text: Home Gallery Print (image) Plastic Pollution Coalition Plastic is Forever - Wetsuit Plastic Pollution Coalition: Plastic is Forever - Wetsuit Reality hits iconic Vancouver sculptures. Published on May 10, 2010 Editor's Pick Rethink brought a dose of reality to some iconic Vancouver sculptures to illustrate the impact of single-use plastic on marine and wildlife, for the Plastic Pollution Coalition. Rate this Ad You must be registered to rate the ad. Please Login or Register Now About CreditsDateMay 10, 2010Agency:Rethink CommunicationsClient:Plastic Pollution CoalitionCreative Director:Chris StaplesCreative Director:Ian GraisArt Director:Leia RogersCopywriter:Leia RogersPhotographer:David de HaasProducer:Sue WilkinsonProducer:Rhonda Merakian CategoryOOH Latest Creativity News on Ad Age Jennifer Aniston Hates Dry Eyes, Hearts EyeLove in New Spot Goodby, Silverstein & Partners Names Margaret Johnson Chief Creative Officer, Derek Robson President How Advertisers Are Tapping Into China's Crazy Live-Streaming Culture Advertisement Ad Age Events Nov 02 Los Angeles, CA SummitAd Age Brand Summit The Ad Age Brand Summit in Los Angeles peels apart the layers of brand building with a carefully crafted roster of top marketing, technology, and creative leaders. Learn more Related work Plastic Pollution Coalition: Plastic is Forever - Porpoises Rethink Communications Plastic Pollution Coalition: Plastic is Forever - Wildlife Rethink Communications Molson: The Beer Fridge Project Indonesia Rethink Communications Want to see your work in our gallery? Submit now


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Ashley Olsen and Lance Armstrong dated from October to December, 2007.
Ashley Olsen is a 30 year old American Actress. Born Ashley Fuller Olsen on 13th June, 1986 in Sherman Oaks, CA,USA and educated at Campbell Hall Episcopal Day School, Hollywood, CA (2004), she is famous for Full House in a career that spans 1987–present. Her zodiac sign is Gemini.
Lance Armstrong is a 44 year old American Cycler. Born Lance Edward Armstrong on 18th September, 1971 in Plano, Texas, USA and educated at Plano East Senior High, Plano, Texas, USA, he is famous for 7-time Tour De France winner. His zodiac sign is Virgo.
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ashely hes to old for you and i know you can do ALOT better!!!!!!!!!!!!!!

subset: commoncrawl


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Get off Lyrics by Betty Blowtorch
Get off by Betty Blowtorch
i can hear you coming
down the hallway
i can feel your breath
on my neck
mom help me
dad he's raping me
dad get off of me
dad you're huring me
i hate you
f**k you
i hate you
f**k you
I was only nine
get off me now
f**k you, you f**king jerk
dad get off
dad stop
f**k you
you're sick
im not to blame
your'e the one who's insane
eat my sh*t
you're a f**king dick
i hate you
f**k you
i hate you
f**k you
my name is talking tiny and i hate you
my name is talking tiny and i am going to kill you
ha ha ha, you're going to die
i can hear you coming
down the hallway
i can feel your breath
on my neck
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Richard Lucas 10 3B
Binghamton Mets
- Double-A | Eastern League
- MLB Parent Club: New York Mets
- Status: Inactive
- Full Name: Richard Henry Lucas
- Age: 27 (November 2, 1988)
- Birthplace: Jacksonville, FL
- Bats/Throws: R/R Ht: 6' 1" Wt: 205
- Draft: Round 4 (2007, NYM)
- School: Samuel W. Wolfson, Jacksonville, FL
Richard Lucas Career Stats
MiLB.com News
- Moncada, Meadows headline AFL rosters
- Chapman, De Jong take top Texas League honors
- Top 2016 Draft pick Moniak joins podcast
- A's bump top prospect Barreto to Triple-A
- Roundup: Phils' Alfaro, O's Sisco stay hot
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|November 4, 2013
|3B Richard Lucas elected free agency.
|April 4, 2013
|3B Richard Lucas assigned to Binghamton Mets from St. Lucie Mets.
|March 26, 2013
|1B Richard Lucas assigned to New York Mets.
|April 5, 2012
|Richard Lucas assigned to St. Lucie Mets from Brooklyn Cyclones.
|April 2, 2012
|3B Richard Lucas assigned to New York Mets.
|June 20, 2011
|Richard Lucas assigned to Brooklyn Cyclones from St. Lucie Mets.
|June 17, 2011
|St. Lucie Mets sent 3B Richard Lucas on a rehab assignment to Brooklyn Cyclones.
|April 7, 2011
|Richard Lucas roster status changed by St. Lucie Mets.
|April 8, 2010
|3B Richard Lucas assigned to St. Lucie Mets from Brooklyn Cyclones.
|August 19, 2009
|Richard Lucas assigned to Brooklyn Cyclones from Kingsport Mets.
|July 17, 2009
|Richard Lucas assigned to Kingsport Mets from GCL Mets.
|July 11, 2009
|Richard Lucas assigned to GCL Mets from Kingsport Mets.
|June 17, 2009
|Kingsport Mets reassigned 3B Richard Lucas to the minor leagues.


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Today's Devotional
Staying TrueOur ability to stand in the midst of trials will depend on the focus of our gaze.
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PIDF Climate Change Adviser Appointed To UN Intergovernmental ...… United Nations Intergovernmental Panel on Climate Change (IPCC). This follows the appointment … Pacific Islands Development Forum (PIDF) climate change adviser Dr Mahendra Kumar to … report on the impacts of global warming of 1.5C above ... EnvironmentGuru.com, 1 month ago
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The North Face Rolling Thunder
|7 lb / 3175 g
|10 lb 9 oz / 4790 g
|11 lb 10 oz / 5275 g
|1955 cu in / 32 L
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|Phthalate-Free TPE Fabric Laminate, 840D Jr Ballistics Nylon
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|Phthalate-Free TPE Fabric Laminate, 840D Jr Ballistics Nylon
Where to Buy
Manufacturer's Description
Visit The North Face's Rolling Thunder page.
Retailers' Descriptions
Here's what other sites are saying:
Strong and durable, the North Face Rolling Thunder is a bombproof gear hauler that is perfect for a week or two abroad. Sleek updated design with a weight reduction of 25%; no compromise to durability or functionality. . Bomber base camp body material paired with rugged nylon for extra durability inside and out. Separate zippered external dirty clothes or shoes compartment. Sherpa strap to carry your backpack or duffel securely on the face of your pack. Perforated exposed back panel reduces weight without sacrificing strength or comfort. Plastic bumper wear points for extra abrasion resistance. Two external zipper pockets on face of bag for additional organization options. Angled handles for easy-wheeling and better weight distribution. Internal split mesh pocket. Die-cut ID window. Coated daisy chains for durable lash points. Extendable luggage handle with patented side release buttons.
Ready for a week or two abroad on adventure travel excursions, this rugged 30 rolling suitcase now sports a lighter, more user-friendly design with an external compartment to stash shoes or dirty clothes.
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When the thunder rolls you'll have to walk faster, but luckily the wheels on The North Face Rolling Thunder Duffel will keep up as you avoid the lightning.
- Backcountry.com
The North Face Rolling Thunder
currently retails for:
$238.90 - $319.00
$238.90 - $319.00


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Pinterest makes its pitch to retailer advertisers
The social network argues that Pinterest is a 'platform of intent' where consumers are finding ideas for things to buy and do.
Pinterest makes its pitch to retailer advertisers
The social network argues that Pinterest is a 'platform of intent' where consumers are finding ideas for things to buy and do.
An investor in China's Xiaomi talks about e-commerce trends
GGV Capital's Hans Tung, who has witnessed highs and lows in internet business financing, talks about recent investments, including a cross-border e-commerce app and ...
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Sponsored by 2013 Social Media 300
Rather than commit considerable resources to paid search, CEO Ryan DeLuca says BodyBuilding.com will likely generate $48 million in web sales in 2005 using word-of-mouth marketing and other less elaborate programs.
Though it's rare that a web retailer isn't devoting at least a portion or, in some cases, all of its advertising budget to search engine marketing, BodyBuilding.com is doing just fine with organic search and other less elaborate marketing tactics.
In fact, BodyBuilding.com, No. 160 in the Internet Retailer Top 400 Guide to Retail Web Sites, expects web sales to reach $48 million in 2005 compared with online sales of $32 million in 2004.
Each day, BodyBuilding.com attracts about 160,000 unique visitors to its site, says CEO Ryan DeLuca. What's driving traffic–and sales–is an online library of nearly 10,000 articles on health, nutrition, weight training, bodybuilding and other topics. "So far we haven't done any paid search," says DeLuca. "We have almost 400 writers, many of whom are experts on their topic, contributing content to the site. We have almost 16,000 pages in the library and adding more all the time. We get good word-of-mouth advertising."
DeLuca bought the rights to BodyBuilding.com for $20,000 several years ago and turned an interest in both bodybuilding and web development into a full-time e-commerce operation. To market the site, DeLuca depends on organic search and some e-mail marketing.
But the site mainly draws traffic from bodybuilders and others who come to BodyBuilding.com to read articles in the library and then stick around to shop the web store. BodyBuilding.com maintains separate portals for the web store and the library, but depends on products mentioned or reviewed in various articles to spark a reader's interest in shopping online.
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In the future, BodyBuilding.com, which fills about 1,850 orders per day at two distribution centers in Idaho and Florida, will add features such as an online tool for maintaining a training log and a vendor extranet. "They can use the extranet to send us updated product information or download feedback on how their products are selling," DeLuca says.
But with a loyal following, BodyBuilding.com will continue to use word-of-mouth advertising rather than paid search to drive traffic. "The site is built on the loyalty we have with readers and shoppers," he says. "We aren't going to change that."
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Posted 08/25/16
Online retail is hard, grocery retail is really hard, so online grocery is, of course, really, really hard."


