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Griffin: Ultimate Immunoprecipitation Guide
When performing an IP experiment, then consider to run a preliminary western blot first in order to get a feel for what the antibody is capable of binding.
RIPA (Radio ImmunoPreciptation Assay) buffer is a traditional name for an array of recipes that have found success over the years. Below are details that can contribute to optimizing immunoprecipitation experiments. Lysis buffer components can and will influence the efficiency of the IP reaction. Detergent composition is a major factor for IP reactions. Adjusting salt concentration and detergent composition will influence the efficiency of the IP reaction. Membrane bound proteins (proteins based in lipid rafts), protein complexes, and protein charge can all influence the efficient yield of a gene product isolate within the lysate prep.
- The amount of protein present in cells will vary with cell type. 1 to 10 million cells can yield ~1mg of protein.
- 70% confluent 10 cm dish can yield ~600-1000 ug total protein
- Certain types of proteins (ie insoluble, modified) may escape classical lysis and purification procedures under the mild conditions outlined below. Additional methods that may improve on enrichment include cross-linking and purification under denaturing conditions.
Common RIPA components
PBS : Salt prevents non-specific protein aggregation
Tris-HCl : Buffering agent prevents protein denaturation
NaCl : Buffering agent prevents protein denaturation
1% Nonidet P-40 or Igepal CA-630 : Non-ionic detergent to extract proteins, form lipid micelles
1% Triton X-100 : Non-ionic detergent to extract proteins, form lipid micelles - to use in place of Nonidet/Igepal
0.5% sodium deoxycholate : Ionic detergent to extract membrane protein and isolate lipids
0.1% SDS : Ionic detergent to extract membrane protein and isolate lipids
PMSF : Stock Solution 100 mM (100x stock) or 200 mM in Isopropanol; use @ 1 mM. Store PMSF solution up to 6 months @ 4C.
EGTA : Protease Inhibitor, Prevents protein degradation. You can make your own, or several vendors have convenient crushable pills that form a protease inhibitor cocktail solution.
Na3VO4 (Sodium Orthovanadate) : Tyrosine phosphatase Inhibitor; hydrostatic interference of active sights of phosphatases
NaF (Sodium Fluoride) : Serine/Threonine phosphatase Inhibitor; hydrostatic interference of active sights of phosphatases
Phosphatase Inhibitors
Phosphorylation/dephosphorylation of proteins influences hydrostatic relationships. Proteins undergo covalent attachment of a phosphoryl group (phosphorylation) at serine, threonine, or tyrosine residues. Phosphate groups are removable via protein phosphatases. During the extraction of phosphorylated proteins from cell and tissue, there are advantages to preserving the phosphorylation states of total protein.
- (-)-p-Bromotetramisole oxalate
- Cantharidin
- Calyculin A
- Discodermia calyx
- Imidazole
- Sodium Fluoride
- Sodium Molybdate
- Sodium Orthovanadate
- Sodium Tartrate Dihydrate
Protease Inhibitor Cocktails
Tableted formulations containing water-soluble protease inhibitors are user-friendly and effective substitutes in lysis buffers. When using divalent/trivalent columns for enrichment of phosphoproteins, be sure to avoid EDTA/EGTA containing cocktails.
- S8820: Sigma SIGMAFAST™ Protease Inhibitor Tablets
- 78429: PIERCE Halt Protease Inhibitor Cocktail (100X)
- 04693116001: Complete™ Roche Protease Inhibitor Cocktail Tablets
- sc-29131: Santa Cruz Biotechnology Inc.,Protease Inhibitor Cocktail Tablet, EDTA-free
Protease Inhibitor components
- Aprotinin
- Bestatin
- Calpain Inhibitor I & II
- Chymostatin
- Leupeptin
- Alfa-2 Macroglobulin
- Perfabloc SC
- Pepstatin
- TLCK-HCl
- Trypsin Inhibitor (chicken egg white, soybean)
Protein A/G/L Agarose
Protein A & Protein G bind to most mammalian immunoglobulins primarily through their Fc regions. Protein L is a kappa light chain specific Ig-binding protein.
Protein A/G/L are common to covalently couple by cyanogen bromide to highly cross-linked 4% agarose beads. This type of matrix is stable in most aqueous buffers.
- Typical Ligand density: ~3 ug Protein/ul of bead
- Binding capacity: ~15-20 ug Ig/ul bead
- Bead structure: 4-6% cross-linked agarose
- Bead diameter: 40-170 um
- Temperature stability: 4-40 C
Protein A
Native protein A is a single chain (predicted 42 kDa, SDS-PAGE 46 kDa), glycoslyation-free, cell wall component produced in strains of Staphylococcus aureus. Protein A binds specifically to the Fc region of immunoglobulin molecules, including IgG. Protein A has four high-affinity (Ka = 108/mole) binding sites toward Fc region of IgG of several species (two sites can bind at a time). Protein A is heat-stable and retains conformation even after exposure to denaturing reagents such as 4 M urea, 4 M thiocyanate and 6 M guanidine hydrochloride.
Protein G
Native protein G is a bacterial cell wall protein from group G Streptococci that contains two immunoglobulin binding sites, an albumin binding site, and cell surface binding sites. Native Protein G. The recombinant form of Protein G (predicted 17-21 kDa; SDS-PAGE 31-34 kDa) contains only the two immunoglobulin binding sites to reduce nonspecific binding when purifying immunoglobulins.
Protein L
Native protein L is an kappa light chain specific Ig-binding protein that originates from the bacteria Peptostreptococcus magnus. Protein L binds Igs through interactions with the light chains. Because no part of the heavy chain is involved in the binding interaction, Protein L binds a wider range of Ig classes than Protein A or G. Protein L binds to representatives of all classes of Ig, including IgG, IgM, IgA, IgE and IgD. Single chain variable fragments (ScFv) and Fab fragments also bind to Protein L.
In humans and mice, kappa (k) light chains predominate. The remaining immunoglobulins have lambda (l) light chains. Furthermore, Protein L is effective in binding only certain subtypes of kappa light chains. For example, it binds human VkI, VkIII and VkIV subtypes but does not bind the VkII subtype. Binding of mouse immunoglobulins is restricted to those having VkI light chains.
Protein L matrix is suitable for purification of kappa light chain-containing monoclonal antibodies from ascites or culture. Protein L is useful for purification of VLk-containing monoclonal antibodies from culture supernatant because it does not bind bovine immunoglobilins, which can be present in media. Protein L does not interfere with the antigen-binding site of the antibody.
Chicken IgY
Protein L does not react with chicken IgY light chains; PMID 15857176
RIPA Buffer Recipes
There are 3 different RIPA style lysis buffers outlined below including a brief summary of their purpose.
These may be made in large volumes. Add inhibitors fresh at time of use from stock solutions
Lysis Buffer 1:
A tried and true lysis buffer for most signaling intermediates and soluble/cytosolic factors.
- 1x PBS
- 1% Nonidet P-40
- 0.5% sodium deoxycholate
- 1 mM Na3VO4 Sodium orthovanadate activation
- 10 mg/ml PMSF (200 mM) in isopropanol (add at 10 µl/ml RIPA)
- Protease inhibitor cocktail
Aprotinin activity is measured by KIU (KIU = Kallikrein Inhibitory Unit) Since the vial contains other components which makes the total dry. I recommend the following procedure to be used with this product: The normal working concentration range for aprotinin is either 0.5ug-2ug/ml (protein weight/volume) or 10 KIU-100 KIU/ml (units/volume). 1ug aprotinin/ml of RIPA buffer works well.
Lysis Buffer 2:
An SDS free lysis buffer to consider with Co-IP approaches. Used this one in graduate school to IP EGFR effectively.
- 150 mM NaCl
- 50 mM Tris-HCl pH 7.4
- 1% Nonidet P-40
- 0.25% Sodium Deoxycholate
- 1 mM EGTA
- 1mM PMSF
- Protease inhibitor cocktail
- 1 mM Na3VO4
- 1mM NaF
Lysis Buffer 3:
This recipe calls for Brij 35 which is a non-ionic detergent, great for dissociating membrane complexes and essentially much gentler than SDS. This is also a great lysis buffer for phospho-proteins.
- 10 mM KPO4 (phosphate buffer)
- 1 mM EDTA (chelate)
- 5 mM EGTA (chelate)
- 10 mM MgCl2 (chelate)
- 50 mM †-glycerophosphate (inhibits serine-threonine phosphotase activity)
- 0.5% NP-40 (stabilizer of proteins/enzymes)
- 0.1% Brij 35 (non-ionic detergent)
- 0.1% deoxycholic acid (non-ionic, non-denaturing detergent)
- 1 mM sodium orthovanadate (inhibits tyrosine phosphotase activity)
- Protease inhibitor cocktail
- Roux PP, Richards SA, and Blenis J. Phosphorylation of p90 ribosomal S6 kinase (RSK) regulates extracellular signal-regulated kinase docking and RSK activity. Mol Cell Biol. 2003 Jul;23(14):4796-804. DOI:10.1128/mcb.23.14.4796-4804.2003 |
Immunoprecipitation/Co-IP enhancement strategies
Covalent Cross-Linker
Cross linkers represent a shotgun strategy that captures interaction complexes by chemical crosslinking of intracellular proteins prior to cell lysis and immunoprecipitation.
