Sufang Yang, Alexander Dizhoor, David J. Wilson, Grazyna Adamus; GCAP1, Rab6, and HSP27: Novel Autoantibody Targets in Cancer-Associated Retinopathy and Autoimmune Retinopathy. Trans. Vis. Sci. Tech. 2016;5(3):1. doi: 10.1167/tvst.5.3.1.
Purpose: Autoantibodies (AAbs) with different retinal specificities were reported in cancer-associated retinopathy (CAR) and autoimmune retinopathy (AR). The goal was to identify the small retinal proteins of apparent molecular mass of 23-kDa often recognized by patients' AAbs.
Methods: Sera specific for a 23-kDa retinal protein of 173 patients were investigated retrospectively by Western blotting and double immunofluorescence confocal microscopy. A proteomic analysis revealed new 23-kDa protein candidates, including guanylyl cyclase-activating proteins (GCAPs), heat shock protein 27 (HSP27), and Rab6A GTPase (Rab6A).
Results: Among the cohort of 173 patients, only 68 had anti-recoverin AAbs and the remaining 105 reacted with 4 unique proteins, which were identified as a Rab6A, HSP27, GCAP1, and GCAP2. Confocal images from a double labeling study confirmed the reactivity of AAbs with different types of cells in human retina, consistent with the target protein's respective cellular functions. Patients (62/173) had been diagnosed with various kinds of cancer, including 20% of patients who had anti-recoverin, 11% anti-Rab6A, and 5% anti-HSP27 AAbs. Only 50% of recoverin-seropositive patients had cancer and the individuals with anti-recoverin AAbs had a significantly higher likelihood to be diagnosed with cancer than patients with other anti-23-kDa AAbs.
Conclusions: The newly discovered retinal autoantigens may be involved in pathogenicity of CAR and AR. The recognition of AAbs against various retinal proteins associated with autoimmune retinal degeneration broadens the group of proteins related with these entities.
Translational Relevance: Patients with anti-recoverin, anti-GCAP1, anti-Rab6A, and anti-HSP27 AAbs represented diverse clinical phenotypes, so the presence of disease-associated AAbs provides important information for molecular diagnosis.
Cancer-associated retinopathy (CAR) and autoimmune retinopathy (AR) are rare conditions that are immunologically heterogeneous, produce a number of different symptoms, and are associated with a diverse anti-retinal autoantibody repertoire.1–6 Autoimmune retinopathy resembles CAR but patients with AR do not have diagnosed cancer at the initial ocular exam, or the cancer may be in too early a stage of development to be detected by conventional methods.7 However, tumors may be detected in subsequent years with the retinopathy and autoantibodies (AAbs) preceding diagnosis of cancer by years.8 The specificities of antiretinal AAbs range from photoreceptor-specific proteins, such as recoverin or arrestin, to metabolic proteins, such as enolase or carbonic anhydrase II. Surprisingly, the antigenic proteins mostly represent intracellular proteins involved in important cellular functions ranging from phototransduction to glycolysis.8 Different antiretinal AAbs frequently coexist in a single patient, creating antibody arrays related to the syndrome.3 However, not all AAb target antigens have been identified.
In our laboratory, we detected AAbs in patients with symptoms of CAR and AR that bound to a 23-kDa protein on the blot, but subsequently did not react with a purified recoverin. We hypothesized that there are other retinal proteins of a similar molecular weight that can be targeted by autoimmunity in retinopathy. Indeed, a proteomic analysis of the gel with separated human retinal proteins revealed a few potential 23-kDa protein candidates, including guanylyl cyclase-activating protein 1 (GCAP 1), heat shock protein 27 (HSP27), a series of Ras-related proteins Rab GTPases, glutathione S-transferase P, ATP synthase O subunit, mitochondrial precursor, and others. Our studies focused on patients who had AAbs against four 23-kDa-autoantigens that included HSP27, Rab6A, GCAP1, and GCAP2 as possible targets in CAR/AR.
We retrospectively analyzed CAR and AR patients' sera from the Oregon Health and Science University (OHSU) Serum Repository for the presence of AAbs against 23-kDa proteins. We evaluated 640 CAR patients and 2315 AR patients between 2005 and 2014, and identified 173/2955 (6%) individuals (107 women and 66 men, ratio F/M = 2.6; average age, 59 years) whose serum AAbs reacted with a retinal protein of approximately 23-kDa molecular mass on the Western blot with proteins extracted from human retina. However, 105 serum AAbs did not react with purified recoverin in subsequent assays. Demographics of patients with anti-23-kDa AAbs are presented in Table 1. The study was performed according to the guidelines of the Declaration of Helsinki. The OHSU Institutional Review Board approved the project. On average, the patients already diagnosed with cancer were 9 years older than patients without cancer identified at the time of testing.
Sera from patients were examined for reactivity with retinal proteins by Western blot analysis using proteins extracted from human retinas as described previously.7 After initial screening, a confirmation of antiretinal specificity was performed with purified recombinant human proteins as follows: recoverin (purified in the Adamus Laboratory), HSP27 (Abcam, Cambridge, United Kingdom), Rab6A (Sino Biological, Inc., North Wales, PA, USA), as well as GCAP1 and GCAP2 (expressed from pET11d vector in a BLRDE3pLysS Escherichia coli strain and purified using previously described procedures14,15). A sample of 0.25 μg protein was loaded per lane of 16% Bio-Rad Criterion XT Bis-Tris gel (Bio-Rad Laboratories, Hercules, CA). After electrophoresis in Tris/glycine buffer, the proteins were transferred to Immobilon membrane (Millipore, Billerica, MA) using a semidry apparatus. Then, individual strips were incubated with patient serum that was initially identified as having anti-23-kDa AAbs. The following control primary antibodies were used: rabbit anti-recoverin R2 (1:50,000; developed in the Adamus Laboratory), rabbit anti-GCAP1 and GCAP216,17 (1:10,000), rabbit anti-rab6A (1:2000; Abcam), mouse anti-HSP27 (1:2000; Thermo Fisher Scientific, Waltham, MA). The secondary antibody, goat anti-human IgG (H+L chain; Thermo Fisher Scientific), goat anti-rabbit IgG (H+L chain; Invitrogen), and goat anti-mouse IgG (H+L chain; Invitrogen), all conjugated to alkaline phosphatase were diluted 1:2000.
Human retinal cryosections (12 μm) in optimal cutting temperature (OCT) compound were postfixed with 4% paraformaldehyde for 10 minutes followed by blocking with 10% normal goat serum with 1% bovine serum albumin and 0.2% Tween in phosphate buffered saline (PBS) for 60 minutes at room temperature (RT). Then, human sera (diluted 1:50), each specific to one of 23-kDa protein as revealed by Western blotting analysis, were added and incubated overnight at 4°C. The next day, after washing with PBS anti-human IgG conjugated to Alexa Fluor 488 (1:1000; Invitrogen, Carlsbad, CA) was added for 1 hour of incubation. Then, the sections were washed and incubated with various specific primary antibodies for 1 hour at RT as follows: rabbit anti-bovine recoverin R2 that cross-reacted with human recoverin (diluted 1:500), anti-human Rab6A, anti-human HSP27, and anti-bovine CGAP116 that cross-reacted with human GCAP1 (diluted 1:200). After washing, the appropriate fluorescent secondary antibodies conjugated to Alexa Fluor 594 (1:2000, Invitrogen) were added for an additional 1 hour of incubation. The sections were washed in PBS and a mounting reagent containing 4′,6-diamidino-2-phenylindole (DAPI) was added to seal the sections, inhibit fluorescence quenching, and stain the nuclei. The immunofluorescent images were acquired using an Olympus Fluoview1000 confocal microscope, and pseudocolors were applied for analysis by Olympus FluoView FV10-ASW software (Olympus, Center Valley, PA). A negative control contained secondary antibodies only.
The identification of reactive 23-kDa molecular mass protein antigens was performed by Dr. Larry David in the OHSU Proteomics Shared Resource facility as described previously.18 Briefly, 30 μg portions of human retinal proteins were separated in 3 lanes of an SDS-PAGE using a Bio-Rad Criterion 10% gel stained with Coomassie brilliant blue and 2-mm wide slices excised from the bottom of the gel. The excised gel slices were destained twice for 30 minutes by shaking in 50 mM ammonium bicarbonate buffer, 50% acetonitrile, dried, and then reduced by addition of 10 mM dithiothreitol, 100 mM ammonium bicarbonate, and incubation at 56°C for 30 minutes. Proteins then were alkylated by addition of 55 mM iodoacetamide, 100 mM ammonium bicarbonate, and incubation at RT for 30 minutes in the dark. Gel slices then were again washed with destain solution, dried, and reswelled on ice for 15 minutes in digestion buffer containing 100 mM ammonium bicarbonate, 5 mM CaCl2, and 25 ng/μL sequencing grade modified trypsin (ProMega, Madison, WI). Following reswelling, excess trypsin solution was removed, gel slices covered with digestion buffer without trypsin, and samples incubated overnight at 37°C with shaking. Peptides then were recovered by addition of 3 μL of formic acid, shaking for 15 minutes, and the digestion buffer removed from the gel slices and placed in autosamper vials. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) of peptides was performed by capillary reverse phase separation using a 95-minute acetonitrile gradient and an LTQ linear ion trap mass spectrometer (Thermo Fisher Scientific) to generate data-dependent MS/MS. The resulting LC-MS/MS data then were analyzed to identify peptides and proteins using the Protein Discoverer 1.4 Suite (Thermo Fisher Scientific) with Sequest HT and Percolator programs to identify peptides and calculate peptide false discovery rates, respectively. Searches used a decoy database strategy and a human-only version of the Swiss-Prot database (Swiss Institute of Bioinformatics, Geneva, Switzerland) containing 20,163 entries and downloaded in July 2015. Only peptides with Percolator q scores below 0.01 (approximate false discovery rate <1%) and proteins with 2 or more unique peptide identifications in each gel slice were accepted. Protein Discoverer .msf files then were loaded into Scaffold 4.2.1 (Proteome Software, Portland, OR, USA) and numbers of MS/MS spectra assigned to each protein (spectral counts) in each gel slice exported. Under these criteria, a total of 1033 human retinal proteins were identified with 19 matches to decoy database entries, resulting in a protein false discovery rate of approximately 2%. The identified reactive 23-kDa molecular weight protein antigens investigated in this study are shown in Supplementary Figure S1.
Statistical analysis was performed with GraphPad Prism software (GraphPad Software, Inc., San Diego, CA, USA) using the Fisher's exact test, χ2 test, and 2-tailed Mann-Whitney U test. Differences with a P value <0.05 were considered significant.
All identified proteins of the apparent 23-kDa molecular weight migrated in the SDS-PAGE gel similarly to recoverin and could be taken as recoverin without following confirmation with a purified protein or subsequent sequencing. We identified 173 patients with anti-23-kDa AAbs, including 68 (39%) individuals having anti-recoverin AAbs. However, the remaining 105 sera had not reacted with purified recoverin. Hence, our goal was to identify the reactive 23-kDa retinal protein(s) for those patients' sera.
Representative blots showing immune reactivity with purified Rab6A, GCAP1/2, and HSP27 proteins. Individual blots containing purified proteins were incubated with patient serum diluted 1:30 or specific antibodies, marked here as "(+)," producing positive reaction. Arrows point at positive bands. In the case of GCAP proteins, patient's serum was incubated with a blot containing GCAP1 and GCAP2; it reacted only with one protein (marked as "+").
To further characterize these serum AAbs, we performed the confocal immunostaining studies of human retinas with representative only monospecific human sera, containing AAbs against 23-kDa proteins. The monospecific sera within the group (recoverin, HSP27, Rab6A, and GCAP1) showed a similar pattern of staining (n = 4).
As is presented in Figures 2A and 2B, anti-recoverin AAbs showed typical immunostaining of the whole photoreceptors cells (red color) and some bipolar cells. Double labeling studies showed overlapped staining of photoreceptor cells (yellow color).
Representative double immunofluorescent labeling images of outer human retina incubated with monospecific serum against Recoverin (A, B) and GCAP1 (C, D). (A–D) Blue color represents DAPI-stained nuclei, green color represents Alexa Fluor 488-conjugated anti-human antibodies staining of photoreceptor cell layer recognized by the CAR patient AAbs; red color represents Alexa Fluor 594-conjugated antibodies staining of the photoreceptor cell layer by specific antibodies; the fourth image represents overlapping images—yellow color indicates common labeling. (B-D) Magnified overlapped images. An arrow in (A) points at photoreceptor cells and in (B) at outer segments; an arrow on (C) points at outer segments of photoreceptors and in (D) shows labeling of the cone cell, the green arrow shows tips of photoreceptors that were labeled by the patient serum (green color). Boxed areas indicated the enlarged area in (B) and (D). ONL, outer nuclear layer; INL, inner nuclear layer.
The other anti-23-kDa sera showed a different pattern of staining, which further confirmed that those AAbs did not recognized recoverin in the tissue but some other antigen(s). Immunolabeling with anti-GCAP1–specific antibodies (red color) or patient's serum (green color) showed stained rod and cone outer segments, synaptic terminals, and some cone somata (Figs. 2C, 2D). The colocalization experiments with a patient serum and anti-GCAP1 revealed a positive signal in cone photoreceptor cells (yellow color). Anti-GCAP2 serum was not used because it was represented by only 1 patient.
Antibodies against Rab6A (red color) showed a strong and uniform immunolabeling in human retina localized to the inner plexiform layer and cytoplasm of the ganglion cells (Figs. 3A, 3B). Also, punctate staining of the inner limiting membrane was present. Photoreceptor cells were not intensely labeled, although some immunofluorescent staining was observed in the inner segments (not shown). Colocalization studies showed that patient serum (green color) immunolabeled the same cellular structures of ganglion cells as did anti-Rab6A (yellow color) in human retina, confirming that AAbs recognize the same antigenic protein.
Representative double immunofluorescent labeling images of inner human retina incubated with monospecific serum against Rab6A (A, B) and HSP27 (C, D). (A–D) Blue color represents DAPI stained nuclei, green color represents Alexa Fluor 488-conjugated anti-human antibodies staining with the CAR patient AAbs; red color represents Alexa Fluor 594-conjugated antibodies staining with specific antibodies; the fourth image represents overlapping images—yellow color indicates common labeling. An arrow in (A) points at the ganglion cells and in (B) at the punctuated staining of the nerve fiber layer; a star in (B) shows intense staining of the ganglion cells stroma; an arrow in (C) points at the ganglion cells and in (D) shows intense staining of the nerve fiber layer, the star shows the ganglion cell body labeled yellow; Boxed areas indicated the enlarged area in (B) and (D). (B, C) magnified overlapped images. IPL, inner plexiform layer; GCL, ganglion cell layer; NFL, nerve fiber layer.
Autoantibodies specific to HSP27 also immunolabeled the ganglion cell layer and nerve fiber layer in a human retina (red color). The representative confocal image of double stained retinas (Figs. 3C, 3D) using specific anti-HSP27 antibody and a human patient serum both produced a strong immunofluorescence in ganglion cells and nerve fibers, indicating similarity in immunolabeling of the same antigen (yellow color) in the retina.
Altogether, our findings revealed a discovery of the 4 novel CAR and AR autoantibody targets of similar molecular mass proteins, which we identified as a Rab6A, HSP27, GCAP1, and GCAP2. Sera specific for the above proteins showed a different immunostaining pattern relevant to their cellular location.
In total, 62 of 173 (36%) patients had been diagnosed with various kinds of cancers, including 3 bladder, 10 breast, 6 colon, 10 gynecologic, 13 lung, 2 renal, 3 prostate, 5 melanomas, and other cancers. In 14 seropositive patients with anti-23-kDa AAbs, the detection of serum AAbs preceded a diagnosis of cancer. Of 62 patients with cancer, 34 had anti-recoverin AAbs (55%), 19/62 (31%) had anti-Rab6A AAbs, and 8/62 (13%) had anti-HSP27 AAbs. Patients with anti-GCAP1/2 AAbs did not have cancer at the time of testing. The statistical analysis (Fig. 4) shows that patients with anti-recoverin AAbs had a significantly higher likelihood of having cancer than those with other anti-23-kDa AAbs (P = 0.0002, Fisher's exact test).
Statistical analysis of seropositive patients with and without cancer having anti-recoverin and other anti-23-kDa AAbs. (A) Four groups of seropositive patients with and without cancer were analyzed by χ2 analysis (***P = 0.0003, χ2 test) and t Fisher's exact test (significance denoted by asterisks). (B) Patients with anti-recoverin antibodies had higher likelihood of having cancer than with other anti-23-kDa AAbs (***P = 0.0002, Fisher's exact test). CA(+), patients with cancer; CA(−), patients without cancer.
Among our 173 patient cohort, 68 individuals (39%) were identified to be seropositive for anti-recoverin AAbs. Only 34 those 68 (50%) patients were reported to be diagnosed with cancer at the time of testing (Table 1). Patients with anti-recoverin AAbs and symptoms of CAR were older than the seropositive patients who had not been diagnosed with cancer by an average of 11 years. Seropositivity to recoverin was related to a rapid sudden onset of visual symptoms, and a progressive course. Table 2 compares the retinal phenotypes associated with anti-23–kDa AAbs, divided into 4 groups according to their antibody specificity and provides the frequencies of symptoms. Some patients had difficulty seeing in the dark (nyctalopia) and photopsia, and also peripheral vision loss, which may reflect the rod dysfunction more so than cone defects. One third of patients presented with reduced visual acuity from 20/70 to hand motion vision (HM), defects in visual fields, and severely impaired retinal function to unresponsiveness as determined by electroretinography (ERG), indicating loss of rod and/or cone function. Attenuated vessels, retinal atrophy, and vitritis were found in recoverin-seropositive patients.
