text:
Rashida Jones Tweets About Shazam
Our Favorite Funny-Lady Tweets of the Week
We're rolling into August with our favorite funny-lady tweets of the week! This week on Twitter, Rashida Jones had a groundbreaking idea for those acquaintances you can't seem to remember, and Mindy Kaling shared a great tip for the professional work environment. Diablo Cody and Mae Whitman, on the other hand, focused on bodily health and willpower.
Life would be so much easier if Rashida Jones was an inventor.
I wish you could Shazam someone's face— Rashida Jones (@iamrashidajones) July 31, 2013
And she didn't even ask if we wanted any of it. Thanks Mae.
Oops ate a burrito and two tacos already today oops bye— mae whitman (@maebirdwing) July 29, 2013
Listen, we're not going to tell you how to live, Lena, but we'd appreciate some consistency.
My instagram makes my life seem full and rich while my twitter paints a far sadder picture #selfawareness— Lena Dunham (@lenadunham) July 31, 2013
Check out the rest of the funniest tweets when you read on!
Am I the only one who would be down if Mindy Kaling said this to me?
"Kiss me on the lips or you're fired!" is something you shouldn't say to your crew, I learned today— Mindy Kaling (@mindykaling) July 31, 2013
Diablo Cody sets the standard for willpower. So impressive.
I made it until 11:22 AM without cookies.
— Diablo Cody (@diablocody) July 29, 2013
Ellen Page sure knows how to appreciate nature's gifts.
A bird pooped on me!
#nature #life #blessed
— Ellen Page (@EllenPage) August 1, 2013
Something tells me Sarah Silverman is a little confused about POWs.
I feel like I'd be a great POW - only caveat is I can't bear discomfort
— Sarah Silverman (@SarahKSilverman) August 1, 2013
Oh man, to be a fly on the wall during Chrissy Teigen's therapy sessions.
Stop telling me to get a therapist because I know for a fact they would tell me to quit twitter then we all lose
— christine teigen (@chrissyteigen) July 30, 2013
Zing! Point for Bette Midler.
Kate Middleton says that breastfeeding little Prince George is more comfortable if she takes the silver spoon out of his mouth.
— Bette Midler (@BetteMidler) August 1, 2013
So that's why Anna Kendrick sounded sexier than usual.
Yes, you caught me! My allergies were giving me that super sexy voice on #SYTYCD last night. (Nasal 8 year old boys sound sexy right?)
— Anna Kendrick (@AnnaKendrick47) July 31, 2013
"You guys, we have to do it this way. It was comedy writer Megan Amram's dying wish."
Face down, ass up, that's the way I want my open casket funeral
— Megan Amram (@meganamram) July 31, 2013
Not to hate on Kristen Schaal, but I'd like to be considered for this role as well.
I could excel at any role where the character is described as chronically sleepy.
— Kristen Schaal (@kristenschaaled) July 31, 2013
Don't let Bert the Muppet hear you say that, Kristen. He'll be pissed.
Nobody is named Ernie anymore, huh?
— Kristen Bell (@IMKristenBell) July 30, 2013
I can relate to comedian Kelly Oxford. One time I got weepy from watching 30 Rock.
Just welled up with tears listening to Cheers opening theme song in case you didn't know what PMS feels like.
— kelly oxford (@kellyoxford) July 30, 2013


text:
The principal at Paul Revere Elementary School in Bernal Heights pleaded not guilty Tuesday to isdemeanor DUI and hit-and-run charges after allegedly sideswiping two vehicles just south of the Golden Gate Bridge on May 12.
Sheila Milosky, a Mill Valley resident, did not attend Tuesday's court hearing. Her attorney, Sam Ware, appeared on her behalf.
"If Ms. Milosky made a mistake here, she's going to own up to it," Ware said after the hearing, adding that he had only just received the prosecution's evidence against his client. "If she did not make a mistake and some of the facts were not reported correctly, then we're going to dispute it."
Meanwhile, Milosky still has a valid driver's license, Ware said, pending an upcoming hearing with the state's Department of Motor Vehicles.
The principal also is scheduled to appear June 26 for a pretrial conference on the DUI and hit-and-run charges.
The incident unfolded about 12:30 a.m. May 12, according to the California Highway Patrol, when Milosky was driving a white 2011 Dodge Avenger with Colorado license plates that allegedly struck a Hyundai Tucson and a stretch limousine just south of the Golden Gate Bridge toll plaza.
Milosky did not stop and was pulled over at Vista Point just north of the bridge, the CHP said. She allegedly failed sobriety tests.
Hired nearly two years ago to oversee federally mandated reforms at Paul Revere Elementary, Milosky was criticized last fall by parents who said she deployed corporal punishment tactics, including restrictive bathroom policies and silent lunches and recesses in the school's basement. The San Francisco Unified School District and other parents supported her.
Bay Area NewsCrimeCrime & CourtsLocalSan Francisco
Find out more at www.sfexaminer.com/join/

subset: commoncrawl


text:
On October 9, health professions students from the Medical University of South Carolina, the University of South Carolina, the University of South Carolina School of Medicine – Greenville, and the Edward Via College of Osteopathic Medicine - Carolinas Campus were invited to attend South Carolina's recognition of "Corps Community Day." A presentation was held on the campus of the Medical University of South Carolina and faculty and students at the other institutions were able to participate via videoconference. This program provided information about the National Health Service Corps (NHSC), which exists to increase access to primary care where it is needed most. The NHSC is a federal program that provides scholarships or repays the educational loans of healthcare providers who agree to serve a minimum of two years in a federally designated health professional shortage area.
South Carolina AHEC Executive Director David
Garr, M.D., served as moderator for the program and shared his experiences
practicing in a NHSC site in rural Utah.
Director of the South Carolina Office of Primary Care Mark Jordan provided
an overview of the NHSC and shared information on scholarship and loan
repayment opportunities. Next, attendees
viewed four short videos that showcased NHSC member stories. The videos generated awareness about the NHSC
and demonstrated the contributions NHSC clinicians are making in their
communities. Sean Boynes, DMD, one of
the dedicated professionals featured in the videos, attended the event and discussed
the benefits, working environment, gratification, and experiences he has had
working at CareSouth Carolina, a NHSC site.
The program was sponsored by the South
Carolina Area Health Education Consortium (AHEC), the South Carolina Office of
Rural Health, and the South Carolina Primary Health Care Association. For additional information on the NHSC, please visit the
NHSC web page http://nhsc.hrsa.gov/ or contact Mark Jordan, NHSC state
coordinator, at (803)898-0766 or firstname.lastname@example.com. The presentation is available online at http://news.scahec.net/2014/10/national-health-service-corps-community.html.


text:
Home > Brands > Podere Copellaro
Our mission is to bring genuine products to your tables, products that still have the flavor of the past. That's why we do not use additives or preservatives, only natural ingredients such as salt, vinegar and oil. The locally grown ingredients come from controlled cultivations, for which chemical fertilizers or pesticides are not used, and that are processed according to the recipes of the family tradition, with methods still largely handmade.
That's why our pickles and creams are high-quality, delicious in their simplicity, perfect for appetizers, entrées and side dishes to match and reinterpret at will, in order to create new tasty recipes.
Podere Copellaro offers you a wide choice of vegetables preserved in oil and in water and salt, perfect to serve as entrées, appetizers and side dishes, along with cold cuts, cheeses and accompanied by red or white wines of medium structure, or to be reinvented as one prefers.


text:
This privacy policy governs your use of the app Bathukamma Paatalu (Bathukamma Songs and Lyrics) (the "Application") that was created by Amala M (Sanketika Developers) ("us", "we", or "our").
This Application is a collection of Audio songs and Lyrics for the celebratation of the Indian festival, Bathukamma.
Data Collection by the ApplicationThe Application does not collect or transmit any data such as user name, address, phone number, email address or geographic location.
Permissions Requested by the ApplicationThe Application needs the permission to read phone state to know if there is an incoming call while playing a song. It pauses the playback of the song when there is an incoming call and resumes the playback once the call is completed. The Application does not store or transmit any information regarding the call.
Data Collection by Third Party ServicesAdMob is used to display advertisements in the form of banners and other advertisements in the Application.
AdMob is an advertising service provided by AdMob Google Inc. AdMob may display ads tailored to the User's interests and behavior, including those detected outside the Application.
For more information, please check it's privacy policy.
UpdatesOur privacy policy may change from time to time for any reason. If we make any material changes to our policies, we will mention them here in the updates section.
This privacy policy was created on March 11, 2017.
Developer name was updated on September 25, 2017.


text:
A.G.O. publishes articles that concern ALL of Henderson County
A.G.O. meets the last Thurs. of each
month 6 PM
at the First Methodist Church on Lover's Lane, in Athens.
Membership $15.00 per year for mailed journals.
Membership dues can be mailed to:
A.G.O. Treasurer
c/o Henderson Co. Library
121 S. Prairieville St.
Athens, TX 75751
2004 - 2005 Officers TX A.G.O.
Click on the names to view photos
front l - r:
La Vern Rodgers, Treasurer
Peggy Allen, Editor
Charles M. Sanders Jr., Book Committee
Back Row l - r
Bunny Freeman, website
Bennie Rust, Corresponding Sec.
Mildred Rust, Vice President
Tawna Walden, Secretary
Jana Shumate, President
A few members of TX A.G.O.:
front l - r:
La Vern Williams Rodgers, Jessica Allen, Peggy Trammell Allen,
Charles M. Sanders Jr.
Back Row l - r
Irene Garner, Mildred Allen Rust, Joyce LeMay Loden,
Tawna Loden Walden, Jana Williams Shumate, Bennie Rust,
Samuel Blair Sr., Bunny Shumate Freeman
Athens Genealogical Organization
The information
below are out
of print Journals,
Copies can be found at the Clint
Murchison Library in Athens
|Henderson Co., Texas - CSA 20th Texas Calvary
|Henderson County TXGenWeb
|Athens Genealogical Organization
|Henderson County, Texas Cemetery Records
|Henderson County, Texas - Vital Records
|Henderson Co. Census Records
|TSLAC: Genealogy
Henderson County TXGenWeb Project
This site is owned and maintained by the Henderson Co. TXGenWeb Co. Coodinator. Contents of this site have been gathered from many sources and transcribed records. Therefore, errors may occur. When in doubt as to the accuracy of data contained herein, go to the actual records yourself. The information contained in this website is for your personal use only. All pages, compilations, transcriptions and abstracts are protected by copyright law and may not be published in whole or in part without written consent of the author, contributor and/or webmaster.
© Copyright 2001 - Present Bunny Freeman


