application of emulsion. Quality index (QI) showed an increase and after 14 d of storage it decreased in all groups. The control generally seemed to higher QI value than those of treatment groups except at 14 and 18 days while soybean and corn gave lower QI among treatment groups. Only biogenic amine index correlated with sensory acceptability of vacuumed packed sea bass, indicating that this index can be used for determination of the degree of spoilage of vacuumed packed sea bass. Emulsions extended the shelf-life (approximately 2 to 4 d) of vacuumed packed sea bass fillets by inhibiting microbial growth compared to the control. Emulsions have become popular since they are regarded as ideal carrier for the delivery of lipophilic substances due to the ease of preparation, small particle size, their enhanced bioavailability, and long term kinetic stability. They have been proven to be self-preserving antimicrobials due to bound water in their structure and thus no available water to microorganisms. Antimicrobial emulsions have potential applications in many fields because they are inexpensive, stable, and nontoxic agents. © 2018 Institute of Food Technologists®.

Apparatus and method for skin packaging articles

NASA Technical Reports Server (NTRS)

Madsen, B.; Pozsony, E. R.; Collin, E. E. (Inventor)

1973-01-01

A system for skin packaging articles including a loading zone for positioning articles to be packaged upon a substrate, a thermoplastic film heating and vacuum operated skin packaging zone for covering the articles with film laminated to the substrate and a slitting zone for separating and trimming the individual skin packaged articles. The articles are passed to the successive zones. The loading zone may be adapted for conveyorized instead of hand loading. In some cases, where only transverse cutting of the film web is necessary, it may be desirable to eliminate the slitting zone and remove the skin packaged article or articles directly from the packaging zone. A conveniently located operating panel contains controls for effecting automatic, semiautomatic or manual operation of the entire system of any portions in any manner desired.

Structural Studies of Adeno-Associated Virus Serotype 8 Capsid Transitions Associated with Endosomal Trafficking

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

Nam, Hyun-Joo; Gurda, Brittney L.; McKenna, Robert

2012-09-17

The single-stranded DNA (ssDNA) parvoviruses enter host cells through receptor-mediated endocytosis, and infection depends on processing in the early to late endosome as well as in the lysosome prior to nuclear entry for replication. However, the mechanisms of capsid endosomal processing, including the effects of low pH, are poorly understood. To gain insight into the structural transitions required for this essential step in infection, the crystal structures of empty and green fluorescent protein (GFP) gene-packaged adeno-associated virus serotype 8 (AAV8) have been determined at pH values of 6.0, 5.5, and 4.0 and then at pH 7.5 after incubation at pHmore » 4.0, mimicking the conditions encountered during endocytic trafficking. While the capsid viral protein (VP) topologies of all the structures were similar, significant amino acid side chain conformational rearrangements were observed on (i) the interior surface of the capsid under the icosahedral 3-fold axis near ordered nucleic acid density that was lost concomitant with the conformational change as pH was reduced and (ii) the exterior capsid surface close to the icosahedral 2-fold depression. The 3-fold change is consistent with DNA release from an ordering interaction on the inside surface of the capsid at low pH values and suggests transitions that likely trigger the capsid for genome uncoating. The surface change results in disruption of VP-VP interface interactions and a decrease in buried surface area between VP monomers. This disruption points to capsid destabilization which may (i) release VP1 amino acids for its phospholipase A2 function for endosomal escape and nuclear localization signals for nuclear targeting and (ii) trigger genome uncoating.« less

Spatiotemporal Phylogenetic Analysis and Molecular Characterisation of Infectious Bursal Disease Viruses Based on the VP2 Hyper-Variable Region

PubMed Central

Dolz, Roser; Valle, Rosa; Perera, Carmen L.; Bertran, Kateri; Frías, Maria T.; Majó, Natàlia; Ganges, Llilianne; Pérez, Lester J.

2013-01-01

Background Infectious bursal disease is a highly contagious and acute viral disease caused by the infectious bursal disease virus (IBDV); it affects all major poultry producing areas of the world. The current study was designed to rigorously measure the global phylogeographic dynamics of IBDV strains to gain insight into viral population expansion as well as the emergence, spread and pattern of the geographical structure of very virulent IBDV (vvIBDV) strains. Methodology/Principal Findings Sequences of the hyper-variable region of the VP2 (HVR-VP2) gene from IBDV strains isolated from diverse geographic locations were obtained from the GenBank database; Cuban sequences were obtained in the current work. All sequences were analysed by Bayesian phylogeographic analysis, implemented in the Bayesian Evolutionary Analysis Sampling Trees (BEAST), Bayesian Tip-association Significance testing (BaTS) and Spatial Phylogenetic Reconstruction of Evolutionary Dynamics (SPREAD) software packages. Selection pressure on the HVR-VP2 was also assessed. The phylogeographic association-trait analysis showed that viruses sampled from individual countries tend to cluster together, suggesting a geographic pattern for IBDV strains. Spatial analysis from this study revealed that strains carrying sequences that were linked to increased virulence of IBDV appeared in Iran in 1981 and spread to Western Europe (Belgium) in 1987, Africa (Egypt) around 1990, East Asia (China and Japan) in 1993, the Caribbean Region (Cuba) by 1995 and South America (Brazil) around 2000. Selection pressure analysis showed that several codons in the HVR-VP2 region were under purifying selection. Conclusions/Significance To our knowledge, this work is the first study applying the Bayesian phylogeographic reconstruction approach to analyse the emergence and spread of vvIBDV strains worldwide. PMID:23805195

Spatiotemporal Phylogenetic Analysis and Molecular Characterisation of Infectious Bursal Disease Viruses Based on the VP2 Hyper-Variable Region.

PubMed

Alfonso-Morales, Abdulahi; Martínez-Pérez, Orlando; Dolz, Roser; Valle, Rosa; Perera, Carmen L; Bertran, Kateri; Frías, Maria T; Majó, Natàlia; Ganges, Llilianne; Pérez, Lester J

2013-01-01

Infectious bursal disease is a highly contagious and acute viral disease caused by the infectious bursal disease virus (IBDV); it affects all major poultry producing areas of the world. The current study was designed to rigorously measure the global phylogeographic dynamics of IBDV strains to gain insight into viral population expansion as well as the emergence, spread and pattern of the geographical structure of very virulent IBDV (vvIBDV) strains. Sequences of the hyper-variable region of the VP2 (HVR-VP2) gene from IBDV strains isolated from diverse geographic locations were obtained from the GenBank database; Cuban sequences were obtained in the current work. All sequences were analysed by Bayesian phylogeographic analysis, implemented in the Bayesian Evolutionary Analysis Sampling Trees (BEAST), Bayesian Tip-association Significance testing (BaTS) and Spatial Phylogenetic Reconstruction of Evolutionary Dynamics (SPREAD) software packages. Selection pressure on the HVR-VP2 was also assessed. The phylogeographic association-trait analysis showed that viruses sampled from individual countries tend to cluster together, suggesting a geographic pattern for IBDV strains. Spatial analysis from this study revealed that strains carrying sequences that were linked to increased virulence of IBDV appeared in Iran in 1981 and spread to Western Europe (Belgium) in 1987, Africa (Egypt) around 1990, East Asia (China and Japan) in 1993, the Caribbean Region (Cuba) by 1995 and South America (Brazil) around 2000. Selection pressure analysis showed that several codons in the HVR-VP2 region were under purifying selection. To our knowledge, this work is the first study applying the Bayesian phylogeographic reconstruction approach to analyse the emergence and spread of vvIBDV strains worldwide.

Event visualization in ATLAS

NASA Astrophysics Data System (ADS)

Bianchi, R. M.; Boudreau, J.; Konstantinidis, N.; Martyniuk, A. C.; Moyse, E.; Thomas, J.; Waugh, B. M.; Yallup, D. P.; ATLAS Collaboration

2017-10-01

At the beginning, HEP experiments made use of photographical images both to record and store experimental data and to illustrate their findings. Then the experiments evolved and needed to find ways to visualize their data. With the availability of computer graphics, software packages to display event data and the detector geometry started to be developed. Here, an overview of the usage of event display tools in HEP is presented. Then the case of the ATLAS experiment is considered in more detail and two widely used event display packages are presented, Atlantis and VP1, focusing on the software technologies they employ, as well as their strengths, differences and their usage in the experiment: from physics analysis to detector development, and from online monitoring to outreach and communication. Towards the end, the other ATLAS visualization tools will be briefly presented as well. Future development plans and improvements in the ATLAS event display packages will also be discussed.

Influence of modified atmospheric storage, lactic acid, and NaCl on survival of sublethally heat-injured Listeria monocytogenes.

PubMed

Williams, R C; Golden, D A

2001-03-20

The effect of package atmosphere on survival of uninjured and sublethally heat-injured Listeria monocytogenes, inoculated onto tryptose phosphate agar containing 0.85% lactic acid and 2% NaCl (TPALAS) was investigated. Inoculated TPALAS plates were packaged in air, 100% N2 (N2), 30% CO2-70% N2 (CO2-N2), and vacuum and stored at 4 and 20 degrees C for up to 31 days. Recovery of L. monocytogenes from TPALAS was influenced by the injury status (i.e., injured and uninjured) of the inoculum, storage atmosphere (air, N2, CO2-N2, and vacuum), storage temperature (4 and 20 degrees C), and recovery media [tryptose phosphate agar (TPA) and modified Oxford agar (MOX)] (P <0.05). Overall, storage at 4 degrees C supported greater survival than storage at 20 degrees C (P< 0.05). Uninjured L. monocytogenes stored at 4 degrees C was recovered on TPA better than sublethally heat-injured L. monocytogenes stored at 40 degrees C (P < 0.05). Recovery of sublethally heat-injured L. monocytogenes stored at 4 degrees C followed the order N2 > CO2-N2 > air > vacuum (P < 0.05), whereas recovery of uninjured L. monocyrogenes stored at 4 degrees C followed the order N2 > CO2-N2 > vacuum > air (P < 0.05). Air and vacuum atmospheres supported greater survival of uninjured and heat-injured L. monocytogenes than N2 and CO2-N2 atmospheres at 20 degrees C (P < 0.05). Recovery of sublethally heat-injured L. monocytogenes stored at 20 degrees C followed the order vacuum > air> CO2-N2 = N2 (P <0.05), whereas recovery of uninjured L. monocytogenes stored at 20 degrees C followed the order vacuum > air> CO2-N2 > N2 (P<0.05). Uninjured L. monocytogenes stored under N2 at 4 degrees C was recovered best, whereas sublethally heat-injured L. monocytogenes stored under N2 at 20 degrees C was recovered poorest (P < 0.05). Factors such as package atmosphere and storage temperature, involved in the production, storage, and distribution of fermented foods must be thoroughly evaluated when determining strategies for control and detection of L. monocytogenes in such products.

Auxiliary propulsion system flight package

NASA Technical Reports Server (NTRS)

Collett, C. R.

1987-01-01

Hughes Aircraft Company developed qualified and integrated flight, a flight test Ion Auxiliary Propulsion System (IAPS), on an Air Force technology satellite. The IAPS Flight Package consists of two identical Thruster Subsystems and a Diagnostic Subsystem. Each thruster subsystem (TSS) is comprised of an 8-cm ion Thruster-Gimbal-Beam Shield Unit (TGBSU); Power Electronics Unit; Digital Controller and Interface Unit (DCIU); and Propellant Tank, Valve and Feed Unit (PTVFU) plus the requisite cables. The Diagnostic Subsystem (DSS) includes four types of sensors for measuring the effect of the ion thrusters on the spacecraft and the surrounding plasma. Flight qualifications of IAPS, prior to installation on the spacecraft, consisted of performance, vibration and thermal-vacuum testing at the unit level, and thermal-vacuum testing at the subsystem level. Mutual compatibility between IAPS and the host spacecraft was demonstrated during a series of performance and environmental tests after the IAPS Flight Package was installed on the spacecraft. After a spacecraft acoustic test, performance of the ion thrusters was reverified by removing the TGBSUs for a thorough performance test at Hughes Research Laboratories (HRL). The TGBSUs were then reinstalled on the spacecraft. The IAPS Flight Package is ready for flight testing when Shuttle flights are resumed.

Advancements in meat packaging.

PubMed

McMillin, Kenneth W

2017-10-01

Packaging of meat provides the same or similar benefits for raw chilled and processed meats as other types of food packaging. Although air-permeable packaging is most prevalent for raw chilled red meat, vacuum and modified atmosphere packaging offer longer shelf life. The major advancements in meat packaging have been in the widely used plastic polymers while biobased materials and their integration into composite packaging are receiving much attention for functionality and sustainability. At this time, active and intelligent packaging are not widely used for antioxidant, antimicrobial, and other functions to stabilize and enhance meat properties although many options are being developed and investigated. The advances being made in nanotechnology will be incorporated into food packaging and presumably into meat packaging when appropriate and useful. Intelligent packaging using sensors for transmission of desired information and prompting of subsequent changes in packaging materials, environments or the products to maintain safety and quality are still in developmental stages. Copyright © 2017 Elsevier Ltd. All rights reserved.

Energy dependence corrections to MOSFET dosimetric sensitivity.

PubMed

Cheung, T; Butson, M J; Yu, P K N

2009-03-01

Metal Oxide Semiconductor Field Effect Transistors (MOSFET's) are dosimeters which are now frequently utilized in radiotherapy treatment applications. An improved MOSFET, clinical semiconductor dosimetry system (CSDS) which utilizes improved packaging for the MOSFET device has been studied for energy dependence of sensitivity to x-ray radiation measurement. Energy dependence from 50 kVp to 10 MV x-rays has been studied and found to vary by up to a factor of 3.2 with 75 kVp producing the highest sensitivity response. The detectors average life span in high sensitivity mode is energy related and ranges from approximately 100 Gy for 75 kVp x-rays to approximately 300 Gy at 6 MV x-ray energy. The MOSFET detector has also been studied for sensitivity variations with integrated dose history. It was found to become less sensitive to radiation with age and the magnitude of this effect is dependant on radiation energy with lower energies producing a larger sensitivity reduction with integrated dose. The reduction in sensitivity is however approximated reproducibly by a slightly non linear, second order polynomial function allowing corrections to be made to readings to account for this effect to provide more accurate dose assessments both in phantom and in-vivo.

Effect of packaging and storage temperature on the survival of Listeria monocytogenes inoculated postprocessing on sliced salami.

PubMed

Gounadaki, Antonia S; Skandamis, Panagiotis N; Drosinos, Eleftherios H; Nychas, George-John E

2007-10-01

The survival of postprocess Listeria monocytogenes contamination on sliced salami, stored under the temperatures associated with retail and domestic storage, was investigated. Sliced salami was inoculated with low and high concentrations of L. monocytogenes before being packaged under vacuum or air. Survival of L. monocytogenes was determined after storage of sausages for 45 or 90 days for low or high sample inocula, respectively, at 5, 15, and 25 degrees C. All survival curves of L. monocytogenes were characterized by an initial rapid inactivation within the first days of storage, followed by a second, slower inactivation phase or "tailing." Greater reduction of L. monocytogenes was observed at the high storage temperature (25 degrees C), followed by ambient (15 degrees C) and chill (5 degrees C) storage conditions. Moreover, vacuum packaging resulted in a slower destruction of L. monocytogenes than air packaging, and this effect increased as storage temperature decreased. Although L. monocytogenes numbers decreased to undetectable levels by the end of the storage period, the time (in days) needed for this reduction and for the total elimination of the pathogen decreased with high temperature, aerobic storage, and high inoculum. Results of this study clearly indicated that the kinetics of L. monocytogenes were highly dependent on the interaction of factors such as storage temperature, packaging conditions, and initial level of contamination (inoculum). These results may contribute to the exposure assessment of quantitative microbial risk assessment and to the establishment of storage-packaging recommendations of fermented sausages.

Very large scale heterogeneous integration (VLSHI) and wafer-level vacuum packaging for infrared bolometer focal plane arrays

NASA Astrophysics Data System (ADS)

Forsberg, Fredrik; Roxhed, Niclas; Fischer, Andreas C.; Samel, Björn; Ericsson, Per; Hoivik, Nils; Lapadatu, Adriana; Bring, Martin; Kittilsland, Gjermund; Stemme, Göran; Niklaus, Frank

2013-09-01

Imaging in the long wavelength infrared (LWIR) range from 8 to 14 μm is an extremely useful tool for non-contact measurement and imaging of temperature in many industrial, automotive and security applications. However, the cost of the infrared (IR) imaging components has to be significantly reduced to make IR imaging a viable technology for many cost-sensitive applications. This paper demonstrates new and improved fabrication and packaging technologies for next-generation IR imaging detectors based on uncooled IR bolometer focal plane arrays. The proposed technologies include very large scale heterogeneous integration for combining high-performance, SiGe quantum-well bolometers with electronic integrated read-out circuits and CMOS compatible wafer-level vacuum packing. The fabrication and characterization of bolometers with a pitch of 25 μm × 25 μm that are arranged on read-out-wafers in arrays with 320 × 240 pixels are presented. The bolometers contain a multi-layer quantum well SiGe thermistor with a temperature coefficient of resistance of -3.0%/K. The proposed CMOS compatible wafer-level vacuum packaging technology uses Cu-Sn solid-liquid interdiffusion (SLID) bonding. The presented technologies are suitable for implementation in cost-efficient fabless business models with the potential to bring about the cost reduction needed to enable low-cost IR imaging products for industrial, security and automotive applications.

Influence of heat processing on the volatile organic compounds and microbial diversity of salted and vacuum-packaged silver carp (Hypophthalmichthys molitrix) fillets during storage.

PubMed

Li, Dongping; Zhang, Jingbin; Song, Sijia; Feng, Ligeng; Luo, Yongkang

2018-06-01

Ready-to-eat products have become popular with most of the busy people in modern cities. Heat processing combined with vacuum-packaging is one of the most common methods to make ready-to-eat products with an extended shelf-life. In this study, the influence of heat processing [80 °C (LT) and 98 °C (HT) in water bath] on the quality of salted and vacuum-packaged silver carp (Hypophthalmichthys molitrix) fillets, stored at 20 ± 1 °C, was investigated by sensory analysis, biochemical analysis, and microbial diversity. SPME-GC/MS indicated the presence of 27 volatile organic compounds (VOCs) in fillets, and major VOCs were aldehydes and alcohols. Acids tended to increase during storage and caused a fetid odor at the end of storage. Culture-dependent method indicated that Bacillus dominated the spoiled LT and HT samples. In addition, Bacillus was identified as the main spoiler of deteriorated heated fillets by high-throughput sequencing. Sphingomonas and Brevibacillus dominated the indigenous bacteria of fresh raw fillets. After heat processing, LT samples exhibited higher organoleptic quality than HT samples on day 0. HT samples showed extended shelf-life at 20 °C storage compared to LT samples. Copyright © 2017. Published by Elsevier Ltd.

Effects of dietary magnesium and duration of refrigerated storage on the quality of vacuum-packaged, boneless pork loins.

PubMed

Apple, J K; Davis, J R; Rakes, L K; Maxwell, C V; Stivarius, M R; Pohlman, F W

2001-01-01

Quality data were initially collected on 78 pork loins from crossbred pigs fed diets containing 0, 1.25 or 2.5% magnesium mica (MM). Loins were then vacuum-packaged, and randomly assigned to either 4 or 8 weeks of storage at 2°C. Dietary MM had no (P > 0.05) effect on moisture loss/retention or subjective and objective color measurements. Purge volume increased (P<0.05) and drip loss decreased (P<0.05) as storage time increased. Moreover, longissimus thoracis et lumborum (LM) chops became lighter (P<0.05), redder (P<0.05), and more yellow (P<0.05) during 8 weeks of storage. Although TBARS values increased linearly (P<0.001) during extended storage, LM chops from pigs fed 2.5% MM tended to have lower (P<0.07) TBARS values after 4 weeks of storage than chops from pigs fed 0 and 1.25% MM. After 8 weeks of storage, however, there was a tendency for TBARS values of chops from pigs fed 1.25% MM to be lower (P<0.07) than chops from pigs fed 2.5% MM. Even though feeding swine diets containing MM did not affect color and water-holding capacity of pork loins during storage, the data indicated inclusion of MM in swine diets may retard onset of oxidative rancidity in vacuum-packaged pork loins.

NLC Electrical

Science.gov Websites

Racks and Cable Plant Instrumentation Systems Tunnel Electronics Enclosures Low Level RF Beam Positron Electronics Vacuum Electronics (Summary) System Notes NLC Electrical System Work Package Task Descriptions

A High-Q Resonant Pressure Microsensor with Through-Glass Electrical Interconnections Based on Wafer-Level MEMS Vacuum Packaging

PubMed Central

Luo, Zhenyu; Chen, Deyong; Wang, Junbo; Li, Yinan; Chen, Jian

2014-01-01

This paper presents a high-Q resonant pressure microsensor with through-glass electrical interconnections based on wafer-level MEMS vacuum packaging. An approach to maintaining high-vacuum conditions by integrating the MEMS fabrication process with getter material preparation is presented in this paper. In this device, the pressure under measurement causes a deflection of a pressure-sensitive silicon square diaphragm, which is further translated to stress build up in “H” type doubly-clamped micro resonant beams, leading to a resonance frequency shift. The device geometries were optimized using FEM simulation and a 4-inch SOI wafer was used for device fabrication, which required only three photolithographic steps. In the device fabrication, a non-evaporable metal thin film as the getter material was sputtered on a Pyrex 7740 glass wafer, which was then anodically bonded to the patterned SOI wafer for vacuum packaging. Through-glass via holes predefined in the glass wafer functioned as the electrical interconnections between the patterned SOI wafer and the surrounding electrical components. Experimental results recorded that the Q-factor of the resonant beam was beyond 22,000, with a differential sensitivity of 89.86 Hz/kPa, a device resolution of 10 Pa and a nonlinearity of 0.02% F.S with the pressure varying from 50 kPa to 100 kPa. In addition, the temperature drift coefficient was less than −0.01% F.S/°C in the range of −40 °C to 70 °C, the long-term stability error was quantified as 0.01% F.S over a 5-month period and the accuracy of the microsensor was better than 0.01% F.S. PMID:25521385

A high-Q resonant pressure microsensor with through-glass electrical interconnections based on wafer-level MEMS vacuum packaging.

PubMed

Luo, Zhenyu; Chen, Deyong; Wang, Junbo; Li, Yinan; Chen, Jian

2014-12-16

This paper presents a high-Q resonant pressure microsensor with through-glass electrical interconnections based on wafer-level MEMS vacuum packaging. An approach to maintaining high-vacuum conditions by integrating the MEMS fabrication process with getter material preparation is presented in this paper. In this device, the pressure under measurement causes a deflection of a pressure-sensitive silicon square diaphragm, which is further translated to stress build up in "H" type doubly-clamped micro resonant beams, leading to a resonance frequency shift. The device geometries were optimized using FEM simulation and a 4-inch SOI wafer was used for device fabrication, which required only three photolithographic steps. In the device fabrication, a non-evaporable metal thin film as the getter material was sputtered on a Pyrex 7740 glass wafer, which was then anodically bonded to the patterned SOI wafer for vacuum packaging. Through-glass via holes predefined in the glass wafer functioned as the electrical interconnections between the patterned SOI wafer and the surrounding electrical components. Experimental results recorded that the Q-factor of the resonant beam was beyond 22,000, with a differential sensitivity of 89.86 Hz/kPa, a device resolution of 10 Pa and a nonlinearity of 0.02% F.S with the pressure varying from 50 kPa to 100 kPa. In addition, the temperature drift coefficient was less than -0.01% F.S/°C in the range of -40 °C to 70 °C, the long-term stability error was quantified as 0.01% F.S over a 5-month period and the accuracy of the microsensor was better than 0.01% F.S.

Effect of age of cook-in-bag delicatessen meats formulated with lactate-diacetate on the behavior of Listeria monocytogenes contamination introduced when opening the packages during storage.

PubMed

Geornaras, Ifigenia; Toczko, Darren; Sofos, John N

2013-07-01

This study evaluated the potential effect of age of cook-in-bag ham and turkey breast delicatessen meats formulated with lactate-diacetate on survival and/or growth of Listeria monocytogenes introduced after opening of packages and slicing of product. Commercially prepared cured ham and turkey breast products formulated with potassium lactate and sodium diacetate were stored at 1.7°C unsliced, in their original cook-in-bags, and without postlethality exposure. On days 5, 90, 120, and 180 of storage, product slices (10.2 by 7.6 cm) were surface inoculated (1 to 2 log CFU/cm²) with a 10-strain mixture of L. monocytogenes, vacuum packaged (seven slices per bag), and stored at 4°C for up to 13 weeks. Inoculated levels of L. monocytogenes on both products were 1.4 to 1.5 log CFU/cm². Irrespective of product age at slicing and inoculation, after 13 weeks of vacuum-packaged storage (4°C), pathogen counts on product slices were 1.5 to 2.3 (ham) and 2.3 to 2.5 (turkey) log CFU/cm². Overall, the results of the study showed that the age of the cook-in-bag products prior to slicing and inoculation with the pathogen did not (P ≥ 0.05) affect the behavior of L. monocytogenes during vacuum-packaged storage (4°C, up to 13 weeks) of ham and turkey slices. Mean counts of lactic acid bacteria and yeasts and molds, when detected, did not exceed approximately 1 and 2 log CFU/cm², respectively, among all stored samples. Findings of the study will be useful to the meat industry and risk assessors in their efforts to control L. monocytogenes in ready-to-eat meat products.

Use of Optical Oxygen Sensors in Non-Destructively Determining the Levels of Oxygen Present in Combined Vacuum and Modified Atmosphere Packaged Pre-Cooked Convenience-Style Foods and the Use of Ethanol Emitters to Extend Product Shelf-Life

PubMed Central

Hempel, Andreas W.; Papkovsky, Dmitri B.; Kerry, Joseph P.

2013-01-01

O2 sensors were used to non-destructively monitor O2 levels in commercially packed pre-cooked, convenience modified atmosphere packaging (MAP) foods. A substantial level of O2 (>15%) was present in packs resulting in a shorter than expected shelf-life, where the primary spoilage mechanism was found to be mould. Various combinations of vacuum (0–0.6 MPa) and gas flush (0.02–0.03 MPa) (30% CO2/70% N2) settings were assessed as treatments that result in the desired shelf-life (28 days). This was achieved using the combined treatment of vacuum 0.35 MPa and gas flush 0.02 MPa which resulted in a reduction of 6%–9% O2 in all three samples (battered sausages (BS), bacon slices (BA), and meat and potato pies (PP)). Reduced O2 levels reflect the microbial quality of products, which has been successfully reduced. Duplicate samples of all product packs were produced using ethanol emitters (EE) to see if shelf-life could be further extended. Results showed a further improvement in shelf-life to 35 days. Sensory analysis showed that ethanol flavour and aroma was not perceived by panellists in two of the three products assessed. This study demonstrates how smart packaging technologies, both intelligent and active, can be used to assist in the modification of conventional packaging systems in order to enhance product quality and safety and through the extension of product shelf-life. PMID:28239134

Shelf-life extension of vacuum-packaged meat from pheasant (Phasianus colchicus) by lactic acid treatment.

PubMed

Pfeifer, Agathe; Smulders, Frans J M; Paulsen, Peter

2014-07-01

We investigated the influence of lactic acid treatment of pheasant meat before vacuum-packaged storage of 3, 7, and 10 d at +6°C on microbiota and pH. Breast muscle samples were collected from carcasses of slaughtered as well as from hunted (shot) wild pheasants. Immersion of meat samples in 3% (wt/wt) lactic acid for 60 s effectuated a significant drop in pH of approximately 0.5 to 0.7 units, which remained during the entire storage period. In parallel, total aerobic counts of such treated and stored samples were on an average 1.5 to 1.7 log units lower than in non-acid-treated samples. Similar results were found for Enterobacteriaceae. A significant decrease in pH was measured at d 7 and 10 in the acid-treated samples in comparison with the untreated ones. In summary, the immersion of pheasant breast meat cuts in dilute lactic acid significantly reduced microbiota during vacuum-packed storage, even at slight temperature abuse conditions. © 2014 Poultry Science Association Inc.

Influence of PA6 nanocomposite films on the stability of vacuum-aged beef loins during storage in modified atmospheres.

PubMed

Picouet, P A; Fernandez, A; Realini, C E; Lloret, E

2014-01-01

A masterbatch of polyamide 6 (PA6) containing dispersed nanoclays, was used to fabricate a novel multilayer film for vacuum packed meat. Performance of the nanocomposite was compared to a control PA6 multilayer and a high barrier commercial film. Addition of nanoclays improved oxygen barrier properties, UV-blocking capability and stiffness. Beef loins were vacuum-aged using the three films for 0 7, 14 and 21 days at 2°C. After each ageing time, beef steaks were packaged in commercial trays and high oxygen atmosphere and stored at 4°C for 9 days. Beef quality parameters and gas content were studied during display time in MAP (1, 3, 6 and 9 d). Beef quality parameters were not influenced by the packaging materials used during ageing and the performance of nanocomposites was comparable to high barrier films. Ageing had a positive impact on the stabilization of redness up to day 6 in MAP. Thereafter, oxymyoglobin content and oxidation levels were negatively influenced by ageing. © 2013.

Focusing on Environmental Biofilms With Variable-Pressure Scanning Electron Microscopy

NASA Astrophysics Data System (ADS)

Joubert, L.; Wolfaardt, G. M.; Du Plessis, K.

2006-12-01

Since the term biofilm has been coined almost 30 years ago, visualization has formed an integral part of investigations on microbial attachment. Electron microscopic (EM) biofilm studies, however, have been limited by the hydrated extracellular matrix which loses structural integrity with conventional preparative techniques, and under required high-vacuum conditions, resulting in a loss of information on spatial relationships and distribution of biofilm microbes. Recent advances in EM technology enable the application of Variable Pressure Scanning Electron Microscopy (VP SEM) to biofilms, allowing low vacuum and hydrated chamber atmosphere during visualization. Environmental biofilm samples can be viewed in situ, unfixed and fully hydrated, with application of gold-sputter-coating only, to increase image resolution. As the impact of microbial biofilms can be both hazardous and beneficial to man and his environment, recognition of biofilms as a natural form of microbial existence is needed to fully assess the potential role of microbial communities on technology. The integration of multiple techniques to elucidate biofilm processes has become imperative for unraveling complex phenotypic adaptations of this microbial lifestyle. We applied VP SEM as integrative technique with traditional and novel analytical techniques to (1)localize lignocellulosic microbial consortia applied for producing alternative bio-energy sources in the mining wastewater industry, (2) characterize and visualize wetland microbial communities in the treatment of winery wastewater, and (3)determine the impact of recombinant technology on yeast biofilm behavior. Visualization of microbial attachment to a lignocellulose substrate, and degradation of exposed plant tissue, gave insight into fiber degradation and volatile fatty acid production for biological sulphate removal from mining wastewater. Also, the 3D-architecture of complex biofilms developing in constructed wetlands was correlated with molecular fingerprints of wetland communities using tRFLP (Terminal Restriction Fragment Length Polymorphism) - and gave evidence of temporal and spatial variation in a wetland system, to potentially be applied as management tool in wastewater treatment. Visualization of differences in biofilm development by wild and recombinant yeast strains furthermore supported real-time quantitative data of biofilm development by Cryptococcus laurentii and Saccharomyces yeast strains. In all cases VP SEM allowed a more holistic interpretation of biofilm processes than afforded by quantitative empirical data only.

The influence of headspace and dissolved oxygen level on growth and haemolytic BL enterotoxin production of a psychrotolerant Bacillus weihenstephanensis isolate on potato based ready-to-eat food products.

PubMed

Samapundo, S; Everaert, H; Wandutu, J N; Rajkovic, A; Uyttendaele, M; Devlieghere, F

2011-04-01

The major objective of this study was to determine the influence of the initial headspace and dissolved O(2) level and vacuum packaging on growth and diarrhoeal enterotoxin production by Bacillus weihenstephanensis on potato based ready-to-eat food products. In general, the lower the initial headspace or dissolved O(2) level the slower the maximum growth rate (μ(max), log(10) CFU g(-1) d(-1)), the longer the lag phase duration (λ, d) and the smaller the maximum population density (N(max), log(10) CFU g(-1)) became. The slowest μ(max), the longest λ and the smallest N(max) were generally found for growth under vacuum packaging. This implies shorter shelf-lives will occur at higher initial headspace or dissolved O(2) levels as the growth of B. weihenstephanensis to the infective dose of 10(5) CFU g(-1) in such atmospheres takes a shorter time. Significant consumption of dissolved O(2) only occurred when growth shifted from the lag to the exponential phase and growth generally transitioned from the exponential to the stationary phase when the dissolved O(2) levels fell below ca. 75 ppb. Diarrhoeal enterotoxin production (determined via detection of the L2 component of haemolytic BL) was similar for growth under initial headspace O(2) levels of 1-20.9%, and was only reduced when growth took place under vacuum packaging. The reduction in L2 production when growth took place under vacuum was most probably related to the low final cell densities observed under this condition. Both growth and L2 production were inhibited over a 32-day incubation period at 7 °C by 40% CO(2) irrespective of the headspace or dissolved O(2) levels. The results illustrate the importance of residual O(2) and CO(2) on the shelf-stability and safety of modified atmosphere packaged potato based ready-to-eat food products with regards to B. weihenstephanensis. Copyright © 2010 Elsevier Ltd. All rights reserved.

Effect of Hops Beta Acids on the Survival of Unstressed- or Acid-Stress-Adapted-Listeria Monocytogenes and on the Quality and Sensory Attributes of Commercially Cured Ham Slices.

PubMed

Wang, Li; McKeith, Amanda Gipe; Shen, Cangliang; Carter, Kelsey; Huff, Alyssa; McKeith, Russell; Zhang, Xinxia; Chen, Zhengxing

2016-02-01

This study evaluated the antilisterial activity of hops beta acids (HBA) and their impact on the quality and sensory attributes of ham. Commercially cured ham slices were inoculated with unstressed- and acid-stress-adapted (ASA)-L. monocytogenes (2.2 to 2.5 log CFU/cm(2) ), followed by no dipping (control), dipping in deionized (DI) water, or dipping in a 0.11% HBA solution. This was followed by vacuum or aerobic packaging and storage (7.2 °C, 35 or 20 d). Samples were taken periodically during storage to check for pH changes and analyze the microbial populations. Color measurements were obtained by dipping noninoculated ham slices in a 0.11% HBA solution, followed by vacuum packaging and storage (4.0 °C, 42 d). Sensory evaluations were performed on ham slices treated with 0.05% to 0.23% HBA solutions, followed by vacuum packaging and storage (4.0 °C, 30 d). HBA caused immediate reductions of 1.2 to 1.5 log CFU/cm(2) (P < 0.05) in unstressed- and ASA-L. monocytogenes populations on ham slices. During storage, the unstressed-L. monocytogenes populations on HBA-treated samples were 0.5 to 2.0 log CFU/cm(2) lower (P < 0.05) than control samples and those dipped in DI water. The lag-phase of the unstressed-L. monocytogenes population was extended from 3.396 to 7.125 d (control) to 7.194 to 10.920 d in the HBA-treated samples. However, the ASA-L. monocytogenes population showed resistance to HBA because they had a higher growth rate than control samples and had similar growth variables to DI water-treated samples during storage. Dipping in HBA solution did not adversely affect the color or sensory attributes of the ham slices stored in vacuum packages. These results are useful for helping ready-to-eat meat processors develop operational procedures for applying HBA on ham slices. © 2016 Institute of Food Technologists®

Inhibitory effects of cinnamon and clove essential oils on mold growth on baked foods.

PubMed

Ju, Jian; Xu, Xiaomiao; Xie, Yunfei; Guo, Yahui; Cheng, Yuliang; Qian, He; Yao, Weirong

2018-02-01

This study evaluated the minimum inhibition concentration (MIC) and minimum lethal concentration (MLC) of cinnamon and clove essential oils against mold growth on green bean cake and finger citron crisp cake, and also examined the effects of these two essential oils and their application methods on the shelf life of the baked products in normal and vacuum packages by accelerated storage test. The results showed that the MIC of cinnamon and clove essential oils against molds were 0.21-0.83 and 0.21-1.67μL/mL, respectively and the MLC were 0.42-0.83 and 0.83-1.67μL/mL, respectively. In normal package cinnamon and clove essential oils could prolong the shelf life of green bean cake 9-10 and 3-4days, respectively and could prolong the shelf life of finger citron crisp cake 5-6 and 2-3days, respectively. And in vacuum package they were 15-16, 8-9, 10-12 and 7-9days, respectively in turn. Copyright © 2017. Published by Elsevier Ltd.

Evaluation of the effectiveness of non-irradiated and chlorine-free packaging for fresh beef preservation.

PubMed

Rodrigues, José B M; Sarantópoulos, Claire I G L; Bromberg, Renata; Andrade, Juliana C; Brunelli, Kleber; Miyagusku, Luciana; Marquezini, Miriam G; Yamada, Eunice A

2017-03-01

This study evaluates the potential of using non-irradiated barrier-shrink bags containing ethylene-vinyl alcohol copolymer (EVOH), polyamide (PA) and ethylene ionomer in their structures to preserve vacuum-packaged fresh beef as an alternative to traditional gamma-ray cross-linked bags containing polyvinylidene chloride (PVDC). Boneless beef rib eye roll cuts were vacuum-packed in an industrial processing plant using EVOH 44% mol, EVOH 32% mol and a control PVDC barrier shrink bags. The cuts were evaluated during storage at 0.5°C. The EVOH films presented similar performance compared to control PVDC barrier shrink bags related to bacteria growth and purge loss. Packages with EVOH 32% mol film presented better performance than control bag with respect to the meat sensorial attributes, including fewer bubbles and better adhesion. EVOH 44% mol bags presented the highest rate of colour loss. The EVOH 32% mol non-irradiated and chlorine-free film is as effective for the preservation of fresh beef as traditional PVDC-irradiated shrink bags. Copyright © 2016 Elsevier Ltd. All rights reserved.

Improvements in the shelf life of commercial corn dry masa flour (CMF) by reducing lipid oxidation.

PubMed

Márquez-Castillo, A; Vidal-Quintanar, R L

2011-03-01

To improve the shelf life of commercial nixtamalized corn dry masa flour (CMF), the modified atmosphere packaging (MAP) was used. Pouches (20 × 20 cm) of ethyl vinyl alcohol (EVOH) with 180 g of CMF were stored at 55 °C, and a(w) of 0.45; under Light and Dark conditions, antioxidants (0.02% TBHQ), Vacuum, and N(2) and CO(2), and used as treatments. Thereafter, changes in their linoleic acid (LA) concentration by GC, peroxide (PV), and anisidine values (p-A), which were monitored for 180 d. EVOH showed a significantly lower consumption of LA by autoxidation (11.7% ± 0.2% in 117 d) than polyethylene film (70.5% ± 0.3% in 113 d) under the same storage temperature. The elimination of oxygen by vacuum in each pouch allowed a low consumption (16.4% ± 0.1%) of LA. PV (14.5 ± 0.09 mEq/kg of fat), and p-A (63 ± 0.16 mmol/kg) were low, and generated over 121 d of storage. CMF stored under MAP had 100% protection against oxidation of LA. A combination of Vacuum and EVOH packaging extended the shelf life of CMF to 108 d with only 10% of LA loss. For retail stores, the EVOH packaging will reduce lipid oxidation of CMF and safety related to off odors and flavors from the oxidation of tortillas will increase dramatically.

Occurrence and role of lactic acid bacteria in seafood products.

PubMed

Françoise, Leroi

2010-09-01

Lactic acid bacteria (LAB) in fish flesh has long been disregarded because the high post-mortem pH, the low percentage of sugars, the high content of low molecular weight nitrogenous molecules and the low temperature of temperate waters favor the rapid growth of pH-sensitive psychrotolerant marine Gram-negative bacteria like Pseudomonas, Shewanella and Photobacterium. In seafood packed in both vacuum (VP) and modified atmosphere (MAP) packaging commonly CO(2) enriched, the growth of the Gram-negative aerobic bacteria group (predominantly pseudomonads) is effectively inhibited and the number reached by LAB during storage is higher than that achieved in air but always several log units lower than the trimethylamine oxide (TMA-O) reducing and CO(2)-resistant organisms (Shewanella putrefaciens and Photobacterium phosphoreum). Accordingly, LAB are not of much concern in seafood neither aerobically stored nor VP and MAP. However, they may acquire great relevance in lightly preserved fish products (LPFP), including those VP or MAP. Fresh fish presents a very high water activity (aw) value (0.99). However, aw is reduced to about 0.96 when salt (typically 6% WP) is added to the product. As a result, aerobic Gram-negative bacteria are inhibited, which allows the growth of other organisms more resistant to reduced aw, i.e. LAB, and then they may acquire a central role in the microbial events occurring in the product. Changes in consumers' habits have led to an increase of convenient LPFP with a relative long shelf-life (at least 3 weeks) which, on the other hand, may constitute a serious problem from a safety perspective since Listeria monocytogenes and sometimes Clostridium botulinum (mainly type E) may able to grow. In any case the LAB function in marine products is complex, depending on species, strains, interaction with other bacteria and the food matrix. They may have no particular effect or they may be responsible for spoilage and, in certain cases, they may even exert a bioprotective effect in relation to undesirable bacteria. The bioprotective potential of endogenous LAB in relation to pathogens and spoiling bacteria has often been highlighted. However, the technology is still in its infancy compared with foods dairy and meat products in which either the carbohydrate content (dairy products) or sugar and salt added (meat products) favor the acidification by LAB that enable a natural preservation of the product. Successful studies on LAB as probiotic for fish intensify, but this potential is still to be explored for human. Although not usual, some applications of LAB for fermentation of marine products and by-products are described. Copyright 2010 Elsevier Ltd. All rights reserved.

Preservation of fresh meat with active and modified atmosphere packaging conditions.

PubMed

Skandamis, Panagiotis N; Nychas, George-John E

2002-11-15

The sensory, microbiological and physicochemical attributes of fresh meat stored at 5 and 15 degrees C were affected by the combined effect of volatile compounds of oregano essential oil and modified atmosphere packaging conditions (40% CO2/30% N2/30% O2, 100% CO2, 80% CO2/20% air, vacuum pack and air). It was found that the extension of shelf life of meat samples depended on the packaging conditions and augmented in the order: air < vacuum pack < 40% CO2/30% N2/30% O2 < 80% CO2/ 20% air < 100% CO2. Longer shelf life was observed in samples supplemented with the volatile compounds of oregano essential oil and stored under the same packaging conditions mentioned above. The extension of shelf life may be due to the synergistic effect of volatile compounds of oregano essential oil and the modified atmosphere packaging used on the microbiological and physicochemical characteristics of meat. Indeed, both these hurdles can prolong and delay microbial growth or suppress the final counts of the spoilage microorganisms in comparison with the 'control' samples. The effect of essential oil volatile compounds was even more pronounced on the physicochemical changes of meat samples caused by microbial association. Oregano essential oil delayed glucose and lactate consumption, both indicators of meat spoilage aerobically as well as under 40% CO2/30% N2/30% O2, and 100% CO2. Finally, changes in other metabolites such as formic acid were also observed.

Effect of Addition of Natural Antioxidants on the Shelf-Life of “Chorizo”, a Spanish Dry-Cured Sausage

PubMed Central

Pateiro, Mirian; Bermúdez, Roberto; Lorenzo, José Manuel; Franco, Daniel

2015-01-01

The dose effect of the addition of natural antioxidants (tea, chestnut, grape seed and beer extracts) on physicochemical, microbiological changes and on oxidative stability of dry-cured “chorizo”, as well as their effect during the storage under vacuum conditions was evaluated. Color parameters were significantly (p < 0.05) affected by the addition of antioxidants so that samples that contained antioxidants were more effective in maintaining color. The improving effects were dose-dependent with highest values with the dose of 50 mg/kg during ripening and depend on the extract during vacuum packaging. Addition of antioxidants decreased (p < 0.05) the oxidation, showing thiobarbituric acid reactive substances (TBARS) values below 0.4 mg MDA/kg. Natural antioxidants matched or even improved the results obtained for butylated hydroxytoluene (BHT). Regarding texture profile analysis (TPA) analysis, hardness values significantly (p < 0.001) decreased with the addition of antioxidants, obtaining the lower results with the dose of 200 mg/kg both during ripening and vacuum packaging. Antioxidants reduced the counts of total viable counts (TVC), lactic acid bacteria (LAB), mold and yeast. Free fatty acid content during ripening and under vacuum conditions showed a gradual and significant (p < 0.05) release as a result of lipolysis. At the end of ripening, the addition of GRA1000 protected chorizos from oxidative degradation. PMID:26785337

Encapsulation for smart textile electronics - humidity and temperature sensor.

PubMed

Larsson, Andreas; Tran, Thanh-Nam; Aasmundtveit, Knut E; Seeberg, Trine M

2015-01-01

A combined humidity and temperature sensor was packaged by vacuum casting onto three different types of textiles; cotton, nylon and a waterproof fabric. This was done in order to integrate the sensor in a jacket in a soft and reliable way without changing the sensor performance. A membrane was custom made and integrated into the device to protect the sensor from the environment. The packaged sensors performance was characterized in a climate chamber were the relative humidity and temperature ranged from 25 % to 95 % and -10 °C to 75 °C respectively. The packaged sensors showed insignificant to limited performance degradation.

European consumer response to packaging technologies for improved beef safety.

PubMed

Van Wezemael, Lynn; Ueland, Øydis; Verbeke, Wim

2011-09-01

Beef packaging can influence consumer perceptions of beef. Although consumer perceptions and acceptance are considered to be among the most limiting factors in the application of new technologies, there is a lack of knowledge about the acceptability to consumers of beef packaging systems aimed at improved safety. This paper explores European consumers' acceptance levels of different beef packaging technologies. An online consumer survey was conducted in five European countries (n=2520). Acceptance levels among the sample ranged between 23% for packaging releasing preservative additives up to 73% for vacuum packaging. Factor analysis revealed that familiar packaging technologies were clearly preferred over non-familiar technologies. Four consumer segments were identified: the negative (31% of the sample), cautious (30%), conservative (17%) and enthusiast (22%) consumers, which were profiled based on their attitudes and beef consumption behaviour. Differences between consumer acceptance levels should be taken into account while optimising beef packaging and communicating its benefits. Copyright © 2011 Elsevier Ltd. All rights reserved.

16 CFR 501.3 - Replacement bags for vacuum cleaners.

Code of Federal Regulations, 2010 CFR

2010-01-01

... statement appears on the principal display panel of the package accurately identifying the make and model of...) The name and place of business of the manufacturer, packer, or distributor of the replacement bags, in...

Effect of packaging materials on the chemical composition and microbiological quality of edible mushroom (Pleurotus ostreatus) grown on cassava peels.

PubMed

Ajayi, Oluwakemi; Obadina, Adewale; Idowu, Micheal; Adegunwa, Mojisola; Kajihausa, Olatundun; Sanni, Lateef; Asagbra, Yemisi; Ashiru, Bolanle; Tomlins, Keith

2015-07-01

Edible fungi such as mushrooms are highly perishable and deteriorate few days after harvest due to its high moisture content and inability to maintain their physiological status. In this study, the effect of packaging materials on the nutritional composition of mushroom cultivated from cassava peels was investigated. Mushroom samples were dried at 50°C in a cabinet dryer for 8 h. The dried mushroom samples packaged in four different packaging materials; high density polyethylene (HDPE), polypropylene (PP), laminated aluminum foil (LAF), high density polyethylene under vacuum (HDPEV) were stored at freezing (0°C) temperatures for 12 weeks. Samples were collected at 2-week intervals and analyzed for proximate composition (carbohydrate, protein, fat, fiber, ash, moisture), mineral content (calcium, potassium), vitamin C content, and microbiological qualities (total aerobic count, Pseudomonal count, Coliform count, Staphylococcal count, Salmonella count) using the standard laboratory procedures. Carbohydrate, protein, fat content of dried mushrooms packaged in HDPE at freezing temperature ranged from 45.2% to 53.5%, 18.0% to 20.3%, and 3.2% to 4.3%, while mushrooms in polypropylene ranged from 45.2% to 53.5%, 18.5% to 20.3%, 2.6% to 4.3%. Carbohydrate, protein, fat of mushroom in LAF ranged from 47.8% to 53.5%, 17.3% to 20.3%, and 3.3% to 4.3%, respectively, while carbohydrate, protein, fat of mushroom in HDPEV ranged from 51.1% to 53.5%, 19.5% to 20.3%, and 3.5% to 4.3%. Microbiological analysis showed that total aerobic count, Pseudomonal count, and Staphyloccocal count of dried mushroom ranged from 2.3 to 3.8 log cfu/g, 0.6 to 1.1 log cfu/g, and 0.4 to 0.5 log cfu/g, respectively. In conclusion, dried mushroom in HDPE packaged under vacuum at freezing temperature retained the nutritional constituents than those packaged with other packaging materials.

Effect of packaging materials on the chemical composition and microbiological quality of edible mushroom (Pleurotus ostreatus) grown on cassava peels

PubMed Central

Ajayi, Oluwakemi; Obadina, Adewale; Idowu, Micheal; Adegunwa, Mojisola; Kajihausa, Olatundun; Sanni, Lateef; Asagbra, Yemisi; Ashiru, Bolanle; Tomlins, Keith

2015-01-01

Edible fungi such as mushrooms are highly perishable and deteriorate few days after harvest due to its high moisture content and inability to maintain their physiological status. In this study, the effect of packaging materials on the nutritional composition of mushroom cultivated from cassava peels was investigated. Mushroom samples were dried at 50°C in a cabinet dryer for 8 h. The dried mushroom samples packaged in four different packaging materials; high density polyethylene (HDPE), polypropylene (PP), laminated aluminum foil (LAF), high density polyethylene under vacuum (HDPEV) were stored at freezing (0°C) temperatures for 12 weeks. Samples were collected at 2-week intervals and analyzed for proximate composition (carbohydrate, protein, fat, fiber, ash, moisture), mineral content (calcium, potassium), vitamin C content, and microbiological qualities (total aerobic count, Pseudomonal count, Coliform count, Staphylococcal count, Salmonella count) using the standard laboratory procedures. Carbohydrate, protein, fat content of dried mushrooms packaged in HDPE at freezing temperature ranged from 45.2% to 53.5%, 18.0% to 20.3%, and 3.2% to 4.3%, while mushrooms in polypropylene ranged from 45.2% to 53.5%, 18.5% to 20.3%, 2.6% to 4.3%. Carbohydrate, protein, fat of mushroom in LAF ranged from 47.8% to 53.5%, 17.3% to 20.3%, and 3.3% to 4.3%, respectively, while carbohydrate, protein, fat of mushroom in HDPEV ranged from 51.1% to 53.5%, 19.5% to 20.3%, and 3.5% to 4.3%. Microbiological analysis showed that total aerobic count, Pseudomonal count, and Staphyloccocal count of dried mushroom ranged from 2.3 to 3.8 log cfu/g, 0.6 to 1.1 log cfu/g, and 0.4 to 0.5 log cfu/g, respectively. In conclusion, dried mushroom in HDPE packaged under vacuum at freezing temperature retained the nutritional constituents than those packaged with other packaging materials. PMID:26288720

New Finsler package

NASA Astrophysics Data System (ADS)

Youssef, Nabil L.; Elgendi, S. G.

2014-03-01

The book “Handbook of Finsler geometry” has been included with a CD containing an elegant Maple package, FINSLER, for calculations in Finsler geometry. Using this package, an example concerning a Finsler generalization of Einstein’s vacuum field equations was treated. In this example, the calculation of the components of the hv-curvature of Cartan connection leads to wrong expressions. On the other hand, the FINSLER package works only in dimension four. We introduce a new Finsler package in which we fix the two problems and solve them. Moreover, we extend this package to compute not only the geometric objects associated with Cartan connection but also those associated with Berwald, Chern and Hashiguchi connections in any dimension. These improvements have been illustrated by a concrete example. Furthermore, the problem of simplifying tensor expressions is treated. This paper is intended to make calculations in Finsler geometry more easier and simpler.

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

Martin, R; Pan, T; Li, B

Purpose: Dual energy CT has a variety of uses in a small animal setting including quantification and enhanced visualization of contrast agent. This study aims to determine the best energy combinations for contrast enhanced DECT on the XRAD 225Cx (Precision x-ray), a small animal IGRT system with a nominal energy range of 20 – 225 kVp. Focus was placed on material density accuracy and low contrast detectability. Methods: Simulations of single energy scans of an object containing concentrations of iodine varying from 0.5 to 50 mg/ml were performed using the simulation package ImaSim. Energy spectra from 50 – 220 kVpmore » were calculated using the same software. For approximate Poisson noise modeling, mAs were chosen such that 30% of the total 10cGy dose was assigned to the low energy scan. A calibration involving projections of objects containing different thicknesses of iodine (0–0.5 mm) and water (0–50 mm) was performed for each energy and fit to a cubic equation as the calibration curve for each energy pair. Results: Contrast to noise ratios of the iodine material images and accuracies in iodine density measurements were measured. Gradual improvements in each metric were seen with increasing high energy. Larger improvements in CNR were observed for decreasing the low energy. Errors in iodine density were generally close to 5% for concentrations of iodine above 3 mg/ml but increased to around 15% for 50 kVp, likely due to its proximity to the discontinuity caused by the k-edge of iodine. Conclusion: Based on these simulations, the best energy combination for detecting low concentrations of iodine using a projection space calibration procedure is 50/200 kVp. However, if accuracy is most important 80/220 kVp is ideal, with 60/220 kVp being a good compromise to achieve both goals. Future work is necessary to verify these conclusions with physical data.« less

Microbial deterioration of vacuum-packaged chilled beef cuts and techniques for microbiota detection and characterization: a review.

PubMed

Hernández-Macedo, Maria Lucila; Barancelli, Giovana Verginia; Contreras-Castillo, Carmen Josefina

2011-01-01

Gas production from microbial deterioration in vacuum-packs of chilled meat leads to pack distension, which is commonly referred as blown pack. This phenomenon is attributed to some psychrophilic and psychrotrophic Clostridium species, as well as Enterobacteria. The ability of these microorganisms to grow at refrigeration temperatures makes the control by the meat industry a challenge. This type of deterioration has been reported in many countries including some plants in the Midwestern and Southeastern regions of Brazil. In addition to causing economic losses, spoilage negatively impacts the commercial product brand, thereby impairing the meat industry. In the case of strict anaerobes species they are difficult to grow and isolate using culture methods in conventional microbiology laboratories. Furthermore, conventional culture methods are sometimes not capable of distinguishing species or genera. DNA-based molecular methods are alternative strategies for detecting viable and non-cultivable microorganisms and strict anaerobic microorganisms that are difficult to cultivate. Here, we review the microorganisms and mechanisms involved in the deterioration of vacuum-packaged chilled meat and address the use of molecular methods for detecting specific strict anaerobic microorganisms and microbial communities in meat samples.

Heat transfer implications in the first MEMS fabricated thermal transiration-driven vacuum pump for gases

NASA Technical Reports Server (NTRS)

Vargo, S. E.; Green, A. A.; Muntz, E. P.

2000-01-01

The success of NASA's future space missions and the development of portable, commercial instrument packages will depend greatly on miniaturized components enabled by the use of microelectromechanical systems (MEMS).

Impact of high pressure treatment and intramuscular fat content on colour changes and protein and lipid oxidation in sliced and vacuum-packaged Iberian dry-cured ham.

PubMed

Fuentes, Verónica; Utrera, Mariana; Estévez, Mario; Ventanas, Jesús; Ventanas, Sonia

2014-08-01

The effect of high hydrostatic pressure (HHP) (600MPa) and intramuscular fat content (IMF) on colour parameters and oxidative stability of lipids and proteins in sliced vacuum-packaged Iberian dry-cured ham during refrigerated storage (120 days at 2°C) was investigated. Several studies have investigated the influence of HHP on lipid oxidation of meat products. However, its effects on protein carbonylation, as also the influence of IMF content on this carbonylation are poorly understood. HHP treatment had a significant effect on lean lightness after 0 and 120 days of storage while IMF content increased lightness and yellowness over time. Regarding oxidative stability, the effect of HHP treatment depended on IMF content samples with a high IMF having greater lipid instability while samples with a low IMF underwent more protein carbonylation. Copyright © 2014 Elsevier Ltd. All rights reserved.

Space ultra-vacuum facility and method of operation

NASA Technical Reports Server (NTRS)

Naumann, Robert J. (Inventor)

1988-01-01

A wake shield space processing facility (10) for maintaining ultra-high levels of vacuum is described. The wake shield (12) is a truncated hemispherical section having a convex side (14) and a concave side (24). Material samples (68) to be processed are located on the convex side of the shield, which faces in the wake direction in operation in orbit. Necessary processing fixtures (20) and (22) are also located on the convex side. Support equipment including power supplies (40, 42), CMG package (46) and electronic control package (44) are located on the convex side (24) of the shield facing the ram direction. Prior to operation in orbit the wake shield is oriented in reverse with the convex side facing the ram direction to provide cleaning by exposure to ambient atomic oxygen. The shield is then baked-out by being pointed directed at the sun to obtain heating for a suitable period.

Vacuum mechatronics

NASA Technical Reports Server (NTRS)

Hackwood, Susan; Belinski, Steven E.; Beni, Gerardo

1989-01-01

The discipline of vacuum mechatronics is defined as the design and development of vacuum-compatible computer-controlled mechanisms for manipulating, sensing and testing in a vacuum environment. The importance of vacuum mechatronics is growing with an increased application of vacuum in space studies and in manufacturing for material processing, medicine, microelectronics, emission studies, lyophylisation, freeze drying and packaging. The quickly developing field of vacuum mechatronics will also be the driving force for the realization of an advanced era of totally enclosed clean manufacturing cells. High technology manufacturing has increasingly demanding requirements for precision manipulation, in situ process monitoring and contamination-free environments. To remove the contamination problems associated with human workers, the tendency in many manufacturing processes is to move towards total automation. This will become a requirement in the near future for e.g., microelectronics manufacturing. Automation in ultra-clean manufacturing environments is evolving into the concept of self-contained and fully enclosed manufacturing. A Self Contained Automated Robotic Factory (SCARF) is being developed as a flexible research facility for totally enclosed manufacturing. The construction and successful operation of a SCARF will provide a novel, flexible, self-contained, clean, vacuum manufacturing environment. SCARF also requires very high reliability and intelligent control. The trends in vacuum mechatronics and some of the key research issues are reviewed.

Moisture Adsorption Isotherm and Storability of Hazelnut Inshells and Kernels Produced in Oregon, USA.

PubMed

Jung, Jooyeoun; Wang, Wenjie; McGorrin, Robert J; Zhao, Yanyun

2018-02-01

Moisture adsorption isotherms and storability of dried hazelnut inshells and kernels produced in Oregon were evaluated and compared among cultivars, including Barcelona, Yamhill, and Jefferson. Experimental moisture adsorption data fitted to Guggenheim-Anderson-de Boer (GAB) model, showing less hygroscopic properties in Yamhill than other cultivars of inshells and kernels due to lower content of carbohydrate and protein, but higher content of fat. The safe levels of moisture content (MC, dry basis) of dried inshells and kernels for reaching kernel water activity (a w ) ≤0.65 were estimated using the GAB model as 11.3% and 5.0% for Barcelona, 9.4% and 4.2% for Yamhill, and 10.7% and 4.9% for Jefferson, respectively. Storage conditions (2 °C at 85% to 95% relative humidity [RH], 10 °C at 65% to 75% RH, and 27 °C at 35% to 45% RH), times (0, 4, 8, or 12 mo), and packaging methods (atmosphere vs. vacuum) affected MC, a w , bioactive compounds, lipid oxidation, and enzyme activity of dried hazelnut inshells or kernels. For inshells packaged at woven polypropylene bag, MC and a w of inshells and kernels (inside shells) increased at 2 and 10 °C, but decreased at 27 °C during storage. For kernels, lipid oxidation and polyphenol oxidase activity also increased with extended storage time (P < 0.05), and MC and a w of vacuum packaged samples were more stable during storage than those atmospherically packaged ones. Principal component analysis showed correlation of kernel qualities with storage condition, time, and packaging method. This study demonstrated that the ideal storage condition or packaging method varied among cultivars due to their different moisture adsorption and physicochemical and enzymatic stability during storage. Moisture adsorption isotherm of hazelnut inshells and kernels is useful for predicting the storability of nuts. This study found that water adsorption and storability varied among the different cultivars of nuts, in which Yamhill was less hygroscopic than Barcelona and Jefferson, thus more stable during storage. For ensuring food safety and quality of nuts during storage, each cultivar of kernels should be dried to a certain level of MC. Lipid oxidation and enzyme activity of kernel could be increased with extended storage time. Vacuum packaging was recommended to kernels for reducing moisture adsorption during storage. © 2018 Institute of Food Technologists®.

Long-Term Stability of NIST Chip-Scale Atomic Clock Physics Packages

DTIC Science & Technology

2007-01-01

vacuum packaging), as has been demonstrated by Lutwak et al. [3]. Nevertheless, we tried to investigate the causes for the frequency shifts of...stability,” Optics Express, 13, 1249-1253. [3] R. Lutwak , J. Deng, W. Riley, M. Varghese, J. Leblanc, G. Tepolt, M. Mescher, D. K. Serkland, K. M. Geib...the 1st Annual Multiconference on Electronics and Photonics, 7-11 November 2006, Guanajuato, Mexico, in press. [6] R. Lutwak , P. Vlitas, M

Vacuum foil insulation system

DOEpatents

Hanson, John P.; Sabolcik, Rudolph E.; Svedberg, Robert C.

1976-11-16

In a multifoil thermal insulation package having a plurality of concentric cylindrical cups, means are provided for reducing heat loss from the penetration region which extends through the cups. At least one cup includes an integral skirt extending from one end of the cup to intersection with the penetration means. Assembly of the insulation package with the skirted cup is facilitated by splitting the cup to allow it to be opened up and fitted around the other cups during assembly.

Innovative on-chip packaging applied to uncooled IRFPA

NASA Astrophysics Data System (ADS)

Dumont, Geoffroy; Arnaud, Agnès; Imperinetti, Pierre; Mottin, Eric; Simoens, François; Vialle, Claire; Rabaud, Wilfried; Grand, Gilles; Baclet, Nathalie

2008-03-01

The Laboratoire Infrarouge (LIR) of the Laboratoire d'Electronique et de Technologie de l'Information (LETI) has been involved in the development of microbolometers for over fifteen years. Two generations of technology have been transferred to ULIS and LETI is still working to improve performances of low cost detectors. Simultaneously, packaging still represents a significant part of detectors price. Reducing production costs would contribute to keep on extending applications of uncooled IRFPA to high volume markets like automotive. Therefore LETI develops an onchip packaging technology dedicated to microbolometers. The efficiency of a micropackaging technology for microbolometers relies on two major technical specifications. First, it must include an optical window with a high transmittance for the IR band, so as to maximize the detector absorption. Secondly, in order to preserve the thermal insulation of the detector, the micropackaging must be hermetically closed to maintain a vacuum level lower than 10 -3mbar. This paper presents an original microcap structure that enables the use of IR window materials as sealing layers to maintain the expected vacuum level. The modelling and integration of an IR window suitable for this structure is also presented. This zero level packaging technology is performed in a standard collective way, in continuation of bolometers' technology. The CEA-LETI, MINATEC presents status of these developments concerning this innovating technology including optical simulations results and SEM views of technical realizations.

Protection of microelectronic devices during packaging

DOEpatents

Peterson, Kenneth A.; Conley, William R.

2002-01-01

The present invention relates to a method of protecting a microelectronic device during device packaging, including the steps of applying a water-insoluble, protective coating to a sensitive area on the device; performing at least one packaging step; and then substantially removing the protective coating, preferably by dry plasma etching. The sensitive area can include a released MEMS element. The microelectronic device can be disposed on a wafer. The protective coating can be a vacuum vapor-deposited parylene polymer, silicon nitride, metal (e.g. aluminum or tungsten), a vapor deposited organic material, cynoacrylate, a carbon film, a self-assembled monolayered material, perfluoropolyether, hexamethyldisilazane, or perfluorodecanoic carboxylic acid, silicon dioxide, silicate glass, or combinations thereof. The present invention also relates to a method of packaging a microelectronic device, including: providing a microelectronic device having a sensitive area; applying a water-insoluble, protective coating to the sensitive area; providing a package; attaching the device to the package; electrically interconnecting the device to the package; and substantially removing the protective coating from the sensitive area.

Temporary coatings for protection of microelectronic devices during packaging

DOEpatents

Peterson, Kenneth A.; Conley, William R.

2005-01-18

The present invention relates to a method of protecting a microelectronic device during device packaging, including the steps of applying a water-insoluble, temporary protective coating to a sensitive area on the device; performing at least one packaging step; and then substantially removing the protective coating, preferably by dry plasma etching. The sensitive area can include a released MEMS element. The microelectronic device can be disposed on a wafer. The protective coating can be a vacuum vapor-deposited parylene polymer, silicon nitride, metal (e.g. aluminum or tungsten), a vapor deposited organic material, cynoacrylate, a carbon film, a self-assembled monolayered material, perfluoropolyether, hexamethyldisilazane, or perfluorodecanoic carboxylic acid, silicon dioxide, silicate glass, or combinations thereof. The present invention also relates to a method of packaging a microelectronic device, including: providing a microelectronic device having a sensitive area; applying a water-insoluble, protective coating to the sensitive area; providing a package; attaching the device to the package; electrically interconnecting the device to the package; and substantially removing the protective coating from the sensitive area.

