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Cult Fashion Store Opens In East London
Story from Fashion
Cult Fashion Store Browns Just Opened A New Space In East London
Alice Casely-Hayford
Over the past decade, Shoreditch has been transformed into a creative hub and more recently, a must-visit shopping destination with the boom of independent fashion stores around Shoreditch High Street, Redchurch Street and Calvert Avenue. This week, it became even more desirable for style aficionados and shopping enthusiasts as cult luxury fashion mecca Browns opened the doors to Browns East, its first bricks and mortar store to open for 20 years. Located on the corner of Club Row, the shiny new two-storey space, housed in a former print factory, complements the flagship store on South Molton Street, first established in the heart of London's Mayfair in 1970.
Offering a completely fresh concept for its customers, Browns East is an augmented retail environment, which will be constantly changing and flexible. What does that actually mean? I hear you ask. Well, the entrance, an area in its own right, will regularly be handed over to independent creatives to build an immersive space, allowing Browns to further support and champion fresh talent in an authentic way. The inside space will also include the first incarnation of the Nomad project, Browns' take on semi-permanent, roaming retail concepts as a 21st-century response to the "pop-up store" model.
Holli Rogers, CEO of Browns explains: "Browns has a strong heritage in innovation; from the time we first opened our doors in 1970, Browns has showcased pioneering designers. We've blended our love of fashion with incredible in store experiences and we've always ensured that we change and evolve with our customers. The store will have new elements that are tied to emotion and identity – it's truly a sensory experience. The moment you walk in you will feel something – through the people, the product, the design, the areas within the space, and the events that will take place. Coupled with this, we are innovating with technology which will make the shopping experience in Browns more personal and convenient for our customers. The vibrancy of East London is great – it's a neighbourhood of immense creativity and boldness. Opening a store here is an exciting move with a new view on what retail looks like."
The clothes and accessories will be presented in a gender-neutral retail space, where barriers between menswear and womenswear don't exist. Offering a regularly rotating edit of the best in luxury fashion, across ready-to-wear, footwear, bags and fine jewellery, Browns East launches with 45 brands, including some of our favourites such as Rejina Pyo, Jacquemus, Gucci and Sies Marjan.
And when you need a moment's respite after perusing the rails there's a café with a marble-clad bar, stocked for launch by Fatties Bakery, where you can pick up a shiny cornflake bocadito or salted caramel brownie and turmeric latte.
Browns East also has impressive eco credentials; floorboards originally laid at the BBC have been carefully preserved and used in the ground floor space, while striking fixtures and furniture have been crafted from 100% recycled plastics.
The staircase-meets-gallery is home to a curated selection of multimedia artwork with launch artists including Polly Morgan, Juno Calypso and Lauren Baker, and all pieces are available for purchase at a range of price points.
As the closing date of iconic concept store Colette looms (December 20th if you want to book an express shopping trip to Paris), the opening of another branch of Browns is exactly the kind of exciting fashion news we've been waiting for.
Browns Fashion Opens New Store Shoreditch East London
Fashion • Shopping
written by Alice Casely-Hayford
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Home >Blog >Facebook Marketplace Takes on Giants eBay, Craigslist
Facebook Marketplace Takes on Giants eBay, Craigslist
David Bosley 6 December 2016
Whether it's instant messaging, gaming, photo sharing, advertising, or catching up on the latest news, it seems like Facebook does it all. But the social media giant just added one more service to its arsenal, and according to Forbes magazine, it's going to make a big splash this holiday shopping season.
With the introduction of Facebook Marketplace, the site is taking on sites like eBay and Craigslist in peer-to-peer commerce platforms. Experts argue that the end goal with Marketplace is obvious – keep users on Facebook longer to eliminate the need to visit other sites.
Facebook's goal has some significant implications for digital marketing professionals, but perhaps most importantly is how crucial it is for brands to have a strong social media presence to ensure conversions from page to Marketplace.
Read more at Forbes.com
Facebook Marketplace Takes on Giants eBay, Craigslist was last modified: April 25th, 2019 by David Bosley
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redpajama_set_name: RedPajamaCommonCrawl


You know how we often regret of the things we've done in the past and think that we'll do it differently if we get a second chance?
I've recently watched this movie called "The Vow". It's about this girls who got into an accident and woke up not remembering the more recent part of her life including her marriage and her husband. At the end, she still couldn't remember the lost part of her life but somehow she made the exact same decision about her life all over again just like she did before the accident.
It makes me realise, a second chance without the wealth of experience is useless. So it's no use going back to the past wanting to redo what we've done because even if we can, unless we remember that we've done it and learn from the mistakes, the chance is we will still do the same thing all over again.
So moral of the story, never dwell in your past mistakes because they're the most important lessons. Move on and utilise this knowledge to do better.
This entry was posted in Movies, Opinions, Personal. Bookmark the permalink.


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DTC Index
Tamara Mellon Brand raises USD$50m (£39.28m); USD$13m (£10.2m) funding round for AUrate
by	Mathew Broughton on	26th Jun 2019	in	News
DTC's Daily Digest brings you the latest news on the world's fastest growing direct-to-consumer brands. In today's edition: Tamara Mellon Brand raises USD$50m (£39.28m), with sales increasing by 136% year-on-year; USD$13m (£10.2m) funding round for AUrate; and Grab invests USD$8m (£6.3m) in London startup Splyt.
Tamara Mellon Brand raises USD$50m (£39.28m), with sales increasing by 136% year-on-year
Tamara Mellon Brand, a luxury DTC women's footwear company, has announced it has raised USD$50m (£39.28m) in a Series C funding round led by investment firm Centricus, bringing total funding in the company up to approximately USD$87m (£68.4m). The funding is reportedly due to be used to support manufacturing, brand growth, and expanding the team to a total of 60 employees by the end of the year. The company, led by the eponymous Tamara Mellon, who co-founded the Jimmy Choo brand, also announced that sales had increased by 136% year-on-year.
"Tamara and I are thrilled to have Centricus join our impressive group of investors," said Tamara Mellon Brand co-founder Jill Layfield. "They are a smart and thoughtful team with an incredibly important global perspective. We are also very grateful to our customers and employees who have been with us for an amazing two-and-a-half years, and can't wait for what comes next."
USD$13m (£10.2m) funding round for AUrate
DTC jewellery brand AUrate has announced a fundraising round of USD$13m (£10.2m), aimed at expanding the company outside of its US heartland into Europe and the Middle East. Investors involved in the round included the Arab Angel Fund, Bluecrest Capital, and Point King Capital. While the company does not publicly disclose its revenue, it has revealed that it has grown by nearly 400% since a USD$2.6m (£2.04m) funding round which took place in 2017.
Speaking on the plans to expand AUrate, co-founder Bouchra Ezzahraoui said, "if you want to become a real luxury brand, you have to have that international appeal. When you talk to investors, they ask what will the brand look like in 10 years. Anyone can open a store online and start selling here, but luxury brands are all international. The fact that all of our stuff is made in the U.S. is a stamp of trust, but we want to be a global brand."
Grab invests USD$8m (£6.3m) in London startup Splyt
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Your Instagram Is An Accurate Test for Depression
Your Instagram Account Could Be a Scary Accurate Indicator of Depression
Science says certain clues in your Instagram photos could give away your mental health status.
By Charlotte Hilton Andersen Published: Aug 17, 2016
Instagram is a great way to share your vacations, passions, and sense of humor, but your account may be broadcasting something else as well: your mental health status. Turns out, the way depressed people see the world is reflected in the type of photos they choose to take and share on social media. These differences can be picked up by a new artificial intelligence program, helping the computer recognize which users are showing signs of depression, according to new research from Harvard University's Andrew Reece and the University of Vermont's Christopher Danforth. The strangest part? It turns out the computers are better at recognizing this human emotion than actual humans are.
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The researchers gathered 44,000 Instagram photos from 166 different users — some of whom had been diagnosed with depression while the rest were mentally healthy — and asked normal people (i.e. not psychologists) to analyze the pictures; they also ran the snaps through their computer program. The people ranked the pictures according to emotion, but the machine used a complex algorithm to examine the photo's color, composition, saturation, number of people, and even the type of Insta filter used. The machine was able to pick out the depressed people with a much higher degree of accuracy than the real people.
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What gave the depressed user away? As you might guess, the pictures of people with depression were darker hued, had more blues and grays, and were less bright, suggesting a bleaker outlook. But there were some surprises. The more comments a post received, for example, the more likely the poster was depressed — yet the more likes a post got, the less likely the poster was depressed. (The study didn't say what it meant if you get a lot of likes and comments.) The choice of filter was also a big clue, with "Valencia" being the most popular among non-depressed individuals and "Inkwell" being the top pick for depressives.
And then there were the number of people in the photo. Those suffering from depression were surprisingly more likely to post more pictures of faces, but the number of people in each shot was lower, perhaps indicating a smaller social circle.
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THE DEATH OF STALIN: GENIUS AT WORK
Film In Theaters Leonard Maltin Movie Reviews March 9, 2018
I consider writer-director Armando Iannucci to be a comic genius. The creator of Veep cut his teeth in British TV comedy working with Steve Coogan on his Alan Partridge character, then created a series called The Thick of It, which like its American offspring is foul-mouthed and laugh-out-loud funny. He bridged the two series with a first-rate feature film about English and American politics called In the Loop. Now comes an even more ambitious endeavor, The Death of Stalin, based on a French comic-book series depicting the aftermath of the notorious Soviet dictator's demise. It is brilliant and hilarious, a delicious mélange of political satire and farce.
The premise is straightforward: Josef Stalin dies by choking in his office one night. When his body is discovered the next morning, his closest colleagues gather and frantically jockey for power, exposing their own peccadilloes while sabotaging each other's chances to become Russia's supreme leader.
Like Veep, this movie depends on great writing, expert casting, and unimaginable rehearsals to refine the timing, delivery and staging that make the most of a knockout screenplay. (Iannucci wrote the script with longtime colleagues David Schneider, Ian Martin, and Peter Fellows, based on a screenplay by Fabien Nury.) The filmmakers don't assume we know anything about the history they are depicting; you don't have to take an entrance exam to enjoy their movie. What's more, they aren't afraid to be outlandish or silly.
The cast is daunting, to put it mildly. It wouldn't have occurred to me to choose Steve Buscemi to play Nikita Krushchev, but he is superb, an adjective that applies equally to Jeffrey Tambor, Simon Russell Beale, Michael Palin, Jason Isaacs, Rupert Friend, Paddy Considine, Olga Kurylenko, Andrea Riseborough, and the other members of the ensemble. Beale, primarily a stage actor, is overwhelming in his ferocious interpretation of Levrenti Beria, the wiliest and nastiest man in the Soviet Politbureau. Adrian McLoughlin sets the movie on its comedic course with his wholly unexpected take on Stalin.
I have seen this movie twice in six months' time and would readily watch it again after taking a breather. Iannucci conducts the proceedings like the conductor of a great orchestra with an array of virtuosos at his command. The Death of Stalin is a breath of fresh air and a genuinely great screen comedy.