text:
The revolution you dream of is not ours. You don't want to change the world, you want to blow it up.JEAN-PAUL SARTRE, Dirty Hands
A people contending for life and liberty are seldom disposed to look with a favorable eye upon either men or measures whose passions, interests or consequences will clash with those inestimable objects.GEORGE WASHINGTON, letter to General Thomas, July 23, 1775
Revolutions have never lightened the burden of tyranny. They have only shifted it to another shoulder.GEORGE BERNARD SHAW, Man and Superman
It's the well-behaved children ... that make the best revolutionaries. They don't say a word, they don't hide under the table, they eat only one piece of chocolate at a time. But later on they make society pay dearly. JEAN-PAUL SARTRE, Dirty Hands
The tree of liberty must be refreshed from time to time with the blood of patriots and tyrants. It is its natural manure. THOMAS JEFFERSON, letter to W.S. Smith, Nov. 13, 1787
Make the Revolution a parent of settlement, and not a nursery of future revolutions.EDMUND BURKE, Reflections on the Revolution in FranceThat humanity and sincerity which dispose men to resist injustice and tyranny render them unfit to cope with the cunning and power of those who are opposed to them. The friends of liberty trust to the professions of others because they are themselves sincere, and endeavour to secure the public good with the least possible hurt to its enemies, who have no regard to anything but their own unprincipled ends, and stick at nothing to accomplish them.
WILLIAM HAZLITT, Characters of Shakespeare's Plays
Do people demand a really just system? Well, we'll arrange it so that they'll be satisfied with one that's a little less unjust ... They want a revolution, and we'll give them reforms -- lots of reforms; we'll drown them in reforms. Or rather, we'll drown them in promises of reforms, because we'll never give them real ones either!!DARIO FO, Accidental Death of an Anarchist
If we behave like those on the other side, then we are the other side. Instead of changing the world, all we'll achieve is a reflection of the one we want to destroy. JEAN GENET, The Balcony
A little rebellion now and then is a good thing and as necessary in the political world as storms in the physical. THOMAS JEFFERSON, letter to James Madison, Jan. 30, 1787
I can imagine no man who will look with more horror on the End than a conscientious revolutionary who has, in a sense sincerely, been justifying cruelties and injustices inflicted on millions of his contemporaries by the benefits which he hopes to confer on future generations: generations who, as one terrible moment now reveals to him, were never going to exist. Then he will see the massacres, the faked trials, the deportations, to be all ineffaceably real, an essential part, his part, in the drama that has just ended: while the future Utopia had never been anything but a fantasy. C.S. LEWIS, The World's Last Night
They never fail who die
In a great cause: the block may soak their gore:
Their heads may sodden in the sun; their limbs
Be strung to city gates and castle walls—
But still their Spirit walks abroad. Though years
Elapse, and others share as dark a doom,
They but augment the deep and sweeping thoughts
Which overpower all others, and conduct
The world at last to Freedom.
LORD BYRON, Marino Faliero
The right to rebellion is the right to seek a higher rule, and not to wander in mere lawlessness.GEORGE ELIOT, Felix Holt
Resistance to improvement contradicts the noblest instincts of the race. It begets its opposite. The fanaticism of reform is only the raging of the accumulated waters caused by the obstructions which an ultra conservatism has thrown across the stream of progress; and revolution itself is but the sudden overwhelming and sweeping away of impediments that should have been seasonably removed.HORACE MANN, Thoughts
The history of the human race always has been, and most likely always will be, that of evolution and revolution.LEWIS F. KORNS, Thoughts
It is far more easy to pull down, than to build up, and to destroy, than to preserve. Revolutions have on this account been falsely supposed to be fertile of great talent; as the dregs rise to the top, during a fermentation, and the lightest things are carried highest by the whirlwind. And the practice of this proposition bears out the theory; for demagogues have succeeded tolerably well in making ruins; but the moment they begin to build anew from the materials that they have overthrown, they have often been uselessly employed with regard to others, and more often dangerously with regard to themselves. CHARLES CALEB COLTON, Lacon
A constitution imperiled justifies revolution.EDWARD COUNSEL, Maxims
As to the history of the revolution, my ideas may be peculiar, perhaps singular. What do we mean by the Revolution? The war? That was no part of the revolution; it was only an effect and consequence of it. The revolution was in the minds of the people, and this was effected ... before a drop of blood was shed. JOHN ADAMS, letter to Thomas Jefferson, Aug. 24, 1815
If we would trace our descents, we should find all slaves to come from princes and all princes from slaves: But fortune has turned all things topsy-turvy, in a long story of revolutions.WELLINS CALCOTT, Thoughts Moral and Divine
Revolution does not insure progress. You may overturn thrones, but what proof that anything better will grow upon the soil? E.H. CHAPIN, Living Words
Some men hope for revolution but when you revolt and set up your new government you find your new government is still the same old Papa, he has only put on a cardboard mask.CHARLES BUKOWSKI, Notes of a Dirty Old Man
© 2012, Gabriel Thy. All rights reserved.