text:
J. Neurochem. (2010) 114, 1840–1851.
Chronic restraint stress impairs neurogenesis and hippocampus-dependent fear memory in mice: possible involvement of a brain-specific transcription factor Npas4
Address correspondence and reprint requests to Dr Kiyofumi Yamada, Department of Neuropsychopharmacology and Hospital Pharmacy, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8560, Japan. E-mail: email@example.com
Neurogenesis in the hippocampus occurs throughout life in a wide range of species and could be associated with hippocampus-dependent learning and memory. Stress is well established to seriously perturb physiological/psychological homeostasis and affect hippocampal function. In the present study, to investigate the effect of chronic restraint stress in early life on hippocampal neurogenesis and hippocampus-dependent memory, 3-week-old mice were subjected to restraint stress 6 days a week for 4 weeks. The chronic restraint stress significantly decreased the hippocampal volume by 6.3% and impaired hippocampal neurogenesis as indicated by the reduced number of Ki67-, 5-bromo-2′-deoxyuridine- and doublecortin-positive cells in the dentate gyrus. The chronic restraint stress severely impaired hippocampus-dependent contextual fear memory without affecting hippocampus-independent fear memory. The expression level of brain-specific transcription factor neuronal PAS domain protein 4 (Npas4) mRNA in the hippocampus was down-regulated by the restraint stress or by acute corticosterone treatment. Npas4 immunoreactivity was detected in progenitors, immature and mature neurons of the dentate gyrus in control and stressed mice. Our findings suggest that the chronic restraint stress decreases hippocampal neurogenesis, leading to an impairment of hippocampus-dependent fear memory in mice. Corticosterone-induced down-regulation of Npas4 expression may play a role in stress-induced impairment of hippocampal function.
brain-derived neurotrophic factor
- BrdU
dentate gyrus
granule cell layer
glial fibrillary acidic protein
- NeuN
neuronal nuclei
- Npas4
neuronal PAS domain protein 4
phosphate-buffered saline
subgranular zone
- Sox-2
SRY-related HMG box 2
Stress is defined in biological systems as any condition that seriously perturbs physiological/psychological homeostasis and well known to affect the function and morphology of the hippocampus (Kim and Diamond 2002). The exact underlying cellular mechanisms that mediate the inhibitory effect of stress are largely unknown. However, stress reduces the expression of several growth factors and neurotrophic factors, such as brain-derived neurotrophic factor (BDNF), insulin-like growth factor-1, nerve growth factor, epidermal growth factor, and vascular endothelial growth factor, that can all influence neurogenesis (Lucassen et al. 2010).
A variety of stress paradigms also impair hippocampal neurogenesis (Mirescu and Gould 2006). Neurogenesis in the hippocampus occurs throughout life in a wide range of species (Kempermann et al. 1997; Eriksson et al. 1998). The newly divided cells in the subgranular zone (SGZ) of the dentate gyrus (DG) in the hippocampus migrate into the granule cell layer (GCL) of the DG and are integrated into the existing circuitry (Tashiro et al. 2006). Impairment of hippocampal neurogenesis could be one of the etiological factors for neuropsychiatric disorders (Reif et al. 2006), and hippocampal neurogenesis may be a target for the treatment of depression (Airan et al. 2007).
Recently, we have demonstrated that rearing in long-term social isolation after weaning impaired hippocampal neurogenesis and spatial memory in Morris water maze, and reduced expression levels of neuronal PAS domain protein 4 (Npas4) mRNA in the DG of the hippocampus (Ibi et al. 2008). Npas4 is a brain-specific transcription factor and may have a neuroprotective function (Ooe et al. 2009). It has been reported that Npas4 regulates the expression of drebrin, which engages in dendritic-cytoskeleton modulation at synapses in the hippocampus (Ooe et al. 2004). Recently, Npas4 has been shown to control GABAergic synapse development through the transcriptional regulation of BDNF (Lin et al. 2008). However, it remains to be determined whether Npas4 gene expression in the hippocampus is affected by deleterious stress. In this study, we investigated the effect of chronic restraint stress in early life on hippocampal neurogenesis, hippocampus-dependent memory and Npas4 expression in mice.
Materials and methods
Male ICR mice (3 weeks old) were purchased from Japan SLC Inc. (Hamamatsu, Japan). They were housed 5–6 per cage under standard conditions (23 ± 1°C, 50 ± 5% humidity), with a 12-h light/dark cycle. Food and water were available ad libitum. The animals were handled in accordance with the guidelines established by the Institutional Animal Care and Use Committee of Nagoya University.
Restraint stress procedure
Mice were randomly divided into two groups, restraint stress and control groups. Chronic stress was applied for 4 weeks with a stainless mesh that allowed for a close fit to mice (6 h/day between 10 am and 4 pm, 6 days a week) as described previously (Takuma et al. 2007). For acute restraint stress, mice were subjected to the stress for 2 h or 6 h.
Corticosterone level measurement
Corticosterone levels were measured with an enzymatic immunoassay kit (Cayman, Ann Arbor, MI, USA) following the manufacturer's manual with modifications. Briefly, mice were decapitated and 1 mL of blood was collected with 30 μL of 100 mM EDTA immediately after the final restraint stress. Samples were centrifuged (15 min, 1000 g, 4°C). Each supernatant was added to a 96-well plate at two dilutions (1 : 150 and 1 : 300) in duplicate and subjected to the immunoassay. The optical density of the enzyme products was read at 405 nm.
Conditioned-fear test
The conditioned-fear test was carried out as described previously (Nagai et al. 2003). One day after the chronic stress, freezing response was measured in a neutral cage (30 cm × 30 cm × 35 cm high) for 1 min without sound (pre-conditioning phase). In the conditioning phase, each mouse was placed in a training cage (25 cm × 30 cm × 11 cm high) and allowed to explore freely for 2 min, and then a 15-s tone (85 dB) was delivered (conditioned stimulus). During the last 5 s of the tone stimulus, a foot shock of 0.8 mA was presented as an unconditioned stimulus through a shock generator (four times with 15-s intervals.). Context-dependent test was carried out 24 h after the conditioning and the tone-dependent test was conducted 3 h after the context-dependent test. For the context-dependent test, freezing behavior was measured in the training cage for 2 min without tone or foot shock presentation. For the tone-dependent test, the freezing behavior was measured in the neutral cage for 1 min in the presence of a continuous-tone stimulus. After the test phase, the sensitivity to the foot shock was determined. The foot shock was gradually increased in 0.05 mA increments until vocalization.
Nissl staining
Coronal sections (10 μm) through the entire extent of the hippocampus were cut and every fifteenth section was collected. The sections were dehydrated through an ascending series of ethanol concentrations. After dissolving lipids in xylene, they were rehydrated through a series of graded ethanol solutions. They were stained with 1% cresyl violet, differentiated in 70% ethanol containing anhydrate acetic acids and then dehydrated with 95% and 100% ethanol. Samples were observed with a microscope (Model Axioskop, Zeiss, Jena, Germany). To estimate total pyramidal and granule cell numbers, photographs (12 per section) were randomly taken from the hippocampus at a magnification of ×400, and the stained cells were counted within an 80 × 80 μm dissector frame using image-analyzing software Win ROOF (ver. 5.6, Mitani Co., Fukui, Japan). To estimate the volume of the entire hippocampus, the Cavalieri principle was used (Reilly et al. 2003). The hippocampal structure was outlined and the computed areas were then summed and multiplied with the thickness and with the intersection distance.
5-Bromo-2′-deoxyuridine labeling
To measure the effect of chronic restraint stress on survival and differentiation of newly divided cells in the DG, 5-bromo-2′-deoxyuridine (BrdU) labeling was carried out 1 day before starting the chronic restraint stress. BrdU was purchased from Sigma-Aldrich (St Louis, MO, USA) and dissolved in saline. BrdU (75 mg/kg) was injected intraperitoneally (i.p.) three times at 2-h intervals as described previously (Ibi et al. 2008).
After stress, mice were deeply anesthetized with diethyl ether, and transcardially perfused with ice-cold saline followed by 4% paraformaldehyde in 0.1 M phosphate-buffered saline (PBS, pH 7.4). The brain was post-fixed in the same fixative and cryoprotected in 0.1 M PBS (30% sucrose). The brain was embedded in Tissue-Tek O.C.T. compound (Sakura Finetech, Tokyo, Japan) and stored at −80°C. Every fifth coronal section (30 μm) was collected between stereotaxic coordinates bregma −1.2 to 3.0 according to the brain atlas (Paxinos and Franklin 2004). For BrdU single-staining, sections were treated with 0.1% Nonidet-40/0.01 M PBS (pH 7.4) overnight (4°C) and denatured in a microwave oven in 0.01 M citrate buffer (pH 6.0). After blocking in 10% goat serum/PBS with 0.1% Nonidet-40 (Sigma-Aldrich), BrdU-positive cells in the sections were detected using BrdU labeling and detection kit 2 (Roche Diagnotics GmbH, Mannheim, Germany) according to the manufacturer's instructions. For double-staining of BrdU/neuronal nuclei (NeuN) and BrdU/glial fibrillary acidic protein (GFAP), sections were pre-treated with 1 M HCl, followed by 0.1 M borate buffer and then washed in PBS before blocking. Rat anti-BrdU antibody (1 : 200; Abcam, Cambridge, UK), mouse anti-NeuN antibody (1 : 100; Millipore, Billerica, MA, USA) and mouse anti-GFAP antibody (1 : 1500; Sigma-Aldrich) diluted in PBS (0.1% Triton X-100 and 5% goat serum) were applied to sections [overnight, 4°C and 6 h, room temperature (RT)]. After washing, goat anti-rat Alexa 568 and goat anti-mouse Alexa 488 (1 : 1000; Invitrogen, Eugene, OR, USA) antibodies were applied to sections (2 h, 25°C). For Ki67 or doublecortin (DCX) single-staining, sections were treated with 0.5% hydrogen peroxide in methanol to block endogenous peroxidase activity, and denatured in the microwave oven. They were incubated in blocking solution (10% goat serum and 0.1% Triton X-100 in 0.01 M PBS, 1 h), and then with rabbit anti-Ki67 (1 : 2000; Novocastra, Newcastle, UK) or rabbit anti-DCX (1 : 500; Abcam) antibody (overnight, 4°C). They were washed and incubated with biotinylated goat anti-rabbit antibody (1 : 200; Vector Laboratories, Burlingame, CA, USA, 1 h, 25°C). The sections were washed and processed with avidin-biotinylated horseradish peroxidase complex (Vector ABC kit, Vector Laboratories), and the reaction was visualized using diaminobenzidine. For double-staining of Npas4/several cell markers, coronal sections (10 μm) were incubated with blocking solution (3% bovine serum albumin, 10% donkey serum and 0.3% Triton X-100 in 0.01 M PBS, 1 h, 25°C), and then with primary antibodies (overnight, 4°C). They were washed with 0.01 M PBS and incubated with secondary antibodies (2 h, 25°C). Sections were mounted in fluorescent medium (S3023; Dako Cytomation, Kyoto, Japan). The following antibodies were used: rabbit anti-Npas4 (1 : 32), mouse anti-NeuN (1 : 200; Millipore), mouse anti-Calbindin D-28 (1 : 500; Swant, Bellinzona, Switzerland), goat anti-DCX (1 : 50; Santa Cruz Biotechnology, Santa Cruz, CA, USA), goat anti-Sox-2 (1 : 100; Santa Cruz Biotechnology), Alexa 488-conjugated donkey anti-rabbit (1 : 500, Invitrogen), Alexa 594-conjugated goat anti-mouse (1 : 500, Invitrogen) and Alexa 594-conjugated donkey anti-goat (1 : 500, Invitrogen). The rabbit anti-Npas4 antibody (Japan Bio Services Co., Ltd., Saitama, Japan) was raised against a carboxy-terminal region of Npas4 (amino acids 622–635). For pre-absorption experiment, Npas4 antibody was incubated with a 5-fold excess of the antigen peptide (overnight, 4°C).
Quantification of immunostaining cells
Every fifth section throughout the hippocampus (total 12 sections from each mouse) was processed for BrdU, Ki67 and DCX immunohistochemistry. All immunostained cells in DG were examined using a light microscope (Axio Imager; Zeiss) and counted by a blind experimenter. All counts were performed at 400× magnification (objective; 40×). To obtain the total number of cells per DG, we multiplied the counted number of positive cells by five. Double-stained cells were imaged and quantified using a confocal laser scanning microscope (LSM 510; Zeiss). Each cell was analyzed along the entire 'z' axis. Ratios of BrdU-positive cells co-labeled with NeuN or GFAP among BrdU-positive cells were determined.
Quantitative analyses of Npas4 mRNA by real-time RT-PCR
Mice were decapitated and brains were removed immediately after restraint stress or 4 h after corticosterone injection (10 mg/kg, s.c., in corn oil, Sigma-Aldrich). For DG, tissues from two animals were pooled as one sample and for hippocampus tissue from one animal used as one sample. The total RNA isolated from the DG or hippocampus (RNeasy Mini Kit, Qiagen, Hilden, Germany) was converted into cDNA using SuperScript III First-Strand Synthesis System for RT-PCR (Invitrogen) and quantified using a 7300 real-time PCR System (Applied Biosystems, Foster City, CA, USA). Quantitative real-time RT-PCR was performed in a volume of 25 μL with 0.5–1.0 μg of cDNA and 0.5 μM primers in the Power SYBR Green Master Mix (Applied Biosystems). The primers used were as follows: 5′-AGCATTCCAGGCTCATCTGAA-3′ (forward) and 5′-GGCGAAGTAAGTCTTGGTAGGATT-3′ (reverse) for Npas4, 5′-TGTCAAGCTCATTTCCTGGTATGA-3′ (forward) and 5′-CTTACTCCTTGGAGGCCATGTAG -3′ (reverse) for glyceraldehyde-3-phosphate dehydrogenase, 5′-CGATGCCCTGAGGCTCTTT-3′ (forward) and 5′-TGGATGCCACAGGATTCCA -3′ (reverse) for β-actin used as internal controls.
In situ hybridization
Npas4 mRNA level in tissue was detected with an in situ hybridization kit (Nippongene, Tokyo, Japan) following the manufacturer's manual with modifications. Briefly, immediately after restraint stress, mice were deeply anesthetized with diethyl ether, and transcardially perfused with ice-cold saline followed by 4% paraformaldehyde in 0.1 M PBS (pH 7.4). The brains were removed, post-fixed in the same fixative and then cryoprotected in 12%, 15% and 18% sucrose in Hanks' Balanced Salt Solution (HBSS) buffer (pH 7.4). The brains were embedded in Tissue-Tek O.C.T. compound (Sakura Finetech) and stored at −80°C. Thick coronal brain sections of 10 μm were cut and mounted. Nucleotides 1485–2409 of Npas4 (BC_129861) were used as a template for transcription and labeling of Npas4 antisense and sense with digoxigenin-UTP (Roche Diagnotics GmbH). Sections were hybridized with probes and mRNA signals were visualized by enzyme-catalyzed color reaction with an NBT/BCIP kit (Roche Diagnotics GmbH, 16 h, 25°C) in a dark room.
Statistical analysis
Statistical analyses were performed using SigmaStat 3.1 software (Systat Software, Inc., Chicago, IL, USA). For body weight gain, differences among groups were analyzed by repeated measures anova. Differences among groups were analyzed by Student's t-test or one-way anova, followed by Bonferroni's post hoc test.
Effect of chronic restraint stress on body weight gain and blood corticosterone level, hippocampal volume and the number of Nissl-stained cells in the hippocampus
To explore whether the mice that received 4-week chronic restraint stress revealed biologically significant features, we measured stress responses during and after the stress. During the period of 4 weeks of stress, chronic restraint stress significantly inhibited the body weight gain [0 day, control group, 19.5 ± 0.1 g; stress group, 19.7 ± 0.1 g; 24th day, control group, 40.1 ± 0.4 g; stress group, 31.9 ± 0.7 g, treatment, F (1, 21) = 165.719, p < 0.001; days, F (8, 168) = 717.763, p < 0.001; treatment and day interaction, F (8, 168) = 41.147, p < 0.001, Fig. 1a]. Blood corticosterone level after the final restraint stress was significantly increased in stressed mice compared with that of the control group (p < 0.05, Fig. 1b). To determine whether chronic restraint stress influences hippocampal volume and the number of hippocampal cells, we performed Nissl staining. Chronic restraint stress significantly decreased the hippocampal volume by 6.3% (p < 0.05), but there was no significant difference in the number of Nissl-stained cells in the hippocampus (Table 1, Figure S1).
|Hippocampal volume (mm3)
|Nissl-stained cells (×102 counts/mm2)
|6.98 ± 0.17
|42.01 ± 0.53
|40.64 ± 1.42
|119.96 ± 1.24
|6.54 ± 0.11*
|41.50 ± 1.03
|37.64 ± 1.19
|115.55 ± 2.1
Effect of chronic restraint stress on associative fear memory
Mice were subjected to the conditioned fear memory test to examine the function of hippocampus after chronic restraint stress for 4 weeks. Contextual fear memory requires both amygdala and hippocampus (Phillips and LeDoux 1992; Anagnostaras et al. 1999), whereas tone-dependent memory requires the amygdala but not the hippocampus. In the context-dependent test, mice were returned to the conditioning box, and then their freezing behavior was analyzed for 2 min. The mice subjected to chronic restraint stress exhibited significantly less freezing behavior than control mice (p < 0.05, Fig. 1c). On the other hand, there was no significant difference in freezing time between control and stressed mice when the freezing behavior was analyzed in the neutral cage in the presence of a continuous-tone stimulus identical to the conditioned stimulus (Fig. 1d). Mice were tested for foot shock threshold to be sure that any alterations in memory were not because of changes in sensitivity to foot shock. There was no significant difference in the pain threshold of the chronic stress mice (0.27 ± 0.02 mA, n = 16) compared with that of the control mice (0.29 ± 0.02 mA, n = 16).
Effect of chronic restraint stress on hippocampal neurogenesis
As recent studies have suggested that neurogenesis plays an important role in hippocampus-dependent memory (Leuner et al. 2006; Imayoshi et al. 2008; Kitamura et al. 2009) and chronic restraint stress exposure led to memory deficit in a hippocampus-dependent manner, we examined the effect of chronic restraint stress on cell proliferation in the DG of the hippocampus. Ki67-positive cells, for which the antigen is expressed in all active parts of the cell cycle, G1, S, G2 and M (mitosis) phases but not G0 phase (Braun et al. 1988), were mainly observed in the SGZ rather than other areas as either clusters or single cells (Fig. 2a). Chronic restraint stress significantly decreased the number of Ki67-positive cells in the SGZ by 17% (p < 0.05) and thereby reduced that in the DG by 16% (p < 0.05, Table 2).
|Control group
|Stress group
|2628.3 ± 77.0
|177.5 ± 19.0
|259.2 ± 49.8
|2191.7 ± 64.4
|2200.0 ± 150.6*
|129.2 ± 16.1
|252.5 ± 28.4
|1818.3 ± 136.1*
|1241.0 ± 107.9
|134.8 ± 12.3
|792.8 ± 74.5
|313.5 ± 33.7
|895.2 ± 49.7*
|118.1 ± 9.7
|533.9 ± 43.8**
|243.2 ± 26.2
|11 555.1 ± 479.2
|33.5 ± 11.9
|1271.7 ± 173.3
|10 249.9 ± 309.8
|9993.3 ± 327.0*
|36.7 ± 10.7
|1090.8 ± 170.1
|8865.8 ± 343.0*
5-Bromo-2′-deoxyuridine is incorporated into the DNA of proliferating cells during the S phase of the cell cycle (Rakic 2002) and has been utilized in a number of in vitro and in vivo studies to label neural progenitors. To measure the effect of chronic restraint stress on the survival of newly divided cells in the DG, BrdU was injected into mice 1 day before the chronic restraint stress was initiated, and the numbers of BrdU-positive cells were counted after 4 weeks of restraint stress. There was an apparent difference between control and stressed mice in the number and location of BrdU-positive cells in the DG of hippocampus (Fig. 2b). Chronic restraint stress significantly decreased the number of BrdU-positive cells in the GCL by 27% (p < 0.01) and thereby decreased that in the DG by 32% (p < 0.05, Table 2). DCX is one of the immature granule neuron markers and is useful to assess the rate of neurogenesis in the adult mammalian hippocampus (Liu et al. 2008). Chronic restraint stress significantly decreased the number of DCX-positive cells in the SGZ by 13% (p < 0.05) and thereby decreased that in the DG by 13% (p < 0.05, Fig. 2c, Table 2).
Finally, we investigated the effect of chronic restraint stress on the differentiation of newly divided cells. Mice were subjected to 4-week chronic restraint stress after BrdU injection, and then we measured NeuN-positive cells (neuron) and GFAP-positive cells (astrocyte) among BrdU-labeled cells in the DG of the hippocampus. There was no apparent difference in the distribution and the rate of NeuN- (Fig. 3a and b) or GFAP-positive cells (Fig. 3a and c) among BrdU-labeled cells in the DG of the hippocampus.
Effect of restraint stress on the expression levels of Npas4 mRNA in the dentate gyrus
To address whether restraint stress reduces Npas4 expressions in the DG of the hippocampus, mice were subjected to acute (2 h or 6 h) or chronic (6 h/day, 6 days a week for 4 weeks) restraint stress. The expression levels of Npas4 mRNA in the DG were analyzed by real-time RT-PCR. Both acute (2 h and 6 h) and chronic restraint stresses significantly decreased the levels of Npas4 mRNA compared with those in control mice [F (3, 16) = 8.718, p = 0.01, Fig. 4a]. In situ hybridization analysis revealed that intense signals that react with antisense Npas4 probes were detected in the pyramidal and granule cell layer in the hippocampus. There was no signal detected when sense probes were used, suggesting that Npas4 mRNA is abundant in the hippocampus (Fig. 5a). Chronic restraint stress markedly decreased the signals of Npas4 mRNA in the DG, CA3 and CA1 subregions of the hippocampus compared with those in control mice (Fig. 5b).
Effect of corticosterone treatment on the expression levels of Npas4 mRNA in the hippocampus
To address the relationship among stress exposure, reduction of Npas4 expression and the stress-induced impairment of neurogenesis and hippocampus-dependent fear memory, we examined the effect of corticosterone injection in mice on Npas4 mRNA levels in the hippocampus. The Npas4 mRNA levels in the hippocampus of mice following corticosterone injection were significantly reduced to 70.0% (p < 0.05) that of vehicle-treated control mice (Fig. 4b).
Phenotype of Npas4-positive cells in the dentate gyrus of the hippocampus
Finally, to characterize the Npas4-expressing cells in the DG, we performed double immunostaining for Npas4 and several cell markers such as SRY-related HMG box 2 (Sox-2), DCX, calbindin and NeuN. Antibody pre-absorption with the antigen peptide completely blocked the Npas4 staining in the DG (Figure S2). Sox-2 is a marker for both quiescent neural progenitors and amplifying neural progenitors (Segi-Nishida et al. 2008). DCX is a marker for immature neurons, while calbindin and NeuN are mature neural markers in the DG. At 7 weeks old, most Npas4 immunoreactivities were co-localized with the cells positive for NeuN in the GCL of control mice (Figs 6a and S2), while some of them partially co-localized with the cells positive for Sox-2 (Figs 6b and S2), DCX (Figs 6c and S2) in the SGZ and calbindin in the GCL (Figs 6d and S2). Similar results were observed in the stressed mice (Figs 6e–h and S2). Immunoblot studies revealed that the protein expression level of Npas4 in DG was not changed significantly in stressed mice (data not shown).
We demonstrated in the present study that chronic restraint stress after weaning decreased cell proliferation, survival of newly divided cells and neurogenesis in the DG. Numerous studies have reported that stress disrupts hippocampal neurogenesis. Cell proliferation in the DG is decreased by a variety of stress paradigms, including restraint stress (Nagata et al. 2009; Veena et al. 2009), subordination stress (Yap et al. 2006), social isolation (Dong et al. 2004), resident-intruder stress (Gould et al. 1998), water immersion restraint stress (Tamaki et al. 2008), chronic unpredictable stress (Heine et al. 2004), predator odor (Mirescu et al. 2004), sleep deprivation (Mirescu et al. 2006) and chronic mild stress (Alonso et al. 2004). Otherwise, the effect of stress on survival and differentiation of newly divided cells is more intricate than cell proliferation. Several studies have shown that the decrease of cell proliferation in the DG results in the suppression of neuronal generation (Pham et al. 2003; Westenbroek et al. 2004). Other reports have indicated that the influence of stress on hippocampal neurogenesis is very short-lived, and the decrease in the number of neuronal precursor cells is followed by an enhancement of cell survival, as the total number of new mature neurons appears unchanged (Tanapat et al. 2001; Malberg and Duman 2003). On the other hand, chronic stress after BrdU labeling decreases the survival of newly divided cells (Lee et al. 2006; Ibi et al. 2008). Our current findings are generally consistent with previous studies. Indeed, the stress exposure paradigm in this study was sufficient to increase serum corticosterone level in stressed mice compared with that in control mice, which may affect the fate of newly generated neurons in the DG. Meanwhile, chronic restraint stress had no apparent effect on the distribution and the rate of NeuN- or GFAP-positive cells among BrdU-labeled cells in the DG of the hippocampus. These results suggest that differentiation of newly divided cells in the hippocampus may be unaffected by chronic restraint stress after weaning in mice; however, we should also investigate the effect of chronic restraint stress on microglial cells.
We also found that chronic restraint stress significantly decreased the hippocampal volume without affecting the number of Nissl-stained cells. It has been reported that chronic restraint stress decreased the length and branch of apical dendrites in the CA3 subregion (Watanabe et al. 1992; Magariños and McEwen 1995). Therefore, atrophy of the apical dendrites of CA3 pyramidal neurons and the decrease of neurogenesis in the DG may be related to the reduction of hippocampal volume.
As regards the functional changes in the hippocampus, we demonstrated that chronic restraint stress impaired context-dependent fear memory in mice. Even though there is a report that chronic restraint stress enhanced fear memory (Conrad et al. 1999), it is generally accepted that stress impairs memory in rodents and the increased levels of circulating glucocorticoids inhibit memory in rodents (Lemaire et al. 2000; Montaron et al. 2006). The context-dependent fear memory is hippocampus-dependent (Phillips and LeDoux 1992) and the hippocampus is involved in acquisition and transient storage of context-dependent fear memory (Phillips and LeDoux 1992; Anagnostaras et al. 1999). Although a few studies have reported that contextual fear memory was not influenced by hippocampal neurogenesis (Shors et al. 2001; Zhang et al. 2008), irradiated rats or mice, ganciclovir-treated GFAP-tk mice and tamoxifen-treated Nes-CreERT2/NSE-DTA mice, all of which show defects of neurogenesis in the hippocampus, are defective in the conditioned fear memory test (Winocur et al. 2006; Imayoshi et al. 2008). In contrast to the context-dependent memory, tone-dependent fear memory was not significantly affected in chronic restrained mice. It is reported that tone-dependent fear memory is hippocampus-independent, but the lateral nucleus of the amygdala links the auditory conditioned stimulus with the fear response via the thalamo-amygdala or thalamo-cortico-amygdala pathway (LeDoux et al. 1990; Romanski and LeDoux 1992). Taken together, our results suggest that chronic restraint stress in mice, which leads to the hypersecretion of glucocorticoids, may disrupt the hippocampal function as evidenced by the impairment of context-dependent fear conditioned memory, which is associated with the inhibition of neurogenesis in the hippocampus and reduced hippocampal volume.
To clarify the putative molecular candidates for the stress-induced memory impairment and morphological changes in the hippocampus, we examined whether the expression level of Npas4 was regulated by stress and circulating corticosterone level. Acute or chronic restraint stress exposure as well as acute corticosterone injection significantly reduced the mRNA levels of Npas4 in the DG and the hippocampus, respectively, suggesting that stress may decrease Npas4 expression in the hippocampus through the action of circulating corticosterone level. It has been reported that Npas4 mRNAs are highly expressed in the hippocampus (Moser et al. 2004). Npas4 has constitutive or developmental functions that may be critical for regulating the transcriptional control of limbic patterning and function (Moser et al. 2004). Npas4 plays a role in the development of inhibitory synapses by regulating the expression of activity-dependent genes and appears to regulate a wide variety of genes modulating synaptic functions (Lin et al. 2008). Furthermore, we observed that Npas4 mRNA is present in neural stem/progenitor cells derived from the hippocampus of embryonic mice as originally described by Yoneyama et al. (2007, 2009) (data not shown).
Interestingly, BDNF is a target gene of Npas4 in mouse hippocampal primary cultures (Lin et al. 2008), and the expression level of BDNF is reported to be decreased by stress in the hippocampus (Vaidya et al. 1999; Xu et al. 2006). It is well known that BDNF positively regulates neurogenesis and plays a role in the differentiation and survival of neuronal progenitor cells (Sairanen et al. 2005; Scharfman et al. 2005). We assume that the inhibition of Npas4 gene expression in both progenitor cells and mature neurons may contribute to the decreased action of BDNF in the hippocampus. Taken together, chronic stress may reduce Npas4 expression in the DG of hippocampus through the action of corticosterone, leading to an impairment of hippocampal neurogenesis and hippocampus-dependent memory in mice. To further support the involvement of Npas4 in stress-induced impairment of neurogenesis, we showed that Npas4 was expressed in some of the Sox-2-positive cells, and most DCX- and calbindin-positive cells were colocalized with Npas4 in the hippocampus. These findings suggest that Npas4 is expressed from an early developmental stage of newly divided cells in the DG and that the down-regulation of Npas4 could be related to the impairment of neurogenesis by restraint stress.
Involvement of Npas4 in the hippocampal dysfunction through the impairment of neurogenesis induced by stress is also possible because neurogenesis modulates the hippocampus-dependent fear memory (Kitamura et al. 2009), in addition, newborn cells in adult DG may extend axons into the CA3 region, which release glutamates as their main neurotransmitter and form functional synapses with target cells (Toni et al. 2008). We revealed by in situ hybridization that stress decreased the mRNA expression level of Npas4 in DG, CA3 and CA1 subregions of hippocampus. Although we do not have any definitive evidence that the down-regulation of Npas4 mRNA expression level in CA3 and CA1 regions contributes to the reduced volume of hippocampus and impaired fear conditioned memory, Npas4 may play a role in synaptic and structural plasticity as well as hippocampal function in CA3 and CA1 regions because this transcription factor has been shown to regulate GABAergic synapse development in an activity-dependent manner (Lin et al. 2008) and to possibly play a role in dendritic-cytoskeleton modulation at synapses in the hippocampus (Ooe et al. 2004, 2009).
This study was supported in part by Grants-in-Aid for Scientific Research (Nos. 19390062 and 22390046) from the JSPS, the Global COE program from the Ministry of Education, Culture, Sports, Science and Technology of Japan, the Academic Frontier Project for Private Universities; matching fund subsidy from MEXT, 2007–2011, the Research on Risk of Chemical Substances, Health and Labour Science Research Grants supported by Ministry of Health, Labour and Welfare, Takeda Science Foundation, AstraZeneca Research Grant 2008, and JST, CREST.
Disclosure/conflicts of interest
The authors declare that there is no conflict of interest in the publication of this work.


