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1

Human Respiratory Syncytial Virus Memphis 37 Grown in HEp-2 Cells Causes more Severe Disease in Lambs than Virus Grown in Vero Cells  

PubMed Central

Respiratory syncytial virus (RSV) is the most common cause of bronchiolitis in infants and young children. A small percentage of these individuals develop severe and even fatal disease. To better understand the pathogenesis of severe disease and develop therapies unique to the less-developed infant immune system, a model of infant disease is needed. The neonatal lamb pulmonary development and physiology is similar to that of infants, and sheep are susceptible to ovine, bovine, or human strains of RSV. RSV grown in Vero (African green monkey) cells has a truncated attachment G glycoprotein as compared to that grown in HEp-2 cells. We hypothesized that the virus grown in HEp-2 cells would cause more severe clinical symptoms and cause more severe pathology. To confirm the hypothesis, lambs were inoculated simultaneously by two different delivery methods (intranasal and nebulized inoculation) with either Vero-grown or HEp-2-grown RSV Memphis 37 (M37) strain of virus to compare viral infection and disease symptoms. Lambs infected with HEp-2 cell-derived virus by either intranasal or nebulization inoculation had significantly higher levels of viral RNA in lungs as well as greater clinical disease including both gross and histopathologic lesions compared to lambs similarly inoculated with Vero-grown virus. Thus, our results provide convincing in vivo evidence for differences in viral infectivity that corroborate previous in vitro mechanistic studies demonstrating differences in the G glycoprotein expression by RSV grown in Vero cells.

Derscheid, Rachel J.; van Geelen, Albert; McGill, Jodi L.; Gallup, Jack M.; Cihlar, Tomas; Sacco, Randy E.; Ackermann, Mark R.

2013-01-01

2

Human respiratory syncytial virus Memphis 37 grown in HEp-2 cells causes more severe disease in lambs than virus grown in Vero cells.  

PubMed

Respiratory syncytial virus (RSV) is the most common cause of bronchiolitis in infants and young children. A small percentage of these individuals develop severe and even fatal disease. To better understand the pathogenesis of severe disease and develop therapies unique to the less-developed infant immune system, a model of infant disease is needed. The neonatal lamb pulmonary development and physiology is similar to that of infants, and sheep are susceptible to ovine, bovine, or human strains of RSV. RSV grown in Vero (African green monkey) cells has a truncated attachment G glycoprotein as compared to that grown in HEp-2 cells. We hypothesized that the virus grown in HEp-2 cells would cause more severe clinical symptoms and cause more severe pathology. To confirm the hypothesis, lambs were inoculated simultaneously by two different delivery methods (intranasal and nebulized inoculation) with either Vero-grown or HEp-2-grown RSV Memphis 37 (M37) strain of virus to compare viral infection and disease symptoms. Lambs infected with HEp-2 cell-derived virus by either intranasal or nebulization inoculation had significantly higher levels of viral RNA in lungs as well as greater clinical disease including both gross and histopathologic lesions compared to lambs similarly inoculated with Vero-grown virus. Thus, our results provide convincing in vivo evidence for differences in viral infectivity that corroborate previous in vitro mechanistic studies demonstrating differences in the G glycoprotein expression by RSV grown in Vero cells. PMID:24284879

Derscheid, Rachel J; van Geelen, Albert; McGill, Jodi L; Gallup, Jack M; Cihlar, Tomas; Sacco, Randy E; Ackermann, Mark R

2013-11-01

3

A microcarrier cell culture process for propagating rabies virus in Vero cells grown in a stirred bioreactor under fully animal component free conditions  

Microsoft Academic Search

Rabies virus strain production in Vero cells grown on Cytodex 1 in a 2L stirred tank bioreactor and in a medium free of components of human or animal origin (VP-SFM) is described. Cell banking procedure in VP-SFM supplemented with an animal components free mixture (10%DMSO+0.1%methylcellulose) was reported and cell growth after revitalization was assessed. Vero cells exhibited growth performances (specific

Samia Rourou; Arno van der Ark; Tiny van der Velden; Héla Kallel

2007-01-01

4

A microcarrier cell culture process for propagating rabies virus in Vero cells grown in a stirred bioreactor under fully animal component free conditions.  

PubMed

Rabies virus strain production in Vero cells grown on Cytodex 1 in a 2 L stirred tank bioreactor and in a medium free of components of human or animal origin (VP-SFM) is described. Cell banking procedure in VP-SFM supplemented with an animal components free mixture (10%DMSO+0.1%methylcellulose) was reported and cell growth after revitalization was assessed. Vero cells exhibited growth performances (specific growth rate and cell division number) similar to that obtained in serum containing medium. To design a scalable process that is totally free of animal-derived substances, two proteases: TrypLE Select and Accutase, were assessed as an alternative to trypsin which is routinely used for cell passage. Growth performance of Vero cells grown in VP-SFM and MEM+10% fetal calf serum (FCS) over four passages and subcultivated with either TrypLE Select or Accutase was evaluated. TrypLE Select showed the best performance in terms of specific growth rate and cell division number. Kinetics of cell growth and rabies virus production (LP2061/Vero strain) were investigated in spinner flask and in a 2 L bioreactor. In spinner flask, a maximal cell density level of 1.85x10(6) cells/mL was achieved when the cells were grown in VP-SFM on 2 g/L Cytodex 1. Cell infection experiments conducted at an MOI of 0.3 and without the medium exchange step, typically needed for serum containing rabies virus production, resulted in a maximal virus titer equal to 2x10(7) (Fluorescent Focus Unit) FFU/mL. In stirred tank bioreactor, Vero cell growth in VP-SFM on 3 g/L Cytodex 1 was shown to be sensitive to the aeration mode. Sparging the culture was detrimental for cell growth, whereas cell density level was greatly enhanced when only headspace aeration was used. A cell density level of 2.6x10(6) cells/mL was obtained when the cells were grown on 3g/L Cytodex 1 and in batch culture mode. Cell infection at an MOI of 0.1 without any medium exchange, yielded a maximal rabies virus titer of 2.4x10(7) FFU/mL. Furthermore, Vero cell growth in a 2 L bioreactor using recirculation culture mode during cell proliferation step and perfusion for virus multiplication phase was investigated. In comparison to batch culture, a higher cell density level that was equal to 5x10(6) cells/mL was reached. Cell infection under conditions similar to batch culture, resulted in a maximal virus titer equal to 1.38x10(8) FFU/mL. The potency of the pooled inactivated virus harvests showed an activity of 2.58 IU/mL which was comparable to that obtained in serum supplemented medium. PMID:17307281

Rourou, Samia; van der Ark, Arno; van der Velden, Tiny; Kallel, Héla

2007-05-10

5

Purification and Characterization of Enterovirus 71 Viral Particles Produced from Vero Cells Grown in a Serum-Free Microcarrier Bioreactor System  

Microsoft Academic Search

BackgroundEnterovirus 71 (EV71) infections manifest most commonly as a childhood exanthema known as hand-foot-and-mouth disease (HFMD) and can cause neurological disease during acute infection.Principal FindingIn this study, we describe the production, purification and characterization of EV71 virus produced from Vero cells grown in a five-liter serum-free bioreactor system containing 5 g\\/L Cytodex 1 microcarrier. The viral titer was >106 TCID50\\/mL

Chia-Chyi Liu; Meng-Shin Guo; Fion Hsiao-Yu Lin; Kuang-Nan Hsiao; Kate Hsuen-Wen Chang; Ai-Hsiang Chou; Yu-Chao Wang; Yu-Ching Chen; Chung-Shi Yang; Pele Choi-Sing Chong

2011-01-01

6

A novel animal-component-free medium for rabies virus production in Vero cells grown on Cytodex 1 microcarriers in a stirred bioreactor  

Microsoft Academic Search

Vero cells growth and rabies production in IPT-AF medium, a property animal-component-free medium are described in this work.\\u000a Kinetics of cell growth and rabies virus (strain LP 2061) production were first conducted in spinner flasks. Over eight independent\\u000a experiments, Vero cell growth in IPT-AF medium, on 2 g\\/l Cytodex 1 was consistent. An average Cd (cell division number) of\\u000a 3.3?±?0.4 and

Samia Rourou; Arno van der Ark; Samy Majoul; Khaled Trabelsi; Tiny van der Velden; Héla Kallel

2009-01-01

7

Purification and Characterization of Enterovirus 71 Viral Particles Produced from Vero Cells Grown in a Serum-Free Microcarrier Bioreactor System  

PubMed Central

Background Enterovirus 71 (EV71) infections manifest most commonly as a childhood exanthema known as hand-foot-and-mouth disease (HFMD) and can cause neurological disease during acute infection. Principal Finding In this study, we describe the production, purification and characterization of EV71 virus produced from Vero cells grown in a five-liter serum-free bioreactor system containing 5 g/L Cytodex 1 microcarrier. The viral titer was >106 TCID50/mL by 6 days post infection when a MOI of 10?5 was used at the initial infection. Two EV71 virus fractions were separated and detected when the harvested EV71 virus concentrate was purified by sucrose gradient zonal ultracentrifugation. The EV71 viral particles detected in the 24–28% sucrose fractions had an icosahedral structure 30–31 nm in diameter and had low viral infectivity and RNA content. Three major viral proteins (VP0, VP1 and VP3) were observed by SDS-PAGE. The EV71 viral particles detected in the fractions containing 35–38% sucrose were 33–35 nm in size, had high viral infectivity and RNA content, and were composed of four viral proteins (VP1, VP2, VP3 and VP4), as shown by SDS-PAGE analyses. The two virus fractions were formalin-inactivated and induced high virus neutralizing antibody responses in mouse immunogenicity studies. Both mouse antisera recognized the immunodominant linear neutralization epitope of VP1 (residues 211–225). Conclusion These results provide important information for cell-based EV71 vaccine development, particularly for the preparation of working standards for viral antigen quantification.

Liu, Chia-Chyi; Guo, Meng-Shin; Lin, Fion Hsiao-Yu; Hsiao, Kuang-Nan; Chang, Kate Hsuen-Wen; Chou, Ai-Hsiang; Wang, Yu-Chao; Chen, Yu-Ching; Yang, Chung-Shi; Chong, Pele Choi-Sing

2011-01-01

8

Kinetics and metabolic specificities of Vero cells in bioreactor cultures with serum-free medium  

Microsoft Academic Search

The aim of this study was to understand the metabolism kinetics of Vero cells grown on microcarriers in bioreactors in serum-free\\u000a medium (SFM). We sought to determine what nutrients are essential for Vero cells and how they are consumed. Contrary to glucose\\u000a and to most of the amino acids, glutamine and serine were very quickly depleted in this medium and

Sébastien Quesney; Annie Marc; Catherine Gerdil; Cyrille Gimenez; Jacqueline Marvel; Yves Richard; Bernard Meignier

2003-01-01

9

African green monkey kidney (Vero) cells provide an alternative host cell system for influenza A and B viruses.  

PubMed

The preparation of live, attenuated human influenza virus vaccines and of large quantities of inactivated vaccines after the emergence or reemergence of a pandemic influenza virus will require an alternative host cell system, because embryonated chicken eggs will likely be insufficient and suboptimal. Preliminary studies indicated that an African green monkey kidney cell line (Vero) is a suitable system for the primary isolation and cultivation of influenza A viruses (E. A. Govorkova, N. V. Kaverin, L. V. Gubareva, B. Meignier, and R. G. Webster, J. Infect. Dis. 172:250-253, 1995). We now demonstrate for the first time that Vero cells are suitable for isolation and productive replication of influenza B viruses and determine the biological and genetic properties of both influenza A and B viruses in Vero cells; additionally, we characterize the receptors on Vero cells compared with those on Madin-Darby canine kidney (MDCK) cells. Sequence analysis indicated that the hemagglutinin of Vero cell-derived influenza B viruses was identical to that of MDCK-grown counterparts but differed from that of egg-grown viruses at amino acid positions 196 to 198. Fluorescence-activated cell sorting analysis showed that although Vero cells possess predominantly alpha2,3 galactose-linked sialic acid, they are fully susceptible to infection with either human influenza A or B viruses. Moreover, all virus-specific polypeptides were synthesized in the same proportions in Vero cells as in MDCK cells. Electron microscopic and immunofluorescence studies confirmed that infected Vero cells undergo the same morphological changes as do other polarized epithelia] cells. Taken together, these results indicate that Vero cell lines could serve as an alternative host system for the cultivation of influenza A and B viruses, providing adequate quantities of either virus to meet the vaccine requirements imposed by an emerging pandemic. PMID:8764064

Govorkova, E A; Murti, G; Meignier, B; de Taisne, C; Webster, R G

1996-08-01

10

Recycle of Cytodex-3 in Vero cell culture  

Microsoft Academic Search

It is commonly considered not desirable to use microcarriers more than once in the cultivation of anchorage-dependent animal cells. However, our experiment contradicts this belief. The collagen-coated microcarriers, Cytodex-3, from a batch culture of Vero cells, were collected, cooled to 4, agitated in basic phosphate-buffered solution to detach the cells, and then fully washed to remove the cell debris. The

Y. Wang; F. Ouyang

1999-01-01

11

Optimization of Toxoplasma gondii cultivation in VERO cell line.  

PubMed

In vitro culture of Toxoplasma gondii can provide tachyzoites which are active, viable and with desirable purity. Thus the aim of this study was to optimize the cell culture method for T. gondii propagation to obtain a consistent source of parasites with maximum yield and viability, but minimum host cell contamination for use in production of excretory-secretory antigen. Tachyzoites with seed counts of 1x10(6), 1x10(7) and 1x10(8) harvested from infected mice were added to VERO cells of different degrees of confluence, namely 50%, 85% and 100%, and examined periodically using an inverted microscope. When the maximum release of the tachyzoites was observed from the host cells, the culture supernatant was removed and the tachyzoites harvested. Using a Neubauer chamber, the percentages of viable tachyzoites and host cell contamination were determined using trypan blue stain. Parameters that gave the best yield and purity of viable tachyzoites were found to be as follows: VERO cells at 85% confluence in DMEM medium and inoculum comprising 1x10(7) tachyzoites. After about 3 days post infection, the tachyzoites multiplied 78x, with a yield of ~7.8x10(8) per flask, 99% viability and 3% host cell contamination. This study has successfully optimized the method of propagation of T. gondii tachyzoites in VERO cells which produce parasites with high yield, purity and viability. PMID:20562822

Saadatnia, G; Haj Ghani, H; Khoo, B Y; Maimunah, A; Rahmah, N

2010-04-01

12

Establishment of a Vero cell line persistently infected with African swine fever virus.  

PubMed Central

A Vero cell line persistently infected with African swine fever virus was established by infecting the cells in the presence of 10 mM NH4Cl (Vero-P cell line). The virus derived from the Vero-P cultures infected Vero cells, and virus titers were comparable to those obtained in Vero cells acutely infected with African swine fever virus. The structural proteins of the virus from Vero-P cells were similar to those of the virus produced in lytic infections. Virus production was low when the Vero-P cells were growing logarithmically and increased considerably in confluent cultures when lysis appeared in a fraction of the cell population. Images

Salas, J; Vinuela, E

1986-01-01

13

Development of Vero cell-derived inactivated Japanese encephalitis vaccine.  

PubMed

We have established a manufacturing system for a Vero cell-derived inactivated Japanese encephalitis vaccine at a 500l scale. The production system involves expansion of Vero cells using microcarrier, followed by virus infection. Except for an additional purification step, the downstream purification processes are similar to those used for the current mouse brain-derived vaccine; cell removal, concentration and removal of low-molecular weight impurities by membrane filtration, formalin-inactivation, sucrose density gradient ultracentrifugation, and Sulfate-Cellulofine column chromatography are conducted. The antigen obtained from the manufacturing system was highly purified and its physico-chemical and immunological properties were comparable with those of antigen derived from mouse brains. Our system is very simple and could be easily scaled-up to allow vaccine production at a several thousand litre scale. PMID:12421588

Sugawara, Keishin; Nishiyama, Kiyoto; Ishikawa, Yuji; Abe, Motoharu; Sonoda, Kengo; Komatsu, Kazuhiro; Horikawa, Yoshikane; Takeda, Kengo; Honda, Tomitaka; Kuzuhara, Shoji; Kino, Yoichiro; Mizokami, Hiroshi; Mizuno, Kyosuke; Oka, Tetsuya; Honda, Kennosuke

2002-12-01

14

Improved poliovirus d-antigen yields by application of different Vero cell cultivation methods.  

PubMed

Vero cells were grown adherent to microcarriers (Cytodex 1; 3gL(-1)) using animal component free media in stirred-tank type bioreactors. Different strategies for media refreshment, daily media replacement (semi-batch), continuous media replacement (perfusion) and recirculation of media, were compared with batch cultivation. Cell densities increased using a feed strategy from 1×10(6) cellsmL(-1) during batch cultivation to 1.8, 2.7 and 5.0×10(6) cellsmL(-1) during semi-batch, perfusion and recirculation, respectively. The effects of these different cell culture strategies on subsequent poliovirus production were investigated. Increased cell densities allowed up to 3 times higher d-antigen levels when compared with that obtained from batch-wise Vero cell culture. However, the cell specific d-antigen production was lower when cells were infected at higher cell densities. This cell density effect is in good agreement with observations for different cell lines and virus types. From the evaluated alternative culture methods, application of a semi-batch mode of operations allowed the highest cell specific d-antigen production. The increased product yields that can easily be reached using these higher cell density cultivation methods, showed the possibility for better use of bioreactor capacity for the manufacturing of polio vaccines to ultimately reduce vaccine cost per dose. Further, the use of animal-component-free cell- and virus culture media shows opportunities for modernization of human viral vaccine manufacturing. PMID:24583004

Thomassen, Yvonne E; Rubingh, Olaf; Wijffels, René H; van der Pol, Leo A; Bakker, Wilfried A M

2014-05-19

15

VERO cells (cercopithecus aethiops kidney) — growth characteristics and viral susceptibility for use in diagnostic virology  

Microsoft Academic Search

Summary An investigation of some of the characteristics of the VERO cell line (Cercopithecus aethiops kidney) is reported, in which the suitability of the cells for use in routine diagnostic virology was examined. VERO cells will:1.grow to monolayers as rapidly as other heteroploid cell lines, but will maintain as usable monolayers in conventional maintenance medium for a significantly longer time;2.grow

D. E. Macfarlane; R. G. Sommerville

1969-01-01

16

Decreased sensitivity to diphtheria toxin of Vero cells cultured in serum-free medium  

Microsoft Academic Search

Vero cell cultures are used in the quality control of Diphtheria vaccines: to estimate vaccine potency and to determine residual toxicity and reversion to toxicity. The impact of replacing foetal calf serum containing medium (SCM) by serum free media (SFM) on the sensitivity of Vero cells to Diphtheria Toxin was studied. Compared to SCM, SFM showed an eight-fold decrease in

S. J. Piersma; J. W. van der Gun; C. F. M. Hendriksen; M. Thalen

2005-01-01

17

Live cold-adapted influenza A vaccine produced in Vero cell line  

Microsoft Academic Search

The African green monkey kidney (Vero) cell line was used as a substrate for the development of a live cold-adapted (ca) reassortant influenza vaccine. For that purpose, a new master strain was generated by an adaptation of the wild type (wt) A\\/Singapore\\/1\\/57 virus to growth at 25°C in a Vero cell line. The resulting cold-adapted (ca) muster strain A\\/Singapore\\/1\\/57ca showed

Julia Romanova; Dietmar Katinger; Boris Ferko; Brigitta Vcelar; Sabine Sereinig; Oleg Kuznetsov; Marina Stukova; Marjana Erofeeva; Oleg Kiselev; Hermann Katinger; Andrej Egorov

2004-01-01

18

Humoral and cell-mediated immunity to vero cell-derived influenza vaccine.  

PubMed

A candidate trivalent influenza whole virus vaccine produced in a continuous mammalian cell line (Vero), and analogous commercially available egg-derived vaccines, were compared for their ability to induce humoral and cell-mediated immunity in Balb/c mice. Substantial haemagglutination-inhibition titre and high levels of influenza virus-specific IgG were found in all groups of immunized mice, irrespective of the vaccine formulation. The IgG responses were predominantly of IgG1 and IgG2a/2b isotypes. Virus-specific secretory IgA antibodies were detected only in mice immunized intranasally with a live virus, derived either from Vero cells or eggs. T-cell proliferative responses and T-helper 1 type cytokine release was significantly higher in mice immunized with Vero cell-derived influenza vaccine compared to egg-derived vaccine formulations. We have demonstrated that the immunogenicity of the trivalent Vero cell-derived whole influenza virus vaccine was comparable to that of the equivalent egg-derived vaccine, with respect to humoral immune response and was superior with respect to cellular response. PMID:11137251

Brühl, P; Kerschbaum, A; Kistner, O; Barrett, N; Dorner, F; Gerencer, M

2000-12-01

19

Changes in antiviral susceptibility to entry inhibitors and endocytic uptake of dengue-2 virus serially passaged in Vero or C6/36 cells.  

PubMed

The aim of the present study was to analyze the influence of virus origin, mammalian or mosquito cell-derived, on antiviral susceptibility of DENV-2 to entry inhibitors and the association of this effect with any alteration in the mode of entry into the cell. To this end, ten serial passages of DENV-2 were performed in mosquito C6/36 cells or monkey Vero cells and the antiviral susceptibility of each virus passage to sulfated polysaccharides (SPs), like heparin and carrageenans, was evaluated by a virus plaque reduction assay. After serial passaging in Vero cells, DENV-2 became increasingly resistant to SP inhibition whereas the antiviral susceptibility was not altered in virus propagated in C6/36 cells. The change in antiviral susceptibility was associated to a differential mode of entry into the host cell. The route of endocytic entry for productive Vero cell infection was altered from a non-classical clathrin independent pathway for C6/36-grown virus to a clathrin-mediated endocytosis when the virus was serially propagated in Vero cells. Our results show the impact of the cellular system used for successive propagation of DENV on the initial interaction between the host cell and the virion in the next round of infection and the relevant consequences it might have during the in vitro evaluation of entry inhibitors. PMID:24583230

Acosta, Eliana G; Piccini, Luana E; Talarico, Laura B; Castilla, Viviana; Damonte, Elsa B

2014-05-12

20

Humoral and cell-mediated immunity to Vero cell-derived influenza vaccine  

Microsoft Academic Search

A candidate trivalent influenza whole virus vaccine produced in a continuous mammalian cell line (Vero), and analogous commercially available egg-derived vaccines, were compared for their ability to induce humoral and cell-mediated immunity in Balb\\/c mice. Substantial haemagglutination-inhibition titre and high levels of influenza virus-specific IgG were found in all groups of immunized mice, irrespective of the vaccine formulation. The IgG

Peter Brühl; Astrid Kerschbaum; Otfried Kistner; Noel Barrett; Friedrich Dorner; Marijan Geren?er

2000-01-01

21

Vero cell culture-derived pandemic influenza vaccines: preclinical and clinical development.  

PubMed

Several subtypes of influenza A viruses with pandemic potential are endemic in bird populations throughout Asia, Africa and the Middle East, and evidence suggests that these viruses are adapting to the mammalian host. As emphasized by the high mortality rate of humans infected with H5N1 viruses, this situation presents a substantial risk to global human health. The Vero cell culture platform has been used to develop whole-virus influenza vaccines that provide broad cross-clade protection against viruses with pandemic potential, at low antigen doses, without the requirement for adjuvantation. The safety and immunogenicity of these vaccines has been demonstrated in studies with more than 10,000 individuals, including healthy adult and elderly subjects, children and various risk groups. These Vero cell-derived vaccines are licensed for prepandemic and pandemic use. The Vero platform is also being explored to develop next-generation live-attenuated and recombinant vaccines. PMID:23560920

Barrett, P Noel; Portsmouth, Daniel; Ehrlich, Hartmut J

2013-04-01

22

Preflucel®: a Vero-cell culture-derived trivalent influenza vaccine.  

PubMed

Vaccination is the principal means to reduce the impact of influenza infection. Effective vaccination programs require a reliable and safe production system. Traditionally, influenza vaccines are produced in embryonated chicken eggs. Over the last two decades, new cell culture-derived vaccines have been licensed and manufactured, and other vaccines are still in various phases of development. Vero cells have been used for the development of a wide variety of vaccines including influenza vaccines. Pandemic and avian influenza vaccines derived from Vero cells have been shown to be well tolerated and immunogenic in animal and Phase I-II clinical studies. A Phase III randomized, double-blind, placebo-controlled trial of a trivalent influenza vaccine produced in Vero-cell culture was conducted in 7250 adults aged 18-49 years. Overall protective efficacy for antigenically matched influenza vaccine was 78.5%. The vaccine was well tolerated with no treatment-related serious adverse events and compared favorably with egg-derived vaccines from previous trials. Vero-cell-derived influenza vaccines have the potential to be an important parts of the influenza vaccine strategy, especially if an avian-derived strain becomes predominant or the demand outstrips the capacity of egg-based production systems. PMID:22913252

Chan, Candice Yuen-Yue; Tambyah, Paul Anantharajah

2012-07-01

23

Polyvinyl formal surface promotes continuous growth of Vero cells in protein-free medium.  

PubMed

The effect of polyvinyl formal (PVF) culture surface on the growth of 10 mammalian continuous cell lines, including Swiss 3T6, NCTC clone 929 L, BHK-21 clone 13, CHO-K1, PK 15, A 431, HeLa, MDCK, LLC-MK2 and Vero in protein-free 1:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12 supplemented with trace elements and L-ascorbic acid 2-phosphate, was investigated. Most of the cell lines showed only some initial proliferation on PVF similar to the polystyrene (PS) surface of commercially available culture flasks. In contrast, proliferation of monkey kidney cell line Vero was by far greater on PVF than on PS or poly-D-lysine treated culture surface. In addition, Vero cells on PVF could be subcultured in the protein-free medium without any significant decrease of growth rate in successive passages. These results showed that PVF provides a culture surface which selectively promotes continuous growth of Vero cells in protein-free, chemically defined medium. PMID:1610559

Cinatl, J; Cinatl, J; Rabenau, H; Doerr, H W

1992-03-01

24

Development of a mammalian cell (Vero) derived candidate influenza virus vaccine  

Microsoft Academic Search

Influenza vaccine production is dependent on the availability of embryonated hen eggs for virus growth. This is an extremely cumbersome system with many disadvantages with respect to selection of virus variants and presence of adventitious viruses. We have developed an alternative cell culture system which allows rapid production of large volumes of vaccine. The World Health Organisation (WHO) approved Vero

O Kistner; P. N. Barrett; W. Mundt; M. Reiter; S. Schober-Bendixen; F. Dorner

1998-01-01

25

A purified inactivated Japanese encephalitis virus vaccine made in vero cells  

Microsoft Academic Search

A second generation, purified, inactivated vaccine (PIV) against Japanese encephalitis (JE) virus was produced and tested in mice where it was found to be highly immunogenic and protective. The JE-PIV was made from an attenuated strain of JE virus propagated in certified Vero cells, purified, and inactivated with formalin. Its manufacture followed current GMP guidelines for the production of biologicals.

Ashok K. Srivastava; J. Robert Putnak; Sung H. Lee; Sun P. Hong; Sang B. Moon; David A. Barvir; Bangti Zhao; Russell A. Olson; Soo-Ok Kim; Wang-Don Yoo; Andrew C. Towle; David W. Vaughn; Bruce L. Innis; Kenneth H. Eckels

2001-01-01

26

Japanese encephalitis virus production in Vero cells with serum-free medium using a novel oscillating bioreactor  

Microsoft Academic Search

A novel oscillating bioreactor, BelloCell, was successfully applied for the cultivation of Vero cells using serum-free medium, and the production of Japanese encephalitis virus. The BelloCell requires no air sparging, pumping, or agitation, and thus provides a low shear environment. Owing to its simple design, BelloCell is extremely easy to handle and operate. Using this BelloCell (500ml culture), Vero cells

Hiroko Toriniwa; Tomoyoshi Komiya

2007-01-01

27

Vero cell platform in vaccine production: moving towards cell culture-based viral vaccines.  

PubMed

The development of cell culture systems for virus propagation has led to major advances in virus vaccine development. Primary and diploid cell culture systems are now being replaced by the use of continuous cell lines (CCLs). These substrates are gaining increasing acceptance from regulatory authorities as improved screening technologies remove fears regarding their potential oncogenic properties. The Vero cell line is the most widely accepted CCL by regulatory authorities and has been used for over 30 years for the production of polio and rabies virus vaccines. The recent licensure of a Vero cell-derived live virus vaccine (ACAM2000, smallpox vaccine) has coincided with an explosion in the development of a range of new viral vaccines, ranging from live-attenuated pediatric vaccines against rotavirus infections to inactivated whole-virus vaccines against H5N1 pandemic influenza. These developments have illustrated the value of this cell culture platform in the rapid development of vaccines against a range of virus diseases. PMID:19397417

Barrett, P Noel; Mundt, Wolfgang; Kistner, Otfried; Howard, M Keith

2009-05-01

28

Differential activities of plant polyphenols on the binding and internalization of cholera toxin in vero cells.  

PubMed

Plant polyphenols, RG-tannin, and applephenon had been reported to inhibit cholera toxin (CT) ADP-ribosyltransferase activity and CT-induced fluid accumulation in mouse ileal loops. A high molecular weight fraction of hop bract extract (HBT) also inhibited CT ADP-ribosyltransferase activity. We report here the effect of those polyphenols on the binding and entry of CT into Vero cells. Binding of CT to Vero cells or to ganglioside GM1, a CT receptor, was inhibited in a concentration-dependent manner by HBT and applephenon but not RG-tannin. These observations were confirmed by fluorescence microscopy using Cy3-labeled CT. Following toxin binding to cells, applephenon, HBT, and RG-tannin suppressed its internalization. HBT or applephenon precipitated CT, CTA, and CTB from solution, creating aggregates larger than 250 kDa. In contrast, RG-tannin precipitated CT poorly; it formed complexes with CT, CTA, or CTB, which were demonstrated with sucrose density gradient centrifugation and molecular weight exclusion filters. In agreement, CTA blocked the inhibition of CT internalization by RG-tannin. These data suggest that some plant polyphenols, similar to applephenon and HBT, bind CT, forming large aggregates in solution or, perhaps, on the cell surface and thereby suppress CT binding and internalization. In contrast, RG-tannin binding to CT did not interfere with its binding to Vero cells or GM1, but it did inhibit internalization. PMID:15814610

Morinaga, Naoko; Iwamaru, Yoshifumi; Yahiro, Kinnosuke; Tagashira, Motoyuki; Moss, Joel; Noda, Masatoshi

2005-06-17

29

Decreased sensitivity to diphtheria toxin of Vero cells cultured in serum-free medium.  

PubMed

Vero cell cultures are used in the quality control of Diphtheria vaccines: to estimate vaccine potency and to determine residual toxicity and reversion to toxicity. The impact of replacing foetal calf serum containing medium (SCM) by serum free media (SFM) on the sensitivity of Vero cells to Diphtheria Toxin was studied. Compared to SCM, SFM showed an eight-fold decrease in sensitivity to Diphtheria Toxin. This decrease was almost immediate, indicating that this phenomenon was not caused by a change in membrane structure or protein expression. We investigated the effect of SFM on Diphtheria Toxin in order to determine the cause of the decrease in sensitivity. Our results show that oligopeptides, which are often used in SFM as part of the replacement of foetal calf serum, are the most likely cause. PMID:15905099

Piersma, S J; van der Gun, J W; Hendriksen, C F M; Thalen, M

2005-06-01

30

Structural and functional analysis of virus factories purified from Rabbit vesivirus-infected Vero cells  

Microsoft Academic Search

Rabbit vesivirus infection induces membrane modifications and accumulation of vesicular structures in the cytoplasm of infected Vero cells. Crude RaV replication complexes (RCs) have been purified and their structural and functional properties have been characterized. We show that calnexin, an ER-resident protein, RaV non-structural proteins 2AB-, 2C-, 3A-, 3B- and 3CD-like as well as viral RNAs co-localize within membranous structures

Rosa Casais; Lorenzo González Molleda; Angeles Machín; Gloria del Barrio; Alberto García Manso; Kevin P. Dalton; Ana Coto; José Manuel Martín Alonso; Miguel Prieto; Francisco Parra

2008-01-01

31

Isolation of Mycoplasma genitalium from First-Void Urine Specimens by Coculture with Vero Cells?  

PubMed Central

Isolation of Mycoplasma genitalium from clinical specimens remains difficult. We describe an improvement of the Vero cell coculture method in which the growth of M. genitalium was monitored by quantitative real-time PCR. Four new M. genitalium strains were isolated from six first-void urine specimens of male Japanese patients with urethritis. In two of them, only M. genitalium was detected: one also contained Ureaplasma urealyticum, and one contained Chlamydia trachomatis, Neisseria gonorrhoeae, U. urealyticum, and Ureaplasma parvum. In the specimens yielding isolates of M. genitalium, growth was documented by quantitative PCR after two to five passages in Vero cells. The complete isolation procedure from the initial inoculation to completion of single-colony cloning took about 1 year. Isolation of M. genitalium from urine specimens proved to be more difficult than from swab specimens. Due to the cytotoxic effect of urine, a procedure involving washing of the urinary sediment was introduced. Furthermore, prolonged storage of the urine specimens before culture was shown to be detrimental to the success of isolation, as shown by the lack of success in attempts to isolate M. genitalium from mailed urine specimens as well as by simulation experiments. High concentrations of penicillin G and amphotericin B were surprisingly inhibitory to the growth of wild-type M. genitalium strains, but penicillin G at 200 IU/ml and polymyxin B at 500 ?g/ml could be used as selective antibiotics to avoid bacterial overgrowth in the Vero cell cultures.

Hamasuna, Ryoichi; Osada, Yukio; Jensen, J?rgen Skov

2007-01-01

32

Optimization of transfection methods for Huh-7 and Vero cells: comparative study.  

PubMed

Availability of an efficient transfection protocol is the first determinant in success of gene transferring studies in mammalian cells which is accomplished experimentally for every single cell type. Herein, we provide data of a comparative study on optimization of transfection condition by electroporation and chemical methods for Huh-7 and Vero cells. Different cell confluencies, DNA/reagent ratios and total transfection volumes were optimized for two chemical reagents including jetPEI and Lipofectamine 2000. Besides, the effects of electric field strength and pulse length were investigated to improve electroporation efficiency. Transfection of cells by pEGFP-N1 vector and tracking the expression of GFP by FACS and Fluorescence Microscopy analysis were the employed methods to evaluate transfection efficiencies. Optimized electroporation protocols yielded 63.73 +/- 2.36 and 73.9 +/- 1.6% of transfection in Huh-7 and Vero cells respectively, while maximum achieved level of transfection by jetPEI was respectively 14.2 +/- 0.69 and 28 +/- 1.11% for the same cells. Post transfectional chilling of the cells did not improve electrotransfection efficiency of Huh-7 cells. Compared to chemical based reagents, electroporation showed the superior levels of transfection in both cell lines. The presented protocols should satisfy most of the experimental applications requiring high transfection efficiencies of these two cell lines. PMID:23285746

Hashemi, A; Roohvand, F; Ghahremani, M H; Aghasadeghi, M R; Vahabpour, R; Motevau, F; Memarnejadian, A

2012-01-01

33

Structure-Function Relationship of the Ion Channel Formed by Diphtheria Toxin in Vero Cell Membranes  

Microsoft Academic Search

.   Diphtheria toxin (DT) forms cation selective channels at low pH in cell membranes and planar bilayers. The channels formed\\u000a by wild-type full length toxin (DT-AB), wild-type fragment B (DT-B) and mutants of DT-B were studied in the plasma membrane\\u000a of Vero cells using the patch-clamp technique. The mutations concerned certain negatively charged amino acids within the channel-forming\\u000a transmembrane domain

M. Lanzrein; P. Ø. Falnes; O. Sand; S. Olsnes

1997-01-01

34

[Fluorescent derivatives of diphtheria toxin subunit B and their interaction with Vero cells].  

PubMed

Diphtheria toxin's B subunit provides toxin interaction with its receptor on the cell surface and translocation of toxin's A subunit from endosome to cytozole of sensitive cells. Functional analogues of B subunit with fluorescent label are considered as perspective tools for studying the above mentioned processes. The aim of the work was to obtain fluorescent B subunit analogues and to detect the specificity of their interaction with Vero line cells. B subunit fluorescent analogues were obtained in two different ways. The first one was B subunit chemical conjugation with fluorescein isothiocyanate and the second one was genetic fusion of recombinant B subunit chain with enhanced green fluorescent protein chain. Specific interaction of B subunit fluorescent derivatives with Vero cells was studied by flow cytometry and confocal microscopy. Using competitive analysis it was shown that B subunit fluorescent analogues possessed different affinity for cells. The affinity of EGFP-SbB was higher than FITC-SbB. Our results indicate the possibility to use the fluorescent derivatives of B subunit as tools for identification of diphtheria toxin's receptor (HB-EGF) expression on the cell surface as well as for studying the interaction and penetration of diphtheria toxin to the cell. PMID:19877418

Kaberniuk, A A; Labyntsev, A Iu; Kolybo, D V; Oli?nyk, O S; Redchuk, T A; Korotkevych, N V; Horchev, V F; Karakhim, S O; Komisarenko, S V

2009-01-01

35

Prevention of lipid peroxidation induced by ochratoxin A in Vero cells in culture by several agents.  

PubMed

Ochratoxin A (OTA) is a mycotoxin produced by Aspergillus ochraceus as well as other moulds. This mycotoxin contaminates animal feed and food and is nephrotoxic for all animal species studied so far. OTA is immunosuppressive, genotoxic, teratogenic and carcinogenic. It is a structural analogue of phenylalanine and contains a chlorinated dihydroisocoumarinic moiety. Ochratoxin A inhibits protein synthesis by competition with phenylalanine in the phenylalanine-tRNA aminoacylation reaction. Recently lipid peroxidation induced by OTA has been reported, indicating that the lesions induced by this toxin could also be related to oxidative damage. An attempt to prevent its toxic effect, mainly the lipid peroxidation, has been made using aspartame (L-aspartyl-L-phenylalanine methyl ester) a structural analogue of both OTA and phenylalanine, piroxicam, a non steroidal anti-inflammatory drug and superoxide dismutase+catalase (endogenous oxygen radical scavengers). Lipid peroxidation was assayed in monkey kidney cells (Vero cells) treated by increasing concentrations of OTA (5-50 microM). After 24 h incubation OTA induced lipid peroxidation in Vero cells in a concentration dependent manner, as measured by malonaldehyde (MDA) production. The MDA production, in Vero cells, was significantly increased by 50.5% from 694.1 +/- 21.0 to 1041.5 +/- 23.5 pmol/mg of protein. In the presence of superoxide dismutase (SOD)+catalase (25 micrograms/ml each) the MDA production induced by OTA was significantly decreased. At 50 microM of OTA concentration (optimal production of MDA) the MDA production decreased from 1041.5 +/- 23.5 to 827.5 +/- 21.3 pmol/mg of protein. SOD and catalase, when applied prior to the toxin, seemed to prevent lipid peroxidation more efficiently than piroxicam (at a ten-fold higher concentration than OTA) and aspartame (at equimolar concentration). These molecules also partially prevented the OTA-induced leakage of MDA in the culture medium. PMID:9158693

Baudrimont, I; Ahouandjivo, R; Creppy, E E

1997-04-18

36

Vero cells expressing porcine circovirus type 2-capsid protein and their diagnostic application.  

PubMed

Porcine circovirus type 2 (PCV2) is the causative agent of postweaning multisystemic wasting syndrome (PMWS) in swine. Although the incidences of PCV2-related diseases are ubiquitous throughout the world, the serological tools are rather limited, mainly because the virus does not induce any cytopathic effects in cells. The purpose of this study was to develop a rapid, sensitive and easy quantitative immunofluorescence assay (QIFA) using the recombinant PCV2 nucleocapsid protein (NCP) for the detection of PCV2-specific antibodies in pig sera. The recombinant PCV2 NCP was expressed in Vero cells by a lentivirus system. The performance of QIFA using these Vero cells as a diagnostic antigen was compared with currently available C-ELISA and I-ELISA; the relative sensitivity turned out to range from 92.5% up to 99.3%. The relative specificity was 93.3% when compared to C-ELISA as the gold standard. The serological experiment also indicated the inverse relationship between QIFA and the viral load in serum, semen, feces samples from 7 PCV2-positive boars. In addition, the PCV2 sequence detected from bone marrow cells shows 99% of sequence identity with PCV2 genome, confirming the infectivity of PCV2. PMID:23954842

Kim, Yeon-Hee; Kweon, Chang-Hee; Kang, Seung-Won; Oh, Yoon I; Song, Jae Young; Lee, Kyoung Ki; Park, Se Chang

2013-12-01

37

Immunological equivalence between mouse brain-derived and Vero cell-derived Japanese encephalitis vaccines.  

PubMed

The persistent spread via animal reservoirs urges expanding vaccination programs against pathogens like the Japanese encephalitis virus, JEV. The JEV is spreads to new areas by domestic as well as by wild animals. Although there is a safe and efficient vaccine on the market, this is derived from infected mouse brains, why today's situation requires overcoming the potential risk caused by using animal tissues. To meet this demand we have developed a Vero cell-derived JEV vaccine, using the same virus strain as in the established one. A phase III clinical study of the new vaccine has recently been completed with positive outcome. Like the established mouse brain-derived vaccine, the Vero cell-derived one is a formalin inactivated whole virus vaccine. We here demonstrate the very good agreement in immunological tests between the two antigens. The study includes analyses with two neutralizing monoclonal antibodies that blocks cell entry at a late stage in infection, assumedly interfering with fusion-related refolding in the virus fusion protein. It is obvious that the formalin inactivation treatment, with both virus preparations, retains these essential vaccine epitopes. PMID:16815584

Abe, Motoharu; Shiosaki, Kouichi; Hammar, Lena; Sonoda, Kengo; Xing, Li; Kuzuhara, Syoji; Kino, Yoichiro; Holland Cheng, R

2006-11-01

38

A purified inactivated Japanese encephalitis virus vaccine made in Vero cells.  

PubMed

A second generation, purified, inactivated vaccine (PIV) against Japanese encephalitis (JE) virus was produced and tested in mice where it was found to be highly immunogenic and protective. The JE-PIV was made from an attenuated strain of JE virus propagated in certified Vero cells, purified, and inactivated with formalin. Its manufacture followed current GMP guidelines for the production of biologicals. The manufacturing process was efficient in generating a high yield of virus, essentially free of contaminating host cell proteins and nucleic acids. The PIV was formulated with aluminum hydroxide and administered to mice by subcutaneous inoculation. Vaccinated animals developed high-titered JE virus neutralizing antibodies in a dose dependent fashion after two injections. The vaccine protected mice against morbidity and mortality after challenge with live, virulent, JE virus. Compared with the existing licensed mouse brain-derived vaccine, JE-Vax, the Vero cell-derived JE-PIV was more immunogenic and as effective as preventing encephalitis in mice. The JE-PIV is currently being tested for safety and immunogenicity in volunteers. PMID:11483284

Srivastava, A K; Putnak, J R; Lee, S H; Hong, S P; Moon, S B; Barvir, D A; Zhao, B; Olson, R A; Kim, S O; Yoo, W D; Towle, A C; Vaughn, D W; Innis, B L; Eckels, K H

2001-08-14

39

Non-clinical and phase I clinical trials of a Vero cell-derived inactivated Japanese encephalitis vaccine.  

PubMed

The safety and effectiveness of a Vero cell-derived inactivated Japanese encephalitis (JE) vaccine were compared with those of a current JE vaccine in non-clinical studies and a phase I clinical trial. The single-dose toxicity study showed no toxicity of either the current JE vaccine or the investigational Vero cell-derived JE vaccine. In a local irritation study, the degree of irritation caused by both vaccines was determined to be the same as that induced by normal saline. To investigate genotoxicity, a chromosomal aberration test was conducted and the results were negative. Both JE vaccines were administered to a group of 30 subjects who were seronegative (neutralizing antibody titer <10(1)) for JEV virus (Beijing-1 Strain). Each subject was subcutaneously inoculated twice at an interval of 1-4 weeks, followed by an additional booster inoculation 4-8 weeks later, and clinical reactions and serological responses were subsequently investigated. Adverse drug reactions of local reaction, headache and malaise were mild, occurring at a rate of 6.7 and 20.0% after administration of the Vero cell-derived JE vaccine and the current JE vaccine, respectively. The seroconversion rate after three doses of both JE vaccines was 100%, while the geometric mean titer for the Vero cell-derived and current JE vaccines was 10(2.35) and 10(2.03), respectively. These results suggest that the safety and effectiveness of the Vero cell-derived inactivated JE vaccine are equal to those of the currently available conventional vaccine in humans, and that the Vero cell-derived vaccine could be a useful second-generation JE vaccine. PMID:14575762

Kuzuhara, Syoji; Nakamura, Hideki; Hayashida, Kenshi; Obata, Junko; Abe, Motoharu; Sonoda, Kengo; Nishiyama, Kiyoto; Sugawara, Keishin; Takeda, Kengo; Honda, Tomitaka; Matsui, Hajime; Shigaki, Takamichi; Kino, Yoichiro; Mizokami, Hiroshi; Tanaka, Masahiko; Mizuno, Kyosuke; Ueda, Kohji

2003-11-01

40

Adaptation of High-Growth Influenza H5N1 Vaccine Virus in Vero Cells: Implications for Pandemic Preparedness  

PubMed Central

Current egg-based influenza vaccine production technology can't promptly meet the global demand during an influenza pandemic as shown in the 2009 H1N1 pandemic. Moreover, its manufacturing capacity would be vulnerable during pandemics caused by highly pathogenic avian influenza viruses. Therefore, vaccine production using mammalian cell technology is becoming attractive. Current influenza H5N1 vaccine strain (NIBRG-14), a reassortant virus between A/Vietnam/1194/2004 (H5N1) virus and egg-adapted high-growth A/PR/8/1934 virus, could grow efficiently in eggs and MDCK cells but not Vero cells which is the most popular cell line for manufacturing human vaccines. After serial passages and plaque purifications of the NIBRG-14 vaccine virus in Vero cells, one high-growth virus strain (Vero-15) was generated and can grow over 108 TCID50/ml. In conclusion, one high-growth H5N1 vaccine virus was generated in Vero cells, which can be used to manufacture influenza H5N1 vaccines and prepare reassortant vaccine viruses for other influenza A subtypes.

Huang, Mei-Liang; Yeh, Wei-Zhou; Weng, Tsai-Chuan; Chen, Yu-Shuan; Chong, Pele; Lee, Min-Shi

2011-01-01

41

Development of a Vero cell-derived influenza whole virus vaccine.  

PubMed

Influenza vaccine production is dependent on the availability of embryonated hen eggs for virus growth. This is an extremely cumbersome system with many disadvantages with respect to selection of virus variants and the presence of adventitious viruses. We have developed an alternative cell culture system which allows rapid production of large volumes of vaccine. The WHO-approved Vero cell line was used in serum-free culture to grow many influenza strains to high titre. This system could be scaled-up to allow vaccine production with a 1200 litre fermenter. A purification scheme was developed which resulted in a high purity whole virus vaccine. This was demonstrated to be at least as immunogenic as a conventional egg-derived preparation. PMID:10494963

Kistner, O; Barrett, P N; Mundt, W; Reiter, M; Schober-Bendixen, S; Eder, G; Dorner, F

1999-01-01

42

Characterization of Vero cell growth and death in bioreactor with serum-containing and serum-free media  

Microsoft Academic Search

The density of viable cells in a culture results from a balance between cell proliferation and cell death. The aim of this\\u000a study was to characterize and compare these two phenomena in Vero cell cultures in one serum containing medium (ScA) and one\\u000a serum free medium (SfB) in bioreactors. Cell growth was evaluated by cell counting(after crystal violet staining) and

Sébastien Quesney; Jacqueline Marvel; Annie Marc; Catherine Gerdil; Bernard Meignier

2001-01-01

43

Genetic stability of oral polio vaccine prepared on primary monkey kidney cells or Vero cells--effects of passage in cell culture and the human gastrointestinal tract.  

PubMed

The genetic stability of the three Sabin oral poliovaccine (OPV) strains produced on either primary monkey kidney (VK) or Vero cell substrates was compared in vivo and in vitro by measuring the rate at which the bases most strongly associated with attenuation and reversion to neurovirulence (positions 480, 481, and 472 in the 5' non-coding region of Sabin 1, 2 and 3 respectively, and 2034 in VP3 of Sabin 3) reverted during passage of the vaccine strains in the gastrointestinal tract of primary vaccinees and in cell culture. For the in vivo study, the poliovirus excretion patterns of 21 infants receiving OPV produced on either VK or Vero cells were followed for 21 days. No significant differences in either the frequency of excretion or the rate of reversion were observed between the two vaccine groups. The rate of accumulation of revertants during passage in vitro was compared for the three Sabin strains passaged 10 times in either VK or Vero cells. For types 1 and 3, revertants accumulated faster upon passage through VK cells compared with passage through Vero cells. Type 2 appeared to be stable as no revertants were detected in either cell type. Results of this study suggest that the use of Vero as opposed to VK cells as substrate for the manufacture of OPV does not negatively influence the genetic stability of the three Sabin OPV strains in vivo or in vitro. PMID:9796061

Chezzi, C; Dommann, C J; Blackburn, N K; Maselesele, E; McAnerney, J; Schoub, B D

1998-12-01

44

Immunogenicity of single-dose Vero cell-derived Japanese encephalitis vaccine in Japanese adults.  

PubMed

In Japan, intensive immunization against Japanese encephalitis (JE) was performed from 1967 to 1976, and regular JE immunization was performed thereafter. However, for Japanese adults facing JE risk, dates of vaccination with new inactivated Vero cell-derived JE vaccine are unavailable. This study investigated how a single dose of Vero cell-derived JE vaccine affects Japanese adults. Neutralizing antibodies were measured pre- and post-JE vaccination in 79 participants (age 40.7 ± 9.4 years), enrolled between October 2009 and March 2011, whose JE-vaccination data were gathered from vaccination records and history taking. Before vaccination, the participants' seroprotection rate (SPR) was 51.9%, whereas SPR after vaccination was 93.7%. The seroconversion rate (SCR), which measures seronegative cases that turn seropositive after vaccination, was 86.8%. The geometric mean titer (GMT) was 14.7 before vaccination and 70.1 after vaccination. Age was a significant difference between seroprotected (42.8 years) and non-seroprotected (38.7 years) groups before vaccination. Then the difference of age, SCR, pre-vaccination GMT, post-vaccination GMT and sex ratio were also significant in participants aged 25-39 years and ?40 years, who represent generations born when Japan's JE-vaccination policy changed. SCR was 100% in participants aged 25-39 years with a vaccination recorded 55.6% in participants aged 25-39 without a vaccination record, and 96.0% in participants aged ?40 years. Thus, more participants aged 25-39 years were seroprotected before vaccination, but SCR was higher in those aged ?40 years. Most Japanese adults can be protected after one-dose vaccination, but this may be insufficient for people aged 25-39 years without recorded JE vaccination. PMID:24485326

Takeshita, Nozomi; Lim, Chang-Kweng; Mizuno, Yasutaka; Shimbo, Takuro; Kotaki, Akira; Ujiie, Mugen; Hayakawa, Kayoko; Kato, Yasuyuki; Kanagawa, Shuzo; Kaku, Mitsuo; Takasaki, Tomohiko

2014-04-01

45

The effect of serial passage on Sabin oral poliomyelitis vaccine virus in secondary monkey kidney versus Vero cells as measured by the monkey neurovirulence test.  

PubMed

The three Sabin strains of poliomyelitis seed virus were serially passaged in either secondary monkey kidney or Vero cell cultures and the tenth passage of each virus harvest compared to non-passaged Sabin reference virus of the same type using the monkey neurovirulence test. All three types were further attenuated by passage in Vero cells, whereas only type 2 became further attenuated after passage in secondary monkey kidney cells. After passage in Vero cells, type 3 poliomyelitis virus became more heat stable, as measured by its replicative capacity at 40 degrees C. PMID:7731028

Dommann, C J

1994-09-01

46

Canine distemper virus induces apoptosis through caspase-3 and -8 activation in vero cells.  

PubMed

We investigated the signal-transduction pathway of canine distemper virus-Onderstepoort (CDV-Ond) vaccine strain-mediated apoptosis in Vero cells. Canine distemper virus-Onderstepoort at a multiplicity of infection (MOI) of 0.1 induced DNA fragmentation 48 h after infection. Immunofluorescence staining revealed that 57% +/- 4% of the CDV-N-protein-positive cells were terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)-positive, and all TUNEL-positive cells were CDV-N-protein-positive, indicating that CDV-Ond induced apoptosis only in the infected cells. We also found that CDV-Ond infection induced activation of caspase-3 and caspase-8. In the semi-quantitative reverse transcription-polymerase chain reaction assay for apoptosis-related genes, the expression of mRNA of the death receptor, Fas, was also increased in CDV-Ond-infected cells. In contrast, the expressions of Bcl-2 and Bax, regulators for intrinsic apoptotic signaling through the mitochondria, did not change. These results suggest that CDV-Ond induced apoptosis by activating caspase-3, possibly through caspase-8 signaling rather than through p53/Bax-mediated, mitochondrial signaling in the infected cells. PMID:16907958

Kajita, M; Katayama, H; Murata, T; Kai, C; Hori, M; Ozaki, H

2006-08-01

47

Label free detection of pseudorabies virus infection in Vero cells using laser force analysis.  

PubMed

The rapid and robust identification of viral infections has broad implications for a number of fields, including medicine, biotechnology and biodefense. Most detection systems rely on specific molecules, such as nucleic acids or proteins, to identify the target(s) of interest. These molecules afford great specificity, but are often expensive, labor-intensive, labile and limited in scope. Label free detection methods seek to overcome these limitations by instead using detection methods that rely on intrinsic properties as a basis for identifying and separating species of interest and thus do not rely on specific prior knowledge of the target. Optical chromatography, one such technique, uses the balance between optical and fluidic drag forces within a microfluidic channel to determine the optical force on cells or particles. Here we present the application of individual optical force measurements as a means of investigating pseudorabies virus infection in Vero cells. Optical force differences are seen between cells from uninfected and infected populations at a multiplicity of infection as low as 0.001 and as soon as 2 hours post infection, demonstrating the potential of this technique as a means of detecting viral infection. Through the application of a pattern recognition neural network, individual cell size data are combined with optical force as a means of classifying cell populations. Potential applications include the early detection of bloodborne pathogens for the prevention of sepsis and other diseases as well as the detection of biological threat agents. PMID:24492491

Hebert, Colin G; Hart, Sean J; Terray, Alex

2014-03-21

48

Role of kinesin-1 and cytoplasmic dynein in endoplasmic reticulum movement in VERO cells  

PubMed Central

Summary Generating the extended endoplasmic reticulum (ER) network depends on microtubules, which act as tracks for motor-driven ER tubule movement, generate the force to extend ER tubules by means of attachment to growing microtubule plus-ends and provide static attachment points. We have analysed ER dynamics in living VERO cells and find that most ER tubule extension is driven by microtubule motors. Surprisingly, we observe that ?50% of rapid ER tubule movements occur in the direction of the centre of the cell, driven by cytoplasmic dynein. Inhibition of this movement leads to an accumulation of lamellar ER in the cell periphery. By expressing dominant-negative kinesin-1 constructs, we show that kinesin-1 drives ER tubule extension towards the cell periphery and that this motility is dependent on the KLC1B kinesin light chain splice form but not on KLC1D. Inhibition of kinesin-1 promotes a shift from tubular to lamellar morphology and slows down the recovery of the ER network after microtubule depolymerisation and regrowth. These observations reconcile previous conflicting studies of kinesin-1 function in ER motility in vivo. Furthermore, our data reveal that cytoplasmic dynein plays a role in ER motility in a mammalian cultured cell, demonstrating that ER motility is more complex than previously thought.

Wozniak, Marcin J.; Bola, Becky; Brownhill, Kim; Yang, Yen-Ching; Levakova, Vesselina; Allan, Victoria J.

2009-01-01

49

Photoirradiation study of gold nanospheres and rods in Vero and Hela cell lines  

NASA Astrophysics Data System (ADS)

Photoirradiation effect of gold nanospheres in conjucation with green light and rods in conjugation with red light corresponds to their absorption wavelength range found to be appreciable. In this present work concentration of nanomaterial and light dose were optimized. Gold nanospheres were synthesized by reduction technique using Sodium Borohydrate as reducing agent and Trisodium Citrate as capping agent. Au nanorods having 680-900nm absorption were synthesized using reduction techniques with CTAB and BDAC polymers. From UV-Vis absorption and Transmission Electron Microscopy the size of nanoparticles were confirmed. 30nm Gold nanospheres and green light source of 530nm wavelength with power 30mW were applied to Vero and Hela cell lines shows higher toxicity for Hela cells. Nanorods were applied and irradiated with 680nm wavelength light source with light intensity 45mW. Post irradiation effect for 24hrs, 48hrs confirms cell proliferation in normal rate in viable cells. The morphological changes in irradiated spot leads to apoptotoic cell death was confirmed with microscopic imaging. The LD50 value was also calculated.

Gananathan, Poorani; Aruna, Prakasarao; Ganesan, Singaravelu; Elanchezhiyan, Manickan

2014-03-01

50

Lipophilic organic pollutants induce changes in phospholipid and membrane protein composition leading to Vero cell morphological change.  