text:
Clinical management of abdominal aortic aneurysm (AAA) is currently limited to elective surgical repair because an effective pharmacotherapy is still awaited. Inhibition of histone deacetylase (HDAC) activity could be a promising therapeutic option in cardiovascular diseases. We aimed to characterise HDAC expression in human AAA and to evaluate the therapeutic potential of class I and IIa HDAC inhibitors in the AAA model of angiotensin II (Ang II)-infused apolipoprotein-E-deficient (ApoE−/−) mice. Real-time PCR, western blot and immunohistochemistry evidenced an increased expression of HDACs 1, 2 (both class I), 4 and 7 (both class IIa) in abdominal aorta samples from patients undergoing AAA open repair (n=22) compared with those from donors (n=14). Aortic aneurysms from Ang-II-infused ApoE−/− mice exhibited a similar HDAC expression profile. In these animals, treatment with a class I HDAC inhibitor (MS-275) or a class IIa inhibitor (MC-1568) improved survival, reduced the incidence and severity of AAA and limited aneurysmal expansion evaluated by Doppler ultrasonography. These beneficial effects were more potent in MC-1568-treated mice. The disorganisation of elastin and collagen fibres and lymphocyte and macrophage infiltration were effectively reduced by both inhibitors. Additionally, HDAC inhibition attenuated the exacerbated expression of pro-inflammatory markers and the increase in metalloproteinase-2 and -9 activity induced by Ang II in this model. Therefore, our data evidence that HDAC expression is deregulated in human AAA and that class-selective HDAC inhibitors limit aneurysm expansion in an AAA mouse model. New-generation HDAC inhibitors represent a promising therapeutic approach to overcome human aneurysm progression.
Abdominal aortic aneurysm (AAA) is a degenerative vascular disease with high morbidity and mortality, and is common in elderly people. This disease affects up to 8-10% of men over the age of 65 years and its prevalence is predicted to increase in parallel with the global ageing population (Nordon et al., 2011; Weintraub, 2009). AAA rupture is the most severe consequence of this disease, which is among the 15 leading causes of death for people aged 60-84 years in the USA (Weintraub, 2009). Commonly, this is an asymptomatic disease characterised by an irreversible degradation of the vascular wall that results in tissue failure and progressive aortic dilatation. Surgical repair is currently the only therapeutic strategy for AAA, but is associated with a substantial perioperative risk (Powell and Brady, 2004).
AAA is characterised by chronic inflammation of the aortic wall that is associated with upregulation of metalloproteinases (MMPs), depletion of vascular smooth muscle cells (VSMCs) by apoptosis and enhanced neovascularisation (Golledge et al., 2006; Forsdahl et al., 2009; Camacho et al., 2013). The mechanisms underlying AAA expansion and rupture are complex and a better understanding of the processes involved in AAA progression is crucial for the development of new therapeutic strategies. Although numerous pharmacological interventions have been proposed to limit AAA growth, none of them have provided convincing results in clinical trials (Samson, 2012; Shiraya et al., 2009; Dodd and Spence, 2011).
Recent evidence highlights the importance of epigenetics in the development of cardiovascular diseases. Among epigenetic mechanisms, those governed by histone deacetylases (HDACs) strongly affect gene transcription (Narlikar et al., 2002; Hildmann et al., 2007). HDACs constitute a family of 18 molecules divided into four classes [I, II (comprising class IIa and IIb), III, IV]. Class I and IIa HDACs regulate the expression of genes involved in key events in AAA, including VSMC differentiation, contractility, proliferation, inflammation and extracellular matrix deposition (Pons et al., 2009). Interestingly, HDAC inhibitors have been proven effective in several types of cancer (Leoni et al., 2002; Bolden et al., 2006; Minucci and Pelicci, 2006) and represent a promising therapy for non-oncological diseases, including neurodegeneration, inflammation and cardiovascular diseases (Pons et al., 2009; Colussi et al., 2010). However, the relevance of the different HDACs in human AAA has not been established. Based on the aforementioned data, we hypothesised that HDACs could be crucial in human AAA and that novel class-selective HDAC inhibitors could bring new therapeutic opportunities for AAA.
Class I and II HDACs are augmented in human AAA and in the Ang-II-infused ApoE−/− mouse model
The expression pattern of class I and class IIa HDACs was analysed in abdominal aorta samples from AAA patients (n=22) and donors (n=14). The abdominal aorta from AAA patients exhibit extreme structural abnormalities characterised by a destructive connective tissue remodelling and an important inflammatory infiltrate, as described previously (Camacho et al., 2013; Golledge et al., 2006). HDAC1, 2, 4 and 7 mRNA levels were significantly upregulated in AAA (between 15- and 5-fold) compared with healthy controls, whereas no differences were observed for HDAC3, 5 and 8 (Fig. 1A). The upregulation of HDACs was confirmed by western blot (Fig. 1B) and was further assessed by immunohistochemistry. A strong immunostaining for HDAC1, 2, 4 and 7 was detected in the media layer of aneurysmal samples, specifically in VSMCs and in areas that were enriched with inflammatory cells (Fig. 1C). Double-immunofluorescence analysis in human AAA samples corroborate that HDAC1, 2 and 4 are expressed by both smooth muscle α-actin cells (VSMCs) and CD3-positive cells (T cells) (Fig. S1). Consistently, the enhanced expression of HDACs in AAA was associated with a decrease in the degree of histone H3 acetylation (Fig. 1B,D). Clinical data of patients and donors are shown in Table 1.
The Ang-II-infused ApoE−/− mouse is a well-established model of AAA (Daugherty et al., 2000). Similarly to what is observed in humans, the expression of HDAC1, 2, 4 and 7, and also that of HDAC3, was increased in the aorta of ApoE−/− mice after 4 weeks of Ang-II infusion, compared with saline-infused animals, whereas mRNA levels for HDAC5 and 8 remained unchanged (Fig. S2).
HDAC inhibitors reduced mortality rates in Ang-II-infused ApoE−/− mice and limited the incidence and severity of AAA
Owing to the strong upregulation of HDACs in AAA, we aimed to determine whether HDAC inhibition could limit aneurysm development. Ang-II-infused ApoE−/− mice were treated with two well-known class-selective HDAC inhibitors, MS-275 (an inhibitor of class I HDACs) and MC-1568 (a class IIa HDAC inhibitor) (Beckers et al., 2007; Fleming et al., 2014). Ang-II infusion for 4 weeks increased systolic blood pressure and mean arterial pressure, but neither affected lipid profile nor body weight, as previously described (Daugherty et al., 2000). None of these parameters was altered by the inhibition of HDAC activity (Table S1). Furthermore, both inhibitors markedly reduced the cardiac hypertrophic response to Ang II (data not shown), in agreement with previous data (Kee et al., 2006; Kong et al., 2006).
Ang-II infusion resulted in the early death of some animals owing to aortic rupture, whereas HDAC inhibition delayed death episodes (Fig. 2A). The percentage of Ang-II-infused ApoE−/− mice that survived over the course of the study was 62%, with deaths occurring after the fifth day post-infusion. MS-275 treatment did not significantly improve the survival rate, although this drug delayed fatal events by 1 week. Interestingly, all Ang-II-infused ApoE−/− mice treated with MC-1568 survived until the end of the study (Fig. 2A). Ang-II-infused ApoE−/− mice treated with doxycycline, a broad-spectrum MMP inhibitor that inhibits AAA development in this model (Manning et al., 2003), showed a 70% survival rate.
In order to follow-up AAA progression throughout the study, aortic diameter at 0, 14 and 28 days after Ang-II infusion was monitored and recorded by ultrasonography (Fig. 2B). Ang II progressively increased abdominal aortic diameter, whereas HDAC inhibitors significantly decreased aortic diameter 14 days after treatment and no further progression was observed (Fig. 2B). The MMP inhibitor doxycycline, used as a positive control, produced a similar effect. Data in Fig. 2C and ultrasound images at the end of the study (Fig. 2D) evidenced the significant attenuation of aortic diameter exerted by these drugs.
Ang-II infusion in ApoE−/− mice induced the formation of suprarenal aneurysm and structural alterations in the whole aorta, whereas no spontaneous AAAs were observed in saline-infused mice (Fig. 2E, left panel). After Ang-II infusion, only 23% of mice treated with MC-1568 and 54% of those receiving MS-275 developed AAA, in contrast to the high incidence in ApoE−/− mice (92%). The preventive effect of MS-275 on AAA incidence was similar to that exerted by doxycycline (50%; Fig. 2E, right panel). Control mice injected with vehicle [0.5% carboxymethylcellulose or 50% dimethyl sulfoxide (DMSO) alone] did not show alterations in aortic diameter or in aorta morphology (not shown). At the end of the procedure, Ang-II-infused mice exhibited the most severe forms of AAA (Table 2). The treatment with HDAC inhibitors reduced the severity of aneurysmal lesions. Mice from the MC-1568 group showed a low incidence of aneurysms and less severe forms of them, whereas the MS-275 group exhibited an intermediate phenotype.
Histological examination of the suprarenal aorta showed that HDAC inhibitors attenuated the Ang-II-induced disturbance in overall abdominal aorta morphology. These drugs blocked the marked increase in aortic wall thickness and collagen deposition observed in Ang-II-infused ApoE−/− mice [Fig. 3A (upper panels), B]. Additionally, orcein staining showed that inhibition of either class I or class II HDACs reduced the disruption of elastic lamina that was frequently observed in Ang-II-infused mice [Fig. 3A (lower panel), C]. Immunostaining for smooth muscle α-actin showed that VSMC density was remarkably reduced in the media of aneurysm sections from Ang-II-infused mice compared with controls. Interestingly, both HDAC inhibitors prevented such an effect (Fig. 3D). Apoptosis is a key factor that contributes to medial degeneration in AAA (Thompson et al., 1997). In agreement with the reduction in medial VSMC density observed in Ang-II-infused animals, we detected cleaved-caspase-3-positive cells in these mice, an effect that was prevented by MC-1568 or MS-275 (Fig. S3A), and similar results were obtained by TUNEL assay (Fig. S3B).
HDAC inhibitors attenuate the Ang-II-induced systemic and vascular inflammatory response
The activation of the inflammatory response is a well-established process in both human aneurysm and Ang II-induced experimental AAA (Freestone et al., 1995; Shimizu et al., 2006). In agreement, a marked increase in the accumulation of infiltrated macrophages, T-cells and neutrophils into the vessel wall of Ang-II-infused ApoE−/− mice was revealed by immunohistochemistry, as compared to animals receiving saline (Fig. 4A). Mac3-, CD3- and neutrophil-elastase-positive cells were abundant in ApoE−/− mice infused with Ang II but not in control aortas, whereas treatment with either HDAC inhibitor significantly reduced the immune infiltrate (Fig. 4B-D). Furthermore, the drastic upregulation in the expression of EMR-1 (a macrophage marker) and Elane (a neutrophil marker) induced by Ang II was attenuated by HDAC inhibition (Fig. 4E,F). Likewise, immunohistochemistry and real-time PCR analysis evidenced that both MC-1568 and MS-275 prevented the vascular increase in MCP-1 (monocyte chemotactic protein-1) expression triggered by Ang II in ApoE−/− mice (Fig. 5A,B) and, similarly, these drugs reduced the enhanced expression of other pro-inflammatory mediators, including cyclooxygenase 2 (COX-2; Fig. 5C) and interleukin (IL)-1β and -6 (Fig. 5D). Likewise, the striking increase in the circulating levels of IL-6 and IL-2 in ApoE−/− mice in response to Ang II was significantly reduced by class I and II HDAC inhibition (Fig. 5E). HDAC inhibitors also attenuated the inflammatory response triggered by Ang II in human VSMCs in culture (Fig. S4A-C). These results suggest a direct implication of HDACs in the regulation of the inflammatory response in this AAA model.
HDAC inhibitors decrease MMP expression/activity in Ang-II-infused ApoE−/− mice
We assessed the repercussion of HDAC inhibition on the proteolytic process mediated by MMPs in Ang-II-induced aneurysm. MMP-2 and MMP-9 are the main isoforms that degrade elastic fibres and collagen in mouse models of AAA (Davis et al., 1998; Pyo et al., 2000; Longo et al., 2002; Luttun et al., 2004). As expected, a marked induction of MMP-2 and MMP-9 expression was detected in the abdominal aorta from Ang-II-infused ApoE−/− mice, as evidenced by immunofluorescence (Fig. 6A) and real-time PCR analysis (Fig. 6B). This increase was significantly prevented by both MC-1568 and MS-275 (Fig. 6A,B). In accordance, zymography assays performed with abdominal aorta lysates demonstrate that Ang II induced the zymogens and active forms of MMP-2 and MMP-9 (Fig. 6C). Interestingly, class I and class IIa HDAC inhibitors elicited a significant attenuation in the activity of these MMPs and a reduction in their pro-form levels (Fig. 6C). Similarly, these drugs limited the induction of MMP-2 expression evoked by Ang II in human VSMCs in culture (Fig. S4D).