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Home > News > 'Test tubes' far smaller than the width of a hair
August 30th, 2005
'Test tubes' far smaller than the width of a hair
Using a water droplet 1 trillion times smaller than a liter of club soda as a sort of nanoscale test tube, a University of Washington scientist is conducting chemical analysis and experimentation at unprecedented tiny scales.
The method captures a single cell, or even a small subcellular structure called an organelle, within a droplet. It then employs a powerful laser microscope to study the contents and examine chemical processes, and a laser beam is used to manipulate the cell or even just a few molecules, combining them with other molecules to form new substances.
University of Washington
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Good Riddance is finishing out their busiest year in ages with three California shows! San Francisco and Santa Cruz with Cobra Skulls and Western Addiction, and San Diego with Pulley and punk legends Battalion of Saints! Good Riddance's hometown paper, the Santa Cruz Weekly, gives the band their due, covering their beginnings (part 1), break up and reunion (part 2)
25Aug, 2016
Listen to a snippet of NOFX's audiobook with Tommy Chong narrating!
24Aug, 2016
More CJ Ramone tour dates added today!
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Chixdiggit continue to celebrate 25 years as a band! Read this interview with KJ and see them live!
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Watch as Sainte Catherines celebrate 10 years of Dancing For Decadence this year!
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PEARS & Direct Hit! tour starts today!
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toyGuitar are joining CJ Ramone this October!
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Chixdiggit! are celebrating 25 years as a band with their own beer!
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The Flatliners recorded an acoustic song for charity. Give it a listen and purchase!
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|January 4, 2013 10:30 AM EST
MOORESTOWN, NJ -- (Marketwire) -- 01/04/13 -- PayChoice announced its readiness to help its customers understand and comply with the new tax changes recently enacted by Congress, and signed into law by the President, known as the "American Taxpayer Relief Act of 2012" or ATRA. For well over two decades, PayChoice has always been at the forefront of all tax law changes.
The highlights of the ATRA law include the following:
- For 2013, Social Security tax rates for employees have returned to 6.2%, up from 4.2%. The tax applies to the first $113,700 of wages.
- The tax rates reduced in 2001 and 2003 for individuals earning up to $400,000 and families earning up to $450,000 will go unchanged per the Act. However, for Income above those levels, the tax rates will increase from 35% to 39.6%.
- The marriage penalty relief, which is the expanded 15% for those filing jointly, has been extended permanently.
Effective 2013 there is also a planned increase of 0.9% for employees known as the "Additional Medicare Tax." This is part of the Patient Protection and Affordable Care Act and is not part of ATRA. However this additional tax will impact individuals earning over $200,000 and married joint filers earning over $250,000. So regardless of filing status, the current Medicare tax rate of 1.45% will increase to 2.35% on earnings exceeding $200,000. The employer portion of 1.45% for Medicare will remain. Employers are now mandated to withhold the "Additional Medicare Tax" on all workers whose wages are over $200,000 regardless whether they are single or married.
Robert Digby, CEO of PayChoice, stated, "Our internal tax experts and customer support have always been exceedingly diligent in monitoring all pending federal, state, and local tax changes and the impacts these changes have on payroll, benefits, and human resources. We make every effort to maintain our systems as well as educate our customers and staff in the implications of these tax law changes."
About PayChoice
For more than 20 years, PayChoice has helped small- and medium-sized businesses (SMB) succeed by providing them a Fortune 500-style suite of payroll and HR services that frees the owner to focus on growing and managing their business.
The PayChoice suite of services includes payroll, tax filing, HR data management, time & attendance, workers' compensation and other employee management services and solutions. Because PayChoice delivers these services directly to SMBs through an intensely personal approach, our client satisfaction and retention are among the highest in the industry.
PayChoice is also the market leader in licensing payroll / HR software, and over 200 payroll / HR vendors nationally employ PayChoice's technology as the core of their service offering. All of PayChoice's applications are hosted in PayChoice's state-of-the-art private cloud infrastructure and delivered via a SaaS model. In total, PayChoice technology is used to meet the payroll and HR needs of over 150,000 businesses and nearly 2 million employees nationwide.
For more information, please visit www.paychoice.com or call 888.591.5151.
Chris J. Pfister
[email protected]
SYS-CON Events announced today that 910Telecom will exhibit at the 19th International Cloud Expo, which will take place on November 1–3, 2016, at the Santa Clara Convention Center in Santa Clara, CA. Housed in the classic Denver Gas & Electric Building, 910 15th St., 910Telecom is a carrier-neutral telecom hotel located in the heart of Denver. Adjacent to CenturyLink, AT&T, and Denver Main, 910Telecom offers connectivity to all major carriers, Internet service providers, Internet backbones and ...
Aug. 28, 2016 06:00 PM EDT Reads: 1,908
SYS-CON Events announced today that eCube Systems, a leading provider of middleware modernization, integration, and management solutions, will exhibit at @DevOpsSummit at 19th International Cloud Expo, which will take place on November 1–3, 2016, at the Santa Clara Convention Center in Santa Clara, CA. eCube Systems offers a family of middleware evolution products and services that maximize return on technology investment by leveraging existing technical equity to meet evolving business needs. ...
Aug. 28, 2016 05:30 PM EDT Reads: 698
DevOps at Cloud Expo – being held November 1-3, 2016, at the Santa Clara Convention Center in Santa Clara, CA – announces that its Call for Papers is open. Born out of proven success in agile development, cloud computing, and process automation, DevOps is a macro trend you cannot afford to miss. From showcase success stories from early adopters and web-scale businesses, DevOps is expanding to organizations of all sizes, including the world's largest enterprises – and delivering real results. Am...
Aug. 28, 2016 03:15 PM EDT Reads: 3,481
Pulzze Systems was happy to participate in such a premier event and thankful to be receiving the winning investment and global network support from G-Startup Worldwide. It is an exciting time for Pulzze to showcase the effectiveness of innovative technologies and enable them to make the world smarter and better. The reputable contest is held to identify promising startups around the globe that are assured to change the world through their innovative products and disruptive technologies. There w...
Aug. 28, 2016 03:00 PM EDT Reads: 728
Internet of @ThingsExpo, taking place November 1-3, 2016, at the Santa Clara Convention Center in Santa Clara, CA, is co-located with 19th Cloud Expo and will feature technical sessions from a rock star conference faculty and the leading industry players in the world. The Internet of Things (IoT) is the most profound change in personal and enterprise IT since the creation of the Worldwide Web more than 20 years ago. All major researchers estimate there will be tens of billions devices - comp...
Aug. 28, 2016 01:45 PM EDT Reads: 3,646
DevOps at Cloud Expo, taking place Nov 1-3, 2016, at the Santa Clara Convention Center in Santa Clara, CA, is co-located with 19th Cloud Expo and will feature technical sessions from a rock star conference faculty and the leading industry players in the world. The widespread success of cloud computing is driving the DevOps revolution in enterprise IT. Now as never before, development teams must communicate and collaborate in a dynamic, 24/7/365 environment. There is no time to wait for long dev...
Aug. 28, 2016 01:00 PM EDT Reads: 2,412
SYS-CON Events announced today that StarNet Communications will exhibit at the 19th International Cloud Expo, which will take place on November 1–3, 2016, at the Santa Clara Convention Center in Santa Clara, CA. StarNet Communications' FastX is the industry first cloud-based remote X Windows emulator. Using standard Web browsers (FireFox, Chrome, Safari, etc.) users from around the world gain highly secure access to applications and data hosted on Linux-based servers in a central data center. ...
Aug. 28, 2016 12:15 PM EDT Reads: 836
Traditional on-premises data centers have long been the domain of modern data platforms like Apache Hadoop, meaning companies who build their business on public cloud were challenged to run Big Data processing and analytics at scale. But recent advancements in Hadoop performance, security, and most importantly cloud-native integrations, are giving organizations the ability to truly gain value from all their data. In his session at 19th Cloud Expo, David Tishgart, Director of Product Marketing ...
Aug. 28, 2016 11:45 AM EDT Reads: 670
Data is the fuel that drives the machine learning algorithmic engines and ultimately provides the business value. In his session at Cloud Expo, Ed Featherston, a director and senior enterprise architect at Collaborative Consulting, will discuss the key considerations around quality, volume, timeliness, and pedigree that must be dealt with in order to properly fuel that engine.
Aug. 28, 2016 11:30 AM EDT Reads: 1,949
19th Cloud Expo, taking place November 1-3, 2016, at the Santa Clara Convention Center in Santa Clara, CA, will feature technical sessions from a rock star conference faculty and the leading industry players in the world. Cloud computing is now being embraced by a majority of enterprises of all sizes. Yesterday's debate about public vs. private has transformed into the reality of hybrid cloud: a recent survey shows that 74% of enterprises have a hybrid cloud strategy. Meanwhile, 94% of enterpri...
Aug. 28, 2016 11:00 AM EDT Reads: 3,118
There is growing need for data-driven applications and the need for digital platforms to build these apps. In his session at 19th Cloud Expo, Muddu Sudhakar, VP and GM of Security & IoT at Splunk, will cover different PaaS solutions and Big Data platforms that are available to build applications. In addition, AI and machine learning are creating new requirements that developers need in the building of next-gen apps. The next-generation digital platforms have some of the past platform needs a...
Aug. 28, 2016 11:00 AM EDT Reads: 684
Announcing @TelecomReseller Named "Media Sponsor" of @CloudExpo Silicon Valley | #IoT #Cloud #BigData
SYS-CON Events announced today Telecom Reseller has been named "Media Sponsor" of SYS-CON's 19th International Cloud Expo, which will take place on November 1–3, 2016, at the Santa Clara Convention Center in Santa Clara, CA. Telecom Reseller reports on Unified Communications, UCaaS, BPaaS for enterprise and SMBs. They report extensively on both customer premises based solutions such as IP-PBX as well as cloud based and hosted platforms.
Aug. 28, 2016 10:45 AM EDT Reads: 844
[session] Architecting for the Cloud By @RagsS | @CloudExpo @IBMBluemix #Cloud #Docker #Microservices
As the world moves toward more DevOps and Microservices, application deployment to the cloud ought to become a lot simpler. The Microservices architecture, which is the basis of many new age distributed systems such as OpenStack, NetFlix and so on, is at the heart of Cloud Foundry - a complete developer-oriented Platform as a Service (PaaS) that is IaaS agnostic and supports vCloud, OpenStack and AWS. Serverless computing is revolutionizing computing. In his session at 19th Cloud Expo, Raghav...
Aug. 28, 2016 10:30 AM EDT Reads: 883
The 19th International Cloud Expo has announced that its Call for Papers is open. Cloud Expo, to be held November 1-3, 2016, at the Santa Clara Convention Center in Santa Clara, CA, brings together Cloud Computing, Big Data, Internet of Things, DevOps, Digital Transformation, Microservices and WebRTC to one location. With cloud computing driving a higher percentage of enterprise IT budgets every year, it becomes increasingly important to plant your flag in this fast-expanding business opportuni...
Aug. 28, 2016 10:30 AM EDT Reads: 4,021
Enterprises have forever faced challenges surrounding the sharing of their intellectual property. Emerging cloud adoption has made it more compelling for enterprises to digitize their content, making them available over a wide variety of devices across the Internet. In his session at 19th Cloud Expo, Santosh Ahuja, Director of Architecture at Impiger Technologies, will introduce various mechanisms provided by cloud service providers today to manage and share digital content in a secure manner....
Aug. 28, 2016 09:30 AM EDT Reads: 766
StarNet Adds Secure Remote Linux and Unix Desktops to X-Win32 | @CloudExpo @XWin32 #Cloud #Linux #Security
StarNet Communications Corp has announced the addition of three Secure Remote Desktop modules to its flagship X-Win32 PC X server. The new modules enable X-Win32 to safely tunnel the remote desktops from Linux and Unix servers to the user's PC over encrypted SSH. Traditionally, users of PC X servers deploy the XDMCP protocol to display remote desktop environments such as the Gnome and KDE desktops on Linux servers and the CDE environment on Solaris Unix machines. XDMCP is used primarily on comp...
Aug. 28, 2016 09:30 AM EDT Reads: 721
Fact: storage performance problems have only gotten more complicated, as applications not only have become largely virtualized, but also have moved to cloud-based infrastructures. Storage performance in virtualized environments isn't just about IOPS anymore. Instead, you need to guarantee performance for individual VMs, helping applications maintain performance as the number of VMs continues to go up in real time. In his session at Cloud Expo, Dhiraj Sehgal, Product and Marketing at Tintri, wil...
Aug. 28, 2016 07:30 AM EDT Reads: 790
SYS-CON Events announced today that Isomorphic Software will exhibit at DevOps Summit at 19th International Cloud Expo, which will take place on November 1–3, 2016, at the Santa Clara Convention Center in Santa Clara, CA. Isomorphic Software provides the SmartClient HTML5/AJAX platform, the most advanced technology for building rich, cutting-edge enterprise web applications for desktop and mobile. SmartClient combines the productivity and performance of traditional desktop software with the simp...
Aug. 28, 2016 03:30 AM EDT Reads: 2,373
With so much going on in this space you could be forgiven for thinking you were always working with yesterday's technologies. So much change, so quickly. What do you do if you have to build a solution from the ground up that is expected to live in the field for at least 5-10 years? This is the challenge we faced when we looked to refresh our existing 10-year-old custom hardware stack to measure the fullness of trash cans and compactors.
Aug. 28, 2016 02:00 AM EDT Reads: 1,787
Extreme Computing is the ability to leverage highly performant infrastructure and software to accelerate Big Data, machine learning, HPC, and Enterprise applications. High IOPS Storage, low-latency networks, in-memory databases, GPUs and other parallel accelerators are being used to achieve faster results and help businesses make better decisions. In his session at 18th Cloud Expo, Michael O'Neill, Strategic Business Development at NVIDIA, focused on some of the unique ways extreme computing is...
Aug. 28, 2016 01:45 AM EDT Reads: 2,170


