NOTCH1 can initiate NF-κB activation via cytosolic interactions with components of the T cell Signalosome.

T cell stimulation requires the input and integration of external signals. Signaling through the T cell receptor (TCR) is known to induce formation of the membrane-tethered CBM complex, comprising CARMA1, BCL10, and MALT1, which is required for TCR-mediated NF-κB activation. TCR signaling has been shown to activate NOTCH proteins, transmembrane receptors also implicated in NF-κB activation. However, the link between TCR-mediated NOTCH signaling and early events leading to induction of NF-κB activity remains unclear. In this report, we demonstrate a novel cytosolic function for NOTCH1 and show that it is essential to CBM complex formation. Using a model of skin allograft rejection, we show in vivo that NOTCH1 acts in the same functional pathway as PKCθ, a T cell-specific kinase important for CBM assembly and classical NF-κB activation. We further demonstrate in vitro NOTCH1 associates physically with PKCθ and CARMA1 in the cytosol. Unexpectedly, when NOTCH1 expression was abrogated using RNAi approaches, interactions between CARMA1, BCL10, and MALT1 were lost. This failure in CBM assembly reduced inhibitor of kappa B alpha phosphorylation and diminished NF-κB-DNA binding. Finally, using a luciferase gene reporter assay, we show the intracellular domain of NOTCH1 can initiate robust NF-κB activity in stimulated T cells, even when NOTCH1 is excluded from the nucleus through modifications that restrict it to the cytoplasm or hold it tethered to the membrane. Collectively, these observations provide evidence that NOTCH1 may facilitate early events during T cell activation by nucleating the CBM complex and initiating NF-κB signaling

Pharmacological inhibition of ULK1 kinase blocks mammalian target of rapamycin (mTOR)-dependent autophagy

Autophagy is a cell-protective and degradative process that recycles damaged and long-lived cellular components. Cancer cells are thought to take advantage of autophagy to help them to cope with the stress of tumorigenesis; thus targeting autophagy is an attractive therapeutic approach. However, there are currently no specific inhibitors of autophagy. ULK1, a serine/threonine protein kinase, is essential for the initial stages of autophagy, and here we report that two compounds, MRT67307 and MRT68921, potently inhibit ULK1 and ULK2 in vitro and block autophagy in cells. Using a drug-resistant ULK1 mutant, we show that the autophagy-inhibiting capacity of the compounds is specifically through ULK1. ULK1 inhibition results in accumulation of stalled early autophagosomal structures, indicating a role for ULK1 in the maturation of autophagosomes as well as initiation

Chimeric Anti-Staphylococcal Enterotoxin B Antibodies and Lovastatin Act Synergistically to Provide In Vivo Protection against Lethal Doses of SEB

Staphylococcal enterotoxin B (SEB) is one of a family of toxins secreted by Staphylococcus aureus that act as superantigens, activating a large fraction of the T-cell population and inducing production of high levels of inflammatory cytokines that can cause toxic shock syndrome (TSS) and death. Extracellular engagement of the TCR of T-cells and class II MHC of antigen presenting cells by SEB triggers the activation of many intracellular signaling processes. We engineered chimeric antibodies to block the extracellular engagement of cellular receptors by SEB and used a statin to inhibit intracellular signaling. Chimeric human-mouse antibodies directed against different neutralizing epitopes of SEB synergistically inhibited its activation of human T-cells in vitro. In the in vivo model of lethal toxic shock syndrome (TSS) in HLA-DR3 transgenic mice, two of these antibodies conferred significant partial protection when administered individually, but offered complete protection in a synergistic manner when given together. Similarly, in vivo, lovastatin alone conferred only partial protection from TSS similar to single anti-SEB antibodies. However, used in combination with one chimeric neutralizing anti-SEB antibody, lovastatin provided complete protection against lethal TSS in HLA-DR3 transgenic mice. These experiments demonstrate that in vivo protection against lethal doses of SEB can be achieved by a statin of proven clinical safety and chimeric human-mouse antibodies, agents now widely used and known to be of low immunogenicity in human hosts