Therapeutic strategies in AAA are restricted to invasive surgical repair of those AAAs under a high risk of rupture and, unfortunately, there are still no effective pharmacological drugs that limit AAA progression and rupture. Although statins, doxycycline, vitamin E, COX-2 inhibitors, angiotensin-converting enzyme inhibitors and Ang-II-receptor blockers have been suggested to reduce AAA expansion (Golledge et al., 2006; Samson, 2012; Dodd and Spence, 2011; Manning et al., 2003), none of these therapies has conclusively demonstrated their clinical benefit. Therefore, the search for new therapeutic targets in AAA is a challenge that requires a deep understanding of the mechanisms involved in this disease.
In recent years, an increasing number of studies have reported that an exacerbated expression/activity of HDACs could be detrimental in multiple pathologies, including cardiovascular diseases (Pons et al., 2009; Trivedi et al., 2007; Findeisen et al., 2011). Despite the outstanding progress in the knowledge of the pathophysiological role of HDACs in cardiovascular pathologies, little is known about their involvement in AAA formation, expansion and rupture. Our results evidence for the first time the induction of HDACs in human AAA. Furthermore, in an experimental model of Ang-II-induced AAA, class I and class IIa HDAC inhibitors largely reduced AAA progression, suggesting that HDACs could play an important role in aneurysm formation.
In human aneurysmal samples, specific HDACs from class I and II were induced, in particular HDAC1, 2 (both class I), 4 and 7 (both class IIa), supporting their involvement in the human pathology. This induction was observed in VSMCs and mainly in the inflammatory infiltrate. In accordance, we observed a reduction in the degree of histone H3 acetylation in human AAA most likely secondary to the induction of HDAC expression. In contrast, an increased histone H3 acetylation has been reported in VSMCs in culture from patients with Marfan and non-Marfan thoracic aneurysm (TAA) related to a slight enhancement in histone acetyltransferase activity (Gomez et al., 2011). Hence, these divergent epigenetic mechanisms that seem to participate in AAA and TAA could explain the different etiopathogenesis of these diseases.
Interestingly, a similar expression pattern of class I and class II HDACs was found in the abdominal aorta of ApoE−/− mice infused with Ang II and, hence, the HDAC-dependent changes detected in this model resemble those observed in human disease. Owing to the reversibility of histone acetylation, HDAC inhibition raised as an attractive therapeutic option. Our results demonstrate that systemic administration of selective class I and class IIa HDAC inhibitors in this model delayed mortality and limited AAA expansion and severity. An early study reported that Metacept, a highly cytotoxic class I HDAC inhibitor (Shehu-Xhilaga et al., 2009), ameliorated Ang-II-induced aneurysm incidence without any beneficial effect on mortality rates (Vinh et al., 2008). However, we observed that MS-275, a class I HDAC inhibitor in ongoing clinical trials for oncological diseases, retards death episodes and, more interestingly, under our experimental approach, MC-1568 completely prevented mortality and reduced AAA severity. These results suggest that class IIa HDAC inhibitors constitute a promising therapeutic option for AAA patients. The beneficial effect of HDAC inhibitors was not related to either an improvement in the lipid profile or a reduction in Ang-II-induced high blood pressure. In fact, previous data showed that AAA formation in Ang-II-infused ApoE−/− mice is independent of the increase in arterial pressure (Cassis et al., 2009; Lu et al., 2012).
VSMC apoptosis is another key feature of human AAA that contributes to the degeneration of the vascular wall (Thompson et al., 1997). Similarly, α-actin immunostaining evidenced a reduction in VSMC density in the abdominal aorta of Ang-II-infused ApoE−/− mice that was associated with higher levels of both active caspase 3 (cleaved caspase 3), frequently used as a marker of apoptosis (Yamanouchi et al., 2012), and TUNEL staining. HDAC inhibitors attenuated these effects, suggesting that, in the vascular wall, these drugs elicit anti-apoptotic responses that could favour arterial wall stabilisation.
HDAC inhibitors also attenuated the extracellular matrix (ECM) disorganisation and degradation, which plays a pivotal role in the progressive aortic expansion observed in Ang-II-infused ApoE−/− mice. The MMP-mediated proteolytic activity in the vessel wall is critically involved in ECM breakdown and AAA formation. MMP-2 and MMP-9 have been the most extensively studied MMPs in the context of this disease and show the strongest elastinolytic activity (Davis et al., 1998; Pyo et al., 2000; Longo et al., 2002; Luttun et al., 2004). In accordance with the lower elastin degradation and reduced collagen disorganisation exhibited by MC-1568- or MS-275-treated animals, these drugs elicited a significant reduction in vascular MMP-2 and MMP-9 expression and activity compared with Ang-II-infused ApoE−/− mice. Indeed, epigenetic mechanisms dependent on HDAC activity are involved in the control of ECM synthesis and deposition (Pons et al., 2009), and modulate MMP expression and activity (Liu et al., 2003; Culley et al., 2013).
Humans with AAA exhibit an increase in inflammatory mediators in the vascular wall and enhanced levels of circulating cytokines and chemokines (Freestone et al., 1995; Shimizu et al., 2006). In agreement, Ang II induced a marked infiltration of macrophages, lymphocytes and neutrophils in the arterial wall from ApoE−/− mice. Interestingly, HDAC inhibitors markedly decreased the inflammatory infiltrate in the aortic wall, accompanied by a significant reduction in the vascular expression of inflammatory markers, including COX-2, IL-1β, MCP-1 and IL-6, and an amelioration of systemic inflammation. Previous studies reported that attenuation of inflammation in AAA animal models inhibits aortic dilation, whereas an aggravated inflammatory response is associated with the progression and rupture of large aneurysms (Freestone et al., 1995; Saraff et al., 2003; Rateri et al., 2011). Interestingly, HDAC inhibitors exhibit anti-inflammatory and immunomodulatory functions regulating macrophage and lymphocyte responses under inflammatory conditions (Leoni et al., 2002; McKinsey, 2011; Licciardi et al., 2013). Based on the inhibitory effect of MC-1568 and MS-275 on local and systemic inflammation, our data suggest that HDACs could contribute to vascular inflammation in AAA through the control of the function of inflammatory cells and the consequent increase in cytokine and chemokine production.
In summary, we have shown a strong upregulation of specific HDACs in human AAA and that HDAC inhibitors limit aneurysm progression in a mouse model. Improvements in ECM remodelling, vascular apoptosis and inflammation could account for the global benefit of these drugs. Furthermore, in our preclinical model, some treatment regimens could be particularly efficient in improving survival, suggesting that these drugs might constitute a new pharmacological strategy for the treatment and stabilisation of AAA. Further research is warranted to define the molecular targets involved in their vast benefit.
Human samples
Human aneurysmal samples were obtained from patients undergoing open repair for AAA at the Hospital de la Santa Creu i Sant Pau (HSCSP; Barcelona, Spain); healthy aortas were obtained from multi-organ donors as previously described (Camacho et al., 2013). Approval to use the discarded human tissue was given by the Ethics Committee of the HSCSP. Research has been carried out in accordance with the Declaration of Helsinki. Participation in the study of patients and control subjects was based upon informed consent of patients or legal representatives.
Abdominal aorta segments were obtained from all patients (n=22) and control subjects (n=14), following strict standard operating procedures and ethical guidelines. Samples of control subjects had no post-mortem evidence of abdominal aorta aneurysm, atherosclerotic plaques or other medical conditions that affect the study. Samples were rapidly collected and stored at −80°C for subsequent RNA and protein studies or processed for immunohistochemical analysis.
Cell culture
Human aortic VSMCs were obtained from non-atherosclerotic arteries of hearts removed in transplant surgeries at the HSCSP by using a modification of the explant technique. Human VSMCs were cultured in M199 (Gibco, Carlsbad, CA) supplemented with 20% fetal calf serum (FCS), 2% human serum, 2 mmol/l L-glutamine, 100 U/ml penicillin G and 0.1 mg/ml streptomycin (Camacho et al., 2013; Orriols et al., 2014). VSMCs from at least four donors were used. Studies were performed with cells in passages 3 to 6. The research was performed in accordance with the Declaration of Helsinki and approved by the HSCSP Ethics Committee. Purity of VSMCs was confirmed by positive immunohistological staining for α-smooth muscle actin and by the negative staining for von Willebrand factor. Mycoplasma contamination was periodically analysed and excluded. For experimental procedures, subconfluent cells were arrested in medium containing 0.4% FCS for 24 h and stimulated with Ang II (10−7 M) for 24 h. Alternatively, VSMCs were pre-treated (1 h) with MC-1568 (5 μM; Selleck Chemicals) or MS-275 (2 μM; Selleck Chemicals). Treatments did not induce cytotoxicity as determined by the MTT assay (Roche Diagnostics, Indianapolis, IN).
Animal handling
Animals were housed in the Animal Experimentation Unit of the Cardiovascular Research Center (CSIC-ICCC, Barcelona, Spain) in a controlled, specific pathogen-free environment under standard light-dark cycle (12 h light/dark cycle) and temperature (21±1°C) conditions, and were fed ad libitum with a standard commercial diet (Harlan Iberica SL, Barcelona, Spain). All animal handling procedures were performed in compliance with the principles and guidelines established by the Spanish Policy for Animal Protection RD53/2013, which meets the European Union Directive 2010/63/UE on the protection of animals used for experimental and other scientific purposes, and all procedures were reviewed and approved by the Ethical Committee at the Centro de Investigación Cardiovascular as stated in Law 5/1995, 21 June, passed by the Generalitat de Catalunya.
The apolipoprotein-E-deficient (ApoE−/−) mouse infused with angiotensin II (Ang II) was used as a model of AAA as previously described (Daugherty et al., 2000). 10-week-old male ApoE−/− mice (ApoE−/−; B6.129P2-Apoetm1Unc/J) were obtained from Charles River UK Ltd (Kent, UK). Mice were acclimated 1 week prior to the study. Ang II (1000 ng kg−1 body weight min−1; Sigma-Aldrich, St Louis, MO) was infused via osmotic minipumps (model 1004, Alzet, DURECT Corporation, CA) implanted subcutaneously into the interscapular space of isoflurane-anaesthetised mice for 28 days. Animals were randomly distributed in five experimental groups: untreated Ang-II-infused mice (n=20); two groups of Ang-II-infused mice that received an intraperitoneal injection of histone deacetylases (HDACs) inhibitors (Selleck Chemicals, Houston, TX) [MC-1568 (a class-IIa-HDAC-specific inhibitor; 50 mg kg−1 body weight) or MS-275 (a class-I-HDAC-specific inhibitor; 20 mg kg−1 body weight) (n=13 each group)] 3 days a week; a group of Ang-II-infused ApoE−/− mice receiving the broad-spectrum metalloproteinase (MMP) inhibitor doxycycline (doxycycline hyclate, Sigma-Aldrich; 30 mg kg−1 body weight day−1), which has previously been demonstrated to inhibit AAA formation in this model (Manning et al., 2003) (administered in the drinking water; n=6); and a control group of ApoE−/− mice infused with saline (n=18). Doses of HDAC inhibitors were chosen according to previous in vivo studies (Nebbioso et al., 2009; Spallotta et al., 2013; Dalgard et al., 2008), and treatments started 3 days before minipump implantation.
For the implantation of osmotic minipumps, mice were anaesthetised with isofluorane (2%), which has a rapid effect on animals. Anaesthetic depth was confirmed by loss of blink reflex and/or lack of response to tail pinch. The procedure takes about 15 min/mouse. Recovery after surgical procedures was carried out using aseptic techniques in a dedicated approved surgical area. Antibiotics (penicillin, 450,000 u kg−1, intramuscular) and analgesics (buprenorphine 0.05 mg kg−1, subcutaneous) were given immediately after surgery to prevent infection and discomfort. The animals were kept warm in a heating pad until awake after surgery, and observed carefully by the investigators throughout the post-surgery period. At the end of the experimental procedures, mice were euthanised via isofluorane overdose and the aortas were immediately harvested, examined for the presence of an AAA and appropriately processed for further studies.
Non-invasive measurement of systolic blood pressure
Systolic blood pressure (SBP) and mean arterial pressure (MAP) were non-invasively measured in conscious mice prior to and following treatment using the tail-cuff plethysmography method (Panlab, Harvard Apparatus). Mice were trained for tail-cuff measurements over a period of 1 week. Blood pressure measurements were performed at the same time (between 9 a.m. and 11 a.m.) in order to avoid the influence of the circadian cycle. Mean blood pressure values were taken from ten consecutive measurements (Orriols et al., 2014).
