The proximity-based protein interactome and regulatory logics of the transcription factor p65 NF-κB/RELA.

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 and biotin tagging, we identify >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. A comparison with the interactomes of two point mutants of p65 reveals that the interactions primarily require intact dimerization rather than DNA-binding properties.

Metabolic labeling and LC-MS/MS-based identification of interleukin-1α-induced secreted proteomes from epithelial cells in the presence or absence of serum.

The unbiased identification of cytokine-induced, secreted proteins from cells cultured in serum-containing medium is challenging. Here, we describe an experimental and bioinformatics workflow to label interleukin-1α-regulated proteins in living cells with the methionine analogue L-homopropargylglycine. We detail their purification and identification by means of CLICK-chemistry-based biotinylation followed by nanoHPLC-MS/MS.

Multi-level inhibition of coronavirus replication by chemical ER stress.

Coronaviruses (CoVs) are important human pathogens for which no specific treatment is available. Here, we provide evidence that pharmacological reprogramming of ER stress pathways can be exploited to suppress CoV replication. The ER stress inducer thapsigargin efficiently inhibits coronavirus (HCoV-229E, MERS-CoV, SARS-CoV-2) replication in different cell types including primary differentiated human bronchial epithelial cells, (partially) reverses the virus-induced translational shut-down, improves viability of infected cells and counteracts the CoV-mediated downregulation of IRE1α and the ER chaperone BiP.

Distinct IL-1α-responsive enhancers promote acute and coordinated changes in chromatin topology in a hierarchical manner.

How cytokine-driven changes in chromatin topology are converted into gene regulatory circuits during inflammation still remains unclear. Here, we show that interleukin (IL)-1α induces acute and widespread changes in chromatin accessibility via the TAK1 kinase and NF-κB at regions that are highly enriched for inflammatory disease-relevant SNPs. Two enhancers in the extended chemokine locus on human chromosome 4 regulate the IL-1α-inducible IL8 and CXCL1-3 genes.

K63-Ubiquitylation and TRAF6 Pathways Regulate Mammalian P-Body Formation and mRNA Decapping.

Signals and posttranslational modifications regulating the decapping step in mRNA degradation pathways are poorly defined. In this study we reveal the importance of K63-linked ubiquitylation for the assembly of decapping factors, P-body formation, and constitutive decay of instable mRNAs encoding mediators of inflammation by various experimental approaches. K63-branched ubiquitin chains also regulate IL-1-inducible phosphorylation of the P-body component DCP1a.

Overlap of female reproductive cycles explains shortened interclutch interval in a lizard with invariant clutch size (Squamata: Gekkonidae: Paroedura picta).

Variable clutch size is unambiguously an ancestral state in reptiles. Only several lizard lineages have evolved so-called invariant clutch size, where all females lay just one or two eggs per clutch. This mode of reproduction is characteristic for geckos.

Identification and functional characterization of novel phosphorylation sites in TAK1-binding protein (TAB) 1.

TAB1 was defined as a regulatory subunit of the protein kinase TAK1, which functions upstream in the pathways activated by interleukin (IL)-1, tumor necrosis factor (TNF), toll-like receptors (TLRs) and stressors. However, TAB1 also functions in the p38 MAPK pathway downstream of TAK1. We identified amino acids (aa) 452/453 and 456/457 of TAB1 as novel sites phosphorylated by TAK1 as well as by p38 MAPK in intact cells as well as in vitro.

The pig as an experimental model for colonic healing study of leakage and ischemia in colonic anastomosis.

Background: The object of this experimental study was to develop a model of anastomotic failure due to primary leakage and ischemia in porcine large bowel anastomosis.

Methods: End-to-end anastomosis was constructed at descending colon in 12 pigs. The pigs were randomly divided into three groups.

Small intestinal submucosa for reinforcement of colonic anastomosis.

Background: Different materials have been evaluated for anastomotic reinforcement to prevent gastrointestinal anastomotic leakage. In this experimental study, small intestinal submucosa (SIS) was tested as a sealing for stapled colonic anastomosis in a porcine model. The aims of this study were to determine the macroscopic and microscopic outcomes and to evaluate the safety and feasibility of applying SIS for anastomotic sealing.

Small intestinal submucosa as a bioscaffold for tissue regeneration in defects of the colonic wall.

Background: Small intestinal submucosa (SIS) has proved considerable regenerative capacity for repair of bowel wall defects at different locations. This study assesses the effectiveness of SIS in the repair of defects at a gastrointestinal location with strong bacterial contamination.

Methods: Fourteen domestic pigs had a 4.

Thymidylate synthase and dihydropyrimidine dehydrogenase mRNA expression levels: predictors for survival in colorectal cancer patients receiving adjuvant 5-fluorouracil.

Purpose: Despite adjuvant 5-fluorouracil (5-FU) therapy, approximately 30% of patients with International Union against Cancer stage II and III colorectal cancer develop recurrence. In this study, we determined the prognostic value of thymidylate synthase (TS) and dihydropyrimidine dehydrogenase (DPD) expression in colorectal cancer patients treated with adjuvant 5-FU.