Incidence, management, and outcomes of cardiovascular insufficiency in critically ill term and late preterm newborn infants

OBJECTIVE: The objective of this study was to characterize the incidence, management, and short-term outcomes of cardiovascular insufficiency (CVI) in mechanically ventilated newborns, evaluating four separate prespecified definitions. STUDY DESIGN: Multicenter, prospective cohort study of infants ≥34 weeks gestational age (GA) and on mechanical ventilation during the first 72 hours. CVI was prospectively defined as either (1) mean arterial pressure (MAP) < GA; (2) MAP < GA + signs of inadequate perfusion; (3) any therapy for CVI; or (4) inotropic therapy. Short-term outcomes included death, days on ventilation, oxygen, and to full feedings and discharge. RESULTS: Of 647 who met inclusion criteria, 419 (65%) met ≥1 definition of CVI. Of these, 98% received fluid boluses, 36% inotropes, and 17% corticosteroids. Of treated infants, 46% did not have CVI as defined by a MAP < GA ± signs of inadequate perfusion. Inotropic therapy was associated with increased mortality (11.1 vs. 1.3%; p < 0.05). CONCLUSION: More than half of the infants met at least one definition of CVI. However, almost half of the treated infants met none of the definitions. Inotropic therapy was associated with increased mortality. These findings can help guide the design of future studies of CVI in newborn

The Vehicle, Spring 1974

Table of Contents PhotoJim Painterpage 1 Six Poems of the LandRay Schmuddepage 5 At Last to Find FreedomJann Briesacherpage 7 The Last IrisMarjorie Thoelepage 9 (Untitled)Melinda E. Recordpage 10 MenJan Schroederpage 10 ImpressionsJudy Bardpage 11 ScaredAnita Surpage 11 Loved and LostJan Schroederpage 12 Dripped-Over WaxAnita Surpage 13 The Crowded RoomWilliam E. Uteschpage 14 A River in IllinoisJames Jonespage 14 Sneeze SeasonDarlene A. Moorepage 14 ChangesMark Chianakaspage 15 PhotoJim Painterpage 16 Wedding VowsJann Briesacherpage 17 PhotoJim Painterpage 18 PhotoJim Painterpage 19 PhotoJim Painterpage 20 PhotoJim Painterpage 21 PhotoJim Painterpage 22 PhotoLarry Smyserpage 23 From Outside ColoradoRay Schmuddepage 24 Dairy QueenGayle Gleichmanpage 26 With Sunstreaks in our HairNancy Broom Brownpage 33 PhotoJim Painterpage 34 Water\u27s EdgeMarjorie Thoelepage 35 My 665th Illusion of SanityGordon Glessnerpage 36 Is it my turn to do the laundry again??? Jann Briesacherpage 38 TV Teachingbobbdoddpage 39 GuidanceWendy Diane Wielandpage 40 PhotoJim Painterpage 41 RaindropsJane Ann Beerspage 42 WaitingJan Schroederpage 42 To JonJudy Bardpage 43 One Autumn Day in 1971E. Christmanpage 43 More Surely Than Picture AlbumsMarjorie Thoelepage 44 WingspanningNancy Broom Brownpage 45 ReligionMelinda E. Recordpage 45 Rosalie StevensonMark Holleypage 46 PhotoJim Painterpage 47 WhiteShirley A. Rardinpage 48 The Beginning of a Perfect DayShirley A. Rardinpage 49 PhotoMichael Chenpage 50 Rosethorn Wall of June 17bobbdoddpage 51 ManJan Schroederpage 51 HaikuJudy Bardpage 51 You know it leaves me emptyJames Osbornepage 52 For JesseJames Osbornepage 52 EndingsMark Chianakaspage 53 ConfusionGary L. Owenspage 53 PhotoMichael Chenpage 54 PoemsJann Briesacherpage 54 Journey of just oneNancy Broom Brownpage 55 Blackbirds in IllinoisJames Jonespage 56 PoemsJann Briesacherpage 56 PhotoMichael Chenpage 57 I am a poemDarlene A. Moorepage 57 A Glimpse of ParadiseJann Briesacherpage 57 PhotoJim Painterpage 58 PoemSheila Marie Foorpage 59 In my windowBarbara S. Meyerpage 59 Section 4., Draft 3bobbdoddpage 60 PhotoJim Painterpage 61 PoemJann Briesacherpage 61 PhotoGary Deanpage 62 I amWilliam E. Uteschpage 62 To a tank-car in IllinoisJames Jonespage 63 PoemJane Ann Beerspage 63 PoemsJann Briesacherpage 63 Editor\u27s Pagepage 64https://thekeep.eiu.edu/vehicle/1031/thumbnail.jp