Basic measurements of ultrasound recording for abdominal aortas
Mice were anaesthetised with 1.5% isofluorane inhalation and were lightly secured in the supine position to a warming platform. After shaving the precordium, an abdominal echography was performed using a Vevo 2100 ultrasound with a 30 MHz transducer applied to the abdominal wall to record abdominal aorta (VisualSonics, Toronto, Canada). Abdominal aortas with external diameters ≥1.5 mm were considered as an aneurysm. All primary measurements were made from images captured on cine loops of 100 frames at the time of the study using the software provided by the echography machine.
The severity of the aneurysm was based on a 4-point grading scale previously described in detail (Manning et al., 2003): type 0, no aneurysm; type I, dilated lumen in the suprarenal region of the aorta with no thrombus; type II, remodelled tissue in the suprarenal region that frequently contained thrombus; type III, a pronounced bulbous form of type II that contained thrombus, and type IV, a form in which there are multiple AAAs containing thrombus.
Total mRNA and protein isolation from tissues
The RNeasy Mini Kit (Qiagen, Venlo, Netherlands) was used to isolate total RNA from human AAA samples following the manufacturer's recommendation. Protein lysates from human AAA were prepared in a RIPA buffer [150 mM NaCl, 1% (v/v) Triton X-100, 0.5% (w/v) sodium deoxycholate, 0.1% (w/v) SDS, 2 mM EDTA, 50 mM Tris-HCl pH 8] by using a tissue homogenizer and following a standard protocol.
In the case of animal studies, after removal of adventitial fat from mouse aortas, the aneurysm was separated, snap-frozen in liquid nitrogen and stored at −80°C. Total RNA and protein isolation from mouse abdominal aorta was performed using the Tripure reagent (Roche Diagnostics, Indianapolis, IN) following the manufacturer's instructions. RNA integrity was determined by electrophoresis in agarose gels and was quantified by a NanoDrop 1000 Spectrophotometer (Thermo Scientific).
Real-time PCR
DNase-I-treated total RNA (1 μg) was reverse transcribed into cDNA using the High Capacity cDNA Archive Kit (Applied Biosystems, Foster City, CA) with random hexamers. Quantification of mRNA levels was performed by real-time PCR using an ABI PRISM 7900HT sequence detection system (Applied Biosystems) and specific primers and probes provided by the Assay-on-Demand system (Applied Biosystems) for human HDAC1 (Hs02621185_s1), HDAC2 (Hs00231032_m1), HDAC3 (Hs00187320_m1), HDAC8 (Hs00218503_m1), HDAC4 (Hs01041638_m1), HDAC5 (Hs00608366_m1), HDAC7 (Hs00248789_m1), MMP2 (Hs00234422_m1), interleukin 1β (IL1β; Hs00174097_m1), chemokine (C-C motif) ligand 2 (CCL-2 or MCP-1; Hs00234140_m1) and prostaglandin-endoperoxide synthase 2 (PTGS2 or COX-2; Hs00153133_m1). 18S rRNA (4319413E) was used as an endogenous control for human samples. To analyse mRNA levels in mouse tissues, we used TaqMan fluorescent real-time PCR primers and probes, provided by Applied Biosystems or Integrated DNA technologies as follows: IL6 (Mm00446191_m1), IL-1β (Mm00434228_m1), MCP-1 (Mm00441242_m1), COX-2 (Mm00478374_m1), HDAC1 (MmPT.58.14183463), HDAC2 (MmPT.58.13367660), HDAC3 (MmPT.58.13259320), HDAC4 (MmPT.58.17110912), HDAC5 (MmPT.58.16075245), HDAC7 (MmPT.58.13463976), HDAC8 (Mm01224980_m1), MMP-9 (MmPT.58.10100097), MMP-2 (MmPT.58.9606100), EMR-1 (MmPT.56a.11087779) and Elane (elastase, neutrophil expressed; Mm00469310_m1). As endogenous controls for mouse samples, glyceraldehyde 3-phosphate dehydrogenase (GAPDH; Mm99999915_g1) and 18S rRNA (4319413E) were used. Quantitative RT-PCR was carried out in an ABI PRISM 7900HT Sequence Detection System (Applied Biosystems) using the following conditions: 2 min at 50°C, 10 min at 95°C followed by 40 cycles of 15 s at 95°C and 1 min at 60°C. Relative mRNA levels were determined using the 2−ΔΔCt method.
Western blot
Tissue lysates were separated by SDS-PAGE and transferred to 0.45 μm polyvinylidene difluoride membranes (Immobilon, Millipore, Merck KGaA, Darmstadt, Germany). Blots were incubated with antibodies directed against HDAC1 (NBP1-67590), HDAC2 (NB100-91802) and HDAC4 (NBP1-67592) purchased from Novus Biologicals (R&D Systems Europe Ltd, Abingdon, UK); antibodies against HDAC7 (ab166911) and histone H3 acetyl K18 (Ac-H3K18; ab1191) were obtained from Abcam (Cambridge, UK). All were used at 1:1000 dilution. Equal loading of protein in each lane was verified by β-actin (A5441, Sigma-Aldrich).
Immunohistochemistry, immunofluorescence and histology
Aortic samples were fixed in 4% paraformaldehyde/0.1 M phosphate buffered saline (PBS; pH 7.4) for 24 h and embedded in paraffin. Aortic sections from human and mouse samples (5 μm) were deparaffinised in xylene, rehydrated in graded ethanol, and treated with 0.3% hydrogen peroxide for 30 min to block peroxidase activity. Then, samples were blocked with 10% of normal serum and incubated with antibodies against LAMP-2/Mac-3 (sc-19991, 1:250, Santa Cruz Biotechnology Inc., Europe), CD3 (A0452, 1:100, Dako, Agilent Technologies Co., Hamburg, Germany), MCP-1 (sc-1785, 1:100, Santa Cruz Biotechnology Inc.), Elane (M0752, Dako, Agilent Technologies Co., Hamburg, Germany), cleaved-caspase 3 (9661, 1:100, Cell Signaling, Boston, MA), HDACs and Ac-H3K18 overnight at 4°C (both at 1:100). After washing, samples were incubated for 1 h with a biotinylated secondary antibody (Vector Laboratories, Peterborough, UK). After rinsing three times in PBS, standard Vectastain avidin-biotin peroxidase complex (ABC; Vector Laboratories) was applied, and the slides were incubated for 30 min. Colour was developed using 3,3′-diaminobenzidine (DAB) and sections were counterstained with haematoxylin before dehydration, clearing and mounting. Negative controls, in which the primary antibody was omitted, were included to test for non-specific binding.
For immunofluorescence studies, mouse abdominal aorta sections were blocked in PBS containing 5% BSA and 0.1% Triton X-100, and then sections were incubated with a polyclonal antibody against MMP-9 (sc-10737, 1:100, Santa Cruz Biotechnology), a rabbit monoclonal antibody against MMP-2 (ab51125, 1:100, Abcam), or a rabbit polyclonal antibody against α-actin (ab5694, Abcam), α-actin (M0851, 1:200, Dako, Agilent Technologies Co., Hamburg, Germany), CD3 and HDACs overnight at 4°C. After washing, rings were incubated with an anti-rabbit IgG conjugated to TRITC or FITC probes (Dako) for 1 h at room temperature (RT). After washing, slides were mounted with SlowFade Gold antifade reagent with DAPI (Molecular Probes, Life Technologies, Madrid, Spain) and immunofluorescent signals were viewed using a Nikon Eclipse 55i fluorescence microscope. Results were quantified and expressed as percentage of positive area versus total area and as positive cell number in independent sections of AAA for CD3, Mac-3, MCP-1, neutrophil elastase and cleaved-caspase 3. Negative controls, in which the primary antibody was omitted, were included to test for non-specific binding. The histological characterisation of aortic samples was performed by Masson's trichrome staining. Furthermore, to visualise elastic fibre integrity, arterial sections were stained with orcein using a commercial kit (Casa Álvarez, Madrid, Spain).
Double-fluorescence immunostaining
For antigen colocalization studies, double-fluorescence immunostaining was performed using a sequential method. After deparaffinization, antigen retrieval and permeabilization with PBS-0.1% Triton X-100, sections were blocked with PBS containing 5% albumin (4°C overnight). Incubation with rabbit polyclonal antibodies against HDAC1, HDAC2 and HDAC4, and those against smooth muscle α-actin or CD3 (1:100), were then sequentially applied. After washing the secondary antibodies, donkey anti-mouse IgG conjugated to Alexa Fluor 542, and chicken anti-rabbit IgG conjugated to Alexa Fluor 488 (Molecular Probes, Life Technologies) were applied for 1 h at RT. Negative controls, in which the primary antibody was omitted, were also included. Samples were then mounted with ProLong Gold antifade reagent with DAPI (Molecular Probes, Life Technologies Co., Eugene, OR). Images were obtained using an SP5 Leica confocal microscope.
Gelatin zymography
The relative enzymatic activities of MMP-9 and MMP-2 in murine abdominal aorta lysates were measured by zymography. Protein lysates were prepared in RIPA buffer supplemented with complete protease inhibitor cocktail (Roche). Proteins (20 μg) were resolved by 10% SDS-polyacrylamide gels copolymerised with 1 mg ml−1 of porcine skin type A gelatin (Sigma-Aldrich) as a substrate for MMP enzymatic activity, and run at 4°C for 4-6 h. After electrophoresis, the gels were rinsed twice for 30 min at RT in 2.5% Triton X-100 (Sigma), and then incubated in substrate buffer [50 mM Tris-HCl, 10 mM CaCl2 (Merck), 0.02% (w/v) N3Na (Fluka), pH 8] for 18-20 h at 37°C. Gels were dyed with one tablet of PhastGel™ Blue R (GE Healthcare) in 10% acetic acid. Areas of gelatinolytic activity appeared as clear bands on a blue background where the protease has digested the substrate. Gels were finally scanned with a GS-800 calibrated imaging densitometer (Bio-Rad) and quantitative densitometric analysis of digested bands was performed using Quantity-One software (Bio-Rad) (Rodríguez-Calvo et al., 2015).
Enzyme-linked immunosorbent assay (ELISA)
IL-6 and IL-2 concentrations in plasma were determined using the Quansys Q-plex mouse cytokine system (Quansys Biosciences, UT).
Determination of the lipid profile in plasma
Lipids were quantified with a colorimetric assay in plasma from all groups of mice by using specific reagents for triglycerides (Gernon, GN90125; RAL Técnica para el Laboratorio S.A., Barcelona, Spain), total cholesterol (Gernon, GN20125) and high-density lipoprotein (HDL) cholesterol (Gernon, GN20525), and a multicuvette rack reader (Clima MC-15, RAL Técnica para el Laboratorio S.A., Barcelona, Spain) following the manufacturers' instructions. HDL was quantified by the phosphotungstic acid method using a COBAS 6000 analyzer (Roche Diagnostics, USA) as described (Ribas et al., 2004).
TUNEL assay
Recombinant terminal deoxynucleotidyl transferase (rTdT)-mediated nick-end labelling (TUNEL) was performed using the Dead End Fluorometric Tunel System (Promega) according to the manufacturer's guidelines. Four-micrometer paraffin sections were deparaffinised, fixed in methanol-free paraformaldehyde before and after proteinase K treatment at 20 μg/ml for 8-10 min at RT. The sections were incubated with the nucleotide mixture (which included fluorescein-tagged dUTP) and rTdT enzyme for 1 h at 37°C. The slides were mounted using SlowFade Gold antifade reagent with DAPI (Molecular Probes, Invitrogen) and immunofluorescent signals were viewed using a fluorescence microscope (Nikon Eclipse 55i).
Statistical analysis
Data were expressed as mean±s.e.m. and were analysed using the GraphPad Prism 4.0 software (GraphPad, USA). Human and animal population data were expressed as mean±s.e.m. Values of P≤0.05 were considered significant. Differences between specified groups were analysed using the Student's t-test (two-tailed) comparing two groups. One-way ANOVA and the Bonferroni test were used to determine the statistical significance of the observed differences between the studied groups. When normality failed, we used the Mann–Whitney rank sum test to compare two groups and Kruskal–Wallis one-way ANOVA on ranks for multiple comparisons (Dunn's method). Differences in the trends for mortality between groups were determined using Kaplan–Meier test and differences in the percentage of incidence of AAA were analysed by Chi square test (χ2).
Authors are grateful to Montserrat Gómez and Iris Rodríguez for their technical support, and Dr J. Julve (Institut de Recerca Hospital de la Santa Creu i Sant Pau, Barcelona) for his recommendations and assistance to determine plasma HDL levels.
Competing interests
The authors declare no competing or financial interests.
Author contributions
C.R. designed and supervised the study, interpreted data and wrote the manuscript. M.G. performed experiments and analysed and interpreted data; additionally, M.G. wrote a first draft of the manuscript. S.V. performed experiments, analysed and interpreted data. J.D. and M.C. were responsible for the clinical aspects of the study, participated in data collection and analysis. M.O., J.A.R., J.M.-G. conceived specific experiments and analysed data. S.A. participated in in vivo studies. All authors were involved in writing the paper and gave their final approval of the submitted version.
This work was supported by the Spanish Ministerio de Economía y Competitividad (MINECO)-Instituto de Salud Carlos III (ISCIII) [grants PI15/01016 and PI12/01952 to C.R.; SAF2012-40127 to J.M.-G.; SAF2013-46707-R to M.C., RD12/0042/0053 to J.M.-G. and C.R.; RD12/0042/0051 to MC and RD12/0042/0009 to J.A.R.]. The study was co-founded by Fondo Europeo de Desarrollo Regional (FEDER)-The way to build Europe. M.G. was supported by funds provided by ISCIII (Sara Borrell program).
Supplementary information
Supplementary information available online at http://dmm.biologists.org/lookup/suppl/doi:10.1242/dmm.024513/-/DC1
- Received December 21, 2015.
- Accepted March 14, 2016.
- © 2016. Published by The Company of Biologists Ltd
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.