PubMed

Membrane damage related to morphological change in Vero cells is a sensitive index of the composite biotoxicity of trace lipophilic chemicals. However, judging whether the morphological change in Vero cells happens and its ratio are difficult because it is not a quantitative characteristic. To find biomarkers of cell morphological change for quantitatively representing the ratio of morphological changed cell, the mechanism of cell membrane damage driven by typical lipophilic chemicals, such as trichlorophenol (TCP) and perfluorooctanesulphonate (PFOS), was explored. The ratio of morphologically changed cells generally increased with increased TCP or PFOS concentrations, and the level of four major components of phospholipids varied with concentrations of TCP or PFOS, but only the ratio of phosphatidylcholine (PC)/phosphatidylethanolamine (PE) decreased regularly as TCP or PFOS concentrations increased. Analysis of membrane proteins showed that the level of vimentin in normal cell membranes is high, while it decreases or vanishes after TCP exposure. These variations in phospholipid and membrane protein components may result in membrane leakage and variation in rigid structure, which leads to changes in cell morphology. Therefore, the ratio of PC/PE and amount of vimentin may be potential biomarkers for representing the ratio of morphological changed Vero cell introduced by trace lipophilic compounds, thus their composite bio-toxicity. PMID:25065828

Liao, Ting T; Wang, Lei; Jia, Ru W; Fu, Xiao H; Chua, Hong

2014-10-01

51

Development of an animal-component free medium for vero cells culture.  

PubMed

This work describes the development of an animal-component free medium (IPT-AFM) that allows an optimal growth of Vero cells, an adherent cell line used for the production of viral vaccines. Statistical experimental design was applied to identify crucial nutrients that affect cell growth. Using Medium 199 or MEM as a basal medium, a serum-free medium (SFM) referred as IPT-SFM that only enclosed transferrin as a component of animal origin was developed at first. Then, the composition of IPT-SFM was further improved to obtain an animal-component free medium named IPT-AFM. IPT-AFM contains M199 as a basal medium, plant hydrolysates, epidermal growth factor, ethanolamine, ferric citrate, and vitamin C. Among various plant hydrolysates, specific combinations of soy (Hypep 1510) and wheat gluten (Hypeps 4601 and 4605) hydrolysates, were identified to promote cell growth; whereas individual Hypeps had a minor positive effect on cell growth. Nevertheless, the removal of serum did influence cell attachment. Coating tissue-culture flasks with teleostean, a product extracted from cold water fish skin, had not only enhanced cell attachment but also improved cell growth performance in static cultures. Different non-animal proteases were also assessed as an alternative to trypsin. TrypLE Select, a recombinant trypsin, gave the best cell growth performances. Kinetics of cell growth in IPT-AFM were investigated in T-flasks, cell growth was comparable with that obtained in MEM+10% fetal calf serum (FCS). A mean cell division number equal to 2.26 +/- 0.18 and a specific growth rate micro 0.019 +/- 0.003 h(-1) were achieved in IPT-AFM. PMID:19768803

Rourou, Samia; van der Ark, Arno; van der Velden, Tiny; Kallel, Héla

2009-01-01

52

Comparison of viral glycosylation using lectin blotting with Vero cell-derived and mouse brain-derived Japanese encephalitis vaccines.  

PubMed

We produced a Vero cell-derived inactivated Japanese encephalitis vaccine using a serum-free medium, as a substitute for the conventional mouse brain-derived Japanese encephalitis vaccine. The immunogenicity of this cell-derived vaccine was higher than that of the conventional mouse brain-derived vaccine. The results of a clinical study in humans also demonstrated higher immunogenicity of this cell-derived vaccine. No gene mutation was found in the viral structural proteins derived from Vero cells and mouse brain. So, we conducted a lectin blot analysis, assuming differing glycosylation as a cause of the higher immunogenicity in humans. The results demonstrated that vaccine reactivity varied with lectins, particularly with WGA, DBA, MAM, SSA, SBA, and GS-II. Thus, glycosylation differed with the vaccines, suggesting a possible cause of the differing immunogenicity between mice and humans. PMID:21195800

Toriniwa, Hiroko; Komiya, Tomoyoshi

2011-02-24

53

Identification of diphtheria toxin receptor and a nonproteinous diphtheria toxin-binding molecule in Vero cell membrane  

PubMed Central

Two substances possessing the ability to bind to diphtheria toxin (DT) were found to be present in a membrane fraction from DT-sensitive Vero cells. One of these substances was found on the basis of its ability to bind DT and inhibit its cytotoxic effect. This inhibitory substance competitively inhibited the binding of DT to Vero cells. However this inhibitor could not bind to CRM197, the product of a missense mutation in the DT gene, and did not inhibit the binding of CRM197 to Vero cells. Moreover, similar levels of the inhibitory activity were observed in membrane fractions from DT-insensitive mouse cells, suggesting the inhibitor is not the DT receptor which is specifically present in DT-sensitive cells. The second DT-binding substance was found in the same Vero cell membrane preparation by assaying the binding of 125I-labeled CRM197. Such DT-binding activity could not be observed in membrane preparation from mouse L cells. From competition studies using labeled DT and CRM proteins, we conclude that this binding activity is due to the surface receptor for DT. Treatment of these substances with several enzymes revealed that the inhibitor was sensitive to certain RNases but resistant to proteases, whereas the DT receptor was resistant to RNase but sensitive to proteases. The receptor was solubilized and partially purified by chromatography on CM- Sepharose column. Immunoprecipitation and Western blotting analysis of the partially purified receptor revealed that a 14.5-kD protein is the DT receptor, or at least a component of it.

1988-01-01

54

Long-term stability of Vero cell-derived inactivated Japanese encephalitis vaccine prepared using serum-free medium.  

PubMed

We established a method of producing a Vero cell-derived Japanese encephalitis vaccine using serum-free medium, and tested its stability using various stabilizers during the inactivation process and storage at 4 degrees C and 28 degrees C. Similar to previously reported results of cell culture in serum-containing medium, Vero cells were cultured in a serum-free medium multiplied well, and the viral yield was successfully increased to about 10(9)PFU/ml. Following formalin-inactivation and purification via ethanol precipitation and sucrose density ultracentrifugation of the virus solution, the vaccine had the same quality as, and higher immunogenicity, the mouse brain-derived vaccine in current use. Testing of several stabilizers showed that the addition of 0.5% glycine during the virus inactivation process facilitated the maintenance of immunogenicity for a long period of time. Furthermore, the addition of 0.5% glycine and 1.0% sorbitol as vaccine stabilizers after purification led to the maintenance of immunogenicity for 1 year, not dependent on the storage temperature (4 degrees C or 28 degrees C). These results indicate that, in contrast to the current mouse brain-derived vaccine, the Vero cell-derived vaccine can be prepared using serum-free medium containing no animal-derived components, and that the vaccine can be stored at room temperature by adding stabilizers, suggesting the possibility of producing room temperature-stable vaccines. PMID:18534722

Toriniwa, Hiroko; Komiya, Tomoyoshi

2008-07-01

55

Safety profile of the Vero cell-derived Japanese encephalitis virus (JEV) vaccine IXIARO(®).  

PubMed

Japanese encephalitis (JE) is the most common cause for viral encephalitis in Asia and can be effectively prevented by vaccination. IXIARO(®) is a Vero cell-derived, inactivated JE virus vaccine which has been licensed and distributed in the US, Europe, Canada, Hongkong, Israel, and distributed in Australia under the trade name JESPECT(®). This paper reviews the safety profile of IXIARO(®) in the first 12months after licensure and discusses the observed profile in the context of clinical trial results for IXIARO(®) and post-marketing safety data for JE-VAX(®). The clinical safety profile is derived from a pooled analysis including safety data from 10 phase III trials in 4043 subjects who received at least one IXIARO(®) vaccination and were followed-up for up to 3years after the primary immunization. Local and systemic tolerability of IXIARO(®) was similar to an earlier safety analysis at the time of licensure of the vaccine. In post-marketing AE reports, the system organ classes affected following vaccination with IXIARO(®) were similar to the previously observed clinical trial profile. No serious allergic reactions were observed in the 12-month post-marketing period. This comprehensive safety review confirms the good safety profile of IXIARO(®) in clinical and post-marketing use. PMID:21907747

Schuller, Elisabeth; Klingler, Anton; Dubischar-Kastner, Katrin; Dewasthaly, Shailesh; Müller, Zsuzsanna

2011-11-01

56

In-vitro maturation of round spermatids using co-culture on Vero cells.  

PubMed

In an attempt to determine whether co-culture could promote sperm maturation, three patients with non-obstructive azoospermia, two with maturation arrest at the level of primary spermatocytes and one patient with <1% tubules showing complete spermatogenesis, and one patient with total globozoospermia, gave consent to experimentally co-culture round spermatids retrieved from the testicle on Vero cell monolayers. In all azoospermic patients elongating spermatids could be obtained from round spermatids. In one case of maturation arrest, of 37 round spermatids co-cultured for up to 5 days, 30% developed flagella, 46% matured to elongating and 19% to elongated spermatids, with one mature spermatozoon also obtained (3%). In the same patient, primary cultures of three round spermatids with flagella enabled development of one further mature spermatozoon. In the case with total globozoospermia, of six round spermatids co-cultured for up to 5 days, one mature spermatozoon was obtained, with a flagellum and normal head morphology. These preliminary findings suggest that it may be possible to overcome the round spermatid block, and even the triggering of morphological abnormalities arising at the spermiogenic level, by in-vitro maturation under special environmental conditions. PMID:10325279

Cremades, N; Bernabeu, R; Barros, A; Sousa, M

1999-05-01

57

Structural and functional analysis of virus factories purified from Rabbit vesivirus-infected Vero cells.  

PubMed

Rabbit vesivirus infection induces membrane modifications and accumulation of vesicular structures in the cytoplasm of infected Vero cells. Crude RaV replication complexes (RCs) have been purified and their structural and functional properties have been characterized. We show that calnexin, an ER-resident protein, RaV non-structural proteins 2AB-, 2C-, 3A-, 3B- and 3CD-like as well as viral RNAs co-localize within membranous structures which are able to replicate the endogenous RNA templates. The purified virus factories protected their viral RNA contents from microccocal nuclease degradation and were inaccessible to exogenously added synthetic transcripts. In addition, we have shown that RCs can be used to investigate uridylylation of native endogenous VPg. In contrast to the observation that the virus factories were inaccessible to RNAs, RCs were accessible to added recombinant VPg which was subsequently nucleotidylylated. Nevertheless no elongation of an RNA chain attached to native or recombinant VPg could be demonstrated. PMID:18625274

Casais, Rosa; Molleda, Lorenzo González; Machín, Angeles; del Barrio, Gloria; Manso, Alberto García; Dalton, Kevin P; Coto, Ana; Alonso, José Manuel Martín; Prieto, Miguel; Parra, Francisco

2008-10-01

58

Comparison of use of Vero cell line and suspension culture of murine macrophage to attenuation of virulence of Neospora caninum.  

PubMed

In this study the tachyzoite yields of Neospora caninum were compared in two cell lines: Vero (African Green Monkey Kidney) and suspension culture of murine macrophage (J774) cell lines. Then, N. caninum were continuously passaged in these cell lines for 3 months and the effect of host cells on virulence of tachyzoites was assessed by broiler chicken embryonated eggs. Inoculation was performed in the chorioallantoic (CA) liquid of the embryonated eggs with different dilutions (0.5 × 10(4), 1.0 × 10(4), 1.5 × 10(4)) of tachtzoites isolated from these cell cultures. The mortality pattern and pathological changes of the dead embryos and hatched chickens were noted. Tissue samples of brain, liver and heart were examined by histopathological and detection of DNA of parasite by polymerase chain reaction (PCR). Also, consecutive sections of the tissues examined histologically were used for immunohistochemical (IHC) examination. Embryos inoculated with tachyzoites derived from Vero cell line (group V) showed a higher mortality rate (100%) than the embryos that received tachyzoites derived from J774 cell line (group J) (10% mortality rate). The results of this study indicated that the culture of N. caninum in J774 cell led to a marked increase in the number of tachyzoite yields and rapid attenuation in comparison to Vero, so the results were confirmed by IHC and PCR. This study is the first report of the significant effect of host cell on the attenuation of virulence of N. caninum tachyzoites. These findings could potentially provide a practical approach in the mass production of N. caninum tachyzoites, and also in producing live attenuated vaccine. PMID:23684321

Khordadmehr, Monireh; Namavari, Mehdi; Khodakaram-Tafti, Azizollah; Mansourian, Maryam; Rahimian, Abdollah; Daneshbod, Yahya

2013-10-01

59

Phorbol Esters Isolated from Jatropha Meal Induced Apoptosis-Mediated Inhibition in Proliferation of Chang and Vero Cell Lines  

PubMed Central

The direct feeding of Jatropha meal containing phorbol esters (PEs) indicated mild to severe toxicity symptoms in various organs of different animals. However, limited information is available on cellular and molecular mechanism of toxicity caused by PEs present in Jatropha meal. Thus, the present study was conducted to determine the cytotoxic and mode of action of PEs isolated from Jatropha meal using human hepatocyte (Chang) and African green monkey kidney (Vero) cell lines. The results showed that isolated PEs inhibited cell proliferation in a dose-dependent manner in both cell lines with the CC50 of 125.9 and 110.3 ?g/mL, respectively. These values were compatible to that of phorbol 12-myristate 13-acetate (PMA) values as positive control i.e., 124.5 and 106.3 ?g/mL respectively. Microscopic examination, flow cytometry and DNA fragmentation results confirmed cell death due to apoptosis upon treatment with PEs and PMA at CC50 concentration for 24 h in both cell lines. The Western blot analysis revealed the overexpression of PKC-? and activation of caspase-3 proteins which could be involved in the mechanism of action of PEs and PMA. Consequently, the PEs isolated form Jatropha meal caused toxicity and induced apoptosis-mediated proliferation inhibition toward Chang and Vero cell lines involving over-expression of PKC-? and caspase-3 as their mode of actions.

Oskoueian, Ehsan; Abdullah, Norhani; Ahmad, Syahida

2012-01-01

60

Evaluation of the serum-free medium MDSS2 for the production of poliovirus on vero cells in bioreactors  

Microsoft Academic Search

The serum-free medium MDSS2 (Merten et al., 1994), was used for cultivating Vero cells as well as for producing poliovirus\\u000a (Sabin type 1) in static and in perfused micro-carrier cultures. At slightly different growth rates of 0.0120\\/h and 0.0106\\/h,\\u000a respectively, static cultures in serum-containing (SCM) and serum-free (SFM) medium produced titers of 106.75 and 106.67 TCID50 per 50 µl; signifying

O.-W. Merten; R. Wu; E. Couvé; R. Crainic

1997-01-01

61

Systematically experimental investigation on carcinogenesis or tumorigenicity of VERO cell lines of different karyotypes in nude mice in vivo used for viral vaccine manufacture.  

PubMed

Many cell lines used for vaccine production have a potentially strong tumorigenic character. Some of those routinely used need to be checked at different passage numbers for this characteristic. Using HeLa cell cultures as positive controls, and primary canine kidney cell (CKC) or feline kidney cell (FKC) cultures purified in vitro on passage three as negative controls, the tumorigenicity of VERO cell sublines was tested in 219 nude mice. The master cell stocks (MCS) and working cell banks (WCB) of eight strains of VERO African green monkey kidney cell (AGMKC) line used for canine, feline and mink vaccine preparation were established in China. The hypo-tetra-ploid JA or hyper-diploid KA strain of VERO line was highly tumorigenic. These data showed a variable chromosome karyotype of VERO line, and contraindicated the use of JA or KA strain of VERO line for the preparation of attenuated viral vaccines. JA or KA strain of VERO line could be a substitute for HeLa line as a positive-control malignant tumor (MT) cell model. The non-carcinogenic YB, JC, M and JB strains of VERO line were therefore selected for the preparation of modified live rabies viral vaccine in place of BHK-21. The cell sub-lines are comparatively stable in terms of their heritable characters, and show little significant changes between passages. In summary, we have found that: 1) the tumorigenicity of cell line is different among different-karyotypic cells; 2) it is the genetic characteristics of chromosomes of cell lines that determines their tumorigenicity, but with species-specific carcinogenicity; 3) the chromosome number variation of cell lines has positive relationship with their carcinogenesis; 4) highly variable strains of tumor cell line can be selected quickly and successfully in nude mice by alternate cultivation in vitro and in vivo. Malignant rhabdoid tumor (MRT) was evolved in nude mice inoculated with violently variable HeLa or VERO cells. The importance of assessing the tumorigenicity in cell sublines used for vaccine production is emphasised. PMID:15473315

Zhang, De-Li; Ji, Liang; Li, Liu-Jin; Huang, Gao-Sheng

2004-07-01

62

Autophagic Cell Death Is Induced by Acetone and Ethyl Acetate Extracts from Eupatorium odoratum In Vitro: Effects on MCF-7 and Vero Cell Lines  

PubMed Central

Eupatorium odoratum (EO) contains many biologically active compounds, the anticancer effects of which are not well documented. This study evaluates the cytotoxic effects and mechanism of action of EO extracts on MCF-7 and Vero cell lines. Evaluation of the cytotoxic activity using MTT assay, morphological alterations, and apoptosis were carried out. Autophagy was evaluated by LC3-A protein expression. Cytotoxic activity, membrane blebbing and ballooning at 24 hours, replacement by mass vacuolation, and double membrane vesicles mimicking autophagy and cell death were observed in the cancer cells. No apoptosis was observed by DNA fragmentation assay. Overexpression of LC3-A protein indicated autophagic cell death. Cell cycle analysis showed G0 and G2/M arrest. The Vero cells did not show significant cell death at concentrations <100??g/mL. These results thus suggest that acetone and ethyl acetate extracts of EO induce cell death through induction of autophagy and hold potential for development as potential anticancer drugs.

Harun, Faizah Bt.; Syed Sahil Jamalullail, Syed Mohsin; Yin, Khoo Boon; Othman, Zulkhairi; Tilwari, Anita; Balaram, Prabha

2012-01-01

63

Preclinical evaluation of Vaxfectin-adjuvanted Vero cell-derived seasonal split and pandemic whole virus influenza vaccines.  

PubMed

Increasing the potency and supply of seasonal and pandemic influenza vaccines remains an important unmet medical need which may be effectively accomplished with adjuvanted egg- or cell culture-derived vaccines. Vaxfectin, a cationic lipid-based adjuvant with a favorable safety profile in phase 1 plasmid DNA vaccines trials, was tested in combination with seasonal split, trivalent and pandemic whole virus, monovalent influenza vaccines produced in Vero cell cultures. Comparison of hemagglutination inhibition (HI) antibody titers in Vaxfectin-adjuvanted to nonadjuvanted vaccinated mice and guinea pigs revealed 3- to 20-fold increases in antibody titers against each of the trivalent influenza virus vaccine strains and 2- to 8-fold increases in antibody titers against the monovalent H5N1 influenza virus vaccine strain. With the vaccine doses tested, comparable antibody responses were induced with formulations that were freshly prepared or refrigerated at conventional 2-8°C storage conditions for up to 6 mo. Comparison of T-cell frequencies measured by interferon-gamma ELISPOT assay between groups revealed increases of between 2- to 10-fold for each of the adjuvanted trivalent strains and up to 22-fold higher with monovalent H5N1 strain. Both trivalent and monovalent vaccines were easy to formulate with Vaxfectin by simple mixing. These preclinical data support further testing of Vaxfectin-adjuvanted Vero cell culture vaccines toward clinical studies designed to assess safety and immunogenicity of these vaccines in humans. PMID:23857272

Smith, Larry R; Wodal, Walter; Crowe, Brian A; Kerschbaum, Astrid; Bruehl, Peter; Schwendinger, Michael G; Savidis-Dacho, Helga; Sullivan, Sean M; Shlapobersky, Mark; Hartikka, Jukka; Rolland, Alain; Barrett, P Noel; Kistner, Otfried

2013-06-01

64

Preclinical evaluation of Vaxfectin®-adjuvanted Vero cell-derived seasonal split and pandemic whole virus influenza vaccines  

PubMed Central

Increasing the potency and supply of seasonal and pandemic influenza vaccines remains an important unmet medical need which may be effectively accomplished with adjuvanted egg- or cell culture-derived vaccines. Vaxfectin®, a cationic lipid-based adjuvant with a favorable safety profile in phase 1 plasmid DNA vaccines trials, was tested in combination with seasonal split, trivalent and pandemic whole virus, monovalent influenza vaccines produced in Vero cell cultures. Comparison of hemagglutination inhibition (HI) antibody titers in Vaxfectin®-adjuvanted to nonadjuvanted vaccinated mice and guinea pigs revealed 3- to 20-fold increases in antibody titers against each of the trivalent influenza virus vaccine strains and 2- to 8-fold increases in antibody titers against the monovalent H5N1 influenza virus vaccine strain. With the vaccine doses tested, comparable antibody responses were induced with formulations that were freshly prepared or refrigerated at conventional 2–8°C storage conditions for up to 6 mo. Comparison of T-cell frequencies measured by interferon-gamma ELISPOT assay between groups revealed increases of between 2- to 10-fold for each of the adjuvanted trivalent strains and up to 22-fold higher with monovalent H5N1 strain. Both trivalent and monovalent vaccines were easy to formulate with Vaxfectin® by simple mixing. These preclinical data support further testing of Vaxfectin®-adjuvanted Vero cell culture vaccines toward clinical studies designed to assess safety and immunogenicity of these vaccines in humans.

Smith, Larry R.; Wodal, Walter; Crowe, Brian A.; Kerschbaum, Astrid; Bruehl, Peter; Schwendinger, Michael G.; Savidis-Dacho, Helga; Sullivan, Sean M.; Shlapobersky, Mark; Hartikka, Jukka; Rolland, Alain; Barrett, P. Noel; Kistner, Otfried

2013-01-01

65

Characterization and detection of Vero cells infected with Herpes Simplex Virus type 1 using Raman spectroscopy and advanced statistical methods.  

PubMed

Herpes viruses are involved in a variety of human disorders. Herpes Simplex Virus type 1 (HSV-1) is the most common among the herpes viruses and is primarily involved in human cutaneous disorders. Although the symptoms of infection by this virus are usually minimal, in some cases HSV-1 might cause serious infections in the eyes and the brain leading to blindness and even death. A drug, acyclovir, is available to counter this virus. The drug is most effective when used during the early stages of the infection, which makes early detection and identification of these viral infections highly important for successful treatment. In the present study we evaluated the potential of Raman spectroscopy as a sensitive, rapid, and reliable method for the detection and identification of HSV-1 viral infections in cell cultures. Using Raman spectroscopy followed by advanced statistical methods enabled us, with sensitivity approaching 100%, to differentiate between a control group of Vero cells and another group of Vero cells that had been infected with HSV-1. Cell sites that were "rich in membrane" gave the best results in the differentiation between the two categories. The major changes were observed in the 1195-1726cm(-1) range of the Raman spectrum. The features in this range are attributed mainly to proteins, lipids, and nucleic acids. PMID:24582780

Salman, A; Shufan, E; Zeiri, L; Huleihel, M

2014-07-01

66

Expression of the newcastle disease virus fusion glycoprotein in vero cells using attenuated Salmonella typhimurium as transgenic carrier.  

PubMed

The full-length cDNA of fusion protein (F) gene of newcastle disease virus (NDV)strain F48E9 was amplified by RT-PCR and inserted into pcDNA3 under the control of human cytomegalovirus (hCMV) immediate early enhancer and promoter. The resulting recombinant plasmid pcDNA3-F was transformed by electroporation into attenuated Salmonella typhimurium strain ZJ111 (dam(-) and phoP(-)), which was then used to transfect the Vero cells. The DNA and RNA dot blotting revealed that the F gene was transcribed into mRNA in the Vero cells. There was expression of the F protein as shown by indirect immunofluorescent assay. The expression began at 48 h post-infection and increased thereafter, as indicated by ELISA. A 55 kD band of the F protein was identified by SDS-PAGE and Western blotting. These results clearly show that the expressed fusion protein was immuno-reactive with chicken anti-NDV serum. PMID:12098773

Fang, Wei-Huan; Liang, Xue-Ya

2002-07-01

67

Growth and poliovirus production of Vero cells on a novel microcarrier with artificial cell adhesive protein under serum-free conditions.  

PubMed

A microcarrier is used for the three-dimensional (3D) culture of adhesion-dependent mammalian cells. We developed a novel microcarrier by binding ProNectin F, an artificial cell adhesive protein synthesized by genetically engineered Escherichia coli to a polyacrylic superabsorbent polymer. The microcarrier is characterized by containing no animal-derived components. The serum-free culture of Vero cells for vaccine production using the microcarrier increased the number of Vero cells by approximately 30% compared with the existing dextran beads coated with porcine Type I collagen, which resulted in approximately a 30% to 40% increase in the infectivity titer of the Sabin 2 strain of poliovirus. These results suggested that the developed microcarrier should be unprecedented in permitting high-yield vaccine production by means of a serum-free culture. PMID:21262586

Kurokawa, Masato; Sato, Shigehiro

2011-05-01

68

Differentiation of a Vero cell adapted porcine epidemic diarrhea virus from Korean field strains by restriction fragment length polymorphism analysis of ORF 3.  

PubMed

A porcine epidemic diarrhea virus (PEDV) designated DR13 was isolated in Vero cells and serially passaged by level 100. The virus was titrated at regular intervals of the passage level. Open reading frame (ORF) 3 sequences of the virus at passage levels 20, 40, 60, 80, and 100 were aligned and compared using a computer software program. Suitability of the restriction fragment length polymorphism (RFLP) analysis for differentiating the virus from other Korean field strains was investigated. The DR13 field isolate was successively adapted in Vero cells as observed through polymerase chain reaction (PCR) and titration of the virus. RFLP analysis identified change in cleavage sites of HindIII and Xho II from passage levels 75 and 90, respectively; these RFLP patterns of ORF 3 differentiated the Vero cell-adapted virus from its parent strain, DR13, and 12 other strains of PEDV studied. The cell adapted DR13 was tested for its pathogenicity and immunogenicity in piglets and pregnant sows. The results indicated that cell adapted DR13 revealed reduced pathogenicity and induced protective immune response in pigs. Differentiation between highly Vero cell-adapted virus and wild-type virus could be the marker of adaptation to cell culture and a valuable tool for epidemiologic studies of PEDV infections. The results of this study supported that the cell attenuated virus could be applied as a marker vaccine candidate against PEDV infection. PMID:12706667

Song, D S; Yang, J S; Oh, J S; Han, J H; Park, B K

2003-05-16

69

Enhanced Growth of Influenza Vaccine Seed Viruses in Vero Cells Mediated by Broadening the Optimal pH Range for Virus Membrane Fusion  

PubMed Central

Vaccination is one of the most effective preventive measures to combat influenza. Prospectively, cell culture-based influenza vaccines play an important role for robust vaccine production in both normal settings and urgent situations, such as during the 2009 pandemic. African green monkey Vero cells are recommended by the World Health Organization as a safe substrate for influenza vaccine production for human use. However, the growth of influenza vaccine seed viruses is occasionally suboptimal in Vero cells, which places limitations on their usefulness for enhanced vaccine production. Here, we present a strategy for the development of vaccine seed viruses with enhanced growth in Vero cells by changing an amino acid residue in the stem region of the HA2 subunit of the hemagglutinin (HA) molecule. This mutation optimized the pH for HA-mediated membrane fusion in Vero cells and enhanced virus growth 100 to 1,000 times in the cell line, providing a promising strategy for cell culture-based influenza vaccines.

Murakami, Shin; Ito, Mutsumi; Takano, Ryo; Katsura, Hiroaki; Shimojima, Masayuki

2012-01-01

70

[Vitaherpavac is the first Russian herpes simplex virus vaccine obtained on the Vero B continuous cell line].  

PubMed

Vitaherpavac, a dry inactivated herpes simplex virus (HSV) culture vaccine, has been obtained, by using the Vero B continuous cell line as a substrate for accumulation of herpes simplex virus types 1 (US strain) and 2 (VN strain). Vitaherpavac and the similar vaccine Herpovax made by the Research Institute of Vaccines and Sera, Saint Petersburg (for which preparation a primary trypsinized chick embryo cell culture used as a substrate for accumulation of HSV types 1 and 2), underwent comparative clinical trials. The tolerability and therapeutic effectiveness of the vaccine were tested in patients diagnosed as having chronic frequently recurring herpes. The trials have yielded positive results that suggest that it is expedient to introduce of the new vaccine Vitaherpavac into practice to treat chronic recurrent herpetic infection of various localizations. Vitaherpavac has been registered in the Russian Federation and permitted for medical application. PMID:19882901

Barkhaleva, O A; Ladyzhenskaia, I P; Vorob'eva, M S; Shalunova, N V; Podcherniaeva, R Ia; Mikha?lova, G R; Khorosheva, T V; Barinski?, I F

2009-01-01

71

Isolation of measles virus from clinical specimens using B95a and Vero/hSLAM cell-lines.  

PubMed

The clinical presentation of acute measles is normally quite typical, especially in the presence of Koplik's spots, that laboratory test is seldom required to confirm the diagnosis. However, with wide measles vaccination coverage and the extensive use of immunosuppressive chemotherapy, the diagnosis of atypical manifestations of acute measles may require laboratory confirmation. When compared with B95a cell-line, this study shows that the Vero/hSLAM cell-line is sensitive and is recommended for use in the primary isolation of wild-type measles virus from clinical specimens. Throat swab and urine specimens are the clinical specimens of choice and both are recommended for optimal isolation of measles virus from patients suspected of acute measles virus infection. PMID:19852319

Keniscope, C; Juliana, R; Subri, H; Shangari, S R; Wan Nor Azlina, W A; Hamizah, A; Emmi, E E; Nor Azlina, M D; Norizah, I; Chua, K B

2009-03-01

72

Apoptosis induced in an early step of African swine fever virus entry into vero cells does not require virus replication.  

PubMed

Permissive Vero cells develop apoptosis, as characterized by DNA fragmentation, caspases activation, cytosolic release of mitochondrial cytochrome c, and flow cytometric analysis of DNA content, upon infection with African swine fever virus (ASFV). To determine the step in virus replication that triggers apoptosis, we used UV-inactivated virus, inhibitors of protein and nucleic acid synthesis, and lysosomotropic drugs that block virus uncoating. ASFV-induced apoptosis was accompanied by caspase-3 activation, which was detected even in the presence of either cytosine arabinoside or cycloheximide, indicating that viral DNA replication and protein synthesis were not required to activate the apoptotic process. The activation of caspase-3 was released from chloroquine inhibition 2 h after virus absorption, while the infection with UV-inactivated ASFV did not induce the activation of the caspase cascade. We conclude that ASFV induces apoptosis in the infected cell by an intracellular pathway probably triggered during the process of virus uncoating. PMID:12009879

Carrascosa, Angel L; Bustos, María J; Nogal, María L; González de Buitrago, Gonzalo; Revilla, Yolanda

2002-03-15

73

Non-Linear Relationships between Aflatoxin B1 Levels and the Biological Response of Monkey Kidney Vero Cells  

PubMed Central

Aflatoxin-producing fungi contaminate food and feed during pre-harvest, storage and processing periods. Once consumed, aflatoxins (AFs) accumulate in tissues, causing illnesses in animals and humans. Most human exposure to AF seems to be a result of consumption of contaminated plant and animal products. The policy of blending and dilution of grain containing higher levels of aflatoxins with uncontaminated grains for use in animal feed implicitly assumes that the deleterious effects of low levels of the toxins are linearly correlated to concentration. This assumption may not be justified, since it involves extrapolation of these nontoxic levels in feed, which are not of further concern. To develop a better understanding of the significance of low dose effects, in the present study, we developed quantitative methods for the detection of biologically active aflatoxin B1 (AFB1) in Vero cells by two independent assays: the green fluorescent protein (GFP) assay, as a measure of protein synthesis by the cells, and the microculture tetrazolium (MTT) assay, as a measure of cell viability. The results demonstrate a non-linear dose-response relationship at the cellular level. AFB1 at low concentrations has an opposite biological effect to higher doses that inhibit protein synthesis. Additional studies showed that heat does not affect the stability of AFB1 in milk and that the Vero cell model can be used to determine the presence of bioactive AFB1 in spiked beef, lamb and turkey meat. The implication of the results for the cumulative effects of low amounts of AFB1 in numerous foods is discussed.

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

2013-01-01

74

Non-linear relationships between aflatoxin B? levels and the biological response of monkey kidney vero cells.  

PubMed

Aflatoxin-producing fungi contaminate food and feed during pre-harvest, storage and processing periods. Once consumed, aflatoxins (AFs) accumulate in tissues, causing illnesses in animals and humans. Most human exposure to AF seems to be a result of consumption of contaminated plant and animal products. The policy of blending and dilution of grain containing higher levels of aflatoxins with uncontaminated grains for use in animal feed implicitly assumes that the deleterious effects of low levels of the toxins are linearly correlated to concentration. This assumption may not be justified, since it involves extrapolation of these nontoxic levels in feed, which are not of further concern. To develop a better understanding of the significance of low dose effects, in the present study, we developed quantitative methods for the detection of biologically active aflatoxin B? (AFB1) in Vero cells by two independent assays: the green fluorescent protein (GFP) assay, as a measure of protein synthesis by the cells, and the microculture tetrazolium (MTT) assay, as a measure of cell viability. The results demonstrate a non-linear dose-response relationship at the cellular level. AFB1 at low concentrations has an opposite biological effect to higher doses that inhibit protein synthesis. Additional studies showed that heat does not affect the stability of AFB1 in milk and that the Vero cell model can be used to determine the presence of bioactive AFB1 in spiked beef, lamb and turkey meat. The implication of the results for the cumulative effects of low amounts of AFB1 in numerous foods is discussed. PMID:23949006

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

2013-08-01

75

Comparison of N-Glycan Pattern of Recombinant Human Coagulation Factors II and IX Expressed in Chinese Hamster Ovary (CHO) and African Green Monkey (Vero) Cells.  

PubMed

The N-glycan patterns of recombinant human coagulation factors II (rF-II) and IX (rF-IX), derived from both transfected Chinese hamster ovary (CHO) cells and African green monkay (Vero) cells produced at industrial scale, were analyzed by binding to carbohydrate-specific lectins and were compared with the glycan structure of human plasma-derived coagulation factors. Human plasma-derived coagulation factors II (hpF-II) and IX (hpF-IX) exhibited complex-type glycan structures with carbohydrate chains capped with alpha(2-6)-sialic acid. Terminal galactose-beta(1-4)-N-acetylglucosamine units were detected in hpF-IX. Both CHO cell-derived rF-II and rF-IX exhibited complex-type glycosylation and contained alpha(2-3)-sialic acid in addition to terminal galactose-beta(1-4)-N-acetylglucosamine. Vero cell-derived rF-IX exhibited a complex-type glycan structure similar to that of CHO cell-derived rF-IX. In contrast, rF-II produced by Vero cells exhibited a glycan microheterogeneity composed of hybrid-type glycosylation containing "high-mannose" structures and complex-type glycosylation containing alpha(2-3)-sialic acid. Galactose-beta(1-4)-N-acetylglucosamine structures and a low concentration of alpha(2-6)-sialic acid were detected in both microheterogeneity fractions of Vero cell-derived rF-II. Although different in their carbohydrate structures, coagulation factors II and IX obtained recombinantly from both transformed CHO cells and Vero cells exhibited coagulation activities comparable with the plasma-derived proteins. PMID:10608038

Fischer; Mitterer; Dorner; Eibl

1996-01-01

76

A/H5N1 prepandemic influenza vaccine (whole virion, vero cell-derived, inactivated) [Vepacel®].  

PubMed

The influenza A subtype H5N1 virus is a likely causative agent for the next human influenza pandemic. Pandemic influenza vaccine production can begin only after a novel pandemic virus emerges. Cell-based vaccine production has advantages over conventional egg-based methods, allowing more rapid large-scale vaccine production. A reliable Vero cell culture system is available for pandemic and prepandemic influenza vaccine production. Prepandemic influenza vaccines are an important component of influenza pandemic preparedness plans, as their targeted use in the pandemic alert period or early in a pandemic is likely to mitigate the consequences of an influenza outbreak. Vepacel® is a prepandemic influenza vaccine (whole virion, Vero cell-derived, inactivated) containing antigen of H5N1 strain A/Vietnam/1203/2004 and is approved for use in the EU. Clinical immunogenicity studies with the vaccine have demonstrated good rates of functional neutralizing antibody responses against the vaccine strain (A/Vietnam/1203/2004), meeting established immunogenicity criteria for seasonal influenza vaccines, and cross-reactivity against H5N1 strains from other clades. In phase I/II and III studies, a heterologous (A/Indonesia/05/2005) booster vaccine administered to healthy adult and elderly volunteers 6-24 months after the two-dose priming vaccine (A/Vietnam/1203/2004) regimen induced good immunogenic responses against both H5N1 strains, demonstrating strong immunological memory. Broadly similar, albeit less robust, responses were observed in two special risk cohorts of immunocompromised and chronically ill patients. In general, adverse events observed in clinical immunogenicity studies with H5N1 vaccine (A/Vietnam/1203/2004) were similar to those reported with non-adjuvanted, inactivated, seasonal influenza vaccines. PMID:22788239

Plosker, Greg L

2012-07-30

77

Inhibitory effect of oryzacystatins and a truncation mutant on the replication of poliovirus in infected Vero cells.  

PubMed

Poliovirus, a picornavirus family member, requires the processing of its poly-protein by its own cysteine proteinase for replication. Oryzacystatin-I and oryzacystatin-II, proteinaceous cysteine proteinase inhibitors (cystatins) of rice seed origin, were found to inhibit the replication of poliovirus effectively in infected Vero cells in vitro. Truncated oryzacystatin-I, which lacks the NH2-terminal 25 amino acid residues of the intact protein, is an even more effective inhibitor, eliciting its effect at concentrations of less than 0.25 nmol/ml. The low molecular weight cysteine proteinase inhibitors, E-64, E-64C and loxistatin, showed no anti-viral effect at any concentration investigated. PMID:1312033

Kondo, H; Ijiri, S; Abe, K; Maeda, H; Arai, S

1992-03-24

78

Autophagic cell death is induced by acetone and ethyl acetate extracts from Eupatorium odoratum in vitro: effects on MCF-7 and vero cell lines.  

PubMed

Eupatorium odoratum (EO) contains many biologically active compounds, the anticancer effects of which are not well documented. This study evaluates the cytotoxic effects and mechanism of action of EO extracts on MCF-7 and Vero cell lines. Evaluation of the cytotoxic activity using MTT assay, morphological alterations, and apoptosis were carried out. Autophagy was evaluated by LC3-A protein expression. Cytotoxic activity, membrane blebbing and ballooning at 24 hours, replacement by mass vacuolation, and double membrane vesicles mimicking autophagy and cell death were observed in the cancer cells. No apoptosis was observed by DNA fragmentation assay. Overexpression of LC3-A protein indicated autophagic cell death. Cell cycle analysis showed G0 and G2/M arrest. The Vero cells did not show significant cell death at concentrations <100??g/mL. These results thus suggest that acetone and ethyl acetate extracts of EO induce cell death through induction of autophagy and hold potential for development as potential anticancer drugs. PMID:22666123

Harun, Faizah Bt; Syed Sahil Jamalullail, Syed Mohsin; Yin, Khoo Boon; Othman, Zulkhairi; Tilwari, Anita; Balaram, Prabha

2012-01-01

79

Isolation of bovine coronavirus (bcoV) in vero cell line and its confirmation by direct FAT and RT-PCR.  

PubMed

Bovine Coronavirus (BCoV) is widespread both in dairy and beef cattle throughout the world. The virus is one of the largest RNA virus and has specific tropism for intestinal and pulmonary epithelial cells. It is responsible for huge economic losses by causing winter dysentery in adult dairy cattle and respiratory and intestinal tract infections leading to pneumo-enteritis in young calves. Isolation of BCoV has been reported to be difficult. Studies regarding epidemiology, virus isolation and molecular detection from India are very few. In the present study Vero cell line was used for isolation of the BCoV from Enzyme Linked Immunosorbent Assay (ELISA) positive samples. Direct florescent antibody technique (dFAT) and reverse transcriptase-polymerase chain reaction (RT-PCR) were used to confirm the isolated virus strains at antigenic and genomic levels, respectively. Out of the 15 positive fecal samples, virus from only seven was able to infect vero cell line. Subsequently BCoV got adapted to the vero cell line upto three passages, which was confirmed both at genomic and antigenic levels by dFAT and RT-PCR testing. It can be concluded that vero cell line can be used for isolation of BCoV, however due to the enormous stain diversity of the virus it is possible that many stains can't grow and get adapt in this cell line. Further studies are required for isolation of different viral strains, finding the susceptible cell lines and also to confirm the variations among the BCoV isolates at antigenic/genomic levels. PMID:24511744

Hansa, A; Rai, R B; Dhama, K; Wani, M Y; Saminathan, M; Ranganath, G J

2013-11-01

80

Purification of Vero cell derived live replication deficient influenza A and B virus by ion exchange monolith chromatography.  

PubMed

We explored the possibilities for purification of various ?NS1 live, replication deficient influenza viruses on ion exchange methacrylate monoliths. Influenza A ?NS1-H1N1, ?NS1-H3N2, ?NS1-H5N1 and ?NS1-influenza B viruses were propagated in Vero cells and concentrated by tangential flow filtration. All four virus strains adsorbed well to CIM QA and CIM DEAE anion exchangers, with CIM QA producing higher recoveries than CIM DEAE. ?NS1-influenza A viruses adsorbed well also to CIM SO3 cation exchanger at the same pH, while ?NS1-influenza B virus adsorption to CIM SO3 was not complete. Dynamic binding capacity (DBC) for CIM QA, DEAE and SO3 methacrylate monoliths for influenza A ?NS1-H1N1 virus were 1.9E+10 TCID50/ml, 1.0E+10 TCID50/ml and 8.9E+08 TCID50/ml, respectively. Purification of ?NS1 viruses on CIM QA was scaled up and reproducibility was confirmed. Yields of infectious virus on CIM QA were between 70.8±32.3% and 87±30.8%. Total protein removal varied from 93.3±0.4% to 98.6±0.2% and host cell DNA removal efficiency was ranging from 76.4% to 99.9% and strongly depended on pretreatment with deoxyribonuclease. PMID:24631091

Banjac, Marko; Roethl, Elisabeth; Gelhart, Franz; Kramberger, Petra; Jarc, Barbara Lah; Jarc, Marko; Strancar, Aleš; Muster, Thomas; Peterka, Matjaž

2014-05-01

81

The major outer membrane protein rOmpB of spotted fever group rickettsiae functions in the rickettsial adherence to and invasion of Vero cells  

Microsoft Academic Search

The role of one of the major outer membrane proteins, rOmpB, of spotted fever group rickettsiae was examined. Antibodies generated against native rOmpB inhibited plaque formation by Rickettsia japonica in Vero cells when applied at the time of inoculation of the rickettsiae. However, antibodies to heat-denatured rOmpB did not. Moreover, the soluble recombinant rOmpB also inhibited plaque formation to some

Tsuneo Uchiyama; Hiroaki Kawano; Yoshito Kusuhara

2006-01-01

82

Safety and Immunogenicity of Inactivated, Vero Cell Culture-Derived Whole Virus Influenza A/H5N1 Vaccine Given Alone or with Aluminum Hydroxide Adjuvant in Healthy Adults  

PubMed Central

Dosage-sparing strategies, adjuvants and alternative substrates for vaccine production are being explored for influenza vaccine development. We assessed the safety and immunogenicity of a Vero cell culture-grown inactivated whole virus influenza A/H5N1 vaccine with or without aluminum hydroxide adjuvant [Al(OH)3] in healthy young adults. Vaccines were well tolerated, but injection site discomfort was more frequent in groups receiving Al(OH)3. Dose-related increases in serum antibody levels were observed. Neutralizing antibody titers varied significantly when tested by two different laboratories. Al(OH)3 did not enhance HAI or neutralizing antibody responses, and contributed to increased injection site pain. Because influenza antibody titers vary significantly between different laboratories, international standardization of assays is warranted.

Keitel, Wendy A.; Dekker, Cornelia L.; Mink, ChrisAnna; Campbell, James D.; Edwards, Kathryn M.; Patel, Shital M.; Ho, Dora Y.; Talbot, Helen K.; Guo, Kuo; Noah, Diana L.; Hill, Heather

2011-01-01

83

A vero cell-derived whole-virus H5N1 vaccine effectively induces neuraminidase-inhibiting antibodies.  

PubMed

A Vero cell-derived whole-virus H5N1 influenza vaccine has been shown to induce neutralizing antibodies directed against the hemagglutinin (HA) protein of diverse H5N1 strains in animal studies and clinical trials. However, neuraminidase-inhibiting (NAi) antibodies can reduce viral spread and may be of particular importance in the event of an H5N1 pandemic, where immunity due to HA antibodies is likely absent in the general population. Here we demonstrate the effective induction of NAi antibody titers after H5N1 vaccination in humans. In contrast to the immune response directed toward HA, a single vaccine dose induced a strong NAi response that was not significantly boosted by a second dose, most probably due to priming by previous vaccination or infection with seasonal influenza viruses. After 2 immunizations, seroconversion rates based on antibody titers against HA and NA were similar, indicating the induction of equally strong immune responses against both proteins by this H5N1 vaccine. PMID:22090447

Fritz, Richard; Sabarth, Nicolas; Kiermayr, Stefan; Hohenadl, Christine; Howard, M Keith; Ilk, Reinhard; Kistner, Otfried; Ehrlich, Hartmut J; Barrett, P Noel; Kreil, Thomas R

2012-01-01

84

Purification, potency and immunogenicity analysis of Vero cell culture-derived rabies vaccine: a comparative study of single-step column chromatography and zonal centrifuge purification.  

PubMed

Continuous Vero cell lines are more suitable for large-scale production of rabies vaccine. The purification of Vero cell-derived rabies vaccine is critical because of the residual cellular DNA and serum proteins. The perfection of techniques using column chromatography with different matrix material, gel filtration and zonal centrifugation is of paramount importance for the optimal purification of rabies vaccine, leaving minimal residual cellular DNA, below the permissible level of 100 pg per dose and serum protein content of 1 ppm. In this study the potency, immunogenicity and safety of Vero cell-derived rabies vaccines were compared following purification by densely or loosely packed DEAE-sepharose CL-6B columns with different bed heights or by zonal centrifugation. The optimal virus recovery and maximum removal of substrate DNA and serum proteins were achieved only when the sepharose CL-6B column bed height was maintained at a thickness of 2-2.5 cm. The rabies virus material was purified by layering over the matrix without applying pressure. DEAE-sepharose CL-6B column purification using a simplified, cost effective technique as described in this study enhances the antigen yield by 50% in comparison with zonal purification. PMID:16046167

Prem Kumar, Ananda Arone; Mani, Kavaratty Raju; Palaniappan, Chitrambalam; Bhau, Lakshman Narasingha Rao; Swaminathan, Krishnaswami

2005-07-01

85

Immunogenicity and protective efficacy in mice of influenza B virus vaccines grown in mammalian cells or embryonated chicken eggs.  

PubMed

The immunogenicity and protective efficacy of formalin-inactivated influenza B/Memphis/1/93 virus vaccines propagated exclusively in Vero cells, MDCK cells, or embryonated chicken eggs (hereafter referred to as eggs) were investigated. Mammalian cell-grown viruses differ from the egg-grown variant at amino acid position 198 (Pro/Thr) in the hemagglutinin gene. The level of neuraminidase activity was highest in egg-grown virus, while MDCK and Vero cell-derived viruses possessed 70 and 90% less activity, respectively. After boosting, each of the vaccines induced high levels of hemagglutinin-inhibiting, neuraminidase-inhibiting, and neutralizing antibodies that provided complete protection from MDCK-grown virus challenge. Mammalian cell-derived virus vaccines induced serum antibodies that were more cross-reactive, while those induced by egg-grown virus vaccines were more specific to the homologous antigen. Enzyme-linked immunospot analysis indicated that cell-grown virus vaccines induced high frequencies of immunoglobulin G (IgG)-producing cells directed against both cell- and egg-grown virus antigens, whereas egg-grown virus vaccine induced higher frequencies of IgG- and IgM-producing cells reacting with homologous antigen and low levels of IgG-producing cells reactive with cell-grown viruses. These studies indicate that influenza B virus variants selected in different host systems can elicit different immune responses, but these alterations had no detectable influence on the protective efficacy of the vaccines with the immunization protocol used in this study. PMID:9557744

Alymova, I V; Kodihalli, S; Govorkova, E A; Fanget, B; Gerdil, C; Webster, R G

1998-05-01

86

Immunogenicity and Protective Efficacy in Mice of Influenza B Virus Vaccines Grown in Mammalian Cells or Embryonated Chicken Eggs  

PubMed Central

The immunogenicity and protective efficacy of formalin-inactivated influenza B/Memphis/1/93 virus vaccines propagated exclusively in Vero cells, MDCK cells, or embryonated chicken eggs (hereafter referred to as eggs) were investigated. Mammalian cell-grown viruses differ from the egg-grown variant at amino acid position 198 (Pro/Thr) in the hemagglutinin gene. The level of neuraminidase activity was highest in egg-grown virus, while MDCK and Vero cell-derived viruses possessed 70 and 90% less activity, respectively. After boosting, each of the vaccines induced high levels of hemagglutinin-inhibiting, neuraminidase-inhibiting, and neutralizing antibodies that provided complete protection from MDCK-grown virus challenge. Mammalian cell-derived virus vaccines induced serum antibodies that were more cross-reactive, while those induced by egg-grown virus vaccines were more specific to the homologous antigen. Enzyme-linked immunospot analysis indicated that cell-grown virus vaccines induced high frequencies of immunoglobulin G (IgG)-producing cells directed against both cell- and egg-grown virus antigens, whereas egg-grown virus vaccine induced higher frequencies of IgG- and IgM-producing cells reacting with homologous antigen and low levels of IgG-producing cells reactive with cell-grown viruses. These studies indicate that influenza B virus variants selected in different host systems can elicit different immune responses, but these alterations had no detectable influence on the protective efficacy of the vaccines with the immunization protocol used in this study.

Alymova, I. V.; Kodihalli, S.; Govorkova, E. A.; Fanget, B.; Gerdil, C.; Webster, R. G.

1998-01-01

87

Bovine colostrum ultrafiltrate supplemented with adult bovine serum and transferrin: An effective fbs substitute for cultivation of vero and CHO-K1 cells  

Microsoft Academic Search

Summary  A mixture containing an ultrafiltrate fraction (UF) of bovine colostrum (6.7%), adult bovine serum (BS) (1%), and human holo-transferrin\\u000a (hTF) (5 mg\\/liter) was developed for cultivation of Chinese hamster ovary cells (CHO-K1) and African green monkey kidney cells\\u000a (Vero). The growth-supporting activity of the mixture (UF\\/BS\\/hTF) was comparable to that of 1 to 10% fetal bovine serum (FBS)\\u000a and considerably

Raimo Pakkanen

1994-01-01

88

Induction of indistinguishable gene expression patterns in rats by Vero cell-derived and mouse brain-derived Japanese encephalitis vaccines.  

PubMed

Transcriptomics is an objective index that reflects the overall condition of cells or tissues, and transcriptome technology, such as DNA microarray analysis, is now being introduced for the quality control of medical products. In this study, we applied DNA microarray analysis to evaluate the character of Japanese encephalitis (JE) vaccines. When administered into rat peritoneum, Vero cell-derived and mouse brain-derived JE vaccines induced similar gene expression patterns in liver and brain. Body weights and blood biochemical findings were also similar after administration of the two vaccines. Our results suggest that the two JE vaccines are likely to have equivalent characteristics with regard to reactivity in rats. PMID:20093758

Momose, Haruka; Imai, Jun-ichi; Hamaguchi, Isao; Kawamura, Mika; Mizukami, Takuo; Naito, Seishiro; Masumi, Atsuko; Maeyama, Jun-ichi; Takizawa, Kazuya; Kuramitsu, Madoka; Nomura, Nobuo; Watanabe, Shinya; Yamaguchi, Kazunari

2010-01-01

89

Reduction of spiked porcine circovirus during the manufacture of a Vero cell-derived vaccine.  

PubMed

Porcine circovirus-1 (PCV1) was recently identified as a contaminant in live Rotavirus vaccines, which was likely caused by contaminated porcine trypsin. The event triggered the development of new regulatory guidance on the use of porcine trypsin which shall ensure that cell lines and porcine trypsin in use are free from PCV1. In addition, manufacturing processes of biologicals other than live vaccines include virus clearance steps that may prevent and mitigate any potential virus contamination of product. In this work, artificial spiking of down-scaled models for the manufacturing process of an inactivated pandemic influenza virus vaccine were used to investigate inactivation of PCV1 and the physico-chemically related porcine parvovirus (PPV) by formalin and ultraviolet-C (UV-C) treatment as well as removal by the purification step sucrose gradient ultracentrifugation. A PCV1 infectivity assay, using a real-time PCR infectivity readout was established. The formalin treatment (0.05% for 48h) showed substantial inactivation for both PCV1 and PPV with reduction factors of 3.0log10 and 6.8log10, respectively, whereas UV-C treatment resulted in complete PPV (?5.9log10) inactivation already at a dose of 13mJ/cm but merely 1.7log10 at 24mJ/cm(2) for PCV1. The UV-C inactivation results with PPV were confirmed using minute virus of mice (MVM), indicating that parvoviruses are far more sensitive to UV-C than PCV1. The sucrose density gradient ultracentrifugation also contributed to PCV1 clearance with a reduction factor of 2log10. The low pH treatment during the production of procine trypsin was investigated and showed effective inactivation for both PCV1 (4.5log10) and PPV (6.4log10). In conclusion, PCV1 in general appears to be more resistant to virus inactivation than PPV. Still, the inactivated pandemic influenza vaccine manufacturing process provides for robust virus reduction, in addition to the already implemented testing for PCV1 to avoid any contaminations. PMID:24560672

Lackner, Cornelia; Leydold, Sandra M; Modrof, Jens; Farcet, Maria R; Grillberger, Leopold; Schäfer, Birgit; Anderle, Heinz; Kreil, Thomas R

2014-04-11

90

Aggravation of cold-induced injury in Vero-B4 cells by RPMI 1640 medium - Identification of the responsible medium components  

PubMed Central

Background In modern biotechnology, there is a need for pausing cell lines by cold storage to adapt large-scale cell cultures to the variable demand for their products. We compared various cell culture media/solutions for cold storage of Vero-B4 kidney cells, a cell line widely used in biotechnology. Results Cold storage in RPMI 1640 medium, a recommended cell culture medium for Vero-B4 cells, surprisingly, strongly enhanced cold-induced cell injury in these cells in comparison to cold storage in Krebs-Henseleit buffer or other cell culture media (DMEM, L-15 and M199). Manufacturer, batch, medium supplements and the most likely components with concentrations outside the range of the other media/solutions (vitamin B12, inositol, biotin, p-aminobenzoic acid) did not cause this aggravation of cold-induced injury in RPMI 1640. However, a modified Krebs-Henseleit buffer with a low calcium concentration (0.42 mM), a high concentration of inorganic phosphate (5.6 mM), and glucose (11.1 mM; i.e. concentrations as in RPMI 1640) evoked a cell injury and loss of metabolic function corresponding to that observed in RPMI 1640. Deferoxamine improved cell survival and preserved metabolic function in modified Krebs-Henseleit buffer as well as in RPMI 1640. Similar Ca2+ and phosphate concentrations did not increase cold-induced cell injury in the kidney cell line LLC-PK1, porcine aortic endothelial cells or rat hepatocytes. However, more extreme conditions (Ca2+ was nominally absent and phosphate concentration raised to 25 mM as in the organ preservation solution University of Wisconsin solution) also increased cold-induced injury in rat hepatocytes and porcine aortic endothelial cells. Conclusion These data suggest that the combination of low calcium and high phosphate concentrations in the presence of glucose enhances cold-induced, iron-dependent injury drastically in Vero-B4 cells, and that a tendency for this pathomechanism also exists in other cell types.

2012-01-01

91

Induction of micronuclei by Zearalenone in Vero monkey kidney cells and in bone marrow cells of mice: protective effect of Vitamin E.  

PubMed

Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin mainly produced by Fusarium graminaerum, found as a world-wide contaminant mainly of corn and wheat. Previous studies have demonstrated that among several other effects on animals and humans, ZEN also displays hepatotoxicity, immunotoxicity and nephrotoxicity. ZEN is mainly known as a hormonal disrupter due to its estrogenic activities and consequent toxicity for reproduction. Furthermore, mutagenic and genotoxic proprieties of ZEN were disclosed recently, the molecular mechanisms of which are not yet well understood. In the present study, the genotoxic potential of ZEN was evaluated using genotoxicity tests: the 'cytokinesis block micronucleus assay' in Vero monkey kidney cells and the 'in vivo mouse bone marrow micronucleus assay'. In cultured cells treated with 5, 10 and 20 microM ZEN, the frequency of binucleated micronucleated cells (BNMN) was assessed in 1000 binucleated cells and in mice given oral doses of 10, 20 and 40 mg/kg bw, the frequency of polychromatic erythrocytes micronucleated (PCEMN) in bone marrow cells was assessed in 2000 polychromatic erythrocytes (PCE). The potential prevention of ZEN-induced effects by 25 microM Vitamin E (Vit E) was also evaluated. In vivo, doses of 10, 20 and 40 mg/kg bw ZEN representing, respectively 2, 4 and 8% of the LD50 (LD50 of ZEN in mice is 500 mg/kg bw), were administered to animals either with or without pre-treatment with Vit E (216.6 mg/kg bw) in order to evaluate its preventive potential.ZEN was found to induce micronuclei (MN) in a dose-dependent manner in cultured Vero cells as well as in mouse bone marrow cells. The present data emphasise the likely clastogenic pathway among the molecular mechanisms that underlay the ZEN-induced genotoxicity. Vit E was found to prevent partially-from 30 to 50%-these toxic effects, most likely acting either as a structural analogue of ZEN or as an antioxidant. PMID:12834755

Ouanes, Zouhour; Abid, Salwa; Ayed, Imen; Anane, Rachid; Mobio, Théophile; Creppy, Edmond E; Bacha, Hassen

2003-07-01

92

Comparison of human immune responses to purified Vero cell and human diploid cell rabies vaccines by using two different antibody titration methods.  