The Vehicle, Spring 1974

Table of Contents PhotoJim Painterpage 1 Six Poems of the LandRay Schmuddepage 5 At Last to Find FreedomJann Briesacherpage 7 The Last IrisMarjorie Thoelepage 9 (Untitled)Melinda E. Recordpage 10 MenJan Schroederpage 10 ImpressionsJudy Bardpage 11 ScaredAnita Surpage 11 Loved and LostJan Schroederpage 12 Dripped-Over WaxAnita Surpage 13 The Crowded RoomWilliam E. Uteschpage 14 A River in IllinoisJames Jonespage 14 Sneeze SeasonDarlene A. Moorepage 14 ChangesMark Chianakaspage 15 PhotoJim Painterpage 16 Wedding VowsJann Briesacherpage 17 PhotoJim Painterpage 18 PhotoJim Painterpage 19 PhotoJim Painterpage 20 PhotoJim Painterpage 21 PhotoJim Painterpage 22 PhotoLarry Smyserpage 23 From Outside ColoradoRay Schmuddepage 24 Dairy QueenGayle Gleichmanpage 26 With Sunstreaks in our HairNancy Broom Brownpage 33 PhotoJim Painterpage 34 Water\u27s EdgeMarjorie Thoelepage 35 My 665th Illusion of SanityGordon Glessnerpage 36 Is it my turn to do the laundry again??? Jann Briesacherpage 38 TV Teachingbobbdoddpage 39 GuidanceWendy Diane Wielandpage 40 PhotoJim Painterpage 41 RaindropsJane Ann Beerspage 42 WaitingJan Schroederpage 42 To JonJudy Bardpage 43 One Autumn Day in 1971E. Christmanpage 43 More Surely Than Picture AlbumsMarjorie Thoelepage 44 WingspanningNancy Broom Brownpage 45 ReligionMelinda E. Recordpage 45 Rosalie StevensonMark Holleypage 46 PhotoJim Painterpage 47 WhiteShirley A. Rardinpage 48 The Beginning of a Perfect DayShirley A. Rardinpage 49 PhotoMichael Chenpage 50 Rosethorn Wall of June 17bobbdoddpage 51 ManJan Schroederpage 51 HaikuJudy Bardpage 51 You know it leaves me emptyJames Osbornepage 52 For JesseJames Osbornepage 52 EndingsMark Chianakaspage 53 ConfusionGary L. Owenspage 53 PhotoMichael Chenpage 54 PoemsJann Briesacherpage 54 Journey of just oneNancy Broom Brownpage 55 Blackbirds in IllinoisJames Jonespage 56 PoemsJann Briesacherpage 56 PhotoMichael Chenpage 57 I am a poemDarlene A. Moorepage 57 A Glimpse of ParadiseJann Briesacherpage 57 PhotoJim Painterpage 58 PoemSheila Marie Foorpage 59 In my windowBarbara S. Meyerpage 59 Section 4., Draft 3bobbdoddpage 60 PhotoJim Painterpage 61 PoemJann Briesacherpage 61 PhotoGary Deanpage 62 I amWilliam E. Uteschpage 62 To a tank-car in IllinoisJames Jonespage 63 PoemJane Ann Beerspage 63 PoemsJann Briesacherpage 63 Editor\u27s Pagepage 64https://thekeep.eiu.edu/vehicle/1031/thumbnail.jp

c-Jun N-Terminal Kinase 1 Is Required for Toll-Like Receptor 1 Gene Expression in Macrophages

