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51.
To meet the increasing requirement for therapeutic antibodies to conduct clinical trials, an enhanced culture medium and fed-batch process was developed for GS-NS0 cell lines. This process was shown to produce high concentrations of monoclonal antibodies for several cell lines expressing different antibodies. Cells were adapted to growth in a glutamine- and serum-free medium containing bovine serum albumin (BSA), cholesterol, and transferrin. A number of amino acids were found to be depleted during cell culture. The concentrations of these amino acids were increased, and further cell culture analyses were performed. This process of cell growth and analysis was repeated over multiple cycles until no depletion was detected. This resulted in an amino acid supplement that was shown to be generic and enhanced antibody productivity up to 5-fold for the three cell lines tested. Transferrin was replaced using tropolone, a lipophilic iron chelator and ferric ammonium citrate. Cell growth was equivalent to that in transferrin-containing medium over the wide ranges tested. A concentrated feed solution, based on the amino acid supplement and the components of the serum- and protein-free supplements, was formulated. Addition of this feed in response to metabolic requirements resulted in a harvest titer a further 2-fold higher than the enhanced culture medium. Harvest antibody titers of up to 600 mg/L were achieved for three cell lines expressing different antibodies, representing an increase of 10-fold over the starting concentrations. 相似文献
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Effects of roads and land use on frog distributions across spatial scales and regions in the Eastern and Central United States 下载免费PDF全文
David M. Marsh Bradley J. Cosentino Kara S. Jones Joseph J. Apodaca Karen H. Beard Jane Margaret Bell Christine Bozarth Derrick Carper Julie F. Charbonnier Andreia Dantas Elizabeth A. Forys Miran Foster Jaquelyn General Kristen S. Genet Macie Hanneken Kyle R. Hess Shane A. Hill Faisal Iqbal Nancy E. Karraker Eran S. Kilpatrick Tom A. Langen James Langford Kathryn Lauer Alison J. McCarthy Joseph Neale Saumya Patel Austin Patton Cherie Southwick Nathaniel Stearrett Nicholas Steijn Mohammad Tasleem Joseph M. Taylor James R. Vonesh 《Diversity & distributions》2017,23(2):158-170
54.
Cheng Ly Tamar Melman Alison L. Barth G. Bard Ermentrout 《Journal of computational neuroscience》2011,30(2):211-223
BK channels are large conductance potassium channels gated by calcium and voltage. Paradoxically, blocking these channels has been shown experimentally to increase or decrease the firing rate of neurons, depending on the neural subtype and brain region. The mechanism for how this current can alter the firing rates of different neurons remains poorly understood. Using phase-resetting curve (PRC) theory, we determine when BK channels increase or decrease the firing rates in neural models. The addition of BK currents always decreases the firing rate when the PRC has only a positive region. When the PRC has a negative region (type II), BK currents can increase the firing rate. The influence of BK channels on firing rate in the presence of other conductances, such as I m and I h , as well as with different amplitudes of depolarizing input, were also investigated. These results provide a formal explanation for the apparently contradictory effects of BK channel antagonists on firing rates. 相似文献
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Peng Lin Sarah M. Hartz Zhehao Zhang Scott F. Saccone Jia Wang Jay A. Tischfield Howard J. Edenberg John R. Kramer Alison M.Goate Laura J. Bierut John P. Rice for the COGA Collaborators COGEND Collaborators GENEVA 《PloS one》2010,5(3)
Background
As the amount of data from genome wide association studies grows dramatically, many interesting scientific questions require imputation to combine or expand datasets. However, there are two situations for which imputation has been problematic: (1) polymorphisms with low minor allele frequency (MAF), and (2) datasets where subjects are genotyped on different platforms. Traditional measures of imputation cannot effectively address these problems.Methodology/Principal Findings
We introduce a new statistic, the imputation quality score (IQS). In order to differentiate between well-imputed and poorly-imputed single nucleotide polymorphisms (SNPs), IQS adjusts the concordance between imputed and genotyped SNPs for chance. We first evaluated IQS in relation to minor allele frequency. Using a sample of subjects genotyped on the Illumina 1 M array, we extracted those SNPs that were also on the Illumina 550 K array and imputed them to the full set of the 1 M SNPs. As expected, the average IQS value drops dramatically with a decrease in minor allele frequency, indicating that IQS appropriately adjusts for minor allele frequency. We then evaluated whether IQS can filter poorly-imputed SNPs in situations where cases and controls are genotyped on different platforms. Randomly dividing the data into “cases” and “controls”, we extracted the Illumina 550 K SNPs from the cases and imputed the remaining Illumina 1 M SNPs. The initial Q-Q plot for the test of association between cases and controls was grossly distorted (λ = 1.15) and had 4016 false positives, reflecting imputation error. After filtering out SNPs with IQS<0.9, the Q-Q plot was acceptable and there were no longer false positives. We then evaluated the robustness of IQS computed independently on the two halves of the data. In both European Americans and African Americans the correlation was >0.99 demonstrating that a database of IQS values from common imputations could be used as an effective filter to combine data genotyped on different platforms.Conclusions/Significance
IQS effectively differentiates well-imputed and poorly-imputed SNPs. It is particularly useful for SNPs with low minor allele frequency and when datasets are genotyped on different platforms. 相似文献57.
