BRD9 regulates interferon-stimulated genes during macrophage activation via cooperation with BET protein BRD4

Macrophages induce a number of inflammatory response genes in response to stimulation with microbial ligands. In response to endotoxin Lipid A, a gene-activation cascade of primary followed by secondary-response genes is induced. Epigenetic state is an important regulator of the kinetics, specificity, and mechanism of gene activation of these two classes. In particular, SWI/SNF chromatin-remodeling complexes are required for the induction of secondary-response genes, but not primary-response genes, which generally exhibit open chromatin. Here, we show that a recently discovered variant of the SWI/SNF complex, the noncanonical BAF complex (ncBAF), regulates secondary-response genes in the interferon (IFN) response pathway. Inhibition of bromodomain-containing protein 9 (BRD9), a subunit of the ncBAF complex, with BRD9 bromodomain inhibitors (BRD9i) or a degrader (dBRD9) led to reduction in a number of interferon-stimulated genes (ISGs) following stimulation with endotoxin lipid A. BRD9-dependent genes overlapped highly with a subset of genes differentially regulated by BET protein inhibition with JQ1 following endotoxin stimulation. We find that the BET protein BRD4 is cobound with BRD9 in unstimulated macrophages and corecruited upon stimulation to ISG promoters along with STAT1, STAT2, and IRF9, components of the ISGF3 complex activated downstream of IFN-alpha receptor stimulation. In the presence of BRD9i or dBRD9, STAT1-, STAT2-, and IRF9-binding is reduced, in some cases with reduced binding of BRD4. These results demonstrate a specific role for BRD9 and the ncBAF complex in ISG activation and identify an activity for BRD9 inhibitors and degraders in dampening endotoxin- and IFN-dependent gene expression.

Inflammation in the Pathogenesis of Arrhythmogenic Cardiomyopathy: Secondary Event or Active Driver?

Arrhythmogenic cardiomyopathy (ACM) is a rare inherited cardiac disease characterized by arrhythmia and progressive fibro-fatty replacement of the myocardium, which leads to heart failure and sudden cardiac death. Inflammation contributes to disease progression, and it is characterized by inflammatory cell infiltrates in the damaged myocardium and inflammatory mediators in the blood of ACM patients. However, the molecular basis of inflammatory process in ACM remains under investigated and it is unclear whether inflammation is a primary event leading to arrhythmia and myocardial damage or it is a secondary response triggered by cardiomyocyte death. Here, we provide an overview of the proposed players and triggers involved in inflammation in ACM, focusing on those studied using in vivo and in vitro models. Deepening current knowledge of inflammation-related mechanisms in ACM could help identifying novel therapeutic perspectives, such as anti-inflammatory therapy.

Modelling novelty detection in the thalamocortical loop

In complex natural environments, sensory systems are constantly exposed to a large stream of inputs. Novel or rare stimuli, which are often associated with behaviorally important events, are typically processed differently than the steady sensory background, which has less relevance. Neural signatures of such differential processing, commonly referred to as novelty detection, have been identified on the level of EEG recordings as mismatch negativity and the level of single neurons as stimulus-specific adaptation. Here, we propose a multi-scale recurrent network with synaptic depression to explain how novelty detection can arise in the whisker-related part of the somatosensory thalamocortical loop. The architecture and dynamics of the model presume that neurons in cortical layer 6 adapt, via synaptic depression, specifically to a frequently presented stimulus, resulting in reduced population activity in the corresponding cortical column when compared with the population activity evoked by a rare stimulus. This difference in population activity is then projected from the cortex to the thalamus and amplified through the interaction between neurons of the primary and reticular nuclei of the thalamus, resulting in spindle-like, rhythmic oscillations. These differentially activated thalamic oscillations are forwarded to cortical layer 4 as a late secondary response that is specific to rare stimuli that violate a particular stimulus pattern. Model results show a strong analogy between this late single neuron activity and EEG-based mismatch negativity in terms of their common sensitivity to presentation context and timescales of response latency, as observed experimentally. Our results indicate that adaptation in L6 can establish the thalamocortical dynamics that produce signatures of SSA and MMN and suggest a mechanistic model of novelty detection that could generalize to other sensory modalities.

