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- Yoshida, M., et al.
(författare)
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Production and characterization of recombinant Phanerochaete chrysosporium cellobiose dehydrogenase in the methylotrophic yeast Pichia pastoris
- 2001
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Ingår i: Bioscience, biotechnology and biochemistry. - : Oxford University Press (OUP). - 0916-8451 .- 1347-6947. ; 65:9, s. 2050-2057
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Tidskriftsartikel (refereegranskat)abstract
- The hemoflavoenzyme cellobiose dehydrogenase (CDH) from the white-rot fungus Phanerochaete chrysosporium has been heterologously expressed in the methylotrophic yeast Pichia pastoris. After 4 days of cultivation in the induction medium, the expression level reached 1800 U/L (79 mg/L) of CDH activity, which is considerably higher than that obtained previously for wild-type CDH (wtCDH) and recombinant CDH (rCDH) produced by P. chrysosporium. Analysis with SDS-PAGE and Coomassie Brilliant Blue (CBB) staining revealed a major protein band with an approximate molecular mass of 100 kDa, which was identified as rCDH by Western blotting. The absorption spectrum of rCDH shows that the protein contains one flavin and one heme cofactor per protein molecule, as does wtCDH. The kinetic parameters for rCDH using cellobiose, ubiquinone, and cytochrome c, as well as the cellulose-binding properties of rCDH were nearly identical to those of wtCDH. From these results, we conclude that the rCDH produced by Pichia pastoris retains the catalytic and cellulose-binding properties of the wild-type enzyme, and that the Pichia expression system is well suited for high-level production of rCDH.
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- Baldini, L., et al.
(författare)
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Catalog of Long-term Transient Sources in the First 10 yr of Fermi-LAT Data
- 2021
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Ingår i: Astrophysical Journal Supplement Series. - : American Astronomical Society. - 0067-0049 .- 1538-4365. ; 256:1
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Tidskriftsartikel (refereegranskat)abstract
- We present the first Fermi Large Area Telescope (LAT) catalog of long-term gamma-ray transient sources (1FLT). This comprises sources that were detected on monthly time intervals during the first decade of Fermi-LAT operations. The monthly timescale allows us to identify transient and variable sources that were not yet reported in other Fermi-LAT catalogs. The monthly data sets were analyzed using a wavelet-based source detection algorithm that provided the candidate new transient sources. The search was limited to the extragalactic regions of the sky to avoid the dominance of the Galactic diffuse emission at low Galactic latitudes. The transient candidates were then analyzed using the standard Fermi-LAT maximum likelihood analysis method. All sources detected with a statistical significance above 4 sigma in at least one monthly bin were listed in the final catalog. The 1FLT catalog contains 142 transient gamma-ray sources that are not included in the 4FGL-DR2 catalog. Many of these sources (102) have been confidently associated with active galactic nuclei (AGNs): 24 are associated with flat-spectrum radio quasars, 1 with a BL Lac object, 70 with blazars of uncertain type, 3 with radio galaxies, 1 with a compact steep-spectrum radio source, 1 with a steep-spectrum radio quasar, and 2 with AGNs of other types. The remaining 40 sources have no candidate counterparts at other wavelengths. The median gamma-ray spectral index of the 1FLT-AGN sources is softer than that reported in the latest Fermi-LAT AGN general catalog. This result is consistent with the hypothesis that detection of the softest gamma-ray emitters is less efficient when the data are integrated over year-long intervals.
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- Casanova-Acebes, M, et al.
(författare)
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Neutrophils instruct homeostatic and pathological states in naive tissues
- 2018
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Ingår i: The Journal of experimental medicine. - : Rockefeller University Press. - 1540-9538 .- 0022-1007. ; 215:11, s. 2778-2795
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Tidskriftsartikel (refereegranskat)abstract
- Immune protection relies on the capacity of neutrophils to infiltrate challenged tissues. Naive tissues, in contrast, are believed to remain free of these cells and protected from their toxic cargo. Here, we show that neutrophils are endowed with the capacity to infiltrate multiple tissues in the steady-state, a process that follows tissue-specific dynamics. By focusing in two particular tissues, the intestine and the lungs, we find that neutrophils infiltrating the intestine are engulfed by resident macrophages, resulting in repression of Il23 transcription, reduced G-CSF in plasma, and reinforced activity of distant bone marrow niches. In contrast, diurnal accumulation of neutrophils within the pulmonary vasculature influenced circadian transcription in the lungs. Neutrophil-influenced transcripts in this organ were associated with carcinogenesis and migration. Consistently, we found that neutrophils dictated the diurnal patterns of lung invasion by melanoma cells. Homeostatic infiltration of tissues unveils a facet of neutrophil biology that supports organ function, but can also instigate pathological states.
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- Forkel, Marianne, et al.
(författare)
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Composition and functionality of the intrahepatic innate lymphoid cell-compartment in human nonfibrotic and fibrotic livers
- 2017
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Ingår i: European Journal of Immunology. - : Wiley. - 0014-2980 .- 1521-4141. ; 47:8, s. 1280-1294
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Tidskriftsartikel (refereegranskat)abstract
- Human innate lymphoid cells have been described to exist in different organs, with functional deregulation of these cells contributing to several disease states. Here, we performed the first detailed characterization of the phenotype, tissue-residency properties, and functionality of ILC1s, ILC2s, and ILC3s in the human adult and fetal liver. In addition, we investigated changes in the ILC compartment in liver fibrosis. A unique composition of tissue-resident ILCs was observed in nonfibrotic livers as compared with that in mucosal tissues, with NKp44− ILC3s accounting for the majority of total intrahepatic ILCs. The frequency of ILC2s, representing a small fraction of ILCs in nonfibrotic livers, increased in liver fibrosis and correlated directly with the severity of the disease. Notably, intrahepatic ILC2s secreted the profibrotic cytokine IL-13 when exposed to IL-33 and thymic stromal lymphopoetin (TSLP); these cytokines were produced by hepatocytes, hepatic stellate cells (HSCs), and Kupffer cells in response to TLR-3 stimulation. In summary, the present results provide the first detailed characterization of intrahepatic ILCs in human adult and fetal liver. The results indicate a role for ILC2s in human liver fibrosis, implying that targeting ILC2s might be a novel therapeutic strategy for its treatment.
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