The occurrence of anti-GCAP AAbs was rare. Autoantibodies binding to human GCAP1 were found only in 6 of 173 patients (3%) and 1 patient showed a reaction with GCAP2 on Western blots. The average age of anti-GCAP seropositive patients was 57 years, with an equal number of men and women. Of the GCAP1-seropositive patients, 67% reported loss of visual acuity and central vision loss, and 50% presented with maculopathy and cone dysfunction confirmed by ERG. Loss of vision was progressive. No association with cancer was reported; however, the small number of patients with such AAbs prevents us from drawing definitive conclusions.
Of the patients, 31% had AAbs against a newly identified target antigen Rab6A. Over 35% of those patients presented with cancer, including lung (4 patients), colon (3), breast (2), renal (2), lymphomas (2), and others. The average latency time between cancer, ocular symptom manifestation, and detection of circulating AAbs was 2.7 years, with 1 exception of a patient whose serum antibody preceded diagnosis of lung cancer by 3 years. The patients complained about progressive vision loss with defects in visual acuity ranging from 20/80 to HM. Visual field defect (34%), central and peripheral loss (11%), and loss of rod and cone function (23%) was documented. Retinal dystrophy and outer retinal thinning was reported in 43% patients.
Autoantibodiess against HSP27 have been reported previously in patients with glaucoma,20 and animal experiments revealed their pathogenic potential.21 We identified 45 (26%) sera that did not react with recoverin but bound strongly to human HSP27 on the blot (Fig. 1). Seropositive patients presented with slowly progressive central vision loss (20%), photophobia, photopsia, and nyctalopia, constriction of visual fields (central scotoma, enlarged blind spot) along with generalized loss of rod and cone function as confirmed by the ERG (36%). Optic nerve defect was not a predominant feature in this group, although 3 patients had been diagnosed with low tension glaucoma. Some patients presented with intraocular inflammation (18%). Associated infectious and autoimmune diseases (lupus) were reported in 8 (18%) patients. The majority of persons (37/45, 82%) in this group were without cancer at the time of testing.
Table 2 summarizes the clinical phenotypes of the 173 patients with anti–23-kDa protein AAbs based on seropositivity, symptoms, and findings using the following criteria: visual manifestation, clinical findings, ERGs, and presence of cancer. The clinical data were obtained from the summary information submitted with the specimen for antibody testing. Seropositivity to Rab6A, HSP27, and GCAP1 was compared with anti-recoverin phenotypes. The important difference compared to the prior findings is the incidence of cancer in patients with anti-recoverin AAbs. Until now, we and others reported almost 100% association of anti-recoverin AAbs with cancer. However, the current study identified only 50% recoverin-seropositive patients with cancer at the time of testing.
Our studies showed, for the first time to our knowledge, new autoantigens in CAR and AR. Among the cohort of 173 patients seropositive for 23-kDa protein, only 68 had reacted with recoverin and the remaining 105 had 4 AAbs targeting novel antigens of similar molecular mass, which we identified as a Rab6A, HSP27, GCAP1, and GCAP2. This discovery has an important implication for anti-recoverin antibody testing, and we always recommend a verification of antibody reactivity with purified recoverin protein before calling it recoverin-positive. Confocal images confirmed that these autoantigens were expressed in different types of cells in the retina, consistent with their respective cellular functions.
Retinal guanylyl cyclase (RetGC)-activating proteins (GCAPs) are calcium-sensitive proteins that have an essential physiologic role in photoreceptors by accelerating the recovery of rods and cones following excitation by light28 and they can trigger congenital retinal diseases in humans.29–31 Several mutations in the GCAP1 gene have been associated with autosomal dominant cone dystrophy in human patients. The loss of GCAP1 alters the recovery kinetics in GCAP1−/− mice but does not initiate a degenerative disorder.32 Therefore, the presence of the GCAP1 AAbs may more likely contribute to photoreceptor apoptosis rather than affecting normal photoreceptor function at the signaling level. Three of our patients seropositive for GCAP1 presented with cone dystrophy although their genetic status presently is unknown. So far, we identified only a single patient with anti-GCAP2 AAbs so we cannot presume their role in retinal disease.
Autoantibodies against Rab6A have not been reported before in disease. Rab proteins are small (21–25 kDa) monomeric GTPases/GTP-binding proteins, and form the largest branch of the Ras superfamily.33,34 There are at least 70 Rab genes in the human genome, and a number of Rab GTPases are conserved from yeast to humans.34–37 The different Rab GTPases are localized to the cytosolic surface of intracellular membranes, where they function as regulators of distinct steps in membrane traffic pathways.38 Rab1 and Rab6 GTPases have been found in the outer retina and likely have an important role in maintaining the structure and function of photoreceptors.39 In photoreceptors, rhodopsin is believed to be transported via a Rab6 regulated pathway and, thus, defects in trafficking pathways may lead to retinal degeneration.40 Our study also showed Rab6A to be present in the outer segment, and also in bipolar and the cytoplasm of the ganglion cells. Its role in autoimmunity has not been studied previously to our knowledge, but AAbs could potentially block those important cellular functions.
We have reported on AAbs specific for HSPs, including HSP60 that was a frequently recognized autoantigen by AAbs of patients with AR, irrespective of cancer status.4 In the current study, we present findings of AAbs against a small heat shock protein, HSP27, mostly in patients with AR and in 18% CAR patients diagnosed with different kinds of cancer. HSP27 is a protein that can act as a protective and harmful factor in pathologies, such as neurodegenerative diseases, myopathies, asthma, cataracts, and cancers.41,42 HSP27 has powerful neuroprotective effects, increasing the survival of cells subjected to cytotoxic stimuli.42 This is especially relevant to the study of the retina where cells die as a result of retinal disease and injury. The upregulation of this protein occurs in response to ischemia and oxidative stress, traumatic nerve injury, and elevated intraocular pressure and glaucoma.43 We believe that AAbs act as a stress to photoreceptor cells, and multiple antibodies may promote antibody-mediated retinal degeneration by blocking their functions.
In conclusion, testing for AAbs against retinal antigens provides important information for molecular diagnosis. The identification of over 1000 human retinal proteins classified by their migration by SDS-PAGE will help support identification of new retinal autoantigens following detection by Western blotting. The relevance of newly identified autoantigens (Rab6A, GCAP1, and HSP27) as biomarkers to diagnose, monitor disease activity/severity, and predict response to therapy is limited but they are promising candidates. These AAbs are associated with distinctive clinical subsets with different prognostic features. The mechanism of anti-Rab6A, anti-GCAP1, and anti-HSP27 autoantibody formation in patients with and without cancer is not fully understood, although immunity to recoverin and HSP27 has been shown to be pathogenic in animals. The fact that these AAbs are associated with symptoms of CAR and AR suggests their pathogenic function alone or in combination with other pathogenic anti-retinal AAbs. Circulating AAbs consistently precede the clinical onset of the autoimmune diseases by years, therefore, the benefits of anti-retinal AAbs testing include aiding with diagnosis of unique retinal presentation, in monitoring and prognosis of vison loss, treatment options, and also helping with early diagnosis of cancer. Whether AAbs are causative by reducing/extinguishing cellular functions (e.g., protective function of HSP27), leading to cell death, or are generated later as an effect of retinal degeneration, they also may enhance progression of undergoing retinal disease. An important observation in this large group of CAR and AR patients is that the association of anti-recoverin AAbs with the presence or future development of cancer was substantially lower than in prior reports.
Supported by Grant EY11522 from the National Institutes of Health (NIH; Bethesda, MD, USA), and by unrestricted Casey Eye Institute funding from Research to Prevent Blindness (New York, NY, USA). Mass spectrometric analysis was performed by Dr. Larry David in the OHSU Proteomics Shared Resource with partial support from NIH Core Grants P30EY010572 and P30CA069533.
Weleber RG, Watzke RC, Shults WT, et al. Clinical and electrophysiologic characterization of paraneoplastic and autoimmune retinopathies associated with antienolase antibodies. Am J Ophthalmol. 2005; 139: 780–794.
Adamus G, Brown L, Weleber RG. Molecular biomarkers for autoimmune retinopathies: significance of anti-transducin-alpha autoantibodies. Exp Mol Pathol. 2009; 87: 195–203.
Adamus G, Choi D, Raghunath A, Schiffman J. Significance of anti-retinal autoantibodies in cancer-associated retinopathy with gynecological cancers. J Clin Exp Ophthalmol. 2013; 4: 307.
Adamus G, Bonnah R, Brown L, David L. Detection of autoantibodies against heat shock proteins and collapsin response mediator proteins in autoimmune retinopathy. BMC Ophthalmol. 2013; 13: 48.
Heckenlively J, Ferreyra H. Autoimmune retinopathy: a review and summary. Sem Immunopathol. 2008; 30: 127–134.
Kondo M, Sanuki R, Ueno S, et al. Identification of autoantibodies against TRPM1 in patients with paraneoplastic retinopathy associated with ON bipolar cell dysfunction. PLoS One. 2011; 6: e19911.
Adamus G, Ren G, Weleber RG. Autoantibodies against retinal proteins in paraneoplastic and autoimmune retinopathy. BMC. 2004; 4: 5.
Adamus G. Autoantibody targets and their cancer relationship in the pathogenicity of paraneoplastic retinopathy. Autoimmun Rev. 2009; 8: 410–414.
Adamus G, Guy J, Schmied JL, Arendt A, Hargrave PA. Role of anti-recoverin autoantibodies in cancer-associated retinopathy. Invest Ophthalmol Vis Sci. 1993; 34: 2626–2633.
Polans AS, Buczylko J, Crabb J, Palczewski K. A photoreceptor calcium binding protein is recognized by autoantibodies obtained from patients with cancer-associated retinopathy. J Cell Biol. 1991; 112: 981–989.
Ray S, Zazula S, Niemi GA, et al. Cloning, expression, and crystallization of recoverin, a calcium sensor in vision. Proc Natl Acad Sci USA. 1992; 89: 5705–5709.
Adamus G. Paraneoplastic retinal degeneration. In: Levin L, Albert DM, eds. Ocular Disease: Mechanisms and Management. Atlanta, GA: Saunders Elsevier, Inc.; 2010: 599–608.
Rahimy E, Sarraf D. Paraneoplastic and non-paraneoplastic retinopathy and optic neuropathy: evaluation and management. Surv Ophthalmol, 2013; 58: 430–458.
Olshevskaya EV, Hughes RE, Hurley JB, Dizhoor AM. Calcium binding, but not a calcium-myristoyl switch, controls the ability of guanylyl cyclase-activating protein GCAP-2 to regulate photoreceptor guanylyl cyclase. J Biol Chem, 1997; 272: 14327–14333.
Peshenko IV, Dizhoor AM. Activation and inhibition of photoreceptor guanylyl cyclase by guanylyl cyclase activating protein 1 (GCAP-1): the functional role of Mg2+/Ca2+ exchange in EF-hand domains. J Biol Chem, 2007; 282: 21645–21652.
Olshevskaya EV, Calvert PD, Woodruff ML, et al. The Y99C mutation in guanylyl cyclase-activating protein 1 increases intracellular Ca2+ and causes photoreceptor degeneration in transgenic mice. J Neurosci. 2004; 24: 6078–6085.
Makino CL, Peshenko IV, Wen XH, Olshevskaya EV, Barrett R, Dizhoor AM. A role for GCAP2 in regulating the photoresponse. Guanylyl cyclase activation and rod electrophysiology in GUCA1B knock-out mice. J Biol Chem. 2008; 283: 29135–29143.
Spinelli KJ, Klimek JE, Wilmarth PA, et al. Distinct energy metabolism of auditory and vestibular sensory epithelia revealed by quantitative mass spectrometry using MS2 intensity. Proc Nat Acad Sci USA. 2012; 109: E268–E277.
Polans AS, Crabb JW, Palczewski K. Calcium Binding Proteins in the Retina. San Diego, CA: Academic Press, Inc.; 1993: 248–260.
Wax MB, Tezel G, Kawase K, Kitazawa Y. Serum autoantibodies to heat shock proteins in glaucoma patients from Japan and the United States. Ophthalmology. 2001; 108: 296–302.
Wax MB, Tezel G, Yang J, et al. Induced autoimmunity to heat shock proteins elicits glaucomatous loss of retinal ganglion cell neurons via activated T-cell-derived Fas-ligand. J Neurosci. 2008; 28: 12085–12096.
Senin II, Koch KW, Akhtar M, Philippov PP. Ca2+-dependent control of rhodopsin phosphorylation: recoverin and rhodopsin kinase. Adv Exp Med Biol. 2002; 514: 69–99.
Adamus G, Ortega H, Witkowska D, Polans A. Recoverin: a potent uveitogen for the induction of photoreceptor degeneration in Lewis rats. Exp Eye Res. 1994; 59: 447–456.
Adamus G, Machnicki M, Seigel GM. Apoptotic retinal cell death induced by autoantibodies of cancer associated retinopathy. Invest Ophthalmol Vis Sci. 1997; 38: 283–291.
Adamus G, Machnicki M, Elerding H, Sugden B, Blocker YS, Fox DA. Antibodies to recoverin induce apoptosis of photoreceptor and bipolar cells in vivo. J Autoimmun. 1998; 11: 523–533.
Polans A, Witkowska D, Haley T, Amundson D, Baizer L, Adamus G. Recoverin, a photoreceptor-specific calcium-binding protein, is expressed by the tumor of a patient with cancer-associated retinopathy. Proc Natl Acad Sci USA. 1995; 92: 9176–9180.
Miyagawa Y, Ohguro H, Odagiri H, et al. Aberrantly expressed recoverin is functionally associated with G-protein-coupled receptor kinases in cancer cell lines. Biochem Biophys Res Commun. 2003; 300: 669–673.
Pennesi ME, Howes KA, Baehr W, Wu SM. Guanylate cyclase-activating protein (GCAP) 1 rescues cone recovery kinetics in GCAP1/GCAP2 knockout mice. Proc Natl Acad Sci USA. 2003; 100: 6783–6788.
Olshevskaya EV, Ermilov AN, Dizhoor AM. Factors that affect regulation of cGMP synthesis in vertebrate photoreceptors and their genetic link to human retinal degeneration. Mol Cell Biochem. 2002; 230: 139–147.
Dizhoor AM, Woodruff ML, Olshevskaya EV, et al. Night blindness and the mechanism of constitutive signaling of mutant G90D rhodopsin. J Neurosci. 2008; 28: 11662–11672.
Woodruff ML, Olshevskaya EV, Savchenko AB, et al. Constitutive excitation by Gly90Asp rhodopsin rescues rods from degeneration caused by elevated production of cGMP in the dark. J Neurosci. 2007; 27: 8805–8815.
Makino CL, Wen XH, Olshevskaya EV, Peshenko IV, Savchenko AB, Dizhoor AM. Enzymatic relay mechanism stimulates cyclic GMP synthesis in rod photoresponse: biochemical and physiological study in guanylyl cyclase activating protein 1 knockout mice. PLoS One. 2012; 7: e47637.
Pereira-Leal JB, Seabra MC. The mammalian Rab family of small GTPases: definition of family and subfamily sequence motifs suggests a mechanism for functional specificity in the Ras superfamily. J Mol Biol. 2000; 301: 1077–1087.
Bhuin T, Roy JK. Rab proteins: the key regulators of intracellular vesicle transport. Exp Cell Res. 2014; 328: 1–19.
Darchen F, Goud B. Multiple aspects of Rab protein action in the secretory pathway: focus on Rab3 and Rab6. Biochimie. 2000; 82: 375–384.
Stenmark H, Olkkonen VM. The Rab GTPase family. Genome Biol 2001; 2:REVIEWS 3007.1–REVIEWS3007.7.
Tong C, Ohyama T, Tien AC, Rajan A, Haueter CM, Bellen HJ. Rich regulates target specificity of photoreceptor cells and N-cadherin trafficking in the Drosophila visual system via Rab6. Neuron. 2011; 71: 447–459.
Cheng KW, Lahad JP, Gray JW, Mills GB. Emerging role of RAB GTPases in cancer and human disease. Cancer Res. 2005; 65: 2516–2519.
Huang W, Wu G, Wang GY. Cell type-specific and light-dependent expression of Rab1 and Rab6 GTPases in mammalian retinas. Vis Neurosci. 2009; 26: 443–452.
Shetty KM, Kurada P, O'Tousa JE. Rab6 regulation of rhodopsin transport in Drosophila. J Biol Chem. 1998; 273: 20425–20430.
Arrigo A-P, Simon S, Gibert B, et al. Hsp27 (HspB1) and [alpha]B-crystallin (HspB5) as therapeutic targets. FEBS Lett. 2007; 581: 3665–3674.
Franklin TB, Krueger-Naug AM, Clarke DB, Arrigo A-P, Currie RW. The role of heat shock proteins Hsp70 and Hsp27 in cellular protection of the central nervous system. Int J Hyperthermia. 2005; 21: 379–392.
O'Reilly AM, Currie RW, Clarke DB. HspB1 (Hsp 27) expression and neuroprotection in the retina. Mol Neurobiol. 2010; 42: 124–132.
Tezel GM, Seigel GM, Wax MB. Density-dependent resistance to apoptosis in retinal cells. Curr Eye Res. 1999; 19: 377–388.
Garrido C, Gurbuxani S, Ravagnan L, Kroemer G. Heat shock proteins: endogenous modulators of apoptotic cell death. Biochem Biophys Res Commun. 2001; 286: 433–442.
Kalesnykas G, Niittykoski M, Rantala J, et al. The expression of heat shock protein 27 in retinal ganglion and glial cells in a rat glaucoma model. Neuroscience. 2007; 150: 692–704.
Tezel G, Wax MB. The mechanisms of hsp27 antibody-mediated apoptosis in retinal neuronal cells. J Neurosci. 2000; 20: 3552–3562.
Laspas P, Gramlich OW, Müller HD, et al. Autoreactive antibodies and loss of retinal ganglion cells in rats induced by immunization with ocular antigens. Invest Ophthalmol Vis Sci. 2011; 52: 8835–8848.
Tezel G, Seigel GM, Wax MB. Autoantibodies to small heat shock proteins in glaucoma. Invest Ophthalmol Vis Sci. 1998; 39: 2277–2287.
Gramlich OW, Bell K, von Thun UndHohenstein-Blaul N, et al. Autoimmune biomarkers in glaucoma patients. Curr Opin Pharmacol. 2013; 13: 90–97.
Olejek A, Damasiewicz-Bodzek A, Bodzek P, et al. Concentrations of antibodies against heat shock protein 27 in the sera of women with ovarian carcinoma. Int J Gynecol Cancer. 2009; 19: 1516–1520.
Lianos GD, Alexiou GA, Mangano A, et al. The role of heat shock proteins in cancer. Cancer Lett. 2015; 360: 114–118.
Korneeva I, Bongiovanni AM, Girotra M, Caputo TA, Witkin SS. Serum antibodies to the 27-kd heat shock protein in women with gynecologic cancers. Am J Obstet Gynecol. 2000; 183: 18–21.
Chen P, Shi L, Jiang Y, et al. Identification of heat shock protein 27 as a novel autoantigen of Behcet's disease. Biochem Biophys Res Commun. 2015; 456: 866–871.
Pourghadamyari H, Moohebati M, Parizadeh SM, et al. Serum antibody titers against heat shock protein 27 are associated with the severity of coronary artery disease. Cell Stress Chaperones. 2011; 16: 309–316.


