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Jimmy Song's Programming Blockchain workshop taught me about a lot more than bitcoin.Read More »
Cryptocurrency Hedge Fund: We'll Succeed, Even if Bitcoin Ends Up Defunct
Dan Morehead, chief executive officer (CEO) and co-chief investment officer of struggling cryptocurrency hedge fund Pantera Capital, recently claimed that even if bitcoin fails like Pets.com did during the dot-com bubble, the hedge fund would still see investors make money. According to CNBC, Morehead's reasoning was that cryptocurrencies are revolutionary …Read More »
IBM Presents its Take on Blockchain for Business
Evanston IL—CCN correspondent Eric Eissler covered a lecture at Northwestern University by Marie Wieck, IBM General Manager, Blockchain. Wieck covered the industry, the history of blockchain and talked about IBM's Hyperledger project and how to tackle the challenges of digitization such as food-chain supply safety and digital identity. IBM enters …Read More »
Dubai Employee Steals $200,000 in Cryptocurrency Embezzlement Fraud
According to Dubai-based newspaper Khaleej Times, a cryptocurrency trading platform in the city caught its own employee stealing cryptocurrency for his personal use. Both the company as well as the convict were kept anonymous in the entire story. The man was an expert in managing trading systems, which was why …Read More »
Nauticus Exchange ICO Begins Today! 35 Percent Bonus Coins Available
This is a paid-for submitted press release. CCN does not endorse, nor is responsible for any material included below and isn't responsible for any damages or losses connected with any products or services mentioned in the press release. CCN urges readers to conduct their own research with due diligence into the …Read More »
Blockchain and AR is Revolutionizing the Gaming Industry
Merging the virtual and real world is a development that is becoming common in the gaming ecosystem. Most of these new games involve the integration of game visuals and audio content with a player's environment in real time. This phenomenon is described as Augmented Reality (AR). Real-life Participation Working with …Read More »
Market Crashes, Binance Pivots and OpenBazaar Developments: This week in Crypto
What happened in crypto this week? Price Watch Bitcoin is down about 8% this week to $7,800.This has been attributed to low volume, FOMO, and the ICO hearing. The news isn't all bad though, several experts are saying now might be a good time to buy. Ethereum is down about 16% this …Read More »
Florida State Employee Arrested for Mining Cryptocurrency on Agency Infrastructure
According to a statement released by the Florida Department of Law Enforcement (FDLE), an employee at the Department of Citrus was found using state-owned computing hardware to mine cryptocurrencies. The perpetrator, Matthew McDemott, served as the information technology manager for the department, which overlooks the functioning of the citrus industry …Read More »
Bitcoin Slips to $7,840 as Sell Volumes Intensify, Cryptocurrency Market Slumps
The valuation of the cryptocurrency market, which hovered near the $470 billion mark merely two weeks ago, has declined to $313 billion, as major cryptocurrencies including bitcoin fell in value. Bitcoin Slumps Over the past 24 hours, the price of bitcoin fell from $8,600 to $7,840, by nearly $800. While …Read More »
Traditional Liechtenstein Bank Launches Cryptocurrency Investment Platform
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Katarzyna Tonecka1, Zofia Pilch1, Kavita Ramji1, Bartlomiej Taciak2, Lukasz Kiraga2, Magdalena Krol2, and Tomasz P Rygiel1*
1Department of Immunology, Center of Biostructure Research, Medical University of Warsaw, Banacha 1a, 02-089 Warsaw, Poland
2Department of Physiological Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Nowoursynowska Warsaw, Poland
Received date: May 09, 2017; Accepted date: June 07, 2017; Published date: June 19, 2017
Citation: Tonecka K, Pilch Z, Ramji K, Taciak B, Kiraga L, et al. (2017) Immune Cells as Targets and Tools for Cancer Therapy . Immunotherapy (Los Angel) 3:143. doi: 10.4172/2471-9552.1000143
Copyright: © 2017 Tonecka K, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Visit for more related articles at Immunotherapy: Open Access
Immune cells are deeply intertwined in the development and progression of cancer. Their physiological functions can be divided into two main aspects on one hand they respond against the tumour and on the other they can counteract these responses or even promote tumour growth. For that reason immune cells are an obvious tool of therapeutic interventions in cancer treatment. Successful use of checkpoint inhibitors and cell-based therapies in the last few years has rapidly increased interest in manipulation of immune cell for cancer therapy. In this commentary we review the potential of targeting or using of macrophages, T cells, dendritic cells and NK cells in the cancer therapy.
Macrophages representing the major component of the infiltrate of solid tumours are called tumour-associated macrophages (TAMs). In contrast to inflammatory macrophages, TAMs are described as "alternatively" activated macrophages as they undergo stimulation by cytokines and chemokines that are present in the tumour microenvironment including IL-4, IL-10 and CCL2. TAMs show low tumoricidal activity and they promote tissue remodelling and angiogenesis. TAMs also promote tumour development, its local invasion and spread to distant sites [1], thus may constitute as an interesting target for anti-cancer therapy. Particularly blockade of the tumour infiltration by TAM can inhibit tumor growth or progression. For example, blocking of the colony stimulating factor 1 receptor (CSF-1R) in mice with aggressive mammary cancer, reduces pulmonary metastases regulated by macrophages (Figure 1) [2].
Figure 1: Monocytes attracted by releasing CSF-1 and CCL2 cancer cells migrate to tumour mass and differentiate to tumour associated macrophages (TAM). Blockade of CCL2 binding inhibits tumour infiltration by monocytes. Tumour specific antigens (Ag) are taken up by dendritic cells (DC) and presented to T cells that can be activated and differentiate into cytotoxic T cells (CTL). Which are able to recognize the same antigen on target cells and promote their killing. Antibody against programmed cell death protein 1 (PD1) improves effectiveness of T cell responses by preventing triggering of PD-1 receptor. CAR T cell and CAR NK cells can recognize tumour antigens on tumour cells and kill positive cells. Natural Killer cells (NK) activated by cancer cells (cellular stress, IL-10 and TGF-β) recognize and mainly attack cancer cells by cytoplasmic granule release and death receptor-induced apoptosis (Fas-FasL). Their activity can be improved by IL-18, IL-15 treatment.
Furthermore, CCL2-blocking antibody reduces number of metastasis and prolongs the survival of tumour-bearing mice [3]. Whereas, manipulation of Wnt signalling can change TAM activation state and thereby impact tumour progression [4]. Application of class IIa Histone Acetyl Deacetylase (HDAC) inhibitors, increased tumour infiltration by CD11b+ cells, eventually differentiating into macrophages. The treatment had positive effect on tumour-vasculature, whereas reduced cancer cell proliferation [5]. Deletion of REDD1 gene in hypoxic TAMs increases glucose uptake and changes the macrophage metabolism towards glycolysis. This, causes competition between TAMs and tumour endothelial cells, and results in stabilization of tumour vasculature and reduction of metastasis [6,7]. An alternative approach to this is stimulating macrophages to increase T cell immune response [8].
This was done using anti-CD47 antibody, which increased phagocytosis of cancer cells by macrophages and increased priming of CD8+ T cells, accompanied by decreased priming of CD4+ T cells. Overall, reprogramming methods including: antibody-mediated activation of IL-10, delivery of Toll-like receptor (TLR) agonists and targeting intracellular signalling molecules in combination with T cell-enhancing checkpoint inhibitors, allows changing the pro-tumoural immune infiltrate to an anti-tumoural state. These results indicate that modulated macrophages may represent as an interesting option for novel immune therapy.
T cells play a key role in cell-mediated immunity that is why for over fifty years they have been investigated to harness their potential in cancer therapy. Tumour-infiltrating lymphocytes can generate antitumour immune responses and are normally associated with a positive prognosis; however immunosuppression can hamper its effects. Adoptively transferred T cells have the potential to target and destroy cancer cells, while engineered T cells can overcome tumour immune evasion [9]. Nevertheless, transferred autologous T cells are inefficient in completely rejecting a tumour [10,11]. Expression of novel, genetically engineered receptors in T cells, improves cells persistence after adoptive transfer and enhance tumour specificity. Two main types of T cell modification exist. The first is the expression of cloned tumour antigen-specific T cell receptor (TCR) that is expressed next to the endogenous TCR. For that, α and β chains of TCR are identified, isolated and transduced into recipient T cells. In combination with identification of patient-specific mutations, this technology can be used to tailor anticancer responses according to patients' tumour genetic makeup. The second type is the expression of chimeric antigen receptors (CARs), with high specificity for antigen recognition in an MHC-independent fashion (Figure 1) [12]. Furthermore, antigens recognized by CAR T cells do not need to be peptides, but can also be a glycolipids or carbohydrates. Since the first generation of CAR T cells, an enormous improvement has been made. Second and third generation of CARs contain not only the main ζ domain of TCR, but also one or more costimulatory domains of CD28, ICOS, or 41BB which provide complete activation signals for T cells. Such CAR T cells are more effective as antigen recognition not only in stimulation of proliferation and survival, but also in preventing T cell anergy [13].
B cell malignancies are the most commonly targeted tumour types by engineered T cells. Firstly, they are relatively common and express several conserved cell surface markers (e.g. CD19, CD20, CD22). Furthermore, circulating B cell tumours provide an easy access for engineered T cells that are intravenously infused. CD19 is the most common B cell target for engineered T cell therapies. is expressed almost exclusively on benign and most malignant B cells [14] and therefore, possible "on-target off-tumour" activity of cytotoxic T cells are limited [15].
