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21.
Sven Göbel Karim E. Jaén Marie Dorn Victoria Neumeyer Ingo Jordan Volker Sandig Udo Reichl Jennifer Altomonte Yvonne Genzel 《Biotechnology and bioengineering》2023,120(9):2639-2657
We present a proof-of-concept study for production of a recombinant vesicular stomatitis virus (rVSV)-based fusogenic oncolytic virus (OV), rVSV-Newcastle disease virus (NDV), at high cell densities (HCD). Based on comprehensive experiments in 1 L stirred tank reactors (STRs) in batch mode, first optimization studies at HCD were carried out in semi-perfusion in small-scale cultivations using shake flasks. Further, a perfusion process was established using an acoustic settler for cell retention. Growth, production yields, and process-related impurities were evaluated for three candidate cell lines (AGE1.CR, BHK-21, HEK293SF)infected at densities ranging from 15 to 30 × 106 cells/mL. The acoustic settler allowed continuous harvesting of rVSV-NDV with high cell retention efficiencies (above 97%) and infectious virus titers (up to 2.4 × 109 TCID50/mL), more than 4–100 times higher than for optimized batch processes. No decrease in cell-specific virus yield (CSVY) was observed at HCD, regardless of the cell substrate. Taking into account the accumulated number of virions both from the harvest and bioreactor, a 15–30 fold increased volumetric virus productivity for AGE1.CR and HEK293SF was obtained compared to batch processes performed at the same scale. In contrast to all previous findings, formation of syncytia was observed at HCD for the suspension cells BHK 21 and HEK293SF. Oncolytic potency was not affected compared to production in batch mode. Overall, our study describes promising options for the establishment of perfusion processes for efficient large-scale manufacturing of fusogenic rVSV-NDV at HCD for all three candidate cell lines. 相似文献
22.
Sven Rossel Patricia Kaiser Maya Bode-Dalby Jasmin Renz Silke Laakmann Holger Auel Wilhelm Hagen Pedro Martínez Arbizu Janna Peters 《Molecular ecology resources》2023,23(2):382-395
Species identification is pivotal in biodiversity assessments and proteomic fingerprinting by MALDI-TOF mass spectrometry has already been shown to reliably identify calanoid copepods to species level. However, MALDI-TOF data may contain more information beyond mere species identification. In this study, we investigated different ontogenetic stages (copepodids C1–C6 females) of three co-occurring Calanus species from the Arctic Fram Strait, which cannot be identified to species level based on morphological characters alone. Differentiation of the three species based on mass spectrometry data was without any error. In addition, a clear stage-specific signal was detected in all species, supported by clustering approaches as well as machine learning using Random Forest. More complex mass spectra in later ontogenetic stages as well as relative intensities of certain mass peaks were found as the main drivers of stage distinction in these species. Through a dilution series, we were able to show that this did not result from the higher amount of biomass that was used in tissue processing of the larger stages. Finally, the data were tested in a simulation for application in a real biodiversity assessment by using Random Forest for stage classification of specimens absent from the training data. This resulted in a successful stage-identification rate of almost 90%, making proteomic fingerprinting a promising tool to investigate polewards shifts of Atlantic Calanus species and, in general, to assess stage compositions in biodiversity assessments of Calanoida, which can be notoriously difficult using conventional identification methods. 相似文献
23.
Within a subprogram of Integrated Monitoring (IM), understorey vegetation in Swedish natural forests was observed at fifteen reference sites over the country for twelve seasons, 1982–1993. The main task of the subprogram was to assess the impact of atmospheric deposition, mainly sulphur and nitrogen, on natural vegetation through time. The present study is focused on the variability of plant species diversity at community level and the possible impact of sulphur and nitrogen deposition. Species richness, evenness and diversity varied greatly among the sites, and between years within each site. Regarding only coniferous forests the species richness was higher in the north than in the south. But the effects of site condition and atmospheric deposition were not clarified. Changes in species diversity through time differed from site to site. No overall temporal trend was found. The atmospheric deposition of sulphur and nitrogen demonstrated a clear geographical pattern being low in the north-west and high in the south-west. Sulphur deposition declined significantly in Southern Sweden during the period. We concluded that the species diversity of understorey vegetation at the Swedish IM sites was not significantly influenced by atmospheric deposition. The changes observed are explained as natural processes. 相似文献
24.
Recent work with the green alga Dunaliella salina showed thepresence of a {small tilde}20 kDa chloroplast protein that wasrecognized by polyclonal antibodies raised against the isolatedLHC-II [Webb M.R. and Melis A. (1995) Plant Physiol. 107: 885].In this report, a characterization of the {small tilde}20 kDapolypeptide is presented. It is shown that it is localized inthe chloroplast envelope membrane of D. salina. The abundanceof this protein is constant on a per cell basis and independentof the light regime during cell growth. The {small tilde}20kDa polypeptide is easily degraded to a {small tilde}19 kDaproduct during sample preparation. A limited amino acid sequenceof 21 residues from the free N-terminus of the {small tilde}19kDa product was obtained. On the basis of this partial sequence,it was concluded that the {small tilde}20 kDa polypeptide isnot a degradation product of a known LHC-II but rather a novelprotein. The {small tilde}20kDa polypeptide did not cross-reactwith antibodies raised against the Cbr (carotene biosynthesis-related)gene product and showed a different electrophoretic mobilityfrom the latter. Light-shift experiments suggest that the {smalltilde}20 kDa polypeptide is not an ELIP (early light-inducibleprotein). Possible functions of the {small tilde}20 kDa proteinare discussed.