PubMed Central

Antibody responses to a conventional rabies preexposure regimen of a new purified Vero cell rabies vaccine (PVRV) and a human diploid cell vaccine (HDCV) were compared in 80 healthy Kenyan veterinary students. Forty-three of the students received the PVRV and 37 received the HDCV on days 0, 7, and 28. Antibody responses were monitored by using the rapid fluorescent-focus inhibition test (RFFIT) and an inhibition enzyme immunoassay (INH EIA) on days 0, 7, 28, and 49. Both vaccines elicited a rapid antibody response. A good correlation between the RFFIT titers and the INH EIA titers was obtained (r = 0.90). Our results also showed that the INH EIA was more reproducible and might therefore be a suitable substitute for the more expensive and less reproducible RFFIT. The geometric mean titers determined by both tests in the two groups of students were statistically similar during the test period. The RFFIT and the INH EIA gave comparable geometric mean titers, which differed significantly only on day 28 in the PVRV group. The effect of the new PVRV is comparable to that of the more expensive HDCV, as determined by the present test systems. The PVRV could therefore be the vaccine of choice, especially in tropical rabies-endemic areas, where the high cost of the HDCV has confined its use to a privileged few.

Kitala, P M; Lindqvist, K J; Koimett, E; Johnson, B K; Chunge, C N; Perrin, P; Olsvik, O

1990-01-01

93

The influence of serum substituents on serum-free Vero cell conditioned culture media manufactured from Dulbecco's modified Eagle medium in mouse embryo culture  

PubMed Central

Objective This study was conducted to examine the influences of supplementation of the serum substituents and available period of serum-free Vero cell conditioned media (SF-VCM) manufactured from Dulbecco's modified Eagle medium cultured with Vero cells for in vitro development of mouse preimplantation embryos. Methods A total of 1,099 two-cell embryos collected from imprinting control region mice were cultured in SF-VCM with 10% and 20% human follicular fluid (hFF), serum substitute supplement (SSS), and serum protein substitute (SPS). Development of embryos was observed every 24 hours. Results between different groups were analyzed by chi-square test, and considered statistically significant when P-value was less than 0.05. Results The rates of embryonic development cultured in SF-VCM supplemented with serum substituents were significantly higher compare with serum-free group (P < 0.05). The rates of embryonic development after 48 hours (morula?) and 96 hours (blastocyst?) were significantly higher in 20% SSS and 10% SPS than in 20% hFF supplementation (P < 0.05). And the rates of embryonic development after 96 hours (hatching blastocyst?) were significantly higher in 10% SPS (94.5%) than in 20% SSS (82.6%) and 20% hFF supplementation (68.5%). The rates of embryonic development according to storage period of the SF-VCM supplemented with 10% SPS showed no significant difference between control, 2 weeks and 4 weeks group. However developmental rate in 6 weeks storage group was significantly lower than other groups. Conclusion The rate of embryonic development after 96 hours (hatching blastocyst?) was significantly higher in SF-VCM supplemented with 10% SPS. And storage period of media up to 4 weeks did not affect on embryonic development.

Lee, Jong-Seon; Kim, Ju-Hwan; Seo, Young-Seok; Yang, Jung-Bo; Kim, Yong-Il; Kim, Hye-Jin

2013-01-01

94

Comparison of procedures for the extraction of supernatants and cytotoxicity tests in Vero cells, applied to assess the toxigenic potential of Bacillus spp. and Lactobacillus spp., intended for use as probiotic strains.  

PubMed

Interest in using Bacillus strains as probiotic components of animal feeds has grown in recent years. However, some of these strains, especially those taxonomically related to the Bacillus cereus group, may have enterotoxigenic activity. Assessment of their toxigenic potential by well-established and robust protocols is required before authorizing their use in animal nutrition. Three methods of extraction and concentration of supernatants of Bacillus and Lactobacillus strains (methanol extraction, ammonium sulphate and ultrafiltration concentration) and three cytotoxic tests in Vero cells (WST-1, LDH and protein synthesis inhibition assays) for the assessment of the cytotoxicity activity of Lactobacillus strains (as probiotic strains in human and animal nutrition) and Bacillus toyonensis BCT-7112(T) (as animal probiotic strain in animal nutrition-Toyocerin®-) were evaluated in this study. Methanol extraction was not useful under any circumstances. The other two concentration methods (ammonium sulphate and ultrafiltration) were feasible, with slightly greater sensitivity achieved by ultrafiltration. The probiotic strain B. toyonensis BCT-7112(T) proved to be a non-cytotoxic strain in all the protocols tested. However, some Lactobacillus strains showed cytotoxicity activity, regardless of the protocols applied. PMID:24938520

Blanch, Anicet R; Méndez, Javier; Castel, Susana; Reina, Manuel

2014-08-01

95

Morphological analysis of the transfer of VSV ts-045 G glycoprotein from the endoplasmic reticulum to the intermediate compartment in vero cells.  

PubMed

Vero cells were infected with the ts-045 strain of vesicular stomatitis virus, and the cells were incubated at 39 degrees C to accumulate the mutant G glycoprotein in the ER as a misfolded aggregate. Cycloheximide was added to the culture medium 3.5 h after infection to prevent further protein synthesis, and the temperature was lowered to 10, 15, or 31 degrees C. At these temperatures, the mutant G glycoprotein correctly folds and oligomerizes. Immunofluorescence light microscopy showed that the G glycoprotein was exported to the Golgi complex at 31 degrees C and to the intermediate compartment (IC) at 15 degrees C, but no export was observed at 10 degrees C. However, incubations at 10 degrees C followed by shift to 15 or 31 degrees C resulted in the normal transfer of the glycoprotein to the IC and the Golgi, respectively. Immunoelectron microscopical analysis confirmed all these results, but showed also that the glycoprotein was frequently clustered in the ER at 10 degrees C. Conventional electron microscopy showed that the morphology of the ER, IC, and Golgi complex remained essentially unchanged at all temperatures. The only significant difference detectable in cells incubated at 10 degrees C was the increased number of partially coated ER protrusions, longer than those detected at higher temperatures. These results demonstrate that the transport toward the Golgi complex of G glycoprotein can be arrested at a step preceding the entry into the IC, thus suggesting that ER and IC are separate stations in the exocytic pathway. PMID:8831570

Lotti, L V; Torrisi, M R; Erra, M C; Bonatti, S

1996-09-15

96

Inhibition of cytotoxicity of Shiga toxin of Escherichia coli O157:H7 on vero cells by Prosopis alba Griseb (Fabaceae) and Ziziphus mistol Griseb (Rhamnaceae) extracts.  

PubMed

The capacity of Prosopis alba Griseb. and Ziziphus mistol Griseb. fruit extracts to inhibit the toxic action of Shiga toxin (Stx) was investigated. Purification of Stx from Escherichia coli O157:H7 was performed by saline precipitation and affinity chromatography using a column with globotriaosylceramide, while the fruits were subjected to ethanolic or aqueous extractions. The protective action of both fruits was determined by pre-, co-, and postincubation of one 50% cytotoxic dose per ml of Stx with different concentrations of ethanolic and aqueous extracts in confluent monolayers of Vero cells for 72 h at 37°C (5% CO2). The inhibition of the cytotoxic effect of Stx by fruit extracts was determined by the neutral red vital staining technique. The extraction of the polyphenols and flavonoids was effective, and more polyphenols per milligram of dissolved solids were obtained from P. alba than from Z. mistol. However, there were more flavonoids in Z. mistol than in P. alba. Components of both fruits increased the viability of cells treated with Stx when the extracts were preincubated with Stx for 1 h before being applied to the cell cultures, with the ethanolic extract of P. alba showing 95% cell viability at a concentration of 2.45 mg/ml. The extracts were less effective in protecting cells when Stx, extracts, and cells were coincubated together without a previous incubation of Stx; only the concentrations of 19.46 mg/ml for the P. alba aqueous extract and 3.75 mg/ml for the Z. mistol ethanolic extract resulted in the inhibition of cytotoxicity, with 52 and 56% cell viability occurring, respectively. Investigation into this difference in the protection of cells indicated that the protein molecule of Stx suffered degradation to advanced oxidative protein products during preincubation with extracts, principally with P. alba, which exhibited a greater amount of nonflavonoid polyphenols than Z. mistol. The prooxidant action on Stx favored the cells and enhanced the protective action of both fruits. PMID:24112573

Pellarín, M G; Albrecht, C; Rojas, M J; Aguilar, J J; Konigheim, B S; Paraje, M G; Albesa, I; Eraso, A J

2013-10-01

97

Colon tumor cells grown in NASA Bioreactor  

NASA Technical Reports Server (NTRS)

These photos compare the results of colon carcinoma cells grown in a NASA Bioreactor flown on the STS-70 Space Shuttle in 1995 flight and ground control experiments. The cells grown in microgravity (left) have aggregated to form masses that are larger and more similar to tissue found in the body than the cells cultured on the ground (right). The principal investigator is Milburn Jessup of the University of Texas M. D. Anderson Cancer Center. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. Cell constructs grown in a rotating bioreactor on Earth (left) eventually become too large to stay suspended in the nutrient media. In the microgravity of orbit, the cells stay suspended. Rotation then is needed for gentle stirring to replenish the media around the cells. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). Credit: NASA and University of Texas M. D. Anderson Cancer Center.

2001-01-01

98

Safety analysis of a Vero-cell culture derived Japanese encephalitis vaccine, IXIARO (IC51), in 6 months of follow-up.  

PubMed

Japanese encephalitis (JE) is the most common viral encephalitis in Asia. IXIARO is a Vero cell-derived, inactivated JE virus vaccine which has recently been approved in the US, Europe, Canada and Australia (trade name JESPECT). This overview of the safety and tolerability of IXIARO, for 6 months after the first vaccination in 7 Phase III trials, includes: 3558 subjects with at least one IXIARO vaccination, 435 subjects with a JE-VAX (manufactured by BIKEN, distributed by Sanofi Pasteur) vaccination, and 657 with phosphate-buffered saline solution with 0.1% Al(OH)(3) (PBS+Alum) control vaccination. The percentage of subjects reporting solicited local adverse events (AEs) with IXIARO (54%) was similar to PBS+Alum vaccination (56%) as were solicited systemic adverse events (40% IXIARO; 40% PBS+Alum vaccination). JE-VAX showed a higher frequency of subjects with solicited local adverse events (61%) but a slightly lower frequency of subjects with solicited systemic adverse events (36%). The frequency of subjects with any solicited and unsolicited AE with IXIARO (64%) was also similar to PBS+Alum vaccination (61%) and JE-VAX (64%); as for subjects with serious AEs (1% IXIARO; 2% PBS+Alum vaccination, 1% JE-VAX). No serious allergic reactions were observed in any group. This safety analysis indicates that IXIARO has a favorable safety profile, comparable to PBS+Alum control vaccination and appears to have a better local tolerability profile than JE-VAX. PMID:20673824

Dubischar-Kastner, Katrin; Kaltenboeck, Astrid; Klingler, Anton; Jilma, Bernd; Schuller, Elisabeth

2010-09-01

99

The amino-terminal one-third of pseudorabies virus glycoprotein gIII contains a functional attachment domain, but this domain is not required for the efficient penetration of Vero cells.  

PubMed Central

We have examined the attachment and penetration phenotypes of several glycoprotein gIII mutants of pseudorabies virus (PRV) and have identified the first one-third of gIII as a region that mediates efficient virus attachment to PK15 and Vero cells. This portion of gIII, amino acids 25 through 157 of the wild-type sequence, appeared to support attachment by binding to heparinlike molecules on cell surfaces. Virions containing the first one-third of gIII were sensitive to heparin competition and showed greatly reduced infectivity on cells treated with heparinase. PRV virions lacking the first one-third of the mature glycoprotein exhibited only residual binding to cells if challenged by vigorous washing with phosphate-buffered saline at 2 h postinfection at 4 degrees C. This residual binding was resistant to heparin competition, and strains lacking the first one-third of gIII were able to infect cells treated with heparinase as effectively as untreated cells. When we determined the penetration phenotypes for each strain, we found that gIII-mediated virus attachment was necessary for timely penetration of PK15 cells but remarkably was not required for efficient virus penetration of Vero cells. Moreover, wild-type PRV was actually prohibited from rapid penetration of Vero cells by a gIII-heparan sulfate interaction. Our results indicate that initial virus binding to heparan sulfate via glycoprotein gIII is not required for efficient PRV infection of all cell types and may in fact be detrimental in some instances. Images

Flynn, S J; Burgett, B L; Stein, D S; Wilkinson, K S; Ryan, P

1993-01-01

100

Process standardization for optimal virus recovery and removal of substrate DNA and bovine serum proteins in Vero cell-derived rabies vaccine.  

PubMed

Purification of a rabies vaccine by a single zonal centrifugation run was replaced by two runs with optimal standardization of the sucrose density gradient. As a result, significant reductions in the levels of substrate DNA and bovine serum protein in the Vero cell-derived human rabies vaccine were achieved. Following many trials, for the first run, loading of the 3.2-l capacity K-3 rotor with 1800 ml of 60% sucrose solution and 1400 ml of vaccine PBS buffer solution gave a satisfactory linear gradient. However, after the first run, the substrate DNA and bovine serum contents exceeded the required levels. After protamine sulphate and Tween-80 treatment of the concentrated inactivated material, a second run using the same procedure as in the first run was tried. However, these purification procedures resulted in low virus recovery. To achieve optimal virus recovery, and removal of substrate DNA and bovine serum protein, the peak fractions from the first run as indicated by the haemagglutination, sucrose concentration, and optical density values were pooled and the sucrose concentration of the pooled fractions was increased to 60%. A second (flotation) run was then carried out. Using this method, the virus recovery rate was more than 95% that of the first run, and the levels of cellular DNA and bovine serum protein were well within the acceptable limits of less than 100 pg/dose and one part per million, respectively. The substrate DNA was quantified by both radioactive labeling and non-radioactive biotin labeling methods. For the quantification of calf serum protein, a counter-immunoelectrophoresis method was developed and effectively applied. A potency assay was performed using the National Institutes of Health (NIH) and well-standardized in vitro single radial immuno diffusion (SRD) methods. Finally, an immunogenicity study was conducted with human volunteers and the results were confirmed by a rapid fluorescent focus inhibition test (RFFIT). PMID:16233321

Kumar, Ananda Arone Prem; Rao, Yarlagadda Udaya Bhaskara; Joseph, Arokiaswami Leo William; Mani, Kavaratty Raju; Swaminathan, Krishnaswami

2002-01-01

101

Characterization of yellow fever virus proteins E and NS1 expressed in Vero and Spodoptera frugiperda cells.  

PubMed

The cDNA encoding the E and NS1 proteins of the yellow fever virus (YFV) was expressed in Spodoptera frugiperda cells via the recombinant baculovirus Ac-E. NS1 as a gp100 precursor which was cleaved to generate the recombinant proteins E and NS1 similar in size, folding and antigenicity to the authentic ones. Recombinant protein E exhibited immunodominant epitopes as judged by its reactivity with YFV-neutralizing MAbs. Using the Triton X-114 phase separation system, authentic and recombinant E proteins as well as the gp100 precursor exhibited hydrophobic properties similar to those of integral membrane proteins. Recombinant protein E was found neither in the extracellular medium nor on the cell surface, suggesting that it did not migrate within the secretory pathway of insect cells. Analysis of protein NS1 expressed in primate and insect cells revealed that the newly synthesized 48K NS1 glycoprotein was converted to a heat-labile gp72 homo-oligomeric form. This phenomenon did not require the presence of carbohydrate groups. Using the Triton X-114 phase separation system, the oligomeric form of NS1 was shown to be associated with cellular membranes although it appeared less hydrophobic than protein E and gp100. A small fraction of YFV NS1 oligomers were transported throughout the secretory pathway to be shed into the extracellular medium of primate cells. YFV NS1 oligomers migrated from the endoplasmic reticulum to the Golgi complex, whereas their N-oligosaccharides of the high-mannose type are processed to the complex-mannose type. Protein NS1 expressed by recombinant baculovirus-infected insect cells was not found in the extracellular medium but associated with the plasma membrane of the cells. Two recombinant NS1 forms were detected in insect cells: a major one with an apparent Mr of 48K and a minor one of 47K in which N-linked glycans were probably processed to a trimannosyl core without further elongation. Thus, it appears that the transport strategy as well as the N-glycosylation of NS1 in insect cells infected with recombinant baculovirus were different from those of the NS1 in primate cells infected with YFV. PMID:1710649

Desprès, P; Girard, M; Bouloy, M

1991-06-01

102

Comparative study on the cytotoxicity of different Myrtaceae essential oils on cultured vero and RC-37 cells.  

PubMed

Medicinally and commercially important essential oils from the family Myrtaceae, i.e. cajuput, clove, kanuka and manuka were phytochemically analysed by GC-MS. Cytotoxicity of these essential oils was evaluated in a standard neutral red assay. Maximum noncytotoxic concentrations for cajuput oil and clove oil were determined at 0.006%, kanuka oil and manuka oil were more cytotoxic with a maximum noncytotoxic concentration of 0.001%. The compounds alpha-pinene, eugenol and leptospermone demonstrated maximum noncytotoxic concentrations at dilutions of 0.001%, 0.003% and 0.001%, respectively. However, the terpene 1,8-cineole was about 100 times less toxic to cultured cells with a maximum noncytotoxic concentration of 0.1% and a TC50 value of 0.44%. Manuka essential oil exhibited high levels of virucidal activity against HSV-1 as well against drug-resistant HSV-1 isolates in viral suspension tests. Determination of cytotoxicity of natural products is an important prerequisite for application in cosmetic and health care products and in antiviral tests. PMID:19069246

Schnitzler, P; Wiesenhofer, K; Reichling, J

2008-11-01

103

The specific activities of Shiga-like toxin type II (SLT-II) and SLT-II-related toxins of enterohemorrhagic Escherichia coli differ when measured by Vero cell cytotoxicity but not by mouse lethality.  

PubMed Central

Characteristically, enterohemorrhagic Escherichia coli (EHEC) strains produce Shiga-like toxin type I (SLT-I), SLT-II, or both of these immunologically distinct cytotoxins. No antigenic or receptor-binding variants of SLT-I have been identified, but a number of SLT-II-related toxins have been described. Because EHEC O91:H21 strain B2F1, which produces two SLT-II-related toxins, is exquisitely virulent in an orally infected, streptomycin-treated mouse model (oral 50% lethal dose [LD50], < 10 organisms), we asked whether the pathogenicity of strain B2F1 was a consequence of SLT-II-related toxin production. For this purpose, we compared the lethality of orally administered E. coli DH5 alpha (Strr) strains that produced different cytotoxic levels of SLT-II, SLT-IIvha (cloned from B2F1), SLT-IIvhb (also cloned from B2F1), or SLT-IIc (cloned from EHEC O157:H7 strain E32511) on Vero cells. We also calculated the specific activities of purified SLT-IIvhb and SLT-II in intraperitoneally injected mice and on Vero cells. The two purified toxins were equally toxic for mice, but SLT-IIvhb was approximately 100-fold less active than SLT-II on Vero cells and bound to the glycolipid receptor Gb3 with lower affinity than did SLT-II. In addition, characterization of SLT-II-related toxin-binding (B) subunit mutants generated in this study revealed that the reduced in vitro cytotoxic levels of the SLT-II-related toxins were due to Asn-16 in the B subunit. Taken together, these findings do not support the idea that B2F1 is uniquely virulent because of the in vivo toxicity of SLT-II-related toxins but do demonstrate differences in in vitro cytotoxic activity among the SLT-II group produced by human EHEC isolates. Images

Lindgren, S W; Samuel, J E; Schmitt, C K; O'Brien, A D

1994-01-01

104

A Single Dose of Vero Cell-Derived Japanese Encephalitis (JE) Vaccine (Ixiaro) Effectively Boosts Immunity in Travelers Primed With Mouse Brain-Derived JE Vaccines  

PubMed Central

Background.?A significant part of the world population lives in areas with endemic Japanese encephalitis (JE). For travelers from nonendemic countries, Vero cell–derived vaccine (JE-VC; Ixiaro) has replaced traditional mouse brain–derived vaccines (JE-MB) associated with safety concerns. The 2 vaccines are derived from different viral strains: JE-VC from the SA14-14-2 strain and JE-MB from the Nakayama strain. No data exist regarding whether JE-VC can be used to boost immunity after a primary series of JE-MB; therefore, a primary series of JE-VC has been recommended to all travelers regardless of previous vaccination history. Methods.?One hundred twenty travelers were divided into 4 groups: Volunteers with no prior JE vaccination received primary immunization with (group 1) JE-MB or (group 2) JE-VC, and those primed with JE-MB received a single booster dose of (group 3) JE-MB or (group 4) JE-VC. Immune responses were tested before and 4–8 weeks after vaccination using plaque reduction neutralization test (PRNT) against both vaccine strains. Results.?In vaccine-naive travelers, the vaccination response rate for test strains Nakayama and SA14-14-2 was 100% and 87% after primary vaccination with JE-MB and 87% and 94% after JE-VC, respectively. Antibody levels depended on the target virus, with higher titers against homologous than heterologous PRNT50 target strain (P < .001). In travelers primed with JE-MB, vaccination response rates were 91% and 91%, and 98% and 95% after a booster dose of JE-MB or JE-VC, respectively. Subgroup analysis revealed that a higher proportion of primed (98%/95%) than nonprimed (39%/42%) volunteers responded to a single dose of JE-VC (P < .001). Conclusions.?A single dose of JE-VC effectively boosted immunity in JE-MB–primed travelers. Current recommendations should be reevaluated. Clinical Trials Registration.?NCT01386827.

Erra, Elina O.; Askling, Helena Hervius; Rombo, Lars; Riutta, Jukka; Vene, Sirkka; Yoksan, Sutee; Lindquist, Lars; Pakkanen, Sari H.; Huhtamo, Eili; Vapalahti, Olli; Kantele, Anu

2012-01-01

105

Vero cell-derived inactivated West Nile (WN) vaccine induces protective immunity against lethal WN virus infection in mice and shows a facilitated neutralizing antibody response in mice previously immunized with Japanese encephalitis vaccine.  

PubMed

A novel Vero cell-derived inactivated WN vaccine (WN-VAX) was prepared from virus strain NY99-35262. Two immunizations with WN-VAX induced high levels of neutralizing antibody to WN virus. All immunized mice were protected against challenge with a lethal dose of WN virus. No WN viremia was detected, and the level of WN virus-neutralizing antibody increased rapidly. WN-VAX was then examined for immunogenicity in mice previously immunized with Japanese encephalitis vaccine (JE-VAX). Immunization with WN-VAX induced WN virus-neutralizing antibody in all mice previously immunized with JE-VAX but in only half of the control mice at 10 weeks. These results indicate that WN-VAX induced complete protective immunity against lethal WN infection and that the WN-VAX-induced antibody response is facilitated in JE-VAX-immunized mice. This WN-VAX is thus a candidate WN vaccine for humans. PMID:18221765

Lim, Chang-Kweng; Takasaki, Tomohiko; Kotaki, Akira; Kurane, Ichiro

2008-04-25

106

Long-term immunogenicity of the new Vero cell-derived, inactivated Japanese encephalitis virus vaccine IC51 Six and 12 month results of a multicenter follow-up phase 3 study.  

PubMed

Japanese encephalitis (JE) is the most common viral encephalitis in Asia. IC51 is a new Vero cell-derived, inactivated JE virus vaccine with non-inferior immunogenicity (after 2 months) compared to the US-licensed vaccine JE-VAX (mouse brain-derived, inactivated) and with a more convenient (two injections instead of three) intramuscular dose schedule. Adult subjects from two studies were followed-up for comparative immunogenicity (JE-VAX) at 6 months and long-term immunogenicity of IC51 alone at 12 months. At 6 months, immunogenicity was higher with IC51 (seroconversion rate [SCR] 95%; geometric mean titer [GMT] 84) than with JE-VAX (SCR 74%; GMT 34). At 12 months, the SCR was 83% and the GMT (41) remained above the protective titer of 1:10. Most people immunized with IC51 will have protective neutralizing antibody levels for at least a year. PMID:18599165

Schuller, E; Jilma, B; Voicu, V; Golor, G; Kollaritsch, H; Kaltenböck, A; Klade, C; Tauber, E

2008-08-12

107

Quantitative Proteomics Using Stable Isotope Labeling with Amino Acids in Cell Culture Reveals Changes in the Cytoplasmic, Nuclear, and Nucleolar Proteomes in Vero Cells Infected with the Coronavirus Infectious Bronchitis Virus*  

PubMed Central

Virus-host interactions involve complex interplay between viral and host factors, rendering them an ideal target for proteomic analysis. Here we detail a high throughput quantitative proteomics analysis of Vero cells infected with the coronavirus infectious bronchitis virus (IBV), a positive strand RNA virus that replicates in the cytoplasm. Stable isotope labeling with amino acids in cell culture (SILAC) was used in conjunction with LC-MS/MS to identify and quantify 1830 cellular and two viral proteins from IBV-infected cells. Fractionation of cells into cytoplasmic, nuclear, and nucleolar extracts was used to reduce sample complexity and provide information on the trafficking of proteins between the different compartments. Each fraction showed a proportion of proteins exhibiting ?2-fold changes in abundance. Ingenuity Pathway Analysis revealed that proteins that changed in response to infection could be grouped into different functional categories. These included proteins regulated by NF-?B- and AP-1-dependent pathways and proteins involved in the cytoskeleton and molecular motors. A luciferase-based reporter gene assay was used to validate the up-regulation of AP-1- and NF-?B-dependent transcription in IBV-infected cells and confirmed using immunofluorescence. Immunofluorescence was used to validate changes in the subcellular localization of vimentin and myosin VI in IBV-infected cells. The proteomics analysis also confirmed the presence of the viral nucleocapsid protein as localizing in the cytoplasm, nucleus, and nucleolus and the viral membrane protein in the cytoplasmic fraction. This research is the first application of SILAC to study total host cell proteome changes in response to positive sense RNA virus infection and illustrates the versatility of this technique as applied to infectious disease research.

Emmott, Edward; Rodgers, Mark A.; Macdonald, Andrew; McCrory, Sarah; Ajuh, Paul; Hiscox, Julian A.

2010-01-01

108

Gene expression in Fusarium graminearum grown on plant cell wall  

Microsoft Academic Search

Fusarium graminearum is a phytopathogenic filamentous fungus attacking a wide range of plants including Humulus lupulus (hop). Transcriptional analysis of F. graminearum grown on minimal media containing hop cell wall or glucose as the sole carbon source was performed by applying a highly stringent method combining microarrays and a subtracted cDNA library. In addition to genes coding for various cell

Raphaël Carapito; Didier Hatsch; Sonja Vorwerk; Elizabet Petkovski; Jean-Marc Jeltsch; Vincent Phalip

2008-01-01

109

Stem Cells Yield Lab-Grown Skin, Researchers Say  

MedlinePLUS

... sharing features on this page, please enable JavaScript. Stem Cells Yield Lab-Grown Skin, Researchers Say Finding might ... April 25, 2014 Related MedlinePlus Pages Skin Conditions Stem Cells FRIDAY, April 25, 2014 (HealthDay News) -- Skin that ...

110

Pandemic influenza A H1N1 vaccine in recipients of solid organ transplants: immunogenicity and tolerability outcomes after vero cell derived, non-adjuvanted, whole-virion vaccination.  

PubMed

During the 2009/10 pandemic of influenza A (H1N1), the American Society of Transplantation and other health organizations recommended that immunocompromised patients should be vaccinated as the key preventive measure. Since there are no data available for the immunogenicity of the unadjuvanted pandemic influenza vaccine in immunocompromised patients - as opposed to the adjuvanted preparation - the objective of this study was to evaluate the immunogenicity of an adjuvant-free H1N1 vaccine in recipients of solid organ transplants. Patients were recruited at the Vienna General Hospital, Austria. The vaccination schedule consisted of 2 doses of a whole-virion, vero cell derived, inactivated, non-adjuvanted influenza A/California/07/2009 (H1N1) vaccine given with an interval of 3 weeks. A hemagglutination inhibition (HI) assay on blood samples obtained prior to the first and after each vaccination was used for serologic analysis. The primary immunologic endpoint was the seroconversion rate, defined as the proportion of subjects with an individual 4-fold increase in HI titer of at least 1:40. In addition, virus-specific IgG antibodies to the pandemic H1N1 strain were measured using a commercially available ELISA. Twenty-five organ transplant patients (16 males, 9 females) aged 25-79 years were vaccinated and provided blood samples for serologic analysis. The time elapsed since transplantation was 10 months to 25 years (mean: 9 years; 95% CI 6-13 years). The vaccine was well tolerated and no local adverse events were noticed. After two vaccinations 37% of the patients demonstrated seroconversion in the HI assay as defined above and 70% had virus-specific IgG antibodies. Among the HI vaccine responders were 6 of 14 heart transplant recipients and 1 of 4 liver transplant recipients. The number and type of immunosuppressive agents did not significantly differ in their effect on the immune response. Our results show that the novel vero cell derived and adjuvant-free pandemic A/California/07/2009 (H1N1) influenza vaccine induced limited but measurable immune responses in adult recipients of solid organ transplants. PMID:21803100

Lagler, Heimo; Wenisch, Judith M; Tobudic, Selma; Gualdoni, Guido A; Rödler, Susanne; Rasoul-Rockenschaub, Susanne; Jaksch, Peter; Redlberger-Fritz, Monika; Popow-Kraupp, Theresia; Burgmann, Heinz

2011-09-16

111

Heteroepitaxially grown InP solar cells  

Microsoft Academic Search

The properties of InP solar cells, processed by OMCVD on silicon substrates with an intermediate GaAs layer (InP\\/GaAs\\/Si) and on GaAs substrates (InP\\/GaAs), were determined before and after irradiation with 10-MeV protons. The preirradiation transport properties were found to be influenced largely by dislocations occurring at the InP-GaAs interface. A carrier removal rate of 1.8×103 cm-1 was observed after irradiation

I. Weinberg; C. K. Swartz; D. J. Brinker; D. M. Wilt

1990-01-01

112

Comparison of vero cell plaque assay, TaqMan reverse transcriptase polymerase chain reaction RNA assay, and VecTest antigen assay for detection of West Nile virus in field-collected mosquitoes.  

PubMed

Mosquitoes collected during the epidemic of West Nile virus (WN) in Staten Island, NY, during 2000 were identified to species, grouped into pools of up to 50 individuals, and tested for the presence of WN by using TaqMan reverse transcriptase polymerase chain reaction (RT-PCR) to detect West Nile viral RNA, Vero cell plaque assay to detect infectious virus, and VecTest WNV/SLE Antigen Panel Assay. A total of 10,866 specimens was tested in 801 pools. Analysis of results indicated that TaqMan RT-PCR detected 34 WN-positive pools, more than either of the other techniques. The plaque assay detected 74% of the pools positive by TaqMan, and VecTest detected 60% of the pools positive by TaqMan. The VecTest assay detected evidence of West Nile viral antigen in 67% of the pools that contained live virus detected by plaque assay. A WN enzyme immunoassay performed similarly to the VecTest WN assay. Differences in performance were related to relative sensitivity of the tests. Infection rates of WN in Culex pipiens and Cx. salinarius calculated by the 3 techniques varied, but each estimate indicated a high infection rate in the population. Positive and negative attributes of each procedure, which may influence how and where they are used in surveillance programs, are discussed. PMID:12542186

Nasci, Roger S; Gottfried, Kristy L; Burkhalter, Kristen L; Kulasekera, Varuni L; Lambert, Amy J; Lanciotti, Robert S; Hunt, Ann R; Ryan, Jeffrey R

2002-12-01

113

OM-VPE grown materials for high efficiency solar cells  

NASA Technical Reports Server (NTRS)

Organometallic sources are available for all the III-V elements and a variety of dopants; thus it is possible to use the technique to grow a wide variety of semiconductor compounds. AlGaAsSb and AlGaInAs alloys for multijunction monolithic solar cells were grown by OM-VPE. While the effort concentrated on terrestrial applications, the success of OM-VPE grown GaAs/AlGaAs concentrator solar cells (23% at 400 suns) demonstrates that OM-VPE is suitable for growing high efficiency solar cells in large quantities for space applications. In addition, OM-VPE offers the potential for substantial cost reduction of photovoltaic devices with scale up and automation and due to high process yield from reproducible, uniform epitaxial growths with excellent surface morphology.

Saxena, R.; Cooper, B., III; Ludowise, M.; Borden, P.; Gregory, P.

1980-01-01

114

Safety and Immunogenicity of a Vero Cell Culture-Derived Whole-Virus H5N1 Influenza Vaccine in Chronically Ill and Immunocompromised Patients.  

PubMed

The development of vaccines against H5N1 influenza A viruses is a cornerstone of pandemic preparedness. Clinical trials of H5N1 vaccines have been undertaken in healthy subjects, but studies in risk groups have been lacking. In this study, the immunogenicity and safety of a nonadjuvanted cell culture-derived whole-virus H5N1 vaccine were assessed in chronically ill and immunocompromised adults. Subjects received two priming immunizations with a clade 1 A/Vietnam H5N1 influenza vaccine, and a subset also received a booster immunization with a clade 2.1 A/Indonesia H5N1 vaccine 12 to 24 months later. The antibody responses in the two populations were assessed by virus neutralization and single radial hemolysis assays. The T-cell responses in a subset of immunocompromised patients were assessed by enzyme-linked immunosorbent spot assay (ELISPOT). The priming and the booster vaccinations were safe and well tolerated in the two risk populations, and adverse reactions were predominantly mild and transient. The priming immunizations induced neutralizing antibody titers of ?1:20 against the A/Vietnam strain in 64.2% of the chronically ill and 41.5% of the immunocompromised subjects. After the booster vaccination, neutralizing antibody titers of ?1:20 against the A/Vietnam and A/Indonesia strains were achieved in 77.5% and 70.8%, respectively, of chronically ill subjects and in 71.6% and 67.5%, respectively, of immunocompromised subjects. The T-cell responses against the two H5N1 strains increased significantly over the baseline values. Substantial heterosubtypic T-cell responses were elicited against the 2009 pandemic H1N1 virus and seasonal A(H1N1), A(H3N2), and B subtypes. There was a significant correlation between T-cell responses and neutralizing antibody titers. These data indicate that nonadjuvanted whole-virus cell culture-derived H5N1 influenza vaccines are suitable for immunizing chronically ill and immunocompromised populations. (This study is registered at ClinicalTrials.gov under registration no. NCT00711295.). PMID:24739978

van der Velden, Maikel V W; Geisberger, Alexander; Dvorak, Thomas; Portsmouth, Daniel; Fritz, Richard; Crowe, Brian A; Herr, Wolfgang; Distler, Eva; Wagner, Eva M; Zeitlinger, Markus; Sauermann, Robert; Stephan, Christoph; Ehrlich, Hartmut J; Barrett, P Noel; Aichinger, Gerald

2014-06-01

115

Safety and immunogenicity of two different doses of a Vero cell-derived, whole virus clade 2 H5N1 (A/Indonesia/05/2005) influenza vaccine.  

PubMed

A successful vaccine development strategy for areas with clustered H5N1 events requires conduct of vaccine trials in potentially non-naïve subjects and evaluation of post-vaccination responsiveness. An open-label, randomized, phase I/II study therefore assessed the immunogenicity and safety of two different dose levels of an inactivated, non-adjuvanted, whole virus clade 2.1 (A/Indonesia/05/2005) H5N1 Vero cell-derived influenza vaccine in healthy adults (21-45 years) from a region where the virus has been circulating (Hong Kong) as well as Singapore. Subjects (N=110) were randomized 1:1 to receive two vaccinations with either 3.75 ?g or 7.5 ?g H5N1 haemagglutinin antigen 21 days apart. Safety, immunogenicity (microneutralization [MN] and single radial haemolysis [SRH] at baseline and post-vaccination) and cross-reactivity against a heterologous clade 1 strain (A/Vietnam/1203/2004) of the vaccine were assessed. Pre-existing immunity to the vaccine strain was 14% which is higher than previously reported for these regions. Two vaccinations with either vaccine formulation induced high seroprotection rates (MN titre ? 1:20) against the vaccine strain A/Indonesia/05/2005: 82.7% and 86.5% in the 3.75 ?g and 7.5 ?g dose groups. Seroconversion rates and fold increase exceeded the CPMP criterion of >40% and >2.5 for MN and SRH in both dose groups after the second vaccination, while the seroprotection rate in the 7.5 ?g dose group determined by SRH was only marginally lower (69.2%) than the CPMP criterion of >70%. Thus, 11 of 12 CHMP criteria were fulfilled. A cross-reactive antibody response against the heterologous A/Vietnam/1203/2004 strain was demonstrated after the second vaccination (>21% by MN and ? 25% by SRH). Persistence of antibodies against the vaccine strain was also demonstrated 6 months after the first vaccination, indicating that a booster vaccination would be effective in those who have received two priming doses. No serious adverse events were reported. The H5N1 influenza vaccine against clade 2.1 strain A/Indonesia/05/2005 was well tolerated and immunogenic after two vaccinations, and induced a cross-neutralizing antibody response, with no dose effect. PMID:22080174

Tambyah, Paul A; Wilder-Smith, Annelies; Pavlova, Borislava G; Barrett, P Noel; Oh, Helen M L; Hui, David S; Yuen, Kwok-yung; Fritsch, Sandor; Aichinger, Gerald; Loew-Baselli, Alexandra; van der Velden, Maikel; Maritsch, Friedrich; Kistner, Otfried; Ehrlich, Hartmut J

2012-01-01

116

Replication of Borna disease virus in cell cultures.  

PubMed

Borna disease (BD) virus from infected brain tissue of horses or rabbits could be grown in embryonic brain cells from rabbits or rats with high virus yields. The cells became persistently infected and could be subcultivated without loss of infectivity. Cocultivation of infected rabbit brain (ERB) cells with GMK-, Vero-, or MDCK-cells led to persistently infected cell lines. BD virus grown in MDCK cells after cocultivation became adapted to this cell type and could be used directly for further infection of MDCK cells. PMID:6772932

Herzog, S; Rott, R

1980-01-01

117

Pro-angiogenic Cell Colonies Grown In Vitro from Human Peripheral Blood Mononuclear Cells  

PubMed Central

Although multiple culture assays have been designed to identify “endothelial progenitor cells” (EPCs), the phenotype of cells grown in culture often remains undefined. We sought to define and characterize the pro-angiogenic cell population within human peripheral blood mononuclear cells. Mononuclear cells were isolated from peripheral blood and grown under angiogenic conditions for 7 days. Formed colonies (CFU-As) were identified and analyzed for proliferation, mRNA and surface antigen expression, tube-forming ability and chromosomal content. Colonies were composed of a heterogeneous group of cells expressing the leukocyte antigens CD45, CD14, and CD3, as well as the endothelial proteins vascular endothelial (VE) cadherin, von Willebrand's Factor (vWF), CD31 and endothelial nitric oxide synthase (eNOS). Colony cells expressed increased levels of pro-angiogenic growth factors, and they formed tubes in Matrigel. In comparison with colonies from the CFU-Hill assay, our assay resulted in a greater number of colonies (19±9 vs. 13±7; p<0.0001) with a substantial number of cells expressing an endothelial phenotype (20.2±7.4% vs. 2.2±1.2% expressing eNOS, p=0006). Chromosomal analysis indicated the colony cells were bone marrow-derived. We, therefore, describe a colony forming unit assay that measures bone marrow-derived circulating mononuclear cells with the capacity to proliferate and mature into proangiogenic leukocytic and endothelial-like cells. This assay, therefore, reflects circulating, bone marrow-derived pro-angiogenic activity.

Mavromatis, Kreton; Sutcliffe, Diane; Joseph, Giji; Alexander, R. Wayne; Waller, Edmund K.; Quyyumi, Arshed A.; Taylor, W. Robert

2014-01-01

118

Single cell protein production by photosynthetic bacteria grown on the clarified effluents of biogas plant  

Microsoft Academic Search

Anaerobically digested cow dung was separated by centrifugation into solid residue and liquid supernatant fractions. Clarified supernatant fraction, rich in volatile fatty acids, supported the growth of photosynthetic bacteria. Single cell protein from different photosynthetic bacteria, grown on clarified supernatant, was found to be rich in essential and sulphur amino acids. Rhodopseudomonas capsulata produced the best single cell protein.

Sudhanshu Vrati; G. B. Pant

1984-01-01

119

Ultrastructure and enzyme complement of proplastids from heterotrophically grown cells of the red alga Galdieria sulphuraria  

Microsoft Academic Search

The unicellular thermoacidophilic red alga Galdieria sulphuraria is known to grow autotrophically as well as heterotrophically on a wide variety of organic carbon sources. Strain 107.79 becomes nearly colourless when grown heterotrophically on lactose. Under autotrophic conditions, a dominant chloroplast occupies most of the cell volume, while very small, yellow to colourless plastid-like organelles were observed in heterotrophic cells. Their

Gilbert Tischendorf; Christine Oesterhelt; Sabine Hoffmann; Jan Girnus; Claus Schnarrenberger; Wolfgang Gross

2007-01-01

120

Electron microscopic investigations of zymomonas mobilis cells grown in low and high glucose concentrations  

Microsoft Academic Search

Zymomonas mobilis cells grown in the presence of low and high glucose concentrations were examined by electron microscopy. Using ultrathin sectioning and agar diffusion method, significant changes in morphology were observed. Although the fine structure resembles that of a typical gram-negative bacterium, changes in glucose concentration and phases of growth lead to large cell wall vesicle or blebs formation. The

Horst W. Doelle; Hansdiirgen Preusser; Heidi Rostek

1982-01-01

121

Polar release of pathogenic Old World hantaviruses from renal tubular epithelial cells  

PubMed Central

Background Epithelio- and endotheliotropic viruses often exert polarized entry and release that may be responsible for viral spread and dissemination. Hantaviruses, mostly rodent-borne members of the Bunyaviridae family infect epithelial and endothelial cells of different organs leading to organ dysfunction or even failure. Endothelial and renal epithelial cells belong to the target cells of Old World hantavirus. Therefore, we examined the release of hantaviruses in several renal epithelial cell culture models. We used Vero cells that are commonly used in hantavirus studies and primary human renal epithelial cells (HREpC). In addition, we analyzed MDCKII cells, an epithelial cell line of a dog kidney, which represents a widely accepted in vitro model of polarized monolayers for their permissiveness for hantavirus infection. Results Vero C1008 and primary HREpCs were grown on porous-support filter inserts for polarization. Monolayers were infected with hantavirus Hantaan (HTNV) and Puumala (PUUV) virus. Supernatants from the apical and basolateral chamber of infected cells were analyzed for the presence of infectious particles by re-infection of Vero cells. Viral antigen and infectious particles of HTNV and PUUV were exclusively detected in supernatants collected from the apical chamber of infected Vero C1008 cells and HREpCs. MDCKII cells were permissive for hantavirus infection and polarized MDCKII cells released infectious hantaviral particles from the apical surface corresponding to the results of Vero and primary human epithelial cells. Conclusions Pathogenic Old World hantaviruses are released from the apical surface of different polarized renal epithelial cells. We characterized MDCKII cells as a suitable polarized cell culture model for hantavirus infection studies.

2012-01-01

122

Thin film solar cells grown by organic vapor phase deposition  

Microsoft Academic Search

Organic solar cells have the potential to provide low-cost photovoltaic devices as a clean and renewable energy resource. In this thesis, we focus on understanding the energy conversion process in organic solar cells, and improving the power conversion efficiencies via controlled growth of organic nanostructures. First, we explain the unique optical and electrical properties of organic materials used for photovoltaics,

Fan Yang

2008-01-01

123

Thin film solar cells grown by organic vapor phase deposition  

NASA Astrophysics Data System (ADS)

Organic solar cells have the potential to provide low-cost photovoltaic devices as a clean and renewable energy resource. In this thesis, we focus on understanding the energy conversion process in organic solar cells, and improving the power conversion efficiencies via controlled growth of organic nanostructures. First, we explain the unique optical and electrical properties of organic materials used for photovoltaics, and the excitonic energy conversion process in donor-acceptor heterojunction solar cells that place several limiting factors of their power conversion efficiency. Then, strategies for improving exciton diffusion and carrier collection are analyzed using dynamical Monte Carlo models for several nanostructure morphologies. Organic vapor phase deposition is used for controlling materials crystallization and film morphology. We improve the exciton diffusion efficiency while maintaining good carrier conduction in a bulk heterojunction solar cell. Further efficiency improvement is obtained in a novel nanocrystalline network structure with a thick absorbing layer, leading to the demonstration of an organic solar cell with 4.6% efficiency. In addition, solar cells using simultaneously active heterojunctions with broad spectral response are presented. We also analyze the efficiency limits of single and multiple junction organic solar cells, and discuss the challenges facing their practical implementations.

Yang, Fan

124

Assessment of MOCVD and MBE-grown GaAs for high-efficiency solar cell applications  

Microsoft Academic Search

A critical assessment of the photovoltaic quality of epitaxial GaAs grown by metal-organic chemical vapor deposition (MOCVD) and by molecular-beam epitaxy (MBE) is reported. Epitaxial films of nominally identical structure were grown by the two techniques and were fabricated into p-n heteroface solar cells. The 0.5-cm by 0.5-cm cells were then characterized and compared. The MOCVD-grown films produced independently verified

Stephen P. Tobin; S. M. Vernon; C. Bajgar; Steven J. Wojtczuk; Michael R. Melloch

1990-01-01

125

Commissioning and initial stereotactic ablative radiotherapy experience with Vero.  

PubMed

The purpose of this study is to describe the comprehensive commissioning process and initial clinical performance of the Vero linear accelerator, a new radiotherapy device recently installed at UT Southwestern Medical Center specifically developed for delivery of image-guided stereotactic ablative radiotherapy (SABR). The Vero system utilizes a ring gantry to integrate a beam delivery platform with image guidance systems. The ring is capable of rotating ± 60° about the vertical axis to facilitate noncoplanar beam arrangements ideal for SABR delivery. The beam delivery platform consists of a 6 MV C-band linac with a 60 leaf MLC projecting a maximum field size of 15 × 15 cm² at isocenter. The Vero planning and delivery systems support a range of treatment techniques, including fixed beam conformal, dynamic conformal arcs, fixed gantry IMRT in either SMLC (step-and-shoot) or DMLC (dynamic) delivery, and hybrid arcs, which combines dynamic conformal arcs and fixed beam IMRT delivery. The accelerator and treatment head are mounted on a gimbal mechanism that allows the linac and MLC to pivot in two dimensions for tumor tracking. Two orthogonal kV imaging subsystems built into the ring facilitate both stereoscopic and volumetric (CBCT) image guidance. The system is also equipped with an always-active electronic portal imaging device (EPID). We present our commissioning process and initial clinical experience focusing on SABR applications with the Vero, including: (1) beam data acquisition; (2) dosimetric commissioning of the treatment planning system, including evaluation of a Monte Carlo algorithm in a specially-designed anthropomorphic thorax phantom; (3) validation using the Radiological Physics Center thorax, head and neck (IMRT), and spine credentialing phantoms; (4) end-to-end evaluation of IGRT localization accuracy; (5) ongoing system performance, including isocenter stability; and (6) clinical SABR applications. PMID:24710458

Solberg, Timothy D; Medin, Paul M; Ramirez, Ezequiel; Ding, Chuxiong; Foster, Ryan D; Yordy, John

2014-01-01

126

Influence of Cell Age on Chlorophyll Formation in Light-grown and Etiolated Wheat Seedlings 1  

PubMed Central

A method is described for relating the age of a cereal leaf cell to its distance from the leaf base. The rates of chlorophyll synthesis per plastid in the first leaf of light-grown and of greening etiolated seedlings of wheat (Triticum aestivum, var. Maris Dove) increase with cell age. Normally developing plastids of light-grown wheat take over 24 hours to reach the chlorophyll a/b ratio characteristic of mature wheat chloroplasts (4.5), but mature etioplasts need only 8 hours light to achieve this a/b ratio. Plastid greening potential depends only on cell age, whereas the chlorophyll a/b ratio is influenced both by cell age and by light.

Boffey, Stephen A.; Sellden, Gun; Leech, Rachel M.

1980-01-01

127

Potassium activities in cell compartments of salt-grown barley leaves  

Microsoft Academic Search

Triple-barrelled microelectrodes measuring K+ activity (aK), pH and membrane potential were used to make quantitative measurements of vacuolar and cytosolic aK in epidermal and mesophyll cells of barley plants grown in nutrient solution with 0 or 200 mM added NaCl. Measurements of aK were assigned to the cytosol or vacuole based on the pH measured. In epidermal cells, the salt

Tracey Ann Cuin; Anthony J. Miller; Sophie A. Laurie; Roger A. Leigh

2003-01-01

128

Pd-cell purified hydrogen for highest purity AlGaAs grown by MOVPE  

Microsoft Academic Search

While the semiconductor industry's purity debate continues, we now know one thing for certain-carrier gas hydrogen from Pd-cell purifiers is purer than that made by getter-type purifiers according to DLTS measurements made in studies of AlGaAs grown by MOVPE, one of the industry's most demanding manufacturing processes. And this research shows that Pd-cell purified H2 also contributes to devices with

Wilson Chu; Johnson Matthey; Tom Purcell

1996-01-01

129

Structural and elemental characterization of heart cells grown in a collagen matrix  

Microsoft Academic Search

A novel preparation of spontaneously contracting heart cells embedded in a collagen strand provides an ideal experimental system for correlative structure-function experiments that utilize the techniques of electron microscopy, quantitative electron probe x-ray microanalysis (EPXMA) and imaging. Heart cells grown within the strand for 1 day possess the subcellular content and distribution of physiologically relevant elements--Na, Mg, P, S, Cl,

A. LeFurgey; L. A. Hawkey; P. Ingram; M. Lieberman

1991-01-01

130

Detailed Structural and Quantitative Analysis Reveals the Spatial Organization of the Cell Walls of in Vivo Grown Mycobacterium leprae and in Vitro Grown Mycobacterium tuberculosis*  

PubMed Central

The cell wall of mycobacteria consists of an outer membrane, analogous to that of Gram-negative bacteria, attached to the peptidoglycan (PG) via a connecting polysaccharide arabinogalactan (AG). Although the primary structure of these components is fairly well deciphered, issues such as the coverage of the PG layer by covalently attached mycolates in the outer membrane and the spatial details of the mycolic acid attachment to the arabinan have remained unknown. It is also not understood how these components work together to lead to the classical acid-fast staining of mycobacteria. Because the majority of Mycobacterium tuberculosis bacteria in established experimental animal infections are acid-fast negative, clearly cell wall changes are occurring. To address both the spatial properties of mycobacterial cell walls and to begin to study the differences between bacteria grown in animals and cultures, the cell walls of Mycobacterium leprae grown in armadillos was characterized and compared with that of M. tuberculosis grown in culture. Most fundamentally, it was determined that the cell wall of M. leprae contained significantly more mycolic acids attached to PG than that of in vitro grown M. tuberculosis (mycolate:PG ratios of 21:10 versus 16:10, respectively). In keeping with this difference, more arabinogalactan (AG) molecules, linking the mycolic acids to PG, were found. Differences in the structures of the AG were also found; the AG of M. leprae is smaller than that of M. tuberculosis, although the same basic structural motifs are retained.

Bhamidi, Suresh; Scherman, Michael S.; Jones, Victoria; Crick, Dean C.; Belisle, John T.; Brennan, Patrick J.; McNeil, Michael R.

2011-01-01

131

Co-culture Models of Pseudomonas aeruginosa Biofilms Grown on Live Human Airway Cells  

PubMed Central

Bacterial biofilms have been associated with a number of different human diseases, but biofilm development has generally been studied on non-living surfaces. In this paper, we describe protocols for forming Pseudomonas aeruginosa biofilms on human airway epithelial cells (CFBE cells) grown in culture. In the first method (termed the Static Co-culture Biofilm Model), P. aeruginosa is incubated with CFBE cells grown as confluent monolayers on standard tissue culture plates. Although the bacterium is quite toxic to epithelial cells, the addition of arginine delays the destruction of the monolayer long enough for biofilms to form on the CFBE cells. The second method (termed the Flow Cell Co-culture Biofilm Model), involves adaptation of a biofilm flow cell apparatus, which is often used in biofilm research, to accommodate a glass coverslip supporting a confluent monolayer of CFBE cells. This monolayer is inoculated with P. aeruginosa and a peristaltic pump then flows fresh medium across the cells. In both systems, bacterial biofilms form within 6-8 hours after inoculation. Visualization of the biofilm is enhanced by the use of P. aeruginosa strains constitutively expressing green fluorescent protein (GFP). The Static and Flow Cell Co-culture Biofilm assays are model systems for early P. aeruginosa infection of the Cystic Fibrosis (CF) lung, and these techniques allow different aspects of P. aeruginosa biofilm formation and virulence to be studied, including biofilm cytotoxicity, measurement of biofilm CFU, and staining and visualizing the biofilm.

Moreau-Marquis, Sophie; Redelman, Carly V.; Stanton, Bruce A.; Anderson, Gregory G.

2010-01-01

132

Co-culture models of Pseudomonas aeruginosa biofilms grown on live human airway cells.  

PubMed

Bacterial biofilms have been associated with a number of different human diseases, but biofilm development has generally been studied on non-living surfaces. In this paper, we describe protocols for forming Pseudomonas aeruginosa biofilms on human airway epithelial cells (CFBE cells) grown in culture. In the first method (termed the Static Co-culture Biofilm Model), P. aeruginosa is incubated with CFBE cells grown as confluent monolayers on standard tissue culture plates. Although the bacterium is quite toxic to epithelial cells, the addition of arginine delays the destruction of the monolayer long enough for biofilms to form on the CFBE cells. The second method (termed the Flow Cell Co-culture Biofilm Model), involves adaptation of a biofilm flow cell apparatus, which is often used in biofilm research, to accommodate a glass coverslip supporting a confluent monolayer of CFBE cells. This monolayer is inoculated with P. aeruginosa and a peristaltic pump then flows fresh medium across the cells. In both systems, bacterial biofilms form within 6-8 hours after inoculation. Visualization of the biofilm is enhanced by the use of P. aeruginosa strains constitutively expressing green fluorescent protein (GFP). The Static and Flow Cell Co-culture Biofilm assays are model systems for early P. aeruginosa infection of the Cystic Fibrosis (CF) lung, and these techniques allow different aspects of P. aeruginosa biofilm formation and virulence to be studied, including biofilm cytotoxicity, measurement of biofilm CFU, and staining and visualizing the biofilm. PMID:20972407

Moreau-Marquis, Sophie; Redelman, Carly V; Stanton, Bruce A; Anderson, Gregory G

2010-01-01

133

Efficacy and safety of cell-associated vaccines against Marek's disease virus grown in QT35 cells or JBJ-1 cells.  

PubMed

The Marek's disease virus (MDV) vaccine strain CVI 988 usually is grown in primary chicken embryo fibroblasts (CEFs). We found that the strains could be grown also in the QT35 and JBJ-1 cell lines to titers in the same range as in the CEFs. Both cell lines are fibroblast-like cell lines, which can be grown in flat-bottomed tissue-culture flasks, roller bottles, and on microcarriers. For growth in QT35 cells it was necessary to adapt the virus to the cell line; for growth in JBJ-1 cells this was not necessary. We investigated the efficacy of experimental CVI 988 vaccines grown in QT35 cells and JBJ-1 cells. The efficacy studies were performed in accordance with European Pharmacopoeia (EP) monograph for live MDV disease vaccines. Groups of 1-day-old specific-pathogen-free chicks were vaccinated. Nonvaccinated control groups were included in the studies. Five to 7 days after vaccination all chickens were challenged with the very virulent MDV strain RB1B. After challenge the chickens were observed for a period of 70 days for signs of MD. The protection induced by CVI 988 grown in QT35 cells as well as JBJ-1 cells complied with the requirements of the EP that prescribe that the protection index should be at least 80%. The safety of the vaccines grown in QT35 cells and JBJ-1 cells was tested in a field study in commercial layer chickens. The vaccine virus was not safe after passaging in QT35 cells. This can be explained by the presence of fragments of the genome of MDV strains in the QT35 cell line. No signs of MD were noticed in the study in which CVI988 grown in JBJ-1 cells was tested. It is concluded that the JBJ-1 cell line is a suitable substrate for the current vaccines against MD. PMID:23901760

Geerligs, Harm; Spijkers, Ine; Rodenberg, Jeff

2013-06-01

134

Efficient in situ electroporation of mammalian cells grown on microporous membranes.  

PubMed Central

Electroporation is a common technique for the introduction of DNA molecules into living cells. The method is currently limited by the necessity of applying the electrical discharge to cells in suspension. Adherent cells must therefore be removed from their substratum, which can induce unwanted physiological effects. We report here a new procedure for in situ electroporation of cells grown on microporous membranes of polyethylene terephthalate (PET) or polyester (PE). We demonstrate that this method of in situ electroporation employs only readily available materials and standard electroporation devices without any modifications, is as efficient as conventional electroporation of cells in suspension, and is applicable to a wide range of cell types. Efficient electroporation can be achieved under conditions of minimal cell killing, and can be performed with quiescent cells as well as with confluent epithelial sheets. The method is a useful extension of electroporation technology, and will allow the application of electroporation to a wider spectrum of biological systems. Images

Yang, T A; Heiser, W C; Sedivy, J M

1995-01-01

135

Effects of method of detachment on electrophoretic mobility of mammalian cells grown in monolayer culture  

NASA Technical Reports Server (NTRS)

A variety of proteolytic and micolytic enzumes, mechanical procedures, and changes in the ionic environment, especially Ca chelation, are used for dispersal of monolayer grown cells. If either chelating agents or mechanical dispersion are used alone, the cell yield is often low and suspensions of single cells are difficult to obtain. Confluent monolayers treated with EDTA tend to be released from their surfaces in sheets, and clumps of cells remain even after further incubation in EDTA. Crude trypsin is the most popular dispersal agent and is known to contain a variety of contaminating enzymes which contribute to the dispersal of cells. A variety of cell injuries resulting from the activity of proteolytic enzymes are reported. It is shown that crystalline trypsin is least harmful to cell integrity as judged by trypan blue uptake.