Identification in forensic casework by mtDNA sequencing is predominantly done by sequencing the non-coding control region (HVI and HVII). In an attempt to further increase the discriminatory power of mtDNA analysis, we sequenced a coding region between nt8306 and nt9021 to identify additional polymorphisms in a group of 61 unrelated German individuals who had mtDNA profiles that occurred more than two times each, as well as a control group of 119 unrelated Germans whose profiles occurred one or two times each. Within these 180 individuals, 38 different polymorphisms in this region were observed; 64.4% of these individuals displayed the Cambridge reference sequence profile plus A8860G. For 28 individuals with the two most common profiles, A263G-315.1insC (N = 18) and A263G-309.1insC-315.1insC (N = 10), additional polymorphisms in this coding region permitted further discrimination of 56 and 40% of the individuals, respectively. 相似文献
58.
Cave A Grieve D Johar S Zhang M Shah AM 《Philosophical transactions of the Royal Society of London. Series B, Biological sciences》2005,360(1464):2327-2334
Chronic heart failure, secondary to left ventricular hypertrophy or myocardial infarction, is a condition with increasing morbidity and mortality. Although the mechanisms underlying the development and progression of this condition remain a subject of intense interest, there is now growing evidence that redox-sensitive pathways play an important role. This article focuses on the involvement of reactive oxygen species derived from a family of superoxide-generating enzymes, termed NADPH oxidases (NOXs), in the pathophysiology of ventricular hypertrophy, the accompanying interstitial fibrosis and subsequent heart failure. In particular, the apparent ability of the different NADPH oxidase isoforms to define the response of a cell to a range of physiological and pathophysiological stimuli is reviewed. If confirmed, these data would suggest that independently targeting different members of the NOX family may hold the potential for therapeutic intervention in the treatment of cardiac disease. 相似文献
59.
Steven D. Johnson Marco G. Balducci Alison Bijl Miguel Castañeda-Zárate Ruth J. Cozien Carmen R. Ortmann Timotheüs van der Niet 《Ecological Entomology》2020,45(3):751-755
1. Diel rhythms of foraging activity by animal flower visitors can reflect niche partitioning and are considered an important component of selection on floral traits. However, it has been notoriously difficult to obtain objective information on the patterns of flower visitation by crepuscular and nocturnal insects. 2. Motion-activated cameras were used for field-based studies of hawkmoth foraging behaviour on six African plant species. 3. The results showed that short-tongued hawkmoth species forage mainly around dusk and then sporadically throughout the night, whereas long-tongued hawkmoth species feed consistently throughout the night, with a peak shortly before midnight. 4. These results provide the first quantitative estimates of diel patterns of interactions between multiple hawkmoth and plant species and, when combined with qualitative reports from other studies, suggest that differences in diel activity between the two main hawkmoth functional groups (short- and long-tongued) are consistent across the Old and New Worlds. 相似文献
60.
Claudine Porta Abhay Kotecha Alison Burman Terry Jackson Jingshan Ren Silvia Loureiro Ian M. Jones Elizabeth E. Fry David I. Stuart Bryan Charleston 《PLoS pathogens》2013,9(3)
Foot-and-mouth disease remains a major plague of livestock and outbreaks are often economically catastrophic. Current inactivated virus vaccines require expensive high containment facilities for their production and maintenance of a cold-chain for their activity. We have addressed both of these major drawbacks. Firstly we have developed methods to efficiently express recombinant empty capsids. Expression constructs aimed at lowering the levels and activity of the viral protease required for the cleavage of the capsid protein precursor were used; this enabled the synthesis of empty A-serotype capsids in eukaryotic cells at levels potentially attractive to industry using both vaccinia virus and baculovirus driven expression. Secondly we have enhanced capsid stability by incorporating a rationally designed mutation, and shown by X-ray crystallography that stabilised and wild-type empty capsids have essentially the same structure as intact virus. Cattle vaccinated with recombinant capsids showed sustained virus neutralisation titres and protection from challenge 34 weeks after immunization. This approach to vaccine antigen production has several potential advantages over current technologies by reducing production costs, eliminating the risk of infectivity and enhancing the temperature stability of the product. Similar strategies that will optimize host cell viability during expression of a foreign toxic gene and/or improve capsid stability could allow the production of safe vaccines for other pathogenic picornaviruses of humans and animals. 相似文献