However a common complication of CAR T cell therapy is the cytokine release syndrome (CRS), but this can be controlled with an efficient treatment e.g. anti–interleukin-6 receptor antibody [15]. The main factors that influence efficacy of genetically modified T cell therapy are the loss of the targeted antigen expression or the inability to sustain their persistence and activation [16]. An additional challenge remaining is the efficiency of tumour targeting by CAR T cells in solid tumours, where poor tissue penetration is a major problem. This is illustrated by the observation that local administration of CAR T cells had higher accumulation at tumour sites compared with the systemic administration [17].
To be efficient most immune anticancer responses must overcome the immunosuppressive tumour microenvironment. Strategies that remove or deplete suppressor cells in combination with adoptive T cell therapy might enhance anti-tumour responses in cancer immunotherapies. Checkpoint inhibitors act as immunological brake-removers and tumour cells can utilise these checkpoints to escape destruction by the immune system. The addition of programmed cell death protein 1 (PD1) monoclonal antibody enhanced the anti-tumour effects of CAR T cells in preclinical models, suggesting that engineered T cells like other immune cells undergo immune suppression[18]Future therapies will most likely be accompanied by multiple immunomodulatory modalities, as checkpoint inhibin, to further enhance their efficacy. An alternative immunostimulatory approach is the combination of engineered T cells with inhibitors of immunosuppressive enzymes such as arginase or indoleamine 2,3- dioxygenase (IDO) or other checkpoint regulators such as VISTA or CD200R [19,20]. However safety of such combination treatments would need to be carefully verified, particularly in view of current safety challenges of the CAR T cells technology. Additional concern of the use of engineered T cells is the off-target reactivity and cytokine-release syndrome (CRS). Rapid tumour clearance has been associated with CRS that occasionally leads even to fatalities. Engineering of gene-edited human CAR T cells have been shown using techniques, including CRISPR–Cas9, molecular 'switches' enabling greater control over the performance of engineered T cells in vivo and may improve safety [21]. Cells may be engineered to express pro-death signals that can be induced with an exogenous element. An example of the "off switch" is the inducible human caspase 9, leading to deletion of CAR T cells in the animal model [22]. Other strategies that improve safety are: generation of T cells with a dual CAR that recognizes two targets or cells with expression of target molecules for monoclonal antibodies that could be used to eliminate CAR T cells when necessary [12]. Successful clinical trials with genetically modified T cells provide a proof of principle that shows the potential of the "living drug" therapies. However, the challenges remaining are broader effectiveness in solid tumours and better regulation to increase safety.
Dendritic cells (DCs) are at the center of the immune system owing to their ability to control both immune tolerance and to react against cancer. DCs are an essential component of vaccination because of their capacity to capture, process, and present antigens by MHC class I molecules to enable antitumour CD8+ T cell activation (Figure 1). DCbased immunotherapy can be used for vaccination against cancer through various ways in targeted peptide/protein and nucleic-acidbased vaccines captured by DCs in vivo [23], vaccines composed of DCs and antibodies conjugated with antigens [24,25] and vaccines composed of ex-vivo-generated DCs that are loaded with antigens. Sipuleucel-T (Provenge) is the first DC-based immunotherapy approved for the treatment of hormone-refractory prostate cancer [26]. The next step from ex vivo-engineered DCs is to target DCs in vivo in patients (in vivo DCs targeting) [27]. Several methods have been explored to target DCs in vivo. These include tumour-associated antigens (TAA)-bound antibodies targeting receptors on DCs [28], oncolytic viruses expressing GM-CSF, CD40L and other immune stimulatory molecules such as bacterial proteins (HP-NAP) [29]. An antibody against a molecule expressed on DC (DEC205) tagged with NY-ESO-1, co-administered with TLR (an agonist as adjuvants) is currently in a phase-I trial and it has been shown to be feasible and safe [30]. Another attractive strategy is to generate and use oncolytic viruses secreting GM-CSF. The foundation for this strategy is that viral oncolysis will release neoantigens from the cancer cells that can be captured by the tumour-residing DCs, which are then activated by virus-derived GM-CSF. An oncolytic Herpes Simplex virus secreting GM-CSF (T-Vec) was recently approved for the treatment of melanoma. Another tool to improve immunostimulatory potential of DCs is to target immunosuppressive molecule such as PD-L1, CTLA-4 or IL-10 to neutralize inhibitory signals. Several preclinical tumour models show that CTLA-4 blockade after DC vaccination may indeed enhance DC vaccine–induced T-cell responses [31-33]. In addition, it has been shown that DC-based immunotherapy in combination with anti–CTLA-4 antibodies seems to be more effective than the use of these agents alone [32,34]. Currently, there is no evident clinical data on the combination of DC vaccination with anti–PD-1 antibodies, but they are under investigation and the first results are expected in the near future. Similarly as in the case of engineered T cells, the safety of such combinatorial treatments will require a thorough investigation to reduce side effects.
Vaccine-induced protective immunity can be modulated using strategies that reduce or block other inhibitory molecules such as, CD200 to enhance the T cell activation. It has been demonstrated that lack of CD200R signaling inhibits outgrowth of endogenous tumours [35]. An interesting way to enhance the CD200-CD200R blockade is to combine the blocking agent with TLR7 stimulation. This goal is encouraged by studies showing that, a lack of CD200 increases TLR7- dependent immune response [36]. Several other TLR ligands are currently being tested in clinical trials, including CpG oligodeoxynucleotides (ODNs) (TLR9 ligand) and polyinosinic-polycytidylic acid (polyI:C) (TLR3 ligand) [37]. NK cells in tumour immune-surveillance and therapy.
Natural killer (NK) cells are lymphocytes of the innate immune system, known for their ability to recognize and kill malignant cells even in the absence of preimmunization or stimulation. Thus, making NK cells another promising target for cell-based immunotherapy for cancer treatment. Despite the fact that NK cells represent only 5-15% of circulating human lymphocytes, they exert immediate antitumour effect by releasing cytolytic enzymes, inducing FasL and TRAIL-mediated apoptosis and antibody-dependent cellular cytotoxicity (ADCC) (Figure 1). Many tumours have developed strategies to escape the immune surveillance by NK cells. This may be due to inhibition of NK cells by self- histocompatibility antigens (i.e. HLA-G) or suppressive signals (i.e. NKG2D ligands) derived from tumours and tumour-associated immune cells, such as myeloid-derived suppressor cells (MDSC), Tregs and TAMs [38-41]. The impaired expression of natural cytotoxicity receptors (NCRs) and defective function of NK cells was observed in many haematological malignancies and solid tumours [38,42]. To increase the cytotoxic potential of NK cell-based therapies, new approaches have evolved towards the use of expanded or ex vivo stimulated allogeneic NK cells, NK cell lines and genetically engineered NK cells with increased expression of cytokines, antibody binding receptors and activating receptors. The most extensively studied cytokine utilized to expand and activate NK cells is IL-2. Antitumour potential of IL-2 stimulated NK cells was documented in clinical trials in patients with advanced metastatic cancers [43]. However, use of IL-2 as an activating agent may also trigger Tregs proliferation via stimulation of the receptors for IL-2 and IL-2Rα (CD25). Selective depletion of CD25+ cells by lymphodepleting agents and IL-2 fused with diphtheria toxin (IL-2DT) followed by haploidentical NK cells infusion increased complete remission rate in AML patients in comparison to standard IL-2 administration [44]. IL-15 is another cytokine essential for NK cell maturation and survival, which has demonstrated a marked synergy with IL-2 on the viability and proliferation of NK cells [45]. Unlike IL-2, IL-15 does not induce Treg cell proliferation and capillary leak syndrome, thus IL-15 may be safely administered with NK cells [40]. Preactivation of NK cells with a cocktail of cytokines such as IL-12, IL-15, and IL-18 demonstrated an increases in antitumour effects [46]. Initial clinical trials suggest that NK-cell based therapy is safe and feasible; nevertheless, there is still a need to optimize the manufacturing process of clinical grade NK-cell products. Early investigations using cell lines derived from NK cells, such as the NK-92 cell line has revealed cytotoxic activity towards a wide range of malignant cells and has been used as a source of NK cells in clinical trials [47-49]. The advantage of NK-92 cell line is the lack of inhibitory receptors such as KIRs and NKp44 [50]. On the other hand, NK-92 cells do not express FcγRIIIa receptor (CD16), thus are enable to mediate ADCC, an important mechanism of action for the use of anti-cancer antibodies in therapy. This defect can be reverted by the introduction of mRNA coding for the antibody-binding receptor CD16. CD16-expressing variants could be combined with antitumour monoclonal antibodies in the clinic [51,52].
Finally, the design of CAR-modified NK cells have enabled high affinity specific recognition of tumour antigens and enhanced the cytotoxic potential of NK cells and NK cell lines in antitumour therapies, especially targeting B-cell malignancies (anti-CD19, anti- CD20 CARs) [53,54]. CAR-based immunotherapy have been used also in solid tumours, such as neuroblastoma (anti-GD2 CAR) and breast cancer (anti-EpCAM CAR) [55,56]. Her-2, which is overexpressed in many types of human malignancies also represents as a promising target for CAR-expressing NK cell-based therapies. Further studies clarifying the antitumour potential of modified NK cells have to be initiated, in order to maximize their cytotoxic potential in NK cellbased therapies [55-59].
The combination of standard or novel anti-cancer therapy together with targeting of immune cells present in the tumour microenvironment constitutes the future of cancer treatment. Our current understanding of tumour development and interactions occurring between cancer cells and immune cells enables for more efficient immune manipulations. A parallel change of immunosuppressive tumour microenvironment with stimulation of effector cell-based responses will be the best combination of immunotherapy [60-64].