Development of shelf stable pork sausages using hurdle technology and their quality at ambient temperature (37±1°C) storage.

PubMed

Thomas, R; Anjaneyulu, A S R; Kondaiah, N

2008-05-01

Shelf stable pork sausages were developed using hurdle technology and their quality was evaluated during ambient temperature (37±1°C) storage. Hurdles incorporated were low pH, low water activity, vacuum packaging and post package reheating. Dipping in potassium sorbate solution prior to vacuum packaging was also studied. Reheating increased the pH of the sausages by 0.17units as against 0.11units in controls. Incorporation of hurdles significantly decreased emulsion stability, cooking yield, moisture and fat percent, yellowness and hardness, while increasing the protein percent and redness. Hurdle treatment reduced quality deterioration during storage as indicated by pH, TBARS and tyrosine values. About 1 log reduction in total plate count was observed with the different hurdles as were reductions in the coliform, anaerobic, lactobacilli and Staphylococcus aureus counts. pH, a(w) and reheating hurdles inhibited yeast and mold growth up to day 3, while additional dipping in 1% potassium sorbate solution inhibited their growth throughout the 9 days storage. Despite low initial sensory appeal, the hurdle treated sausages had an overall acceptability in the range 'very good' to 'good' up to day 6.

Compact atom interferometer using single laser

NASA Astrophysics Data System (ADS)

Chiow, Sheng-Wey; Yu, Nan

2017-04-01

Atom interferometer (AI) based sensors exhibit precision and accuracy unattainable with classical sensors, thanks to the inherent stability of atomic properties. The complexity of required laser system and the size of vacuum chamber driven by optical access requirement limit the applicability of such technology in size, weight, and power (SWaP) challenging environments, such as in space. For instance, a typical physics package of AI includes six viewports for laser cooling and trapping, two for AI beams, and two more for detection and a vacuum pump. Similarly, a typical laser system for an AI includes two lasers for cooling and repumping, and two for Raman transitions as AI beam splitters. In this presentation, we report our efforts in developing a miniaturized atomic accelerometer for planetary exploration. We will describe a physics package configuration having minimum optical access (thus small volume), and a laser and optics system utilizing a single laser for the sensor operation. Preliminary results on acceleration sensitivity will be discussed. We will also illustrate a path for further packaging and integration based on the demonstrated concepts. This research was carried out at the Jet Propulsion Laboratory, California Institute of Technology, under a contract with the National Aeronautics and Space Administration.

Wafer-to-wafer bonding of nonplanarized MEMS surfaces using solder

NASA Astrophysics Data System (ADS)

Sparks, D.; Queen, G.; Weston, R.; Woodward, G.; Putty, M.; Jordan, L.; Zarabadi, S.; Jayakar, K.

2001-11-01

The fabrication and reliability of a solder wafer-to-wafer bonding process is discussed. Using a solder reflow process allows vacuum packaging to be accomplished with unplanarized complementary metal-oxide semiconductor (CMOS) surface topography. This capability enables standard CMOS processes, and integrated microelectromechanical systems devices to be packaged at the chip-level. Alloy variations give this process the ability to bond at lower temperatures than most alternatives. Factors affecting hermeticity, shorts, Q values, shifting cavity pressure, wafer saw cleanliness and corrosion resistance will be covered.

Engineering report on the OAO-2 Wisconsin experiment package

NASA Technical Reports Server (NTRS)

Bendell, C. B.

1972-01-01

The continued useful operation of the OAO-2 Wisconsin Experiment Package (WEP) for almost three years after its December 1968 launch is evidence of a superior engineering accomplishment. Reliability features of the experiment concept and design which have contributed to its long life are presented. Data anomalies and partial failures are summarized along with conclusions regarding their causes. The thermal, vacuum and radiation effects of the space environment are shown to be minimal and quite localized within the WEP.

Instrumental meat quality of veal calves reared under three management systems and color evolution of meat stored in three packaging systems.

PubMed

Ripoll, G; Albertí, P; Casasús, I; Blanco, M

2013-02-01

This study evaluated the meat quality of grazing suckling calves (GR), suckling plus supplemented calves (SUP) and weaned calves finished on concentrates (FIN) and the color evolution of meat packaged in film (FILM), modified atmosphere packaging (MAP) and vacuum packaging (VAC). Intramuscular fat was quite low for all treatments and GR meat had greater percentages of PUFA and lower SFA, MUFA and n6/n3 than SUP and FIN. FIN and SUP meat had more L* and was more tender when aged than GR. GR meat was tough and dark. The packaging system was more influential on meat color than the feeding management. VAC had the lowest values of metmyoglobin when aged. MAP had the greatest L* and hue angle and the lowest redness after 13 d of aging, thus MAP was the packaging with the shortest shelf life. Copyright © 2012 Elsevier Ltd. All rights reserved.

Controlling Blown Pack Spoilage Using Anti-Microbial Packaging

PubMed Central

Reid, Rachael; Tyuftin, Andrey A.; Kerry, Joe P.; Whyte, Paul; Bolton, Declan

2017-01-01

Active (anti-microbial) packaging was prepared using three different formulations; Auranta FV; Inbac-MDA and sodium octanoate at two concentrations (2.5 and 3.5 times their minimum inhibitory concentration (MIC, the lowest concentration that will inhibit the visible growth of the organisms) against Clostridium estertheticum, DSMZ 8809). Inoculated beef samples were packaged using the active packaging and monitored for 100 days storage at 2 °C for blown pack spoilage. The time to the onset of blown pack spoilage was significantly (p < 0.01) increased using Auranta FV and sodium octanoate (caprylic acid sodium salt) at both concentrations. Moreover, sodium octanoate packs had significantly (p < 0.01) delayed blown pack spoilage as compared to Auranta FV. It was therefore concluded that Auranta FV or sodium octanoate, incorporated into the packaging materials used for vacuum packaged beef, would inhibit blown pack spoilage and in the case of the latter, well beyond the 42 days storage period currently required for beef primals. PMID:28805679

Controlling Blown Pack Spoilage Using Anti-Microbial Packaging.

PubMed

Reid, Rachael; Bolton, Declan; Tiuftin, Andrey A; Kerry, Joe P; Fanning, Séamus; Whyte, Paul

2017-08-12

Active (anti-microbial) packaging was prepared using three different formulations; Auranta FV; Inbac-MDA and sodium octanoate at two concentrations (2.5 and 3.5 times their minimum inhibitory concentration (MIC, the lowest concentration that will inhibit the visible growth of the organisms) against Clostridium estertheticum , DSMZ 8809). Inoculated beef samples were packaged using the active packaging and monitored for 100 days storage at 2 °C for blown pack spoilage. The time to the onset of blown pack spoilage was significantly ( p < 0.01) increased using Auranta FV and sodium octanoate (caprylic acid sodium salt) at both concentrations. Moreover, sodium octanoate packs had significantly ( p < 0.01) delayed blown pack spoilage as compared to Auranta FV. It was therefore concluded that Auranta FV or sodium octanoate, incorporated into the packaging materials used for vacuum packaged beef, would inhibit blown pack spoilage and in the case of the latter, well beyond the 42 days storage period currently required for beef primals.

Study of cavity effect in micro-Pirani gauge chamber with improved sensitivity for high vacuum regime

NASA Astrophysics Data System (ADS)

Zhang, Guohe; Lai, Junhua; Kong, Yanmei; Jiao, Binbin; Yun, Shichang; Ye, Yuxin

2018-05-01

Ultra-low pressure application of Pirani gauge needs significant improvement of sensitivity and expansion of measureable low pressure limit. However, the performance of Pirani gauge in high vacuum regime remains critical concerns since gaseous thermal conduction with high percentage is essential requirement. In this work, the heat transfer mechanism of micro-Pirani gauge packaged in a non-hermetic chamber was investigated and analyzed compared with the one before wafer-level packaging. The cavity effect, extremely important for the efficient detection of low pressure, was numerically and experimentally analyzed considering the influence of the pressure, the temperature and the effective heat transfer area in micro-Pirani gauge chamber. The thermal conduction model is validated by experiment data of MEMS Pirani gauges with and without capping. It is found that nature gaseous convection in chamber, determined by the Rayleigh number, should be taken into consideration. The experiment and model calculated results show that thermal resistance increases in the molecule regime, and further increases after capping due to the suppression of gaseous convection. The gaseous thermal conduction accounts for an increasing percentage of thermal conduction at low pressure while little changes at high pressure after capping because of the existence of cavity effect improving the sensitivity of cavity-effect-influenced Pirani gauge for high vacuum regime.

MEMS packaging: state of the art and future trends

NASA Astrophysics Data System (ADS)

Bossche, Andre; Cotofana, Carmen V. B.; Mollinger, Jeff R.

1998-07-01

Now that the technology for Integrated sensor and MEMS devices has become sufficiently mature to allow mass production, it is expected that the prices of bare chips will drop dramatically. This means that the package prices will become a limiting factor in market penetration, unless low cost packaging solutions become available. This paper will discuss the developments in packaging technology. Both single-chip and multi-chip packaging solutions will be addressed. It first starts with a discussion on the different requirements that have to be met; both from a device point of view (open access paths to the environment, vacuum cavities, etc.) and from the application point of view (e.g. environmental hostility). Subsequently current technologies are judged on their applicability for MEMS and sensor packaging and a forecast is given for future trends. It is expected that the large majority of sensing devices will be applied in relative friendly environments for which plastic packages would suffice. Therefore, on the short term an important role is foreseen for recently developed plastic packaging techniques such as precision molding and precision dispensing. Just like in standard electronic packaging, complete wafer level packaging methods for sensing devices still have a long way to go before they can compete with the highly optimized and automated plastic packaging processes.

The application of carbon monoxide in meat packaging needs to be re-evaluated within the EU: An overview.

PubMed

Van Rooyen, Lauren Anne; Allen, Paul; O'Connor, David I

2017-10-01

Carbon monoxide (CO) has many value-added benefits in meat packaging due to its colour stabilising effects and enhancement of meat quality attributes. The regulation of CO within meat packaging varies worldwide and remains a topical and controversial issue. CO is prohibited in the EU for use in meat packaging mainly due to fears it may mask spoilage therefore misleading consumers. The issue of consumer acceptance of CO was not considered. This article reviews the most pertinent literature to assess if the problems associated with the prohibition have been addressed. Applying CO pretreatments prior to vacuum packaging enhances colour while allowing discolouration to occur by the use-by-date, thereby addressing concerns about safety. Recent work showing European consumer acceptance of CO in meat packaging demonstrates its future potential within the EU. The information provided may support framing future policies intended to assure consumer protection, safety, choice and interest. Re-evaluation of permitting CO as a packaging gas within the EU may be warranted. Copyright © 2017 Elsevier Ltd. All rights reserved.

Effect of Rosmarinus officinalis L. essential oil combined with different packaging conditions to extend the shelf life of refrigerated beef meat.

PubMed

Sirocchi, Veronica; Devlieghere, Frank; Peelman, Nanou; Sagratini, Gianni; Maggi, Filippo; Vittori, Sauro; Ragaert, Peter

2017-04-15

Rosemary essential oil (REO) contains bioactives having antioxidant and antimicrobial properties. This work investigated the effect of REO combined with modified atmosphere packaging conditions (MAP), in our case, aerobic, vacuum or high O 2 , to extend the shelf life of beef. Beef slices were wrapped in special three-layer sheets of packaging material, some with a coating of REO (active packaging, AP), and some without REO (non active packaging, NAP), and stored at 4°C for 20days. The use of REO proved efficacious in every storage condition, as seen in the lower counts of psychrotrophics, Brochothrix thermosphacta, Pseudomonas spp., and Enterobacteriaceae in AP meat compared to NAP meat. Sensory and colourimetric analyses showed that the best packaging conditions were high-O 2 atmosphere in combination with REO. Based on microbiological data, shelf life of beef was 5-6days for AP samples packaged under aerobic conditions and 14-15days for AP samples in high-O 2 conditions. Copyright © 2016 Elsevier Ltd. All rights reserved.

Effectiveness of antimicrobial food packaging materials.

PubMed

Cooksey, K

2005-10-01

Antimicrobial additives have been used successfully for many years as direct food additives. The literature provides evidence that some of these additives may be effective as indirect food additives incorporated into food packaging materials. Antimicrobial food packaging is directed toward the reduction of surface contamination of processed, prepared foods such as sliced meats and Frankfurter sausages (hot dogs). The use of such packaging materials is not meant to be a substitute for good sanitation practices, but it should enhance the safety of food as an additional hurdle for the growth of pathogenic and/or spoilage microorganisms. Studies have focused on establishing methods for coating low-density polyethylene film or barrier films with methyl cellulose as a carrier for nisin. These films have significantly reduced the presence of Listeria monocytogenes in solutions and in vacuum packaged hot dogs. Other research has focused on the use of chitosan to inhibit L. monocytogenes and chlorine dioxide sachets for the reduction of Salmonella on modified atmosphere-packaged fresh chicken breasts. Overall, antimicrobial packaging shows promise as an effective method for the inhibition of certain bacteria in foods, but barriers to their commercial implementation continue to exist.

Effects of diet, packaging, and irradiation on protein oxidation, lipid oxidation, and color of raw broiler thigh meat during refrigerated storage.

PubMed

Xiao, S; Zhang, W G; Lee, E J; Ma, C W; Ahn, D U

2011-06-01

This study was designed to evaluate the effects of dietary treatment, packaging, and irradiation singly or in combination on the oxidative stability of broiler chicken thigh meat. A total of 120 four-week-old chickens were divided into 12 pens (10 birds/pen), and 4 pens of broilers were randomly assigned to a control oxidized diet (5% oxidized oil) or an antioxidant-added diet [500 IU of vitamin E + 200 mg/kg of butylated hydroxyanisole (BHA)] and fed for 2 wk. After slaughter, thigh meats were separated, ground, packaged in either oxygen-permeable or oxygen-impermeable vacuum bags, and irradiated at 0 or 3 kGy. Lipid oxidation (TBA-reactive substances), protein oxidation (carbonyl), and color of the meat were measured at 1, 4, and 7 d of refrigerated storage. The lipid and protein oxidation of thigh meats from birds fed the diet supplemented with antioxidants (vitamin E + BHA) was significantly lower than the lipid and protein oxidation of birds fed the control diet, whereas the lipid and protein oxidation of broilers fed the oxidized oil diet was higher than that of birds fed the control diet. Vacuum packaging slowed, but irradiation accelerated, the lipid and protein oxidation of thigh meat during storage. Dietary antioxidants (vitamin E + BHA) and irradiation treatments showed a stronger effect on lipid oxidation than on protein oxidation. A significant correlation between lipid and protein oxidation in meat was found during storage. Dietary supplementation of vitamin E + BHA and the irradiation treatment increased the lightness and redness of thigh meat, respectively. It is suggested that appropriate use of dietary antioxidants in combination with packaging could be effective in minimizing oxidative changes in irradiated raw chicken thigh meat.

Thermally Stable Gold Nanoparticles with a Crosslinked Diblock Copolymer Shell

NASA Astrophysics Data System (ADS)

Jang, Se Gyu; Khan, Anzar; Hawker, Craig J.; Kramer, Edward J.

2010-03-01

The use of polymer-coated Au nanoparticles prepared using oligomeric- or polymeric-ligands tethered by Au-S bonds for incorporation into block copolymer templates under thermal processing has been limited due to dissociation of the Au-S bond at T > 100^oC where compromises their colloidal stability. We report a simple route to prepare sub-5nm gold nanoparticles with a thermally stable polymeric shell. An end-functional thiol ligand consisting of poly(styrene-b-1,2&3,4-isoprene-SH) is synthesized by anionic polymerization. After a standard thiol ligand synthesis of Au nanoparticles, the inner PI block is cross-linked through reaction with 1,1,3,3-tetramethyldisiloxane. Gold nanoparticles with the cross-linked shell are stable in organic solvents at 160^oC as well as in block copolymer films of PS-b-P2VP annealed in vacuum at 170^oC for several days. These nanoparticles can be designed to strongly segregate to the PS-P2VP interface resulting in very large Au nanoparticle volume fractions φp without macrophase separation as well as transitions between lamellar and bicontinuous morphologies as φp increases.

The effect of temperature during retail display on the colour stability of CO pretreated vacuum packaged beef steaks.

PubMed

Van Rooyen, Lauren Anne; Allen, Paul; Gallagher, Eimear I; O'Connor, David I

2018-05-24

The effect of CO pretreatments applied to beef striploin steaks (Longissimus thoracis et lumborum, LTL) prior to vacuum packaging and display temperature on colour stability, shelf life and tenderness was determined. Steaks were exposed to 5% CO, 60% CO 2 and 35% N 2 for 3 (CO3), 5 (CO5) or 7 (CO7) h, followed by 28 days display at 2 °C (good industry practice) or 6 °C (mild abuse). CO5 was the optimum exposure time as it induced the desirable colour while not retaining the bright colour, irrespective of display temperature. K/S ratios confirmed that CO pretreatment did not mask spoilage and could be more sensitive than colour parameters at monitoring discoloration as colour was not retained. Exposure to CO did not have any negative effect on meat quality attributes, while mild temperature abuse (6 °C) increased purge loss and decreased pH. Copyright © 2018 Elsevier Ltd. All rights reserved.

Latest improvements in microbolometer thin film packaging: paving the way for low-cost consumer applications

NASA Astrophysics Data System (ADS)

Yon, J. J.; Dumont, G.; Goudon, V.; Becker, S.; Arnaud, A.; Cortial, S.; Tisse, C. L.

2014-06-01

Silicon-based vacuum packaging is a key enabling technology for achieving affordable uncooled Infrared Focal Plane Arrays (IRFPA) required by a promising mass market that shows momentum for some extensive consumer applications, such as automotive driving assistance, smart presence localization and building management. Among the various approaches studied worldwide, CEA, LETI in partnership with ULIS is committed to the development of a unique technology referred to as PLP (Pixel Level Packaging). In this PLP technology, each bolometer pixel is sealed under vacuum using a transparent thin film deposition on wafer. PLP operates as an array of hermetic micro caps above the focal plane, each enclosing a single microbolometer. In continuation of our on-going studies on PLP for regular QVGA IRFPAs, this paper emphasizes on the innate scalability of the technology which was successfully demonstrated through the development of an 80 × 80 pixel IRFPA. The relevance of the technology with regard to the two formats is discussed, considering both performance and cost issues. We show that the suboptimal fill factor inherent to the PLP arrangement is not so critical when considering smaller arrays preferably fitted for consumer applications. The discussion is supported with the electro-optical performance measurements of the PLP-based 80×80 demonstrator.

Industrial application of an antilisterial strain of Lactobacillus sakei as a protective culture and its effect on the sensory acceptability of cooked, sliced, vacuum-packaged meats.

PubMed

Bredholt, S; Nesbakken, T; Holck, A

2001-06-15

The application of a protective lactic acid bacterium (LAB) during the commercial production of cooked meat products is described. The LAB, a strain of Lactobacillus sakei, was previously isolated from cooked ham and inhibited growth of Listeria monocytogenes and Escherichia coli O157:H7 in this product. L. sakei was applied to the cooked products at a concentration of 10(5)-10(6) cfu/g immediately before slicing and vacuum-packaging using a hand-operated spraying bottle. The LAB strain inhibited growth of 10(3) cfu/g of a cocktail of three rifampicin resistant mutant L. monocytogenes strains both at 8 degrees C and 4 degrees C. Consumer acceptance tests of cooked ham and of servelat sausage, a Norwegian non-fermented cooked meat sausage, showed that control and inoculated products were equally acceptable. The products were still acceptable after storage for 28 days at 4 degrees C and, after opening the packages, for a further 5 days at 4 degrees C. The findings presented here confirm that the L. sakei strain is suitable for use as a protective culture and may technically easily be implemented in the commercial production of cooked meat products.

Thermal Interface Materials Selection and Application Guidelines: In Perspective of Xilinx Virtex-5QV Thermal Management

NASA Technical Reports Server (NTRS)

Suh, Jong-ook; Dillon, R. Peter; Tseng, Stephen

2015-01-01

The heat from high-power microdevices for space, such as Xilinx Virtex 4 and 5 (V4 and V5), has to be removed mainly through conduction in the space vacuum environment. The class-Y type packages are designed to remove the heat from the top of the package, and the most effective method to remove heat from the class-Y type packages is to attach a heat transfer device on the lid of the package and to transfer the heat to frame or chassis. When a heat transfer device is attached to the package lid, the surfaces roughness of the package lid and the heat transfer device reduces the effective contact area between the two. The reduced contact area results in increased thermal contact resistance, and a thermal interface material is required to reduce the thermal contact resistance by filling in the gap between the surfaces of the package lid and the heat transfer device. The current report describes JPL's FY14 NEPP task study on property requirements of TIM and impact of TIM properties on the packaging reliability. The current task also developed appratuses to investigate the performances of TIMs in the actual mission environment.

Effect of electron irradiation and packaging atmosphere on the survival of aeromonas hydrophila in minced poultry meat

NASA Astrophysics Data System (ADS)

Stecchini, M. L.; Sarais, I.; Del Torre, M.; Fuochi, P. G.

1995-02-01

Resistance to electron irradiation of Aeromonas hydrophila inoculated in minced poultry meat packed in presence of air or under vacuum was examined. Surviving bacteria were counted on starch ampicillin agar containing 100 μg/ml of ampicillin. Radiation resistance, expressed as D 10 values, was calculated from the survival curves and found to be 0.12 and 0.12 kGy in poultry meat packed in air or under vacuum respectively. Storage at 2°C of meat samples irradiated at 0.5 kGy further reduced the number of A. hydrophila.

Effect of High Temperature Storage in Vacuum, Air, and Humid Conditions on Degradation of Gold/Aluminum Wire Bonds in PEMs

NASA Technical Reports Server (NTRS)

Teverovsky, Alexander

2006-01-01

Microcircuits encapsulated in three plastic package styles were stored in different environments at temperatures varying from 130 C to 225 C for up to 4,000 hours in some cases. To assess the effect of oxygen, the parts were aged at high temperatures in air and in vacuum chambers. The effect of humidity was evaluated during long-term highly accelerated temperature and humidity stress testing (HAST) at temperatures of 130 C and 150 C. High temperature storage testing of decapsulated microcircuits in air, vacuum, and HAST chambers was carried out to evaluate the role of molding compounds in the environmentally-induced degradation and failure of wire bonds (WB). This paper reports on accelerating factors of environment and molding compound on WB failures. It has been shown that all environments, including oxygen, moisture, and the presence of molding compounds reduce time-to-failures compared to unencapsulated devices in vacuum conditions. The mechanism of the environmental effect on KB degradation is discussed.

Meat Processing Plant Microbiome and Contamination Patterns of Cold-Tolerant Bacteria Causing Food Safety and Spoilage Risks in the Manufacture of Vacuum-Packaged Cooked Sausages

PubMed Central

Rahkila, Riitta; Ali, Javeria; Rousu, Juho; Björkroth, K. Johanna

2015-01-01

Refrigerated food processing facilities are specific man-made niches likely to harbor cold-tolerant bacteria. To characterize this type of microbiota and study the link between processing plant and product microbiomes, we followed and compared microbiota associated with the raw materials and processing stages of a vacuum-packaged, cooked sausage product affected by a prolonged quality fluctuation with occasional spoilage manifestations during shelf life. A total of 195 samples were subjected to culturing and amplicon sequence analyses. Abundant mesophilic psychrotrophs were detected within the microbiomes throughout the different compartments of the production plant environment. However, each of the main genera of food safety and quality interest, e.g., Leuconostoc, Brochothrix, and Yersinia, had their own characteristic patterns of contamination. Bacteria from the genus Leuconostoc, commonly causing spoilage of cold-stored, modified-atmosphere-packaged foods, were detected in high abundance (up to >98%) in the sausages studied. The same operational taxonomic units (OTUs) were, however, detected in lower abundances in raw meat and emulsion (average relative abundance of 2% ± 5%), as well as on the processing plant surfaces (<4%). A completely different abundance profile was found for OTUs phylogenetically close to the species Yersinia pseudotuberculosis. These OTUs were detected in high abundance (up to 28%) on the processing plant surfaces but to a lesser extent (<1%) in raw meat, sausage emulsion, and sausages. The fact that Yersinia-like OTUs were found on the surfaces of a high-hygiene packaging compartment raises food safety concerns related to their resilient existence on surfaces. PMID:26231646

Thermotolerance of meat spoilage lactic acid bacteria and their inactivation in vacuum-packaged vienna sausages.

PubMed

Franz, C M; von Holy, A

1996-02-01

Heat resistance of three meat spoilage lactic acid bacteria was determined in vitro. D-values at 57, 60 and 63 degrees C were 52.9, 39.3 and 32.5 s for Lactobacillus sake, 34.9, 31.3 and 20.2 s for Leuconostoc mesenteroides and 22.5, 15.6 and 14.4 s for Lactobacillus curvatus, respectively. The three lactic acid bacteria were heat sensitive, as one log reductions in numbers were achieved at 57 degrees C in less than 60 s. Z-values could not be accurately determined as D-values did not change by a factor of 10 over the temperature range studied. In-package pasteurization processes were calculated using the highest in vitro D-value and applied to vacuum-packaged vienna sausages. Microbiological shelf life (time for lactic acid bacteria count to reach 5 x 10(6) CFU/g) increased from 7 days for non-pasteurized samples to 67, 99 and 119 days for samples of the three pasteurization treatments at 8 degrees C storage. Enterobacteriaceae were detected at levels of log 4.0 CFU/g in non-pasteurized samples, but were reduced to < log 1.0 CFU/g in pasteurized samples. The incidence of listeriae in non-pasteurized samples was low as only one Listeria innocua strain was isolated. No Listeria spp. were isolated from pasteurized samples. Numbers of Clostridium isolates increased from one in non-pasteurized samples to 25 in pasteurized samples. Increasing incidences of clostridia, and the presence of C. perfringens in pasteurized samples indicated that in-package pasteurization could compromise product safety.

Combined effect of cooking (grilling and roasting) and chilling storage (with and without air) on lipid and cholesterol oxidation in chicken breast.

PubMed

Conchillo, Ana; Ansorena, Diana; Astiasarán, Iciar

2003-05-01

The oxidation of the lipid fraction and cholesterol in raw and cooked chicken breast samples stored for 0 and 6 days at 4 degrees C under aerobic conditions and in vacuum packaging was studied. The multivariate statistical analysis showed significant effects of both culinary process and storage conditions on the lipid and cholesterol oxidation process, with a significant interaction between the two variables. Aerobic storage increased thiobarbituric acid reactive substances (TBA) from 0.04 to 0.06 ppm for raw samples, from 0.21 to 1.20 ppm for grilled samples, and from 0.24 to 1.62 ppm for roasted samples. During vacuum storage, only roasted samples showed significant increases in TBA. Levels of total cholesterol oxidation products (COP) remained low (2.88 to 4.35 microg/g of lipid) for all raw samples. Cooking increased COP levels to 12.85 and 11.54 microg/ g of lipid for grilled and roasted samples, respectively. Total COP and all individual COP except for cholestanetriol were significantly correlated with TBA and the peroxide index. However, the most extensive effect was attributable to the aerobic storage of cooked samples, which led to COP levels of 92.35 and 88.60 microg/g of lipid in grilled and roasted samples, respectively. Vacuum packaging did not increase COP levels for cooked samples.

Single-Particle Detection of Transcription following Rotavirus Entry

PubMed Central

Salgado, Eric N.; Upadhyayula, Srigokul

2017-01-01

ABSTRACT Infectious rotavirus particles are triple-layered, icosahedral assemblies. The outer layer proteins, VP4 (cleaved to VP8* and VP5*) and VP7, surround a transcriptionally competent, double-layer particle (DLP), which they deliver into the cytosol. During entry of rhesus rotavirus, VP8* interacts with cell surface gangliosides, allowing engulfment into a membrane vesicle by a clathrin-independent process. Escape into the cytosol and outer-layer shedding depend on interaction of a hydrophobic surface on VP5* with the membrane bilayer and on a large-scale conformational change. We report here experiments that detect the fate of released DLPs and their efficiency in initiating RNA synthesis. By replacing the outer layer with fluorescently tagged, recombinant proteins and also tagging the DLP, we distinguished particles that have lost their outer layer and entered the cytosol (uncoated) from those still within membrane vesicles. We used fluorescent in situ hybridization with probes for nascent transcripts to determine how soon after uncoating transcription began and what fraction of the uncoated particles were active in initiating RNA synthesis. We detected RNA synthesis by uncoated particles as early as 15 min after adding virus. The uncoating efficiency was 20 to 50%; of the uncoated particles, about 10 to 15% synthesized detectable RNA. In the format of our experiments, about 10% of the added particles attached to the cell surface, giving an overall ratio of added particles to RNA-synthesizing particles of between 250:1 and 500:1, in good agreement with the ratio of particles to focus-forming units determined by infectivity assays. Thus, RNA synthesis by even a single, uncoated particle can initiate infection in a cell. IMPORTANCE The pathways by which a virus enters a cell transform its packaged genome into an active one. Contemporary fluorescence microscopy can detect individual virus particles as they enter cells, allowing us to map their multistep entry pathways. Rotaviruses, like most viruses that lack membranes of their own, disrupt or perforate the intracellular, membrane-enclosed compartment into which they become engulfed following attachment to a cell surface, in order to gain access to the cell interior. The properties of rotavirus particles make it possible to determine molecular mechanisms for these entry steps. In the work described here, we have asked the following question: what fraction of the rotavirus particles that penetrate into the cell make new viral RNA? We find that of the cell-attached particles, between 20 and 50% ultimately penetrate, and of these, about 10% make RNA. RNA synthesis by even a single virus particle can initiate a productive infection. PMID:28701394

Modulation of a Pore in the Capsid of JC Polyomavirus Reduces Infectivity and Prevents Exposure of the Minor Capsid Proteins

PubMed Central

Nelson, Christian D. S.; Ströh, Luisa J.; Gee, Gretchen V.; O'Hara, Bethany A.; Stehle, Thilo

2015-01-01

ABSTRACT JC polyomavirus (JCPyV) infection of immunocompromised individuals results in the fatal demyelinating disease progressive multifocal leukoencephalopathy (PML). The viral capsid of JCPyV is composed primarily of the major capsid protein virus protein 1 (VP1), and pentameric arrangement of VP1 monomers results in the formation of a pore at the 5-fold axis of symmetry. While the presence of this pore is conserved among polyomaviruses, its functional role in infection or assembly is unknown. Here, we investigate the role of the 5-fold pore in assembly and infection of JCPyV by generating a panel of mutant viruses containing amino acid substitutions of the residues lining this pore. Multicycle growth assays demonstrated that the fitness of all mutants was reduced compared to that of the wild-type virus. Bacterial expression of VP1 pentamers containing substitutions to residues lining the 5-fold pore did not affect pentamer assembly or prevent association with the VP2 minor capsid protein. The X-ray crystal structures of selected pore mutants contained subtle changes to the 5-fold pore, and no other changes to VP1 were observed. Pore mutant pseudoviruses were not deficient in assembly, packaging of the minor capsid proteins, or binding to cells or in transport to the host cell endoplasmic reticulum. Instead, these mutant viruses were unable to expose VP2 upon arrival to the endoplasmic reticulum, a step that is critical for infection. This study demonstrated that the 5-fold pore is an important structural feature of JCPyV and that minor modifications to this structure have significant impacts on infectious entry. IMPORTANCE JCPyV is an important human pathogen that causes a severe neurological disease in immunocompromised individuals. While the high-resolution X-ray structure of the major capsid protein of JCPyV has been solved, the importance of a major structural feature of the capsid, the 5-fold pore, remains poorly understood. This pore is conserved across polyomaviruses and suggests either that these viruses have limited structural plasticity in this region or that this pore is important in infection or assembly. Using a structure-guided mutational approach, we showed that modulation of this pore severely inhibits JCPyV infection. These mutants do not appear deficient in assembly or early steps in infectious entry and are instead reduced in their ability to expose a minor capsid protein in the host cell endoplasmic reticulum. Our work demonstrates that the 5-fold pore is an important structural feature for JCPyV. PMID:25609820

Expression and purification of recombinant polyomavirus VP2 protein and its interactions with polyomavirus proteins

NASA Technical Reports Server (NTRS)

Cai, X.; Chang, D.; Rottinghaus, S.; Consigli, R. A.; Spooner, B. S. (Principal Investigator)

1994-01-01

Recombinant polyomavirus VP2 protein was expressed in Escherichia coli (RK1448), using the recombinant expression system pFPYV2. Recombinant VP2 was purified to near homogeneity by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, electroelution, and Extracti-Gel chromatography. Polyclonal serum to this protein which reacted specifically with recombinant VP2 as well as polyomavirus virion VP2 and VP3 on Western blots (immunoblots) was produced. Purified VP2 was used to establish an in vitro protein-protein interaction assay with polyomavirus structural proteins and purified recombinant VP1. Recombinant VP2 interacted with recombinant VP1, virion VP1, and the four virion histones. Recombinant VP1 coimmunoprecipitated with recombinant VP2 or truncated VP2 (delta C12VP2), which lacked the carboxy-terminal 12 amino acids. These experiments confirmed the interaction between VP1 and VP2 and revealed that the carboxyterminal 12 amino acids of VP2 and VP3 were not necessary for formation of this interaction. In vivo VP1-VP2 interaction study accomplished by cotransfection of COS-7 cells with VP2 and truncated VP1 (delta N11VP1) lacking the nuclear localization signal demonstrated that VP2 was capable of translocating delta N11VP1 into the nucleus. These studies suggest that complexes of VP1 and VP2 may be formed in the cytoplasm and cotransported to the nucleus for virion assembly to occur.

Detection of secondary and backscattered electrons for 3D imaging with multi-detector method in VP/ESEM.

PubMed

Slówko, Witold; Wiatrowski, Artur; Krysztof, Michał

2018-01-01

The paper considers some major problems of adapting the multi-detector method for three-dimensional (3D) imaging of wet bio-medical samples in Variable Pressure/Environmental Scanning Electron Microscope (VP/ESEM). The described method pertains to "single-view techniques", which to create the 3D surface model utilise a sequence of 2D SEM images captured from a single view point (along the electron beam axis) but illuminated from four directions. The basis of the method and requirements resulting from them are given for the detector systems of secondary (SE) and backscattered electrons (BSE), as well as designs of the systems which could work in variable conditions. The problems of SE detection with application of the Pressure Limiting Aperture (PLA) as the signal collector are discussed with respect to secondary electron backscattering by a gaseous environment. However, the authors' attention is turned mainly to the directional BSE detection, realized in two ways. The high take off angle BSE were captured through PLA with use of the quadruple semiconductor detector placed inside the intermediate chamber, while BSE starting at lower angles were detected by the four-folded ionization device working in the sample chamber environment. The latter relied on a conversion of highly energetic BSE into low energetic SE generated on walls and a gaseous environment of the deep discharge gap oriented along the BSE velocity direction. The converted BSE signal was amplified in an ionising avalanche developed in the electric field arranged transversally to the gap. The detector system operation is illustrated with numerous computer simulations and examples of experiments and 3D images. The latter were conducted in a JSM 840 microscope with its combined detector-vacuum equipment which could extend capabilities of this high vacuum instrument toward elevated pressures (over 1kPa) and environmental conditions. Copyright © 2017 Elsevier Ltd. All rights reserved.

An evaluation of the effectiveness of FreshCase technology to extend the storage life of whole-muscle pork and ground pork sausage.

PubMed

Yang, X; Woerner, D R; McCullough, K R; Hasty, J D; Geornaras, I; Smith, G C; Sofos, J N; Belk, K E

2016-11-01

The objective of this study was to identify the maximum time of refrigerated storage before aerobic psychrotrophic bacteria grew to a level indicative of spoilage (7 log cfu/g) or other indicators of spoilage were observed for whole-muscle pork and ground pork sausage packaged using FreshCase technology. Pork chops and pork sausage were packaged using conventional vacuum packaging without nitrite in film (Control) or using FreshCase technology and were compared with respect to microbial counts, pH, instrumental color measurements, lipid oxidation level, and sensory properties. The storage life was 45 d for pork chops stored in FreshCase packages at 1°C and 19 d for ground pork sausage stored under the same condition. Results indicated that both pork chops and sausage stored in FreshCase packages retained redder color ( < 0.05) than those stored in Control packages. No differences ( > 0.05) existed between Control and FreshCase packaged samples for any off-odor detection for either pork chops or sausage. Moreover, levels of oxidative rancidity in all packages had low thiobarbituric acid reactive substances values. The results indicated that FreshCase technology can be used to extend storage life of pork products without having adverse effects on pork quality.

Wafer-level vacuum packaged resonant micro-scanning mirrors for compact laser projection displays

NASA Astrophysics Data System (ADS)

Hofmann, Ulrich; Oldsen, Marten; Quenzer, Hans-Joachim; Janes, Joachim; Heller, Martin; Weiss, Manfred; Fakas, Georgios; Ratzmann, Lars; Marchetti, Eleonora; D'Ascoli, Francesco; Melani, Massimiliano; Bacciarelli, Luca; Volpi, Emilio; Battini, Francesco; Mostardini, Luca; Sechi, Francesco; De Marinis, Marco; Wagner, Bernd

2008-02-01

Scanning laser projection using resonant actuated MEMS scanning mirrors is expected to overcome the current limitation of small display size of mobile devices like cell phones, digital cameras and PDAs. Recent progress in the development of compact modulated RGB laser sources enables to set up very small laser projection systems that become attractive not only for consumer products but also for automotive applications like head-up and dash-board displays. Within the last years continuous progress was made in increasing MEMS scanner performance. However, only little is reported on how mass-produceability of these devices and stable functionality even under harsh environmental conditions can be guaranteed. Automotive application requires stable MEMS scanner operation over a wide temperature range from -40° to +85°Celsius. Therefore, hermetic packaging of electrostatically actuated MEMS scanning mirrors becomes essential to protect the sensitive device against particle contamination and condensing moisture. This paper reports on design, fabrication and test of a resonant actuated two-dimensional micro scanning mirror that is hermetically sealed on wafer level. With resonant frequencies of 30kHz and 1kHz, an achievable Theta-D-product of 13mm.deg and low dynamic deformation <20nm RMS it targets Lissajous projection with SVGA-resolution. Inevitable reflexes at the vacuum package surface can be seperated from the projection field by permanent inclination of the micromirror.

Effect of pretreatment with carbon monoxide and ozone on the quality of vacuum packaged beef meats.

PubMed

Lyu, Fei; Shen, Kejing; Ding, Yuting; Ma, Xin

2016-07-01

Beef meats without pretreatment (CK) or pretreated with different volume ratios of carbon monoxide and ozone of 100%CO (T1), 2%O3+98%CO (T2), 5%O3+95%CO (T3) and 10%O3+90%CO (T4) using modified atmosphere packages for 1.5h, after that they were vacuum-packaged and stored in 0°C refrigerator for 46days. The surface color a* values and sensory scores of T1, T2, T3 and T4 were significant higher than CK (p<0.05) during storage. In the mid and later storage, the drip loss, total viable counts (TVC), metmyoglobin (met-Mb), thiobarbituric acid reactive substances (TBARS), total volatile basic nitrogen (TVB-N) and pH of T1, T2, T3 and T4 were significantly lower than CK (p<0.05), and these values of T2, T3 and T4 were significantly lower than T1 in the later storage. In conclusion, O3 in the combination didn't affect the color-developing effect of CO, and could help CO maintain the meat quality. Therefore, the pretreatment of CO combined with O3 at certain concentrations can be a promising technique to maintain the quality of beef meats. Copyright © 2016 Elsevier Ltd. All rights reserved.

Fluid Dynamics of Small, Rugged Vacuum Pumps of Viscous-Drag Type

NASA Technical Reports Server (NTRS)

Russell, John M.

2002-01-01

The need to identify spikes in the concentration of hazardous gases during countdowns to space shuttle launches has led Kennedy Space Center to acquire considerable expertise in the design, construction, and operation of special-purpose gas analyzers of mass-spectrometer type. If such devices could be miniaturized so as to fit in a small airborne package or backpack them their potential applications would include integrated vehicle health monitoring in later-generation space shuttles and in hazardous material detection in airports, to name two examples. The bulkiest components of such devices are vacuum pumps, particularly those that function in the low vacuum range. Now some pumps that operate in the high vacuum range (e.g. molecular-drag and turbomolecular pumps) are already small and rugged. The present work aims to determine whether, on physical grounds, one may or may not adopt the molecular-drag principle to the low-vacuum range (in which case viscous-drag principle is the appropriate term). The deliverable of the present effort is the derivation and justification of some key formulas and calculation methods for the preliminary design of a single-spool, spiral-channel viscous-drag pump.

Characterization and protective efficacy in an animal model of a novel truncated rotavirus VP8 subunit parenteral vaccine candidate.

PubMed

Xue, Miaoge; Yu, Linqi; Che, Yaojian; Lin, Haijun; Zeng, Yuanjun; Fang, Mujin; Li, Tingdong; Ge, Shengxiang; Xia, Ningshao

2015-05-21

The cell-attachment protein VP8* of rotavirus is a potential candidate parenteral vaccine. However, the yield of full-length VP8 protein (VP8*, residues 1-231) expressed in Escherichia coli was low, and a truncated VP8 protein (ΔVP8*, residues 65-231) cannot elicit efficient protective immunity in a mouse model. In this study, tow novel truncated VP8 proteins, VP8-1 (residues 26-231) and VP8-2 (residues 51-231), were expressed in E. coli and evaluated for immunogenicity and protective efficacy, compared with VP8* and ΔVP8*. As well as ΔVP8*, the protein VP8-1 and VP8-2 were successfully expressed in high yield and purified in homogeneous dimeric forms, while the protein VP8* was expressed with lower yield and prone to aggregation and degradation in solution. Although the immunogenicity of the protein VP8*, VP8-1, VP8-2 and ΔVP8* was comparable, immunization of VP8* and VP8-1 elicited significantly higher neutralizing antibody titers than that of VP8-2 and ΔVP8* in mice. Furthermore, when assessed using a mouse maternal antibody model, the efficacy of VP8-1 to protect against rotavirus-induced diarrhea in pups was comparable to that of VP8*, both were dramatically higher than that of VP8-2 and ΔVP8*. Taken together, the novel truncated protein VP8-1, with increased yield, improved homogeneity and high protective efficacy, is a viable candidate for further development of a parenterally administrated prophylactic vaccine against rotavirus infection. Copyright © 2015 Elsevier Ltd. All rights reserved.

Measurement of muscle exudate protein composition as an indicator of beef tenderness

USDA-ARS?s Scientific Manuscript database

The objective was to determine the relationship between the protein composition of muscle exudate and beef tenderness. Strip loins (n = 24) were divided into 3 sections, vacuum packaged, and aged at 4°C. After 0, 7, and 14 days, shear force was measured and muscle exudate was analyzed for protein ...

Nitrite spray treatment to promote red color stability of vacuum packaged beef.

PubMed

Song, Xiao; Cornforth, Daren; Whittier, Dick; Luo, Xin

2015-01-01

Sodium nitrite solutions were sprayed on select grade boneless rib (M. longissimus thoracis) and bottom round (mainly M. biceps femoris) steaks individually, to form bright red nitric oxide myoglobin (NO-Mb) in vacuum packages. Our objective was to determine the optimum level of nitrite in spray for stable raw steak redness, low or no residual nitrite, and low surface pinking (ham-like cured color) after cooking. Results showed that steaks sprayed with 100-350 ppm nitrite solutions had 3.0-3.6g weight gain and a calculated level of 1.3-5.3mg nitrite added/kg steak, but very low (<1 ppm) residual nitrite. Nitrite sprays of 250-350 ppm were optimum for raw steak color during 21 days of storage at 1°C (a*>10; chroma C*>16). Raw steak redness was less stable in round than rib. Visual scores for pinkness after cooking were low, indicating that cooked color at even the highest nitrite treatment (350 ppm) was acceptable. Copyright © 2014 Elsevier Ltd. All rights reserved.

Characterization of wafer-level bonded hermetic packages using optical leak detection

NASA Astrophysics Data System (ADS)

Duan, Ani; Wang, Kaiying; Aasmundtveit, Knut; Hoivik, Nils

2009-07-01

For MEMS devices required to be operated in a hermetic environment, one of the main reliability issues is related to the packaging methods applied. In this paper, an optical method for testing low volume hermetic cavities formed by anodic bonding between glass and SOI (silicon on insulator) wafer is presented. Several different cavity-geometry structures have been designed, fabricated and applied to monitor the hermeticity of wafer level anodic bonding. SOI wafer was used as the cap wafer on which the different-geometry structures were fabricated using standard MEMS technology. The test cavities were bonded using SOI wafers to glass wafers at 400C and 1000mbar pressure inside a vacuum bonding chamber. The bonding voltage varies from 200V to 600V. The bonding strength between glass and SOI wafer was mechanically tested using shear tester. The deformation amplitudes of the cavity cap surface were monitored by using an optical interferometer. The hermeticity of the glass-to-SOI wafer level bonding was characterized through observing the surface deformation in a 6 months period in atmospheric environment. We have observed a relatively stable micro vacuum-cavity.

The effect of carbon monoxide pretreatment exposure time on the colour stability and quality attributes of vacuum packaged beef steaks.

PubMed

Van Rooyen, Lauren Anne; Allen, Paul; Crawley, Sarah M; O'Connor, David I

2017-07-01

The effect of 5% CO pretreatments prior to vacuum packaging of beef striploin steaks (Longissimus thoracis et lumborum, LTL) on quality attributes, primarily colour stability was investigated. The aim was to determine the optimum pretreatment that would induce the desirable red colour, while allowing discoloration to occur by the end of a 28-day display period (2°C), so as to not mask spoilage. A range of pretreatment exposure times (1, 3, 5, 7, 9, 15 and 24h) were applied to steaks using a gas mixture of 5% CO, 60% CO 2 and 35% N 2 . The 5h CO pretreatment exposure time achieved the desirable colour and discoloration reached unacceptable levels (a*=12, C*=16) by the use-by date (28days), thus ensuring consumers' of a reliable visual indication of freshness and addressing concerns about safety. The 5% CO pretreatment had no negative effect on microbiological safety, lipid oxidation, cooking loss and WBSF measurements at the end of storage (P>0.05). Copyright © 2017 Elsevier Ltd. All rights reserved.

Effect of cinnamon essential oil on bacterial diversity and shelf-life in vacuum-packaged common carp (Cyprinus carpio) during refrigerated storage.

PubMed

Zhang, Yuemei; Li, Dongping; Lv, Jian; Li, Qingzheng; Kong, Chunli; Luo, Yongkang

2017-05-16

The present study investigated the effect of cinnamon essential oil on the quality of vacuum-packaged common carp (Cyprinus carpio) fillets stored at 4±1°C in terms of sensory scores, physicochemical characteristics (total volatile basic nitrogen (TVB-N), biogenic amines, and color), and presence of spoilage microbiota. A total of 290,753 bacterial sequences and 162 different genera belonging to 14 phyla were observed by a high-throughput sequencing technique targeting the V3-V4 region of 16S rDNA, which showed a more comprehensive estimate of microbial diversity in carp samples compared with microbial enumeration. Before storage, Macrococcus and Aeromonas were the prevalent populations in the control samples, but cinnamon essential oil decreased the relative abundance of Macrococcus in the treated samples. Variability in the predominant microbiota in different samples during chilled storage was observed. Aeromonas followed by Lactococcus were the major contaminants in the spoiled control samples. Microbial enumeration also observed relatively higher counts of Aeromonas than other spoilage microorganisms. Compared with the control samples, cinnamon essential oil inhibited the growth of Aeromonas and Lactococcus were the predominant components in the treated samples on day 10; plate counts also revealed a relatively high level of lactic acid bacteria during refrigerated storage. However, there were no significant differences (P>0.05) in the composition of dominant microbiota between these two treatments at the end of the shelf-life. Furthermore, cinnamon essential oil treatment was more effective in inhibiting the increase of TVB-N and the accumulation of biogenic amines (especially for putrescine and cadaverine levels). Based primarily on sensory analysis, the use of cinnamon essential oil extended the shelf-life of vacuum-packaged common carp fillets by about 2days. Copyright © 2016 Elsevier B.V. All rights reserved.

Cleavage sites within the poliovirus capsid protein precursors

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

Larsen, G.R.; Anderson, C.W.; Dorner, A.J.

1982-01-01

Partial amino-terminal sequence analysis was performed on radiolabeled poliovirus capsid proteins VP1, VP2, and VP3. A computer-assisted comparison of the amino acid sequences obtained with that predicted by the nucleotide sequence of the poliovirus genome allows assignment of the amino terminus of each capsid protein to a unique position within the virus polyprotein. Sequence analysis of trypsin-digested VP4, which has a blocked amino terminus, demonstrates that VP4 is encoded at or very near to the amino terminus of the polyprotein. The gene order of the capsid proteins is VP4-VP2-VP3-VP1. Cleavage of VP0 to VP4 and VP2 is shown to occurmore » between asparagine and serine, whereas the cleavages that separate VP2/VP3 and VP3/VP1 occur between glutamine and glycine residues. This finding supports the hypothesis that the cleavage of VP0, which occurs during virion morphogenesis, is distinct from the cleavages that separate functional regions of the polyprotein.« less

Meat Processing Plant Microbiome and Contamination Patterns of Cold-Tolerant Bacteria Causing Food Safety and Spoilage Risks in the Manufacture of Vacuum-Packaged Cooked Sausages.

PubMed

Hultman, Jenni; Rahkila, Riitta; Ali, Javeria; Rousu, Juho; Björkroth, K Johanna

2015-10-01

Refrigerated food processing facilities are specific man-made niches likely to harbor cold-tolerant bacteria. To characterize this type of microbiota and study the link between processing plant and product microbiomes, we followed and compared microbiota associated with the raw materials and processing stages of a vacuum-packaged, cooked sausage product affected by a prolonged quality fluctuation with occasional spoilage manifestations during shelf life. A total of 195 samples were subjected to culturing and amplicon sequence analyses. Abundant mesophilic psychrotrophs were detected within the microbiomes throughout the different compartments of the production plant environment. However, each of the main genera of food safety and quality interest, e.g., Leuconostoc, Brochothrix, and Yersinia, had their own characteristic patterns of contamination. Bacteria from the genus Leuconostoc, commonly causing spoilage of cold-stored, modified-atmosphere-packaged foods, were detected in high abundance (up to >98%) in the sausages studied. The same operational taxonomic units (OTUs) were, however, detected in lower abundances in raw meat and emulsion (average relative abundance of 2%±5%), as well as on the processing plant surfaces (<4%). A completely different abundance profile was found for OTUs phylogenetically close to the species Yersinia pseudotuberculosis. These OTUs were detected in high abundance (up to 28%) on the processing plant surfaces but to a lesser extent (<1%) in raw meat, sausage emulsion, and sausages. The fact that Yersinia-like OTUs were found on the surfaces of a high-hygiene packaging compartment raises food safety concerns related to their resilient existence on surfaces. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

The effects of packaging method (vacuum pouch vs. plastic tray) on spoilage in a cook-chill pork-based dish kept under refrigeration.

PubMed

Díaz, Pedro; Garrido, María Dolores; Bañón, Sancho

2010-03-01

The effects of two packaging methods on the spoilage of a cook-chill pork-based dish kept under refrigeration were studied. Raw pork cuts and pre-cooked tomato sauce were packed under vacuum "sous vide" in polyamide-polypropylene pouches (SV) or into translucent polypropylene trays under modified atmosphere (80% N(2)+20% CO(2)) and sealed with a top film (PT). Samples were cooked inside the pack at an oven temperature/time of 70 degrees C/7h, chilled at 3 degrees C and stored at 2 degrees C for up to 90days. Microbial (psychrotrophs, lactic-acid bacteria, Enterobacteriaceae, moulds and yeasts), physical-chemical (pH, water activity and total acidity) and sensory (colour, odour, flavour, texture and acceptance) parameters were determined. Heat penetration was faster in SV (2 degrees C/min) than in PT (1 degrees C/min) (core temperature). Both packaging methods were equally effective in protecting against microbial spoilage for 90 day at 2 degrees C. Minor counts were only detected for lactic-acid bacteria and anaerobic psychrotrophs in SV. No Enterobacteriaceae growth was found. Slight differences between SV and PT in pH and total acidity were observed. SV and PT had similar effects on the sensory preservation of the dishes. A gradual loss of acceptance of the cooked pork and tomato sauce was observed. Rancid flavour in PT and warmed-over-flavour in SV were noted in the final stages of storage. According to acceptance scores, the shelf-life of both SV and PT was 56 days at 2 degrees C. Both packaging methods can be used to manufacture sous vide meat-based dishes subsequently stored under refrigeration for catering use. Copyright 2009 Elsevier Ltd. All rights reserved.

Effects of dietary vitamin E supplementation and packaging on the colour stability of sliced pasteurized beef ham.

PubMed

Houben, J H; van Dijk, A

2001-08-01

The effect of supplementation of vitamin E (2025 IU animal(-1) day(-1)) in the diet of beef bulls on the colour stability of pasteurized beef ham was studied. Control and enriched diets were provided for the last 136 days before slaughter. Pasteurized hams were manufactured from Mm. semitendinosus from eight animals per dietary group. Half of the samples of sliced ham from control (CON) and supplemented (SUP) bulls were packaged under vacuum (VAC) and half in low-oxygen modified atmosphere packs (FOG, gas mixture: CO(2)/N(2)=50/50). The packages were kept under constant illumination for 28 days at 8°C. During storage, the number of colony-forming units (CFU) reached a maximum of 5x10(7) g(-1). The microflora was dominated by lactic acid bacteria. The supplementation with vitamin E showed no effect on microbial growth. Lipid oxidation was stable during storage. A significant difference between both dietary groups was detected for the decrease in the redness values during storage. Redness values of CON vacuum-packaged samples decreased (P < 0.01) with time, whereas those for the SUP products only tended to decrease. The redness values of FOG-packed ham were higher than those of VAC-packed ham at the end of the display period, irrespective of the dietary group. Overall, colour appeared to be more stable in the FOG-packed products than in the VAC-packed products. It can be concluded that dietary supplementation of bulls with vitamin E appears to offer only a minor improvement in colour stability over current feeding regimens when the Mm. semitendinosus are used to make cured, pasteurized ham-type products.

Design, assembly, and testing of a high-resolution relay lens used for holography with operation at both doubled and tripled Nd:YAG laser wavelengths

NASA Astrophysics Data System (ADS)

Malone, Robert M.; Capelle, Gene A.; Cox, Brian C.; Frogget, Brent C.; Grover, Mike; Kaufman, Morris I.; Pazuchanics, Peter; Sorenson, Danny S.; Stevens, Gerald D.; Tibbitts, Aric; Turley, William D.

2009-08-01

The design and assembly of a nine-element lens that achieves >2000 lp/mm resolution at a 355-nm wavelength (ultraviolet) has been completed. By adding a doublet to this lens system, operation at a 532-nm wavelength (green) with >1100 lp/mm resolution is achieved. This lens is used with high-power laser light to record holograms of fast-moving ejecta particles from a shocked metal surface located inside a test package. Part of the lens and the entire test package are under vacuum with a 1-cm air gap separation. Holograms have been recorded with both doubled and tripled Nd:YAG laser light. The UV operation is very sensitive to the package window's tilt. If this window is tilted by more than 0.1 degrees, the green operation performs with better resolution than that of the UV operation. The setup and alignment are performed with green light, but the dynamic recording can be done with either UV light or green light. A resolution plate can be temporarily placed inside the test package so that a television microscope located beyond the hologram position can archive images of resolution patterns that prove that the calibration wires, interference filter, holographic plate, and relay lenses are in their correct positions. Part of this lens is under vacuum, at the point where the laser illumination passes through a focus. Alignment and tolerancing of this high-resolution lens are presented. Resolution variation across the 12-mm field of view and throughout the 5-mm depth of field is discussed for both wavelengths.

Improving Shipboard Applications of Non-Intrusive Load Monitoring

DTIC Science & Technology

2008-06-01

EVENTS_LIST (EventNum) .EVENTCODE) case 0 %Unclassifiable case 1 %VP On STATE.VP.ON = STATE.VP.ON + 1; STATE.VP.START = true; PICTURES.RUNNING.VP.SHOW...true; case 2 %VP Off STATE.VP.STOP = true; PICTURES. RUNNING.VP. SHOW false; case 3 %DP On 149 STATE.DP.ON = STATE.DP.ON + 1; STATE.DP.START = true...PICTURES.RUNNING.DP.SHOW true; case 4 %DP Off STATE.DP.STOP = true; PICTURES.RUNNING.DP.SHOW = false; case 5 %VP On (Clogged) STATE.VP.ON = STATE.VP.ON

Molecular analysis of partial VP-2 gene amplified from rectal swab samples of diarrheic dogs in Pakistan confirms the circulation of canine parvovirus genetic variant CPV-2a and detects sequences of feline panleukopenia virus (FPV).

PubMed

Ahmed, Nisar; Riaz, Adeel; Zubair, Zahra; Saqib, Muhammad; Ijaz, Sehrish; Nawaz-Ul-Rehman, Muhammad Shah; Al-Qahtani, Ahmed; Mubin, Muhammad

2018-03-15

The infection in dogs due to canine parvovirus (CPV), is a highly contagious one with high mortality rate. The present study was undertaken for a detailed genetic analysis of partial VP2 gene i.e., 630 bp isolated from rectal swab samples of infected domestic and stray dogs from all areas of district Faisalabad. Monitoring of viruses is important, as continuous prevalence of viral infection might be associated with emergence of new virulent strains. In the present study, 40 rectal swab samples were collected from diarrheic dogs from different areas of district Faisalabad, Pakistan, in 2014-15 and screened for the presence of CPV by immunochromatography. Most of these dogs were stray dogs showing symptoms of diarrhea. Viral DNA was isolated and partial VP2 gene was amplified using gene specific primer pair Hfor/Hrev through PCR. Amplified fragments were cloned in pTZ57R/T (Fermentas) and completely sequenced. Sequences were analyzed and assembled by the Lasergene DNA analysis package (v8; DNAStar Inc., Madison, WI, USA). The results with immunochromatography showed that 33/40 (82%) of dogs were positive for CPV. We were able to amplify a fragment of 630 bp from 25 samples. In 25 samples the sequences of CPV-2a were detected showing the amino acid substitution Ser297Ala and presence of amino acid (426-Asn) in partial VP2 protein. Interestingly the BLAST analysis showed the of feline panleukopenia virus (FPV) sequences in 3 samples which were already positive for new CPV-2a, with 99% sequence homology to other FPV sequences present in GenBank. Phylogenetic analysis showed clustering of partial CPV-VP-2 gene with viruses from China, India, Japan and Uruguay identifying a new variant, whereas the 3 FPV sequences showed immediate ancestral relationship with viruses from Portugal, South Africa and USA. Interesting observation was that CPV are clustering away from the commercial vaccine strains. In this work we provide a better understanding of CPV prevailing in Pakistan at molecular level. The detection of FPV could be a case of real co-infection or a case of dual presence, due to ingestion of contaminated food.

Packaging of MEMS/MOEMS and nanodevices: reliability, testing, and characterization aspects

NASA Astrophysics Data System (ADS)

Tekin, Tolga; Ngo, Ha-Duong; Wittler, Olaf; Bouhlal, Bouchaib; Lang, Klaus-Dieter

2011-02-01

The last decade witnessed an explosive growth in research and development efforts devoted to MEMS devices and packaging. The successfully developed MEMS devices are, for example inkjet, pressure sensors, silicon microphones, accelerometers, gyroscopes, MOEMS, micro fuel cells and emerging MEMS. For the next decade, MEMS/MOEMS and nanodevice based products will penetrate into IT, telecommunications, automotive, defense, life sciences, medical and implantable applications. Forecasts say the MEMS market to be $14 billion by 2012. The packaging cost of MEMS/MOEMS products in general is about 70 percent. Unlike today's electronics IC packaging, their packaging are custom-built and difficult due to the moving structural elements. In order for the moving elements of a MEMS device to move effectively in a well-controlled atmosphere, hermetic sealing of the MEMS device in a cap is necessary. For some MEMS devices, such as resonators and gyroscopes, vacuum packaging is required. Usually, the cap is processed at the wafer level, and thus MEMS packaging is truly a wafer level packaging. In terms of MEMS/MOEMS and nanodevice packaging, there are still many critical issues need to be addressed due to the increasing integration density supported by 3D heterogeneous integration of multi-physic components/layers consisting of photonics, electronics, rf, plasmonics, and wireless. The infrastructure of MEMS/MOEMS and nanodevices and their packaging is not well established yet. Generic packaging platform technologies are not available. Some of critical issues have been studied intensively in the last years. In this paper we will discuss about processes, reliability, testing and characterization of MEMS/MOEMS and nanodevice packaging.

Expression and characterization of highly antigenic domains of chicken anemia virus viral VP2 and VP3 subunit proteins in a recombinant E. coli for sero-diagnostic applications

PubMed Central

2013-01-01

Background Chicken anemia virus (CAV) is an important viral pathogen that causes anemia and severe immunodeficiency syndrome in chickens worldwide. Generally, CAV infection occurs via vertical transmission in young chicks that are less than two weeks old, which are very susceptible to the disease. Therefore, epidemiological investigations of CAV infection and/or the evaluation of the immunization status of chickens is necessary for disease control. Up to the present, systematically assessing viral protein antigenicity and/or determining the immunorelevant domain(s) of viral proteins during serological testing for CAV infection has never been performed. The expression, production and antigenic characterization of CAV viral proteins such as VP1, VP2 and VP3, and their use in the development of diagnostic kit would be useful for CAV infection prevention. Results Three CAV viral proteins VP1, VP2 and VP3 was separately cloned and expressed in recombinant E. coli. The purified recombinant CAV VP1, VP2 and VP3 proteins were then used as antigens in order to evaluate their reactivity against chicken sera using indirect ELISA. The results indicated that VP2 and VP3 show good immunoreactivity with CAV-positive chicken sera, whereas VP1 was found to show less immunoreactivity than VP2 and VP3. To carry out the further antigenic characterization of the immunorelevant domains of the VP2 and VP3 proteins, five recombinant VP2 subunit proteins (VP2-435N, VP2-396N, VP2-345N, VP2-171C and VP2-318C) and three recombinant VP3 subunit proteins (VP3-123N, VP3-246M, VP3-366C), spanning the defined regions of VP2 and VP3 were separately produced by an E. coli expression system. These peptides were then used as antigens in indirect ELISAs against chicken sera. The results of these ELISAs using truncated recombinant VP2 and VP3 subunit proteins as coating antigen showed that VP2-345N, VP2-396N and VP3-246M gave good immunoreactivity with CAV-positive chicken sera compared to the other subunit proteins. Moreover, the VP2-396N and VP2-345 based ELISAs had better sensitivity (97.5%) and excellent specificity (100%) during serodiagnosis testing using a mean plus three standard deviations cut-off. The VP3-246M based ELISA showed a sensitivity of 85% and a specificity of 100% at the same cut-off value. Conclusions This is the first report to systematically assess the antigenic characteristics of CAV viral proteins for sero-diagnosis purposes. Purified recombinant VP2-396N and VP2-345N subunit proteins, which span defined regions of VP2, were demonstrated to have good antigenicity and higher sensitivities than VP3-246M and were able to recognize CAV-positive chicken serum using an ELISA assay. The defined antigenicity potential of these chimeric subunit proteins produced by expression in E. coli seem to have potential and could be useful in the future for the development of the CAV diagnostic tests based on a subunit protein ELISA system. PMID:23937712

VP24-Karyopherin Alpha Binding Affinities Differ between Ebolavirus Species, Influencing Interferon Inhibition and VP24 Stability

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

Schwarz, Toni M.; Edwards, Megan R.; Diederichs, Audrey

ABSTRACT Zaire ebolavirus(EBOV),Bundibugyo ebolavirus(BDBV), andReston ebolavirus(RESTV) belong to the same genus but exhibit different virulence properties. VP24 protein, a structural protein present in all family members, blocks interferon (IFN) signaling and likely contributes to virulence. Inhibition of IFN signaling by EBOV VP24 (eVP24) involves its interaction with the NPI-1 subfamily of karyopherin alpha (KPNA) nuclear transporters. Here, we evaluated eVP24, BDBV VP24 (bVP24), and RESTV VP24 (rVP24) interactions with three NPI-1 subfamily KPNAs (KPNA1, KPNA5, and KPNA6). Using purified proteins, we demonstrated that each VP24 binds to each of the three NPI-1 KPNAs. bVP24, however, exhibited approximately 10-fold-lower KPNA bindingmore » affinity than either eVP24 or rVP24. Cell-based assays also indicate that bVP24 exhibits decreased KPNA interaction, decreased suppression of IFN induced gene expression, and a decreased half-life in transfected cells compared to eVP24 or rVP24. Amino acid sequence alignments between bVP24 and eVP24 also identified residues within and surrounding the previously defined eVP24-KPNA5 binding interface that decrease eVP24-KPNA affinity or bVP24-KPNA affinity. VP24 mutations that lead to reduced KPNA binding affinity also decrease IFN inhibition and shorten VP24 half-lives. These data identify novel functional differences in VP24-KPNA interaction and reveal a novel impact of the VP24-KPNA interaction on VP24 stability. IMPORTANCEThe interaction of Ebola virus (EBOV) VP24 protein with host karyopherin alpha (KPNA) proteins blocks type I interferon (IFN) signaling, which is a central component of the host innate immune response to viral infection. Here, we quantitatively compared the interactions of VP24 proteins from EBOV and two members of theEbolavirusgenus, Bundibugyo virus (BDBV) and Reston virus (RESTV). The data reveal lower binding affinity of the BDBV VP24 (bVP24) for KPNAs and demonstrate that the interaction with KPNA modulates inhibition of IFN signaling and VP24 stability. The effect of KPNA interaction on VP24 stability is a novel functional consequence of this virus-host interaction, and the differences identified between viral species may contribute to differences in pathogenesis.« less

VP24-Karyopherin Alpha Binding Affinities Differ between Ebolavirus Species, Influencing Interferon Inhibition and VP24 Stability.