- DSP (dithiobis(succinimidyl propionate) CAS Number 57757-57-0): Membrane-permeable, Bifunctional protein cross-linker that contains amine-reactive N-hydroxysuccinimide (NHS) esters. NHS esters react with primary amines at pH 7-9 to form covalent amide bonds. NHS-ester crosslinking reagents target primary amines of lysine (K) residues and N-terminus. Lysine residues are generally abundant and accessible on the hydrophilic phase of proteins, and cross-link with high efficiency.
- Use fresh tissue culture-grade DMSO when using a crosslinker on living cells as unsealed/opened DMSO absorbs atmospheric water, and will pre-react with the cross-linker
Detergent % Optimization
Titration of detergent composition with respect to sample type (ie cell lineage, species, tissue type) improves enrichment of lipophilic components, and improves total soluble fraction yield.
- 0.1%-1% Nonidet P-40 or Igepal CA-630 : Non-ionic detergent to extract proteins, form lipid micelles
- 0.1%-1% Triton X-100 : Non-ionic detergent to extract proteins, form lipid micelles - to use in place of Nonidet/Igepal
- 0.1%-1.5% sodium deoxycholate : Ionic detergent to extract membrane protein and isolate lipids
- 0.05%-0.1% SDS : Ionic detergent to extract membrane protein and isolate lipids
Adherent cell sample preparation
Below is a procedure for adherent cells (ie A431, A549, Hela, NIH3T3)
- Remove culture medium and rinse a subconfluent, 100 mm cell culture plate (80% confluent plate yields ~600-1000 ug protein total) with PBS at room temperature. The following steps should be performed on ice or at 4° C using fresh, ice cold buffers.
Optional: For monolayer cells, do a trypsin treatment to lift cells off the flasks prior to adding the RIPA buffer, instead of scraping the cells for a more gentle approach. If you are running a time course experiment, this is not feasible since the cells must be arrested and lysed immediately.
- Add 0.8 ml of ice cold fresh RIPA buffer to the 100 mm cell culture plates OR 0.5 ml per 5 x 10e6 cells/60 mm dish.
- Gently rock plates for 15 minutes at 4° C or let the plates set on ice. This step will allow the lysis buffer to act on the cells and will increase the total yield of soluble protein. Scrape the adherent cells with a cell scraper and then transfer the scraped lysate into a sterile microcentrifuge tube. Place the tube on ice.
Optional: wash the plate once with 0.2 ml of RIPA buffer and combine with first lysate. When running multiple plates this can be tedious and not necessary if enough attention is given to the initial harvest.
Optional: Add 10 µl of 10 mg/ml PMSF stock to the lysate. If a protease inhibtior cocktail is used fresh with the RIPA buffer, this is not necessary.
- Sonicate each sample on a 70% duty cycle or less by placing only the very tip of the pin into the vial, then slowly lowering it into the lysate until it foams completely and then stop. Alternatively, pass the lysate through a 21 gauge needle to shear the DNA & incubate 30–60 minutes on ice.
- Microcentrifuge cell lysates at 12,000xg for 15 minutes at 4°C.
- The supernatant fluid is the total cell lysate. Transfer the supernatant to a new microfuge tube and discard the pellet. Quantitate the protein amount by Bradford or BCA.
Suspension cell sample preparation
- Collect approximately 2.0 x 107 cells by low-speed centrifugation (e.g. 200xg) at room temperature for 5 minutes. Carefully remove culture medium.
- Wash the pellet with PBS at room temperature, and again collect by low-speed centrifugation. Carefully remove supernatant.
- Add 1.0 ml of ice cold RIPA buffer with freshly added (Protease Inhibitors) and/or (Phosphatase Inhibitors). Gently resuspend cells in RIPA buffer with a pipet and incubate on ice for 30 minutes.
- Further disrupt and homogenize cells by hydrodynamic shearing (21-gauge needle), dounce homogenization or sonication, taking care not to raise the temperature of the lysate. (Optional: Add 10 µl of 10 mg/ml PMSF stock; sc-3597) Incubate 30 minutes on ice.
- Transfer to microcentrifuge tube(s) and centrifuge at 12,000xg for 15 minutes at 4° C. The supernatant fluid is the total cell lysate. Transfer the supernatant to a new microfuge tube. This is your whole cell lysate. For increased protein recovery, resuspend the pellet in a small volume of RIPA, centrifuge and combine supernantants.
- The supernatant fluid is the total cell lysate. Transfer the supernatant to a new microfuge tube and discard the pellet. Quantitate the protein amount by Bradford or BCA.
Tissue extract sample preparation
There are a few approaches to optimizing protein yield from whole tissue extract. For whole animal studies, arresting the covalent modification state of the entire proteome is essential to obtaining accurate data about the treatment or phenotype effect on the tissue being extracted.
1) Immediately liquid nitrogen flash freeze tissue/organ then stored at -80 C
2) Immediately heat denature the organ from the sacrificed animal by microwave in a sealed container
- Weigh tissue and dice into very small pieces using a clean razor blade. Frozen tissue should be sliced very thinly and thawed in RIPA buffer containing (Protease Inhibitors) and/or (Phosphatase Inhibitors). Use 3 ml of ice cold RIPA buffer per gram of tissue.
- Further disrupt and homogenize tissue with a dounce homogenizer or a sonicator, maintaining temperature at 4° C throughout all procedures. (Optional: Add 30 µl of 10 mg/ml PMSF (sc-3597) stock per gram of tissue.) Incubate on ice for 30 minutes.
- Transfer to microcentrifuge tubes, centrifuge at 10,000xg for 10 minutes at 4° C. Remove supernatant and centrifuge again. The supernatant fluid is the total cell lysate. A longer centrifugation may be necessary to obtain a clear lysate.
- The supernatant fluid is the total cell lysate. Transfer the supernatant to a new microfuge tube and discard the pellet. Quantitate the protein amount by Bradford or BCA.
Immunoprecipitation Procedure
I) Incubate cell lysate (500-1000 ug) with (2-5 µg) primary antibody (optimal antibody concentration should be determined by titration) for 2 hours at 4°C.
II) Add 20 µl of appropriate agarose conjugate suspension (Protein A-Agarose, Protein G-Agarose, Protein A/G-Agarose or Protein L-Agarose).
Protein A-Agarose : specific binding to mouse IgG2a, IgG2b and IgA, rabbit polyclonal Abs, human IgG1, IgG2 and IgG4
Protein G-Agarose : specific binding to mouse IgG1, IgG2a, IgG2b, IgG3, rat IgG1, IgG2a, IgG2b and IgG2c, rabbit and goat polyclonal Abs, human IgG1, IgG2, IgG3 and IgG4
Protein L-Agarose : specific binding to mouse, rat and human IgG, mouse and human IgM, IgE and IgA proteins and scFv and Fab fragments
What is Protein G PLUS? Protein G PLUS is a genetically engineered form of streptococcal Protein G that has an increased capacity and has had the albumin binding site removed to reduce background.
III) Cap tubes and incubate at 4 C on a rocker platform or rotating device for 2 hour to overnight.
IV) Collect pellet by centrifugation at 2,500 rpm (approximately 1,000xg) for 5-10 seconds. A touch spin will work. With enough samples, gravity will pellet the beads as well.
V) Carefully aspirate and discard supernatant. The trick here is to slowly aspirate with the needle touching just the top of the liquid and slowly draw down so that the needle is pulling at the surface tension of the supernatant. This will ensure no loss of beads.
VI) Wash pellet 3 times with either RIPA buffer (more stringent) or PBS (less stringent), each time repeating centrifugation step above.
VII) After final wash, aspirate and resuspend pellet in 40 µl of 2x electrophoresis sample buffer. Or elute proteins with an appropriate antibody elution buffer.
VIII) Boil samples for 2 minutes. Load sample.
Related Recipes
Electrophoresis sample buffer (2x): Mix 1.0 ml glycerol, 0.5 ml 2-mercaptoethanol, 3.0 ml 10% SDS, 1.25 ml 1.0 M Tris-HCl, pH 6.7, and 1–2 mg bromophenol blue. Store frozen in small aliquots. Alternatively, make buffer without 2 -mercaptoethanol and store at room temperature. Add 2-mercatoethanol just before using.
Sample buffer formulation:
- 7 ml Tris·Cl/SDS, pH 6.8
- 3.0 ml glycerol (30% final)
- 1 g SDS (10% final)
- 0.93 g DTT (0.6 M final)
- 1.2 mg bromphenol blue (0.012% final)
- Add H2O to 10 ml
- Store in 0.5-ml aliquots at -70°C
Latex Agglutination Assay
The latex agglutination assay is a laboratory method to check for presence of antibodies or antigens in bodily fluids;saliva, urine, cerebrospinal fluid, or blood. The test depends on what type of sample is needed.
Samples are sent to a lab, where it is mixed with latex beads coated with the specific antibody or antigen. If a affinity reactive substance is present, the latex beads will clump together (agglutinate).
For example, a child has strep throat, a throat swab is taken. The sample is mixed with latex beads that are coated with antibodies against the bacteria. The bacteria in the sample will react with the antibodies on the latex particles causing clumping (agglutination).
Latex agglutination results take about 15 minutes to an hour.