subset: commoncrawl


text: We're still working on a memorial garden in the backyard at my Charlie Girl's gravesite. Charlie was the world's best dog that we lost last summer to cancer. We're kinda waiting for spring to do the plantings. It will be mainly wildflowers, a memorial stone and a forsythia bush that DH just can't kill, no matter how hard he tries. I want to plant a smallish tree in the center of her garden. It can't have an extremely long and evasive root system because the septic tank is also in the backyard. It will have to compete somewhat for sunlight with the old hickory trees to the east of it. This can be a blessing for a young tree during the brutal summers. I really want a flowering japanese cherry, but not sure they grow here. My other choice would be a dogwood. Any other trees I should consider?


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'No Game No Life Zero' movie screening for two days in U.S.
Movie theaters in the U.S. gets ready for No Game No Life Zero anime movie that will run for two days.
Sentai Filmworks announced its plan with Azoland Pictures and Fathom Events to present the anime film "No Game No Life Zero" for two days in the U.S. in October. The press release also unveiled the motion picture's release will feature the original anime with its Japanese audio and English subtitles.
"No Game No Life Zero" will be first seen in movie houses on October 5. Meanwhile, Sentai Filmworks' English dubbed version will premiere on October 8. The attendees will also see special video content featuring the film cast and the crew of "No Game No Life Zero."
What awaits Sora and Shiro in the new film?
Crunchyroll described the film as a story that had occurred thousand years before Sora and Shiro were born.
A war which broke out dismantled the earth and threatened the lives of the human race. A young man named Riku deems that the remaining survivors can get through the war-led humanity.
Riku was in the ruins of an Elf city when he happened to meet Shivu, the female android who wanted Riku to tell her about the significance of having a human heart.
On the other hand, the original Yu Kamiya novel series centered on the life of the brother and sister, Sora and Shiro, who masked their identities as "Blank," an unbeaten duo of game geniuses. The game god challenged the siblings and lost. As a result, the god brought Shiro and Sora to Disboard, the real world that revolved around games. The siblings decided to conquer the sixteen ruling species and remain undefeated.
Cast, staff and important dates
Aside from the first release of "No Game No Life Zero" in film houses, the anime distribution companies will also introduce the movie at an event in Los Angeles known as the Anime Film Festival.
The motion picture adapted from the original "No Game No Life" series will premiere in North America in on September 17.
The anime with Japanese voice voice-and English subtitles will get featured on September 16 while its English dubbed recordings will hit the movie theaters on September 17.
Kyle Colby Jones, the director of numerous TV anime series and the TV anime version of "No Game No Life," will be the director for the film. Meanwhile, the voice actors include Scott Gibbs, Shannon Emerick, Kira Vincent-Davis and Caitlynn French.
More details regarding the upcoming event will be announced soon. The announcement will include the list of theaters where the movie will get shown as well as other information.


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Pet Sitter in Folsom
My name is Anissa and I am currently an Environmental Studies major in senior standing at California State University, San Bernardino. I am currently employed at the Student Recreational Wellness Center at my school working as a desk assistant for the Intramural Sports department. I am also an Exotic Animal Keeper at Forever Wild Exotic Animal Sanctuary located in Phelan Ca. When I'm not in class or work during the week, I spend my time working at the sanctuary caring for a large variety of exotic, barn and sometimes domestic animals. I also manage to volunteer once a week at the San Bernardino Humane Society, working in the recovery room. These commitments have taught me so much, from how to restrain specific animals in the appropriate and comfortable ways, to the knowledge of healthy and nutritious diets. Not only do I have a love for what I do, but I have a love for all animals and wish to pursue a career in wildlife rescue, rehabilitation and conservation. In other words, helping care for your animals would be a privilege!
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The Champ 1979
0 wallpapers
The more you love, the harder you fight.The world looks at Billy Flynn and sees a has-been who seemingly never was, an ex-boxing champion slammed to the mat years ago by booze and gambling. But Billy's son TJ sees what the world doesn't. He knows his flawed but loving father is, was and always will be The Champ.
Links: Posters, IMDb
1280 * 898 Lobby card with Jon Voight & Ricky Schroder
1280 * 1825 Lobby card with Jon Voight
1000 * 705 Publicity still of Jon Voight & Ricky Schroder


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- Use sunscreen with a sun protection factor, if you sweat a lot of suns, apply sunscreen after two hours, to protect your beauty.
- Make a wise selection of sunscreen, which gives you a wider protection. These types of sunscreens protect you from both types of ultraviolet rays, A and B. Always go for the option of nonacnegenic sunscreens, which keeps your pores clear.
- Apply more quantity of sunscreens, when you are exposed to more sun rays, like in midday, near water or beach and in hot summer.
- Sun rays can also affect your hairs and eyes, so use a hat and sunglasses.
- Some medications enhance the skin, sensitivity to the sun rays, like medicines for acne, if you are using such type of medicines, increases your skin protection.
- Sometimes you ignore to protect your ears, eyes and V on the chest, apply sunscreen to your these body parts as well, to gain full protection.
- Do not ignore your nose and feet, while protecting your skin from the sun.
How to protect your skin without sunscreen: Sunscreen the body through cancer-causing Ultra violet rays, therefore you need to slather this upon if you proceed outdoors. That is particularly true in the seaside and swimming pool -- drinking water and fine sand reveal sunshine, therefore you are prone to obtain burnt without having a safety. Should you remain the sunscreen in your own home, there is you don't need to lose out on any kind of outside Steps. You are able to avoid obtaining a sunburn through remaining in the tone and putting on protecting clothes.
Step 01
Put on long-sleeved clothing when you are outside. Trousers along with a long-sleeved clothing protect the body through sunshine; more dark colours safeguard pores and skin much better than gentle types. If you are in the seaside or swimming pool and cannot put on long-sleeved clothing, including your own body having a lengthy T-shirt.
Step 02
Clean clothing having a UV-repellent washing soap. These types of liquids layer clothes along with tiny deposits. The deposits avoid Ultra violet rays through infiltrating the material and dealing with the skin.
Step 03
Placed on the wide-brimmed loath prior to venturing out within the sunlight. That person uses up very easily, therefore you need to tone this having a loath, particularly if you aren't using sunscreen. Avoid using hay caps; they've little openings which allow sunshine via.
Step 04
Safeguard your own eye along with shades which filtration system UVA and UVB sun rays. Choose wrap-around shades for top safety.
Step 05
Sit down within the tone as frequently as possible when you are outside. Search for tone below a good awning or sapling. In the swimming pool or seaside, sit down below a sizable outdoor umbrella.
Step 06
Part heading outdoors between your several hours of 10 the. michael. in order to 3 g. michael. Sunshine tends to be most powerful during this period time period.
You will get the sunburn if it is overcast or chilly outdoors. You can also obtain burnt when you are underneath the drinking water. Ultraviolet rays may permeate cup.


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When people ask how I am I reply "okay" but the truth is I have a new okay, a sub-standard one compared to my old okay. Why? My second pregnancy was complicated by preterm Premature Rupture of the Membranes (pPROM) at just over 21 weeks resulting in the death of my son at just over 31 weeks. Since then I have had two miscarriages. This is the continuing story of my new okay.
Happy Birthday! Hope its full of delicious cake and one or two rocking presents!!


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WASHINGTON — Minnesota Gov. Jesse Ventura on Monday named an independent to replace the late Democratic Sen. Paul D. Wellstone, casting in doubt which party will control the Senate during the forthcoming lame-duck session of Congress.
Ventura's abrupt election-eve appointment of Dean Barkley, a fellow member of the Independence Party, means that the Senate could be divided among 49 Republicans, 49 Democrats and two independents when it meets early next week to consider important year-end legislation, including appropriations for the current fiscal year, creation of a Department of Homeland Security and executive-branch and judicial nominations.