Attenuation of the Airway and Cardiovascular Responses to Extubation in Chronic Smokers by Prior Treatment with Dexmedetomidine, Fentanyl, and their Combination

BACKGROUND: Respiratory complications and hemodynamic changes during and after extubation are more common than during tracheal intubation and induction of anesthesia. AIM: The objective of this study was to compare the efficacy of prior treatment with dexmedetomidine, fentanyl, and their combination on the attenuation of the airway and cardiovascular responses to extubation. METHODS: The subjects were adult chronic male smokers, representing the population in which secondary response to extubation is most common. A randomized double-blinded comparative trial was conducted on 66 patients who were 20–60 years of age, chronic male smokers, scheduled for elective surgeries, and divided into three equal groups according to given drug 20 min before the end of surgery. Group A (n = 22) received 1 ug. kg−1 dexmedetomidine, Group B (n = 22) received fentanyl 1 ug. kg−1, and Group C (n = 22) received a mixture of the previously used drugs in the same doses. Time to and quality of extubation, airway and hemodynamic responses, and post-operative agitation and sedation were recorded. RESULTS: Hemodynamic responses and quality of extubation were better in both Groups A and C than patients in Group B at the expense of increasing time to extubation, post-extubation sedation, and delayed recovery in Group C. CONCLUSION: Single-dose dexmedetomidine 1 ug. kg-1 given 15 min before extubation in chronic cigarette smokers provided better attenuation of the airway and cardiovascular responses to extubation and suctioning with better recovery profile when compared to fentanyl 1 ug. kg-1 and dexmedetomidine mixed with fentanyl in the same previous doses.

Novel chemotherapeutic agent FX-9 activates NF-κB signaling and induces G1 phase arrest by activating CDKN1A in a human prostate cancer cell line

Background The aminoisoquinoline FX-9 shows pro-apoptotic and antimitotic effects against lymphoblastic leukemia cells and prostate adenocarcinoma cells. In contrast, decreased cytotoxic effects against non-neoplastic blood cells, chondrocytes, and fibroblasts were observed. However, the actual FX-9 molecular mode of action is currently not fully understood. Methods In this study, microarray gene expression analysis comparing FX-9 exposed and unexposed prostate cancer cells (PC-3 representing castration-resistant prostate cancer), followed by pathway analysis and gene annotation to functional processes were performed. Immunocytochemistry staining was performed with selected targets. Results Expression analysis revealed 0.83% of 21,448 differential expressed genes (DEGs) after 6-h exposure of FX-9 and 0.68% DEGs after 12-h exposure thereof. Functional annotation showed that FX-9 primarily caused an activation of inflammatory response by non-canonical nuclear factor-kappa B (NF-κB) signaling. The 6-h samples showed activation of the cell cycle inhibitor CDKN1A which might be involved in the secondary response in 12-h samples. This secondary response predominantly consisted of cell cycle-related changes, with further activation of CDKN1A and inhibition of the transcription factor E2F1, including downstream target genes, resulting in G1-phase arrest. Matching our previous observations on cellular level senescence signaling pathways were also found enriched. To verify these results immunocytochemical staining of p21 Waf1/Cip1 (CDKN1A), E2F1 (E2F1), PAI-1 (SERPNE1), and NFkB2/NFkB p 100 (NFKB2) was performed. Increased expression of p21 Waf1/Cip1 and NFkB2/NFkB p 100 after 24-h exposure to FX-9 was shown. E2F1 and PAI-1 showed no increased expression. Conclusions FX-9 induced G1-phase arrest of PC-3 cells through activation of the cell cycle inhibitor CDKN1A, which was initiated by an inflammatory response of noncanonical NF-κB signaling.