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Heidi Klum made a habit of regularly renewing her vows with Seal during their seven year marriage, but since calling it quits with the "Kiss from a Rose" singer in January, Klum isn't as open to walking down the aisle again.
"I don't know if I'll get married again," the 39-year-old mom of four told German newspaper Bild am Sonntag three months after she filed for divorce from Seal, citing "irreconcilable differences." (Though she filed papers in April, the couple confirmed their split in January 2012.)
"Although I really enjoyed being married. I'm a dreamer," she continued. "I loved our annual fairy tale weddings that were our big family parties. But sadly, it somehow didn't work out."
Prior to announcing their split, the couple — who have four children: Leni, Henry, Johan, and Lou — renewed their vows in May 2011 in a masquerade-themed affair in Palm Beach, Fla.
Now that she's on her own, the single Project Runway hostess credits her broken relationship with giving her a new perspective on life.
"I think everyone should work on their dreams. I try to. I look into the future, concentrate on my family and I have to say that I'm happy," reasons Klum. "There are worse things in life than a separation."
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subset: commoncrawl


text:
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1 suppliers match your filters. Clear
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text:
LONGMONT, Colo.--(BUSINESS WIRE)--
UQM TECHNOLOGIES, INC. (NYSE MKT: UQM), a developer and
manufacturer of power-dense, high-efficiency electric motors, generators
and power electronic controllers, will hold a conference call with
members of the investment community on Thursday, May 11, 2017, at 4:30
p.m. Eastern Time. To participate in the call, dial 1-888-241-0326
approximately 10 minutes before the conference is scheduled to begin and
provide conference ID code "14076403" to access the call. International
callers should dial +1-647-427-3411.
Joseph R. Mitchell, UQM Technologies' President and Chief Executive
Officer, and David I. Rosenthal, the Company's Chief Financial Officer,
will be reviewing the Company's operating results for the quarter ended
March 31, 2017.
For anyone who is unable to participate in the conference, access to a
recording will be available for seven days following the call. Dial
1-855-859-2056 and enter replay access code 14076403# to access the
playback. International callers should dial +1-404-537-3406. Please
allow two hours from the time of the conference call for initial setup
before access.
About UQM Technologies, Inc.
UQM Technologies is a developer and manufacturer of power-dense,
high-efficiency electric motors, generators, power electronic
controllers and fuel cell compressors for the commercial truck, bus,
automotive, marine, military and industrial markets. A major emphasis
for UQM is developing propulsion systems for electric, hybrid electric,
plug-in hybrid electric and fuel cell electric vehicles. UQM is TS 16949
and ISO 14001 certified and located in Longmont, Colorado.
Please visit www.uqm.com
for more information.
Source: UQM Technologies, Inc.
View source version on businesswire.com: http://www.businesswire.com/news/home/20170504005335/en/
Source: UQM Technologies, Inc.
EnergyTech InvestorShawn Severson, 415-233-7094orUQM
Technologies, Inc.David Rosenthal, 303-682-4900
Danfoss Editron (UQM)
4120 Specialty Pl.
Longmont, Colorado 80504


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Be the first property to be listed on this page - instant online activation.Click here to register >
Ken Smith, Property 12069
You may want to consider these holiday rentals in other areas of Black Sea Coast.
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The Kamelia Villa is a spacious one bedroom property comfortably sleeping up to a family of 4. A large patio gives out onto the pool area.
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Our newly completed furnished and air conditioned Studio Apartment offers great value and good quality accommodation on the spacious excellent family complex of Nessebar Fort Club in the Bulgarian Black Sea Resort of Sunny Beach.
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On offer are two villas for rent situated in the tranquil area of General Kantadzhievo. The area (being a natural reserve called Batova gora) is very picturesque and suitable for hiking among the virgin nature, fishing, and hunting a variety of game.
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Apartment is well furnished for a great self catering holiday, internet can be paid for locally so take your laptop, pool & sunbeds free, BBQ available for hire on the poolside, but complex must be quite after 22.00hrs. Around 15 minutes walk.
Spacious & modern apartment with spectacular sea views. Modern & well furnished on the beach. All facilities of a quality Aparthotel onsite. Sea view terrace for bedroom & lounge. Sleeps 4 comfortably. Restaurant, bars & leisure all onsite.


text:
Pollen analysis - Les Avenières (F-38), L'Eau morte ou Eaux-mortes
Disciplines : archaeology environmental sciences geography, physical ...
Collected date : 2010
Creators : Emilie Gauthier
Natural site: drilling in an ancient meander of the Rhône river. 17 palynological samples. Site included and completed in the HDR of Emilie Gauthier (2012).


text:
Few reasons why a lot of girls love Edward Anthony Masen Cullen.
A normal guy would say:
"I love you, baby!"
Edward would say:
"You are my life now."
As you leave the house, a normal guy would say:
"Bye. See ya!"
Edward would say:
"Hurry back to me."
While you've gone far away, a normal guy would say:
"I miss you."
Edward would say:
"It's like you've taken half myself with you."
If you died, a normal guy would find another.
Edward would kill himself because life without you isn't worth living.


text:
Fire burning along Warrego Hwy
FIRE crews are battling a blaze along the Warrego Highway.
Three trucks were called to the scene, just before the Laidley turnoff where a small area of grass is burning on the southern side of the highway.
A Queensland Fire and Rescue spokesperson said while there may be a lot of smoke in the area the fire, reported at 1.15pm, had been brought under control.
Drivers in the area have been reminded to be cautious and drive to conditions.
Earlier this month Ipswich residents were warned this season predicted to be particularly unpredictable with high fuel loads across the region.
In particular grass fires are a major concern for Rural Fire Services Queensland area director Paul Storrs who says the region has been classed as having a Very High Fire Danger risk.
Unseasonal wet weather during winter has made hazard reduction burning difficult throughout parts of Queensland, meaning Ipswich residents must be more prepared than ever for disaster to strike.
"People with plans are the ones who survive," Mr Storrs said earlier this month.
"Land owners need to know if they will stay and defend or leave and prepare for that.
"Right around the district we are reminding people to clean out their gutters, move firewood away from houses and make sure smoke alarms are working."
For more on how to prepare for bushfires, visit ruralfire.qld.gov.au.


text:
We have been trading from our shop in Bishop's Castle, Shropshire since 1997, stocking a range of original items from the 1890s to the 1930s, particularly Art Deco.
We specialise in art glazed pottery, art glass, metal ware, costume jewellery, evening bags, early plastics, lighting and small furniture. We also sell a small selection of reference books covering some of the above areas. Usually the products of commercial manufacturers, the items we stock are often unique and interesting for the decorative effects achieved.