text:
Tires vs. Tracks
John Deere or Case? Ford or Chevy? Tire or tracks? After decades of discussion, farmers still disagree over the appropriate answers to these questions. While we won't weigh in on Deere vs. Case or Ford vs. Chevrolet, we do have an opinion (although it might be a bit biased) about the superiority of tires to tracks and we're pretty sure that, after reading this, you'll agree that tires are equal to or exceed tracks in performance, while costing substantially less.


text:
01543 363739
Meet The Team
Rob Morris - Senior Web DeveloperHis key roles are producing and maintaining every element of web development within the company. Rob's skill base is drawn on to produce new sites, further develop existing sites, invest time and practice all new technology that will ensure our client base are kept at the very edge of advancement.
Further skills include Search Engine Optimisation, e-commerce, graphic design.


text:
Happy Monday everyone! How was your weekend? Mine was filled with the usual soccer games, football, and birthday parties. Sadly, there wasn't too much creative time:) But, today is the last day of September! October is always full of creative opportunities with more birthdays, anniversaries, and Halloween. I'm already planning a trip to the pumpkin patch and a corn maze. Until then, let's look at these cards...
Jillibean Soup
Patterned Paper: Grandma's Lima Bean Soup/Lean Ham
Pea Pod Parts: Coconut Lime Soup
Other: Bazzill Basic kraft cardstock.
Let's Be Adventurers Card by Nancy Damiano
Jillibean Soup
Patterned Paper: Happy Camper Stew/Scores of Smores, Bite Size Bits; Irish Farm House Soup/Barley
Coordinating Shape Stickers: Happy Camper Stew
Pea Pod Parts: Happy Camper Stew
Sugar Picks: Sunshine
Thinking Of You Card by Emily Pitts
Jillibean Soup
Patterned Paper: Christmas Cheer Chowder/Folded Flakes; Cold Avocado Soup/Ripe Avocado, Green Onions; Happy Camper Stew/4 Parts Great Outdoors
Pea Pod Parts: Cold Avocado Soup
Sugar Picks: Swallowtail
Wood Flags: Thinking Of You
Bean Stalks: Red Baker's Twine
Other: Therm O Web adhesive, Scotch adhesive, Glue Arts, Bazzill, Gartner.
Class Of Your Own Card by Laurel Seabrook
Jillibean Soup
Patterned Paper: Happy Camper Stew/Scores of Smores, Dash of Danger, 4 Parts Great Outdoors, Bite Size Bits
Pea Pod Parts: Happy Camper Stew
Coordinating Label Stickers: Happy Camper Stew
Corrugated Shapes: People


text: The Gummibär Easter Egg Giveaway has begun! THREE lucky winners will each receive an awesome Gummibär Easter egg prize completely at random. The combined value of these eggs is over $100, with the biggest egg being worth over $70! Here's what's exactly in each Easter egg: Green Egg: 12" plush toy Gummy Candy Sour Gummy Candy Reusable […]Read More


text:
What are Cookies?
'Cookies' are small text files that are stored by your browser (for example, Internet Explorer or Safari) on your computer or phone. Cookies allow websites and blogs to store things like user preferences. They serve as a 'memory' of sorts for the website so that it can recognize you when you come back and respond appropriately.
How are Cookies used on TheThingsWellMake.com?
A visit to this blog can generate the following types of cookies:
- Anonymous analytical cookies – These cookies gather general statistics about things like the number of visitors to the blog, but they can't be used to identify individuals.
- Third party advertising cookies – These cookies are used to anonymously target ads to people and to track how many people have seen a particular ad, or how many people have seen it more than once.
Examples of third party advertising cookies
DoubleClick DART Cookie
- Google, as a third party vendor, uses cookies to serve ads on thethingswellmake.com.
- Google's use of the DART cookie enables it to serve ads to users based on their visit to thethingswellmake.com and other sites on the Internet.
- Users may opt out of the use of the DART cookie by visiting the Google ad and content network privacy policy here.
Some of our advertising partners may use cookies and web beacons on our site. Our advertising partners include:
- Google Adsense
- Amazon
TheThingsWellMake.com has no access to or control over these cookies that are used by third-party advertisers.
You should consult the respective privacy policies of these third-party ad servers for more detailed information on their practices as well as for instructions about how to opt-out of certain practices. thethingswellmake.com's privacy policy does not apply to, and we cannot control the activities of, such other advertisers or web sites.
How do I disable cookies?
It is usually possible to stop your browser accepting cookies, or to stop it accepting cookies from a particular website.
Modern browsers allow you to change your cookie settings. You can usually find these settings in the 'options' or 'preferences' menu of your browser. To understand these settings, the following links may be helpful, or you can use the 'Help' option in your browser for more details.
Cookie settings in Internet Explorer
Cookie settings in Firefox
Cookie settings in Chrome
Cookie settings in Safari
For more information about third party cookies generated by advertisers and how you can turn these off, go to the Your Online Choices site.
Useful links
If you would like to find out more about cookies and their use on the Internet, you may find the following links useful:


text:
Join the Glens Red Squirrel Group for a woodland walk at Breen Forest on Glenshesk Road this Sunday 19th February 10:30, we will try and see if we can spot some of the shy reds that make Breen their home and the other wildlife that may comes along. Come along and enjoy this wild jewel in the Glens.
The paths are not suited for prams and to increase the chances of seeing Red Squirrels and other wildlife, please do not take your dog on this walk. Good footwear is a must as ground is soft.


text:
I am considering getting a PMP soon, and are looking at 4 devices:
1) 5th gen iPod
2) Cowon iAudio A2
3) Creative Zen Vision
4) Archos AV500
I'm not too worried about the price (still want value for money though).
Currently i'm swaying towards the A2, as it's got USB host interface on it (read: can run USB devices through it, i.e. portable HDD's). I am wondering how far this would go, as I don't know the OS, and most USB deivces need drivers (will happily accept corrections on this fact though).
However, still not made up my mind, and still have some time before they properly get to market. Just putting this out here for some feedback/discussion from fellow geeks...


text: Home > Wine shop > White Wine > Alkoomi Wandoo Semillon 2007 Alkoomi Wandoo Semillon 2007 Varietal Semillon Region Great Southern 'Alkoomi' means 'a place we chose' and when it comes to producing impressive Semillon, they've certainly chosen well as you'll experience in this ripe, flavoursome example that has benefitted from careful cellaring. In the glass: A very impressive and still-fresh medium gold colour. On the nose: Youthful and stylish with lemon, lime, lanolin, grilled nuts and toast. On the palate: A very ripe and flavoursome wine in excellent condition showing great citrussy flavours, a creamy textural character and a fine line of balancing acidity. Light Full Bodied Dry Sweet Oak Treatment Order codeALKOO20207 Wine varietySemillon RegionGreat Southern WinemakerAndrew Cherry Food Match DescriptionConfit ocean trout, lemon oil Best DrinkingNow Alcohol Percent12.50% Read moreRead less $29.75 In any 12 $31.50 In any 6 $35.00 Per bottle Quantity (bottle) Out of Stock Availability Get free shipping for a limited time* Shop with confidence. 100% money back guarantee. 500+ Australian wineries. 40yrs experience & passion. Reviews (0) CUSTOMER REVIEWS Sort results by Highest ratings Lowest ratings Latest ratings Oldest ratings To leave a review, please Login OR Sign up Receive wines direct to your door with our Cellar Door Release plan Learn more Receive wines direct to your door with our Cellar Door Release plan Learn more You might also like Tyrrell's Vat 1 - Museum Twin Pack Price $280.00 Save $20.00 Qty | case Add to cart Meerea Park Alexander Munro Semillon 2014 $42.50 in any 12 $45.00 in any 6 $50.00 each Price | options $42.50 in any 12 bottles Qty Add to cart De Iuliis Aged Release Semillon 2013 $38.25 in any 12 $40.50 in any 6 $45.00 each Price | options $38.25 in any 12 bottles Qty Add to cart Two Blues Sauvignon Blanc 2014 1 case has been added to your cart. Cart total: xxx 1 case, 12 bottles, 3 accessories Checkout Continue Shopping


text:
Her hubby has been hard at work around there... so much has changed since I was there last time. The old boat house was torn down and a new one built in its place. A new road-sized path has been carved into the hill, ready to be paved, so we can drive all the way down to the shore. Right now we have to park at the lake house and walk down the steep hill about 100 vertical feet to get to the shore. It's not a problem going down, but going back up will knock the wind right out of you if you're not used to doing it every day.
I also finally got to see the tennis court they have up the hill from the house. I'm not sure whose property that's officially on, but all 3 houses are owned within the family. Anyway, it's a really nice court with a cute little combination storage house and viewing pavillion. All the kids were up there swinging wildly at the ball. I happened to have my own racket in the back of my car, so I went down to get it and came back up to get in on the action a little. For a while, Kate and I played back-up for Elizabeth and Spencer, hitting the ball whenever they missed it. Then we played a real game without the kids, and I won. :)
I stayed there for the night and on Saturday we went swimming. They have water trampoline anchored a short swim off their shore. The water was frigid, but as always, not so bad once you just take the plunge and deal with it. So we swam out to the trampoline. It's a big square tubular inflatable raft with a trampoline frame built into the center. There is another tube attached to one side that sticks out, and I guess that's for playing one of those games where you try to knock your opponent off the log. But it's so unstable that you can't really even stand on it in the first place. So we tried to see who could get a running start and get the most steps towards the end before falling off. That was much easier once I tightened the straps that kept it from spinning so much. There's also a slide that goes off another corner of the raft, but I didn't try that. It looks quite shallow and boring. If I'm gonna splash into that cold water, I want it to be worthwhile, like what Andy did. He got jumping high on the trampoline, then worked towards the edge and got one final hop out towards the water and did a flip. So I gave that a try too. That's fun. :) Sometimes it's enough of a challenge just to stay standing on the raft while someone else is jumping, because it moves around so much, courtesy of Isaac Newton's laws of motion. It was also quite difficult to get ONTO the thing from the water. There's a ladder up one side, but there's nothing to grab onto on the top to pull yourself up. Maybe it's easier for the kids, but Andy and I had to find something to grab, which ended up being the metal frame of the trampoline underneath the protective pad, which I'm sure the designers didn't have in mind. Oh well.
It was a constant workout to be out there. Swimming to the trampoline makes you too exhausted to climb up the ladder. Climbing up the ladder makes you to exhausted to jump up and down. Jumping up and down makes you to exhausted to swim. Then again, I'm out of shape. :)
When we had enough, I "carried" Spencer back to shore. The kids were all wearing life jackets but the adults weren't. So while I'm trying to swim back under my own power, Spencer's holding on to me, floating along effortlessly thanks to his life jacket. Actually for much of the way, he wasn't even touching me. I guess he just wanted a guide back to shore more than anything else, so he doesn't drift out into the lake. Not that I would've been in any shape to rescue him if he did. I just barely made it back to the shore as it was. (Keep in mind I was exhausted from jumping all over the trampoline. Swimming TO it was no problem.)
So that was enough swimming for me. I got showered and got lunch and everyone left soon after that, so I did too. I followed my parents back home so I could practice on their piano and bum a meal. ;) This new Jon Schmidt song I'm working on uses too much of the keyboard to practice effectively on my 61-key synth.
Well, the family's getting together again over at my parents' house today, so I'm gonna head over there sometime soon.


text:
Former Olympic champion Allen Johnson is the latest big-name athlete signed up for Gateshead's showpiece Norwich Union Super League meeting on Sunday.
Johnson, by far the fastest man over the 110m hurdles this year, faces a world-class field which includes his recent rivals from the Golden League - and rated 2 and 3 in the world rankings - Stanislav Olijars of Latvia and American Duane Ross.
Missing, though, will be the recently retired Colin Jackson. The Welshman was Johnson's biggest rival over the past decade and the current No1 believes his departure has left a huge void.
"I've been involved in some classic races with Colin over the years - and it'll be very interesting to see what the his replacement Andrew Turner can do," said the 32-year-old American.
Turner, from Nottingham, made his Great Britain team debut at European Cup in Florence.
Meanwhile, Katharine Merry is ready to make up for lost time when she lines up at Gateshead on Sunday.
For Merry, the 400m bronze medallist at the 2000 Olympics, will making her first individual appearance in two years.
A heel injury sustained in the summer of 2001 led to her being out of action for two years after surgery.
"I can't wait to get back into action on Sunday," she said. "I've been away from serious competition for much too long.
"I'm sure the nerves will be jangling. However, I've always believed in myself."
Tickets are still available via www.ukathletics.net, by phoning 0870 444 4440, or in person from the stadium.