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Saturday, July 11, 2009
Browser War
Browsers are used to access the internet and to retrieve the information on World Wide Web. NCSA Mosaic was the first web browser found in the year 1993 and then the Netscape Navigator browser was introduced to the market. At that time Microsoft launched internet explorer which is the number one browser till now. To compete with Internet explorer many browsers were introduced like Mozilla Firefox, Opera and Apple's Safari. Google also joined in the war of browsers. Recently it released the Google Chrome with lots of facilities. Security and speed are the very useful features of Google chrome. They have provided user friendly interfaces, which helps people to easily browse through. Let us wait and see who is going to win this browser war.
Posted by Basha at 10:45 AM
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Germ cell development and gametogenesis require genome-wide transitions in epigenetic modifications and chromatin structure. These changes include covalent modifications to the DNA and histones as well as remodeling activities. Here, we explore the role of the mammalian SWI/SNF chromatin-remodeling complex during spermatogenesis using a conditional allele of the ATPase subunit, brahma-related gene 1 (Brg1, or Smarca4). Not only do BRG1 levels peak during the early stages of meiosis, genetic ablation of Brg1 in murine embryonic gonocytes results in arrest during prophase of meiosis I. Coincident with the timing of meiotic arrest, mutant spermatocytes accumulate unrepaired DNA and fail to complete synapsis. Furthermore, mutant spermatocytes show global alterations to histone modifications and chromatin structure indicative of a more heterochromatic genome. Together, these data demonstrate a requirement for BRG1 activity in spermatogenesis, and suggest a role for the mammalian SWI/SNF complex in programmed recombination and repair events that take place during meiosis.
Mammalian gametogenesis requires numerous epigenetic changes to accompany the transition from somatic, diploid precursors to mature, haploid gametes (reviewed by Sasaki and Matsui, 2008). Embryonic epigenetic states are first erased and reprogrammed during the development of primordial germ cells (PGCs). Later, as germ cells differentiate and proceed through meiosis, their genomes undergo large-scale changes to histone and DNA modifications as well as chromatin structure. This transition is important for the progression through meiosis, which itself requires the action of macromolecular complexes to manage the series of events entailing meiotic recombination between homologous chromosomes. It is therefore not surprising that faithful execution of the meiotic program requires the action of a large number of epigenetic pathways. Recent work has identified DNA methyltransferases, histone-modifying enzymes, and RNA-binding proteins, which when inactivated by mutations, result in failure of spermatogenesis (Kota and Feil, 2010).
In both male and female germ cells, homologous recombination occurs during the first meiotic prophase. During this time DNA double-stranded breaks (DSBs) are induced, and repair at the DSBs generates DNA recombination between homologous chromosomes. Many of the factors required for repair of stress-induced DNA damage in somatic cells function during meiosis. In addition to their well-characterized roles in transcriptional regulation, chromatin-remodeling complexes have roles in DNA repair (Bao and Shen, 2007; Clapier and Cairns, 2009). Among them, members of the SWI/SNF ATP-dependent chromatin remodelers appear to have a conserved function in the DSB repair process in yeast (Chai et al., 2005) and human somatic cells (Park et al., 2006; Zhao et al., 2009). Upon induction of DNA damage by ionizing radiation, the SWI/SNF catalytic subunit BRG1 (SMARCA4 – Mouse Genome Informatics) is required for the phosphorylation of the histone variant γH2AX (H2AFX – Mouse Genome Informatics), which in turn promotes the recruitment of downstream repair proteins. These observations suggest a role for mammalian SWI/SNF remodeling complexes in the repair of stress-induced DNA damage; however, whether they also play a role in programmed DNA repair events, such as those during meiosis, is not yet known.
In this report, we show that a germline-specific ablation of Brg1 results in a critical defect in spermatogenesis in the mouse. We demonstrate that BRG1 protein levels normally peak during the early phases of meiosis I. Consistent with this pattern, germ cells lacking BRG1 fail to complete prophase I. Although γH2AX appears normally induced in mutant spermatocytes, it persists into later stages of prophase I along with aberrant foci of repair proteins. BRG1-depleted spermatocytes showed significant deviation in the expected global patterns of epigenetic modifications, including those that are associated with DNA damage and heterochromatin. Together, these results demonstrate an essential role for BRG1 in spermatogenesis, and suggest a function for BRG1-containing complexes in homologous recombination during meiosis.
Brg1 conditional deletion and genotyping
Mice carrying a floxed allele of Brg1 (Sumi-Ichinose et al., 1997) and Mvh-Cre (Gallardo et al., 2007) were maintained on a mixed background. Mutant and normal littermates were obtained from crosses between female homozygous Brg1flox mice with male Mvh-Cre/+; Brg1Δ/+ mice. PCR genotyping primers for wild-type and floxed alleles of Brg1 included: forward 5′-GCCTTGTCTCAAACTGATAAG and reverse 5′-GTCATACTTATGTCATAGCC; for the excised allele of Brg1, forward 5′-GATCAGCTCATGCCCTAAGG and the same reverse sequence; for Cre, forward 5′-CACGTGCAGCCGTTTAAGCCGCGT and reverse 5′-TTCCCATTCTAAACAACACCCTGAA. All animal work was carried out in accordance with IACUC protocols at UNC.
Histological analysis
After dissection, testes were fixed in Bouin's fixative overnight at 4°C, dehydrated in an ethanol series, and embedded in paraffin wax. Sections were made at 3 μm thickness using a microtome. Following standard protocols, sections were deparaffinized, rehydrated, and then stained with Hematoxylin and Eosin for histology.
Immunofluorescence staining and antibodies
Preparation of testis cryosections and nuclear surface spreads of meiocytes were performed as described previously (Peters et al., 1997; Kim et al., 2007) with the following modifications. For the preparation of cryosections, tissues were washed in PBS at pH 7.4, permeabilized in PBST (PBS with 0.1% Triton X-100), and fixed in 4% paraformaldehyde in PBST overnight at 4°C. Fixed tissues were saturated through a sucrose series and embedded in optimal cutting temperature (OCT; Sakura Rinetek, Torrence, CA, USA). Frozen tissue sections were cut at 8 μm thickness. Slides were dried for 2 hours on a 37°C warming plate or overnight at room temperature (RT). An antigen retrieval step was required for several antibodies: slides were washed in PBT (PBS with 0.1% Tween 20), boiled in 10 mM citric acid (pH 6.0) for 10–30 minutes, and cooled slowly at RT for 1 hour.
For the preparation of nuclear spreads, seminiferous tubules were dissected and incubated for 20 minutes in a hypotonic extraction buffer (30 mM Tris pH 8.2; 50 mM sucrose; 17 mM citrate; 5 mM EDTA, 0.5 mM DTT; 0.1 mM PMSF). The tubule fragments were added to 100 mM sucrose solution to release hypotonized nuclei by pipetting repeatedly or mincing with a blade. A drop of nuclear suspension was spread onto slides coated with 1% paraformaldehyde. Slides were dried completely, washed in 0.4% Photo-Flo solution (Kodak), and dried again for storage. We categorized substages of prophase I by localization of the synaptonemal complex proteins SYCP3 and SYCP1, together with the kinetics of histone variant γH2AX distribution (Mahadevaiah et al., 2001).
For immunofluorescence staining, slides were blocked in 5% serum from the same species used for the secondary antibody for 1 hour at RT, incubated in primary antibodies at an optimized titer overnight at 4°C and subsequently in Alexa fluorophore conjugated secondary antibodies (Molecular Probes) for 1 hour at RT. The M.O.M. Immunodetection Kit (BMK-2202, Vector) was used as directed to eliminate background staining from endogenous mouse immunoglobulin. Nuclei were counterstained with 4,6-diamidino-2-phenylindole (DAPI, Sigma). For imaging, slides were mounted using an anti-quenching medium, ProlongGold (Invitrogen, P36930). Fluorescence microscopy was conducted using Leica DMLB or Zeiss Imager-M2 microscopes. Antibodies used in this study are: rabbit anti-BRG1 (1:100, Santa Cruz, sc-10768), monoclonal mouse anti-BRG1 (1:1000, Santa Cruz, sc-17796), rabbit anti-BRM (1:250, Abcam ab15597), rabbit anti-beta-galactosidase (1:500, Cappel #55976), rabbit anti-CRE (1:200, Covance, MMS-106R), mouse anti-γH2AX (1:800, Millipore #05-636), rabbit anti-phospho-HH3 (1:500, Cell Signaling #9701), mouse anti-histone H3 dimethylated lysine 9 (1:500, H3K9me2, Abcam ab1220), rabbit anti-histone H3 acetylated lysine 9 (1:500, H3K9Ac, Millipore #06-942), rabbit anti-histone H4 pan-acetylated (1:200, Millipore #06-866), rat anti-HP1β (1:500, Abcam ab10811), rabbit anti-HP1γ (1:500, Abcam ab66617), mouse anti-MLH1 (1:500 BD BioScience #51-1327GR), rabbit anti-MVH (1:5000, Abcam #13840), mouse anti-PLZF (1:100, Calbiochem #OP128), rabbit anti-RAD51 (1:250, Calbiochem PC130T; 1:100, Santa Cruz sc-8349), rabbit anti-RPA (1:100, Bethy Lab IHC-00409), rabbit anti-SCP1 (1:1000, Abcam ab15090), rabbit anti-SCP3 (1:500, Abcam ab15093), mouse anti-TRA98 (1:500, Cosmo Bio 73-003-EX). Secondary antibodies used were goat or donkey IgG conjugated with one of Alexa Fluor 488, 568 and 594 (Invitrogen). When applicable, we used secondary antibody matched for the isotypes.
Cell proliferation and apoptosis assay
Proliferating cells were detected by immunostaining with a mitotic marker. Histone H3 phosphorylated at serine 10 (pHH3) specifically labels cells in metaphase. For cell apoptosis analysis, fluorescent TUNEL assay was conducted by using the In Situ Cell Death Detection Kit (Roche, #11 684 795 001).
Statistical analyses
Chi-squared analysis was carried out to test differences in meiotic stage distribution in mutant and wild-type spermatocytes. A two-sided Fisher's exact test was used to determine significance of SYCP3, RAD51, RPA (RPA1 – Mouse Genome Informatics), MLH1 distribution patterns. At least two mice per genotype were used for each experiment.
BRG1 expression in male germ cells
To determine whether BRG1 plays a role in gametogenesis, we analyzed BRG1 expression during spermatogenesis by immunofluorescence staining. BRG1 was detected as early as 3 days post partum (dpp), when the prospermatogonia, the quiescent precursors to the spermatogonial stem cells, re-enter the cell cycle (Fig. 1A). BRG1 expression continued at 7 dpp (Fig. 1B) as prospermatogonia differentiate into spermatogonia marked by the spermatogonial stem cell marker PLZF (ZBTB16 – Mouse Genome Informatics) (Buaas et al., 2004; Costoya et al., 2004) (inset, Fig. 1B). BRG1 expression remained at 12 dpp as spermatocytes proceeded with meiosis (Fig. 1C). However, in adult testes, where all stages of spermatogenesis are present, BRG1 was detected predominantly in SYCP3-expressing meiocytes (Fig. 1D). BRG1 levels increased from zygonema to pachynema (Fig. 1E), during which DNA repair and recombination take place. We further analyzed the expression of the BRM ATPase subunit expression during spermatogenesis. In contrast to BRG1, it is expressed in somatic cells and differentiating spermatids, but not in wild-type spermatocytes (supplementary material Fig. S1). Thus, BRG1 and BRM may have complimentary, non-redundant roles during male meiosis. Together, these observations demonstrate that BRG1 is dynamically regulated during spermatogenesis, and its increased levels in spermatocytes suggest a role for BRG1 in the progression of meiosis.
Brg1 deficiency results in meiotic arrest
To determine whether BRG1 is required for spermatogenesis, we used a germ-cell-specific Cre recombinase to delete a Brg1 floxed allele (Sumi-Ichinose et al., 1997) during embryonic germ cell development. Mhv-Cre (Gallardo et al., 2007) is expressed in primordial germ cells beginning at embryonic day 15.5. We confirmed the timing and specificity of Mvh-Cre using the R26R reporter gene (Soriano, 1997) (supplementary material Fig. S2). Morphological analysis of adult Mvh-Cre/+; Brg1fl/Δ (referred to as Brg1CKO) testes demonstrated a dramatic decrease in size relative to wild-type testes (Fig. 2A). Mutant seminiferous tubules contained spermatogonial cells and spermatocytes, but lacked fully differentiated cells such as round and elongated spermatids (Fig. 2B-E). In contrast to the robust BRG1 immunostaining in the testes of wild-type (Brg1fl/Δ) males lacking the Cre recombinase, testes of Brg1CKO males showed BRG1 expression excluded from germ cells identified by expression of TRA98 – a pan-germ cell marker (Tanaka et al., 1997) (Fig. 2F,G). BRG1 was detected in somatic, TRA98-negative Sertoli cells (arrows in Fig. 2G), which do not express Cre, further demonstrating the specificity of Mvh-Cre directed excision. These results demonstrate that germ cells depleted for BRG1 are unable to complete spermatogenesis, and suggest that BRG1 is required for progression through meiosis.