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Fox Lake Boat Canvas Repair
1. Rietesel's Boat Covers & Tops
I have come across your post on several review sites. I know in the past I have reached out to you. My offer still stands to take care of any issu…
2. Canvas Works
39532 N Il Route 59Lake Villa, IL 60046
From Business: Custom covers that fit your boat & wallet. Hours: MON 8:00am - 5:00pm TUE 8:00am - 5:00pm WED 8:00am - 5:00pm THURS 8:00am - 5:00pm FRI 8:00am - 5:00pm SAT 8:00am -…
3. United Canvas Inc
25434 W Il Route 173Antioch, IL 60002
From Business: Boat & Truck Covers, Vinyl Boat Lettering, Convertible Tops, Custom Fabrication, Vinyl Repair, Bimini Tops, Upholstery, Awnings & Roll Tops. 3YEARS WORKMANSHIP, 5 Y…
4. United Canvas & Awning
25434 W Il Route 173Antioch, IL 60002
From Business: At United Canvas & Awning in Antioch, IL, the difference is in our work. We provide beautiful, eye-catching awnings, sturdy boat covers and custom canvas work at un…
5. Tri City Canvas Upholstery Inc
1250 N Rand RdWauconda, IL 60084
6. Richmond Canvas
10819 N Main StRichmond, IL 60071
7. A B Kelly Co
212 W Main St # 5Cary, IL 60013
8. Five Star Boat Center
I live in Wisconsin and generally like to purchase big ticket items close to home for the simple reason that you take it back to where you buy it f…
9. Mineola Marine Inc
Great services. I really like this place. It is very clean and nice customer service and they put in 110% work ethic.
10. Roberts Performance Marine
Robert's Performance Marine is one ofthe best run business in FOX LAKE. ROBERT, THE OWNER. IS AN RXPERIENCED SAILOR, ABLE TO HANDLE ANY TYPE OF …
11. Fox Lake Harbor
the service that they offer is (GREAT), reasonable, & you are meet with a smile.
12. Ben Watts Marina
Overcharged and underserviced. These guys are really shady to say the least
13. The Boat Yard
This place sucks balls my friends and I have the worst flipping experiences here my wave runners were supposed to be done i was in no hurry I gave t…
14. West Marine
2 W Grand AveFox Lake, IL 60020
15. Rpm
8 S US Highway 12Fox Lake, IL 60020
16. Dick's Marine Service
32 N Pistakee Lake RdFox Lake, IL 60020
17. Gordys Lakefront Marine
36 S US Highway 12Fox Lake, IL 60020
18. Boating Warehouse
123 Christopher WayFox Lake, IL 60020
19. Fox Lake Canvas Shop
119 S US Highway 12Fox Lake, IL 60020
20. Bauske Boat Basin
152 N US Highway 12Fox Lake, IL 60020
21. Donovan's Power Boat Repair
112 Lakeview AveFox Lake, IL 60020
22. Kings Island Marine
500 S US Highway 12Fox Lake, IL 60020
23. Water & Wheels
500 S Il Route 59Fox Lake, IL 60020
24. The Boating Warehouse
1222 S US Highway 12Fox Lake, IL 60020
25. Munson Marine
Fox Lake, IL 60020
26. Custom Canvas
Roy has helped me numerous time with re-upholstering my ski boat interiors, and with canvas covers. His company can even do complete boat restoratio…
27. Bald Knob Marina
515 Bald Knob RdJohnsburg, IL 60051
From Business: Bald Knob MarinaWe exclusively sell Formula Boats by Thunderbird. Representing Formula is actually like carryings three lines. They are comprised of the Sun Sports,…
28. Walker's ServiceServices
25103 W Rollins RdIngleside, IL 60041
From Business: Walker's Service is a family owned and operated auto repair shop with over 38 years of experience, ASE Certified service technicians are expert at diagnosing proble…
29. Tradewinds Marine Center Inc
38788 N Broadway AveAntioch, IL 60002
30. Bottoms Up Boat Repair
1397 Horizon Drive Mchenry, IL 60051


