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The NF-κB signaling system plays an important regulatory role in the control of many biological processes. The activities of NF-κB signaling networks and the expression of their target genes are frequently elevated in pathophysiological situations including inflammation, infection, and cancer. In these conditions, the outcome of NF-κB activity can vary according to (i) differential activation states, (ii) the pattern of genomic recruitment of the NF-κB subunits, and (iii) cellular heterogeneity. Additionally, the cytosolic NF-κB activation steps leading to the liberation of DNA-binding dimers need to be distinguished from the less understood nuclear pathways that are ultimately responsible for NF-κB target gene specificity. This raises the need to more precisely determine the NF-κB activation status not only for the purpose of basic research, but also in (future) clinical applications. Here we review a compendium of different methods that have been developed to assess the NF-κB activation status in vitro and in vivo. We also discuss recent advances that allow the assessment of several NF-κB features simultaneously at the single cell level.

The protein interactome of p65/RELA, the most active subunit of the transcription factor (TF) NF-kappaB, has not been previously determined in living cells. Using p65-miniTurbo fusion proteins, we identified by biotin tagging > 350 RELA interactors from untreated and IL-1alpha-stimulated cells, including many TFs (47 % of all interactors) and > 50 epigenetic regulators belonging to different classes of chromatin remodeling complexes. According to point mutants of p65, the interactions primarily require intact dimerization rather than DNA binding properties. A targeted RNAi screen for 38 interactors and subsequent functional transcriptome and bioinformatics studies identified gene regulatory (sub)networks, each controlled by RELA in combination with one of the TFs ZBTB5, GLIS2, TFE3/TFEB or S100A8/A9. The remarkably large, dynamic and versatile high resolution interactome of RELA and its gene-regulatory logics provides a rich resource and a new framework for explaining how RELA cooperativity determines gene expression patterns.Competing Interest StatementThe authors have declared no competing interest.

The protein interactome of p65 / RELA, the most active subunit of the transcription factor (TF) NF-κB, has not been previously determined in living cells. Using p65-miniTurbo fusion proteins, we identified by biotin tagging > 350 RELA interactors from untreated and IL-1α-stimulated cells, including many TFs (47 % of all interactors) and > 50 epigenetic regulators belonging to different classes of chromatin remodeling complexes. According to point mutants of p65, the interactions primarily require intact dimerization rather than DNA binding properties. A targeted RNAi screen for 38 interactors and subsequent functional transcriptome and bioinformatics studies identified gene regulatory (sub)networks, each controlled by RELA in combination with one of the TFs ZBTB5, GLIS2, TFE3 / TFEB or S100A8 / A9. The remarkably large, dynamic and versatile high resolution interactome of RELA and its gene-regulatory logics provides a rich resource and a new framework for explaining how RELA cooperativity determines gene expression patterns. Identification of > 350 largely dimerization-dependent interactors of p65 / RELA by miniTurboID The interactome is dominated by transcription factors and epigenetic regulator complexes Functional validation of 38 high confidence interactors by targeted siRNA screen Identification of genetic networks regulated by RELA and six of its interactors in the IL-1α response

We provide recommendations for sampling and identification of introduced larval parasitoids of spotted-wing drosophila, Drosophila suzukii (Matsumura) (Diptera: Drosophilidae). These parasitoids are either under consideration for importation (aka classical) biological control introductions, or their adventive (presumed to have been accidentally introduced) populations have recently been discovered in North America and Europe. Within the context of the ecology of D. suzukii and its parasitoids, we discuss advantages and disadvantages of estimating larval parasitism levels using different methods, including naturally collected fruit samples and sentinel baits. For most situations, we recommend repeated sampling of naturally occurring fruit rather than using sentinel baits to monitor seasonal dynamics of host plant–Drosophila–parasitoid associations. We describe how to separate Drosophilidae puparia from host fruit material in order to accurately estimate parasitism levels and establish host–parasitoid associations. We provide instructions for identification of emerging parasitoids and include a key to the common families of parasitoids of D. suzukii. We anticipate that the guidelines for methodology and interpretation of results that we provide here will form the basis for a large, multi-research team sampling effort in the coming years to characterize the biological control and nontarget impacts of accidentally and intentionally introduced larval parasitoids of D. suzukii in several regions of the world.

Open access funding provided by Fondazione Edmund Mach - Istituto Agrario di San Michele all'Adige within the CRUI-CARE Agreement. This research was supported by multiple funding sources. In the USA, funding were provided by USDA Animal and Plant Health Inspection Service (APHIS) Farm Bill Award No. 60–8010-4–001, USDA base funds: CRIS 2072–22000-044-00D, USDA Agricultural Research Service (ARS) Areawide Program (National Program Leader: S. Young), USDA Specialty Crop Research Initiative (SCRI) Award No. 2020–51181-32140, USDA Crop Protection and Pest Management (CPPM) Award No. 2021–70006-35312, USDA National Institute of Food and Agriculture (NIFA) Awards No. 2021–70006-35312 and 2020–51181-32140, USDA Specialty Crop Block Grant Program (SCBGP), USDA Northwest Center for Small Fruit Research, Oregon Blueberry Commission, Oregon Blackberry Commission, New Jersey Blueberry and Cranberry Research Council, Michigan Blueberry Commission and the “GREEEN—Generating Research and Extension to meet Economic and Environmental Needs” project. In Canada, funding was provided by the Dekaban Scholars program, the Natural Sciences and Engineering Research Council of Canada (NSERC) and the Agriculture and Agri-Food Canada: Projects J-003401 and J-002839. In France, field surveys by INRAE ISA were part of the Ecophyto SUZoCARPO project, funded by the French Ministry of Agriculture and Food. In Switzerland, field surveys were funded by the Swiss Federal Office for Agriculture. In Italy, funding were provided by the Samurai Wasp Action Team (SWAT) project, funded by the Autonomous Province of Trento, the project \"URCOFI—Unità Regionale Coordinamento Fitosanitario,\" funded by the Campania Region government (Grant No. CUP B29I22001290009), the project “SUccess of Specialist versus generalist parasitoid in Hampering the spread of an Invasive pest (SUSHI),” funded by the Italian Ministry of University and Research (MUR) (Grant No. 20227ZYCH3, CUP Master B53D2301184 0006).

[SARAH BARRINGER GORDON] began Yale Law School in the early 1980s with an unusual goal: to attend divinity school and law school simultaneously and earn both degrees. \"I wrestled with questions of separation of church and state, individual religious liberty- and even the definition of 'religion,\"' she explains in the preface of The Mormon Question: Polygamy and Constitutional Conflict in NineteenthCentury America (Univ. of North Carolina, Jan.).

Cheyenne was found unharmed at about 11 a.m. on Wednesday, after a greeter at a 24-hour Walmart in north Edmonton noticed the missing girl inside the store. The group spoke with the girl and asked her some basic questions, such as what her name was and what school she attended.