To determine the cause of the arrest, we characterized spermatogenesis in Brg1CKO mice at 7 dpp when testes are populated only by spermatogonia. Compared with littermate controls, mutant testes appeared to contain normal numbers of germ cells, as marked by TRA98 (supplementary material Fig. S3A,B). Moreover, the frequency of proliferating cells, marked by phospho-histone H3 (phosphorylated serine 10, histone H3), was also comparable (supplementary material Fig. S3C,D). Thus, Brg1 appears dispensable for viability and proliferation of spermatogonia. We asked next whether differentiation of spermatogonia is affected in mutant testes. We observed comparable expression of the spermatocyte marker MVH (DDX4 – Mouse Genome Informatics) (Tanaka et al., 2000) (Fig. 2H,I) in 1-month-old testes from Brg1CKO and control testes, indicating that mutant spermatocytes can initiate differentiation.
As mutant tubules lacked germ cells beyond the spermatocyte stage, we determined whether mutant spermatocytes were undergoing apoptosis. Using a TUNEL assay, we detected increased cell death in mutant testes compared with wild-type littermates (Fig. 2J,K). Some of the TUNEL-positive cells also stained positively for the spermatocyte marker SYCP3 (Fig. 2L). Therefore, deletion of Brg1 impedes progression of spermatogenesis during meiotic stages, resulting in increased apoptosis.
Synapsis is impaired in Brg1CKO spermatocytes
To determine whether mutant spermatocytes progress properly through prophase I, we analyzed the expression and localization patterns of meiotic markers. γH2AX levels are known to increase during leptonema, correlating with DSB formation, then diminish as breaks are repaired during zygonema, eventually becoming confined to the XY bivalent by the pachytene stage (Mahadevaiah et al., 2001; Celeste et al., 2002; Fernandez-Capetillo et al., 2003). We observed a pattern of γH2AX staining in Brg1CKO tubules consistent with the leptotene stage of the first meiotic prophase. However, in contrast to wild-type tubules, where the normal pattern of the diffusely staining γH2AX in leptotene or zygotene spermatocytes is a single layer, we observed these cells accumulating in multiple layers in Brg1CKO testes (Fig. 3A,B). We next assayed SYCP3, which is highly expressed in zygotene spermatocytes, becoming confined to the axes of paired chromosomes during pachynema. Similar to γH2AX staining, intensely staining SYCP3 spermatocytes accumulated abnormally in Brg1CKO tubules (Fig. 3C,D).
To examine the defect more closely, we monitored meiotic progression by using spermatocyte spreads immunostained with markers for axial and transverse elements of the synaptonemal complex, SYCP3 and SYCP1, respectively. In wild-type spermatocytes, SYCP3 localization to chromosomal axes begins in the leptotene stage, continues along the aligning axes in the zygotene stage, and forms a condensed synaptonemal complex by the pachytene stage (Fig. 4A). Although SYCP3 patterns reflecting zygonema and pachynema could be detected in Brg1CKO spermatocytes, localization of SYCP3 to the synaptonemal complex was abnormal (arrows in Fig. 4B). A significant increase in pachytene nuclei displaying asynapsis (70/80 nuclei) was observed in mutants compared with wild type (29/127 nuclei) (P<0.001, two-sided Fisher's exact test). Examination of SYCP1 localization, which becomes enriched at the transverse element bridging the two homologs together, confirmed that Brg1CKO pachytene spermatocytes fail to produce a complete synaptonemal complex (compare Fig. 4C, wild type, with 4D, mutant, incomplete synaptonemal complex highlighted with arrows). Furthermore, after quantifying the stages of meiosis I from nuclear spreads, we did not observe any diplotene stage spermatocytes at 20 dpp in Brg1CKO tubules (supplementary material Fig. S4). Together these observations demonstrate that synapsis is impaired in the absence of BRG1.
DNA repair is compromised in Brg1CKO spermatocytes
A defect in processing of DSBs is often associated with abnormal synapsis (Baker et al., 1995; Burgoyne et al., 2009). BRG1-depleted zygotene spermatocytes consistently displayed prolonged γH2AX expression (Fig. 4B,D), implying that unrepaired DSBs are persistent and that deficiency in DNA repair might be linked to incomplete synapsis. Therefore, we used immunofluorescence to analyze levels and distribution of RAD51 (Shinohara et al., 1993) and replication protein A (RPA) (Plug et al., 1997), both of which are critical components of the DNA repair complex. RPA binds single-stranded DNA, and it is replaced by RAD51, which facilitates DNA strand exchange. Both factors showed irregular distribution and kinetics in mutant spermatocytes (Fig. 5A-D). Each formed enlarged and persistent foci between zygonema and pachynema, suggesting abnormal repair of DSBs (Petrini and Stracker, 2003). Notably, the exacerbated foci were associated with increasing synaptic defects (arrows in Fig. 5B,D). The majority of pachytene spermatocytes (58/79, 73%) contained multiple, enlarged RPA-foci accumulating to locally asynaptic axes. These patterns were never observed in wild-type spermatocytes.
In mouse, ∼300 DSBs per leptotene nucleus are generated by SPO11 (Keeney, 2001). DNA repair generates two alternative recombination products, crossover (CO, <10% of all the DSBs) or non-crossover (NCO) (Moens et al., 2002). To compare wild-type and Brg1CKO recombination patterns, distribution of the recombination protein MLH1 was examined. MLH1 specifically marks CO-designated sites in mid to late pachytene spermatocytes (Moens et al., 2007). Although CO formation in Brg1CKO spermatocytes was readily observed, the average number of MLH1 foci in each nucleus was significantly reduced in mutant pachytene spermatocytes compared with controls (Fig. 5E-G, 17.3 versus 21.1 MLH1 foci/spermatocyte, P=2.3×10–6). Mutant nuclei with fewer than 20 MLH1 foci displayed extensive asynapsis, suggesting that the decrease in MLH1 foci among mutant spermatocytes is likely to be a consequence of delayed synaptic progression, possibly linked to deficiency in DNA repair.
Brg1CKO spermatocytes show abnormal chromatin modifications
SWI/SNF chromatin remodelers such as BRG1 are thought to produce a more open chromatin structure (Bultman et al., 2006). To determine whether the failure in meiotic progression correlated with changes in chromatin structure in the absence of BRG1, we examined global patterns of histone modifications and chromatin-binding proteins. We first investigated acetylation of lysine 14 of histone H3 (H3K14Ac), a modification thought to facilitate the recruitment of DNA damage response proteins (Lee et al., 2010; Buard et al., 2009). We found that, although H3K14Ac was induced in mutant spermatocytes, it persisted abnormally into the pachytene stage (Fig. 6A,B). Similarly, dimethylation at histone H3K9 (H3K9me2), a heterochromatin-associated modification, was also abnormally increased in mutant tubules (Fig. 6C,D). Staging nuclear spreads with SYCP3 revealed that this upregulation occurred during the zygotene stage, preceding the period of ectopically increased H3K14Ac (Fig. 6E,F).
Heterochromatin protein 1 (HP1; CBX5 – Mouse Genome Informatics) is a non-histone protein that regulates the assembly of heterochromatin (Zhao et al., 2000). We examined expression of all three members of mammalian HP1 proteins: HP1α, HP1β and HP1γ. HP1α expression was negligible in both wild-type and mutant spermatocytes (data not shown). In wild-type tubules, HP1β and HP1γ strongly stained cells at the base of the lamina with levels decreasing as cells entered meiosis and moved towards the lumen. Only HP1β was reactivated in pachytene spermatocytes (Fig. 6G). In Brg1CKO tubules, broad staining of both HP1β and HP1γ persisted into spermatocytes (arrows in Fig. 6H), which contained abnormally high levels of γH2AX (Fig. 6J). Taken together, these results show differences in the patterns of histone modifications in the absence of BRG1 during meiosis and suggest a more closed, heterochromatic state.
Chromatin-remodeling proteins have significant roles in facilitating DSB repair and maintaining genome stability (Downs et al., 2007). Here, we demonstrate an essential role for a catalytic subunit of the mammalian SWI/SNF complex, BRG1, during the programmed DNA damage events of meiosis in the male germline. Although DSB repair appears to begin normally in Brg1CKO spermatocytes, DNA damage persists into later stages of prophase I, as indicated by persistent γH2AX, unresolved repair foci and damage-associated H3K14Ac, suggesting that DSB repair is unable to proceed to completion. Subsequently, progression through the first meiotic prophase of Brg1CKO spermatocytes is blocked, resulting in the elimination of the spermatocytes presumably by meiotic checkpoint-triggered cell death. Recombination and synapsis are mutually interdependent, such that the resolution of recombination intermediates requires assembly of intact synaptonemal complexes (Plug et al., 1998). Unrepaired DSBs in the Brg1CKO mutants might result from incomplete synapsis, deficiency in DNA repair, or some defect upstream of synapsis/repair. Intriguingly, we observed the presence of crossover sites, as measured by MLH1 foci, in Brg1CKO mutant spermatocytes despite measurable asynapsis. It is possible, therefore, that BRG1 is specifically required for synapsis in condensed, heterochromatic domains.
Previously, Park et al demonstrated that upon induction of DSBs by ionizing radiation, BRG1 is required to promote high levels of γH2AX for efficient repair (Park et al., 2006). In contrast, we observe that γH2AX appears normally in Brg1CKO spermatocytes during the leptotene stage of meiosis. However, γH2AX distribution was abnormal. Instead of being normally displaced from autosomes and confined to the XY body, γH2AX remained broadly dispersed in mutant zygotene spermatocytes and into the pachytene stage. Therefore, during meiosis, BRG1 appears to function downstream of γH2AX accumulation. This discrepancy may reflect mechanistic differences in DNA repair that occur in response to stress and those that occur during meiosis (Andersen and Sekelsky, 2010). The timing of BRG1 requirement during meiotic DSB repair is reminiscent of that observed in yeast. In this organism, the SWI/SNF remodeling complex appears to be involved later in the DSB repair process, perhaps during or after the invasion of single-stranded DNA to donor DNA (Chai et al., 2005).
We have previously observed differences in the histone modifications of BRG1-depleted embryos (Bultman et al., 2006). Here, we show that Brg1CKO spermatocytes undergo changes in heterochromatin-associated histone modification patterns. These observations suggest a more condensed chromatin state relative to wild-type spermatocytes. Heterochromatic regions require SWI/SNF complexes to complete recombination in yeast (Sinha et al., 2009). Thus, the defect in DNA repair coupled with the increased heterochromatic nature of Brg1CKO spermatocytes suggest that mammalian SWI/SNF complexes perhaps facilitate DSB repair and recombination in more condensed genomic contexts. Together, our results suggest an essential role for the BRG1 ATPase in mammalian male meiosis.
We thank the members of the Magnuson Lab for critical reading of the manuscript; Joshua Starmer for statistical analysis; Dale A. Ramsden (UNC) for anti-Rad51 antibody; Dr D. H. Castrillon (UT Southwestern for the Mhv-Cre line, provided by Dr Blanche Capel, Duke University); and Dr Pierre Chambon (Strasbourg) for Brg1flox mice.
This work was supported by a National Institutes of Health grant [HD036655 to T.M.] and a National Research Service Award (NRSA) Fellowship [HD0052413 to A.M.F.] Deposited in PMC for release after 12 months.
Competing interests statement
The authors declare no competing financial interests.
Supplementary material
Supplementary material available online at http://dev.biologists.org/lookup/suppl/doi:10.1242/dev.073478/-/DC1
- Accepted January 9, 2012.
- © 2012.