Activated PI3Kδ signals compromise plasma cell survival via limiting autophagy and increasing ER stress

While phosphatidylinositide 3-kinase delta (PI3Kδ) plays a critical role in humoral immunity, the requirement for PI3Kδ signaling in plasma cells remains poorly understood. Here, we used a conditional mouse model of activated PI3Kδ syndrome (APDS), to interrogate the function of PI3Kδ in plasma cell biology. Mice expressing a PIK3CD gain-of-function mutation (aPIK3CD) in B cells generated increased numbers of memory B cells and mounted an enhanced secondary response but exhibited a rapid decay of antibody levels over time. Consistent with these findings, aPIK3CD expression markedly impaired plasma cell generation, and expression of aPIK3CD intrinsically in plasma cells was sufficient to diminish humoral responses. Mechanistically, aPIK3CD disrupted ER proteostasis and autophagy, which led to increased plasma cell death. Notably, this defect was driven primarily by elevated mTORC1 signaling and modulated by treatment with PI3Kδ-specific inhibitors. Our findings establish an essential role for PI3Kδ in plasma cell homeostasis and suggest that modulating PI3Kδ activity may be useful for promoting and/or thwarting specific immune responses.

The Response and Feedback of Ocean Mesoscale Eddies to Four Sequential Typhoons in 2016 Based on Multiple Satellite Observations and Argo Floats

Four sequential tropical cyclones generated and developed in the Northwest Pacific Ocean (NWP) in 2014, which had significant impacts on the oceanic environment and coastal regions. Based on a substantial dataset of multiple-satellite observations, Argo profiles, and reanalysis data, we comprehensively investigated the interactions between the oceanic environment and sequential tropical cyclones. Super typhoon Neoguri (2014) was the first typhoon-passing studied area, with the maximum sustained wind speed of 140 kts, causing strong cold wake along the track. The location of the strongest cold wake was consistent with the pre-existing cyclonic eddy (CE), in which the average sea surface temperature (SST) cooling exceeded −5 °C. Subsequently, three tropical cyclones passed the ocean environment left by Neoguri, namely, the category 2 typhoon Matmo (2014), the tropical cyclone Nakri (2014) and the category 5 typhoon Halong (2014), which caused completely different subsequent responses. In the CE, due to the fact that the ocean stratification was strongly destroyed by Neoguri and difficult to recover, even the weak Nakri could cause a secondary response, but the secondary SST cooling would be overridden by the first response and thus could cause no more serious ocean disasters. If the subsequent typhoon was super typhoon Halong, it could cause an extreme secondary SST cooling, exceeding −8 °C, due to the deep upwelling, exceeding 700 m, surpassing the record of the maximum cooling caused by the first typhoon. In the anti-cyclonic eddy (AE), since the first typhoon Neoguri caused strong seawater mixing, it was difficult for the subsequent weak typhoons to mix the deeper, colder and saltier water into the surface, thus inhibiting secondary SST cooling, and even the super typhoon Halong would only cause as much SST cooling as the first typhoon. Therefore, the ocean responses to sequential typhoons depended on not only TCs intensity, but also TCs track order and ocean mesoscale eddies. In turn, the cold wake caused by the first typhoon, Neoguri, induced different feedback effects on different subsequent typhoons.

Personalised Profiling of Innate Immune Memory Induced by Nano-Imaging Particles in Human Monocytes

Engineered nanoparticles used for medical purposes must meet stringent safety criteria, which include immunosafety, i.e., the inability to activate possibly detrimental immune/inflammatory effects. Even medical nanomaterials devoid of direct immunotoxic or inflammatory effects may have an impact on human health if able to modify innate memory, which is the ability to “prime” future immune responses towards a different, possibly more detrimental reactivity. Although innate memory is usually protective, anomalous innate memory responses may be at the basis of immune pathologies. In this study, we have examined the ability of two nanomaterials commonly used for diagnostic imaging purposes, gold and iron oxide nanoparticles, to induce or modulate innate memory, using an in vitro model based on human primary monocytes. Monocytes were exposed in culture to nanoparticles alone or together with the bacterial agent LPS (priming phase/primary response), then rested for six days (extinction phase), and eventually challenged with LPS (memory/secondary response). The memory response to the LPS challenge was measured as changes in the production of inflammatory (TNFα, IL-6) and anti-inflammatory cytokines (IL-10, IL-1Ra), as compared to unprimed monocytes. The results show that both types of nanoparticles can have an effect in the induction of memory, with changes observed in the cytokine production. By comparing nanomaterials of different shapes (spherical vs. rod-shaped gold particles) and different size (17 vs. 22 nm diameter spherical iron oxide particles), it was evident that innate memory could be differentially induced and modulated depending on size, shape and chemical composition. However, the main finding was that the innate memory effect of the particles was strongly donor-dependent, with monocytes from each donor showing a distinct memory profile upon priming with the same particles, thereby making impossible to draw general conclusions on the particle effects. Thus, in order to predict the effect of imaging nanoparticles on the innate memory of patients, a personalised profiling would be required, able to take in consideration the peculiarities of the individual innate immune reactivity.