PubMed

Schwarz, Toni M; Edwards, Megan R; Diederichs, Audrey; Alinger, Joshua B; Leung, Daisy W; Amarasinghe, Gaya K; Basler, Christopher F

2017-02-15

Zaire ebolavirus (EBOV), Bundibugyo ebolavirus (BDBV), and Reston ebolavirus (RESTV) belong to the same genus but exhibit different virulence properties. VP24 protein, a structural protein present in all family members, blocks interferon (IFN) signaling and likely contributes to virulence. Inhibition of IFN signaling by EBOV VP24 (eVP24) involves its interaction with the NPI-1 subfamily of karyopherin alpha (KPNA) nuclear transporters. Here, we evaluated eVP24, BDBV VP24 (bVP24), and RESTV VP24 (rVP24) interactions with three NPI-1 subfamily KPNAs (KPNA1, KPNA5, and KPNA6). Using purified proteins, we demonstrated that each VP24 binds to each of the three NPI-1 KPNAs. bVP24, however, exhibited approximately 10-fold-lower KPNA binding affinity than either eVP24 or rVP24. Cell-based assays also indicate that bVP24 exhibits decreased KPNA interaction, decreased suppression of IFN induced gene expression, and a decreased half-life in transfected cells compared to eVP24 or rVP24. Amino acid sequence alignments between bVP24 and eVP24 also identified residues within and surrounding the previously defined eVP24-KPNA5 binding interface that decrease eVP24-KPNA affinity or bVP24-KPNA affinity. VP24 mutations that lead to reduced KPNA binding affinity also decrease IFN inhibition and shorten VP24 half-lives. These data identify novel functional differences in VP24-KPNA interaction and reveal a novel impact of the VP24-KPNA interaction on VP24 stability. The interaction of Ebola virus (EBOV) VP24 protein with host karyopherin alpha (KPNA) proteins blocks type I interferon (IFN) signaling, which is a central component of the host innate immune response to viral infection. Here, we quantitatively compared the interactions of VP24 proteins from EBOV and two members of the Ebolavirus genus, Bundibugyo virus (BDBV) and Reston virus (RESTV). The data reveal lower binding affinity of the BDBV VP24 (bVP24) for KPNAs and demonstrate that the interaction with KPNA modulates inhibition of IFN signaling and VP24 stability. The effect of KPNA interaction on VP24 stability is a novel functional consequence of this virus-host interaction, and the differences identified between viral species may contribute to differences in pathogenesis. Copyright © 2017 American Society for Microbiology.

Influence of Barley Sourdough and Vacuum Cooling on Shelf Life Quality of Partially Baked Bread

PubMed Central

2017-01-01

Summary Driven by the bakery industry urge to satisfy consumer demand for fresh, diverse and high quality bakery products, we investigated the influence of barley sourdough and vacuum cooling on shelf life quality of partially baked bread stored in modified atmosphere packaging at ambient conditions. Barley sourdough was fermented with Lactobacillus reuteri (DSM 20016, F275). Partially baked bread with sourdough was microbiologically acceptable during 30 days of storage, while bread without sourdough had detectable mould on the 30th day. Stored bread samples were rebaked after 1, 8, 15, 22 and 30 days to determine moisture content, physical and sensorial properties. Moisture loss (5%) was detected on the 15th day, after which it remained stable until the end of investigated storage period. Nevertheless, textural quality of stored bread continuously declined due to crumb firming. Bread flavour did not change during mould-free storage time. The principal component analysis identified major differences in the flavour of sour and control bread, also in crumb firmness and moisture content of samples. This study indicates the positive role of barley sourdough fermented with L. reuteri in improving crumb texture for at least 15 days, and ensuring mould- and bacteria-free partially baked bread for 30 days. Vacuum cooling combined with sourdough improved bread shape, porosity, and reduced sour taste, crust colouring and crumbliness. Hence, it can successfully extend shelf life quality of partially baked bread in modified atmosphere packaging. PMID:29540981

Enhancement of VP6 immunogenicity and protective efficacy against rotavirus by VP2 in a genetic immunization.

PubMed

Lopez-Guerrero, D V; Arias, N; Gutierrez-Xicotencatl, L; Chihu-Amparan, L; González, A; Pedroza-Saavedra, A; Rosas-Salgado, G; Villegas-Garcia, J C; Badillo-Godinez, O; Fernandez, G; Lopez, S; Esquivel-Guadarrama, F

2018-05-24

VP2/VP6 virus like particles (VLPs) are very effective in inducing protection against the rotavirus infection in animal models. Individually, VP6 can also induce protection. However, there is no information about the immunogenicity of VP2. The aim of this work was to evaluate the efficacy of DNA vaccines codifying for VP2 or VP6, alone or combined, to induce protection against the rotavirus infection. Murine rotavirus VP2 and VP6 genes were cloned into the pcDNA3 vector. Adult BALB/c mice were inoculated three times by intramuscular (i.m.) injections with 100 or 200µg of pcDNA3-VP2 or pcDNA3-VP6 alone or co-administered with 100µg of pcDNA3-VP2/100µg of pcDNA3-VP6. Two weeks after the last inoculation, mice were challenged with the wild type murine rotavirus strain epizootic diarrhea of infant mice (EDIM wt ). We found that both plasmids, pcDNA3-VP2 and pcDNA3-VP6, were able to induce rotavirus-specific serum antibodies, but not intestinal rotavirus-specific IgA; only 200µg of pcDNA3-VP6 induced 35% protection against the infection. A similar level of protection was found when mice were co-administered with 100µg of pcDNA3-VP2/100µg of pcDNA3-VP6 (1:1 ratio). However, the best protection (up to 58%) occurred when mice were inoculated with 10µg of pcDNA3-VP2/100µg of pcDNA3-VP6 (1:10 ratio). These results indicate that the DNA plasmid expressing VP6 is a better vaccine candidate that the one expressing VP2. However, when co-expressed, VP2 potentiates the immunogenicity and protective efficacy of VP6. Copyright © 2017. Published by Elsevier Ltd.

Cryo-EM maps reveal five-fold channel structures and their modification by gatekeeper mutations in the parvovirus minute virus of mice (MVM) capsid.

PubMed

Subramanian, Suriyasri; Organtini, Lindsey J; Grossman, Alec; Domeier, Phillip P; Cifuente, Javier O; Makhov, Alexander M; Conway, James F; D'Abramo, Anthony; Cotmore, Susan F; Tattersall, Peter; Hafenstein, Susan

2017-10-01

In minute virus of mice (MVM) capsids, icosahedral five-fold channels serve as portals mediating genome packaging, genome release, and the phased extrusion of viral peptides. Previous studies suggest that residues L172 and V40 are essential for channel function. The structures of MVMi wildtype, and mutant L172T and V40A virus-like particles (VLPs) were solved from cryo-EM data. Two constriction points, termed the mid-gate and inner-gate, were observed in the channels of wildtype particles, involving residues L172 and V40 respectively. While the mid-gate of V40A VLPs appeared normal, in L172T adjacent channel walls were altered, and in both mutants there was major disruption of the inner-gate, demonstrating that direct L172:V40 bonding is essential for its structural integrity. In wildtype particles, residues from the N-termini of VP2 map into claw-like densities positioned below the channel opening, which become disordered in the mutants, implicating both L172 and V40 in the organization of VP2 N-termini. Copyright © 2017 Elsevier Inc. All rights reserved.

Thematic mapper flight model preshipment review data package. Volume 3, part A: System data

NASA Technical Reports Server (NTRS)

1982-01-01

Results of vibration, acoustical noise, and thermal vacuum are described as well as tests studies of EMI/EMC and mass properties conducted for thematic mapper systems integration. Liens are summarized and the engineering change proposal status is presented. Requests for deviation/waiver are included along with failure and nonforming material reports.

Innovative on-chip packaging applied to uncooled IRFPA

NASA Astrophysics Data System (ADS)

Dumont, Geoffroy; Arnaud, Agnès; Impérinetti, Pierre; Vialle, Claire; Rabaud, Wilfried; Goudon, Valérie; Yon, Jean-Jacques

2008-04-01

The Laboratoire Infrarouge (LIR) of the Laboratoire d'Electronique et de Technologie de l'Information (LETI) has been involved in the development of microbolometers for over fifteen years. Two generations of technology have been transferred to ULIS and LETI is still working to improve performances of low cost detectors. Simultaneously, packaging still represents a significant part of detectors price. Reducing production costs would contribute to keep on extending applications of uncooled IRFPA to high volume markets like automotive. Therefore LETI is developing an on-chip packaging technology dedicated to microbolometers. This paper presents an original microcap structure that enables the use of IR window materials as sealing layers to maintain the expected vacuum level. The modelling and integration of an IR window suitable for this structure is also presented. This monolithic packaging technology is performed in a standard collective way, in continuation of bolometers' technology. The CEA-LETI, MINATEC presents status of these developments concerning this innovating technology including optical simulations results and SEM views of technical realizations.

Stability of lamb loin stored under refrigeration and packed in different modified atmosphere packaging systems.

PubMed

Fernandes, Rafaella de Paula Paseto; Freire, Maria Teresa de Alvarenga; de Paula, Elisa Silva Maluf; Kanashiro, Ana Livea Sayuri; Catunda, Fernanda Antunes Pinto; Rosa, Alessandra Fernandes; Balieiro, Júlio Cesar de Carvalho; Trindade, Marco Antonio

2014-01-01

The aim of the present study was to evaluate the effect of different modified atmosphere packaging (MAP) systems (vacuum, 75% O2+25% CO2 and 100% CO2) on the stability of lamb loins stored at 1±1°C for 28 days. Microbiological (counts of aerobic and anaerobic psychrotrophic microorganisms, coliform at 45°C, coagulase-positive staphylococci and lactic acid bacteria and presence of Salmonella), physical and chemical (thiobarbituric acid reactive substances [TBARS], objective color, pH, water loss from cooking [WLC] and shear force), sensory (acceptance testing using a 9-point hedonic scale) and gas composition analyses were performed. Lamb meat remained stable with respect to the majority of the evaluated physical and chemical indexes and within the standards established by Brazilian legislation for pathogenic microorganisms throughout the storage period in all three packaging systems. However, with respect to psychrotrophic microorganisms, 100% CO2 packaging system provided increased stability despite presenting lower appearance preference. © 2013.

Influence of smoking and packaging methods on lipid stability and microbial quality of Capelin (Mallotus villosus) and Sardine (Sardinella gibossa)

PubMed Central

Cyprian, Odoli O; Van Nguyen, Minh; Sveinsdottir, Kolbrun; Jonsson, Asbjorn; Tomasson, Tumi; Thorkelsson, Gudjon; Arason, Sigurjon

2015-01-01

Lipid and microbial quality of smoked capelin (two groups differing in lipid content) and sardine was studied, with the aim of introducing capelin in the smoked sardine markets. Lipid hydrolysis (phospholipid and free fatty acids) and oxidation index (hydroperoxides and thiobarbituric acid-reactive substances), fatty acid composition, and total viable count were measured in raw and packaged smoked fish during chilled storage (day 2, 10, 16, 22, 28). Lipid hydrolysis was more pronounced in low lipid capelin, whereas accelerated lipid oxidation occurred in high lipid capelin. Muscle lipid was less stable in sardine than capelin. Essential polyunsaturated fatty acids (eicosapentaenoic acid and docosahexaenoic acid) constituted 12% of fatty acids in capelin and 19% in sardine. Vacuum packaging as well as hot smoking retarded bacterial growth, recording counts of ≤log 5 CFU/g compared to ≥log 7CFU/g in cold smoked air packaged. Smoked low lipid capelin was considered an alternative for introduction in smoked sardine markets. PMID:26405526

Effect of the dietary supplementation with Vitamin E on colour stability of packaged, sliced pasteurized ham.

PubMed

Houben, J H; Gerris, C V

1998-12-01

The effect of dietary supplementation with vitamin E (200IU kg(-1) feed) on the colour stability of pasteurized ham was studied. Pigs were fed on control and enriched diets for the last 12 weeks before slaughter. Pasteurized ham was manufactured from the hams from 6 barrows and 6 gilts per dietary group. Half of the samples of sliced ham from control and supplemented pigs were packaged under vacuum (VAC) and half in low-oxygen modified atmosphere packs (FOG, gas mixture: CO(2)/N(2)=60/40). Half the packages were kept under constant illumination and the other half in the dark, both for 22 days at 7°C. The redness component of the VAC-packaged ham prepared from vitamin E-supplemented pigs was slightly more stable than that of comparably packaged ham prepared from control pigs. The opposite was observed for the FOG-packaged products. Overall, colour changes were greater in the ham in FOG-packs than in the ham in VAC-packs. In addition, the colour of the FOG-packaged ham was clearly affected by illumination, whereas the colour of the VAC-packaged ham appeared more stable. It is concluded that dietary supplementation of pigs with vitamin E does not appear to offer significant advantages over currently used feeding regimens with regard to the quality of the ham produced.

Microbial profiles of commercial, vacuum-packaged, fresh pork of normal or short storage life.

PubMed

Holley, Richard A; Peirson, Michael D; Lam, Jocelyn; Tan, Kit Bee

2004-12-01

The microbial ecology of fresh vacuum-packed pork cuts during storage at -1.5 degrees C for up to 45 days was examined to characterize rates of microbial growth and pH changes in commercially prepared products of normal storage quality. Pork loins in commercial distribution with odour defects were also studied to determine a possible cause of the defects and avoid future problems. In addition, microbial profiles of pork cuts from two plants were compared, after storage for 25 days at -1.5 degrees C, to identify possible reasons for differences in the storage life of product from the plants. The effects of a change in sanitation procedures on the microbial populations of products stored for 25 days were also studied. With normal product, microbial growth in different packages progressed at different rates, reflecting differences in initial levels of bacterial contamination. All samples in the study reached 8 weeks without apparent organoleptic change and samples carried 5.8+/-1.2 log bacteria cm(-2) (mean+/-S.D.). The flora of loins with the odour defect were predominately lactic acid bacteria (LAB) and carnobacteria, but they contained large fractions of Enterobacteriaceae <35 days after packaging. Aeromonas spp. and Shewanella spp. were likely responsible for the sulfide-putrid smell of these spoiled products, but species of Enterobacteriaceae and lactic acid bacteria could have contributed to spoilage. Comparison of microbial groups present in 16 other cuts, half from each of two commercial plants, which were stored for 25 days at -1.5 degrees C, showed that larger fractions of Enterobacteriaceae were present in samples from the plant having difficulty achieving the desired storage life. Additional bacterial samples from 12 cuts supplied by the latter plant obtained after adoption of an acid sanitizer step in the plant cleaning regimen, and also stored for 25 days at -1.5 degrees C, yielded few Enterobacteriaceae, Aeromonas or Shewanella. Use of an acid sanitizer in plant cleaning may be a means of controlling alkali-tolerant bacteria such as Aeromonas or Shewanella which can contaminate pork cuts and spoil vacuum-packaged product. The fraction of Enterobacteriaceae in bacteria populations on fresh pork stored for 25 days at -1.5 degrees C may be a useful indicator of the effectiveness of plant sanitation.

Antimicrobial evaluation of novel poly-lactic acid based nanocomposites incorporated with bioactive compounds in-vitro and in refrigerated vacuum-packed cooked sausages.

PubMed

Rezaeigolestani, Mohammadreza; Misaghi, Ali; Khanjari, Ali; Basti, Afshin Akhondzadeh; Abdulkhani, Ali; Fayazfar, Samira

2017-11-02

Biodegradability and antimicrobial activity of food packaging materials are among the most attractive parameters in modern food industries. In order to develop biodegradable poly-lactic acid (PLA) film to antibacterial nanocomposites, different concentration of Zataria multiflora Bioss. essential oil (ZME), propolis ethanolic extract (PEE) and cellulose nanofiber (CNF) were incorporated to the polymer by solvent casting method. The resulting films were characterized by mechanical and physical tests and their antimicrobial application was evaluated in-vitro against four common foodborne pathogens and in vacuum-packed cooked sausages during refrigerated storage. Mechanical examination revealed that addition of ZME and PEE made films more flexible and incorporation of CNF improved almost all mechanical parameters tested. Moreover, according to physical analysis, incorporation of 0.5% v/v ZME to the composite primary solutions improved water vapor permeability of the resulting films. Almost all of the active films were effective against the tested bacteria except for PLA/PEE films, and maximum antibacterial effects recorded for the films containing both ZME and PEE. Based on the microbiological and sensory evaluation of the sausages, all of the PLA/1%ZME/PEE composites increased the shelf life to >40days. The results indicate that incorporation of natural antimicrobial substances such as ZME and PEE to packaging material could be an interesting approach in development of active packaging material without significant negative effect on polymer technical properties. Copyright © 2017 Elsevier B.V. All rights reserved.

Microbial growth, communities and sensory characteristics of vacuum and modified atmosphere packaged lamb shoulders.

PubMed

Kiermeier, Andreas; Tamplin, Mark; May, Damian; Holds, Geoff; Williams, Michelle; Dann, Alison

2013-12-01

Packaging fresh lamb in a vacuum (VAC) versus a 100% CO2 modified atmosphere (MAP) may influence product shelf-life and the bacterial communities. While VAC is a common packing method and 100% CO2 MAP is used in some countries, there is little information about how these different techniques affect the growth of spoilage bacteria and sensory attributes of lamb. The aim of this study was to assess changes in microbiological and organoleptic properties, and determine differences in microbial communities by terminal restriction fragment length polymorphism (TRFLP) and 454 pyrosequencing, in bone-in (BI) and bone-out (BO) MAP- and VAC-packed lamb shoulders stored at -0.3 °C over 12 wk. VAC and MAP lamb shoulders were acceptable in sensory test scores over 12 wk of storage at -0.3 °C, despite total viable count (TVC) and lactic acid bacteria (LAB) levels increasing to 8 log10 CFU/cm(2) for VAC lamb and 4-6 log10 CFU/cm(2) for MAP lamb. Similar to the sensory results, there were no significant differences in microbial communities between BI and BO product. However, types of bacteria were different between VAC and MAP packaging. Specifically, while VAC shoulder became dominated by Carnobacterium spp. in the middle of the storage period, the MAP shoulder microbial population remained similar from the start until later storage times. Copyright © 2013 Elsevier Ltd. All rights reserved.

The study of the intercellular trafficking of the fusion proteins of herpes simplex virus protein VP22.

PubMed

Xue, Xiaodong; Huang, Jianhua; Wang, Huishan

2014-01-01

Genetic modifications can improve the therapeutic efficacy of mesenchymal stem cell (MSC) transplantation in myocardial infarction. However, so far, the efficiency of MSC modification is very low. Seeking for a more efficient way of MSC modification, we investigated the possibility of employing the intercellular trafficking capacity of the herpes simplex virus type-1 tegument protein VP22 on the enhancement of MSC modification. Plasmids pVP22-myc, pVP22-EGFP, pEGFP-VP22, pVP22-hBcl-xL and phBcl-xL-VP22 were constructed for the expressions of the myc-tagged VP22 and the fusion proteins VP22-EGFP, EGFP-VP22, VP22-hBcl-xL and hBcl-xL-VP22. MSCs were isolated from rat bone marrow and the surface markers were identified by Flowcytometry. COS-1 cells were transfected with the above plasmids and co-cultured with untransfected MSCs, the intercellular transportations of the constructed proteins were studied by immunofluorescence. The solubility of VP22-hBcl-xL and hBcl-xL-VP22 was analyzed by Western blot. VP22-myc could be expressed in and spread between COS-1 cells, which indicates the validity of our VP22 expression construct. Flowcytometry analysis revealed that the isolated MSCs were CD29, CD44, and CD90 positive and were negative for the hematopoietic markers, CD34 and CD45. The co-culturing and immunofluorescence assay showed that VP22-myc, VP22-EGFP and EGFP-VP22 could traffic between COS-1 cells and MSCs, while the evidence of intercellular transportation of VP22-hBcl-xL and hBcl-xL-VP22 was not detected. Western blot analysis showed that VP22-hBcl-xL and hBcl-xL-VP22 were both insoluble in the cell lysate suggesting interactions of the fusion proteins with other cellular components. The intercellular trafficking of VP22-myc, VP22-EGFP and EGFP-VP22 between COS-1 cells and MSCs presents an intriguing prospect in the therapeutic application of VP22 as a delivery vehicle which enhances genetic modifications of MSCs. However, VP22-hBcl-xL and hBcl-xL-VP22 failed to spread between cells, which are due to the insolubility of the fusion protein incurred by interactions with other cellular components.

The Study of the Intercellular Trafficking of the Fusion Proteins of Herpes Simplex Virus Protein VP22

PubMed Central

Xue, Xiaodong; Huang, Jianhua; Wang, Huishan

2014-01-01

Background Genetic modifications can improve the therapeutic efficacy of mesenchymal stem cell (MSC) transplantation in myocardial infarction. However, so far, the efficiency of MSC modification is very low. Seeking for a more efficient way of MSC modification, we investigated the possibility of employing the intercellular trafficking capacity of the herpes simplex virus type-1 tegument protein VP22 on the enhancement of MSC modification. Methods Plasmids pVP22-myc, pVP22-EGFP, pEGFP-VP22, pVP22-hBcl-xL and phBcl-xL-VP22 were constructed for the expressions of the myc-tagged VP22 and the fusion proteins VP22-EGFP, EGFP-VP22, VP22-hBcl-xL and hBcl-xL-VP22. MSCs were isolated from rat bone marrow and the surface markers were identified by Flowcytometry. COS-1 cells were transfected with the above plasmids and co-cultured with untransfected MSCs, the intercellular transportations of the constructed proteins were studied by immunofluorescence. The solubility of VP22-hBcl-xL and hBcl-xL-VP22 was analyzed by Western blot. Results VP22-myc could be expressed in and spread between COS-1 cells, which indicates the validity of our VP22 expression construct. Flowcytometry analysis revealed that the isolated MSCs were CD29, CD44, and CD90 positive and were negative for the hematopoietic markers, CD34 and CD45. The co-culturing and immunofluorescence assay showed that VP22-myc, VP22-EGFP and EGFP-VP22 could traffic between COS-1 cells and MSCs, while the evidence of intercellular transportation of VP22-hBcl-xL and hBcl-xL-VP22 was not detected. Western blot analysis showed that VP22-hBcl-xL and hBcl-xL-VP22 were both insoluble in the cell lysate suggesting interactions of the fusion proteins with other cellular components. Conclusions The intercellular trafficking of VP22-myc, VP22-EGFP and EGFP-VP22 between COS-1 cells and MSCs presents an intriguing prospect in the therapeutic application of VP22 as a delivery vehicle which enhances genetic modifications of MSCs. However, VP22-hBcl-xL and hBcl-xL-VP22 failed to spread between cells, which are due to the insolubility of the fusion protein incurred by interactions with other cellular components. PMID:24955582

A digital miniature x-ray tube with a high-density triode carbon nanotube field emitter

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

Jeong, Jin-Woo; Kang, Jun-Tae; Choi, Sungyoul

2013-01-14

We have fabricated a digital miniature x-ray tube (6 mm in diameter and 32 mm in length) with a high-density triode carbon nanotube (CNT) field emitter for special x-ray applications. The triode CNT emitter was densely formed within a diameter of below 4 mm with the focusing-functional gate. The brazing process enables us to obtain and maintain a desired vacuum level for the reliable electron emission from the CNT emitters after the vacuum packaging. The miniature x-ray tube exhibited a stable and reliable operation over 250 h in a pulse mode at an anode voltage of above 25 kV.

Luminescence of BaBrI and SrBrI single crystals doped with Eu2+

NASA Astrophysics Data System (ADS)

Shalaev, A. A.; Shendrik, R.; Myasnikova, A. S.; Bogdanov, A.; Rusakov, A.; Vasilkovskyi, A.

2018-05-01

The crystal growth procedure and luminescence properties of pure and Eu2+-doped BaBrI and SrBrI crystals are reported. Emission and excitation spectra were recorded under ultraviolet and vacuum ultraviolet excitations. The energy of the first Eu2+ 4f-5d transition and SrBrI band gap are obtained. The electronic structure calculations were performed within GW approximation as implemented in the Vienna Ab Initio Simulation Package. The energy between lowest Eu2+ 5d state and the bottom of conduction band are found based on luminescence quenching parameters. The vacuum referred binding energy diagram of lanthanide levels was constructed using the chemical shift model.

Vacuum microelectronics for beam power and rectennas

NASA Technical Reports Server (NTRS)

Gray, Henry F.

1989-01-01

Vacuum Microelectronic devices can be described as vacuum transistors or micro-miniature vacuum tubes, as one chooses. The fundamental reason behind this new technology is the very large current densities available from field emitters, namely as high as 10(8) A/sq cm. Array current densities as high as 1000 A/sq cm have been measured. Total electron transit times from source to drain for 1 micron feature size devices have been predicted to be about 150fs. This very short transit time implies the possibility of submillimeter wave transmitters and rectennas in devices which can operate with reasonably high voltages and which are small in size and are lightweight. In addition, they are expected to be extremely radiation hard and very temperature insensitive. That is, they are expected to have radiation hardness characteristics similar to vacuum tubes, and both the high temperature and low temperature limits should be determined by the package. That is, there should be no practical intrinsic temperature or carrier freezeout problems for devices based on metals or composites. But the technology is difficult to implement at the present time because it is based on 300 to 500 angstrom radius field emitters which must be relatively uniform. There is also the need to understand the non-equilibrium transport physics in the near-surface regions of the field emitters.

Halon Replacement Program for Aviation, Aircraft Engine Nacelle Application Phase II - Operational Comparison of Selected Extinguishants

DTIC Science & Technology

1997-05-01

Control Butterfly Hi-Pressure High Flow Control Butterfly Ejector Primary Clycol Control Valve Scrubber Fan Pressure Control Butterfly 8" Venturi ...the scrubber . 20 ■ SCRUBBER FAN BLOWER INLET VALVE VP-2 VP-3 VP-4 VP-5 VP-6 VP-7 VP-8 VP-9 VP-10 SV-1 SV-2 DESCRIPTION Atmospheric...Blower Bypass Butterfly 24" Venturi Control Butterfly 24" Test Section Exit Butterfly Ejector 10’ Secondary Inlet-Butterfly Hi-pressure Low Flow

System Engineering and Integration Contract for Implementation of the National Airspace System Plan. Volume 2. Section 5.0 NAS Plan Project Findings.

DTIC Science & Technology

1984-08-01

and FPS-60 -’ vacuum-tube radars. There will be remote control interface units ( RCIU ) for tube-type radars, and an ARSR-3 relocation and RIH package...Facility RCE Remote Control Equipment; Radio Control Equipment RCIU Remote Control Interface Units RCO Remote Communications Outlet RDCC Research

Radiation dose reduction using 100-kVp and a sinogram-affirmed iterative reconstruction algorithm in adolescent head CT: Impact on grey-white matter contrast and image noise.

PubMed

Nagayama, Yasunori; Nakaura, Takeshi; Tsuji, Akinori; Urata, Joji; Furusawa, Mitsuhiro; Yuki, Hideaki; Hirarta, Kenichiro; Kidoh, Masafumi; Oda, Seitaro; Utsunomiya, Daisuke; Yamashita, Yasuyuki

2017-07-01

To retrospectively evaluate the image quality and radiation dose of 100-kVp scans with sinogram-affirmed iterative reconstruction (IR) for unenhanced head CT in adolescents. Sixty-nine patients aged 12-17 years underwent head CT under 120- (n = 34) or 100-kVp (n = 35) protocols. The 120-kVp images were reconstructed with filtered back-projection (FBP), 100-kVp images with FBP (100-kVp-F) and sinogram-affirmed IR (100-kVp-S). We compared the effective dose (ED), grey-white matter (GM-WM) contrast, image noise, and contrast-to-noise ratio (CNR) between protocols in supratentorial (ST) and posterior fossa (PS). We also assessed GM-WM contrast, image noise, sharpness, artifacts, and overall image quality on a four-point scale. ED was 46% lower with 100- than 120-kVp (p < 0.001). GM-WM contrast was higher, and image noise was lower, on 100-kVp-S than 120-kVp at ST (p < 0.001). CNR of 100-kVp-S was higher than of 120-kVp (p < 0.001). GM-WM contrast of 100-kVp-S was subjectively rated as better than of 120-kVp (p < 0.001). There were no significant differences in the other criteria between 100-kVp-S and 120-kVp (p = 0.072-0.966). The 100-kVp with sinogram-affirmed IR facilitated dramatic radiation reduction and better GM-WM contrast without increasing image noise in adolescent head CT. • 100-kVp head CT provides 46% radiation dose reduction compared with 120-kVp. • 100-kVp scanning improves subjective and objective GM-WM contrast. • Sinogram-affirmed IR decreases head CT image noise, especially in supratentorial region. • 100-kVp protocol with sinogram-affirmed IR is suited for adolescent head CT.

Modeling and optimization of sensory changes and shelf-life in vacuum-packaged cooked ham treated by E-beam irradiation

NASA Astrophysics Data System (ADS)

Benedito, J.; Cambero, M. I.; Ortuño, C.; Cabeza, M. C.; Ordoñez, J. A.; de la Hoz, L.

2011-03-01

The E-beam irradiation of vacuum-packaged RTE cooked ham was carried out to establish the dose required to achieve the food safety objective (FSO) and to minimize changes in selected sensory attributes. Cooked ham was irradiated with doses ranging 1-4 kGy. After the treatment, the microbial inactivation of Listeria monocytogenes, the shelf-life of the product and some sensory attributes (appearance, odor, and flavor) were determined. The inactivation of L. monocytogenes was satisfactorily described by a first-order kinetics equation ( R2=0.99). The influence of the irradiation dose on appearance, odor, and flavor was modeled through Gompertz ( R2=0.99, for appearance) and Activation/Inactivation ( R2=0.99, for odor and flavor) equations. A model was also developed to determine the shelf-life of irradiated cooked ham depending on the irradiation dose ( R2>0.91). The dose that maximized the scores of the sensory attributes was 0.96 kGy resulting in an acceptable sensory quality for 80 days. It is possible to apply up to 2 kGy to ensure microbial safety, while provoking no significant changes in the above mentioned sensory attributes.

Antimicrobial activity of several herb and spice extracts in culture medium and in vacuum-packaged pork.

PubMed

Kong, Baohua; Wang, Jinzhi; Xiong, Youling L

2007-03-01

Extracts prepared from honeysuckle, Scutellaria, Forsythia suspensa (Thunb), cinnamon, and rosemary with 75% ethanol and from clove oil dissolved in 75% ethanol were applied to inoculated agar media to observe their inhibitory effects on the growth of Escherichia coli, Pseudomonas fluorescens, and Lactobacillus plantarum. All the extracts suppressed the growth of these bacteria; Scutellaria exhibited the strongest effect against E. coli. An orthogonal test revealed that the most effective antimicrobial composite extracts were equal-volume mixtures of 0.125 g/ml Scutellaria + 0.5 g/ml honeysuckle + 0.125 g/ml Forsythia + 0.25 g/ml cinnamon and 0.25 g/ml cinnamon + 0.125 g/ml rosemary + 0.25% clove oil. These mixed extracts also produced strong antimicrobial effects in vacuum-packaged fresh pork, with 1.81- to 2.32-log reductions in microbial counts compared with the control when stored for up to 28 days. The sensory panel detected minimal differences in surface color and off-odors between meat samples treated with herb-spice extracts and the control. These results indicate that combined herb and spice extracts can be used as natural antimicrobials for food preservation.

Sodium chloride-induced filamentation and alternative gene expression of fts, murZ, and gnd in Listeria monocytogenes 08-5923 on vacuum-packaged ham.

PubMed

Liu, Xiaoji; Miller, Petr; Basu, Urmila; McMullen, Lynn M

2014-11-01

The aim of this study was to examine the filament formation and differential gene expression of Listeria monocytogenes 08-5923 grown on refrigerated vacuum-packaged ham products with various NaCl concentrations. Filament formation of L. monocytogenes was observed on ham products with 1.35% and 2.35% NaCl, which was monitored using flow cytometry by measuring forward light scatter. Quantitative real-time PCR was used to study the differential expression of genes in filamented cells of L. monocytogenes grown on hams following 2 or 3 months of storage at 4 °C. The genes involved in cell division (ftsX/lmo2506), cell wall synthesis (murZ/lmo2552), and NADPH production (gnd/lmo1376) were significantly downregulated in filamented cells of L. monocytogenes grown on ham with 2.35% NaCl stored at 4 °C. To our knowledge, this study reports the first evidence of filament formation of Listeria grown on meat products, which could impact the food safety risk and tolerance levels of L. monocytogenes set by regulatory agencies. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

Design and fabrication of a foldable 3D silicon based package for solid state lighting applications

NASA Astrophysics Data System (ADS)

Sokolovskij, R.; Liu, P.; van Zeijl, H. W.; Mimoun, B.; Zhang, G. Q.

2015-05-01

Miniaturization of solid state lighting (SSL) luminaires as well as reduction of packaging and assembly costs are of prime interest for the SSL lighting industry. A novel silicon based LED package for lighting applications is presented in this paper. The proposed design consists of 5 rigid Si tiles connected by flexible polyimide hinges with embedded interconnects (ICs). Electrical, optical and thermal characteristics were taken into consideration during design. The fabrication process involved polyimide (PI) application and patterning, aluminium interconnect integration in the flexible hinge, LED reflector cavity formation and metalization followed by through wafer DRIE etching for chip formation and release. A method to connect chip front to backside without TSVs was also integrated into the process. Post-fabrication wafer level assembly included LED mounting and wirebond, phosphor-based colour conversion and silicone encapsulation. The package formation was finalized by vacuum assisted wrapping around an assembly structure to form a 3D geometry, which is beneficial for omnidirectional lighting. Bending tests were performed on the flexible ICs and optical performance at different temperatures was evaluated. It is suggested that 3D packages can be expanded to platforms for miniaturized luminaire applications by combining monolithic silicon integration and system-in-package (SiP) technologies.

Potential mechanisms of carbon monoxide and high oxygen packaging in maintaining color stability of different bovine muscles.

PubMed

Liu, Chenglong; Zhang, Yimin; Yang, Xiaoyin; Liang, Rongrong; Mao, Yanwei; Hou, Xu; Lu, Xiao; Luo, Xin

2014-06-01

The objectives were to compare the effects of packaging methods on color stability, metmyoglobin-reducing-activity (MRA), total-reducing-activity and NADH concentration of different bovine muscles and to explore potential mechanisms in the enhanced color stability by carbon monoxide modified atmosphere packaging (CO-MAP, 0.4% CO/30% CO2/69.6% N2). Steaks from longissimus lumborum (LL), psoas major (PM) and longissimus thoracis (LT) packaged in CO-MAP, high-oxygen modified atmosphere packaging (HiOx-MAP, 80% O2/20% CO2) or vacuum packaging were stored for 0day, 4days, 9days, and 14days or stored for 9days then displayed in air for 0day, 1day, or 3days. The CO-MAP significantly increased red color stability of all muscles, and especially for PM. The PM and LT were more red than LL in CO-MAP, whereas PM had lowest redness in HiOx-MAP. The content of MetMb in CO-MAP was lower than in HiOx-MAP. Steaks in CO-MAP maintained a higher MRA compared with those in HiOx-MAP during storage. After opening packages, the red color of steaks in CO-MAP deteriorated more slowly compared with that of steaks in HiOx-MAP. Copyright © 2014 Elsevier Ltd. All rights reserved.

Anisotropic imaging performance in breast tomosynthesis

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

Badano, Aldo; Kyprianou, Iacovos S.; Jennings, Robert J.

We describe the anisotropy in imaging performance caused by oblique x-ray incidence in indirect detectors for breast tomosynthesis based on columnar scintillator screens. We use MANTIS, a freely available combined x-ray, electron, and optical Monte Carlo transport package which models the indirect detection processes in columnar screens, interaction by interaction. The code has been previously validated against published optical distributions. In this article, initial validation results are provided concerning the blur for particular designs of phosphor screens for which some details with respect to the columnar geometry are available from scanning electron microscopy. The polyenergetic x-ray spectrum utilized comes frommore » a database of experimental data for three different anode/filter/kVp combinations: Mo/Mo at 28 kVp, Rh/Rh at 28 kVp, and W/Al at 42 kVp. The x-ray spectra were then filtered with breast tissue (3, 4, and 6 cm thickness), compression paddle, and support base, according to the oblique paths determined by the incidence angle. The composition of the breast tissue was 50%/50% adipose/glandular tissue mass ratio. Results are reported on the pulse-height statistics of the light output and on spatial blur, expressed as the response of the detector to a pencil beam with a certain incidence angle. Results suggest that the response is nonsymmetrical and that the resolution properties of a tomosynthesis system vary significantly with the angle of x-ray incidence. In contrast, it is found that the noise due to the variability in the number of light photons detected per primary x-ray interaction changes only a few percent. The anisotropy in the response is not less in screens with absorptive backings while the noise introduced by variations in the depth-dependent light output and optical transport is larger. The results suggest that anisotropic imaging performance across the detector area can be incorporated into reconstruction algorithms for improving the image quality of breast tomosynthesis. This study also demonstrates that the assessment of image quality of breast tomosynthesis systems requires a more complete description of the detector response beyond local, center measurements of resolution and noise that assume some degree of symmetry in the detector performance.« less

Vacuum packaging of InGaAs focal plane array with four-stage thermoelectric cooler

NASA Astrophysics Data System (ADS)

Mo, De-feng; Liu, Da-fu; Yang, Li-yi; Xu, Qin-fei; Li, Xue

2013-09-01

The InGaAs focal plane array (FPA) detectors, covering the near-infrared 1~2.4 μm wavelength range, have been developed for application in space-based spectroscopy of the Earth atmosphere. This paper shows an all-metal vacuum package design for area array InGaAs detector of 1024×64 pixels, and its architecture will be given. Four-stage thermoelectric cooler (TEC) is used to cool down the FPA chip. To acquire high heat dissipation for TEC's Joule-heat, tungsten copper (CuW80) and kovar (4J29) is used as motherboard and cavity material respectively which joined by brazing. The heat loss including conduction, convection and radiation is analyzed. Finite element model is established to analyze the temperature uniformity of the chip substrate which is made of aluminum nitride (AlN). The performance of The TEC with and without heat load in vacuum condition is tested. The results show that the heat load has little influence to current-voltage relationship of TEC. The temperature difference (ΔT) increases as the input current increases. A linear relationship exists between heat load and ΔT of the TEC. Theoretical analysis and calculation show that the heat loss of radiation and conduction is about 187 mW and 82 mW respectively. Considering the Joule-heat of readout circuit and the heat loss of radiation and conduction, the FPA for a 220 K operation at room temperature can be achieved. As the thickness of AlN chip substrate is thicker than 1 millimeter, the temperature difference can be less than 0.3 K.

Formation of Covalently Modified Folding Intermediates of Simian Virus 40 Vp1 in Large T Antigen-Expressing Cells

PubMed Central

Watanabe, Marika; Phamduong, Ellen; Huang, Chu-Han; Itoh, Noriko; Bernal, Janie; Nakanishi, Akira; Rundell, Kathleen; Gjoerup, Ole

2013-01-01

The folding and pentamer assembly of the simian virus 40 (SV40) major capsid protein Vp1, which take place in the infected cytoplasm, have been shown to progress through disulfide-bonded Vp1 folding intermediates. In this report, we further demonstrate the existence of another category of Vp1 folding or assembly intermediates: the nonreducible, covalently modified mdVp1s. These species were present in COS-7 cells that expressed a recombinant SV40 Vp1, Vp1ΔC, through plasmid transfection. The mdVp1s persisted under cell and lysate treatment and SDS-PAGE conditions that are expected to have suppressed the formation of artifactual disulfide cross-links. As shown through a pulse-chase analysis, the mdVp1s were derived from the newly synthesized Vp1ΔC in the same time frame as Vp1's folding and oligomerization. The apparent covalent modifications occurred in the cytoplasm within the core region of Vp1 and depended on the coexpression of the SV40 large T antigen (LT) in the cells. Analogous covalently modified species were found with the expression of recombinant polyomavirus Vp1s and human papillomavirus L1s in COS-7 cells. Furthermore, the mdVp1s formed multiprotein complexes with LT, Hsp70, and Hsp40, and a fraction of the largest mdVp1, md4, was disulfide linked to the unmodified Vp1ΔC. Both mdVp1 formation and most of the multiprotein complex formation were blocked by a Vp1 folding mutation, C87A-C254A. Our observations are consistent with a role for LT in facilitating the folding process of SV40 Vp1 by stimulating certain covalent modifications of Vp1 or by recruiting certain cellular proteins. PMID:23427157

Vaccination with multimeric recombinant VP28 induces high protection against white spot syndrome virus in shrimp.

PubMed

Taengchaiyaphum, Suparat; Nakayama, Hideki; Srisala, Jiraporn; Khiev, Ratny; Aldama-Cano, Diva January; Thitamadee, Siripong; Sritunyalucksana, Kallaya

2017-11-01

To improve the efficacy of WSSV protection, multimeric (tetrameric) recombinant VP28 (4XrVP28) was produced and tested in comparison with those of monomeric VP28 (1XrVP28). In vitro binding of either 1XrVP28 or 4XrVP28 to shrimp hemocyte surface was evident as early as 10 min after protein inoculation. Similar results were obtained in vivo when shrimp were injected with recombinant proteins that the proteins bound to the hemocyte surface could be detected since 5 min after injection. Comparison of the WSSV protection efficiencies of 1XrVP28 or 4XrVP28 were performed by injection the purified 1XrVP28 or 4XrVP28 (22.5 μg/shrimp) and WSSV inoculum (1000 copies/shrimp) into shrimp. At 10 dpi, while shrimp injected with WSSV inoculum reached 100% mortality, shrimp injected with 1XrVP28 + WSSV or 4XrVP28 + WSSV showed relative percent survival (RPS) of 67% and 81%, respectively. PCR quantification revealed high number of WSSV in the moribund shrimp of WSSV- and 1XrVP28+WSSV-injected group. In contrast, lower number of WSSV copies were found in the survivors both from 1XrVP28+WSSV- or 4XrVP28+WSSV- injected groups. Histopathological analysis demonstrated the WSSV infected lesions found in the moribund from WSSV-infected group and 1XrVP28+WSSV-injected group, but less or none in the survivors. ELISA demonstrated that 4XrVP28 exhibited higher affinity binding to rPmRab7, a WSSV binding protein essential for WSSV entry to the cell than 1XrVP28. Taken together, the protection against WSSV in shrimp could be improved by application of multimeric rVP28. Copyright © 2017 Elsevier Ltd. All rights reserved.

[Escherichia coli heat-labile enterotoxin B subunit enhances the immune response against canine parvovirus VP2 in mice immunized by VP2 DNA vaccine].

PubMed

Han, Dongmei; Zhong, Fei; Li, Xiujin; Wang, Wei; Wang, Xingxing; Pan, Sumin

2011-01-01

To investigate the effect of Escherichia coli heat-labile enterotoxin (LT) B subunit (LTB) gene on canine parvovirus (CPV) VP2 gene vaccine. The LTB gene was amplified by PCR from genomic DNA of E. coli 44815 strain. The VP2-70 fragment (210 bp) encoding major epitope of VP2 (70 amino acids) was amplified by PCR from a plasmid encoding VP2 gene. VP2-70 and LTB genes were inserted into the eukaryotic vector to construct VP2-70 gene,LTB gene and VP2-70-LTB fused gene vectors. The mice were immunized with VP2-70 vector, VP2-70-LTB fused vector, or VP2-70 vector plus LTB vector, respectively. The antibody titers at the different time were measured by using ELISA method. The spleen lymphocyte proliferation activity was analyzed by 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. The sequence of VP2-70 and LTB genes was identified. The recombinant VP2-70 and LTB proteins could be expressed in HEK293T cells in a secretory manner. The mice immunized with VP2-70 vector, VP2-70-LTB vector or VP2-70 vector plus LTB vector could generate the specific antibody against VP2 protein. The antibody titer immunized with VP2-70-LTB vector reached 1:5120 at 35 d post immunization, significantly higher than that of other two groups (P < 0.01). For antibody isotype analysis, the IgG1 isotype antibody titers in all test groups were significantly higher than of IgG2a (P < 0.01). The high-level spleen lymphocyte stimulation index was observed in the three test groups under the stimulation with Con A, higher than that in control groups (P < 0.01). LTB gene could enhance the humoral immune response of CPV VP2 gene vaccine in mice.

Full protection against African horsesickness (AHS) in horses induced by baculovirus-derived AHS virus serotype 4 VP2, VP5 and VP7.

PubMed

Martínez-Torrecuadrada, J L; Díaz-Laviada, M; Roy, P; Sánchez, C; Vela, C; Sánchez-Vizcaíno, J M; Casal, J I

1996-06-01

African horsesickness virus serotype 4 (AHSV-4) outer capsid protein VP2, or VP2 and VP5 plus inner capsid protein VP7, derived from single or dual recombinant baculovirus expression vectors were used in different combinations to immunize horses. When the proteins were purified by affinity chromatography, the combination of all three proteins induced low levels of neutralizing antibodies and conferred protection against virulent virus challenge. However, purified VP2 or VP2 and VP5 in the absence of VP7 failed to induce neutralizing antibodies and protection. Immunization with non-purified proteins enhanced the titres of neutralizing antibodies. Again, the combination of the three proteins was able to confer total protection to immunized horses, which showed absence of viraemia. The antigenicity of recombinant VP2 was analysed with a collection of 30 MAbs. Both purified and unpurified recombinant VP2 proteins showed different antigenic patterns in comparison to that of VP2 on virions. An immunization experiment with four more horses confirmed these results. The vaccine described here would not only prevent the disease, but would drastically reduce the propagation of the virus by vectors.

Flexible Foam Protection Materials for Portable Life Support System Packaging Study

NASA Technical Reports Server (NTRS)

Tang,Henry H.; Dillon, Paul A.; Thomas, Gretchen A.

2009-01-01

This paper discusses the phase I effort in evaluating and selecting a light weight impact protection material for the Constellation Space Suit Element (CSSE) Portable Life Support System (PLSS) conceptual packaging study. A light weight material capable of holding and protecting the components inside the PLSS is required to demonstrate the viability of the flexible PLSS packaging concept. The material needs to distribute, dissipate, and absorb the impact energy of the PLSS falling on the lunar surface. It must also be robust to consistently perform over several Extravehicular Activity (EVA) missions in the extreme lunar thermal vacuum environment. This paper documents the performance requirements for selecting a foam protection material, and the methodologies for evaluating some commercial off-the-shelf (COTS) foam material candidates. It also presents the mechanical properties and impact drop tests results of the foam material candidates. The results of this study suggest that a foam based flexible protection system is a viable solution for PLSS packaging. However, additional works are needed to optimize COTS foam or to develop a composite foam system that will meet all the performance requirements for the CSSE PLSS flexible packaging.

Flexible Foam Protection Materials for Constellation Space Suit Element Portable Life Support Subsystem Packaging Study

NASA Technical Reports Server (NTRS)

Tang, Henry H.; Orndoff, Evelyne S.; Thomas, Gretchen A.

2009-01-01

This paper discusses the effort in evaluating and selecting a light weight impact protection material for the Constellation Space Suit Element (CSSE) Portable Life Support Subsystem (PLSS) conceptual packaging study. A light weight material capable of holding and protecting the components inside the PLSS is required to demonstrate the viability of the flexible PLSS packaging concept. The material needs to distribute, dissipate, and absorb the impact energy of the PLSS falling on the lunar surface. It must also be very robust and function in the extreme lunar thermal vacuum environment for up to one hundred Extravehicular Activity (EVA) missions. This paper documents the performance requirements for selecting a foam protection material, and the methodologies for evaluating commercial off-the-shelf (COTS) foam protection materials. It also presents the materials properties test results and impact drop test results of the various foam materials evaluated in the study. The findings from this study suggest that a foam based flexible protection system is a viable solution for PLSS packaging. However, additional works are needed to optimize COTS foam properties or to develop a composite foam system that will meet all the performance requirements for the CSSE PLSS flexible packaging.

Plasma membrane association facilitates conformational changes in the Marburg virus protein VP40 dimer.

PubMed

Bhattarai, Nisha; Gc, Jeevan B; Gerstman, Bernard S; Stahelin, Robert V; Chapagain, Prem P

2017-04-26

Filovirus infections cause hemorrhagic fever in humans and non-human primates that often results in high fatality rates. The Marburg virus is a lipid-enveloped virus from the Filoviridae family and is closely related to the Ebola virus. The viral matrix layer underneath the lipid envelope is formed by the matrix protein VP40 (VP40), which is also involved in other functions during the viral life-cycle. As in the Ebola virus VP40 (eVP40), the recently determined X-ray crystal structure of the Marburg virus VP40 (mVP40) features loops containing cationic residues that form a lipid binding basic patch. However, the mVP40 basic patch is significantly flatter with a more extended surface than in eVP40, suggesting the possibility of differences in the plasma membrane interactions and phospholipid specificity between the VP40 dimers. In this paper, we report on molecular dynamics simulations that investigate the roles of various residues and lipid types in PM association as well as the conformational changes of the mVP40 dimer facilitated by membrane association. We compared the structural changes of the mVP40 dimer with the mVP40 dimer in both lipid free and membrane associated conditions. Despite the significant structural differences in the crystal structure, the Marburg VP40 dimer is found to adopt a configuration very similar to the Ebola VP40 dimer after associating with the membrane. This conformational rearrangement upon lipid binding allows Marburg VP40 to localize and stabilize at the membrane surface in a manner similar to the Ebola VP40 dimer. Consideration of the structural information in its lipid-interacting condition may be important in targeting mVP40 for novel drugs to inhibit viral budding from the plasma membrane.

Herpes simplex virus 2 VP22 phosphorylation induced by cellular and viral kinases does not influence intracellular localization

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

Geiss, Brian J.; Cano, Gina L.; Tavis, John E.

2004-12-05

Phosphorylation of the herpes simplex virus (HSV) VP22 protein is regulated by cellular kinases and the UL13 viral kinase, but the sites at which these enzymes induce phosphorylation of HSV-2 VP22 are not known. Using serine-to-alanine mutants to map phosphorylation sites on HSV-2 VP22 in cells, we made three major observations. First, phosphorylation by a cellular kinase mapped to serines 70, 71, and/or 72 within CKII consensus sites analogous to previously identified phosphorylation sites in HSV-1 VP22. Second, we mapped UL13-mediated phosphorylation of HSV-2 VP22 to serines 28 and 34, describing for the first time UL13-dependent phosphorylation sites on VP22.more » Third, previously identified VP22-associated cellular kinase sites in HSV-1 VP22 (serines 292 and 294) were not phosphorylated in HSV-2 VP22 (serines 291 and 293). VP22 expressed alone accumulated in the cytoplasm and to a lesser extent in the nucleus. Phosphorylation by endogenous cellular kinase(s) did not alter the localization of VP22. Co-expression of HSV-2 VP22 with active UL13, but not with enzymatically inactive UL13, resulted in nuclear accumulation of VP22 and altered nuclear morphology. Surprisingly, redistribution of VP22 to the nucleus occurred independently of UL13-induced phosphorylation of VP22. The altered nuclear morphology of UL13-expressing cells was not due to apoptosis. These results demonstrate that phosphorylation of HSV-2 VP22 at multiple serine residues is induced by UL13 and cellular kinase(s), and that the nuclear/cytoplasmic distribution of VP22 is independent of its phosphorylation status but is controlled indirectly by UL13 kinase activity.« less

Expression and characterization of a novel truncated rotavirus VP4 for the development of a recombinant rotavirus vaccine.

PubMed

Li, Yijian; Xue, Miaoge; Yu, Linqi; Luo, Guoxing; Yang, Han; Jia, Lianzhi; Zeng, Yuanjun; Li, Tingdong; Ge, Shengxiang; Xia, Ningshao

2018-04-12

The outer capsid protein VP4 is an important target for the development of a recombinant rotavirus vaccine because it mediates the attachment and penetration of rotavirus. Due to the poor solubility of full-length VP4, VP8 was explored as candidate rotavirus vaccines in the past years. In previous studies, it has been found that the N-terminal truncated VP8 protein, VP8-1 (aa26-231), could be expressed in soluble form with improved immunogenicity compared to the core of VP8 (aa65-223). However, this protein stimulated only a weak immune response when aluminum hydroxide was used as an adjuvant. In addition, it should be noted that the protective efficacy of VP4 was higher than that of VP8 and VP5. In this study, it was found that when the N-terminal 25 amino acids were deleted, the truncated VP4 ∗ (aa26-476) containing VP8 and the stalk domain of VP5 could be expressed in soluble form in E. coli and purified to homogeneous trimers. Furthermore, the truncated VP4 could induce high titers of neutralizing antibodies when aluminum adjuvant was used and conferred high protective efficacy in reducing the severity of diarrhea and rotavirus shedding in stools in animal models. The immunogenicity of the truncated VP4 was significantly higher than that of VP8 ∗ and VP5 ∗ alone. Taken together, the truncated VP4 ∗ (aa26-476), with enhanced immunogenicity and immunoprotectivity, could be considered as a viable candidate for further development and has the potential to become a parenterally administered rotavirus vaccine. Copyright © 2018 Elsevier Ltd. All rights reserved.

Simulation of a turbulent supersonic underexpanded jet flowing into a submerged space with the help of a shear stress transfer model

NASA Astrophysics Data System (ADS)

Isaev, S. A.; Lipnitskii, Yu. M.; Baranov, P. A.; Panasenko, A. V.; Usachov, A. E.

2012-11-01

We have calculated the flow of an axisymmetric turbulent supersonic underexpanded jet into a submerged space with the help of the VP2/3 package as part of the generalized pressure correction procedure. The shear stress transfer model modified with account for the curvature of streamlines has been verified on the basis of comparison with V. I. Zapryagaev's data obtained at the S. A. Khristianovich Institute of Theoretical and Applied Mechanics, Siberian Branch of the Russian Academy of Sciences. The influence of the generated vortex viscosity on the shock-wave structure of the jet, the field of flow parameters, and the turbulence characteristics has been analyzed.

Effects of sous-vide method at different temperatures, times and vacuum degrees on the quality, structural, and microbiological properties of pork ham.

PubMed

Jeong, Kiyoung; O, Hyeonbin; Shin, So Yeon; Kim, Young-Soon

2018-04-10

This study evaluated the influence of different factors on pork hams cooked by sous-vide method. The quality and structural and microbiological properties of the treated samples were compared with those of controls. Samples were subjected to treatment at different combinations of temperature (61 °C or 71 °C), time (45 or 90 min), and vacuum degree (98.81% or 96.58%). The control sample was air packaged and boiled for 45 min in boiling water. Temperature and vacuum degree affected quality properties, while the effect of time was limited. Samples cooked at 61 °C showed higher moisture content, redness, and pink color of the meat juice, whereas samples cooked at 71 °C showed higher cooking loss rate, lightness, and volatile basic nitrogen values. Texture analysis indicated tenderer meat for the treatment group than the control. No microbial growth was detected in any treatment groups. Meat cooked at 61 °C and 98.81% vacuum showed more spacious arrangement of meat fiber. Copyright © 2018 Elsevier Ltd. All rights reserved.

Examinations on Laser Remote Welding of Ultra-thin Metal Foils Under Vacuum Conditions

NASA Astrophysics Data System (ADS)

Petrich, Martin; Stambke, Martin; Bergmann, Jean Pierre

Metal foils are commonly used for catalytic converters, vacuum insulations, in medical and electrical industry as well as for sensor applications and packaging. The investigations in this paper determine the influence of reduced atmospheric pressure during the welding process with a highly brilliant 400 W single-mode fiber laser combined with a 2D-scanning system. The laser beam is transmitted through a highly transparent glass into a vacuum chamber, where AISI 304 stainless steel foils with a thickness of 25 μm, 50 μm and 100 μm are positioned. The effects of reduced atmospheric pressure on the plasma formation are investigated by means of high-speed videography. Furthermore, the geometry of the weld seam is compared to atmospheric conditions as well as means of the process stability and the process efficiency. The welds were also evaluated by means of metallography. The research is a contribution for extending the range of micro welding applications and shows new aspects for future developments.

VP1 amino acid residue 145 of enterovirus 71 is a key residue for its receptor attachment and resistance to neutralizing antibody during cynomolgus monkey infection.

PubMed

Fujii, Ken; Sudaka, Yui; Takashino, Ayako; Kobayashi, Kyousuke; Kataoka, Chikako; Suzuki, Tadaki; Iwata-Yoshikawa, Naoko; Kotani, Osamu; Ami, Yasushi; Shimizu, Hiroyuki; Nagata, Noriyo; Mizuta, Katsumi; Matsuzaki, Yoko; Koike, Satoshi

2018-05-30

Enterovirus 71 (EV71) is a causative agent of hand, foot, and mouth disease and sometimes causes severe or fatal neurological complications. The amino acid at VP1-145 determines virological characteristics of EV71. Viruses with glutamic acid (E) at VP1-145 (VP1-145E) are virulent in neonatal mice and transgenic mice expressing human scavenger receptor B2, whereas those with glutamine (Q) or glycine (G) are not. However, the contribution of this variation to pathogenesis in humans is not fully understood. We compared the virulence of VP1-145E and VP1-145G viruses of Isehara and C7/Osaka backgrounds in cynomolgus monkeys. VP1-145E, but not VP1-145G, viruses induced neurological symptoms. VP1-145E viruses were frequently detected in the tissues of infected monkeys. VP1-145G viruses were detected less frequently and disappeared quickly. Instead, mutants that had a G to E mutation at VP1-145 emerged, suggesting that VP1-145E viruses have a replication advantage in the monkeys. This is consistent with our hypothesis proposed in the accompanying paper that the VP1-145G virus is attenuated due to its adsorption by heparan sulfate. Monkeys infected with both viruses produced neutralizing antibodies before the onset of the disease. Interestingly, VP1-145E viruses were more resistant to neutralizing antibodies than VP1-145G viruses in vitro A small amount of neutralizing antibody raised in the early phase of infection may not be sufficient to block the dissemination of VP1-145E viruses. The different resistance of the VP1-145 variants to neutralizing antibodies may be one of the reasons for the difference in virulence. IMPORTANCE The contribution of VP1-145 variants in humans is not fully understood. In some reports, VP1-145G/Q viruses were more frequently isolated from severely affected than from mildly affected patients, suggesting that VP1-145G/Q viruses are more virulent. In the accompanying paper, we showed that VP1-145E viruses are more virulent than VP1-145G viruses in human SCARB2 transgenic mice. Heparan sulfate acts as a decoy to specifically trap the VP1-145G viruses and leads to abortive infection. Here, we demonstrated that VP1-145G was attenuated in cynomolgus monkeys, suggesting that this hypothesis is also true in a non-human primate model. VP1-145E viruses, but not VP1-145G viruses, were highly resistant to neutralizing antibodies. We propose the difference in resistance against neutralizing antibodies as another mechanism of EV71 virulence. In summary, VP1-145 contributes to virulence determination by controlling attachment receptor usage and antibody sensitivity. Copyright © 2018 American Society for Microbiology.

Consumer preferences for beef color and packaging did not affect eating satisfaction.

PubMed

Carpenter, C E; Cornforth, D P; Whittier, D

2001-04-01

We investigated whether consumer preferences for beef colors (red, purple, and brown) or for beef packaging systems (modified atmosphere, MAP; vacuum skin pack, VSP; or overwrap with polyvinyl chloride, PVC) influenced taste scores of beef steaks and patties. To test beef color effects, boneless beef top loin steaks (choice) and ground beef patties (20% fat) were packaged in different atmospheres to promote development of red, purple, and brown color. To test effects of package type, steaks and patties were pre-treated with carbon monoxide in MAP to promote development of red color, and some meat was repackaged using VSP or PVC overwrap. The differently colored and packaged meats were separately displayed for members of four consumer panels who evaluated appearance and indicated their likelihood to purchase similar meat. Next, the panelists tasted meat samples from what they had been told were the packaging treatments just observed. However, the meat samples actually served were from a single untreated steak or patty. Thus, any difference in taste scores should reflect expectations established during the visual evaluation. The same ballot and sample coding were used for both the visual and taste evaluations. Color and packaging influenced (P<0.001) appearance scores and likelihood to purchase. Appearance scores were rated red>purple >brown and PVC >VSP>MAP. Appearance scores and likelihood to purchase were correlated (r=0.9). However, color or packaging did not affect (P>0.5) taste scores. Thus, consumer preferences for beef color and packaging influenced likelihood to purchase, but did not bias eating satisfaction.

Behavior of Listeria monocytogenes at 7 degrees C in commercial turkey breast, with or without antimicrobials, after simulated contamination for manufacturing, retail and consumer settings.

PubMed

Lianou, Alexandra; Geornaras, Ifigenia; Kendall, Patricia A; Scanga, John A; Sofos, John N

2007-08-01

Uncured turkey breast, commercially available with or without a mixture of potassium lactate and sodium diacetate, was sliced, inoculated with a 10-strain composite of Listeria monocytogenes, vacuum-packaged, and stored at 4 degrees C, to simulate contamination after a lethal processing step at the plant. At 5, 15, 25 and 50 days of storage, packages were opened, slices were tested, and bags with remaining slices were reclosed with rubber bands; this simulated home use of plant-sliced and -packaged product. At the same above time intervals, portions of original product (stored at 4 degrees C in original processing bags) were sliced and inoculated as above, and packaged in delicatessen bags, simulating contamination during slicing/handling at retail or home. Both sets of bags were stored aerobically at 7 degrees C for 12 days to simulate home storage. L. monocytogenes populations were lower (P<0.05) during storage in turkey breast containing a combination of lactate and diacetate compared to product without antimicrobials under both contamination scenarios. Due to prolific growth of the pathogen under the plant-contamination scenario in product without lactate-diacetate during vacuum-packaged storage (4 degrees C), populations at 3 days of aerobic storage (7 degrees C) of such product ranged from 4.6 to 7.4 log cfu/cm(2). Under the retail/home-contamination scenario, mean growth rates (log cfu/cm(2)/day) of the organism during aerobic storage ranged from 0.14 to 0.16, and from 0.25 to 0.51, in product with and without lactate-diacetate, respectively; growth rates in turkey breast without antimicrobials decreased (P<0.05) with age of the product. Overall, product without antimicrobials inoculated to simulate plant-contamination and product with lactate-diacetate inoculated to simulate retail/home-contamination were associated with the highest and lowest pathogen levels during aerobic storage at 7 degrees C, respectively. However, 5- and 15-day-old turkey breast without lactate-diacetate stored aerobically for 12 days resulted in similar pathogen levels (7.3-7.7 log cfu/cm(2)), irrespective of contamination scenario.

New seismic Vp- and Vp/Vs- models of HUKKA 2007 wide-angle reflection and refraction profile in northern Fennoscandian Shield

NASA Astrophysics Data System (ADS)

Tiira, T.; Janik, T.; Kozlovskaya, E.; Grad, M.; Korja, A.; Komminaho, K.; Hegedüs, E.; Kovács, C. A.; Silvennoinen, H.; Brückl, E.