text:
There are cars that are fun to drive, and then there are cars that, because you know you're a goner if you crash, are just scary to drive. But since most structurally unsound beaters aren't exactly capable of exciting speeds, fast and scary are often mutually exclusive. This, the Brilliance FRV 1.8T, may very well be an exception.
If you remember the significantly less than five-star crash test of the Brilliance BS6, you may never want to so much as ride in a Brilliance, let alone drive one quickly. But the FRV, also known as the Brilliance BS2, will now be offered with a 1.8-liter turbocharged engine connected to a 6-speed manual gearbox. So, as if being terrified of crashing wasn't enough, now you'll be terrified while going much faster. Which, actually, sounds quite thrilling.


text:
750 Peyton St, Barboursville, West Virginia 25504
Senior Care Helper aims to be the first source in senior directed care. With a transparent edge and a growing library of information regarding senior care options, Senior Care Helper is well on its way to being a leader in the Senior Care community.
Important: Senior Care Helper is in no way affiliated with any of the listings provided on this site. We do our best to connect families with wonderful caring companies; however, we are unable to discern the quality or value of any of the providers listed.


text: PORTLAND, Ore. -- Suzann Pettersen won the Safeway Classic for the second time in three years Sunday, taking advantage of playing partner Yani Tsengs final-round collapse at Columbia Edgewater. Three strokes behind leader Tseng entering the round, the third-ranked Pettersen closed with a 5-under 67 for a two-stroke victory over second-ranked Stacy Lewis. Pettersen finished at 20-under 268 for her 12th LPGA Tour victory. The 32-year-old Norwegian also won the LPGA Lotte in Hawaii in April and won a Ladies European Tour event in March in China. She won the 2011 tournament at Pumpkin Ridge. "I felt like this was a good bounce back from last week when I kind of screwed it up on the last day," said Pettersen, who finished eight strokes behind winner Lydia Ko in the Canadian Womens Open after beginning the final round a shot back. "I tried to give myself chances. Once I got going I felt like I made a lot of clutch putts. I guess thats what made me win today. The putter was a good friend of mine." Tseng followed her third-round 63 with a 78 to tie for ninth at 12 under. The Taiwanese star, a 15-time winner on the tour who had a 109-week run at No. 1 in the world ranking, is winless in 37 events since the Kia Classic in March 2012. Lewis, a three-time winner this year, shot a 68. "It was a little frustrating. I played really good, though, so I cant be upset," Lewis said. "A lot of those putts I hit exactly where I wanted to and they just didnt go in. A bogey-free tournament Ive never done before, so I keep checking things off the list." Lizette Salas was third at 17 under after a 69, and 2008 champion Cristie Kerr was another stroke back after a 69. A 72-hole event for the time, the tournament returned to Columbia Edgewater after four years at Pumpkin Ridge. Columbia Edgewater also was the tournament site from 1990-2008. For Pettersen, the Safeway victory capped an emotional three-week period. It started with Solheim Cup, where Pettersen helped Europe rout the United States in Parker, Colo. "Seems like Portland is a good stop for me," Pettersen said. "Seems like I always play well here. This was a good finish to a solid three-week stretch for me." A key point during Pettersens final round was her response after hitting a poor 8-iron at the par-3 second hole that led to a double bogey. "I felt like an amateur, to be honest. That was a disaster of a hole," Pettersen said. "I decided before I went out (Sunday) just not to have too many reactions, good or bad. I knew there were birdies out there if you really get it going." Pettersen quickly with a birdie on the par-4 third that started a stretch where she birdied five of the next six holes and eight of 11. Pettersen made an 18-foot birdie putt at the par-3 eighth to overtake Lewis for the lead, curled in a 5-footer for birdie on the par-5 10th and made a 25-footer from off the green on the par-4 11th. Pettersen opened up a four-stroke lead with an 8-foot birdie putt on the par-3 13th. She ran into trouble at the 175-yard, par-3 16th, following a short tee shot with a poor chip. But she minimized the damage when her next chip rolled within 3 feet to set up a bogey-saving putt. Pettersen said she didnt know she had a three-stroke lead on the 16th tee. "It was a pretty bad swing. Even worse chip. But it was a good 4," Pettersen said. Pettersen closed out the tournament with two pars. She was greeted on the 18th green by Sophie Gustafson, the longtime European Solheim Cup player who announced her retirement from the LPGA Tour on Friday. "We have a history together and to see her on the 18th was a little bit special," Pettersen said. "I never thought she would kind of stick around. When she says she is done, shes done. She is usually out of there. One thing is to get to know each other on the golf course, but she has become a good friend of mine and thats what you do for friends." Discount Nike Free Shoes NZ .DeMarco Murray got his 20th carry late in the fourth quarter with a 10-point lead, not long after Dez Bryant made his final catch of the day.Sure enough, a 44-17 win over Washington didnt change Dallas post-season position. Nike Free Shoes Wholesale NZ . So much so that even a simple foul pop up to the first baseman turned into a run. Houstons four-run rally in the ninth inning to beat the Seattle Mariners 6-4 on Monday night was capped by a bizarre play that started as Crowe fouled out to first baseman Justin Smoak with runners on second and third. http://www.nikefreenz.com/. -- Once again, Carlos Santana was a huge hit in Kansas City. Nike Free Shoes NZ . Sopoaga hit the upright with his first shot at goal from 15 metres. He then kicked nine goals in succession -- two conversions and seven penalties -- before being replaced in the 62nd minute, three points short of the Highlanders record for most points in a match. Cheap Nike Free NZ . "Opinion: Womens World Cup is the best Soccer of the year," Hanks tweeted to his 8.73 million followers on Friday. "Hey FIFA, they deserve real grass. Put in sod.EAST RUTHERFORD, N.J. -- Restricted free-agent linebacker Mark Herzlich has re-signed with the New York Giants. The Giants announced the signing of the fourth-year player Friday without releasing terms. In his first three years with the Giants, Herzlich played in 43 games with six starts. He has 57 tackles (44 solo) on defeence and 33 special teams tackles.dddddddddddd The former Boston College product played in all 16 games for the second year in a row, with two starts. He had a career-high 32 tackles (26 solo) and a team-leading 14 special teams tackles (12 solo) playing mostly as a backup to Jon Beason at middle linebacker. 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Dark Waters
100% Synch Requirements
- Successfully braced 5 times when attacked.
- Don't hit any rocks in the minefield.
- Destroy 80% of Washington's armada in the upper bay.
When you begin, you'll be heading against the wind in the direction of four ships. Three large and one small. The first ship you'll face will be to the most right. As you approach this ship, it will steer to your left to hit you with it's canons. Pass on the right for the counterattack. After this initial passing, the ships will scatter. The smaller can easily be destroyed by hitting it with your smaller cannons.
Once all four ships have been eliminated, your map will update to a wreckage. Connor's crew will be ordered to search this wreckage to steal fly the King's colors in order to make it into New York.
As you move through to the next area, there will be mines in the water that will explode by touching them with your ship or shooting them with your cannons. Try your best to hug the rocks without touching them. Hitting rocks will fail 100% synch. There is plenty of room between the mines and the rocks that allow for your ship to pass through. Be mindful of rogue winds during this part. Keep your ship at half sail for better control and tighter turns if need be.
As you approach the smaller ships, you'll be notified by Faulkner to blow the ships up. Aim your smaller cannons at the nearest mine to cause a chain explosion that destroys all the smaller ships blocking your way through.
Next you'll be bombarded by a fleet. You can either stay to fight them or head forward to the stationary ships on the other side of the water. Ignoring the other ships will cause 100% Synch failure.
When the cutscene ends, the wind will be in your favor. Toss the ship into full sail and head straight for the Man-of-War to ram it. The only defense you'll have from incoming fire is to brace when attacked.
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My serial novella, The Darkest Shade Of Grey, is being published in four weekly installments by The Red Penny Papers. It's free to read online, so get on over there and check it out.
Episodes 3 and 4 will be published over the next two weeks.
If you enjoy it, please do share the links with your family, friends and colleagues.