Experimental Design: A real-time reverse transcription-PCR technique for quantitation of relative gene expression from paraffin-embedded specimen was established first.

Topical application of vitamin A reverses metaplasia of rat vaginal epithelium: a rapid and efficient approach to improve mucosal barrier function.

Retinyl palmitate and its metabolites retinol and retinoic acid control growth and epithelial differentiation. Systemic or local vitamin A deficiency induced by malnutrition, continuous chemical irritation or locally induced by inflammation causes squamous metaplastic changes in the epithelium of mucous membranes. We demonstrate that in an animal model topically applied retinyl palmitate can be taken up by the mucosal cells independently from the systemic supply.

Intermittent activity-induced hemobilia caused by liver hemangioma.

Background: Intestinal bleeding of unknown origin can lead to a difficult workup. Abdominal colic, melena/hematemesis, and jaundice represent the pathognomonic triad for hemobilia, but clinical presentation and etiology of this entity are varying. Seldom all of these symptoms are present, and rarely does hemobilia cause melena or hematemesis.

Vascular injuries within the hepatoduodenal ligament: recognition by laparoscopic color Doppler ultrasound.

Background: Lesions of vascular structures are rare but serious complications of laparoscopic cholecystectomy. The purpose of this blind randomized animal study was to investigate the possibility of detecting different vascular lesions within the hepatoduodenal ligament using laparoscopic color Doppler ultrasound (LCDU).

Methods: Twenty-four lesions of the hepatic artery and portal vein were created laparoscopically in six farmer pigs using titanium clips.

Laparoscopic ultrasonography reliably visualizes bile-duct injuries - a blind randomized porcine study.

Introduction: Despite use of intraoperative cholangiography (IOC), only one-third or less of bile duct injuries that occur during laparoscopic cholecystectomy are detected at the primary operation. Therefore, there is a need to investigate other intraoperative diagnostic methods, especially laparoscopic ultrasound, for detecting bile-duct injuries.

Materials And Methods: Detection of different bile-duct injuries by laparoscopic intraoperative ultrasonography was evaluated in a blind, randomized animal study.

Expression patterns of complex glycoconjugates and endogenous lectins during fetal development of the viscerocranium.

Experimental evidence suggests that carbohydrates and their corresponding receptors (endogenous lectins) decode biological information. Therefore, the expression of complex oligosaccharides--the potential ligand part of this recognition system--during chondrogenesis and osteogenesis was determined in the viscerocranium of fetal rats by mapping the staining patterns of exogenous lectins. Results were compared with the expression of bone- and/or cartilage-specific core proteins and the binding profiles of neoglycoconjugates.

[Online infection recording within the scope of total quality management].

The continuous and exact recording of infections is a condition sine qua non for total quality management. Therefore, at the Diakoniekrankenhaus Rotenburg (Wümme) a program which offers the possibility of online recording of data was integrated into the hospital intranet. The recording is done with network clients.

Impaired responses to sweet taste in vitamin A-deficient rats.

In brief-exposure, two-choice preference tests (sucrose solution v water), vitamin A-deficient (VAD) rats exhibited a decreased preference for sucrose relative to control rats. There was no difference in total fluid intake from both choices between the two groups, nor was any significant difference found in circumvallate taste papilla keratin size. It is concluded that the impaired preference for sucrose in VAD rats is due to a specific impairment in taste sensation rather than general malaise.

Prospective randomized comparison of laparoscopic ultrasonography using a flexible-tip ultrasound probe and intraoperative dynamic cholangiography during laparoscopic cholecystectomy.

Background: We performed a prospective randomized comparison of laparoscopic intraoperative ultrasonography (LIOU) and dynamic intraoperative cholangiography (IOC) during laparoscopic cholecystectomy (LC).

Methods: LIOU and IOC were attempted in 518 consecutive patients scheduled for laparoscopic cholecystectomy. The order in which the diagnostic procedures were performed was randomly assigned.

Interaction of vitamins E and K: effect of high dietary vitamin E on phylloquinone activity in chicks.

To determine the influence of vitamin E on phylloquinone activity, one day-old chicks were raised on a masch diet supplemented with different amounts of vitamin E for 31 days. In chicks fed a diet high in vitamin E (4000 mg allrac-alpha-tocopheryl acetate/kg) but adequate in vitamin K (0.14 mg phylloquinone/kg) a threefold increase in prothrombin time and an increase in mortality rate (five out of twelve animals died from increased bleeding tendency) was observed.

Stabilization of rat heart mitochondria by alpha-tocopherol in rats.

alpha-Tocopherol (alpha-T) acts by different mechanisms: scavenging free oxygen radicals (FOR) and stabilizing membranes; both may contribute to the prevention of the pathogenesis of diseases like cardiovascular events. The present work shows that alpha-T breaks the vicious cycle generated by damaged mitochondria. The latter stimulate the immune system increasing FOR formation that leads to more damaged mitochondria.