2012-04-01

We study the block structure within accreationary orogens. We present an example from northern part of the Fennoscandian Shield transected by deep seismic sounding profile HUKKA 2007. The 455 km long profile runs in NNW-SSE direction from Kittilä in northwestern Finnish Lapland to Kostamush in Russia near central part of the border between Finland and Russia. We present 2-D seismic velocity model (Vp and Vp/Vs ratio in the crust, depth to the Moho and depth to the intracrustal reflectors) along HUKKA 2007 wide-angle reflection and refraction profile in northern Finland. Commercial and military chemical explosions at 7 shot points were used as sources of the seismic energy. The shots were recorded by 115 recording stations deployed along the profile with an average station spacing of 3.45 km. The field recordings were cut and sorted into shot gathers. The 2-D velocity model of the HUKKA 2007 profile was developed by SEIS83 forward raytracing package using arrivals of major refracted and reflected P- and S-wave phases. In general the velocities vary in the upper crust between 5.8 and 6.1 km/s. Interesting features are three high P wave velocity (6.30-6.35 km/s) bodies in the upper crust. Two small bodies lie close to surface at first 100 km and the third one can be followed from 200 to 350 km along the profile reaching depth of 5-10 km. The central part of the profile (between 120 and 220 km) has a zone of low (lower than 6 km/s) P-wave velocity in the uppermost crust. This zone is about 4 km thick. In addition, the velocity model along the HUKKA 2007 profile shows significant difference in crustal velocity structure between the northern (up to 120 km) and southern parts of the profile. The differences in P-wave velocities and Vp/Vs ratio can be followed throughout the crust down to the Moho boundary. This suggests that the HUKKA 2007 profile transects a major terrane boundary. However, the position of this boundary with respect to major crustal units is controversial. It may be the boundary that separates the pristine parts of the Archean Karelian craton from those parts reworked in the Paleoproterozoic. Alternatively, it can be the boundary that separates the Karelian craton from the Belomorian mobile belt.

T helper cell-mediated interferon-gamma expression after human parvovirus B19 infection: persisting VP2-specific and transient VP1u-specific activity

PubMed Central

Franssila, R; Auramo, J; Modrow, S; Möbs, M; Oker-Blom, C; Käpylä, P; Söderlund-Venermo, M; Hedman, K

2005-01-01

Human parvovirus B19 is a small non-enveloped DNA virus with an icosahedral capsid consisting of proteins of only two species, the major protein VP2 and the minor protein VP1. VP2 is contained within VP1, which has an additional unique portion (VP1u) of 227 amino acids. We determined the ability of eukaryotically expressed parvovirus B19 virus-like particles consisting of VP1 and VP2 in the ratio recommended for vaccine use, or of VP2 alone, to stimulate, in an HLA class II restricted manner, peripheral blood mononuclear cells (PBMC) to proliferate and to secrete interferon gamma (IFN-γ) and interleukin (IL)-10 cytokines among recently and remotely B19 infected subjects. PBMC reactivity with VP1u was determined specifically with a prokaryotically expressed VP1u antigen. In general, B19-specific IFN-γ responses were stronger than IL-10 responses in both recent and remote infection; however, IL-10 responses were readily detectable among both groups, with the exception of patients with relapsed or persisting symptoms who showed strikingly low IL-10 responses. Whereas VP1u-specific IFN-γ responses were very strong among the recently infected subjects, the VP1u-specific IFN-γ and IL-10 responses were virtually absent among the remotely infected subjects. The disappearance of VP1u-specific IFN-γ expression is surprising, as B-cell immunity against VP1u is well maintained. PMID:16178856

Lloviu virus VP24 and VP35 proteins function as innate immune antagonists in human and bat cells

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

Feagins, Alicia R.; Basler, Christopher F., E-mail: chris.basler@mssm.edu

Lloviu virus (LLOV) is a new member of the filovirus family that also includes Ebola virus (EBOV) and Marburg virus (MARV). LLOV has not been cultured; however, its genomic RNA sequence indicates the coding capacity to produce homologs of the EBOV and MARV VP24, VP35, and VP40 proteins. EBOV and MARV VP35 proteins inhibit interferon (IFN)-alpha/beta production and EBOV VP35 blocks activation of the antiviral kinase PKR. The EBOV VP24 and MARV VP40 proteins inhibit IFN signaling, albeit by different mechanisms. Here we demonstrate that LLOV VP35 suppresses Sendai virus induced IFN regulatory factor 3 (IRF3) phosphorylation, IFN-α/β production, andmore » PKR phosphorylation. Additionally, LLOV VP24 blocks tyrosine phosphorylated STAT1 binding to karyopherin alpha 5 (KPNA5), STAT1 nuclear accumulation, and IFN-induced gene expression. LLOV VP40 lacks detectable IFN antagonist function. These activities parallel EBOV IFN inhibitory functions. EBOV and LLOV VP35 and VP24 proteins also inhibit IFN responses in bat cells. These data suggest that LLOV infection will block innate immune responses in a manner similar to EBOV. - Highlights: • Lloviu virus (LLOV) is a new member of the filovirus family. • LLOV VP35 blocks IRF3 phosphorylation, IFN-α/β production and PKR phosphorylation. • LLOV VP24 inhibits IFN responses by targeting phospho-STAT1 KPNA interaction. • Infection by LLOV may block innate immune responses in a manner similar to EBOV.« less

Effect of high-dose irradiation on quality characteristics of ready-to-eat broiler breast fillets stored at room temperature.

PubMed

Baptista, R F; Teixeira, C E; Lemos, M; Monteiro, M L G; Vital, H C; Mársico, E T; Júnior, C A Conte; Mano, S B

2014-10-01

The effect of high-dose irradiation on the physical, chemical, and bacteriological parameters of ready-to-eat vacuum-packed broiler breast meat after 430 d of storage at room temperature was investigated. Ready-to-eat broiler breast fillets were immersed in brine with garlic powder and then drained, grilled, and vacuum-packed (primary packaging). The high-dose irradiation used was approximately 48 kGy. The treatments were designated as A (irradiated samples stored at room temperature), B (irradiated samples stored at -25°C), and C (nonirradiated samples stored at -25°C). All samples were packaged in polyethylene bags containing aluminum to exclude light (secondary packaging). Proximate composition, pH, 2-thiobarbituric acid reactive substance (TBARS), and heterotrophic aerobic mesophilic bacteria were analyzed during 430 d of storage. Results were analyzed using 1-way ANOVA and the Tukey test. Linear regression was used to analyze the correlation between the results for each parameter and storage time of the different treatments. The gamma radiation caused slight changes (P < 0.05) in the moisture and fat content, regardless of storage temperature. After storage d 110, TBARS values remained stable (P > 0.05) in all the treatments. The preservation methods used were effective in maintaining the mesophilic counts below the detection level during the entire storage period. We concluded that, among the treatments studied, high-dose irradiation with storage at room temperature showed potential for the preservation of ready-to-eat products made from poultry meat, to provide foods safe for consumption. ©2014 Poultry Science Association Inc.

Practical application of HgI2 detectors to a space-flight scanning electron microscope

NASA Technical Reports Server (NTRS)

Bradley, J. G.; Conley, J. M.; Albee, A. L.; Iwanczyk, J. S.; Dabrowski, A. J.

1989-01-01

Mercuric iodide X-ray detectors have been undergoing tests in a prototype scanning electron microscope system being developed for unmanned space flight. The detector program addresses the issues of geometric configuration in the SEM, compact packaging that includes separate thermoelectric coolers for the detector and FET, X-ray transparent hermetic encapsulation and electrical contacts, and a clean vacuum environment.

Pre-harvest sprouting resistance and haplotype variation of ThVp-1 gene in the collection of wheat-wheatgrass hybrids

PubMed Central

Kocheshkova, A. A.; Kroupin, P. Yu.; Bazhenov, M. S.; Karlov, G. I.; Pochtovyy, A. A.; Upelniek, V. P.; Belov, V. I.

2017-01-01

The germplasm collection of 87 wheat-wheatgrass hybrids developed in Tsitisin Main Botanical Garden (Russia, Moscow) was evaluated for resistance to pre-harvest sprouting (PHS) by spike sprouting (SS) and germination index (GI) assays as well as for spike and grain features. The PHS resistance variation and haplotype polymorphism of the wheatgrass ThVp-1 and wheat TaVp-1B genes orthologues of Vp-1 was revealed in the studied collection. Four haplotypes of ThVp-1 were revealed: ThVp-1a (41% of the entries), ThVp-1b (13%), ThVp-1c (29%), and ThVp-1d (15%). The association between the allelic state of ThVp-1 and PHS resistance in the wheat-wheatgrass hybrids was shown: haplotype ThVp-1d of the wheatgrass Vp-1 gene is significantly associated with reduced PHS in the wheat-wheatgrass hybrids (mean SS 0.33, mean GI 0.64). The resistant entries may be perspective as a source of PHS resistance in the development of commercial cultivars of perennial wheat. PMID:29131854

Cyclophilin A Interacts with Viral VP4 and Inhibits the Replication of Infectious Bursal Disease Virus.

PubMed

Wang, Nian; Zhang, Lizhou; Chen, Yuming; Lu, Zhen; Gao, Li; Wang, Yongqiang; Gao, Yulong; Gao, Honglei; Cui, Hongyu; Li, Kai; Liu, Changjun; Zhang, Yanping; Qi, Xiaole; Wang, Xiaomei

2015-01-01

Nonstructural protein VP4, a serine protease of infectious bursal disease virus (IBDV) that catalyzes the hydrolysis of polyprotein pVP2-VP4-VP3 to form the viral proteins VP2, VP4, and VP3, is essential to the replication of IBDV. However, the interacting partners of VP4 in host cells and the effects of the interaction on the IBDV lifecycle remain incompletely elucidated. In this study, using the yeast two-hybrid system, the putative VP4-interacting partner cyclophilin A (CypA) was obtained from a chicken embryo fibroblast (CEF) expression library. CypA was further confirmed to interact with VP4 of IBDV using co-immunoprecipitation (CO-IP), GST pull-down, and confocal microscopy assays. Moreover, we found that the overexpression of CypA suppressed IBDV replication, whereas the knock-down of CypA by small interfering RNAs promoted the replication of IBDV. Taken together, our findings indicate that the host cell protein CypA interacts with viral VP4 and inhibits the replication of IBDV.

Biochemical and Functional Characterization of the Ebola Virus VP24 Protein: Implications for a Role in Virus Assembly and Budding

PubMed Central

Han, Ziying; Boshra, Hani; Sunyer, J. Oriol; Zwiers, Susan H.; Paragas, Jason; Harty, Ronald N.

2003-01-01

The VP24 protein of Ebola virus is believed to be a secondary matrix protein and minor component of virions. In contrast, the VP40 protein of Ebola virus is the primary matrix protein and the most abundant virion component. The structure and function of VP40 have been well characterized; however, virtually nothing is known regarding the structure and function of VP24. Wild-type and mutant forms of VP24 were expressed in mammalian cells to gain a better understanding of the biochemical and functional nature of this viral protein. Results from these experiments demonstrated that (i) VP24 localizes to the plasma membrane and perinuclear region in both transfected and Ebola virus-infected cells, (ii) VP24 associates strongly with lipid membranes, (iii) VP24 does not contain N-linked sugars when expressed alone in mammalian cells, (iv) VP24 can oligomerize when expressed alone in mammalian cells, (v) progressive deletions at the N terminus of VP24 resulted in a decrease in oligomer formation and a concomitant increase in the formation of high-molecular-weight aggregates, and (vi) VP24 was present in trypsin-resistant virus like particles released into the media covering VP24-transfected cells. These data indicate that VP24 possesses structural features commonly associated with viral matrix proteins and that VP24 may have a role in virus assembly and budding. PMID:12525613

Computed tomographic venography for varicose veins of the lower extremities: prospective comparison of 80-kVp and conventional 120-kVp protocols.

PubMed

Cho, Eun-Suk; Kim, Joo Hee; Kim, Sungjun; Yu, Jeong-Sik; Chung, Jae-Joon; Yoon, Choon-Sik; Lee, Hyeon-Kyeong; Lee, Kyung Hee

2012-01-01

To prospectively investigate the feasibility of an 80-kilovolt (peak) (kVp) protocol in computed tomographic venography for varicose veins of the lower extremities by comparison with conventional 120-kVp protocol. Attenuation values and signal-to-noise ratio of iodine contrast medium (CM) were determined in a water phantom for 2 tube voltages (80 kVp and 120 kVp). Among 100 patients, 50 patients were scanned with 120 kVp and 150 effective milliampere second (mAs(eff)), and the other 50 patients were scanned with 80 kVp and 390 mAs(eff) after the administration of 1.7-mL/kg CM (370 mg of iodine per milliliter). The 2 groups were compared for venous attenuation, contrast-to-noise ratio, and subjective degree of venous enhancement, image noise, and overall diagnostic image quality. In the phantom, the attenuation value and signal-to-noise ratio value for iodine CM at 80 kVp were 63.8% and 33.0% higher, respectively, than those obtained at 120 kVp. The mean attenuation of the measured veins of the lower extremities was 148.3 Hounsfield units (HU) for the 80-kVp protocol and 94.8 HU for the 120-kVp protocol. Contrast-to-noise ratio was also significantly higher with the 80-kVp protocol. The overall diagnostic image quality of the 3-dimensional volume-rendered images was good with both protocols. The subjective score for venous enhancement was higher at the 80-kVp protocol. The mean volume computed tomography dose index of the 80-kVp (5.6 mGy) protocol was 23.3% lower than that of the 120-kVp (7.3 mGy) protocol. The use of the 80-kVp protocol improved overall venous attenuation, especially in perforating vein, and provided similarly high diagnostic image quality with a lower radiation dose when compared to the conventional 120-kVp protocol.

Assembly of the Herpes Simplex Virus Capsid: Preformed Triplexes Bind to the Nascent Capsid

PubMed Central

Spencer, Juliet V.; Newcomb, William W.; Thomsen, Darrell R.; Homa, Fred L.; Brown, Jay C.

1998-01-01

The herpes simplex virus type 1 (HSV-1) capsid is a T=16 icosahedral shell that forms in the nuclei of infected cells. Capsid assembly also occurs in vitro in reaction mixtures created from insect cell extracts containing recombinant baculovirus-expressed HSV-1 capsid proteins. During capsid formation, the major capsid protein, VP5, and the scaffolding protein, pre-VP22a, condense to form structures that are extended into procapsids by addition of the triplex proteins, VP19C and VP23. We investigated whether triplex proteins bind to the major capsid-scaffold protein complexes as separate polypeptides or as preformed triplexes. Assembly products from reactions lacking one triplex protein were immunoprecipitated and examined for the presence of the other. The results showed that neither triplex protein bound unless both were present, suggesting that interaction between VP19C and VP23 is required before either protein can participate in the assembly process. Sucrose density gradient analysis was employed to determine the sedimentation coefficients of VP19C, VP23, and VP19C-VP23 complexes. The results showed that the two proteins formed a complex with a sedimentation coefficient of 7.2S, a value that is consistent with formation of a VP19C-VP232 heterotrimer. Furthermore, VP23 was observed to have a sedimentation coefficient of 4.9S, suggesting that this protein exists as a dimer in solution. Deletion analysis of VP19C revealed two domains that may be required for attachment of the triplex to major capsid-scaffold protein complexes; none of the deletions disrupted interaction of VP19C with VP23. We propose that preformed triplexes (VP19C-VP232 heterotrimers) interact with major capsid-scaffold protein complexes during assembly of the HSV-1 capsid. PMID:9557680

Hepatitis A Virus Capsid Protein VP1 Has a Heterogeneous C Terminus

PubMed Central

Graff, Judith; Richards, Oliver C.; Swiderek, Kristine M.; Davis, Michael T.; Rusnak, Felicia; Harmon, Shirley A.; Jia, Xi-Yu; Summers, Donald F.; Ehrenfeld, Ellie

1999-01-01

Hepatitis A virus (HAV) encodes a single polyprotein which is posttranslationally processed into the functional structural and nonstructural proteins. Only one protease, viral protease 3C, has been implicated in the nine protein scissions. Processing of the capsid protein precursor region generates a unique intermediate, PX (VP1-2A), which accumulates in infected cells and is assumed to serve as precursor to VP1 found in virions, although the details of this reaction have not been determined. Coexpression in transfected cells of a variety of P1 precursor proteins with viral protease 3C demonstrated efficient production of PX, as well as VP0 and VP3; however, no mature VP1 protein was detected. To identify the C-terminal amino acid residue of HAV VP1, we performed peptide sequence analysis by protease-catalyzed [18O]H2O incorporation followed by liquid chromatography ion-trap microspray tandem mass spectrometry of HAV VP1 isolated from purified virions. Two different cell culture-adapted isolates of HAV, strains HM175pE and HM175p35, were used for these analyses. VP1 preparations from both virus isolates contained heterogeneous C termini. The predominant C-terminal amino acid in both virus preparations was VP1-Ser274, which is located N terminal to a methionine residue in VP1-2A. In addition, the analysis of HM175pE recovered smaller amounts of amino acids VP1-Glu273 and VP1-Thr272. In the case of HM175p35, which contains valine at amino acid position VP1-273, VP1-Thr272 was found in addition to VP1-Ser274. The data suggest that HAV 3C is not the protease responsible for generation of the VP1 C terminus. We propose the involvement of host cell protease(s) in the production of HAV VP1. PMID:10364353

Localization of the herpes simplex virus type 1 major capsid protein VP5 to the cell nucleus requires the abundant scaffolding protein VP22a.

PubMed

Nicholson, P; Addison, C; Cross, A M; Kennard, J; Preston, V G; Rixon, F J

1994-05-01

The intracellular distributions of three herpes simplex virus type 1 (HSV-1) capsid proteins, VP23, VP5 and VP22a, were examined using vaccinia virus and plasmid expression systems. During infection of cells with HSV-1 wild-type virus, all three proteins were predominantly located in the nucleus, which is the site of capsid assembly. However, when expressed in the absence of any other HSV-1 proteins, although VP22a was found exclusively in the nucleus as expected, VP5 and VP23 were distributed throughout the cell. Thus nuclear localization is not an intrinsic property of these proteins but must be mediated by one or more HSV-1-induced proteins. Co-expression experiments demonstrated that VP5 was efficiently transported to the nucleus in the presence of VP22a, but the distribution of VP23 was unaffected by the presence of either or both of the other two proteins.

Characterization of putative toxin/antitoxin systems in Vibrio parahaemolyticus.

PubMed

Hino, M; Zhang, J; Takagi, H; Miyoshi, T; Uchiumi, T; Nakashima, T; Kakuta, Y; Kimura, M

2014-07-01

To obtain more information about the toxin/antitoxin (TA) systems in the Vibrio genus and also to examine their involvement in the induction of a viable but nonculturable (VBNC) state, we searched homologues of the Escherichia coli TA systems in the Vibrio parahaemolyticus genome. We found that a gene cluster, vp1842/vp1843, in the V. parahaemolyticus genome database has homology to that encoding the E. coli TA proteins, DinJ/YafQ. Expression of the putative toxin gene vp1843 in E. coli cells strongly inhibited the cell growth, while coexpression with the putative antitoxin gene vp1842 neutralized this effect. Mutational analysis identified Lys37 and Pro45 in the gene product VP1843 of vp1843 as crucial residues for the growth retardation of E. coli cells. VP1843, unlike the E. coli toxin YafQ, has no protein synthesis inhibitory activity, and that instead the expression of vp1843 in E. coli caused morphological change of the cells. The gene cluster vp1842/vp1843 encodes the V. parahaemolyticus TA system; VP1843 inhibits cell growth, whereas VP1842 serves as an antitoxin by forming a stable complex with VP1843. The putative toxin, VP1843, may be involved in the induction of the VBNC state in V. parahaemolyticus by inhibiting cell division. © 2014 The Society for Applied Microbiology.

Color, lipid oxidation, sensory quality, and aroma compounds of beef steaks displayed under different levels of oxygen in a modified atmosphere package.

PubMed

Resconi, Virginia Celia; Escudero, Ana; Beltrán, José Antonio; Olleta, José Luis; Sañudo, Carlos; Campo, María del Mar

2012-01-01

High oxygen modified atmosphere packaging (MAP) is currently used by the industry to maintain an attractive color in fresh meat. However, it can also promote lipid oxidation and sensory changes. The aim of this study was to compare the quality of beef steaks displayed under different levels of oxygen. For that purpose, meat was stored with 50%, 60%, and 80% of oxygen for 4 and 8 d at retail conditions. A control treatment with steaks vacuum packaged, without exposure to retail display, was included, and color, lipid oxidation, organoleptic characteristics, and aroma compounds were analyzed. Steaks displayed under high oxygen MAP with 50% of oxygen showed the lowest color stability, according to hue values. Higher oxygen levels did not necessarily correspond with higher rancidity levels in the raw meat. Thiobarbituric Acid Reacting Substances (TBARS) values were higher than 2.2 mg malonaldehyde (MDA)/kg muscle in all the samples displayed for 8 d at retail conditions. These samples exhibited the highest intensities of rancid and the least intense beef odors and flavors. The ketones: 2,3-butanedione, 2-octanone, 2,3-pentanedione, 2-heptanone, 4-methyl-2-pentanone, and the aldehydes: pentanal, 2-methyl-butanal, and 2-furfurylthiol, 1-octen-3-ol and 2-methylpropyl-acetate, were proposed as candidates for the aroma differences between the cooked beef steaks stored under vacuum and high oxygen packs. High oxygen modified atmosphere packaging (MAP) is used to increase shelf-life and color at the retail level. However, oxygen deteriorates faster some quality attributes, being a contributor to lipid oxidation that could be perceived at consumption. This study attempts to find the best gas composition in a MAP in order to maintain the color, minimizing the oxidation. Aromatically relevant chemicals have been analyzed by gas olfactometry-gas chromatography as a key to measure beef oxidation after display. © 2011 Institute of Food Technologists®

Qualitative and quantitative evaluation of rigid and deformable motion correction algorithms using dual-energy CT images in view of application to CT perfusion measurements in abdominal organs affected by breathing motion

PubMed Central

Skornitzke, S; Fritz, F; Klauss, M; Pahn, G; Hansen, J; Hirsch, J; Grenacher, L; Kauczor, H-U

2015-01-01

Objective: To compare six different scenarios for correcting for breathing motion in abdominal dual-energy CT (DECT) perfusion measurements. Methods: Rigid [RRComm(80 kVp)] and non-rigid [NRComm(80 kVp)] registration of commercially available CT perfusion software, custom non-rigid registration [NRCustom(80 kVp], demons algorithm) and a control group [CG(80 kVp)] without motion correction were evaluated using 80 kVp images. Additionally, NRCustom was applied to dual-energy (DE)-blended [NRCustom(DE)] and virtual non-contrast [NRCustom(VNC)] images, yielding six evaluated scenarios. After motion correction, perfusion maps were calculated using a combined maximum slope/Patlak model. For qualitative evaluation, three blinded radiologists independently rated motion correction quality and resulting perfusion maps on a four-point scale (4 = best, 1 = worst). For quantitative evaluation, relative changes in metric values, R2 and residuals of perfusion model fits were calculated. Results: For motion-corrected images, mean ratings differed significantly [NRCustom(80 kVp) and NRCustom(DE), 3.3; NRComm(80 kVp), 3.1; NRCustom(VNC), 2.9; RRComm(80 kVp), 2.7; CG(80 kVp), 2.7; all p < 0.05], except when comparing NRCustom(80 kVp) with NRCustom(DE) and RRComm(80 kVp) with CG(80 kVp). NRCustom(80 kVp) and NRCustom(DE) achieved the highest reduction in metric values [NRCustom(80 kVp), 48.5%; NRCustom(DE), 45.6%; NRComm(80 kVp), 29.2%; NRCustom(VNC), 22.8%; RRComm(80 kVp), 0.6%; CG(80 kVp), 0%]. Regarding perfusion maps, NRCustom(80 kVp) and NRCustom(DE) were rated highest [NRCustom(80 kVp), 3.1; NRCustom(DE), 3.0; NRComm(80 kVp), 2.8; NRCustom(VNC), 2.6; CG(80 kVp), 2.5; RRComm(80 kVp), 2.4] and had significantly higher R2 and lower residuals. Correlation between qualitative and quantitative evaluation was low to moderate. Conclusion: Non-rigid motion correction improves spatial alignment of the target region and fit of CT perfusion models. Using DE-blended and DE-VNC images for deformable registration offers no significant improvement. Advances in knowledge: Non-rigid algorithms improve the quality of abdominal CT perfusion measurements but do not benefit from DECT post processing. PMID:25465353

Two potential recombinant rabies vaccines expressing canine parvovirus virion protein 2 induce immunogenicity to canine parvovirus and rabies virus.

PubMed

Luo, Jun; Shi, Hehe; Tan, Yeping; Niu, Xuefeng; Long, Teng; Zhao, Jing; Tian, Qin; Wang, Yifei; Chen, Hao; Guo, Xiaofeng

2016-08-17

Both rabies virus (RABV) and canine parvovirus (CPV) cause lethal diseases in dogs. In this study, both high egg passage Flury (HEP-Flury) strains of RABV and recombinant RABV carrying double RABV glycoprotein (G) gene were used to express the CPV virion protein 2 (VP2) gene, and were designated rHEP-VP2 and, rHEP-dG-VP2 respectively. The two recombinant RABVs maintained optimal virus titration according to their viral growth kinetics assay compared with the parental strain HEP-Flury. Western blotting indicated that G protein and VP2 were expressed in vitro. The expression of VP2 in Crandell feline kidney cells post-infection by rHEP-VP2 and rHEP-dG-VP2 was confirmed by indirect immunofluorescence assay with antibody against VP2. Immunogenicity of recombinant rabies viruses was tested in Kunming mice. Both rHEP-VP2 and rHEP-dG-VP2 induced high levels of rabies antibody compared with HEP-Flury. Mice immunized with rHEP-VP2 and rHEP-dG-VP2 both had a high level of antibodies against VP2, which can protect against CPV infection. A challenge experiment indicated that more than 80% mice immunized with recombinant RABVs survived after infection of challenge virus standard 24 (CVS-24). Together, this study showed that recombinant RABVs expressing VP2 induced protective immune responses to RABV and CPV. Therefore, rHEP-VP2 and rHEP-dG-VP2 might be potential combined vaccines for RABV and CPV. Copyright © 2016 Elsevier Ltd. All rights reserved.

Enterovirus 71 viral capsid protein linear epitopes: Identification and characterization

PubMed Central

2012-01-01

Background To characterize the human humoral immune response against enterovirus 71 (EV71) infection and map human epitopes on the viral capsid proteins. Methods A series of 256 peptides spanning the capsid proteins (VP1, VP2, VP3) of BJ08 strain (genomic C4) were synthesized. An indirect enzyme-linked immunosorbent assay (ELISA) was carried out to detect anti-EV71 IgM and IgG in sera of infected children in acute or recovery phase. The partially overlapped peptides contained 12 amino acids and were coated in the plate as antigen (0.1 μg/μl). Sera from rabbits immunized with inactivated BJ08 virus were also used to screen the peptide panel. Results A total of 10 human anti-EV71 IgM epitopes (vp1-14 in VP1; vp2-6, 21, 40 and 50 in VP2 and vp3-10, 12, 15, 24 and 75 in VP3) were identified in acute phase sera. In contrast, only one anti-EV71 IgG epitope in VP1 (vp1-15) was identified in sera of recovery stage. Four rabbit anti-EV71 IgG epitopes (vp1-14, 31, 54 and 71) were identified and mapped to VP1. Conclusion These data suggested that human IgM epitopes were mainly mapped to VP2 and VP3 with multi-epitope responses occurred at acute infection, while the only IgG epitope located on protein VP1 was activated in recovery phase sera. The dynamic changes of humoral immune response at different stages of infection may have public health significance in evaluation of EV71 vaccine immunogenicity and the clinical application of diagnostic reagents. PMID:22264266

WE-FG-202-12: Investigation of Longitudinal Salivary Gland DCE-MRI Changes

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

Ger, R; Howell, R; Li, H

Purpose: To determine the correlation between dose and changes through treatment in dynamic contrast enhanced (DCE) MRI voxel parameters (Ktrans, kep, Ve, and Vp) within salivary glands of head and neck oropharyngeal squamous cell carcinoma (HNSCC) patients. Methods: 17 HNSCC patients treated with definitive radiation therapy completed DCE-MRI scans on a 3T scanner at pre-treatment, mid-treatment, and post-treatment time points. Mid-treatment and post-treatment DCE images were deformably registered to pre-treatment DCE images (Velocity software package). Pharmacokinetic analysis of the DCE images used a modified Tofts model to produce parameter maps with an arterial input function selected from each patient’s perivertebralmore » space on the image (NordicICE software package). In-house software was developed for voxel-by-voxel longitudinal analysis of the salivary glands within the registered images. The planning CT was rigidly registered to the pre-treatment DCE image to obtain dose values in each voxel. Voxels within the lower and upper dose quartiles for each gland were averaged for each patient, then an average of the patients’ means for the two quartiles were compared. Dose-relationships were also assessed by Spearman correlations between dose and voxel parameter changes for each patient’s gland. Results: Changes in parameters’ means between time points were observed, but inter-patient variability was high. Ve of the parotid was the only parameter that had a consistently significant longitudinal difference between dose quartiles. The highest Spearman correlation was Vp of the sublingual gland for the change in the pre-treatment to mid-treatment values with only a ρ=0.29. Conclusion: In this preliminary study, there was large inter-patient variability in the changes of DCE voxel parameters with no clear relationship with dose. Additional patients may reduce the uncertainties and allow for the determination of the existence of parameter and dose relationships.« less

Overexpression of VpEIFP1, a novel F-box/Kelch-repeat protein from wild Chinese Vitis pseudoreticulata, confers higher tolerance to powdery mildew by inducing thioredoxin z proteolysis.

PubMed

Wang, Jie; Yao, Wenkong; Wang, Lei; Ma, Fuli; Tong, Weihuo; Wang, Chen; Bao, Rui; Jiang, Changyue; Yang, Yazhou; Zhang, Jianxia; Xu, Yan; Wang, Xiping; Zhang, Chaohong; Wang, Yuejin

2017-10-01

An F-box protein (VpEIFP1) induced by Erysiphe necator was isolated from Vitis pseudoreticulata, a wild Chinese grapevine species naturally resistant to powdery mildew (PM). It contains an F-box domain and two Kelch-repeat motifs. Expression profiles indicate the VpEIFP1 is strongly induced at both transcriptional and translational levels by PM infection. A subcellular localisation assay showed that VpEIFP1 is predominantly located in the nucleus and cytoplasm. Overexpression of VpEIFP1 accelerated the accumulation of hydrogen peroxide (H 2 O 2 ) and up-regulated the expressions of ICS2, NPR1 and PR1 involved in defence responses, resulting in suppression of PM germination and growth. As an F-box protein, VpEIFP1 interacts with thioredoxin z (VpTrxz) in the yeast-two-hybrid (Y2H) assay and in the bimolecular fluorescence complementation (BiFC) assay. Decreased amounts of VpTrxz protein in transgenic grapevine leaves overexpressing VpEIFP1 were restored by proteasome inhibitor MG132, implying that VpEIFP1 mediated VpTrxz for degradation through the SCF VpEIFP1 (Skp1-Cullin-F-box) E3 ubiquitin ligase complex. The RNA interference line of VpTrxz showed increased H 2 O 2 accumulation following PM inoculation. We propose VpEIFP1 positively modulates the grapevine defence response to PM by inducing the degradation of VpTrxz via the ubiquitin/26S proteasome system. Copyright © 2017 Elsevier B.V. All rights reserved.

The immunogenicity of recombinant vaccines based on modified Vaccinia Ankara (MVA) viruses expressing African horse sickness virus VP2 antigens depends on the levels of expressed VP2 protein delivered to the host.

PubMed

Calvo-Pinilla, Eva; Gubbins, Simon; Mertens, Peter; Ortego, Javier; Castillo-Olivares, Javier

2018-06-01

African horse sickness (AHS) is a lethal equine disease transmitted by Culicoides biting midges and caused by African horse sickness virus (AHSV). AHS is endemic to sub-Saharan Africa, but devastating outbreaks have been recorded periodically outside this region. The perceived risk of an AHS outbreak occurring in Europe has increased following the frequent epidemics caused in ruminants by bluetongue virus, closely related to AHSV. Attenuated vaccines for AHS are considered unsuitable for use in non-endemic countries due bio-safety concerns. Further, attenuated and inactivated vaccines are not compatible with DIVA (differentiate infected from vaccinated animals) strategies. All these factors stimulated the development of novel AHS vaccines that are safer, more efficacious and DIVA compatible. We showed previously that recombinant modified Vaccinia Ankara virus (MVA) vaccines encoding the outer capsid protein of AHSV (AHSV-VP2) induced virus neutralising antibodies (VNAb) and protection against AHSV in a mouse model and also in the horse. Passive immunisation studies demonstrated that immunity induced by MVA-VP2 was associated with pre-challenge VNAb titres in the vaccinates. Analyses of the inoculum of these MVA-VP2 experimental vaccines showed that they contained pre-formed AHSV-VP2. We continued studying the influence of pre-formed AHSV-VP2, present in the inoculum of MVA-VP2 vaccines, in the immunogenicity of MVA-VP2 vaccines. Thus, we compared correlates of immunity in challenged mice that were previously vaccinated with: a) MVA-VP2 (live); b) MVA-VP2 (live and sucrose gradient purified); c) MVA-VP2 (UV light inactivated); d) MVA-VP2 (UV light inactivated and diluted); e) MVA-VP2 (heat inactivated); f) MVA-VP2 (UV inactivated) + MVA-VP2 (purified); g) MVA-VP2 (heat inactivated) + MVA-VP2 (purified); and h) wild type-MVA (no insert). The results of these experiments showed that protection was maximal using MVA-VP2 (live) vaccine and that the protection conferred by all other vaccines correlated strongly with the levels of pre-formed AHSV-VP2 in the vaccine inoculum. Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

Proteomic analysis reveals differential accumulation of small heat shock proteins and late embryogenesis abundant proteins between ABA-deficient mutant vp5 seeds and wild-type Vp5 seeds in maize

PubMed Central

Wu, Xiaolin; Gong, Fangping; Yang, Le; Hu, Xiuli; Tai, Fuju; Wang, Wei

2014-01-01

ABA is a major plant hormone that plays important roles during many phases of plant life cycle, including seed development, maturity and dormancy, and especially the acquisition of desiccation tolerance. Understanding of the molecular basis of ABA-mediated plant response to stress is of interest not only in basic research on plant adaptation but also in applied research on plant productivity. Maize mutant viviparous-5 (vp5), deficient in ABA biosynthesis in seeds, is a useful material for studying ABA-mediated response in maize. Due to carotenoid deficiency, vp5 endosperm is white, compared to yellow Vp5 endosperm. However, the background difference at proteome level between vp5 and Vp5 seeds is unclear. This study aimed to characterize proteome alterations of maize vp5 seeds and to identify ABA-dependent proteins during seed maturation. We compared the embryo and endosperm proteomes of vp5 and Vp5 seeds by gel-based proteomics. Up to 46 protein spots, most in embryos, were found to be differentially accumulated between vp5 and Vp5. The identified proteins included small heat shock proteins (sHSPs), late embryogenesis abundant (LEA) proteins, stress proteins, storage proteins and enzymes among others. However, EMB564, the most abundant LEA protein in maize embryo, accumulated in comparable levels between vp5 and Vp5 embryos, which contrasted to previously characterized, greatly lowered expression of emb564 mRNA in vp5 embryos. Moreover, LEA proteins and sHSPs displayed differential accumulations in vp5 embryos: six out of eight identified LEA proteins decreased while nine sHSPs increased in abundance. Finally, we discussed the possible causes of global proteome alterations, especially the observed differential accumulation of identified LEA proteins and sHSPs in vp5 embryos. The data derived from this study provides new insight into ABA-dependent proteins and ABA-mediated response during maize seed maturation. PMID:25653661

Dynamic Phosphorylation of VP30 Is Essential for Ebola Virus Life Cycle.

PubMed

Biedenkopf, Nadine; Lier, Clemens; Becker, Stephan

2016-05-15

Ebola virus is the causative agent of a severe fever with high fatality rates in humans and nonhuman primates. The regulation of Ebola virus transcription and replication currently is not well understood. An important factor regulating viral transcription is VP30, an Ebola virus-specific transcription factor associated with the viral nucleocapsid. Previous studies revealed that the phosphorylation status of VP30 impacts viral transcription. Together with NP, L, and the polymerase cofactor VP35, nonphosphorylated VP30 supports viral transcription. Upon VP30 phosphorylation, viral transcription ceases. Phosphorylation weakens the interaction between VP30 and the polymerase cofactor VP35 and/or the viral RNA. VP30 thereby is excluded from the viral transcription complex, simultaneously leading to increased viral replication which is supported by NP, L, and VP35 alone. Here, we use an infectious virus-like particle assay and recombinant viruses to show that the dynamic phosphorylation of VP30 is critical for the cotransport of VP30 with nucleocapsids to the sites of viral RNA synthesis, where VP30 is required to initiate primary viral transcription. We further demonstrate that a single serine residue at amino acid position 29 was sufficient to render VP30 active in primary transcription and to generate a recombinant virus with characteristics comparable to those of wild-type virus. In contrast, the rescue of a recombinant virus with a single serine at position 30 in VP30 was unsuccessful. Our results indicate critical roles for phosphorylated and dephosphorylated VP30 during the viral life cycle. The current Ebola virus outbreak in West Africa has caused more than 28,000 cases and 11,000 fatalities. Very little is known regarding the molecular mechanisms of how the Ebola virus transcribes and replicates its genome. Previous investigations showed that the transcriptional support activity of VP30 is activated upon VP30 dephosphorylation. The current study reveals that the situation is more complex and that primary transcription as well as the rescue of recombinant Ebola virus also requires the transient phosphorylation of VP30. VP30 encodes six N-proximal serine residues that serve as phosphorylation acceptor sites. The present study shows that the dynamic phosphorylation of serine at position 29 alone is sufficient to activate primary viral transcription. Our results indicate a series of phosphorylation/dephosphorylation events that trigger binding to and release from the nucleocapsid and transcription complex to be essential for the full activity of VP30. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Sexual dimorphism of human vallate papillae: an in vivo study of normative morphology.

PubMed

Zdilla, M J; Gibson, L N; Hunt, A J

2015-01-01

The perimeters of vallate papillae (VP) house approximately half of the taste buds on the human tongue. However, little information exists regarding perimeter measurements of VP. Likewise, great diversity exists among reports of the number of VP and diameter of VP, in general. The research presents an analysis of the perimeters, counts, and diameters of VP in vivo. Endoscopic examination was performed on 79 individuals (40 females, 39 males) between 18 and 26 years of age. A total of 583 VP were counted, 565 of which were able to be measured. Data revealed a statistically significant difference between male and female VP count (t(75.6) = 4.5; p = 0.00003). Females had, on average, 2.22 more VP than males. Males were found to have larger mean VP diameter per person and mean VP perimeter per person than females (t(58.9) = -2.4; p = 0.021 and t(59.3) = -2.4; p = 0.019, respectively). The report demonstrates that VP are sexually dimorphic at the gross anatomical level.

Independent segregation of two antigenic specificities (VP3 and VP7) involved in neutralization of rotavirus infectivity.

PubMed Central

Hoshino, Y; Sereno, M M; Midthun, K; Flores, J; Kapikian, A Z; Chanock, R M

1985-01-01

Antiserum prepared against the M37 strain of rotavirus, recovered from an asymptomatic newborn infant in Venezuela, neutralized two prototype human rotaviruses that define two separate serotypes: serotype 1 (Wa) and serotype 4 (ST3). Thus, the M37 strain is a naturally occurring intertypic rotavirus. Analysis of reassortant viruses produced during coinfection in vitro indicated that the observed dual serotype specificity of M37 resulted from sharing a related outer capsid protein, VP3, with the ST3 virus and another related outer capsid protein, VP7, with the Wa virus. Analysis of single (VP3)-gene-substitution reassortants indicated that VP3 was as potent an immunogen as VP7. In addition, direct evidence was obtained that the serotype specificity of neutralizing antibody elicited by VP3 can differ from the serotype specificity of neutralizing antibody elicited by VP7, indicating the need for a dual system of rotavirus classification in which the neutralization specificity of both VP3 and VP7 outer capsid proteins are identified. Images PMID:3001716

Aerogel-Based Multilayer Insulation with Micrometeoroid Protection

NASA Technical Reports Server (NTRS)

Begag, Redouane; White, Shannon

2013-01-01

Ultra-low-density, highly hydrophobic, fiber-reinforced aerogel material integrated with MLI (aluminized Mylar reflectors and B4A Dacron separators) offers a highly effective insulation package by providing unsurpassed thermal performance and significant robustness, delivering substantial MMOD protection via the addition of a novel, durable, external aerogel layer. The hydrophobic nature of the aerogel is an important property for maintaining thermal performance if the material is exposed to the environment (i.e. rain, snow, etc.) during ground installations. The hybrid aerogel/MLI/MMOD solution affords an attractive alternative because it will perform thermally in the same range as MLI at all vacuum levels (including high vacuum), and offers significant protection from micrometeoroid damage. During this effort, the required low-density and resilient aerogel materials have been developed that are needed to optimize the thermal performance for space (high vacuum) cryotank applications. The proposed insulation/MMOD package is composed of two sections: a stack of interleaved aerogel layers and MLI intended for cryotank thermal insulation, and a 1.5- to 1-in. (.2.5- to 3.8- cm) thick aerogel layer (on top of the insulation portion) for MMOD protection. Learning that low-density aerogel cannot withstand the hypervelocity impact test conditions, the innovators decided during the course of the program to fabricate a high-density and strong material based on a cross-linked aerogel (X-aerogel; developed elsewhere by the innovators) for MMOD protection. This system has shown a very high compressive strength that is capable of withstanding high-impact tests if a proper configuration of the MMOD aerogel layer is used. It was learned that by stacking two X-aerogel layers [1.5-in. (.3.8-cm) thick] separated by an air gap, the system would be able to hold the threat at a speed of 5 km/s and gpass h the test. The first aerogel panel stopped the projectile from damaging the second aerogel panel. The impacted X-aerogel (the back specimen from the successful test) was further tested in comparison to another similar sample (not impacted) at Kennedy Space Center for thermal conductivity evaluation at cryogenic conditions. The specimens were tested under high vacuum and cryogenic temperatures, using Cryostat 500. The results show that the specimen did not lose a significant amount of thermal performance due to the impact test, especially at high vacuum.

The chilled storage life and retail display performance of vacuum and carbon dioxide packed hot deboned beef striploins.

PubMed

Bell, R G; Penney, N; Gilbert, K V; Moorhead, S M; Scott, S M

1996-04-01

Two cooling regimes that complied with the New Zealand meat hygiene requirement that hot deboned meat be chilled to +7 °C or less within 24 hr of leaving the slaughter floor were evaluated for the production of chilled table meats. Electrically stimulated hot deboned bull beef half striploins were either vacuum or carbon dioxide packed before being cooled in accordance with either Regime 1 (cool at +5 °C for 24 hr, transfer to chiller operating at -1.0 ± 0.5 °C) or Regime 2 (cool at +5 °C for 24 hr, hold at 5 °C for 6 days, transfer to chiller operating at -1.0 ± 0.5 °C). Striploins were removed from -1.0 °C storage 8, 28, 42, 56, 70, 84 and 98 days after slaughter and subjected to microbiological, tenderness, sensory and retail display performance evaluations. Both Regimes 1 and 2 produced meat of acceptable mean tenderness, 8 kgF (MIRINZ Tenderometer) in either vacuum or carbon dioxide packs within 28 and 8 days of slaughter, respectively. However, 70 days after slaughter the first signs of over-ageing became apparent. Steaks from Regimes 1 and 2 maintained acceptable visual appearance during retail display at 5 °C for 48 hr and 24 hr, respectively. After these times, the product was judged by the panel to be unacceptable because of its dull dark lean tissue and grey to green discoloration of the fat. Poor colour stability during retail display was mirrored by deterioration of sensory attributes, particularly aroma which is indicative of incipient spoilage. While carbon dioxide packaging in combination with Regime 1 offered an initial microbiological advantage over vacuum packaging, this advantage was not, however, carried over into retail display. Poor colour and sensory stability during retail display suggest that chilled table cuts derived from hot deboned bull beef are more suited to the Hotel-Restaurant-Institutional (HRI) trade than supermarket retailing. To serve the HRI, vacuum packed hot deboned bull beef primal cuts processed by Regime 1 appear to be the combination of choice. This combination would enable commercial processors to produce quality table beef with a chilled storage life of up to 70 days.

Remote sensing measurements of sea surface temperature as an indicator of Vibrio parahaemolyticus in oyster meat and human illnesses.

PubMed

Konrad, Stephanie; Paduraru, Peggy; Romero-Barrios, Pablo; Henderson, Sarah B; Galanis, Eleni

2017-08-31

Vibrio parahaemolyticus (Vp) is a naturally occurring bacterium found in marine environments worldwide. It can cause gastrointestinal illness in humans, primarily through raw oyster consumption. Water temperatures, and potentially other environmental factors, play an important role in the growth and proliferation of Vp in the environment. Quantifying the relationships between environmental variables and indicators or incidence of Vp illness is valuable for public health surveillance to inform and enable suitable preventative measures. This study aimed to assess the relationship between environmental parameters and Vp in British Columbia (BC), Canada. The study used Vp counts in oyster meat from 2002-2015 and laboratory confirmed Vp illnesses from 2011-2015 for the province of BC. The data were matched to environmental parameters from publicly available sources, including remote sensing measurements of nighttime sea surface temperature (SST) obtained from satellite readings at a spatial resolution of 1 km. Using three separate models, this paper assessed the relationship between (1) daily SST and Vp counts in oyster meat, (2) weekly mean Vp counts in oysters and weekly Vp illnesses, and (3) weekly mean SST and weekly Vp illnesses. The effects of salinity and chlorophyll a were also evaluated. Linear regression was used to quantify the relationship between SST and Vp, and piecewise regression was used to identify SST thresholds of concern. A total of 2327 oyster samples and 293 laboratory confirmed illnesses were included. In model 1, both SST and salinity were significant predictors of log(Vp) counts in oyster meat. In model 2, the mean log(Vp) count in oyster meat was a significant predictor of Vp illnesses. In model 3, weekly mean SST was a significant predictor of weekly Vp illnesses. The piecewise regression models identified a SST threshold of approximately 14 o C for both model 1 and 3, indicating increased risk of Vp in oyster meat and Vp illnesses at higher temperatures. Monitoring of SST, particularly through readily accessible remote sensing data, could serve as a warning signal for Vp and help inform the introduction and cessation of preventative or control measures.

Expression of goose parvovirus whole VP3 protein and its epitopes in Escherichia coli cells.

PubMed

Tarasiuk, K; Woźniakowski, G; Holec-Gąsior, L

2015-01-01

The aim of this study was the expression of goose parvovirus capsid protein (VP3) and its epitopes in Escherichia coli cells. Expression of the whole VP3 protein provided an insufficient amount of protein. In contrast, the expression of two VP3 epitopes (VP3ep4, VP3ep6) in E. coli, resulted in very high expression levels. This may suggest that smaller parts of the GPV antigenic determinants are more efficiently expressed than the complete VP3 gene.

Folding units in calcium vector protein of amphioxus: Structural and functional properties of its amino- and carboxy-terminal halves.

PubMed

Baladi, S; Tsvetkov, P O; Petrova, T V; Takagi, T; Sakamoto, H; Lobachov, V M; Makarov, A A; Cox, J A

2001-04-01

Muscle of amphioxus contains large amounts of a four EF-hand Ca2+-binding protein, CaVP, and its target, CaVPT. To study the domain structure of CaVP and assess the structurally important determinants for its interaction with CaVPT, we expressed CaVP and its amino (N-CaVP) and carboxy-terminal halves (C-CaVP). The interactive properties of recombinant and wild-type CaVP are very similar, despite three post-translational modifications in the wild-type protein. N-CaVP does not bind Ca2+, shows a well-formed hydrophobic core, and melts at 44 degrees C. C-CaVP binds two Ca2+ with intrinsic dissociation constants of 0.22 and 140 microM (i.e., very similar to the entire CaVP). The metal-free domain in CaVP and C-CaVP shows no distinct melting transition, whereas its 1Ca2+ and 2Ca2+) forms melt in the 111 degrees -123 degrees C range, suggesting that C-CaVP and the carboxy- domain of CaVP are natively unfolded in the metal-free state and progressively gain structure upon binding of 1Ca2+ and 2Ca2+. Thermal denaturation studies provide evidence for interdomain interaction: the apo, 1Ca2+ and 2Ca2+ states of the carboxy-domain destabilize to different degrees the amino-domain. Only C-CaVP forms a Ca2+-dependent 1:1 complex with CaVPT. Our results suggest that the carboxy-terminal domain of CaVP interacts with CaVPT and that the amino-terminal lobe modulates this interaction.

Folding units in calcium vector protein of amphioxus: Structural and functional properties of its amino- and carboxy-terminal halves

PubMed Central

Baladi, Sibyl; Tsvetkov, Philipp O.; Petrova, Tatiana V.; Takagi, Takashi; Sakamoto, Hiroshi; Lobachov, Vladimir M.; Makarov, Alexander A.; Cox, Jos A.

2001-01-01

Muscle of amphioxus contains large amounts of a four EF-hand Ca2+-binding protein, CaVP, and its target, CaVPT. To study the domain structure of CaVP and assess the structurally important determinants for its interaction with CaVPT, we expressed CaVP and its amino (N-CaVP) and carboxy-terminal halves (C-CaVP). The interactive properties of recombinant and wild-type CaVP are very similar, despite three post-translational modifications in the wild-type protein. N-CaVP does not bind Ca2+, shows a well-formed hydrophobic core, and melts at 44°C. C-CaVP binds two Ca2+ with intrinsic dissociation constants of 0.22 and 140 μM (i.e., very similar to the entire CaVP). The metal-free domain in CaVP and C-CaVP shows no distinct melting transition, whereas its 1Ca2+ and 2Ca2+ forms melt in the 111°–123°C range, suggesting that C-CaVP and the carboxy- domain of CaVP are natively unfolded in the metal-free state and progressively gain structure upon binding of 1Ca2+ and 2Ca2+. Thermal denaturation studies provide evidence for interdomain interaction: the apo, 1Ca2+ and 2Ca2+ states of the carboxy-domain destabilize to different degrees the amino-domain. Only C-CaVP forms a Ca2+-dependent 1:1 complex with CaVPT. Our results suggest that the carboxy-terminal domain of CaVP interacts with CaVPT and that the amino-terminal lobe modulates this interaction. PMID:11274468

Crystal Structures of Yellowtail Ascites Virus VP4 Protease

PubMed Central

Chung, Ivy Yeuk Wah; Paetzel, Mark

2013-01-01

Yellowtail ascites virus (YAV) is an aquabirnavirus that causes ascites in yellowtail, a fish often used in sushi. Segment A of the YAV genome codes for a polyprotein (pVP2-VP4-VP3), where processing by its own VP4 protease yields the capsid protein precursor pVP2, the ribonucleoprotein-forming VP3, and free VP4. VP4 protease utilizes the rarely observed serine-lysine catalytic dyad mechanism. Here we have confirmed the existence of an internal cleavage site, preceding the VP4/VP3 cleavage site. The resulting C-terminally truncated enzyme (ending at Ala716) is active, as shown by a trans full-length VP4 cleavage assay and a fluorometric peptide cleavage assay. We present a crystal structure of a native active site YAV VP4 with the internal cleavage site trapped as trans product complexes and trans acyl-enzyme complexes. The acyl-enzyme complexes confirm directly the role of Ser633 as the nucleophile. A crystal structure of the lysine general base mutant (K674A) reveals the acyl-enzyme and empty binding site states of VP4, which allows for the observation of structural changes upon substrate or product binding. These snapshots of three different stages in the VP4 protease reaction mechanism will aid in the design of anti-birnavirus compounds, provide insight into previous site-directed mutagenesis results, and contribute to understanding of the serine-lysine dyad protease mechanism. In addition, we have discovered that this protease contains a channel that leads from the enzyme surface (adjacent to the substrate binding groove) to the active site and the deacylating water. PMID:23511637

Crystal structures of yellowtail ascites virus VP4 protease: trapping an internal cleavage site trans acyl-enzyme complex in a native Ser/Lys dyad active site.

PubMed

Chung, Ivy Yeuk Wah; Paetzel, Mark

2013-05-03

Yellowtail ascites virus (YAV) is an aquabirnavirus that causes ascites in yellowtail, a fish often used in sushi. Segment A of the YAV genome codes for a polyprotein (pVP2-VP4-VP3), where processing by its own VP4 protease yields the capsid protein precursor pVP2, the ribonucleoprotein-forming VP3, and free VP4. VP4 protease utilizes the rarely observed serine-lysine catalytic dyad mechanism. Here we have confirmed the existence of an internal cleavage site, preceding the VP4/VP3 cleavage site. The resulting C-terminally truncated enzyme (ending at Ala(716)) is active, as shown by a trans full-length VP4 cleavage assay and a fluorometric peptide cleavage assay. We present a crystal structure of a native active site YAV VP4 with the internal cleavage site trapped as trans product complexes and trans acyl-enzyme complexes. The acyl-enzyme complexes confirm directly the role of Ser(633) as the nucleophile. A crystal structure of the lysine general base mutant (K674A) reveals the acyl-enzyme and empty binding site states of VP4, which allows for the observation of structural changes upon substrate or product binding. These snapshots of three different stages in the VP4 protease reaction mechanism will aid in the design of anti-birnavirus compounds, provide insight into previous site-directed mutagenesis results, and contribute to understanding of the serine-lysine dyad protease mechanism. In addition, we have discovered that this protease contains a channel that leads from the enzyme surface (adjacent to the substrate binding groove) to the active site and the deacylating water.

Tandem truncated rotavirus VP8* subunit protein with T cell epitope as non-replicating parenteral vaccine is highly immunogenic.

PubMed

Wen, Xiaobo; Cao, Dianjun; Jones, Ronald W; Hoshino, Yasutaka; Yuan, Lijuan

2015-01-01

The two currently available live oral rotavirus vaccines, Rotarix(®) and RotaTeq(®), are highly efficacious in the developed countries. However, the efficacy of such vaccines in resource deprived countries in Africa and Southeast Asia is low. We reported previously that a bacterially-expressed rotavirus P2-P[8] ΔVP8* subunit vaccine candidate administered intramuscularly elicited high-titers of neutralizing antibodies in guinea pigs and mice and significantly shortened the duration of diarrhea in neonatal gnotobiotic pigs upon oral challenge with virulent human rotavirus Wa strain. To further improve its vaccine potential and provide wider coverage against rotavirus strains of global and regional epidemiologic importance, we constructed 2 tandem recombinant VP8* proteins, P2-P[8] ΔVP8*-P[8] ΔVP8* and P2-P[8] ΔVP8*-P[6] ΔVP8* based on Escherichia coli expression system. The two resulting recombinant tandem proteins were highly soluble and P2-P[8] ΔVP8*-P[8] ΔVP8* was generated with high yield. Moreover, guinea pigs immunized intramuscularly by 3 doses of the P2-P[8] ΔVP8*-P[8] ΔVP8* or P2-P[8] ΔVP8*-P[6] ΔVP8* vaccine with aluminum phosphate adjuvant developed high titers of homotypic and heterotypic neutralizing antibodies against human rotaviruses bearing G1-G4, G8, G9 and G12 with P[8], P[4] or P[6] combination. The results suggest that these 2 subunit vaccines in monovalent or bivalent formulation can provide antigenic coverage to almost all the rotavirus G (VP7) types and major P (VP4) types of global as well as regional epidemiologic importance.

Single-pipetting microfluidic assay device for rapid detection of Salmonella from poultry package.

PubMed

Fronczek, Christopher F; You, David J; Yoon, Jeong-Yeol

2013-02-15

A direct, sensitive, near-real-time, handheld optical immunoassay device was developed to detect Salmonella typhimurium in the naturally occurring liquid from fresh poultry packages (hereafter "chicken matrix"), with just single pipetting of sample (i.e., no filtration, culturing and/or isolation, thus reducing the assay time and the error associated with them). Carboxylated, polystyrene microparticles were covalently conjugated with anti-Salmonella, and the immunoagglutination due to the presence of Salmonella was detected by reading the Mie scatter signals from the microfluidic channels using a handheld device. The presence of chicken matrix did not affect the light scatter signal, since the optical parameters (particle size d, wavelength of incident light λ and scatter angle θ) were optimized to minimize the effect of sample matrix (animal tissues and blood proteins, etc.). The sample was loaded into a microfluidic chip that was split into two channels, one pre-loaded with vacuum-dried, antibody-conjugated particles and the other with vacuum-dried, bovine serum albumin-conjugated particles. This eliminated the need for a separate negative control, effectively minimizing chip-to-chip and sample-to-sample variations. Particles and the sample were diffused in-channel through chemical agitation by Tween 80, also vacuum-dried within the microchannels. Sequential mixing of the sample to the reagents under a strict laminar flow condition synergistically improved the reproducibility and linearity of the assay. In addition, dried particles were shown to successfully detect lower Salmonella concentrations for up to 8 weeks. The handheld device contains simplified circuitry eliminating unnecessary adjustment stages, providing a stable signal, thus maximizing sensitivity. Total assay time was 10 min, and the detection limit 10 CFU mL(-1) was observed in all matrices, demonstrating the suitability of this device for field assays. Copyright © 2012 Elsevier B.V. All rights reserved.

Low-cost far infrared bolometer camera for automotive use

NASA Astrophysics Data System (ADS)

Vieider, Christian; Wissmar, Stanley; Ericsson, Per; Halldin, Urban; Niklaus, Frank; Stemme, Göran; Källhammer, Jan-Erik; Pettersson, Håkan; Eriksson, Dick; Jakobsen, Henrik; Kvisterøy, Terje; Franks, John; VanNylen, Jan; Vercammen, Hans; VanHulsel, Annick

2007-04-01

A new low-cost long-wavelength infrared bolometer camera system is under development. It is designed for use with an automatic vision algorithm system as a sensor to detect vulnerable road users in traffic. Looking 15 m in front of the vehicle it can in case of an unavoidable impact activate a brake assist system or other deployable protection system. To achieve our cost target below €100 for the sensor system we evaluate the required performance and can reduce the sensitivity to 150 mK and pixel resolution to 80 x 30. We address all the main cost drivers as sensor size and production yield along with vacuum packaging, optical components and large volume manufacturing technologies. The detector array is based on a new type of high performance thermistor material. Very thin Si/SiGe single crystal multi-layers are grown epitaxially. Due to the resulting valence barriers a high temperature coefficient of resistance is achieved (3.3%/K). Simultaneously, the high quality crystalline material provides very low 1/f-noise characteristics and uniform material properties. The thermistor material is transferred from the original substrate wafer to the read-out circuit using adhesive wafer bonding and subsequent thinning. Bolometer arrays can then be fabricated using industry standard MEMS process and materials. The inherently good detector performance allows us to reduce the vacuum requirement and we can implement wafer level vacuum packaging technology used in established automotive sensor fabrication. The optical design is reduced to a single lens camera. We develop a low cost molding process using a novel chalcogenide glass (GASIR®3) and integrate anti-reflective and anti-erosion properties using diamond like carbon coating.

Molecular basis for ebolavirus VP35 suppression of human dendritic cell maturation.

PubMed

Yen, Benjamin; Mulder, Lubbertus C F; Martinez, Osvaldo; Basler, Christopher F

2014-11-01

Zaire ebolavirus (EBOV) VP35 is a double-stranded RNA (dsRNA)-binding protein that inhibits RIG-I signaling and alpha/beta interferon (IFN-α/β) responses by both dsRNA-binding-dependent and -independent mechanisms. VP35 also suppresses dendritic cell (DC) maturation. Here, we define the pathways and mechanisms through which VP35 impairs DC maturation. Wild-type VP35 (VP35-WT) and two well-characterized VP35 mutants (F239A and R322A) that independently ablate dsRNA binding and RIG-I inhibition were delivered to primary human monocyte-derived DCs (MDDCs) using a lentivirus-based expression system. VP35-WT suppressed not only IFN-α/β but also proinflammatory responses following stimulation of MDDCs with activators of RIG-I-like receptor (RLR) signaling, including RIG-I activators such as Sendai virus (SeV) or 5'-triphosphate RNA, or MDA5 activators such as encephalomyocarditis virus (EMCV) or poly(I · C). The F239A and R322A mutants exhibited greatly reduced suppression of IFN-α/β and proinflammatory cytokine production following treatment of DCs with RLR agonists. VP35-WT also blocked the upregulation of DC maturation markers and the stimulation of allogeneic T cell responses upon SeV infection, whereas the mutants did not. In contrast to the RLR activators, VP35-WT and the VP35 mutants impaired IFN-β production induced by Toll-like receptor 3 (TLR3) or TLR4 agonists but failed to inhibit proinflammatory cytokine production induced by TLR2, TLR3, or TLR4 agonists. Furthermore, VP35 did not prevent lipopolysaccharide (LPS)-induced upregulation of surface markers of MDDC maturation and did not prevent LPS-triggered allogeneic T cell stimulation. Therefore, VP35 is a general antagonist of DC responses to RLR activation. However, TLR agonists can circumvent many of the inhibitory effects of VP35. Therefore, it may be possible to counteract EBOV immune evasion by using treatments that bypass the VP35-imposed block to DC maturation. The VP35 protein, which is an inhibitor of RIG-I signaling and alpha/beta interferon (IFN-α/β) responses, has been implicated as an EBOV-encoded factor that contributes to suppression of dendritic cell (DC) function. We used wild-type VP35 and previously characterized VP35 mutants to clarify VP35-DC interactions. Our data demonstrate that VP35 is a general inhibitor of RIG-I-like receptor (RLR) signaling that blocks not only RIG-I- but also MDA5-mediated induction of IFN-α/β responses. Furthermore, in DCs, VP35 also impairs the RLR-mediated induction of proinflammatory cytokine production, upregulation of costimulatory markers, and activation of T cells. These inhibitory activities require VP35 dsRNA-binding activity, an activity previously correlated to VP35 RIG-I inhibitory function. In contrast, while VP35 can inhibit IFN-α/β production induced by TLR3 or TLR4 agonists, this occurs in a dsRNA-independent fashion, and VP35 does not inhibit TLR-mediated expression of proinflammatory cytokines. These data suggest strategies to overcome VP35 inhibition of DC function. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Structures of Rotavirus Reassortants Demonstrate Correlation of Altered Conformation of the VP4 Spike and Expression of Unexpected VP4-Associated Phenotypes

PubMed Central

Pesavento, Joseph B.; Billingsley, Angela M.; Roberts, Ed J.; Ramig, Robert F.; Prasad, B. V. Venkataram

2003-01-01

Numerous prior studies have indicated that viable rotavirus reassortants containing structural proteins of heterologous parental origin may express unexpected phenotypes, such as changes in infectivity and immunogenicity. To provide a structural basis for alterations in phenotypic expression, a three-dimensional structural analysis of these reassortants was conducted. The structures of the reassortants show that while VP4 generally maintains the parental structure when moved to a heterologous protein background, in certain reassortants, there are subtle alterations in the conformation of VP4. The alterations in VP4 conformation correlated with expression of unexpected VP4-associated phenotypes. Interactions between heterologous VP4 and VP7 in reassortants expressing unexpected phenotypes appeared to induce the conformational alterations seen in VP4. PMID:12584352

The BepiColombo/SERENA package: Serena Integrated Test campaign

NASA Astrophysics Data System (ADS)

Orsini, S.; De Angelis, E.; Livi, S.; Lichtenegger, H.; Barabash, S.; Milillo, A.; Wurz, P.; Olivieir, A.; D'Arcio, L.; Philips, M.; Laky, G.; Wieser, M.; Camozzi, F.; Di Lellis, A. M.; Rispoli, R.; Jeszenesky, H.; Mura, A.; Aronica, A.; Lazzarotto, F.; Vertolli, N.

2017-09-01

The activities related to the BepiColombo/ MPO/SERENA Integrated Test (SIT, held in February-March 2017 inside the thermal vacuum facility at the University of Bern, Phys. Inst.) are presented. This campaign has been a unique opportunity to test the experiment performances, with all the four flight-spare instruments of SERENA (ELENA, STROFIO, PICAM, and MIPA, simultaneously operated by the System Control Unit (SCU), in a fully operational configuration.

Digital model of a vacuum circuit breaker for the analysis of switching waveforms in electrical circuits

NASA Astrophysics Data System (ADS)

Budzisz, Joanna; Wróblewski, Zbigniew

2016-03-01

The article presents a method of modelling a vaccum circuit breaker in the ATP/EMTP package, the results of the verification of the correctness of the developed digital circuit breaker model operation and its practical usefulness for analysis of overvoltages and overcurrents occurring in commutated capacitive electrical circuits and also examples of digital simulations of overvoltages and overcurrents in selected electrical circuits.

Thematic mapper flight model preshipment review data package. Volume 3, part B: System data

NASA Technical Reports Server (NTRS)

1982-01-01

Procedures and results are presented for performance and systems integration tests of flight model-1 thematic mapper. Aspects considered cover electronic module integration, radiometric calibration, spectral matching, spatial coverage, radiometric calibration of the calibrator, coherent noise, dynamic square wave response, band to band registration, geometric accuracy, and self induced vibration. Thermal vacuum tests, EMI/EMS, and mass properties are included. Liens are summarized.

Spool piece aperture: warm to cold temperature position changes

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

McInturff, A.D.

1982-12-01

In an effort to determine the position of the correction coil system contained in the spool piece in operation, a series of experiments were done in Lab 2 during the cryogenic testing of the spools. A special turn-around box was constructed which had a window through which a sighting could be made of the aperture of the coil packages. A set of four survey monuments were constructed with three point suspension, which were back-lighted by placing a small bulb behind each target which could be turned on individually external to the spool string while under vacuum and at any temperature.more » The targets were located at the ends of each correction coil package in their beam tube.« less

Refined liquid smoke: a potential antilisterial additive to cold-smoked sockeye salmon (Oncorhynchus nerka).

