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181.
One of the limitations of the use of phage antibody libraries in high throughput selections is the production of sufficient phage antibody library at the appropriate quality. Here, we successfully adapt a bioreactor-based protocol for the production of phage peptide libraries to the production of phage antibody libraries. The titers obtained in the stirred-tank bioreactor are 4 to 5 times higher than in a standard shake flask procedure, and the quality of the phage antibody library produced is indistinguishable to that produced using standard procedures as assessed by Western blotting and functional selections. Availability of this protocol will facilitate the use of phage antibody libraries in high-throughput scale selections. 相似文献
182.
D Deprez J Fransen M Lenoir RM Philippaerts R Vaeyens 《Biology of sport / Institute of Sport》2015,32(1):65-70
The aim of the study was to investigate test reliability of the Yo-Yo intermittent recovery test level 1 (YYIR1) in 36 high-level youth soccer players, aged between 13 and 18 years. Players were divided into three age groups (U15, U17 and U19) and completed three YYIR1 in three consecutive weeks. Pairwise comparisons were used to investigate test reliability (for distances and heart rate responses) using technical error (TE), coefficient of variation (CV), intra-class correlation (ICC) and limits of agreement (LOA) with Bland-Altman plots. The mean YYIR1 distances for the U15, U17 and U19 groups were 2024 ± 470 m, 2404 ± 347 m and 2547 ± 337 m, respectively. The results revealed that the TEs varied between 74 and 172 m, CVs between 3.0 and 7.5%, and ICCs between 0.87 and 0.95 across all age groups for the YYIR1 distance. For heart rate responses, the TEs varied between 1 and 6 bpm, CVs between 0.7 and 4.8%, and ICCs between 0.73 and 0.97. The small ratio LOA revealed that any two YYIR1 performances in one week will not differ by more than 9 to 28% due to measurement error. In summary, the YYIR1 performance and the physiological responses have proven to be highly reliable in a sample of Belgian high-level youth soccer players, aged between 13 and 18 years. The demonstrated high level of intermittent endurance capacity in all age groups may be used for comparison of other prospective young soccer players. 相似文献
183.
Persistent Infection of Rhesus Macaques by the Rev-Independent Nef(−) Simian Immunodeficiency Virus SIVmac239: Replication Kinetics and Genomic Stability
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Agneta S. von Gegerfelt Vladimir Liska Nancy B. Ray Harold M. McClure Ruth M. Ruprecht Barbara K. Felber 《Journal of virology》1999,73(7):6159-6165
We generated previously a Nef(-), replication-competent clone of SIVmac239 in which the Rev protein and the Rev-responsive element were replaced by the constitutive transport element (CTE) of simian retrovirus type 1 (A. S. von Gegerfelt and B. K. Felber, Virology 232:291-299, 1997). In the present report, we show that this virus was able to infect and replicate in rhesus macaques. The Rev-independent Nef(-) simian immunodeficiency virus induced a persistent humoral immune response in all monkeys, although viral loads were very low. Upon propagation in the monkeys, the genotype remained stable and the virus retained its in vitro growth characteristics. The infected monkeys showed normal hematological values and no signs of disease at more than 18 months post-virus exposure. Therefore, replacement of the essential Rev regulation by the CTE generated a virus variant that retained its replicative capacity both in vitro and in vivo, albeit at low levels. 相似文献
184.
Tissue selectivity of murine leukemia virus infection is determined by long terminal repeat sequences. 总被引:45,自引:34,他引:11
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Polyomavirus mutants were isolated from PCC4 embryonal carcinoma cells infected with a variant strain of polyomavirus (ev 1001h) and were found to contain a tandem duplication overlapping the enhancers and the origin of replication. These mutants were able to infect several lines of embryonal carcinoma cells, including PCC4, F9, and LT1. The sequence and structure of one of these mutants are presented and compared with those of other PyEC PCC4 mutants previously described. 相似文献
185.
Summary Chromosome studies were performed on peripheral lymphocytes from two patients with Werner syndrome and two healthy control individuals to detect spontaneous and/or mutagen-induced chromosomal instability of this disease. Diepoxybutane, isonicotinic acid hydrazide, 4-nitro-quinoline-1-oxide, and bleomycin were used as standard clastogens. While the spontaneous frequency of chromosomal breakage was much higher in lymphocytes from both patients than in the control cells, the basic rate of sister chromatid exchange (SCE) was found to be in the control range. The sensitivity to clastogens of the patients' cells, however, was not substantially increased as compared with the controls if the degree of multiplication of the spontaneous breakage rate or SCE frequency was taken as the basis for comparison. No indication of a greater inhibition of proliferation by the clastogens in the patients' cells than in normal cells was observed using BrdU-labelled lymphocytes. Thus, the lymphocytes from both patients of the present study lacked essential features of the classical chromosome instability syndromes. 相似文献
186.