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Dolce & Gabbana
Boys Black Wool & Gold Guitar Blazer
Boys black blazer from Dolce & Gabbana. Woven in a luxurious wool and silk blend, this smart jacket has silky satin lapels and a button-hole. There is a chest pocket and two stitched hand pockets with a gold-coloured guitar appliqué. The blazer fastens with two covered buttons at the front and there are three covered buttons at the cuffs. Inside, it is lined in a floral jacquard silky fabric, with an interior buttoned pocket and a stitched logo tag.
- Product number 154544
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- Lining: 58% viscose, 42% cupro (satin feel)
- Dry clean
- Inside pocket with button fastening
- Designer theme: Musical
- Button fastening


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Third graders create a butterfly drawing that includes the elements of art and principles of design. They create artwork that can be made into ceramic tiles to be installed around the classroom doorways in our new addition.
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Aaron Mooy
forbze replied to MHFCRC's topic in Melbourne CityAll part of the CFG master plan. It doesn't really matter if Melbourne City are making money or not because we are not under financial fair play rules - we can actually be losing 20 mil a year and it doesn't matter. If we sell Mooy to Man City for peanuts, and then loan him out for a sizable loan, it's all profit for their club - which means they are closer to profit and FFP in Europe.
Malicious Site Warning
forbze replied to silva10's topic in MetaI just got redirected just now... still busted.
Melbourne Heart FC buy out - Man City, Melbourne City FC, etc.
forbze replied to AntiScum's topic in Melbourne CityXavi set to sign a 3 year contract with NYCFC this week according Fox Sports. Xavi and Lampard on Loan... Feeding Villa
Trumpet Notes
forbze replied to Bozza's topic in ChantsPretty sure the name of the song in the Eskisehirspor vid is Espana Cani - Sheet music here: http://www.sheetmusicstock.com/sheetmusic/11129/espana-cani?r=smt
The 'what are you listening to right now?' thread
forbze replied to marteaux's topic in The Club Room
Western Sydney
forbze replied to LR9's topic in Australian FootballFFA Have a trademark pending on the names - Wanderers FC, Wolves FC, Athletic FC, Strikers FC. Could be any combination of Western Sydney, West Sydney, or Sydney West in the name.
Player Contracts - Season 2018-19
forbze replied to forbze's topic in Melbourne CityErr.. yeh, the numbers aren't shirt numbers - I just copied and pasted from the wiki!
Player Contracts - Season 2018-19
forbze posted a topic in Melbourne CityHere's all the contract details for the players who are still at the club - or we are unsure about.<p>1 Clint Bolton (End of 2012/13) http://socceranchor.com/index.php?option=com_content&view=article&id=3905:melbourne-heart-sign-youngsters-to-new-deals&catid=58:latest-news&Itemid=169"]Source 4 Simon Colosimo (End of 2012/13) 5 Fred (End of 2012/13) 6 Matt Thompson (End of 2012/13) 8 Jonatan Germano (End of 2012/13) 9 Eli Babalj (End of 2012/13) http://socceranchor.com/index.php?option=com_content&view=article&id=3905:melbourne-heart-sign-youngsters-to-new-deals&catid=58:latest-news&Itemid=169"]Source 12 Jason Hoffman (End of 2012/13) [url="
Membership numbers and how to increase them
forbze replied to Torn Asunder's topic in Melbourne CityIs there a Operation 12,000 thread?
Heart Announces Cuts to Squad
forbze replied to ToonHeart's topic in Melbourne CityI think I read somewhere once - "If they didn't win the championship for you this year... they won't next year". Unfortunately that is the case for Alex Terra and Worm. They're at an age now, where they aren't going to increase in ability, so really were just taking up a spot that could be filled by a future star from the youth team, or brought in player of higher quality. Alex gave us all a couple of special moments, and I'm sure every kid in the crowd will be asking where he is next year, but I think this was expected...
AVB and Orjan Nilsen just released a Belter...
Prob a bit too chilled for trance - but this is awesome.
Membership numbers and how to increase them
forbze replied to Torn Asunder's topic in Melbourne CityRe-Paint pedestrian crossings red and white stripes all over the city with a stenciled heart logo. current membership level is just over 7000


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Best of CCTV | CCTV English - October 3, 2017
Smash hit: Ping pong robot takes on Olympian at Tokyo tech fair - October 3, 2017
Trump says Las Vegas shooting an 'act of pure evil' - October 3, 2017
Witnesses described the gunman Stephen Paddock opened fire - October 3, 2017
White House calls Dem demands for action on guns 'premature' - October 3, 2017
Las Vegas shooter perplexes law enforcement - October 3, 2017
Las Vegas shooting: At least 59 dead at Mandalay Bay Hotel - October 3, 2017
Exclusive interview with Dr. Jing Yin – What is the biggest challenge for Buddhist today? - October 3, 2017
How a novel was adapted into a movie : Interview with Yan Geling - October 3, 2017
Red Flowers All Over The Hills - October 3, 2017
China's first nuclear plant in ethnic minority region begins operation
The No. 1 reactor of the Fangchenggang Nuclear Power Plant in Guangxi Zhuang Autonomous Region began commercial operation on Friday.
It's China's first nuclear power plant in the ethnic minority regions.
The reactor can supply 24 million kilowatt-hours of electricity a day, which could meet the needs of a medium-sized city.
The No.1 reactor first began generating power in October 2015. The plant's No. 2 reactor is expected to start operating in the second half of 2016.
SOUNDBITE (CHINESE): GONG GUANGCHEN, Spokesman, Fangchenggang Nuclear Power Plant
"The two reactors can create some one million new jobs annually and are expected to contribute 8 billion yuan to the local GDP."
Gong says the reactors will cut coal burning by 4.8 million tonnes and CO2 emissions by about 11.9 million tonnes every year.
China has seen rapid nuclear power growth in recent years, but it suspended approval of new programs after Japan's Fukushima nuclear disaster in 2011.
It was not until 2014 that the country restarted nuclear power plant construction.


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E1: SVM: Server Manager Agents Sporadically Have Connectivity Conflicts Causing Agents To Go In Wait Status, Hang, And Be Non-Responsive
Last updated on SEPTEMBER 08, 2016
Applies to:JD Edwards EnterpriseOne Tools - Version 8.98 and later
Information in this document applies to any platform.
Server Manager will be running fine. Intermittently Server Manager Agents hang, They do NOT all hang at the same time. But, it can happen to any SM agent.
You may have gone to new hardware or made changes to your hardware immediately before this problem began to occur. This is a sporadic problem. It may be difficult to isolate the exact time it began to occur.
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Deep House, Tropical House Gurre
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Mold Technician
- Employer
- Dunlop Protective Footwear
- Location
- Havre De Grace, MD
- Posted
- May 19, 2017
- Closes
- May 22, 2017
- Function
- Accountant, IT
- Industry
- Specialty Trades
- Career Level
- Experienced (Non-Manager)
- Hours
- Full Time
Job DescriptionBuilds and repairs machine shop tools using machining operations and measuring instruments. Works from blueprints and sketches to create or modify tools, dies, jigs and molds. Requires a high school diploma or its equivalent. May be required to complete an apprenticeship and/or formal training in area of specialty with 2-5 years of experience in the field or in a related area. Familiar with standard concepts, practices, and procedures within a particular field. Relies on experience and judgment to plan and accomplish goals. Performs a variety of tasks. Works under general supervision. Typically reports to a supervisor or manager.Company DescriptionDunlop Protective Footwear is the global market leader in protective footwear for the boots segment. Having an excellent brand name and being renowned the world over with our Purofort products, we design, develop and manufacture all of our high quality waterproof boots ourselves, combining original expertise with new technologies. We offer our customers worldwide with the highest level of safety footwear for professional use in segments such as Agriculture, Food Processing, General Industry and Oil & Gas.We have recently acquired the former A' oeONGUARD IndustriesA' , which will serve as our USA Business Unit to further extend our market reach. Onguard has been a leader in this market of protective boots for many years and with the new Dunlop brand equity, its innovative Purofort line of products and marketing resources we see ample room for growth. We are looking to extend our market position in the USA by extending the business in new market segments and a wider product range being available.


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Tribal communities in Odisha to boycott twin polls
People belonging to five tribal communities in Koraput district in Odisha have decided to boycott assembly and Lok Sabha polls
Koraput (Odisha): People belonging to five tribal communities in Koraput district in Odisha have decided to boycott assembly and Lok Sabha polls, protesting against the government's failure to give them tribal status.
Leaders of Dora, Muka Dora, Nuka Dora, Rana and Mali communities under the banner, Koraput Zilla Bana Basi Maha Sangh (KZBBS), said the decision was taken over the weekend.
"Around two lakh voters of our communities will boycott the upcoming twin polls. Many times we have approached both the state and Union governments regarding our demand, but in vain," Secretary of Sangh, Trinath Gamel said.
According to KZBBS president Gokul Chandra Khoda, "Members of the communities are deprived of various benefits meant for tribals. Government should include us in the list of tribals and launch a special project for our development."
People of these communities mostly reside at Pottangi, Semiliguda and Nandapur blocks of the district and make a living for themselves by undertaking minor cultivation and collecting minor forest produce.
Assistant sub-collector (Koraput) Muralidhar Swain said, "It is up to the community members to decide whether they should participate in the elections or not. We cannot force anybody to take part in poll process."