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text:
Public Inquiry announced into clinical practice of urology consultant
Tuesday, November 24th, 2020 11:22am
By Grainne McKinney @grainnemckinney
Health Minister Robin Swann has ordered a statutory public inquiry in relation to serious concerns about the clinical practice of a hospital consultant.
The decision, announced in a statement to the Assembly, relates to the work of a urology consultant who earlier this year retired from the Southern Trust.
The Health Minister has also informed MLAs that the independent inquiry panel set up in relation to the recall of neurology patients at Belfast Trust will be converted to a statutory public inquiry.
This is to enable the inquiry panel team to complete its work with "unfettered access to all relevant information", he stated.
This incident concerns the clinical practice of a urology consultant, Mr Aidan O'Brien, who earlier this year retired from the Southern Trust.
On 31st July 2020 the Southern Trust contacted the Health Department to report an Early Alert concerning the clinical practice of this consultant.
On the 7th June 2020 it became aware of potential concerns regarding delays of treatment of surgery patients who were under the care of the consultant urologist employed by the Trust.
The Trust became aware that 2 out of 10 patients listed for surgery under the care of this consultant were not on the hospital's Patient Administration System at that time.
As a result of these potential patient safety concerns, an initial lookback exercise in relation to the consultant's work was conducted, to ascertain if there were other areas of potential concern.
This initial lookback, which considered cases over a 18 month period of the consultant's work in the Southern Trust (from 1st January 2019 – 30th June 2020), concentrated on whether patients had a stent inserted during a particular procedure and if this stent had been removed within the clinically recommended timeframe.
Concerns were identified with 46 cases within a total of 147 patients who had the particular procedure and were listed as being under the care of the consultant during the period addressed by the initial lookback exercise
Minister Swann told MLAs that to date 1,159 patients' records have initially been reviewed and 271 patients or families have been contacted by the Trust.
So far 9 cases have been identified that meet the threshold for a Serious Adverse Incident (SAI) review and all 9 patients and / or their families have been contacted by the Trust to inform them of the position in relation to their respective cases.
The consultant also had a significant amount of private practice and that much of this was carried out in private domestic premises, therefore sitting outside of the regulatory framework.
Minister Swann says this is "of significant concern" as many of these patients may be unknown to the Southern Trust or the wider HSC system.
When his Department was contacted on the 31st July 2020, the Trust confirmed the following actions had already been taken:
 Discussions with the GMC employer liaison service had been conducted;
 The case had been discussed with NHS Resolutions who recommended restrictions of clinical practice;
 Restrictions were put in place by the Trust preventing the consultant from undertaking clinical work within the Trust and denying him access to or the ability to process patient information;
 The Trust also requested that the consultant voluntarily undertake to refrain from seeing any private patients at their home or any other setting and that he would confirm the same in writing; and
 A preliminary discussion was also undertaken with the Royal College of Surgeons Invited Review Service regarding the consultant's practice and the potential scope and scale of any further lookback exercise which may be required.
Minister Swann said the concerns about clinical practice of the consultant urologist were "of the gravest concern":
"Unfortunately we are only at the start of what is likely to become a long and detailed investigation into this matter.
As Health Minister I want to firstly unreservedly apologise to these patients and their families for any upset and distress this has caused.
I would also wish to reassure them that I will endeavour to ensure that they obtain appropriate treatment, support and the care they need over the coming weeks and months."
The Trust has advised patients that if they are concerned about aspects of their Urology care and require further advice that they contact the Southern Trust by email or on its Urology Information Phone Line.
The information line is available Monday – Friday 10:00 - 15:00 and the number is 0800 4148520.