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Myrtle Beach Travel Guide
Welcome to Myrtle Beach!This WorldWeb.com travel and tourism guide for Myrtle Beach, featuring hotels and other accommodations, tours, attractions, events, shopping and more, will help you plan a successful visit to Myrtle Beach, South Carolina, U.S.A.
For lovers of golf or deep-sea fishing, Myrtle Beach is the perfect destination. Centrally located on the temperate Grand Strand coastal area of South Carolina, Myrtle Beach is a great mix of history and outdoor fun. Golf courses are practically everywhere, and Pine Lakes Country Club has been housing golfers since 1927. Museums, such as Hopsewee Plantation, provide a peek into the history of Myrtle Beach and the Grand Strand. Or, take in a performance at the Carolina Opry.
Tourist Attractions
Hopsewee Plantation is the birthplace of Thomas Lynch Jr, one of the signers of the Declaration of Independance.
Hotels & Motels
This luxurious Sheraton is attached to the Myrtle Beach Convention Center and is located in the city's business district.
Current & Upcoming Events [ View All ]
Looking for something to do in Myrtle Beach? Check out these current and upcoming events. If you know of an event that is not listed in the travel guide, please feel free to add an event listing (it's free).
- Color Vibe 5k
Sep 24 » Myrtle Beach
- Color Vibe 5K -- Myrtle Beach
Sep 24 » Myrtle Beach