text:
Association Matters At CAO we work to help you become more successful in your practice. From educating the public, government and insurance companies, to developing research and practice guidelines, we are here to serve you.
CAO student membership is free and students receive a wide array of benefits. CAO believes in supporting our future practitioners and has created a variety of tools to help them on their way.
Member Benefits
Thinking about a Future Career in optometry in Canada? Click here!


text:
|Recipe makes 8 servings
You will need -
* 1/2 cup carob or cocoa powder
* 5 Tablespoons Arrowroot Starch Flour
* 5 Tablespoons Tapioca Flour
* 2 cups Coconut Milk (canned)
* 2 cups unsweetened almond milk (or other nut milk)
* 1 cup agave, pure maple syrup, or honey (or replace with stevia*)
* 1 teaspoons vanilla extract (optional)
* 1 teaspoon almond extract (optional)
*Please note, if replacing liquid sweetener with Stevia powder, you will need to add additional milk (3/4 cup milk for 1 cup of liquid sweetener replaced) to avoid making pudding too thick.
|1) Stir together the cocoa or carob powder with the starches in saucepan.
|2) Add 1 cup of the coconut milk and stir until it forms a smooth paste.
|3) Stir in the rest of the milk(s) and sweetener(s).
4) Cook over medium heat, stirring frequently, until the mixture thickens and comes to a boil.
5) Remove heat and stir in the extracts.
|6) For an added touch, toast unsweetened coconut and almond slivers under broiler.
|7) Divide into 8 serving dishes and garnish with toasted almonds and coconut. Serve warm or chilled. Delicious!


text:
March, 2010
Okay, so many of you may be asking, "What the hell does that mean?" A good question…And to be honest, I had never even heard of the concept a few years ago. But, I'm a student of Caroline Myss and Robert Ohotto's work, and even studied with them in workshop intensive settings where they drove home the concept of Spiritual Congruency. Fascinating stuff.
Now back to the question: What in the hell does it mean?
Since hearing of this concept, I've found myself often mulling over what they've defined this phrase to mean… It resonates for me as a force that has brought authentic shifts to the way I approach my life. So here is the definition (as I've come to understand it):
Spiritual congruency is an alignment between your emotional self and your intellectual self.
We need both in order to be whole–and yet we often live in a fragmented state of either living through our heart (often in an overly emotionally responsive manner) or living through our heads (often in an overly intellectual manner). In the latter case, we logically calculate the path our lives take in a way that discounts the organic, fluid space the voice of our soul speaks from.
Caroline Myss brilliantly instructed me (as part of her "maiden voyage" regarding her new teaching paradigm in a workshop called "Healing and the Mystery of Grace" at the Omega center in the fall of 2007) about the energetic causes of addiction that result from the chasm between the heart and mind. She spoke in terms of our energetic anatomy system through the chakras, but she essentially pointed out in a beautifully obvious way that when we know that we are on the brink of changing ourselves fundamentally, we are oftentimes so afraid of the consequences of following our hardwired intuitive guidance ( yes, our soul, heart based guidance), that our head (intellect) will get in the way and stomp down any movement in that direction based on fear of survival or the unknown. At which point…in comes addictive tendencies to numb down our guidance and pacify our discomfort or resistance to the status quo. Doesn't matter what brand of addiction — choose your poison… It's there to stall and prevent the actions necessary to move into greater self-empowerment.
So this begs the question: What does it take to bridge the gap between the parts of ourselves that are so at odds?
A couple things — moving into the process of true congruency requires having a clear voice to speak honestly from a place of personal integrity. It also requires firm boundary development in managing the agenda of one's own life versus the expectations of other people, our familial systems , or the entitlement generated by sociatal imprinting. That's a lot to manage as we try to make decisions and move forward in our lives in ways that serve our evolvement.
Ultimately, however, we each need to start to cultivate an authentic sense of self.
And this is no easy task, by the way, when we haven't been properly modeled, taught or encouraged to even have a sense of who we uniquely are in our growing up. Again, it's our call to look within and "parent ourselves" gently in the process of self-discovery. This can be be hard as hell (btw) and requires us to check in with ourselves often in order to ask: what is it we are being called toward in our lives in the moment? We can only work with what we have in front of us today — nevermind looking at the unknown future. Being in present time in key.
This is rigorous work. Being awake to our process doesn't preclude us from falling on our faces in all different forms:
⇨ betraying ourselves by not asking for what we really want
⇨ avoiding distancing ourselves from toxic situations
⇨ remaining in a sleepwalking state of mind as we busy ourselves to ignore what we know to be true in our lives
We're gonna get in our own way–a lot. But this is part of how we start to respect our own spiritual wiring. We gain respect for ourselves when we start to line up the parts of us that need to be heard, and not dismissed. We gain empowerment when we take action on those life changing course-corrections that both our mind and heart agree upon.
As an aside, the reason I love doing intuitive consultations for people is the simple fact that I'm validating aspects of their soul contracts that they know to be true, yet haven't acted on yet. When we witness the wisdom of our own intutitive selves, that part of us gains strength. Robert Ohotto calls that "soul esteem". I love that term.
And I need to point out that it may be a matter of starting with the "smaller" decisions or changes we're struggling with — as we challenge ourselves to really incorporate a more honest bridging of the heart and mind in certain parts of our lives. I know that this is how I work toward my own process…small victories count!!
Sending lots of love and "soul esteem support" on the journey!


text:
I have tons of stuff in my .bash_profile. The problem is, I use ~3 computers very frequently, and I'm tired of having to copy paste my prefs everywhere. Two of them run Ubuntu 10.10, and one runs OSX. I was wondering if there was a way to use Dropbox, to share a single prefs file. Like, when bash starts, tell it to check
~/Dropbox/Bash/.bash_profile ?
Although, could I also tell emacs to look in
~/Dropbox/Emacs/.emacs somehow?


text:
If you have MS, a few adjustments to your home can help you finish tasks safely and easily. Here's a room-by-room guide:
Get a portable ramp that goes over the threshold of your front doorway so you don't have to go up steps. You can also buy a keyless entry lock and electric door opener. They open the door for you when you press a keypad or remote control.
Inside your house, replace door knobs with lever-style handles. They're easier to turn.
Save your energy when preparing food. Use stools or chairs that are high enough to let you sit at your countertop while you cook or use the sink. You can reduce your meal prep time by using a food processor, blender, or microwave oven.
Look for kitchen aids that make preparing meals easier and safer. Get non-skid mats for counters so your bowls and cutting board won't slide around. You can also buy lightweight cookware that's easy to pick up and carry from the stove to the counter.
These tools can also help with meal prep:
- Utensils with easy-grip handles
- Grabbers to help you reach items on high shelves
- Electric can opener
- Rocker knives to help you cut food with less effort
Living Room
To make your lounging area comfy and safe, get raised chair or lift cushions. They can help you get in and out of your seat more easily.
Avoid throw rugs because they can add to your risk of falling. If you do get them, make sure they are non-skid.
Use extension cords that let you plug in lamps and gadgets without reaching for the outlet, but make sure they're not in the path where you walk.
Hallways and Stairs
Find ways to save your energy when you move to another area of your home:
- Put handrails on both sides of the stairs.
- Get a stair lift if you can't get up and down stairs on your own.
- Install night lights with motion sensors along stairs, hallways, and throughout your home.
A gadget called an ECU (environmental control unit) lets you adjust the heat or air-conditioning with the push of a button. You can also use it to turn on the lights and open and close the drapes.
Some other equipment to consider:
- A bed that raises and lowers by pushing a button
- Button and zipper hooks, a long-handle shoehorn, and sock aids to help you put on and fasten clothing
- A bedside commode so you don't have to walk to the bathroom in the middle of the night
Some simple adjustments can make washing up easier and safer. Look for combs and toothbrushes with easy-grip handles. Also install equipment such as:
- Lever faucet handles that are simple to turn
- Tub bench or shower chair so you can sit while you wash
- Handheld shower head that reaches down to you
- Non-skid surfaces in the shower or tub to prevent falls
Consider making these changes to your toilet:
- A raised or adjustable toilet seat
- A bidet toilet attachment to help you get clean after you use the toilet
- "Bottom Buddy" tool that holds the toilet paper while you wipe and releases it into the toilet after you finish


text:
Classroom Grants
Each year special educators throughout Marin County are eligible to apply for funding to purchase the tools, technology, and teaching materials they need in order to provide their students access to learning opportunities. Since we started our grants program, over $2 million has gone directly into classrooms, and each and every one of our 4000 students per year has benefited.
Much of the technology that has now become standard in our classrooms was pioneered through our Classroom Grants, and we intend to continue with the goal of encouraging new ideas in our classrooms.
Who is Eligible to Apply?
All applicants must be special educators working in Marin County public schools. 100% of their students must have IEP's.
- Special Day Class Teachers
- Resource Teachers
- Speech and Language Pathologists
- Occupational Therapists
New Guidelines
Our guidelines for Classroom Grants have changed. Our new goal is to again provide the sparks for innovation and progress - to encourage new thinking.
- New technologies and innovation are encouraged, especially if they fall within our Four Pillars of Emphasis: Communication, Literacy, Curriculum (adaptation and access), and Technology (using technology to enhance or expand learning opportunities).
- We will no longer purchase equipment that is considered standard for classrooms. This includes replacement of old equipment, and equpiment that is included in any student's IEP. These should be provided by the district, as required by law.
- We will not fund major construction.
- We will not fund experiential grants, including fees for consultants or field trips.
Grant Application Process
Grants applications are open in the fall. Watch for anouncements about the application window


text:
Nanjing City, China; 12, October 2015: ECOWAY ENERGY Engineering Company has elegantly designed infrared heating panels available in various dimensions that can be used to create a warm and healthy environment inside a room during winters. They have customized infrared heating panels that can enhance the value of the home décor and it consumes very less energy as compared to the traditional heating systems. Available in white, black and red colors, these heating panels can be used to give a new meaning to the personalized artworks that adorn walls of the contemporary homes.
The company boasts of its IR Radiator heater that is energy efficient and eco-friendly and doesn't heat up the environment, but creates a healthy and warm way of living for the mankind. This is the reason why they have heating systems for various applications, such as kitchens, bathrooms and yoga studios. One can find several types of glass infrared heating panels and mirror infrared heating panels etc that can help redefine the visual appearance of a bathroom while creating a warm and cozy atmosphere for an enjoyable shower.
The spokesperson of the company reveals that their Glass heater with towel rail is designed for modern bathrooms where one can enjoy a comfortable and healthy way of bath even during chilly winter season. It creates an intense and pleasant warmth feeling and helps floor and walls of the bathroom dry faster. Moreover, the bathroom heating systems are lightweight and are easy to install. Available in several good-looking styles, these bathroom heating panels could be the key to an enjoyable shower and good health.
ECOWAY ENERGY also has an adorable ceiling mounted heater range that can superbly embellish any built space. These mounted heaters are more often used in the yoga studios for creating a comfortable and warm environment for the yoga practitioners. The infrared heating is more suitable for people who want to improve their metabolism, and hence it helps in losing weight. There are several other benefits because of which infrared heaters are used in yoga studios and other places. To know more about these infrared heaters, one may visit the website http://www.ecoart-heating.com.
About ECOWAY ENERGY Engineering Co. Ltd:
ECOWAY ENERGY offers a range of infrared heating systems for domestic as well as public and commercial use. The heating systems are easy to install and can save a significant amount of energy. The company also designs custom infrared heating systems as per the requirements of the clients.
For Media Contact:
ECOWAY ENERGY Engineering Co., Ltd
Email: [email protected]
Website: http://www.ecoart-heating.com