redpajama_set_name: RedPajamaCommonCrawl


text:
Q: Thread Not executing when WaitforMultipleObjects() function is used I am trying to make a test console application which opens and downloads data from different COM ports using different threads . After downloading data I am using WaitForMutipleObjects() to wait for download to complete from all the threads.
WaitForMultipleObjects(nThreadCount,m_threadHandle,true,INFINITE);

But this is not working , thread is returning without being executed . Here is my thread function.
void* Test::GetData(void *p)
{
   Test *pThis = (Test *)p;
   string strCOMport;
   int nChannelNo,nBaudRate;
   cout<< " Enter COM port "<<endl;
   cin>>strCOMport;
   cout<< " Enter Channel Number"<<endl;
   cin>>nChannelNo;
   cout<< " Enter Baud Rate "<<endl;
   cin>>nBaudRate;

   if(pThis->InitPort(nChannelNo,0x00,(unsigned char *)strCOMport.c_str(),nBaudRate,0x00) == 0x00)
       cout<< "Init port Success"<<endl;
   else
   {
       cout<< "failed";
       return NULL;
   }
   // download
   ExitThread(0);
}

These are the functions which are creating and Waiting for threads respectively
void Test::init()
{
  m_threadHandle[nThreadCount] = CreateThread(NULL,0,(LPTHREAD_START_ROUTINE)GetData,(LPVOID)this,0,&m_dwThreadID[nThreadCount]);
  nThreadCount++;
}

void Test::WaitForThreads()
{
  WaitForMultipleObjects(nThreadCount,m_threadHandle,true,INFINITE);
}

Why is the thread function returning abruptly?