Clavibacter michiganensis Downregulates Photosynthesis and Modifies Monolignols Metabolism Revealing a Crosstalk with Tomato Immune Responses

The gram-positive pathogenic bacterium Clavibacter michiganensis subsp. michiganensis (Cmm) causes bacterial canker disease in tomato, affecting crop yield and fruit quality. To understand how tomato plants respond, the dynamic expression profile of host genes was analyzed upon Cmm infection. Symptoms of bacterial canker became evident from the third day. As the disease progressed, the bacterial population increased in planta, reaching the highest level at six days and remained constant till the twelfth day post inoculation. These two time points were selected for transcriptomics. A progressive down-regulation of key genes encoding for components of the photosynthetic apparatus was observed. Two temporally separated defense responses were observed, which were to an extent interdependent. During the primary response, genes of the phenylpropanoid pathway were diverted towards the synthesis of monolignols away from S-lignin. In dicots, lignin polymers mainly consist of G- and S-units, playing an important role in defense. The twist towards G-lignin enrichment is consistent with previous findings, highlighting a response to generate an early protective barrier and to achieve a tight interplay between lignin recomposition and the primary defense response mechanism. Upon progression of Cmm infection, the temporal deactivation of phenylpropanoids coincided with the upregulation of genes that belong in a secondary response mechanism, supporting an elegant reprogramming of the host transcriptome to establish a robust defense apparatus and suppress pathogen invasion. This high-throughput analysis reveals a dynamic reorganization of plant defense mechanisms upon bacterial infection to implement an array of barriers preventing pathogen invasion and spread.

Influence of portal hypertension on the state of the hemomicrocirculatory level of rat skin: morphological study

The effect of portal hypertension on the condition of the hemomicrocirculatory tract of the skin male white nonlinear rats was studied. Hypertension was simulated by applying a ligature to the portal vein according to the conventional method. After 30, 60 and 90 days, skin biopsies were taken from the anterior abdominal wall for optical microscopy examination, which was subjected to routine histological processing. In animals of the control group, which underwent sham surgery, the skin remained unchanged. In rats of the experimental group after 30 days in the basal layer of the epidermis there were cells with signs of degenerative changes. The basement membrane of the epidermis had small areas with thinning and thickening, which in both cases were accompanied by the formation of small and massive leukocyte infiltrates. Its edema was noted, which extended to the papillary and reticular layer of the dermis. Elastic fibers were unchanged. Only 50% of the microvessels looked intact, 20% microvessels were narrowed, and 30% had signs of stagnant hyperemia due to dilatation. After 60 days the epidermis was thinned, in the basal layer the cells with picnotized and displaced to the periphery nucleus were detected. In all studied layers of epidermis, besides reticular layer, expansion of arterioles, venules and capillary loops was observed. After 90 days the epidermis was represented by 3-6 layers of cells, its general refinement was observed due to the reduction of the thickness of the spinous and granular layers with simultaneous thickening of the horny layer. In the papillary layer of the dermis, the number of thin-walled blood vessels was increased. The average diameter of the arterioles of the hypodermis decreased relative to the values on the 60th day. The mean diameter of the arterioles of the subdermal network on the border of the papillary and reticular layers and capillary loops under the basal membrane was reduced what can be a sign of sclerotic changes in the microvessels. The venules diameter remained increased. In arterioles and venules, stasis of the microvessels, edema of their walls, microaneurysm, and venous sacculation were detected. Thus, we believe that the changes in the microvessels of the skin, which are observed in portal hypertension, are the result of hemodynamic disturbances in the tissue, which is a complex chain of hemomicrocirculatory flow to the direct action of hypertension in the portal vein and is a secondary response to systemic vascular damage.