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All right, I can do this. Back on the "A-to-Z" thing.
I just saw some news over at The Other Side blog that Margaret Weis Productions will no longer be making books for Marvel Heroic Roleplaying. Tim, the blogger at The Other Side, speculated that it was because you needed the right group to make it work. I've been playing it a few times with my group, and he might be right.
I'll also go one further and say that the game's lack of rules for building your own character and advancing that character, both staples of most RPG's, probably didn't help.
I own the core rulebook and the Civil War supplement in hardback. (I'm not a fan of pdf's for reasons I'll go into on another day.) But I pre-ordered The Initiative through my FLGS, and now they won't be releasing the book. Here's the news release, by the way.
Naturally, not having a game in print doesn't mean you can't play it, and I'll probably continue on with MHR for a while, at least until its novelty wears off. I have noticed a bit of fan backlash about this, by the way, mostly on the grounds of the pre-orders that won't be fulfilled and that MWP got in bed with Marvel, which hasn't really had a successful RPG license since FASERIP, and whose expectations may have been ridiculously high.
In other, non-supers related news, I've been hanging around today working an a fantasy RPG project that'll probably keep me occupied for a few weeks, in anticipation of getting both my Blade Raiders game and my Reaper Kickstarter package. I'll post some news about it if I can make a little headway on it first. I wouldn't want to start something without being able to finish it... Oh wait, it's still April and I'm only on "U."
Friday, April 26, 2013
U is for Underperforming Supers RPG's
3 comments:
I recently bought the Marvel RPG for my daughter, who is in college. It is disappointing to hear that MWP will not be continuing with it. My daughter and I really liked the Supernatural RPG, another MWP product.ReplyDeleteReplies
Apparently they can't sell anything after the end of the month. Their pdf's are really overpriced in my mind, so I will probably be taking a pass.Delete
I think the main problems with the MWP game were (1) it was poorly written to the point of being confusing--it's the ONLY game I gave up trying to learn, and (2) the modules were simply reenactments of crappy storylines published a few years earlier--if you've read the stories, there's no suspense or surprise.ReplyDelete
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One thing I've been slowly working on for the last year is another fantasy sandbox campaign. My prior one was generally map-based, alth...
For a long time I've been in the market for a new supers RPG. Since running Marvel Heroic Roleplaying a few years ago, I've been l...
The Red Hood's gang of re-furbished C-listers The general consensus among my kids is that they want me to continue to run a superher...


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Beaver Hunt #5
Hustler Video
Genre: Gonzo / Vignette
Condoms: None
Length: Slightly over 2 hours
Director: Will Rider
Date of Production: 6/8/06
Cast: Riley Shy (cover), Siena Moore, Piper Parker, Ryann Reynolds, & Addison Rose
Special Features: As with the previous installment in the series you get a decent, but not great, selection of extras. The best is a 32 minute behind the scenes featurette. It sheds a little light on what it takes to actually make an adult DVD. The other extras include a Slide Show, Trailers, and Sex Talk commercials.
Audio/Video Quality: As is the case with most Hustler releases, this title is presented in a 4:3 full screen format. Shot with a handheld digital camera, the lighting was almost always solid, resulting in excellent crisp colors, lifelike flesh tones, and minimal shadows. There were no compression artifacts or pixilation on the print, just as you'd expect from something shot by a high-end studio. The stereo 2.0 audio was pretty basic with the actors being heard easily enough but there was little separation between the channels and it had a limited dynamic range. Overall it was another solid job of Hustler.
Scene 1 - Addison. Addison is an incredible looking girl; she really looks like she could attend your local college. After a quick interview and some seductive masturbation, Addison meets her partner. She wastes little time before diving down and slumping up is cock. Despite the rough treatment (face fucking), Addison always looked like she was enjoying it. They begin fucking in the doggy style position and go in to missionary and cowgirl. After nearly twenty minutes, Addison gets a load of jizz sprayed on her face and chin. Solid opening scene.
Scene 2 – Riley. Riley is the brunette cutie on the cover of the DVD. After her introduction, Riley spends several minutes getting fingered by her man. She returns the favor by performing some very upbeat oral on him. If you think she looks good on the cover wait till you see her with a cock in her mouth. Riley gets violated in the following positions: missionary, cowgirl, reverse cowgirl, and doggy style. The scene ends wit Riley receiving a fire hose like blast of man gravy poured all over he r face. This was the first time I've seen Riley and I was blown away.
Scene 3 – Ryaan. Ryaan is an attractive black girl with a great personality. You can tell just from her interview that she'd be really fun in bed. After showing off her smooth pussy, Ryann gives her man some sloppy wet head. Despite his huge meat stick, she had no problems handling him. The action begins in the reverse cowgirl position and goes on to missionary and doggy style. A close-up of Ryaan getting a load of sperm deposited on her tongue ends the scene. A decent scene all-around with plenty of passion and chemistry.
Scene 4 – Piper. Despite having the strange name of Piper Parker (no not Spiderman), this blonde cutie might be the best looking girl on the disc. She spends a few minute splaying with herself before her partner comes in and takes over. Piper gives him a solid blowjob with all the energy and enthusiasm you could hope for. Piper gets a vicious pummeling in all your standard positions before the climactic ending. While on he knees, Piper gets blasted in the face by a massive load of man gravy. Great girl, great scene.
Scene 5 – Siena. Last, but certainly not least, is Siena. Her gorgeous face and tight young body make her quite the looker. After a quick interview, Siena greets her man with a sloppy blowjob. She really attacked his cock like she hadn't been fed in weeks. Siena then jumps his bones and starts fucking in the reverse cowgirl position. She also gets violated in doggy style, missionary, and piledriver. She swallows a weak cumshot at the end of the scene. Overall a very enjoyable scene.
Final Thoughts: I was a big fan of Beaver Hunt #5, and this release provides the same level of quality. You get five young girls who are not only smokin' hot, but they love to fuck on camera. Only the lack of a bonus scene (or something similar) prevents me from giving this DVD our highest rating. It still earns a well-deserved rating of "recommended". Now get out there and watch this DVD!
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I love how thousands and thousands of people LOVED the Parade and The Gasser Lounge entry, and one little mouse voice goes "squeak, squeak, spueak..." and gets a letter to the editor printed (below). Screw Hany! We're No. 1, We're No. 1. No time for Losers (Hany), cause We're No. 1, We're No. 1... Suck it.
And obvioulsy no problems with "alcoholism" in her selective bitching-rant, cause there was tons of it there, on the trucks (Budweiser and others were actively drinking along the whole route) so screw your selective chastity. It's 11:00 AM and "everyones drinking kids, that's perfectly ok." No one got naked, everything was done in good taste... (Have you walked a mere BLOCK to the beach and seen all the bikinis, thongs, and perky nips? Welcome to reality of living in a Beach City) good enough to... say it with me... "We're No. 1, We're No. 1!!!"


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How Many Human Rights Abuses are Acceptable?
|Not in Fiji
It is not possible to briefly answer this question to the satisfaction of the reader. What appears to be a simple request for a concrete figure (presumably '0') is in fact a loaded request to put a figure on an abstraction. I am asked to make a value judgment in the absence of common agreement on whether each HR is of equal importance; whether each is always applicable, irrespective of the circumstances; whether everyone's HRs are of equal weight and importance, irrespective of individual and communal circumstances;whether it is ever justifiable to deny some HRs to some people; and whether some HRs can justifiably be temporarily suspended for the 'common good.'
To defuse the political element (which is the obvious point) of the question, I suggest we ask the same questions of "sin" where the answers will be seen to be no less difficult.
The reader knows my general position on HRs in Fiji. I have criticised abuses when I have known them to have occurred and have recommended better practices. He also knows that the level of the alleged physical abuses have been exaggerated. Anti-Government people have referred to torture but they do not even come near to what people associate with the word "torture", as seen in Iraq, Afghanistan and on TV thrillers. And he knows I have called for the lifting of the emergency regulations.
I suspect the reader's motive for asking the question is because he wants to draw attention to HRs abuses since December 2006. But the reader must know that the abuse of basic HRs did not suddenly start in 2006; that most physical abuses occurred in the immediate aftermath of the coup; that most people are not affected (on a world scale Fiji abuses might score 5 out of 100), and that the HRs situation appears to be getting better. The reader should also know that those who complain most about the post-2006 abuses were singularly quiet about human rights abuses in 1987 and 2000-01 (presumably because they did not affect them, or their ethnnic group); and even now such people speak only of their chosen "middle class" HRs, saying nothing about social justice and the denial of basic HRs to Fiji's second class citizens. -- Crosbie Walsh