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Electromagnetic fields of all frequencies represent one of the most common and fastest growing environmental influences, about which anxiety and speculation are spreading. All populations are now exposed to varying degrees of EMF, and the levels will continue to increase as technology advances.
Electric and magnetic fields (EMF) are invisible lines of force that surround any electrical device. Power lines, electrical wiring, and electrical equipment all produce EMF. There are many other sources of EMF as well.
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Check Out All the Types of Windows to Choose From Before Deciding Upon One for Your Home Construction Project
By Mark J. Donovan
Though the double hung window maybe arguably the most popular window style today there are still many other window styles you should consider when building your new home or home addition.
Double Hung Window
A double hung window consists of two vertical panels that both can be moved up or down. The double hung window is one of the most popular windows installed today due to its flexibility in window panel opening.
Single Hung Window
The single hung window is akin to the double hung window, however only the bottom panel moves. The top window panel is fixed.
Casement Windows
Casement windows are still fairly popular. They normally come in either single or pairs, and one or both panels are hinged and can be opened via a crank located most commonly near the bottom of the window.
Fixed Windows
Fixed windows come in all shapes and sizes including circles, squares, triangles, and trapezoids. They are ideal for letting in natural light without the hardware concerns common with other windows. They are often used as accent pieces for both the inside and outside of the home.
Bay Windows
Bay windows are still very popular in homes today. They are often used in large rooms such as in family rooms, dining rooms, and living rooms. Due to their large size and two dimensional shape, they allow much more light to enter a home all throughout the day. Often bay windows have a fixed panel in the center and either double or single hung windows on the sides.
Bow Windows
Bow windows are akin to bay windows in the sense that they let much more light in throughout the entire day. They typically utilize more glass panels to achieve the curved or bow look.
Jalousie Windows
Jalousie windows consist of many thin strips of glass stacked vertically upon one another and that are louvered. They can be cranked opened or closed, similar to a venetian blind. Often they are used in three season rooms or porches.
Awning Windows
Awning widows are single panel windows that are hinged at the top of the window frame. The window can be pushed or cranked open at its base. They are ideal for allowing air to enter a home even on rainy days. Due to the fact that they are hinged at the top of the window, no rain can enter into the home.
Sliding Sash Windows
With sliding sash windows the window panels slide open horizontally. One or more panels can be designed to slide along a track. They are ideal for smaller rooms or in kitchens where it may difficult to open another window type.
So before choosing the ubiquitous double hung window for your new custom home or home addition project take a closer look at all the other window types out there on the market.
Thursday, August 02, 2012
Types of Window Styles
Check Out All the Types of Windows to Choose From Before Deciding Upon One for Your Home Construction Project
Posted by Unknown at 12:20 PM 0 comments


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Your cart is emptyShop
Collège National Orthodoxe in Tripoli, Lebanon is one of the first schools in the country that has implemented the mozaLearn educational system and the mozaBook presentation software in its classrooms. The IT team of the prestigious school installed mozaBook on interactive smartboards in the classrooms and received instant appreciation from both teachers and students.
What's more, the media crew of the institution prepared a superb demo video as well, detailing the most important features of the system!
Michel Katra, the principal of the school says,
"As our students are learners of the digital era, we needed a system to make everything much more interactive and engaging for them. The mozaBook software is a state-of-the-art solution in education and we wanted to be among the first schools in Lebanon to adopt this system."
While Mr. Kilo (Physics) is enthusiastically explaining that creating electrical circuits and setting various parameters during the lesson with the help of the Elektro Kit has never been easier, one of the Chemistry teachers, Ms. Turkiye noted that the interactive 3D scene collection makes a real difference as it helps students thoroughly understand topics like atoms or atomic bonds.Although mozaBook provides a wide range of possibilities and content for higher grades, Ms. Taoum emphasized the importance and perks of having the interactive board and colorful pencil set on primary and kindergarten levels:
"Using different pencils or our own hands to draw shapes and write letters on the board is a great way to improve the fine motor skills of the pupils''
Interested to learn more about their journey with the mozaLearn system in the school? Check out the full video and meet the teaching crew of St. Elie here!
In case you'd like to download the mozaBook software and give it a try, just click here for the free 30-day trial version.
See you next time!


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Guidance and Inspiration for Daily Life
"Giving love to all, feeling the love of God, seeing His presence in everyone, and having but one desire — for His constant presence in the temple of your consciousness — that is the way to live in this world."
— Paramahansa Yogananda
Each of the topics below will bring you short but powerfully inspiring thoughts from the talks and writings of Paramahansa Yogananda and his close disciples. Many of the most frequently asked general questions about spiritual living are addressed in this section. Along with practical counsel about each subject, be sure to take advantage of the affirmations provided in each section. You can practice these right now to begin your personal journey toward experiencing happiness, healing, peace, and the other spiritual benefits discussed in each section.In addition, each topic includes links to resources for more in-depth exploration as well as helpful suggestions for where to find more on that subject in the many books, recordings, and other items published by Self-Realization Fellowship.


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Forum: Addons
MULTI TouchScreen
Topic: MULTI TouchScreen - Page: 1
Easy navigation.
2 Basic Views (DECKS-BROWSER)
1366x768 & 1920x12080
Version 1.1
- Fixed bug (innacurate display of FX sliders in FX panel)
- Video Mixer now has different view with Video Output
- Scratchwave on decks (toggles with HotCues - Useful when video panel is o)
- Volume on decks (toggles with Pitch - useful when Video panel is on)
- Mix_now button added
- BPM value moved to deck textzones
Version 1.2
- Added Video/Audio Crossfader button in VIdeo Mixer
- Added Nudge mode (to nudge the song instead of scratch)
- Quick loops can now act like Loop Rolls (enable the Loop Roll button)
- Added beat bars in Mixer
- Support for the SBDJ OSK Plugin http://www.virtualdj.com/addons/15940/osk.html
Version 1.3
- Fixed cue marker on songpos, not visible in Automix mode
- Fixed Set Gain to 0 when click on the Gain value
- Added 1920x1080 resolution (native HD graphics)
Note !
You can resize it to other 16:9 resolutions, but will not work with different ratios.
Enjoy !!
Inviato Sun 20 Jul 14 @ 6:00 pm
Inviato Fri 14 Nov 14 @ 8:28 pm
ald when selecting eq fx or loop ... and other side commands thers allways the fx showing goshted by the new selection i.e eq
Inviato Mon 22 Jun 15 @ 2:16 pm
Inviato Sat 25 Jul 15 @ 12:33 am
Time to have fun.. :)
Major update to the Multi-Touch Skin. Brand new graphics, 4x2 PADs matrix, Audio & Video support, new custom Browser etc.
Download link : http://www.virtualdj.com/plugins/index.html?addonid=15772
Some hints
- Tap to Artist-title to show/hide Cover on Deck (there is also an option to show cover on jog)
- Tap to Pitch text to select next pitch range
- Tap the CUSTOM pad mode twice to show custom buttons instead of the Scratchwave
- 2 Mixer modes (audio & video with the same Top panel button)
- Dual VIDEO /VIDEO FX panel
- Tap on Gain values to reset Gains to zero db
- Sync is on the BPM value button (in case you wonder :) )
The skin doesnt have any elements at the very bottom, so you are free to use the skin in semi-maximized mode and have the Windows taskbar visible (along with access to the Windows OSK when needed)
The skin doesnt force Touchscreen mode, so you can resize that (proportionaly), move it around when windowed etc.
This means that you need to choose if you need to see the Grid or the List view for the files, using the button you will see at the bottom right edge of the Songs List
Tip : Make sure the touchscreenmode setting is set to No (from Settings) or else you will get huge separator buttons :P
Enjoy !
Inviato Thu 06 Aug 15 @ 3:16 am
I Download this Version but in VDJ it Shows the Old Version
Inviato Tue 11 Aug 15 @ 8:11 pm
Kidding :)
fixed, re-download please
Inviato Tue 11 Aug 15 @ 8:30 pm
Inviato Tue 11 Aug 15 @ 8:56 pm
Inviato Fri 14 Aug 15 @ 10:21 am
Which Effect and which parameters are selected ?
Inviato Fri 14 Aug 15 @ 4:15 pm
Build 1949.790
Inviato Fri 14 Aug 15 @ 7:44 pm
Oh i see what you mean... lol
No, the mouse touchpad on your keyboard will not work (control anything).
the touchpad you see on the screen should work with touchscreens ;)
It should work with mouse if you update the build of the software to the latest one.
Inviato Fri 14 Aug 15 @ 7:57 pm
Inviato Fri 14 Aug 15 @ 8:46 pm
Inviato Tue 18 Aug 15 @ 10:59 am
Inviato Tue 25 Aug 15 @ 10:14 pm
cant drag and drop on my lenovo touch screen
Inviato Tue 01 Sep 15 @ 3:56 pm
If you have the MultitouchTwoFingerScroll setting to No, then you scroll with 1 finger and you need to drag the file from its cover in order to drag n drop it to a deck.
If you have the MultitouchTwoFingerScroll setting to Yes, then you scroll with 2 fingers and you can drag the file to a deck by touching that from any area.
MultitouchTwoFingerScroll is an Advanced Setting in Settings-->OPTIONS tab , so make sure the "Show advanced Options" is enabled.
Inviato Tue 01 Sep 15 @ 8:33 pm
Inviato Mon 14 Sep 15 @ 7:10 pm
Inviato Tue 15 Sep 15 @ 12:13 pm
Inviato Wed 16 Dec 15 @ 9:43 pm


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I am currently not on Facebook (although I did not delete my account; contemplating on returning soon too!) because I got tired of it. I have a number of reasons. This article that I saw kinda confirms that I am not alone in feeling this way. It lists the five types of annoying users:
- The Complainer – the one who spreads negative vibes with just about every status update.
- The Crazy Baby Lady – we know your baby is cute but there's no point sharing a picture every time he smiles or drools or poop.
- The Documentarian – the one who shares even the least mundane things happening around her and in her life.
- The Drama Queen – read: every little thing turns into a crisis!
- The Poser – enough said.