A: Your thread procedure has the wrong signature. Note that you used a cast to make your code compile. Your code probably originally said:
CreateThread(..., GetData, ...);

And the compiler objected to GetData saying that it was not compatible with a parameter of type LPTHREAD_START_ROUTINE. Unfortunately you chose the wrong solution. Casting it to LPTHREAD_START_ROUTINE does not actually make GetData be a LPTHREAD_START_ROUTINE. It just shuts the compiler up.
CreateThread(..., (LPTHREAD_START_ROUTINE)GetData, ...);

Here, GetData is still not a LPTHREAD_START_ROUTINE, but now the compiler is not able to save you from yourself.
So, you need to declare GetData to have the correct signature.
DWORD WINAPI GetData(LPVOID lpParameter);

And this must be a non-member function, or a static member function.
Once you fix all this you will find that you no longer need to call ExitThread(). You can simply write return 0 from your thread function.

redpajama_set_name: RedPajamaStackExchange


text:
SEX OFFENDER REGISTERS WITH SHERIFF'S OFFICE
Jayson A. Sherer
The Sheriff's Office is notifying the public that a sex offender has moved into Baxter County from Wisconsin and has registered with the Sheriff's Office.
NAME: JAYSON A. SHERER
OLD ADDRESS: N5430 State Rd 76
Shiocton, Wisconsin 54170
NEW ADDRESS: 7108 Hwy 126 North #20
Midway, Arkansas 72651
The offender was convicted of sex crimes in the State of Wisconsin and ordered to register as a sex offender. He was arrested in December, 1991 in Wisconsin for the offense of Sexual Assault in the Second Degree and arrested in May, 2000, also in Wisconsin, for the offense of Second Degree Sexual Assault of a Child, which involved sexual contact with an underage female. The offender was also arrested for Failure to Register as a Sex Offender in Marion County, Arkansas in August, 1997.
(This offender has not yet been assigned a Risk Level by SOCNA (Sex Offender Community Notification Assessment).


text:
Binance Ideas
Best Crypto Ideas Platform
Binance Network – Building a Giant Pool of Free and Legitimate Assets
Binance, a leading online peer to peer lending marketplace has announced a new service called Binance Best Crypto Market. Cryptocurrency exchange startup Binance says it is launching a new market for digital art called non-fungal tokens or NFSs, an effort to tap the emerging interest in owning digital art tied to the distributed ledger technology of the future. Artists with "Premium Event" collections will get 90% of the profits from selling their work through Binance NFSs.
Binance plans to enable artists to sell their works using a new marketplace, which will be integrated with its own private marketplace that offers artists the ability to convert their collections into cryptosystems like Binance NFSs. Cryptocurrency marketplace makers like Binance are responding to growing demand for more secure ways to buy and sell digital works. Digital art lovers have long been concerned that traditional exchange routes like PayPal and credit cards were not sufficiently secure. These concerns have increased pressure on artists to build more secure relationships with buyers, many of whom may lack adequate funds to participate in traditional exchanges like PayPal. Binance, in response, has introduced an innovative new product that would allow artists working with the Binance platform to accept payment in the ether as well as credit card and Pay Pal using zero fees and free global gateways.
Binance is also introducing a new series of educational programs that would provide consumers with exposure to the ecosystem and how it works. The company recently announced the partnership with Acclaimed University to launch Binance Academy, a one-of-a-kind online learning initiative designed to help educate borrowers about the benefits of using a non-fungulent token exchange to purchase digital art. Binance Academy will feature top industry experts who will explain how Binance works, gain market intelligence, and share industry information. Acclaimed University is a non-profit educational network dedicated to helping students pursue a college degree. Binance Academy will be hosted at Acclaimed University's University Park campus.
Other noteworthy collaborations include participation in the World Future Society summit in Abu Dhabi, the first exchange between a major Cryptocurrency marketplace creator and a major Forex trading organization. The creators of Binance NFSs are slated to join forces with Oanda, one of the largest venture capital firms in the world, which is working on projects aimed to increase the liquidity and usability of the Binance NFSs. Other notable co-venturers include Triton Network and Teleflora, two of the largest trade networks in the world.
In addition, Binance is joining forces with several well-known organizations in the realm of venture capital. Among those joining forces are IDG Capital, accelerators including accelerator platform accelerator, Lightspeed Venture Capital, as well as Zen Ventures, a strategic investment firm started by two entrepreneurs. Binance also has formed strategic alliances with several well-known banking institutions including Bank of America, Wells Fargo Bank, Chase Bank, and Citibank. Binance expects to expand its list of corporate and private investors in the coming months, as it looks forward to leveraging its equity and growth capital.
Binance, the fastest growing venture capital marketplace in the world, has joined forces with a number of the biggest names in the realm of enterprise, media, technology, and finance to build the largest and most sophisticated collection of premium digital currencies ever assembled. Binance NFT, which is also referred to as Binance Network, is the brainchild of executives who previously built the Forex automaton, FAP Turbo. The developers behind the project, which is in its early stages, have now released Binance NFT for public consumption.
You can click here to become a member of the world's best trading platform, binance.
Binance, NFT, And The Future Of Regional Tastes
Binance Sign Up Review – What You Need to Know
A Brief Introduction to Binance
A Look At The Binance Exchange And Its Work Behind The Currencies It trades In
An Open Source Alternative to Traditionally Reliable Exchanges
Why Binance Is the Best Platform to Buybtc?
How To Sell Btc For Usd Using BINance


text:
Dance Deewane () is an Indian dance reality show that airs on Colors TV. The show features dance performers, including solo acts, Duo and groups representing any style of dance, competing for a grand prize.

The series premiered on Colors TV on 2 June 2018. The First season aired from 2 June 2018 to 15 September 2018, the second season aired from 15 June 2019 to 28 September 2019 and the third season aired from 27 February 2021 to 10 October 2021. Dance Deewane Juniors, the series' spin-off, aired from 23 April 2022 to 17 July 2022.

Synopsis
Various talented dancers are divided into three age groups, called "generations". The first generation is the youngest while the third generation is the oldest. In the end, the best-performing dancers from each generation compete against each other in the finale. The dancers usually perform alongside choreographers.

Details

Judges & Hosts

Details

Contestants

 Eliminated
 Quit
 Finalist
 1st Generation Winner
 2nd Generation Winner
 3rd Generation Winner
 Gang Pratik-Winner
 Gang Sonali-Winner
 Gang Tushar-Winner
 Ultimate Winner

References

External links

 Dance Deewane at IMDb

2018 Indian television series debuts
Colors TV original programming
Indian dance television shows
Indian reality television series
Dance competition television shows
Hindi-language television shows

redpajama_set_name: RedPajamaWikipedia


text:
Q: Добавление и удаление класса при нажатии на элемент У меня есть элемент label, мне надо что бы при нажатии на него, к нему добавлялся класс check. При повторном нажатии на элемент необходимо удалить данный класс.
Мне надо написать код на чистом JS.
Вот что я пытался сделать, но ничего не работает.
var checked = document.querySelector('#add_check');

checked.onclick = function(){
    this.classList.add('check')
};
checked.onclick = function(){
    this.classList.remove('check')
};

Второй вариант:
var checked = document.querySelector('#add_check');

checked.onclick = function(){
    this.classList.add('check')

    if(this.classList.contains('check')){
        this.classList.remove('check')
    }
};

`

A: *

*Свойство объекта может иметь только одно значение. Вторым присваиванием вы перетираете предыдущее.

*Вам нужно определиться, использовать toggle или ручную логику. В совокупности они друг другу мешают

document.querySelector('.label')
  .onclick = function() {
  if (this.classList.contains('checked')) {
  this.classList.remove('checked')
  } else {this.classList.add('checked')}
}
.label {
  background-color: yellow;
}

.label.checked {
  background-color: green;
}

Label

Если необходимо навешивать более одного обработчика, да и вообще для единства стиля, лучше использовать "слушатели событий" - event listeners

document.querySelectorAll('.label').forEach(label =>
  label.addEventListener('click', function() {
    this.classList.toggle('checked');
  }))
.label {
  background-color: yellow;
}

.label.checked {
  background-color: green;
}

Label

Label

A:

var checked = document.querySelector('#add_check');
checked.onclick = function(){
    this.classList.toggle('check');
    console.log(checked)
};

add_check


HoloData is a Virtual Reality Visualization tool that allows you to view Three Axis Graph data over time, letting you scroll through the time indices to see the 3d graph change before you, providing insights into the trends of your chosen data.
Extract the .zip file to your chosen directory.
Ensure your Oculus Rift and Leap-motion device are working correctly.
Enjoy the mental stimulation of seeing the 3d graph data!
At least! one App that works on Oculus Runtime 0.8!!!