Plank, L. D.; Kunze, M. E.; Todd, P. W.

1985-01-01

136

Molecular beam epitaxy grown GaNAsSb 1 eV photovoltaic cell  

Microsoft Academic Search

We report the performance of a 1 eV GaNAsSb-based photovoltaic cell grown using a molecular beam epitaxy system equipped with a radio frequency (RF) plasma-assisted nitrogen source. The 1 mum-thick photoabsorption layer contains 2% of N and 6% of Sb resulting in a GaNAsSb layer with bandgap energy of 1.0 eV. Under AM1.5G solar illumination condition with and without 850

K. H. Tan; S. Wicaksono; W. K. Loke; D. Li; S. F. Yoon; E. A. Fitzgerald; S. A. Ringel; J. S. Harris Jr.

2011-01-01

137

Tilted bulk heterojunction organic photovoltaic cells grown by oblique angle deposition  

Microsoft Academic Search

We demonstrate small molecule bulk heterojunction organic photovoltaic cells using oblique angle vacuum deposition. Obliquely deposited donor chloroaluminum phthalocyanine (ClAlPc) films on indium tin oxide have surface feature sizes of ~30 nm, resulting in ClAlPc\\/C60 donor-acceptor heterojunctions (HJs) with approximately twice the interface area of HJs grown at normal incidence. This results in nearly twice the external quantum efficiency in

Ning Li; Stephen R. Forrest

2009-01-01

138

Response to Ionizing Radiation of Human Bladder Transitional Cell Carcinomas Grown in the Nude Mouse  

Microsoft Academic Search

The sensitivity to ionizing radiation of three human bladder transitional cell carcinomas grown in the nude mouse was investigated at four different radiation levels, 500, 1,000, 2,000 and 4,000 rad. Each tumor line exhibited a response pattern which reflected to a certain degree tumor differentiation and growth rate in the nude mouse. Exposure to 1,000 rad was the minimum amount

Andreas P. Kyriazis; Alan Yagoda; Aikaterini A. Kyriazis; James G. Kereiakes; William B. McCombs; III

1985-01-01

139

Lithium Induces ER Stress and N-Glycan Modification in Galactose-Grown Jurkat Cells  

PubMed Central

We previously reported that lithium had a significant impact on Ca2+ regulation and induced unfolded protein response (UPR) in yeast cells grown on galactose due to inhibition of phosphoglucomutase (PGM), however the exact mechanism has not been established yet. In this study, we analysed lithium's effect in galactose-fed cells to clarify whether these ER-related changes are the result of a relative hypoglycemic state. Furthermore, we investigated whether the alterations in galactose metabolism impact protein post-translational modifications. Thus, Jurkat cells were incubated in glucose or galactose containing media with or without lithium treatment. We found that galactose-fed and lithium treated cells showed better survivability than fasting cells. We also found higher UDP-Hexose and glycogen levels in these cells compared to fasting cells. On the other hand, the UPR (X-box binding protein 1 mRNA levels) of galactose-fed and lithium treated cells was even greater than in fasting cells. We also found increased amount of proteins that contained N-linked N-acetyl-glucosamine, similar to what was reported in fasting cells by a recent study. Our results demonstrate that lithium treatment of galactose-fed cells can induce stress responses similar to hypoglycemia, however cell survival is still secured by alternative pathways. We propose that clarifying this process might be an important addition toward the better understanding of the molecular mechanisms that regulate ER-associated stress response.

Katai, Emese; Yahiro, Rikki K. K.; Poor, Viktor S.; Montsko, Gergely; Zrinyi, Zita; Kovacs, Gabor L.; Miseta, Attila

2013-01-01

140

Structural and elemental characterization of heart cells grown in a collagen matrix  

SciTech Connect

A novel preparation of spontaneously contracting heart cells embedded in a collagen strand provides an ideal experimental system for correlative structure-function experiments that utilize the techniques of electron microscopy, quantitative electron probe x-ray microanalysis (EPXMA) and imaging. Heart cells grown within the strand for 1 day possess the subcellular content and distribution of physiologically relevant elements--Na, Mg, P, S, Cl, K, and Ca--found in intact heart cell preparations. The presence of junctional specializations between, and organized myofibrils within, the majority of cells after 1 day in culture also establishes that the collagen matrix promotes vigorous cell development as well as maintains physiological integrity. EPXMA, combined with ultrastructural analyses, provides elemental content data on a cell-by-cell basis. In studies presented here, viable cells, comprising over 80% of the strand cell population, could be distinguished easily from those which had been functionally compromised, not only by aberrant structure but also by altered subcellular compartmentation of Na, K, Cl, and Ca. Within individual viable cells, compartmental differences in element content were notable especially between mitochondria and cytoplasm. However, nuclear euchromatin, but not heterochromatin, appeared approximately identical to cytoplasm in elemental and water content. In such cells, the cytoplasmic K:Na ratio was maintained at a high level (approximately 15:1). The results with respect to K, Na, and other elements demonstrated the integrity of membrane transport mechanisms regulating the movement and distribution of ions and the maintenance of ionic homeostasis in cells of the strand preparation.

LeFurgey, A.; Hawkey, L.A.; Ingram, P.; Lieberman, M. (Duke Univ. Medical Center, Durham, NC (USA))

1991-02-01

141

Evaluation of different yeast cell wall mutants and microalgae strains as feed for gnotobiotically grown brine shrimp Artemia franciscana  

Microsoft Academic Search

The nutritional value of isogenic yeast strains and two microalgal species for gnotobiotically grown Artemia was examined. Yeast cell wall mutants were always better feed for Artemia than their respective wild type. Yeast cells harbouring null mutants for enzymes involved early in the biochemical pathway for cell wall mannoproteins synthesis performed best as feed for Artemia. Yeast cells defective in

Antonio Marques; Jean Dhont; Patrick Sorgeloos; Peter Bossier

2004-01-01

142

Cyclic-radiation response of murine fibrosarcoma cells grown as pulmonary nodules  

SciTech Connect

The radiation age response of murine fibrosarcoma (FSa) cells grown as pulmonary nodules in C/sub 3/Hf/Kam mice was determined. FSa cells were irradiated in vivo either with 10 Gy as 14 day-old lung tumors (i.e., artificial macrometastases) prior to cell separation or with 5 Gy as single cells trapped in the lungs of recipient mice (i.e., artificial micrometastases) following cell separation and synchronization by centrifugal elutriation. Flow microfluorometry (FMF) was used to determine cell-cycle parameters and the relative synchrony of the separated populations, as well as the percent contamination of normal diploid cells in each of the tumor cell populations. Tumor populations containing up to 90% G/sub 1/, 60% S-, and 75% G/sub 2/+M-phase tumor cells were obtained. Cell clonogenicity, determined using a lung colony assay, ranged from 0.7 to 6% for control FSa cells from the various elutriator fractions. The radiation sensitivity of these separated cell populations varied by a factor of 6, regardless of whether the cells were irradiated as artificial micro or macro-metastases. In each experiment, tumor populations most enriched in s-phase cells exhibited the greatest radiation sensitivity. To confirm that these populations were highly enriched in S-phase cells and to demonstrate that they were more radiosensitive than FSa cells in other parts of the cell cycle, the elutriated tumor populations were exposed to either suicide labeling by high specific activity tritiated thymidine or hydroxyurea. The resultant age response curves were qualitatively similar to those obtained following irradiation and reflected the S-phase sensitivity of FSa cells to these agents.

Grdina, D.J.; Hunter, N.

1982-10-01

143

Glucose uptake rates of single E. coli cells grown in glucose-limited chemostat cultures.  

PubMed

To evaluate the extent to which single-cell glucose uptake rates determine the overall specific growth rate of a culture, dilute chemostat cultures of Escherichia coli BL21 were grown in defined medium under glucose limitation. The glucose uptake dynamics of the cell population was examined at the single-cell level using the fluorescent glucose analog, 2-NBDG. Between dilution rates of 0.12 h(-1) and 0.40 h(-1), mean cellular protein content and steady-state, extracellular glucose concentrations increased with increasing dilution rate. However, the distribution of 2-NBDG uptake rates in the population remained constant over the range of dilution rates studied. This indicates that the growth of cells in continuous culture is not limited by the maximum rate of uptake of glucose but by the availability of glucose for transport. The work represents an example of how quantitative flow cytometry can be applied to gain detailed insight into microbial growth physiology. PMID:11000435

Natarajan, A; Srienc, F

2000-09-01

144

Cyclic-radiation response of murine fibrosarcoma cells grown as pulmonary nodules  

SciTech Connect

The radiation age response of murine fibrosarcoma (FSa) cells grown as pulmonary nudules in C/sub 3/Hf/Kam mice was determined. FSa cells were irradiated in vivo either with 10 Gy as 14 day-old lung tumors (i.e., artifical micrometastases) following cell separation and synchronization by centrifugal elutriation. Flow microfluorometry (FMF) was used to determine cell-cycle parameters and the relative synchrony of the separated populations, as well as the percent contamination of normal diploid cells in each of the tumor cells populations. Tumor populations containing up to 90% G/sub 1/-, 60% S-, and 75% G/sub 2/+M-phase tumor cells were obtained. Cell clonogenicity, determined using a lung colony assay, ranged from 0.7 to 6% for control FSa cells from the various elutriator fractions. The radiation sensitivity of these separated cell populations varied by a factor of 6, regardless of whether the cells were irradiated as artifical micro or macro-metastases. In each experiment, tumor population most enriched in S-phase cells exhibited the greatest radiation sensitivity. To confirm that these populations were highly enriched in S-phase cells and to demonstrate that they were more radiosensitive than FSa cells in other parts of the cell cycle, the elutriated tumor population were exposed to either suicide labeling by high specific activity tritated thymidine or hydroxyurea. The resultant age response curves were qualitatively similar to those obtained following irradiation and reflected the S-phase sensitivity of FSa cells to these agents.

Grdina, D.J.; Hunter, N.

1982-10-01

145

Molecular beam epitaxy grown GaNAsSb 1 eV photovoltaic cell  

Microsoft Academic Search

We report the performance of a 1eV GaNAsSb-based photovoltaic cell grown using a molecular beam epitaxy system equipped with a radio frequency (RF) plasma-assisted nitrogen source. The 1?m-thick photoabsorption layer contains 2% of N and 6% of Sb resulting in a GaNAsSb layer with bandgap energy of 1.0eV. Under AM1.5G solar illumination condition with and without 850nm long pass filter,

K. H. Tan; S. Wicaksono; W. K. Loke; D. Li; S. F. Yoon; E. A. Fitzgerald; S. A. Ringel; J. S. Harris Jr.

2011-01-01

146

Complementary Distribution of NN and NNO Complexes in Cast-Grown Multicrystalline Silicon for Photovoltaic Cells  

NASA Astrophysics Data System (ADS)

Multicrystalline silicon grown by the cast method and used in photovoltaic cells includes nitrogen from the Si3N4 coating of the SiO2 crucible. We investigated the distribution of nitrogen, existing as NN or NNO complexes in silicon, by Fourier transform infrared spectroscopy. We found that the NN peak intensity was higher at the upper part of the silicon ingot, while the NNO peak intensity was higher at the lower part. There was a complementary relationship between the distribution of NN and that of NNO; this implied that NNO complexes were formed by NN complexes that had combined with interstitial oxygen during crystal growth.

Kusunoki, Hiroki; Ishizuka, Takahide; Ogura, Atsushi; Ono, Haruhiko

2011-11-01

147

Molecular beam epitaxy grown GaNAsSb 1 eV photovoltaic cell  

NASA Astrophysics Data System (ADS)

We report the performance of a 1 eV GaNAsSb-based photovoltaic cell grown using a molecular beam epitaxy system equipped with a radio frequency (RF) plasma-assisted nitrogen source. The 1 ?m-thick photoabsorption layer contains 2% of N and 6% of Sb resulting in a GaNAsSb layer with bandgap energy of 1.0 eV. Under AM1.5G solar illumination condition with and without 850 nm long pass filter, the GaNAsSb-based photovoltaic cell demonstrates a JSC values of 15 and 32 mA/W, respectively. Deep level transient spectroscopy analysis reveals that the VOC of the photovoltaic cell could possibly be limited by the presence of arsenic antisite defects.

Tan, K. H.; Wicaksono, S.; Loke, W. K.; Li, D.; Yoon, S. F.; Fitzgerald, E. A.; Ringel, S. A.; Harris, J. S., Jr.

2011-11-01

148

LiNixCo1-xO2 Cell Grown by Pulsed Laser Deposition  

NASA Astrophysics Data System (ADS)

Thin films of LiNixCo1-xO2 were prepared by pulsed laser deposition technique. Two important deposition parameters such as substrate temperature and oxygen partial pressure during the thin film deposition were controlled. The electrochemical measurements were carried out on Li//LiNixCo1-xO2 cells with a lithium metal foil as anode and LiNixCo1-xO2 film as cathode of 1.5 cm2 active area using a Teflon home-made cell hardware. Electrochemical titration was made by charging and discharging the cells using the galvanostatic mode of a Mac-Pile single 608 electrochemical analyzer system in the potential range between 2.0 and 4.1 V. Specific capacity as high as 220 mC/cm2 ?m was measured for the film grown at 700 °C.

Rao, M. C.; Ravindranadh, K.; Begum, Sk. Muntaz; Nirmala, G.

2011-07-01

149

Modification of cell wall architecture of wheat coleoptiles grown under hypergravity conditions.  

PubMed

Cell wall structure of wheat coleoptiles grown under continuous hypergravity (300 g) conditions was investigated. Length of coleoptiles exposed to hypergravity for 2-4 days from germination stage was 60-70% of that of 1 g control. The amounts of cell wall polysaccharides substantially increased during the incubation period both in 1 g control and hypergravity-treated coleoptiles. As a results, the levels of cell wall polysaccharides per unit length of coleoptile, which mean the thickness of cell walls, largely increased under hypergravity conditions. The major sugar components of the hemicellulose fraction, a polymer fraction extracted from cell walls with strong alkali, were arabinose (Ara), xylose (Xyl) and glucose (Glc). The molar ratios of Ara and Xyl to Glc in hypergravity-treated coleoptiles were higher than those in control coleoptiles. Furthermore, the fractionation of hemicellulosic polymers into the neutral and acidic polymers by the anion-exchange column showed that the levels of acidic polymers in cell walls of hypergravity-treated coleoptiles were higher than those of control coleoptiles. These results suggest that hypergravity stimuli bias the synthesis of hemicellulosic polysaccharides and increase the proportion of acidic polymers, such as arabinoxylans, in cell walls of wheat coleoptiles. These structural changes in cell walls may contribute to plant resistance to hypergravity stimuli. PMID:14676390

Wakabayashi, Kazuyuki; Soga, Kouichi; Kamisaka, Seiichiro; Hoson, Takayuki

2003-10-01

150

Phase III clinical trials comparing the immunogenicity and safety of the vero cell-derived Japanese encephalitis vaccine Encevac with those of mouse brain-derived vaccine by using the Beijing-1 strain.  

PubMed

The immunogenicity and safety of an inactivated cell culture Japanese encephalitis vaccine (CC-JEV) were compared with those of an inactivated mouse brain-derived Japanese encephalitis vaccine (MB-JEV) in phase III clinical multicenter trials conducted in children. The vaccines contain the same Japanese encephalitis virus strain, the Beijing-1 strain. Two independent clinical trials (trials 1 and 2) were conducted. Trial 1 was conducted in 468 healthy children. Each subject was injected with 17 ?g per dose of either CC-JEV or MB-JEV, and the immunogenicity and safety of the vaccines were investigated. Trial 1 showed that CC-JEV was more immunogenic and reactive than MB-JEV at the same dose. Therefore, to adjust the immunogenicity of CC-JEV to that of MB-JEV, a vaccine that has had a good track record regarding its efficacy for a long time, trial 2 was conducted in 484 healthy children. To improve the stability, CC-JEV was converted from a liquid type to a freeze-dried type of vaccine. Each subject was injected subcutaneously with either 4 ?g per dose of CC-JEV, 8 ?g per dose of CC-JEV, or 17 ?g per dose of MB-JEV twice, at an interval of 2 to 4 weeks, followed by an additional booster immunization 1 to 15 months after the primary immunization. Based on the results of trial 2, 4 ?g per dose of the freeze-dried CC-JEV (under the label Encevac) was selected as a substitute for the MB-JEV. Encevac was approved and launched in 2011 and has since been in use as a 2nd-generation Japanese encephalitis vaccine in Japan. (These studies have been registered at the JapicCTI under registration no. JapicCTI-132063 and JapicCTI-080586 for trials 1 and 2, respectively). PMID:24334689

Miyazaki, Chiaki; Okada, Kenji; Ozaki, Takao; Hirose, Mizuo; Iribe, Kaneshige; Yokote, Hiroyuki; Ishikawa, Yuji; Togashi, Takehiro; Ueda, Kohji

2014-02-01

151

Distribution of Cytochrome c Peroxidase Activity in Wild-Type and Petite Cells of Bakers' Yeast Grown Aerobically and Anaerobically  

PubMed Central

Studies of mitochondrial biogenesis in yeast have been hampered by a lack of suitable membrane markers in anaerobically grown cells subsequently grown in air. Cytochrome c peroxidase activity and subcellular location was studied to determine whether it would be a useful marker for an analysis of mitochondrial formation. Cytochemical tests revealed enzyme reaction product on all mitochondrial membranes in aerobically grown wild-type cells. Anaerobically grown wild-type and all petite cultures contained cytochrome c peroxidase cytochemical reaction deposits on abundant cytoplasmic membranes and on the few mitochondrial profiles which also were seen in the electron photomicrographs. Biochemical studies corroborated the cytochemistry because mitochondrial fractions were greatly enriched in cytochrome c peroxidase activity for aerobically grown wild-type cultures, but petite and anaerobically grown wild-type cultures showed higher enzyme activities in supernatant fractions than was present in the corresponding particulate fractions after differential centrifugation. Evidence from low-temperature microspectroscopy, spectrophotometric assays of mitochondrial enzyme activities, and electron microscopy showed mitochondrial formation during the time required for preparation and lysis of spheroplasts from anaerobically grown cultures. The data were interpreted as indicating that cytochrome c peroxidase was an oxygen-inducible enzyme, and that there was a developmental relationship between enzyme-reactive membranes of mitochondria and cytoplasm during the period of respiratory adaptation. Images

Avers, Charlotte J.

1967-01-01

152

Structural and elemental characterization of heart cells grown in a collagen matrix.  

PubMed

A novel preparation of spontaneously contracting heart cells embedded in a collagen strand provides an ideal experimental system for correlative structure-function experiments that utilize the techniques of electron microscopy, quantitative electron probe x-ray microanalysis (EPXMA) and imaging. Heart cells grown within the strand for 1 day possess the subcellular content and distribution of physiologically relevant elements--Na, Mg, P, S, Cl, K, and Ca--found in intact heart cell preparations. The presence of junctional specializations between, and organized myofibrils within, the majority of cells after 1 day in culture also establishes that the collagen matrix promotes vigorous cell development as well as maintains physiological integrity. EPXMA, combined with ultrastructural analyses, provides elemental content data on a cell-by-cell basis. In studies presented here, viable cells, comprising over 80% of the strand cell population, could be distinguished easily from those which had been functionally compromised, not only by aberrant structure but also by altered subcellular compartmentation of Na, K, Cl, and Ca. Within individual viable cells, compartmental differences in element content were notable especially between mitochondria and cytoplasm. However, nuclear euchromatin, but not heterochromatin, appeared approximately identical to cytoplasm in elemental and water content. In such cells, the cytoplasmic K:Na ratio was maintained at a high level (approximately 15:1). The results with respect to K, Na, and other elements demonstrated the integrity of membrane transport mechanisms regulating the movement and distribution of ions and the maintenance of ionic homeostasis in cells of the strand preparation. PMID:2059550

LeFurgey, A; Hawkey, L A; Ingram, P; Lieberman, M

1991-02-01

153

Human norovirus infection of caco-2 cells grown as a three-dimensional tissue structure.  

PubMed

Human norovirus (hNoV) infectivity was studied using a three-dimensional model of large intestinal epithelium. Large intestine Caco-2 cells were grown in rotating wall vessel bioreactors for 18-21 days at 37 degrees C and then transferred to 24-well tissue culture plates where they were infected with GI.1 and GII.4 human noroviruses collected from human challenge trials and various outbreak settings, respectively. Compared with uninfected cells, transmission micrographs of norovirus-infected cells displayed evidence of shortening or total loss of apical microvilli, and vacuolization. Quantitative reverse transcription real-time PCR (qRT-PCR) indicated an approximate 2-3 log10 increase in viral RNA copies for the infected cells. A passage experiment examined both the ability for continued viral RNA and viral antigen detection. In the passaged samples 1.01x10(6) copies ml(-1) were detected by qRT-PCR. Immune electron microscopy using primary antibody to hNoV GI.1 capsids in conjunction with 6 nm gold-labelled secondary antibodies was performed on crude cellular lysates. Localization of antibody was observed in infected but not for uninfected cells. Our present findings, coupled with earlier work with the three-dimensional small intestinal INT407 model, demonstrate the utility of 3-D cell culture methods to develop infectivity assays for enteric viruses that do not readily infect mammalian cell cultures. PMID:21942189

Straub, Timothy M; Bartholomew, Rachel A; Valdez, Catherine O; Valentine, Nancy B; Dohnalkova, Alice; Ozanich, Richard M; Bruckner-Lea, Cynthia J; Call, Douglas R

2011-06-01

154

Human Norovirus Infection of Caco-2 Cells Grown as a 3-Dimensional Tissue Structure  

PubMed Central

Human norovirus (hNoV) infectivity was studied using a 3-dimensional model of large intestinal epithelium. Large intestine Caco-2 cells were grown in rotating wall vessel bioreactors for 18-21 days at 37°C and then transferred to 24-well tissue culture plates where they were infected with GI.1 and GII.4 human noroviruses collected from human challenge trials and various outbreak settings, respectively. Compared to uninfected cells, transmission micrographs of norovirus infected cells displayed evidence of shortening or total loss of apical microvilli, and vacuolization. Quantitative reverse transcription real-time PCR (qRT-PCR) indicated an approximate 2-3 Log10 increase in viral RNA copies for the infected cells. A passage experiment examined both the ability for continued viral RNA and viral antigen detection. In the passaged samples 1.01 × 106 copies/mL were detected by qRT-PCR. Immune electron microscopy using primary antibody to hNoV GI.1 capsids in conjunction with 6 nm gold-labeled secondary antibodies was performed on crude cellular lysates. Localization of antibody was observed in infected but not for uninfected cells. Our present findings, coupled with earlier work with the 3-dimensional small intestinal INT407 model, demonstrate the utility of 3-D cell culture methods to develop infectivity assays for enteric viruses that do not readily infect mammalian cell cultures.

Bartholomew, Rachel A.; Valdez, Catherine O.; Valentine, Nancy B.; Dohnalkova, Alice; Ozanich, Richard M.; Bruckner-Lea, Cynthia J.; Call, Douglas R.

2011-01-01

155

Comparison of Chloroflexus aurantiacus strain J-10-fl proteomes of cells grown chemoheterotrophically and photoheterotrophically  

SciTech Connect

Chloroflexus aurantiacus J-10-fl is a thermophilic green bacterium, a filamentous anoxygenic phototroph, and the model organism of the phylum Chloroflexi. We applied high-throughput, liquid chromatography-mass spectrometry in a global quantitative proteomics investigation of C. aurantiacus cells grown under oxic (chemoorganoheterotrophically) and anoxic (photoorganoheterotrophically) redox states. Our global analysis identified 13,524 high-confidence peptides that matched to 1,286 annotated proteins, 242 of which were either uniquely identified or significantly increased in abundance under anoxic culture conditions. Fifty-three of the 242 proteins are previously characterized photosynthesis-related proteins, including chlorosome proteins, proteins involved in the bacteriochlorophyll biosynthesis, 3-hydroxypropionate (3-OHP) CO2 fixation pathway, and components of electron transport chains. The remaining 190 proteins have not previously been reported. Of these, five proteins were found to be encoded by genes from a novel operon and observed only in photoheterotrophically grown cells. These proteins candidates may prove useful in further deciphering the phototrophic physiology of C. aurantiacus and other filamentous anoxygenic phototrophs.

Cao, Li; Bryant, Donald A.; Schepmoes, Athena A.; Vogl, Kajetan; Smith, Richard D.; Lipton, Mary S.; Callister, Stephen J.

2012-01-17

156

Enhanced chemoresistance of squamous carcinoma cells grown in 3D cryogenic electrospun scaffolds.  

PubMed

It is critically important to study head and neck squamous cell carcinoma tumorigenic mechanisms in order to gain a better understanding of tumor development, progression, and treatment. Unfortunately, a representative three-dimensional (3D) model for these evaluations has yet to be developed. The purpose of this study was to replicate tumor extracellular matrix (ECM) morphology utilizing electrospinning technology. First, the tumor ECM was evaluated by decellularizing tumor samples and analyzing the fibrous structure of the ECM by scanning electron microscopy. Cryogenic electrospun silk scaffolds were then fabricated to mimic the tumor ECM, and were found to be similar in fiber orientation and fiber dimensions to the native tumor ECM. Tumor cells were cultured on these ECM mimicking scaffolds and compared to an in vivo model of the same derivative human tumor in terms of proliferation and differentiation. The tumor cells in the 3D model show similar phenotypes to those found in vivo, contrasting to the same cells grown in two-dimensional (2D) culture. The sensitivity of the tumor cells to paclitaxel was compared between 2D culture and 3D culture. The results indicate that increased drug concentrations, orders of magnitude higher than the IC90 for 2D culture, had minimal effects on HN12 cell viability in the 3D model. In conclusion, an in vitro tumor model has been developed that will allow for a better understanding of tumor biology and aid chemotherapeutic drug development and accurate evaluation of drug efficacy. PMID:24057893

Bulysheva, Anna A; Bowlin, Gary L; Petrova, Stella P; Yeudall, W Andrew

2013-10-01

157

Electron-cytochemical study of Ca2+ in cotyledon cells of soybean seedlings grown in microgravity  

NASA Technical Reports Server (NTRS)

Microgravity and horizontal clinorotation are known to cause the rearrangement of the structural-functional organization of plant cells, leading to accelerated aging. Altered gravity conditions resulted in an increase in the droplets volume in cells and the destruction of chloroplast structure in Arabidopsis thaliana plants, an enhancement of cytosolic autophagaous processes, an increase in the respiration rate and a greater number of multimolecular forms of succinate- and malate dehydrogenases in cells of the Funaria hygrometrica protonema and Chlorella vulgaris, and changes in calcium balance of cells. Because ethylene is known to be involved in cell aging and microgravity appears to speed the process, and because soybean seedlings grown in space produce higher ethylene levels we asked: 1) does an acceleration of soybean cotyledon cell development and aging occur in microgravity? 2) what roles do Ca2+ ions and the enhanced ethylene level play in these events? Therefore, the goal of our investigation was to examine of the interaction of microgravity and ethylene on the localization of Ca2+ in cotyledon mesophyll of soybean seedlings.

Nedukha, O.; Brown, C. S.; Kordyum, E.; Piastuch, W. C.; Guikema, J. A. (Principal Investigator)

1999-01-01

158

Subcellular fractionation and morphology of calf aortic smooth muscle cells: studies on whole aorta, aortic explants, and subcultures grown under  

PubMed Central

A comparative biochemical and morphological study was made of calf aortic smooth muscle cells found in situ and grown in vitro under various conditions. Striking alterations in enzyme contents, physical properties, and morphological appearances of lysosomes, endoplasmic reticulum, plasma membranes and, to a lesser extent, mitochondria were observed upon culturing of calf aortic smooth muscle cells. These changes first appeared in cells growing out of tissue explants. They developed further upon subculturing of the cells and depended greatly on the culture conditions used. The alterations included increases in specific activities of some 5- to 25-fold of four acid hydrolases, an average ninefold increase in 5' -nucleotidase, sevenfold increase in cytochrome oxidase, and fourfold increase in neutral ?-glucosidase in subcultured smooth muscle cells compared to aortic cells in situ. Cell fractionation studies showed significant shifts in the equilibrium densities of plasma membranes, microsomes, and lysosomes, but not of mitochondria, in smooth muscle cells growing out from explants and in subcultured cells, compared to cells isolated from intact aortas. Although the cells grown in vitro exhibited typical phenotypic features of smooth muscle cells such as abundant myofilaments and surface vesicles, alterations in the morphological appearance of the endoplasmic reticulum, Golgi apparatus, and, especially, lysosomes were observed. These results demonstrate significant differences in specific cellular characteristics and functions of aortic smooth muscle cells grown in vitro compared to aortic cells in situ.

Fowler, S; Shio, H; Wolinsky, H

1977-01-01

159

Differential Response in Downstream Processing of CHO Cells Grown Under Mild Hypothermic Conditions  

PubMed Central

The manufacture of complex therapeutic proteins using mammalian cells is well established, with several strategies developed to improve productivity. The application of sustained mild hypothermic conditions during culture has been associated with increases in product titer and improved product quality. However, despite associated cell physiological effects, very few studies have investigated the impact on downstream processing (DSP). Characterization of cells grown under mild hypothermic conditions demonstrated that the stationary phase was prolonged by delaying the onset of apoptosis. This enabled cells to maintain viability for extended periods and increase volumetric productivity from 0.74 to 1.02 g L?1. However, host cell proteins, measured by ELISA, increased by ?50%, attributed to the extended time course and higher peak and harvest cell densities. The individual components making up this impurity, as determined by SELDI-TOF MS and 2D-PAGE, were shown to be largely comparable. Under mild hypothermic conditions, cells were less shear sensitive than those maintained at 37°C, enhancing the preliminary primary recovery step. Adaptive changes in membrane fluidity were further investigated by adopting a pronounced temperature shift immediately prior to primary recovery and the improvement observed suggests that such a strategy may be implementable when shear sensitivity is of concern. Early and late apoptotic cells were particularly susceptible to shear, at either temperature, even under the lowest shear rate investigated. These findings demonstrate the importance of considering the impact of cell culture strategies and cell physiology on DSP, by implementing a range of experimental methods for process characterization. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:688–696, 2013

Tait, Andrew S; Tarrant, Richard D R; Velez-Suberbie, M Lourdes; Spencer, Daniel I R; Bracewell, Daniel G

2013-01-01

160

Label-free optical detection of cells grown in 3D silicon microstructures.  

PubMed

We demonstrate high aspect-ratio photonic crystals that could serve as three-dimensional (3D) microincubators for cell culture and also provide label-free optical detection of the cells. The investigated microstructures, fabricated by electrochemical micromachining of standard silicon wafers, consist of periodic arrays of silicon walls separated by narrow deeply etched air-gaps (50 ?m high and 5 ?m wide) and feature the typical spectral properties of photonic crystals in the wavelength range 1.0-1.7 ?m: their spectral reflectivity is characterized by wavelength regions where reflectivity is high (photonic bandgaps), separated by narrow wavelength regions where reflectivity is very low. In this work, we show that the presence of cells, grown inside the gaps, strongly affects light propagation across the photonic crystal and, therefore, its spectral reflectivity. Exploiting a label-free optical detection method, based on a fiberoptic setup, we are able to probe the extension of cells adherent to the vertical silicon walls with a non-invasive direct testing. In particular, the intensity ratio at two wavelengths is the experimental parameter that can be well correlated to the cell spreading on the silicon wall inside the gaps. PMID:23817434

Merlo, Sabina; Carpignano, Francesca; Silva, Gloria; Aredia, Francesca; Scovassi, A Ivana; Mazzini, Giuliano; Surdo, Salvatore; Barillaro, Giuseppe

2013-08-21

161

Investigation of Indium Gallium Nitride Grown via Metal Organic Chemical Vapor Deposition in Various Crystallographic Orientations for Solar Cell Applications  

NASA Astrophysics Data System (ADS)

Solar cell technology has long relied upon Si and GaAs based materials. While this industry is mature, it has approached a plateau in the push to increase efficiency. It has been proposed that the InGaN ternary materials system is ideal for this purpose. In this work we report on the growth, fabrication and testing of photovoltaic properties of InGaN based solar cells grown via metalorganic chemical vapor deposition (MOCVD). In order for solar cells to work effectively, a minimum active region thickness is necessary for sufficient absorption of photons for conversion. At low In content compositions, high quality material has been grown and a simple p-i-n type solar cell with a single absorbing layer can be produced. However, at high In content compositions critical thickness limits for the thin films are well below the thickness requirements for full absorption. High In compositions are necessary to efficiently match the solar spectrum when designing multijunction solar cells. Solar cells require a thickness of 100-200 nm for sufficient absorption. Growth optimization and results for p-i-n type: single double heterostructures, multiple double heterostructure (MDH), and MQW, solar cells will be reported for InGaN grown on sapphire in the +c [0001] orientation as well as InGaN grown on bulk m-plane (10-10) substrates. Non-polar oriented growth of InGaN was investigated since as In content increases in typical c-plane growth, the polarization fields in the double heterostructure design of the p-i-n solar cell oppose the built in field of the junction and could potentially hinder carrier collection. At low In content, m-plane InGaN solar cells outperform c-plane solar cells with the same composition due to the higher quality material grown homoepitaxially on bulk GaN substrates.

Cruz, Samantha Christine

162

Electrochemically grown ZnO nanorods for hybrid solar cell applications  

SciTech Connect

A hybrid solar cell is designed and proposed as a feasible and reasonable alternative, according to acquired efficiency with the employment of zinc oxide (ZnO) nanorods and ZnO thin films at the same time. Both of these ZnO structures were grown electrochemically and poly(3-hexylthiophene):phenyl-C61-butyric acid methyl ester; (P3HT:PCBM) was used as an active polymer blend, which was found to be compatible to prepared indium-tin-oxide (ITO) substrate base. This ITO base was introduced with mentioned ZnO structure in such a way that, the most efficient configuration was optimized to be ITO/ZnO film/ZnO nanorod/P3HT: PCBM/Ag. Efficiency of this optimized device is found to be 2.44%. All ZnO works were carried out electrochemically, that is indeed for the first time and at relatively lower temperatures. (author)

Hames, Yakup [Department of Electrical-Electronics Engineering, Mustafa Kemal University, 31040 Hatay (Turkey); Alpaslan, Zuehal; Koesemen, Arif; San, Sait Eren; Yerli, Yusuf [Department of Physics, Gebze Institute of Technology, 41400 Gebze (Turkey)

2010-03-15

163

GaSb Thermophotovoltaic Cells Grown on GaAs Substrate Using the Interfacial Misfit Array Method  

NASA Astrophysics Data System (ADS)

We present gallium antimonide (GaSb) p-i- n photodiodes for use as thermophotovoltaic (TPV) cells grown on gallium arsenide (100) substrates using the interfacial misfit array method. Devices were grown using molecular beam epitaxy and fabricated using standard microfabrication processes. X-ray diffraction was used to measure the strain, and current-voltage ( I- V) tests were performed to determine the photovoltaic properties of the TPV cells. Energy generation at low efficiencies was achieved, and device performance was critically analyzed.

DeMeo, Dante; Shemelya, Corey; Downs, Chandler; Licht, Abigail; Magden, Emir Salih; Rotter, Tom; Dhital, Chetan; Wilson, Stephen; Balakrishnan, Ganesh; Vandervelde, Thomas E.

2014-04-01

164

Behaviour of SH-SY5Y neuroblastoma cell line grown in different media and on different chemically modified substrates.  

PubMed

Among the parameters that can be tested in experiments on neuronal cell culture the use of different culture media and substrates represents a powerful assay to influence cell adhesion and differentiation. In this work, plasma-enhanced-chemical vapour depositions (PE-CVD) from acrylic acid and allylamine vapours have been performed to deposit coatings bearing oxygen (O)- and nitrogen (N)-containing functional groups on polyethylenetherephtalate (PET) surface. Human neuroblastoma SH-SY5Y cells were grown on plasma modified substrates and in presence of media containing different amount of fetal calf serum (FCS) or in serum-free medium containing cAMP. Our results showed that N-containing substrates improved cell adhesion, while the neurites sprouting was influenced by cell culture media. Interestingly, the presence of carboxylic groups on the modified surface can influence the expression of a differentiation marker, neurofilament-200 (NF-H), in cells grown in serum-containing media. PMID:17391751

Buttiglione, M; Vitiello, F; Sardella, E; Petrone, L; Nardulli, M; Favia, P; d'Agostino, R; Gristina, R

2007-07-01

165

Activation of the H+ATPase in the plasma membrane of cells of Saccharomyces cerevisiae grown under mild copper stress  

Microsoft Academic Search

Cells of Saccharomyces cerevisiae exibited a more active plasma membrane H+-ATPase during growth in media supplemented with CuSO4 concentrations equal to or below 1 mM than did cells cultivated in the absence of copper stress. Maximal specific activities\\u000a were found with 0.5 mM CuSO4. ATPase activity declined when cells were grown with higher concentrations up to 1.5 mM (the maximal

Alexandra R. Fernandes; Francisco P. Peixoto; Isabel Sá-Correia

1998-01-01

166

Improved lentiviral gene transfer into human embryonic stem cells grown in co-culture with murine feeder and stroma cells.  

PubMed

Genetic modification of human embryonic stem cells (hESCs) using biophysical DNA transfection methods are hampered by the very low single cell survival rate and cloning efficiency of hESCs. Lentiviral gene transfer strategies are widely used to genetically modify hESCs but limited transduction efficiencies in the presence of feeder or stroma cells present problems, particularly if vesicular stomatitis virus glycoprotein (VSV-G) pseudotyped viral particles are applied. Here, we investigated whether the recently described semen derived enhancer of virus infection (SEVI) and alternative viral envelope proteins derived from either Gibbon ape leukaemia virus (GALV) or feline leukaemia virus (RD114) are applicable for transducing hESCs during co-culture with feeder or stroma cells. Our first set of experiments demonstrates that SEVI has no toxic effect on murine or hESCs and that exposure to SEVI does not interfere with the pluripotency-associated phenotype. Focusing on hESCs, we were able to further demonstrate that SEVI increases the transduction efficiencies of GALV and RD114 pseudotyped lentiviral vectors. More importantly, aiming at targeted differentiation of hESCs into functional somatic cell types, GALV pseudotyped lentiviral particles could efficiently and exclusively transduce hESCs grown in co-culture with OP9-GFP stroma cells (which were often used to induce differentiation into haematopoietic derivatives). PMID:21812756

Wurm, Melanie; Gross, Benjamin; Sgodda, Malte; Ständker, Ludger; Müller, Thomas; Forssmann, Wolf-Georg; Horn, Peter A; Blasczyk, Rainer; Cantz, Tobias

2011-10-01

167

A polarized cell model for Chikungunya virus infection: entry and egress of virus occurs at the apical domain of polarized cells.  

PubMed

Chikungunya virus (CHIKV) has resulted in several outbreaks in the past six decades. The clinical symptoms of Chikungunya infection include fever, skin rash, arthralgia, and an increasing incidence of encephalitis. The re-emergence of CHIKV with more severe pathogenesis highlights its potential threat on our human health. In this study, polarized HBMEC, polarized Vero C1008 and non-polarized Vero cells grown on cell culture inserts were infected with CHIKV apically or basolaterally. Plaque assays, viral binding assays and immunofluorescence assays demonstrated apical entry and release of CHIKV in polarized HBMEC and Vero C1008. Drug treatment studies were performed to elucidate both host cell and viral factors involved in the sorting and release of CHIKV at the apical domain of polarized cells. Disruption of host cell myosin II, microtubule and microfilament networks did not disrupt the polarized release of CHIKV. However, treatment with tunicamycin resulted in a bi-directional release of CHIKV, suggesting that N-glycans of CHIKV envelope glycoproteins could serve as apical sorting signals. PMID:24587455

Lim, Pei Jin; Chu, Justin Jang Hann

2014-02-01

168

A Polarized Cell Model for Chikungunya Virus Infection: Entry and Egress of Virus Occurs at the Apical Domain of Polarized Cells  

PubMed Central

Chikungunya virus (CHIKV) has resulted in several outbreaks in the past six decades. The clinical symptoms of Chikungunya infection include fever, skin rash, arthralgia, and an increasing incidence of encephalitis. The re-emergence of CHIKV with more severe pathogenesis highlights its potential threat on our human health. In this study, polarized HBMEC, polarized Vero C1008 and non-polarized Vero cells grown on cell culture inserts were infected with CHIKV apically or basolaterally. Plaque assays, viral binding assays and immunofluorescence assays demonstrated apical entry and release of CHIKV in polarized HBMEC and Vero C1008. Drug treatment studies were performed to elucidate both host cell and viral factors involved in the sorting and release of CHIKV at the apical domain of polarized cells. Disruption of host cell myosin II, microtubule and microfilament networks did not disrupt the polarized release of CHIKV. However, treatment with tunicamycin resulted in a bi-directional release of CHIKV, suggesting that N-glycans of CHIKV envelope glycoproteins could serve as apical sorting signals.

Lim, Pei Jin; Chu, Justin Jang Hann

2014-01-01

169

Epoxidation of Short-Chain Alkenes by Resting-Cell Suspensions of Propane-Grown Bacteria  

PubMed Central

Sixteen new cultures of propane-utilizing bacteria were isolated from lake water from Warinanco Park, Linden, N.J. and from lake and soil samples from Bayway Refinery, Linden, N.J. In addition, 19 known cultures obtained from culture collections were also found to be able to grow on propane as the sole carbon and energy source. In addition to their ability to oxidize n-alkanes, resting-cell suspensions of both new cultures and known cultures grown on propane oxidize short-chain alkenes to their corresponding 1,2-epoxides. Among the substrate alkenes, propylene was oxidized at the highest rate. In contrast to the case with methylotrophic bacteria, the product epoxides are further metabolized. Propane and other gaseous n-alkanes inhibit the epoxidation of propylene. The optimum conditions for in vivo epoxidation are described. Results from inhibition studies indicate that a propane monooxygenase system catalyzes both the epoxidation and hydroxylation reactions. Experiments with cell-free extracts show that both hydroxylation and epoxidation activities are located in the soluble fraction obtained after 80,000 × g centrifugation.

Hou, Ching T.; Patel, Ramesh; Laskin, Allen I.; Barnabe, Nancy; Barist, Irene

1983-01-01

170

Radial junction amorphous silicon solar cells on PECVD-grown silicon nanowires.  

PubMed

Constructing radial junction hydrogenated amorphous silicon (a-Si:H) solar cells on top of silicon nanowires (SiNWs) represents a promising approach towards high performance and cost-effective thin film photovoltaics. We here develop an all-in situ strategy to grow SiNWs, via a vapour-liquid-solid (VLS) mechanism on top of ZnO-coated glass substrate, in a plasma-enhanced chemical vapour deposition (PECVD) reactor. Controlling the distribution of indium catalyst drops allows us to tailor the as-grown SiNW arrays into suitable size and density, which in turn results in both a sufficient light trapping effect and a suitable arrangement allowing for conformal coverage of SiNWs by subsequent a-Si:H layers. We then demonstrate the fabrication of radial junction solar cells and carry on a parametric study designed to shed light on the absorption and quantum efficiency response, as functions of the intrinsic a-Si:H layer thickness and the density of SiNWs. These results lay a solid foundation for future structural optimization and performance ramp-up of the radial junction thin film a-Si:H photovoltaics. PMID:22539188

Yu, Linwei; O'Donnell, Benedict; Foldyna, Martin; Roca i Cabarrocas, Pere

2012-05-17

171

Radial junction amorphous silicon solar cells on PECVD-grown silicon nanowires  

NASA Astrophysics Data System (ADS)

Constructing radial junction hydrogenated amorphous silicon (a-Si:H) solar cells on top of silicon nanowires (SiNWs) represents a promising approach towards high performance and cost-effective thin film photovoltaics. We here develop an all-in situ strategy to grow SiNWs, via a vapour-liquid-solid (VLS) mechanism on top of ZnO-coated glass substrate, in a plasma-enhanced chemical vapour deposition (PECVD) reactor. Controlling the distribution of indium catalyst drops allows us to tailor the as-grown SiNW arrays into suitable size and density, which in turn results in both a sufficient light trapping effect and a suitable arrangement allowing for conformal coverage of SiNWs by subsequent a-Si:H layers. We then demonstrate the fabrication of radial junction solar cells and carry on a parametric study designed to shed light on the absorption and quantum efficiency response, as functions of the intrinsic a-Si:H layer thickness and the density of SiNWs. These results lay a solid foundation for future structural optimization and performance ramp-up of the radial junction thin film a-Si:H photovoltaics.

Yu, Linwei; O'Donnell, Benedict; Foldyna, Martin; Cabarrocas, Pere Roca i.

2012-05-01

172

Biotransformation of Hydroxylaminobenzene and Aminophenol by Pseudomonas putida 2NP8 Cells Grown in the Presence of 3Nitrophenol  

Microsoft Academic Search

Biotransformation products of hydroxylaminobenzene and aminophenol produced by 3-nitrophenol-grown cells of Pseudomonas putida 2NP8, a strain grown on 2- and 3-nitrophenol, were characterized. Ammonia, 2-aminophenol, 4-aminophenol, 4-benzoquinone, N-acetyl-4-aminophenol, N-acetyl-2-aminophenol, 2-amino- phenoxazine-3-one, 4-hydroquinone, and catechol were produced from hydroxylaminobenzene. Ammonia, N-acetyl-2-aminophenol, and 2-aminophenoxazine-3-one were produced from 2-aminophenol. All of these metabolites were also found in the nitrobenzene transformation medium, and this

JIAN-SHEN ZHAO; AJAY SINGH; XIAO-DONG HUANG; OWEN P. WARD

2000-01-01

173

Pectic Polysaccharide Breakdown of Cell Walls in Cucumber Roots Grown with Calcium Starvation 1  

PubMed Central

Pectic polysaccharides from the roots of cucumber (Cucumis sativus L.) grown in liquid culture medium with or without calcium (1 mm CaCl2) were studied after extraction successively by hot water and Na hexametaphosphate solution. The Ca2+ starvation-treatment caused a striking reduction in content of extracted pectic polysaccharide; from an equivalent weight of cell walls, only 33.1% of the control level was extracted from root cell walls of plants cultured under Ca2+ deficiency. The extracted pectic polysaccharides were fractionated into neutral and acidic polymers by a DEAE-Sephadex column. The acidic polymers, which represented more than 76% of the yield, appeared to be a major fraction of extracted pectic polysaccharides. The changes of molecular size and glycosyl residue composition of this fraction were compared for the control and Ca2+-deprived samples. The results indicate that Ca2+ deficiency caused structural changes which could involve both branching pattern and extent of contiguous galacturonosyl units in the water-solubilized pectic polysaccharides. Ca2+ starvation also led to a notable decrease in molecular size of the hexametaphosphate-solubilized polysaccharides and, to a lesser extent, of the water-solubilized fraction as well. In addition, polygalacturonase activity in tissue homogenates increased remarkably with the Ca2+ deficiency, whereas ?-galactosidase activity did not undergo a change. Thus, it appears that one major effect of Ca2+ deprivation was to stimulate polygalacturonase activity, an effect which could be involved in the control of the breakdown of pectic polysaccharides in the cell walls.

Konno, Haruyoshi; Yamaya, Tomoyuki; Yamasaki, Yoshiki; Matsumoto, Hideaki

1984-01-01

174

Proteomic analysis of Staphylococcus aureus biofilm cells grown under physiologically relevant fluid shear stress conditions  

PubMed Central

Background The biofilm forming bacterium Staphylococcus aureus is responsible for maladies ranging from severe skin infection to major diseases such as bacteremia, endocarditis and osteomyelitis. A flow displacement system was used to grow S. aureus biofilms in four physiologically relevant fluid shear rates (50, 100, 500 and 1000 s-1) to identify proteins that are associated with biofilm. Results Global protein expressions from the membrane and cytosolic fractions of S. aureus biofilm cells grown under the above shear rate conditions are reported. Sixteen proteins in the membrane-enriched fraction and eight proteins in the cytosolic fraction showed significantly altered expression (p?

2014-01-01

175

The Effect of In Vivo Grown Corneal Epithelium Transplantation on Persistent Epithelial Defects with Limbal Stem Cell Deficiency  

PubMed Central

We report our experience with corneal epithelium, grown in vivo, transplantation in three patients with persistent epithelial defect (PED). The three patients had ocular surface disease unresponsive to standard treatments and were therefore chosen for transplantation. They underwent transplantation of epithelial sheets, grown in vivo, to the most affected eye. In vivo cultivation was carried out in the cornea of a living related donor. After epithelialization was completed, the epithelium grown on an amniotic membrane was harvested gently; it was then transplanted into the patient's eye after debridement of fibrovascular tissue. The cultivated epithelium was completely epithelialized by 2 weeks; it was well-differentiated with well-formed hemidesmosome. On immunohistochemical staining, p63, connexin 43, and Integrin ?4 were expressed in the cells on the epithelial sheet. The PED was covered completely and maintained for 4 weeks in all cases. However, corneal erosion recurred after 5 weeks in two cases. This novel technique demonstrates the corneal epithelial cells can be expanded in vivo successfully on denuded amniotic membrane of a healthy cornea and harvested safely. A corneal epithelial sheet, grown in vivo, can be transplanted to treat eye with a severe ocular surface disease, such as total limbal deficiency.

Kim, Jee Taek; Chun, Yeoun Sook; Song, Geo Young

2008-01-01

176

Salicylic acid induces apoptosis in colon carcinoma cells grown in-vitro: Influence of oxygen and salicylic acid concentration  

SciTech Connect

In solid tumors the hypoxic environment can promote tumor progression and resistance to therapy. Recently, acetylsalicylic acid a major component of analgesic drugs and its metabolite salicylic acid (SA) have been shown to reduce the risk of colon cancer, but the mechanisms of action remain still unclear. Here we elucidate the effects of physiologically relevant concentrations of SA on colon carcinoma cells (CaCo-2) grown under normoxic and hypoxic conditions. Western blotting, caspase-3/7 apoptosis assays, MTS cell-proliferation assays, LDH cytotoxicity assays and hydrogen peroxide measurements were performed to investigate the effects of 1 and 10 {mu}M SA on CaCo-2 cells grown under normoxic conditions and cells exposed to hypoxia. Under normoxic conditions, SA did not influence cell proliferation or LDH release of CaCo-2 cells. However, caspase-3/7 activity was significantly increased. Under hypoxia, cell proliferation was reduced and LDH release and caspase-3/7 activities were increased. None of these parameters was altered by the addition of SA under hypoxic conditions. Hypoxia increased hydrogen peroxide concentrations 300-fold and SA significantly augmented the release of hydrogen peroxide under normoxic, but not under hypoxic conditions. Phosphorylation of the pro-survival kinases akt and erk1/2 was not changed by SA under hypoxic conditions, whereas under normoxia SA reduced phosphorylation of erk1/2 after 2 hours. We conclude that in colon carcinoma cells effects of SA on apoptosis and cellular signaling are dependent on the availability of oxygen. -- Highlights: Black-Right-Pointing-Pointer Effects of salicylic acid on colon carcinoma cells grown under normoxic and hypoxic conditions Black-Right-Pointing-Pointer Salicylic acid increases caspase-3/7 activity and hydrogen peroxide release under normoxia Black-Right-Pointing-Pointer Salicylic acid decreases pro-survival erk-1/2 phosphorylation under normoxia Black-Right-Pointing-Pointer Salicylic acid does not influence any of the investigated parameters under hypoxia.

Zitta, Karina; Meybohm, Patrick; Bein, Berthold; Huang, Ying; Heinrich, Christin; Scholz, Jens; Steinfath, Markus; Albrecht, Martin, E-mail: Albrecht@anaesthesie.uni-kiel.de

2012-04-15

177

Characteristics of CuInSe2 Bridgman ingots grown with sodium [solar cell manufacture  

Microsoft Academic Search

Crystals of CuInSe2 were grown by a horizontal Bridgman method for the first time with metallic sodium. The amount of sodium was varied with levels at 0.05, 0.25 and 2 at.% approximately. However, as the sodium content was increased, the crystalline quality decreased. Hot probe measurements showed that the ingots grown with higher sodium content (0.25 and 2 at.%) were

H. P. Wang; I. Shih; C. H. Champness

2000-01-01

178

H+/ATP stoichiometry of cowpea Rhizobium sp. strain 32H1 cells grown under nitrogen-fixing and nitrogen-nonfixing conditions.  

PubMed Central

The obligate aerobe Cowpea Rhizobium sp. strain 32H1 in axenic culture is able to fix N2 when grown under 0.2% O2 but not when grown under 21% O2. It was, therefore, of interest to investigate ATP synthesis in these cells grown under the two conditions. When respiring in buffers having pHs ranging from 6 to 8.5, cells grown under either O2 tension maintained an intracellular pH more alkaline than the exterior. The transmembrane chemical gradient of H+ (delta pH) was essentially the same under both conditions of growth, decreasing from ca. 90 mV at medium pH 6 to ca. 30 mV at pH 8.5. However, the transmembrane electrical gradient (delta psi) was significantly higher in cells grown under 21% O2 (150 to 166 mV) than in cells grown under 0.2% O2, the latter being 16 mV at pH 6 and increasing to 88 mV at pH 8.5. Therefore, the proton motive force of 21% O2-grown cells ranged from 237 mV at external pH 6 to 185 mV at pH 8.5, compared with a proton motive force of 114 to 121 mV in the 0.2% O2-grown cells. The cells grown in 0.2% O2 had the same proton motive force whether tested at 21 or at 0.2% O2. The phosphorylation potential, calculated from the intracellular ATP, ADP, and Pi concentrations, was 424 mV in the 21% O2-grown cells and 436 mV in the 0.2% O2-grown cells. Thus, the 21% O2-grown cells translocated 1.8 to 2.3 H+/ATP synthesized by the H+-ATPase, whereas the H+/ATP ratio for 0.2% O2-grown cells was 3.7 to 3.8.

Gober, J W; Kashket, E R

1984-01-01

179

Guggulsterone production in cell suspension cultures of the guggul tree, Commiphora wightii , grown in shake-flasks and bioreactors  

Microsoft Academic Search

Cell suspension cultures of Commiphora wightii, grown in modified MS medium containing 2,4-dichlorophenoxyacetic acid (0.5 mg l?1) and kinetin (0.25 mg l?1), produced ?5 ?g guggulsterone g?1 dry wt. In a 2 l stirred tank bioreactor, the biomass was 5.5 g l?1 and total guggulsterone was 36 ?g l?1.

Meeta Mathur; K. G. Ramawat

2007-01-01

180

Effect of Se\\/(Ga+In) ratio on MBE grown Cu(In,Ga)Se 2 thin film solar cell  

Microsoft Academic Search

The effect of Se\\/(Ga+In) ratio, expressed as Se\\/III, on the electrical, optical and structural properties of Cu(In,Ga)Se2, abbreviated as CIGS, thin film and on the solar cell performance was studied. CIGS films with various Se\\/III ratios were grown by the molecular beam epitaxy (MBE) process. With decreasing Se\\/III ratio, the resistivity of CIGS film was found to be increased and

M. M. Islam; T. Sakurai; S. Ishizuka; A. Yamada; H. Shibata; K. Sakurai; K. Matsubara; S. Niki; K. Akimoto

2009-01-01

181

Physical properties of InGaAsP/InP grown by molecular beam epitaxy with valve phosphorous cracker cell  

NASA Astrophysics Data System (ADS)

InGaAsP films grown on InP substrate by solid source molecular beam epitaxy (SSMBE) using a valve phosphorous cracker cell are investigated. It is found that the films grown at flux ratios fAs/( fAs+ fp) from 0.45 to 0.50 show a superior quality. It is also found that As pressure plays a crucial role in the scattering process; for the films grown at higher arsenic beam pressure (BEP), the Hall mobility ? is dominated by impurity scattering, polar phonon scattering and alloy scattering. For the films with high quality, optical scattering and alloy scattering dominate the mobility. The exponent of T for the films grown at low BEP is found to be as high as 2.54, which cannot be explained by impurity scattering alone. It is believed that, in addition to the impurity-related scattering, some defects associated with As vacancies also significantly contribute to the scattering, especially at low temperatures.

Zhang, D. H.; Shi, W.; Zheng, H. Q.; Yoon, S. F.; Kam, C. H.; Wang, X. Z.

2000-04-01

182

Accumulation of a novel glycolipid and a betaine lipid in cells of Rhodobacter sphaeroides grown under phosphate limitation.  

PubMed

Cells of the photosynthetic bacterium Rhodobacter sphaeroides grown under phosphate-limiting conditions accumulated nonphosphorous glycolipids and lipids carrying head groups derived from amino acids. Concomitantly, the relative amount of phosphoglycerolipids decreased from 90 to 22 mol% of total polar lipids in the membranes. Two lipids, not detectable in cells grown under standard conditions, were synthesized during phosphate-limited growth. Fast atom bombardment mass spectroscopy, exact mass measurements, 1H NMR spectroscopy, sugar composition analysis, and methylation analysis of the predominant glycolipid led to the identification of the novel compound 1,2-di-O-acyl-3-O-[alpha-D-glucopyranosyl-(1-->4)-O-beta-D-galactopyr anosyl]glycerol. The second lipid was identified as the betaine lipid 1,2-di-O-acyl-[4'-(N,N,N-trimethyl)-homoserine]glycerol by cochromatography employing an authentic standard from Chlamydomonas reinhardtii, fast atom bombardment mass spectroscopy, exact mass measurements, and 1H NMR spectroscopy. Prior to this observation, the occurrence of this lipid was thought to be restricted to lower plants and algae. Apparently, these newly synthesized nonphosphorous lipids, in addition to the sulfo- and the ornithine lipid also found in R. sphaeroides grown under optimal conditions, take over the role of phosphoglycerolipids in phosphate-deprived cells. PMID:7872771

Benning, C; Huang, Z H; Gage, D A

1995-02-20

183

Growth and characterization of Czochralski-grown n and p-type GaAs for space solar cell substrates  

NASA Technical Reports Server (NTRS)

Progress in LEC (liquid encapsulated Czochralski) crystal growth techniques for producing high-quality, 3-inch-diameter, n- and p-type GaAs crystals suitable for solar cell applications is described. The LEC crystals with low dislocation densities and background impurities, high electrical mobilities, good dopant uniformity, and long diffusion lengths were reproducibly grown through control of the material synthesis, growth and doping conditions. The capability for producing these large-area, high-quality substrates should positively impact the manufacturability of highly efficiency, low cost, radiation-hard GaAs solar cells.