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Last edited by remember_Cedric; 07-20-10 at 09:09 AM.
What can I say? I'm still standing! No weapon against me shall prosper! I am more than a conqueror!!!
I don't care to sit by the window on an airplane. If I can't control it, why look?
Thanks for all his pictures! I've always liked him, such a charming actor with great potential.
Hi everyone I'm new around.
I just watched swordsman and fell in love with Lawrence so I was looking around the web for more info on him and found this forum
I think one of the reasons he was never big was because he had too many big star rivels in his generation. If he were in his twenties or thirties now, I believe he would be BIG! But anyhow he left an unforgettable impression on me in swordsman. I watched the swordsman because of Damian but I ended up enjoying Lawrence instead, although I still do like Damian.
If anyone has more info on him please keep this thread going!! And thanks so much for starting this thread in the first place.
Last edited by noquiter; 09-14-10 at 10:29 PM.
i really like him too!
i luv c~H~a~R~m~A~i~N~e
I love Lawrence. I love him best in The Swordsman (1997). He's very talented in playing such interchangeable roles there, really full of surprises from beginning to end. He plays the villain in the series but he really outshined the lead role for me in turns of appearance, sharp minded, and control. Found this clip from the series.
Ah finally found him. He was one of my most favorite actors from the 90's. I remember watching him in The Swordsman and he really stole the show. Damien Lau was great, but just a bit too old to play a 20 year old.
The more I read Liu Xing Hu Die Jian, I more I realize that The Swordsman was a very loosely adapted version of this novel. What a pity. The show itself had a really strong cast, but too bad the story was really whack towards the end.
I just love how you Captivate My Mind
Self reminder - Update blog more often and continue editing/writing for TOV fanfic.
Does anyone know what he is up to now? So sad he gets very small roles in TVB series.
For viewers or fans interested, Lawrence Yan Chi Keung was in the following
1990's ATV drama series online which I am glad I purchased on VCDs boxsets back then but some still searching for in original cantonese language.
1) Heroic Legend of the Yang Family - 30 episodes 碧血青天杨家将 - watch it now before its remove later on like other rare drama series.
2) Snow is Red 雪花神剑 - total 40 episodes
I like Lawrence Yan Chi Keung too. He did good job in portraying the storyline character of both the villian & good scholar-gentleman role.
In the beginning it was harder to find ATV drama series in its original cantonese language starred by Lawrence Yan Chi Keung
甄志強 even when I found it & was more disappointed it was out of print or out of stock.
But was glad to finally found it - ATV drama series - "The Swordsman 1996" 剑啸江湖
at http://www.tonghuacun.com/detail/18164.html#kan.
Hello Nan Ling,
Thanks for posting website link of Lawrence Yan Chi Keung 甄志強 but your posted link does not work.
To all interested viewers of ATV drama series & fans who might be interested of the Music Videos
starred Lawrence Yan Chi Keung 甄志強 uploaded by his fans:
1) MV - Heroic Legend of the Yang Family 碧血青天楊家將 at https://www.youtube.com/watch?v=GNsZTEgy2XM
2) 2nd MV - Heroic Legend of the Yang Family at https://www.youtube.com/watch?v=A4NQv3ju77Q
3) 3rd MV - Heroic Legend of the Yang Family at https://www.youtube.com/watch?v=C27ztTOMBs8
4) 4th MV - Heroic Legend of the Yang Family at https://www.youtube.com/watch?v=Bj0tmrv9SfA (if you want see images of the cast, you have to hit the forward button to skip further.)
I agreed with you on Lawrence Yan Chi Keung 甄志強.
Its too bad ATV & TVB dont give Lawrence Yan Chi Keung 甄志強 more opportunities in good leading leading roles. I think Lawrence is better off staying with ATV then switched to TVB.
At least at ATV, he is given more screen time either in occasionally good leading or major supporting roles while in TVB he is given very minor supporting roles.
I dont know what are the real reasons of why he is not given opportunities in good leading roles while other actors are.
For viewers or fans who might be interested related to Lawrence Yan Chi Keung 甄志強
1) Short biography -- http://wiki.d-addicts.com/Lawrence_Yan
2) Drama Series compliled Lawrence Yan Chi Keung starred ATV (more screen time in supporting roles or leading roles) & TVB (very minor supporting roles)
At https://movie.douban.com/celebrity/1...c&sortby=vote&
3) Online older pictures of Lawrence Yan Chi Keung
Here is what I found online in youtube which I am glad I purchased some VCDs boxsets of ATV dramas of 1990's many years ago since ATV was closed down & their dramas is rare to find in its original cantonese language.
Lawrence Yan Chi Keung 甄志強 starred in
1) ATV drama series - "The Good Lover, Bad Partner" 與狼共枕 - leading villian role
at https://www.youtube.com/watch?v=GDTltHWVuBg
2) 1990's ATV drama series 搶槓夫妻 - Unsure of the english title maybe called the couple at
Lawrence starred as one of the leading roles.
thanks for posting this MV.
I found other Music Videos uploaded by Lawrence Yan Chi Keung 甄志強 viewers-fans at
1) Music Video of ATV drama series "The swordsman" 剑啸江湖 at
2) Lawrence Yan Chi Keung was part of Thailand Music Concert at
Sub-themesong is from ATV Drama - Heroic Legend of the Yang Family 碧血青天杨家将
If anyone know of any website links related to ATV drama series which starred Lawrence, please feel free to share it to other viewers & fans.
I do wonder what he is up to now a days. Saw a few episodes of snow is red on youtube recently, miss his acting.
Recently saw an article that he is back to doing golf again and living in Shanghai.


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Songkhla, ThailandKirsty Turner
Songkhla has a lot to offer, whether you are interested in history and culture, appreciate stunning scenery or simply want to chill on the beach and swim in the sea. The town is endowed with ancient ruins, arts, and places of cultural importance. Songkhla is a melting pot of Thais, Chinese and Malays and charms visitors with its unique traditions, dialect, and folk entertainment.
To discover the area's history, the first stop should be The Songkhla National Museum, while the Phathammarong Museum is also a great source of local knowledge. The Bronze Mermaid Statue usually appears on postcards of Songkhla and represents the Hindu-Buddhist earth goddess Mae Thorani.
Songkhla is well known for its interesting architectural styles, which can best be seen in its temples and chedis. Some good examples are Wat Cha Thing Phra, Wat Pha Kho, Wat Chai Mongkhon and Wat Matchimawat. The city's black and white stupas - known as Chedi Ong Dam and Chedi Ong Khao - should not be missed and Sating Phra Ancient Community is well worth a visit.
Songkhla also contains some areas of stunning natural beauty. Top of the list are the Khao Nam Khang National Park with its jungle, caves and waterfalls and Khu Khut Waterfowl Park. As its name suggests, Namtok Boriphat Forestry Park features a large number of waterfalls and beautiful forest, while Wat Tham Khao Rup Chang is an interesting cave temple.
Songkhla is blessed with a large number of caves to explore and mountain tops offering spectacular views over the area. A good place to start is Khao Nam Khang Historic Tunnel, while other mountains include Khao Tang Kuan, Khao Kao Seng and Khao Noi.
There are some very pretty beaches to soak up the sun on including Hat Samila and Hat Sakom, while Hat Yai is the liveliest town and famous for fresh seafood and Muay
Thai boxing matches. Whilst in Hat Yai, pay a visit to Wat Hat Yai Nai, which features a 35 meter reclining Buddha known as Phra Phut Mahatamongkon and the very pretty and peaceful Hat Yai Municipal Park.
Amongst the area's small and somewhat secluded islands are Koh Maeo and Koh Nu (cat and mouse islands) and Koh Yo, which is a very pretty island famous for its cotton weaving community.
Of course, when it comes to eating, seafood dominates the menu. The best place to find a good selection of reasonably priced seafood is at the local night markets, where you can relax for a while at one of the small tables and watch the dramas of this charismatic area unfold around you.