1Permanent address: Department of Biochemistry, University oflund, PO Box 124, S-221 00 Lund, Sweden 相似文献
25.
Prior eccentric contractions impair maximal insulin action on muscle glucose uptake in the conscious rat 总被引:1,自引:0,他引:1
Asp Sven; Watkinson Allan; Oakes Nicholas D.; Kraegen Edward W. 《Journal of applied physiology》1997,82(4):1327-1332
Asp, Sven, Allan Watkinson, Nicholas D. Oakes, and Edward W. Kraegen. Prior eccentric contractions impair maximal insulin action on muscle glucose uptake in the conscious rat.J. Appl. Physiol. 82(4):1327-1332, 1997.Our aim was to examine the effect of prioreccentric contractions on insulin action locally in muscle in theintact conscious rat. Anesthetized rats performed one-leg eccentriccontractions through the use of calf muscle electrical stimulationfollowed by stretch of the active muscles. Two days later, basal andeuglycemic clamp studies were conducted with the rats in the awakefasted state. Muscle glucose metabolism was estimated from2-[14C(U)]deoxy-D-glucoseandD-[3-3H]glucose administration, and comparisons were made between the eccentrically stimulated and nonstimulated (control) calfmuscles. At midphysiological insulin levels, effects ofprior eccentric exercise on muscle glucose uptake were notstatistically significant. Maximal insulin stimulation revealed reducedincremental glucose uptake above basal(P < 0.05 in the red gastrocnemius;P < 0.1 in the white gastrocnemiusand soleus) and impaired net glycogen synthesis in all eccentricallystimulated muscles (P < 0.05). Weconclude that prior eccentric contractions impair maximal insulin action (responsiveness) on local muscle glucose uptake and glycogen synthesis in the conscious rat. 相似文献
26.
Sven Erik Godtfredsen Anne Munk Rasmussen Martin Ottesen Peer Rafn Nicolai Peitersen 《Applied microbiology and biotechnology》1984,20(1):23-28
Summary Acetolactate decarboxylase activity has been detected among three genera, nine species and 263 strains of lactic acid bacteria tested in the course of a screening for acetolactate decarboxylases amenable for use in brewing as maturation aid. Streptococcus diacetylactis strain FD-64-D was found to generate a decarboxylase exhibiting a satisfactory activity and an excellent stability at the pH prevailing in beer and wort. This decarboxylase could not be solubilized but enzymatically active, freeze-dried cells were effective for satisfactory flavour maturation of beer although difficulties were encountered during attempts to remove the applied cell material by filtration of the beer. Lactobacillus casei DSM 2547 was likewise found to produce a decarboxylase exhibiting a satisfactory activity and stability at the low pH of beer and which, in addition, was readily solubilized. A method has been developed for pilot scale production of preparations of this decarboxylase suitable for use in brewing.Abbreviations DSM
Deutsche Sammlung von Microorganismen
- EDTA
Ethylene diaminetetra-acetic acid 相似文献
27.
Christer Jansson Örjan Hansson Hans-Erik Åkerlund Lars-Erik Andreasson 《Biochemical and biophysical research communications》1984,124(1):269-276
EPR measurements on inside-out thylakoids revealed that salt-washing, known to inhibit oxygen evolution and release a 23 and a 16 kDa protein, induced a Signal IIf and decreased the EPR signal from state S2. Readdition of the released 23 kDa protein restored the oxygen evolution and decreased the Signal IIf, but did not relieve the decrease in the state S2 signal. It is suggested that salt-washing inhibits the electron transfer from the oxygen-evolving site to Z, the physiological donor to P680. In inhibited photosystem II units lacking Signal IIf, Z+ is rapidly reduced, possibly by a modified S-cycle unable to evolve oxygen. 相似文献
28.
Transformation of leukotriene A4 methyl ester to leukotriene C4 monomethyl ester by cytosolic rat glutathione transferases 总被引:6,自引:0,他引:6
Bengt Mannervik Helgi Jensson Per Ålin Lars Örning Sven Hammarström 《FEBS letters》1984,175(2):289-293
Six major basic cytosolic glutathione transferases from rat liver catalyzed the conversion of leukotriene A4 methyl ester to the corresponding leukotriene C4 monomethyl ester. Glutathione transferase 4-4, the most active among these enzymes, had a Vmax of 615 nmol X min-1 X mg protein-1 at 30 degrees C in the presence of 5 mM glutathione. It was followed in efficiency by transferase 3-4 which had a Vmax of 160 nmol X min-1 X mg-1 under the same conditions. Transferases 1-1, 1-2, 2-2 and 3-3 had at least 30 times lower Vmax values than transferase 4-4. 相似文献
29.
David A. Clark Giichi Goto Anthony Marfat E.J. Corey Sven Hammarström Bengt Samuelsson 《Biochemical and biophysical research communications》1980,94(4):1133-1139
A slow reacting substance, produced by murine mastocytoma cells, has been shown to have the structure 5(S)-hydroxy-6(R)--glutathionyl-7,9,11- leukotriene C, previously referred to as leukotriene C-2) by ultraviolet spectroscopy, amino acid analyses, lipoxygenase conversion and comparisions with a synthetic compound of known structure and stereochemistry. 相似文献
30.
Michael K. Bach John R. Brashler Sven Hammarström Bengt Samuelsson 《Biochemical and biophysical research communications》1980,93(4):1121-1126
Slow reacting substance (SRS), produced by rat peritoneal mononuclear cells after stimulation with ionophore A23187, consists of two main components (Bach, M.K. et al. (1979) J. Immunol. 122, 160–165). One of these components was recently identified as leukotriene C-1. The other component has now been identified as leukotriene D. 相似文献