P. 5027-5034The regulation of innate immune responses to pathogens occurs through the interaction of Toll-like receptors (TLRs) with pathogen-associated molecular patterns and the activation of several signaling pathways whose contribution to the overall innate immune response to pathogens is poorly understood. We demonstrate a mechanism of control of murine macrophage responses mediated by TLR1/2 heterodimers through c-Jun N-terminal kinase 1 (JNK1) activity. JNK controls tumor necrosis factor alpha production and TLR-mediated macrophage responses to Borrelia burgdorferi, the causative agent of Lyme disease, and the TLR1/TLR2-specific agonist PAM3CSK4. JNK1, but not JNK2, activity regulates the expression of the tlr1 gene in the macrophage cell line RAW264.7, as well as in primary CD11b cells. We also show that the proximal promoter region of the human tlr1 gene contains an AP-1 binding site that is subjected to regulation by the kinase and binds two complexes that involve the JNK substrates c-Jun, JunD, and ATF-2. These results demonstrate that JNK1 regulates the response to TLR1/2 ligands and suggest a positive feedback loop that may serve to increase the innate immune response to the spirocheteS

The state of the Martian climate

60°N was +2.0°C, relative to the 1981–2010 average value (Fig. 5.1). This marks a new high for the record. The average annual surface air temperature (SAT) anomaly for 2016 for land stations north of starting in 1900, and is a significant increase over the previous highest value of +1.2°C, which was observed in 2007, 2011, and 2015. Average global annual temperatures also showed record values in 2015 and 2016. Currently, the Arctic is warming at more than twice the rate of lower latitudes

Notch Signaling Regulates Mitochondrial Metabolism and NF-κB Activity in Triple-Negative Breast Cancer Cells via IKKα-Dependent Non-canonical Pathways

Triple negative breast cancer (TNBC) patients have high risk of recurrence and metastasis, and current treatment options remain limited. Cancer stem-like cells (CSCs) have been linked to cancer initiation, progression and chemotherapy resistance. Notch signaling is a key pathway regulating TNBC CSC survival. Treatment of TNBC with PI3K or mTORC1/2 inhibitors results in drug-resistant, Notch-dependent CSC. However, downstream mechanisms and potentially druggable Notch effectors in TNBC CSCs are largely unknown. We studied the role of the AKT pathway and mitochondrial metabolism downstream of Notch signaling in TNBC CSC from cell lines representative of different TNBC molecular subtypes as well as a novel patient-derived model. We demonstrate that exposure of TNBC cells to recombinant Notch ligand Jagged1 leads to rapid AKT phosphorylation in a Notch1-dependent but RBP-Jκ independent fashion. This requires mTOR and IKKα. Jagged1 also stimulates mitochondrial respiration and fermentation in an AKT- and IKK-dependent fashion. Notch1 co-localizes with mitochondria in TNBC cells. Pharmacological inhibition of Notch cleavage by gamma secretase inhibitor PF-03084014 in combination with AKT inhibitor MK-2206 or IKK-targeted NF-κB inhibitor Bay11-7082 blocks secondary mammosphere formation from sorted CD90hi or CD44+CD24low (CSCs) cells. A TNBC patient-derived model gave comparable results. Besides mitochondrial oxidative metabolism, Jagged1 also triggers nuclear, NF-κB-dependent transcription of anti-apoptotic gene cIAP-2. This requires recruitment of Notch1, IKKα and NF-κB to the cIAP-2 promoter. Our observations support a model where Jagged1 triggers IKKα-dependent, mitochondrial and nuclear Notch1 signals that stimulate AKT phosphorylation, oxidative metabolism and transcription of survival genes in PTEN wild-type TNBC cells. These data suggest that combination treatments targeting the intersection of the Notch, AKT and NF-κB pathways have potential therapeutic applications against CSCs in TNBC cases with Notch1 and wild-type PTEN expression