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Spin Tires Demo - Mods
What do you think about this video?
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Spin Tires Demo - Mods 5 out of 5
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Making red carpet appearances is all part of the job for celebrities.
They must spend hours in the run up to these events thinking about their outfits and accessories and ensuring they will stand out from the celebrity crowd and have a flattering review of their look in the media the following day.
On the day of the event itself the celebs must spend hours making sure they look red-carpet ready. From getting their hair done to having their makeup applied it's hours of effort for their five minutes of fame. So, from organising their dress, to their matching accessories, to their makeup and hair – it really is a full time job preparing for such events.
After spending endless hours watching the Kardashians in all their glory on their hit reality TV show 'Keeping up with the Kardashians', we have come to realise that these glam, red carpet events truly are the biggest stresses celebrities face.
Not that they should stress though, Kim K always looks amazing on the red carpet.
It is not surprising then that celebrities have their own glam squad, that take care of all their red carpet looks and make them camera ready – this is serious business. These teams travel everywhere with the celebs to make sure they are looking glam at all times.
Dressing in their best and standing beside other great and important people is just one of the ways celebrities stay in the press and how they stay relevant.
If one were to stop making public appearances, they're at risk of falling out of the public eye.
Take the Met Gala for instance, how many weeks were people talking about Beyonce after that event? I mean, she looked incredible, and even fellow celebrities commented on how good she looked!
Whereas one bad outfit can have the same effect and put you in the tabloids for weeks – something our girl Riri is used to due to her daring outfits. And this Met Gala look was nothing surprising…
Yes – Rihanna is hiding amongst those flowers! This look catapulted Rihanna into the spotlight, for the mixed opinions on this risky look. Nevertheless, as previously mentioned, it definitely made sure she was in the public eye.
And who can forget this Lady Gaga red carpet look?
She is definitely not one to shy away from crazy outfits, but this look definitely made sure she was in the spotlight for a good week atleast. Although the dress is classy, and very normal (especially for Lady Gaga) – she had to go and wear a pair of red marigolds! Some may say this made her look, and others may say it ruined it. However, one thing we can all agree with…she definitely got people talking and left an impression on people.
That's why red carpet events are a big deal for celebrities.
But there are a few tips that ALL celebrities follow to ensure they both look and feel their best when they are in front of the camera…and it is all related to their shoes!
That's right, there is detail in every single aspect of a celebrity's red carpet outfit and their shoes are no exception.
However, some reporters have recently noticed that some female celebrities are wearing shoes that are a size or two too big for their feet.
Is this a new fashion trend? Apparently not. It just appears that these fashion conscious celebs are onto a secret trick that most of us don't know about.
Many of the Hollywood stars who flaunt their outfits on the red carpet generally wear high heels. But because of the nature of the events the celebrities are wearing the heels for too long, and things can get a tad uncomfortable.
By wearing shoes that are a bit too big for their feet, they are allowing their feet to breathe. It also prevents the feet from swelling and forming blisters.
If these celebs wore shoes that were their shoe size, by the end of the night, their feet would be covered in blisters and swollen.
And how often have you seen a celebrity with cut feet due to their high heels? This is something we have actually never thought about – but it is true. Whereas we spend one hour in heels and are feet are bleeding and full of blisters!
So to avoid any foot discomfort, blister, or any sort of foot related pain, celebrities often choose to wear shoes that are at least a size too big.
Another benefit is that large shoes, pumps or sandals won't pinch, squeeze or rub the feet in the vulnerable areas of the foot (like the balls of our feet, or in between our toes).
Although most people would put a bandage on these vulnerable areas to prevent blisters, celebrities aren't always allowed this as an option since cameras are constantly pointed at them, ready to capture and wardrobe malfunctions.
Can you imagine seeing a photo of an A list celebrity wearing blister bandages!?
Now you may wonder, how do they keep the shoes on if they're a size too big?
We for one can't even keep shoes on that fit us properly, never mind a pair of shoes that are too big. And they manage to make it look so easy?!
Well, that's a great question!
They manage to keep them on because their designers usually place silicone pads inside the pumps.
This method ensures that the shoe fits snugly and rather comfortably on the foot all the while blood flow is not restricted and the feet won't be pinched…
Double-sided tape is also used in order to keep the shoe on all night long. This is similar to body tape which us ladies use all too frequently to keep our clothing tight to our chest to avoid that dreaded nip slip. Apparently this works on shoes too? This is definitely something we are going to try out.
But a simple way to avoid any weird looks could just be to wear a longer dress that can hide the shoe or to simply wear pants where the hem of the pants cover the shoes.
But should celebrities be limited to covering their feet at every red carpet event? Especially when the vast majority spend a considerable amount of time working out to get that perfect body to show off at such events.
However, this trend in wearing oversized shoes has been steadily growing so celebrities don't even need to go about hiding their oversized shoes.
So there you have it, hot Hollywood tips fresh off the press! Now you can grace your next formal event without the awful blisters, cramps, and foot pain! You're welcome!
Feel free to let us know what you think in the COMMENTS!
Don't forget to SHARE this with your friends and family so they can see it too!


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This informs users automatically if the site they are browsing has any available coupon codes, removing the need for the user to browse through codes. By alleviating this burden, fewer codes are missed and users remain more engaged with the plugin.
Alexandria, VA (PRWEB) February 28, 2014
The leading cash back plug in developers Besttoolbars have published their annual list of killer features for successful cash back plug ins. The 2014 run down recognizes the increasing speed with which the cash back marketplace is changing and advancing. It picks through the ever growing pool of possible new features to highlight the ones that have the potential to have the best impact on revenue and attract the most end users, without overloading the extension with a mass of programming extras.
The list entitled "Killer features of a cash back plugin in 2014" contains four top tips. Valery Kuznetsov, Key Projects Manager at Besttoolbars said, "The question we asked ourselves when compiling this list was, what is it that can really attracts users? Through collecting and analyzing many different reviews we concluded that in today's fast paced lives, there are two important things for users when using cash back plugins. The first is that the application should be accessible and always at hand and the second, that the process of finding great deals and making the purchase should take as little time as possible. Using these two blocks as the starting point we created our shortlist of killer features to make 2014 cash back plug ins really successful."
Heading the list of superstar features is combining a cash back plug in with a custom browser. This is based on a new tool developed by Besttoolbars which allows developers to create custom browsers on a Chromium open source browser with a range of customization and branding personalization options. This allows the developer to preset all extensions to be used and means the browser is not restricted by third party add-on policies.
The second key feature is a mobile cashback app. This feature made the list as a result of the growth of mobile computing, which looks certain to continue its year on year growth for 2014 – current estimates project that around 40% of traffic to ecommerce sites comes from mobile devices. A mobile app means those accessing from a tablet or similar can still access cash back discounts, helping to secure more revenue and higher numbers of users.
The third key item on the must have features list for 2014 is coupons auto apply functionality. This informs users automatically if the site they are browsing has any available coupon codes, removing the need for the user to browse through codes. By alleviating this burden, fewer codes are missed and users remain more engaged with the plugin.
Finally, promoted merchants are also advisable. This involves an add on menu, which was identified by Besttoolbars as a great way to promote select merchants in an unobtrusive way, reducing the search time for great offers substantially.
To find out more about Besttoolbars and to read the 2014 list in full visit http://besttoolbars.net/articles/killer_features_of_cashback_plugins_in_2014/.
About Besttoolbars: Besttoolbars is a company well known for creating browser toolbars and add-ons. It has worked with an array of global brands, from start-ups to high-profile Fortune 500 corporations including Amazon, Skype, IBM, DHL, TNT and Intel. More than 7000 companies currently use solutions created on the Besttoolbars platform. Its product line includes:
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Homemade Veggie broth is SUPER simple!!! There are so many reasons to make your own.
1 - You know exactly what's in it.
2 - It's green. You can reuse the jars over and over.
3 - It's cheaper....honestly it is.
4 - It tastes better.
Veggie broth can be used in a ton of different recipes and is detox friendly. Tastes great in place of water when you make rice or quinoa.
I don't really have a recipe, it's really more of a method.
I start with my giant canning pot and fill it full of organic veggies and scraps.
For this batch I used leeks, carrots, onions, potatoes, celery, mushrooms, parsley, marjoram, thyme & a couple cloves of garlic. I scrubbed the veggies clean with a strong bristle brush, chopped them in large chunks & added them to the pot.
My pot was about 3/4 of the way full with veggies.
I then added filtered water to about 2 inches from the top of the pot.
Bring to a rapid boil and then turn down to a simmer. Simmer for about 4-6 hours. The stock is done when it has a golden hue.
Next I use a large ladle and wire mesh strainer
and strain the broth into a pitcher. I put a small strainer on top of 1 of my jars and pour the broth into the canning jar. I double strain because the potatoes become mush and can cause your broth to be too thick.
Fill jars but make sure you leave 2 inches of headspace, especially if you plan on freezing the stock. Another note, you must use wide mouth jars to freeze your stock. I use wide mouth quart jars.
Allow your jars of stock to cool overnight in the fridge before putting in the freezer. Also, make sure to not screw the lids on too tight. All of these things will help to keep your jars from cracking in the freezer.
Another note, you could add salt to your broth for more flavor but I prefer to not have the added sodium.
This method typically makes 12-14 quart jars of stock.
You can feed the leftover mushed veggies to your chickens or add it to your compost pile.
Happy Healthy Eating Friends!


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In a fairly rare occurrence, Jazz and Smoke are sharing my chair.
Thursday, January 28, 2010
3 comments:
Well the felines do look comfy! Hootie and I share seats and beds all the time. Sometimes we look like one dog with 2 heads!!ReplyDelete
Smooches from pooches,
BabyRocketDog & Hootie
ps-The snow sculptures below look like the work of a real artist.They aren't by chance on Rancho Vista Dr are they? (Our bi-ped brother was an artist and lives there!)
Smoke and Muggles (yet another cat) often curl up very close to each other but Jazz is more stand-offish usually.ReplyDelete
No, I found the sculptures in the other end of town.
Our dogs are rarely close, unless competing for something.ReplyDelete
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More on privacy vs. public
In mulling that recent post, I realized my presumption was, "Why do people have only X number of children?" My brain completely misfired on the fact that there are those who have no children, for whatever reason.
I know a number of couples who have no children. Two divorced for problems related to infertility. One, it's medical. Others we have... lost touch with, to say it tactfully (not all--you know who you are). A summary, and I'll try to keep it to facts.
New Year's Eve, 2000-01. Dale and I got together with some other childless friends. I/we suspected I was pregnant and nursed my single white Zinfandel for several hours. We were all adults with nary a mortgage among us.
NYE 2003-04. We had two kids, whom we took to the get-together. Do you have any idea how tough it is to find a sitter on New Year's Eve? The little guy was still nursing, besides. Our kids are part of our family and we enjoy their company. It's one of the reasons we had them.
NYE 04-05. One of our hosts was ill and they thought it a bad idea to bring the kids, as Rachel was still not quite four months old. So we stayed home and the friends we see annually came to us the next day. And stayed for about an hour. Maybe two.
NYE 05-06. We didn't even hear from them. I mentioned it, vaguely disappointed. I'll admit not being too surprised.
Another couple I know without children was in the area, relatively speaking, last Christmas. They drove on Christmas night for probably an hour and change to visit us. At home. Without being quite sure where we were, and trying valiantly to call us (Rachel had turned on the phone and hidden it in the couch, making it impossible to get through).
They did not feel obligated to bring gifts (thank God!), but they did eat something (and then complimented me on my cooking). He had no problem conversing with Madeleine for a while. They stayed around while we bathed the kids and put them to bed. After the young'uns were asleep, we watched a video.
THOSE are friends.
But about the kids part... One never knows the reasons for a couple not having kids, or not having more, or continuing to have. It's just none of our business and asking is awfully nosy. I mean, (graphic alert) if she was raped by an uncle and then had a botched abortion that resulted in a hysterectomy at the age of 14, it's probably not something she wants to discuss with a grocery store clerk. Or like my cousin, who had no problem with her first pregnancy but hasn't gotten a second despite all avenues. But neither are things you discuss with near-strangers.
I recognize there are some who trumpet with pride their nth pregnancy (for shock value) or their or their spouse's sterilization. I think perhaps it's the loss of propriety in our society. We don't remember there are subjects not discussed in public, inappropriate clothing in certain situations, even places not to be on one's cell phone. We've lost our manners.
And that, my friends, is a whole other post.
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