Chen, R. T.

1983-01-01

184

75 FR 65581 - Proposed Amendment and Revocation of Class E Airspace, Vero Beach, FL  

Federal Register 2010, 2011, 2012, 2013

...designated as an extension to Class D surface area at Vero Beach Municipal Airport...Class E airspace designated as surface area to remove any reference to the...designated as an extension to Class D surface area to eliminate controlled...

2010-10-26

185

33 CFR 110.73b - Indian River at Vero Beach, Fla.  

Code of Federal Regulations, 2010 CFR

...Areas § 110.73b Indian River at Vero Beach, Fla. (a) Area A. Beginning at a point located on the eastern shore of Fritz Is. at latitude 27°39â²32.5â³ N., longitude 80°22â²20.6â³ W. following the shoreline northward to the northwest...

2010-07-01

186

33 CFR 110.73b - Indian River at Vero Beach, Fla.  

Code of Federal Regulations, 2010 CFR

...Areas § 110.73b Indian River at Vero Beach, Fla. (a) Area A. Beginning at a point located on the eastern shore of Fritz Is. at latitude 27°39â²32.5â³ N., longitude 80°22â²20.6â³ W. following the shoreline northward to the northwest...

2009-07-01

187

Epitaxial Crystal Silicon Absorber Layers and Solar Cells Grown at 1.8 Microns per Minute: Preprint  

SciTech Connect

We have grown device-quality epitaxial silicon thin films at growth rates up to 1.8 ?m/min, using hot-wire chemical vapor deposition from silane at substrate temperatures below 750 degrees C. At these rates, which are more than 30 times faster than those used by the amorphous and nanocrystalline Si industry, capital costs for large-scale solar cell production would be dramatically reduced, even for cell absorber layers up to 10 ?m thick. We achieved high growth rates by optimizing the three key parameters: silane flow, depletion, and filament geometry, based on our model developed earlier. Hydrogen coverage of the filament surface likely limits silane decomposition and growth rate at high system pressures. No considerable deterioration in PV device performance is observed when grown at high rate, provided that the epitaxial growth is initiated at low rate. A simple mesa device structure (wafer/epi Si/a-Si(i)/a-Si:H(p)/ITO) with a 2.3 um epitaxial silicon absorber layer was grown at 700 nm/min. The finished device had an open-circuit voltage of 0.424 V without hydrogenation treatment.

Bobela, D. C.; Teplin, C. W.; Young, D. L.; Branz, H. M.; Stradins, P.

2011-07-01

188

Utilization of metabolic energy under saline conditions: changes in properties of ATP dependent enzymes in plant cells grown under saline conditions  

Microsoft Academic Search

The effect of growth in saline medium on the activity of two ATP utilizing enzymes was studied. Hexokinase in carrot (Daucus\\u000a carota L.) cells grown in suspension culture either in the absence or presence of 150 ml NaCl, and tonoplast H+-ATPase in tobacco (Nicotiana tabacum L. cv. Wisconsin 38) cells grown in suspension culture either in the absence of presence

M. Reuveni

1992-01-01

189

Improved guggulsterone production from sugars, precursors, and morphactin in cell cultures of Commiphora wightii grown in shake flasks and a bioreactor  

Microsoft Academic Search

Cell cultures of Commiphora wightii (Arnott.) Bhandari were grown in shake flasks and a bioreactor and an increase in guggulsterone accumulation up to 18 ?g\\u000a l?1 was recorded in cells grown in the production medium containing a combination of sucrose:glucose (4% total), precursors (phenylalanine,\\u000a pyruvic acid, xylose, and sodium acetate), morphactin, and 2iP. A yield of 10 g l?1 biomass and ?200 ?g

Meeta Mathur; K. G. Ramawat

2008-01-01

190

Biotransformation of d-Limonene to (+) trans-Carveol by Toluene-Grown Rhodococcus opacus PWD4 Cells  

PubMed Central

The toluene-degrading strain Rhodococcus opacus PWD4 was found to hydroxylate d-limonene exclusively in the 6-position, yielding enantiomerically pure (+) trans-carveol and traces of (+) carvone. This biotransformation was studied using cells cultivated in chemostat culture with toluene as a carbon and energy source. The maximal specific activity of (+) trans-carveol formation was 14.7 U (g of cells [dry weight])?1, and the final yield was 94 to 97%. Toluene was found to be a strong competitive inhibitor of the d-limonene conversion. Glucose-grown cells did not form any trans-carveol from d-limonene. These results suggest that one of the enzymes involved in toluene degradation is responsible for this allylic monohydroxylation. Another toluene degrader (Rhodococcus globerulus PWD8) had a lower specific activity but was found to oxidize most of the formed trans-carveol to (+) carvone, allowing for the biocatalytic production of this flavor compound.

Duetz, Wouter A.; Fjallman, Ann H. M.; Ren, Shuyu; Jourdat, Catherine; Witholt, Bernard

2001-01-01

191

Pemetrexed alters folate phenotype and inflammatory profile in EA.hy 926 cells grown under low-folate conditions  

PubMed Central

Elevated homocysteine is a risk marker for several major human pathologies. Emerging evidence suggests that perturbations of folate/homocysteine metabolism can directly modify production of inflammatory mediators. Pemetrexed acts by inhibiting thymidylate synthetase (TYMS), dihydrofolate reductase (DHFR), and glycinamide ribonucleotide formyltransferase (GARFT). EA.hy 926 cells grown under low (“Lo”) and high (“Hi”) folate conditions were treated with pemetrexed. The concentrations of several intracellular folate derivatives were measured using LC-MRM/MS. Lo cells had lower total folate concentrations and a different distribution of the intracellular folate derivatives than Hi cells. Treatment with pemetrexed caused a decrease in individual folate analytes. Microarray analysis showed that several genes were significantly up or down-regulated in pemetrexed treated Lo cells. Several of the significantly up-regulated transcripts were inflammatory. Changes in transcript levels of selected targets, including C3, IL-8, and DHFR, were confirmed by quantitative RT-PCR. C3 and IL-8 transcript levels were increased in pemetrexed-treated Lo cells relative to Lo controls; DHFR transcript levels were decreased. In Lo cells, IL-8 and C3 protein concentrations were increased following pemetrexed treatment. Pemetrexed drug treatment was shown in this study to have effects that lead to an increase in pro-inflammatory mediators in Lo cells. No such changes were observed in Hi cells, suggesting that pemetrexed could not modify the inflammatory profile in the context of cellular folate sufficiency.

Hammons, Andrea L.; Summers, Carolyn M.; Jochems, Jeanine; Arora, Jasbir S.; Zhang, Suhong; Blair, Ian A.; Whitehead, Alexander S.

2014-01-01

192

"allometry" Deterministic Approaches in Cell Size, Cell Number and Crude Fiber Content Related to the Physical Quality of Kangkong (Ipomoea reptans) Grown Under Different Plant Density Pressures  

NASA Astrophysics Data System (ADS)

Kangkong, especially the upland type (Ipomoea reptans) is popularly consumed as a vegetable dish in the South East Asian countries for its quality related to Vitamins (A and C) and crude fiber contents. Higher fiber contents would prevent from the occurrence of colon cancer and diverticular disease. With young stem edible portion, its cell number and size contribute to the stem crude fiber content. The mathematical approach of allometry of cell size, number, and fiber content of stem could be used in determining the 'best' plant density pressure in producing the quality young stem to be consumed. Basically, allometry is the ratio of relative increment (growth or change) rates of two parameters, or the change rate associated to the log of measured variables relationship. Kangkog grown equal or lower than 55 plants m-2 produced bigger individual plant and good quality (physical) kangkong leafy vegetable, but with lower total yield per unit area as compared to those grown at higher densities.

Selamat, A.; Atiman, S. A.; Puteh, A.; Abdullah, N. A. P.; Mohamed, M. T. M.; Zulkeefli, A. A.; Othman, S.

193

Collagen scaffolds with in situ-grown calcium phosphate for osteogenic differentiation of Wharton's jelly and menstrual blood stem cells.  

PubMed

The aim of this research was to investigate the osteogenic differentiation potential of non-invasively obtained human stem cells on collagen nanocomposite scaffolds with in situ-grown calcium phosphate crystals. The foams had 70% porosity and pore sizes varying in the range 50-200 µm. The elastic modulus and compressive strength of the calcium phosphate containing collagen scaffolds were determined to be 234.5 kPa and 127.1 kPa, respectively, prior to in vitro studies. Mesenchymal stem cells (MSCs) obtained from Wharton's jelly and menstrual blood were seeded on the collagen scaffolds and proliferation and osteogenic differentiation capacities of these cells from two different sources were compared. The cells on the composite scaffold showed the highest alkaline phosphatase activity compared to the controls, cells on tissue culture polystyrene and cells on collagen scaffolds without in situ-formed calcium phosphate. MSCs isolated from both Wharton's jelly and menstrual blood showed a significant level of osteogenic activity, but those from Wharton's jelly performed better. In this study it was shown that collagen nanocomposite scaffolds seeded with cells obtained non-invasively from human tissues could represent a potential construct to be used in bone tissue engineering. Copyright © 2012 John Wiley & Sons, Ltd. PMID:22744919

Karadas, Ozge; Yucel, Deniz; Kenar, Halime; Torun Kose, Gamze; Hasirci, Vasif

2014-07-01

194

Characteristics of a Galactose-adapted Sugarcane Cell Line Grown in Suspension Culture 1  

PubMed Central

Although d-galactose is normally toxic to sugarcane (Saccharum sp.) cells, a cell line that grows on 100 mm galactose has been propagated. Nonadapted cells in a medium containing galactose instead of sucrose accumulate UDP-galactose; these cells also have much lower UDP-galactose 4-epimerase (EC 5.1.3.2) activity than do adapted cells. This enzyme may determine whether or not galactose will cause toxicity symptoms to develop. The growth rate of galactose-adapted cells is similar to most cell lines on several other carbohydrates. The galactose-adapted cells are also similar to sucrose stock cells in cell wall composition and sugar phosphate concentrations, but, like the nonadapted cells, accumulate free galactose.

Maretzki, Andrew; Thom, Margaret

1978-01-01

195

Temperature coefficients and radiation induced DLTS spectra of MOCVD grown n(+)p InP solar cells  

NASA Technical Reports Server (NTRS)

The effects of temperature and radiation on n(+)p InP solar cells and mesa diodes grown by metallorganic chemical vapor deposition (MOCVD) were studied. It was shown that MOCVD is capable of consistently producing good quality InP solar cells with Eff greater than 19 percent which display excellent radiation resistance due to minority carrier injection and thermal annealing. It was also shown that universal predictions of InP device performance based on measurements of a small group of test samples can be expected to be quite accurate, and that the degradation of an InP device due to any incident particle spectrum should be predictable from a measurement following a single low energy proton irradiation.

Walters, Robert J.; Statler, Richard L.; Summers, Geoffrey P.

1991-01-01

196

Physiological and morphological studies of rat pheochromocytoma cells (PC12) chemically fused and grown in culture.  

PubMed Central

Cell fusion induced by polyethylene glycol has been used to produce in culture giant multinucleate PC12 cells (up to 300 micron in diameter compared to 10-20 micron for unfused cells). Fused cells, like their unfused counterparts, were found to express various neuronal properties. They contained catecholamines. In the presence of nerve growth factor they extended long processes and expressed Na+, Ca2+, and K+ conductances generally associated with excitable cells. In the absence of nerve growth factor these cells neither grew long processes nor generated Na+-spikes. Other neuronal properties were also observed. Images

O'Lague, P H; Huttner, S L

1980-01-01

197

Heterocellular cultures of pulmonary alveolar epithelial cells grown on laminin-5 supplemented matrix  

Microsoft Academic Search

Summary  The pulmonary alveolar epithelium consists of alveolar type I (AT1) and alveolar type II (AT2) cells. Interactions between\\u000a these two cell types are necessary for alveolar homeostasis and remodeling. These interactions have been difficult to study\\u000a in vitro because current cell culture models of the alveolar epithelium do not provide a heterocellular population of AT1\\u000a and AT2 cells for an

Brant E. Isakson; Gregory J. Seedorf; Richard L. Lubman; Scott Boitano

2002-01-01

198

Infection of a chimpanzee with hepatitis C virus grown in cell culture  

Microsoft Academic Search

Culture supernatant harvested from Daudi cells, a lymphoplastoid cell line, after 58 days of infection with the H77 strain of hepatitis C virus (HCV), was inoculated into a chimpanzee. HCV RNA, as detected by RT-PCR, first appeared in the serum and liver 5 and 6 weeks, respectively, after in- oculation. Peripheral blood mononuclear cells (PBMC) collected on week 7 were

Yohko K. Shimizu; Hiroko Igarashi; Tomoko Kiyohara; Max Shapiro; Doris C. Wong; Robert H. Purcell; Hiroshi Yoshikura

199

25 years of recombinant proteins from reactor-grown cells — Where do we go from here?  

Microsoft Academic Search

The purpose of this review is to describe the current status and to highlight several emerging trends in the manufacture of recombinant therapeutic proteins in cultivated mammalian cells, focusing on Chinese hamster ovary cells as the major production host. Over the past 25 years, specific and volumetric productivities for recombinant cell lines have increased about 20-fold as the result of improvements

David L. Hacker; Maria De Jesus; Florian M. Wurm

2009-01-01

200

Adipose-derived stromal cells grown on a hydroxyapatite scaffold can support hematopoiesis in regenerated bone marrow in vivo.  

PubMed

Osteoblastic cells are a key component of the bone marrow (BM) stem cell niche and help regulate hematopoietic stem cells (HSCs). We have previously demonstrated that adipose-derived stromal cells (ADSCs) can differentiate into both osteogenic and chondrogenic cells in vitro. The current study examined whether the anatomical architecture of the BM could be regenerated in vivo by using ADSCs cultured on a hydroxyapatite (HA) scaffold. ADSCs from GFP transgenic mice were cultured in vitro on an HA scaffold. The scaffold with the attached cells was implanted subcutaneously onto the backs of C57/BL6 (Ly5.2) recipient mice. Lineage-negative (Lin-) Ly5.1 BM cells transduced with a lentiviral vector containing the luciferase (Luc) gene were intravenously administered to the recipient mice after lethal irradiation. Eight weeks after BM transplantation, the scaffolds were removed from the first recipient mice and subcutaneously implanted into lethally irradiated second recipient mice. The biodistribution and kinetics of Luc(+) Ly5.1 cells were monitored by bioluminescence imaging and flow cytometry. Luc(+) hematopoietic cells were present in the scaffolds of the secondary implanted mice for at least 8 months. Subcutaneous injection of G-CSF resulted in wide distribution of bioluminescence signals from the original scaffolds to the whole body. Therefore, BM regenerated using ADSCs grown on an HA scaffold can support HSC populations in vivo, suggesting that a functional BM niche is reconstituted. These results may have a significant impact on the development of therapeutic strategies for various hematopoietic diseases. PMID:24474575

Ueda, Takahiro; Fujita, Atsushi; Ogawa, Rei; Itoh, Yasuhiko; Fukunaga, Yoshitaka; Shimada, Takashi; Migita, Makoto

2014-06-01

201

Dye-sensitized solar cells with vertically aligned TiO2 nanowire arrays grown on carbon fibers.  

PubMed

One-dimensional semiconductor TiO2 nanowires (TNWs) have received widespread attention from solar cell and related optoelectronics scientists. The controllable synthesis of ordered TNW arrays on arbitrary substrates would benefit both fundamental research and practical applications. Herein, vertically aligned TNW arrays in situ grown on carbon fiber (CF) substrates through a facile, controllable, and seed-assisted thermal process is presented. Also, hierarchical TiO2 -nanoparticle/TNW arrays were prepared that favor both the dye loading and depressed charge recombination of the CF/TNW photoanode. An impressive conversion efficiency of 2.48 % (under air mass 1.5 global illumination) and an apparent efficiency of 4.18 % (with a diffuse board) due to the 3D light harvesting of the wire solar cell were achieved. Moreover, efficient and inexpensive wire solar cells made from all-CF electrodes and completely flexible CF-based wire solar cells were demonstrated, taking into account actual application requirements. This work may provide an intriguing avenue for the pursuit of lightweight, cost-effective, and high-performance flexible/wearable solar cells. PMID:24488679

Cai, Xin; Wu, Hongwei; Hou, Shaocong; Peng, Ming; Yu, Xiao; Zou, Dechun

2014-02-01

202

Positioning effects on quantum dot solar cells grown by molecular beam epitaxy  

SciTech Connect

We report current-voltage and spectral response characteristics of high density InAs/GaAs quantum dot (QD) solar cells with different positions where dots are located. The short circuit current density (J{sub sc}), open circuit voltage (V{sub oc}), and external quantum efficiency of these cells under air mass 1.5 are presented and compared with a GaAs reference cell. An extended photoresponse in contrast to the GaAs reference cell was confirmed for all these cells. The effect of inserting QD layers into emitter and base region on device performance is shown. The J{sub sc} is reduced, while the V{sub oc} is maintained. The cell with QDs located toward the base side shows better performance, confirmed by both current-voltage and spectral response measurements.

Zhou, D.; Sharma, G.; Fimland, B. O. [Department of Electronics and Telecommunications, Norwegian University of Science and Technology (NTNU), NO-7491 Trondheim (Norway); Vullum, P. E.; Thomassen, S. F.; Holmestad, R.; Reenaas, T. W. [Department of Physics, Norwegian University of Science and Technology (NTNU), NO-7491 Trondheim (Norway)

2010-02-22

203

Effect of Hypergravity on Localization Calcium Ions in Plant Cells Grown in Vivo and in Vitro  

NASA Astrophysics Data System (ADS)

Using plant callus tissues and Arabidopsis thaliana plants as model systems we have been investigated the effect of hypergravity on the localization and relative content of calcium ions in photosynthesizing cells. The tobacco callus cells in log stage of growth and mesophyll cells from developed A. thaliana leaves were used in the experiments. Plant samples were exposed to hypergravity at 6.5 g, 10g and 14 g for 15-60 min. After centrifugation, dye Fluo-4 was loaded in the control leaves and the centrifuged samples by the standard cytochemical method. Observation of calcium fluorescence was carried out with a laser confocal microscope LSM 5 Pascal at the excitation wave 488 nm (by the argon laser), at emission wavelength 516 nm. The data of the calcium ion distribution and quantification in cells were obtained using software "Pascal" (Carl Zeiss). The effect of hypergravity on redistribution of calcium ions in plant cells has been established. This effect is depended from exposure time and from the value of hypergravity. The cells cultivated in vitro is showed fast response to hypergravity influence. Plasmolysis cells and calcium domains formation have been observed in most of callus cells. This influence was like to that, which was wrote in Funaria hygrometrica protonema cells after 8.5 g influence (Sytnik et al., 1984). Leaf cells of A. thaliana were of less responsively to hypergravity than callus cells. Sytnik K, Kordyum E, Nedukha O. et al. 1984. Plant Cell Under Change of Geophysical Factors. Kiev: Naukova Dumka, 1-134 p.

Nedukha, Olena

204

Method of measuring nitric oxide release by vascular endothelial cells grown in microfluidic channels  

NASA Astrophysics Data System (ADS)

In this paper, a simple and versatile method is presented which enables detection of nitric oxide (NO) released from vascular endothelial cells (ECs) cultured in microfluidic structures. The culturing system and NO measurement method allow cell shape to be controlled in a non-invasive manner using microfluidic structures while NO release is monitored for cell shape versus function studies. The culturing system consists of arrays of polydimethylsiloxane (PDMS) fluidic channels 120 micrometers in depth and ranging from 100 micrometers to 3 mm in width. The number of channels in each array is varied to yield a constant cell culture surface area (75 mm2) independent of channel width. The channel surfaces are collagen-coated and ECs are cultured to confluence within the channels. A cell scraper is then used to scrape extraneous cells cultured between channels, and NO measurements are made 18 to 24 hours later. A chemiluminescence-based sensor system (NOA 280i, Sievers NO Analyzer) is utilized to measure sample NO. Initial results indicate that NO concentrations can be measured from different microfluidic channel-containing samples using this method. It is shown that there is no significant difference in NO concentration derived from channels of different widths even though the degree of cell elongation varies due to physical constraint by microfluidic channel walls. However, cells treated with TNF? release more NO than untreated cells in fluidic channels, which is comparable to the function of ECs cultured in conventional culturing systems such as culturing dishes.

Hosseinpour, S.; Liu, A. C.; Barakat, A. I.; Choy, J. C.; Gray, B. L.

2014-03-01

205

The density of apical cells of dark-grown protonemata of the moss Ceratodon purpureus  

NASA Technical Reports Server (NTRS)

Determinations of plant or algal cell density (cell mass divided by volume) have rarely accounted for the extracellular matrix or shrinkage during isolation. Three techniques were used to indirectly estimate the density of intact apical cells from protonemata of the moss Ceratodon purpureus. First, the volume fraction of each cell component was determined by stereology, and published values for component density were used to extrapolate to the entire cell. Second, protonemal tips were immersed in bovine serum albumin solutions of different densities, and then the equilibrium density was corrected for the mass of the cell wall. Third, apical cell protoplasts were centrifuged in low-osmolarity gradients, and values were corrected for shrinkage during protoplast isolation. Values from centrifugation (1.004 to 1.015 g/cm3) were considerably lower than from other methods (1.046 to 1.085 g/cm3). This work appears to provide the first corrected estimates of the density of any plant cell. It also documents a method for the isolation of protoplasts specifically from apical cells of protonemal filaments.

Schwuchow, J. M.; Kern, V. D.; Wagner, T.; Sack, F. D.

2000-01-01

206

An Easy-To-Handle Microfluidic Device Suitable for Immunohistochemical Procedures in Mammalian Cells Grown Under Flow Conditions  

PubMed Central

Microfluidics, the technology that manipulates small amount of fluids in microscale complex devices, has undergone a remarkable development during the last decade, by targeting a significant range of applications, including biological tests and single-cell analysis, and by displaying many advantages such as reduced reagent consumption, decreased costs and faster analysis. Furthermore, the introduction of microfluidic tools has revolutionized the study of vascular functions, because the controlled three-dimensional environment and the continuous perfusion provided by the microdevice allow simulating the physiological characteristics of the circulatory system. Researchers interested in the study of vascular physiology, however, are often hampered by the difficulty in handling reduced number of cells after growth in these devices. This work shows how to apply different protocols commonly used in biology, such as the immunofluorescence technique, to cells grown in reversibly-bound microfluidic devices, obtaining results comparable to those retrieved under static conditions in multiwells. In this way, we are able to combine the advantages of microfluidic, i.e., application of continuous flow and shear stress, with classical protocols for the study of endothelial cells.

Fede, C.; Fortunati, I.; Petrelli, L.; Guidolin, D.; De Caro, R.; Ferrante, C.; Albertin, G.

2014-01-01

207

Transcriptome profiling in Arabidopsis inflorescence stems grown under hypergravity in terms of cell walls and plant hormones  

NASA Astrophysics Data System (ADS)

Land plants rely on lignified secondary cell walls in supporting their body weight on the Earth. Although gravity influences the formation of the secondary cell walls, the regulatory mechanism of their formation by gravity is not yet understood. We carried out a comprehensive analysis of gene expression in inflorescence stems of Arabidopsis thaliana L. using microarray (22 K) to identify genes whose expression is modulated under hypergravity condition (300 g). Total RNA was isolated from the basal region of inflorescence stems of plants grown for 24 h at 300 g or 1 g. Microarray analysis showed that hypergravity up-regulated the expression of 403 genes to more than 2-fold. Hypergravity up-regulated the genes responsible for the biosynthesis or modification of cell wall components such as lignin, xyloglucan, pectin and structural proteins. In addition, hypergravity altered the expression of genes related to the biosynthesis of plant hormones such as auxin and ethylene and that of genes encoding hormone-responsive proteins. Our transcriptome profiling indicates that hypergravity influences the formation of secondary cell walls by modulating the pattern of gene expression, and that auxin and/or ethylene play an important role in signaling hypergravity stimulus.

Tamaoki, D.; Karahara, I.; Nishiuchi, T.; De Oliveira, S.; Schreiber, L.; Wakasugi, T.; Yamada, K.; Yamaguchi, K.; Kamisaka, S.

2009-07-01

208

Changes in levels of cell wall constituents in wheat seedlings grown under continuous hypergravity conditions  

NASA Astrophysics Data System (ADS)

Effects of continuous hypergravity stimuli on the amounts and composition of cell wall constituents were investigated in wheat shoots. Hypergravity (300 g) treatment for three days after germination increased the net amount of cell wall polysaccharides such as hemicellulose and cellulose, but reduced the shoot elongation. As a result, the amount of cell wall polysaccharides per unit length of shoot increased under hypergravity. The hemicellulose fraction contained polysaccharides in the middle and low molecular mass range (5 kDa-1 MDa) and increased in response to hypergravity. Also, the amounts of arabinose (Ara) and xylose (Xyl), the major sugar components of the hemicellulose fraction, increased under hypergravity conditions. In addition to wall polysaccharides, hypergravity increased the amounts of cell wall-bound phenolic acids, such as ferulic acid (FA) and diferulic acid (DFA). Furthermore, the activity of phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) was enhanced under hypergravity conditions. These results suggest that continuous hypergravity stimulates the synthesis of cell wall constituents, especially hemicellulosic arabinoxylans and cell wall-bound FA and DFA in wheat shoots. The increased PAL activity may promote the formation of FA and DFA. These changes in cell wall architecture may be involved in making rigid and tough cell walls under hypergravity conditions and thereby contribute to the ability of plant to sustain their structures against gravitational stimuli.

Wakabayashi, K.; Soga, K.; Kamisaka, S.; Hoson, T.

209

Compensation mechanism of undoped GaAs films grown by molecular beam epitaxy using an As-valved cracker cell  

NASA Astrophysics Data System (ADS)

We have investigated GaAs films epitaxially grown by molecular beam epitaxy (MBE) using a valved As-cracker source. When the cracking temperature ( Tc) of the As-valved cracker cell is raised, which means the dominant As species changes from As 4 to As 2, the conduction type of the films changed from p - to n -. In order to clarify the origins of the change of compensation mechanism of those GaAs films, estimations were made using electrical (Hall effect) and optical (photoluminescence and far-infrared) measurements. Impurity incorporation behaviors suggested by these estimations are found to give a reasonable explanation for the change of conduction type, that is, the change of compensation mechanism.

Hong, C. U.; Gozu, S.; Okayasu, J.; Koyano, M.; Katayama, S.; Hori, H.; Yamada, S.

1998-06-01

210

Fenofibric Acid Reduces Fibronectin and Collagen Type IV Overexpression in Human Retinal Pigment Epithelial Cells Grown in Conditions Mimicking the Diabetic Milieu: Functional Implications in Retinal Permeability  

PubMed Central

Purpose. To determine whether fenofibric acid (FA) reduces high glucose (HG)–induced basement membrane component overexpression and hyperpermeability in human retinal pigment epithelial (RPE) cells. Methods. Retinal pigment epithelial cells (ARPE-19) were cultured for 18 days in normal glucose (5 mM) or HG (25 mM) medium and studied for the effects of FA on fibronectin (FN) and collagen IV (Coll IV) expression. During last 3 days of the experiment, 100 ?M FA was added to cells grown in HG medium or in HG medium plus IL-1? (HG + IL-1?) to mimic, at least in part, the inflammatory aspect of the diabetic milieu. Real-time RT-PCR was performed to determine FN and Coll IV mRNA levels, whereas protein levels were assessed by Western blot analyses. Cell monolayer morphology and barrier function were analyzed by confocal microscopy using specific antibodies against tight junction proteins, ZO-1, and claudin-1 and by measuring apical-basolateral movements of FITC-dextran, respectively. Results. FN and Coll IV expression were significantly increased in RPE cells grown in HG or HG + IL-1? medium compared with cells grown in normal medium. When cells grown in HG or HG + IL-1? medium were treated with FA, significant reductions in FN and Coll IV expression were observed. In addition, exposure to FA decreased excess permeability in a dose-dependent manner in cells grown in HG + IL-1? medium. This effect was unrelated to changes in tight junction protein content. Conclusions. Findings from this study suggest that the downregulation of basement membrane components by FA may have a protective effect against outer blood-retinal barrier leakage associated with diabetic retinopathy.

Trudeau, Kyle; Roy, Sumon; Guo, Wen; Hernandez, Cristina; Villarroel, Marta; Simo, Rafael

2011-01-01

211

Changes in cell wall architecture of wheat coleoptiles grown under continuous hypergravity conditions  

NASA Astrophysics Data System (ADS)

Modifications of cell wall structure of wheat coleoptiles in response to continuous hypergravity (300 g) treatment were investigated. Length of coleoptiles exposed to hypergravity for 2-4 days from germination stage was 60-70% of that of 1 g control. The net amounts of cell wall polysaccharides, such as hemicellulose and cellulose, of hypergravity-treated coleoptiles increased as much as those of 1 g control coleoptiles during the incubation period. As a result, the levels of cell wall polysaccharides per unit length of coleoptile, which mean the thickness of cell walls, largely increased under hypergravity conditions. Particularly, the amounts of hemicellulosic polymers with middle molecular mass (0.2-1 MDa) largely increased from day 2 to 3 under hypergravity conditions. The major sugar components of the hemicellulose fraction are arabinose, xylose and glucose. The ratios of arabinose and xylose to glucose were higher in hypergravity-treated coleoptiles than in control coleoptiles. The fractionation of hemicellulosic polymers into the neutral and acidic polymers by the anion-exchange column showed that the levels of acidic polymers (mainly composed of arabinoxylans) in cell walls of hypergravity-treated coleoptiles were higher than those of control coleoptiles. In addition to wall polysaccharides, the amounts of cell wall-bound phenolics, such as ferulic acid and diferulic acid, substantially increased during the incubation period both in 1 g control and hypergravity-treated coleoptiles. Especially, the levels of diferulic acid which cross-links hemicellulosic polymers were higher in hypergravity-treated coleoptiles than in control coleoptiles during the incubation period. These results suggest that hypergravity stimuli from the germination stage bias the type of synthesized hemicellulosic polysaccharides, although they do not restrict the net synthesis of cell wall constituents in wheat coleoptiles. The stimulation of the synthesis of arabinoxylans and of the formation of DFA, and also the resultant cell wall thickening may contribute to plant resistance to gravity stimuli.

Wakabayashi, K.; Soga, K.; Kamisaka, S.; Hoson, T.

212

Repair of sublethal damage in two human tumor cell lines grown as multicellular spheroids  

SciTech Connect

Multicellular tumor spheroids (MTS) provide a suitable in vitro model to study radiation sensitivity of tumor cells. Two cell lines of human origin, obtained from a neuroblastoma (NB-100) and a squamous cell carcinoma (HN-1), were exposed to graded doses (4-9 Gy) of radiation with 18 MV photons. Radiation was applied either as a single or as a split dose with an interval of 6 hr to determine the extent of sublethal damage repair. Treated spheroids regrew at approximately the same growth rate as control multicellular tumor spheroids, preceded by a static or regression phase. Radiation response was quantified in terms of regrowth delay, expressed as the time needed for treated spheroids to obtain an 8-fold increase of the initial volume at the time of irradiation. Data obtained from regrowth delay analysis were used to calculate the extent of sublethal damage repair, showing for the squamous cell carcinoma line a fractionally higher capacity to repair sublethal damage than the neuroblastoma line. Repair increased with larger dose fractions in both cell lines. Our results show that multicellular tumor spheroids from the two cell lines used in this study are best applicable at relatively high total radiation doses. This makes multicellular tumor spheroids a suitable model for the in vitro evaluation of clinical treatment rationales such as hyperfractionation.

Schwachoefer, J.H.C.; Crooijmans, R.P.; van Gasteren, J.J.; Hoogenhout, J.; Jerusalem, C.R.; Kal, H.B.; Theeuwes, A.G. (Univ. of Nijmegen (Netherlands))

1989-09-01

213

Repair of sublethal damage in two human tumor cell lines grown as multicellular spheroids.  

PubMed

Multicellular tumor spheroids (MTS) provide a suitable in vitro model to study radiation sensitivity of tumor cells. Two cell lines of human origin, obtained from a neuroblastoma (NB-100) and a squamous cell carcinoma (HN-1), were exposed to graded doses (4-9 Gy) of radiation with 18 MV photons. Radiation was applied either as a single or as a split dose with an interval of 6 hr to determine the extent of sublethal damage repair. Treated spheroids regrew at approximately the same growth rate as control multicellular tumor spheroids, preceded by a static or regression phase. Radiation response was quantified in terms of regrowth delay, expressed as the time needed for treated spheroids to obtain an 8-fold increase of the initial volume at the time of irradiation. Data obtained from regrowth delay analysis were used to calculate the extent of sublethal damage repair, showing for the squamous cell carcinoma line a fractionally higher capacity to repair sublethal damage than the neuroblastoma line. Repair increased with larger dose fractions in both cell lines. Our results show that multicellular tumor spheroids from the two cell lines used in this study are best applicable at relatively high total radiation doses. This makes multicellular tumor spheroids a suitable model for the in vitro evaluation of clinical treatment rationales such as hyperfractionation. PMID:2777647

Schwachöfer, J H; Crooijmans, R P; van Gasteren, J J; Hoogenhout, J; Jerusalem, C R; Kal, H B; Theeuwes, A G

1989-09-01

214

Feasibility of using thin crystalline silicon films epitaxially grown at 165 °C in solar cells: A computer simulation study  

NASA Astrophysics Data System (ADS)

We have previously reported on the successful deposition of heterojunction solar cells whose thin intrinsic crystalline absorber layer is grown using the standard radio frequency plasma enhanced chemical vapour deposition process at 165 °C on highly doped P-type (100) crystalline silicon substrates. The structure had an N-doped hydrogenated amorphous silicon emitter deposited on top of the intrinsic epitaxial silicon layer. However to form the basis of a solar cell, the epitaxial silicon film must be chiefly responsible for the photo-generated current of the structure and not the underlying crystalline silicon substrate. In this article we use detailed electrical-optical modelling to calculate the minimum thickness of the epitaxial silicon layer for this to happen. We have also investigated by modelling the influence of the a-Si:H/epitaxial-Si and epitaxial-Si/c-Si interface defects, the thickness of the epitaxial silicon layer and its volume defect density on cell performance. Finally by varying the input parameters and considering various light-trapping schemes, we show that it is possible to attain a conversion efficiency in excess of 13% using only a 5 micron thick epitaxial silicon layer.

Chakraborty, S.; Cariou, R.; Labrune, M.; Cabarrocas, P. Roca i.; Chatterjee, P.

2013-04-01

215

Comparative analysis of Cryptococcus neoformans acid-resistant particles generated from pigmented cells grown in different laccase substrates.  

PubMed

Cryptococcus neoformans produces pigments in vitro in the presence of exogenous substrate. We characterized acid-resistant particles isolated from pigmented cells grown in L-dopa, methyl-dopa, (-)-epinephrine or (-)-norepinephrine. The goals of this study were to determine whether pigments made from each of these substrates were melanins and the consequences of pigmentation on related cell characteristics. The greatest yield of acid-resistant particles occurred with methyl-dopa followed by L-dopa. Electron microscopy indicated that L-dopa and methyl-dopa produced particles with thicker shells. The mAb 6D2 reacted with all particles, but a lower reactivity was observed with epinephrine-derived particles. ESR analysis revealed that epinephrine-derived particles failed to produce a stable free radical signal typical of melanins. Growth of C. neoformans in different substrates affected cell and capsule size but not capsule induction. Hence, the type of pigment produced by C. neoformans is dependent on the substrate and not all pigments meet the criteria for melanins. PMID:16289955

Garcia-Rivera, Javier; Eisenman, Helene C; Nosanchuk, Joshua D; Aisen, Philip; Zaragoza, Oscar; Moadel, Tiffany; Dadachova, Ekaterina; Casadevall, Arturo

2005-12-01

216

Photovoltaic characteristics of n(+)pp(+) InP solar cells grown by OMVPE  

NASA Technical Reports Server (NTRS)

The photovoltaic characteristics of n(+)/p/p(+) homojunction InP solar cells fabricated by organometallic vapor-phase epitaxy (OMVPE) are described. The cells are characterized by I-V, C-V and quantum efficiency measurements, and simulations are used to obtain various device and material parameters. The I-V characteristics show a high recombination rate in the depletion region; this is shown to be independent of the impurity used. It is shown that cadmium is easier to use as an acceptor for the p base and p(+) buffer and is therefore beneficial. The high quantum efficiency of 98 percent at long wavelengths measured in these cells indicates a very good collection efficiency in the base. The short-wavelength quantum efficiency is poor, indicating a high surface recombination.

Tyagi, S.; Singh, K.; Bhimnathwala, H.; Ghandhi, S. K.; Borrego, J. M.

1990-01-01

217

Space concentrator solar cells based on multilayer LPE grown AlGaAs/GaAs heterostructure  

NASA Technical Reports Server (NTRS)

The high efficiency solar cells based on multilayer AlGaAs/GaAs heterostructures, prepared by low temperature liquid phase epitaxy (LPE), were developed and tested. An investigation of the low temperature LPE process for the crystallization of AlGaAs heterostructures of as high as 24.0 to 24.7 percent under AMO conditions at concentration ratios of 20 to 100x, were reached. Developed solar cells show substantial radiation resistance to the damage induced by 3.75 MeV electrons.

Khvostikov, V. P.; Larionov, V. R.; Paleeva, E. V.; Sorokina, S. V.; Chosta, O. I.; Shvarts, M. Z.; Zimogorova, N. S.

1995-01-01

218

Clonal vaccinia virus grown in cell culture as a new smallpox vaccine  

Microsoft Academic Search

Although the smallpox virus was eradicated over 20 years ago, its potential release through bioterrorism has generated renewed interest in vaccination. To develop a modern smallpox vaccine, we have adapted vaccinia virus that was derived from the existing Dryvax vaccine for growth in a human diploid cell line. We characterized six cloned and one uncloned vaccine candidates. One clone, designated

Jian Liu; Konstantin V Pugachev; Gwendolyn A Myers; Brie Coughlin; Paul S Blum; Richard Nichols; Casey Johnson; John Cruz; Jeffrey S Kennedy; Francis A Ennis; Richard Weltzin; Thomas P Monath

2003-01-01

219

Stable release of BDNF from the fibroblast cell line NIH3T3 grown on silicone elastomers enhances survival of spiral ganglion cells in vitro and in vivo.  

PubMed

The treatment of choice for profound sensorineural hearing loss (SNHL) is direct electrical stimulation of spiral ganglion cells (SGC) via a cochlear implant (CI). The number and excitability of SGC seem to be critical for the success that can be achieved via CI treatment. However, SNHL is associated with degeneration of SGC. Long-term drug delivery to the inner ear for improving SGC survival may be achieved by functionalisation of CI electrodes with cells providing growth factors. Therefore, the capacity of brain-derived neurotrophic factor (BDNF)-secreting NIH3T3 cells grown on cylindrically shaped silicone elastomers (SE) to exert local and sustained neuroprotective effects was assessed in vitro and in vivo. An in vitro model to investigate adhesion and cell growth of lentivirally modified NIH3T3 cells synthesising BDNF on SE was established. The bioactivity of BDNF was characterised by co-cultivation of SGC with cell-coated SE. In addition, cell-coated SE were implanted into deafened guinea pigs. The recombinant NIH3T3 cells proliferated on silicone surfaces during 14 days of cultivation and expressed significantly increasing BDNF levels. Enhanced survival rates and neurite outgrowth of SGC demonstrated the bioactivity of BDNF in vitro. Implantation of SE with adhering BDNF-secreting NIH3T3 cells into the cochleae of systemically deafened guinea pigs induced a significant increase in SGC survival in comparison to SE without cell coating. Our data demonstrate a novel approach of cell-based long-term drug delivery to support SGC survival in vitro and in vivo. This therapeutic strategy--once transferred to cells suitable for clinical application--may improve CI performance. PMID:22564255

Warnecke, Athanasia; Sasse, Susanne; Wenzel, Gentiana I; Hoffmann, Andrea; Gross, Gerhard; Paasche, Gerrit; Scheper, Verena; Reich, Uta; Esser, Karl-Heinz; Lenarz, Thomas; Stöver, Timo; Wissel, Kirsten

2012-07-01

220

Induction of IL8 expression by Cordyceps militaris grown on germinated soybeans through lipid rafts formation and signaling pathways via ERK and JNK in A549 cells  

Microsoft Academic Search

Aim of the studyIn order to elucidate immunoregulatory mechanisms of Cordyceps militaris, a methanol extract of Cordyceps militaris grown on germinated soybeans was prepared and its immunoregulatory effect in the human lung epithelial cells was investigated by examining its ability to induce IL-8 expression.

Ji Young Han; Jintaek Im; Jung Nam Choi; Choong Hwan Lee; Hye Jin Park; Dong Ki Park; Cheol-Heui Yun; Seung Hyun Han

2010-01-01

221

Amorphous silicon solar cells on natively textured ZnO grown by PECVD  

Microsoft Academic Search

Natively textured ZnO layers deposited by the expanding thermal plasma CVD technique between 150 and 350°C at a deposition rate between 0.65 and 0.75 nm\\/s have been investigated with respect to their suitability as front electrode material for amorphous silicon pin solar cells in comparison to reference SnO2:F (Asahi U-type). At higher substrate temperature and with growing thickness, the surface

J. Löffler; R Groenen; J. L Linden; M. C. M van de Sanden; R. E. I Schropp

2001-01-01

222

Hybrid solar cells based on dc magnetron sputtered films of n-ITO on APMOVPE grown p-InP  

NASA Astrophysics Data System (ADS)

Hybrid indium-tin-oxide (ITO)/InP solar cells are discussed. The cells are constructed by dc magnetron sputter deposition of ITO onto high-quality InP films grown by atmospheric pressure metal-organic vapor-phase epitaxy (APMOVPE). A record efficiency of 18.9 percent, measured under standard Solar Energy Research Institute reporting conditions, has been obtained. The p-InP surface is shown to be type converted, principally by the ITO, but with the extent of conversion being modified by the nature of the sputtering gas. The deposition process, in itself, is not responsible for the type conversion. Dark currents have been suppressed by more than three orders of magnitude by the addition of hydrogen to the sputtering gas during deposition of a thin (5 nm) interface layer. Without this layer, and using only the more usual argon/oxygen mixture, the devices had poorer efficiencies and were unstable. A discussion of associated quantum efficiencies and capacitance/voltage measurements is also presented from which it is concluded that further improvements in efficiency will result from better control over the type-conversion process.

Coutts, T. J.; Li, X.; Wanlass, M. W.; Emery, K. A.; Gessert, T. A.

223

Hybrid solar cells based on dc magnetron sputtered films of n-ITO on APMOVPE grown p-InP  

NASA Technical Reports Server (NTRS)

Hybrid indium-tin-oxide (ITO)/InP solar cells are discussed. The cells are constructed by dc magnetron sputter deposition of ITO onto high-quality InP films grown by atmospheric pressure metal-organic vapor-phase epitaxy (APMOVPE). A record efficiency of 18.9 percent, measured under standard Solar Energy Research Institute reporting conditions, has been obtained. The p-InP surface is shown to be type converted, principally by the ITO, but with the extent of conversion being modified by the nature of the sputtering gas. The deposition process, in itself, is not responsible for the type conversion. Dark currents have been suppressed by more than three orders of magnitude by the addition of hydrogen to the sputtering gas during deposition of a thin (5 nm) interface layer. Without this layer, and using only the more usual argon/oxygen mixture, the devices had poorer efficiencies and were unstable. A discussion of associated quantum efficiencies and capacitance/voltage measurements is also presented from which it is concluded that further improvements in efficiency will result from better control over the type-conversion process.

Coutts, T. J.; Li, X.; Wanlass, M. W.; Emery, K. A.; Gessert, T. A.

1988-01-01

224

Effect of Se/(Ga+In) ratio on MBE grown Cu(In,Ga)Se 2 thin film solar cell  

NASA Astrophysics Data System (ADS)

The effect of Se/(Ga+In) ratio, expressed as Se/III, on the electrical, optical and structural properties of Cu(In,Ga)Se 2, abbreviated as CIGS, thin film and on the solar cell performance was studied. CIGS films with various Se/III ratios were grown by the molecular beam epitaxy (MBE) process. With decreasing Se/III ratio, the resistivity of CIGS film was found to be increased and majority carrier hole concentration was decreased. This is probably due to the formation of Se-vacancy and/or vacancy complex. Again, the band-gap energy of CIGS film increased and the Cu content in the bulk of the film decreased with decreasing Se/III ratio despite the Ga/III ratio in the film remained almost unchanged. These results may suggest the formation of anti-site defects such as In Cu in CIGS. Solar cell performance degraded with decreasing Se/III ratio in CIGS absorber layer and the cause of the degradation was discussed.

Islam, M. M.; Sakurai, T.; Ishizuka, S.; Yamada, A.; Shibata, H.; Sakurai, K.; Matsubara, K.; Niki, S.; Akimoto, K.

2009-03-01

225

Moringa oleifera Lam. (Moringaceae) grown in Nigeria: In vitro antisickling activity on deoxygenated erythrocyte cells  

PubMed Central

Context: Traditional medicine, which is more available and affordable for the poor uses medicinal plants for the treatment and management of various ailments, including the sickle cell disease (SCD). About 24 million Nigerians are carriers of this sickled cell gene, while approximately 2.4 million are SCD patients. Moringa oleifera Lam. (Moringaceae) possesses high nutritional value and has been used in folklore medicine to treat various ailments related to pain and inflammation. Chemical, pharmacological and pharmacognostical applications of Moringa oleifera have been reported. Objective: This study investigated the antisickling potential of polar and non-polar extracts of the seed, flower and leaf of Moringa oleifera for the first time. Materials and Methods: Using crude methanol extract, aqueous extract, ethyl acetate and butanol, the in vitro antisickling activities of Moringa oleifera fractions, were evaluated using erythrocyte cells deoxygenated with 2% sodium metabisulphite. p-Hydroxybenzoic acid and normal saline were employed as positive and negative controls. Results: Phytochemical screening revealed the presence of saponins, free anthraquinones, and alkaloids. Extracts of the seed and flower demonstrated a higher (P<0.05) antisickling activity in comparison to the leaf extract. The leaf extract, as well as those of the seed and flower, equally demonstrated a (P<0.05) reversal of sickled erythrocytes. Discussions and Conclusions: These findings suggest that Moringa oleifera may play a role in the management of SCD, by incorporation of its fractions into recipes. More extensive biological evaluations and further studies will be necessary for the chemical characterization of the antisickling principles.

Adejumo, Olufunmilayo E.; Kolapo, Adelodun L.; Folarin, Akintomiwa O.

2012-01-01

226

Genotoxic Effects of Low- and High-LET Radiation on Human Epithelial Cells Grown in 2-D Versus 3-D Culture  

NASA Technical Reports Server (NTRS)

Risk estimation for radiation-induced cancer relies heavily on human epidemiology data obtained from terrestrial irradiation incidents from sources such as medical and occupational exposures as well as from the atomic bomb survivors. No such data exists for exposures to the types and doses of high-LET radiation that will be encountered during space travel; therefore, risk assessment for space radiation requires the use of data derived from cell culture and animal models. The use of experimental models that most accurately replicate the response of human tissues is critical for precision in risk projections. This work compares the genotoxic effects of radiation on normal human epithelial cells grown in standard 2-D monolayer culture compared to 3-D organotypic co-culture conditions. These 3-D organotypic models mimic the morphological features, differentiation markers, and growth characteristics of fully-differentiated normal human tissue and are reproducible using defined components. Cultures were irradiated with 2 Gy low-LET gamma rays or varying doses of high-LET particle radiation and genotoxic damage was measured using a modified cytokinesis block micronucleus assay. Our results revealed a 2-fold increase in residual damage in 2 Gy gamma irradiated cells grown under organotypic culture conditions compared to monolayer culture. Irradiation with high-LET particle radiation gave similar results, while background levels of damage were comparable under both scenarios. These observations may be related to the phenomenon of "multicellular resistance" where cancer cells grown as 3-D spheroids or in vivo exhibit an increased resistance to killing by chemotherapeutic agents compared to the same cells grown in 2-D culture. A variety of factors are likely involved in mediating this process, including increased cell-cell communication, microenvironment influences, and changes in cell cycle kinetics that may promote survival of damaged cells in 3-D culture that would otherwise die or be rendered reproductively inactive in 2-D culture.

Patel, Z. S.; Cucinotta, F. A.; Huff, J. L.

2011-01-01

227

Osteogenesis of large segmental radius defects enhanced by basic fibroblast growth factor activated bone marrow stromal cells grown on non-woven hyaluronic acid-based polymer scaffold  

Microsoft Academic Search

Osteogenesis of large segmental radius defects in a rat model was studied by implanting a biodegradable non-woven hyaluronic acid-based polymer scaffold (Hyaff®11) alone or in combination with bone marrow stromal cells (BMSCs). These cells had been previously grown in vitro in mineralising medium either supplemented with basic fibroblast growth factor (bFGF) or unsupplemented. The healing of bone defects was evaluated

G Lisignoli; M Fini; G Giavaresi; N Nicoli Aldini; S Toneguzzi; A Facchini

2002-01-01

228

Osteoprotegerin (OPG) Production by Cells in the Osteoblast Lineage is Regulated by Pulsed Electromagnetic Fields in Cultures Grown on Calcium Phosphate Substrates  

Microsoft Academic Search

Pulsed electromagnetic fields (PEMF) used clinically to stimulate bone formation enhance the osteogenic effects of BMP-2 on\\u000a human mesenchymal stem cells (MSCs) if the MSCs are grown in osteogenic medium and are cultured on calcium phosphate (CaP)\\u000a surfaces rather than tissue culture polystyrene plastic (TCPS). This study tested if PEMF’s effects on cells in the osteoblast\\u000a lineage are substrate dependent

Zvi Schwartz; Maya Fisher; Christoph H. Lohmann; Bruce J. Simon; Barbara D. Boyan

2009-01-01

229

Nucleus image properties and cell death in MCF10F cells grown on slide substrates differing in nature and size  

Microsoft Academic Search

Summary  The immortalized human breast epithelial cell line MCF-10F is an important tool for studies on experimental tumorigenesis\\u000a induced by drugs, transfected Ha-ras oncogene, and hormones. Considering that many relevant data have thus far been established only for MCF-10F cells cultivated\\u000a on glass, and that there are data showing different cell death ratios for tumorigenic cells obtained from benzo[a]pyrene (BP)-transformed\\u000a MCF-10F

Maria Luiza S. Mello; M. H. Lareef; A. B. Santos; J. Russo; B. C. Vidal

2005-01-01

230

Highly transparent and conductive indium tin oxide thin films for solar cells grown by reactive thermal evaporation at low temperature  

NASA Astrophysics Data System (ADS)

Transparent conductive tin-doped indium oxide (In2O3:Sn, ITO) thin films with various Sn-doping concentrations have been prepared using the low cost reactive thermal evaporation (RTE) technique at a low growth temperature of ~160 °C. The structural characteristics, optical and electrical properties of the ITO thin films were investigated. These polycrystalline ITO films exhibited preferential orientation along (222) plane and possessed low resistivities ranging from 3.51 to 5.71 × 10-4 ? cm. The decreased mobility was attributed to the scattering by ionized and neutral impurities at high doping concentrations. The optimized ITO thin film deposited with 6.0 wt% Sn-doping concentration exhibited a high average transparency of 87 % in the wavelength range of 380-900 nm and a low resistivity of 3.74 × 10-4 ? cm with a high Hall mobility of 47 cm2 V-1s-1. A hydrogenated amorphous silicon and silicon-germanium (a-Si:H/a-SiGe:H) double-junction solar cell fabricated with the RTE-grown ITO electrodes presented a conversion efficiency of 10.51 %.

Du, Jian; Chen, Xin-liang; Liu, Cai-chi; Ni, Jian; Hou, Guo-fu; Zhao, Ying; Zhang, Xiao-dan

2014-04-01

231

Characteristics of InP epitaxial layers grown from solid phosphorus using a valve phosphorus cracker cell  

NASA Astrophysics Data System (ADS)

We report the molecular beam epitaxial (MBE) growth of epitaxial InP using a valve phosphorus cracker cell in a range of cracking zone temperature ( Tcr=875 to 950°C), V/III flux ratio (V/III=1.2 to 9.3) and substrate temperature ( Ts=360 to 500°C). The as-grown epitaxial InP on InP (100) substrate was found to be n-type from Hall measurements. The background electron concentration and mobility exhibited a pronounced dependence on the cracking zone temperature, V/III flux ratio and substrate temperature. Using a cracking zone temperature of 850°C, the highest 77 K electron mobility of 40?900 cm 2/V s was achieved at a V/III ratio of 2.3 at substrate temperature ( Ts) of 440°C. The corresponding background electron concentration was 1.74×10 15 cm -3. The photoluminescence (PL) spectra showed two prominent peaks at 1.384 eV and 1.415 eV, with the intensity of the low energy peak becoming stronger at higher cracking zone temperatures. The PL full-width at half maximum (FWHM) was significantly reduced as the V/III ratio was lowered, indicating an improvement in the optical quality of the samples. The lowest PL FWHM achieved at 5 K was 6.3 meV.

Yoon, S. F.; Zheng, H. Q.

1998-07-01

232

Efficient large-scale generation of functional hepatocytes from mouse embryonic stem cells grown in a rotating bioreactor with exogenous growth factors and hormones  

PubMed Central

Introduction Embryonic stem (ES) cells are considered a potentially advantageous source of hepatocytes for both transplantation and the development of bioartificial livers. However, the efficient large-scale generation of functional hepatocytes from ES cells remains a major challenge, especially for those methods compatible with clinical applications. Methods In this study, we investigated whether a large number of functional hepatocytes can be differentiated from mouse ES (mES) cells using a simulated microgravity bioreactor. mES cells were cultured in a rotating bioreactor in the presence of exogenous growth factors and hormones to form embryoid bodies (EBs), which then differentiated into hepatocytes. Results During the rotating culture, most of the EB-derived cells gradually showed the histologic characteristics of normal hepatocytes. More specifically, the expression of hepatic genes and proteins was detected at a higher level in the differentiated cells from the bioreactor culture than in cells from a static culture. On further growing, the EBs on tissue-culture plates, most of the EB-derived cells were found to display the morphologic features of hepatocytes, as well as albumin synthesis. In addition, the EB-derived cells grown in the rotating bioreactor exhibited higher levels of liver-specific functions, such as glycogen storage, cytochrome P450 activity, low-density lipoprotein, and indocyanine green uptake, than did differentiated cells grown in static culture. When the EB-derived cells from day-14 EBs and the cells’ culture supernatant were injected into nude mice, the transplanted cells were engrafted into the recipient livers. Conclusions Large quantities of high-quality hepatocytes can be generated from mES cells in a rotating bioreactor via EB formation. This system may be useful in the large-scale generation of hepatocytes for both cell transplantation and the development of bioartificial livers.

2013-01-01

233

Variable temperature carrier dynamics in bulk (In)GaAsNSb materials grown by MOVPE for multi-junction solar cells  

NASA Astrophysics Data System (ADS)

III-V multi-junction solar cells are typically based on a triple-junction design that consists of an InGaP top junction, a GaAs middle junction, and a bottom junction that employs a 1 - 1.25 eV material grown on GaAs substrates. The most promising 1 - 1.25 eV material that is currently under extensive investigation is bulk dilute nitride such as (In)GaAsNSb lattice matched to GaAs substrates. The approach utilizing dilute nitrides has a great potential to achieve high performance triple-junction solar cells as recently demonstrated by Wiemer, et al., who achieved a record efficiency of 43.5% from multi-junction solar cells including MBE-grown dilute nitride materials [1]. Although MOVPE is a preferred technique over MBE for III-V multi-junction solar cell manufacturing, MOVPEgrown dilute nitride research is at its infancy compared to MBE-grown dilute nitride. In particular, carrier dynamics studies are indispensible in the optimization of MOVPE materials growth parameters to obtain improved solar cell performance. For the present study, we employed time-resolved photoluminescence (TR-PL) techniques to study carrier dynamics in MOVPE-grown bulk dilute nitride InGaAsN materials (Eg = 1 - 1.25 eV at RT) lattice matched to GaAs substrates. In contrast to our earlier samples that showed high background C doping densities, our current samples grown using different metalorganic precursors at higher growth temperatures showed a significantly reduced background doping density of ~ 1017 /cm3. We studied carrier dynamics in (In)GaAsNSb double heterostructures (DH) with different N compositions at room temperature. Post-growth annealing yielded significant improvements in carrier lifetimes of (In)GaAsNSb double heterostructure (DH) samples. Carrier dynamics at various temperatures between 10 K and RT were also studied from (In)GaAsNSb DH samples including those samples grown on different orientation substrates.

Sin, Yongkun; Lingley, Zachary; LaLumondiere, Stephen; Wells, Nathan; Lotshaw, William; Moss, Steven C.; Kim, Tae Wan; Mawst, Luke J.; Kuech, Thomas F.