Using inbreeding theory as applied to neutral alleles inherited maternally,
we generate expected probability distributions of times to identity by
descent for random pairs of mitochondrial genotypes within a population or
within an entire species characterized by high gene flow. For comparisons
with these expectations, empirical distributions of times to most recent
common ancestry were calculated (by conventional mtDNA clock calibrations)
from mtDNA haplotype distances observed within each of three vertebrate
species--American eels, hardhead catfish, and redwinged blackbirds. These
species were chosen for analysis because census population size in each is
currently large and because both genetic and life-history data are
consistent with the postulate that historical gene flow within these
species has been high. The observed molecular distances among mtDNA
lineages were two to three orders of magnitude lower than predicted from
census sizes of breeding females, suggesting that rate of mtDNA evolution
is decelerated in these species and/or that long-term effective population
size is vastly smaller than present-day population size. Several
considerations point to the latter possibility as most likely. The genetic
structure of any species is greatly influenced by historical demography;
even for species that are currently abundant, mtDNA gene lineages appear to
have been channeled through fairly small numbers of ancestors.
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187.
Developmental and wound-, cold-, desiccation-, ultraviolet-B-stress-induced modulations in the expression of the petunia zinc finger transcription factor gene ZPT2-2
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van Der Krol AR van Poecke RM Vorst OF Voogt C van Leeuwen W Borst-Vrensen TW Takatsuji H van Der Plas LH 《Plant physiology》1999,121(4):1153-1162
The ZPT2-2 gene belongs to the EPF gene family in petunia (Petunia hybrida), which encodes proteins with TFIIIA-type zinc-finger DNA-binding motifs. To elucidate a possible function for ZPT2-2, we analyzed its pattern of expression in relation to different developmental and physiological stress signals. The activity of the ZPT2-2 promoter was analyzed using a firefly luciferase (LUC) reporter gene, allowing for continuous measurements of transgene activity in planta. We show that ZPT2-2::LUC is active in all plant tissues, but is strongly modulated in cotyledons upon germination, in leaves in response to desiccation, cold treatment, wounding, or ultraviolet-B light, and in petal tissue in response to pollination of the stigma. Analysis of mRNA levels indicated that the modulations in ZPT2-2::LUC expression reflect modulations in endogenous ZPT2-2 gene expression. The change in ZPT2-2::LUC activity by cold treatment, wounding, desiccation, and ultraviolet-B light suggest that the phytohormones ethylene and jasmonic acid are involved in regulating the expression of ZPT2-2. Although up-regulation of expression of ZPT2-2 can be blocked by inhibitors of ethylene perception, expression in plants is not induced by exogenously applied ethylene. The application of jasmonic acid does result in an up-regulation of gene activity and, thus, ZPT2-2 may play a role in the realization of the jasmonic acid hormonal responses in petunia. 相似文献
188.
Fredrik RM Huss Erika Nyman Carl-Johan Gustafson Katrin Gisself?lt Elisabeth Liljensten Gunnar Kratz 《Organogenesis》2008,4(3):195-200
Full thickness skin wounds in humans heal with scars, but without regeneration of the dermis. A degradable poly(urethane urea) scaffold (PUUR), Artelon® is already used to reinforce soft tissues in orthopaedics, and for treatment of osteoarthritis of the hand, wrist and foot. In this paper we have done in vitro experiments followed by in vivo studies to find out whether the PUUR is biocompatible and usable as a template for dermal regeneration. Human dermal fibroblasts were cultured on discs of PUUR, with different macrostructures (fibrous and porous). They adhered to and migrated into the scaffolds, and produced collagen. The porous scaffold was judged more suitable for clinical applications and 4 mm Ø, 2 mm-thick discs of porous scaffold (12% w/w or 9% w/w polymer solution) were inserted intradermally in four healthy human volunteers. The implants were well tolerated and increasing ingrowth of fibroblasts was seen over time in all subjects. The fibroblasts stained immunohistochemically for procollagen and von Willebrand factor, indicating neocollagenesis and angiogenesis within the scaffolds. The PUUR scaffold may be a suitable material to use as a template for dermal regeneration.Key words: dermal regeneration, tissue engineering, polymer scaffold, wound healing, in vitro, in vivo, guided tissue regeneration, human, burns 相似文献
189.
Ruprecht K Ferreira H Flockerzi A Wahl S Sauter M Mayer J Mueller-Lantzsch N 《Journal of virology》2008,82(20):10008-10016
190.
A higher‐level classification of the Pannonian and western Pontic steppe grasslands (Central and Eastern Europe)
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Wolfgang Willner Anna Kuzemko Jürgen Dengler Milan Chytrý Norbert Bauer Thomas Becker Claudia Biţă‐Nicolae Zoltán Botta‐Dukát Andraž Čarni János Csiky Ruzica Igić Zygmunt Kącki Iryna Korotchenko Matthias Kropf Mirjana Krstivojević‐Ćuk Daniel Krstonošić Tamás Rédei Eszter Ruprecht Luise Schratt‐Ehrendorfer Yuri Semenishchenkov Zvjezdana Stančić Yulia Vashenyak Denys Vynokurov Monika Janišová 《应用植被学》2017,20(1):143-158