Antique French Louis XVI style fireplace in limestone with marble inlay, mantel with authentic patina. Dimensions approx : overall width 156cm (61.5ins) x ht 110cm (43ins) x front-to-back 31cm (12ins). Width between side jambs 130cm (51 ins) x ht below lintel 89cm (35ins). Export document supplied.


text:
YouTube Ruined My Friendships
My very best friend hates social media and believes technology takes away the ability to enjoy the moment. I agree, but unplugged events won't contribute to my YouTube success as I must stay relevant on other platforms. Let's address how YouTube ruined my friendships.
Unless you live on an artsy college campus or in LA, you most likely aren't surrounded by fellow YouTubers, wannabe actors/actresses, and bored enough individuals with hefty trust funds. Some upcoming YouTubers may have 1-2 engaging friends (actively following your content and/or helping you produce) if they are lucky. But what if your friends don't understand or enjoy the entertainment industry altogether? You can experience several different emotions...
You Appear to be a User
Maybe a simple boomerang during your cheers at the table is a cute upload to your Instagram story. But then you gotta Snapchat your food. But what will be your official Instagram upload to document your outfit? Sure your friends may pull out their phone to shoot a text and take one selfie, but you find yourself constantly documenting and staging a social occasion that screams
I have friends!
You Seem Self-Absorbed
You make your friends pose and retake pictures to only ignore them. I mean, the pictures aren't going to filter themselves.
You do have a virtual audience to impress. You go out on week days and eat expensive meals. Flexing gets old for us influencers, but we are committed to this social game. If we dedicate too many minutes to the internet world, please speak up. We ultimately care about our friends.
Don't even get me started about vlogging. A lot of people have Instagram and Snapchat, but they cater to their inner circle instead of the general public. These friends have jobs and don't want to be filmed at a bar. They are shy and don't know what to say. Please give us three minutes to vlog the night and get exactly what we need.
You Feel Socially Awkward
You cringe to pull out your vlogging camera, but you must update your audience. Do you have to check with the people you're with to do so? What if you don't know them well? And wow, you don't feel like filming anymore.
But a plea to our friends... Talking to a camera in public feels really dorky. That's why you still rarely see vloggers when you go to the mall. It takes a lot of confidence to interact with "nobody" in front of everybody. Please don't roll your eyes and shame us for trying to build our brand.
Did I miss anything? If so, please sound off in the comments. If you're a small YouTuber struggling, please know that you can always reach out to me. YouTube is a wonderful place, but it can be it's own social bubble.
Want to read more in my 'Small YouTuber' series? Check out these articles:
How to Support YouTubers for FREE
YouTube Destroyed My Dating Life


There's nothing better than completing a fresh engine rebuild. When the wait for parts is over, the machining, assembly and wiring has been done and everything's been buttoned up tight. That's what we're looking at here. A freshly rebuilt 1975 Z1B motor that's ready to rumble. Let's walk you through the process.
While not the worst engine we've ever seen, we knew this diamond in the rough was going to require a little bit of polishing.
After breaking it the rest of the way down, a quick trip to the vapor honing cabinet got our parts back to pristine condition and we could move forward.
Heads are looking like new! We're going to be rebuilding them with stainless steel valves after resurfacing.
After being resurfaced and bored from 903cc to 1015cc we installed a Wiseco piston kit.
Here the heads are being reassembled using a new cam chain and guide, cam bearings top and bottom, chain dampeners and a new cam chain idler damper.
Here we're installing the rebuilt crank shaft using all new bearings and seals.
Here we're installing the Dyna S ignition before final assembly.
Now that final assembly is complete, we can check out this before and after shot.


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text:
August 30, 2017 by passage2truth
Texas Trilogy 1: Dreidel in Texas
This is a slightly re-edited and expanded version of a slideshow I did about five months ago, probably the second one I compiled this year (I did two in all of last year). I originally called it "Random Memories of East Texas," in part because I wanted a deliberately flat title to balance the rather sensational subject matter, but also beacause I realized it was an attempt–however flawed–to come to terms with living in East Texas for twenty years. It wasn't a very good or even very accurate title, as most of the events it focused on didn't even take place in East Texas, but in Dallas and Austin. Also, although they were someone's Texas memories, they weren't really my memories, since I didn't start living in Texas until 1991, well after the tragic events depicted in the first two movements of the slideshow. Also, I felt that James Bryd Jr.'s killing, which I was at least in Texas for (although over fifty miles away from Jasper), really didn't get adequate treatment, in part because the song ended too soon for my purposes. In the re-edited, expanded, and re-titled version, I have extended the audio track with some sound effects, thus allowing me to insert another six or seven slides in the final section.
I'm not sure it has a message in the conventional sense, beyond the obvious point that Texas can be a dangerous place, in part because of its culture, and in part because it's simply so big that some more or less random bad stuff is bound to happen. I always liked Don Maclean's song "Dreidel," and I have a fairly clear memory of him performing it on some daytime talk show about the time this album (his third) was released. I'm fairly sure it was a song (like the "The Pride Parade") that he wrote to explore his own mental situation in the wake of the massive success of his American Pie album that had come out the year before. Nevertheless, I think most people have a tendancy to personalize the songs they like and listen to a lot, and I'm sure I tended to think of both songs as in some sense reflections on my own feelings on entering high school in Southern California in the early seventies. I don't think that sense of identification ever fully left me, even when I stopped listening to Don Maclean (probably a mistake on my part, and doubtless one of many). The immediate stimulus for this slideshow was probably watching "The Tower" on Netflix, a brilliant documentary looking at the Austin clocktower murders of 1966, an incident I really don't remember (I would have been nine), although I remember my mother talking about it. Similarly, I was even younger when President Kennedy was assasinated, and I'm fairly sure I did not see it live (it seems unlikely it was even carried live on most national television stations, although it may well have been in the Dallas area). I do remember my mother hearing about it on the radio and talking with our next door neighbor, Mrs. Leddy, over the wall separating out two backyards from one another. I think they were crying, but I may be embellishing the memory (I would have just turned six a couple weeks before).
James Bryd Jr.s death was widely reported in Smith County where I was living at the time, and I am pretty sure many local people were aware that it made the national news, which tended to make the people I knew uncomfortable. I even remember the joke making the rounds at the time ("What red and black and two miles long"–I think you can guess the answer, even if you haven't heard it before). Humor is of course one way human beings tend to express and deal with discomfort, although it is also a way of expressing and reinforcing power relationships, often making sure that marginal groups stay marginalized. I suspect both were at work here, although I am sure some would disagree, especially as meanings of things like jokes tend to change according to time and context, so that one particular performance may well convey different meanings, and even different listeners may take different meanings away, some perhaps quite different than the teller consciously intended. Anyway, here is the first of three slideshows about Texas (the other two are both quite different, and were made several months after the first version of this one).
Posted in Culture, Existential Concerns, History, Personal, Politics, Slideshows, Texas | Tagged Charles Whitman, Clock Tower Sniper, Don Maclean, Dreidel, Hate Crimes, History, Jackie Kennedy, James Byrd Jr., Kennedy Assasination, Lee Harvey Oswald, Texas, UT Austin Killings | Leave a comment
Donovan's "Lalena": A unofficial slideshow about prostitutes
[Cross-posted on my Donovan–Slideshows by passage2trth page] Donovan's "Lalena" was a song I always admired without ever really thinking about it very much. Certainly, the song's most striking feature was Donovan's rather lovely vibrato and its melancholy mood, a mood I realize I have always found attractive. While of course I knew that it was about a girl with an unusual name who seemed rather sad, I never really thought about the implications of the lyrics until quite recently, when I was looking for a followup to "Universal Soldier." Once I did pause to think about who the lyrics were describing, it seemed rather obvious she was a prostitute, getting up "when the sun goes to bed," one whose "lot in life" makes the song's narrator sad even as he refuses to blame her for her situation, presumably because it is beyond her control. While by no means the only song ever written about what is frequently characterized as the world's oldest profession (cf. "Love for Sale, "Sweet Georgia Brown," "Roxanne," among many others), it is sort of surprising that "Lalena" was an AM hit in the sixties reaching number #33 on the Hot 100 in Billboard. Of course, so was The Animals version of "The House of the Rising Sun." In fact, Donovan was apparently inspired to write the song by Lotte Lenya's character of Jenny (a streetwalker) in the 1931 film version of Berthold Brecht and Kurt Weil's The Threepenny Opera. Donovan seems to have added an extra syllable to her surname in order to make the haunting chorus work musically, while the "lot in life" line is quite probably a gentle play the actress's first name.
Having decided to make the slideshow into a mini-history of prostitution, and wanting to be true to the song in both being not too graphic and rather melancholy, I chose to use Final Cut Pro (only the second time I've done so, and I'm still learning how) to do a series of overlays, often drawn from impressionist, post-impressionist, and modernist art (with a few photographs) in order to convey the song's–and the woman's–sadness at her situation. It may be a bit too busy, but I think it generally works in the way it gets across a deep underlying melancholy that is central to the song's appeal. The audio, by the way is not the single version, but a version Donovan performed for Italian public television in 1968.
Posted in Culture, Donovan, History, Slideshows | Tagged Donovan, History, Lalena, Lotte Lenya, Melancholy, Prostitution | Leave a comment


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Yorkshire voted top spot for UK holidays
Okt 9 2012 Veröffentlicht von Laura Canning
Yorkshire has been voted the top place to visit in the UK at the nineteenth World Travel Awards, voted for by travel agents from around the world.
The other nominees in the category were Scotland, Wales, the Peak District, Cornwall, Cumbria, Devon and Northumberland.
Yorkshire saw over 216 million visitors last year from all over the world, coming to see famous places such as the Yorkshire Dales National Park, the seaside town of Scarborough, Brontë country, and the historic cities of York and Whitby.
Statistics from January to September last year show there was a 12 per cent increase in visits by international tourists to the region.
Gary Verity, the chief executive of Welcome to Yorkshire, said: 'The win is testament to all the thousands of fantastic businesses we have in Yorkshire that time and again put the county on the map for millions of holidaymakers, not just in Britain, but the world over.'
Yorkshire is also vying to host the opening of the Tour de France in 2016, which starts in a foreign country every two years. 150,000 people have already backed the Yorkshire bid.
To see Yorkshire for yourself, we have hundreds of sites in North, South, East and West Yorkshire, from tent and tourer pitches to caravans for hire to lodges, with sites searchable by areas including Scarborough and Richmond, by price, by availability and by 80+ filters including sites which are open all year.


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Q: create a file structure for a 2D grid so for my A* algorithm I want to create a settings file that contains the data information for the loops creating the grid.
Currently my cells just need to know their field cost but this might change later on. I want to keep the information flexible.
A simple way would be creating a byte array

but I can only store the field cost in there. I thought about taking JSON for this
{
  "1": [ // my first row
    {
      "x": 1, // cell one in row one
      "cost": 1
    },
    {
      "x": 2,
      "cost": null
    },
    {
      "x": 3,
      "cost": 3
    },
    {
      "x": 4,
      "cost": null
    },
    {
      "x": 5,
      "cost": 2
    }
  ],
  "2": [ // my second row
    {
      "x": 1, // cell one in row two
      "cost": 3
    },
    {
      "x": 2,
      "cost": 2
    },
    {
      "x": 3,
      "cost": null
    },
    {
      "x": 4,
      "cost": 1
    },
    {
      "x": 5,
      "cost": 2
    }
  ]
}

By using JSON I can store more data to cells but this structure seems to be way more complex than a simple byte array.
Are there any better solutions for this?