PubMed

Montazeri, Naim; Himelbloom, Brian H; Oliveira, Alexandra C M; Leigh, Mary Beth; Crapo, Charles A

2013-05-01

Cold-smoked salmon (CSS) is a potentially hazardous ready-to-eat food product due to the high risk of contamination with Listeria monocytogenes and lack of a listericidal step. We investigated the antilisterial property of liquid smokes (LS) against Listeria innocua ATCC 33090 (surrogate to L. monocytogenes) as a potential supplement to vacuum-packaged CSS. A full-strength LS (Code 10-Poly), and three commercially refined fractions (AM-3, AM-10, and 1291) having less color and flavor (lower content of phenols and carbonyl-containing compounds) were tested. In vitro assays showed strong inhibition for all LS except for 1291. The CSS strips were surface coated with AM-3 and AM-10 at 1% LS (vol/wt) with an L-shaped glass rod and then inoculated with L. innocua at 3.5 log CFU/g, vacuum packaged, and stored at 4°C. The LS did not completely eliminate L. innocua but provided a 2-log reduction by day 14, with no growth up to 35 days of refrigerated storage. A simple difference sensory test by 180 untrained panelists showed the application of AM-3 did not significantly influence the overall sensorial quality of CSS. In essence, the application of the refined LS as an antilisterial additive to CSS is recommended.

Ubiquitin Ligase WWP1 Interacts with Ebola Virus VP40 To Regulate Egress.

PubMed

Han, Ziying; Sagum, Cari A; Takizawa, Fumio; Ruthel, Gordon; Berry, Corbett T; Kong, Jing; Sunyer, J Oriol; Freedman, Bruce D; Bedford, Mark T; Sidhu, Sachdev S; Sudol, Marius; Harty, Ronald N

2017-10-15

Ebola virus (EBOV) is a member of the Filoviridae family and the cause of hemorrhagic fever outbreaks. The EBOV VP40 (eVP40) matrix protein is the main driving force for virion assembly and budding. Indeed, expression of eVP40 alone in mammalian cells results in the formation and budding of virus-like particles (VLPs) which mimic the budding process and morphology of authentic, infectious EBOV. To complete the budding process, eVP40 utilizes its PPXY L-domain motif to recruit a specific subset of host proteins containing one or more modular WW domains that then function to facilitate efficient production and release of eVP40 VLPs. In this report, we identified additional host WW-domain interactors by screening for potential interactions between mammalian proteins possessing one or more WW domains and WT or PPXY mutant peptides of eVP40. We identified the HECT family E3 ubiquitin ligase WWP1 and all four of its WW domains as strong interactors with the PPXY motif of eVP40. The eVP40-WWP1 interaction was confirmed by both peptide pulldown and coimmunoprecipitation assays, which also demonstrated that modular WW domain 1 of WWP1 was most critical for binding to eVP40. Importantly, the eVP40-WWP1 interaction was found to be biologically relevant for VLP budding since (i) small interfering RNA (siRNA) knockdown of endogenous WWP1 resulted in inhibition of eVP40 VLP egress, (ii) coexpression of WWP1 and eVP40 resulted in ubiquitination of eVP40 and a subsequent increase in eVP40 VLP egress, and (iii) an enzymatically inactive mutant of WWP1 (C890A) did not ubiquitinate eVP40 or enhance eVP40 VLP egress. Last, our data show that ubiquitination of eVP40 by WWP1 enhances egress of VLPs and concomitantly decreases cellular levels of higher-molecular-weight oligomers of eVP40. In sum, these findings contribute to our fundamental understanding of the functional interplay between host E3 ligases, ubiquitination, and regulation of EBOV VP40-mediated egress. IMPORTANCE Ebola virus (EBOV) is a high-priority, emerging human pathogen that can cause severe outbreaks of hemorrhagic fever with high mortality rates. As there are currently no approved vaccines or treatments for EBOV, a better understanding of the biology and functions of EBOV-host interactions that promote or inhibit viral budding is warranted. Here, we describe a physical and functional interaction between EBOV VP40 (eVP40) and WWP1, a host E3 ubiquitin ligase that ubiquitinates VP40 and regulates VLP egress. This viral PPXY-host WW domain-mediated interaction represents a potential new target for host-oriented inhibitors of EBOV egress. Copyright © 2017 American Society for Microbiology.

Expression and responses to dehydration and salinity stresses of V-PPase gene members in wheat.

PubMed

Wang, Yuezhi; Xu, Haibin; Zhang, Guangxiang; Zhu, Huilan; Zhang, Lixia; Zhang, Zhengzhi; Zhang, Caiqin; Ma, Zhengqiang

2009-12-01

Vacuolar H(+)-translocating pyrophosphatase (V-PPase) is a key enzyme related to plant growth as well as abiotic stress tolerance. In this work, wheat V-PPase genes TaVP1, TaVP2 and TaVP3 were identified. TaVP1 and TaVP2 are more similar to each other than to TaVP3. Their deduced polypeptide sequences preserve the topological structure and essential residues of V-PPases. Phylogenetic studies suggested that monocot plants, at least monocot grasses, have three VP paralogs. TaVP3 transcripts were only detected in developing seeds, and no TaVP2 transcripts were found in germinating seeds. TaVP2 was mainly expressed in shoot tissues and down-regulated in leaves under dehydration. Its expression was up-regulated in roots under high salinity. TaVP1 was relatively more ubiquitously and evenly expressed than TaVP2. Its expression level in roots was highest among the tissues examined, and was inducible by salinity stress. These results indicated that the V-PPase gene paralogs in wheat are differentially regulated spatially and in response to dehydration and salinity stresses. 2009 Institute of Genetics and Developmental Biology and the Genetics Society of China. Published by Elsevier Ltd. All rights reserved.

Spontaneous Evolution of Nanostructure in Composite Films Consisting of Mixtures of Two Different Block Copolymer Micelles

NASA Astrophysics Data System (ADS)

Kim, Sehee; Char, Kookheon; Sohn, Byeong-Hyeok

2010-03-01

Diblock copolymers consisting of two immiscible polymer blocks covalently bonded together form various self-assembled nanostructures such as spheres, cylinders, and lamellae in bulk phase. In a selective solvent, however, they assemble into micelles with soluble corona brushes and immiscible cores. Both polystyrene-poly(4-vinylpyridine) (PS-b-P4VP) and polystyrene-poly(2-vinylpyridine) (PS-b-P2VP) diblock copolymers form micelles with PS coronas and P4VP or P2VP cores in a PS selective solvent (toluene). By varying the mixture ratio between PS-b-P4VP and PS-b-P2VP, composite films based on the micellar mixtures of PS-b-P4VP and PS-b-P2VP were obtained by spin-coating, followed by the solvent annealing with tetrahydrofuran (THF) vapor. Since THF is a solvent for both PS and P2VP blocks and, at the same time, a non-solvent for the P4VP block, PS-P2VP micelles transformed to lamellar multilayers while PS-P4VP micelles remained intact during the THF annealing. The spontaneous evolution of nanostructure in composite films consisting of lamellae layers with BCP micelles were investigated in detail by cross-sectional TEM and AFM.

Cryo-Electron Microscopy Reconstruction Shows Poliovirus 135S Particles Poised for Membrane Interaction and RNA Release

PubMed Central

Butan, Carmen; Filman, David J.

2014-01-01

During infection, binding of mature poliovirus to cell surface receptors induces an irreversible expansion of the capsid, to form an infectious cell-entry intermediate particle that sediments at 135S. In these expanded virions, the major capsid proteins (VP1 to VP3) adopt an altered icosahedral arrangement to open holes in the capsid at 2-fold and quasi-3-fold axes, and internal polypeptides VP4 and the N terminus of VP1, which can bind membranes, become externalized. Cryo-electron microscopy images for 117,330 particles were collected using Leginon and reconstructed using FREALIGN. Improved rigid-body positioning of major capsid proteins established reliably which polypeptide segments become disordered or rearranged. The virus-to-135S transition includes expansion of 4%, rearrangements of the GH loops of VP3 and VP1, and disordering of C-terminal extensions of VP1 and VP2. The N terminus of VP1 rearranges to become externalized near its quasi-3-fold exit, binds to rearranged GH loops of VP3 and VP1, and attaches to the top surface of VP2. These details improve our understanding of subsequent stages of infection, including endocytosis and RNA transfer into the cytoplasm. PMID:24257617

Maturation of the Hepatitis A Virus Capsid Protein VP1 Is Not Dependent on Processing by the 3Cpro Proteinase

PubMed Central

Martin, Annette; Bénichou, Danièle; Chao, Shih-Fong; Cohen, Lisette M.; Lemon, Stanley M.

1999-01-01

Most details of the processing of the hepatitis A virus (HAV) polyprotein are known. Unique among members of the family Picornaviridae, the primary cleavage of the HAV polyprotein is mediated by 3Cpro, the only proteinase known to be encoded by the virus, at the 2A/2B junction. All other cleavages of the polyprotein have been considered to be due to 3Cpro, although the precise location and mechanism responsible for the VP1/2A cleavage have been controversial. Here we present data that argue strongly against the involvement of the HAV 3Cpro proteinase in the maturation of VP1 from its VP1-2A precursor. Using a heterologous expression system based on recombinant vaccinia viruses directing the expression of full-length or truncated capsid protein precursors, we show that the C terminus of the mature VP1 capsid protein is located near residue 764 of the polyprotein. However, a proteolytically active HAV 3Cpro that was capable of directing both VP0/VP3 and VP3/VP1 cleavages in vaccinia virus-infected cells failed to process the VP1-2A precursor. Using site-directed mutagenesis of an infectious molecular clone of HAV, we modified potential VP1/2A cleavage sites that fit known 3Cpro recognition criteria and found that a substitution that ablates the presumed 3Cpro dipeptide recognition sequence at Glu764-Ser765 abolished neither infectivity nor normal VP1 maturation. Altered electrophoretic mobility of VP1 from a viable mutant virus with an Arg764 substitution indicated that this residue is present in VP1 and that the VP1/2A cleavage occurs downstream of this residue. These data indicate that maturation of the HAV VP1 capsid protein is not dependent on 3Cpro processing and may thus be uniquely dependent on a cellular proteinase. PMID:10400711

The effect of adopting new storage methods for extending product validity periods on manufacturers expected inventory costs.

PubMed

Chen, Po-Yu

2014-01-01

The validness of the expiration dates (validity period) that manufacturers provide on food product labels is a crucial food safety problem. Governments must study how to use their authority by implementing fair awards and punishments to prompt manufacturers into adopting rigorous considerations, such as the effect of adopting new storage methods for extending product validity periods on expected costs. Assuming that a manufacturer sells fresh food or drugs, this manufacturer must respond to current stochastic demands at each unit of time to determine the purchase amount of products for sale. If this decision maker is capable and an opportunity arises, new packaging methods (e.g., aluminum foil packaging, vacuum packaging, high-temperature sterilization after glass packaging, or packaging with various degrees of dryness) or storage methods (i.e., adding desiccants or various antioxidants) can be chosen to extend the validity periods of products. To minimize expected costs, this decision maker must be aware of the processing costs of new storage methods, inventory standards, inventory cycle lengths, and changes in relationships between factors such as stochastic demand functions in a cycle. Based on these changes in relationships, this study established a mathematical model as a basis for discussing the aforementioned topics.

Interplay Among Constitutes of Ebola Virus: Nucleoprotein, Polymerase L, Viral Proteins

NASA Astrophysics Data System (ADS)

Zhang, Minchuan; He, Peiming; Su, Jing; Singh, Dadabhai T.; Su, Hailei; Su, Haibin

Ebola virus is a highly lethal filovirus, claimed thousands of people in its recent outbreak. Seven viral proteins constitute ebola viral structure, and four of them (nucleoprotein (NP), polymerase L, VP35 and VP30) participate majorly in viral replication and transcription. We have elucidated a conformation change of NP cleft by VP35 NP-binding protein domains through superimposing two experimental NP structure images and discussed the function of this conformation change in the replication and transcription with polymerase complex (L, VP35 and VP30). The important roles of VP30 in viral RNA synthesis have also been discussed. A “tapping” model has been proposed in this paper for a better understanding of the interplay among the four viral proteins (NP, polymerase L, VP35 and VP30). Moreover, we have pinpointed some key residue changes on NP (both NP N- and C-terminal) and L between Reston and Zaire by computational studies. Together, this paper provides a description of interactions among ebola viral proteins (NP, L, VP35, VP30 and VP40) in viral replication and transcription, and sheds light on the complex system of viral reproduction.

[The relevance of junctional rhythm during neurocardiogenic reaction provoked by tilt testing].

PubMed

Zyśko, Dorota; Gajek, Jacek; Agrawal, Anil Kumar; Rudnicki, Jerzy

2012-01-01

During neurocardiogenic reaction provoked by tilt testing (TT), different arrhythmias such as sinus bradycardia, sinus arrest, atrioventricular block or junctional rhythm or beats (JR) may occur. The characteristics of the JR during neurocardiogenic reaction have not yet been systematically assessed. It is not known whether the presence of JR during neurocardiogenic reaction is related to clinical characteristics of syncopal patients or the outcome of TT. To assess whether clinical outcome of TT and clinical data are related to the presence of JR during TT. The study group consisted of 532 patients aged 43.3 ± 18.2 years with positive TT, divided into four groups on the basis of the presence of JR and/or a ventricular pause (VP) during neurocardiogenic reaction: group VP(-)/JR(+) - JR present and VP absent, group VP(+)/JR(+) - both JR and VP present, group VP(+)/JR(-) - JR absent and VP present, and group VP(-)/JR(-) - both JR and VP absent. The control group consisted of 53 patients with no history of syncope or presyncope, including 46 patients with negative TT and seven patients with false positive TT. Total loss of consciousness during TT occurred in group VP(-)/JR(+) less frequently than in groups VP(+)/JR(+) and VP(+)/JR(-), and more frequently than in group VP(-)/JR(-) (80% vs 96% vs 94% vs 62%; p 〈 0.05 for both comparisons). Group VP(-)/JR(+) was significantly younger than group VP(-)/JR(-) (37.3 ± 16.3 years vs 45.8 ± 18.9 years; p 〈 0.05) and had a lower number of syncopal events than group VP(+)/JR(+) and VP(+)/JR(-) (median [IQ]: 2.5 (1-6) vs 4 (2-12) and 4 (2-10), respectively; p 〈 0.05) and lower rate of traumatic injuries than group VP(+)/JR(+) and VP(+)/JR(-) (22% vs 45% and 39%, respectively; p 〈 0.05). Logistic regression analysis revealed that the presence of JR was associated with younger age, male gender, history of blood-instrumentation-injection phobia and higher number of syncopal spells in medical history. The ROC curve analysis revealed that a junctional rate of no more than 49 bpm was related to the total loss of consciousness during TT (p 〈 0.05). 1. JR frequently occurs during positive TT and in no subjects with negative TT. 2. Among patients with JR, two groups may be chosen on the basis of a VP occurrence, and these groups differ in respect to clinical characteristics and TT outcome. 3. Relatively rapid JR without VP is related to consciousness preservation during neurocardiogenic reaction at TT and fewer syncopal spells as well as syncope associated with injury in the past. 4. In patients with JR and VP, the JR is slower, of shorter duration, and more frequently single or pairs of junctional beats occur, which indicates high parasympathetic activity, whereas relatively rapid and stable JR may be the symptom of simultaneously increased sympathetic and parasympathetic activity.

Trypsin activation pathway of rotavirus infectivity.

PubMed Central

Arias, C F; Romero, P; Alvarez, V; López, S

1996-01-01

The infectivity of rotaviruses is increased by and most probably is dependent on trypsin treatment of the virus. This proteolytic treatment specifically cleaves VP4, the protein that forms the spikes on the surface of the virions, to polypeptides VP5 and VP8. This cleavage has been reported to occur in rotavirus SA114fM at two conserved, closely spaced arginine residues located at VP4 amino acids 241 and 247. In this work, we have characterized the VP4 cleavage products of rotavirus SA114S generated by in vitro treatment of the virus with increasing concentrations of trypsin and with proteases AspN and alpha-chymotrypsin. The VP8 and VP5 polypeptides were analyzed by gel electrophoresis and by Western blotting (immunoblotting) with antibodies raised to synthetic peptides that mimic the terminal regions of VP4 generated by the trypsin cleavage. It was shown that in addition to arginine residues 241 and 247, VP4 is cleaved at arginine residue 231. These three sites were found to have different susceptibilities to trypsin, Arg-241 > Arg-231 > Arg-247, with the enhancement of infectivity correlating with cleavage at Arg-247 rather than at Arg-231 or Arg-241. Proteases AspN and alpha-chymotrypsin cleaved VP4 at Asp-242 and Tyr-246, respectively, with no significant enhancement of infectivity, although this enhancement could be achieved by further treatment of the virus with trypsin. The VP4 end products of trypsin treatment were a homogeneous VP8 polypeptide comprising VP4 amino acids 1 to 231 and a heterogeneous VP5, which is formed by two polypeptide species (present at a ratio of approximately 1:5) as a result of cleavage at either Arg-241 or Arg-247. A pathway for the trypsin activation of rotavirus infectivity is proposed. PMID:8709201

Image quality and radiation dose of brain computed tomography in children: effects of decreasing tube voltage from 120 kVp to 80 kVp.

PubMed

Park, Ji Eun; Choi, Young Hun; Cheon, Jung-Eun; Kim, Woo Sun; Kim, In-One; Cho, Hyun Suk; Ryu, Young Jin; Kim, Yu Jin

2017-05-01

Computed tomography (CT) has generated public concern associated with radiation exposure, especially for children. Lowering the tube voltage is one strategy to reduce radiation dose. To assess the image quality and radiation dose of non-enhanced brain CT scans acquired at 80 kilo-voltage peak (kVp) compared to those at 120 kVp in children. Thirty children who had undergone both 80- and 120-kVp non-enhanced brain CT were enrolled. For quantitative analysis, the mean attenuation of white and gray matter, attenuation difference, noise, signal-to-noise ratio, contrast-to-noise ratio and posterior fossa artifact index were measured. For qualitative analysis, noise, gray-white matter differentiation, artifact and overall image quality were scored. Radiation doses were evaluated by CT dose index, dose-length product and effective dose. The mean attenuations of gray and white matter and contrast-to-noise ratio were significantly increased at 80 kVp, while parameters related to image noise, i.e. noise, signal-to-noise ratio and posterior fossa artifact index were higher at 80 kVp than at 120 kVp. In qualitative analysis, 80-kVp images showed improved gray-white differentiation but more artifacts compared to 120-kVp images. Subjective image noise and overall image quality scores were similar between the two scans. Radiation dose parameters were significantly lower at 80 kVp than at 120 kVp. In pediatric non-enhanced brain CT scans, a decrease in tube voltage from 120 kVp to 80 kVp resulted in improved gray-white matter contrast, comparable image quality and decreased radiation dose.

Recombination and Population Mosaic of a Multifunctional Viral Gene, Adeno-Associated Virus cap

PubMed Central

Takeuchi, Yasuhiro; Myers, Richard; Danos, Olivier

2008-01-01

Homologous recombination is a dominant force in evolution and results in genetic mosaics. To detect evidence of recombination events and assess the biological significance of genetic mosaics, genome sequences for various viral populations of reasonably large size are now available in the GenBank. We studied a multi-functional viral gene, the adeno-associated virus (AAV) cap gene, which codes for three capsid proteins, VP1, VP2 and VP3. VP1-3 share a common C-terminal domain corresponding to VP3, which forms the viral core structure, while the VP1 unique N-terminal part contains an enzymatic domain with phospholipase A2 activity. Our recombinant detection program (RecI) revealed five novel recombination events, four of which have their cross-over points in the N-terminal, VP1 and VP2 unique region. Comparison of phylogenetic trees for different cap gene regions confirmed discordant phylogenies for the recombinant sequences. Furthermore, differences in the phylogenetic tree structures for the VP1 unique (VP1u) region and the rest of cap highlighted the mosaic nature of cap gene in the AAV population: two dominant forms of VP1u sequences were identified and these forms are linked to diverse sequences in the rest of cap gene. This observation together with the finding of frequent recombination in the VP1 and 2 unique regions suggests that this region is a recombination hot spot. Recombination events in this region preserve protein blocks of distinctive functions and contribute to convergence in VP1u and divergence of the rest of cap. Additionally the possible biological significance of two dominant VP1u forms is inferred. PMID:18286191

Value of 100 kVp scan with sinogram-affirmed iterative reconstruction algorithm on a single-source CT system during whole-body CT for radiation and contrast medium dose reduction: an intra-individual feasibility study.

PubMed

Nagayama, Y; Nakaura, T; Oda, S; Tsuji, A; Urata, J; Furusawa, M; Tanoue, S; Utsunomiya, D; Yamashita, Y

2018-02-01

To perform an intra-individual investigation of the usefulness of a contrast medium (CM) and radiation dose-reduction protocol using single-source computed tomography (CT) combined with 100 kVp and sinogram-affirmed iterative reconstruction (SAFIRE) for whole-body CT (WBCT; chest-abdomen-pelvis CT) in oncology patients. Forty-three oncology patients who had undergone WBCT under both 120 and 100 kVp protocols at different time points (mean interscan intervals: 98 days) were included retrospectively. The CM doses for the 120 and 100 kVp protocols were 600 and 480 mg iodine/kg, respectively; 120 kVp images were reconstructed with filtered back-projection (FBP), whereas 100 kVp images were reconstructed with FBP (100 kVp-F) and the SAFIRE (100 kVp-S). The size-specific dose estimate (SSDE), iodine load and image quality of each protocol were compared. The SSDE and iodine load of 100 kVp protocol were 34% and 21%, respectively, lower than of 120 kVp protocol (SSDE: 10.6±1.1 versus 16.1±1.8 mGy; iodine load: 24.8±4versus 31.5±5.5 g iodine, p<0.01). Contrast enhancement, objective image noise, contrast-to-noise-ratio, and visual score of 100 kVp-S were similar to or better than of 120 kVp protocol. Compared with the 120 kVp protocol, the combined use of 100 kVp and SAFIRE in WBCT for oncology assessment with an SSCT facilitated substantial reduction in the CM and radiation dose while maintaining image quality. Copyright © 2017 The Royal College of Radiologists. Published by Elsevier Ltd. All rights reserved.

Investigating Ebola virus pathogenicity using molecular dynamics.

PubMed

Pappalardo, Morena; Collu, Francesca; Macpherson, James; Michaelis, Martin; Fraternali, Franca; Wass, Mark N

2017-08-11

Ebolaviruses have been known to cause deadly disease in humans for 40 years and have recently been demonstrated in West Africa to be able to cause large outbreaks. Four Ebolavirus species cause severe disease associated with high mortality in humans. Reston viruses are the only Ebolaviruses that do not cause disease in humans. Conserved amino acid changes in the Reston virus protein VP24 compared to VP24 of other Ebolaviruses have been suggested to alter VP24 binding to host cell karyopherins resulting in impaired inhibition of interferon signalling, which may explain the difference in human pathogenicity. Here we used protein structural analysis and molecular dynamics to further elucidate the interaction between VP24 and KPNA5. As a control experiment, we compared the interaction of wild-type and R137A-mutant (known to affect KPNA5 binding) Ebola virus VP24 with KPNA5. Results confirmed that the R137A mutation weakens direct VP24-KPNA5 binding and enables water molecules to penetrate at the interface. Similarly, Reston virus VP24 displayed a weaker interaction with KPNA5 than Ebola virus VP24, which is likely to reduce the ability of Reston virus VP24 to prevent host cell interferon signalling. Our results provide novel molecular detail on the interaction of Reston virus VP24 and Ebola virus VP24 with human KPNA5. The results indicate a weaker interaction of Reston virus VP24 with KPNA5 than Ebola virus VP24, which is probably associated with a decreased ability to interfere with the host cell interferon response. Hence, our study provides further evidence that VP24 is a key player in determining Ebolavirus pathogenicity.

Characterization of the DNA binding properties of polyomavirus capsid protein

NASA Technical Reports Server (NTRS)

Chang, D.; Cai, X.; Consigli, R. A.; Spooner, B. S. (Principal Investigator)

1993-01-01

The DNA binding properties of the polyomavirus structural proteins VP1, VP2, and VP3 were studied by Southwestern analysis. The major viral structural protein VP1 and host-contributed histone proteins of polyomavirus virions were shown to exhibit DNA binding activity, but the minor capsid proteins VP2 and VP3 failed to bind DNA. The N-terminal first five amino acids (Ala-1 to Lys-5) were identified as the VP1 DNA binding domain by genetic and biochemical approaches. Wild-type VP1 expressed in Escherichia coli (RK1448) exhibited DNA binding activity, but the N-terminal truncated VP1 mutants (lacking Ala-1 to Lys-5 and Ala-1 to Cys-11) failed to bind DNA. The synthetic peptide (Ala-1 to Cys-11) was also shown to have an affinity for DNA binding. Site-directed mutagenesis of the VP1 gene showed that the point mutations at Pro-2, Lys-3, and Arg-4 on the VP1 molecule did not affect DNA binding properties but that the point mutation at Lys-5 drastically reduced DNA binding affinity. The N-terminal (Ala-1 to Lys-5) region of VP1 was found to be essential and specific for DNA binding, while the DNA appears to be non-sequence specific. The DNA binding domain and the nuclear localization signal are located in the same N-terminal region.

Two-dimensional seismic image of the San Andreas Fault in the Northern Gabilan Range, central California: Evidence for fluids in the fault zone

USGS Publications Warehouse

Thurber, C.; Roecker, S.; Ellsworth, W.; Chen, Y.; Lutter, W.; Sessions, R.

1997-01-01

A joint inversion for two-dimensional P-wave velocity (Vp), P-to-S velocity ratio (Vp/Vs), and earthquake locations along the San Andreas fault (SAF) in central California reveals a complex relationship among seismicity, fault zone structure, and the surface fault trace. A zone of low Vp and high Vp/Vs lies beneath the SAF surface trace (SAFST), extending to a depth of about 6 km. Most of the seismic activity along the SAF occurs at depths of 3 to 7 km in a southwest-dipping zone that roughly intersects the SAFST, and lies near the southwest edge of the low Vp and high Vp/Vs zones. Tests indicate that models in which this seismic zone is significantly closer to vertical can be confidently rejected. A second high Vp/Vs zone extends to the northeast, apparently dipping beneath the Diablo Range. Another zone of seismicity underlies the northeast portion of this Vp/Vs high. The high Vp/Vs zones cut across areas of very different Vp values, indicating that the high Vp/Vs values are due to the presence of fluids, not just lithology. The close association between the zones of high Vp/Vs and seismicity suggests a direct involvement of fluids in the faulting process. Copyright 1997 by the American Geophysical Union.

Consumers' use of written product information.

PubMed

Wiese, Bettina S; Sauer, Jürgen; Rüttinger, Bruno

2004-09-15

Two studies were conducted to investigate the predictive role of person-specific, product-specific, and situation-specific influences on the use of instruction manuals in the field of electrical consumer products. In a laboratory study, 42 participants were observed while putting a vacuum cleaner into operation. Situational primes (i.e., receiving a verbal cue that the packaging contains an instruction manual) increased the probability of the user manual being read. Additional verbal information that the manual contains information on energy-saving behaviours was especially motivating for persons with high environmental concern. Self-report data, collected on a wide range of products, suggest that product complexity is the best predictor of instruction manual use. In a second study with 30 participants, different positions of product labels were compared, i.e. placing the information on the packaging or directly onto the product. Information placed directly onto the product had a significantly higher influence on participants' actual behaviour than providing the same information on the packaging.

Effects of soy sauce and packaging method on volatile compounds and lipid oxidation of cooked irradiated beef patties

NASA Astrophysics Data System (ADS)

Kim, Hyun-Wook; Lee, Soo-Yeon; Hwang, Ko-Eun; Song, Dong-Heon; Kim, Yong-Jae; Ham, Yun-Kyung; Lee, Choong-Hee; Choi, Yun-Sang; Lee, Ju-Woon; Kim, Cheon-Jei

2014-10-01

The objective of this study is to determine the effect of soy sauce on volatile compounds and lipid oxidation of cooked irradiated beef patties. Sulfur-containing volatile components, which are produced by irradiation, were not found in all treatments. Volatile components derived from soy sauce, such as 3-hydroxy-2-butanone, acetic acid, 3-methyl-1-butanol and 2-methyl-1-butanol, were detected in beef patties containing soy sauce regardless of irradiation and packaging method. Volatile aldehydes, including hexanal, significantly decreased the irradiated beef patty prepared with soy sauce compared to those of irradiated beef patty made with NaCl at 1 day and 5 days after irradiation. In addition, combined use of vacuum packaging and soy sauce treatments could inhibit the formation of volatile compounds and 2-thiobarbituric acid reactive substances during chilled storage. Therefore, the use of soy sauce in cooked and irradiated beef could reduce the production of volatile components associated with the irradiation-induced off-flavor and lipid oxidation.

Opportunities for bio-based packaging technologies to improve the quality and safety of fresh and further processed muscle foods.

PubMed

Cutter, Catherine Nettles

2006-09-01

It has been well documented that vacuum or modified atmosphere packaging materials, made from polyethylene- or other plastic-based materials, have been found to improve the stability and safety of raw or further processed muscle foods. However, recent research developments have demonstrated the feasibility, utilization, and commercial application of a variety of bio-based polymers or bio-polymers made from a variety of materials, including renewable/sustainable agricultural commodities, and applied to muscle foods. A variety of these bio-based materials have been shown to prevent moisture loss, drip, reduce lipid oxidation and improve flavor attributes, as well as enhancing the handling properties, color retention, and microbial stability of foods. With consumers demanding more environmentally friendly packaging and a desire for more natural products, bio-based films or bio-polymers will continue to play an important role in the food industry by improving the quality of many products, including fresh or further processed muscle foods.

Expression, purification, crystallization and preliminary X-ray diffraction analysis of the VP8* sialic acid-binding domain of porcine rotavirus strain OSU

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

Zhang, Yang-De, E-mail: zhangyd1960@yahoo.com.cn; Li, Hao; Liu, Hui

2007-02-01

Porcine rotavirus strain OSU VP8* domain has been expressed, purified and crystallized. X-ray diffraction data from different crystal forms of the VP8* domain have been collected to 2.65 and 2.2 Å resolution, respectively. The rotavirus outer capsid spike protein VP4 is utilized in the process of rotavirus attachment to and membrane penetration of host cells. VP4 is cleaved by trypsin into two domains: VP8* and VP5*. The VP8* domain is implicated in initial interaction with sialic acid-containing cell-surface carbohydrates and triggers subsequent virus invasion. The VP8* domain from porcine OSU rotavirus was cloned and expressed in Escherichia coli. Different crystalmore » forms (orthorhombic P2{sub 1}2{sub 1}2{sub 1} and tetragonal P4{sub 1}2{sub 1}2) were harvested from two distinct crystallization conditions. Diffraction data have been collected to 2.65 and 2.2 Å resolution and the VP8*{sub 65–224} structure was determined by molecular replacement.« less

Antigenicity analysis of human parvovirus B19-VP1u protein in the induction of anti-phospholipid syndrome.

PubMed

Lin, Chun-Yu; Chiu, Chun-Ching; Cheng, Ju; Lin, Chia-Yun; Shi, Ya-Fang; Tsai, Chun-Chou; Tzang, Bor-Show; Hsu, Tsai-Ching

2018-01-01

Mounting evidence suggests a connection between human parvovirus B19 (B19) and autoimmune diseases, and especially an association between the B19-VP1 unique region (VP1u) and anti-phospholipid syndrome (APS). However, little is known about the antigenicity of B19-VP1u in the induction of APS-like syndrome. To elucidate the antigenicity of B19-VP1u in the induction of APS, N-terminal truncated B19-VP1u (tVP1u) proteins were prepared to immunize Balb/c mice to generate antibodies against B19-tVP1u proteins. The secreted phospholipase A2 (sPLA2) activities and binding specificity of mice anti-B19-tVP1u antibodies with cardiolipin (CL) and beta-2-glycoprotein I (β2GPI) were evaluated by performing immunoblot, ELISA and absorption experiments. A mice model of passively induced APS was adopted. Although sPLA2 activities were identified in all B19-tVP1u proteins, only amino acid residues 61-227 B19-tVP1u exhibited a higher sPLA2 activity. Autoantibodies against CL and β2GPI exhibited binding activities with all B19-tVP1u proteins. IgG that was purified from mice that had been immunized with amino acid residues 21-227 to 121-227 B19-tVP1u proteins exhibited significantly higher binding activity with CL. IgG that was purified from mice that had been immunized with amino acid residues 21-227, 31-227, 82-227 and 91-227 B19-tVP1u proteins exhibited significantly higher binding activity with β2GPI. Accordingly, significantly higher binding inhibition of CL was detected in the presence of amino acid residues 61-227 and 101-227 B19-tVP1u. Significantly higher binding inhibition of β2GPI was detected in the presence of amino acid residues 21-227, 31-227, 82-227 and 91-227 B19-tVP1u. The mice that received amino acid residues 31-227 or 61-227 anti-tB19-VP1u IgG revealed significant thrombocytopenia and those that received amino acid residues 21-227, 31-227, 61-227, 71-227, 82-227, 91-227, 101-227 or 114-227 anti-tB19-VP1u IgG exhibited significantly prolonged aPTT. These findings provide further information concerning the role of B19-VP1u antigenicity in APS-like autoimmunity.

Production of biogenic amines by lactic acid bacteria and enterobacteria isolated from fresh pork sausages packaged in different atmospheres and kept under refrigeration.

PubMed

Curiel, J A; Ruiz-Capillas, C; de Las Rivas, B; Carrascosa, A V; Jiménez-Colmenero, F; Muñoz, R

2011-07-01

The occurrence of in vitro amino acid activity in bacterial strains associated with fresh pork sausages packaged in different atmospheres and kept in refrigeration was studied. The presence of biogenic amines in decarboxylase broth was confirmed by ion-exchange chromatography and by the presence of the corresponding decarboxylase genes by PCR. From the 93 lactic acid bacteria and 100 enterobacteria strains analysed, the decarboxylase medium underestimates the number of biogenic amine-producer strains. 28% of the lactic acid bacteria produced tyramine and presented the tdc gene. All the tyramine-producer strains were molecularly identified as Carnobacterium divergens. Differences on the relative abundance of C. divergens were observed among the different packaging atmospheres assayed. After 28 days of storage, the presence of argon seems to inhibit C. divergens growth, while packing under vacuum seems to favour it. Among enterobacteria, putrescine was the amine more frequently produced (87%), followed by cadaverine (85%); agmatine and tyramine were only produced by 13 and 1%, respectively, of the strains analysed. Packing under vacuum or in an atmosphere containing nitrogen seems to inhibit the growth of enterobacteria which produce simultaneously putrescine, cadaverine, and agmatine. Contrarily, over-wrapping or packing in an atmosphere containing argon seems to favour the growth of agmatine producer-enterobacteria. The production of putrescine and cadaverine was associated with the presence of the corresponding amino acid decarboxylase genes. The biogenic amine-producer strains were included in a wide range of enterobacterial species, including Kluyvera intermedia, Enterobacter aerogenes, Yersinia kristensenii, Serratia grimesii, Serratia ficaria, Yersinia rodhei, Providencia vermicola and Obesumbacterium proteus. Copyright © 2011 Elsevier Ltd. All rights reserved.

Purification of recombinant budgerigar fledgling disease virus VP1 capsid protein and its ability for in vitro capsid assembly

NASA Technical Reports Server (NTRS)

Rodgers, R. E.; Chang, D.; Cai, X.; Consigli, R. A.; Spooner, B. S. (Principal Investigator)

1994-01-01

A recombinant system for the major capsid VP1 protein of budgerigar fledgling disease virus has been established. The VP1 gene was inserted into a truncated form of the pFlag-1 vector and expressed in Escherichia coli. The budgerigar fledgling disease virus VP1 protein was purified to near homogeneity by immunoaffinity chromatography. Fractions containing highly purified VP1 were pooled and found to constitute 3.3% of the original E. coli-expressed VP1 protein. Electron microscopy revealed that the VP1 protein was isolated as pentameric capsomeres. Electron microscopy also revealed that capsid-like particles were formed in vitro from purified VP1 capsomeres with the addition of Ca2+ ions and the removal of chelating and reducing agents.

Using Virtual Patients to Teach Empathy: A Randomized Controlled Study to Enhance Medical Students' Empathic Communication.

PubMed

Foster, Adriana; Chaudhary, Neelam; Kim, Thomas; Waller, Jennifer L; Wong, Joyce; Borish, Michael; Cordar, Andrew; Lok, Benjamin; Buckley, Peter F

2016-06-01

Physician empathy is a complex phenomenon known to improve illness outcomes; however, few tools are available for deliberate practice of empathy. We used a virtual patient (VP) to teach empathic communication to first-year medical students. We then evaluated students' verbal empathy in a standardized patient (SP) interaction. Seventy medical students, randomly assigned to 3 separate study groups, interacted with (1) a control VP portraying depression, (2) a VP with a backstory simulating patient shadowing, or (3) a VP able to give immediate feedback about empathic communication (empathy-feedback VP). Subsequently, the students interviewed an SP portraying a scenario that included opportunities to express empathy. All SP interviews were recorded and transcribed. The study outcomes were (1) the students' verbal response to the empathic opportunities presented by the SP, as coded by reliable assessors using the Empathic Communication Coding System, and (2) the students' responses as coded by the SPs, using a communication checklist. There were no significant differences in student demographics between groups. The students who interacted with the empathy-feedback VP showed higher empathy in the SP interview than did the students in the backstory VP and the control VP groups [mean (SD) empathy scores coded on a 0-6 scale were 2.91 (0.16) vs. 2.20 (0.22) and 2.27 (0.21), respectively). The difference in scores was significant only for the empathy-feedback VP versus the backstory VP group (P = 0.027). The SPs rated the empathy-feedback and the backstory VP groups significantly higher than the control VP group on offering empathic statements (P < 0.0001), appearing warm and caring (P = 0.015), and forming rapport (P = 0.004). Feedback on empathy in a VP interaction increased students' empathy in encounters with SPs, as rated by trained assessors, whereas a simulation of patient shadowing did not. Both VP interventions increased students' empathy as rated by SPs, compared with the control VP group.

Genetic Diversity of Hepatitis A Virus in China: VP3-VP1-2A Genes and Evidence of Quasispecies Distribution in the Isolates

PubMed Central

Cao, Jingyuan; Zhou, Wenting; Yi, Yao; Jia, Zhiyuan; Bi, Shengli

2013-01-01

Hepatitis A virus (HAV) is the most common cause of infectious hepatitis throughout the world, spread largely by the fecal-oral route. To characterize the genetic diversity of the virus circulating in China where HAV in endemic, we selected the outbreak cases with identical sequences in VP1-2A junction region and compiled a panel of 42 isolates. The VP3-VP1-2A regions of the HAV capsid-coding genes were further sequenced and analyzed. The quasispecies distribution was evaluated by cloning the VP3 and VP1-2A genes in three clinical samples. Phylogenetic analysis demonstrated that the same genotyping results could be obtained whether using the complete VP3, VP1, or partial VP1-2A genes for analysis in this study, although some differences did exist. Most isolates clustered in sub-genotype IA, and fewer in sub-genotype IB. No amino acid mutations were found at the published neutralizing epitope sites, however, several unique amino acid substitutions in the VP3 or VP1 region were identified, with two amino acid variants closely located to the immunodominant site. Quasispecies analysis showed the mutation frequencies were in the range of 7.22x10-4 -2.33x10-3 substitutions per nucleotide for VP3, VP1, or VP1-2A. When compared with the consensus sequences, mutated nucleotide sites represented the minority of all the analyzed sequences sites. HAV replicated as a complex distribution of closely genetically related variants referred to as quasispecies, and were under negative selection. The results indicate that diverse HAV strains and quasispecies inside the viral populations are presented in China, with unique amino acid substitutions detected close to the immunodominant site, and that the possibility of antigenic escaping mutants cannot be ruled out and needs to be further analyzed. PMID:24069343

Determinants of the VP1/2A junction cleavage by the 3C protease in foot-and-mouth disease virus-infected cells.

PubMed

Kristensen, Thea; Normann, Preben; Gullberg, Maria; Fahnøe, Ulrik; Polacek, Charlotta; Rasmussen, Thomas Bruun; Belsham, Graham J

2017-03-01

The foot-and-mouth disease virus (FMDV) capsid precursor, P1-2A, is cleaved by FMDV 3C protease to yield VP0, VP3, VP1 and 2A. Cleavage of the VP1/2A junction is the slowest. Serotype O FMDVs with uncleaved VP1-2A (having a K210E substitution in VP1; at position P2 in cleavage site) have been described previously and acquired a second site substitution (VP1 E83K) during virus rescue. Furthermore, introduction of the VP1 E83K substitution alone generated a second site change at the VP1/2A junction (2A L2P, position P2' in cleavage site). These virus adaptations have now been analysed using next-generation sequencing to determine sub-consensus level changes in the virus; this revealed other variants within the E83K mutant virus population that changed residue VP1 K210. The construction of serotype A viruses with a blocked VP1/2A cleavage site (containing K210E) has now been achieved. A collection of alternative amino acid substitutions was made at this site, and the properties of the mutant viruses were determined. Only the presence of a positively charged residue at position P2 in the cleavage site permitted efficient cleavage of the VP1/2A junction, consistent with analyses of diverse FMDV genome sequences. Interestingly, in contrast to the serotype O virus results, no second site mutations occurred within the VP1 coding region of serotype A viruses with the blocked VP1/2A cleavage site. However, some of these viruses acquired changes in the 2C protein that is involved in enterovirus morphogenesis. These results have implications for the testing of potential antiviral agents targeting the FMDV 3C protease.

Comparative characteristics of the VP7 and VP4 antigenic epitopes of the rotaviruses circulating in Russia (Nizhny Novgorod) and the Rotarix and RotaTeq vaccines.

PubMed

Morozova, O V; Sashina, T A; Fomina, S G; Novikova, N A

2015-07-01

Two live, attenuated rotavirus A (RVA) vaccines, Rotarix and RotaTeq, have been successfully introduced into national immunization programs worldwide. The parent strains of both vaccines were obtained more than 30 years ago. Nonetheless, only very limited data are available on the molecular similarity of the vaccine strains and their genetic relationships to the wild-type strains circulating within the territory of Russian Federation. In this study, we have determined the nucleotide sequences of the genes encoding the viral proteins VP7 and VP4 (the globular domain VP8*) of vaccine strains and natural isolates of rotaviruses in Nizhny Novgorod, Russia. The VP7 and VP4 proteins contain antigenic sites that are the main targets of neutralizing antibodies. Phylogenetic analysis based on VP4 and VP7 showed that the majority of the natural RVA isolates from Nizhny Novgorod and the vaccine strains belong to different clusters. Four amino acids within the VP7 antigenic sites were common in both the wild-type and vaccine strains. The largest number of amino acid differences was found between the vaccine strain Rotarix and the Nizhny Novgorod G2 strains (19 residues out of 29). From 3 to 5 amino acid differences per strain were identified in the antigenic sites of VP4 (domain VP8*) between wild-type strains and the vaccine RotaTeq, and 6-8 substitutions were found when they were compared with the vaccine strain Rotarix. For the first time, immunodominant T-cell epitopes of VP7 were analyzed, and differences in the sequences between the vaccine and the wild-type strains were found. The accumulation of amino acid substitutions in the VP7 and VP4 antigenic sites may potentially reduce the immune protection of vaccinated children from wild-type strains of rotavirus.

Clinical clues for differential diagnosis between verruca plana and verruca plana-like seborrheic keratosis.

PubMed

Kim, Won-Jeong; Lee, Won-Ku; Song, Margaret; Kim, Hoon-Soo; Ko, Hyun-Chang; Kim, Byung-Soo; Kim, Moon-Bum

2015-04-01

Sometimes the clinical differentiation between verruca plana (VP) and VP-like seborrheic keratosis (SK) could be challenged. However, there have been no studies on this issue to date. The aim of this study was to elucidate clinical and dermoscopic differences between these two diseases, and also to suggest a diagnostic algorithm of VP and VP-like SK without skin biopsy. The patients who had lesions clinically considered as VP or VP-like SK were the target of our study. We took clinical and dermoscopic photos with informed consent and conducted a questionnaire. All patients had their diagnoses confirmed by biopsy. Thirty-three patients were enrolled in our study. Seventeen patients were finally diagnosed with VP (51.5%) and 16 patients with VP-like SK (48.5%). In clinical findings, VP-like SK showed significantly more scattered distribution than VP (P = 0.039), which exhibited more clustered or grouped distribution (P = 0.039). In dermoscopic findings, brain-like appearance was more commonly observed in VP-like SK (P = 0.003) whereas VP showed more red dots or globular vessels (P = 0.017) and even-colored light brown to yellow patch (P < 0.001). Sex, onset age, the size of each lesion, location, color and shape showed no significant differences between them (P > 0.05). Based on our results, we suggest a diagnostic algorithm using Koebner's phenomenon, dermoscopic findings, distribution of each lesion and biopsy for multiple VP-like lesions in adults, and we think it will be a very useful diagnostic tool in daily clinical dermatological practice. © 2015 Japanese Dermatological Association.

Expression and immunogenic analysis of recombinant polypeptides derived from capsid protein VP1 for developing subunit vaccine material against hepatitis A virus.

PubMed

Jang, Kyoung Ok; Park, Jong-Hwa; Lee, Hyun Ho; Chung, Dae Kyun; Kim, Wonyong; Chung, In Sik

2014-08-01

Three recombinant polypeptides, VP1-His, VP1-3N-His, and 3D2-His, were produced by Escherichia coli expression system. Recombinant VP1-His, VP1-3N-His, and 3D2-His were expressed as bands with molecular weights of 32, 38, and 30 kDa, respectively. These were purified by affinity chromatography using Ni-NTA Fast-flow resin and/or ion-exchange chromatography using DEAE-Sepharose Fast-flow resin. Intraperitoneal immunizations of recombinant polypeptides successfully elicited the productions of VP1-His, VP1-3N-His, and 3D2-His specific IgG antibodies (IgG subclass distribution of IgG1>IgG2a>IgG2b>IgG3) in sera and induced the secretions of cytokines IFN-γ and IL-6 in spleen cells. Sera from recombinant VP1-His-, VP1-3N-His-, and 3D2-His-immunized mice neutralized the propagation of HAV. The highest neutralizing activity was shown in sera from recombinant VP1-3N-His-immunized mice. These results suggest that recombinant VP1-3N-His can be a useful source for developing hepatitis A virus (HAV) subunit vaccine candidates. Copyright © 2014 Elsevier Inc. All rights reserved.

Immunogenicity and protective efficacy of rotavirus VP8* fused to cholera toxin B subunit in a mouse model.

PubMed

Xue, Miaoge; Yu, Linqi; Jia, Lianzhi; Li, Yijian; Zeng, Yuanjun; Li, Tingdong; Ge, Shengxiang; Xia, Ningshao

2016-11-01

In attempts to develop recombinant subunit vaccines against rotavirus disease, it was previously shown that the N-terminal truncated VP8* protein, VP8-1 (aa26-231), is a good vaccine candidate when used for immunization in combination with Freund's adjuvant. However, this protein stimulated only weak immune response when aluminum hydroxide was used as an adjuvant. In this study, the nontoxic B subunit of cholera toxin (CTB) was employed as intra-molecular adjuvant to improve the immunogenicity of VP8-1. Both, the N-terminal and C-terminal fusion proteins, were purified to homogeneity, at which stage they formed pentamers, and showed significantly higher immunogenicity and protective efficacy than a VP8-1/aluminum hydroxide mixture in a mouse model. Compared to VP8-1-CTB, CTB-VP8-1 showed higher binding activity to both, GM1 and the conformation sensitive neutralizing monoclonal antibodies specific to VP8. More importantly, CTB-VP8-1 elicited higher titers of neutralizing antibodies and conferred higher protective efficacy than VP8-1-CTB. Therefore, the protein CTB-VP8-1, with enhanced immunogenicity and immunoprotectivity, could be considered as a viable candidate for further development of an alternative, replication-incompetent, parenterally administered vaccine against rotavirus disease.

Immunogenicity of virus-like particles containing modified goose parvovirus VP2 protein.

PubMed

Chen, Zongyan; Li, Chuanfeng; Zhu, Yingqi; Wang, Binbin; Meng, Chunchun; Liu, Guangqing

2012-10-01

The major capsid protein VP2 of goose parvovirus (GPV) expressed using a baculovirus expression system (BES) assembles into virus-like particles (VLPs). To optimize VP2 gene expression in Sf9 cells, we converted wild-type VP2 (VP2) codons into codons that are more common in insect genes. This change greatly increased VP2 protein production in Sf9 cells. The protein generated from the codon-optimized VP2 (optVP2) was detected by immunoblotting and an indirect immunofluorescence assay (IFA). Transmission electron microscopy analysis revealed the formation of VLPs. These findings indicate that optVP2 yielded stable and high-quality VLPs. Immunogenicity assays revealed that the VLPs are highly immunogenic, elicit a high level of neutralizing antibodies and provide protection against lethal challenge. The antibody levels appeared to be directly related to the number of GP-Ag-positive hepatocytes. The variation trends for GP-Ag-positive hepatocytes were similar in the vaccine groups. In comparison with the control group, the optVP2 VLPs groups exhibited obviously better responses. These data indicate that the VLPs retained immunoreactivity and had strong immunogenicity in susceptible geese. Thus, GPV optVP2 appears to be a good candidate for the vaccination of goslings. Copyright © 2012 Elsevier B.V. All rights reserved.

Surface modification of silicon wafer by grafting zwitterionic polymers to improve its antifouling property

NASA Astrophysics Data System (ADS)

Sun, Yunlong; Chen, Changlin; Xu, Heng; Lei, Kun; Xu, Guanzhe; Zhao, Li; Lang, Meidong

2017-10-01

Silicon (111) wafer was modified by triethoxyvinylsilane containing double bond as an intermedium, and then P4VP (polymer 4-vinyl pyridine) brush was "grafted" onto the surface of silicon wafer containing reactive double bonds by adopting the "grafting from" way and Si-P4VP substrate (silicon wafer grafted by P4VP) was obtained. Finally, P4VP brush of Si-P4VP substrate was modified by 1,3-propanesulfonate fully to obtain P4VP-psl brush (zwitterionic polypyridinium salt) and the functional Si-P4VP-psl substrate (silicon wafer grafted by zwitterionic polypyridinium salt based on polymer 4-vinyl pyridine) was obtained successfully. The antifouling property of the silicon wafer, the Si-P4VP substrate and the Si-P4VP-psl substrate was investigated by using bovine serum albumin, mononuclear macrophages (RAW 264.7) and Escherichia coli (E. coli) ATTC25922 as model bacterium. The results showed that compared with the blank sample-silicon wafer, the Si-P4VP-psl substrate had excellent anti-adhesion ability against bovine serum albumin, cells and bacterium, due to zwitterionic P4VP-psl brush (polymer 4-vinyl pyridine salt) having special functionality like antifouling ability on biomaterial field.

Subunit Vaccine Preparation of Bovine Rotavirus and Its Efficacy in Mice.

PubMed

Suocheng, Wei; Tuanjie, Che; Changjun, Song; Fengling, Tian; Zhongren, Ma

2015-09-01

Rotaviruses (RV) are important viral diarrheal agents in calves. Vaccination is an optimum measure to prevent bovine rotaviruses (BRV) infection. However, little research on BRV VP7 vaccine has been done and currently there is no BRV vaccine. To prepare a subunit vaccine of BRV and investigate its efficacy. Total RNA was extracted from MA104 cells infected with bovine rotavirus (BRV) strain GSB01. BRV VP7 gene was amplified using real time fluorescence quantitative PCR (qPCR). The pEASY-T3-VP7 plasmid was digested using HindⅢ and BamHI restriction endonucleases, then recombined into the prokaryotic expression vector pET32a. The pET32a-VP7 and pET32a-VP7-LTB (heat-labile enterotoxin B subunit) were transformed into BL21 (DE3) competent cells of Escherichia coli, respectively, and induced with IPTG, then analyzed using SDS-PAGE. Sixty mice were randomly divided into three groups (n=20). Group A mice was used as His-tag control and mice in group B and C were inoculated with pET32a-VP7 and pET32a-VP7-LTB, respectively. VP7 IgG antibody titers and protection efficiency of pET32a-VP7-LTB were further determined in neonatal mice challenged with GSB01 BRV strain. SDS-PAGE analysis showed that the pET32a-VP7 was highly expressed in the BL21 (DE3) cells. PET32a-VP7 and pET32a-VP7-LTB protein could promote VP7 IgG antibody titer（8.33×103 vs. 17.26×103）in mice. Immunization protection ratios of pET32a-VP7 and pET32a-VP7-LTB proteins in the neonatal mice were 86.4% and 91.7%, respectively. The fusion protein of pET32a-VP7-LTB had excellent immunogenicity and protected mice from BRV infection. Our findings can be used for further developing of a high-efficiency subunit vaccine of BRV.

Utilization of fluorescence spectroscopy as a novel approach to evaluate the oxidative stability in beef retail displayed.

PubMed

Pouzo, L B; Zaritzky, N E; Pavan, E; Rossetti, L; Descalzo, A M

2016-09-01

Beef samples from grazing steers finished with different seed-supplemented diets were vacuum packaged for 3, 14 and 56days (VC) and subsequently exposed to aerobic conditions (AE) for 0 and 5days. Different fluorescent compounds of interest in the oxidation process were detected in meat, namely tryptophan residues, Schiff bases and porphyrins. Tryptophan intensity fluorescence increased with 14days of VC; while Schiff bases intensity increased (P<0.05) in beef samples stored under VC-56 and in all samples after AE-5days. Porphyrins increased (P<0.05) gradually with the extension of vacuum storage time, but were degraded in beef with long vacuum storage and 5days of AE. Higher levels of porphyrins in beef under VC were correlated (P<0.05) with lower redness and higher TBARS after AE-5. This study revealed the potential of fluorescence signals to detect oxidative changes in beef under different storage conditions using a fast and nondestructive method such as fluorescence spectroscopy. Copyright © 2016 Elsevier Ltd. All rights reserved.

Microbiological safety of retail vacuum-packed and modified-atmosphere-packed cooked meats at end of shelf life.

PubMed

Sagoo, S K; Little, C L; Allen, G; Williamson, K; Grant, K A

2007-04-01

A study of retail modified-atmosphere-packed and vacuum-packed cooked ready-to-eat meats was undertaken from September through mid-November 2003 to determine the microbiological quality at the end of shelf life and to establish any risk factors in the production, storage, and display of this product. Examination of 2,981 samples using Microbiological Guidelines criteria revealed that 66% were of satisfactory or acceptable microbiology quality, 33% were of unsatisfactory quality mainly due to high aerobic colony counts and Enterobacteriaceae concentrations, and 1% were of unacceptable quality due to the presence of Listeria monocytogenes at 100 CFU/g or higher (27 samples; range of 10(2) to 106 CFU/g) and Campylobacter jejuni (1 sample), indicating a risk to health. All samples at the end of the shelf life had satisfactory (<20 CFU/g) and/or acceptable (<102 CFU/g) levels of Staphylococcus aureus and Clostridium perfringens, four samples (<1%) had unsatisfactory levels of Escherichia coli (range of 102 to 106 CFU/g) and 5.5% of the samples contained L. monocytogenes at <20 CFU/g (4.8%) or between 20 and 100 CFU/g (0.7%). More samples of chicken (45%; 224 of 495 samples), beef (43%; 160 of 371 samples), and turkey (41%; 219 of 523 samples) were of unsatisfactory or unacceptable quality compared with ham (23%; 317 of 1,351 samples) or pork (32%; 67 of 206 samples). Twelve different L. monocytogenes typing characters (serotype-amplified fragment length polymorphism type-phage type) were evaluated for isolates recovered from samples of unacceptable quality, and the 1/2-IX-NT type was recovered from almost half (48%) of these samples. Salmonella was not detected in any samples examined. Risk factors identified for cooked meats that were microbiologically contaminated more frequently included vacuum packaging, packaging on retail premises, slicing, temperature not monitored in display units, and no hazard analysis system in place. Results from this study also suggest that the shelf life assigned to some modified-atmosphere-packed and vacuum-packed meats may not be appropriate.

Dynamic phosphorylation of Ebola virus VP30 in NP-induced inclusion bodies.

PubMed

Lier, Clemens; Becker, Stephan; Biedenkopf, Nadine

2017-12-01

Zaire Ebolavirus (EBOV) causes a severe feverish disease with high case fatality rates. Transcription of EBOV is dependent on the activity of the nucleocapsid protein VP30 which represents an essential viral transcription factor. Activity of VP30 is regulated via phosphorylation at six N-terminal serine residues. Recent data demonstrated that dynamic phosphorylation and dephosphorylation of serine residue 29 is essential for transcriptional support activity of VP30. To analyze the spatio/temporal dynamics of VP30 phosphorylation, we generated a peptide antibody recognizing specifically VP30 phosphorylated at serine 29. Using this antibody we could demonstrate that (i) the majority of VP30 molecules in EBOV-infected cells is dephosphorylated at the crucial position serine 29, (ii) both, VP30 phosphorylation and dephosphorylation take place in viral inclusion bodies that are induced by the nucleoprotein NP and (iii) NP influences the phosphorylation state of VP30. Copyright © 2017 Elsevier Inc. All rights reserved.

First study conducted in Northern India that identifies group C rotavirus as the etiological agent of severe diarrhea in children in Delhi.

PubMed

Tiku, Vasundhara Razdan; Jiang, Baoming; Kumar, Praveen; Aneja, Satender; Bagga, Arvind; Bhan, Maharaj Kishen; Ray, Pratima

2017-05-30

Group C Rotavirus (RVC) is an enteric pathogen responsible for acute gastroenteritis in children and adults globally. At present there are no surveillance studies on group C Rotaviruses in India and therefore their prevalence in India remains unknown. The present study aimed to evaluate group C rotavirus infection among <5 years old children hospitalized with acute gastroenteritis in New Delhi. A total of 350 fecal specimens were collected during September 2013 to November 2014 from <5 years old diarrheal patients admitted at KSCH hospital, Delhi. The samples found negative for group A rotavirus (N = 180) by Enzyme immunoassay were screened for group C rotavirus by RT-PCR with VP6, VP7 and VP4 gene specific primers. The PCR products were further sequenced (VP6, VP7, VP4) and analyzed to ascertain their origin and G and P genotypes. Six out of 180 (group A rotavirus negative) samples were found positive for group C rotavirus by VP6 gene specific RT-PCR, of which 3 were also found positive for VP7 and VP4 genes. Phylogenetic analysis of VP7 and VP4 genes of these showed them to be G4 and P[2] genotypes. Overall, the nucleotide sequence data (VP6, VP7 and VP4) revealed a close relationship with the human group C rotavirus with no evidence of animal ancestry. Interestingly, the nucleotide sequence analysis of various genes also indicated differences in their origin. While the identity matrix of VP4 gene (n = 3) showed high amino acid sequence identity (97.60 to 98.20%) with Korean strain, the VP6 gene (n = 6) showed maximum identity with Nigerian strain (96.40 to 97.60%) and VP7 gene (n = 3) with Bangladeshi and USA strains. This is true for all analyzed samples. Our study demonstrated the group C rotavirus as the cause of severe diarrhea in young children in Delhi and provides insights on the origin of group C rotavirus genes among the local strains indicating their source of transmission. Our study also highlights the need for a simple and reliable diagnostic test that can be utilized to determine the disease burden due to group C rotavirus in India.

Seismic Instrument Development

DTIC Science & Technology

1990-01-01

xnxx .~.0722MICROMETER RP .025/REV. - .000681 8 INCHES PER MIN. WITH 5.4 RPM MOTOR. RATE IS .0006818 IN/MIN. FIGURE 2.1.1-1. STRAIN/INERTIAL GEAR...is located on the circuit board in the outer high-vacuum com - partment. The schematic of the transducer and transducer amplifier is shown in figure...packaged for shipment by com - mercial air carrier using standard commercial practices. The total system may be air lifted by helicopter to remote

Military Curricula for Vocational & Technical Education. Basic Electricity and Electronics. CANTRAC A-100-0010. Module 21: Basic Transistor Theory; Module 21T: Multi-Element Vacuum Tubes. Study Booklet.

ERIC Educational Resources Information Center

Chief of Naval Education and Training Support, Pensacola, FL.

This set of individualized learning modules on transistor theory is one in a series of modules for a course in basic electricity and electronics. The course is one of a number of military-developed curriculum packages selected for adaptation to vocational instructional and curriculum development in a civilian setting. Two modules are included in…

Radappertized Meats for Aerospace Meals

DTIC Science & Technology

1981-06-01

and canned bread made from irradiated wheat flour were used as emergency back-up foods in Skylab III. In response to a NASA request for foods for the...tt a,- /_ j .3 LIST OF TABLES PUge 1. Test Results of Vacuum Packaged Irradiated 10 Food Samples 2. Chemical Composition of Irradiated Food Samples...55 g Sodium nitrite .. ........ 1 g *Pickling spice mixture . . .... .... . 340 g (dry) The pumped briskets were stored at 30 C for 48 hours in

4-D Visualization of Seismic and Geodetic Data of the Big Island of Hawai'i

NASA Astrophysics Data System (ADS)

Burstein, J. A.; Smith-Konter, B. R.; Aryal, A.

2017-12-01

For decades Hawai'i has served as a natural laboratory for studying complex interactions between magmatic and seismic processes. Investigating characteristics of these processes, as well as the crustal response to major Hawaiian earthquakes, requires a synthesis of seismic and geodetic data and models. Here, we present a 4-D visualization of the Big Island of Hawai'i that investigates geospatial and temporal relationships of seismicity, seismic velocity structure, and GPS crustal motions to known volcanic and seismically active features. Using the QPS Fledermaus visualization package, we compile 90 m resolution topographic data from NASA's Shuttle Radar Topography Mission (SRTM) and 50 m resolution bathymetric data from the Hawaiian Mapping Research Group (HMRG) with a high-precision earthquake catalog of more than 130,000 events from 1992-2009 [Matoza et al., 2013] and a 3-D seismic velocity model of Hawai'i [Lin et al., 2014] based on seismic data from the Hawaiian Volcano Observatory (HVO). Long-term crustal motion vectors are integrated into the visualization from HVO GPS time-series data. These interactive data sets reveal well-defined seismic structure near the summit areas of Mauna Loa and Kilauea volcanoes, where high Vp and high Vp/Vs anomalies at 5-12 km depth, as well as clusters of low magnitude (M < 3.5) seismicity, are observed. These areas of high Vp and high Vp/Vs are interpreted as mafic dike complexes and the surrounding seismic clusters are associated with shallow magma processes. GPS data are also used to help identify seismic clusters associated with the steady crustal detachment of the south flank of Kilauea's East Rift Zone. We also investigate the fault geometry of the 2006 M6.7 Kiholo Bay earthquake event by analyzing elastic dislocation deformation modeling results [Okada, 1985] and HVO GPS and seismic data of this event. We demonstrate the 3-D fault mechanisms of the Kiholo Bay main shock as a combination of strike-slip and dip-slip components (net slip 0.55 m) delineating a 30 km east-west striking, southward-dipping fault plane, occurring at 39 km depth. This visualization serves as a resource for advancing scientific analyses of Hawaiian seismic processes, as well as an interactive educational tool for demonstrating the geospatial and geophysical structure of the Big Island of Hawai'i.

CdS/C60 binary nanocomposite films prepared via phase transition of PS-b-P2VP block copolymer.

PubMed

Lee, Jung-Pil; Koh, Haeng-Deog; Shin, Won-Jeong; Kang, Nam-Goo; Park, Soojin; Lee, Jae-Suk

2014-03-01

We demonstrate the well-defined control of phase transition of a polystyrene-b-poly(2-vinylpyridine) (PS-b-P2VP) block copolymer from spherical micelles to lamellar structures, in which CdS and C60 nanoparticles (NPs) are selectively positioned at the P2VP domains. The CdS NPs are in situ synthesized using PS-b-P2VP block copolymer templates that are self-assembled in PS-selective solvents. The CdS-PS-b-P2VP micellar structures are transformed to lamellar phase by adjusting a solvent selectivity for both blocks. In addition, a binary system of CdS/C60 embedded in PS-b-P2VP lamellar structures (CdS/C60-PS-b-P2VP) is fabricated by embedding C60 molecules into P2VP domain though charge-transfer complexation between pyridine units of PS-b-P2VP and C60 molecules. The CdS/C60-PS-b-P2VP nanostructured films are characterized by transmission electron microscopy (TEM) and UV-Vis spectrometer. Copyright © 2013 Elsevier Inc. All rights reserved.