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Lakers 109, Pacers 94
INDIANAPOLIS -- The Lakers improved to 2-0 in the Andrew Bynum 2010-11 era with a dominant victory, the team's rotation falling into place beautifully.
Pau Gasol was splendid again with Bynum backing him up, scoring 28 points with eight rebounds, four assists and two blocks. Kobe Bryant deferred early on -- he didn't score one of the Lakers' first 27 points -- and then took over at points in the second half, finishing with 31 points and six assists.
Neither Gasol nor Bryant had a turnover.
Bynum had three points and five rebounds in 17 minutes off the bench. But everyone continued to look very happy to have him around again.
The Pacers had beaten the Lakers on Nov. 28 during their coldest spell of the season. But their execution was far better this time to reach 3-1 on this six-game Eastern trip.
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- New custody law for equal time for dads begins today; some question law's relevance (8/28/16)5
- Ex-Southeast student gets probation for placing homemade sex video on porn site without woman's knowledge (8/24/16)13
- Marble Hill fires entire sewer department (8/23/16)5
- Bootheel lawmaker seeks probe into crop damage by illegal herbicide spraying (8/24/16)1
- Former alt-rock frontwoman tells how she found Christianity (8/29/16)2
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- Schnucks bans solicitors, including organizations like Salvation Army (8/24/16)38
- Newsmakers 2016: Liz Glastetter (8/15/16)
- Court ruling, state suggest businesses may apply use, sales tax to deliveries (8/24/16)2
- Scott City School District introduces new preschool program (8/26/16)1
Mickey Mouse communicates with Franklin Elementary
When Franklin Elementary Teacher Elizabeth Speicher assigned a letter-writing exercise to her 4th-grade class she wasn't optimistic that her students would actually receive a reply from the famous Walt Disney character, Mickey Mouse. But they did! "I was really surprised," said Ms. Speicher, an enthusiastic teacher who is completing her first year of teaching at Cape Public Schools. The assignment was to write a friendly letter to their favorite Walt Disney character. "The children wrote some beautiful letters. It was a fun project and we learned a lot about letter writing," the teacher explained. In addition to a hand-written reply from Mickey, which Ms. Speicher read to her class, each child received an autographed greeting card. Pictured displaying their cards and letter from Mickey are Ms. Speicher and members of her class. Also pictured are three examples of letters the children wrote to their favorite Disney characters.