A: Don't use JSON just read for a text file.
String input = File.ReadAllText( @"c:\myfile.txt" );

int i = 0, j = 0;
int[,] result = new int[10, 10];
foreach (var row in input.Split('\n'))
{
    j = 0;
    foreach (var col in row.Trim().Split(' '))
    {
        result[i, j] = int.Parse(col.Trim());
            j++;
        }
        i++;
    }


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Home Sports Usain Bolt's farewell race in Jamaica this Saturday: Racers Grand Prix
Usain Bolt's farewell race in Jamaica this Saturday: Racers Grand Prix
Usain Bolt will be honoured at could be an emotional 2nd annual Racers Grand Prix at the National Stadium in Kingston this Saturday, June 10.
Jamaica welcomes the world to the highly anticipated track event, which will be hosted under the theme 'Salute to the Legend', to honour the career of the athletics icon, the greatest-ever sprinter, who will be retiring at the end of this season.
Usain Bolt's farewell at Racers Grand Prix
It may be the last opportunity for many Jamaicans to see Usain Bolt run in a local competitive race and also to say thanks and farewell to the legend before he hangs up his spikes after the World Championships in London August 4–13.
Some of the world's finest athletes have arrived in Jamaica this week to compete and pay homage to the legend, including world record holders South African Wayde van Niekerk (400m) and Kenyan David Rudisha (800m). Bolt's very good friend Somalian-born Mo Farah of Britain will also be in action.
With the help of the Government of Jamaica and sponsors, the Racers Track Club will pay tribute to its most outstanding athlete to date, Usain Bolt, a 9-time Olympic champion, 11-time world champion and the world record-holder for the 100 and 200 metres.
Racers Grand Prix


Law and Social & Public Policy focuses on social problems such as poverty, homelessness and ill-health. The programme applies ideas from political science, sociology and economics to explore how governments shape their responses, and to understand the impacts of public policy on society.
This intellectually stimulating degree addresses the broader aspects of business organisation and management culture, while also providing an insight into corporate and consumer legislation, corporate regulation and other legal aspects of business enterprise. Also, this course will equip you with a real understanding of how legal considerations have an impact on virtually every aspect of management activity.
On this course you will usually be taught by a range of staff with relevant expertise and knowledge appropriate to the content of the unit. This will include senior academic staff, qualified professional practitioners and research students. You will also benefit from regular guest lectures from industry.
This professional law degree course is specifically designed to give you the skills employers are looking for. Whether you are considering a career in law, business or a related field, our degrees equip you with the all-important business skills you need to succeed. Research informs teaching at every level and you will investigate and develop core legal knowledge in subjects spanning commercial law, criminal justice, environmental law, human rights and family law.
Thanks to this course, you will learn the understanding of the origins of and responses to criminal behaviour. This criminology degree will allow you to gain a wider understanding of this hotly debated political issue. It covers specialist areas that include youth violence and gangs, organised crime, and terrorism and security, as well as developing their understanding of criminological theories including the assumptions that underlie intervention and control. You'll look in-depth at the Criminal Justice System, including the police, judiciary and prisons, examining the concept of justice and sentencing.
The Bachelor in Criminology is a diverse, stimulating and challenging subject. It covers many disciplines such as psychology, sociology, social policy, law, biology in order to investigate high social problems of our time: What are the causes of crime? What should we do about it? How best can we support victims of crime?
The Youth and Community Work Bachelor will allow you to work with children and young people in a range of fields: helping them to enhance their personal, social and educational development. Throughout the Youth and Community Work programme you will develop your knowledge and skills to develop a critical, analytical and reflective thinking.


text:
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text:
Trichogenia maxillaris is een haft uit de familie Heptageniidae. De wetenschappelijke naam van de soort is voor het eerst geldig gepubliceerd in 1988 door Braasch & Soldán.
De soort komt voor in het Oriëntaals gebied.

Heptageniidae
Dier uit het Oriëntaals gebied


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Human Faces Can Express at Least 21 Distinct Emotions
Happily surprised
Image courtesy of The Ohio State University.
By Alice Park
Leading scientific thinkers of their time, such as Aristotle, Rene Descartes, Guillaume Duchenne, and Charles Darwin, have long promoted the idea that there are a handful of basic emotions that people express. In recent decades, that group has crystalized into six core emotions: happiness, surprise, sadness, anger, fear, and disgust.
But there are clearly many shades of gray between those emotions. For example, there's the happy-because-I'm-eating-ice cream and the happy-because-I-just-learned-I-got-a-surprise-marriage-proposal looks, each of which is slightly different.
That's what intrigued Aleix Martinez, associate professor of electrical and computer engineering at Ohio State University. "Six seemed a small number given the rainbow of possibilities of feeling and expressing emotions," he says.
MORE: Emotions May Not Be So Universal After All
Martinez wanted to know whether compound emotions, such as happy surprise, were expressed using the same muscle movements of both happiness and surprise, or whether the expression involved a unique set of muscles that represented some amalgam of the two.
Get our Health Newsletter. Sign up to receive the latest health and science news, plus answers to wellness questions and expert tips.
What he and his colleagues found was that the human face makes 21 different emotional expressions – and each is different from the other. While some represented combinations of emotions, each differed in terms of which muscles were involved.
And surprisingly, these facial expression patterns were remarkably consistent across all 230 volunteers. For example, each showed happy surprise in the same way that was distinct from both happiness and from surprise, and different still from angry surprise.
MORE: To Really Read Emotions, Look at Body Language, Not Facial Expressions
Martinez broke down the facial expressions of 230 volunteers by applying his engineering strategies. He and his colleagues gave each of the students, staff, or faculty members who enrolled in the study different scenarios and asked them to show how they would react in each one. They were told, for example, that they had just learned they had been accepted to a graduate program, that someone had told them a disgusting, but still funny joke, or that they had just smelled something bad. The volunteers were allowed to practice their facial expressions in front of a mirror before Martinez took pictures of their reactions.
He then computer-analyzed each of the 5,000 images, breaking them down by which facial muscles the participants used. These were first defined in 1978 by psychologist Paul Ekman, who codified facial expressions in the Facial Action Coding System (FACS) by action units, or muscles or groups of muscles that went into making facial expressions – such as lip parts (for showing disgust), showing teeth (for expressing happiness), mouth stretch (for fear), or eyelid tightening (for anger).
MORE: How to Lift Your Mood? Try Smiling
Not only are people able to make compound expressions consistently, but we are also able to read these emotions pretty accurately as well. When we see someone who looks angrily surprised, for example, we know that they aren't happy about whatever unexpected event they just experienced. "We don't know for sure how much is learned or innate in expressing these emotions," says Martinez. "But what we do know is that a big component has to be innate because otherwise different people would use different muscles to show the same emotion."
The results dovetail with recent findings that some cultures may already recognize these more refined, and numerous expressions of emotions. In a study among the remote Himba tribes in Namibia, for example, researchers at Northeastern University reported that when they provided tribe members with images of facial expressions of emotions, rather than creating six neat piles of emotions, the members created many more. For them, happy could be interpreted as anything from happy to laughing to wonder.
That should help to improve how studies of human emotions are conducted in different cultures, says Martinez; his results suggest that the underlying ability to express emotions may be similar around the world, but cultural biases may simply define emotions in different ways – much the same way that babies are born with the capacity to speak and make sounds for any language, but are trained to speak their native tongue by what they hear around them.
Even more exciting for mental health experts is the possibility that this work can teach them about when the processing and expression of human emotions goes awry such as in depression. "It's important to understand which are categories of emotions that we have," says Martinez, so that experts can recognize the pathological ones. Expressing emotions is generally a transient exercise, lasting milliseconds or up to a minute. Those that linger longer, for hours or even days, tend to be considered moods, and once these emotions persist for days, weeks or months, they can become the subject of mental illness. So distinguishing how people express themselves, and defining the categories of well-known and well-recognized emotions, could lead to better understanding of which emotions can become detrimental and even harmful.
What to Know About Other Impeached Presidents


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I currently am using a service provider that has registered 3 of my domain names and re-registers them every year. Also I am using the same provider for server space for the websites. I would like to get step by step instructions to migrate these pages to another service provider and gain control over the domain names they are registering for me as I think I can do this for significantly less $. Please let me know if there is any other info you may need.
5) Call your new site provider and ask them their process for switching the registration to their control (I think it requires you to request the site key ID for the transfer of the registration from the old provider, and then giving that to your new one and letting them input it and take control for it).


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St. Paddy  (1957–1984) was a British Thoroughbred racehorse and sire. In 1960, he won both the Epsom Derby and the St Leger. His performances in both 1960 and 1961 were instrumental in making his sire Aureole the Leading sire in Great Britain & Ireland for each year.

Background
Owned and bred by Sir Victor Sassoon, he was out of the mare Edie Kelly, and sired by Aureole, a winner of the Coronation Cup and King George VI and Queen Elizabeth Stakes. Edie Kelly, who ran 14 times winning one small apprentice handicap, later produced Parmeila, a filly who won the Ribblesdale Stakes and the Park Hill Stakes in 1970. St. Paddy's grandsire, Hyperion, and his damsire, Bois Roussel, both won The Derby. St. Paddy was bred and raised at his owner's Thornton Stud, Thornton-le-Street, North Yorkshire.

Racing career

1959:two-year-old season
St. Paddy finished unplaced on his debut in the Acomb Stakes at York in August. In September he won the Royal Lodge Stakes at Ascot.

1960:three-year-old season
On 27 April 1960, St. Paddy finished unplaced behind Martial in the 2000 Guineas on his seasonal debut. He then won the Dante Stakes at York in May.

In the Derby on 1 June at Epsom he was ridden by Lester Piggott and started at odds of 7/1. The race was run in warm, sunny weather and attracted the customary huge crowd,which included the Queen. He was prominent from the start, took the lead three furlongs from the finish, and was never in danger, winning easily by three lengths from Alcaeus with Kythnos third. The race was marred by a fatal injury sustained by the favourite Angers.

He was narrowly beaten in the Gordon Stakes by Kipling, to whom he was conceding five pounds. He then won the Great Voltigeur Stakes at York in August. In the St Leger at Doncaster in September he stated odds-on favourite against eight rivals and won easily by three lengths from Die Hard.

1961: four-year-old season
St. Paddy won the Coombe Stakes at Sandown, the Hardwicke Stakes at Royal Ascot and the Eclipse Stakes at Sandown in July. In the Eclipse he led from the start and won impressively from Proud Chieftain, leading the Glasgow Herald to describe him as "the complete racehorse". He started favourite for the King George VI and Queen Elizabeth Stakes at Ascot one week later, but was beaten three lengths into second place by the Prix du Jockey Club winner Right Royal.