Molecular Tectonic Model of Virus Structural Transitions: the Putative Cell Entry States of Poliovirus

PubMed Central

Belnap, David M.; Filman, David J.; Trus, Benes L.; Cheng, Naiqian; Booy, Frank P.; Conway, James F.; Curry, Stephen; Hiremath, Chaitanya N.; Tsang, Simon K.; Steven, Alasdair C.; Hogle, James M.

2000-01-01

Upon interacting with its receptor, poliovirus undergoes conformational changes that are implicated in cell entry, including the externalization of the viral protein VP4 and the N terminus of VP1. We have determined the structures of native virions and of two putative cell entry intermediates, the 135S and 80S particles, at ∼22-Å resolution by cryo-electron microscopy. The 135S and 80S particles are both ∼4% larger than the virion. Pseudoatomic models were constructed by adjusting the beta-barrel domains of the three capsid proteins VP1, VP2, and VP3 from their known positions in the virion to fit the 135S and 80S reconstructions. Domain movements of up to 9 Å were detected, analogous to the shifting of tectonic plates. These movements create gaps between adjacent subunits. The gaps at the sites where VP1, VP2, and VP3 subunits meet are plausible candidates for the emergence of VP4 and the N terminus of VP1. The implications of these observations are discussed for models in which the externalized components form a transmembrane pore through which viral RNA enters the infected cell. PMID:10627545

Molecular tectonic model of virus structural transitions: the putative cell entry states of poliovirus.

PubMed

Belnap, D M; Filman, D J; Trus, B L; Cheng, N; Booy, F P; Conway, J F; Curry, S; Hiremath, C N; Tsang, S K; Steven, A C; Hogle, J M

2000-02-01

Upon interacting with its receptor, poliovirus undergoes conformational changes that are implicated in cell entry, including the externalization of the viral protein VP4 and the N terminus of VP1. We have determined the structures of native virions and of two putative cell entry intermediates, the 135S and 80S particles, at approximately 22-A resolution by cryo-electron microscopy. The 135S and 80S particles are both approximately 4% larger than the virion. Pseudoatomic models were constructed by adjusting the beta-barrel domains of the three capsid proteins VP1, VP2, and VP3 from their known positions in the virion to fit the 135S and 80S reconstructions. Domain movements of up to 9 A were detected, analogous to the shifting of tectonic plates. These movements create gaps between adjacent subunits. The gaps at the sites where VP1, VP2, and VP3 subunits meet are plausible candidates for the emergence of VP4 and the N terminus of VP1. The implications of these observations are discussed for models in which the externalized components form a transmembrane pore through which viral RNA enters the infected cell.

A novel oral delivery system consisting in "drug-in cyclodextrin-in nanostructured lipid carriers" for poorly water-soluble drug: vinpocetine.

PubMed

Lin, Congcong; Chen, Fen; Ye, Tiantian; Zhang, Lina; Zhang, Wenji; Liu, Dandan; Xiong, Wei; Yang, Xinggang; Pan, Weisan

2014-04-25

The purpose of this study was to develop a new delivery system based on drug cyclodextrin (CD) complexation and loading into nanostructured lipid carriers (NLC) to improve the oral bioavailability of vinpocetine (VP). Three different CDs and three different methods to obtain solid vinpocetine-cyclodextrin-tartaric acid complexes (VP-CD-TA) were contrasted. The co-evaporation vinpocetine-β-cyclodextrin-tartaric acid loaded NLC (VP-β-CD-TA COE-loaded NLC) was obtained by emulsification ultrasonic dispersion method. VP-β-CD-TA COE-loaded NLC was suitably characterized for particle size, polydispersity index, zeta potential, entrapment efficiency and the morphology. The crystallization of drug in VP-CD-TA and NLC was investigated by differential scanning calorimetry (DSC). The in vitro release study was carried out at pH 1.2, pH 6.8 and pH 7.4 medium. New Zealand rabbits were applied to investigate the pharmacokinetic behavior in vivo. The VP-β-CD-TA COE-loaded NLC presented a superior physicochemical property and selected to further study. In the in vitro release study, VP-β-CD-TA COE-loaded NLC exhibited a higher dissolution rate in the pH 6.8 and pH 7.4 medium than VP suspension and VP-NLC. The relative bioavailability of VP-β-CD-TA COE-loaded NLC was 592% compared with VP suspension and 92% higher than VP-NLC. In conclusion, the new formulation significantly improved bioavailability of VP for oral delivery, demonstrated a perspective way for oral delivery of poorly water-soluble drugs. Copyright © 2014 Elsevier B.V. All rights reserved.

Antigenic profile of African horse sickness virus serotype 4 VP5 and identification of a neutralizing epitope shared with bluetongue virus and epizootic hemorrhagic disease virus.

PubMed

Martínez-Torrecuadrada, J L; Langeveld, J P; Venteo, A; Sanz, A; Dalsgaard, K; Hamilton, W D; Meloen, R H; Casal, J I

1999-05-10

African horse sickness virus (AHSV) causes a fatal disease in horses. The virus capsid is composed of a double protein layer, the outermost of which is formed by two proteins: VP2 and VP5. VP2 is known to determine the serotype of the virus and to contain the neutralizing epitopes. The biological function of VP5, the other component of the capsid, is unknown. In this report, AHSV VP5, expressed in insect cells alone or together with VP2, was able to induce AHSV-specific neutralizing antibodies. Moreover, two VP5-specific monoclonal antibodies (MAbs) that were able to neutralize the virus in a plaque reduction assay were generated. To dissect the antigenic structure of AHSV VP5, the protein was cloned in Escherichia coli using the pET3 system. The immunoreactivity of both MAbs, and horse and rabbit polyclonal antisera, with 17 overlapping fragments from VP5 was analyzed. The most immunodominant region was found in the N-terminal 330 residues of VP5, defining two antigenic regions, I (residues 151-200) and II (residues 83-120). The epitopes were further defined by PEPSCAN analysis with 12mer peptides, which determined eight antigenic sites in the N-terminal half of the molecule. Neutralizing epitopes were defined at positions 85-92 (PDPLSPGE) for MAb 10AE12 and at 179-185 (EEDLRTR) for MAb 10AC6. Epitope 10AE12 is highly conserved between the different orbiviruses. MAb 10AE12 was able to recognize bluetongue virus VP5 and epizootic hemorrhagic disease virus VP5 by several techniques. These data will be especially useful for vaccine development and diagnostic purposes. Copyright 1999 Academic Press.

Marburg Virus VP24 Protein Relieves Suppression of the NF-κB Pathway Through Interaction With Kelch-like ECH-Associated Protein 1.

PubMed

Edwards, Megan R; Basler, Christopher F

2015-10-01

Marburg virus (MARV) is an emerging zoonotic pathogen that causes hemorrhagic fever. MARV VP24 (mVP24) protein interacts with the host cell protein Kelch-like-ECH-associated protein 1 (Keap1). Keap1 interacts with and promotes the degradation of IκB kinase β (IKKβ), a component of the IκB kinase (IKK) complex that regulates nuclear factor-κB (NF-κB) activity. We studied whether mVP24 could relieve Keap1 repression of the NF-κB pathway. Coimmunoprecipitation assays were used to examine the interaction between Keap1 and IKKβ in the presence of wild-type mVP24 and mutants of mVP24 defective for binding to Keap1. Western blotting was used to determine levels of IKKβ expression in the presence of Keap1 and mVP24. NF-κB promoter-luciferase assays were used to determine the effect of mVP24 on Keap1-induced repression of activity. Expression of wild-type mVP24 disrupted the interaction of IKKβ and Keap1, whereas weakly interacting and noninteracting mVP24 mutants did not disrupt the interaction between Keap1 and IKKβ. The interaction of mVP24 with Keap1 enhanced IKKβ levels in the presence of Keap1. The interaction of mVP24 with Keap1 also relieved Keap1 repression of NF-κB reporter activity. mVP24 can relieve Keap1 repression of the NF-κB pathway through its interaction with Keap1. © The Author 2015. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

The Acheta domesticus Densovirus, Isolated from the European House Cricket, Has Evolved an Expression Strategy Unique among Parvoviruses▿†

PubMed Central

Liu, Kaiyu; Li, Yi; Jousset, Françoise-Xavière; Zadori, Zoltan; Szelei, Jozsef; Yu, Qian; Pham, Hanh Thi; Lépine, François; Bergoin, Max; Tijssen, Peter

2011-01-01

The Acheta domesticus densovirus (AdDNV), isolated from crickets, has been endemic in Europe for at least 35 years. Severe epizootics have also been observed in American commercial rearings since 2009 and 2010. The AdDNV genome was cloned and sequenced for this study. The transcription map showed that splicing occurred in both the nonstructural (NS) and capsid protein (VP) multicistronic RNAs. The splicing pattern of NS mRNA predicted 3 nonstructural proteins (NS1 [576 codons], NS2 [286 codons], and NS3 [213 codons]). The VP gene cassette contained two VP open reading frames (ORFs), of 597 (ORF-A) and 268 (ORF-B) codons. The VP2 sequence was shown by N-terminal Edman degradation and mass spectrometry to correspond with ORF-A. Mass spectrometry, sequencing, and Western blotting of baculovirus-expressed VPs versus native structural proteins demonstrated that the VP1 structural protein was generated by joining ORF-A and -B via splicing (splice II), eliminating the N terminus of VP2. This splice resulted in a nested set of VP1 (816 codons), VP3 (467 codons), and VP4 (429 codons) structural proteins. In contrast, the two splices within ORF-B (Ia and Ib) removed the donor site of intron II and resulted in VP2, VP3, and VP4 expression. ORF-B may also code for several nonstructural proteins, of 268, 233, and 158 codons. The small ORF-B contains the coding sequence for a phospholipase A2 motif found in VP1, which was shown previously to be critical for cellular uptake of the virus. These splicing features are unique among parvoviruses and define a new genus of ambisense densoviruses. PMID:21775445

The Ebola virus VP35 protein binds viral immunostimulatory and host RNAs identified through deep sequencing.

PubMed

Dilley, Kari A; Voorhies, Alexander A; Luthra, Priya; Puri, Vinita; Stockwell, Timothy B; Lorenzi, Hernan; Basler, Christopher F; Shabman, Reed S

2017-01-01

Ebola virus and Marburg virus are members of the Filovirdae family and causative agents of hemorrhagic fever with high fatality rates in humans. Filovirus virulence is partially attributed to the VP35 protein, a well-characterized inhibitor of the RIG-I-like receptor pathway that triggers the antiviral interferon (IFN) response. Prior work demonstrates the ability of VP35 to block potent RIG-I activators, such as Sendai virus (SeV), and this IFN-antagonist activity is directly correlated with its ability to bind RNA. Several structural studies demonstrate that VP35 binds short synthetic dsRNAs; yet, there are no data that identify viral immunostimulatory RNAs (isRNA) or host RNAs bound to VP35 in cells. Utilizing a SeV infection model, we demonstrate that both viral isRNA and host RNAs are bound to Ebola and Marburg VP35s in cells. By deep sequencing the purified VP35-bound RNA, we identified the SeV copy-back defective interfering (DI) RNA, previously identified as a robust RIG-I activator, as the isRNA bound by multiple filovirus VP35 proteins, including the VP35 protein from the West African outbreak strain (Makona EBOV). Moreover, RNAs isolated from a VP35 RNA-binding mutant were not immunostimulatory and did not include the SeV DI RNA. Strikingly, an analysis of host RNAs bound by wild-type, but not mutant, VP35 revealed that select host RNAs are preferentially bound by VP35 in cell culture. Taken together, these data support a model in which VP35 sequesters isRNA in virus-infected cells to avert RIG-I like receptor (RLR) activation.

Identification of a T-helper cell epitope on the rotavirus VP6 protein.

PubMed Central

Baños, D M; Lopez, S; Arias, C F; Esquivel, F R

1997-01-01

In this work, we have studied the T-helper (Th)-cell response against rotavirus, in a mouse model. Adult BALB/c mice were inoculated parenterally with porcine rotavirus YM, and the Th-cell response from spleen cells against the virus and two overlapping fragments of the major capsid protein VP6 (VP6(1-192) and VP6(171-397)) were evaluated in vitro. The Th cells recognized the YM virus and the two protein fragments, suggesting that there are at least two Th-cell epitopes on the VP6 molecule. To study the specificity of Th cells against VP6 at the clonal level, we established two Th-cell hybridomas cross-reactive for the VP6 protein of rotavirus strains YM and SA11. Both hybridomas recognized the VP6(171-397) polypeptide, and a synthetic peptide comprising the amino acids 289 to 302 (RLSFQLVRPPNMTP) of YM VP6 in the context of the major histocompatibility complex class II IEd molecule. The Th-cell hybridomas recognized rotavirus VP6 in a highly cross-reactive fashion, since they could be stimulated by eight different strains of rotavirus, including the murine rotavirus EDIM, that represent five G serotypes and at least two subgroups. The amino acid sequence of the VP6 epitope is highly conserved in most group A rotavirus strains sequenced so far. On the other hand, it was found that Th cells specific for the VP6 epitope may constitute an important proportion of the total polyclonal Th-cell response against rotavirus YM in spleen cells. These results demonstrate that VP6 can be a target for highly cross-reactive Th cells. PMID:8985366

Genetic diversity of G1P[8] rotavirus VP7 and VP8* antigens in Finland over a 20-year period: No evidence for selection pressure by universal mass vaccination with RotaTeq® vaccine.

PubMed

Hemming, Maria; Vesikari, Timo

2013-10-01

Two live-attenuated oral vaccines (Rotarix™ and Rotateq®) against rotavirus gastroenteritis were licensed in 2006 and have been introduced into National Immunization Programs (NIPs) of several countries. Large scale use of rotavirus vaccines might cause antigenic pressure on circulating rotavirus types or lead to selection of new rotaviruses thus decreasing vaccine efficacy. We examined the nucleotide and amino acid sequences of the surface proteins VP7 and VP4 (cleaved to VP8(*) and VP5(*)) of a total of 108 G1P[8] rotavirus strains collected over a 20-year period from 1992, including the years 2006-2009 when rotavirus vaccine (mainly Rotarix™) was available, and the years 2009-2012 after implementation of RotaTeq® vaccine into the NIP of Finland. In G1 VP7 no changes at amino acid level were observed. In VP8(*) periodical fluctuation of the sublineage over the study period was found with multiple changes both at nucleotide and amino acid levels. Most amino acid changes were in the dominant antigenic epitopes of VP8(*). A change in VP8(*) sublineage occurred between 2008 and 2009, with a temporal correlation to the use of Rotarix™ up to 30% coverage in the period. In contrast, no antigenic changes in the VP8(*) protein appeared to be correlated to the exclusive use of RotaTeq® vaccine after 2009. Nevertheless, long-term surveillance of antigenic changes in VP4 and also VP7 proteins in wild-type rotavirus strains is warranted in countries with large scale use of the currently licensed live oral rotavirus vaccines. Copyright © 2013 Elsevier B.V. All rights reserved.

Alphavirus Replicon DNA Vectors Expressing Ebola GP and VP40 Antigens Induce Humoral and Cellular Immune Responses in Mice

PubMed Central

Ren, Shoufeng; Wei, Qimei; Cai, Liya; Yang, Xuejing; Xing, Cuicui; Tan, Feng; Leavenworth, Jianmei W.; Liang, Shaohui; Liu, Wenquan

2018-01-01

Ebola virus (EBOV) causes severe hemorrhagic fevers in humans, and no approved therapeutics or vaccine is currently available. Glycoprotein (GP) is the major protective antigen of EBOV, and can generate virus-like particles (VLPs) by co-expression with matrix protein (VP40). In this study, we constructed a recombinant Alphavirus Semliki Forest virus (SFV) replicon vector DREP to express EBOV GP and matrix viral protein (VP40). EBOV VLPs were successfully generated and achieved budding from 293 cells after co-transfection with DREP-based GP and VP40 vectors (DREP-GP+DREP-VP40). Vaccination of BALB/c mice with DREP-GP, DREP-VP40, or DREP-GP+DREP-VP40 vectors, followed by immediate electroporation resulted in a mixed IgG subclass production, which recognized EBOV GP and/or VP40 proteins. This vaccination regimen also led to the generation of both Th1 and Th2 cellular immune responses in mice. Notably, vaccination with DREP-GP and DREP-VP40, which produces both GP and VP40 antigens, induced a significantly higher level of anti-GP IgG2a antibody and increased IFN-γ secreting CD8+ T-cell responses relative to vaccination with DREP-GP or DREP-VP40 vector alone. Our study indicates that co-expression of GP and VP40 antigens based on the SFV replicon vector generates EBOV VLPs in vitro, and vaccination with recombinant DREP vectors containing GP and VP40 antigens induces Ebola antigen-specific humoral and cellular immune responses in mice. This novel approach provides a simple and efficient vaccine platform for Ebola disease prevention. PMID:29375526

A cationic, C-terminal patch and structural rearrangements in Ebola virus matrix VP40 protein control its interactions with phosphatidylserine.

PubMed

Del Vecchio, Kathryn; Frick, Cary T; Gc, Jeevan B; Oda, Shun-Ichiro; Gerstman, Bernard S; Saphire, Erica Ollmann; Chapagain, Prem P; Stahelin, Robert V

2018-03-02

Ebola virus (EBOV) is a filamentous lipid-enveloped virus that causes hemorrhagic fever with a high fatality rate. Viral protein 40 (VP40) is the major EBOV matrix protein and regulates viral budding from the plasma membrane. VP40 is a transformer/morpheein that can structurally rearrange its native homodimer into either a hexameric filament that facilitates viral budding or an RNA-binding octameric ring that regulates viral transcription. VP40 associates with plasma-membrane lipids such as phosphatidylserine (PS), and this association is critical to budding from the host cell. However, it is poorly understood how different VP40 structures interact with PS, what essential residues are involved in this association, and whether VP40 has true selectivity for PS among different glycerophospholipid headgroups. In this study, we used lipid-binding assays, MD simulations, and cellular imaging to investigate the molecular basis of VP40-PS interactions and to determine whether different VP40 structures ( i.e. monomer, dimer, and octamer) can interact with PS-containing membranes. Results from quantitative analysis indicated that VP40 associates with PS vesicles via a cationic patch in the C-terminal domain (Lys 224, 225 and Lys 274, 275 ). Substitutions of these residues with alanine reduced PS-vesicle binding by >40-fold and abrogated VP40 localization to the plasma membrane. Dimeric VP40 had 2-fold greater affinity for PS-containing membranes than the monomer, whereas binding of the VP40 octameric ring was reduced by nearly 10-fold. Taken together, these results suggest the different VP40 structures known to form in the viral life cycle harbor different affinities for PS-containing membranes. © 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

Optimizing CT technique to reduce radiation dose: effect of changes in kVp, iterative reconstruction, and noise index on dose and noise in a human cadaver.

PubMed

Chang, Kevin J; Collins, Scott; Li, Baojun; Mayo-Smith, William W

2017-06-01

For assessment of the effect of varying the peak kilovoltage (kVp), the adaptive statistical iterative reconstruction technique (ASiR), and automatic dose modulation on radiation dose and image noise in a human cadaver, a cadaver torso underwent CT scanning at 80, 100, 120 and 140 kVp, each at ASiR settings of 0, 30 and 50 %, and noise indices (NIs) of 5.5, 11 and 22. The volume CT dose index (CTDI vol ), image noise, and attenuation values of liver and fat were analyzed for 20 data sets. Size-specific dose estimates (SSDEs) and liver-to-fat contrast-to-noise ratios (CNRs) were calculated. Values for different combinations of kVp, ASiR, and NI were compared. The CTDI vol varied by a power of 2 with kVp values between 80 and 140 without ASiR. Increasing ASiR levels allowed a larger decrease in CTDI vol and SSDE at higher kVp than at lower kVp while image noise was held constant. In addition, CTDI vol and SSDE decreased with increasing NI at each kVp, but the decrease was greater at higher kVp than at lower kVp. Image noise increased with decreasing kVp despite a fixed NI; however, this noise could be offset with the use of ASiR. The CT number of the liver remained unchanged whereas that of fat decreased as the kVp decreased. Image noise and dose vary in a complicated manner when the kVp, ASiR, and NI are varied in a human cadaver. Optimization of CT protocols will require balancing of the effects of each of these parameters to maximize image quality while minimizing dose.

The Ebola virus VP35 protein binds viral immunostimulatory and host RNAs identified through deep sequencing

PubMed Central

Voorhies, Alexander A.; Luthra, Priya; Puri, Vinita; Stockwell, Timothy B.; Lorenzi, Hernan; Basler, Christopher F.; Shabman, Reed S.

2017-01-01

Ebola virus and Marburg virus are members of the Filovirdae family and causative agents of hemorrhagic fever with high fatality rates in humans. Filovirus virulence is partially attributed to the VP35 protein, a well-characterized inhibitor of the RIG-I-like receptor pathway that triggers the antiviral interferon (IFN) response. Prior work demonstrates the ability of VP35 to block potent RIG-I activators, such as Sendai virus (SeV), and this IFN-antagonist activity is directly correlated with its ability to bind RNA. Several structural studies demonstrate that VP35 binds short synthetic dsRNAs; yet, there are no data that identify viral immunostimulatory RNAs (isRNA) or host RNAs bound to VP35 in cells. Utilizing a SeV infection model, we demonstrate that both viral isRNA and host RNAs are bound to Ebola and Marburg VP35s in cells. By deep sequencing the purified VP35-bound RNA, we identified the SeV copy-back defective interfering (DI) RNA, previously identified as a robust RIG-I activator, as the isRNA bound by multiple filovirus VP35 proteins, including the VP35 protein from the West African outbreak strain (Makona EBOV). Moreover, RNAs isolated from a VP35 RNA-binding mutant were not immunostimulatory and did not include the SeV DI RNA. Strikingly, an analysis of host RNAs bound by wild-type, but not mutant, VP35 revealed that select host RNAs are preferentially bound by VP35 in cell culture. Taken together, these data support a model in which VP35 sequesters isRNA in virus-infected cells to avert RIG-I like receptor (RLR) activation. PMID:28636653

Different Temporal Effects of Ebola Virus VP35 and VP24 Proteins on Global Gene Expression in Human Dendritic Cells.

PubMed

Ilinykh, Philipp A; Lubaki, Ndongala M; Widen, Steven G; Renn, Lynnsey A; Theisen, Terence C; Rabin, Ronald L; Wood, Thomas G; Bukreyev, Alexander

2015-08-01

Ebola virus (EBOV) causes a severe hemorrhagic fever with a deficient immune response, lymphopenia, and lymphocyte apoptosis. Dendritic cells (DC), which trigger the adaptive response, do not mature despite EBOV infection. We recently demonstrated that DC maturation is unblocked by disabling the innate response antagonizing domains (IRADs) in EBOV VP35 and VP24 by the mutations R312A and K142A, respectively. Here we analyzed the effects of VP35 and VP24 with the IRADs disabled on global gene expression in human DC. Human monocyte-derived DC were infected by wild-type (wt) EBOV or EBOVs carrying the mutation in VP35 (EBOV/VP35m), VP24 (EBOV/VP24m), or both (EBOV/VP35m/VP24m). Global gene expression at 8 and 24 h was analyzed by deep sequencing, and the expression of interferon (IFN) subtypes up to 5 days postinfection was analyzed by quantitative reverse transcription-PCR (qRT-PCR). wt EBOV induced a weak global gene expression response, including markers of DC maturation, cytokines, chemokines, chemokine receptors, and multiple IFNs. The VP35 mutation unblocked the expression, resulting in a dramatic increase in expression of these transcripts at 8 and 24 h. Surprisingly, DC infected with EBOV/VP24m expressed lower levels of many of these transcripts at 8 h after infection, compared to wt EBOV. In contrast, at 24 h, expression of the transcripts increased in DC infected with any of the three mutants, compared to wt EBOV. Moreover, sets of genes affected by the two mutations only partially overlapped. Pathway analysis demonstrated that the VP35 mutation unblocked pathways involved in antigen processing and presentation and IFN signaling. These data suggest that EBOV IRADs have profound effects on the host adaptive immune response through massive transcriptional downregulation of DC. This study shows that infection of DC with EBOV, but not its mutant forms with the VP35 IRAD and/or VP24 IRAD disabled, causes a global block in expression of host genes. The temporal effects of mutations disrupting the two IRADs differ, and the lists of affected genes only partially overlap such that VP35 and VP24 IRADs each have profound effects on antigen presentation by exposed DC. The global modulation of DC gene expression and the resulting lack of their maturation represent a major mechanism by which EBOV disables the T cell response and suggests that these suppressive pathways are a therapeutic target that may unleash the T cell responses during EBOV infection. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Cryo-electron Microscopy Reconstruction and Stability Studies of the Wild Type and the R432A Variant of Adeno-associated Virus Type 2 Reveal that Capsid Structural Stability Is a Major Factor in Genome Packaging

PubMed Central

Drouin, Lauren M.; Lins, Bridget; Janssen, Maria; Bennett, Antonette; Chipman, Paul; McKenna, Robert; Chen, Weijun; Muzyczka, Nicholas; Cardone, Giovanni

2016-01-01

ABSTRACT The adeno-associated viruses (AAV) are promising therapeutic gene delivery vectors and better understanding of their capsid assembly and genome packaging mechanism is needed for improved vector production. Empty AAV capsids assemble in the nucleus prior to genome packaging by virally encoded Rep proteins. To elucidate the capsid determinants of this process, structural differences between wild-type (wt) AAV2 and a packaging deficient variant, AAV2-R432A, were examined using cryo-electron microscopy and three-dimensional image reconstruction both at an ∼5.0-Å resolution (medium) and also at 3.8- and 3.7-Å resolutions (high), respectively. The high resolution structures showed that removal of the arginine side chain in AAV2-R432A eliminated hydrogen bonding interactions, resulting in altered intramolecular and intermolecular interactions propagated from under the 3-fold axis toward the 5-fold channel. Consistent with these observations, differential scanning calorimetry showed an ∼10°C decrease in thermal stability for AAV2-R432A compared to wt-AAV2. In addition, the medium resolution structures revealed differences in the juxtaposition of the less ordered, N-terminal region of their capsid proteins, VP1/2/3. A structural rearrangement in AAV2-R432A repositioned the βA strand region under the icosahedral 2-fold axis rather than antiparallel to the βB strand, eliminating many intramolecular interactions. Thus, a single amino acid substitution can significantly alter the AAV capsid integrity to the extent of reducing its stability and possibly rendering it unable to tolerate the stress of genome packaging. Furthermore, the data show that the 2-, 3-, and 5-fold regions of the capsid contributed to producing the packaging defect and highlight a tight connection between the entire capsid in maintaining packaging efficiency. IMPORTANCE The mechanism of AAV genome packaging is still poorly understood, particularly with respect to the capsid determinants of the required capsid-Rep interaction. Understanding this mechanism may aid in the improvement of AAV packaging efficiency, which is currently ∼1:10 (10%) genome packaged to empty capsid in vector preparations. This report identifies regions of the AAV capsid that play roles in genome packaging and that may be important for Rep recognition. It also demonstrates the need to maintain capsid stability for the success of this process. This information is important for efforts to improve AAV genome packaging and will also inform the engineering of AAV capsid variants for improved tropism, specific tissue targeting, and host antibody escape by defining amino acids that cannot be altered without detriment to infectious vector production. PMID:27440903

Impact of Nisin-Activated Packaging on Microbiota of Beef Burgers during Storage

PubMed Central

Ferrocino, Ilario; Greppi, Anna; La Storia, Antonietta; Rantsiou, Kalliopi; Ercolini, Danilo

2015-01-01

Beef burgers were stored at 4°C in a vacuum in nisin-activated antimicrobial packaging. Microbial ecology analyses were performed on samples collected between days 0 and 21 of storage to discover the population diversity. Two batches were analyzed using RNA-based denaturing gradient gel electrophoresis (DGGE) and pyrosequencing. The active packaging retarded the growth of the total viable bacteria and lactic acid bacteria. Culture-independent analysis by pyrosequencing of RNA extracted directly from meat showed that Photobacterium phosphoreum, Lactococcus piscium, Lactobacillus sakei, and Leuconostoc carnosum were the major operational taxonomic units (OTUs) shared between control and treated samples. Beta diversity analysis of the 16S rRNA sequence data and RNA-DGGE showed a clear separation between two batches based on the microbiota. Control samples from batch B showed a significant high abundance of some taxa sensitive to nisin, such as Kocuria rhizophila, Staphylococcus xylosus, Leuconostoc carnosum, and Carnobacterium divergens, compared to control samples from batch A. However, only from batch B was it possible to find a significant difference between controls and treated samples during storage due to the active packaging. Predicted metagenomes confirmed differences between the two batches and indicated that the use of nisin-based antimicrobial packaging can determine a reduction in the abundance of specific metabolic pathways related to spoilage. The present study aimed to assess the viable bacterial communities in beef burgers stored in nisin-based antimicrobial packaging, and it highlights the efficacy of this strategy to prolong beef burger shelf life. PMID:26546424

A pragmatic approach to determine the optimal kVp in cone beam CT: balancing contrast-to-noise ratio and radiation dose

PubMed Central

Silkosessak, O; Jacobs, R; Bogaerts, R; Bosmans, H; Panmekiate, S

2014-01-01

Objectives: To determine the optimal kVp setting for a particular cone beam CT (CBCT) device by maximizing technical image quality at a fixed radiation dose. Methods: The 3D Accuitomo 170 (J. Morita Mfg. Corp., Kyoto, Japan) CBCT was used. The radiation dose as a function of kVp was measured in a cylindrical polymethyl methacrylate (PMMA) phantom using a small-volume ion chamber. Contrast-to-noise ratio (CNR) was measured using a PMMA phantom containing four materials (air, aluminium, polytetrafluoroethylene and low-density polyethylene), which was scanned using 180 combinations of kVp/mA, ranging from 60/1 to 90/8. The CNR was measured for each material using PMMA as background material. The pure effect of kVp and mAs on the CNR values was analysed. Using a polynomial fit for CNR as a function of mA for each kVp value, the optimal kVp was determined at five dose levels. Results: Absorbed doses ranged between 0.034 mGy mAs−1 (14 × 10 cm, 60 kVp) and 0.108 mGy mAs−1 (14 × 10 cm, 90 kVp). The relation between kVp and dose was quasilinear (R2 > 0.99). The effect of mA and kVp on CNR could be modelled using a second-degree polynomial. At a fixed dose, there was a tendency for higher CNR values at increasing kVp values, especially at low dose levels. A dose reduction through mA was more efficient than an equivalent reduction through kVp in terms of image quality deterioration. Conclusions: For the investigated CBCT model, the most optimal contrast at a fixed dose was found at the highest available kVp setting. There is great potential for dose reduction through mA with a minimal loss in image quality. PMID:24708447

Study of the Nankai seismogenic fault using dynamic wave propagation modelling of digital rock from the Nobeoka Fault

NASA Astrophysics Data System (ADS)

Eng, Chandoeun; Ikeda, Tatsunori; Tsuji, Takeshi

2018-10-01

To understand the characteristics of the Nankai seismogenic fault in the plate convergent margin, we calculated the P- and S-wave velocities (VP and VS) of digital rock models constructed from core samples of an ancient plate boundary fault at Nobeoka, Kyushu Island, Japan. We first constructed 3D digital rock models from microcomputed tomography images and identified their heterogeneous textures such as cracks and veins. We replaced the cracks and veins with air, water, quartz, calcite and other materials with different bulk and shear moduli. Using the Rotated Staggered Grid Finite-Difference Method, we performed dynamic wave propagation simulations and quantified the effective VP, VS and the ratio of VP to VS (VP/VS) of the 3D digital rock models with different crack-filling minerals. Our results demonstrate that the water-saturated cracks considerably decreased the seismic velocity and increased VP/VS. The VP/VS of the quartz-filled rock model was lower than that in the water-saturated case and in the calcite-filled rock model. By comparing the elastic properties derived from the digital rock models with the seismic velocities (e.g. VP and VP/VS) around the seismogenic fault estimated from field seismic data, we characterised the evolution process of the deep seismogenic fault. The high VP/VS and low VP observed at the transition from aseismic to coseismic regimes in the Nankai Trough can be explained by open cracks (or fractures), while the low VP/VS and high VP observed at the deeper coseismic fault zone suggests quartz-filled cracks. The quartz-rich fault zone characterised as low VP/VS and high VP in this study could partially relate to the coseismic behaviour as suggested by previous studies, because quartz exhibits slip-weakening behaviour (i.e. unstable coseismic slip).

[Effects of canine IL-2 and IL-7 genes on enhancing immunogenicity of canine parvovirus VP2 gene vaccine in mice].

PubMed

Chen, Huihui; Zhong, Fei; Li, Xiujin; Wang, Lu; Sun, Yan; Neng, Changai; Zhang, Kao; Li, Wenyan; Wen, Jiexia

2012-11-04

To investigate the effects of canine interleukin-2 (cIL-2) and cIL-7 genes on enhancing the immunogenicity of canine parvovirus (CPV) VP2 DNA vaccine. The bicistronic vectors of cIL-2 and cIL-7 genes were constructed using the eukaryotic expression vector containing internal ribosome entry site (IRES). The cIL-2/ cIL-7 dicistronic vector plus previously constructed vectors, including CPV VP2 DNA vaccine vector, cIL-2 vector and cIL-7 vector, were used to co-immunize mice with different combinations, consisting of VP2 alone, VP2 + cIL-2, VP2 + cIL-7 and VP2 + cIL-2/cIL-7. The VP2-specific antibody levels in immunized mice were measured by ELISA at different time post-immunization. The proliferation indices and interferon-gamma expression were measured by lymphocyte proliferation assay and ELISA, respectively. The cIL-2/cIL-7 bicistronic vector was correct and could mediate cIL-2 and cIL-7 gene expression in eukaryotic cells. Immunization results revealed that the antibody titers and the neutralizing antibody levels of the mice co-immunized with VP2 + cIL-7/cIL-2 vectors were significantly higher than that with either VP2 + cIL-2 vectors or VP2 + cIL-7 vectors (P < 0.05). The lymphocyte proliferation indices of VP2 + cIL-7/cIL-2 vector-immunized mice were also higher than that of other two groups although not statistically significant. However, the IFN-gamma expression levels of VP2 + cIL-7/cIL-2 vector-immunized mice were significantly higher than other immunized mice (P < 0.05). The cIL-2 and cIL-7 genes showed the significant synergic effects on enhancing the immunogenecity of CPV VP2 DNA vaccine.

Purification of full-length VP22 from cells infected with HSV-1: A two-pronged approach for the solubilization and purification of viral proteins for use in biochemical studies

PubMed Central

Dewberry, Ebony J.; Dunkerley, Eric; Duffy, Carol

2012-01-01

Summary VP22, encoded by the UL49 gene, is one of the most abundant proteins of the herpes simplex virus type 1 (HSV-1) tegument and has been shown to be important for virus replication and spread. However, the exact role(s) played by VP22 in the HSV-1 replication cycle have yet to be delineated. The lack of a procedure to purify full-length VP22 has limited molecular studies on VP22 function. A procedure was developed for the purification of soluble, full-length VP22 from cells infected with HSV-1. A recombinant virus encoding His-tagged VP22 was generated and found to express VP22 at levels comparable to the wild type virus upon infection of Vero cells. By experimenting with a wide variety of cell lysis buffer conditions, several buffers that promote the solubility of full-length VP22 were identified. Buffers that gave the highest levels of solubility were then used in immobilized metal ion affinity chromatography experiments to identify conditions that provided the greatest level of VP22 binding and recovery from cobalt and nickel affinity resins. Using this strategy soluble, full-length VP22 was purified from cells infected with HSV-1. PMID:22569534

Interaction of the Mouse Polyomavirus Capsid Proteins with Importins Is Required for Efficient Import of Viral DNA into the Cell Nucleus.

PubMed

Soldatova, Irina; Prilepskaja, Terezie; Abrahamyan, Levon; Forstová, Jitka; Huérfano, Sandra

2018-03-31

The mechanism used by mouse polyomavirus (MPyV) overcomes the crowded cytosol to reach the nucleus has not been fully elucidated. Here, we investigated the involvement of importin α/β1 mediated transport in the delivery of MPyV genomes into the nucleus. Interactions of the virus with importin β1 were studied by co-immunoprecipitation and proximity ligation assay. For infectivity and nucleus delivery assays, the virus and its capsid proteins mutated in the nuclear localization signals (NLSs) were prepared and produced. We found that at early times post infection, virions bound importin β1 in a time dependent manner with a peak of interactions at 6 h post infection. Mutation analysis revealed that only when the NLSs of both VP1 and VP2/3 were disrupted, virus did not bind efficiently to importin β1 and its infectivity remarkably decreased (by 80%). Nuclear targeting of capsid proteins was improved when VP1 and VP2 were co-expressed. VP1 and VP2 were effectively delivered into the nucleus, even when one of the NLS, either VP1 or VP2, was disrupted. Altogether, our results showed that MPyV virions can use VP1 and/or VP2/VP3 NLSs in concert or individually to bind importins to deliver their genomes into the cell nucleus.

Ventral Pallidum Neurons Encode Incentive Value and Promote Cue-Elicited Instrumental Actions.

PubMed

Richard, Jocelyn M; Ambroggi, Frederic; Janak, Patricia H; Fields, Howard L

2016-06-15

The ventral pallidum (VP) is posited to contribute to reward seeking by conveying upstream signals from the nucleus accumbens (NAc). Yet, very little is known about how VP neuron responses contribute to behavioral responses to incentive cues. Here, we recorded activity of VP neurons in a cue-driven reward-seeking task previously shown to require neural activity in the NAc. We find that VP neurons encode both learned cue value and subsequent reward seeking and that activity in VP neurons is required for robust cue-elicited reward seeking. Surprisingly, the onset of VP neuron responses occurs at a shorter latency than cue-elicited responses in NAc neurons. This suggests that this VP encoding is not a passive response to signals generated in the NAc and that VP neurons integrate sensory and motivation-related information received directly from other mesocorticolimbic inputs. Copyright © 2016 Elsevier Inc. All rights reserved.

Mapping the hemagglutination domain of rotaviruses.

PubMed Central

Fuentes-Pananá, E M; López, S; Gorziglia, M; Arias, C F

1995-01-01

Most strains of animal rotaviruses are able to agglutinate erythrocytes, and the surface protein VP4 is the virus hemagglutinin. To map the hemagglutination domain on VP4 while preserving the conformation of the protein, we constructed full-length chimeras between the VP4 genes of hemagglutinating (YM) and nonhemagglutinating (KU) rotavirus strains. The parental and chimeric genes were expressed in insect cells, and the recombinant VP4 proteins were evaluated for their capacity to agglutinate human type O erythrocytes. Three chimeric genes, encoding amino acids 1 to 208 (QKU), 93 to 208 (QC), and 93 to 776 (QYM) of the YM VP4 protein in a KU VP4 background, were constructed. YM VP4 and chimeras QKU and QC were shown to specifically hemagglutinate, indicating that the region between amino acids 93 and 208 of YM VP4 is sufficient to determine the hemagglutination activity of the protein. PMID:7884915

Structure of Rotavirus Outer-Layer Protein VP7 Bound with a Neutralizing Fab

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

Aoki, Scott T.; Settembre, Ethan C.; Trask, Shane D.

2009-06-17

Rotavirus outer-layer protein VP7 is a principal target of protective antibodies. Removal of free calcium ions (Ca{sup 2+}) dissociates VP7 trimers into monomers, releasing VP7 from the virion, and initiates penetration-inducing conformational changes in the other outer-layer protein, VP4. We report the crystal structure at 3.4 angstrom resolution of VP7 bound with the Fab fragment of a neutralizing monoclonal antibody. The Fab binds across the outer surface of the intersubunit contact, which contains two Ca{sup 2+} sites. Mutations that escape neutralization by other antibodies suggest that the same region bears the epitopes of most neutralizing antibodies. The monovalent Fab ismore » sufficient to neutralize infectivity. We propose that neutralizing antibodies against VP7 act by stabilizing the trimer, thereby inhibiting the uncoating trigger for VP4 rearrangement. A disulfide-linked trimer is a potential subunit immunogen.« less

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

Sanchez, R.O.

Voltage breakdowns become a major concern in reducing the size of high-voltage power converter transformers. Even the smallest of voids can provide a path for corona discharge which can cause a dielectric breakdown leading to a transformer failure. A method of encapsulating small high voltage transformers has been developed. The method virtually eliminates voids in the impregnation material, provides an exceptional dielectric between windings and provides a mechanically rugged package. The encapsulation material is a CTBN modified mica filled epoxy. The method requires heat/vacuum to impregnate the coil and heat/pressure to cure the encapsulant. The transformer package utilizes a Diallylmore » Phthalate (DAP) contact assembly in which a coated core/coil assembly is mounted and soldered. This assembly is then loaded into an RTV mold and the encapsulation process begins.« less

Effects of retail style or food service style packaging type and storage time on sensory characteristics of bacon manufactured from commercially sourced pork bellies.

PubMed

Lowe, B K; Bohrer, B M; Holmer, S F; Boler, D D; Dilger, A C

2014-06-01

Objectives were to characterize differences in pork bellies that were stored frozen for different durations prior to processing and characterize sensory properties of the bacon derived from those bellies when stored in either retail or food service style packaging. Bellies (n = 102) were collected from 4 different time periods, fresh bellies (never frozen) and bellies frozen for 2, 5, or 7 mo, and manufactured into bacon under commercial conditions. Food service bacon was packaged in oxygen-permeable polyvinyl lined boxes layered on wax-covered lined paper and blast frozen (-33 °C) for 45 or 90 d after slicing. Retail bacon was vacuum-packaged in retail packages and refrigerated (2 °C) in the dark for 60 or 120 d after slicing. At the end of respective storage times after slicing, bacon was analyzed for sensory attributes and lipid oxidation. Off-flavor and oxidized odor of bacon increased (P < 0.01) with increasing storage time in both packaging types. Lipid oxidation increased (P < 0.01) as storage time increased from day 0 to day 45 in food service packaged bacon from frozen bellies, but was unchanged (P ≥ 0.07) with time in food service packaged bacon from fresh bellies. Lipid oxidation was also unchanged (P ≥ 0.21) over time in retail packaged bacon, with the exception of bellies frozen for 5 mo, which was increased from day 0 to day 90. Overall, off-flavor, oxidized odor, and lipid oxidation increased as storage time after processing increased. Freezing bellies before processing may exacerbate lipid oxidation as storage time after processing was extended. Bacon can be packaged and managed several different ways before it reaches the consumer. This research simulated food service (frozen) and retail packaged (refrigerated) bacon over a range of storage times after slicing. Off-flavor and oxidized odor increased as storage time after processing increased in both packaging types. Lipid oxidation increased as storage time after slicing increased to a greater extent in food service packaging. © 2014 Institute of Food Technologists®

Image quality and radiation dose of lower extremity CT angiography at 70 kVp on an integrated circuit detector dual-source computed tomography.

PubMed

Qi, Li; Zhao, Yan'E; Zhou, Chang Sheng; Spearman, James V; Renker, Matthias; Schoepf, U Joseph; Zhang, Long Jiang; Lu, Guang Ming

2015-06-01

Despite the well-established requirement for radiation dose reduction there are few studies examining the potential for lower extremity CT angiography (CTA) at 70 kVp. To compare the image quality and radiation dose of lower extremity CTA at 70 kVp using a dual-source CT system with an integrated circuit detector to similar studies at 120 kVp. A total of 62 patients underwent lower extremity CTA. Thirty-one patients were examined at 70 kVp using a second generation dual-source CT with an integrated circuit detector (70 kVp group) and 31 patients were evaluated at 120 kVp using a first generation dual-source CT (120 kVp group). The attenuation and image noise were measured and signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) were calculated. Two radiologists assessed image quality. Radiation dose was compared. The mean attenuation of the 70 kVp group was higher than the 120 kVp group (575 ± 149 Hounsfield units [HU] vs. 258 ± 38 HU, respectively, P < 0.001) as was SNR (44.0 ± 22.0 vs 32.7 ± 13.3, respectively, P = 0.017), CNR (39.7 ± 20.6 vs 26.6 ± 11.7, respectively, P = 0.003) and the mean image quality score (3.7 ± 0.1 vs. 3.2 ± 0.3, respectively, P < 0.001). The inter-observer agreement was good for the 70 kVp group and moderate for the 120 kVp group. The dose-length product was lower in the 70 kVp group (264.5 ± 63.1 mGy × cm vs. 412.4 ± 81.5 mGy × cm, P < 0.001). Lower extremity CTA at 70 kVp allows for lower radiation dose with higher SNR, CNR, and image quality when compared with standard 120 kVp. © The Foundation Acta Radiologica 2014 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

In situ structures of the segmented genome and RNA polymerase complex inside a dsRNA virus

NASA Astrophysics Data System (ADS)

Zhang, Xing; Ding, Ke; Yu, Xuekui; Chang, Winston; Sun, Jingchen; Hong Zhou, Z.

2015-11-01

Viruses in the Reoviridae, like the triple-shelled human rotavirus and the single-shelled insect cytoplasmic polyhedrosis virus (CPV), all package a genome of segmented double-stranded RNAs (dsRNAs) inside the viral capsid and carry out endogenous messenger RNA synthesis through a transcriptional enzyme complex (TEC). By direct electron-counting cryoelectron microscopy and asymmetric reconstruction, we have determined the organization of the dsRNA genome inside quiescent CPV (q-CPV) and the in situ atomic structures of TEC within CPV in both quiescent and transcribing (t-CPV) states. We show that the ten segmented dsRNAs in CPV are organized with ten TECs in a specific, non-symmetric manner, with each dsRNA segment attached directly to a TEC. The TEC consists of two extensively interacting subunits: an RNA-dependent RNA polymerase (RdRP) and an NTPase VP4. We find that the bracelet domain of RdRP undergoes marked conformational change when q-CPV is converted to t-CPV, leading to formation of the RNA template entry channel and access to the polymerase active site. An amino-terminal helix from each of two subunits of the capsid shell protein (CSP) interacts with VP4 and RdRP. These findings establish the link between sensing of environmental cues by the external proteins and activation of endogenous RNA transcription by the TEC inside the virus.

Identification of a nuclear localization sequence in the polyomavirus capsid protein VP2

NASA Technical Reports Server (NTRS)

Chang, D.; Haynes, J. I. 2nd; Brady, J. N.; Consigli, R. A.; Spooner, B. S. (Principal Investigator)

1992-01-01

A nuclear localization signal (NLS) has been identified in the C-terminal (Glu307-Glu-Asp-Gly-Pro-Gln-Lys-Lys-Lys-Arg-Arg-Leu318) amino acid sequence of the polyomavirus minor capsid protein VP2. The importance of this amino acid sequence for nuclear transport of newly synthesized VP2 was demonstrated by a genetic "subtractive" study using the constructs pSG5VP2 (expressing full-length VP2) and pSG5 delta 3VP2 (expressing truncated VP2, lacking amino acids Glu307-Leu318). These constructs were transfected into COS-7 cells, and the intracellular localization of the VP2 protein was determined by indirect immunofluorescence. These studies revealed that the full-length VP2 was localized in the nucleus, while the truncated VP2 protein was localized in the cytoplasm and not transported to the nucleus. A biochemical "additive" approach was also used to determine whether this sequence could target nonnuclear proteins to the nucleus. A synthetic peptide identical to VP2 amino acids Glu307-Leu318 was cross-linked to the nonnuclear proteins bovine serum albumin (BSA) or immunoglobulin G (IgG). The conjugates were then labeled with fluorescein isothiocyanate and microinjected into the cytoplasm of NIH 3T6 cells. Both conjugates localized in the nucleus of the microinjected cells, whereas unconjugated BSA and IgG remained in the cytoplasm. Taken together, these genetic subtractive and biochemical additive approaches have identified the C-terminal sequence of polyoma-virus VP2 (containing amino acids Glu307-Leu318) as the NLS of this protein.

VP24 Is a Chitin-Binding Protein Involved in White Spot Syndrome Virus Infection

PubMed Central

Li, Zaipeng; Han, Yali; Xu, Limei

2015-01-01

ABSTRACT Oral ingestion is the major route of infection for the white spot syndrome virus (WSSV). However, the mechanism by which virus particles in the digestive tract invade host cells is unknown. In the present study, we demonstrate that WSSV virions can bind to chitin through one of the major envelope proteins (VP24). Mutagenesis analysis indicated that amino acids (aa) 186 to 200 in the C terminus of VP24 were required for chitin binding. Moreover, the P-VP24186–200 peptide derived from the VP24 chitin binding region significantly inhibited the VP24-chitin interaction and the WSSV-chitin interaction, implying that VP24 participates in WSSV binding to chitin. Oral inoculation experiments showed that P-VP24186–200 treatment reduced the number of virus particles remaining in the digestive tract during the early stage of infection and greatly hindered WSSV proliferation in shrimp. These data indicate that binding of WSSV to chitin through the viral envelope protein VP24 is essential for WSSV per os infection and provide new ideas for preventing WSSV infection in shrimp farms. IMPORTANCE In this study, we show that WSSV can bind to chitin through the envelope protein VP24. The chitin-binding domain of VP24 maps to amino acids 186 to 200 in the C terminus. Binding of WSSV to chitin through the viral envelope protein VP24 is essential for WSSV per os infection. These findings not only extend our knowledge of WSSV infection but also provide new insights into strategies to prevent WSSV infection in shrimp farms. PMID:26512091

Effect of intertidal exposure on Vibrio parahaemolyticus levels in Pacific Northwest oysters.

PubMed

Nordstrom, J L; Kaysner, C A; Blackstone, G M; Vickery, M C L; Bowers, J C; DePaola, A

2004-10-01

Interest in Vibrio parahaemolyticus (Vp) increased in the United States following Vp-associated gastroenteritis outbreaks in 1997 and 1998 involving the West Coast and other areas. The present study evaluated multiple aspects of Vp ecology in the Pacific Northwest with three objectives: (i) to determine the effect of low-tide exposure on Vp levels in oysters, (ii) to determine the relationship between total and pathogenic Vp, and (iii) to examine sediments and aquatic fauna as reservoirs for pathogenic Vp. Samples were collected from intertidal reefs along Hood Canal, Wash., in August 2001. Fecal matter from marine mammals and aquatic birds as well as intestinal contents from bottom-dwelling fish were tested. Total and pathogenic Vp levels in all the samples were enumerated with colony hybridization procedures using DNA probes that targeted the thermolabile direct hemolysin (tlh) and thermostable direct hemolysin (tdh) genes, respectively. The mean Vp densities in oysters were four to eight times greater at maximum exposure than at the corresponding first exposure. While tdh-positive Vp counts were generally < or = 10 CFU/g at first exposure, counts as high as 160 CFU/g were found at maximum exposure. Vp concentrations in sediments were not significantly different from those in oysters at maximum exposure. Pathogenic (tdh positive) Vp was detected in 9 of 42 (21%) oyster samples at maximum exposure, in 5 of 19 (26%) sediment samples, but in 0 of 9 excreta samples. These results demonstrate that summer conditions permit the multiplication of Vp in oysters exposed by a receding tide.

RHETT/EPDM Performance Characterization

NASA Technical Reports Server (NTRS)

Haag, T.; Osborn, M.

1998-01-01

The 0.6 kW Electric Propulsion Demonstration Module (EPDM) flight thruster system was tested in a large vacuum facility for performance measurements and functional checkout. The thruster was operated at a xenon flow rate of 3.01 mg/s, which was supplied through a self-contained propellant system. All power was provided through a flight-packaged power processing unit, which was mounted in vacuum on a cold plate. The thruster was cycled through 34 individual startup and shutdown sequences. Operating periods ranged from 3 to 3600 seconds. The system responded promptly to each command sequence and there were no involuntary shutdowns. Direct thrust measurements indicated that steady state thrust was temperature sensitive, and varied from a high of 41.7 mN at 16 C, to a low of 34.8 mN at 110 C. Short duration thruster firings showed rapid response and good repeatability.

Process for preparing lubricating oil from used waste lubricating oil

DOEpatents

Whisman, Marvin L.; Reynolds, James W.; Goetzinger, John W.; Cotton, Faye O.

1978-01-01

A re-refining process is described by which high-quality finished lubricating oils are prepared from used waste lubricating and crankcase oils. The used oils are stripped of water and low-boiling contaminants by vacuum distillation and then dissolved in a solvent of 1-butanol, 2-propanol and methylethyl ketone, which precipitates a sludge containing most of the solid and liquid contaminants, unspent additives, and oxidation products present in the used oil. After separating the purified oil-solvent mixture from the sludge and recovering the solvent for recycling, the purified oil is preferably fractional vacuum-distilled, forming lubricating oil distillate fractions which are then decolorized and deodorized to prepare blending stocks. The blending stocks are blended to obtain a lubricating oil base of appropriate viscosity before being mixed with an appropriate additive package to form the finished lubricating oil product.

Degradation and Its Control of Ultraviolet Avalanche Photodiodes Using PEDOT:PSS/ZnSSe Organic-Inorganic Hybrid Structure

NASA Astrophysics Data System (ADS)

Abe, Tomoki; Uchida, Shigeto; Tanaka, Keita; Fujisawa, Takanobu; Kasada, Hirofumi; Ando, Koshi; Akaiwa, Kazuaki; Ichino, Kunio

2018-05-01

We investigated device degradation in PEDOT:PSS/ZnSSe organic-inorganic hybrid ultraviolet avalanche photodiodes (UV-APDs). ZnSSe/n-GaAs wafers were grown by molecular beam epitaxy, and PEDOT:PSS window layers were formed by inkjet technique. We observed rapid degradation with APD-mode stress (˜ 30 V) in the N2 (4 N) atmosphere, while we observed no marked change in forward bias current stress and photocurrent stress. In the case of a vacuum condition, we observed no detectable degradation in the dark avalanche current with APD-mode stress. Therefore, the degradation in the PEDOT:PSS/ZnSSe interface under the APD-mode stress was caused by the residual water vapor or oxygen in the N2 atmosphere and could be controlled by vacuum packaging.

Development of High Temperature Type Vacuum Insulation Panel using Soluble Polyimide and Characteristic Evaluation

NASA Astrophysics Data System (ADS)

Araki, Kuninari; Kamoto, Daigorou; Matsuoka, Shin-Ichi

The utilization is expected from the high-insulated characteristic as a tool for energy saving also in the high temperature insulation fields as in vacuum insulation panels (VIP) in the future. For high temperature, the material composition and process of VIP were reviewed, the SUS foil was adopted as packaging material, and soluble polyimide was developed as the thermo compression bonding material for high temperature VIP at 150°C. To lower the glass-transition temperature (Tg) under 200°C, we elaborated the new soluble polyimide using aliphatic diamine copolymer, and controlled Tg to about 176°C. By making from trial VIP and evaluations, it was possible to be maintain high performance concerning the coefficient of thermal conductivity [λ<0.008 W/(m·K) at 150°C].

Development of a rapid and simple immunochromatographic assay to identify Vibrio parahaemolyticus.

PubMed

Sakata, Junko; Kawatsu, Kentaro; Iwasaki, Tadashi; Kumeda, Yuko

2015-09-01

To rapidly and simply determine whether or not bacterial colonies growing on agar were Vibrio parahaemolyticus, we developed an immunochromatographic assay (VP-ICA) using two different monoclonal antibodies (designated mAb-VP34 and mAb-VP109) against the delta subunit of V. parahaemolyticus-F0F1 ATP synthase. The epitopes recognized by mAb-VP34 and mAb-VP109 were mapped to sequences of eight ((47)LLTSSFSA(54)) and six amino acid residues ((16)FDFAVD(21)), respectively. An amino acid sequence similarity search of the NCBI database using BLASTP showed that both epitopic amino acid sequences were present together only in V. parahaemolyticus. When 124 V. parahaemolyticus strains and 94 strains of 27 other Vibrio species or 35 non-Vibrio species were tested using the VP-ICA, the VP-ICA identified V. parahaemolyticus with 100% accuracy. The VP-ICA rapidly and simply identified the pathogen directly from a single agar colony within 30 min, indicating that VP-ICA will greatly reduce labor and time required to identify V. parahaemolyticus compared with conventional biochemical tests. Copyright © 2015. Published by Elsevier B.V.

Intrinsically-disordered N-termini in human parechovirus 1 capsid proteins bind encapsidated RNA.

PubMed

Shakeel, Shabih; Evans, James D; Hazelbaker, Mark; Kao, C Cheng; Vaughan, Robert C; Butcher, Sarah J

2018-04-11

Human parechoviruses (HPeV) are picornaviruses with a highly-ordered RNA genome contained within icosahedrally-symmetric capsids. Ordered RNA structures have recently been shown to interact with capsid proteins VP1 and VP3 and facilitate virus assembly in HPeV1. Using an assay that combines reversible cross-linking, RNA affinity purification and peptide mass fingerprinting (RCAP), we mapped the RNA-interacting regions of the capsid proteins from the whole HPeV1 virion in solution. The intrinsically-disordered N-termini of capsid proteins VP1 and VP3, and unexpectedly, VP0, were identified to interact with RNA. Comparing these results to those obtained using recombinantly-expressed VP0 and VP1 confirmed the virion binding regions, and revealed unique RNA binding regions in the isolated VP0 not previously observed in the crystal structure of HPeV1. We used RNA fluorescence anisotropy to confirm the RNA-binding competency of each of the capsid proteins' N-termini. These findings suggests that dynamic interactions between the viral RNA and the capsid proteins modulate virus assembly, and suggest a novel role for VP0.

Radiation dose and image conspicuity comparison between conventional 120 kVp and 150 kVp with spectral beam shaping for temporal bone CT.

PubMed

Kim, Chang Rae; Jeon, Ji Young

2018-05-01

The purpose of this article is to compare radiation doses and conspicuity of anatomic landmarks of the temporal bone between the CT technique using spectral beam shaping at 150 kVp with a dedicated tin filter (150 kVp-Sn) and the conventional protocol at 120 kVp. 25 patients (mean age, 46.8 ± 21.2 years) were examined using the 150-kVp Sn protocol (200 reference mAs using automated tube current modulation, 64 × 0.6 mm collimation, 0.6 mm slice thickness, pitch 0.8), whereas 30 patients (mean age, 54.5 ± 17.8 years) underwent the 120-kVp protocol (180 mAs, 128 × 0.6 mm collimation, 0.6 mm slice thickness, pitch 0.8). Radiation doses were compared between the two acquisition techniques, and dosimetric data from the literature were reviewed for comparison of radiation dose reduction. Subjective conspicuity of 23 anatomic landmarks of the temporal bone, expressed by 5-point rating scale and objective conspicuity by signal-to-noise ratio (SNR) which measured in 4 different regions of interest (ROI), were compared between 150-kVp Sn and 120-kVp acquisitions. The mean dose-length-product (DLP) and effective dose were significantly lower for the 150-kVp Sn scans (0.26 ± 0.26 mSv) compared with the 120-kVp scans (0.92 ± 0.10 mSv, p < 0.001). The lowest effective dose from the literature-based protocols was 0.31 ± 0.12 mSv, which proposed as a low-dose protocol in the setting of spiral multislice temporal bone CT. SNR was slightly superior for 120-kVp images, however analyzability of the 23 anatomic structures did not differ significantly between 150-kVp Sn and 120-kVp scans. Temporal bone CT performed at 150 kVp with an additional tin filter for spectral shaping markedly reduced radiation exposure when compared with conventional temporal bone CT at 120 kVp while maintaining anatomic conspicuity. The decreased radiation dose of the 150-kVp Sn was also lower in comparison to the previous literature-based low-dose temporal bone CT protocol. Copyright © 2018 Elsevier B.V. All rights reserved.

The Ebola Virus VP30-NP Interaction Is a Regulator of Viral RNA Synthesis

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

Kirchdoerfer, Robert N.; Moyer, Crystal L.; Abelson, Dafna M.

Filoviruses are capable of causing deadly hemorrhagic fevers. All nonsegmented negative-sense RNA-virus nucleocapsids are composed of a nucleoprotein (NP), a phosphoprotein (VP35) and a polymerase (L). However, the VP30 RNA-synthesis co-factor is unique to the filoviruses. The assembly, structure, and function of the filovirus RNA replication complex remain unclear. Here, we have characterized the interactions of Ebola, Sudan and Marburg virus VP30 with NP using in vitro biochemistry, structural biology and cell-based mini-replicon assays. We have found that the VP30 C-terminal domain interacts with a short peptide in the C-terminal region of NP. Further, we have solved crystal structures ofmore » the VP30-NP complex for both Ebola and Marburg viruses. These structures reveal that a conserved, proline-rich NP peptide binds a shallow hydrophobic cleft on the VP30 C-terminal domain. Structure-guided Ebola virus VP30 mutants have altered affinities for the NP peptide. Correlation of these VP30-NP affinities with the activity for each of these mutants in a cell-based mini-replicon assay suggests that the VP30-NP interaction plays both essential and inhibitory roles in Ebola virus RNA synthesis.« less

Poly(vinylidene difluoride)/poly(tetrafluoroethylene-co-vinylpyrrolidone) blend membranes with antifouling properties.

PubMed

Sun, Yuchen; Rajabzadeh, Saeid; Fang, Lifeng; Jeon, Sungil; Zhou, Zhuang; Ohmukai, Yoshikage; Miki, Jun; Wang, Xiaolin; Matsuyama, Hideto

2017-06-01

To inhibit fouling phenomenon in membrane process, a new amphiphilic copolymer, poly(tetrafluoroethylene-co-vinylpyrrolidone) (P(TFE-VP)), was blended with poly(vinylidene difluoride) (PVDF) to fabricate a series of antifouling membranes via non solvent induced phase separation (NIPS) method. The effect of copolymer blend ratios and TFE/VP ratios on membrane properties were evaluated, and the stability of P(TFE-VP) in PVDF membrane was studied. The membrane morphology was controlled by adjusting polymer concentration in dope solution, such that all membranes have similar pore size and density, as well as pure water permeability. In evaluating the effect of TFE/VP ratios, the content of VP in dope solutions was also adjusted to allow a fair comparison. We found that for P(TFE-VP) with a higher VP content, adsorption of BSA on polymer film was negligible. Higher blend ratios of this copolymer resulted in higher surface VP content and better hydrophilicity, but antifouling performance ceased to improve when blend ratio was larger than 1:9 (copolymer:PVDF). Meanwhile, a lower VP content in copolymer resulted in inferior hydrophilicity and severe fouling of the blend membranes. It was also proved that comparing with PVP homopolymer, P(TFE-VP) had satisfying stability inside PVDF membrane. Copyright © 2017 Elsevier B.V. All rights reserved.

Effect of bladder augmentation on VP shunt failure rates in spina bifida.

PubMed

Gonzalez, Dani O; Cooper, Jennifer N; McLeod, Daryl J

2017-12-11

Most patients with spina bifida require ventriculoperitoneal (VP) shunt placement. Some also require bladder augmentation, which may increase the risk of VP shunt malfunction and/or failure. The aim of this study was to assess whether bladder augmentation affects the rate of VP shunt failure in this population. Using the Pediatric Health Information System, we studied patients with spina bifida born between 1992 and 2014 who underwent VP shunt placement. Using conditional logistic regression, we compared age- and hospital-matched patients who did and did not undergo a bladder augmentation to determine their difference in rates of VP shunt failure. There were 4192 patients with spina bifida who underwent both surgical closure and VP shunt placement. Of these, 203 patients with bladder augmentation could be matched to 593 patients without bladder augmentation. VP shunt failure occurred within 2 years in 7.7% of patients, the majority of whom were in the group who underwent bladder augmentation (87%). After adjusting for confounders, undergoing bladder augmentation was independently associated with VP shunt failure (HR: 33.5, 95% CI: 13.15-85.44, p< 0.001). Bladder augmentation appears to be associated with VP shunt failure. Additional studies are necessary to better define this relationship and identify risk-reduction techniques.

Conserved differences in protein sequence determine the human pathogenicity of Ebolaviruses

PubMed Central

Pappalardo, Morena; Juliá, Miguel; Howard, Mark J.; Rossman, Jeremy S.; Michaelis, Martin; Wass, Mark N.

2016-01-01

Reston viruses are the only Ebolaviruses that are not pathogenic in humans. We analyzed 196 Ebolavirus genomes and identified specificity determining positions (SDPs) in all nine Ebolavirus proteins that distinguish Reston viruses from the four human pathogenic Ebolaviruses. A subset of these SDPs will explain the differences in human pathogenicity between Reston and the other four ebolavirus species. Structural analysis was performed to identify those SDPs that are likely to have a functional effect. This analysis revealed novel functional insights in particular for Ebolavirus proteins VP40 and VP24. The VP40 SDP P85T interferes with VP40 function by altering octamer formation. The VP40 SDP Q245P affects the structure and hydrophobic core of the protein and consequently protein function. Three VP24 SDPs (T131S, M136L, Q139R) are likely to impair VP24 binding to human karyopherin alpha5 (KPNA5) and therefore inhibition of interferon signaling. Since VP24 is critical for Ebolavirus adaptation to novel hosts, and only a few SDPs distinguish Reston virus VP24 from VP24 of other Ebolaviruses, human pathogenic Reston viruses may emerge. This is of concern since Reston viruses circulate in domestic pigs and can infect humans, possibly via airborne transmission. PMID:27009368

Conserved differences in protein sequence determine the human pathogenicity of Ebolaviruses.