Tagged Adrian McLoughlinAlan PartridgeAndrea RiseboroughArmando IannucciDavid SchneiderFabien NuryIan MartinIn The LoopJason IsaacsJeffrey TamborMichael PalinOlga KurylenkoPaddy ConsidinePeter FellowsRupert FriendSimon Russell BealeSteve BuscemiSteve CooganThe Thick of ItVeep
Leonard Maltin
Leonard Maltin is one of the world's most respected film critics and historians. He is best known for his widely-used reference work Leonard Maltin's Movie Guide and its companion volume Leonard Maltin's Classic Movie Guide, now in its third edition, as well as his thirty-year run on television's Entertainment Tonight. He teaches at the USC School of Cinematic Arts and appears regularly on Reelz Channel and Turner Classic Movies. His books include The 151 Best Movies You've Never Seen, Of Mice and Magic: A History of American Animated Cartoons, The Great Movie Comedians, The Disney Films, The Art of the Cinematographer, Movie Comedy Teams, The Great American Broadcast, and Leonard Maltin's Movie Encyclopedia. He served two terms as President of the Los Angeles Film Critics Association, is a voting member of the National Film Registry, and was appointed by the Librarian of Congress to sit on the Board of Directors of the National Film Preservation Foundation. He hosted and co-produced the popular Walt Disney Treasures DVD series and has appeared on innumerable television programs and documentaries. He has been the recipient of awards from the American Society of Cinematographers, the Telluride Film Festival, Anthology Film Archives, and San Diego's Comic-Con International. Perhaps the pinnacle of his career was his appearance in a now-classic episode of South Park. (Or was it Carmela consulting his Movie Guide on an episode of The Sopranos?) He holds court at leonardmaltin.com. Follow him on Twitter and Facebook; you can also listen to him on his weekly podcast: Maltin on Movies. — [Artwork by Drew Friedman]


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 Tar River, een rivier in North Carolina
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redpajama_set_name: RedPajamaWikipedia


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Traumatic brain injury causes selective, CD74-dependent peripheral lymphocyte activation that exacerbates neurodegeneration
Richard P Tobin1,
Sanjib Mukherjee1,2,
Jessica M Kain1,2,
Susannah K Rogers3,
Stephanie K Henderson4,
Heather L Motal1,
M Karen Newell Rogers1,4,5 &
Lee A Shapiro1,2,4
Acta Neuropathologica Communications volume 2, Article number: 143 (2014) Cite this article
Traumatic brain injury (TBI), a significant cause of death and disability, causes, as in any injury, an acute, innate immune response. A key component in the transition between innate and adaptive immunity is the processing and presentation of antigen by professional antigen presenting cells (APCs). Whether an adaptive immune response to brain injury is beneficial or detrimental is not known. Current efforts to understand the contribution of the immune system after TBI have focused on neuroinflammation and brain-infiltrating immune cells. Here, we characterize and target TBI-induced expansion of peripheral immune cells that may act as potential APCs. Because MHC Class II-associated invariant peptide (CLIP) is important for antigen processing and presentation, we engineered a competitive antagonist (CAP) for CLIP, and tested the hypothesis that peptide competition could reverse or prevent neurodegeneration after TBI.
We show that after fluid percussion injury (FPI), peripheral splenic lymphocytes, including CD4+ and CD8+ T cells, regulatory T cells (Tregs), and γδ T cells, are increased in number within 24 hours after FPI. These increases were reversed by CAP treatment and this antagonism of CLIP also reduced neuroinflammation and neurodegeneration after TBI. Using a mouse deficient for the precursor of CLIP, CD74, we observed decreased peripheral lymphocyte activation, decreased neurodegeneration, and a significantly smaller lesion size following TBI.
Taken together, the data support the hypothesis that neurodegeneration following TBI is dependent upon antigen processing and presentation that requires CD74.
Traumatic brain injury (TBI) is a leading cause of death and disability in the United States and throughout the world [1]-[3]. To date, there are no effective treatments to prevent or reverse damage after TBI. Due to the direct impact on the brain, efforts at diagnosis and treatment of TBI have been primarily focused on the central nervous system (CNS). However, it is now well established that peripheral immune mechanisms are involved in the pathology associated with TBI [4]-[7]. The paradigm that has been widely accepted is that the blood brain barrier (BBB) serves as the primary obstacle to the recruitment of leukocytes that could otherwise cause harm to the brain [7],[8]. However, after TBI, there is BBB breakdown and one result is the passage of peripheral immune cells into the CNS [7].
In general, an immune response will begin with an innate and immediate response to injury that includes activation and migration of neutrophils, dendritic cells, macrophages (including microglial cells in the CNS), as well as other less defined cellular and molecular components [9]-[11]. The innate immune response is typically followed by the antigen-specific, adaptive immune response. The transition from an innate to an adaptive response is dependent upon successful antigen processing and presentation, by professional antigen presenting cells (APCs), to antigen-specific T lymphocytes [12],[13]. How, or if, damage to the brain, often described as an "immune-privileged" site [14], elicits an innate, and the transition to, an adaptive immune response from peripheral immune mediators to the CNS, is not known. Evidence for the involvement of antigen processing and presentation in the CNS is provided by the study of Slavin et al. [15]. This study demonstrated that mice lacking the machinery for processing and presentation of full-length myelin basic protein did not develop experimental autoimmune encephalomyelitis (EAE), a mouse model for Multiple Sclerosis (MS). However, when these same mice were exposed to peptides derived from full-length myelin basic protein, the mice became susceptible to EAE. Interestingly, Ling et al. demonstrated that TBI results in recruitment of antigen-specific CD8+ T cells, only in the presence of the antigen for which they are specific [16]. Furthermore, in a model of optic neuritis, the acute immune response leads to an adaptive, specific, anti-brain response [17]-[19]. Conflicting reports suggest that this effect could be either harmful or helpful to recovery [5],[6]. Regardless of their role in the outcome, these findings provide evidence for an adaptive immune response in the brain after TBI.
Efficient antigen processing and presentation via the major histocompatibility complex II (MHCII) pathway in APCs, including macrophages, microglia, dendritic cells, B cells, and γδ T cells, facilitates the transition between innate and adaptive immunity [13],[20]-[22]. Antigen processing and presentation by these professional APCs involves invariant chain (CD74). CD74 serves as a chaperone for MHCII until CD74 is proteolytically cleaved into Class II invariant peptides (CLIP) that fill the antigen-binding groove of MHCII molecules [12],[23]. Replacement of CLIP by antigenic peptides and T cell recognition of antigenic peptide in MHCII is the first step in T cell activation, and the first event in a T cell-mediated adaptive immune response [13],[20],[24]. This vital step of peptide exchange between innate and adaptive immunity has not been elucidated following TBI. Critical to this process is the cleavage of full-length CD74 into CLIP, the loading of CLIP into the antigenic binding groove of MHCII, followed by the replacement of CLIP with antigenic peptides [23]. Therefore, we engineered and synthesized a peptide that competitively antagonizes the antigen-binding site of MHCII, to determine if antagonism of CLIP/antigenic peptide exchange might prevent or reverse the detrimental effects of TBI [25]. Thus, this study was designed to determine if exchanging CLIP for antigenic peptide is important in secondary neurodegeneration following TBI.
Treatment with competitive antagonist peptide (CAP)
CAP was synthesized as previously described [25]. Briefly, using computational design, we identified a 9mer peptide, with 8 total amino acid flanking regions, that by, peptide binding analysis software (MHCPred and netMHC), was predicted to have a higher binding constant than the invariant peptide CLIP for the peptide-binding groove of known MHCII alleles. CAP was synthesized by Elim Biopharmaceuticals. The mice were injected intraperitoneally (i.p.) (1 mg/kg) with CAP. CAP was initially dissolved at 5 mg/mL in dimethyl sulfoxide (DMSO), after which 5 μl of CAP dissolved in DMSO was further diluted with 195 μl of sterile saline and injected (I.P.). In vehicle injected mice, an injection containing 5 μl DMSO dissolved in 195 μl of sterile saline.
6-week-old male C57BL/6 J (N = 16) were purchased from Jackson Labs. Invariant chain deficient (CD74Def) mice (C57BL/6 background; N = 16) were purchased from Jackson Labs and bred at the Scott and White Healthcare animal facility to maintain homozygous CD74 deficiency. Because littermates are not available to compare with the CD74Def mice (as they are bred to homozygosity), we have used age and sex matched C57BL/6 mice (the background strain for the mice). All mouse care, housing, and experimental procedures were approved by the Baylor Scott and White Healthcare Institutional Animal Care and Use Committee, reference number 2013-021. Wild type mice were divided into four groups for flow cytometry and multiplexing and sacrificed at 24 hrs after TBI. These four groups were: sham + vehicle (N = 6); sham + CAP (N = 5); FPI + vehicle (N = 7 WT); FPI + CAP (N = 6). For the CD74Def mice, two groups were used. These two groups were: Sham (N = 7 CD74Def) and FPI (N = 6 CD74Def). In addition, we used separate mice for histological studies and these mice were sacrificed 3 days after TBI (N = 6 WT sham + vehicle, 5 FPI + CAP, and 5 CD74Def). The three day post-TBI time point was selected because our previous study indicated that this is when the peak of TBI-induced neurodegeneration was observed [26].
Fluid percussion injury (FPI)
FPI was induced as previously described [26]. Briefly, mice were anesthetized with isoflurane, prepped, cleaned, and shaved, then put into a stereotactic instrument (Stoelting, Inc.). A 2 mm craniotomy was made over the left parietal cortex making sure to keep the dura intact. The female end of a luer lock syringe was cemented over the craniotomy and attached to the fluid percussion apparatus. A 12 16 ms FPI was delivered at a pressure of ~1.4-1.6 atm. Sham mice received identical treatment except the pressure pulse was never delivered. After injury or sham, suture was used to close the scalp over the wound and mice were returned to their home cage resting on a heating pad. Mice were monitored to ensure that they resumed walking, feeding, drinking and grooming behavior.
Mice were sacrificed and spleens were removed. The tissues were dissociated, single cell suspensions prepared, and red blood cells were lysed using GEY'S buffer [27]. The dissociated splenocytes were treated with FC Block (BD Bioscience) and then stained with the following antibodies; CD3, CD4, CD8, CD25, MHCII (clone M5/114.15.1), B220, γδ-TCR (BD Bioscience), and CLIP (15G4, Santa Cruz Biotechnology). For regulatory T cell (Treg) staining we used the eBioscience mouse regulatory T cell staining Kit according to the manufacturer's directions. Live cells were assessed using the Life Technologies LIVE/DEAD® Fixable Aqua Dead Cell Stain Kit according to the manufacturer's directions. Splenocytes were analyzed using a BD FACS Canto II flow cytometer and data were analyzed using FlowJo software (TreeStar Inc.). See Additional file 1: Figure S1 for gating strategy.
