Asthma is a complex disease that is promoted by dysregulated immunity and the presence of many cytokine and lipid mediators. Despite this, there is a paucity of data demonstrating the combined ...effects of multiple mediators in asthma pathogenesis. Group 2 innate lymphoid cells (ILC2s) have recently been shown to play important roles in the initiation of allergic inflammation; however, it is unclear whether lipid mediators, such as cysteinyl leukotrienes (CysLTs), which are present in asthma, could further amplify the effects of IL-33 on ILC2 activation and lung inflammation. In this article, we show that airway challenges with the parent CysLT, leukotriene C4 (LTC4), given in combination with low-dose IL-33 to naive wild-type mice, led to synergistic increases in airway Th2 cytokines, eosinophilia, and peribronchial inflammation compared with IL-33 alone. Further, the numbers of proliferating and cytokine-producing lung ILC2s were increased after challenge with both LTC4 and IL-33. Levels of CysLT1R, CysLT2R, and candidate leukotriene E4 receptor P2Y12 mRNAs were increased in ILC2s. The synergistic effect of LTC4 with IL-33 was completely dependent upon CysLT1R, because CysLT1R
mice, but not CysLT2R
mice, had abrogated responses. Further, CysLTs directly potentiated IL-5 and IL-13 production from purified ILC2s stimulated with IL-33 and resulted in NFAT1 nuclear translocation. Finally, CysLT1R
mice had reduced lung eosinophils and ILC2 responses after exposure to the fungal allergen
Thus, CysLT1R promotes LTC4- and
-induced ILC2 activation and lung inflammation. These findings suggest that multiple pathways likely exist in asthma to activate ILC2s and propagate inflammatory responses.
ILC2 numbers positively correlated with mean eosinophils per hpf (r = 0.7; P < .0001; CI, 0.45-0.84; Pearson r correlation) enumerated by independent pathologic analysis of hematoxylin and eosin ...sections (Fig 2, B). ...esophageal ILC2s correlated strongly with eosinophilia and were increased in samples from patients with active EoE compared with patients with inactive EoE, PPI-REE, and control non-EoE tissue.
Because PGD2 is released by several cell types important to allergic inflammation including mast cells, macrophages, and eosinophils, activation of these cell types at mucosal sites in the upper or ...lower airway could promote PGD2-mediated chemotaxis in tissues.
We developed an index to measure progressive realization for the human right to water and sanitation. While in this study we demonstrate its application to the non-discrimination and equality ...component for water, the conceptual approach of the index can be used for all the different components of the human right. The index was composed of one structural, one process, and two outcome indicators and is bound between −1 and 1, where negative values indicate regression and positive values indicate progressive realization. For individual structural and process indicators, only discrete values such as −1, −0.5, 0, 0.5, and 1 were allowed. For the outcome indicators, any value between −1 and 1 was possible, and a State's progress was evaluated using rates of change. To create an index that would allow for fair comparisons between States and across time, these rates of change were compared to benchmarked rates, which reflect the maximum rates a State can achieve. Using this approach, we calculated the index score for 56 States in 2010 for which adequate data were available and demonstrated that these index scores were not dependent on factors such as achieved level of coverage or gross national income. The proposed index differs from existing measures of inequality as it measures rate of change and not level of achievement, and thus addresses the principle of progressive realization that is fundamental to human rights.
Connexin-43 (Cx43) gap junctions provide intercellular coupling, which ensures rapid action potential propagation and synchronized heart contraction. Alterations in Cx43 localization and reductions ...in gap junction coupling occur in failing hearts, contributing to ventricular arrhythmias and sudden cardiac death. Recent reports have found that an internally translated Cx43 isoform, GJA1-20k, is an auxiliary subunit for the trafficking of Cx43 in heterologous expression systems. Here, we have created a mouse model by using CRISPR technology to mutate a single internal translation initiation site in Cx43 (M213L mutation), which generates full-length Cx43, but not GJA1-20k. We found that GJA1M213L/M213L mice had severely abnormal electrocardiograms despite preserved contractile function, reduced total Cx43, and reduced gap junctions, and they died suddenly at 2 to 4 weeks of age. Heterozygous GJA1M213L/WT mice survived to adulthood with increased ventricular ectopy. Biochemical experiments indicated that cytoplasmic Cx43 had a half-life that was 50% shorter than membrane-associated Cx43. Without GJA1-20k, poorly trafficked Cx43 was degraded. The data support that GJA1-20k, an endogenous entity translated independently of Cx43, is critical for Cx43 gap junction trafficking, maintenance of Cx43 protein, and normal electrical function of the mammalian heart.
Abstract Group 2 innate lymphoid cells (ILC2s) have recently been identified in human nasal polyps, but whether numbers of ILC2s differ by polyp endotype or are influenced by corticosteroid use is ...unknown. Here, we show that eosinophilic nasal polyps contained double the number of ILC2s vs. non-eosinophilic polyps. Polyp ILC2s were also reduced by 50% in patients treated with systemic corticosteroids. Further, using a fungal allergen challenge mouse model, we detected greatly reduced Th2 cytokine-producing and Ki-67 + proliferating lung ILC2s in mice receiving dexamethasone. Finally, ILC2 Annexin V staining revealed extensive apoptosis after corticosteroid treatment in vivo and in vitro . Thus, ILC2s are elevated in the eosinophilic nasal polyp endotype and systemic corticosteroid treatment correlated with reduced polyp ILC2s. Finally, allergen-challenged mice showed reduced ILC2s and increased ILC2 apoptosis after corticosteroid treatment suggesting that ILC2 may be responsive to corticosteroids in eosinophilic respiratory disease.