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As a NEA member, when you post your request for up to $500 for classroom materials on DonorsChoose.org, the NEA Foundation will match the donations you receive, up to $250 per request. For every dollar given, we'll give one too!
Last year, we helped fund close to 1,500 NEA-member project requests, reaching more than 120,000 public school students.
This year, with your support, we hope to reach even more!
Have a great idea? Post your project requests for classroom materials.
Looking for back-to-school ideas, or want to support other teachers? Search for a project by location, subject, or age.
Don't forget to check back later when we'll be sharing some of the innovative projects ideas from DonorsChoose.org.


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Macrophage polarization in response to stimuli within the microenvironment results in heterogeneous populations that can differentially influence disease. Atherosclerosis is largely inflammatory, with lipids driving polarization of M1 macrophages; however, alternatively activated, hemoglobin-scavenging CD163+ macrophages are present within atherosclerotic lesions and have been proposed to serve an antiinflammatory role. In this episode, Aloke Finn and Liang Guo discuss their work, which shows that CD163+ macrophages actually promote angiogenesis, vessel permeability, and leucocyte infiltration, thereby exacerbating plaque progression. The pathogenic effects of this alternatively activated population were driven by HIF1α/VEGF-A signaling, suggesting this pathway as a potential therapeutic target.
Intake of hemoglobin by the hemoglobin-haptoglobin receptor CD163 leads to a distinct alternative non–foam cell antiinflammatory macrophage phenotype that was previously considered atheroprotective. Here, we reveal an unexpected but important pathogenic role for these macrophages in atherosclerosis. Using human atherosclerotic samples, cultured cells, and a mouse model of advanced atherosclerosis, we investigated the role of intraplaque hemorrhage on macrophage function with respect to angiogenesis, vascular permeability, inflammation, and plaque progression. In human atherosclerotic lesions, CD163+ macrophages were associated with plaque progression, microvascularity, and a high level of HIF1α and VEGF-A expression. We observed irregular vascular endothelial cadherin in intraplaque microvessels surrounded by CD163+ macrophages. Within these cells, activation of HIF1α via inhibition of prolyl hydroxylases promoted VEGF-mediated increases in intraplaque angiogenesis, vascular permeability, and inflammatory cell recruitment. CD163+ macrophages increased intraplaque endothelial VCAM expression and plaque inflammation. Subjects with homozygous minor alleles of the SNP rs7136716 had elevated microvessel density, increased expression of CD163 in ruptured coronary plaques, and a higher risk of myocardial infarction and coronary heart disease in population cohorts. Thus, our findings highlight a nonlipid-driven mechanism by which alternative macrophages promote plaque angiogenesis, leakiness, inflammation, and progression via the CD163/HIF1α/VEGF-A pathway.
Liang Guo, Hirokuni Akahori, Emanuel Harari, Samantha L. Smith, Rohini Polavarapu, Vinit Karmali, Fumiyuki Otsuka, Rachel L. Gannon, Ryan E. Braumann, Megan H. Dickinson, Anuj Gupta, Audrey L. Jenkins, Michael J. Lipinski, Johoon Kim, Peter Chhour, Paul S. de Vries, Hiroyuki Jinnouchi, Robert Kutys, Hiroyoshi Mori, Matthew D. Kutyna, Sho Torii, Atsushi Sakamoto, Cheol Ung Choi, Qi Cheng, Megan L. Grove, Mariem A. Sawan, Yin Zhang, Yihai Cao, Frank D. Kolodgie, David P. Cormode, Dan E. Arking, Eric Boerwinkle, Alanna C. Morrison, Jeanette Erdmann, Nona Sotoodehnia, Renu Virmani, Aloke V. Finn


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Welcome to My Personal Page
Tracy Murillo
Goal: $200.00
Achieved: $85.00
Achieved: $85.00
Fundraising Honor Roll
Tracy Murillo
Please support me as I take a stand behind the brave men and women of law enforcement who put their lives on the line every day to protect the public.
I'm a part of the Run for the Badge because law enforcement officers support us every day, and every 60 hours an officer makes the ultimate sacrifice for public safety. I'm showing my support by participating and I hope you'll help me by making a generous donation toward my fundraising goal.


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I want to export an environment variable whose value contains a variable but I don't want that variable (which is within the value) to be evaluated during the export process. i.e.
export STY=for i in {0..3}; do echo $i; done
so my requirement is that when I echo STY, the output that I need should be
echo $STY
for i in {0..3}; do echo $i; done
But $i is evaluated during export. I need to preserve it as it is.


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Today is the death-day of St. Catherine (1380). She was 33. She practiced an extreme asceticism, beginning when she was 7, and joined the Dominicans as an anchorite, living in a hermitage, when she was 16. She is remembered both for her mystical experiences and her practical diplomacy between popes, kings and individuals alike. She wrote extensively and we still have her Treatise on Divine Providence, which is considered the mystical counterpart to Dante's Divine Comedy; a collection of 400 letters; and her prayers. You can read more about her here.
Here is part of one of her most famous prayers:
"O unfathomable depth, o deity eternal, o deep ocean, what more could you give me than to give me yourself? You are an ever-burning fire. You consume and are not consumed. By your fire, you consume every trace of self-love in the soul. You are a fire which drives away all coldness and illumines minds with its light, and with this light you have made known your truth. Truly this light is a sea which feeds the soul until it is all immersed in you, o peaceful sea, eternal Trinity! The water of this sea is never turbid. It never causes fear, but gives knowledge of the truth. This water is transparent and discloses hidden things. And a living faith gives such abundance of light that the soul almost attains to certitude in what it believes."
St. Catherine was a peace-maker. Peace is both an outward fruit of the Spirit (Gal 5:22–23) and an inward longing that demonstrates God's presence to us. Let's do peace in our haikus and images today.
See Prayer Light.