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|Name: _________________________
|Period: ___________________
This quiz consists of 5 multiple choice and 5 short answer questions through Chapter 19 and Afterword.
Multiple Choice Questions
1. How long did Marina stay in the Oak Cliff apartment with Lee?
(a) 5 weeks.
(b) 1 week.
(c) 15 weeks.
(d) 3 weeks.
2. Where did Oswald stay at the start of Chapter 7?
(a) A rental house.
(b) His aunt's.
(c) His brother's.
(d) His mother's
3. Who was Jim Garrison interested in during his investigation of Kennedy's death?
(a) Herbert Ferrie.
(b) David Farrey.
(c) David Ferrie.
(d) Tom Clank.
4. When did Texas announce it would hold a public court of inquiry into the assassination?
(a) December 29, 1963.
(b) November 25, 1963.
(c) November 29, 1964.
(d) November 2, 1963.
5. What does Posner say did not exist from the interrogation?
(a) Civilian witnesses.
(b) Transcripts or recordings.
(c) Police witnesses.
(d) Traces in the police archives.
Short Answer Questions
1. How many controversial issues did Posner mention in Chapter 14?
2. What did the Warren Commission conclude about the three shots fired at Kennedy?
3. What does Posner say numerous conviction-less investigations brought Jim Garrison?
4. What does the Warren Commission think the bullet mark on the curb was caused by?
5. How many Americans believed Oswald acted alone according to a Gallop Poll presented in Chapter 17?
This section contains 222 words
(approx. 1 page at 300 words per page)