PubMed

Pappalardo, Morena; Juliá, Miguel; Howard, Mark J; Rossman, Jeremy S; Michaelis, Martin; Wass, Mark N

2016-03-24

Reston viruses are the only Ebolaviruses that are not pathogenic in humans. We analyzed 196 Ebolavirus genomes and identified specificity determining positions (SDPs) in all nine Ebolavirus proteins that distinguish Reston viruses from the four human pathogenic Ebolaviruses. A subset of these SDPs will explain the differences in human pathogenicity between Reston and the other four ebolavirus species. Structural analysis was performed to identify those SDPs that are likely to have a functional effect. This analysis revealed novel functional insights in particular for Ebolavirus proteins VP40 and VP24. The VP40 SDP P85T interferes with VP40 function by altering octamer formation. The VP40 SDP Q245P affects the structure and hydrophobic core of the protein and consequently protein function. Three VP24 SDPs (T131S, M136L, Q139R) are likely to impair VP24 binding to human karyopherin alpha5 (KPNA5) and therefore inhibition of interferon signaling. Since VP24 is critical for Ebolavirus adaptation to novel hosts, and only a few SDPs distinguish Reston virus VP24 from VP24 of other Ebolaviruses, human pathogenic Reston viruses may emerge. This is of concern since Reston viruses circulate in domestic pigs and can infect humans, possibly via airborne transmission.

Space Suit Portable Life Support System (PLSS) 2.0 Unmanned Vacuum Environment Testing

NASA Technical Reports Server (NTRS)

Watts, Carly; Vogel, Matthew

2016-01-01

For the first time in more than 30 years, an advanced space suit Portable Life Support System (PLSS) design was operated inside a vacuum chamber representative of the flight operating environment. The test article, PLSS 2.0, was the second system-level integrated prototype of the advanced PLSS design, following the PLSS 1.0 Breadboard that was developed and tested throughout 2011. Whereas PLSS 1.0 included five technology development components with the balance the system simulated using commercial-off-the-shelf items, PLSS 2.0 featured first generation or later prototypes for all components less instrumentation, tubing and fittings. Developed throughout 2012, PLSS 2.0 was the first attempt to package the system into a flight-like representative volume. PLSS 2.0 testing included an extensive functional evaluation known as Pre-Installation Acceptance (PIA) testing, Human-in-the-Loop testing in which the PLSS 2.0 prototype was integrated via umbilicals to a manned prototype space suit for 19 two-hour simulated EVAs, and unmanned vacuum environment testing. Unmanned vacuum environment testing took place from 1/9/15-7/9/15 with PLSS 2.0 located inside a vacuum chamber. Test sequences included performance mapping of several components, carbon dioxide removal evaluations at simulated intravehicular activity (IVA) conditions, a regulator pressure schedule assessment, and culminated with 25 simulated extravehicular activities (EVAs). During the unmanned vacuum environment test series, PLSS 2.0 accumulated 378 hours of integrated testing including 291 hours of operation in a vacuum environment and 199 hours of simulated EVA time. The PLSS prototype performed nominally throughout the test series, with two notable exceptions including a pump failure and a Spacesuit Water Membrane Evaporator (SWME) leak, for which post-test failure investigations were performed. In addition to generating an extensive database of PLSS 2.0 performance data, achievements included requirements and operational concepts verification, as well as demonstration of vehicular interfaces, consumables sizing and recharge, and water quality control.

The retail display life of steaks prepared from chill stored vacuum and carbon dioxide-packed sub-primal beef cuts.

PubMed

Bell, R G; Penney, N; Moorhead, S M

1996-01-01

Chilled striploins and cube rolls from ten Australian steers (grain-fed for 150 days) were trimmed of external fat and cut transversely into portions approximately 10 cm thick, each weighing between 750 and 1000 g. These 'retailer-ready' cuts were each wrapped in drip saver pads and slid inside a plastic sleeve before being individually placed into a clear plastic high oxygen barrier film, metallized film or conventional vacuum bag. Cuts in clear plastic and metallized film packs were packaged in an oxygen-free saturated carbon dioxide atmosphere (CO(2)-CAP), those in vacuum bags were conventionally vacuum-packed. All packs were returned to the chiller for further cooling. After 24 hr, half the clear plastic and metallized CO(2)-CAP packs were carbon dioxide master-packed in groups of eight. Retailer-ready cuts in both clear plastic and metallized film single unit and master-packed CO(2)-CAP packs were air freighted to New Zealand and sea freighted to Japan for assessment. The control vacuum packs were all consigned to New Zealand. Assessments in both countries after 39-89 days storage at between 0 °C and -1.0 °C indicated that fat colour stability limited the retail display life of steaks cut from meat in these retailer-ready packs to approximately 48 hr. In this regard, meat from single unit CO(2)-CAP, master pack CO(2)-CAP and vacuum packs performed similarly. Lean meat colour and sensory attributes remained acceptable for up to 48 hr after displayed product was rejected because of grey-green fat discoloration. The microbiological status of retailer-ready cuts removed from CO(2)-CAP packs after 89 days chilled storage was superior to that of cuts from vacuum packs. Clear plastic and metallized film CO(2)-CAP packs performed comparably.

The prevalence and some metabolic traits of Brochothrix thermosphacta in meat and meat products packaged in different ways.

PubMed

Nowak, Agnieszka; Rygala, Anna; Oltuszak-Walczak, Elzbieta; Walczak, Piotr

2012-04-01

The effect of Brochothrix thermosphacta on the quality of meat and meat products is of vital importance in connection with Regulation EC/178/2002 extending the definition of unsafe foodstuffs to encompass all those which are unfit for human consumption. This study aimed to determine the prevalence of B. thermosphacta in meat and meat products packaged under different conditions and to estimate the effect of B. thermosphacta strains on product quality based on their protein and lipid degradation activity. B. thermosphacta was absent in only two of 132 samples. All other samples were contaminated with this bacterium (10(1) to 10(9) cfu g(-1) meat and 10(2) to 10(8) cfu g(-1) meat product). In products stored under high-oxygen atmosphere Brochothrix cells accounted for almost 100% total mesophilic count (TMC) and below 50% TMC in oxygen-free atmosphere. While the tested B. thermosphacta strains did not show any proteolytic activity, most of them displayed lipolytic activity at 25 °C and some even at 4 °C. B. thermosphacta is commonly present in meat and meat products packaged in different ways. This bacterium can display lipolytic activity also at refrigeration temperature. Its over-proliferation can be inhibited through vacuum packaging or packaging under a modified atmosphere with reduced oxygen content. Copyright © 2011 Society of Chemical Industry.

Molecular cloning, expression and first antigenic characterization of human astrovirus VP26 structural protein and a C-terminal deleted form.

PubMed

Royuela, Enrique; Sánchez-Fauquier, Alicia

2010-01-01

The open reading frame 2 (ORF2) of human astrovirus (HAstV) encodes the structural VP26 protein that seems to be the main antigenic viral protein. However, its functional role remains unclear. Bioinformatic predictions revealed that VP29 and VP26 proteins could be involved in virus-cell interaction. In this study, we describe for the first time the cloning and expression in Escherichia coli (E. coli) of a recombinant VP26 (rVP26) protein and a VP26 C-terminal truncated form (VP26 Delta C), followed by purification by NTA-Ni(2+) agarose affinity chromatography. Protein expression and purification were evaluated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot (WB). Then, the purified proteins were evaluated for antigenic properties in enzyme linked immunosorbent assay (ELISA) using a polyclonal antibody (PAb) and a neutralizing monoclonal antibody (nMAb) named PL2, both of them directed to HAstV. The results presented herein indicate that the C-terminal end of the VP26 protein is essential to maintain the neutralizing epitope recognized by nMAb PL2 and that the N-terminus of VP26 protein may contain antigenic lineal-epitopes recognized by PAb. Thus, these recombinant proteins can be ideal tools for further antigenic, biochemical, structural and functional VP26 protein characterization, in order to evaluate its potential role in immunodiagnosis and vaccine studies.

Production of rotavirus-like particles in tomato (Lycopersicon esculentum L.) fruit by expression of capsid proteins VP2 and VP6 and immunological studies.

PubMed

Saldaña, Sergio; Esquivel Guadarrama, Fernando; Olivera Flores, Teresa De Jesús; Arias, Nancy; López, Susana; Arias, Carlos; Ruiz-Medrano, Roberto; Mason, Hugh; Mor, Tsafrir; Richter, Liz; Arntzen, Charles J; Gómez Lim, Miguel A

2006-01-01

A number of different antigens have been successfully expressed in transgenic plants, and some are currently being evaluated as orally delivered vaccines. Here we report the successful expression of rotavirus capsid proteins VP2 and VP6 in fruits of transgenic tomato plants. By western blot analysis, using specific antibodies, we determined that the VP2 and VP6 produced in plants have molecular weights similar to those found in native rotavirus. The plant-synthesized VP6 protein retained the capacity to form trimers. We were able to recover rotavirus virus-like particles from tomato fruit (i.e., tomatoes) by centrifugation on a sucrose cushion and to visualize them by electron microscopy. This result indicated that VP2/VP6 can self-assemble into virus-like particles (VLPs) in plant cells, even though only a small proportion of VP2/VP6 assembled into VLPs. To investigate immunogenicity, adult mice were immunized intraperitoneally (i.p.) three times with a protein extract from a transgenic tomatoes in adjuvant. We found that the transgenic tomato extract induced detectable levels of anti-rotavirus antibodies in serum; however, we did not determine the contribution of either the free rotavirus proteins or the VLPs to the induction of the antibody response. These results suggest the potential of plant-based rotavirus VLPs for the development of a vaccine against rotavirus infection.

Mutations Abrogating VP35 Interaction with Double-Stranded RNA Render Ebola Virus Avirulent in Guinea Pigs

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

Prins, Kathleen C.; Delpeut, Sebastien; Leung, Daisy W.

2010-10-11

Ebola virus (EBOV) protein VP35 is a double-stranded RNA (dsRNA) binding inhibitor of host interferon (IFN)-{alpha}/{beta} responses that also functions as a viral polymerase cofactor. Recent structural studies identified key features, including a central basic patch, required for VP35 dsRNA binding activity. To address the functional significance of these VP35 structural features for EBOV replication and pathogenesis, two point mutations, K319A/R322A, that abrogate VP35 dsRNA binding activity and severely impair its suppression of IFN-{alpha}/{beta} production were identified. Solution nuclear magnetic resonance (NMR) spectroscopy and X-ray crystallography reveal minimal structural perturbations in the K319A/R322A VP35 double mutant and suggest that lossmore » of basic charge leads to altered function. Recombinant EBOVs encoding the mutant VP35 exhibit, relative to wild-type VP35 viruses, minimal growth attenuation in IFN-defective Vero cells but severe impairment in IFN-competent cells. In guinea pigs, the VP35 mutant virus revealed a complete loss of virulence. Strikingly, the VP35 mutant virus effectively immunized animals against subsequent wild-type EBOV challenge. These in vivo studies, using recombinant EBOV viruses, combined with the accompanying biochemical and structural analyses directly correlate VP35 dsRNA binding and IFN inhibition functions with viral pathogenesis. Moreover, these studies provide a framework for the development of antivirals targeting this critical EBOV virulence factor.« less

Temporal expression and immunogold localization of Plodia interpunctella granulosis virus structural proteins

NASA Technical Reports Server (NTRS)

Funk, C. J.; Consigli, R. A.; Spooner, B. S. (Principal Investigator)

1993-01-01

Monospecific antisera were produced against four structural proteins (VP12, VP17, VP31, and granulin) of the Plodia interpunctella granulosis virus using polypeptides derived by sodium dodecyl sulfate-polyacrylamide gel electrophoresis or acid extraction. The antisera were shown to be specific on immunoblots of SDS-PAGE separated granulosis virus and were further used to detect structural proteins in infected fat body lysates. Immunoblots of fat body lysates from early stages of infection indicated that VP12, VP17, VP31, and granulin were expressed by 2.5 days post-infection. Immunogold labeling of the virus using the monospecific antisera and electron microscopy confirmed earlier reports that granulin is located in the protein matrix, V17 is an envelope protein, and VP31 is a capsid protein.

Effect of meat ingredients (sodium nitrite and erythorbate) and processing (vacuum storage and packaging atmosphere) on germination and outgrowth of Clostridium perfringens spores in ham during abusive cooling

USDA-ARS?s Scientific Manuscript database

The effect of nitrite and erythorbate on Clostridium perfringens spore germination and outgrowth in ham during abusive cooling (15 h) was evaluated. Ham was formulated with ground pork, NaNO2 (0, 50, 100, 150 or 200 ppm) and sodium erythorbate (0 or 547 ppm). Ten grams of meat (stored at 5C for 3 or...

Multifunctional Space Evaporator-Absorber-Radiator (SEAR)

NASA Technical Reports Server (NTRS)

Bue, Grant C.; Hodgson, Ed; Izenson, Mike; Chen, Weibo

2013-01-01

A system for non-venting thermal control for spacesuits was built by integrating two previously developed technologies, namely NASA's Spacesuit Water Membrane Evaporator (SWME), and Creare's flexible version of the Lithium Chloride Absorber Radiator (LCAR). This SEAR system was tested in relevant thermal vacuum conditions. These tests show that a 1 sq m radiator having about three times as much absorption media as in the test article would be required to support a 7 hour spacewalk. The serial flow arrangement of the LCAR of the flexible version proved to be inefficient for venting non-condensable gas (NCG). A different LCAR packaging arrangement was conceived wherein the Portable Life Support System (PLSS) housing would be made with a high-strength carbon fiber composite honeycomb, the cells of which would be filled with the chemical absorption media. This new packaging reduce the mass and volume impact of the SEAR on the Portable Life Support System (PLSS) compared to the flexible design. A 0.2 sq m panel with flight-like honeycomb geometry is being constructed and will be tested in thermal and thermal vacuum conditions. Design analyses forecast improved system performance and improved NCG control. A flight-like regeneration system also is also being built and tested. Design analyses for the structurally integrated prototype as well as the earlier test data show that SEAR is not only practical for spacesuits but also has useful applications in spacecraft thermal control.

Bruises in beef cattle at slaughter in Mexico: implications on quality, safety and shelf life of the meat.

PubMed

Cruz-Monterrosa, Rosy G; Reséndiz-Cruz, Verónica; Rayas-Amor, Armando A; López, Marcos; la Lama, Genaro C Miranda-de

2017-01-01

In emergent economies and developing countries of Africa, Asia and Latin America, the major cause for carcass rejection from the international market is bruising; nevertheless, many of these carcases are destined to local markets and meat processing industries for human consumption. Therefore, the aim of the present study was to assess the effect of bruised meat on pH, microbiologic count and biogenic amine (BA) profiles along 21 days of ageing (sampling 1st, 7th, 14th and 21st day) with two packaging method (plastic bag vs vacuum) at 4 °C. A total of 50 bruised carcasses were sampled from 1000 young bulls (Brown Swiss X Zebu) of 18-24 months old and an average live weight of 450 ± 66 kg. The results showed significant differences between packaging systems and bruised vs non-bruised meat. The bruised meat caused higher biogenic amine concentrations than did non-bruised meat. We conclude that bruised meat favoured increments of biogenic amine concentrations, even more than did non-bruised meat. The plastic bag + vacuum system limited the increments of BA concentration during storage time therefore it improved shelf life of meat. These results emphasized the importance of implementing best management practices during pre-slaughter operations of cattle in order to reduce a possible risk factor for bruised meat.

Space Evaporator-Absorber-Radiator (SEAR)

NASA Technical Reports Server (NTRS)

Bue, Grant C.; Stephan, Ryan; Hodgson, Ed; Izenson, Mike; Chen, Weibo

2012-01-01

A system for non-venting thermal control for spacesuits was built by integrating two previously developed technologies, namely NASA s Spacesuit Water Membrane Evaporator (SWME), and Creare s flexible version of the Lithium Chloride Absorber Radiator (LCAR). This SEAR system was tested in relevant thermal vacuum conditions. These tests show that a 1 m2 radiator having about three times as much absorption media as in the test article would be required to support a 7 hour spacewalk. The serial flow arrangement of the LCAR of the flexible version proved to be inefficient for venting non-condensable gas (NCG). A different LCAR packaging arrangement was conceived wherein the Portable Life Support System (PLSS) housing would be made with a high-strength carbon fiber composite honeycomb, the cells of which would be filled with the chemical absorption media. This new packaging reduces the mass and volume impact of the SEAR on the Portable Life Support System (PLSS) compared to the flexible design. A 0.2 sq m panel with flight-like honeycomb geometry is being constructed and will be tested in thermal and thermal vacuum conditions. Design analyses forecast improved system performance and improved NCG control. A flight-like regeneration system also is also being built and tested. Design analyses for the structurally integrated prototype as well as the earlier test data show that SEAR is not only practical for spacesuits but also has useful applications in spacecraft thermal control.

C Terminus of Infectious Bursal Disease Virus Major Capsid Protein VP2 Is Involved in Definition of the T Number for Capsid Assembly

PubMed Central

Castón, José R.; Martínez-Torrecuadrada, Jorge L.; Maraver, Antonio; Lombardo, Eleuterio; Rodríguez, José F.; Casal, J. Ignacio; Carrascosa, José L.

2001-01-01

Infectious bursal disease virus (IBDV), a member of the Birnaviridae family, is a double-stranded RNA virus. The IBDV capsid is formed by two major structural proteins, VP2 and VP3, which assemble to form a T=13 markedly nonspherical capsid. During viral infection, VP2 is initially synthesized as a precursor, called VPX, whose C end is proteolytically processed to the mature form during capsid assembly. We have computed three-dimensional maps of IBDV capsid and virus-like particles built up by VP2 alone by using electron cryomicroscopy and image-processing techniques. The IBDV single-shelled capsid is characterized by the presence of 260 protruding trimers on the outer surface. Five classes of trimers can be distinguished according to their different local environments. When VP2 is expressed alone in insect cells, dodecahedral particles form spontaneously; these may be assembled into larger, fragile icosahedral capsids built up by 12 dodecahedral capsids. Each dodecahedral capsid is an empty T=1 shell composed of 20 trimeric clusters of VP2. Structural comparison between IBDV capsids and capsids consisting of VP2 alone allowed the determination of the major capsid protein locations and the interactions between them. Whereas VP2 forms the outer protruding trimers, VP3 is found as trimers on the inner surface and may be responsible for stabilizing functions. Since elimination of the C-terminal region of VPX is correlated with the assembly of T=1 capsids, this domain might be involved (either alone or in cooperation with VP3) in the induction of different conformations of VP2 during capsid morphogenesis. PMID:11602723

Phylogenetic characterization of Canine Parvovirus VP2 partial sequences from symptomatic dogs samples.

PubMed

Zienius, D; Lelešius, R; Kavaliauskis, H; Stankevičius, A; Šalomskas, A

2016-01-01

The aim of the present study was to detect canine parvovirus (CPV) from faecal samples of clinically ill domestic dogs by polymerase chain reaction (PCR) followed by VP2 gene partial sequencing and molecular characterization of circulating strains in Lithuania. Eleven clinically and antigen-tested positive dog faecal samples, collected during the period of 2014-2015, were investigated by using PCR. The phylogenetic investigations indicated that the Lithuanian CPV VP2 partial sequences (3025-3706 cds) were closely related and showed 99.0-99.9% identity. All Lithuanian sequences were associated with one phylogroup, but grouped in different clusters. Ten of investigated Lithuanian CPV VP2 sequences were closely associated with CPV 2a antigenic variant (99.4% nt identity). Five CPV VP2 sequences from Lithuania were related to CPV-2a, but were rather divergent (6.8 nt differences). Only one CPV VP2 sequence from Lithuania was associated (99.3% nt identity) with CPV-2b VP2 sequences from France, Italy, USA and Korea. The four of eleven investigated Lithuanian dogs with CPV infection symptoms were vaccinated with CPV-2 vaccine, but their VP2 sequences were phylogenetically distantly associated with CPV vaccine strains VP2 sequences (11.5-15.8 nt differences). Ten Lithuanian CPV VP2 sequences had monophyletic relations among the close geographically associated samples, but five of them were rather divergent (1.0% less sequence similarity). The one Lithuanian CPV VP2 sequence was closely related with CPV-2b antigenic variant. All the Lithuanian CPV VP2 partial sequences were conservative and phylogenetically low associated with most commonly used CPV vaccine strains.

Expression, purification and crystallization of two major envelope proteins from white spot syndrome virus

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

Tang, Xuhua; Hew, Choy Leong, E-mail: dbshewcl@nus.edu.sg

2007-07-01

The crystallization of the N-terminal transmembrane region-truncated VP26 and VP28 of white spot syndrome virus is described. White spot syndrome virus (WSSV) is a major virulent pathogen known to infect penaeid shrimp and other crustaceans. VP26 and VP28, two major envelope proteins from WSSV, have been identified and overexpressed in Escherichia coli. In order to facilitate purification and crystallization, predicted N-terminal transmembrane regions of approximately 35 amino acids have been truncated from both VP26 and VP28. Truncated VP26 and VP28 and their corresponding SeMet-labelled proteins were purified and the SeMet proteins were crystallized by the hanging-drop vapour-diffusion method. Crystals ofmore » SeMet-labelled VP26 were obtained using a reservoir consisting of 0.1 M citric acid pH 3.5, 3.0 M sodium chloride and 1%(w/v) polyethylene glycol 3350, whereas SeMet VP28 was crystallized using a reservoir solution consisting of 25% polyethylene glycol 8000, 0.2 M calcium acetate, 0.1 M Na HEPES pH 7.5 and 1.5%(w/v) 1,2,3-heptanetriol. Crystals of SeMet-labelled VP26 diffract to 2.2 Å resolution and belong to space group R32, with unit-cell parameters a = b = 73.92, c = 199.31 Å. SeMet-labelled VP28 crystallizes in space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 105.33, b = 106.71, c = 200.37 Å, and diffracts to 2.0 Å resolution.« less

RNA binding specificity of Ebola virus transcription factor VP30.

PubMed

Schlereth, Julia; Grünweller, Arnold; Biedenkopf, Nadine; Becker, Stephan; Hartmann, Roland K

2016-09-01

The transcription factor VP30 of the non-segmented RNA negative strand Ebola virus balances viral transcription and replication. Here, we comprehensively studied RNA binding by VP30. Using a novel VP30:RNA electrophoretic mobility shift assay, we tested truncated variants of 2 potential natural RNA substrates of VP30 - the genomic Ebola viral 3'-leader region and its complementary antigenomic counterpart (each ∼155 nt in length) - and a series of other non-viral RNAs. Based on oligonucleotide interference, the major VP30 binding region on the genomic 3'-leader substrate was assigned to the internal expanded single-stranded region (∼ nt 125-80). Best binding to VP30 was obtained with ssRNAs of optimally ∼ 40 nt and mixed base composition; underrepresentation of purines or pyrimidines was tolerated, but homopolymeric sequences impaired binding. A stem-loop structure, particularly at the 3'-end or positioned internally, supports stable binding to VP30. In contrast, dsRNA or RNAs exposing large internal loops flanked by entirely helical arms on both sides are not bound. Introduction of a 5´-Cap(0) structure impaired VP30 binding. Also, ssDNAs bind substantially weaker than isosequential ssRNAs and heparin competes with RNA for binding to VP30, indicating that ribose 2'-hydroxyls and electrostatic contacts of the phosphate groups contribute to the formation of VP30:RNA complexes. Our results indicate a rather relaxed RNA binding specificity of filoviral VP30, which largely differs from that of the functionally related transcription factor of the Paramyxoviridae which binds to ssRNAs as short as 13 nt with a preference for oligo(A) sequences.

Detection of lipid-induced structural changes of the Marburg virus matrix protein VP40 using hydrogen/deuterium exchange-mass spectrometry

PubMed Central

Wijesinghe, Kaveesha J.; Urata, Sarah; Bhattarai, Nisha; Kooijman, Edgar E.; Gerstman, Bernard S.; Chapagain, Prem P.; Li, Sheng; Stahelin, Robert V.

2017-01-01

Marburg virus (MARV) is a lipid-enveloped virus from the Filoviridae family containing a negative sense RNA genome. One of the seven MARV genes encodes the matrix protein VP40, which forms a matrix layer beneath the plasma membrane inner leaflet to facilitate budding from the host cell. MARV VP40 (mVP40) has been shown to be a dimeric peripheral protein with a broad and flat basic surface that can associate with anionic phospholipids such as phosphatidylserine. Although a number of mVP40 cationic residues have been shown to facilitate binding to membranes containing anionic lipids, much less is known on how mVP40 assembles to form the matrix layer following membrane binding. Here we have used hydrogen/deuterium exchange (HDX) mass spectrometry to determine the solvent accessibility of mVP40 residues in the absence and presence of phosphatidylserine and phosphatidylinositol 4,5-bisphosphate. HDX analysis demonstrates that two basic loops in the mVP40 C-terminal domain make important contributions to anionic membrane binding and also reveals a potential oligomerization interface in the C-terminal domain as well as a conserved oligomerization interface in the mVP40 N-terminal domain. Lipid binding assays confirm the role of the two basic patches elucidated with HD/X measurements, whereas molecular dynamics simulations and membrane insertion measurements complement these studies to demonstrate that mVP40 does not appreciably insert into the hydrocarbon region of anionic membranes in contrast to the matrix protein from Ebola virus. Taken together, we propose a model by which association of the mVP40 dimer with the anionic plasma membrane facilitates assembly of mVP40 oligomers. PMID:28167534

Detection of lipid-induced structural changes of the Marburg virus matrix protein VP40 using hydrogen/deuterium exchange-mass spectrometry.

PubMed

Wijesinghe, Kaveesha J; Urata, Sarah; Bhattarai, Nisha; Kooijman, Edgar E; Gerstman, Bernard S; Chapagain, Prem P; Li, Sheng; Stahelin, Robert V

2017-04-14

Marburg virus (MARV) is a lipid-enveloped virus from the Filoviridae family containing a negative sense RNA genome. One of the seven MARV genes encodes the matrix protein VP40, which forms a matrix layer beneath the plasma membrane inner leaflet to facilitate budding from the host cell. MARV VP40 (mVP40) has been shown to be a dimeric peripheral protein with a broad and flat basic surface that can associate with anionic phospholipids such as phosphatidylserine. Although a number of mVP40 cationic residues have been shown to facilitate binding to membranes containing anionic lipids, much less is known on how mVP40 assembles to form the matrix layer following membrane binding. Here we have used hydrogen/deuterium exchange (HDX) mass spectrometry to determine the solvent accessibility of mVP40 residues in the absence and presence of phosphatidylserine and phosphatidylinositol 4,5-bisphosphate. HDX analysis demonstrates that two basic loops in the mVP40 C-terminal domain make important contributions to anionic membrane binding and also reveals a potential oligomerization interface in the C-terminal domain as well as a conserved oligomerization interface in the mVP40 N-terminal domain. Lipid binding assays confirm the role of the two basic patches elucidated with HD/X measurements, whereas molecular dynamics simulations and membrane insertion measurements complement these studies to demonstrate that mVP40 does not appreciably insert into the hydrocarbon region of anionic membranes in contrast to the matrix protein from Ebola virus. Taken together, we propose a model by which association of the mVP40 dimer with the anionic plasma membrane facilitates assembly of mVP40 oligomers. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Rotavirus architecture at subnanometer resolution.

PubMed

Li, Zongli; Baker, Matthew L; Jiang, Wen; Estes, Mary K; Prasad, B V Venkataram

2009-02-01

Rotavirus, a nonturreted member of the Reoviridae, is the causative agent of severe infantile diarrhea. The double-stranded RNA genome encodes six structural proteins that make up the triple-layer particle. X-ray crystallography has elucidated the structure of one of these capsid proteins, VP6, and two domains from VP4, the spike protein. Complementing this work, electron cryomicroscopy (cryoEM) has provided relatively low-resolution structures for the triple-layer capsid in several biochemical states. However, a complete, high-resolution structural model of rotavirus remains unresolved. Combining new structural analysis techniques with the subnanometer-resolution cryoEM structure of rotavirus, we now provide a more detailed structural model for the major capsid proteins and their interactions within the triple-layer particle. Through a series of intersubunit interactions, the spike protein (VP4) adopts a dimeric appearance above the capsid surface, while forming a trimeric base anchored inside one of the three types of aqueous channels between VP7 and VP6 capsid layers. While the trimeric base suggests the presence of three VP4 molecules in one spike, only hints of the third molecule are observed above the capsid surface. Beyond their interactions with VP4, the interactions between VP6 and VP7 subunits could also be readily identified. In the innermost T=1 layer composed of VP2, visualization of the secondary structure elements allowed us to identify the polypeptide fold for VP2 and examine the complex network of interactions between this layer and the T=13 VP6 layer. This integrated structural approach has resulted in a relatively high-resolution structural model for the complete, infectious structure of rotavirus, as well as revealing the subtle nuances required for maintaining interactions in such a large macromolecular assembly.

Structure of the Reston ebolavirus VP30 C-terminal domain.

PubMed

Clifton, Matthew C; Kirchdoerfer, Robert N; Atkins, Kateri; Abendroth, Jan; Raymond, Amy; Grice, Rena; Barnes, Steve; Moen, Spencer; Lorimer, Don; Edwards, Thomas E; Myler, Peter J; Saphire, Erica Ollmann

2014-04-01

The ebolaviruses can cause severe hemorrhagic fever. Essential to the ebolavirus life cycle is the protein VP30, which serves as a transcriptional cofactor. Here, the crystal structure of the C-terminal, NP-binding domain of VP30 from Reston ebolavirus is presented. Reston VP30 and Ebola VP30 both form homodimers, but the dimeric interfaces are rotated relative to each other, suggesting subtle inherent differences or flexibility in the dimeric interface.

Integrin-Using Rotaviruses Bind α2β1 Integrin α2 I Domain via VP4 DGE Sequence and Recognize αXβ2 and αVβ3 by Using VP7 during Cell Entry

PubMed Central

Graham, Kate L.; Halasz, Peter; Tan, Yan; Hewish, Marilyn J.; Takada, Yoshikazu; Mackow, Erich R.; Robinson, Martyn K.; Coulson, Barbara S.

2003-01-01

Integrins α2β1, αXβ2, and αVβ3 have been implicated in rotavirus cell attachment and entry. The virus spike protein VP4 contains the α2β1 ligand sequence DGE at amino acid positions 308 to 310, and the outer capsid protein VP7 contains the αXβ2 ligand sequence GPR. To determine the viral proteins and sequences involved and to define the roles of α2β1, αXβ2, and αVβ3, we analyzed the ability of rotaviruses and their reassortants to use these integrins for cell binding and infection and the effect of peptides DGEA and GPRP on these events. Many laboratory-adapted human, monkey, and bovine viruses used integrins, whereas all porcine viruses were integrin independent. The integrin-using rotavirus strains each interacted with all three integrins. Integrin usage related to VP4 serotype independently of sialic acid usage. Analysis of rotavirus reassortants and assays of virus binding and infectivity in integrin-transfected cells showed that VP4 bound α2β1, and VP7 interacted with αXβ2 and αVβ3 at a postbinding stage. DGEA inhibited rotavirus binding to α2β1 and infectivity, whereas GPRP binding to αXβ2 inhibited infectivity but not binding. The truncated VP5* subunit of VP4, expressed as a glutathione S-transferase fusion protein, bound the expressed α2 I domain. Alanine mutagenesis of D308 and G309 in VP5* eliminated VP5* binding to the α2 I domain. In a novel process, integrin-using viruses bind the α2 I domain of α2β1 via DGE in VP4 and interact with αXβ2 (via GPR) and αVβ3 by using VP7 to facilitate cell entry and infection. PMID:12941907

Crystal Structures of Major Envelope Proteins VP26 and VP28 from White Spot Syndrome Virus Shed Light on Their Evolutionary Relationship

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

Tang,X.; Wu, J.; Sivaraman, J.

2007-01-01

White spot syndrome virus (WSSV) is a virulent pathogen known to infect various crustaceans. It has bacilliform morphology with a tail-like appendage at one end. The envelope consists of four major proteins. Envelope structural proteins play a crucial role in viral infection and are believed to be the first molecules to interact with the host. Here, we report the localization and crystal structure of major envelope proteins VP26 and VP28 from WSSV at resolutions of 2.2 and 2.0 {angstrom}, respectively. These two proteins alone account for approximately 60% of the envelope, and their structures represent the first two structural envelopemore » proteins of WSSV. Structural comparisons among VP26, VP28, and other viral proteins reveal an evolutionary relationship between WSSV envelope proteins and structural proteins from other viruses. Both proteins adopt {beta}-barrel architecture with a protruding N-terminal region. We have investigated the localization of VP26 and VP28 using immunoelectron microscopy. This study suggests that VP26 and VP28 are located on the outer surface of the virus and are observed as a surface protrusion in the WSSV envelope, and this is the first convincing observation for VP26. Based on our studies combined with the literature, we speculate that the predicted N-terminal transmembrane region of VP26 and VP28 may anchor on the viral envelope membrane, making the core {beta}-barrel protrude outside the envelope, possibly to interact with the host receptor or to fuse with the host cell membrane for effective transfer of the viral infection. Furthermore, it is tempting to extend this host interaction mode to other structural viral proteins of similar structures. Our finding has the potential to extend further toward drug and vaccine development against WSSV.« less

Effects of Human Parvovirus B19 and Bocavirus VP1 Unique Region on Tight Junction of Human Airway Epithelial A549 Cells

PubMed Central

Chiu, Chun-Ching; Shi, Ya-Fang; Yang, Jiann-Jou; Hsiao, Yuan-Chao; Tzang, Bor-Show; Hsu, Tsai-Ching

2014-01-01

As is widely recognized, human parvovirus B19 (B19) and human bocavirus (HBoV) are important human pathogens. Obviously, both VP1 unique region (VP1u) of B19 and HBoV exhibit the secreted phospholipase A2 (sPLA2)-like enzymatic activity and are recognized to participate in the pathogenesis of lower respiratory tract illnesses. However, exactly how, both VP1u from B19 and HBoV affect tight junction has seldom been addressed. Therefore, this study investigates how B19-VP1u and HBoV-VP1u may affect the tight junction of the airway epithelial A549 cells by examining phospholipase A2 activity and transepithelial electrical resistance (TEER) as well as performing immunoblotting analyses. Experimental results indicate that TEER is more significantly decreased in A549 cells by treatment with TNF-α (10 ng), two dosages of B19-VP1u and BoV-VP1u (400 ng and 4000 ng) or bee venom PLA2 (10 ng) than that of the control. Accordingly, more significantly increased claudin-1 and decreased occludin are detected in A549 cells by treatment with TNF-α or both dosages of HBoV-VP1u than that of the control. Additionally, more significantly decreased Na+/K+ ATPase is observed in A549 cells by treatment with TNF-α, high dosage of B19-VP1u or both dosages of BoV-VP1u than that of the control. Above findings suggest that HBoV-VP1u rather than B19 VP1u likely plays more important roles in the disruption of tight junction in the airway tract. Meanwhile, this discrepancy appears not to be associated with the secreted phospholipase A2 (sPLA2)-like enzymatic activity. PMID:25268969

Cocaine Inhibition of Synaptic Transmission in the Ventral Pallidum Is Pathway-Specific and Mediated by Serotonin.

PubMed

Matsui, Aya; Alvarez, Veronica A

2018-06-26

The ventral pallidum (VP) is part of the basal ganglia circuitry and a target of both direct and indirect pathway projections from the nucleus accumbens. VP is important in cocaine reinforcement, and the firing of VP neurons is modulated in vivo during cocaine self-administration. This modulation of firing is thought to be indirect via cocaine actions on dopamine in the accumbens. Here, we show that cocaine directly inhibits synaptic transmission evoked by selective stimulation of indirect pathway projections to VP neurons. The inhibition is independent of dopamine receptor activation, absent in 5-HT1B knockout mice, and mimicked by a serotonin transporter (SERT) blocker. SERT-expressing neurons in dorsal raphe project to the VP. Optogenetic stimulation of these projections evokes serotonin transients and effectively inhibits GABAergic transmission to VP neurons. This study shows that cocaine increases endogenous serotonin in the VP to suppress synaptic transmission selectively from indirect pathway projections to VP neurons. Published by Elsevier Inc.

Recombinant VP1 protein of duck hepatitis virus 1 expressed in Pichia pastoris and its immunogenicity in ducks.

PubMed

Wang, C; Li, X K; Wu, T C; Wang, Y; Zhang, C J; Cheng, X C; Chen, P Y

2014-01-01

The VP1 gene of duck hepatitis virus type 1 (DHV-1) strain VJ09 was amplified by reverse transcription PCR from the liver of a duckling with clinical symptoms of viral hepatitis. The resulting VP1 cDNA was 720 bp in length and encoded a 240-amino-acid protein. In VP1 gene-based phylogenetic analysis, the VJ09 strain grouped with DHV-1 genotype C. The VP1 gene was inserted into the expression vector pPICZαA and expressed in Pichia pastoris. The expressed VP1 protein was purified and identified by western blot analysis. To evaluate the recombinant VP1's immunogenic potential in ducklings, the antibodies raised in the immunized ducklings were titrated by ELISA, and lymphocyte proliferation and virus neutralization assays were performed. The results show that the recombinant VP1 protein induced a significant immune response in ducklings and this could be a candidate for the development of a subunit vaccine against DHV-1 genotype C.

Recombinant VP1 protein as a potential marker for the diagnosis of acute hepatitis A virus infection.

PubMed

da Silva Junior, Haroldo Cid; da Silva, Edimilson Domingos; Lewis-Ximenez de Souza Rodrigues, Lia Laura; Medeiros, Marco Alberto

2017-07-01

Since hepatitis A virus (HAV) production is time-consuming and expensive, the use of recombinant proteins may represent an alternative source of antigens for diagnostic purposes. The present study aimed to express, purify and evaluate the potential of recombinant VP1 protein (rVP1) as a marker for the diagnosis of acute HAV infection. The rVP1 was expressed and purified successfully from Escherichia coli. The purified rVP1 was used to establish an in-house enzyme-linked immunosorbent assay (ELISA-rVP1) for detection of IgM antibodies in sera from HAV-positive patients. For a cut-off point of 0.351, the sensitivity and specificity of ELISA-rVP1 were 100.0% and 95.0%, respectively. These results indicate that rVP1 may be a useful antigen for detection of IgM antibodies against HAV. Copyright © 2017 Elsevier B.V. All rights reserved.

Targeting of rotavirus VP6 to DEC-205 induces protection against the infection in mice.

PubMed

Badillo-Godinez, O; Gutierrez-Xicotencatl, L; Plett-Torres, T; Pedroza-Saavedra, A; Gonzalez-Jaimes, A; Chihu-Amparan, L; Maldonado-Gama, M; Espino-Solis, G; Bonifaz, L C; Esquivel-Guadarrama, F

2015-08-20

Rotavirus (RV) is the primary etiologic agent of severe gastroenteritis in human infants. Although two attenuated RV-based vaccines have been licensed to be applied worldwide, they are not so effective in low-income countries, and the induced protection mechanisms have not been clearly established. Thus, it is important to develop new generation vaccines that induce long lasting heterotypic immunity. VP6 constitutes the middle layer protein of the RV virion. It is the most conserved protein and it is the target of protective T-cells; therefore, it is a potential candidate antigen for a new generation vaccine against the RV infection. We determined whether targeting the DEC-205 present in dendritic cells (DCs) with RV VP6 could induce protection at the intestinal level. VP6 was cross-linked to a monoclonal antibody (mAb) against murine DEC-205 (αDEC-205:VP6), and BALB/c mice were inoculated subcutaneously (s.c.) twice with the conjugated containing 1.5 μg of VP6 in the presence of polyinosinic-polycytidylic acid (Poly I:C) as adjuvant. As controls and following the same protocol, mice were immunized with ovalbumin (OVA) cross-linked to the mAb anti-DEC-205 (αDEC-205:OVA), VP6 cross-linked to a control isotype mAb (Isotype:VP6), 3 μg of VP6 alone, Poly I:C or PBS. Two weeks after the last inoculation, mice were orally challenged with a murine RV. Mice immunized with α-DEC-205:VP6 and VP6 alone presented similar levels of serum Abs to VP6 previous to the virus challenge. However, after the virus challenge, only α-DEC-205:VP6 induced up to a 45% IgA-independent protection. Memory T-helper (Th) cells from the spleen and the mesenteric lymph node (MLN) showed a Th1-type response upon antigen stimulation in vitro. These results show that when VP6 is administered parenterally targeting DEC-205, it can induce protection at the intestinal level at a very low dose, and this protection may be Th1-type cell dependent. Copyright © 2015 Elsevier Ltd. All rights reserved.

Structural and Functional Characterization of Reston Ebola Virus VP35 Interferon Inhibitory Domain

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

Leung, Daisy W.; Shabman, Reed S.; Farahbakhsh, Mina

2010-09-21

Ebolaviruses are causative agents of lethal hemorrhagic fever in humans and nonhuman primates. Among the filoviruses characterized thus far, Reston Ebola virus (REBOV) is the only Ebola virus that is nonpathogenic to humans despite the fact that REBOV can cause lethal disease in nonhuman primates. Previous studies also suggest that REBOV is less effective at inhibiting host innate immune responses than Zaire Ebola virus (ZEBOV) or Marburg virus. Virally encoded VP35 protein is critical for immune suppression, but an understanding of the relative contributions of VP35 proteins from REBOV and other filoviruses is currently lacking. In order to address thismore » question, we characterized the REBOV VP35 interferon inhibitory domain (IID) using structural, biochemical, and virological studies. These studies reveal differences in double-stranded RNA binding and interferon inhibition between the two species. These observed differences are likely due to increased stability and loss of flexibility in REBOV VP35 IID, as demonstrated by thermal shift stability assays. Consistent with this finding, the 1.71-{angstrom} crystal structure of REBOV VP35 IID reveals that it is highly similar to that of ZEBOV VP35 IID, with an overall backbone r.m.s.d. of 0.64 {angstrom}, but contains an additional helical element at the linker between the two subdomains of VP35 IID. Mutations near the linker, including swapping sequences between REBOV and ZEBOV, reveal that the linker sequence has limited tolerance for variability. Together with the previously solved ligand-free and double-stranded-RNA-bound forms of ZEBOV VP35 IID structures, our current studies on REBOV VP35 IID reinforce the importance of VP35 in immune suppression. Functional differences observed between REBOV and ZEBOV VP35 proteins may contribute to observed differences in pathogenicity, but these are unlikely to be the major determinant. However, the high level of similarity in structure and the low tolerance for sequence variability, coupled with the multiple critical roles played by Ebola virus VP35 proteins, highlight the viability of VP35 as a potential target for therapeutic development.« less

Structural and functional characterization of Reston Ebola virus VP35 interferon inhibitory domain.

PubMed

Leung, Daisy W; Shabman, Reed S; Farahbakhsh, Mina; Prins, Kathleen C; Borek, Dominika M; Wang, Tianjiao; Mühlberger, Elke; Basler, Christopher F; Amarasinghe, Gaya K

2010-06-11

Ebolaviruses are causative agents of lethal hemorrhagic fever in humans and nonhuman primates. Among the filoviruses characterized thus far, Reston Ebola virus (REBOV) is the only Ebola virus that is nonpathogenic to humans despite the fact that REBOV can cause lethal disease in nonhuman primates. Previous studies also suggest that REBOV is less effective at inhibiting host innate immune responses than Zaire Ebola virus (ZEBOV) or Marburg virus. Virally encoded VP35 protein is critical for immune suppression, but an understanding of the relative contributions of VP35 proteins from REBOV and other filoviruses is currently lacking. In order to address this question, we characterized the REBOV VP35 interferon inhibitory domain (IID) using structural, biochemical, and virological studies. These studies reveal differences in double-stranded RNA binding and interferon inhibition between the two species. These observed differences are likely due to increased stability and loss of flexibility in REBOV VP35 IID, as demonstrated by thermal shift stability assays. Consistent with this finding, the 1.71-A crystal structure of REBOV VP35 IID reveals that it is highly similar to that of ZEBOV VP35 IID, with an overall backbone r.m.s.d. of 0.64 A, but contains an additional helical element at the linker between the two subdomains of VP35 IID. Mutations near the linker, including swapping sequences between REBOV and ZEBOV, reveal that the linker sequence has limited tolerance for variability. Together with the previously solved ligand-free and double-stranded-RNA-bound forms of ZEBOV VP35 IID structures, our current studies on REBOV VP35 IID reinforce the importance of VP35 in immune suppression. Functional differences observed between REBOV and ZEBOV VP35 proteins may contribute to observed differences in pathogenicity, but these are unlikely to be the major determinant. However, the high level of similarity in structure and the low tolerance for sequence variability, coupled with the multiple critical roles played by Ebola virus VP35 proteins, highlight the viability of VP35 as a potential target for therapeutic development. Copyright 2010 Elsevier Ltd. All rights reserved.

Isolation and characterization of two VpYABBY genes from wild Chinese Vitis pseudoreticulata.

PubMed

Xiang, J; Liu, R Q; Li, T M; Han, L J; Zou, Y; Xu, T F; Wei, J Y; Wang, Y J; Xu, Y

2013-12-01

The establishment of abaxial-adaxial polarity is an important feature of the development of lateral organs in plants. Members of the YABBY gene family may be specific to seed-plant-specific transcriptional regulators that play critical roles in promoting abaxial cell fate in the model eudicot, Arabidopsis thaliana. However, recent study has shown that the roles of YABBY genes are not conserved in the development of angiosperms. The establishment of abaxial-adaxial polarity has not been studied in perennial fruit crops. Grapes are an important fruit crop in many regions of the world. Investigating YABBY genes in grapevines should help us to discover more about the key genetic and molecular pathways in grapevine development. To understand the characterization of YABBY genes in grapevines, two YABBY genes, VpYABBY1 (GenBank accession No. KC139089) and VpYABBY2 (GenBank accession No. KC139090), were isolated from the wild Chinese species Vitis pseudoreticulata. Both of these encode YABBY proteins. Sequence characterization and phylogenetic analyses show that VpYABBY1 is group classified into the FIL subfamily while VpYABBY2 is a member of the YAB2 subfamily of Arabidopsis thaliana. Subcellular localization analysis indicates that VpYABBY1 and VpYABBY2 proteins are localized in the nucleus. Tissue specific expressional analysis reveals that VpYABBY1 is expressed strongly in young leaves of grape but only weakly in the mature leaves. Meanwhile, VpYABBY2 is expressed in grape stems, flowers, tendrils, and leaves. Transgenic Arabidopsis plants ectopically expressing VpYABBY1 caused the partial abaxialization of the adaxial epidermises of leaves, behaving similarly to those over-expressing FIL or YAB3 with abaxialized lateral organs. By contrast, ectopic expression of VpYABBY2 in Arabidopsis did not cause any alteration in the adaxial-abaxial polarity. Sequence characterization and phylogenetic analysis revealed that VpYABBY1 and VpYABBY2 are group-classified into two different subfamilies. They have diverged functionally in the control of lateral organ development. VpYABBY1 may have a function in leaf development, while VpYABBY2 may play a specific role in carpel development and grape berry morphogenesis. It is further possible that during the evolution of different species, YABBY family members have preserved different expression regulatory systems and functions.

Interactions between the two surface proteins of rotavirus may alter the receptor-binding specificity of the virus.

PubMed Central

Méndez, E; Arias, C F; López, S

1996-01-01

The infection of target cells by most animal rotavirus strains requires the presence of sialic acids (SAs) on the cell surface. We recently isolated variants from simian rotavirus RRV whose infectivity is no longer dependent on SAs and showed that the mutant phenotype segregates with the gene coding for VP4, one of the two surface proteins of rotaviruses (the other one being VP7). The nucleotide sequence of the VP4 gene of four independently isolated variants showed three amino acid changes, at positions 37 (Leu to Pro), 187 (Lys to Arg), and 267 (Tyr to Cys), in all mutant VP4 proteins compared with RRV VP4. The characterization of revertant viruses from two independent mutants showed that the arginine residue at position 187 changed back to lysine, indicating that this amino acid is involved in the determination of the mutant phenotype. Surprisingly, sequence analysis of reassortant virus DS1XRRV, which depends on SAs to infect the cell, showed that its VP4 gene is identical to the VP4 gene of the variants. Since the only difference between DS1XRRV and the RRV variants is the parental origin of the VP7 gene (human rotavirus DS1 in the reassortant), these findings suggest that the receptor-binding specificity of rotaviruses, via VP4, may be influenced by the associated VP7 protein. PMID:8551583

Decreased LRIG1 in Human Ovarian Cancer Cell SKOV3 Upregulates MRP-1 and Contributes to the Chemoresistance of VP16.

PubMed

Yang, Hua; Yao, Jun; Yin, Jiangpin; Wei, Xuan

2016-05-01

The leucine-rich repeats and immunoglobulin-like domains (LRIG) are used as tumor suppressors in clinical applications. Although the LRIG has been identified to manipulate the cell proliferation via various oncogenic receptor tyrosine kinases in diverse cancers, its role in multidrug resistance needs to be further elucidated, especially in human ovarian cancer. We herein established that the etoposide (VP16)-resistant SKOV3 human ovarian cancer cell clones (SKOV3/VP16 cells) and mRNA expression of LRIG1 were significantly reduced by the treatment of VP16 in a concentration-dependent manner. Moreover, downregulated LRIG1 in SKOV3 could enhance the colony formation and resist the inhibition of proliferation by VP16, leading to the elevated expression of Bcl-2 and decreased apoptosis of SKOV3. Interestingly, our results uncovered that the multidrug resistance-associated protein 1 (MRP-1) was upregulated for the chemoresistance of VP16. To overcome the chemoresistance of SKOV3, SKOV3/VP16 was ectopically expressed of LRIG1. We found that the inhibition of VP16 on colony formation and proliferation was remarkably enhanced with increased apoptosis in SKOV3/VP16. Furthermore, the expression of MRP-1 and Bcl-2 was also inhibited, suggesting that the LRIG1could negatively control MRP-1 and the apoptosis to improve the sensitivity of VP16-related chemotherapy.

[Indirect ELISA for simultaneous detection of antibodies against duck hepatitis A type 1 and 3 viruses].

PubMed

Zhang, Yuyao; Ma, Xiuli; Huang, Bing; Li, Yufeng; Yu, Kexiang; Li, Jianliang; Liu, Cunxia; Han, Hongyu; Cui, Yanshun

2015-04-04

To simultaneously detect antibodies against Duck hepatitis A type 1 (DHAV-1) and type 3 (DHAV-3) viruses, we developed an indirect enzyme-linked immunosobent assay (ELISA) with bacterially expressed recombinant viral protein as antigen in Escherichia coli. We amplified the full-length VP3 gene of DHAV-1 and the full-length VP1 gene of DHAV-3 through reverse transcription-polymerase chain reaction (RT-PCR) and then cloned them into pET-32a expression vector, designated as pET-1VP3-3VP1. The fusion protein DHAV-1VP3-3VP1 expressed correctly and was subsequently used to develop an indirect ELISA assay. DHAV-1VP3-3VP1 fusion protein expressed in BL21 (DE3) cells following induction by Isopropyl-beta-D-1-thiogalactopyranoside (IPTG). The expressed protein was very antigenic and reactive to virus-specific antibodies in western blot assay. The optimal working concentration for coating antigen was 1.0 microg per well and the working concentration of serum samples was 1:200 dilution and the cut-off value that distinguished the positive from negative serum samples was OD650 > OR = 0.38. The ELISA method based on the prokaryotic expression of VP3 (DHAV-1) and VP1 proteins (DHAV-3) can be used effectively for the clinical detection antibodies against DHAV-1 and DHAV-3.

Live Attenuated Vaccine Based on Duck Enteritis Virus against Duck Hepatitis A Virus Types 1 and 3

PubMed Central

Zou, Zhong; Ma, Ji; Huang, Kun; Chen, Huanchun; Liu, Ziduo; Jin, Meilin

2016-01-01

As causative agents of duck viral hepatitis, duck hepatitis A virus type 1 (DHAV-1) and type 3 (DHAV-3) causes significant economic losses in the duck industry. However, a licensed commercial vaccine that simultaneously controls both pathogens is currently unavailable. Here, we generated duck enteritis virus recombinants (rC-KCE-2VP1) containing both VP1 from DHAV-1 (VP1/DHAV-1) and VP1 from DHAV-3 (VP1/DHAV-3) between UL27 and UL26. A self-cleaving 2A-element of FMDV was inserted between the two different types of VP1, allowing production of both proteins from a single open reading frame. Immunofluorescence and Western blot analysis results demonstrated that both VP1 proteins were robustly expressed in rC-KCE-2VP1-infected chicken embryo fibroblasts. Ducks that received a single dose of rC-KCE-2VP1 showed potent humoral and cellular immune responses and were completely protected against challenges of both pathogenic DHAV-1 and DHAV-3 strains. The protection was rapid, achieved as early as 3 days after vaccination. Moreover, viral replication was fully blocked in vaccinated ducks as early as 1 week post-vaccination. These results demonstrated, for the first time, that recombinant rC-KCE-2VP1 is potential fast-acting vaccine against DHAV-1 and DHAV-3. PMID:27777571

The VP40 protein of Marburg virus exhibits impaired budding and increased sensitivity to human tetherin following mouse adaptation.

PubMed

Feagins, Alicia R; Basler, Christopher F

2014-12-01

The Marburg virus VP40 protein is a viral matrix protein that spontaneously buds from cells. It also functions as an interferon (IFN) signaling antagonist by targeting Janus kinase 1 (JAK1). A previous study demonstrated that the VP40 protein of the Ravn strain of Marburg virus (Ravn virus [RAVV]) failed to block IFN signaling in mouse cells, whereas the mouse-adapted RAVV (maRAVV) VP40 acquired the ability to inhibit IFN responses in mouse cells. The increased IFN antagonist function of maRAVV VP40 mapped to residues 57 and 165, which were mutated during the mouse adaptation process. In the present study, we demonstrate that maRAVV VP40 lost the capacity to efficiently bud from human cell lines, despite the fact that both parental and maRAVV VP40s bud efficiently from mouse cell lines. The impaired budding in human cells corresponds with the appearance of protrusions on the surface of maRAVV VP40-expressing Huh7 cells and with an increased sensitivity of maRAVV VP40 to restriction by human tetherin but not mouse tetherin. However, transfer of the human tetherin cytoplasmic tail to mouse tetherin restored restriction of maRAVV VP40. Residues 57 and 165 were demonstrated to contribute to the failure of maRAVV VP40 to bud from human cells, and residue 57 was demonstrated to alter VP40 oligomerization, as assessed by coprecipitation assay, and to determine sensitivity to human tetherin. This suggests that RAVV VP40 acquired, during adaptation to mice, changes in its oligomerization potential that enhanced IFN antagonist function. However, this new capacity impaired RAVV VP40 budding from human cells. Filoviruses, which include Marburg viruses and Ebola viruses, are zoonotic pathogens that cause severe disease in humans and nonhuman primates but do not cause similar disease in wild-type laboratory strains of mice unless first adapted to these animals. Although mouse adaptation has been used as a method to develop small animal models of pathogenesis, the molecular determinants associated with filovirus mouse adaptation are poorly understood. Our study demonstrates how genetic changes that accrued during mouse adaptation of the Ravn strain of Marburg virus have impacted the budding function of the viral VP40 matrix protein. Strikingly, we find impairment of mouse-adapted VP40 budding function in human but not mouse cell lines, and we correlate the impairment with an increased sensitivity of VP40 to restriction by human but not mouse tetherin and with changes in VP40 oligomerization. These data suggest that there are functional costs associated with filovirus adaptation to new hosts and implicate tetherin as a filovirus host restriction factor. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Abscisic acid and stress signals induce Viviparous1 expression in seed and vegetative tissues of maize.

PubMed

Cao, Xueyuan; Costa, Liliana M; Biderre-Petit, Corinne; Kbhaya, Bouchab; Dey, Nrisingha; Perez, Pascual; McCarty, Donald R; Gutierrez-Marcos, Jose F; Becraft, Philip W

2007-02-01

Viviparous1 (Vp1) encodes a B3 domain-containing transcription factor that is a key regulator of seed maturation in maize (Zea mays). However, the mechanisms of Vp1 regulation are not well understood. To examine physiological factors that may regulate Vp1 expression, transcript levels were monitored in maturing embryos placed in culture under different conditions. Expression of Vp1 decreased after culture in hormone-free medium, but was induced by salinity or osmotic stress. Application of exogenous abscisic acid (ABA) also induced transcript levels within 1 h in a dose-dependent manner. The Vp1 promoter fused to beta-glucuronidase or green fluorescent protein reproduced the endogenous Vp1 expression patterns in transgenic maize plants and also revealed previously unknown expression domains of Vp1. The Vp1 promoter is active in the embryo and aleurone cells of developing seeds and, upon drought stress, was also found in phloem cells of vegetative tissues, including cobs, leaves, and stems. Sequence analysis of the Vp1 promoter identified a potential ABA-responsive complex, consisting of an ACGT-containing ABA response element (ABRE) and a coupling element 1-like motif. Electrophoretic mobility shift assay confirmed that the ABRE and putative coupling element 1 components specifically bound proteins in embryo nuclear protein extracts. Treatment of embryos in hormone-free Murashige and Skoog medium blocked the ABRE-protein interaction, whereas exogenous ABA or mannitol treatment restored this interaction. Our data support a model for a VP1-dependent positive feedback mechanism regulating Vp1 expression during seed maturation.

The hepatitis E virus open reading frame 3 product interacts with microtubules and interferes with their dynamics.

PubMed

Kannan, Harilakshmi; Fan, Sumin; Patel, Deendayal; Bossis, Ioannis; Zhang, Yan-Jin

2009-07-01

Hepatitis E virus (HEV) is the causative agent of hepatitis E, a major form of viral hepatitis in developing countries. The open reading frame 3 (ORF3) of HEV encodes a phosphoprotein with a molecular mass of approximately 13 kDa (hereinafter called vp13). vp13 is essential for establishing HEV infections in animals, yet its exact functions are still obscure. Our current study found evidence showing interaction between vp13 and microtubules. Live-cell confocal fluorescence microscopy revealed both filamentous and punctate distribution patterns of vp13 in cells transfected with recombinant ORF3 reporter plasmids. The filamentous pattern of vp13 was altered by a microtubule-destabilizing drug. The vp13 expression led to elevation of acetylated alpha-tubulin, indicating increased microtubule stability. Its association with microtubules was further supported by its presence in microtubule-containing pellets in microtubule isolation assays. Exposure of these pellets to a high-salt buffer caused release of the vp13 to the supernatant, suggesting an electrostatic interaction. Inclusion of ATP and GTP in the lysis buffer during microtubule isolation also disrupted the interaction, indicating its sensitivity to the nucleotides. Further assays showed that motor proteins are needed for the vp13 association with the microtubules because disruption of dynein function abolished the vp13 filamentous pattern. Analysis of ORF3 deletion constructs found that both of the N-terminal hydrophobic domains of vp13 are needed for the interaction. Thus, our findings suggest that the vp13 interaction with microtubules might be needed for establishment of an HEV infection.

Human parvovirus B19 VP1u Protein as inflammatory mediators induces liver injury in naïve mice.

PubMed

Hsu, Tsai-Ching; Chiu, Chun-Ching; Chang, Shun-Chih; Chan, Hsu-Chin; Shi, Ya-Fang; Chen, Tzy-Yen; Tzang, Bor-Show

2016-01-01

Human parvovirus B19 (B19V) is a human pathogen known to be associated with many non-erythroid diseases, including hepatitis. Although B19V VP1-unique region (B19-VP1u) has crucial roles in the pathogenesis of B19V infection, the influence of B19-VP1u proteins on hepatic injury is still obscure. This study investigated the effect and possible inflammatory signaling of B19-VP1u in livers from BALB/c mice that were subcutaneously inoculated with VP1u-expressing COS-7 cells. The in vivo effects of B19-VP1u were analyzed by using live animal imaging system (IVIS), Haematoxylin-Eosin staining, gel zymography, and immunoblotting after inoculation. Markedly hepatocyte disarray and lymphocyte infiltration, enhanced matrix metalloproteinase (MMP)-9 activity and increased phosphorylation of p38, ERK, IKK-α, IκB and NF-κB (p-p65) proteins were observed in livers from BALB/c mice receiving COS-7 cells expressing B19-VP1u as well as the significantly increased CRP, IL-1β and IL-6. Notably, IFN-γ and phosphorylated STAT1, but not STAT3, were also significantly increased in the livers of BALB/c mice that were subcutaneously inoculated with VP1u-expressing COS-7 cells. These findings revealed the effects of B19-VP1u on liver injury and suggested that B19-VP1u may have a role as mediators of inflammation in B19V infection.

Addition of six-His-tagged peptide to the C terminus of adeno-associated virus VP3 does not affect viral tropism or production.

PubMed

Zhang, Huang-Ge; Xie, Jinfu; Dmitriev, Igor; Kashentseva, Elena; Curiel, David T; Hsu, Hui-Chen; Mountz, John D

2002-12-01

Production of large quantities of recombinant adeno-associated virus (AAV) is difficult and not cost-effective. To overcome this problem, we have explored the feasibility of creating a recombinant AAV encoding a 6xHis tag on the VP3 capsid protein. We generated a plasmid vector containing a six-His (6xHis)-tagged AAV VP3. A second plasmid vector was generated that contained the full-length AAV capsid capable of producing VP1 and VP2, but not VP3 due to a mutation at position 2809 that encodes the start codon for VP3. These plasmids, necessary for production of AAV, were transfected into 293 cells to generate a 6xHis-tagged VP3mutant recombinant AAV. The 6xHis-tagged VP3 did not affect the formation of AAV virus, and the physical properties of the 6xHis-modified AAV were equivalent to those of wild-type particles. The 6xHis-tagged AAV did not affect the production titer of recombinant AAV and could be used to purify the recombinant AAV using an Ni-nitrilotriacetic acid column. Addition of the 6xHis tag did not alter the viral tropism compared to wild-type AAV. These observations demonstrate the feasibility of producing high-titer AAV containing a 6xHis-tagged AAV VP3 capsid protein and to utilize the 6xHis-tagged VP3 capsid to achieve high-affinity purification of this recombinant AAV.

A novel parallel pipeline structure of VP9 decoder

NASA Astrophysics Data System (ADS)

Qin, Huabiao; Chen, Wu; Yi, Sijun; Tan, Yunfei; Yi, Huan

2018-04-01

To improve the efficiency of VP9 decoder, a novel parallel pipeline structure of VP9 decoder is presented in this paper. According to the decoding workflow, VP9 decoder can be divided into sub-modules which include entropy decoding, inverse quantization, inverse transform, intra prediction, inter prediction, deblocking and pixel adaptive compensation. By analyzing the computing time of each module, hotspot modules are located and the causes of low efficiency of VP9 decoder can be found. Then, a novel pipeline decoder structure is designed by using mixed parallel decoding methods of data division and function division. The experimental results show that this structure can greatly improve the decoding efficiency of VP9.

Preliminary molecular detection of the somatic embryogenesis receptor-like kinase (VpSERK) and knotted-like homeobox (VpKNOX1) genes during in vitro morphogenesis of Vanilla planifolia Jacks.

PubMed

Ramírez-Mosqueda, Marco A; Iglesias-Andreu, Lourdes G; Sáenz, Luis; Córdova, Iván

2018-02-01

This work aimed to evaluate the embryogenic competence of different tissues from different stages (friable callus, bud-regenerating callus, and whole buds) of Vanilla planifolia , through the molecular detection of the somatic embryogenesis receptor-like kinase ( VpSERK ) and knotted-like homeobox ( VpKNOX1 ) genes. RNA was extracted with Trizol ® , cDNA was obtained, and the studied transcripts were amplified. Using non-specific primers, VpSERK and VpSTM gene expression was detected in the three stages evaluated. This study might contribute to providing an explanation for the recalcitrance of this Vanilla species to somatic embryogenesis.

MRI of the sacroiliac joints in spondyloarthritis: the added value of intra-articular signal changes for a 'positive MRI'.

PubMed

Laloo, Frederiek; Herregods, N; Jaremko, J L; Verstraete, K; Jans, L

2018-05-01

To determine if intra-articular signal changes at the sacroiliac joint space on MRI have added diagnostic value for spondyloarthritis, when compared to bone marrow edema (BME). A retrospective study was performed on the MRIs of sacroiliac joints of 363 patients, aged 16-45 years, clinically suspected of sacroiliitis. BME of the sacroiliac joints was correlated to intra-articular sacroiliac joint MR signal changes: high T1 signal, fluid signal, ankylosis and vacuum phenomenon (VP). These MRI findings were correlated with final clinical diagnosis. Sensitivity (SN), specificity (SP), likelihood ratios (LR), predictive values and post-test probabilities were calculated. BME had SN of 68.9%, SP of 74.0% and LR+ of 2.6 for diagnosis of spondyloarthritis. BME in absence of intra-articular signal changes had a lower SN and LR+ for spondyloarthritis (SN = 20.5%, LR+ 1.4). Concomitant BME and high T1 signal (SP = 97.2%, LR + = 10.5), BME and fluid signal (SP = 98.6%, LR + = 10.3) or BME and ankylosis (SP = 100%) had higher SP and LR+ for spondyloarthritis. Concomitant BME and VP had low LR+ for spondyloarthritis (SP = 91%, LR + =0.9). When BME was absent, intra-articular signal changes were less prevalent, but remained highly specific for spondyloarthritis. Our results suggest that both periarticular and intra-articular MR signal of the sacroiliac joint should be examined to determine whether an MRI is 'positive' or 'not positive' for sacroiliitis associated with spondyloarthritis.