2014-03-01

234

Expression Profile of Drug and Nutrient Absorption Related Genes in Madin-Darby Canine Kidney (MDCK) Cells Grown under Differentiation Conditions  

PubMed Central

The expression levels of genes involved in drug and nutrient absorption were evaluated in the Madin-Darby Canine Kidney (MDCK) in vitro drug absorption model. MDCK cells were grown on plastic surfaces (for 3 days) or on Transwell® membranes (for 3, 5, 7, and 9 days). The expression profile of genes including ABC transporters, SLC transporters, and cytochrome P450 (CYP) enzymes was determined using the Affymetrix® Canine GeneChip®. Expression of genes whose probe sets passed a stringent confirmation process was examined. Expression of a few transporter (MDR1, PEPT1 and PEPT2) genes in MDCK cells was confirmed by RT-PCR. The overall gene expression profile was strongly influenced by the type of support the cells were grown on. After 3 days of growth, expression of 28% of the genes was statistically different (1.5-fold cutoff, p < 0.05) between the cells grown on plastic and Transwell® membranes. When cells were differentiated on Transwell® membranes, large changes in gene expression profile were observed during the early stages, which then stabilized after 5–7 days. Only a small number of genes encoding drug absorption related SLC, ABC, and CYP were detected in MDCK cells, and most of them exhibited low hybridization signals. Results from this study provide valuable reference information on endogenous gene expression in MDCK cells that could assist in design of drug-transporter and/or drug-enzyme interaction studies, and help interpret the contributions of various transporters and metabolic enzymes in studies with MDCK cells.

Quan, Yong; Jin, Yisheng; Faria, Teresa N.; Tilford, Charles A.; He, Aiqing; Wall, Doris A.; Smith, Ronald L.; Vig, Balvinder S.

2012-01-01

235

High external quantum efficiency and fill-factor InGaN\\/GaN heterojunction solar cells grown by NH3-based molecular beam epitaxy  

Microsoft Academic Search

High external quantum efficiency (EQE) p-i-n heterojunction solar cells grown by NH3-based molecular beam epitaxy are presented. EQE values including optical losses are greater than 50% with fill-factors over 72% when illuminated with a 1 sun AM0 spectrum. Optical absorption measurements in conjunction with EQE measurements indicate an internal quantum efficiency greater than 90% for the InGaN absorbing layer. By

J. R. Lang; C. J. Neufeld; C. A. Hurni; S. C. Cruz; E. Matioli; U. K. Mishra; J. S. Speck

2011-01-01

236

Properties of an nC:P\\/pSi carbon-based photovoltaic cell grown by radio frequency plasma-enhanced chemical vapor deposition at room temperature  

Microsoft Academic Search

The phosphorus-doped amorphous carbon (n-C:P) films were grown by radiofrequency (RF) power-assisted plasma-enhanced chemical vapor deposition (PECVD) at room temperature using a solid phosphorus target. The influence of phosphorus doping on the material properties of n-C:P based on the results of simultaneous characterization are reported. Moreover, solar cell properties such as series resistance, short-circuit current density, open-circuit current voltage, fill

M. Rusop; T. Soga; T. Jimbo

2006-01-01

237

Characteristics of flexible indium tin oxide electrode grown by continuous roll-to-roll sputtering process for flexible organic solar cells  

Microsoft Academic Search

The preparation and characteristics of flexible indium tin oxide (ITO) electrodes grown on polyethylene terephthalate (PET) substrates using a specially designed roll-to-roll sputtering system for use in flexible organic solar cells are described. It was found that both electrical and optical properties of the flexible ITO electrode were critically dependent on the Ar\\/O2 flow ratio in the continuous roll-to-roll sputter

Kwang-Hyuk Choi; Jin-A Jeong; Jae-Wook Kang; Do-Guen Kim; Jong Kuk Kim; Seok-In Na; Dong-Yu Kim; Seok-Soon Kim; Han-Ki Kim

2009-01-01

238

Changes of ribulose bisphosphate carboxylase/oxygenase content, ribulose bisphosphate concentration, and photosynthetic activity during adaptation of high-CO/sub 2/ grown cells to low-CO/sub 2/ conditions in Chlorella pyrenoidosa  

SciTech Connect

Changes of some photosynthetic properties of high-CO/sub 2/ grown cells of Chlorella pyrenoidosa during adaptation to low-CO/sub 2/ conditions have been investigated. The K/sub m/ value of photosynthesis of the high-CO/sub 2/ grown cells for dissolved inorganic carbon was 3.3 millimolar and decreased to 25 to 30 micromolar within 4 hours after transferring to air. In the presence of saturating CO/sub 2/ concentrations the photosynthetic activity of the high-CO/sub 2/ grown cells was 1.5 times as high as that of the low-CO/sub 2/ grown cells. There was a significant rise of the photosynthetic activity during adaptation of the high-CO/sub 2/ grown cells to air, followed by a steady decrease. The activity of ribulose 1,5-bisphosphate carboxylase/oxygenase in both the high and low-CO/sub 2/ grown cells was close to the photosynthetic activity of the cells. The concentration of ribulose 1,5-bisphosphate (RuBP) was higher in the low-CO/sub 2/ adapting and low-CO/sub 2/ grown celsl than in the high-CO/sub 2/ grown cells regardless of the photosynthetic rate. This seems to be due to an increased RuBP regeneration activity during adaptation followed by maintenance of the new higher concentration. The RuBP level always exceeded the concentration of ribulose 1,5-bisphosphate carboxylase/oxygenase RuBP binding sites in both the high- and low-CO/sub 2/ grown cells at any dissolved inorganic carbon concentration.

Yokota, A.; Canvin, D.T.

1986-02-01

239

Bioremoval and recovery of Cd(II) by Pseudoalteromonas sp. SCSE709-6: Comparative study on growing and grown cells.  

PubMed

Comparison of the bioremoval and recovery of Cd(II) by growing and grown marine bacterium Pseudoalteromonas sp. SCSE709-6 was performed in batch systems. Bioremoval with growing cells (Sorption I) showed better performance at low Cd(II) concentrations, whereas bioremoval with grown cells (Sorption II) had significant advantages in both removal efficiency and time consumption at high Cd(II) concentrations. The optimal pH was higher for Sorption I than for Sorption II for achieving the maximum Cd(II) removal efficiency. Complete desorption was achieved using either Na2EDTA or HNO3 as eluent. Cd(II) adsorbed on grown cells had higher tendency to be desorbed. Na2EDTA was a preferable eluent for the recycling biomaterials, whereas HNO3 performed better for the final security disposal of sludge. For Sorption II, both Langmuir and Freundlich isotherms well explained the biosorption behavior, and the pseudo-second-order model better expressed biosorption and desorption kinetics. PMID:24565875

Zhou, Weizhi; Liu, Dongsheng; Zhang, Hai'ou; Kong, Wenqian; Zhang, Yuzhong

2014-08-01

240

Expression of prostasome-like granules by the prostate cancer cell lines PC3, Du145 and LnCaP grown in monolayer.  

PubMed

Prostasomes are a granular type of secretory product in the human prostate gland cells. It is not known, whether in vitro grown cells derived from human prostate cancers also express prostate secretory components containing granules with properties similar to the prostasomes. Therefore, we carried out the present investigation and found that cytospins of in vitro grown PC3, DU145 and LNCaP cells generally expressed a granular secretion. DU145 demonstrated the highest ratio of cells with granules (about 90%), while cytospins of PC3 and LNCaP contained less stained cells (50-70%). Purified granules from PC3 cells were immunoreactive with a monoclonal antibody (mAb78) originally raised against human seminal prostasomes. The PC3 granules also shared the property with human seminal prostasomes having an elevated UV260/UV280 absorbance ratio. On the other hand we found a low aminopeptidase activity in PC3 granules contrary to that of human prostasomes. Prostasomes may form a heterogeneous group with different properties due to the source from which they are isolated and perhaps it is justified to recognize them as different members of a prostasome family. PMID:10680953

Nilsson, B O; Lennartsson, L; Carlsson, L; Nilsson, S; Ronquist, G

1999-01-01

241

Charge photogeneration in hybrid solar cells: a comparison between quantum dots and in situ grown CdS.  

PubMed

We demonstrate that blend films containing poly(3-hexylthiophene-2,5-diyl) and in situ grown CdS display a greater yield of photogenerated charges than a blend containing an equivalent amount of pre-synthesised CdS quantum dots. Moreover, we show that the greater charge yield in the in situ grown films leads to an improvement in device efficiency. The present findings also appear to suggest that charge photogeneration at the CdS/polymer heterojunction is facilitated by the formation of nanoparticle networks as a result of CdS aggregation. PMID:22307222

Reynolds, Luke X; Lutz, Thierry; Dowland, Simon; MacLachlan, Andrew; King, Simon; Haque, Saif A

2012-03-01

242

Magnesium doping of efficient GaAs and Ga(0.75)In(0.25)As solar cells grown by metalorganic chemical vapor deposition  

NASA Technical Reports Server (NTRS)

Magnesium has been substituted for zinc in GaAs and Ga(0.75)In(0.25)As solar cells grown by metalorganic chemical vapor deposition (MOCVD). Bis(cyclopentadienyl)magnesium (Cp2Mg) is used as the MOCVD transport agent for Mg. Full retention of excellent material quality and efficient cell performance results. The substitution of Mg for Zn would enhance the abruptness and reproducibility of doping profiles, and facilitate high temperature processing and operation, due to the much lower diffusion coefficient of Mg, relative to Zn, in these materials.

Lewis, C. R.; Ford, C. W.; Werthen, J. G.

1984-01-01

243

Transport and photoluminescence of silicon-doped GaInP grown by a valved phosphorus cracker cell in solid source molecular beam epitaxy  

NASA Astrophysics Data System (ADS)

We report the transport and photoluminescence (PL) properties of silicon-doped GaInP layers grown on GaAs (100) substrate using a valved phosphorus cracker cell in solid source molecular beam epitaxy. Within the range of silicon (Si) effusion cell temperature investigated (900-1200 °C), the highest electron concentration obtained was 7.7×1018 and 3.2×1018 cm-3 at room temperature and 77 K, respectively. The concentration decreased with further increase in the silicon cell temperature. The Hall mobility at 300 K varied from 356 to 1720 cm2/V s within the range of electron concentration measured (4.5×1016-7.7×1018 cm-3). Except for the sample grown at the highest silicon cell temperature (1200 °C), the PL spectrum of other samples showed a dominant peak attributed to Si donor-to-band transition (D-B), which shifted to higher energy following an increase in the electron concentration. This phenomenon was attributed to the Burstein-Moss effect. The blueshift of the (D-B) transition peak at increasing temperature was attributed to thermal ionization of the Si donors. The sample grown at the highest Si cell temperature showed a PL peak at ~1.913 eV which was attributed to transition between the conduction band and Si acceptor (B-A), with an activation energy of ~57.2 meV as deduced from the PL spectrum. Temperature-dependent Hall measurements confirmed the amphoteric behavior of the Si dopant in this sample. The PL intensity at 10 K decreased and the full width at half maximum increased significantly from ~8 to ~32 meV following an increase in the electron concentration from 4.5×1016 to 7.7×1018 cm-3.

Yoon, S. F.; Mah, K. W.; Zheng, H. Q.

1999-05-01

244

The gene expression profiles of canine mammary cancer cells grown with carcinoma-associated fibroblasts (CAFs) as a co-culture in vitro  

PubMed Central

Background It is supposed that fibroblasts present in tumour microenvironment increase cancer invasiveness and its ability to metastasize but the mechanisms have not been clearly defined yet. Thus, the current study was designed to assess changes in gene expression in five various cancer cell lines grown as a co-culture with the carcinoma-associated fibroblasts (CAFs) in vitro. Results A carcinoma-associated fibroblast cell line was isolated from a canine mammary cancer. Then, a co-culture of cancer cells with the CAFs was established and maintained for 72 hrs. Having sorted the cells, a global gene expression in cancer cells using DNA microarrays was examined. The analysis revealed an up-regulation of 100 genes and a down-regulation of 106 genes in the cancer cells grown as a co-culture with the CAFs in comparison to control conditions. The PANTHER binomial statistics tool was applied to determine statistically over-manifested pathways (p < 0.05). Bulk of the up-regulated genes are involved in the adhesion, the angiogenesis, the epithelial-mesenchymal transition (EMT) and generally take part in the developmental processes. These results were further confirmed using real-time qPCR. Moreover, a wound-healing assay and growth characteristics on Matrigel matrix showed that CAFs increase cancer cell migration and matrix invasion. Conclusion The results of the current study showed that the co-culturing of cancer cells and the CAFs caused significant changes to the cancer gene expression. The presence of the CAFs in a microenvironment of cancer cells promotes adhesion, angiogenesis and EMT.

2012-01-01

245

PROCUSTE1 Encodes a Cellulose Synthase Required for Normal Cell Elongation Specifically in Roots and Dark-Grown Hypocotyls of Arabidopsis  

PubMed Central

Mutants at the PROCUSTE1 (PRC1) locus show decreased cell elongation, specifically in roots and dark-grown hypocotyls. Cell elongation defects are correlated with a cellulose deficiency and the presence of gapped walls. Map-based cloning of PRC1 reveals that it encodes a member (CesA6) of the cellulose synthase catalytic subunit family, of which at least nine other members exist in Arabidopsis. Mutations in another family member, RSW1 (CesA1), cause similar cell wall defects in all cell types, including those in hypocotyls and roots, suggesting that cellulose synthesis in these organs requires the coordinated expression of at least two distinct cellulose synthase isoforms.

Fagard, Mathilde; Desnos, Thierry; Desprez, Thierry; Goubet, Florence; Refregier, Guislaine; Mouille, Gregory; McCann, Maureen; Rayon, Catherine; Vernhettes, Samantha; Hofte, Herman

2000-01-01

246

Comparison of GaInNAs and GaInNAsSb solar cells grown by plasma-assisted molecular beam epitaxy  

NASA Astrophysics Data System (ADS)

We compare dilute nitride GaInNAs and GaInNAsSb solar cells grown by molecular beam epitaxy. Single junction p-i-n diode solar cells were fabricated to test the dilute nitride and antimonide material fabrication process. Triple-junction solar cells were fabricated to test the behavior of single GaInNAs(Sb) junctions in multi-junction configuration. When nitrogen was added to the growth of GaInNAs, good crystal quality was maintained up to 4% of nitrogen at the used growth conditions. Short circuit current densities of the devices could be increased by adding Sb to the growth but at the same time the open circuit voltages decreased due to bandgap shrinkage induced by Sb. In multijunction configuration, the samples with Sb showed inferior properties to ones without it. Lower currents and voltages of Sb-containing cells may be linked to segregation of Sb and transfer to the upper junctions.

Aho, Arto; Tukiainen, Antti; Korpijärvi, Ville-Markus; Polojärvi, Ville; Salmi, Joel; Guina, Mircea

2012-10-01

247

Power recovery of radiation damaged MOCVD grown indium phosphide on silicon solar cells through argon-ion laser annealing. Master`s thesis  

SciTech Connect

This thesis reports the results of a laser annealing technique used to remove defect sites from radiation damaged indium phosphide on silicon MOCVD grown solar cells. This involves the illumination of damaged solar cells with a continuous wave laser to produce a large forward-biased current. The InP/Si cells were irradiated with 1 MeV electrons to a given fluence, and tested for degradation. Light from an argon laser was used to illuminate four cells with an irradiance of 2.5 W/sq cm, producing a current density 3 to 5 times larger than AMO conditions. Cells were annealed at 19 deg C with the laser and at 25 deg C under AMO conditions. Annealing under laser illumination of n/p-type cells resulted in recovery of 48%. P/n type cells lost 4 to 12% of the assumed degradaton. Annealing under AMO conditions resulted in power recovery of 70% in n/p type cells. P/n-type cells recovered approximately 16% of lost power. Results indicate that significant power recovery results from the annealing of defects within n/p type InP/Si solar cells.

Boyer, L.L.

1996-06-01

248

P-cracker cell temperature effects on the optical properties of AlGaInP:Be layers grown by SSMBE  

NASA Astrophysics Data System (ADS)

The optical properties of Be-doped Al 0.2Ga 0.3In 0.5P layers grown on GaAs by solid source molecular beam epitaxy have been studied. In particular, we investigated a set of heavy doped samples grown at different phosphorous cracking temperatures (PCT). The analysis of the 15 K-PL spectra showed three strong transitions below the AlGaInP band edge associated to Be acceptor levels (A 0, X), shallow impurities (A, X) and a broad signal related with oxygen deep levels (O, DL). The Photoluminescence (PL) spectra from samples dramatically change as the PCT is increased, in such a way that in the spectrum from the sample grown at the highest P-cracking zone temperature, the (O, DL) intensity is visibly dominant. Besides, we found that despite that the Be cell temperature being maintained at 1015 °C for all the samples, the Be-doping concentration is reduced as the PCT is increased. Therefore, as PCT increases, the active Be-concentration decreases as a consequence of Be-compensation with O. This phenomena is reflected in the PL properties of the samples as a reduction of the (A 0, X) intensity. The rocking curves of the (0 0 4) planes obtained by high resolution X-ray diffraction (HRXRD) justify the inclusion of impurities and the reduction of the crystal quality of the sample grown at the highest PCT. This work shows that the incorporation of defects during the growth of the AlGaInP:Be films due to O contamination can be reduced using low PCT.

Soubervielle-Montalvo, C.; Hernández, I. C.; Sheldon, M.; Gorbatchev, A. Yu.; Rodríguez, A. G.; de Anda, F.; Zamora-Peredo, L.; Méndez-García, V. H.

2007-04-01

249

Human umbilical cord Wharton's jelly stem cells undergo enhanced chondrogenic differentiation when grown on nanofibrous scaffolds and in a sequential two-stage culture medium environment.  

PubMed

The current treatments used for osteoarthritis from cartilage damage have their disadvantages of donor site morbidity, complicated surgical interventions and risks of infection and graft rejection. Recent advances in tissue engineering have offered much promise in cartilage repair but the best cell source and in vitro system have not as yet been optimised. Human bone marrow mesenchymal stem cells (hBMSCs) have thus far been the cell of choice. However, we derived a unique stem cell from the human umbilical cord Wharton's jelly (hWJSC) that has properties superior to hBMSCs in terms of ready availability, prolonged stemness characteristics in vitro, high proliferation rates, wide multipotency, non-tumorigenicity and tolerance in allogeneic transplantation. We observed enhanced cell attachment, cell proliferation and chondrogenesis of hWJSCs over hBMSCs when grown on PCL/Collagen nanoscaffolds in the presence of a two-stage sequential complex/chondrogenic medium for 21 days. Improvement of these three parameters were confirmed via inverted optics, field emission scanning electron microscopy (FESEM), MTT assay, pellet diameters, Alcian blue histology and staining, glycosaminglycans (GAG) and hyaluronic acid production and expression of key chondrogenic genes (SOX9, Collagen type II, COMP, FMOD) using immunohistochemistry and real-time polymerase chain reaction (qRT-PCR). In separate experiments we demonstrated that the 16 ng/ml of basic fibroblast growth factor (bFGF) present in the complex medium may have contributed to driving chondrogenesis. We conclude that hWJSCs are an attractive stem cell source for inducing chondrogenesis in vitro when grown on nanoscaffolds and exposed sequentially first to complex medium and then followed by chondrogenic medium. PMID:21671058

Fong, Chui-Yee; Subramanian, Arjunan; Gauthaman, Kalamegam; Venugopal, Jayarama; Biswas, Arijit; Ramakrishna, Seeram; Bongso, Ariff

2012-03-01

250

Differences in excitation energy transfer of Arthrospira platensis cells grown in seawater medium and freshwater medium, probed by time-resolved fluorescence spectroscopy  

NASA Astrophysics Data System (ADS)

Excitation energy transfer of Arthrospira platensis cells grown in f/2 medium (a high salinity medium) and SOT medium (a control) was investigated by steady-state and time-resolved spectroscopies. Growth in f/2 medium induced changes in absorption and fluorescence spectra as well as in the energy transfer pathways. Excitation energy captured by phycobilisome (PBS) was transferred directly to photosystem (PS) I, instead of being first transferred to an intermediate (PBS ? PSII ? PSI), as observed in SOT medium. The respiration rate increased while photosynthetic rate reduced in f/2 medium. Possible causes of the differences in light-harvesting and energy-transfer processes between the two media are discussed.

Arba, Muhammad; Aikawa, Shimpei; Niki, Kenta; Yokono, Makio; Kondo, Akihiko; Akimoto, Seiji

2013-11-01

251

Grown-in defects and defects produced by 1-Me electron irradiated in Al0.3Ga0.7As P-N junction solar cells  

NASA Technical Reports Server (NTRS)

Studies of grown-in defects and defects produced by the one-MeV electron irradiation in Al sub 0.3 Ga sub 0.7As p-n junction solar cells fabricated by liquid phase epitaxial (LPE) technique were made for the unirradiated and one-MeV electron irradiated samples, using DLTS and C-V methods. Defect and recombination parameters such as energy level, defect density, carrier capture cross sections and lifetimes were determined for various growth, annealing, and irradiation conditions.

Li, S. S.; Teng, K. W.; Schoenfeld, D. W.; Rahilly, W. P.

1982-01-01

252

Activity of plasma membrane H+ATPase and expression of PMA1 and PMA2 genes in Saccharomyces cerevisiae cells grown at optimal and low pH  

Microsoft Academic Search

Cells of Saccharomyces cerevisiae grown in media with an initial pH of 2.5–6.0, acidified with a strong acid (HCl), exhibited the highest plasma membrane H+-ATPase-specific activity at an initial pH of 6.0. At a lower pH (above pH 2.5) ATPase activity (62–83% of the maximum level)\\u000a still allowed optimal growth. At pH 2.5, ATPase activity was about 30% of the

V. Carmelo; P. Bogaerts; I. Sá-Correia

1996-01-01

253

Synthesis and application of TiO2 single-crystal nanorod arrays grown by multicycle hydrothermal for dye-sensitized solar cells  

NASA Astrophysics Data System (ADS)

TiO2 is a wide band gap semiconductor with important applications in photovoltaic cells. Vertically aligned TiO2 nanorod arrays (NRs) are grown on the fluorine-doped tin oxide (FTO) substrates by a multicycle hydrothermal synthesis process. The samples are characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), high-resolution transmission electron microscopy (HRTEM), and selected-area electron diffraction (SAED). It is found that dye-sensitized solar cells (DSSCs) assembled by the as-prepared TiO2 single-crystal NRs exhibit different trends under the condition of different nucleation and growth concentrations. Optimum cell performance is obtained with high nucleation concentration and low growth cycle concentration. The efficiency enhancement is mainly attributed to the improved specific surface area of the nanorod.

Zhu, Jian-Jing; Zhao, Yu-Long; Zhu, Lei; Gu, Xiu-Quan; Qiang, Ying-Huai

2014-04-01

254

Mitochondria Increase Three-Fold and Mitochondrial Proteins and Lipid Change Dramatically in Postmeristematic Cells in Young Wheat Leaves Grown in Elevated CO2.  

PubMed Central

A dramatic stimulation in mitochondrial biogenesis during the very early stages of leaf development was observed in young wheat plants (Triticum aestivum cv Hereward) grown in elevated CO2 (650 [mu]L L-1). An almost 3-fold increase in the number of mitochondria was observed in the very young leaf cells at the base of the first leaf of a 7-d-old wheat plant. In the same cells large increases in the accumulation of a mitochondrial chaperonin protein and the mitochondrial 2-oxoglutarate dehydrogenase complex and pyruvate dehydrogenase complex were detected by immunolabeling. Furthermore, the basal segment also shows a large increase in the rate of radiolabeling of diphosphatidylglycerol, a lipid confined to the inner mitochondrial membrane. This dramatic response in very young leaf cells to elevated CO2 suggests that the numerous documented positive effects of elevated CO2 on wheat leaf development are initiated as early as 12 h postmitosis.

Robertson, E. J.; Williams, M.; Harwood, J. L.; Lindsay, J. G.; Leaver, C. J.; Leech, R. M.

1995-01-01

255

Microtubules are stabilized in confluent epithelial cells but not in fibroblasts  

PubMed Central

Rhodamine-tagged tubulin was microinjected into epithelial cells (MDCK) and fibroblasts (Vero) to characterize the dynamic properties of labeled microtubules in sparse and confluent cells. Fringe pattern fluorescence photobleaching revealed two components with distinct dynamic properties. About one-third of the injected tubulin diffused rapidly in the cytoplasm with a diffusion coefficient of 1.3-1.6 x 10(- 8) cm2/s. This pool of soluble cytoplasmic tubulin was increased to greater than 80% when cells were treated with nocodazole, or reduced to approximately 20% upon treatment of cells with taxol. Fluorescence recovery of the remaining two-thirds of labeled tubulin occurred with an average half-time (t1/2) of 9-11 min. This pool corresponds to labeled tubulin associated with microtubules, since it was sensitive to treatment of cells with nocodazole and since taxol increased its average t1/2 to greater than 22 min. Movement of photobleached microtubules in the cytoplasm with rates of several micrometers per minute was shown using very small interfringe distances. A significant change in the dynamic properties of microtubules occurred when MDCK cells reached confluency. On a cell average, microtubule half-life was increased about twofold to approximately 16 min. In fact, two populations of cells were detected with respect to their microtubule turnover rates, one with a t1/2 of approximately 9 min and one with a t1/2 of greater than 25 min. Correspondingly, the rate of incorporation of microinjected tubulin into interphase microtubules was reduced about twofold in confluent MDCK cells. In contrast to the MDCK cells, no difference in microtubule dynamics was observed in sparse and confluent populations of Vero fibroblasts, where the average microtubule half- life was approximately 10 min. Thus, microtubules are significantly stabilized in epithelial but not fibroblastic cells grown to confluency.

1990-01-01

256

A vero cell derived combined vaccine against sheep pox and Peste des Petits ruminants for sheep.  

PubMed

The combined sheep pox and Peste des Petits ruminants (PPR) vaccine was prepared in lyophilized form containing recommended doses of both vaccine viruses. Safety and immunogenicity of this combined vaccine was evaluated in sheep. Sheep immunized subcutaneously with 1ml of live attenuated vaccine consisting of 10(3)TCID(50) each of sheep pox virus (SPV) Romanian Fanar (RF) strain and Peste des Petits ruminants virus (PPRV-Sungri/96 strain) were monitored for clinical and serological responses for a period of four weeks post immunization (pi) and two week post challenge (pc). Specific antibodies directed to sheep pox virus could be demonstrated by indirect ELISA and serum neutralization test (SNT). Competitive ELISA and SNT were used for demonstration of antibodies to PPR virus. All the immunized animals resisted challenge with virulent SPV or PPRV on day 30pi, while control animals developed characteristic signs of disease. Specific virus could be detected in the unvaccinated control animals after challenge but not from any of the immunized sheep. Combined vaccine was found to be safe and potent as evident from sero conversion as well as challenge studies in sheep. This indicates that component vaccines did not interfere each other and can be used in target population for economic vaccination strategies. PMID:19428860

Chaudhary, S S; Pandey, K D; Singh, R P; Verma, P C; Gupta, P K

2009-04-28

257

Reduction of the ochratoxin A-induced cytotoxicity in Vero cells by aspartame  

Microsoft Academic Search

Ochratoxin A (OTA) is a mycotoxin produced by Aspergillus ochraceus as well as other moulds. This mycotoxin contaminates animal feed and human food and is nephrotoxic for all animal species\\u000a studied so far. OTA is immunosuppressive, genotoxic, teratogenic and carcinogenic. Recently lipid peroxidation induced by\\u000a OTA has been reported. OTA, a structural analogue of phenylalanine, inhibits protein synthesis by competition

I. Baudrimont; A. M. Betbeder; E. E. Creppy

1997-01-01

258

Antiherpevirus activity of Artemisia arborescens essential oil and inhibition of lateral diffusion in Vero cells  

Microsoft Academic Search

BACKGROUND: New prophylactic and therapeutic tools are needed for the treatment of herpes simplex virus infections. Several essential oils have shown to possess antiviral activity in vitro against a wide spectrum of viruses. AIM: The present study was assess to investigate the activities of the essential oil obtained from leaves of Artemisia arborescens against HSV-1 and HSV-2 METHODS: The cytotoxicity

Manuela Saddi; Adriana Sanna; Filippo Cottiglia; Lorenza Chisu; Laura Casu; Leonardo Bonsignore; Alessandro De Logu

2007-01-01

259

HSV infected RAJI-cells specify HSV specific immediate early and\\/or early DNA-binding proteins  

Microsoft Academic Search

Summary Herpes Simplex Virus specified DNA-binding proteins were purified from virus infected VERO and RAJI cells. The expression of the proteins differed depending on the type of the host cell. Polypeptides corresponding to HSV-1 specific ICP 8 and HSV-2 specific ICSP 11\\/12 were the major products of HSV-infected VERO cells. In RAJI cells polypeptides with a corresponding molecular weight, together

M. Lehtinen

1986-01-01

260

Characterization of silicon-doped InP grown by solid-source molecular beam epitaxy using a valved phosphorus cracker cell  

NASA Astrophysics Data System (ADS)

Silicon (Si)-doped InP layers were grown by solid-source molecular beam epitaxy and characterized by Hall and low-temperature photoluminescence (PL) measurements. During the growth the Si effusion cell temperature was varied from 900 to 1200°C, and the electron concentration measured was between 3.5×10 16 cm -3 and 1.1×10 20 cm -3 at room temperature. The corresponding Hall mobility varied from 3670 to 442 cm 2/V s and 10 400 to 475 cm 2/V s at 300 and 77 K, respectively for different Si cell temperatures. The mobility values were found to be higher compared to n-type InP layers grown by other techniques. 5 K PL spectra showed two peaks for the undoped and low-doped InP layers corresponding to the neutral donor-bound exciton transitions (D 0-X) and the acceptor-related transitions (D-A), respectively. When the Si doping level was increased, the high-energy near-band recombination underwent a typical evolution exhibiting a broadened asymmetric peak, and the peak shifted to high energy that was attributed to the band filling effect. This peak shift compared well with the theoretical prediction based on the Burstein-Moss shift and the band gap narrowing effect.

Radhakrishnan, K.; Zheng, H. Q.; Zhang, P. H.; Yoon, S. F.; Ng, G. I.

1999-07-01

261

Establishment of a macrophagelike cell line derived from U-937, human histiocytic lymphoma, grown serum-free  

Microsoft Academic Search

Summary  A human macrophagelike cell line which grows in serum-free medium was established from a histiocytic lymphoma cell line, U-937.\\u000a U-937 cells failed to differentiate into macrophagelike cells in serum-free medium plus 12-O-tetradecanoyl phorbol 13-acetate (TPA). Fibronectin and albumin in serum were necessary for differentiation of U-937 ceds\\u000a into macrophagelike cells in enriched RDF medium supplemented with insulin, transferrin, ethanolamine, selenite,

Zwe-Ling Kong; Misao Miwa; Hiroki Murakami; Kazuki Shinohara

1990-01-01

262

The increase of choline acetyltransferase activity by docosahexaenoic acid in NG108-15 cells grown in serum-free medium is independent of its effect on cell growth.  

PubMed

We investigated the influence of the polyunsaturated docosahexaenoic acid (22:6n-3; DHA) on the constitutive expression of choline acetyltransferase (ChAT) in native and induced expression in differentiated cholinergic cells NG108-15 grown in serum-free medium. Elimination of serum-derived trophic support resulted in growth arrest and a strong decrease of ChAT activity. In either conditions, DHA largely rescued general indicators of cell growth and function, and partially prevented the decrease of ChAT activity. However, the maximal effect on general cell state in native and differentiated cells, and ChAT activity in native cells, was reached at or below 10 mumol/l of DHA. In contrast, maximal induction of ChAT activity in differentiated cells required about six times higher concentrations of DHA. These data thus demonstrate stimulatory effect of DHA on ChAT activity that is independent of its general cell protective properties. PMID:17004129

Machová, Eva; Málková, Barbora; Lisá, Vera; Nováková, Jana; Dolezal, Vladimír

2006-10-01

263

Identification of chikungunya virus interacting proteins in mammalian cells.  

PubMed

Identification and characterization of virus host interactions is an essential step for the development of novel antiviral strategies. Very few studies have been targeted towards identification of chikungunya virus (CHIKV) interacting host proteins. In current study, virus overlay protein binding assay (VOPBA) and matrix-assisted laser desorption/ ionization time of flight analysis (MALDI TOF/TOF) were employed for the identification of CHIKV binding proteins in mammalian cells. HSP70 and actin were identified as virus binding proteins in HEK-293T and Vero-E6 cells, whereas STAT-2 was identified as an additional protein in Vero-E6 cells. Pre-incubation with anti-HSP70 antibody and miRNA silencing of HSP70 significantly reduced the CHIKV production in HEK-293T and Vero-E6 cells at early time points. These results suggest that CHIKV exploits the housekeeping molecules such as actin, HSP70 and STAT-2 to establish infection in the mammalian cells. PMID:24845503

Paingankar, Mandar S; Arankalle, Vidya A

2014-06-01

264

Establishment of a macrophagelike cell line derived from U-937, human histiocytic lymphoma, grown serum-free.  

PubMed

A human macrophagelike cell line which grows in serum-free medium was established from a histiocytic lymphoma cell line, U-937. U-937 cells failed to differentiate into macrophagelike cells in serum-free medium plus 12-O-tetradecanoyl phorbol 13-acetate (TPA). Fibronectin and albumin in serum were necessary for differentiation of U-937 cells into macrophagelike cells in enriched RDF medium supplemented with insulin, transferrin, ethanolamine, selenite, egg yolk lipoprotein (eRDF-ITESL medium). The established cell line exhibited several characteristic properties of macrophage such as nitroblue tetrazolium reduction, phagocytic and alpha-naphthylbutyrate-esterase activities, and tumor necrosis factor and interleukin 1 production. At present the cells have been continuously maintained in eRDF-ITESL medium through over 150 passages. PMID:2243057

Kong, Z L; Miwa, M; Murakami, H; Shinohara, K

1990-10-01

265

Development of crystalline peroxisomes in methanol-grown cells of the yeast Hansenula polymorpha and its relation to environmental conditions  

Microsoft Academic Search

The development of peroxisomes has been studied in cells of the yeast Hansenula polymorpha during growth on methanol in batch and chemostat cultures. During bud formation, new peroxisomes were generated by the separation of small peroxisomes from mature organelles in the mother cells. The number of peroxisomes migrating to the buds was dependent upon environmental conditions. Aging of cells was

M. Veenhuis; J. P. van Dijken; S. A. F. Pilon; W. Harder

1978-01-01

266

Comparison of human nasal epithelial cells grown as explant outgrowth cultures or dissociated tissue cultures in vitro.  

PubMed

The purpose of this study was to compare cell growth characteristics, ciliated cell differentiation, and function of human nasal epithelial cells established as explant outgrowth cultures or dissociated tissue cultures. Human nasal mucosa of the uncinate process was obtained by endoscopy and epithelial cell cultures were established by explant outgrowth or dissociated tissue culture methods. Epithelial cell growth characteristics were observed by inverted phase contrast microscopy. Ciliated cell differentiation was detected by ?-tubulin IVand ZO-1 immunocytochemistry. Basal and ATP-stimulated ciliary beat frequency (CBF) was measured using a highspeed digital microscopic imaging system. Both the explant and dissociated tissue cultures established as monolayers with tight junctions and differentiated cell composition, with both types of cultures comprising ciliated and non-ciliated epithelial cells. Fibroblasts were also frequently found in explant cultures but rarely seen in dissociated tissue cultures. In both culture systems, the highest ciliated cell density appeared at 7th-10th culture day and declined with time, with the lifespan of ciliated cells ranging from 14 to 21 days. Overall, 10% of the cells in explant cultures and 20% of the cells in the dissociated tissue cultures were ciliated. These two cultures demonstrated similar ciliary beat frequency values at baseline (7.78 ± 1.99 Hz and 7.91 ± 2.52 Hz, respectively) and reacted equivalently following stimulation with 100 ?M ATP. The results of this study indicate that both the explant outgrowth and dissociated tissue culture techniques are suitable for growing well-differentiated nasal ciliated and non-ciliated cells, which have growth characteristics and ciliary activity similar to those of nasal epithelial cells in vivo. PMID:24062261

Jiao, Jian; Meng, Na; Wang, Hong; Zhang, Luo

2013-12-01

267

Effects of phosphorous beam equivalent pressure on GaInAsP/GaAs grown by solid source molecular beam epitaxy with a valve phosphorous cracker cell  

NASA Astrophysics Data System (ADS)

We report the growth and characterization of GaInAsP films on GaAs substrates by solid source molecular beam epitaxy (SSMBE) using a valve phosphorous cracker cell at varied white phosphorous beam equivalent pressure (BEP). It is found that the GaInAsP/GaAs can be easily grown with the solid sources, and the incorporated phosphorous composition as a function of the beam equivalent pressure ratio, R= fP/( fP+ fAs), can be well described by a parabolic relationship. With the increase of the incorporated phosphorous composition, the GaP-, InP-, InAs- and GaAs-like phonon modes shift towards opposite directions and their emission intensities also change. The first three modes shift to larger wave numbers while the last one shifts to smaller wave number. The lattice mismatch, ? a/ a, of the materials grown with varied phosphorous BEP follows a linear relationship. Photoluminescence (PL) measurements reveal that as the phosphorous BEP ratio increases, the peak position or energy band gap of the material shifts towards higher energy; the full-width at half-maximum (FWHM) becomes narrower, and the luminescence intensity becomes higher. In addition, the materials also show smooth surfaces that do not change significantly with phosphorous beam equivalent pressure.

Wang, X. Z.; Zhang, D. H.; Zheng, H. Q.; Yoon, S. F.; Kam, C. H.; Shi, W.; Raman, A.

2000-03-01

268

Structural complexity of non-acid glycosphingolipids in human embryonic stem cells grown under feeder-free conditions.  

PubMed

Due to their pluripotency and growth capability, there are great expectations for human embryonic stem cells, both as a resource for functional studies of early human development and as a renewable source of cells for use in regenerative medicine and transplantation. However, to bring human embryonic stem cells into clinical applications, their cell surface antigen expression and its chemical structural complexity have to be defined. In the present study, total non-acid glycosphingolipid fractions were isolated from two human embryonic stem cell lines (SA121 and SA181) originating from leftover in vitro fertilized human embryos, using large amounts of starting material (1 × 10(9) cells/cell line). The total non-acid glycosphingolipid fractions were characterized by antibody and lectin binding, mass spectrometry, and proton NMR. In addition to the globo-series and type 1 core chain glycosphingolipids previously described in human embryonic stem cells, a number of type 2 core chain glycosphingolipids (neo-lactotetraosylceramide, the H type 2 pentaosylceramide, the Le(x) pentaosylceramide, and the Le(y) hexaosylceramide) were identified as well as the blood group A type 1 hexaosylceramide. Finally, the mono-, di-, and triglycosylceramides were characterized as galactosylceramide, glucosylceramide, lactosylceramide, galabiaosylceramide, globotriaosylceramide, and lactotriaosylceramide. Thus, the glycan diversity of human embryonic stem cells, including cell surface immune determinants, is more complex than previously appreciated. PMID:23404501

Barone, Angela; Benktander, John; Ångström, Jonas; Aspegren, Anders; Björquist, Petter; Teneberg, Susann; Breimer, Michael E

2013-04-01

269

Global gene expression profiles of canine macrophages and canine mammary cancer cells grown as a co-culture in vitro  

PubMed Central

Background Solid tumours comprise various cells, including cancer cells, resident stromal cells, migratory haemopoietic cells and other. These cells regulate tumour growth and metastasis. Macrophages constitute probably the most important element of all interactions within the tumour microenvironment. However, the molecular mechanism, that guides tumour environment, still remains unknown. Exploring the underlying molecular mechanisms that orchestrate these phenomena has been the aim of our study. A co-culture of canine mammary cancer cells and macrophages was established and maintained for 72 hrs. Having sorted the cells, gene expression in cancer cells and macrophages, using DNA microarrays, was examined. The results were confirmed using real-time qPCR and confocal microscopy. Moreover, their ability for migration and invasion has been assessed. Results Microarray analysis showed that the up-regulated genes in the cancer cell lines are involved in 15 highly over-manifested pathways. The pathways that drew our diligent attention included: the inflammation pathway mediated by chemokine and cytokine, the Toll receptor signalling pathway and the B cell activation. The up-regulated genes in the macrophages were involved in only 18 significantly over-manifested pathways: the angiogenesis, the p53 pathway feedback loops2 and the Wnt signalling pathway. The microarray analysis revealed that co-culturing of cancer cells with macrophages initiated the myeloid-specific antigen expression in cancer cells, as well as cytokine/chemokine genes expression. This finding was confirmed at mRNA and protein level. Moreover, we showed that macrophages increase cancer migration and invasion. Conclusions The presence of macrophages in the cancer environment induces acquisition of the macrophage phenotype (specific antigens and chemokines/cytokines expression) in cancer cells. We presumed that cancer cells also acquire other myeloid features, such as: capabilities of cell rolling, spreading, migration and matrix invasion (what has also been confirmed by our results). It may, perhaps, be the result of myeloid-cancer cell hybrid formation, or cancer cells mimicking macrophages phenotype, owing to various proteins secreted by macrophages.

2012-01-01

270

Effects of Taurolidine and Chlorhexidine on SaOS-2 Cells and Human Gingival Fibroblasts Grown on Implant Surfaces.  

PubMed

Purpose: The purpose of the study was the evaluation of possible cytologic effects of taurolidine to fibroblasts and osteoblast-like cells. Materials and Methods: Human gingival fibroblasts and SaOS-2 cells were seeded on samples with sand-blasted and acid-etched surfaces. Both groups were treated with taurolidine, chlorhexidine, and pure water with three different treatment times. Three dates of measurements were set to evaluate cell viability, cytotoxicity, and apoptosis. Results: Highest cytotoxicity was measured in both cell lines in the groups treated with chlorhexidine, while cell viability was lower than in the corresponding taurolidine and pure water groups; on days 3 and 6 these differences were significant. Taurolidine showed similar results to the pure water groups. Conclusion: The results of this study indicate that taurolidine is biocompatible and gentle to the tested human cells for the application time of a mouthrinse. PMID:24818214

John, Gordon; Becker, Jürgen; Schwarz, Frank

2014-01-01

271

Effects of radiation of InP cells epitaxially grown on Si and GaAs substrates  

Microsoft Academic Search

The properties of heteroepitaxial InP solar cells were determined both before and after 10 MeV proton irradiations. Numerical values, obtained for the diffusion and recombination components of the reverse saturation currents, were found to be consistent with the distribution of dislocations. The radiation resistance of the heteroepitaxial cells was significantly greater than that observed for n\\/p homoepitaxial InP cells. The

I. Weinberg; C. K. Swartz; D. J. Brinker; D. M. Wilt

1990-01-01

272

Effects of radiation of InP cells epitaxially grown on Si and GaAs substrates  

Microsoft Academic Search

The properties of heteroepitaxial InP cells were determined both before and after 10-MeV proton irradiations. Numerical values, obtained for the diffusion and recombination components of the reverse saturation currents, were found to be consistent with the distribution of dislocations. The radiation resistance of the heteroepitaxial cells was significantly greater than that observed for n\\/p homoepitaxial InP cells. The carrier removal

I. Weinberg; C. K. Swartz; D. J. Brinker; D. M. Wilt

1990-01-01

273

Effect of calcium restriction on cardenolide accumulation in two cell lines of Digitalis thapsi grown under different light regimes  

Microsoft Academic Search

The effect of calcium-deprivation on growth and the production of cardenolides in two undifferentiated cell lines of Digitalis thapsi maintained under three different light regimes (16 h photoperiod, darkness, or continuous light) was investigated. Growth\\u000a was stimulated by continuous light in both cell lines cultured in complete medium. The light regime did not affect cardenolide\\u000a accumulation in the cells of

Margarita Cacho; Margarita Morán; Purificación Corchete; Jorge Fernández-Tárrago

1999-01-01

274

Human RPE Stem Cells Grown into Polarized RPE Monolayers on a Polyester Matrix Are Maintained after Grafting into Rabbit Subretinal Space  

PubMed Central

Summary Transplantation of the retinal pigment epithelium (RPE) is being developed as a cell-replacement therapy for age-related macular degeneration. Human embryonic stem cell (hESC) and induced pluripotent stem cell (iPSC)-derived RPE are currently translating toward clinic. We introduce the adult human RPE stem cell (hRPESC) as an alternative RPE source. Polarized monolayers of adult hRPESC-derived RPE grown on polyester (PET) membranes had near-native characteristics. Trephined pieces of RPE monolayers on PET were transplanted subretinally in the rabbit, a large-eyed animal model. After 4 days, retinal edema was observed above the implant, detected by spectral domain optical coherence tomography (SD-OCT) and fundoscopy. At 1 week, retinal atrophy overlying the fetal or adult transplant was observed, remaining stable thereafter. Histology obtained 4 weeks after implantation confirmed a continuous polarized human RPE monolayer on PET. Taken together, the xeno-RPE survived with retained characteristics in the subretinal space. These experiments support that adult hRPESC-derived RPE are a potential source for transplantation therapies.

Stanzel, Boris V.; Liu, Zengping; Somboonthanakij, Sudawadee; Wongsawad, Warapat; Brinken, Ralf; Eter, Nicole; Corneo, Barbara; Holz, Frank G.; Temple, Sally; Stern, Jeffrey H.; Blenkinsop, Timothy A.

2014-01-01

275

Photocurrent enhancement in In0.53Ga0.47As solar cells grown on InP/SiO2/Si transferred epitaxial templates  

NASA Astrophysics Data System (ADS)

InP/Si engineered substrates formed by wafer bonding and layer transfer have the potential to significantly reduce the cost and weight of III-V compound semiconductor solar cells. InP/Si substrates were prepared by He implantation of InP prior to bonding to a thermally oxidized Si substrate and annealing to exfoliate an InP thin film. Following thinning of the transferred InP film to remove surface damage caused by the implantation and exfoliation process, InGaAs solar cells lattice-matched to bulk InP were grown on these substrates using metal-organic chemical vapor deposition. The photovoltaic current-voltage characteristics of the InGaAs cells fabricated on the wafer-bonded InP/Si substrates were comparable to those synthesized on commercially available epi-ready InP substrates, and had a ~20% higher short-circuit current which we attribute to the high reflectivity of the InP/SiO2/Si bonding interface. This work provides an initial demonstration of wafer-bonded InP/Si substrates as an alternative to bulk InP substrates for solar cell applications.

Zahler, James M.; Tanabe, Katsuaki; Ladous, Corinne; Pinnington, Tom; Newman, Frederick D.; Atwater, Harry A.

2007-09-01

276

Human RPE Stem Cells Grown into Polarized RPE Monolayers on a Polyester Matrix Are Maintained after Grafting into Rabbit Subretinal Space.  

PubMed

Transplantation of the retinal pigment epithelium (RPE) is being developed as a cell-replacement therapy for age-related macular degeneration. Human embryonic stem cell (hESC) and induced pluripotent stem cell (iPSC)-derived RPE are currently translating toward clinic. We introduce the adult human RPE stem cell (hRPESC) as an alternative RPE source. Polarized monolayers of adult hRPESC-derived RPE grown on polyester (PET) membranes had near-native characteristics. Trephined pieces of RPE monolayers on PET were transplanted subretinally in the rabbit, a large-eyed animal model. After 4 days, retinal edema was observed above the implant, detected by spectral domain optical coherence tomography (SD-OCT) and fundoscopy. At 1 week, retinal atrophy overlying the fetal or adult transplant was observed, remaining stable thereafter. Histology obtained 4 weeks after implantation confirmed a continuous polarized human RPE monolayer on PET. Taken together, the xeno-RPE survived with retained characteristics in the subretinal space. These experiments support that adult hRPESC-derived RPE are a potential source for transplantation therapies. PMID:24511471

Stanzel, Boris V; Liu, Zengping; Somboonthanakij, Sudawadee; Wongsawad, Warapat; Brinken, Ralf; Eter, Nicole; Corneo, Barbara; Holz, Frank G; Temple, Sally; Stern, Jeffrey H; Blenkinsop, Timothy A

2014-01-14

277

Density and length of stomatal and epidermal cells in "living fossil" trees grown under elevated CO 2 and a polar light regime  

NASA Astrophysics Data System (ADS)

During the Cretaceous and early Tertiary, when the climate was warm and the atmospheric CO 2 concentration ([CO 2]) was at least double that of the present-day, polar forests populated high latitude landmasses. We investigated the density and length of stomata and other epidermal cells of two deciduous and three evergreen "living fossil" tree species representative of these ancient forests. These tree species were grown in a simulated Cretaceous high latitude environment at either ambient (400 ppmv) or elevated (800 ppmv) [CO 2] during four years. After 4 years growing at elevated [CO 2], the leaf stomatal density and index (percentage of leaf epidermal cells that are stomata) of these plants were similar to those of their counterparts growing at ambient [CO 2]. While the CO 2 enrichment only modified the stomatal pore length in two of the five studied species, it increased significantly the overall length of the epidermal cells of all the species, reducing their density. These results revealed that leaf epidermal cells of these "living fossil" species were more sensitive than stomata to an experimental doubling of atmospheric CO 2 concentration.

Ogaya, R.; Llorens, L.; Peñuelas, J.

2011-07-01

278

Chemical and Biological Properties of B16 Murine Melanoma Cells Grown in Defined Medium Containing Bovine Serum Albumin1  

Microsoft Academic Search

SUMMARY The addition of 1% (w\\/v) bovine serum albumin (BSA) to the F12 medium utilized for the growth of the B16 mela noma cells significantly stimulated the growth of this cell line. The synthesis of mucopolysaccharides and sialogly- copeptides in this medium is identical with that in Eagle's minimal essential medium with Earle's balanced salt solu tion supplemented with 2

John R. Banks; V. P. Bhavanandan; E. A. Davidson

1977-01-01

279

High external quantum efficiency and fill-factor InGaN/GaN heterojunction solar cells grown by NH{sub 3}-based molecular beam epitaxy  

SciTech Connect

High external quantum efficiency (EQE) p-i-n heterojunction solar cells grown by NH{sub 3}-based molecular beam epitaxy are presented. EQE values including optical losses are greater than 50% with fill-factors over 72% when illuminated with a 1 sun AM0 spectrum. Optical absorption measurements in conjunction with EQE measurements indicate an internal quantum efficiency greater than 90% for the InGaN absorbing layer. By adjusting the thickness of the top p-type GaN window contact layer, it is shown that the short-wavelength (<365 nm) quantum efficiency is limited by the minority carrier diffusion length in highly Mg-doped p-GaN.

Lang, J. R.; Hurni, C. A.; Cruz, S. C.; Matioli, E.; Speck, J. S. [Department of Materials, University of California, Santa Barbara, California 93106 (United States); Neufeld, C. J.; Mishra, U. K. [Department of Electrical and Computer Engineering, University of California, Santa Barbara, California 93106 (United States)

2011-03-28

280

Lipid content and fatty acid composition of green algae Scenedesmus obliquus grown in a constant cell density apparatus  

NASA Technical Reports Server (NTRS)

The lipids of alga Scenedesmus obliquus grown under controlled conditions were separated and fractionated by column and thin-layer chromatography, and fatty acid composition of each lipid component was studied by gas-liquid chromatography (GLC). Total lipids were 11.17%, and neutral lipid, glycolipid and phospholipid fractions were 7.24%, 2.45% and 1.48% on a dry weight basis, respectively. The major neutral lipids were diglycerides, triglycerides, free sterols, hydrocarbons and sterol esters. The glycolipids were: monogalactosyl diglyceride, digalactosyl diglyceride, esterified sterol glycoside, and sterol glycoside. The phospholipids included: phosphatidyl choline, phosphatidyl glycerol and phosphatidyl ethanolamine. Fourteen fatty acids were identified in the four lipid fractions by GLC. The main fatty acids were C18:2, C16:0, C18:3(alpha), C18:1, C16:3, C16:1, and C16:4. Total unsaturated fatty acid and essential fatty acid compositions of the total algal lipids were 80% and 38%, respectively.

Choi, K. J.; Nakhost, Z.; Barzana, E.; Karel, M.

1987-01-01

281

Cell wall changes in nisin-resistant variants of Listeria innocua grown in the presence of high nisin concentrations  

Microsoft Academic Search

Two nisin-resistant variants of a strain of Listeria innocua were isolated after growth in the presence of 500 and 4000 IU ml?1 of nisin A showed increased cell wall hydrophobicity, resistance to phage attack and three different cell wall-acting antibiotics, as well as to the peptidoglycan hydrolytic enzymes lysozyme and mutanolysin, as compared to the parental strain. Transmission electron microscopy

Sophie Maisnier-Patin; Jean Richard

1996-01-01

282

Behaviour of SH-SY5Y neuroblastoma cell line grown in different media and on different chemically modified substrates  

Microsoft Academic Search

Among the parameters that can be tested in experiments on neuronal cell culture the use of different culture media and substrates represents a powerful assay to influence cell adhesion and differentiation. In this work, plasma-enhanced-chemical vapour depositions (PE-CVD) from acrylic acid and allylamine vapours have been performed to deposit coatings bearing oxygen (O)- and nitrogen (N)-containing functional groups on polyethylenetherephtalate

M. Buttiglione; F. Vitiello; E. Sardella; L. Petrone; M. Nardulli; P. Favia; R. d’Agostino; R. Gristina

2007-01-01

283

Effects of radiation of InP cells epitaxially grown on Si and GaAs substrates  

NASA Technical Reports Server (NTRS)

The properties of heteroepitaxial InP cells were determined both before and after 10-MeV proton irradiations. Numerical values, obtained for the diffusion and recombination components of the reverse saturation currents, were found to be consistent with the distribution of dislocations. The radiation resistance of the heteroepitaxial cells was significantly greater than that observed for n/p homoepitaxial InP cells. The carrier removal rate, obtained by C-V measurements, was 1800/cm for 10-MeV protons compared with 2.2/cm for 1-MeV electrons. The high carrier removal rate was found to have no significant effect on the cell's series resistance. It was concluded that the heteroepitaxial cell performance is dominated by the high dislocation density attributable to lattice constant mismatch. Although the efficiencies of the present cells are low, the recent achievement of 13.7 percent AM0 efficiencies using a GaAs substrate demonstrates the marked improvement that can be attained using more appropriate transition layers.

Weinberg, I.; Swartz, C. K.; Brinker, D. J.; Wilt, D. M.

1990-01-01

284

Does vector-free gravity simulate microgravity? Functional and morphologic attributes of clinorotated nerve and muscle grown in cell culture  

NASA Technical Reports Server (NTRS)

Cocultured Xenopus neurons and myocytes were subjected to non-vectorial gravity by clinostat rotation to determine if microgravity, during space flights, may affect cell development and communications. Clinorotated cells showed changes consistent with the hypothesis that cell differentiation, in microgravity, is altered by interference with cytoskeleton-related mechanisms. We found: increases in the myocyte and its nuclear area, "fragmentation" of nucleoli, appearance of neuritic "aneurysms", decreased growth in the presence of "trophic" factors, and decreased yolk utilization. The effects were most notable at 1-10 rpm and depended on the onset and duration of rotation. Some parameters returned to near control values within 48 hrs after cessation of rotation. Cells from cultures rotated at higher speeds (>50 rpm) appeared comparable to controls. Compensation by centrifugal forces may account for this finding. Our data are consistent, in principle, with effects on other, flighted cells and suggest that "vector-free" gravity may simulate certain aspects of microgravity. The distribution of acetylcholine receptor aggregates, on myocytes, was also altered. This indicates that brain development, in microgravity, may also be affected.

Gruener, R.; Hoeger, G.

1988-01-01

285

Thin, high quality GaInP compositionally graded buffer layers grown at high growth rates for metamorphic III–V solar cell applications  

NASA Astrophysics Data System (ADS)

The metamorphic growth of lattice-mismatched materials has allowed optimizing the bandgap combination in multijunction solar cells for the solar spectrum under consideration. Buffer structures are used to accommodate the lattice-mismatch by introducing dislocations and relaxing the material in a controlled way. However, the metamorphic buffers typically involve significant growth time and material usage, which increases the cost of these solar cells. In this work, the thinning of buffer structures with continuously, linearly graded misfit is addressed with the goal of increasing the cost-effectiveness of metamorphic multijunction solar cells. The relaxation dynamics and quality of the buffer layers analyzed were assessed by in-situ stress measurements and ex-situ measurements of residual strain, threading dislocation density and surface roughness. Their ultimate quality has been tested using these buffers as templates for the growth of 1 eV Ga0.73In0.27As solar cells. The deleterious effect of thinning the grade layer of these buffer structures from 2 to 1 ?m was investigated. It is shown that prompting the relaxation of the buffer by using a stepwise misfit jump at the beginning of the grade layer improves the quality of the thinned buffer structure. The residual threading dislocation density of the optimized thin buffers, grown at a high growth rate of 7 ?m/h, is 3×106 cm?2, and solar cells on these buffers exhibit near-ideal carrier collection efficiency and a Voc of 0.62 V at 1-sun direct terrestrial spectrum.

Garcia, I.; France, R. M.; Geisz, J. F.; Simon, J.

2014-05-01

286

Changes in phosphatidylinositol metabolism in response to hyperosmotic stress in Daucus carota L. cells grown in suspension culture.  

PubMed Central

Carrot (Daucus carota L.) cells plasmolyzed within 30 s after adding sorbitol to increase the osmotic strength of the medium from 0.2 to 0.4 or 0.6 osmolal. However, there was no significant change in the polyphosphorylated inositol phospholipids or inositol phosphates or in inositol phospholipid metabolism within 30 s of imposing the hyperosmotic stress. Maximum changes in phosphatidylinositol 4-monophosphate (PIP) metabolism were detected at 5 min, at which time the cells appeared to adjust to the change in osmoticum. There was a 30% decrease in [3H]inositol-labeled PIP. The specific activity of enzymes involved in the metabolism of the inositol phospholipids also changed. The plasma membrane phosphatidylinositol (PI) kinase decreased 50% and PIP-phospholipase C (PIP-PLC) increased 60% compared with the control values after 5 min of hyperosmotic stress. The PIP-PLC activity recovered to control levels by 10 min; however, the PI kinase activity remained below the control value, suggesting that the cells had reached a new steady state with regard to PIP biosynthesis. If cells were pretreated with okadaic acid, the protein phosphatase 1 and 2A inhibitor, the differences in enzyme activity resulting from the hyperosmotic stress were no longer evident, suggesting that an okadaic acid-sensitive phosphatase was activated in response to hyperosmotic stress. Our work suggests that, in this system, PIP is not involved in the initial response to hyperosmotic stress but may be involved in the recovery phase.

