Alpha Activity During Lucid Dreaming

We have been interested in the electrophysiological correlates of lucid dreaming (LD) since early work in this laboratory suggested a relationship between lucidity and alpha activity (Ogilvie, Hunt, Sawicki &McGowan, 1978; Ogilvie, Hunt, Tyson, Lucescu & Jeakins, 1982; Tyson, Ogilvie, & Hunt, 1984). Until now, this alpha-lucidity hypothesis had not been tested in our lab on high frequency lucid dreamers who signal while in REM sleep, and LaBerge (1980; 1981) has not observed any changes in alpha in signalled episodes of lucidity. The present report describes computer analyses of EEG activity obtained during eight laboratory nights from a frequent lucid dreamer. In addition to standard polysomnographic measures, CCTV monitoring with two cameras and a screen splitter permitted simultaneous display and videotaping of the subject's (RJS's) face and concurrent polygraphic activity. This was particularly useful during REM, since RJS had trained himself to sleep on his back and to try to signal from that position. In the month prior to the lab nights, RJS spend over an hour per day meditating. He also used LaBerge's lucidity induction or cognitive self-instruction techniques and reported LD rates in excess of one per night. He practiced signalling (using three blinks/ rapid vertical eye movements) and thought he had successfully signalled several LDs while at home

Blood pressure–associated polymorphism controls ARHGAP42 expression via serum response factor DNA binding

We recently demonstrated that selective expression of the Rho GTPase-activating protein ARHGAP42 in smooth muscle cells (SMCs) controls blood pressure by inhibiting RhoA-dependent contractility, providing a mechanism for the blood pressure–associated locus within the ARHGAP42 gene. The goals of the current study were to identify polymorphisms that affect ARHGAP42 expression and to better assess ARHGAP42’s role in the development of hypertension. Using DNase I hypersensitivity methods and ENCODE data, we have identified a regulatory element encompassing the ARHGAP42 SNP rs604723 that exhibits strong SMC-selective, allele-specific activity. Importantly, CRISPR/Cas9–mediated deletion of this element in cultured human SMCs markedly reduced endogenous ARHGAP42 expression. DNA binding and transcription assays demonstrated that the minor T allele variation at rs604723 increased the activity of this fragment by promoting serum response transcription factor binding to a cryptic cis-element. ARHGAP42 expression was increased by cell stretch and sphingosine 1-phosphate in a RhoA-dependent manner, and deletion of ARHGAP42 enhanced the progression of hypertension in mice treated with DOCA-salt. Our analysis of a well-characterized cohort of untreated borderline hypertensive patients suggested that ARHGAP42 genotype has important implications in regard to hypertension risk. Taken together, our data add insight into the genetic mechanisms that control blood pressure and provide a potential target for individualized antihypertensive therapies

Blood pressure–associated polymorphism controls ARHGAP42 expression via serum response factor DNA binding

We recently demonstrated that selective expression of the Rho GTPase-activating protein ARHGAP42 in smooth muscle cells (SMCs) controls blood pressure by inhibiting RhoA-dependent contractility, providing a mechanism for the blood pressure-associated locus within the ARHGAP42 gene. The goals of the current study were to identify polymorphisms that affect ARHGAP42 expression and to better assess ARHGAP42's role in the development of hypertension. Using DNase I hypersensitivity methods and ENCODE data, we have identified a regulatory element encompassing the ARHGAP42 SNP rs604723 that exhibits strong SMC-selective, allele-specific activity. Importantly, CRISPR/Cas9-mediated deletion of this element in cultured human SMCs markedly reduced endogenous ARHGAP42 expression. DNA binding and transcription assays demonstrated that the minor T allele variation at rs604723 increased the activity of this fragment by promoting serum response transcription factor binding to a cryptic cis-element. ARHGAP42 expression was increased by cell stretch and sphingosine 1-phosphate in a RhoA-dependent manner, and deletion of ARHGAP42 enhanced the progression of hypertension in mice treated with DOCA-salt. Our analysis of a well-characterized cohort of untreated borderline hypertensive patients suggested that ARHGAP42 genotype has important implications in regard to hypertension risk. Taken together, our data add insight into the genetic mechanisms that control blood pressure and provide a potential target for individualized antihypertensive therapies