FluoroJade C method
Fluorojade C staining took place as previously described [26]. Briefly, every 6th 50 um section was analyzed within the margins of the lesion. This corresponded to 5-7 sections per mouse. Sections were mounted onto gelatin-coated slides and FluoroJade C histology was performed as previously described [26]. The slides were allowed to air dry, after which cover slips were applied using Vectashield with DAPI (Vector Labs). Prior to analysis, slides were coded to ensure that the reviewer was blind to the condition of the mice. Sections were then visualized using an Olympus IX81 (Olympus Inc., Tokyo, Japan) inverted microscope equipped to visualize FITC. We used the following Olympus UplanS objectives to capture the images: 10× NA0.40 ∞/0.17 FN26.5; 20× nA0.75 ∞/0.17 FN26.5; 40× NA0.75 ∞/0.17 FN26.5. Images were captured using the FV1000 and the Fluoview software (V. 1.7.1.0). Images were saved as 24 Bit Tiff files and brightness and contrast were adjusted using Adobe Photoshop (V. 12.0). For quantitative analysis of Fluorojade C labeling, the peri-lesion area was traced and 400 μm2 grids were randomly placed throughout the traced area. All cells which fell within the grid, or contacted the left and upper borders of the grid were included in the counts, whereas those that touched the right or lower border were excluded. The peri-lesion area consisted of +/- 1 mm anterior/posterior (AP) from the injury focus (AP +1.5 mm; medio-lateral: 1.2 mm).
Measurement of lesion size
Superficial lesion size was measured using a digital caliper (Mitutoyo 150 mm, .01 mm accuracy) as previously described [28],[29]. In order to confirm superficial lesion measurements made with the digital caliper, we assessed the margins of the lesion in the anterior posterior plane, and calculated the expanse of the lesion using a mouse atlas [30]. Initially, every 6th 50 μm Nissl stained section was used for determination. However, in some cases, it was necessary to examine serial sections in which Fluorojade C (or other stains not reported in this manuscript) was stained, in order to confirm that we had accurately accounted for the most anterior and posterior margins of the lesion. It should be noted that considering the margin of error of the digital caliper (100 μm) and the distance between brain slices in the mouse atlas (100 140 μm), all measurements were within the margin of error of these 2 sources.
Cytokine analysis
Fresh, non-perfused tissue was isolated from the ipsilateral cortex including the lesion area and flash frozen in liquid nitrogen. Frozen tissue was homogenized following the manufacturer's instructions (Milliplex MAP kit, Millipore). Protein was estimated with a Bradford assay and similar concentrations were made such that 25 μl of homogenate was added to 25 μl of assay buffer. Then, 25 μl of magnetic beads coated with specific antibodies (MCYTOMAG-70 K-PMX, Milliplex MAP Kit, Millipore) was added to this solution and the reaction was incubated at 4 C for 24 hours. Next, the beads were washed and incubated with 25 μl of biotinylated detection antibody at room temperature (RT) for 2 hours. Finally, 25 μl of Streptavidin–Phycoerythrin conjugate compound was added and incubated for 30 min at RT. The beads were then washed and incubated with 150 μl of sheath fluid for 5 min at RT. The concentration of the analytes is then determined by Bio-Plex Manager software version 5.0. The assays were run in triplicate to confirm the results. Analytes were normalized to total protein concentration. In cases where the cytokines in the brain tissue of three or more mice for a group were below the detection threshold, the data were not presented in the graphs.
Statistical analysis was performed using GraphPad Prism 6 (GraphPad Software Inc.). For comparisons between splenocytes from C57BL/6 J and CD74Deff, a t-test was used with a significance cut-off of P <0.05. For all other analysis, repeated measures ANOVA was used with post-hoc planned comparisons using Dunnett's correction factor.
Expansion of peripheral lymphocytes following Fluid Percussion Injury (FPI)
To characterize the changes in peripheral immune cells following TBI, splenic lymphocytes were harvested at 24 hours after FPI and analyzed using flow cytometry. The results showed that the overall splenic cellularity was significantly increased (P < 0.017) in FPI-treated mice compared to sham-treated controls and this effect was prevented by administration of competitive antagonist peptide (CAP) (Figure 1A). While there was a trend toward increased B220+ B cells (Figure 1B), there was a significant (P < 0.03) reduction in B cell numbers following CAP treatment after FPI (Figure 1B). In addition, there was a significant (P < 0.001) FPI-induced increase in the total number of CD3+ T cells that was significantly (P < 0.001) reduced by CAP treatment after FPI (Figure 1C). Specifically, CD4+ T cells were also significantly increased in number (P < 0.014) and their numbers were significantly reduced (P < 0.018) following CAP treatment (Figure 1D). There were also significant increases in Tregs (P < 0.018), as defined by CD3+, CD4+, high CD25 and expression of the transcription factor FoxP3 (Figure 1E), γδ T cells (P < 0.0.14); (Figure 1F) and CD8+ T cells (P < 0.02); (Figure 1G), all of which were significantly reduced by treatment with CAP (Figures 1E-G). In contrast, there was no significant increase in the numbers of conventional activated CD4+ T cells, nor was there a significant reduction of these cells following CAP treatment (Figure 1H).
Selective expansion of splenic immune cell subsets in response to FPI. (A) The number of viable cells in the spleens (cellularity) of C57BL/6 mice 24 hours following sham surgery or FPI. TBI significantly increased the number of viable cells. (B) The number of B cells (B220+) showing a trend toward B cell expansion after FPI that does not reach significance. However, treatment with CAP significantly reduced B cells after FPI. (C) Analysis of T cells (CD3+) revealed a significant expansion after FPI that is significantly reduced following CAP treatment. (D) A subset of CD3+ T cells, CD4+ T cells (CD3+CD4+), are also significantly elevated after FPI. This elevation is significantly reduced following CAP treatment. (E) Identification of Tregs (CD3+CD4+CD25HiFoxP3+) also revealed a significant increase in this cellular population after TBI. Similar to other T cell populations, this population is also significantly decreased after CAP treatment. (F) A similar pattern of FPI-induced increase is observed for γδ T cells (CD3+γδ-TCR+) and this increase is significantly reduced by our CAP treatment. (G) Analysis of CD8+ cytotoxic T lymphocytes (CD3+CD8+) also revealed a significant increase after FPI that is significantly reduced with our CAP treatment. (H) Alternatively, no significant differences were observed for activated CD4+ T cells (CD3+CD4+CD25+FoxP3−) in the spleens of C57BL/6 mice 24 hours following sham surgery or FPI. Sham + vehicle (N = 6); sham + CAP (N = 5); FPI + vehicle (N = 7 WT); FPI + CAP (N = 6). The symbol * indicates P < 0.05.
FPI-induced changes in components of antigen presentation by peripheral lymphocytes
A hallmark of the transition between innate and adaptive (specific) immunity is successful antigen processing and presentation. Antigen processing requires proteolytic cleavage of invariant chain (CD74) into peptides known as class II invariant peptide (CLIP). CLIP functions as a "placeholder" for antigen specific binding to the peptide-binding groove of MHCII molecules. B cells and γδ T cells are among cells that are well characterized as professional APCs [31],[22]. Therefore, we examined the effects of FPI on CLIP and MHCII from peripheral lymphocytes. We observed a significant decrease in cell surface CLIP on B cells (Figure 2A). Conversely, we observed no corresponding changes in the cell surface levels of MHC II on B cells (Figure 2B). In contrast to CLIP on B cells, we observed no significant differences in the level of CLIP per γδ T cells (Figure 2C), but a significant decrease in MHCII on γδ T cells (Figure 2D). These results suggest possible FPI-induced alterations to molecular machinery involved in antigen processing and presentation.
Differential expression of MHCII and CLIP on B and γδ T cells in response to FPI. (A) The mean fluorescence intensity (MFI), indicative of the level of cell surface expression of CLIP is significantly reduced on the surface of B cells at 24 hours after FPI. This reduction in CLIP likely reflects replacement of CLIP by other peptides. (B) Consistent with this interpretation, no corresponding significant decrease in the MFI of MHCII on the surface of B cells was observed at 24 hours after FPI. (C) In contrast, the MFI of CLIP on the surface of γδ T cells remained unchanged, (D) while the level of MHCII significantly decreased at 24 hours after FPI. Taken together, these data suggest that B cells are a likely candidate as APCs after FPI. Sham + vehicle (N = 6); FPI + vehicle (N = 7). The symbol * indicates P < 0.05.
To test the effect of our peptide on influencing the transition between innate and adaptive immunity, we examined CLIP and MHCII expression on B cells and γδ T cells in mice administered CAP 20 minutes after FPI. We examined these two cell types because they are known to express MHCII and present antigens. We observed no significant differences in CLIP expression on the cell surface of B220+ B cells (Additional file 2: Figure S2) following CAP administration after FPI. These results suggest that of these 2 potential APCs (B cells or γδ T cells), the data are more consistent with B cells functioning as APCs in response to TBI. In contrast, the data are less suggestive of γδ T cells serving as APCs based on reduced MHCII (Figure 2D) and therefore a higher ratio of CLIP to MHCII on γδ T cells.
Absence of FPI-induced lymphocyte expansion in invariant chain deficient mice
Because we observed changes in CLIP expression resulting from FPI, we sought to determine the significance of CLIP involvement in immune modulation by FPI, by using mice deficient in invariant chain, CD74. CD74, which is required for processing and presentation of full-length proteins [18],[15], can serve as a survival factor and can function as a growth promoting receptor. In support of the notion that CD74 is required for lymphocyte expansion following FPI, mice lacking CD74 exhibited no net change in spleen cellularity 24 hours following FPI (Figure 3A). Alternatively, wild type mice exhibit a significant expansion of lymphocytes in the spleen at 24 hrs after FPI (Figure 1A). Furthermore, there were no changes in numbers or activation state of any of the subsets of B or T cells (Figure 3B and C, respectively).
FPI-induced alterations in peripheral lymphocytes require CD74. (A) The number of viable cells in the spleens of CD74Def mice is unchanged at 24 hours following sham surgery or FPI. (B) Similarly, the number of B Cells (B220+) and T cells (CD3+) in the spleens of CD74Def mice are not significantly different at 24 hours after sham surgery or FPI. (C) Analysis of specific populations of T cells also revelaed no significant differences at 24 hours after sham surgery or FPI. For these experiments we used 5 sham surgery mice and 6 FPI treated mice. The symbol * indicates P < 0.05.
CLIP antagonism or invariant chain deficiency modifies the cytokine profile in the CNS following FPI
Our previous work has identified numerous changes in cytokines and chemokines in the CNS following an FPI [32]. Because CAP reduced the number of activated splenocytes following FPI, and activated splenocytes are a potential source of cytokines and chemokines, we sought to determine if CLIP antagonism using CAP modifies the cytokine milieu in the CNS. In order to address this, we screened for multiple cytokines, using a Multiplex cytokine assay system (MCYTOMAG-70 K-PMX). The results showed that CAP treatment significantly reduced IL15, IL13, IL10 and monocyte chemoattractant protein (MCP1), also known as CCL2 (Figure 4).