Cho, M H; Shears, S B; Boss, W F

1993-01-01

287

Biofilm-Grown Burkholderia cepacia Complex Cells Survive Antibiotic Treatment by Avoiding Production of Reactive Oxygen Species  

PubMed Central

The presence of persister cells has been proposed as a factor in biofilm resilience. In the present study we investigated whether persister cells are present in Burkholderia cepacia complex (Bcc) biofilms, what the molecular basis of antimicrobial tolerance in Bcc persisters is, and how persisters can be eradicated from Bcc biofilms. After treatment of Bcc biofilms with high concentrations of various antibiotics often a small subpopulation survived. To investigate the molecular mechanism of tolerance in this subpopulation, Burkholderia cenocepacia biofilms were treated with 1024 µg/ml of tobramycin. Using ROS-specific staining and flow cytometry, we showed that tobramycin increased ROS production in treated sessile cells. However, approximately 0.1% of all sessile cells survived the treatment. A transcriptome analysis showed that several genes from the tricarboxylic acid cycle and genes involved in the electron transport chain were downregulated. In contrast, genes from the glyoxylate shunt were upregulated. These data indicate that protection against ROS is important for the survival of persisters. To confirm this, we determined the number of persisters in biofilms formed by catalase mutants. The persister fraction in ?katA and ?katB biofilms was significantly reduced, confirming the role of ROS detoxification in persister survival. Pretreatment of B. cenocepacia biofilms with itaconate, an inhibitor of isocitrate lyase (ICL), the first enzyme in the glyoxylate shunt, reduced the persister fraction approx. 10-fold when the biofilms were subsequently treated with tobramycin. In conclusion, most Bcc biofilms contain a significant fraction of persisters that survive treatment with high doses of tobramycin. The surviving persister cells downregulate the TCA cycle to avoid production of ROS and at the same time activate an alternative pathway, the glyoxylate shunt. This pathway may present a novel target for combination therapy.

Van Acker, Heleen; Sass, Andrea; Bazzini, Silvia; De Roy, Karen; Udine, Claudia; Messiaen, Thomas; Riccardi, Giovanna; Boon, Nico; Nelis, Hans J.; Mahenthiralingam, Eshwar; Coenye, Tom

2013-01-01

288

Does vector-free gravity simulate microgravity? Functional and morphologic attributes of clinorotated nerve and muscle grown in cell culture  

NASA Technical Reports Server (NTRS)

Cocultured Xenopus neurons and myocytes were subjected to nonvectorial gravity by clinostat rotation to determine the effects of microgravity on cell development and communications. Observed effects included increases in the myocyte and its nuclear area, fragmentation of nucleoli, the appearance of neuritic aneurysms, decreased growth in the presence of trophic factors, and decreased yolk utilization. These effects were most notable at 1-10 rpm and depended on the onset and duration of rotation. It is found that, in microgravity, cell differentiation is altered by interference with cytoskeleton-related mechanisms. It is suggested that the alteration of the distribution of acetylcholine receptor aggregates on myocytes which occurs might indicate that microgravity affects brain development.

Gruener, Raphael; Hoeger, Glenn

1988-01-01

289

Comparison of MOVPE grown GaAs solar cells using different substrates and group-V precursors  

Microsoft Academic Search

We have investigated the influence of substrate type (GaAs vs. germanium) and of group-V precursor (AsH3 vs. TBAs) on the epitaxial quality of (In)(Al)GaAs layers. We evaluated these layers in terms of morphology, background contamination and doping characteristics. For final benchmarking of the individually optimised processes, we produced p-on-n single junction GaAs solar cells and compared their relative performance. This

J. Derluyn; K. Dessein; G. Flamand; Y. Mols; J. Poortmans; G. Borghs; I. Moerman

2003-01-01

290

Production of single cell protein through fermentation of a perennial grass grown on saline lands with Cellulomonas biazotea  

Microsoft Academic Search

Microbial protein from alkali-treated Leptochloa fusca (kaller grass) was produced by growing Cellulomonasbiazoteain shake flasks and in an aerated 6-l fermentor. Single cell protein, produced in the fermentor contained 56.10 ± 4.64, 60.00 ± 5.04, 11.50 ± 1.34, 12.95 ± 1.24, 3.50 ± 0.24 and 1.00 ± 0.44 true protein, crude protein, crude fibre, ash, cellulose and RNA content respectively.

M. Ibrahim Rajoka

2005-01-01

291

Primary liver cell cultures grown on gas permeable membrane as source for the collection of primary bile  

Microsoft Academic Search

Summary  Isolated rat hepatocytes maintained in primary culture on gas permeable membrane for 20 h form monolayers and establish at\\u000a their cell borders a network of canaliculi (approximate diameter 3.5 ?m). In the presence of the known choleretic bile acid\\u000a dehydrocholate, dilation of canaliculi occurs. When nonfluorescent carboxyfluorescein diacetate ester is added to the culture\\u000a medium, fluorescent carboxyfluorescein appears in the

Ernst Petzinger; Wolfram Föllmann; Helmut Acker; Joachim Hentschel; Karl Zierold; Rolf K. H. Kinne

1988-01-01

292

Effect of pH, aeration and sucrose feeding on the invertase activity of intact S. cerevisiae cells grown in sugarcane blackstrap molasses.  

PubMed

S. cerevisiae was grown in a blackstrap molasses containing medium in batch and fed-batch cultures. The following parameters were varied: pH (from 4.0 to 6.5), dissolved oxygen (DO) (from 0 to 5.0 mg O2 L-1) and sucrose feeding rate. When glucose concentration (S) was higher than 0.5 g L-1 a reduction in the specific invertase activity of intact cells (v) and an oscillatory behavior of v values during fermentation were observed. Both the invertase reduction and the oscillatory behavior of v values could be related to the glucose inhibitory effect on invertase biosynthesis. The best culture conditions for attaining S. cerevisiae cells suitable for invertase production were: temperature = 30 degrees C; pH = 5.0; DO = 3.3 mg O2 L-1; (S) = 0.5 g L-1 and sucrose added into the fermenter according to the equations: (V-Vo) = t2/16 or (V - Vo) = (Vf - Vo).(e0.6t-1)/10. PMID:7576463

Vitolo, M; Duranti, M A; Pellegrim, M B

1995-08-01

293

Reflectivity and topography of cells grown on glass-coverslips measured with phase-shifted laser feedback interference microscopy.  

PubMed

In spite of the advantages associated with the molecular specificity of fluorescence imaging, there is still a significant need to augment these approaches with label-free imaging. Therefore, we have implemented a form of interference microscopy based upon phase-shifted, laser-feedback interferometry and developed an algorithm that can be used to separate the contribution of the elastically scattered light by sub-cellular structures from the reflection at the coverslip-buffer interface. The method offers an opportunity to probe protein aggregation, index of refraction variations and structure. We measure the topography and reflection from calibration spheres and from stress fibers and adhesions in both fixed and motile cells. Unlike the data acquired with reflection interference contrast microscopy, where the reflection from adhesions can appear dark, our approach demonstrates that these regions have high reflectivity. The data acquired from fixed and live cells show the presence of a dense actin layer located ? 100 nm above the coverslip interface. Finally, the measured dynamics of filopodia and the lamella in a live cell supports retrograde flow as the dominate mechanism responsible for filopodia retraction. PMID:21833378

At?lgan, Erdinç; Ovryn, Ben

2011-08-01

294

Reflectivity and topography of cells grown on glass-coverslips measured with phase-shifted laser feedback interference microscopy  

PubMed Central

In spite of the advantages associated with the molecular specificity of fluorescence imaging, there is still a significant need to augment these approaches with label-free imaging. Therefore, we have implemented a form of interference microscopy based upon phase-shifted, laser-feedback interferometry and developed an algorithm that can be used to separate the contribution of the elastically scattered light by sub-cellular structures from the reflection at the coverslip-buffer interface. The method offers an opportunity to probe protein aggregation, index of refraction variations and structure. We measure the topography and reflection from calibration spheres and from stress fibers and adhesions in both fixed and motile cells. Unlike the data acquired with reflection interference contrast microscopy, where the reflection from adhesions can appear dark, our approach demonstrates that these regions have high reflectivity. The data acquired from fixed and live cells show the presence of a dense actin layer located ? 100 nm above the coverslip interface. Finally, the measured dynamics of filopodia and the lamella in a live cell supports retrograde flow as the dominate mechanism responsible for filopodia retraction.

At?lgan, Erdinc; Ovryn, Ben

2011-01-01

295

The in vivo positional identity gene expression code is not preserved in neural stem cells grown in culture.  

PubMed

Neural stem cell specification depends on antero-posterior (AP) and dorso-ventral (DV) information provided during development. In the present study we identified similar neural stem cell (NSC) populations along the AP axis of the mouse central nervous system: the 'early' NSCs responsive to fibroblast growth factor-2 and the 'late' NSCs responsive to epidermal growth factor (EGF). Gene expression analysis shows that AP and DV transcription factor code is not preserved in NSCs in culture. Neurospheres generated with EGF from different regions showed Emx2, En2 and Krox20 expression beyond their corresponding AP restricted areas (telencephalon, mesencephalon and rhomboencephalon, respectively). Hox genes were rarely expressed. DV markers such as Pax7 and Dbx1 were not expressed in neurosphere cells, whereas Pax6 and Nkx2.1 were highly expressed independently of the NSC source region. In general, this pattern was found under different culture conditions. We propose that signals surrounding NSCs determine their positional identity gene expression code, which may be relevant to establish their definitive fate. PMID:12956707

Santa-Olalla, Jesús; Baizabal, José-Manuel; Fregoso, Mariana; del Carmen Cárdenas, María; Covarrubias, Luis

2003-09-01

296

Bone and cartilage tissue constructs grown using human bone marrow stromal cells, silk scaffolds and rotating bioreactors.  

PubMed

Human bone marrow contains a population of bone marrow stromal cells (hBMSCs) capable of forming several types of mesenchymal tissues, including bone and cartilage. The present study was designed to test whether large cartilaginous and bone-like tissue constructs can be selectively engineered using the same cell population (hBMSCs), the same scaffold type (porous silk) and same hydrodynamic environment (construct settling in rotating bioreactors), by varying the medium composition (chondrogenic vs. osteogenic differentiation factors). The hBMSCs were harvested, expanded and characterized with respect to their differentiation potential and population distribution. Passage two cells were seeded on scaffolds and cultured for 5 weeks in bioreactors using osteogenic, chondrogenic or control medium. The three media yielded constructs with comparable wet weights and compressive moduli ( approximately 25 kPa). Chondrogenic medium yielded constructs with higher amounts of DNA (1.5-fold) and glycosaminoglycans (GAG, 4-fold) per unit wet weight (ww) than control medium. In contrast, osteogenic medium yielded constructs with higher dry weight (1.6-fold), alkaline phosphatase (AP) activity (8-fold) and calcium content (100-fold) per unit ww than control medium. Chondrogenic medium yielded constructs that were weakly positive for GAG by contrast-enhanced MRI and alcian blue stain, whereas osteogenic medium yielded constructs that were highly mineralized by microCT and von Kossa stain. Engineered bone constructs were large (8mm diameter x 2mm thick disks) and resembled trabecular bone with respect to structure and mineralized tissue volume fraction (12%). PMID:16895736

Marolt, Darja; Augst, Alexander; Freed, Lisa E; Vepari, Charu; Fajardo, Robert; Patel, Nipun; Gray, Martha; Farley, Michelle; Kaplan, David; Vunjak-Novakovic, Gordana

2006-12-01

297

Novel antigens expressed by Aeromonas salmonicida grown in vivo.  

PubMed Central

Virulent and avirulent Aeromonas salmonicida strains grown inside intraperitoneal implants in Rainbow trout (Oncorhynchus mykiss) were examined for unique antigen expression. Western blots (immunoblots), performed with immune rabbit serum raised against in vivo-grown cells, revealed several unique antigens. With the exception of lipopolysaccharide (LPS), these novel antigens were destroyed after proteinase K treatment. The majority of these antigens were not induced in vitro in response to either iron limitation or anaerobiosis. In addition, electron microscopy demonstrated the presence of a putative capsule on in vivo-grown cells. Purification and fractionation of this carbohydrate material from cells grown in carbon-rich synthetic media resulted in the isolation and separation of an antigenically distinct LPS not seen with cells grown in standard media. Antiserum raised against in vivo-grown cells recognized both this LPS and the typical LPS of A. salmonicida apparent in in vitro-grown cells. Antiserum raised against in vitro-grown cells recognized only the LPS expressed in vitro. Antiserum directed against in vivo-grown cells was approximately 10 times more sensitive than serum directed against in vitro-grown cells in detecting A. salmonicida in infected fish kidney tissue. Images

Thornton, J C; Garduno, R A; Carlos, S J; Kay, W W

1993-01-01

298

Impact of trichloroethylene contaminated groundwater discharged to the main canal and Indian River lagoon, Vero Beach, Florida  

SciTech Connect

Groundwater highly contaminated with trichloroethylene (TCE) from a leaky storage tank was detected in Vero Beach, Florida in 1978. Aware of this problem, the local and state authorities gave permission to pump out the contaminated water as a means of reducing concentrations in the aquifer. The water was air sprayed to strip the organic compounds and subsequently discharged and mixed by means of a hydraulic pump in the drainage canal. The average discharge rate of contaminated water into the canal was approximately 0.2 million gallons per day. This project was initiated to determine the spatial distribution of pollutants in the canal and river as well as rainfall and canal flow rate effects on water, sediment, and biological organisms. Prior to flushing the well, a baseline survey of trichloroethylene and other related compounds in the canal and river was performed.

Wang, T.; Lenahan, R.; Kanik, M.

1985-04-01

299

Antioxidant Response to NaCl Stress in a Control and an NaCI-Tolerant Cotton Cell Line Grown in the Presence of Paraquat, Buthionine Sulfoximine, and Exogenous Glutathione  

Microsoft Academic Search

A cotton (Gossypium hirsutum 1.) control and NaCI-tolerant cell line (cv Coker 312) were grown on media with or without NaCl in the presence or absence of paraquat, buthionine sulfoximine, and oxidized glutathione. On medium with 150 mM NaCl the NaCI-tolerant cell line exhibited no reduction in growth, whereas a 96% reduction was observed in the control lhe. The NaCI-tolerant

Dalton R. Cossett; Stephen W. Banks; Eddie P. Millhollon; M. Cran Lucas

300

Evaluation of defects generation in crystalline silicon ingot grown by cast technique with seed crystal for solar cells  

PubMed Central

Although crystalline silicon is widely used as substrate material for solar cell, many defects occur during crystal growth. In this study, the generation of crystalline defects in silicon substrates was evaluated. The distributions of small-angle grain boundaries were observed in substrates sliced parallel to the growth direction. Many precipitates consisting of light elemental impurities and small-angle grain boundaries were confirmed to propagate. The precipitates mainly consisted of Si, C, and N atoms. The small-angle grain boundaries were distributed after the precipitation density increased. Then, precipitates appeared at the small-angle grain boundaries. We consider that the origin of the small-angle grain boundaries was lattice mismatch and/or strain caused by the high-density precipitation.

Tachibana, Tomihisa; Sameshima, Takashi; Kojima, Takuto; Arafune, Koji; Kakimoto, Koichi; Miyamura, Yoshiji; Harada, Hirofumi; Sekiguchi, Takashi; Ohshita, Yoshio; Ogura, Atsushi

2012-01-01

301

Inhibition of Vimentin or ?1-integrin Reverts Morphology of Prostate Tumor Cells Grown in Laminin-rich ECM gels and Reduces Tumor Growth in vivo  

PubMed Central

Prostate epithelial cells grown embedded in laminin-rich extracellular matrix (lrECM) undergo morphological changes that closely resemble their architecture in vivo. In this study, growth characteristics of three human prostate epithelial sublines derived from the same cellular lineage, but displaying different tumorigenic and metastatic properties in vivo, were assessed in three-dimensional (3D) lrECM gels. M12, a highly tumorigenic and metastatic subline, was derived from the immortalized, prostate epithelial P69 cell line by selection in athymic, nude mice and found to contain a deletion of 19p-q13.1. The stable re-introduction of an intact human chromosome 19 into M12 resulted in a poorly tumorigenic subline, designated F6. When embedded in lrECM gels, the parental, non-tumorigenic P69 line produced acini with clearly defined lumena. Immunostaining with antibodies to ?-catenin, E-cadherin, or ?6- and ?1-integrins showed polarization typical of glandular epithelium. In contrast, the metastatic M12 subline produced highly disorganized cells with no evidence of polarization. The F6 subline reverted to acini-like structures exhibiting basal polarity marked with integrins. Reducing either vimentin levels via siRNA interference or the expression of ?6-, ?1-integrins by the addition of blocking antibodies, reorganized the M12 subline into forming polarized acini. The loss of vimentin significantly reduced M12-Vim tumor growth when assessed by subcutaneous injection in athymic mice. Thus, tumorigenicity in vivo correlated with disorganized growth in 3D lrECM gels. These studies suggest that the levels of vimentin and ?1-integrin play a key role in the homeostasis of the normal acinus in prostate and that their dysregulation may lead to tumorigenesis.

Zhang, Xueping; Fournier, Marcia V.; Ware, Joy L.; Bissell, Mina J.; Jacoub, Adly; Zehner, Zendra E.

2009-01-01

302

Regulation of Cell Division, Biofilm Formation, and Virulence by FlhC in Escherichia coli O157:H7 Grown on Meat?†  

PubMed Central

To understand the continuous problems that Escherichia coli O157:H7 causes as food pathogen, this study assessed global gene regulation in bacteria growing on meat. Since FlhD/FlhC of E. coli K-12 laboratory strains was previously established as a major control point in transducing signals from the environment to several cellular processes, this study compared the expression pattern of an E. coli O157:H7 parent strain to that of its isogenic flhC mutant. This was done with bacteria that had been grown on meat. Microarray experiments revealed 287 putative targets of FlhC. Real-time PCR was performed as an alternative estimate of transcription and confirmed microarray data for 13 out of 15 genes tested (87%). The confirmed genes are representative of cellular functions, such as central metabolism, cell division, biofilm formation, and pathogenicity. An additional 13 genes from the same cellular functions that had not been hypothesized as being regulated by FlhC by the microarray experiment were tested with real-time PCR and also exhibited higher expression levels in the flhC mutant than in the parent strain. Physiological experiments were performed and confirmed that FlhC reduced the cell division rate, the amount of biofilm biomass, and pathogenicity in a chicken embryo lethality model. Altogether, this study provides valuable insight into the complex regulatory network of the pathogen that enables its survival under various environmental conditions. This information may be used to develop strategies that could be used to reduce the number of cells or pathogenicity of E. coli O157:H7 on meat by interfering with the signal transduction pathways.

Sule, Preeti; Horne, Shelley M.; Logue, Catherine M.; Pruss, Birgit M.

2011-01-01

303

Metabolic Regulation of "Ca. Methylacidiphilum Fumariolicum" SolV Cells Grown Under Different Nitrogen and Oxygen Limitations  

PubMed Central

Aerobic methanotrophic bacteria can use methane as their sole energy source. The discovery of “Ca. Methylacidiphilum fumariolicum” strain SolV and other verrucomicrobial methanotrophs has revealed that the ability of bacteria to oxidize CH4 is much more diverse than has previously been assumed in terms of ecology, phylogeny, and physiology. A remarkable characteristic of the methane-oxidizing Verrucomicrobia is their extremely acidophilic phenotype, growing even below pH 1. In this study we used RNA-Seq to analyze the metabolic regulation of “Ca. M. fumariolicum” SolV cells growing at ?max in batch culture or under nitrogen fixing or oxygen limited conditions in chemostats, all at pH 2. The analysis showed that two of the three pmoCAB operons each encoding particulate methane monoxygenases were differentially expressed, probably regulated by the available oxygen. The hydrogen produced during N2 fixation is apparently recycled as demonstrated by the upregulation of the genes encoding a Ni/Fe-dependent hydrogenase. These hydrogenase genes were also upregulated under low oxygen conditions. Handling of nitrosative stress was shown by the expression of the nitric oxide reductase encoding genes norB and norC under all conditions tested, the upregulation of nitrite reductase nirK under oxygen limitation and of hydroxylamine oxidoreductase hao in the presence of ammonium. Unraveling the gene regulation of carbon and nitrogen metabolism helps to understand the underlying physiological adaptations of strain SolV in view of the harsh conditions of its natural ecosystem.

Khadem, Ahmad F.; Pol, Arjan; Wieczorek, Adam S.; Jetten, Mike S. M.; Op den Camp, Huub J. M.

2012-01-01

304

Video of Tissue Grown in Space in NASA Bioreactor  

NASA Technical Reports Server (NTRS)

Principal investigator Leland Chung grew prostate cancer and bone stromal cells aboard the Space Shuttle Columbia during the STS-107 mission. Although the experiment samples were lost along with the ill-fated spacecraft and crew, he did obtain downlinked video of the experiment that indicates the enormous potential of growing tissues in microgravity. Cells grown aboard Columbia had grown far larger tissue aggregates at day 5 than did the cells grown in a NASA bioreactor on the ground.

2003-01-01

305

Comparative immunogenicity and cross-clade protective efficacy of mammalian cell-grown inactivated and live attenuated H5N1 reassortant vaccines in ferrets.  

PubMed

Continued H5N1 virus infection in humans highlights the need for vaccine strategies that provide cross-clade protection against this rapidly evolving virus. We report a comparative evaluation in ferrets of the immunogenicity and cross-protective efficacy of isogenic mammalian cell-grown, live attenuated influenza vaccine (LAIV) and adjuvanted, whole-virus, inactivated influenza vaccine (IIV), produced from a clade 1 H5N1 6:2 reassortant vaccine candidate (caVN1203-Len17rg) based on the cold-adapted A/Leningrad/134/17/57 (H2N2) master donor virus. Two doses of LAIV or IIV provided complete protection against lethal homologous H5N1 virus challenge and a reduction in virus shedding and disease severity after heterologous clade 2.2.1 H5N1 virus challenge and increased virus-specific serum and nasal wash antibody levels. Although both vaccines demonstrated cross-protective efficacy, LAIV induced higher levels of nasal wash IgA and reduction of heterologous virus shedding, compared with IIV. Thus, enhanced respiratory tract antibody responses elicited by LAIV were associated with improved cross-clade protection. PMID:21957153

Gustin, Kortney M; Maines, Taronna R; Belser, Jessica A; van Hoeven, Neal; Lu, Xuihua; Dong, Libo; Isakova-Sivak, Irina; Chen, Li-Mei; Voeten, J Theo M; Heldens, Jacco G M; van den Bosch, Han; Cox, Nancy J; Tumpey, Terrence M; Klimov, Alexander I; Rudenko, Larisa; Donis, Ruben O; Katz, Jacqueline M

2011-11-15

306

Optical and electrical characterization of CdS-Glycine thin films with ammonia free buffer grown at different temperatures for solar cells applications  

NASA Astrophysics Data System (ADS)

In this work we report the fabrication and electro-optical characterization of CdS thin films using glycine as complexing agent with ammonia and ammonia free buffer by the Chemical Bath Deposition (CBD) method. The CdS thin films were grown at different temperatures of 50, 60, 70 and 80 °C in a thermal water bath. The morphology of these films was determined using atomic force microscopy; the resultant films were homogeneous, well adhered to the substrate, and specularly reflecting with a varying color depending on the deposition temperature. Transmittance and reflectance measurements of thermally treated CdS films were carried to study the effect of the ammonia buffer on its optical properties and bandgap. The crystallinity of the CdS thin films was determined by means of X Ray diffraction measurements. Therefore, for this study, an ammonia-free complexing agent has been taken for the deposition of CdS. Among different methods, which are being used for the preparation of CdS films, Chemical Bath Deposition (CBD) is the most attractive due to its low cost, easy to handle and large possibilities regarding doping and deposition on various substrates. In particular it can be used to easily obtain field effect devices by depositing CdS thin films over a SiO2/Si substrate. Heterostructures with interesting physical properties can be imagined, realized and tested in this way.. Structures CdS/PbS also were realized and have shown good solar cell characteristics.

Berman-Mendoza, D.; Quiñones-Urías, D.; Ferra-González, S.; Vera-Marquina, A.; Rojas-Hernández, A.; Gómez Fuentes, R.; García-Juárez, A.; Leal-Cruz, A. L.; Ramos-Carrasco, A.

2013-11-01

307

Metabolic Regulation of "Ca. Methylacidiphilum Fumariolicum" SolV Cells Grown Under Different Nitrogen and Oxygen Limitations.  

PubMed

Aerobic methanotrophic bacteria can use methane as their sole energy source. The discovery of "Ca. Methylacidiphilum fumariolicum" strain SolV and other verrucomicrobial methanotrophs has revealed that the ability of bacteria to oxidize CH(4) is much more diverse than has previously been assumed in terms of ecology, phylogeny, and physiology. A remarkable characteristic of the methane-oxidizing Verrucomicrobia is their extremely acidophilic phenotype, growing even below pH 1. In this study we used RNA-Seq to analyze the metabolic regulation of "Ca. M. fumariolicum" SolV cells growing at ?(max) in batch culture or under nitrogen fixing or oxygen limited conditions in chemostats, all at pH 2. The analysis showed that two of the three pmoCAB operons each encoding particulate methane monoxygenases were differentially expressed, probably regulated by the available oxygen. The hydrogen produced during N(2) fixation is apparently recycled as demonstrated by the upregulation of the genes encoding a Ni/Fe-dependent hydrogenase. These hydrogenase genes were also upregulated under low oxygen conditions. Handling of nitrosative stress was shown by the expression of the nitric oxide reductase encoding genes norB and norC under all conditions tested, the upregulation of nitrite reductase nirK under oxygen limitation and of hydroxylamine oxidoreductase hao in the presence of ammonium. Unraveling the gene regulation of carbon and nitrogen metabolism helps to understand the underlying physiological adaptations of strain SolV in view of the harsh conditions of its natural ecosystem. PMID:22848206

Khadem, Ahmad F; Pol, Arjan; Wieczorek, Adam S; Jetten, Mike S M; Op den Camp, Huub J M

2012-01-01

308

Cordyceps militaris Grown on Germinated Soybean Induces G2/M Cell Cycle Arrest through Downregulation of Cyclin B1 and Cdc25c in Human Colon Cancer HT-29 Cells.  

PubMed

Cordyceps militaris (CM) is an insect-borne fungus that has been used in traditional Chinese medicine because of its wide range of pharmacological activities. In this paper, we studied CM grown on germinated soybean (GSC) and investigated the possible mechanisms underlying antiproliferative effect of GSC on HT-29 human colon cancer cells. In comparison with CM extracts and germinated soybean (GS) BuOH extracts, BuOH extracts of GSC showed remarkable inhibitory and antiproliferative effects on HT-29 colon cancer cells. After GSC treatment, HT-29 cells became smaller and irregular in shape. High G2/M phase cell populations were observed in the GSC-treated group. The levels of cyclin B1 and Cdc25 in the GSC-treated group were lower than those in the control group. These findings suggest that GSC BuOH extracts might act as an effective anti-proliferative agent by inducing G2/M cell cycle arrest in colon cancer cells. PMID:22474493

Mollah, Mohammad Lalmoddin; Park, Dong Ki; Park, Hye-Jin

2012-01-01

309

Cordyceps militaris Grown on Germinated Soybean Induces G2/M Cell Cycle Arrest through Downregulation of Cyclin B1 and Cdc25c in Human Colon Cancer HT-29 Cells  

PubMed Central

Cordyceps militaris (CM) is an insect-borne fungus that has been used in traditional Chinese medicine because of its wide range of pharmacological activities. In this paper, we studied CM grown on germinated soybean (GSC) and investigated the possible mechanisms underlying antiproliferative effect of GSC on HT-29 human colon cancer cells. In comparison with CM extracts and germinated soybean (GS) BuOH extracts, BuOH extracts of GSC showed remarkable inhibitory and antiproliferative effects on HT-29 colon cancer cells. After GSC treatment, HT-29 cells became smaller and irregular in shape. High G2/M phase cell populations were observed in the GSC-treated group. The levels of cyclin B1 and Cdc25 in the GSC-treated group were lower than those in the control group. These findings suggest that GSC BuOH extracts might act as an effective anti-proliferative agent by inducing G2/M cell cycle arrest in colon cancer cells.

Mollah, Mohammad Lalmoddin; Park, Dong Ki; Park, Hye-Jin

2012-01-01

310

Expression of CXC chemokines and their receptors is modulated during chondrogenic differentiation of human mesenchymal stem cells grown in three-dimensional scaffold: evidence in native cartilage.  

PubMed

Chemokines contribute to the maintenance of cartilage homeostasis. To evaluate the role of CXC chemokines CXCL8 (interleukin-8), CXCL10 (interferon-gamma-inducible protein-10), CXCL12 (stroma-derived factor-1) and CXCL13 (B-cell attracting chemokine-1) and their receptors, respectively CXCR1-2, CXCR3, CXCR4, and CXCR5, during chondrogenic differentiation of human mesenchymal stromal cells (h-MSCs), we used a well-defined in vitro model. Chondrogenic differentiation was analyzed on h-MSCs grown on hyaluronic acid-based biomaterial in the presence or absence of transforming growth factor-beta, and the expression and modulation of CXC chemokines and receptors were evaluated at different time points. Real-time polymerase chain reaction was performed to analyze their expression at the messenger ribonucleic acid (mRNA) level, and immunohistochemistry and enzyme-linked immunosorbent assay were used to evaluate their expression at the protein level. Human articular cartilage biopsies were used to evaluate chemokine and receptor expression in normal tissue. We found no expression of CXCR1, CXCR2, CXCR3, or CXCL10 at the mRNA level. CXCL8 mRNA was down-modulated, whereas at the protein level, we found greater release of this chemokine. CXCR4 and its ligand CXCL12 were down-modulated during chondrogenesis. By contrast, CXCR5 was up-regulated, whereas its ligand CXCL13 was lower. These data were also confirmed on human articular cartilage. These findings show that, during in vitro h-MSC chondrogenic differentiation, chemokine and receptor expression was specifically induced or repressed. This was in line with what the authors also found in normal articular cartilage, suggesting a role in differentiation and maturation of a cartilage-like structure in vitro and consequently the regulation of cartilage homeostasis. PMID:18333808

Cristino, Sandra; Piacentini, Anna; Manferdini, Cristina; Codeluppi, Katia; Grassi, Francesco; Facchini, Andrea; Lisignoli, Gina

2008-01-01

311

Aeromonas salmonicida grown in vivo.  

PubMed Central

The virulent fish pathogen Aeromonas salmonicida was rapidly killed in vivo when restricted inside a diffusion chamber implanted intraperitoneally in rainbow trout. After a period of regrowth, the survivors had acquired resistance to host-mediated bacteriolysis, phagocytosis, and oxidative killing, properties which were subsequently lost by growth in vitro. Resistance to bacteriolysis and phagocytosis was associated with a newly acquired capsular layer revealed by acidic polysaccharide staining and electron microscopy. This capsular layer shielded the underlying, regular surface array (S-layer) from immunogold labeling with a primary antibody to the S-layer protein. Resistance to oxidative killing was mediated by a mechanism not associated with the presence of the capsular layer. An attenuated vaccine strain of A. salmonicida grown in vivo failed to express the capsular layer. Consequently, the in vivo-grown cells of this attenuated strain remained as sensitive to bacteriolysis, and as avidly adherent to macrophages, as the in vitro-grown cells. The importance of these new virulence determinants and their relation to the known virulence factors of A. salmonicida are discussed. Images

Garduno, R A; Thornton, J C; Kay, W W

1993-01-01

312

Pathogenic Trichomonas vaginalis cytotoxicity to cell culture monolayers  

Microsoft Academic Search

Exposure of monolayer cultures of human urogenital and vaginal (HeLa), human epithelial (HEp-2), normal baboon testicular (NBT), and monkey kidney (Vero) cells to live pathogenic Trichomonas vaginalis resulted in extensive disruption of monolayers. Trypan blue was taken up by all host cells released from cell monolayers, which indicated irreversible damage of these cell types by trichomonads. Time and dose related

J F Alderete; E Pearlman

1984-01-01

313

Carrier dynamics in bulk 1eV InGaAsNSb materials and epitaxial lift off GaAs-InAlGaP layers grown by MOVPE for multi-junction solar cells  

NASA Astrophysics Data System (ADS)

III-V multi-junction solar cells are based on a triple-junction design that consists of an InGaP top junction, a GaAs middle junction, and a bottom junction that employs either a 1eV material grown on the GaAs substrate or InGaAs grown on the Ge substrate. The most promising 1 eV material that is currently under extensive investigation is bulk dilute nitride such as InGaAsN(Sb) lattice matched to GaAs substrates. Both approaches utilizing dilute nitrides and lattice-mismatched InGaAs layers have a potential to achieve high performance triple-junction solar cells. In addition, it will be beneficial for both commercial and space applications if III-V triple-junction solar cells can significantly reduce weight and can be manufactured cost effectively while maintaining high efficiency. The most attractive approach to achieve these goals is to employ full-wafer epitaxial lift off (ELO) technology, which can eliminate the substrate weight and also enable multiple substrate re-usages. For the present study, we employed time-resolved photoluminescence (TR-PL) techniques to study carrier dynamics in MOVPE-grown bulk dilute nitride layers lattice matched to GaAs substrates, where carrier lifetime measurements are crucial in optimizing MOVPE materials growth. We studied carrier dynamics in InGaAsN(Sb) layers with different amounts of N incorporated. Carrier lifetimes were also measured from InGaAsN(Sb) layers at different stages of post-growth thermal annealing steps. Post-growth annealing yielded significant improvements in carrier lifetimes of InGaAsNSb double hetero-structure (DH) samples compared to InGaAsN DH samples possibly due to the surfactant effect of Sb. In addition, we studied carrier dynamics in MOVPE-grown GaAs-InAl(Ga)P layers grown on GaAs substrates. The structures were grown on top of a thin AlAs release layer, which allowed epitaxial layers grown on top of the AlAs layer to be removed from the substrate. The GaAs layers had various doping densities and thicknesses. We present our TR-PL results from both pre- and post-ELO processed GaAs-InAl(Ga)P samples.

Sin, Yongkun; LaLumondiere, Stephen; Lotshaw, William; Moss, Steven C.; Kim, Tae Wan; Forghani, Kamran; Mawst, Luke J.; Kuech, Thomas F.; Tatavarti, Rao; Wibowo, Andree; Pan, Noren

2013-03-01

314

Electrical and optical properties of Si-doped InP grown by solid source molecular beam epitaxy using a valved phosphorus cracker cell  

NASA Astrophysics Data System (ADS)

We report on the electrical and optical properties of silicon (Si)-doped InP layers grown by solid-source molecular beam epitaxy using a valved phosphorus cracker cell. Within the range of Si effusion cell temperatures investigated (900-1200 °C), the highest electron concentration obtained was 1.1×1020 cm-3. A saturation phenomenon was observed for the electron concentration at higher Si cell temperatures. 300 and 77 K Hall mobility data were used to determine the compensation ratios by comparing them with the theoretical data. Although the Hall data show that the compensation ratio increases with the increase in carrier concentration, the exact values are not certain because the theoretical calculation overestimates the mobility values at higher carrier concentrations. The saturation phenomenon of electron concentration in InP may be considered due to the Si atoms occupying both the In and P lattice sites, or Si donors located at the interstitial sites. The 300 K Hall mobility and the concentration data measured were found to fit the Hilsum expression well. The mobility values obtained in this study are better than or comparable to reported data in the past, indicating good material quality. 5 K photoluminescence (PL) measurements showed two peaks for the undoped and low doped InP layers corresponding to the neutral donor-bound exciton transitions (D0-X) and the acceptor-related transitions (D-A), respectively. When the doping level was increased, the near-band edge (D0-X) recombination peak becomes broadened and asymmetric due to changes in the donor level density of states and relaxation of the wave vector conservation rule. The full-width at half-maximum (FWHM) value of the PL peak position increased when the doping concentration was increased. An empirical equation was developed to fit this variation, which provides a convenient way of determining the dopant concentration from the experimental FWHM value. The near-band edge peak positions shifted to higher energy with the increase of doping level due to the band filling effect. This shift agreed well with the calculations based on the Burstein-Moss shift and the band gap narrowing effect considering a nonparabolic conduction band.

Zheng, H. Q.; Radahakrishnan, K.; Yoon, S. F.; Ng, G. I.

2000-06-01

315

Rickettsia prowazekii requires host cell serine and glycine for growth.  

PubMed Central

The growth requirement of Rickettsia prowazekii for the amino acids serine and glycine was assessed in both wild-type cell lines and a mutant cell line. X-irradiated L929 cells supported the growth of R. prowazekii when the cells were incubated in Eagle minimal essential medium supplemented with serum. In contrast, in this medium, X-irradiated Vero cells did not support the growth of rickettsiae unless cycloheximide, serine, or glycine was added. Other nonessential amino acids, additional glucose, and potential products of host cell metabolism of serine and glycine were nonstimulatory. The concentration of serine or glycine required to support rickettsial growth had no effect on the doubling time of uninfected, unirradiated Vero cells. A comparison of intracellular amino acid pools indicated that the serine and glycine concentrations in mock-infected Vero cells were approximately 31 and 14% of the respective concentrations in mock-infected L929 cells. The pools of both amino acids in Vero cells increased markedly upon treatment of the cells with cycloheximide. Interconversion of serine and glycine catalyzed by serine hydroxymethyltransferase was detected in cell-free extracts of purified rickettsiae. However, this enzymatic activity did not permit rickettsial growth in a glycine-requiring clone (772-56d) of the Chinese hamster ovary cell CHO-K1 in the absence of glycine supplementation. These data indicate that R. prowazekii depends on the host cell for serine or glycine.

Austin, F E; Turco, J; Winkler, H H

1987-01-01

316

Effects of substrate temperature on the properties of In 0.48Ga 0.52P grown by molecular-beam epitaxy using a valved phosphorus cracker cell  

NASA Astrophysics Data System (ADS)

We report the molecular-beam epitaxial (MBE) growth of high-quality In 0.48Ga 0.52P epitaxial layers on GaAs substrate using a valved phosphorus cracker cell at a wide range of substrate temperature ( Ts) from 440 to 520°C. Film characterization was carried out using double-axis X-ray diffraction (XRD), low-temperature photoluminescence (PL) and Hall-effect measurement. Typical InGaP layers showed a lattice mismatch of <10 -3 and PL full-width at half-maximum (FWHM) as low as 7 meV at 10 K, indicating materials with good structural and optical quality. The lattice mismatch remained relatively constant for samples grown between 440 and 500°C but increased significantly as the substrate temperature was increased to 520°C. Compositional measurements using XRD showed that indium desorption was significant in samples grown at substrate temperatures exceeding 500°C. The PL peak energy decreased from 2.014±0.008 to 1.968±0.008 eV as the substrate temperature was increased from 440 to 520°C. The highest PL peak energy of 2.014±0.008 eV which was indicative of the highest band gap was measured from the sample grown at Ts=440°C. The results suggest that highly disordered (i.e. random) InGaP materials can be grown at low substrate temperature with excellent structural and optical quality.

Yoon, S. F.; Mah, K. W.; Zheng, H. Q.; Zhang, P. H.

1998-08-01

317

Formation of a spherical multicellular aggregate (spheroid) of animal cells in the pores of polyurethane foam as a cell culture substratum and its application to a hybrid artificial liver  

Microsoft Academic Search

Monkey kidney cells (Vero), human embryonic kidney cells (293), human liver cells (PLC\\/PRF\\/5), and primary rat, dog, and porcine hepatocytes formed spherical multicellular aggregates (spheroids) in the pores of polyurethane foam which was used as a cell culture substratum. These spheroids of various cell types express high cell activity for a long period. A practical hybrid artificial liver support system

Hiroyuki Ijima; Kohji Nakazawa; Hiroshi Mizumoto; Taku Matsushita; Kazumori Funatsu

1998-01-01

318

Cell Fusion Activities of Hantaan Virus Envelope Glycoproteins  

PubMed Central

Hantaan virus (HTNV)-infected Vero E6 cells undergo cell fusion with both infected and uninfected cells under low-pH conditions. Flow cytometry and fluorescence microscopy of HTNV-infected Vero E6 cells showed that envelope glycoproteins (GPs) were located both on the cell surface and in the cytoplasm. Neutralizing monoclonal antibodies (MAbs) against the G1 and G2 envelope GPs inhibited cell fusion, whereas nonneutralizing MAbs against G1 or G2 and MAbs against the nucleocapsid protein (NP) did not. Transfected Vero E6 cells that expressed GPs but not those that expressed NP fused and formed syncytia. These results indicate that HTNV GPs act as fusogens at the cell surface. No fusion activity was observed either in infected Vero cells that were passaged more than 150 times or in BHK-21 cells, although GPs appeared to localize to the cell surface. This variability in fusion induction suggests the involvement of host cell factors in the process of cell membrane fusion.

Ogino, Michiko; Yoshimatsu, Kumiko; Ebihara, Hideki; Araki, Koichi; Lee, Byoung-Hee; Okumura, Megumi; Arikawa, Jiro

2004-01-01

319

Investigation of anodic and chemical oxides grown on p-type InP with applications to surface passivation for n(+)-p solar cell fabrication  

NASA Technical Reports Server (NTRS)

Most of the previously reported InP anodic oxides were grown on a n-type InP with applications to fabrication of MISFET structures and were described as a mixture of In2O3 and P2O5 stoichiometric compounds or nonstoichiometric phases which have properties similar to crystalline compounds In(OH)3, InPO4, and In(PO3)3. Details of the compositional change of the anodic oxides grown under different anodization conditions were previously reported. The use of P-rich oxides grown either by anodic or chemical oxidation are investigated for surface passivation of p-type InP and as a protective cap during junction formation by closed-ampoule sulfur diffusion. The investigation is based on but not limited to correlations between PL intensity and X-ray photoelectron spectroscopy (XPS) chemical composition data.

Faur, Maria; Faur, Mircea; Goradia, Manju; Goradia, Chandra; Jenkins, Phillip; Jayne, Douglas; Weinberg, Irving

1991-01-01

320

Application of an immunoaffinity-based preconcentration method for mass spectrometric analysis of the O-chain polysaccharide of Aeromonas salmonicida from in vitro- and in vivo-grown cells.  

PubMed

In this study, application of magnetic beads (Dynabeads) coated with Aeromonas salmonicida lipopolysaccharide-specific polyclonal antisera to MS-based characterization of bacterial lipopolysaccharides has been evaluated. The results showed that the affinity-based preconcentration strategy resulted in at least a 100-fold increase in the detection of sensitivity, affording direct capillary electrophoresis (CE)-MS analysis of A. salmonicida lipopolysaccharide O-chain polysaccharide from in vitro-cultured cells. Subsequent CE-MS analysis of in vivo-grown cells of A. salmonicida confirmed significant changes in the structure of the lipopolysaccharide O-chain polysaccharide as a result of in vivo cultivation. PMID:19456871

Wang, Zhan; Liu, Xin; Garduño, Elizabeth; Garduño, Rafael A; Li, Jianjun; Altman, Eleonora

2009-06-01

321

Investigation of the uptake of drugs by individual cells using a scanning proton microprobe (SPM).  

PubMed

In this paper we demonstrate the use of micro-PIXE (proton induced X-ray emission) for measuring the quantitative uptake of anti-AIDS drugs, containing metal atoms, by individual Vero cells (African green monkey kidney cell line). Hetero-polytungstates, which are assessed to present an activity against the HIV virus, were studied using Vero cells. It was found that unlike other techniques, SPM offers both the sensitivity and the spatial resolution to carry out these programs of investigations. The use of elemental analysis in single cells of cultured cell lines has shown to have distinct advantages over peripheral blood lymphocytes. PMID:8833668

Cholewa, M; Turnbull, I F; Legge, G J; Weigold, H; Marcuccio, S M; Holan, G; Tomlinson, E; Wright, P J

1996-02-01

322

Compartmentation of metals in foliage of Populus tremula grown on soils with mixed contamination. I. From the tree crown to leaf cell level  

Microsoft Academic Search

In order to achieve efficient phytoextraction of heavy metals using trees, the metal allocation to aboveground tissues needs to be characterised. In his study, the distribution of heavy metals, macro- and micronutrients and the metal micro-localisation as a function of the leaf position and heavy metal treatment were analysed in poplars grown on soil with mixed metal contamination. Zinc was

Pierre Vollenweider; Terry Menard; Madeleine S. Günthardt-Goerg

2011-01-01

323

Compartmentation of metals in foliage of Populus tremula grown on soils with mixed contamination. II. Zinc binding inside leaf cell organelles  

Microsoft Academic Search

The phytoextraction potential of plants for removing heavy metals from polluted soils is determined by their capacity to store contaminants in aboveground organs and complex them safely. In this study, the metal compartmentation, elemental composition of zinc deposits and zinc complexation within leaves from poplars grown on soil with mixed metal contamination was analysed combining several histochemical and microanalytical approaches.

Pierre Vollenweider; Petra Bernasconi; Hans-Peter Gautschi; Terry Menard; Beat Frey; Madeleine S. Günthardt-Goerg

2011-01-01

324

Estrogen induction of progestophilins in rat estrogen-sensitive cells grown in media supplemented with sera from castrated rats and from rats bearing an alpha-fetoprotein-secreting hepatoma.  

PubMed

The purpose of this work was to study the effect of alpha-fetoprotein (AFP) over cell multiplication and the induction of an estradiol-17 beta (E2)-dependent marker, i.e., progestophilins in E-sensitive cells C2(9)RAP derived from a W/Fu rat pituitary tumor. These cells proliferate in isogeneic hosts under the influence of E2, while they proliferate in culture regardless of the presence of E2. C2(9)RAP cells were grown in medium supplemented with 10% horse serum. Progestophilin levels were measured 48 h after adding serum (20% horse, or castrated rat, or AFP-secreting tumor-bearing rat) and estrogen to the 10% horse serum-supplemented medium in which the cells were growing. Maximal induction of progestophilins was obtained at 3 X 10(-10) M E2 in cells grown in medium containing horse or castrated rat serum. In contrast, maximal induction of progestophilins required 3 X 10(-8) M E2 in cells grown in medium supplemented with the serum of Morris hepatoma 7777-bearing rats. This serum contained AFP levels comparable to those present at birth in the rat. 11-Methoxy-17 beta ethynylestradiol (R2858), a synthetic estrogen with little affinity for AFP, was also tested for its ability to induce progestophilins. The degree of maximal induction of progestophilins expressed as percentage of the respective control, was similar for all experimental groups, both with E2 and with R2858. In addition, we compared the free E2 levels in the culture medium with the progestophilin levels and the cell proliferation rate. We found that the progestophilin levels were maximal at free E2 concentrations above 11 pg E2/ml, whereas there was no correlation between the free E2 levels and the proliferation rate. Moreover, the proliferation rate of cells in medium supplemented with horse or castrated rat serum was maximal at concentrations of free E2 below 0.4 pg/ml; whereas cell proliferation was inhibited with hepatoma serum even at concentrations of free E2 of 44 pg/ml. We conclude that the effect of hepatoma serum on the E2 induction of progestophilins seems to be mediated by the effect of AFP on the availability of free estrogen, since it is abolished by the addition of both natural and synthetic estrogens. The inhibitory effect of hepatoma serum upon cell proliferation is not reversed by estrogens and thus seems to be mediated by mechanisms other than E2 trapping by AFP. PMID:6198194

Soto, A M; Lee, H; Siiteri, P K; Murai, J T; Sonnenschein, C

1984-02-01

325

Isolation and identification of a novel chlorophenol from a cell suspension culture of Helichrysum aureonitens.  

PubMed

A novel chlorophenol, 4-chloro-2-(hepta-1,3,5-triyn-1-yl)-phenol (1), was isolated as the major phenolic compound from the cells of Helichrysum aureonitens suspension cultures. Compound 1 has been proposed to be an intermediate in the acetylene biosynthetic pathway of other acetylenic compounds in Helichrysum spp. The ethanol extract of cell suspension cultures and compound 1 were evaluated for their cytotoxicity against monkey kidney Vero (Vero cells) and human prostate epithelial carcinoma (DU145) cell lines, also, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against Mycobacterium tuberculosis H37Rv were determined as well. PMID:19881282

Ziaratnia, Seyed Mahdi; Ohyama, Kiyoshi; Hussein, Ahmed Abdel-Fattah; Muranaka, Toshiya; Lall, Namrita; Kunert, Karl Josef; Meyer, Jacobus Johannes Marion

2009-11-01

326

Anchorage-dependent mammalian cell culture using polyurethane foam as a new substratum for cell attachment  

Microsoft Academic Search

Anchorage-dependent mammalian cells were cultivated at high cell density in a novel culture system using polyurethane foam (PUF) as a substratum for cell attachment. PUF has a macroporous structure giving a high surface area to volume ratio. Monkey kidney cells (Vero) and Chinese hamster ovary cells (CHO-K1) attached to the internal surface of PUF and grew to a high cell

Taku Matsushita; Masayoshi Ketayama; Ken-ichi Kamihata; Kazumori Funatsu

1990-01-01

327

Effect of black tea extract on herpes simplex virus-1 infection of cultured cells  

PubMed Central

Background The purpose of this investigation was to determine if black tea extract (BTE), consisting primarily of flavanol compounds called theaflavins, could inhibit herpes simplex virus type-1 (HSV-1) infection in cultured A549 (human epithelial) and Vero cells. Methods The effect of BTE both on A549 and Vero cultured cells and on HSV-1 was assessed by using phase contrast and fluorescent microscopy, and cell viability and proliferation assays. After establishing the maximum non-cytotoxic concentration of BTE, A549 and Vero cells and HSV-1 virions were treated with varying concentrations of BTE, respectively. A549 and Vero cells were infected with HSV-1 with green fluorescent protein (GFP) insert at the UL46 gene. The effect of infectivity was determined by viral DNA extraction followed by PCR, plaque assays, adsorption assays, and electrophoresis of PCR products. Results BTE was not cytotoxic to A549 and Vero cells, as confirmed by cell viability and proliferation assays, in which BTE treated groups paralleled the positive control group. For both cell lines, plaque assays and fluorescent microscopy indicated an inverse relationship between BTE concentration (from 0.14 ?M – 1.4 mM) and HSV-1 infectivity. Specifically, PCR and electrophoresis showed a reduction in the viral genome following treatment with BTE. In addition, there was a noticeable decrease in the amount of viral plaques for BTE treated samples in the adsorption assays. Conclusions BTE consisting primarily of theaflavins is not cytotoxic and can reduce or block the production of infectious HSV-1 virions in cultured A549 and Vero cells, thus inhibiting the infectivity of the virus by interfering in the attachment, penetration and viral DNA replication of HSV-1 particles. These findings indicate that BTE enriched with theaflavins has the potential to be developed as a safe, therapeutic antiviral agent to prevent the spread of HSV-1.

2013-01-01

328

Establishment of cell lines with increased susceptibility to EV71/CA16 by stable overexpression of SCARB2  

PubMed Central

Background Human enterovirus type 71 (EV71) and Coxsackievirus A group type 16 (CA16) belong to human Enterovirus species A of the family Picornaviridae. These viruses are recognized as the major pathogens responsible for epidemics of hand-foot-mouth disease (HFMD), which presents with fever and vesicular eruptions of palms, soles of the feet or mouth. Human scavenger receptor class B, member 2 (SCARB2) has been identified as the receptor for both EV71 and CA16, as overexpression of SCARB2 in cells can enhance virus replication significantly. Methods In this study, we used a lentivirus packaging vector to transduce the SCARB2 gene into human embryonic kidney cells (293), human rhabdomyosarcoma cells (RD) and African green monkey kidney cells (Vero) to create stable expression lines. Expression of SCARB2 in the resulting three transgenic cell lines was confirmed by real-time RT-PCR, immunofluorescence and flow cytometry. Results Levels of SCARB2 mRNA determined by real-time RT-PCR in 293-SCARB2 (293S) or RD-SCARB2 (RDS) transgenic cell lines were approximately 2?×?102 times higher than those in 293 and RD cells, respectively, and three times higher in Vero-SCARB2 (VeroS) than in Vero cells. Furthermore, EV71 and CA16 virus titers in 293S and RDS cells were 102–103-fold higher (detected in RD cell) than those in the parental cells, and a 10-fold higher titer of EV71 was achieved in VeroS cells compared with that in Vero cells. Conclusions We established for the first time three cell lines stably overexpressing SCARB2, which showed drastic increases in susceptibility to EV71/CA16 infection. These optimal cell lines may be utilized to develop inactivated vaccines for EV71/CA16 and facilitate rapid detection and isolation of HFMD pathogens or other Enterovirus serotypes. Furthermore, these stable cell lines also can serve as tools to facilitate drug screenings as well as molecular studies on virus-host interactions and pathogenesis of causative agents for HFMD.

2013-01-01

329

Bi2S3microspheres grown on graphene sheets as low-cost counter-electrode materials for dye-sensitized solar cells  

NASA Astrophysics Data System (ADS)

In this work, we synthesized 3D Bi2S3 microspheres comprised of nanorods grown along the (211) facet on graphene sheets by a solvothermal route, and investigated its catalytic activities through I-V curves and conversion efficiency tests as the CE in DSSCs. Although the (211) facet has a large band gap for a Bi2S3 semiconductor, owing to the introduction of graphene into the system, its short-circuit current density, open-circuit voltage, fill factor, and efficiency were Jsc = 12.2 mA cm-2, Voc = 0.75 V, FF = 0.60, and ? = 5.5%, respectively. By integrating it with graphene sheets, our material achieved the conversion efficiency of 5.5%, which is almost triple the best conversion efficiency value of the DSSCs with (211)-faceted 3D Bi2S3 without graphene (1.9%) reported in the latest literature. Since this conversion-efficient 3D material grown on the graphene sheets significantly improves its catalytic properties, it paves the way for designing and applying low-cost Pt-free CE materials in DSSC from inorganic nanostructures.In this work, we synthesized 3D Bi2S3 microspheres comprised of nanorods grown along the (211) facet on graphene sheets by a solvothermal route, and investigated its catalytic activities through I-V curves and conversion efficiency tests as the CE in DSSCs. Although the (211) facet has a large band gap for a Bi2S3 semiconductor, owing to the introduction of graphene into the system, its short-circuit current density, open-circuit voltage, fill factor, and efficiency were Jsc = 12.2 mA cm-2, Voc = 0.75 V, FF = 0.60, and ? = 5.5%, respectively. By integrating it with graphene sheets, our material achieved the conversion efficiency of 5.5%, which is almost triple the best conversion efficiency value of the DSSCs with (211)-faceted 3D Bi2S3 without graphene (1.9%) reported in the latest literature. Since this conversion-efficient 3D material grown on the graphene sheets significantly improves its catalytic properties, it paves the way for designing and applying low-cost Pt-free CE materials in DSSC from inorganic nanostructures. Electronic supplementary information (ESI) available. See DOI: 10.1039/c3nr06093d

Li, Guang; Chen, Xiaoshuang; Gao, Guandao

2014-02-01

330

Deep-level transient spectroscopy characterization of In 0.48Ga 0.52P grown at different V/III ratio using a valved phosphorus cracker cell in solid source molecular beam epitaxy  

NASA Astrophysics Data System (ADS)

In 0.48Ga 0.52P/n +GaAs structures have been grown by solid source molecular beam epitaxy (SSMBE) on GaAs(1 0 0) substrates using a valved phosphorus and arsenic cracker cell. The In 0.48Ga 0.52P layer grown using this technique was characterized using deep-level transient spectroscopy (DLTS) measurements to investigate the correlation between the V/III ratio used in In 0.48Ga 0.52P growth and the defect characteristics. Under typical growth conditions used for the In 0.48Ga 0.52P/n +GaAs/GaAs structures, the trap concentration increased from 0.16±0.008×10 14 to 1.48±0.074×10 14 cm -3 when the V/III flux ratio was increased from 10 to 50. The activation energy for the electron trap decreased from ˜0.436±0.004 to 0.307±0.008 eV, and saturation above V/III ratio of 40 occurred. There was a corresponding decrease in the electron capture cross section from 6.3×10 -14 to 0.015×10 -14 cm 2. The results of this study have important implications for the growth of In 0.48Ga 0.52P by SSMBE using the valved phosphorus cracker cell technique.

Yoon, S. F.; Lui, P. Y.; Zheng, H. Q.

1999-05-01

331

31P magnetic resonance spectroscopy of endothelial cells grown in three-dimensional matrigel constructs as an enabling platform technology: II. The effect of anti-inflammatory drugs on phosphometabolite levels.  

PubMed

In the accompanying study, the authors presented phosphometabolite patterns of endothelial cells grown under three-dimensional (3D) conditions using (31)P magnetic resonance spectroscopy (MRS). Here the authors describe the effect of nonsteroidal anti-inflammatory drugs (NSAIDs), using this enabling platform technology, which is relevant for evaluating drug effects in tissue-engineered endothelial constructs. Treatment with indomethacin significantly changed the phosphometabolite fingerprint in this endothelial model, by, respectively, increasing (81%) and decreasing (42%) glycerophosphocholine (GPC) and phosphomonoesters (PM). Furthermore, a safer approach using a NSAID prodrug was also demonstrated in this study with a indomethacin phospholipid-derived prodrug (DP-155). Like the parental drug, DP-155 increased and decreased the levels of GPC and PM by 100% and 20%, respectively. These changes represent useful biomarkers to monitor NSAID effects on endothelized tissue-engineered constructs for the purpose of controlling endothelial cell survival and inflammation upon implantation. PMID:19065321

Ringel, I; Lecht, S; Sterin, M; Lelkes, P I; Lazarovici, P

2008-01-01

332

Differential Effect of Auxin on Molecular Weight Distributions of Xyloglucans in Cell Walls of Outer and Inner Tissues from Segments of Dark Grown Squash (Cucurbita maxima Duch.) Hypocotyls 1  

PubMed Central

Effects of indole-3-acetic acid (IAA) on the mechanical properties of cell walls and structures of cell wall polysaccharides in outer and inner tissues of segments of dark grown squash (Cucurbita maxima Duch.) hypocotyls were investigated. IAA induced the elongation of unpeeled, intact segments, but had no effect on the elongation of peeled segments. IAA induced the cell wall loosening in outer tissues as studied by the stress-relaxation analysis but not in inner tissues. IAA-induced changes in the net sugar content of cell wall fractions in outer and inner tissues were very small. Extracted hemicellulosic xyloglucans derived from outer tissues had a molecular weight about two times as large as in inner tissues, and the molecular weight of xyloglucans in both outer and inner tissues decreased during incubation. IAA substantially accelerated the depolymerization of xyloglucans in outer tissues, while it prevented that in inner tissues. These results suggest that IAA-induced growth in intact segments is due to the cell wall loosening in outer tissues, and that IAA-accelerated depolymerization of hemicellulosic xyloglucans in outer tissues is involved in the cell wall loosening processes.