The objective of this study was to use a supervised machine learning (ML) platform and a national dataset to identify factors important in classifying common types of dizziness.
Using established ...clinical criteria and responses to the balance and dizziness supplement from the 2016 National health Interview Survey (n = 33,028), case definitions for vestibular migraine (VM), benign paroxysmal positional vertigo (BPPV) Ménière's disease (MD), persistent postural-perceptual dizziness (PPPD), superior canal dehiscence (SCD), and bilateral vestibular hypofunction (BVH) were generated. One hundred thirty-six variables consisting of sociodemographic characteristics and medical comorbidities were used to develop decision tree models to predict these common types of dizziness.
The one-year prevalence of dizziness in the U.S. was 16.8% (5562 respondents). VM was highly prevalent, representing 4.0% of the overall respondents (n = 1327). ML decision tree models were able to correctly classify all 6 dizziness subtypes with high accuracy (sensitivity range, 70–92%; specificity range, 89–99%) using responses to questions about functional limitations due to dizziness, such as falls due to dizziness and modification of social activities due to dizziness.
In a large population-based dataset, supervised ML models accurately predicted dizziness subtypes according to responses to questions that do not pertain to dizziness symptoms alone.
Innate lymphoid cells (ILC) promote lung inflammation in asthma through cytokine production. RNA-binding proteins (RBPs) are critical post-transcriptional regulators, although less is known about ...RBPs in ILC biology. Here, we demonstrate that RNA-binding motif 3 (RBM3) is highly expressed in lung ILCs and is further induced by alarmins TSLP and IL-33. Rbm3
and Rbm3
Rag2
mice exposed to asthma-associated Alternaria allergen develop enhanced eosinophilic lung inflammation and ILC activation. IL-33 stimulation studies in vivo and in vitro show that RBM3 suppressed lung ILC responses. Further, Rbm3
ILCs from bone marrow chimeric mice display increased ILC cytokine production suggesting an ILC-intrinsic suppressive function of RBM3. RNA-sequencing of Rbm3
lung ILCs demonstrates increased expression of type 2/17 cytokines and cysteinyl leukotriene 1 receptor (CysLT1R). Finally, Rbm3
Cyslt1r
mice show dependence on CysLT1R for accumulation of ST2
IL-17
ILCs. Thus, RBM3 intrinsically regulates lung ILCs during allergen-induced type 2 inflammation that is partially dependent on CysLT1R.
CRISPR/Cas9-mediated transcriptional interference (CRISPRi) enables programmable gene knock-down, yielding loss-of-function phenotypes for nearly any gene. Effective, inducible CRISPRi has been ...demonstrated in budding yeast, and genome-scale guide libraries enable systematic, genome-wide genetic analysis.
We present a comprehensive yeast CRISPRi library, based on empirical design rules, containing 10 distinct guides for most genes. Competitive growth after pooled transformation revealed strong fitness defects for most essential genes, verifying that the library provides comprehensive genome coverage. We used the relative growth defects caused by different guides targeting essential genes to further refine yeast CRISPRi design rules. In order to obtain more accurate and robust guide abundance measurements in pooled screens, we link guides with random nucleotide barcodes and carry out linear amplification by in vitro transcription.
Taken together, we demonstrate a broadly useful platform for comprehensive, high-precision CRISPRi screening in yeast.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
To the Editor: Type 2 innate lymphoid cells (ILC2s) have recently been discovered and include nuocytes, natural helper cells, and innate type 2 helper cells.1,2 ILC2s do not express known surface ...markers for T, B, natural killer, or natural killer T cells (lineage-negative) and produce large amounts of TH2 cytokines in response to cytokines IL-33, IL-25, thymic stromal lymphopoietin, and leukotriene D4.2-4 Studies in animal models have shown that ILC2s contribute to allergen-induced airway hyperresponsiveness and type 2 lung inflammatory responses, suggesting that ILC2s may have a role in asthma and allergic disease.2 Human ILC2s have been detected in peripheral blood, gastrointestinal tract, lung, bronchoalveolar lavage, and nasal polyps and are defined as lineage-negative lymphocytes that express the chemoattractant receptor homologous molecule expressed on TH2 lymphocytes (CRTH2).5 We have previously reported that peripheral blood ILC2s highly express the master TH2 cytokine transcription factor GATA-binding protein 3, supporting a role for rapid ILC2 TH2 cytokine production.6 Despite the potential for ILC2s to contribute to human allergic disease, no studies have yet reported whether allergen exposure in sensitized allergic rhinitis subjects has any effect on peripheral blood ILC2 levels. ...we hypothesized that CRTH2+ ILC2s may be increased in the peripheral blood of cat-allergic individuals after nasal cat allergen challenge. Human ILC2s are a recently discovered population of cells that are known to express CRTH2 and CD161,5 but reported expression of other markers is very limited and we thus sought to identify expression of novel human ILC2 surface molecules. Because mouse ILC2s express molecules involved in T-cell-B-cell interactions including Inducible T-cell Costimulator,2 we assessed ILC2s for levels of CD84, a CD2 signaling lymphocyte activation molecule family member with a role in T-cell-B-cell contact.7 Interestingly, we detected high levels of CD84 on peripheral blood ILC2s from cat-allergic individuals at baseline (Fig 1, B).
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