CAP decreases expression of pro-inflammatory cytokines in the brain following FPI. Multiplex analysis of cytokines in tissue from the ipsilateral cortex of C57BL/6 mice following FPI. Note the significant reductions in IL-13, IL-15, CCL2 (MCP1), and IL-10. The reduction of this subset of chemokines and cytokines is consistent with the reduction in cells capable of producing these inflammatory cytokines. Sham + vehicle (N = 4); sham + CAP (N = 4); FPI + vehicle (N = 4); FPI + CAP (N = 4). The symbol * indicates P < 0.05.
Because the absence of invariant chain in the CD74def mice resulted in no splenocyte activation after FPI, we similarly sought to examine the cytokine and chemokine expression in the brains of these KO mice after FPI. We observed significantly less TBI-induced expression of IFNγ, TNFα, regulated on activation normal T cell expressed and secreted (RANTES; AKA CCL5), macrophage inflammatory protein 1β (MIP1β; AKA CCL4) and IL-13, in the CNS of CD74def mice when compared to C57BL/6 mice (which are the genetic background of these mice) (Figure 5).
CD74 Def mice are resistant to FPI-induced increases in pro-inflammatory cytokines in the brain following TBI. Multiplex analysis of cytokine expression in tissue from the ipsilateral cortex, comparing the cytokine response to FPI in wild type C57BL/6 mice to CD74Def mice. The results demonstrate a significant decrease in the levels of TNFα, IFNγ, RANTES, MIP1β, and IL-13 in tissue from the ipsilateral cortex of CD74Def mice compared to C57BL/6 mice, measured at 24 hours post FPI. The reduction in this group of cytokines in the CD74Def mice, implicate CD74 in their production following FPI. For these experiments we used five C57BL/6 and six CD74Def. The symbol * indicates P < 0.05.
Invariant chain deficiency or CLIP antagonism confers neuroprotection from FPI
To determine the contribution of CD74 to FPI-induced neurodegeneration, we examined FluoroJade C labeling at 3 days after FPI in WT and CD74Def mice. The 3 day time point was selected because we previously demonstrated that this was the peak of neurodegeneration following FPI [26]. The results demonstrate that relative to vehicle treatment (Figure 6A), treatment with CAP significantly reduced the number of FluoroJade C-labeled cells in the ipsilateral cortex (Figure 6B). Similarly, mice deficient for CD74 (Figure 6C) had decreased FluoroJade C-labeling compared to vehicle treatment (Figure 6A). In addition, Analysis of the overall lesion size revealed that both, CAP treated and CD74def mice had a significantly smaller lesion size compared to WT mice (Figure 6D). Quantitation of Fluorojade-staining in the brain following CAP administration and in CD74def mice, demonstrated a significant reduction in degenerating cells (Figure 6E).
CAP treatment after FPI reduces neurodegeneration and lesion size. (A-C) FluoroJade C labeling at 3 days after FPI. In vehicle treated mice (A), significantly more degenerating neurons are observed compared to mice treated with CAP after FPI (B) and CD74def mice (C). (D) Analysis of the size of the lesion at 3 days after FPI revealed that CAP treatment and CD74Defmice had a significantly smaller lesion in the anterior/posterior plane, compared to sham mice. (E) Quantitative analysis of the number of FluorJade C-labeled cells showed significantly less (P < .01 for both) labeled-cells in the CAP treated and CD74Def mice. For this analysis, we used 6 shams, 5 CAP and 5 CD74Defmice. Scale bars in A-C = 50 μm. The symbol * indicates P < 0.01.
This study is the first to demonstrate a putative role for invariant chain CD74 in CNS damage following brain injury. The data support targeting the transition between innate and adaptive immunity in the periphery as a potential therapeutic approach for TBI. Alterations to peripheral immune cell populations include an increase in cellularity in the spleen, comprised of increased B cells, CD3+ T cells, CD4+ T cells, CD8+ T cells, Tregs and γδ T cells. Further, we found that TBI induces a decrease in cell surface CLIP on B220+ B cells, and a decrease in MHCII on activated γδ T cells, suggesting that B cells may be the dominant APC after TBI. We also determined by using CD74Def mice, that these changes were invariant chain dependent.
The fact that CLIP antagonism and CD74Def mice have a reduced lesion size and decreased neurodegeneration after TBI, supports the involvement of CLIP and/or CD74 in mediating damage in the CNS. Both CD74 and its products, including CLIP, are widely accepted as central players in antigen processing and presentation, and are central to the transition between innate and adaptive immunity [12]. The neuroprotection observed in this study supports the involvement of antigen processing and presentation in the exacerbation of brain injury [17],[18],[33],[34]. While it is clear that CD74 is involved in the damage to the CNS following TBI, several different functions have also been attributed to CD74. These include: survival, signal transduction through cell surface CD74, binding to pro-inflammatory macrophage inhibitory factor (MIF), along with its well-established role in antigen processing and presentation [353 day CitationID="CR38">38]. Based on the current findings, it is not possible to exclude any of these functions of CD74 in mediating neurodegeneration after TBI.
To further elucidate the putative role of CD74 in TBI, we engineered a peptide that competitively antagonizes the proteolytic product of CD74, CLIP [25]. We determined that administration of this potentially therapeutic peptide 20 minutes after TBI prevented the increase in splenic cellularity in all observed peripheral immune cell populations with the exception of activated conventional CD4+ T cells. Most importantly, administration of this peptide was neuroprotective following TBI, supporting the notion that neuronal damage involves the transition between innate and adaptive immunity.
The transition between innate and adaptive immunity can be regulated by several subsets of activated, non-conventional T cells, including Tregs and γδ T cells. While Tregs have been well described to dampen antigen-specific responses in the periphery, their role in the CNS is controversial [39]-[41]. Moalem et al. has proposed that following injury to the nervous system, Tregs may be detrimental, because the antigen specific T cell response that they limit may be protective [18],[19],[33]. Alternatively, some investigators have proposed protective effects of Tregs following cerebral insults [40]-[43]. It is pertinent to note that two distinct subsets of Tregs have been characterized, natural Tregs and inducible Tregs [44],[45]. Therefore, it is possible that one population of Tregs is beneficial and the other is detrimental. Future studies are needed to examine this possibility.
The current results also demonstrated a significant expansion of γδ T cells after TBI that could be reversed with CAP treatment. Freedman and colleagues previously demonstrated the presence of γδ T cells in the cerebral spinal fluid of patients with multiple sclerosis [46]. Additionally, it has previously been shown that these γδ T cells could attack oligodendrocytes, indicating the potential for a destructive response by γδ T cells in the CNS [46],[47]. Our data showing a reduced MHCII expression by γδ T cells after FPI argues for reduced capability for antigen presentation and processing. This reduced ability for antigen presentation and processing could prevent an effective transition from innate to adaptive immunity. Such prevention of an effective transition to adaptive immunity could undermine what would otherwise be protective auto-immunity as previously suggested [18],[33].
Similar to γδ T cells, B cells are also well described as antigen specific, capable of producing antibodies, and as exquisitely efficient professional antigen presenting cells [21],[22]. Our finding that after FPI, B cells have significantly reduced CLIP, but not significantly reduced MHCII, suggests that these cells have a higher likelihood for antigen presentation after TBI [22]. In support of this notion, a recent study showed that the antigen presentation function of B cells is critical for the pathology of experimental autoimmune encephalitis (EAE), more so than auto-antibody production [34]. Thus, some consequences of TBI might depend upon efficient antigen processing and presentation by B cells.
The effector functions of B cells and T cells involve cytokines and chemokines. These small molecule compounds serve as chemotactic signaling proteins and immune modulators. Numerous studies have implicated a variety of chemokines and cytokines in the pathogenesis of TBI [32]. The interactions that govern the effects of cytokines are poorly understood in general, and even less-well understood in the context of TBI. In CD74Def mice, we observed significantly less IFNγ and TNFα compared to WT controls following FPI. Elevations of both of these cytokines have been previously reported following various types of TBI [48]-[52], but the cellular source(s) of these cytokines and their functional significance are not well-defined. While some studies demonstrate that glial cells, including microglial cells are involved in the increase [53],[54], the possibility that infiltrating immune cells, including macrophages, contribute to these increases cannot be ruled out. In support of this interpretation, we also observed decreased MIP1β (Figure 5) and RANTES, both of which are products of CD8+ T cells [55],[56], which were also reduced with CAP (Figure 1). Nevertheless, our data with CD74Def mice implicate antigen processing involving CD74 as a contributor to the elevation of these cytokines and chemokines after TBI.
While CCL2 is a well-described marker of inflammation after TBI [57],[58], and CAP reduced its expression in this study, less understood is the putative roles of IL15, IL13 and IL10, that were also reduced by CAP. Interestingly, a previous study using IL15 KO mice and IL15R KO mice in a facial axotomy model, found reduced T cell infiltration and increased MHCII + microglial cells [59]. Consistent with the potential involvement of macrophages/microglial cells, IL13 has been previously shown to be involved in the elimination of activated microglial cells subsequent to an injury [60]. In our study, administration of CAP reduced the lesion size, which could reflect a reduction in the number of activated microglial cells responding to the injury, thereby decreasing the overall level of IL15 and IL-13. With regard to the CAP-dependent decrease in IL-10, previous studies have indicated that elevated IL-10 corresponds with a poor outcome for patients [61],[62]. Therefore, the reduction in IL-10 observed following CAP treatment may be indicative of the decreased lesion size and thus an improved outcome.
The synthesis of CAP was based on the rationale that antagonism of CLIP would selectively target the transition between innate and adaptive immunity. While our results are consistent with this hypothesis, the consequences of CAP therapy resulting in neuroprotection require further examination to fully elucidate the precise mechanisms involved. Nonetheless, the results from the current study provide a foundation for targeting the switch between innate and adaptive immunity for the treatment of TBI. Moreover, our specific characterization of peripheral alterations to distinct subsets of immune cells may provide biomarkers for diagnosing and characterizing the degree of TBI severity, progression and/or recovery.
RPT helped design experiments, performed experiments, analyzed data, and helped write the manuscript. SM helped design and perform experiments. JMK performed cytokine multiplex experiments. SKR, SKH, and HLM performed experiments. MKNR conceived the study, designed experiments, analyzed data, and helped write the manuscript. LAS helped conceive the study, designed experiments, analyzed data, and helped write the manuscript. All authors have read and approved the final manuscript.
RPT is a postdoctoral fellow, Department of Surgery, Texas A&M University College of Medicine. SM is a research scientist, Department of Surgery, Texas A&M University College of Medicine. JMK is a research assistant, Department of Surgery, Central Texas Veterans Health Care System, Texas A&M University College of Medicine. SKR is an undergraduate student, Department of Anthropology, University of Texas, Austin. SKH is a research assistant, Department of Surgery, Baylor Scott and White Health. HLM is a research assistant, Department of Surgery, Texas A&M University College of Medicine. MKNR is a Professor, Department of Surgery, Texas A&M University College of Medicine, and Baylor Scott and White Health. LAS is an Assistant Professor, Department of Surgery, Central Texas Veterans Health Care System, Texas A&M University College of Medicine.