Blood pressure–associated polymorphism controls ARHGAP42 expression via serum response factor DNA binding

We recently demonstrated that selective expression of the Rho GTPase-activating protein ARHGAP42 in smooth muscle cells (SMCs) controls blood pressure by inhibiting RhoA-dependent contractility, providing a mechanism for the blood pressure–associated locus within the ARHGAP42 gene. The goals of the current study were to identify polymorphisms that affect ARHGAP42 expression and to better assess ARHGAP42’s role in the development of hypertension. Using DNase I hypersensitivity methods and ENCODE data, we have identified a regulatory element encompassing the ARHGAP42 SNP rs604723 that exhibits strong SMC-selective, allele-specific activity. Importantly, CRISPR/Cas9–mediated deletion of this element in cultured human SMCs markedly reduced endogenous ARHGAP42 expression. DNA binding and transcription assays demonstrated that the minor T allele variation at rs604723 increased the activity of this fragment by promoting serum response transcription factor binding to a cryptic cis-element. ARHGAP42 expression was increased by cell stretch and sphingosine 1-phosphate in a RhoA-dependent manner, and deletion of ARHGAP42 enhanced the progression of hypertension in mice treated with DOCA-salt. Our analysis of a well-characterized cohort of untreated borderline hypertensive patients suggested that ARHGAP42 genotype has important implications in regard to hypertension risk. Taken together, our data add insight into the genetic mechanisms that control blood pressure and provide a potential target for individualized antihypertensive therapies

MALT1 Protease Activity Is Required for Innate and Adaptive Immune Responses

<div><p>CARMA-BCL10-MALT1 signalosomes play important roles in antigen receptor signaling and other pathways. Previous studies have suggested that as part of this complex, MALT1 functions as both a scaffolding protein to activate NF-κB through recruitment of ubiquitin ligases, and as a protease to cleave and inactivate downstream inhibitory signaling proteins. However, our understanding of the relative importance of these two distinct MALT1 activities has been hampered by a lack of selective MALT1 protease inhibitors with suitable pharmacologic properties. To fully investigate the role of MALT1 protease activity, we generated mice homozygous for a protease-dead mutation in MALT1. We found that some, but not all, MALT1 functions in immune cells were dependent upon its protease activity. Protease-dead mice had defects in the generation of splenic marginal zone and peritoneal B1 B cells. CD4⁺ and CD8⁺ T cells displayed decreased T cell receptor-stimulated proliferation and IL-2 production while B cell receptor-stimulated proliferation was partially dependent on protease activity. In dendritic cells, stimulation of cytokine production through the Dectin-1, Dectin-2, and Mincle C-type lectin receptors was also found to be partially dependent upon protease activity. <i>In vivo</i>, protease-dead mice had reduced basal immunoglobulin levels, and showed defective responses to immunization with T-dependent and T-independent antigens. Surprisingly, despite these decreased responses, MALT1 protease-dead mice, but not MALT1 null mice, developed mixed inflammatory cell infiltrates in multiple organs, suggesting MALT1 protease activity plays a role in immune homeostasis. These findings highlight the importance of MALT1 protease activity in multiple immune cell types, and in integrating immune responses <i>in vivo</i>.</p></div

MALT1 protease-dead mice develop inflammatory cell infiltrates in multiple organs.