Simulation of rarefied low pressure RF plasma flow around the sample

NASA Astrophysics Data System (ADS)

Zheltukhin, V. S.; Shemakhin, A. Yu

2017-01-01

The paper describes a mathematical model of the flow of radio frequency plasma at low pressure. The hybrid mathematical model includes the Boltzmann equation for the neutral component of the RF plasma, the continuity and the thermal equations for the charged component. Initial and boundary conditions for the corresponding equations are described. The electron temperature in the calculations is 1-4 eV, atoms temperature in the plasma clot is (3-4) • 103 K, in the plasma jet is (3.2-10) • 102 K, the degree of ionization is 10-7-10-5, electron density is 1015-1019 m-3. For calculations plasma parameters is developed soft package on C++ program language, that uses the OpenFOAM library package. Simulations for the vacuum chamber in the presence of a sample and the free jet flow were carried out.

Refrigerated poultry breast fillets packed in modified atmosphere and irradiated: bacteriological evaluation, shelf life and sensory acceptance

PubMed Central

Mantilla, Samira Pirola Santos; Santos, Érica Barbosa; de Freitas, Mônica Queiroz; de Carvalho Vital, Helio; Mano, Sérgio Borges; Franc, Robson Maia

2012-01-01

In the present study the effects on shelf life and sensory acceptance of gamma-irradiated refrigerated poultry breast fillets subjected to modified atmosphere packaging (80% CO2/20% N2 or vacuum) were investigated. After irradiation with 2 kGy, sensory acceptance tests and monitoring of bacterial growth were performed in order to determine the sanitary quality of the samples. It has been found that irradiation, used in combination with modified atmosphere packaging, can double the shelf life of refrigerated poultry breast fillets by reducing the populations of aerobic mesophilic and psychrotrophic bacteria, enterobacteria, coliforms, Listeria spp. and Aeromonas spp., without significantly modifying its color or its overall appearance, the lactic acid bacteria being the most resistant to exposure to radiation and carbon dioxide. PMID:24031967

A method for encapsulating high voltage power transformers

NASA Astrophysics Data System (ADS)

Sanchez, Robert O.

Voltage breakdowns become a major concern in reducing the size of high-voltage power converter transformers. Even the smallest of voids can provide a path for corona discharge which can cause a dielectric breakdown leading to a transformer failure. A method of encapsulating small high voltage transformers has been developed. The method virtually eliminates voids in the impregnation material, provides an exceptional dielectric between windings and provides a mechanically rugged package. The encapsulation material is a carboxyl terminated butadiene nitril (CTBN) modified mica filled epoxy. The method requires heat/vacuum to impregnate the coil and heat/pressure to cure the encapsulant. The transformer package utilizes a diallyl phthalate (DAP) contact assembly in which a coated core/coil assembly is mounted and soldered. This assembly is then loaded into an RTV mold and the encapsulation process begins.

WOLF: a computer code package for the calculation of ion beam trajectories

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

Vogel, D.L.

1985-10-01

The WOLF code solves POISSON'S equation within a user-defined problem boundary of arbitrary shape. The code is compatible with ANSI FORTRAN and uses a two-dimensional Cartesian coordinate geometry represented on a triangular lattice. The vacuum electric fields and equipotential lines are calculated for the input problem. The use may then introduce a series of emitters from which particles of different charge-to-mass ratios and initial energies can originate. These non-relativistic particles will then be traced by WOLF through the user-defined region. Effects of ion and electron space charge are included in the calculation. A subprogram PISA forms part of this codemore » and enables optimization of various aspects of the problem. The WOLF package also allows detailed graphics analysis of the computed results to be performed.« less

Structural insights into the multifunctional protein VP3 of birnaviruses.

PubMed

Casañas, Arnau; Navarro, Aitor; Ferrer-Orta, Cristina; González, Dolores; Rodríguez, José F; Verdaguer, Núria

2008-01-01

Infectious bursal disease virus (IBDV), a member of the Birnaviridae family, is the causative agent of one of the most harmful poultry diseases. The IBDV genome encodes five mature proteins; of these, the multifunctional protein VP3 plays an essential role in virus morphogenesis. This protein, which interacts with the structural protein VP2, with the double-stranded RNA genome, and with the virus-encoded, RNA-dependent RNA polymerase, VP1, is involved not only in the formation of the viral capsid, but also in the recruitment of VP1 into the capsid and in the encapsidation of the viral genome. Here, we report the X-ray structure of the central region of VP3, residues 92-220, consisting of two alpha-helical domains connected by a long and flexible hinge that are organized as a dimer. Unexpectedly, the overall fold of the second VP3 domain shows significant structural similarities with different transcription regulation factors.

Comparison of two eukaryotic systems for the expression of VP6 protein of rotavirus specie A: transient gene expression in HEK293-T cells and insect cell-baculovirus system.

PubMed

da Silva Junior, Haroldo Cid; da Silva E Mouta Junior, Sérgio; de Mendonça, Marcos César Lima; de Souza Pereira, Mirian Claudia; da Rocha Nogueira, Alanderson; de Azevedo, Maria Luiza Borges; Leite, José Paulo Gagliardi; de Moraes, Márcia Terezinha Baroni

2012-09-01

The VP6 protein of rotavirus A (RVA) is a target antigen used for diagnostic assays and also for the development of new RVA vaccines. We have compared the expression of VP6 protein in human embryonic kidney (HEK293-T) cells with results obtained using a well-established insect cell-baculovirus system. The recombinant VP6 (rVP6) expressed in HEK293-T cells did not present degradation and also retained the ability to form trimers. In the insect cell-baculovirus system, rVP6 was expressed at higher levels and with protein degradation as well as partial loss of ability to form trimers was observed. Therefore, HEK293-T cells represent a less laborious alternative system than insect cells for expression of rVP6 from human RVA.

An overview of new video coding tools under consideration for VP10: the successor to VP9

NASA Astrophysics Data System (ADS)

Mukherjee, Debargha; Su, Hui; Bankoski, James; Converse, Alex; Han, Jingning; Liu, Zoe; Xu, Yaowu

2015-09-01

Google started an opensource project, entitled the WebM Project, in 2010 to develop royaltyfree video codecs for the web. The present generation codec developed in the WebM project called VP9 was finalized in mid2013 and is currently being served extensively by YouTube, resulting in billions of views per day. Even though adoption of VP9 outside Google is still in its infancy, the WebM project has already embarked on an ambitious project to develop a next edition codec VP10 that achieves at least a generational bitrate reduction over the current generation codec VP9. Although the project is still in early stages, a set of new experimental coding tools have already been added to baseline VP9 to achieve modest coding gains over a large enough test set. This paper provides a technical overview of these coding tools.

Impact of Nisin-Activated Packaging on Microbiota of Beef Burgers during Storage.

PubMed

Ferrocino, Ilario; Greppi, Anna; La Storia, Antonietta; Rantsiou, Kalliopi; Ercolini, Danilo; Cocolin, Luca

2016-01-15

Beef burgers were stored at 4°C in a vacuum in nisin-activated antimicrobial packaging. Microbial ecology analyses were performed on samples collected between days 0 and 21 of storage to discover the population diversity. Two batches were analyzed using RNA-based denaturing gradient gel electrophoresis (DGGE) and pyrosequencing. The active packaging retarded the growth of the total viable bacteria and lactic acid bacteria. Culture-independent analysis by pyrosequencing of RNA extracted directly from meat showed that Photobacterium phosphoreum, Lactococcus piscium, Lactobacillus sakei, and Leuconostoc carnosum were the major operational taxonomic units (OTUs) shared between control and treated samples. Beta diversity analysis of the 16S rRNA sequence data and RNA-DGGE showed a clear separation between two batches based on the microbiota. Control samples from batch B showed a significant high abundance of some taxa sensitive to nisin, such as Kocuria rhizophila, Staphylococcus xylosus, Leuconostoc carnosum, and Carnobacterium divergens, compared to control samples from batch A. However, only from batch B was it possible to find a significant difference between controls and treated samples during storage due to the active packaging. Predicted metagenomes confirmed differences between the two batches and indicated that the use of nisin-based antimicrobial packaging can determine a reduction in the abundance of specific metabolic pathways related to spoilage. The present study aimed to assess the viable bacterial communities in beef burgers stored in nisin-based antimicrobial packaging, and it highlights the efficacy of this strategy to prolong beef burger shelf life. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Generation of a parvovirus B19 vaccine candidate.

PubMed

Chandramouli, Sumana; Medina-Selby, Angelica; Coit, Doris; Schaefer, Mary; Spencer, Terika; Brito, Luis A; Zhang, Pu; Otten, Gillis; Mandl, Christian W; Mason, Peter W; Dormitzer, Philip R; Settembre, Ethan C

2013-08-20

Parvovirus B19 is the causative agent of fifth disease in children, aplastic crisis in those with blood dyscrasias, and hydrops fetalis. Previous parvovirus B19 virus-like-particle (VLP) vaccine candidates were produced by co-infection of insect cells with two baculoviruses, one expressing wild-type VP1 and the other expressing VP2. In humans, the VLPs were immunogenic but reactogenic. We have developed new VLP-based parvovirus B19 vaccine candidates, produced by co-expressing VP2 and either wild-type VP1 or phospholipase-negative VP1 in a regulated ratio from a single plasmid in Saccharomyces cerevisiae. These VLPs are expressed efficiently, are very homogeneous, and can be highly purified. Although VP2 alone can form VLPs, in mouse immunizations, VP1 and the adjuvant MF59 are required to elicit a neutralizing response. Wild-type VLPs and those with phospholipase-negative VP1 are equivalently potent. The purity, homogeneity, yeast origin, and lack of phospholipase activity of these VLPs address potential causes of previously observed reactogenicity. Copyright © 2013 Elsevier Ltd. All rights reserved.

Mutual antagonism between the Ebola virus VP35 protein and the RIG-I activator PACT determines infection outcome.

PubMed

Luthra, Priya; Ramanan, Parameshwaran; Mire, Chad E; Weisend, Carla; Tsuda, Yoshimi; Yen, Benjamin; Liu, Gai; Leung, Daisy W; Geisbert, Thomas W; Ebihara, Hideki; Amarasinghe, Gaya K; Basler, Christopher F

2013-07-17

The cytoplasmic pattern recognition receptor RIG-I is activated by viral RNA and induces type I IFN responses to control viral replication. The cellular dsRNA binding protein PACT can also activate RIG-I. To counteract innate antiviral responses, some viruses, including Ebola virus (EBOV), encode proteins that antagonize RIG-I signaling. Here, we show that EBOV VP35 inhibits PACT-induced RIG-I ATPase activity in a dose-dependent manner. The interaction of PACT with RIG-I is disrupted by wild-type VP35, but not by VP35 mutants that are unable to bind PACT. In addition, PACT-VP35 interaction impairs the association between VP35 and the viral polymerase, thereby diminishing viral RNA synthesis and modulating EBOV replication. PACT-deficient cells are defective in IFN induction and are insensitive to VP35 function. These data support a model in which the VP35-PACT interaction is mutually antagonistic and plays a fundamental role in determining the outcome of EBOV infection. Copyright © 2013 Elsevier Inc. All rights reserved.

Simulation and flavor compound analysis of dealcoholized beer via one-step vacuum distillation.

PubMed

Andrés-Iglesias, Cristina; García-Serna, Juan; Montero, Olimpio; Blanco, Carlos A

2015-10-01

The coupled operation of vacuum distillation process to produce alcohol free beer at laboratory scale and Aspen HYSYS simulation software was studied to define the chemical changes during the dealcoholization process in the aroma profiles of 2 different lager beers. At the lab-scale process, 2 different parameters were chosen to dealcoholize beer samples, 102mbar at 50°C and 200mbar at 67°C. Samples taken at different steps of the process were analyzed by HS-SPME-GC-MS focusing on the concentration of 7 flavor compounds, 5 alcohols and 2 esters. For simulation process, the EoS parameters of the Wilson-2 property package were adjusted to the experimental data and one more pressure was tested (60mbar). Simulation methods represent a viable alternative to predict results of the volatile compound composition of a final dealcoholized beer. Copyright © 2015 Elsevier Ltd. All rights reserved.

Protoflight photovoltaic power module system-level tests in the space power facility

NASA Technical Reports Server (NTRS)

Rivera, Juan C.; Kirch, Luke A.

1989-01-01

Work Package Four, which includes the NASA-Lewis and Rocketdyne, has selected an approach for the Space Station Freedom Photovoltaic (PV) Power Module flight certification that combines system level qualification and acceptance testing in the thermal vacuum environment: The protoflight vehicle approach. This approach maximizes ground test verification to assure system level performance and to minimize risk of on-orbit failures. The preliminary plans for system level thermal vacuum environmental testing of the protoflight PV Power Module in the NASA-Lewis Space Power Facility (SPF), are addressed. Details of the facility modifications to refurbish SPF, after 13 years of downtime, are briefly discussed. The results of an evaluation of the effectiveness of system level environmental testing in screening out incipient part and workmanship defects and unique failure modes are discussed. Preliminary test objectives, test hardware configurations, test support equipment, and operations are presented.

Test Package Plummets in the Zero Gravity Research Facility

NASA Image and Video Library

1966-09-21

National Aeronautics and Space Administration (NASA) Lewis Research Center. Lewis researchers had been studying the behavior of liquid in microgravity for several years using ballistic rocket flights, aircraft flying series of parabolas, and in the 2.2-Second Drop Tower. It was easier to control experiments and repeat tests based on almost instantaneous test results in the Zero Gravity Research Facility than missiles or aircraft. It also more than doubled the microgravity time of the original drop tower. The experiments were enclosed in a large experiment package that was suspended inside the chamber. A vacuum was introduced to the chamber before the package was released. The test equipment allowed researchers to film and take measurements of the experiment as it was falling. The 2500‐pound package was slowed by special Styrofoam‐like pellets in a decelerator cart. An experiment, traveling 176 feet per second, was stopped in about 15 feet of deceleration material. The facility’s designers struggled to determine the correct type of deceleration pellets to use. For several years Lewis engineers tested various samples from manufacturers. The final selection was not made until the facility’s completion in May 1966, just before the facility made its public debut at the 1966 Inspection of the Center.

Retention of heavy metal ions on comb-type hydrogels based on acrylic acid and 4-vinylpyridine, synthesized by gamma radiation

NASA Astrophysics Data System (ADS)

González-Gómez, Roberto; Ortega, Alejandra; Lazo, Luz M.; Burillo, Guillermina

2014-09-01

Two novel comb-type hydrogels based on pH-sensitive monomers (acrylic acid (AAc) and 4-vinylpyridine (4VP) were synthesized by gamma radiation. The systems were as follows: a) comb-type hydrogels of an AAc network followed by grafting of 4VP ((net-PAAc)-g-4VP) and b) comb-type hydrogels of an AAc network grafted onto polypropylene (PP) followed by grafting of 4VP (net-(PP-g-AAc)-g-4VP). The equilibrium isotherms and kinetics were evaluated for copper and zinc ions in aqueous solutions. The Zn(II) retention obtained was 480 mg g-1 and 1086 mg g-1 for (net-PAAc)-g-4VP and net-(PP-g-AAc)-g-4VP, respectively. At concentrations as low as ppm, retention efficiencies of approximately 90% were achieved for Cu(II) on (net-PAAc)-g-4VP and for Zn(II) on net-(PP-g-AAc)-g-4VP. Desorption of the hydrogels was also studied, and the results indicated that they can be used repeatedly in aqueous solutions. For both systems, the adsorption of Cu(II) and Zn(II) obeyed the Freundlich model, indicating heterogeneous sorption, and the retention process occurred by chemisorption. The sorption process follows a pseudo-second-order model.

High-yield production of the VP1 structural protein epitope from serotype O foot-and-mouth disease virus in Escherichia coli.

PubMed

Jung, Joon-Goo; Lee, Yong Jae; Velmurugan, Natarajan; Ko, Young-Joon; Lee, Hyang-Sim; Jeong, Ki Jun

2013-07-01

For effective control of foot-and-mouth disease (FMD), the development of rapid diagnostic systems and vaccines are required against its etiological agent, FMD virus (FMDV). To accomplish this, efficient large-scale expression of the FMDV VP1 protein, with high solubility, needs to be optimized. We attempted to produce high levels of a serotype O FMDV VP1 epitope in Escherichia coli. We identified the subtype-independent serotype O FMDV VP1 epitope sequence and used it to construct a glutathione S-transferase (GST) fusion protein. For efficient production of the FMDV VP1 epitope fused to GST (VP1e-GST), four E. coli strains and three temperatures were examined. The conditions yielding the greatest level of VP1e-GST with highest solubility were achieved with E. coli BL21(DE3) at 25 °C. For high-level production, fed-batch cultures were conducted in 5-l bioreactors. When cells were induced at a high density and complex feeding solutions were supplied, approximately 11 g of VP1e-GST was obtained from a 2.9-l culture. Following purification, the VP1 epitope was used to immunize rabbits, and we confirmed that it induced an immune response.

The ebola virus interferon antagonist VP24 directly binds STAT1 and has a novel, pyramidal fold.

PubMed

Zhang, Adrianna P P; Bornholdt, Zachary A; Liu, Tong; Abelson, Dafna M; Lee, David E; Li, Sheng; Woods, Virgil L; Saphire, Erica Ollmann

2012-02-01

Ebolaviruses cause hemorrhagic fever with up to 90% lethality and in fatal cases, are characterized by early suppression of the host innate immune system. One of the proteins likely responsible for this effect is VP24. VP24 is known to antagonize interferon signaling by binding host karyopherin α proteins, thereby preventing them from transporting the tyrosine-phosphorylated transcription factor STAT1 to the nucleus. Here, we report that VP24 binds STAT1 directly, suggesting that VP24 can suppress at least two distinct branches of the interferon pathway. Here, we also report the first crystal structures of VP24, derived from different species of ebolavirus that are pathogenic (Sudan) and nonpathogenic to humans (Reston). These structures reveal that VP24 has a novel, pyramidal fold. A site on a particular face of the pyramid exhibits reduced solvent exchange when in complex with STAT1. This site is above two highly conserved pockets in VP24 that contain key residues previously implicated in virulence. These crystal structures and accompanying biochemical analysis map differences between pathogenic and nonpathogenic viruses, offer templates for drug design, and provide the three-dimensional framework necessary for biological dissection of the many functions of VP24 in the virus life cycle.

Insights into the homo-oligomerization properties of N-terminal coiled-coil domain of Ebola virus VP35 protein.

PubMed

Ramaswamy, Venkata Krishnan; Di Palma, Francesco; Vargiu, Attilio V; Corona, Angela; Piano, Dario; Ruggerone, Paolo; Zinzula, Luca; Tramontano, Enzo

2018-03-02

The multifunctional Ebola virus (EBOV) VP35 protein is a key determinant of virulence. VP35 is essential for EBOV replication, is a component of the viral RNA polymerase and participates in nucleocapsid formation. Furthermore, VP35 contributes to EBOV evasion of the host innate immune response by suppressing RNA silencing and blocking RIG-I like receptors' pathways that lead to type I interferon (IFN) production. VP35 homo-oligomerization has been reported to be critical for its replicative function and to increase its IFN-antagonism properties. Moreover, homo-oligomerization is mediated by a predicted coiled-coil (CC) domain located within its N-terminal region. Here we report the homo-oligomerization profile of full-length recombinant EBOV VP35 (rVP35) assessed by size-exclusion chromatography and native polyacrylamide gel electrophoresis. Based on our biochemical results and in agreement with previous experimental observations, we have built an in silico 3D model of the so-far structurally unsolved EBOV VP35 CC domain and performed self-assembly homo-oligomerization simulations by means of molecular dynamics. Our model advances the understanding of how VP35 may associate in different homo-oligomeric species, a crucial process for EBOV replication and pathogenicity. Copyright © 2018 Elsevier B.V. All rights reserved.

Forecasting the Human Pathogen Vibrio Parahaemolyticus in Shellfish Tissue within Long Island Sound

NASA Astrophysics Data System (ADS)

Whitney, M. M.; DeRosia-Banick, K.

2016-02-01

Vibrio parahaemolyticus (Vp) is a marine bacterium that occurs naturally in brackish and saltwater environments and may be found in higher concentrations in the warmest months. Vp is a growing threat to producing safe seafood. Consumption of shellfish with high Vp levels can result in gastrointestinal human illnesses. Management response to Vp-related illness outbreaks includes closure of shellfish growing areas. Water quality observations, Vp measurements, and model forecasts are key components to effective management of shellfish growing areas. There is a clear need for observations within the growing area themselves. These areas are offshore of coastal stations and typically inshore of the observing system moorings. New field observations in Long Island Sound (LIS) shellfish growing areas are described and their agreement with high-resolution satellite sea surface temperature data is discussed. A new dataset of Vp concentrations in shellfish tissue is used to determine the LIS-specific Vp vs. temperature relationship following methods in the FDA pre-harvest Vp risk model. This information is combined with output from a high-resolution hydrodynamic model of LIS to make daily forecasts of Vp levels. The influence of river inflows, the role of heat waves, and predictions for future warmer climates are discussed. The key elements of this observational-modeling approach to pathogen forecasting are extendable to other coastal systems.

Amino Acid Residue at Position 79 of Marburg Virus VP40 Confers Interferon Antagonism in Mouse Cells.

PubMed

Feagins, Alicia R; Basler, Christopher F

2015-10-01

Marburg viruses (MARVs) cause highly lethal infections in humans and nonhuman primates. Mice are not generally susceptible to MARV infection; however, if the strain is first adapted to mice through serial passaging, it becomes able to cause disease in this animal. A previous study correlated changes accrued during mouse adaptation in the VP40 gene of a MARV strain known as Ravn virus (RAVV) with an increased capacity to inhibit interferon (IFN) signaling in mouse cell lines. The MARV strain Ci67, which belongs to a different phylogenetic clade than RAVV, has also been adapted to mice and in the process the Ci67 VP40 acquired a different collection of genetic changes than did RAVV VP40. Here, we demonstrate that the mouse-adapted Ci67 VP40 more potently antagonizes IFN-α/β-induced STAT1 and STAT2 tyrosine phosphorylation, gene expression, and antiviral activity in both mouse and human cell lines, compared with the parental Ci67 VP40. Ci67 VP40 is also demonstrated to target the activation of kinase Jak1. A single change at VP40 residue 79 was found to be sufficient for the increased VP40 IFN antagonism. These data argue that VP40 IFN-antagonist activity plays a key role in MARV pathogenesis in mice. © The Author 2015. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Shrimp arginine kinase being a binding protein of WSSV envelope protein VP31

NASA Astrophysics Data System (ADS)

Ma, Cuiyan; Gao, Qiang; Liang, Yan; Li, Chen; Liu, Chao; Huang, Jie

2016-11-01

Viral entry into the host is the earliest stage of infection in the viral life cycle in which attachment proteins play a key role. VP31 (WSV340/WSSV396), an envelope protein of white spot syndrome virus (WSSV), contains an Arg-Gly-Asp (RGD) peptide domain known as a cellular attachment site. At present, the process of VP31 interacting with shrimp host cells has not been explored. Therefore, the VP31 gene was cloned into pET30a (+), expressed in Escherichia coli strain BL21 and purified with immobilized metal ion affinity chromatography. Four gill cellular proteins of shrimp ( Fenneropenaeus chinensis) were pulled down by an affinity column coupled with recombinant VP31 (rVP31), and the amino acid sequences were identified with MALDI-TOF/TOF mass spectrometry. Hemocyanin, beta-actin, arginine kinase (AK), and an unknown protein were suggested as the putative VP31 receptor proteins. SDS-PAGE showed that AK is the predominant binding protein of VP31. An i n vitro binding activity experiment indicated that recombinant AK's (rAK) binding activity with rVP31 is comparable to that with the same amount of WSSV. These results suggested that AK, as a member of the phosphagen kinase family, plays a role in WSSV infection. This is the first evidence showing that AK is a binding protein of VP31. Further studies on this topic will elucidate WSSV infection mechanism in the future.

Expression, Purification, Crystallization of Two Major Envelope Proteins from White Spot Syndrome Virus

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

Tang,X.; Hew, C.

2007-01-01

White spot syndrome virus (WSSV) is a major virulent pathogen known to infect penaeid shrimp and other crustaceans. VP26 and VP28, two major envelope proteins from WSSV, have been identified and overexpressed in Escherichia coli. In order to facilitate purification and crystallization, predicted N-terminal transmembrane regions of approximately 35 amino acids have been truncated from both VP26 and VP28. Truncated VP26 and VP28 and their corresponding SeMet-labelled proteins were purified and the SeMet proteins were crystallized by the hanging-drop vapor-diffusion method. Crystals of SeMet-labelled VP26 were obtained using a reservoir consisting of 0.1 M citric acid pH 3.5, 3.0 Mmore » sodium chloride and 1%(w/v) polyethylene glycol 3350, whereas SeMet VP28 was crystallized using a reservoir solution consisting of 25% polyethylene glycol 8000, 0.2 M calcium acetate, 0.1 M Na HEPES pH 7.5 and 1.5%(w/v) 1,2,3-heptanetriol. Crystals of SeMet-labelled VP26 diffract to 2.2 {angstrom} resolution and belong to space group R32, with unit-cell parameters a = b = 73.92, c = 199.31 {angstrom}. SeMet-labelled VP28 crystallizes in space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 105.33, b = 106.71, c = 200.37 {angstrom}, and diffracts to 2.0 {angstrom} resolution.« less

Pros and cons of VP1-specific maternal IgG for the protection of Enterovirus 71 infection.

PubMed

Kim, Young-In; Song, Jae-Hyoung; Kwon, Bo-Eun; Kim, Ha-Neul; Seo, Min-Duk; Park, KwiSung; Lee, SangWon; Yeo, Sang-Gu; Kweon, Mi-Na; Ko, Hyun-Jeong; Chang, Sun-Young

2015-11-27

Enterovirus 71 (EV71) causes hand, foot, and mouth diseases and can result in severe neurological disorders when it infects the central nervous system. Thus, there is a need for the development of effective vaccines against EV71 infection. Here we report that viral capsid protein 1 (VP1), one of the main capsid proteins of EV71, efficiently elicited VP1-specific immunoglobulin G (IgG) in the serum of mice immunized with recombinant VP1. The VP1-specific IgG produced in female mice was efficiently transferred to their offspring, conferring protection against EV71 infection immediately after birth. VP1-specific antibody can neutralize EV71 infection and protect host cells. VP1-specific maternal IgG in offspring was maintained for over 6 months. However, the pre-existence of VP1-specific maternal IgG interfered with the production of VP1-specific IgG antibody secreting cells by active immunization in offspring. Therefore, although our results showed the potential for VP1-specific maternal IgG protection against EV71 in neonatal mice, other strategies must be developed to overcome the hindrance of maternal IgG in active immunization. In this study, we developed an effective and feasible animal model to evaluate the protective efficacy of humoral immunity against EV71 infection using a maternal immunity concept. Copyright © 2015 Elsevier Ltd. All rights reserved.

VP3 is crucial for the stability of Nora virus virions.

PubMed

Sadanandan, Sajna Anand; Ekström, Jens-Ola; Jonna, Venkateswara Rao; Hofer, Anders; Hultmark, Dan

2016-09-02

Nora virus is an enteric virus that causes persistent, non-pathological infection in Drosophila melanogaster. It replicates in the fly gut and is transmitted via the fecal-oral route. Nora virus has a single-stranded positive-sense RNA genome, which is translated in four open reading frames. Reading frame three encodes the VP3 protein, the structure and function of which we have investigated in this work. We have shown that VP3 is a trimer that has an α-helical secondary structure, with a functionally important coiled-coil domain. In order to identify the role of VP3 in the Nora virus life cycle, we constructed VP3-mutants using the cDNA clone of the virus. Our results show that VP3 does not have a role in the actual assembly of the virus particles, but virions that lack VP3 or harbor VP3 with a disrupted coiled coil domain are incapable of transmission via the fecal-oral route. Removing the region downstream of the putative coiled coil appears to have an effect on the fitness of the virus but does not hamper its replication or transmission. We also found that the VP3 protein and particularly the coiled coil domain are crucial for the stability of Nora virus virions when exposed to heat or proteases. Hence, we propose that VP3 is imperative to Nora virus virions as it confers stability to the viral capsid. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

Conditional poliovirus mutants made by random deletion mutagenesis of infectious cDNA.

PubMed Central

Kirkegaard, K; Nelsen, B

1990-01-01

Small deletions were introduced into DNA plasmids bearing cDNA copies of Mahoney type 1 poliovirus RNA. The procedure used was similar to that of P. Hearing and T. Shenk (J. Mol. Biol. 167:809-822, 1983), with modifications designed to introduce only one lesion randomly into each DNA molecule. Methods to map small deletions in either large DNA or RNA molecules were employed. Two poliovirus mutants, VP1-101 and VP1-102, were selected from mutagenized populations on the basis of their host range phenotype, showing a large reduction in the relative numbers of plaques on CV1 and HeLa cells compared with wild-type virus. The deletions borne by the mutant genomes were mapped to the region encoding the amino terminus of VP1. That these lesions were responsible for the mutant phenotypes was substantiated by reintroduction of the sequenced lesions into a wild-type poliovirus cDNA by deoxyoligonucleotide-directed mutagenesis. The deletion of nucleotides encoding amino acids 8 and 9 of VP1 was responsible for the VP1-101 phenotype; the VP1-102 defect was caused by the deletion of the sequences encoding the first four amino acids of VP1. The peptide sequence at the VP1-VP3 proteolytic cleavage site was altered from glutamine-glycine to glutamine-methionine in VP1-102; this apparently did not alter the proteolytic cleavage pattern. The biochemical defects resulting from these mutations are discussed in the accompanying report. Images PMID:2152811

Fatty acids, sterols, and antioxidant activity in minimally processed avocados during refrigerated storage.

PubMed

Plaza, Lucía; Sánchez-Moreno, Concepción; de Pascual-Teresa, Sonia; de Ancos, Begoña; Cano, M Pilar

2009-04-22

Avocado ( Persea americana Mill.) is a good source of bioactive compounds such as monounsaturated fatty acids and sterols. The impact of minimal processing on its health-promoting attributes was investigated. Avocados cut into slices or halves were packaged in plastic bags under nitrogen, air, or vacuum and stored at 8 degrees C for 13 days. The stabilities of fatty acids and sterols as well as the effect on antioxidant activity were evaluated. The main fatty acid identified and quantified in avocado was oleic acid (about 57% of total content), whereas beta-sitosterol was found to be the major sterol (about 89% of total content). In general, after refrigerated storage, a significant decrease in fatty acid content was observed. Vacuum/halves and air/slices were the samples that maintained better this content. With regard to phytosterols, there were no significant changes during storage. Antioxidant activity showed a slight positive correlation against stearic acid content. At the end of refrigerated storage, a significant increase in antiradical efficiency (AE) was found for vacuum samples. AE values were quite similar among treatments. Hence, minimal processing can be a useful tool to preserve health-related properties of avocado fruit.

Three-dimensional seismic velocity structure of Mauna Loa and Kilauea volcanoes in Hawaii from local seismic tomography

USGS Publications Warehouse

Lin, Guoqing; Shearer, Peter M.; Matoza, Robin S.; Okubo, Paul G.; Amelung, Falk

2016-01-01

We present a new three-dimensional seismic velocity model of the crustal and upper mantle structure for Mauna Loa and Kilauea volcanoes in Hawaii. Our model is derived from the first-arrival times of the compressional and shear waves from about 53,000 events on and near the Island of Hawaii between 1992 and 2009 recorded by the Hawaiian Volcano Observatory stations. The Vp model generally agrees with previous studies, showing high-velocity anomalies near the calderas and rift zones and low-velocity anomalies in the fault systems. The most significant difference from previous models is in Vp/Vs structure. The high-Vp and high-Vp/Vs anomalies below Mauna Loa caldera are interpreted as mafic magmatic cumulates. The observed low-Vp and high-Vp/Vs bodies in the Kaoiki seismic zone between 5 and 15 km depth are attributed to the underlying volcaniclastic sediments. The high-Vp and moderate- to low-Vp/Vs anomalies beneath Kilauea caldera can be explained by a combination of different mafic compositions, likely to be olivine-rich gabbro and dunite. The systematically low-Vp and low-Vp/Vs bodies in the southeast flank of Kilauea may be caused by the presence of volatiles. Another difference between this study and previous ones is the improved Vp model resolution in deeper layers, owing to the inclusion of events with large epicentral distances. The new velocity model is used to relocate the seismicity of Mauna Loa and Kilauea for improved absolute locations and ultimately to develop a high-precision earthquake catalog using waveform cross-correlation data.

The use of additive and subtractive approaches to examine the nuclear localization sequence of the polyomavirus major capsid protein VP1

NASA Technical Reports Server (NTRS)

Chang, D.; Haynes, J. I. 2nd; Brady, J. N.; Consigli, R. A.; Spooner, B. S. (Principal Investigator)

1992-01-01

A nuclear localization signal (NLS) has been identified in the N-terminal (Ala1-Pro-Lys-Arg-Lys-Ser-Gly-Val-Ser-Lys-Cys11) amino acid sequence of the polyomavirus major capsid protein VP1. The importance of this amino acid sequence for nuclear transport of VP1 protein was demonstrated by a genetic "subtractive" study using the constructs pSG5VP1 (full-length VP1) and pSG5 delta 5'VP1 (truncated VP1, lacking amino acids Ala1-Cys11). These constructs were used to transfect COS-7 cells, and expression and intracellular localization of the VP1 protein was visualized by indirect immunofluorescence. These studies revealed that the full-length VP1 was expressed and localized in the nucleus, while the truncated VP1 protein was localized in the cytoplasm and not transported to the nucleus. These findings were substantiated by an "additive" approach using FITC-labeled conjugates of synthetic peptides homologous to the NLS of VP1 cross-linked to bovine serum albumin or immunoglobulin G. Both conjugates localized in the nucleus after microinjection into the cytoplasm of 3T6 cells. The importance of individual amino acids found in the basic sequence (Lys3-Arg-Lys5) of the NLS was also investigated. This was accomplished by synthesizing three additional peptides in which lysine-3 was substituted with threonine, arginine-4 was substituted with threonine, or lysine-5 was substituted with threonine. It was found that lysine-3 was crucial for nuclear transport, since substitution of this amino acid with threonine prevented nuclear localization of the microinjected, FITC-labeled conjugate.

Low-tube voltage 100 kVp MDCT in screening of cocaine body packing: image quality and radiation dose compared to 120 kVp MDCT.

PubMed

Aissa, Joel; Rubbert, Christian; Boos, Johannes; Schleich, Christoph; Thomas, Christoph; Kröpil, Patric; Antoch, Gerald; Miese, Falk

2015-10-01

The aim of this study was to evaluate the impact of a reduced tube potential (100 kVp) for non-enhanced abdominal low-dose CT on radiation dose and image quality (IQ) in the detection of body packing. This retrospective study was approved by the local research ethics committee of our clinic. From March 2012 to July 2014, 99 subjects were referred to our institute with suspected body packing. 50 CT scans were performed using a 120 kVp protocol (group A), and 49 CTs were performed using a low-dose protocol with a tube voltage of 100 kVp (group B). Subjective and objective IQ were assessed. DLP and CTDIvol were analyzed. All examinations were of diagnostic IQ. Objective IQ was not significantly different between the 120 kVp and 100 kVp protocol. Mean density of solid and liquid body packets was 210 ± 60.2 HU at 120 kVp and 250.6 ± 29.7 HU at 100 kVp. Radiation dose was significantly lower in group B as compared to group A (p < 0.05). In group A, body packs were detected in 16 (32%) of the 50 patients. In group B, packets were observed in 15 (31%) of 49 patients. Laboratory analysis detected cocaine in all smuggled body packs. Low-tube voltage 100 kVp MDCT with automated tube current modulation in screening of illegal drugs leads to a diagnostic IQ and significant dose reduction compared to 120 kVp low-tube voltage protocols. Despite lower radiation dose, liquid and solid cocaine containers retain high attenuation and are easily detected.

Membrane association and localization dynamics of the Ebola virus matrix protein VP40.

PubMed

Gc, Jeevan B; Gerstman, Bernard S; Chapagain, Prem P

2017-10-01

The Ebola virus matrix protein VP40 is a major structural protein that provides the scaffolding for new Ebola virus particles. For this, VP40 is first trafficked to the lower leaflet of the plasma membrane (PM) in its dimeric form. Once associated with the PM, the VP40 dimers undergo structural rearrangements and oligomerize into hexamers and filaments that make up the virus matrix. Therefore, association of the VP40 dimers and their stabilization at the PM is a crucial step in the Ebola life-cycle. To understand the molecular details of the VP40 dimer-PM interactions, we investigated the dimer association with the inner leaflet of the PM using detailed all-atom molecular dynamics (MD) simulations. The formation of the dimer-PM complex is facilitated by the interactions of the VP40 lysine residues and the anionic lipids POPS, POPI, and PIP 2 in the PM. In contrast, the dimer fails to associate with a membrane without POPS, POPI, or PIP 2 lipids. We explored the mechanisms of the association and identified important residues and lipids involved in localization and stabilization of VP40 dimers at the PM. MD simulations elucidate the role of a C-terminal α-helix alignment parallel to the lipid bilayer surface as well as the creation of membrane defects that allow partial insertion of the hydrophobic residue V276 into the membrane to further stabilize the VP40 dimer-PM complex. Understanding the mechanisms of the VP40 dimer-PM association that facilitate oligomerization can be important for potentially targeting the VP40 for small molecules that can interfere with the virus life-cycle. Copyright © 2017 Elsevier B.V. All rights reserved.

VP40 of the Ebola Virus as a Target for EboV Therapy: Comprehensive Conformational and Inhibitor Binding Landscape from Accelerated Molecular Dynamics.

PubMed

Balmith, Marissa; Soliman, Mahmoud E S

2017-03-01

The first account of the dynamic features of the loop region of VP40 of the Ebola virus was studied using accelerated molecular dynamics simulations and reported herein. Among the proteins of the Ebola virus, the matrix protein (VP40) plays a significant role in the virus lifecycle thereby making it a promising therapeutic target. Of interest is the newly elucidated N-terminal domain loop region of VP40 comprising residues K127, T129, and N130 which when mutated to alanine have demonstrated an unrecognized role for N-terminal domain-plasma membrane interaction for efficient VP40-plasma membrane localization, oligomerization, matrix assembly, and egress. The molecular understanding of the conformational features of VP40 in complex with a known inhibitor still remains elusive. Using accelerated molecular dynamics approaches, we conducted a comparative study on VP40 apo and bound systems to understand the conformational features of VP40 at the molecular level and to determine the effect of inhibitor binding with the aid of a number of post-dynamic analytical tools. Significant features were seen in the presence of an inhibitor as per molecular mechanics/generalized born surface area binding free energy calculations. Results revealed that inhibitor binding to VP40 reduces the flexibility and mobility of the protein as supported by root mean square fluctuation and root mean square deviation calculations. The study revealed a characteristic "twisting" motion and coiling of the loop region of VP40 accompanied by conformational changes in the dimer interface upon inhibitor binding. We believe that results presented in this study will ultimately provide useful insight into the binding landscape of VP40 which could assist researchers in the discovery of potent Ebola virus inhibitors for anti-Ebola therapies.

Chaperone-Mediated Autophagy Protein BAG3 Negatively Regulates Ebola and Marburg VP40-Mediated Egress

PubMed Central

Liang, Jingjing; Sagum, Cari A.; Bedford, Mark T.; Sudol, Marius; Han, Ziying

2017-01-01

Ebola (EBOV) and Marburg (MARV) viruses are members of the Filoviridae family which cause outbreaks of hemorrhagic fever. The filovirus VP40 matrix protein is essential for virus assembly and budding, and its PPxY L-domain motif interacts with WW-domains of specific host proteins, such as Nedd4 and ITCH, to facilitate the late stage of virus-cell separation. To identify additional WW-domain-bearing host proteins that interact with VP40, we used an EBOV PPxY-containing peptide to screen an array of 115 mammalian WW-domain-bearing proteins. Using this unbiased approach, we identified BCL2 Associated Athanogene 3 (BAG3), a member of the BAG family of molecular chaperone proteins, as a specific VP40 PPxY interactor. Here, we demonstrate that the WW-domain of BAG3 interacts with the PPxY motif of both EBOV and MARV VP40 and, unexpectedly, inhibits budding of both eVP40 and mVP40 virus-like particles (VLPs), as well as infectious VSV-EBOV recombinants. BAG3 is a stress induced protein that regulates cellular protein homeostasis and cell survival through chaperone-mediated autophagy (CMA). Interestingly, our results show that BAG3 alters the intracellular localization of VP40 by sequestering VP40 away from the plasma membrane. As BAG3 is the first WW-domain interactor identified that negatively regulates budding of VP40 VLPs and infectious virus, we propose that the chaperone-mediated autophagy function of BAG3 represents a specific host defense strategy to counteract the function of VP40 in promoting efficient egress and spread of virus particles. PMID:28076420

Chaperone-Mediated Autophagy Protein BAG3 Negatively Regulates Ebola and Marburg VP40-Mediated Egress.

PubMed

Liang, Jingjing; Sagum, Cari A; Bedford, Mark T; Sidhu, Sachdev S; Sudol, Marius; Han, Ziying; Harty, Ronald N

2017-01-01

Ebola (EBOV) and Marburg (MARV) viruses are members of the Filoviridae family which cause outbreaks of hemorrhagic fever. The filovirus VP40 matrix protein is essential for virus assembly and budding, and its PPxY L-domain motif interacts with WW-domains of specific host proteins, such as Nedd4 and ITCH, to facilitate the late stage of virus-cell separation. To identify additional WW-domain-bearing host proteins that interact with VP40, we used an EBOV PPxY-containing peptide to screen an array of 115 mammalian WW-domain-bearing proteins. Using this unbiased approach, we identified BCL2 Associated Athanogene 3 (BAG3), a member of the BAG family of molecular chaperone proteins, as a specific VP40 PPxY interactor. Here, we demonstrate that the WW-domain of BAG3 interacts with the PPxY motif of both EBOV and MARV VP40 and, unexpectedly, inhibits budding of both eVP40 and mVP40 virus-like particles (VLPs), as well as infectious VSV-EBOV recombinants. BAG3 is a stress induced protein that regulates cellular protein homeostasis and cell survival through chaperone-mediated autophagy (CMA). Interestingly, our results show that BAG3 alters the intracellular localization of VP40 by sequestering VP40 away from the plasma membrane. As BAG3 is the first WW-domain interactor identified that negatively regulates budding of VP40 VLPs and infectious virus, we propose that the chaperone-mediated autophagy function of BAG3 represents a specific host defense strategy to counteract the function of VP40 in promoting efficient egress and spread of virus particles.

Low-cost compact thermal imaging sensors for body temperature measurement

NASA Astrophysics Data System (ADS)

Han, Myung-Soo; Han, Seok Man; Kim, Hyo Jin; Shin, Jae Chul; Ahn, Mi Sook; Kim, Hyung Won; Han, Yong Hee

2013-06-01

This paper presents a 32x32 microbolometer thermal imaging sensor for human body temperature measurement. Waferlevel vacuum packaging technology allows us to get a low cost and compact imaging sensor chip. The microbolometer uses V-W-O film as sensing material and ROIC has been designed 0.35-um CMOS process in UMC. A thermal image of a human face and a hand using f/1 lens convinces that it has a potential of human body temperature for commercial use.

A Novel Vacuum Packaging Design Process for Microelectromechanical System (MEMS) Quad-Mass Gyroscopes

DTIC Science & Technology

2016-09-01

16 2.2.1 Bake Out 18 2.2.2 Ramp-Up 19 2.2.3 Lid Seal 20 2.2.4 Cool/Purge 22 2.3 Residual Gas Analyzer Experiment 23 3. Results 28 4...Fig. 24 RGA experiment setup .........................................................................24 Fig. 25 RGA output for a 1-h bake out at T...225 °C ......................................25 Fig. 26 RGA output of a 2-h sample for an 8-h bake out .................................26 Fig. 27

Crystal Structure of the Marburg Virus VP35 Oligomerization Domain.

PubMed

Bruhn, Jessica F; Kirchdoerfer, Robert N; Urata, Sarah M; Li, Sheng; Tickle, Ian J; Bricogne, Gérard; Saphire, Erica Ollmann

2017-01-15

Marburg virus (MARV) is a highly pathogenic filovirus that is classified in a genus distinct from that of Ebola virus (EBOV) (genera Marburgvirus and Ebolavirus, respectively). Both viruses produce a multifunctional protein termed VP35, which acts as a polymerase cofactor, a viral protein chaperone, and an antagonist of the innate immune response. VP35 contains a central oligomerization domain with a predicted coiled-coil motif. This domain has been shown to be essential for RNA polymerase function. Here we present crystal structures of the MARV VP35 oligomerization domain. These structures and accompanying biophysical characterization suggest that MARV VP35 is a trimer. In contrast, EBOV VP35 is likely a tetramer in solution. Differences in the oligomeric state of this protein may explain mechanistic differences in replication and immune evasion observed for MARV and EBOV. Marburg virus can cause severe disease, with up to 90% human lethality. Its genome is concise, only producing seven proteins. One of the proteins, VP35, is essential for replication of the viral genome and for evasion of host immune responses. VP35 oligomerizes (self-assembles) in order to function, yet the structure by which it assembles has not been visualized. Here we present two crystal structures of this oligomerization domain. In both structures, three copies of VP35 twist about each other to form a coiled coil. This trimeric assembly is in contrast to tetrameric predictions for VP35 of Ebola virus and to known structures of homologous proteins in the measles, mumps, and Nipah viruses. Distinct oligomeric states of the Marburg and Ebola virus VP35 proteins may explain differences between them in polymerase function and immune evasion. These findings may provide a more accurate understanding of the mechanisms governing VP35's functions and inform the design of therapeutics. Copyright © 2017 American Society for Microbiology.

Investigation of the Lipid Binding Properties of the Marburg Virus Matrix Protein VP40.

PubMed

Wijesinghe, Kaveesha J; Stahelin, Robert V

2015-12-30

Marburg virus (MARV), which belongs to the virus family Filoviridae, causes hemorrhagic fever in humans and nonhuman primates that is often fatal. MARV is a lipid-enveloped virus that during the replication process extracts its lipid coat from the plasma membrane of the host cell it infects. MARV carries seven genes, one of which encodes its matrix protein VP40 (mVP40), which regulates the assembly and budding of the virions. Currently, little information is available on mVP40 lipid binding properties. Here, we have investigated the in vitro and cellular mechanisms by which mVP40 associates with lipid membranes. mVP40 associates with anionic membranes in a nonspecific manner that is dependent upon the anionic charge density of the membrane. These results are consistent with recent structural determination of mVP40, which elucidated an mVP40 dimer with a flat and extensive cationic lipid binding interface. Marburg virus (MARV) is a lipid-enveloped filamentous virus from the family Filoviridae. MARV was discovered in 1967, and yet little is known about how its seven genes are used to assemble and form a new viral particle in the host cell it infects. The MARV matrix protein VP40 (mVP40) underlies the inner leaflet of the virus and regulates budding from the host cell plasma membrane. In vitro and cellular assays in this study investigated the mechanism by which mVP40 associates with lipids. The results demonstrate that mVP40 interactions with lipid vesicles or the inner leaflet of the plasma membrane are electrostatic but nonspecific in nature and are dependent on the anionic charge density of the membrane surface. Small molecules that can disrupt lipid trafficking or reduce the anionic charge of the plasma membrane interface may be useful in inhibiting assembly and budding of MARV. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Acutely elevated vasopressin increases circulating concentrations of cortisol and aldosterone in fasting northern elephant seal (Mirounga angustirostris) pups

NASA Technical Reports Server (NTRS)

Ortiz, Rudy M.; Wade, Charles E.; Ortiz, C. Leo; Talamantes, Frank

2003-01-01

The physiological actions of vasopressin (VP) in marine mammals are not well defined. To help elucidate its hormonal and renal effects in this group of mammals, northern elephant seal (Mirounga angustirostris) pups (N=7; 99+/-4 kg) were first infused with 0.9% saline (control; 220 ml), followed 24 h later with VP (as a 20 ng kg(-1) bolus, then 2 ng kg(-1) min(-1) for approximately 35 min in 225+/-16 ml saline). During both control and VP periods, blood samples were collected prior to infusion, and 15, 30, 60, 120 min and 24 h after infusion to examine the hormonal responses of the pups to VP. Renal responses were quantified from 24 h urine samples obtained prior to infusion (control) and 24 h post-infusion. Compared to the control period, infusion of VP increased plasma concentrations of cortisol over a 120 min period and aldosterone over 30 min, while plasma renin activity (PRA) was decreased for a 120 min period. The plasma urea:creatinine ratio was elevated following infusion of VP. Urine output and osmotic clearance were increased by 69+/-18% (mean +/- S.E.M.) and 36+/-10%, respectively, but free water clearance and glomerular filtration rate were not significantly altered 24 h post-infusion of VP. Solute (osmolality, Na(+), K(+) and Cl(-)) excretion and fractional excretion of electrolytes were also increased when compared to control values. The increase in cortisol concentration suggests that VP may possess corticotropin releasing hormone-like activity in elephant seals. If osmotic diuresis and natriuresis are typical consequences of elevated [VP] in fasting pups, then not increasing VP normally during the fast may serve as a protective mechanism to avoid the potential loss of Na(+) induced by elevated [VP]. Therefore, under natural fasting conditions, pups may be highly sensitive to small changes in [VP], resulting in the maintenance of water and electrolyte balance.

An interference account of the missing-VP effect

PubMed Central

Häussler, Jana; Bader, Markus

2015-01-01

Sentences with doubly center-embedded relative clauses in which a verb phrase (VP) is missing are sometimes perceived as grammatical, thus giving rise to an illusion of grammaticality. In this paper, we provide a new account of why missing-VP sentences, which are both complex and ungrammatical, lead to an illusion of grammaticality, the so-called missing-VP effect. We propose that the missing-VP effect in particular, and processing difficulties with multiply center-embedded clauses more generally, are best understood as resulting from interference during cue-based retrieval. When processing a sentence with double center-embedding, a retrieval error due to interference can cause the verb of an embedded clause to be erroneously attached into a higher clause. This can lead to an illusion of grammaticality in the case of missing-VP sentences and to processing complexity in the case of complete sentences with double center-embedding. Evidence for an interference account of the missing-VP effect comes from experiments that have investigated the missing-VP effect in German using a speeded grammaticality judgments procedure. We review this evidence and then present two new experiments that show that the missing-VP effect can be found in German also with less restricting procedures. One experiment was a questionnaire study which required grammaticality judgments from participants without imposing any time constraints. The second experiment used a self-paced reading procedure and did not require any judgments. Both experiments confirm the prior findings of missing-VP effects in German and also show that the missing-VP effect is subject to a primacy effect as known from the memory literature. Based on this evidence, we argue that an account of missing-VP effects in terms of interference during cue-based retrieval is superior to accounts in terms of limited memory resources or in terms of experience with embedded structures. PMID:26136698

Production and characterization of egg yolk antibody (IgY) against recombinant VP8-S2 antigen.

PubMed

Nasiri, K; Nassiri, M R; Tahmoorespur, M; Haghparast, A; Zibaee, S

2016-01-01

Bovine Rotavirus and Bovine Coronavirus are the most important causes of diarrhea in newborn calves and in some other species such as pigs and sheep. VP8 subunit of rotavirus is the major determinant of the viral infectivity and neutralization. Spike glycoprotein of coronavirus is responsible for induction of neutralizing antibody response. Studies showed that immunoglobulin of egg yolk (IgY) from immunized hens has been identified to be a convenient source for specific antibodies for using in immunotherapy and immunodiagnostic to limit the infections. In this study, chimeric VP8-S2 gene was designed using by computational techniques. The chimeric VP8-S2 gene was cloned and sub-cloned into pGH and pET32a (+) vectors. Then, recombinant pET32a-VP8-S2 vector was transferred into E. coli BL21 CodonPlus (DE3). The expressed protein was purified by Ni-NTA chromatography column. Hens were immunized with the purified VP8-S2 protein three times. IgY was purified from egg yolks using polyethylene glycol precipitation method. Activity and specificity of anti-VP8-S2 IgY were detected by dot-blotting, Western-blotting and indirect ELISA. We obtained anti-VP8-S2 IgY by immunizing hens with the recombinant VP8-S2 protein. The anti-VP8-S2 IgY was showed to bind specifically to the chimeric VP8-S2 protein by dot-blotting, Western-blotting analyses and indirect ELISA. The result of this study indicated that such construction can be useful to investigate as candidates for development of detection methods for simultaneous diagnosis of both infections. Specific IgY against the recombinant VP8-S2 could be recommended as a candidate for passive immunization against bovine rotavirus and bovine coronavirus.

Crystal Structure of the Marburg Virus VP35 Oligomerization Domain

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

Bruhn, Jessica F.; Kirchdoerfer, Robert N.; Urata, Sarah M.

ABSTRACT Marburg virus (MARV) is a highly pathogenic filovirus that is classified in a genus distinct from that of Ebola virus (EBOV) (generaMarburgvirusandEbolavirus, respectively). Both viruses produce a multifunctional protein termed VP35, which acts as a polymerase cofactor, a viral protein chaperone, and an antagonist of the innate immune response. VP35 contains a central oligomerization domain with a predicted coiled-coil motif. This domain has been shown to be essential for RNA polymerase function. Here we present crystal structures of the MARV VP35 oligomerization domain. These structures and accompanying biophysical characterization suggest that MARV VP35 is a trimer. In contrast, EBOVmore » VP35 is likely a tetramer in solution. Differences in the oligomeric state of this protein may explain mechanistic differences in replication and immune evasion observed for MARV and EBOV. IMPORTANCEMarburg virus can cause severe disease, with up to 90% human lethality. Its genome is concise, only producing seven proteins. One of the proteins, VP35, is essential for replication of the viral genome and for evasion of host immune responses. VP35 oligomerizes (self-assembles) in order to function, yet the structure by which it assembles has not been visualized. Here we present two crystal structures of this oligomerization domain. In both structures, three copies of VP35 twist about each other to form a coiled coil. This trimeric assembly is in contrast to tetrameric predictions for VP35 of Ebola virus and to known structures of homologous proteins in the measles, mumps, and Nipah viruses. Distinct oligomeric states of the Marburg and Ebola virus VP35 proteins may explain differences between them in polymerase function and immune evasion. These findings may provide a more accurate understanding of the mechanisms governing VP35's functions and inform the design of therapeutics.« less

dsRNA binding characterization of full length recombinant wild type and mutants Zaire ebolavirus VP35.

PubMed

Zinzula, Luca; Esposito, Francesca; Pala, Daniela; Tramontano, Enzo

2012-03-01

The Ebola viruses (EBOVs) VP35 protein is a multifunctional major virulence factor involved in EBOVs replication and evasion of the host immune system. EBOV VP35 is an essential component of the viral RNA polymerase, it is a key participant of the nucleocapsid assembly and it inhibits the innate immune response by antagonizing RIG-I like receptors through its dsRNA binding function and, hence, by suppressing the host type I interferon (IFN) production. Insights into the VP35 dsRNA recognition have been recently revealed by structural and functional analysis performed on its C-terminus protein. We report the biochemical characterization of the Zaire ebolavirus (ZEBOV) full-length recombinant VP35 (rVP35)-dsRNA binding function. We established a novel in vitro magnetic dsRNA binding pull down assay, determined the rVP35 optimal dsRNA binding parameters, measured the rVP35 equilibrium dissociation constant for heterologous in vitro transcribed dsRNA of different length and short synthetic dsRNA of 8bp, and validated the assay for compound screening by assessing the inhibitory ability of auryntricarboxylic acid (IC(50) value of 50μg/mL). Furthermore, we compared the dsRNA binding properties of full length wt rVP35 with those of R305A, K309A and R312A rVP35 mutants, which were previously reported to be defective in dsRNA binding-mediated IFN inhibition, showing that the latter have measurably increased K(d) values for dsRNA binding and modified migration patterns in mobility shift assays with respect to wt rVP35. Overall, these results provide the first characterization of the full-length wt and mutants VP35-dsRNA binding functions. Copyright © 2012 Elsevier B.V. All rights reserved.

Application of bluetongue Disabled Infectious Single Animal (DISA) vaccine for different serotypes by VP2 exchange or incorporation of chimeric VP2.

PubMed

Feenstra, Femke; Pap, Janny S; van Rijn, Piet A

2015-02-04

Bluetongue is a disease of ruminants caused by the bluetongue virus (BTV). Bluetongue outbreaks can be controlled by vaccination, however, currently available vaccines have several drawbacks. Further, there are at least 26 BTV serotypes, with low cross protection. A next-generation vaccine based on live-attenuated BTV without expression of non-structural proteins NS3/NS3a, named Disabled Infectious Single Animal (DISA) vaccine, was recently developed for serotype 8 by exchange of the serotype determining outer capsid protein VP2. DISA vaccines are replicating vaccines but do not cause detectable viremia, and induce serotype specific protection. Here, we exchanged VP2 of laboratory strain BTV1 for VP2 of European serotypes 2, 4, 8 and 9 using reverse genetics, without observing large effects on virus growth. Exchange of VP2 from serotype 16 and 25 was however not possible. Therefore, chimeric VP2 proteins of BTV1 containing possible immunogenic regions of these serotypes were studied. BTV1, expressing 1/16 chimeric VP2 proteins was functional in virus replication in vitro and contained neutralizing epitopes of both serotype 1 and 16. For serotype 25 this approach failed. We combined VP2 exchange with the NS3/NS3a negative phenotype in BTV1 as previously described for serotype 8 DISA vaccine. DISA vaccine with 1/16 chimeric VP2 containing amino acid region 249-398 of serotype 16 raised antibodies in sheep neutralizing both BTV1 and BTV16. This suggests that DISA vaccine could be protective for both parental serotypes present in chimeric VP2. We here demonstrate the application of the BT DISA vaccine platform for several serotypes and further extend the application for serotypes that are unsuccessful in single VP2 exchange. Copyright © 2014 Elsevier Ltd. All rights reserved.

Recombinant VP1, an Akt Inhibitor, Suppresses Progression of Hepatocellular Carcinoma by Inducing Apoptosis and Modulation of CCL2 Production

PubMed Central

Chen, Tai-An; Wang, Jui-Ling; Hung, Shao-Wen; Chu, Chiao-Li; Cheng, Yung-Chih; Liang, Shu-Mei

2011-01-01

Background The application of viral elements in tumor therapy is one facet of cancer research. Recombinant capsid protein VP1 (rVP1) of foot-and-mouth disease virus has previously been demonstrated to induce apoptosis in cancer cell lines. Here, we aim to further investigate its apoptotic mechanism and possible anti-metastatic effect in murine models of hepatocellular carcinoma (HCC), one of the most common human cancers worldwide. Methodology/Principal Findings Treatment with rVP1 inhibited cell proliferation in two murine HCC cell lines, BNL and Hepa1-6, with IC50 values in the range of 0.1–0.2 µM. rVP1 also induced apoptosis in these cells, which was mediated by Akt deactivation and dissociation of Ku70-Bax, and resulted in conformational changes and mitochondrial translocation of Bax, leading to the activation of caspases-9, -3 and -7. Treatment with 0.025 µM rVP1, which did not affect the viability of normal hepatocytes, suppressed cell migration and invasion via attenuating CCL2 production. The production of CCL2 was modulated by Akt-dependent NF-κB activation that was decreased after rVP1 treatment. The in vivo antitumor effects of rVP1 were assessed in both subcutaneous and orthotopic mouse models of HCC in immune-competent BALB/c mice. Intratumoral delivery of rVP1 inhibited subcutaneous tumor growth as a result of increased apoptosis. Intravenous administration of rVP1 in an orthotopic HCC model suppressed tumor growth, inhibited intra-hepatic metastasis, and prolonged survival. Furthermore, a decrease in the serum level of CCL2 was observed in rVP1-treated mice. Conclusions/Significance The data presented herein suggest that, via inhibiting Akt phosphorylation, rVP1 suppresses the growth, migration, and invasion of murine HCC cells by inducing apoptosis and attenuating CCL2 production both in vitro and in vivo. Recombinant protein VP1 thus has the potential to be developed as a new therapeutic agent for HCC. PMID:21826248

Evasion of interferon responses by Ebola and Marburg viruses.

PubMed

Basler, Christopher F; Amarasinghe, Gaya K

2009-09-01

The filoviruses, Ebola virus (EBOV) and Marburg virus (MARV), cause frequently lethal viral hemorrhagic fever. These infections induce potent cytokine production, yet these host responses fail to prevent systemic virus replication. Consistent with this, filoviruses have been found to encode proteins VP35 and VP24 that block host interferon (IFN)-alpha/beta production and inhibit signaling downstream of the IFN-alpha/beta and the IFN-gamma receptors, respectively. VP35, which is a component of the viral nucleocapsid complex and plays an essential role in viral RNA synthesis, acts as a pseudosubstrate for the cellular kinases IKK-epsilon and TBK-1, which phosphorylate and activate interferon regulatory factor 3 (IRF-3) and interferon regulatory factor 7 (IRF-7). VP35 also promotes SUMOylation of IRF-7, repressing IFN gene transcription. In addition, VP35 is a dsRNA-binding protein, and mutations that disrupt dsRNA binding impair VP35 IFN-antagonist activity while leaving its RNA replication functions intact. The phenotypes of recombinant EBOV bearing mutant VP35s unable to inhibit IFN-alpha/beta demonstrate that VP35 IFN-antagonist activity is critical for full virulence of these lethal pathogens. The structure of the VP35 dsRNA-binding domain, which has recently become available, is expected to provide insight into how VP35 IFN-antagonist and dsRNA-binding functions are related. The EBOV VP24 protein inhibits IFN signaling through an interaction with select host cell karyopherin-alpha proteins, preventing the nuclear import of otherwise activated STAT1. It remains to be determined to what extent VP24 may also modulate the nuclear import of other host cell factors and to what extent this may influence the outcome of infection. Notably, the Marburg virus VP24 protein does not detectably block STAT1 nuclear import, and, unlike EBOV, MARV infection inhibits STAT1 and STAT2 phosphorylation. Thus, despite their similarities, there are fundamental differences by which these deadly viruses counteract the IFN system. It will be of interest to determine how these differences influence pathogenesis.

A Single Amino Acid Change in the Marburg Virus Matrix Protein VP40 Provides a Replicative Advantage in a Species-Specific Manner

PubMed Central

Koehler, Alexander; Kolesnikova, Larissa; Welzel, Ulla; Schudt, Gordian; Herwig, Astrid

2015-01-01

ABSTRACT Marburg virus (MARV) induces severe hemorrhagic fever in humans and nonhuman primates but only transient nonlethal disease in rodents. However, sequential passages of MARV in rodents boosts infection leading to lethal disease. Guinea pig-adapted MARV contains one mutation in the viral matrix protein VP40 at position 184 (VP40D184N). The contribution of the D184N mutation to the efficacy of replication in a new host is unknown. In the present study, we demonstrated that recombinant MARV containing the D184N mutation in VP40 [rMARVVP40(D184N)] grew to higher titers than wild-type recombinant MARV (rMARVWT) in guinea pig cells. Moreover, rMARVVP40(D184N) displayed higher infectivity in guinea pig cells. Comparative analysis of VP40 functions indicated that neither the interferon (IFN)-antagonistic function nor the membrane binding capabilities of VP40 were affected by the D184N mutation. However, the production of VP40-induced virus-like particles (VLPs) and the recruitment of other viral proteins to the budding site was improved by the D184N mutation in guinea pig cells, which resulted in the higher infectivity of VP40D184N-induced infectious VLPs (iVLPs) compared to that of VP40-induced iVLPs. In addition, the function of VP40 in suppressing viral RNA synthesis was influenced by the D184N mutation specifically in guinea pig cells, thus allowing greater rates of transcription and replication. Our results showed that the improved viral fitness of rMARVVP40(D184N) in guinea pig cells was due to the better viral assembly function of VP40D184N and its lower inhibitory effect on viral transcription and replication rather than modulation of the VP40-mediated suppression of IFN signaling. IMPORTANCE The increased virulence achieved by virus passaging in a new host was accompanied by mutations in the viral genome. Analyzing how these mutations affect the functions of viral proteins and the ability of the virus to grow within new host cells helps in the understanding of the molecular mechanisms increasing virulence. Using a reverse genetics approach, we demonstrated that a single mutation in MARV VP40 detected in a guinea pig-adapted MARV provided a replicative advantage of rMARVVP40(D184N) in guinea pig cells. Our studies show that this replicative advantage of rMARV VP40D184N was based on the improved functions of VP40 in iVLP assembly and in the regulation of transcription and replication rather than on the ability of VP40 to combat the host innate immunity. PMID:26581998