Bazarian JJ, McClung J, Shah MN, Cheng YT, Flesher W, Kraus J: Mild traumatic brain injury in the United States, 1998-2000. Brain Inj 2005, 19(2):85–91. 10.1080/02699050410001720158
Corrigan JD, Selassie AW, Orman JA: The epidemiology of traumatic brain injury. J Head Trauma Rehabil 2010, 25(2):72–80. doi:10.1097/HTR.0b013e3181ccc8b4 10.1097/HTR.0b013e3181ccc8b4
Hyder AA, Wunderlich CA, Puvanachandra P, Gururaj G, Kobusingye OC: The impact of traumatic brain injuries: a global perspective. NeuroRehabilitation 2007, 22(5):341–353.
Beschorner R, Schluesener HJ, Gozalan F, Meyermann R, Schwab JM: Infiltrating CD14+ monocytes and expression of CD14 by activated parenchymal microglia/macrophages contribute to the pool of CD14+ cells in ischemic brain lesions. J Neuroimmunol 2002, 126(1-2):107–115. 10.1016/S0165-5728(02)00046-2
Das M, Mohapatra S, Mohapatra SS: New perspectives on central and peripheral immune responses to acute traumatic brain injury. J Neuroinflammation 2012, 9: 236. doi:10.1186/1742–2094–9-236 10.1186/1742-2094-9-236
Finnie JW: Neuroinflammation: beneficial and detrimental effects after traumatic brain injury. Inflammopharmacology 2013, 21(4):309–320. doi:10.1007/s10787–012–0164–2 10.1007/s10787-012-0164-2
Clausen F, Lorant T, Lewen A, Hillered L: T lymphocyte trafficking: a novel target for neuroprotection in traumatic brain injury. J Neurotrauma 2007, 24(8):1295–1307. doi:10.1089/neu.2006.0258 10.1089/neu.2006.0258
Medawar PB: Immunity to homologous grafted skin; the fate of skin homografts transplanted to the brain, to subcutaneous tissue, and to the anterior chamber of the eye. Br J Exp Pathol 1948, 29(1):58–69.
Butterfield TA, Best TM, Merrick MA: The dual roles of neutrophils and macrophages in inflammation: a critical balance between tissue damage and repair. J Athl Train 2006, 41(4):457–465.
Segel GB, Halterman MW, Lichtman MA: The paradox of the neutrophil's role in tissue injury. J Leukoc Biol 2011, 89(3):359–372. doi:10.1189/jlb.0910538 10.1189/jlb.0910538
Hernandez-Ontiveros DG, Tajiri N, Acosta S, Giunta B, Tan J, Borlongan CV: Microglia activation as a biomarker for traumatic brain injury. Front Neurol 2013, 4: 30. doi:10.3389/fneur.2013.00030 10.3389/fneur.2013.00030
Blum JS, Wearsch PA, Cresswell P: Pathways of antigen processing. Annu Rev Immunol 2013, 31: 443–473. doi:10.1146/annurev-immunol-032712–095910 10.1146/annurev-immunol-032712-095910
Allen PM, Babbitt BP, Unanue ER: T-cell recognition of lysozyme: the biochemical basis of presentation. Immunol Rev 1987, 98: 171–187. 10.1111/j.1600-065X.1987.tb00524.x
Barker CF, Billingham RE: Immunologically privileged sites. Adv Immunol 1977, 25: 1–54. 10.1016/S0065-2776(08)60930-X
Slavin AJ, Soos JM, Stuve O, Patarroyo JC, Weiner HL, Fontana A, Bikoff EK, Zamvil SS: Requirement for endocytic antigen processing and influence of invariant chain and H-2-M deficiencies in CNS autoimmunity. J Clin Invest 2001, 108(8):1133–1139. doi:10.1172/JCI13360 10.1172/JCI13360
Ling C, Sandor M, Suresh M, Fabry Z: Traumatic injury and the presence of antigen differentially contribute to T-cell recruitment in the CNS. J Neurosci 2006, 26(3):731–741. doi:10.1523/JNEUROSCI.3502–05.2006 10.1523/JNEUROSCI.3502-05.2006
Schori H, Lantner F, Shachar I, Schwartz M: Severe immunodeficiency has opposite effects on neuronal survival in glutamate-susceptible and -resistant mice: adverse effect of B cells. J Immunol 2002, 169(6):2861–2865. 10.4049/jimmunol.169.6.2861
Schori H, Shechter R, Shachar I, Schwartz M: Genetic manipulation of CD74 in mouse strains of different backgrounds can result in opposite responses to central nervous system injury. J Immunol 2007, 178(1):163–171. 10.4049/jimmunol.178.1.163
Schori H, Yoles E, Wheeler LA, Raveh T, Kimchi A, Schwartz M: Immune-related mechanisms participating in resistance and susceptibility to glutamate toxicity. Eur J Neurosci 2002, 16(4):557–564. 10.1046/j.1460-9568.2002.02134.x
Babbitt BP, Allen PM, Matsueda G, Haber E, Unanue ER: Binding of immunogenic peptides to Ia histocompatibility molecules. Nature 1985, 317(6035):359–361. 10.1038/317359a0
Cheng L, Cui Y, Shao H, Han G, Zhu L, Huang Y, O'Brien RL, Born WK, Kaplan HJ, Sun D: Mouse gammadelta T cells are capable of expressing MHC class II molecules, and of functioning as antigen-presenting cells. J Neuroimmunol 2008, 203(1):3–11. doi:10.1016/j.jneuroim.2008.06.007 10.1016/j.jneuroim.2008.06.007
Lanzavecchia A: Antigen uptake and accumulation in antigen-specific B cells. Immunol Rev 1987, 99: 39–51. 10.1111/j.1600-065X.1987.tb01171.x
Denzin LK, Cresswell P: HLA-DM induces CLIP dissociation from MHC class II alpha beta dimers and facilitates peptide loading. Cell 1995, 82(1):155–165. 10.1016/0092-8674(95)90061-6
Buus S, Sette A, Colon SM, Jenis DM, Grey HM: Isolation and characterization of antigen-Ia complexes involved in T cell recognition. Cell 1986, 47(6):1071–1077. 10.1016/0092-8674(86)90822-6
Newell MK, Tobin RP, Cabrera JH, Sorensen MB, Huckstep A, Villalobos-Menuey EM, Burnett M, McCrea E, Harvey CP, Buddiga A, Bar-Or A, Freedman MS, Nalbantoglu J, Arbour N, Zamvil SS, Antel JP: TLR-mediated B cell activation results in ectopic CLIP expression that promotes B cell-dependent inflammation. J Leukoc Biol 2010, 88(4):779–789. doi:jlb.0410237 [pii] 10.1189/jlb.0410237 10.1189/jlb.0410237
Mukherjee S, Zeitouni S, Cavarsan CF, Shapiro LA: Increased seizure susceptibility in mice 30 days after fluid percussion injury. Front Neurol 2013, 4: 28. doi:10.3389/fneur.2013.00028 10.3389/fneur.2013.00028
Davidson WF, Parish CR: A procedure for removing red cells and dead cells from lymphoid cell suspensions. J Immunol Methods 1975, 7(2-3):291–300. 10.1016/0022-1759(75)90026-5
Amini M, Nahrevanian H, Farahmand M: Pathogenicity variations of susceptibility and resistance to Leishmania major MRHO/IR/75/ER Strain in BALB/c and C57BL/6 mice. Iran J Parasitol 2008, 3(4):51–59.
Oporto VG, Fuentes R, Borie E, Del Sol M, Orsi IA, Engelke W: Radiographical and clinical evaluation of critical size defects in rabbit calvaria filled with allograft and autograft: a pilot study. Int J Clin Exp Med 2014, 7(7):1669–1675.
Franklin KBJ, Paxinos G: The Mouse Brain in Stereotaxic Coordinates. Academic, San Diego; 1997.
Collins RA, Werling D, Duggan SE, Bland AP, Parsons KR, Howard CJ: Gammadelta T cells present antigen to CD4+ alphabeta T cells. J Leukoc Biol 1998, 63(6):707–714.