text:
1. Jill Regester of WoodmenLife is a terrific guest blogger who managed to get more comments posted in three days than I have in six months!
2. Billy Beane's message of "challenge accepted norms by being willing to look at everything differently" hit a home run with fraternal leaders.
3. Guy Messick, of Messick & Lauer, spelled out the pros and cons of the fraternal shared services concept being explored by several society leaders in as clear a way as I've ever heard it.
4. I'm sure Ken Frino of A.M Best at times felt like he was on the witness stand during the Spanish Inquisition when explaining the new BCAR rating formula, but members appreciated his willingness to answer every question – even if they did not like some of the answers.
5. If you think your society is exempt from the Department of Labor's new fiduciary rule seriously, you might want to disabuse yourself of that notion – quickly.
6. Fraternal conventions don't have to be stuck in the 1950s! Get rid of the two-tiered head tables and fire up the educational programs that will infuse your members with enthusiasm about what your society can do!
7. Fraternal branding campaign – lots of excitement, many questions to be answered, unlimited potential for building an image that will resonate with consumers.
8. I'd go back to Nashville for a long weekend in a heartbeat. Food, music, and wonderful people – what a combination!
9. There is no better community service partner than the Music Health Alliance! They raised $7,000 on site for their amazing cause, gave attendees the scoop on Nashville hot spots, helped us find local musicians to play the sweetest rendition of the anthems I have ever heard, secured the best Chair's gift ever – a signed tour jacket from George Strait, and sent our staff cookies last week from a company that uses the founder's grandmother's recipe. We are considering making them our permanent partner!
10. Technology IS going to change the way your members and prospects interact with your society. Time to get off the platform and board that train.
11. The Innovation Forum was not only one of the most popular sessions at this year's meeting, it proved that the fraternal system has a bright future if we provide the young professionals in our societies with the opportunities to change the way to "look at everything differently."
12. Folks had strong – and mixed – views about Linda Wertheimer's remarks. But the message I took away was that a disturbingly small percentage of voters decide who gets on the ballot and who gets elected, and perhaps we need to re-think the whole process.
14. The "Diversity" Workshop (Embracing Diversity Among Your Employees and Board Members) should be mandatory for every fraternal C-suite executive and board member. Diversity is not just a moral issue, it's a business ussue. Look for this program to be on the agenda of the 2017 Executive Summit, April 3-5, in Chicago.
15. If you loved Nashville and the Omni Hotel (how about that for a venue!), you are going to be crazy about the Sheraton Wild Horse Pass in Phoenix, the site of our 2017 Annual Meeting. Mark the dates on your calendar RIGHT NOW – September 6-8, 2017 – and, for goodness sake, plan to register early. We've lined up two terrific keynote speakers: Shabnam Mogharabi, CEO and Executive Producer at SoulPancake, an award-winning media and entertainment company that explores 'Life's Big Questions'; and Dr. Robert Cialdini whose book Influence: Science & Practice is a New York Times best seller. Also, next year's meeting will end at noon on Friday, September 8. We know it may be difficult for many of you to get home from Arizona that afternoon, so we're planning an event or two to entice you to stay over another day and enjoy the desert sun. And at $139 per night for a resort like this, how can you afford not to! In fact, why not do what several societies did this year: bring your whole Board of Directors to the Annual Meeting and conduct a Board meeting on the Friday afternoon or Saturday morning following the event?
Filed under: Uncategorized |


text:
New drug effective for rheumatoid arthritis, Stanford scientist finds
STANFORD, Calif. - A new drug appears to offer pain relief and increased mobility to rheumatoid arthritis patients who have exhausted their other medical treatment options. A researcher at the Stanford University School of Medicine led a six-month, multicenter clinical trial that found patients were more than twice as likely to have significant improvement with the new drug than with standard therapy. The findings are reported in the Sept. 15 issue of the New England Journal of Medicine.
The study was a phase-III trial, the final stage of human testing normally required by the U.S. Food and Drug Administration before it will consider approving a drug. Based on the results, an FDA advisory panel met on Sept. 6 and recommended the drug's approval for the treatment of rheumatoid arthritis in cases in which standard therapy has failed.
Abatacept, marketed as Orencia, is made by Bristol-Myers Squibb, which sponsored the study. The lead author, Mark Genovese, MD, Stanford associate professor of medicine (immunology and rheumatology), is a paid consultant for the company.
"This drug works where others haven't," said Genovese, who is also the associate chief of the medical school's Division of Immunology and Rheumatology. "These patients had tried and failed the best therapies to date, and they could still have a very good response to this drug."
Autoimmune diseases such as rheumatoid arthritis are characterized by an overactive immune system that turns its attack to body tissues instead of invading microbes. T cells are immune cells thought to play a major role in the development of rheumatoid arthritis. Abatacept is the first of a class of drugs, called co-stimulation blockers, that selectively impede one of the two signals needed to activate T cells.
"It's exciting to have a therapy with a new mechanism of action," said Genovese. "In the next few years we are going to get an increasing sense of who the best patients are for this therapy and even whether patients with other autoimmune diseases might benefit." Bristol-Myers Squibb is also testing the drug for use in lupus.
Stuart Levine, MD, assistant professor of medicine in the division of rheumatology at Johns Hopkins University School of Medicine, expressed optimism about the new drug. "This would likely be a useful addition to our armamentarium for patients who are failing or have failed first-line therapy," said Levine, who has no financial connections with Bristol-Myers Squibb and was not part of the study.
Rheumatoid arthritis is a disease of the joints that affects more than 2 million Americans. The disease is characterized by an infiltration of the joints with immune cells aimed against the cells lining the joint. The immune system attack causes pain, stiffness and swelling of the joints and can eventually lead to cartilage breakdown, bone loss and weakness of the joints. Without effective therapy, approximately 15 percent of patients will be crippled by their disfigured joints.
Nonsteroidal anti-inflammatory drugs can provide pain relief for rheumatoid arthritis but do not treat the underlying disease. Standard therapy currently includes other agents that suppress the immune system and can actually modify the course of disease. In addition, new medications that block the immune system's response have become available in recent years. Still, many patients don't benefit from these medications.
Bristol-Myers Squibb conferred with Genovese and others to design an international trial to test whether abatacept could benefit these rheumatoid arthritis sufferers who hadn't responded to existing therapy. At 89 sites around the world, 258 of these patients received abatacept as an intravenous injection seven times over the course of the study, and 133 patients received a placebo. Neither the patients nor the physicians knew who received active drug.
After six months of treatment, half of the patients taking abatacept achieved at least 20 percent improvement compared with one of every five taking the placebo. The FDA requires that a majority of patients benefit by this degree before a drug can be approved for treating rheumatoid arthritis.
A 20 percent improvement means a lot, Genovese said, particularly for the severely affected patients in this study, who had an average of 30 tender joints each. "For many of these patients, it means a significant increase in their ability to function," he said. "That means turning on the shower, opening a car door, using the toilet - little actions that make a huge change in their quality of life."
One in five patients taking abatacept achieved at least 50 percent improvement and one in 10 achieved the definition for remission - a difficult hurdle to clear for a group that failed to see significant improvement from all other therapies.
According to the trial results, abatacept appears to be safe, with few side effects. About 2 percent of both groups experienced serious infections. Mild to moderate infections such as bronchitis and upper respiratory tract infections were slightly higher in the group taking the drug - 38 percent compared with 32 percent in the placebo group.
"It's a fine line between trying to modulate the immune system and suppressing it to an extent that you are at an increased risk for infections," said Genovese. The higher risk of less serious infections is something to keep in mind with abatacept.
"Any time you modulate the immune system you are at a potential risk," he said, "but you just have carefully weigh the risks and the benefits.'
Stanford Medicine integrates research, medical education and health care at its three institutions - Stanford University School of Medicine, Stanford Health Care (formerly Stanford Hospital & Clinics), and Lucile Packard Children's Hospital Stanford. For more information, please visit the Office of Communication & Public Affairs site at http://mednews.stanford.edu.


text:
1. Finding a new place to live is kind of a pain, but also reasonably fun. Anyone know of a good source for rental listings aside from craigslist? We're looking for a 2-3 bedroom house around Sunnyvale.
2. I have something funny to tell, but only one of you would find it funny
3. I'm not a NASCAR fan, but i wish i had gone to this weekend's race at Sears Point where Juan Pablo Montoya passed everyone to win his first big NASCAR race
4. I've had two beers tonight, both from the really old box of "beers of the world" that I didn't realize I had until a few weeks ago
5. New product [NAME REDACTED] coming out from work sometime soon is really, really, really wacky. I like it.
6. I have tagged no one for this meme
7. My new DSLR camera is really sweet, though is a lot more of a challenge to use than my old sd200:


text:
Owner: Owyhee Colors
Ontario, Oregon
My love for goats goes back to my school days. My first doe, a black Nubian named Blossom, had me hooked from word go. We raised sheep and goats for many years selling to locals for 4-H and FFA projects. My parents then sold the cattle ranch we grew up on and moved from Baker City area to Ontario. A few years after high school, I moved to Washington and got married. Little did my husband know (because at the time there were no goats to be seen) he had just married "the crazy goat lady." Our home had some acreage and as soon as we got it fenced there were animals on it. One goat quickly became a lot of goats, mostly Nubians and mostly spotted in color. My daughter was born summer of 2010 and the need to be closer to family brought us to Oregon. We quickly learned that the demand for the dairy does was much less, yet I did not want to give up my colored does. It was then, while searching for a Boer buck, we discovered the dapple and spotted colored Boer goat…we have been hooked since. Our first SRMGA Production Sale was in the fall of 2013 where we had hopes of taking home the only spotted buck in the sale. We were not so lucky to get him but, did come home with two nice bucks and a couple fullblood does and thus began a partnership with my parents and my husband and I we call Owyhee Colors. We strive to breed a great colored goat that will perform well in the field giving you great mothering instincts and milk. We are also looking for one that will compete in the ring. We have a lot of true percentages as they were the foundation for where we began and great does are hard to give up no matter what the breed. We have met some amazing new friends and people we would call family through this association and its shows. As that first time showman, in an environment unfamiliar to us, it was great to be welcomed with open arms and have people willing to help you with whatever you needed. It is my goal to show that same compassion and willingness to help to others while serving as an SRMGA Board member. I look forward to many great events, meeting and seeing new and old faces.