Upon phagocytosis, Legionella pneumophila translocates numerous effector proteins into host cells to perturb cellular metabolism and immunity, ultimately establishing intracellular survival and ...growth. VipD of L. pneumophila belongs to a family of bacterial effectors that contain the N-terminal lipase domain and the C-terminal domain with an unknown function. We report the crystal structure of VipD and show that its C-terminal domain robustly interferes with endosomal trafficking through tight and selective interactions with Rab5 and Rab22. This domain, which is not significantly similar to any known protein structure, potently interacts with the GTP-bound active form of the two Rabs by recognizing a hydrophobic triad conserved in Rabs. These interactions prevent Rab5 and Rab22 from binding to downstream effectors Rabaptin-5, Rabenosyn-5 and EEA1, consequently blocking endosomal trafficking and subsequent lysosomal degradation of endocytic materials in macrophage cells. Together, this work reveals endosomal trafficking as a target of L. pneumophila and delineates the underlying molecular mechanism.
This paper proposes a method to improve the performance of a distribution system by optimizing volt–var function of a smart inverter to alleviate the voltage deviation problem due to distributed ...generation connection. In order to minimize voltage deviation and line losses which represent the performance of a distribution system, this paper proposes an algorithm that optimally sets the parameters of the volt–var function. In the process of optimizing the parameters of the volt–var function, the algorithm proposed in this paper considers minimizing the contribution of the reactive power in order not to affect the output of the distributed generation. In order to apply to the field, the distribution system in South Korea considering the configuration and operation regulation was selected as a test model for algorithm verification. As a result, the system performance was successfully improved by optimally setting the volt–var function of the smart inverter which is an effective way to solve the over-voltage problem caused by distributed generation connection. This paper verified the proposed method through OpenDSS, a quasi-static time-series simulation, for the test model considering the characteristics of the distribution system in South Korea.
Most industrial technologies are automated by sensitive electric parts such as programmable logic controllers(PLCs) and magnetic contactors which are required high power quality. The voltage sags ...occur in the non-faulted power distribution feeders during the fault duration. Therefore, voltage causes economic losses in the production process of various companies. To solve the fault current and the voltage sags, the superconducting fault current limiter (SFCL) is in the spotlight. The SFCL where was located on starting point of the feeder improves the reliability of the facility by reducing the fault current. In addition, it compensates the voltage sags on the bus-bar. In this paper, evaluating the voltage sags based on fault location in the power distribution system with the SFCL through PSCAD/EMTDC simulation. Next, the expected the voltage sags frequency is predicted by the reliability analysis technique of the power distribution system. The expected voltage sags frequency of the power distribution feeder is compared with before and after the application of the SFCL.
FcγRIIIa, which is predominantly expressed on the surface of natural killer cells, plays a key role in antibody-dependent cell-mediated cytotoxicity (ADCC), a major effector function of therapeutic ...IgG antibodies that results in the death of aberrant cells. Despite the potential uses of aglycosylated IgG antibodies, which can be easily produced in bacteria and do not have complicated glycan heterogeneity issues, they show negligible binding to FcγRIIIa and abolish the activation of immune leukocytes for tumor cell clearance, in sharp contrast to most glycosylated IgG antibodies used in the clinical setting. For directed evolution of aglycosylated Fc variants that bind to FcγRIIIa and, in turn, exert potent ADCC effector function, we randomized the aglycosylated Fc region of full-length IgG expressed on the inner membrane of Escherichia coli. Multiple rounds of high-throughput screening using flow cytometry facilitated the isolation of aglycosylated IgG Fc variants that exhibited higher binding affinity to FcγRIIIa-158V and FcγRIIIa-158F compared with clinical-grade trastuzumab (Herceptin®). The resulting aglycosylated trastuzumab IgG antibody Fc variants could elicit strong peripheral blood mononuclear cell-mediated ADCC without glycosylation in the Fc region.
The sweet-tasting protein brazzein offers considerable potential as a functional sweetener with antioxidant, anti-inflammatory, and anti-allergic properties. Here, we optimized a chemically defined ...medium to produce secretory recombinant brazzein in
Kluyveromyces lactis
, with applications in mass production. Compositions of defined media were investigated for two phases of fermentation: the first phase for cell growth, and the second for maximum brazzein secretory production. Secretory brazzein expressed in the optimized defined medium exhibited higher purity than in the complex medium; purification was by ultrafiltration using a molecular weight cutoff, yielding approximately 107 mg L
−1
. Moreover, the total media cost in this defined medium system was approximately 11% of that in the optimized complex medium to generate equal amounts of brazzein. Therefore, the
K. lactis
expression system is useful for mass-producing recombinant brazzein with high purity and yield at low production cost and indicates a promising potential for applications in the food industry.
Three-dimensional imaging is still not in the full commercial stage, but its application is widening due to its capability of creating real-world-like environments. This capability is especially ...important in realizing reality communication and telepresence operations in medical and unreachable places. For these applications, technologies for interacting with objects in the 3-D image should also be developed. The widespread use of 3-D images is expected in near future, but before that, problems of minimizing physical stresses, especially eye fatigue, should be solved for 3-D imaging. Currently, expected solutions to the problems are electroholography and super multiview methods. These two methods work at different principles, but they both can provide the continuous parallax as in the real-world scenes/objects. Electroholography can also provide focusable depth information to the viewers, though its viewing zone angle is limited. For the super multiview, it is not apparent that it can provide the focusable depth information, but it can provide more comfortable viewing condition than the holography because of its wider viewing zone angle.