Mukherjee S, Katki K, Arisi GM, Foresti ML, Shapiro LA: Early TBI-induced cytokine alterations are similarly detected by two distinct methods of multiplex assay. Front Mol Neurosci 2011, 4: 21. doi:10.3389/fnmol.2011.00021 10.3389/fnmol.2011.00021
Moalem G, Leibowitz-Amit R, Yoles E, Mor F, Cohen IR, Schwartz M: Autoimmune T cells protect neurons from secondary degeneration after central nervous system axotomy. Nat Med 1999, 5(1):49–55. doi:10.1038/4734 10.1038/4734
Molnarfi N, Schulze-Topphoff U, Weber MS, Patarroyo JC, Prod'homme T, Varrin-Doyer M, Shetty A, Linington C, Slavin AJ, Hidalgo J, Jenne DE, Wekerle H, Sobel RA, Bernard CC, Shlomchik MJ, Zamvil SS: MHC class II-dependent B cell APC function is required for induction of CNS autoimmunity independent of myelin-specific antibodies. J Exp Med 2013, 210(13):2921–2937. doi:10.1084/jem.20130699 10.1084/jem.20130699
Leng L, Metz CN, Fang Y, Xu J, Donnelly S, Baugh J, Delohery T, Chen Y, Mitchell RA, Bucala R: MIF signal transduction initiated by binding to CD74. J Exp Med 2003, 197(11):1467–1476. doi:10.1084/jem.20030286 10.1084/jem.20030286
Liao H, Bucala R, Mitchell RA: Adhesion-dependent signaling by macrophage migration inhibitory factor (MIF). J Biol Chem 2003, 278(1):76–81. doi:10.1074/jbc.M208820200 10.1074/jbc.M208820200
Meyer-Siegler KL, Iczkowski KA, Leng L, Bucala R, Vera PL: Inhibition of macrophage migration inhibitory factor or its receptor (CD74) attenuates growth and invasion of DU-145 prostate cancer cells. J Immunol 2006, 177(12):8730–8739. 10.4049/jimmunol.177.12.8730
Shi X, Leng L, Wang T, Wang W, Du X, Li J, McDonald C, Chen Z, Murphy JW, Lolis E, Noble P, Knudson W, Bucala R: CD44 is the signaling component of the macrophage migration inhibitory factor-CD74 receptor complex. Immunity 2006, 25(4):595–606. doi:10.1016/j.immuni.2006.08.020 10.1016/j.immuni.2006.08.020
Bittner S, Ruck T, Schuhmann MK, Herrmann AM, Moha ou Maati H, Bobak N, Gobel K, Langhauser F, Stegner D, Ehling P, Borsotto M, Pape HC, Nieswandt B, Kleinschnitz C, Heurteaux C, Galla HJ, Budde T, Wiendl H, Meuth SG: Endothelial TWIK-related potassium channel-1 (TREK1) regulates immune-cell trafficking into the CNS. Nat Med 2013, 19(9):1161–1165. doi:10.1038/nm.3303 10.1038/nm.3303
Li P, Gan Y, Sun BL, Zhang F, Lu B, Gao Y, Liang W, Thomson AW, Chen J, Hu X: Adoptive regulatory T-cell therapy protects against cerebral ischemia. Ann Neurol 2013, 74(3):458–471. doi:10.1002/ana.23815
Li P, Mao L, Zhou G, Leak RK, Sun BL, Chen J, Hu X: Adoptive regulatory T-cell therapy preserves systemic immune homeostasis after cerebral ischemia. Stroke 2013, 44(12):3509–3515. doi:10.1161/STROKEAHA.113.002637 10.1161/STROKEAHA.113.002637
Liesz A, Zhou W, Na SY, Hammerling GJ, Garbi N, Karcher S, Mracsko E, Backs J, Rivest S, Veltkamp R: Boosting regulatory T cells limits neuroinflammation in permanent cortical stroke. J Neurosci 2013, 33(44):17350–17362. doi:10.1523/JNEUROSCI.4901–12.2013 10.1523/JNEUROSCI.4901-12.2013
Zhou W, Liesz A, Bauer H, Sommer C, Lahrmann B, Valous N, Grabe N, Veltkamp R: Postischemic brain infiltration of leukocyte subpopulations differs among murine permanent and transient focal cerebral ischemia models. Brain Pathol 2013, 23(1):34–44. doi:10.1111/j.1750–3639.2012.00614.x 10.1111/j.1750-3639.2012.00614.x
Haribhai D, Williams JB, Jia S, Nickerson D, Schmitt EG, Edwards B, Ziegelbauer J, Yassai M, Li SH, Relland LM, Wise PM, Chen A, Zheng YQ, Simpson PM, Gorski J, Salzman NH, Hessner MJ, Chatila TA, Williams CB: A requisite role for induced regulatory T cells in tolerance based on expanding antigen receptor diversity. Immunity 2011, 35(1):109–122. doi:10.1016/j.immuni.2011.03.029 10.1016/j.immuni.2011.03.029
Weiss JM, Bilate AM, Gobert M, Ding Y, de Lafaille MA C, Parkhurst CN, Xiong H, Dolpady J, Frey AB, Ruocco MG, Yang Y, Floess S, Huehn J, Oh S, Li MO, Niec RE, Rudensky AY, Dustin ML, Littman DR, Lafaille JJ: Neuropilin 1 is expressed on thymus-derived natural regulatory T cells, but not mucosa-generated induced Foxp3+ T reg cells. J Exp Med 2012, 209(10):1723–1742. S1721. doi:10.1084/jem.20120914 10.1084/jem.20120914
Freedman MS, Ruijs TC, Selin LK, Antel JP: Peripheral blood gamma-delta T cells lyse fresh human brain-derived oligodendrocytes. Ann Neurol 1991, 30(6):794–800. doi:10.1002/ana.410300608 10.1002/ana.410300608
Zeine R, Pon R, Ladiwala U, Antel JP, Filion LG, Freedman MS: Mechanism of gammadelta T cell-induced human oligodendrocyte cytotoxicity: relevance to multiple sclerosis. J Neuroimmunol 1998, 87(1-2):49–61. 10.1016/S0165-5728(98)00047-2
Cho HJ, Sajja VS, Vandevord PJ, Lee YW: Blast induces oxidative stress, inflammation, neuronal loss and subsequent short-term memory impairment in rats. Neuroscience 2013, 253: 9–20. doi:10.1016/j.neuroscience.2013.08.037 10.1016/j.neuroscience.2013.08.037
Fan L, Young PR, Barone FC, Feuerstein GZ, Smith DH, McIntosh TK: Experimental brain injury induces differential expression of tumor necrosis factor-alpha mRNA in the CNS. Brain Res Mol Brain Res 1996, 36(2):287–291. 10.1016/0169-328X(95)00274-V
Holmin S, Hojeberg B: In situ detection of intracerebral cytokine expression after human brain contusion. Neurosci Lett 2004, 369(2):108–114. doi:10.1016/j.neulet.2004.07.044 10.1016/j.neulet.2004.07.044
Morganti-Kossman MC, Lenzlinger PM, Hans V, Stahel P, Csuka E, Ammann E, Stocker R, Trentz O, Kossmann T: Production of cytokines following brain injury: beneficial and deleterious for the damaged tissue. Mol Psychiatry 1997, 2(2):133–136. 10.1038/sj.mp.4000227
Shohami E, Novikov M, Bass R, Yamin A, Gallily R: Closed head injury triggers early production of TNF alpha and IL-6 by brain tissue. J Cereb Blood Flow Metab 1994, 14(4):615–619. doi:10.1038/jcbfm.1994.76 10.1038/jcbfm.1994.76
Cheong CU, Chang CP, Chao CM, Cheng BC, Yang CZ, Chio CC: Etanercept attenuates traumatic brain injury in rats by reducing brain TNF- alpha contents and by stimulating newly formed neurogenesis. Mediat Inflamm 2013, 2013: 620837. doi:10.1155/2013/620837 10.1155/2013/620837
Chio CC, Chang CH, Wang CC, Cheong CU, Chao CM, Cheng BC, Yang CZ, Chang CP: Etanercept attenuates traumatic brain injury in rats by reducing early microglial expression of tumor necrosis factor-alpha. BMC Neurosci 2013, 14: 33. doi:10.1186/1471–2202–14–33 10.1186/1471-2202-14-33
Cocchi F, DeVico AL, Garzino-Demo A, Arya SK, Gallo RC, Lusso P: Identification of RANTES, MIP-1 alpha, and MIP-1 beta as the major HIV-suppressive factors produced by CD8+ T cells. Science 1995, 270(5243):1811–1815. 10.1126/science.270.5243.1811
Song A, Nikolcheva T, Krensky AM: Transcriptional regulation of RANTES expression in T lymphocytes. Immunol Rev 2000, 177: 236–245. 10.1034/j.1600-065X.2000.17610.x
Liu S, Zhang L, Wu Q, Wu Q, Wang T: Chemokine CCL2 induces apoptosis in cortex following traumatic brain injury. J Mol Neurosci 2013, 51(3):1021–1029. doi:10.1007/s12031–013–0091–8 10.1007/s12031-013-0091-8
Semple BD, Bye N, Rancan M, Ziebell JM, Morganti-Kossmann MC: Role of CCL2 (MCP-1) in traumatic brain injury (TBI): evidence from severe TBI patients and CCL2−/− mice. J Cereb Blood Flow Metab 2010, 30(4):769–782. doi:10.1038/jcbfm.2009.262 10.1038/jcbfm.2009.262
Huang Z, Ha GK, Petitto JM: IL-15 and IL-15R alpha gene deletion: effects on T lymphocyte trafficking and the microglial and neuronal responses to facial nerve axotomy. Neurosci Lett 2007, 417(2):160–164. doi:10.1016/j.neulet.2007.01.086 10.1016/j.neulet.2007.01.086
Yang MS, Park EJ, Sohn S, Kwon HJ, Shin WH, Pyo HK, Jin B, Choi KS, Jou I, Joe EH: Interleukin-13 and -4 induce death of activated microglia. Glia 2002, 38(4):273–280. doi:10.1002/glia.10057 10.1002/glia.10057
Ferreira LC, Regner A, Miotto KD, Moura S, Ikuta N, Vargas AE, Chies JA, Simon D: Increased levels of interleukin-6, -8 and -10 are associated with fatal outcome following severe traumatic brain injury. Brain Inj 2014, 28(10):1311–1316. doi:10.3109/02699052.2014.916818 10.3109/02699052.2014.916818
Schneider Soares FM, Menezes de Souza N, Liborio Schwarzbold M, Paim Diaz A, Costa Nunes J, Hohl A, Abreu N, da Silva P, Vieira J, Lisboa de Souza R, More Bertotti M, Schoder Prediger RD, Neves Linhares M, Bafica A, Walz R: Interleukin-10 is an independent biomarker of severe traumatic brain injury prognosis. Neuroimmunomodulation 2012, 19(6):377–385. doi: 10.1159/000342141 10.1159/000342141
This material is the result of work supported by the following: the Scott and White Healthcare Foundation (MKNR), the Wounded Warrior Fund (MKNR), VG Life Sciences Inc. (MKNR, RPT, LAS), Texas A&M Health Sciences Center (MKNR, LAS) and the use of facilities at the Texas A&M Health Science center (MKNR) and the Central Texas Veterans Health Care System, Temple, TX, USA (LAS). We are grateful to Evan W. Newell for his generous contributions to the design and generation of CAP. We gratefully acknowledge Glen Cryer for his support and his editorial assistance with the manuscript.
Department of Surgery, Texas A&M University Health Science Center, Temple, TX, USA
Richard P Tobin
, Sanjib Mukherjee
, Jessica M Kain
, Heather L Motal
, M Karen Newell Rogers
& Lee A Shapiro
Central Texas Veterans Health Care System, Temple, TX, USA
Sanjib Mukherjee
Department of Anthropology, University of Texas, Austin, TX, USA
Susannah K Rogers
Department of Surgery, Baylor Scott and White Health, Temple, TX, USA
Stephanie K Henderson
Texas A&M Health Science Center, Baylor Scott and White Health, 702 SW. HK Dodgen Loop, MRB. Rm. 114B, Temple, 76504, TX, USA
M Karen Newell Rogers
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Correspondence to M Karen Newell Rogers.
RPT, SKR, JMK, and MKNR are scientific consultants for VG Life Sciences Inc., the license holder for CAP. RPT, MKNR, and LAS are all listed on a patent pending for CAP. Patent filed in the United States Patent and Trademark Office on September 9, 2013 under No. 61/875,670, bearing Attorney Docket No. V0223.70010US00.
Additional file 1: Figure S1.: Flow cytometric-gating strategy. Representative 5% contour plots of flow cytometric data to show gating strategy. (TIFF 1 MB)
Additional file 2: Figure S2.: The impact of CAP on the cell surface levels of CLIP and MHCII on B and γδ T cells. (A) The mean fluorescence intensity (MFI) of CLIP on the surface of B cells in the spleens of C57BL/6 mice 24 hours following sham surgery or FPI. (B) The number of CLIP+ B cells (B220+CLIP+). (C) The MFI of MHCII on the surface of B cells in the spleens of C57BL/6 mice 24 hours following sham surgery or FPI. (D) The MFI of CLIP on the surface of γδ T cells in the spleens of C57BL/6 mice 24 hours following sham surgery or FPI. (E) The MFI of MHCII on the surface of γδ T cells in the spleens of C57BL/6 mice 24 hours following sham surgery or FPI. Sham + CAP (N = 5); FPI + vehicle (N = 6). (TIFF 595 KB)
Tobin, R.P., Mukherjee, S., Kain, J.M. et al. Traumatic brain injury causes selective, CD74-dependent peripheral lymphocyte activation that exacerbates neurodegeneration. acta neuropathol commun 2, 143 (2014). https://doi.org/10.1186/s40478-014-0143-5
Fluid percussion injury


text:
October 21, 2014 - New York, New York
ENCORES! CELEBRATES RODGERS AND HAMMERSTEIN AND HART
A Gala Event Starring
Cyrille Aimée, Kate Baldwin, Heidi Blickenstaff, Nick Cordero, Irina Dvorovenko, Chase Finlay, Jared Grimes, Shuler Hensley, Lauren Lovette, Brian Stokes Mitchell, Kelli O'Hara, Laura Osnes, and Paulo Szot
Directed by Warren Carlyle
Musical Direction by Rob Berman
Monday, October 27, 2014 at New York City Center
Encores! Celebrates Rodgers and Hammerstein and Hart, New York City Center's annual Gala on October 27, 2014, will feature the music and lyrics of Richard Rodgers, Oscar Hammerstein II, and Lorenz Hart, sung by some of Broadway's finest talent, including Kate Baldwin, Nick Cordero,Kelli O'Hara, Laura Osnes, Brian Stokes Mitchell and Paulo Szot, with dance performances by Irina Dvorovenko, Chase Finlay, Jared Grimes, and Lauren Lovette. Directed by 2014 Tony winner Warren Carlyle (After Midnight; Hugh Jackman, Back on Broadway), the star-studded show will feature the Encores! Orchestra, led by Encores! Music Director Rob Berman. The Gala evening will honor Bobbie and Lew Frankfort for their passionate, enduring commitment to City Center's education programs, and will help fund New York City Center's artistic and education programs.