text: Cody Regional Health Rheumatology Clinic Comprehensive Treatment for Autoimmune Diseases To schedule an appointment with our Internal Medicine Specialty Clinic, please call 307-578-2975. Rheumatoid diseases are conditions where the body's own immune system turns on itself and starts attacking healthy tissue. These diseases typically manifest as joint and muscle pains, like arthritis. We still do not know what causes these conditions or how to cure them, but there are numerous treatments available to manage the symptoms. Why might you need to see a Rheumatologist? Here are some common disorders treated: Gout and pseudogout Osteoarthritis Polymyalgia rheumatica Psoriatic arthritis Raynaud's disease Rheumatoid arthritis Seronegative inflammatory arthritis Systemic lupus erythematosus (lupus) Spondylitis disorders (ankylosing spondylitis and related conditions) Cody Regional Health's Rheumatology Clinic offers state-of-the-art diagnosis and treatment of rheumatic diseases in adults. Our providers have been trained in Internal Medicine and Rheumatology and they work closely with specialists in other departments to provide our patients with coordinated and comprehensive care. Treatment Strategies Treatment for Rheumatoid diseases focuses on controlling and mitigating the symptoms. Arthritis is a common problem, and one medical professionals have been treating for decades. Today, treatment has advanced to the point where we can provide long-term treatment that is minimally invasive and highly effective. Below are a few of the many services we offer in our clinic: Outpatient Rheumatology consultations Collaborative evaluations incorporating doctors from other specialties Access to state-of-the-art laboratory, imaging and pathology services A shared electronic medical record, that contains all consultation reports, laboratory results, radiology and procedure reports Outpatient specialty care for patients with chronic rheumatic diseases Hospital consultations Injections Our Rheumatologists will work closely with you to better understand the challenges you face on a daily basis as a result of your condition. By learning more about you, we can create more effective, personalized treatment plans to help you enjoy a higher quality of life. Mission To provide extraordinary healthcare to those we serve by people who care. Vision The first choice for those who aspire to heal, be healed and lead healthier lives. Key Values Compassion, Integrity, Excellence, Innovation and Collaboration. 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It's all too rare that an artist appears, seemingly out of nowhere, fully matured and with a distinctively personal musical conception. While guitarist Nguyęn Lę—Parisian born of Vietnamese parents—might argue that his true voice didn't emerge until he released the seminal Tales From Vietnam (ACT, 1996), the recent remastered reissue of his first two albums as a leader, Miracles and Zanzibar, suggests that Lę may be too hard on himself.
It's also not necessarily true that you are defined by the company you keep. Plenty of young artists—especially if they're backed by a label with some cash—hire known names for their first recording as a leader. Still, while these big guns are rarely less than accomplished, it's easy enough to tell when they're simply "doing the session" as opposed to finding themselves deeply committed to the music. And it's rarer still to find better-known musicians maintaining associations with a younger artist once the session is over.
What's most remarkable about Lę's first two recordings—one representing a literal first meeting, the other demonstrating the benefit of having been on tour prior to recording—is not just how cohesive the two groups are, but that they represent the beginnings of musical friendships that continue to this day. Pianist Art Lande, who is on both sessions, recently rejoined Lę on Walking On The Tiger's Tail (ACT, 2005). That release also featured Oregon co-founder and woodwind multi-instrumentalist, Paul McCandless, who also appears on one of Lę's early sessions. Bassist Marc Johnson and drummer Peter Erskine, from Lę's first release, would work again with Lę on Three Trios (ACT, 1997), while Erskine would continue the relationship, forming the collaborative E_L_B trio with Lę and releasing its self-titled ACT album in 2001.
That so many of these artists clearly heard something in Lę at such an early stage makes these reissues long overdue.
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The Little Johnny Range Digital Cottons Rainbow Butterfly
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Atmospher Sci & Global Chg
Research Highlights
December 2013
Building a Structure for Efficiency
Research is laying the foundation for improving energy use in rural China
Low efficiency heating equipment, such as this Kang wood stove, illustrates the challenge for researchers and building standards officials in rural China. As part of a whole-house heating unit, this stove and other low-efficiency heating and cooking units can be replaced with high-efficiency models as a part of new building energy codes implementation. Photo courtesy of CABR. Enlarge Image.
Results: Researchers at Pacific Northwest National Laboratory, working with partners at the China Academy of Building Research (CABR), analyzed current and future building energy use in rural China and developed an assessment of the issues and options for improving capacity and implementation of a building energy standard. Though there are feasible options for improving energy efficiency in rural buildings, they researched the significant challenges in establishing support at a local level. Due in part to their work, China has recently adopted a voluntary building energy standard to guide local governments in their work with rural populations. Their research is helping China to take important steps to improve energy efficiency in rural buildings.
"The Chinese government was interested in testing the feasibility of a code before adopting it," said Meredydd Evans, lead researcher and expert in building energy efficiency working at the Joint Global Change Research Institute (JGCRI) in Maryland. "The pilot implementation projects and reviews we conducted with CABR helped establish the feasibility of moving forward with a code on a voluntary basis."
JGCRI is a partnership between PNNL and the University of Maryland.
Why It Matters: Worldwide energy use demands are increasing, and so are concerns to limit greenhouse gas emissions and climate change. China, with one fifth of the world's population, has a growing economy and per capita energy use. Rural buildings now account for well over half of China's total building energy use and approximately 40 percent of their total floor space, equivalent to all U.S. residential floor space. As rural families switch to commercial fuels and expand their use of energy, the emissions from these buildings will grow. Because of their current inefficiency, improving energy efficiency in rural buildings is a readily achievable goal—a "low hanging fruit"—for capturing energy savings with feasible options. The PNNL researchers, collaborating with partners in China, are tackling the issues of how to improve energy efficiency and building standards in rural China.
Replacing inefficient home equipment with efficient models, such as these cooking and heating appliances, will reduce energy use and improve home air quality as well. Photo courtesy of CABR. Enlarge Image.
Methods: A PNNL research team led by Evans working at JGCRI used the Global Change Assessment Model (GCAM) to assess China's rural building energy trends. GCAM, developed at PNNL with support from the U.S. Department of Energy, is an integrated assessment model that links economic, energy, land-use and climate systems to show the effects of regional and global change. The team conducted field interviews with Chinese experts to understand the challenges and options for rural building codes to promote efficient energy use. They developed recommendations on possible policy options and implementation mechanisms for new codes, and tested implementation options for a rural building code in two rural villages in conjunction with the CABR.
What's Next? China adopted a voluntary rural building energy standard in December 2012. Now researchers are moving to assess progress with implementation, and as capacity grows, move toward making the standard a requirement.
Sponsor: The U.S. Department of Energy Office of Energy Efficiency and Renewable Energy and the Global Technology Strategy Program.
Research Team: Meredydd Evans, Sha Yu and Alison Delgado, Pacific Northwest National Laboratory; Bo Song, Qinqin Deng and Jing Liu, China Academy of Building Research, China.
Reference: Evans M, S Yu, B Song, Q Deng, J Liu, and A Delgado. 2014. "Building Energy Efficiency in Rural China." Energy Policy 64:243-251. DOI:10.1016/j.enpol.2013.06.040


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Collingwood, VIC – Jims Tree and Stump Removal, a company specialising in trees, provides services pertaining to tree removals, stump grinding, cleaning services and more. All services come at affordable costs and are performed by tree experts, ensuring quality work. This company has varied services available, helping customers in more than one way.
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David Brennan
Born and raised in New Orleans, David Brennan attended the University of New Orleans where he received a Bachelor of Arts degree in Political science. He was a Louisiana District manager for the 1984 re-election campaign of Ronald Reagan. Since 1992 he has studied Bible prophecy as it relates to current international political issues.


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Thank you for visiting Changing Channels. This blog is no longer being updated. Please enjoy the archives. Here is another blog you may also enjoy: Idol Chatter
Kirstie Alley will have a heckuva time winning "Dancing With the Stars" this season.
The other dancers are younger, more talented and yes, thinner than the former "Cheers" star.
There's still something pretty cool about Alley hanging in there show after show and doing the best she can to compete.
The other competitors are more fluid and easier to watch, but Alley is the most "American" of any of them.
Meaning her on and off weight issues, similar to Oprah and like so many Americans makes her more like many of "us".
While I cringed at Alley at the start of the season, particularly after she fell, I have re-learned an important lesson from watching her persevere week after week.
Everyone falls at one time or another … even "stars" that people look up to!
So way to go Kirstie … you did fall. But you certainly got back up and have become an inspiration along the way!


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Certificate Courses for Academic LeadersCopyright: Thilo Vogel
Program Coordinator
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- Send Email
Academic leaders have the opportunity to obtain the "Leadership" certificate and, if desired, complete the supplementary "Research and Teaching" module. The Leadership certificate is to testify your competencies in leadership and management. If you are active in research and teaching, you can also complete the complementary module. Your participation will be noted on the Leadership certificate.
There are two ways to acquire the "Leadership" certificate:
1. For the "Leadership" certificate, participants can freely select seminars from four modules leadership, personal development, motivation and commitment, and we-feeling:
- Module Leadership – at least four seminar days; participation in the three-day workshop "Leadership at RWTH" is mandatory
- Module Personal Development – at least three seminar days
- Module Motivation and Commitment – at least two seminar days
- Module "We-Feeling" – at least one seminar day
If you are interested in participating, please register for the course by writing an e-mail to Dagmar Grübler. Registered participants in the certificate course will be specifically invited to the three-day workshop "Leadership at RWTH."
2. You complete the "Qualification Course for Leaders in Science, Adminstration, and Technology." The target group of people in science for this course are tenured employees at the academic mid-level. The qualification course is held during six two-day events with a set group of participants. The course lasts about one year. To register, please send an email to Dagmar Grübler.
As a complement to the Leadership course, you can also obtain the "Teaching and Research" certificate. This requires prior participation in workshops from the modules "Research and Innovation Capability" and "Teaching Competence". Participants can freely select workshops from these modules.
- Module Research and Innovation Capability – at least three seminar days
- Module Teaching Competence – at least one seminar day
For an overview of workshops and seminars and their module categorization, please refer to our website, which also provides individual seminar descriptions. To obtain your certificate, please present the relevant certificates of attendance to Ms. Dagmar Grübler. Participation certificates from other RWTH continuing education providers are also accepted.Copyright: private
I was delighted with the leadership course! The group was a good mix of researchers and administrative staff; the coach was confident, knowledgeable and process-focused; and the collaboration with my colleagues was fun and inspiring. The content and methodical approach help me a lot in my role as a leader. I am already looking forward to the next modules of the course and would like to say thank you for this opportunity to learn and develop!
Dr.-Ing. Bernhard Schmenk, Head of Corporate Development and Communications, ITA, RWTH Aachen University