Wakabayashi, Kazuyuki; Sakurai, Naoki; Kuraishi, Susumu

1991-01-01

333

Free amino acid and phenolic contents and antioxidative and cancer cell-inhibiting activities of extracts of 11 greenhouse-grown tomato varieties and 13 tomato-based foods.  

PubMed

Tomato (Solanum lycopersicum) plants synthesize nutrients, pigments, and bioactive compounds that benefit nutrition and human health. The nature and concentrations of these compounds are strongly influenced by varietal factors such as size and color as well as by processing. To better understand how these factors affect the concentration of nutrients and bioactive compounds, we analyzed 11 Korean tomato varieties grown under the same greenhouse conditions and 13 processed commercial tomato products for free amino acids and amino acid metabolites by HPLC, for individual phenolics by HPLC-MS, for total phenolics by the Folin-Ciocalteu method, for antioxidative activity by the FRAP and DPPH methods, and for cancer cell-inhibiting effects by the MTT assay. We also determined the protein content of the tomatoes by an automated Kjeldahl method. The results show that there is a broad range of bioactive compounds across tomato varieties and products. Small tomatoes had higher contents of bioactive compounds than the large ones. The content of phenolic compounds of processed products was lower than that of fresh tomatoes. Tomato extracts promoted growth in normal liver (Chang) cells, had little effect in normal lung (Hel299) cells, mildly inhibited growth of lung cancer (A549) cells, and first promoted and then, at higher concentrations, inhibited growth in lymphoma (U937) cells. The relationship of cell growth to measured constituents was not apparent. Dietary and health aspects of the results are discussed. PMID:22070764

Choi, Suk-Hyun; Kim, Hyen-Ryung; Kim, Hyun-Jeong; Lee, In-Seon; Kozukue, Nobuyuki; Levin, Carol E; Friedman, Mendel

2011-12-28

334

Scanning electron microscopic study of human neuroblastoma cells affected with Naegleria fowleri Thai strains.  

PubMed

In order to understand the pathogenesis of Naegleria fowleri in primary amoebic meningoencephalitis, the human neuroblastoma (SK-N-MC) and African green monkey kidney (Vero) cells were studied in vitro. Amoeba suspension in cell-culture medium was added to the confluent monolayer of SK-N-MC and Vero cells. The cytopathic activity of N. fowleri trophozoites in co-culture system was elucidated by scanning electron microscope at 3, 6, 9, 12, and 24 h. Two strains of N. fowleri displayed well-organized vigorous pseudopods in Nelson's medium at 37 degrees C. In co-culture, the target monolayer cells were damaged by two mechanisms, phagocytosis by vigorous pseudopods and engulfment by sucker-like apparatus. N. fowleri trophozoites produced amoebostomes only in co-culture with SK-N-MC cells. In contrast, we could not find such apparatus in the co-culture with Vero cells. The complete destruction time (100%) at 1:1 amoeba/cells ratio of SK-N-MC cells (1 day) was shorter than the Vero cells (12 days). In conclusion, SK-N-MC cells were confirmed to be a target model for studying neuropathogenesis of primary amoebic meningoencephalitis. PMID:18685867

Tiewcharoen, Supathra; Rabablert, Jundee; Chetanachan, Pruksawan; Junnu, Virach; Worawirounwong, Dusit; Malainual, Nat

2008-10-01

335

Plaque Assay of Rickettsiae in a Mammalian Cell Line  

PubMed Central

Clear-cut and repeatable plaque assays were obtained for three rickettsiae of the spotted fever group (Rickettsia rickettsi, R. conori, and R. montana) in Vero cells used in a manner similar to that for arboviruses. In addition, three typhus group agents (R. typhi, R. canada, R. prowazeki) induced plaques in these cells. In preliminary tests Coxiella burneti (Nine Mile strain) failed to produce plaques. Comparable results were obtained in plastic flasks and plastic culture trays incubated in ambient air with or without addition of N-2-hydroxyethyl-piperazine-N?-2-ethanesulfinic acid buffer. Larger and more well defined R. rickettsi plaques were produced when cultures were overlaid with Leibovitz (L15) medium than with either medium 199 or Eagle medium. Phosphate-buffered saline containing bovine plasma albumin (fraction V), in contrast to brain heart infusion broth, as a diluent for preparing inocula consistently permitted development of larger and more numerous plaques with three agents: R. rickettsi, R. conori, and R. montana. When R. rickettsi and R. typhi were assayed in parallel in primary chicken embryo cultures and Vero cells, comparable results were obtained, but with R. canada results in Vero cells were superior. In contrast, R. prowazeki produced inconsistent results in Vero cells. Images

Cory, J.; Yunker, C. E.; Ormsbee, R. A.; Peacock, M.; Meibos, H.; Tallent, G.

1974-01-01

336

Plaque assay of rickettsiae in a mammalian cell line.  

PubMed

Clear-cut and repeatable plaque assays were obtained for three rickettsiae of the spotted fever group (Rickettsia rickettsi, R. conori, and R. montana) in Vero cells used in a manner similar to that for arboviruses. In addition, three typhus group agents (R. typhi, R. canada, R. prowazeki) induced plaques in these cells. In preliminary tests Coxiella burneti (Nine Mile strain) failed to produce plaques. Comparable results were obtained in plastic flasks and plastic culture trays incubated in ambient air with or without addition of N-2-hydroxyethyl-piperazine-N'-2-ethanesulfinic acid buffer. Larger and more well defined R. rickettsi plaques were produced when cultures were overlaid with Leibovitz (L15) medium than with either medium 199 or Eagle medium. Phosphate-buffered saline containing bovine plasma albumin (fraction V), in contrast to brain heart infusion broth, as a diluent for preparing inocula consistently permitted development of larger and more numerous plaques with three agents: R. rickettsi, R. conori, and R. montana. When R. rickettsi and R. typhi were assayed in parallel in primary chicken embryo cultures and Vero cells, comparable results were obtained, but with R. canada results in Vero cells were superior. In contrast, R. prowazeki produced inconsistent results in Vero cells. PMID:4208640

Cory, J; Yunker, C E; Ormsbee, R A; Peacock, M; Meibos, H; Tallent, G

1974-06-01

337

Characterization of peroxisomes in glucose-grown Hansenula polymorpha and their development after the transfer of cells into methanol-containing media  

Microsoft Academic Search

Cells of Hansenula polymorpha growing exponentially on glucose generally contained a single peroxisome of small dimension, irregular in shape and located in close proximity to the cell wall. Crystalline inclusions in the peroxisomal matrix were not observed. Associations of the organelles with one or more strands of endoplasmic reticulum were evident. In stationary phase cells the size of the peroxisomes

M. Veenhuis; I. Keizer; W. Harder

1979-01-01

338

Effects of Sodium Fluoride on Uptake of T-2 Mycotoxin in Cultured Cells,  

National Technical Information Service (NTIS)

The authors examined the effect of sodium fluoride on uptake of tritium-labeled T-2 toxin (molecules of toxin/cell) in Chinese hamster ovary (CHO) and African green monkey kidney (VERO) cells. Correlations were made to temperature (22 and 37 C) and toxin ...

L. R. Trusal L. J. Martin

1987-01-01

339

Lipid peroxidation and activities of tyrosine aminotransferase and glutamine synthetase in hepatoma and glioma cells grown in bovine colostrum-supplemented medium  

Microsoft Academic Search

Summary  The growth stimulating properties of bovine serum and colostrum were compared in rat hepatoma (HTC) and glioma (C6) cell cultures.\\u000a A colostrum concentration of 2% was optimal for HTc cells, which then reached a terminal density 40% of that in serum-supplemented\\u000a medium. The corresponding figures for C6 cells were 10 and 81%, respectively. After 4 d in culture, levels of

Lena Odland; Stefan Wallin; Erik Walum

1986-01-01

340

Comparative lipid composition of heterotrophically and autotrophically grown Sulfolobus acidocaldarius.  

PubMed Central

Complex lipids from the thermoacidophilic facultative autotroph Sulfolobus acidocaldarius, as well as a strictly autotrophic isolate, were compared between cells grown on yeast extract and elemental sulfur. Lipids from both organisms grown autotrophically were nearly identical. Each contained about 15% neutral lipids, 35% glycolipids, and 50% acidic lipids. Glycolipids and acidic lipids contained C40H82-76-derived glycerol ether residues. Major glycolipids included the glycerol ether analogues of glucosyl galactosyl diglyceride (5%) and glucosyl polyol diglyceride (75%). Acidic lipids were comprised mainly of the glycerol ether analogues of phosphatidyl inositol (7%), inositolphosphoryl glucosyl polyol diglyceride (72%), and a partially characterized sulfate- and phosphate-containing derivative of glucosyl polyol diglyceride (13%). The lipids from cells grown heterotrophically were similar to those from autotrophically grown cells, except that the partially characterized acidic lipid was absent. In addition, the two glycolipids as well as the respective inositolphosphoryl derivatives were each present in nearly equal proportions. Images

Langworthy, T A

1977-01-01

341

Osmotic Properties of Spheroplasts from Saccharomyces cerevisiae Grown at Different Temperatures  

PubMed Central

Spheroplasts were prepared from cells of Saccharomyces cerevisiae NCYC 366, grown at 30 or 15 C, by incubating cells with snail-gut juice after pretreatment with 2-mercaptoethanol. Walls of cells grown batchwise or in continuous culture at 15 C were more resistant to digestion with snail juice than walls on cells grown under the same conditions as 30 C. Spheroplasts lysed when suspended in hypotonic solutions of mannitol. The resistance of spheroplasts to osmotic lysis tended to increase when the test temperature was lowered below 30 C. The increased resistance was greater with spheroplasts from cells grown at 15 C. Cations, especially Ca2+, protected spheroplasts against osmotic lysis. In general, the protective effects, measured at 30 C, were smaller with spheroplasts from cells grown at 15 C compared with 30 C. Citrate and ethylenediaminetetraacetate (EDTA) decreased the resistance of spheroplasts to osmotic lysis. On the whole, the decrease was greater with spheroplasts from cells grown at 30 C rather than 15 C. In the presence of EDTA, spheroplasts from cells grown at 30 C were less resistant to osmotic lysis at 5 C than at 30 C; when spheroplasts from cells grown at 15 C were similarly examined, they were more resistant to lysis at 5 C than at 30 C. Spheroplast membranes from cells grown at 15 C had slightly but significantly greater contents of Mg2+, Ca2+, K+, and Na+ compared with spheroplast membranes from cells grown at 15 C. Mg2+ and Ca2+ were more easily extracted with EDTA from membranes of 30 C-grown cells than from 15 C-grown cells.

Diamond, R. J.; Rose, A. H.

1970-01-01

342

Proteolytic enzymes in embryonated chicken eggs sustain the replication of egg-grown low-pathogenicity avian influenza viruses in cells in the absence of exogenous proteases.  

PubMed

Low pathogenic influenza viruses grow readily in embryonated chicken eggs but require the addition of exogenous proteases to grow in MDCK cell culture. In this study, we found that the influenza viruses propagated previously in eggs, can grow for up to two passages in cell culture without the addition of exogenous proteolytic enzymes. These results indicate that the reason for virus propagation in cells during the first two passages may be due to proteases from egg allantoic fluid carried over from egg culture. The ability of influenza viruses to grow in cells in the absence of trypsin is currently considered as a hallmark of highly pathogenic influenza viruses. Our data indicate that differentiating between high and low pathogenicity using cell culture only is not appropriate and other indicators such as sequence analysis and in vitro pathogenicity index should be performed. PMID:24626064

Kandeil, Ahmed; Bagato, Ola; Zaraket, Hassan; Debeauchamp, Jennifer; Krauss, Scott; El-Shesheny, Rabeh; Webby, Richard J; Ali, Mohamed A; Kayali, Ghazi

2014-06-15

343

RICKETTSIAL PHOSPHOLIPASE A 2 AS A PATHOGENIC MECHANISM IN A MODEL OF CELL INJURY BY TYPHUS AND SPOTTED FEVER GROUP RICKETTSIAE  

Microsoft Academic Search

Phospholipase A2 activity by typhus group rickettsiae causes hemolysis in vitro. Rickettsial phospholi- pase A2 has been proposed to mediate entry into the host cell, escape from the phagosome, and cause injury to host cells by both typhus and spotted fever group rickettsiae. In a rickettsial contact-associated cytotoxicity model, the interaction of Rickettsia prowazekii or R. conorii with Vero cells

DAVID H. WALKER; HUI-MIN FENG; VSEVOLOD L. POPOV

2001-01-01

344

Standardization and assessment of cell culture media quantities in roller poly ethylene terephthalate bottles employed in the industrial rabies viral vaccine production.  

PubMed

Vero cells are utilized for production of rabies vaccine. This study deals with the optimize quantity media require for the rabies vaccine production in the smooth roller surface. The rabies virus (Pasteur vaccine strain) is infected to monolayer of the various experimented bottles. To analyze the optimal quantity of media for the production of rabies viral harvest during the process of Vero cell derived rabies vaccine. The trials are started from 200 to 400 mL (PTARV-1, PTARV-2, PTARV-3, PTARV-4 and PTARV-5). The samples are taken in an appropriate time intervals for analysis of In Process Quality Control (IPQC) tests. The collected viral harvests are further processed to rabies vaccine in a pilot level and in addition to scale up an industrial level. Based on the evaluation the PTARV-2 (250 mL) show highly encouraging results for the Vero cell derived rabies vaccine production. PMID:20384277

Jagannathan, S; Chaansha, S; Rajesh, K; Santhiya, T; Charles, C; Venkataramana, K N

2009-09-15

345

Factors Influencing Synthesis and Mineralization of Bone Matrix from Fetal Bovine Bone Cells Grown In vitro. (Reannouncement with New Availability Information).  

National Technical Information Service (NTIS)

This study of the in vitro synthesis and mineralization of bovine bone demonstrates that sheets of mineralized matrix can be produced consistently within 18-24 days of cell isolation. Mineralization surpasses that achieved by other systems with other spec...

S. W. Whitson M. A. Whitson D. E. Bowers M. C. Falk

1992-01-01

346

N/P InP homojunction solar cells with an In0.53Ga0.47As contacting layer grown by liquid phase epitaxy  

NASA Technical Reports Server (NTRS)

N/P InP homojunction solar cells with an In sub 0.53 Ga sub 0.47 As contacting layer were fabricated by liquid phase epitaxy (LPE). Electron-Beam-Induced-Current (EBIC) measurements were performed on several selected samples. It was found that the background doping level in the base region sometimes results in a deep junction, which greatly affects the cell performance.

Shen, C. C.; Choi, K. Y.

1989-01-01

347

Morphological and molecular changes in the unicellular green alga Dunaliella salina grown under supplemental UV-B radiation: cell characteristics and Photosystem II damage and repair properties  

Microsoft Academic Search

The effect of supplemental UV-B radiation during growth in the green alga Dunaliella salina was investigated. At the cellular level, supplemental UV-B radiation induced a doubling of the cell volume, a phenomenon attributed to a slow-down in the rate of cell division. At the thylakoid mebrane level, supplemetal UV-B radiation induced photodamage to the 32 kDa (D1) and 34 kDa

Antonio Masi; Anastasios Melis

1997-01-01

348

Human Umbilical Cord Wharton’s Jelly Stem Cells Undergo Enhanced Chondrogenic Differentiation when Grown on Nanofibrous Scaffolds and in a Sequential Two-stage Culture Medium Environment  

Microsoft Academic Search

The current treatments used for osteoarthritis from cartilage damage have their disadvantages of donor site morbidity, complicated\\u000a surgical interventions and risks of infection and graft rejection. Recent advances in tissue engineering have offered much\\u000a promise in cartilage repair but the best cell source and in vitro system have not as yet been optimised. Human bone marrow\\u000a mesenchymal stem cells (hBMSCs)

Chui-Yee Fong; Arjunan Subramanian; Kalamegam Gauthaman; Jayarama Venugopal; Arijit Biswas; Seeram Ramakrishna; Ariff Bongso

349

Feasibility evaluation of a new irradiation technique: three-dimensional unicursal irradiation with the Vero4DRT (MHI-TM2000)  

PubMed Central

The Vero4DRT (MHI-TM2000) is a newly designed unique image-guided radiotherapy system consisting of an O-ring gantry. This system can realize a new irradiation technique in which both the gantry head and O-ring continuously and simultaneously rotate around the inner circumference of the O-ring and the vertical axis of the O-ring, respectively, during irradiation. This technique creates three-dimensional (3D) rotational dynamic conformal arc irradiation, which we term ‘3D unicursal irradiation’. The aim of this study was to present the concept and to estimate feasibility and potential advantages of the new irradiation technique. Collision maps were developed for the technique and a 3D unicursal plan was experimentally created in reference to the collision map for a pancreatic cancer case. Thereafter, dosimetric comparisons among the 3D unicursal, a two-dimensionally rotational dynamic conformal arc irradiation (2D–DCART), and an intensity-modulated radiation therapy (IMRT) plan were conducted. Dose volume data of the 3D unicursal plan were comparable or improved compared to those of the 2D–DCART and IMRT plans with respect to both the target and the organs at risk. The expected monitor unit (MU) number for the 3D unicursal plan was only 7% higher and 22.1% lower than the MUs for the 2D–DCART plan and IMRT plan, respectively. It is expected that the 3D unicursal irradiation technique has potential advantages in both treatment time and dose distribution, which should be validated under various conditions with a future version of the Vero4DRT fully implemented the function.

Mizowaki, Takashi; Takayama, Kenji; Nagano, Kazuo; Miyabe, Yuki; Matsuo, Yukinori; Kaneko, Shuji; Kokubo, Masaki; Hiraoka, Masahiro

2013-01-01

350

Prostate tumor grown in NASA Bioreactor  

NASA Technical Reports Server (NTRS)

This prostate cancer construct was grown during NASA-sponsored bioreactor studies on Earth. Cells are attached to a biodegradable plastic lattice that gives them a head start in growth. Prostate tumor cells are to be grown in a NASA-sponsored Bioreactor experiment aboard the STS-107 Research-1 mission in 2002. Dr. Leland Chung of the University of Virginia is the principal investigator. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. Credit: NASA and the University of Virginia.

2001-01-01

351

Antibacterial effect of theaflavin, polyphenon 60 ( Camellia sinensis) and Euphorbia hirta on Shigella spp. — a cell culture study  

Microsoft Academic Search

Antibacterial effect of compounds extracted from Camellia sinensis L. and the methanol extract of Euphorbia hirta L. were studied against dysentery causing Shigella spp. using the Vero cell line. Cytotoxicity studies of the extracts were performed using the cell line and the non-cytotoxic concentration of the extract was tested for antibacterial activity against the cytopathic dose of the pathogen. These

K. Vijaya; S. Ananthan; R. Nalini

1995-01-01

352

Protein and phosphoprotein levels in glioma and adenocarcinoma cell lines grown in normoxia and hypoxia in monolayer and three-dimensional cultures  

PubMed Central

Background Three dimensional (3D) growths of cancer cells in vitro are more reflective of in situ cancer cell growth than growth in monolayer (2D). The present study is designed to determine changes in protein and phosphoprotein that reflect adaptation of tumor cells to 3D as compared to 2D. Since relative hypoxia is a common feature of most solid tumors, the present study also aims to look at the impact of transition from normoxia to hypoxia in these two growth conditions. Results Using reverse-phase protein arrays, we compared levels of 121 different phosphorylated and non-phosphorylated proteins in 5 glioma and 6 adenocarcinoma lines under conditions of 3D and monolayer culture in normoxia and hypoxia. A three-way analysis of variance showed levels of 82 antibodies differed between media (2D vs. 3D) and 49 differed between treatments (hypoxia vs. normoxia). Comparing 2D to 3D growth, 7 proteins were commonly (i.e., > 50% of tumors) elevated in 3D: FAK, AKT, Src, GSK3??, TSC2, p38, and NF??p65. Conversely, 7 other proteins are commonly decreased: ATRIP, ATR, ?-catenin, BCL-X, cyclin B1, Egr-1, and HIF-1?. Comparing normoxia to hypoxia, only NCKIPSD was commonly elevated in hypoxia; 6 proteins were decreased: cyclin B1, 4EBP1(Ser65), c-Myc, SMAD3(Ser423), S6(Ser235), and S6(Ser240). Hypoxia affected glioma cell lines differently from adenocarcinoma cell lines: 8 proteins were increased in gliomas (BAX, caspase 7, HIF-1?, c-JUN, MEK1, PARP 1 cleaved, Src, and VEGFR2) and none in adenocarcinomas. Conclusions We identified subsets of proteins with clearly concordant/discordant behavior between gliomas and adenocarcinomas. In general, monolayer to 3D culture differences are clearer than normoxia to hypoxia differences, with anti-apoptotic, cytoskeletal rearrangement and cell survival pathways emphasized in the former and mTOR pathway, transcription, cell-cycle arrest modulation, and increased cell motility in the latter.

2012-01-01

353

Epitaxial lead titanate grown by MBE  

NASA Astrophysics Data System (ADS)

Epitaxial PbTiO 3 films have been grown by reactive molecular beam epitaxy (MBE) on (100) LaAlO 3 substrates. Lead is supplied from a conventional effusion cell. Titanium is sublimated from a Ti-Ball™, and oxygen is supplied in the form of purified ozone. Atomic layer-by-layer composition control is obtained using real-time atomic absorption spectroscopy (AA) feedback. The titanium flux is controlled with a shutter directly coupled to the titanium AA feedback to deliver a burst corresponding to one monolayer of titania. Similarly, lead is monitored in situ using an atomic absorption signal. An adsorption-controlled growth mechanism leads to the use of a lead overpressure to insure films with proper stoichiometries. Film structure is studied during growth using in situ RHEED. 4-circle X-ray diffraction analysis indicates that films grown on LaAlO 3 are epitaxial and are mixed a- and c-axis oriented.

Theis, C. D.; Schlom, D. G.

1997-04-01

354

Quantification of confocal images of biofilms grown on irregular surfaces.  

PubMed

Bacterial biofilms grow on many types of surfaces, including flat surfaces such as glass and metal and irregular surfaces such as rocks, biological tissues and polymers. While laser scanning confocal microscopy can provide high-resolution images of biofilms grown on any surface, quantification of biofilm-associated bacteria is currently limited to bacteria grown on flat surfaces. This can limit researchers studying irregular surfaces to qualitative analysis or quantification of only the total bacteria in an image. In this work, we introduce a new algorithm called modified connected volume filtration (MCVF) to quantify bacteria grown on top of an irregular surface that is fluorescently labeled or reflective. Using the MCVF algorithm, two new quantification parameters are introduced. The modified substratum coverage parameter enables quantification of the connected-biofilm bacteria on top of the surface and on the imaging substratum. The utility of MCVF and the modified substratum coverage parameter were shown with Pseudomonas aeruginosa and Staphylococcus aureus biofilms grown on human airway epithelial cells. A second parameter, the percent association, provides quantified data on the colocalization of the bacteria with a labeled component, including bacteria within a labeled tissue. The utility of quantifying the bacteria associated with the cell cytoplasm was demonstrated with Neisseria gonorrhoeae biofilms grown on cervical epithelial cells. This algorithm provides more flexibility and quantitative ability to researchers studying biofilms grown on a variety of irregular substrata. PMID:24632515

Sommerfeld Ross, Stacy; Tu, Mai Han; Falsetta, Megan L; Ketterer, Margaret R; Kiedrowski, Megan R; Horswill, Alexander R; Apicella, Michael A; Reinhardt, Joseph M; Fiegel, Jennifer

2014-05-01

355

Photocurrent enhancement in In0.53Ga0.47As solar cells grown on InP\\/SiO2\\/Si transferred epitaxial templates  

Microsoft Academic Search

InP\\/Si engineered substrates formed by wafer bonding and layer transfer have the potential to significantly reduce the cost and weight of III-V compound semiconductor solar cells. InP\\/Si substrates were prepared by He implantation of InP prior to bonding to a thermally oxidized Si substrate and annealing to exfoliate an InP thin film. Following thinning of the transferred InP film to

James M. Zahler; Katsuaki Tanabe; Corinne Ladous; Tom Pinnington; Frederick D. Newman; Harry A. Atwater

2007-01-01

356

Improvement in photovoltaic conversion efficiency of InGaP solar cells grown on Si substrate by thermal cleaning using Si 2H 6  

Microsoft Academic Search

A new type of thermal cleaning for Si surfaces, using Si2H6, has been developed for growing GaAs buffer layers with an A-step surface on an Si substrate by metaloraganic vapor-phase epitaxy (MOVPE). This process made it possible, for the first time, to grow an A-step surface InGaP solar cell structure on an Si substrate with good surface morphology. An improvement

Shu Goto; Takashi Ueda; Chouho Yamagishi

2001-01-01

357

Dependence of the recombination in thin-film Si solar cells grown by ion-assisted deposition on the crystallographic orientation of the substrate  

Microsoft Academic Search

One promising strategy for achieving high-quality polycrystalline silicon thin-film solar cells on glass is based on low-temperature ion-assisted deposition for epitaxial thickening of a thin, large-grained seeding layer on glass. The crystal growth on the seeding layer is influenced by various factors, amongst which the crystal orientation of the grains plays a substantial role. In this paper we investigate how

D. H Neuhaus; N.-P Harder; S Oelting; R Bardos; A. B Sproul; P Widenborg; A. G Aberle

2002-01-01

358

Alterations of leaf cell ultrastructures and AFLP DNA profiles in Earth-grown tomato plants propagated from long-term six years Mir-flown seeds  

Microsoft Academic Search

Leaf cell ultrastructures and DNA variations in the firstand the second-generation of Earthgrown tomato (Lycopersicon esculentun Mill) plants that had been endured a long-term six years spaceflight in the Mir were compared to their ground-based control plants, under observations with a Transmission Electron Microscope and the Amplification Fragment Length Polymorphism (AFLP) analysis. For alterations in the morphological ultrastructures, one plant

Min Liu; Huai Xue; Yi Pan; Chunhua Zhang; Jinying Lu

2008-01-01

359

Vaccinia virus exhibits cell-type-dependent entry characteristics  

PubMed Central

Differing and sometimes conflicting data have been reported regarding several aspects of vaccinia virus (VV) entry. To address this, we used a ?-galactosidase reporter virus to monitor virus entry into multiple cell types under varying conditions. Entry into HeLa, B78H1 and L cells was strongly inhibited by heparin whereas entry into Vero and BSC-1 cells was unaffected. Bafilomycin also exhibited variable and cell-type-specific effects on VV entry. Entry into B78H1 and BSC-1 cells was strongly inhibited by bafilomycin whereas entry into Vero and HeLa cells was only partially inhibited suggesting the co-existence of both pH-dependent and pH-independent VV entry pathways in these cell types. Finally, entry into HeLa, B78H1, L and BSC-1 cells exhibited a lag of 6–9 min whereas this delay was undetectable in Vero cells. Our results suggest that VV exploits multiple cell attachment and entry pathways allowing it to infect a broad range of cells.

Whitbeck, J. Charles; Foo, Chwan-Hong; de Leon, Manuel Ponce; Eisenberg, Roselyn J.; Cohen, Gary H.

2014-01-01

360

Alterations of leaf cell ultrastructures and AFLP DNA profiles in Earth-grown tomato plants propagated from long-term six years Mir-flown seeds  

NASA Astrophysics Data System (ADS)

Leaf cell ultrastructures and DNA variations in the firstand the second-generation of Earthgrown tomato (Lycopersicon esculentun Mill) plants that had been endured a long-term six years spaceflight in the Mir were compared to their ground-based control plants, under observations with a Transmission Electron Microscope and the Amplification Fragment Length Polymorphism (AFLP) analysis. For alterations in the morphological ultrastructures, one plant among the 11 first-generation plants generated from 30 Mir-flown seeds had a three-layered palisade cell structure, while other 10 first-generation plants and all ground-based controls had one-layered palisade cell structure in leaves. Starch grains were larger and in clusters, numbers of starch grains increased in the chloroplasts in the Mir-flown plants. Leaf cells became contracted and deformed, and cell shape patterns were different in the Mir-flown plants. For the leaf genomic DNA alterations, 34 DNA bands were polymorphic with a 1.32% polymorphism among 2582 DNA bands in the first-generation Mir-flown plants. Band types in the spaceflight treated plants were also different from those in the ground-based control. Of 11 survived first-generation plants, 7 spaceflight treated plants (Plant Nos. 1-6 and No. 9) had a same 7 polymorphic bands and a same 0.27%DNA mutation. The DNA mutation rate was greatest in Plants No.10 and No.7 (0.90% and 0.94%), less in Plant No.11 (0.31%) and least in Plant No.8 (0.20%). For the 38 send-generation plants propagated from the No. 5 Mir-flown seed, 6 DNA bands were polymorphic with a 0.23% polymorphism among 2564 amplified DNA bands. Among those 38 second-generation plants amplified by primer pair (E4: ACC, M8: CTT), one DNA band disappeared in 29 second-generation plants and in the original Mir-flown No. 5 plant, compared to the ground-base controls. Among the 38 second-generation plants generated from the Mir-flown No. 5 seed, the DNA band types of 29 second-generation plants were different from that of the ground-base controls and had a same 6 polymorphic bands and a same 0.23% DNA mutation. For the 49 second-generation plants derived from the Mir-flown No. 6 seed, 7 DNA bands were polymorphic with 0.27% polymorphism among 2564 amplified DNA bands. With only one exception among those 49 second-generation plants amplified by primer pair (E3: ACA, M3: CAG), one DNA band disappeared in 48 second-generation plants and in the original Mir-flown No. 6 plant, compared to the ground-based controls. Among the 49 second-generation plants generated from the Mir-flown No. 6 seed, the DNA band types of 48 second-generation plants were different from that of the ground-base controls and had a same 7 polymorphic bands and a same 0.27% DNA mutation. Our results indicated that leaf cell ultrastructures had been altered and heredity variations had been induced by seeds being exposed to a long-term outer-space environment. Further research is needed to elucidate the dynamics and mechanisms resulting in such variations. Plant biology studies in the space environment may open potential approaches to induce mutations and to screen new plant varieties by ground-based selections among spaceflight treated seeds or seedlings.

Liu, Min; Xue, Huai; Pan, Yi; Zhang, Chunhua; Lu, Jinying

361

In situ grown vertically oriented CuInS2 nanosheets and their high catalytic activity as counter electrodes in dye-sensitized solar cells.  

PubMed

Vertically oriented CuInS(2) nanosheet thin films were prepared via a facile one-step solvothermal process and used in dye-sensitized solar cells (DSSCs) as counter electrodes. The catalytic activity of the CuInS(2) films based on different precursor concentrations was investigated using electrochemical methods. DSSCs based on optimized CuInS(2) thin film as counter electrodes reached a power conversion efficiency of 6.33%, comparable to that of sputtering Pt (6.07%). PMID:23388681

Yang, Jie; Bao, Chunxiong; Zhang, Jiyuan; Yu, Tao; Huang, Huan; Wei, Yulong; Gao, Hao; Fu, Gao; Liu, Jianguo; Zou, Zhigang

2013-03-11

362

Production and characterization of high-titer serum-free cell culture grown hepatitis C virus particles of genotype 1-6.  

PubMed

Recently, cell culture systems producing hepatitis C virus particles (HCVcc) were developed. Establishment of serum-free culture conditions is expected to facilitate development of a whole-virus inactivated HCV vaccine. We describe generation of genotype 1-6 serum-free HCVcc (sf-HCVcc) from Huh7.5 hepatoma cells cultured in adenovirus expression medium. Compared to HCVcc, sf-HCVcc showed 0.6-2.1log10 higher infectivity titers (4.7-6.2log10Focus Forming Units/mL), possibly due to increased release and specific infectivity of sf-HCVcc. In contrast to HCVcc, sf-HCVcc had a homogeneous single-peak density profile. Entry of sf-HCVcc depended on HCV co-receptors CD81, LDLr, and SR-BI, and clathrin-mediated endocytosis. HCVcc and sf-HCVcc were neutralized similarly by chronic-phase patient sera and by human monoclonal antibodies targeting conformational epitopes. Thus, we developed serum-free culture systems producing high-titer single-density sf-HCVcc, showing similar biological properties as HCVcc. This methodology has the potential to advance HCV vaccine development and to facilitate biophysical studies of HCV. PMID:24928051

Mathiesen, Christian K; Jensen, Tanja B; Prentoe, Jannick; Krarup, Henrik; Nicosia, Alfredo; Law, Mansun; Bukh, Jens; Gottwein, Judith M

2014-06-01

363

Detection of the quantity of kinesin and microgravity-sensitive kinesin genes in rat bone marrow stromal cells grown in a simulated microgravity environment  

NASA Astrophysics Data System (ADS)

Kinesin and kinesin-like proteins (KLPs) constitute a superfamily of microtubule motor proteins found in all eukaryotic organisms. Members of the kinesin superfamily are known to play important roles in many fundamental cellular and developmental processes. To date, few published studies have reported on the effects of microgravity on kinesin expression. In this paper, we describe the expression pattern and microgravity-sensitive genes of kinesin in rat bone marrow stromal cells cultured in a ground-based rotating bioreactor. The quantity of kinesin under the clinorotation condition was examined by immunoblot analysis with anti-kinesin. Furthermore, the distribution of kinesin at various times during clinorotation was determined by dual immunostaining, using anti-kinesin monoclonal antibody or anti-?-tubulin monoclonal antibody. In terms of kinesin quantity, we found that the ratios of the amounts of clinorotated/stationary KLPs decreased from clinorotation day 5 to day 10, although it increased on days 2 and 3. Immunofluorescence analysis revealed that kinesin in the nucleus was the first to be affected by simulated microgravity, following the kinesin at the periphery that was affected at various times during clinorotation. Real-time RT-PCR analysis of kinesin mRNA expression was performed and led to the identification of 3 microgravity-sensitive kinesin genes: KIF9, KIFC1, and KIF21A. Our results suggest that kinesin has a distinct expression pattern, and the identification of microgravity-sensitive kinesin genes offers insight into fundamental cell biology.

Ni, Chengzhi; Wang, Chunyan; Li, Yuan; Li, Yinghui; Dai, Zhongquan; Zhao, Dongming; Sun, Hongyi; Wu, Bin

2011-06-01

364

In vitro cytotoxic activity of seed oil of fenugreek against various cancer cell lines.  

PubMed

In the present study, investigations were carried out to screen the anticancer activities of fenugreek seed oil against cancer cell lines (HEp-2, MCF-7, WISH cells), and a normal cell line (Vero cells). Cytotoxicity was assessed with MTT and NRU assays, and cellular morphological alterations were studied using phase contrast light microscopy. All cells were exposed toi 10-1000 ?g/ml of fenugreek seed oil for 24 h. The results show that fenugreek seed oil significantly reduced the cell viability, and altered the cellular morphology in a dose dependent manner. Among the cell lines, HEp-2 cells showed the highest decrease in cell viability, followed by MCF-7, WISH, and Vero cells by MTT and NRU assays. Cell viability at 1000 ?g/ml was recorded as 55% in HEp-2 cells, 67% in MCF-7 cells, 75% in WISH cells, and 86% in Vero cells. The present study provides preliminary screening data for fenugreek seed oil pointing to potent cytotoxicity against cancer cells. PMID:23679282

Al-Oqail, Mai Mohammad; Farshori, Nida Nayyar; Al-Sheddi, Ebtesam Saad; Musarrat, Javed; Al-Khedhairy, Abdulaziz Ali; Siddiqui, Maqsood Ahmed

2013-01-01

365

The initiation of neurite outgrowth by sympathetic neurons grown in vitro does not depend on assembly of microtubules [published erratum appears in J Cell Biol 1995 Feb;128(3):443  

PubMed Central

Neurite formation by dissociated chick sympathetic neurons in vitro begins when one of the many filopodia that emanate from the cell body of a neuron is invaded by cytoplasm containing microtubules and other components of axoplasm (Smith, 1994). This study was undertaken to determine whether this process depends on assembly of microtubules. To inhibit microtubule assembly, neurons were grown in medium containing nocodazole or colchicine. In one series of experiments, neurons first were exposed to the microtubule-stabilizing drug, taxol, so that existing microtubules would remain intact while assembly of new microtubules was inhibited. The ability of neurons to form neurites was assessed by time-lapse video microscopy. Neurons subsequently were stained with antibodies against the tyrosinated and acetylated forms of alpha-tubulin and examined by laser confocal microscopy to visualize microtubules. Neurons were able to form short processes despite inhibition of microtubule assembly and they did so in a way that closely resembled process formation in control medium. Processes formed by neurons that had not been pretreated with taxol were devoid of microtubules. However, microtubules were present in processes of taxol- pretreated neurons. These microtubules contained acetylated alpha- tubulin, as is typical of stable microtubules, but not tyrosinated alpha-tubulin, the form present in recently assembled microtubules. These findings show that the initial steps in neurite formation do not depend on microtubule assembly and suggest that microtubules assembled in the cell body can be translocated into developing neurites as they emerge. The results are compatible with models of neurite formation which postulate that cytoplasm from the cell body is transported into filopodia by actomyosin-based motility mechanisms.

1994-01-01

366

Parameters in three-dimensional osteospheroids of telomerized human mesenchymal (stromal) stem cells grown on osteoconductive scaffolds that predict in vivo bone-forming potential.  

PubMed

Osteoblastic differentiation of human mesenchymal stem cells (hMSC) in monolayer culture is artefactual, lacking an organized bone-like matrix. We present a highly reproducible microwell protocol generating three-dimensional ex vivo multicellular aggregates of telomerized hMSC (hMSC-telomerase reverse transcriptase (TERT)) with improved mimicry of in vivo tissue-engineered bone. In osteogenic induction medium the hMSC were transitioned with time-dependent specification toward the osteoblastic lineage characterized by production of alkaline phosphatase, type I collagen, osteonectin, and osteocalcin. Introducing a 1-2 mm(3) crystalline hydroxyapatite/beta-tricalcium phosphate scaffold generated osteospheroids with upregulated gene expression of transcription factors RUNX2/CBFA1, Msx-2, and Dlx-5. An organized lamellar bone-like collagen matrix, evident by birefringence of polarized light, was deposited in the scaffold concavities. Here, mature osteoblasts stained positively for differentiated osteoblast markers TAZ, biglycan, osteocalcin, and phospho-AKT. Quantification of collagen birefringence and relatively high expression of genes for matrix proteins, including type I collagen, biglycan, decorin, lumican, elastin, microfibrillar-associated proteins (MFAP2 and MFAP5), periostin, and tetranectin, in vitro correlated predictively with in vivo bone formation. The three-dimensional hMSC-TERT/hydroxyapatite-tricalcium phosphate osteospheroid cultures in osteogenic induction medium recapitulated many characteristics of in vivo bone formation, providing a highly reproducible and resourceful platform for improved in vitro modeling of osteogenesis and refinement of bone tissue engineering. PMID:20196644

Burns, Jorge S; Rasmussen, Pernille L; Larsen, Kenneth H; Schrøder, Henrik Daa; Kassem, Moustapha

2010-07-01

367

Correlation between Phylogroups and Intracellular Proteomes of Propionibacterium acnes and Differences in the Protein Expression Profiles between Anaerobically and Aerobically Grown Cells  

PubMed Central

Propionibacterium acnes is one of the dominant commensals on the human skin and also an opportunistic pathogen in relation to acne, sarcoidosis, prostate cancer, and various infections. Recent investigations using housekeeping and virulence genes have revealed that the species consists of three major evolutionary clades (types I, II, and III). In order to investigate protein expression differences between these phylogroups, proteomic profiles of 21 strains of P. acnes were investigated. The proteins extracted from cells cultured under anaerobic and aerobic conditions were analysed using a SELDI-TOF mass spectrometer, high-resolution capillary gel electrophoresis, and LC-MS/ MS. The SELDI spectral profiles were visualised as a heat map and a dendrogram, which resulted in four proteomic groups. Strains belonging to type I were represented in the proteome Group A, while Group B contained type III strains. Groups C and D contained mixtures of types I and II. Each of these groups was not influenced by differences in culture conditions. Under anoxic growth conditions, a type IB strain yielded high expressions of some proteins, such as methylmalonyl-CoA epimerase and the Christie-Atkins-Munch-Petersen (CAMP) factor. The present study revealed good congruence between genomic and proteomic data suggesting that the microenvironment of each subtype may influence protein expression.

Dekio, Itaru; Culak, Renata; Ball, Graham; Gharbia, Saheer; Shah, Haroun N.

2013-01-01

368

Effects of V/III Ratio on the Properties of In1-xGaxP/GaAs Grown by a Valved Phosphorus Cracker Cell in Solid Source Molecular Beam Epitaxy  

NASA Astrophysics Data System (ADS)

We report the molecular beam epitaxial (MBE) growth ofhigh-quality In1-xGaxP epilayers grown on a GaAs (001)substrate using a valved phosphorus cracker cell at a wide range ofV/III flux ratios. Film characterization was carried out using X-raydiffraction (XRD), Raman scattering and low-temperaturephotoluminescence (PL). A typical Raman spectrum showed featurescharacteristic of atomic disorder in the material with theappearance of GaP-like and InP-like longitudinal-optic (LO) modesand the transverse-optic (TO) mode. The PL peak energy increasedfrom 1.941 eV to 1.967 eV while the PL full-width at half maximum(FWHM) decreased from ˜11.3 meV to ˜6.3 meV as theV/III ratio was increased from 5 to 50. This suggests an increase inthe atomic disorder (more random) and improvement in the opticalquality. The Stokes shift, estimated from the energy differencebetween the band-gap calculated using XRD composition and PL peakenergy, suggests an increase in microclustering following adecrease in the V/III ratio.

Yoon, Soon; Mah, Kia; Zheng, Hai

1999-10-01

369

An animal component free medium that promotes the growth of various animal cell lines for the production of viral vaccines.  

PubMed

IPT-AFM is a proprietary animal component free medium that was developed for rabies virus (strain LP 2061) production in Vero cells. In the present work, we demonstrated the versatility of this medium and its ability to sustain the growth of other cell lines and different virus strains. Here, three models were presented: Vero cells/rabies virus (strain LP 2061), MRC-5 cells/measles virus (strain AIK-C) and BHK-21 cells/rabies virus (strain PV-BHK21). The cell lines were first adapted to grow in IPT-AFM, by progressive reduction of the amount of serum in the culture medium. After their adaptation, BHK-21 cells grew in suspension by forming clumps, whereas MRC-5 cells remained adherent. Then, kinetics of cell growth were studied in agitated cultures for both cell lines. In addition, kinetics of virus replication were investigated. PMID:24583007

Rourou, Samia; Ben Ayed, Yousr; Trabelsi, Khaled; Majoul, Samy; Kallel, Héla

2014-05-19

370

Characterization of silicon crystals grown by the heat exchanger method  

SciTech Connect

Silicon ingots grown by the Heat Exchanger Method (HEM) as large as 45 kg in mass (34 cm x 34 cm x 17 cm) are characterized electrically and structurally. The defect state in the crystal is related to the solar cell efficiency. Such characterization indicates that the solar cell efficiency of HEM crystals is limited by the crystal perfection, but that HEM silicon has the potential to yield silicon with quality comparable to Cz grown silicon. A new approach to grow HEM material of better quality is discussed.

Hyland, S.; Dumas, K.A.; Engelbrecht, J.A.A.; Leung, D.; Schwuttke, G.M.

1983-05-01

371

An international outbreak of Vero cytotoxin-producing Escherichia coli O157 infection amongst tourists; a challenge for the European infectious disease surveillance network.  

PubMed

In March 1997, an outbreak of Vero cytotoxin-producing Escherichia coli O157 (VTEC) infection occurred amongst holidaymakers returning from Fuerteventura, Canary Islands. For the investigation, a confirmed case was an individual staying in Fuerteventura during March 1997, with either E. coli O157 VTEC isolated in stool, HUS or serological evidence of recent infection; a probable case was an individual with bloody diarrhoea without laboratory confirmation. Local and Europe-wide active case finding was undertaken through national centres, Salm-Net and the European Programme of Intervention Epidemiology, followed by a case-control study. Fourteen confirmed and one probable case were identified from England (7), Finland (5), Wales (1), Sweden (1) and Denmark (1) staying in four hotels. Three of the four hotels were supplied with water from a private well which appeared to be the probable vehicle of transmission. The case-control study showed illness was associated with consumption of raw vegetables (OR 8.4, 95% CI 1-5-48.2) which may have been washed in well water. This investigation shows the importance of international collaboration in the detection and investigation of clusters of enteric infection. PMID:10579440

Pebody, R G; Furtado, C; Rojas, A; McCarthy, N; Nylen, G; Ruutu, P; Leino, T; Chalmers, R; de Jong, B; Donnelly, M; Fisher, I; Gilham, C; Graverson, L; Cheasty, T; Willshaw, G; Navarro, M; Salmon, R; Leinikki, P; Wall, P; Bartlett, C

1999-10-01

372

Protein Crystals Grown in Space  

NASA Technical Reports Server (NTRS)

A collage of protein and virus crystals, many of which were grown on the U.S. Space Shuttle or Russian Space Station, Mir. The crystals include the proteins canavalin; mouse monoclonal antibody; a sweet protein, thaumatin; and a fungal protease. Viruses are represented here by crystals of turnip yellow mosaic virus and satellite tobacco mosaic virus. The crystals are photographed under polarized light (thus causing the colors) and range in size from a few hundred microns in edge length up to more than a millimeter. All the crystals are grown from aqueous solutions and are useful for X-ray diffraction analysis. Credit: Dr. Alex McPherson, University of California, Irvine.

2000-01-01

373

Induction of micronuclei by Zearalenone in Vero monkey kidney cells and in bone marrow cells of mice: protective effect of Vitamin E  

Microsoft Academic Search

Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin mainly produced by Fusarium graminaerum, found as a world-wide contaminant mainly of corn and wheat. Previous studies have demonstrated that among several other effects on animals and humans, ZEN also displays hepatotoxicity, immunotoxicity and nephrotoxicity. ZEN is mainly known as a hormonal disrupter due to its estrogenic activities and consequent toxicity for reproduction.

Zouhour Ouanes; Salwa Abid; Imen Ayed; Rachid Anane; Théophile Mobio; Edmond E Creppy; Hassen Bacha

2003-01-01

374

DNA fragmentation, apoptosis and cell cycle arrest induced by zearalenone in cultured DOK, Vero and Caco-2 cells: prevention by Vitamin E  

Microsoft Academic Search

Zearalenone (ZEN) is a non-steroidal oestrogenic mycotoxin produced by several Fusarium species growing on cereals. ZEN and its metabolites bind to human oestrogen receptors and hence display oestrogenic and anabolic properties. Several lines of investigation suggest that ZEN may be genotoxic in vivo. ZEN damages DNA in Bacillus subtilis recombination tests, and it induces sister chromatid exchange and chromosomal aberration

Salwa Abid-Essefi; Isabelle Baudrimont; Wafa Hassen; Zouhour Ouanes; Théophile A. Mobio; Rachid Anane; Edmond E. Creppy; Hassen Bacha

2003-01-01

375

Diverse apoptotic pathways in enterovirus 71-infected cells  

Microsoft Academic Search

Mechanisms related to the neuropathogenesis of enterovirus 71 infection remain unclear. This investigation conducts a comprehensive\\u000a study of the apoptotic pathways in neural and non-neural cells following enterovirus 71 infection. Infections with enterovirus\\u000a 71 not only induce classical cytopathic effects in SF268 (human glioblastoma), SK-N-MC (human neuroblastoma), RD, and Vero\\u000a cells, but also induce classic signs of apoptosis in all

Shih-Cheng Chang; Jing-Yi Lin; Lily Yen-Cheng Lo; Mei-Ling Li; Shin-Ru Shih

2004-01-01

376

Cytopathogenesis of Naegleria fowleri Thai strains for cultured human neuroblastoma cells.  

PubMed

The aim of this study is to evaluate cellular interaction between free-living amoebae Naegleria fowleri strains and mammalian target cells in vitro. Two Thai strains of N. fowleri; Khon Kaen strain from the environment and Siriraj strain from the patient's cerebrospinal fluid and the Center of Disease Control VO 3081 strain from Atlanta (US) were studied. Human neuroblastoma (SK-N-MC) and African Green monkey Kidney (Vero) cells were used as target cells. Each cell line was inoculated with each strain of N. fowleri at a ratio of 1:1 and observed for 7 days. The uninoculated target cells and each strain of N. fowleri were used as control. The numbers of the challenged and unchallenged cells as well as the free-living amoebae were counted three times by trypan blue exclusion method. The inoculation began when the amoebae attached to the cell membrane and ingested the target cells. In this study, extensive cytopathogenesis with many floating inoculated cells and abundant number of amoebae were observed. The destruction pattern of both inoculated SK-N-MC and Vero target cells were similar. Interestingly, SK-N-MC was more susceptible to N. fowleri strains than the Vero cell. In addition, N. fowleri Siriraj strain showed the highest destruction pattern for each target cell. Our findings suggest that the SK-N-MC should be used as a base model for studying the neuropathogenesis in primary amoebic meningoencephalitis patients. PMID:18214541

Tiewcharoen, Supathra; Malainual, Nat; Junnu, Virach; Chetanachan, Pruksawan; Rabablert, Jundee

2008-04-01

377

Extremely low-frequency electromagnetic fields cause DNA strand breaks in normal cells  

PubMed Central

Background Extremely low frequency electromagnetic fields aren’t considered as a real carcinogenic agent despite the fact that some studies have showed impairment of the DNA integrity in different cells lines. The aim of this study was evaluation of the late effects of a 100 Hz and 5.6 mT electromagnetic field, applied continuously or discontinuously, on the DNA integrity of Vero cells assessed by alkaline Comet assay and by cell cycle analysis. Normal Vero cells were exposed to extremely low frequency electromagnetic fields (100 Hz, 5.6 mT) for 45 minutes. The Comet assay and cell cycle analysis were performed 48 hours after the treatment. Results Exposed samples presented an increase of the number of cells with high damaged DNA as compared with non-exposed cells. Quantitative evaluation of the comet assay showed a significantly (<0.001) increase of the tail lengths, of the quantity of DNA in tail and of Olive tail moments, respectively. Cell cycle analysis showed an increase of the frequency of the cells in S phase, proving the occurrence of single strand breaks. The most probable mechanism of induction of the registered effects is the production of different types of reactive oxygen species. Conclusions The analysis of the registered comet indices and of cell cycle showed that extremely low frequency electromagnetic field of 100 Hz and 5.6 mT had a genotoxic impact on Vero cells.

2014-01-01

378

Tissue grown in space in NASA Bioreactor  

NASA Technical Reports Server (NTRS)

Dr. Lisa E. Freed of the Massachusetts Institute of Technology and her colleagues have reported that initially disc-like specimens tend to become spherical in space, demonstrating that tissues can grow and differentiate into distinct structures in microgravity. The Mir Increment 3 (Sept. 16, 1996 - Jan. 22, 1997) samples were smaller, more spherical, and mechanically weaker than Earth-grown control samples. These results demonstrate the feasibility of microgravity tissue engineering and may have implications for long human space voyages and for treating musculoskeletal disorders on earth. Final samples from Mir and Earth appeared histologically cartilaginous throughout their entire cross sections (5-8 mm thick), with the exception of fibrous outer capsules. Constructs grown on Earth (A) appeared to have a more organized extracellular matrix with more uniform collagen orientation as compared with constructs grown on Mir (B), but the average collagen fiber diameter was similar in the two groups (22 +- 2 nm) and comparable to that previously reported for developing articular cartilage. Randomly oriented collagen in Mir samples would be consistent with previous reports that microgravity disrupts fibrillogenesis. These are transmission electron micrographs of constructs from Mir (A) and Earth (B) groups at magnifications of x3,500 and x120,000 (Inset). The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Credit: Proceedings of the National Academy of Sciences.

2001-01-01

379

Tissue grown in space in NASA Bioreactor  

NASA Technical Reports Server (NTRS)

For 5 days on the STS-70 mission, a bioreactor cultivated human colon cancer cells, such as the culture section shown here, which grew to 30 times the volume of control specimens grown on Earth. This significant result was reproduced on STS-85 which grew mature structures that more closely match what are found in tumors in humans. The two white circles within the tumor are part of a plastic lattice that helped the cells associate. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

1998-01-01

380

Apoptosis as a Cause of Death in Measles Virus-Infected Cells  

Microsoft Academic Search

To determine the mechanism of measles virus-induced cell death, we studied the infection of Vero cells and monocytic cell lines with wild-type (Chicago-1) and vaccine (Edmonston) strains of measles virus. DNA fragmentationindicativeofapoptosiswasapparentbyflowcytometry,agarosegelelectrophoresis,andelectron microscopy. Within syncytia, DNA strand breaks were demonstrated by end labeling with terminal transferase and then by visualization. A number of viruses have recently been shown to cause

LISA M. ESOLEN; SUK W. PARK; J. MARIE HARDWICK; ANDDIANE E. GRIFFIN

1995-01-01

381

Kraft Pulping of Coppice Grown Eucalypts.  

National Technical Information Service (NTIS)

Pulping and bleaching investigations were carried out on coppice grown Eucalyptus tereticornis and Eucalyptus grandis. It was observed that bleachable grade pulps could be obtained from coppice grown eucalypts. These pulps could be bleached to 76-78% ISO ...

R. Pant A. K. Rai R. M. Mathur Y. V. Sood

1980-01-01

382

Dielectric properties of rapidly grown KDP crystals  

Microsoft Academic Search

The dielectric properties of rapidly grown potassium dihydrogen phosphate KH2PO4 (KDP) crystals have been studied over a wide temperature range and compared with the properties of traditionally grown KDP\\u000a crystals. It was found that the contribution of domains to permittivity in rapidly grown crystals is considerably less than\\u000a in conventionally grown ones. The dielectric properties in various growth sectors of

S. V. Grabovskii; I. V. Shnaidshtein; B. A. Strukov

2003-01-01

383

Fast Plants Grown in Light and Dark  

NSDL National Science Digital Library

Photograph of two five-day-old Standard Fast Plants grown in Bottle Growing Systems--one grown with full light, one grown in the dark. This is a good example of a quick way to stimulate discussion about the matter and energy sources and needs that germinating seeds have in comparison to seedlings or plants.

Lauffer, Hedi B.

384

Light effects in yeast: inhibition by visible light of growth and transport in Saccharomyces cerevisiae grown at low temperatures.  

PubMed Central

Growth rate, sugar transport, and amino acid transport of yeast cells grown at 12 degrees C were inhibited by cool-white fluorescent light. At light intensities below 1,250 lx, growth and membrane transport were only slightly inhibited. Above 1,250 lx, there was increasing inhibition of both processes. Transport of histidine was completely inhibited after 3 to 5 days in cultures grown at 12 degrees C under 3,500-lx illumination. Cells grown at 20 degrees C were not inhibited by light intensities that caused complete loss of viability and membrane transport activity in cells grown at 12 degrees C.

Woodward, J R; Cirillo, V P; Edmunds, L N

1978-01-01

385

Apoptosis in wheat seedlings grown under normal daylight.  

PubMed

Apoptosis was observed in the coleoptile and initial leaf in 5-8-day-old wheat seedlings grown under normal daylight. Apoptosis is an obligatory event in early wheat plant ontogenesis, and it is characterized by cytoplasmic structural reorganization and fragmentation, in particular, with the appearance in vacuoles of specific vesicles containing intact organelles, chromatin condensation and margination in the nucleus, and internucleosomal fragmentation of nuclear DNA. The earliest signs of programmed cell death (PCD) were observed in the cytoplasm, but the elements of apoptotic degradation in the nucleus appeared later. Nuclear DNA fragmentation was detected after chromatin condensation and the appearance in vacuoles of specific vesicles containing mitochondria. Two PCD varieties were observed in the initial leaf of 5-day-old seedlings grown under normal daylight: a proper apoptosis and vacuolar collapse. On the contrary, PCD in coleoptiles under various growing (light) conditions and in the initial leaf of etiolated seedlings is only a classical plant apoptosis. Therefore, various tissue-specific and light-dependent PCD forms do exist in plants. Amounts of O2*- and H2O2 evolved by seedlings grown under normal daylight are less than that evolved by etiolated seedlings. The amount of H2O2 formed in the presence of sodium salicylate or azide by seedlings grown under normal daylight was increased. Contrary to etiolated seedlings, the antioxidant BHT (ionol) did not inhibit O2*- formation and apoptosis and it had no influence on ontogenesis in the seedlings grown under normal daylight. Thus, in plants grown under the normal light regime the powerful system controlling the balance between formation and inactivation of reactive oxygen species (ROS) does exist and it effectively functions. This system is responsible for maintenance of cell homeostasis, and it regulates the crucial ROS level controlling plant growth and development. In etiolated plants, this system seems to be absent, or it is much less effective. PMID:15061695

Aleksandrushkina, N I; Zamyatnina, V A; Bakeeva, L E; Seredina, A V; Smirnova, E G; Yaguzhinsky, L S; Vanyushin, B F

2004-03-01