The celebration will begin at 6:00 pm with cocktails at City Center, followed by the show at 7:00 p.m. and a post-performance dinner in the Grand Ballroom of The Plaza Hotel (58th Street at 5th Avenue). Gala co-chairs are Perry and Marty Granoff; Elizabeth and Dean Kehler; Victor Luis, Coach, Inc.; Stacey and Eric Mindich; Stuart Vevers, Coach, Inc.; Lisa and Richard Witten; and Barbara and David Zalaznick. The Gala performance is generously underwritten by Hudson Yards: Developed by Related Companies and Oxford Properties Group.
The Gala will feature performances by Cyrille Aimée (A Bed and A Chair at City Center), Kate Baldwin (Big Fish, Finian's Rainbow at Encores!), Heidi Blickenstaff (The Most Happy Fella at Encores!, [title of show]), Nick Cordero (Bullets Over Broadway), Irina Dvorovenko (On Your Toes at Encores!, former principal dancer with the American Ballet Theatre), Chase Finlay (principal dancer with New York City Ballet), 2014 Astaire Award winner Jared Grimes (After Midnight), Tony winner Shuler Hensley (Oklahoma!, The Most Happy Fella at Encores!), Lauren Lovette (soloist with New York City Ballet), Brian Stokes Mitchell (Ragtime, The Band Wagon), Kelli O'Hara (The Bridges of Madison Country, South Pacific), Laura Osnes (Rodgers + Hammerstein's Cinderella, Pipe Dream at Encores!), , and Tony winner Paulo Szot (South Pacific).
New York City Center is thrilled to be honoring Bobbie and Lew Frankfort with the 2014 Fiorello H. LaGuardia Award for distinguished service to New York City and New York City Center. Bobbie Frankfort has been a City Center Board member since 2005, and serves as the Chair of City Center's Education Committee and is a member of the Steering Committee. Bobbie and Lew Frankfort, the Executive Chairman of Coach, Inc., have demonstrated a deep commitment to City Center's education programs, which reach more than 9,000 children each year and help guarantee an arts education to the most underserved kids in the city. The Frankfort's impact on City Center is comprehensive and enduring, and they are most deserving of this honor.
Led by Artistic Director Jack Viertel, New York City Center's Encores! series celebrates the rarely heard works of America's most important composers and lyricists. For each year since its founding in 1994, Encores! has given three glorious Broadway scores the chance to be heard as their creators originally intended. Every production is a whirlwind, with just eight days of rehearsals, one day of dress rehearsal, and seven performances. The cast is backed by the Encores! Orchestra, often more than 30 players strong. The Encores! program received a special 2000 Tony Honor for Excellence in the Theatre, as well as an Outer Critics Circle Award, Lucille Lortel Award, and Jujamcyn Theater Award. Over the years Encores! has presented the works of Harold Arlen, Burt Bacharach, Leonard Bernstein, Jerome Kern, Cole Porter, Stephen Sondheim, and many others. The 2015 Encores! season will include George and Ira Gershwin's Lady, Be Good, Lerner and Loewe's Paint Your Wagon, and John Kander and Fred Ebb's Zorba!
New York City Center (Arlene Shuler, President & CEO) has played a defining role in the cultural life of the city since 1943. It was Manhattan's first performing arts center, dedicated by Mayor Fiorello La Guardia with a mission to make the best in music, theater, and dance accessible to all audiences. Today, City Center is home to many distinguished companies, including Alvin Ailey American Dance Theater, City Center's Principal Dance Company, as well as Manhattan Theatre Club; a roster of renowned national and international visiting artists; and its own critically acclaimed and popular programs. The Tony-honored Encores! musical theater series has been hailed as "one of the very best reasons to be alive in New York." In summer 2013, City Center launched Encores! Off-Center, a new series featuring landmark musicals filtered through the lens of today's most innovative artists. Dance has been integral to the theater's mission from the start, and dance programs, including the annual Fall for Dance Festival, remain central to City Center's identity. Vital partnerships with arts organizations including Jazz at Lincoln Center and London's Sadler's Wells Theatre enhance City Center's programmatic offerings. City Center is dedicated to providing educational opportunities to New York City students and teachers with programs such as Encores! In Schools and the Young People's Dance Series. Special workshops cater to families, seniors and other groups, while events such as the Fall for Dance DanceTalk series offer learning opportunities to the general public. In October 2011, City Center completed an extensive renovation project to revitalize and modernize its historic theater.
Tickets for the Gala show only are $30 – $115 and are available online at www.NYCityCenter.org, at the box office (West 55th Street between 6th and 7th Avenues) or by calling CityTix at 212.581.1212.
Tickets for the Gala show, cocktails and dinner are $500 – $10,000 per person. Gala table and ticket details are available by calling 212.763.1205. For performance-only tickets, call CityTix at 212-581-1212 or visit www.NYCityCenter.org.


Some have claimed that Solove was merely a "random passerby" in Charleston whose views weren't important to the larger story, but an investigation by Breitbart News shows that while Solove may be outspoken, she's a recognized and respected activist in the modern Black Power movement.
For those without faith, Michael Wear acknowledges, this may make little sense, especially for those who have never personally experienced the forgiveness of God. "But that does not grant us the right to whitewash the motivation for the forgiveness we witnessed in Charleston," he said. We need to "take the family members seriously when they say it is a sincere, thought-out expression of their faith," he wrote.
There is great power in symbols, especially in an era when so many people have been taught to emote rather than thinking. The quick surge of emotion people feel upon viewing a potent symbol can be bottled and used in politics. It's excessively charitable to call the current flag battle a "debate," because it mostly consists of one side screaming that anyone who hesitates to dump that flag into the wood chipper of historical oblivion is either a racist, or an accessory to racism in the first degree. There isn't much in the way of measured reason involved here.
On June 23, Sacramento Flag Works ceased sales of the the Confederate flag because Dylann Roof allegedly shot and killed nine people at Charleston, South Carolina's Emanuel African Methodist Episcopal Church last Wednesday.
Haley left a door ajar that the left intends to prop wide open, from now until Election Day and beyond. Emboldened by South Carolina's quick shift—on an issue that was tangential to the horrific atrocity in Charleston—the left is eager to mount a continued campaign against the Confederacy and its symbols.
Rapper Lupe Fiasco penned a lengthy three-part open letter addressed to "White Supremacy" over the weekend in response to the horrific killings of nine people at a black church in Charleston, South Carolina last week.
Late-night comic and South Carolina native Stephen Colbert joined thousands of marchers in Charleston on Sunday to form a "unity chain" in response to the murders of nine people at a black church last week.
Over the weekend, leftist politicians and the media blamed the racist terror attack in Charleston, South Carolina on conservatives.
The dead are not yet buried in Charleston, yet the South Carolina legislature is to meet in special session Tuesday to debate the removal of the Confederate battle flag from the Capitol grounds, reportedly at the behest of Gov. Nikki Haley. Whether the flag belongs there or not–and I would argue that it does not–the flag had nothing to do with the nine murders in a church last week. To tear it down in such haste is to dishonor the dead–and to accept a collective guilt that knows no end.
On Friday, a Democrat South Carolina state legislator appeared on CNN insisting that the 21-year-old man who killed nine churchgoers in Charleston last week was driven to his act because he watches Fox News. This is despite the fact that no one seems to know what media habits the killer possessed.
On June 18, President Obama called for Hollywood celebrities to join him in pushing gun control in the wake of the Charleston attack but not every celebrity is falling in line.
During an interview with people gathered outside the Mother Emmanuel American Methodist Church in Charleston, South Carolina, where Wednesday's deadly shooting left nine black churchgoers dead, a woman called for a "race war" and discussed black anger.
On Friday evening, July 2, 1999, a crazed young man drove through my Chicago neighborhood looking for Jews to kill. He had already shot half a dozen in a neighboring area (all of whom survived). He saw a black man walking with his family and decided that was good enough. He shot him–and Ricky Byrdsong, former Northwestern University basketball coach, died. The killer led an interstate manhunt, during which he killed a Korean man and attacked a church before shooting himself.
President Obama wasted no time in light of the massacre at the historic and symbolic Emanuel African Methodist Episcopal Church in downtown Charleston, South Carolina.
The killer evidently attended the prayer meeting he attacked for nearly an hour before opening fire and killing nine people.


The living room fills with the tiny click-clicks of a keyboard. I haven't bothered to put my ear buds in or even glance at the lovely stack of books on the coffee table that have tried to tempt me with their gleaming, paperback covers. Vanilla Turkish Taffy twists like a sticky ribbon in my fingers as I fiddle with it and squint at the text on my laptop screen.
If my sentences could dance, they'd jerk and jive like someone threw ice down their backs. "Too awkward," I mumble. I separate paragraphs and stare each sentence down. I make a list on my miniature legal pad of what, exactly, I'm trying to accomplish by writing this letter. There's an art to letter-writing, I think. And I'm not sure I've mastered it.
I won't compare letters to telephone calls or even emails. I won't pretend that I, like many people in Jane Austen's time, can read or even write a criss-cross letter. But there's something about composing a letter that will never change: the privacy of it. And nothing says "microcosm" than opening up a page full of handwriting. But here's a confession. I haven't handwritten a letter in a long time. I actually tried to do it recently, but my handwriting isn't that great and, well, I write a lot. I'm one of those rambling writers. I feel like I have too much to say, and I get excited, and, being a writer, I feel that if I have just one more paragraph, I can make you feel as if you were right there with me. Or that's what I'd like to do one day.
So if you receive a letter from me that's only a page long, you'll know I've labored over it, haha. Whether I'm handwriting a letter or typing it, I often feel like the air around me disappears. I try to imagine the person I'm writing to. Would my words make her smile? Can that joke sound funny without a "just kidding" directly after? Will he understand what I'm saying when I try to relate?