<p>H&E-stained histological sections of tissues from MALT1 <i>Wt</i> (left panels A, C, E, G; n = 3) and <i>Malt1</i>^PD/PD (right panels B, D, F H; n = 6) mice. Results from <i>Malt1</i>^-/- (n = 3) and <i>Malt1</i>^+/PD (n = 2) mice were identical to <i>Wt</i> mice. (A, B) Lung at 100X magnification. Airways of <i>Wt</i> and <i>Malt1</i>^PD/PD mice, including the B, bronchiolar (B) and terminal airways (TA) were relatively clear of inflammatory infiltrates. Distinctive findings were present in the lungs of <i>Malt1</i>^PD/PD mice, which featured pulmonary veins (V) surrounded by a prominent mixed mononuclear inflammatory cell infiltrate predominated by lymphocytes and histiocytic cells (inset, 400X magnification). (C, D) Limbic region of eye (150X magnification). Cornea from <i>Wt</i> eye demonstrates typical anatomical features of a thin corneal epithelium (CE) overlying the substantia propria (SP). Occasional linear nuclei in the SP are part of the normal fibroblast populations. Sclera (S), pigmented iris (I) and lens (L) are normal in <i>Malt1</i>^PD/PD mice. Cornea from <i>Malt1</i>^PD/PD mice consistently had neutrophilic inflammatory cell infiltrates (arrows) of varying severity in the substantia propria of the cornea. This particular <i>Malt1</i>^PD/PD mouse also had a florid neutrophilic infiltrate towards the central portion of the cornea (inset, 150X magnification). (E, F) Non-glandular stomach (50X magnification). Normal non-glandular stomach was comprised of stratified squamous epithelium (Ep) with a small amount of keratinized stratum corneum (Sc) and only occasional inflammatory cells evident in the submucosa (Sub). Non-glandular stomach from <i>Malt1</i>^PD/PD mice demonstrated marked hyperkeratosis of the stratum corneum and prominent hyperplasia of the epithelium. A mixed inflammatory infiltrate was commonly present in the mural wall, with this particular mouse having a moderate neutrophilic infiltrate in the submucosa (arrow). (G, H) Glandular stomach (50X magnification). The fundic mucosa (Muc) of the <i>Wt</i> mouse is regular, even, and is formed by undulating gastric pits. The relatively acellular tunica adventitia (Ta) is the thin tissue layer (arrow) sandwiched between the thicker tunica muscularis (Tm) and the adipose tissue (Ad) / pancreas (Pan). Reactive hyperplasia of the glandular mucosa was present in <i>Malt1</i>^PD/PD mice, and characterized by prominent thickening of the mucosa and loss of regular and even mucosal architecture. Mixed inflammatory cell infiltrates were increased in the mural wall of stomachs from <i>Malt1</i>^PD/PD mice, with this particular mouse demonstrating an intense inflammatory infiltrate of the tunica adventicia that is predominated by populations of lymphocytes and histiocytic cells.</p

Mixed inflammatory infiltrates of T cells, B cells, and macrophages in MALT1 protease-dead mice.

<p>IHC photomicrographs (80X magnification, insets 400X magnification) of inflammatory cell infiltrates surrounding vessels (V) of the lung and tunica adventicia of glandular stomach in <i>Malt1</i>^PD/PD mice, using antibodies against: (A, D) CD3 to detect T cells; (B, E) CD45 to detect B-cell; and (C, F) F4/80 to detect macrophage. Non-selective background staining was higher with the F4/80 antibody, as evident with the pale staining of erythrocytes within the vessel lumen (V, Fig 8C). Therefore, macrophage identification was made by considering the higher staining intensity and morphology of cells stained (inset, Fig 8C). IHC photomicrographs of inflammatory cell infiltrates in lung of <i>Malt1</i>^PD/PD mice (8A, C) demonstrates a mixed population of T-cells, B-cells and macrophages comprising the perivascular cuffs of pulmonary vessels. Similar to the lung infiltrates of <i>Malt1</i>^PD/PD, inflammatory infiltrates in deeper glandular portions of stomach had mixed population of T cells, B cells and macrophages in the tunica adventicia (Fig 8D, F).</p