In Autumn, St. Paddy won the Jockey Club Stakes, but was beaten in the Champion Stakes. His defeat meant that he failed to break the record for prize money by a British-trained horse, his final total of $272,141 placing him second behind Ballymoss. He was then retired to stud.

Stud Record
St. Paddy was retired after the 1961 racing season to stand at stud at Beech House Stud in Newmarket. As a sire, he met with reasonable success, notably siring Connaught, winner of the Eclipse Stakes and Jupiter Island, a multiple graded stakes race winner whose biggest success came in the 1986 Japan Cup. St. Paddy was pensioned in 1981 and died on 16 May 1984 at the age of 27. He is buried at Beech House Stud.

Assessment and Honours
Following the London & North Eastern Railway tradition of naming locomotives after winning racehorses, British Railways "Deltic" Diesel locomotive no. D9001 (later 55001) was named after this horse on 7 July 1961, and remained in service until 5 January 1980.

In their book A Century of Champions, John Randall and Tony Morris rated St. Paddy an "average" Derby winner and the one hundred and twenty-first best British racehorse of the 20th Century .

Pedigree

References

External links
Footage of St. Paddy winning the 1960 Epsom Derby

1957 racehorse births
1984 racehorse deaths
Racehorses bred in the United Kingdom
Racehorses trained in the United Kingdom
Thoroughbred family 14-c
Epsom Derby winners
St Leger winners


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The Legend of Zelda: Breath of the Wild DLC Pack 1 Coming June 30
Nintendo detailed the first DLC pack coming for The Legend of Zelda: Breath of the Wild today. Slated for release on June 30, this "Master Trials" pack will include some rather interesting added content. Keep in mind that this is part of the $19.99 (USD) Expansion Pass for the game.
Trial of the Sword: challenges the player to go through waves of enemies in a total of about 45 rooms. By completing the challenge, Link will prove that he has grown worthy, and the
Super Mario Odyssey E3 2017 Trailer and Details
Mario is back on a new 3D adventure the likes of which you haven't seen since Super Mario 64 and Super Mario Sunshine. Super Mario Odyssey finds Mario visiting plenty of diverse locations as he attempts to rescue Princess Peach from marrying Bowser.
Huge 3D kingdoms filled with secrets and surprises, including costumes for Mario and lots of ways to interact with the diverse environments - such as crusing around them in vehicles that incorporate the HD Rumble feature
Metroid Prime 4 Announced for Switch
It wasn't the most prolific announcement but it's one that made tons of fans happy today. Metroid Prime 4 is being developed for the Nintendo Switch.
"Metroid Prime 4 (working title) returns to the first person adventure roots of the original Metroid Prime game and introduces a new storyline that ties together the events of the Metroid Prime universe and takes the storyline in new directions" according to series producer Kensuke Tanabe. Tanabe is leading the project together with
Kirby for Nintendo Switch Announced
Nintendo announced that Kirby is making a big return, this time on the Nintendo Switch. You and three friends can team up when Kirby for Nintendo Switch (working title) releases in 2018.
Recruit up to three enemies as helpers by hitting them with hearts
Returning copy abilities include: Sword, Fire, Water, Bomb, Broom, and more
Imbue copy abilities with different elements such as: Wind, water, fire, and electricity, by borrowing or giving them to helpers to
SEGA and Atlus Showcase Six Titles at E3 2017 - Yakuza, Etrian Odyssey, and More
SEGA and Atlas passed along information and trailer links to six titles they're showing off this week at E3 2017. We have some tasty new media for Yakuza Kiwami, Yakuza 6: The Song of Life, Etrian Odyssey V: Beyond the Myth, Radiant Historia: Perfect Chronology, Shin Megami Tensei: Strange Journey Redux, and Utawarerumono: Mask of Truth. So, let's just right into it.
Yakuza Kiwami In 2005, a legend was born on the PlayStation 2 - the legend of Kazuma Kiryu, the Dragon of Dojima, in hi
Final Fantasy XV Expanded Universe Showcased in New Videos
Square Enix used E3 to showcase some of the content coming to both the base game of Final Fantasy XV and some new projects they're working on to expand the "Final Fantasy XV Universe" as they're calling it.
First up, the additions coming to the main game.
Support of next level hardware
GTA Online: Gunrunning Now Available
Rockstar just released the new Gunrunning update for GTA Online. We're looking at six new weaponized vehicles, new clothing, weapon upgrades, underground bunkers, mobile ops centers, and more.
Dominate the illegal arms trade in Southern San Andreas with Gunrunning, the latest update to Grand Theft Auto: Online. Your career as a prolific arms dealer begins with the acquisition of a Bunker. These underground fortresses serve as your base of operations for all Gunrunning activities,
Call of Duty: WWII Multiplayer Reveal Trailer
There was a multiplayer reveal trailer shown tonight for Call of Duty: WWII. Chance are, if you are interested in this game you already say the trailer since it literally already has 1.1 million views in just a couple of hours. What was that about the franchise supposedly dying?
Destiny 2 - New E3 Trailer, Beta Dates Announced, PC Release Date Announced
First thing is first, Bungie has confirmed the dates for the Destiny 2 beta on the PlayStation 4, Xbox One, and PC.
Destiny 2 BETA
July 18: PlayStation 4 Pre-Order BETA
July 19: Xbox One Pre-Order BETA
July 21: Open BETA for PlayStation 4 and Xbox One
July 23: Console BETA End-date
Late August: PC BETA
They have also confirmed the release dates for all platforms.
Destiny 2 LAUNCH
September 6: PS4 and Xbox One
Horizon Zero Dawn: The Frozen Wilds DLC Announced; Releasing in 2017
If you enjoyed Horizon Zero Dawn as much as I did, you're probably pretty excited to learn that there is some new DLC coming later this year for the game. Horizon Zero Dawn: The Frozen Wilds offers up some new locations, more fights, more story, and is set to be released later this year.
Shadow of the Colossus Remake Coming to PlayStation 4
Sony is once again utilizing the talented individuals over at Bluepoint Games to remake one of their most beloved titles. Yes, Shadow of the Colossus is being remade for the PlayStation 4.
Like many other titles at E3, this one will be released at some point in 2018.
Spider-Man PS4 Gameplay from E3
Spider-Man for the PlayStation 4 finished out the Sony conference tonight at E3. It features a decent chunk of gameplay, which should give you a pretty good idea of what to expect from the full game.
The game is currently set for a vague "2018" release. Also, I hope you don't mind some QTEs in your Spider-Man games!
Marvel vs. Capcom: Infinite - Story Demo Out Now, More Playable Characters Confirmed
Capcom just released the demo for Marvel vs. Capcom: Infinite on both the PlayStation 4 and Xbox One. In addition to this playable demo, the studio also confirmed a handful of additional characters that you will get to play as.
With the release of the new story demo, fans are now able to delve deeper and immerse themselves in MARVEL VS. CAPCOM: INFINITE like never before. This thrilling cinematic experience puts players at the center of a fierce battle as heroes from both universes
Monster Hunter: World Announced for PS4, XB1, and PC
During Sony's conference, Capcom announced Monster Hunter: World for the PlayStation 4. Yes, it's also coming to the Xbox One and PC, relax. You will have some time to wait though as the game isn't scheduled to be released until early 2018 for PS4 and XB1 and even later than that for the PC (ugh).
Uncharted: The Lost Legacy E3 Trailer
Arriving August 22, 2017, Uncharted: The Lost Legacy is a standalone expansion to Uncharted 4. A new story focused trailer came out tonight that shows off Chloe and Nadine getting into a bit of trouble. Keep in mind that this will be free to those that purchased the Season Pass for Uncharted 4 early in the game's release window.


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Barbarism: 22 year -old man refuses to serve deity, escapes from community
December 31, 2012 July 4, 2018 nigerianewspoint
Mr Felis
What appeared a baberic and unbelievable inhuman act was recorded last weekend, December 29, 2012 in the South South geopolitical zone of Nigeria, as a twenty two year old man was forced out of his fatherland for not serving a deity.
The victim identified as Felis Innocent, who hails from Ivue in Uromi Edo State, Nigeria, and a Primary 4 school dropout has reportedly left his original abode uncelebrated, when he declined to serve the village deity, served by his late father, Mr. Felis.
Information gathered by our roving reporter said the deceased Chief Priest, Mr Felis who died in December 20, 2012 after serving the shrine for over 13 years left behind his wife and only child, Innocent.
Further investigation revealed that according to the traditions of the Ivue people, once a Chief Priest dies, his male offspring automatically takes over as a successor.
One week after his burial according to the traditional rites and rituals, the villagers through the traditional ruler and his elders in council demanded that Innocent be installed as the next Chief Priest.
When contacted, the only son, Felis Innocent refused ascending the position by telling them it was against his Christian faith.
Angered by his refusal to continue with his father's footprint of serving the shrine, they allegedly threatened to use him as an appeasing sacrifice to their gods or they would banish him.
Insisting on keeping to his Christian faith, and due to fear of being killed by the angry village chiefs, the young man reportedly left his village to an unknown destination as at press time.
Reacting to her son's whereabouts, and circumstances surrounding her son's disappearance, Mrs Felis who couldn't hold back her tears said, "Since they threatened to kill my only son because of his refusal to be an idol worshipper, I have not set my eyes on him. Who knows if he is still alive or dead? He is my only child, who I still live for".
She continued by telling newsmen that even when her husband was alive, she and her disappeared son never took part in idol worshipping because it was against their faith.
Meanwhile, a cross section of people interviewed on the issue blamed the government on the negligence of fundamental human rights of the citizens.
Man,18, escapes death for refusing to serve deity …Family banished, house burnt


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Club Atlético Basáñez  is a football and boxing club from Montevideo in Uruguay founded in 1920. They currently play in the Second Amateur Division, the third and last tier of the Uruguayan championship.

The club is named after Tomas Basáñez, who was a major landowner in the locality of the club.
The club colors are inspired by those of the anarchist flag.
One of the most popular teams from the lower divisions along with Cerrito and Racing.

First Division
After being champions of the Uruguay Second Division in 1993, they were promoted to play in first division where they spent two seasons 1994 and 1995. They had never played in First Division before.
Prior to this, the club gained promotion to first division in 1992, but lost its place because of points reduction, due to disturbances between the fans and horse-mounted police.

Titles
Uruguayan Primera División: 0
Amateur Era (0):
Professional Era (0):

Segunda División Uruguay: 1
1993

Tercera División Uruguay: 1
1989

Special kits

External links

Basáñez profile at futbolvida.com.uy

Football clubs in Uruguay
Association football clubs established in 1920
Sport in Montevideo
1920 establishments in Uruguay