This is why people are paid to write greeting cards, right? When I browse the card aisles, I can't help but admire even the cheesiest of lines. The point comes across. The message is clear. This is why people will buy expensive cards with golden, swirly words and only sign their names on the inside.
No matter how much trouble a letter can be, it's one of the best feelings in the world to send it on its way. I feel warm and happy, knowing that my letter is changing hands, hitching rides in people's pockets, flying overhead, or sailing across the sea. And very soon, the seal will be broken and the letter unfolded. There's magic in that.


You're in the thick of it!
When you arrive you'll be given a new identity to protect the innocent.
Use your finely honed deductive reasoning to solve the mystery... or you can just bribe people.
At the end of the night one keen detective from the audience will take home the prize.
Prices and menu may vary.
The Mystery Mansion is perfect for Special Occasions. Contact us to book groups of 10 or more.
Whether it's an Anniversary, Birthday, or Cult "Returning Home" Party, Mystery Mansion makes any events unforgettable.
Bring Mystery Mansion to your customers. Businesses looking to host a special event or boost a slow night will benefit from our Event Packages.


We receive many emails and letters asking me about chemtrails. Many express deep concern, some are abusive and threatening. This page gives our views on the topic.
Chemtrails refers to the theory that governments or other parties are engaged in a secret program to add toxic chemicals to the atmosphere from aircraft in a way that forms visible plumes in the sky, somewhat similar to contrails. Various different motivations for this alleged spraying are speculated, including sterilization, reduction of life expectancy, mind control or weather control.
We have not seen any credible evidence that chemtrails exist. If we did see any evidence that governments were endangering their own citizens in the manner alleged in the chemtrails conspiracy, we would be eager to expose and stop any such activities.
We work on theoretical proposals for combating global warming called solar geoengineering or albedo modification. These are technologies that might enable people to add materials to the Earth's atmosphere to reflect a bit more sunlight back to space, partially masking or reducing the climate change due to accumulated greenhouse gases.
Study of solar geoengineering is in the very early stages and the topic is (rightly) a very controversial area of climate policy because if it ever were tested at large scales or implemented it could involve physical risks and would raise a range of serious socio-political and ethical issues. However, there is nothing secret about the study of albedo modification, with hundreds of publications about the topic in academic journals and hundreds of articles in mainstream media. You can find some of our work here.
We are confident that there is no currently active program to actually test or implement albedo modification outdoors.
We are not now involved in outdoor experimentation, though we are indeed actively developing proposals for field experiments. This experiment will proceed only if it is conducted in a fully transparent and public manner, and only if it passes a comprehensive independent safety review. The experimental plans, operations and results will be publicly available and freely usable (no patenting).
Because of the apparent similarities between the proposed implementation methods for albedo modification (such as injecting reflective particles into the stratosphere), and the alleged methods for producing chemtrails, some people have linked the notion of chemtrails to the study of albedo modification.
Extraordinary claims require extraordinary proof. The claim that there is a large-scale secret program to spray materials from aircraft is extraordinary. Yet all the evidence we have seen to date has been very weak. The most common claim is simply that aircraft contrails look "different", without any comparative analysis. This as convincing as saying that alien beings walk among in disguise as people because some people act very strangely.
If there really were a large-scale program dumping material from aircraft at the scale described, there would have to be a large operating program to manufacture, load and disperse materials. If such a program existed at the scale required to explain the claimed amount of chemtrails, it would require thousands or perhaps tens of thousands of people. It would be extraordinarily hard to keep such a program secret because it would be so easy for a single individual in the program to reveal it using leaked documents, photographs or actual hardware.
Moreover, if such a program was intended to cause harm to their fellow citizens—as is alleged by people who believe in the chemtrails conspiracy—then people working in the program would have very strong personal motivations to reveal it.
We don't trust that governments will always do the right thing, but we do trust the goodwill of individuals who take risks to blow the whistle, exposing government misdeeds.
If you believe in chemtrails, ask yourself how you can be so certain that there is indeed a grand conspiracy. Consider alternative, simple explanations. Remember that the Internet is filled with people who are completely sure about stuff that just isn't true. Remember that while governments can keep secrets that involve only a few people they do a very bad job of keeping large-scale programs secret. The Russians, for example, knew specific details about the Manhattan project before the first nuclear weapons test, and that was a case where those involved had a clear patriotic motivation to keep the secret.
If you think that aircraft contrails "look different" or are "behaving strangely" consider that regular aircraft condensation trails (contrails) are sometimes long-lasting and the way they look can change suddenly along a flight path as aircraft fly through regions with different temperature and humidity. This is well understood and has been demonstrated by ground observation and by scientific research performed by thousands of people over many decades. Of course it's possible that something else is going on, but to prove it one would need more than simple photographs of something that looks entirely consistent with normal contrails.
Several people who were concerned about chemtrails came to talk with me and recorded a video of our conversation.


Our current internal and external job openings are listed below. Please click on a posting to open or save it. If you would like to apply to a posted vacancy please forward your resume and cover letter clearly indicating the position you are interested in.
Internal postings are intended for members of the bargaining unit (members of CUPE Local 3393).
If there are no posted vacancies you are still welcome to submit your resume. We will keep it on file for one (1) year. Resumes can be sent to jobs@westnh.org.
Please visit our website frequently to view our current job postings and re-submit your cover letter and resume as specified in the posting to be considered for the vacancy.
Thank you for your interest in working for West Neighbourhood House!


If you're doing well and you've found value in these tools, I'm asking for your financial support. I'll keep creating content, building HeroMuster, and trying to make the world a better place for roleplayers. No hard feelings if you can't help.
Click the donate button to donate through PayPal (or if you have a HeroMuster account, login and buy coins to use a credit card).
Every reload creates a new, procedurally generated NPC.
You gain advantage 7 on lethal strikes. Up to 20 damage (not to exceed the total damage dealt) is considered lethal damage. Your attacks are considered lethal strikes whenever you fulfill one of the following conditions: Your target is caught off guard or otherwise unaware of the attack, such as when you are hidden from them, disguised as a friend, or have successfully deceived them; Your target is within melee attack range of an ally.
Legendary creature names and descriptions sourced from Wikipedia.


text:
March 25, 2012, Volume 2 # 15
Next Lecture Tuesday April 17, 2012
The next lecturer will be Lila Levinson who will talk about her new book, Gated Grief, the story of her father, an army surgeon who landed at Utah Beach on June 6, 1944 along with the first wave of American Soldiers. As usual the lecture will commence at 7 p.m. at the Millburn Library. Levinson's father tended to hundreds of wounded soldiers but was not prepared for what he experienced, when among the first group of Americans who liberated Nordhausen, the infamous Nazi Concentration Camp, he saw what happened to tens of tens of thousands of Jewish prisoners who had perished. The experience so affected him that he managed to virtually suppress the horrible memories and never discussed what he had seen his entire life. However, he did maintain careful records and pictures which he stored in a locked footlocker. His daughter discovered these records and documents after her father's death and decided the story was so important that she had to tell what her father was loath to discuss. Gated Grief is the result of years of research and countless interviews with war veterans and their families.
Lila is a resident of Texas and the Book Club is fortunate that Lila will travel to New Jersey to tell her father's story. Lila taught a Holocaust literature course at St. Edward's University in Austin Texas. She is a freelance commentator on Cultural issues and her work has appeared in numerous newspapers and periodicals. The Writers League of America named Gated Grief the best narrative nonfiction book of the year. Levinson's book will be available for purchase and signing after the lecture.
Andrew Nagorski
Andrew's lecture March 20 about his new book Hitlerland was a huge success. About 80 people were in attendance and thoroughly enjoyed his presentation. Twenty five books were sold and autographed. This was Andrew's second appearance at the Book Club.
On Tuesday May 15, 2012 Dr. Josh Botts, Ph.D. from the State Department will give a talk on the most important, and clearly the most controversial Military Conference of World War II, the Yalta Conference. President Roosevelt was a sick man in February 1945 when he attended the Yalta Conference, dying less than two months later on April 12. Historians have debated for years whether Roosevelt, because of his health, and his determination to stake so much on the creation of the United Nations, gave Stalin too much and in the process set the stage for the Cold War. This will be an exciting and informative lecture.
The Wedemeyer Biography
John J. McLaughlin's biography of General Albert C. Wedemeyer is due for release this May. The name of the book is General Albert C. Wedemeyer, The Unsung Strategist of World War II. John will give a talk about his book on Tuesday, June 19, at 7 p.m.
There are no lectures during the months of July and August. The Fall schedule will be posted in the next Newsletter.
Andrew Nagorski's March 20 Lecture
Andrew Nagorski's March 20 presentation of his latest book, Hitlerland, was quite simply a smashing success, as evidenced both by the large number of members in the audience and the lively question and answer period that followed. As described by Nagorski, most contemporaneous accountings of Hitler tragically underestimated his drive to power. Pictures from the evening have been posted to the Club's Flickr gallery, as always, thanks to Sharon Austin. View the pictures from March 20 here >>


text:
American Rankings
PABCON Championships
2016 Cali, COL
2015 San Juan, PRC
2014 Colombia
2009 Puerto Rico
2008 Chile
2007 Guatemala
PABCON Senior
PABCON CoC
PABCON Youth
Pan Am Games
CONCABA
CONCECA
South American C'ships
Central American Games
Ibero-American C'ships
Macpherson, Dorin-Ballard ready to represent Team USA at PABCON Women's Championships
Happyland Bowling Center Santiago de Chile (Oct. 19-24, 2008)
Over the course of their careers, Wendy Macpherson of Henderson, Nev., and Carolyn Dorin-Ballard of North Richland Hills, Texas, have experienced just about every aspect of competitive bowling. Now, they are about to take the next step.
For the first time, Macpherson and Dorin-Ballard will compete as members of Team USA when they lead a group of the best female bowlers in the country to the 2008 Pan American Bowling Confederation Women's Championships in Santiago, Chile, from Oct. 20-24.
The two Team USA newcomers will be joined by Diandra Asbaty of Chicago, Lynda Barnes of Double Oak, Texas, Liz Johnson of Cheektowaga, N.Y., and Kelly Kulick of Union, N.J., all of whom have been a part of the Team USA program in the past.
"It's going to be a real thrill to compete for Team USA, and I couldn't have asked for better teammates," said Macpherson (right), a long-time professional bowler who won 20 titles while on the Professional Women's Bowling Association Tour. "It will be a thrill to bowl with them instead of against them for once."
Players like Macpherson, Dorin-Ballard, Johnson and Kulick, who are professional bowlers, now are eligible to compete with Team USA in international competition, thanks to a change in World Tenpin Bowling Association legislation last year.
"Competing for Team USA is definitely going to be one of the highlights of my career, and it's almost like another chapter," said Dorin-Ballard, a 20-time PWBA champion.
"It's going to be a great new experience. I'm looking forward to being part of a great team and bringing home gold medals for my country."
Competition at the PABCON Women's Championships begins Monday with singles and continues with doubles, trios, team and Masters. Medals also will be awarded in all-events.