Total polyphenol and total flavonoid assays were performed to characterize the relationships between the color of Peucedanum japonicum (PJ) seed coat and stem and the content of phytochemical ...compounds. The samples were divided into two groups based on their stem and seed coat color, with each group containing 23 samples. The stem color group was subdivided into green, light red, and red, whereas the seed coat color group was divided into light brown, brown, and dark brown. In the stem color group, the light red stems exhibited the highest content of phytochemical compounds, with levels over 10% higher than those of the stems of the other colors. Moreover, among the top ten samples with the highest total polyphenol content, eight samples were light red, and the light red group also exhibited the highest total flavonoid content among the examined color groups. In terms of the seed coat color, the plants grown from dark brown seeds exhibited the highest contents of both total polyphenols and total flavonoids. In conclusion, PJ plants with dark brown seeds and light red stems contained the highest levels of phytochemical compounds. Collectively, our findings provide a valuable basis for future seed selection of PJ for pharmaceutical purposes.
All gammaherpesviruses express homologues of antiapoptotic B-cell lymphoma-2 (BCL-2) to counter the clearance of infected cells by host antiviral defense machineries. To gain insights into the action ...mechanisms of these viral BCL-2 proteins, we carried out structural and biochemical analyses on the interactions of M11, a viral BCL-2 of murine gamma-herpesvirus 68, with a fragment of proautophagic Beclin1 and BCL-2 homology 3 (BH3) domain-containing peptides derived from an array of proapoptotic BCL-2 family proteins. Mainly through hydrophobic interactions, M11 bound the BH3-like domain of Beclin1 with a dissociation constant of 40 nanomole, a markedly tighter affinity compared to the 1.7 micromolar binding affinity between cellular BCL-2 and Beclin1. Consistently, M11 inhibited autophagy more efficiently than BCL-2 in NIH3T3 cells. M11 also interacted tightly with a BH3 domain peptide of BAK and those of the upstream BH3-only proteins BIM, BID, BMF, PUMA, and Noxa, but weakly with that of BAX. These results collectively suggest that M11 potently inhibits Beclin1 in addition to broadly neutralizing the proapoptotic BCL-2 family in a similar but distinctive way from cellular BCL-2, and that the Beclin1-mediated autophagy may be a main target of the virus.
Orphan nuclear receptor small heterodimer partner (SHP) plays a key role in transcriptional repression of gluconeogenic enzyme gene expression. Here, we show that SHP inhibited protein kinase ...A-mediated transcriptional activity of cAMP-response element-binding protein (CREB), a major regulator of glucose metabolism, to modulate hepatic gluconeogenic gene expression. Deletion analysis of phosphoenolpyruvate carboxykinase (PEPCK) promoter demonstrated that SHP inhibited forskolin-mediated induction of PEPCK gene transcription via inhibition of CREB transcriptional activity. In vivo imaging demonstrated that SHP inhibited CREB-regulated transcription coactivator 2 (CRTC2)-mediated cAMP-response element-driven promoter activity. Furthermore, overexpression of SHP using adenovirus SHP decreased CRTC2-dependent elevations in blood glucose levels and PEPCK or glucose-6-phosphatase (G6Pase) expression in mice. SHP and CREB physically interacted and were co-localized in vivo. Importantly, SHP inhibited both wild type CRTC2 and S171A (constitutively active form of CRTC2) coactivator activity and disrupted CRTC2 recruitment on the PEPCK gene promoter. In addition, metformin or overexpression of a constitutively active form of AMPK (Ad-CA-AMPK) inhibited S171A-mediated PEPCK and G6Pase gene expression, and hepatic glucose production and knockdown of SHP partially relieved the metformin- and Ad-CA-AMPK-mediated repression of hepatic gluconeogenic enzyme gene expression in primary rat hepatocytes. In conclusion, our results suggest that a delayed effect of metformin-mediated induction of SHP gene expression inhibits CREB-dependent hepatic gluconeogenesis.
BACKGROUND/AIMSThe risk of herpes zoster (HZ) is increased in patients with autoimmune diseases (AID), probably due to immunosuppressive therapy. METHODSThis prospective cross-sectional study ...investigated varicella zoster virus (VZV)-specific immunity in relation to subclinical VZV reactivation in 48 AID patients and 48 healthy controls (HCs). We assessed humoral immunity (serum VZV immunoglobulin g IgG, IgA, and IgM) and cell-mediated immunity (interferon-γ IFNγ-releasing assay) to VZV as well as salivary VZV DNA status. Subclinical VZV reactivation was confirmed by detecting VZV DNA in saliva or VZV IgM in serum in the absence of typical HZ symptoms. RESULTSMedian IgA levels were higher in the AID group than in the HC group, while VZV IgG and IgM levels were comparable between the groups. AID patients showed fewer IFNγ spot-forming cells (SFCs) upon VZV stimulation than HCs (58.2 vs. 122.0 SFCs/106 peripheral blood mononuclear cells PBMCs, p < 0.0001). Subclinical VZV reactivation was more frequent in AID patients than in HCs (12.5% vs. 0%, p = 0.01). AID patients with VZV reactivation received prednisolone more frequently and at a higher dose than AID patients without reactivation. VZV-specific IFNγ SFCs were significantly lower in patients with VZV reactivation among AID patients (26.3 vs. 62.6 SFCs/106 PBMCs, p < 0.0001). CONCLUSIONResults suggest that poor cellular response against VZV might cause clinical and subclinical reactivation of VZV in AID patients.
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