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Posted 2/10/2013 at 7:52 PM by Karrie
This ornament was so adorable. The quality was amazing and the personalization was wonderful.Looked great on my tree.
Sending the same ornament to different places? If you order more than one of an item and want them shipped to different addresses, please order them individually. We will ask you for the individual shipping addresses later in the ordering process.
Bear Picture Frame Ornament (Blue) Christmas Ornament
Personalized Christmas Ornament or Personalized Christmas Table Top Decoration or Personalized Picture Frame
This little polar bear is lounging on the top of this picture frame ornament that can either be hung on the tree, or use the easel back that comes on it to sit on a shelf or table. This blue frame is a great way to share your son's Christmas photo with family members who will cherish the image as he grows. Use this for baby's first Christmas, 2nd Christmas, 3rd Christmas or any other age that you want to commemorate. Because this frame is blue and white, this doesn't only have to be for Christmas. Use this personalized miniature frame ornament as a baby gift for a baby shower or as a decoration in your son's room, sitting on the shelf with his adorable image staring back at you. An adorable way to display the sonogram or ultrasound of your baby boy.
Material: Resin
Read what our customers say about our Personalized Christmas Ornaments and their shopping experience!Hover over testimonials to stop scrolling
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12/10/2015 - "We really enjoyed your one-of-a-kind, personalized ornaments. Thank you! Linda N."


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morguenoun [ C ]uk /mɔːɡ/ us /mɔːrɡ/ mainly US UK usually mortuary
Thesaurus: synonyms and related words
(Definition of "morgue" from the Cambridge Advanced Learner's Dictionary & Thesaurus © Cambridge University Press)
(Definition of "morgue" from the Cambridge Academic Content Dictionary © Cambridge University Press)
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a place in a desert where there is water and therefore plants and trees and sometimes a village or town


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Rahm Emanuel: Blame the GOP for oil spill
I was adopted — when I was 41
Can librarians fix Wikipedia's gender gap?
How to fall in love with a new city
Stephen Colbert mocks Karl Rove's new Clinton conspiracies, but doesn't stop there
Donald Trump's campaign manager explains Trump's 6-figure Clinton Foundation donation
Donald Trump called Hillary Clinton 'a bigot.' Here are two essential reactions.
Is Nikki Haley the better candidate?


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First Official Pic of Bilbo in 'The Hobbit' Released
Director Peter Jackson is kind enough to reveal the first official look of Martin Freeman as Bilbo Baggins in "The Hobbit: An Unexpected Journey". Also released via Entertainment Weekly are an official photo of Ian McKellen as Gandalf and a sneak peek of work-in-progress centering on Jackson and Freeman.
''He fits the ears and he's got some very nice feet,'' Jackson told EW about Freeman's portrayal of Bilbo. ''I think he's got the biggest hobbit feet we've had so far. They're a little bit hard to walk in, but he's managed to figure out the perfect hobbit gait.''
Speaking of McKellen who is surrounded by actors different from "Lord of the Rings" series, Jackson said, ''He's in fantastic form. In a way, his role in 'The Hobbit' has more technical difficulties than 'Lord of the Rings' did, because he has scenes with 14 smaller characters - obviously the dwarves and the hobbit are shorter. ...I remember saying to him [laughs], 'Look, this isn't 'Waiting for Godot' or 'King Lear'. This is 'The Hobbit'. This is the real thing."
The first of the "Hobbit" movie would hit theaters in December 2012. Jackson is crafting the movie in a way that it would fit the schedule. ''There's no way you can pace yourself for shoots like these,'' the director said. ''When we were going through the schedule for 'The Hobbit,' I felt a terrible drop in my stomach when I saw that we'd be shooting for 254 days. We're only 12 days short of 'The Lord of the Rings,' even though we're only doing two movies. When I saw that, I had to sort of pick myself up off the floor and carry on."
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BJP disapproves of restrictions, questions 'hurried' cremation
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Holding that "a nation in shock" is entitled to display its sense of despair, BJP today disapproved of "Emergency-like" restrictions against anti- gangrape protests and renewed its demand for a special Parliament session to adopt strict laws on crime against women.
Expressing "profound condolences" to members of the victim's family and saluting her courage, the party also questioned the "hush hush" manner in which she was cremated.
BJP said that a proper debate in Parliament on the issue would "certainly be able to tap upon the serious sense of distress and dismay which is sweeping the country" after the "supreme sacrifice of that young brave daughter" of the country.
The main opposition party assured that it would cooperate with the the government in all suitable legislative measures to strengthen laws relating to crime against women, in strengthening the judicial architecture to ensure the speedy justice to those who are victims and taking actions against the perpetrators of these heinous crimes.
"The government needs to come quickly with a proper structured response and to reflect upon the serious concerns of the country, it is imperative that a special session of Parliament be called at the earliest which we have been demanding for some time," BJP spokesperson Ravishankar Prasad
told reporters here.
Criticising the government for "converting Delhi into a chavni (cantonment)", he said this "unfair" act of authorities reminds of Emergency era and added that "a nation in shock is certainly entitled to display its sense of despair, anger and pain."
"...why should Delhi be sealed like this. Why should Delhi be converted into a chavni reminding ourselves what is happened during Emergency. This is very very unfair.
"Metro trains have been closed, people cannot move. I think, a nation in shock is certainly entitled to display its sense of despair, anger and pain. Thats what democracy is about. We don't appreciate it at all," he said.
... contd.
Editors' Pick
- In Kashmir, so-called solutions are riddled with contradictions and divisions
- Why personal, social and political self-identification of Dalits must count more than legal nomenclature.
- The draft surrogacy bill violates the fundamental right of people to choose modes of parenthood
- Realpolitik drives Myanmar's outreach to India and China
- Epidemics in India are seldom followed by a long-term response
- Pakistan army has a battle to win: The corruption within
- Anger of Irom Sharmila's supporters should not be dismissed as selfishness or cynicism


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Daily Archives: December 23, 2010
NL Central Preview: Catchers
The first in my series of previews of the National League Central Division. The system, I will use to determine the best team in the division will be to rank the starters at each offensive position, including top-5 starters and the closer. The best will receive 6 points, and down to just 1 point given out to the worst starting player at that position in the division. At the end … Continue reading
Posted in Team News Tagged 2010-11 Offseason, Albert Pujols, Geovany Soto, Humberto Quintero, Joey Votto, Jonathan Lucroy, Prince Fielder, Ramon Hernandez, Ryan Doumit, Yadier Molina Comments Off on NL Central Preview: Catchers