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A novel method is provided for purifying DNA and RNA from a variety of sources, including cells, cell lysates, viruses, tissues, blood and other body fluids employing a cationic detergent to complex with the nucleic acids.
1. A process for the purification of DNA and RNA from a biological mixture consisting essentially of applying directly to said mixture without prior purification or extraction of the nucleic acids a selected quaternary amine cationic detergent capable of simultaneously solubilizing contaminants in said mixture and forming a hydrophobic complex with said DNA or RNA in said mixture.
2. A proceeds for the purification of DNA and RNA from a biological mixture containing cells, lysed or intact, consisting essentially of applying directly to said mixture without prior purification or extraction of the nucleic acids a selected quaternary amine cationic detergent capable of simultaneously solubilizing contaminants in said mixture and forming a hydrophobic complex with said DNA or RNA in said mixture.
3. A process for the purification of DNA and RNA from a biological mixture containing virus consisting essentially of applying directly to said mixture without prior purification or extraction of the nucleic acids of selected quaternary amine cationic detergent capable of simultaneously solubilizing contaminants in said mixture and forming a hydrophobic complex with said DNA or RNA in said mixture.
4. The process according to claim 1 further consisting essentially of separating said hydrophobic complex from said contaminants.
5. The process according to claim 4 wherein said separating step consists essentially of centrifuging said mixture following the addition of fine particles of matter for facilitating precipitation of said complex.
6. The process according to claim 4 wherein said separating step consists essentially of collecting said complex on a surface or a matrix by filtration or absorption.
7. The process according to claim 1 further consisting essentially of dissociating said DNA and RNA from said complex to produce purified DNA and RNA.
8. The process according to claim 7 wherein said dissociating step comprises treating said complex with a salt in a non-aqueous solvent.
9. The process according to claim 1 wherein said detergent is an alkylbenzyldimethylammonium salt.
10. The process according to claim 9 wherein said detergent is an alkylbenzyldimethylammonium chloride, wherein said alkyl group consists essentially of from 10 to 20 carbon atoms.
11. The process according to claim 10 wherein said alkyl group consists essentially of 16 carbon atoms.
12. The process according to claim 1 further consisting essentially of adding to said mixture an chaotropic agent.
13. The process according to claim 1 further consisting essentially of adding to said mixture an sulfhydryl-reducing agent.
14. The process according to claim 1 further consisting essentially of adding to said mixture an chelating agent.
15. The process according to claim 1 further consisting essentially of adding to said mixture an non-cationic detergent.
16. The process according to claim 10 wherein said alkyl group consists essentially of 14 carbon atoms.
17. The process according to claim 10 wherein said alkyl group consists essentially of 18 carbon atoms.
18. The process according to claim 1 wherein said biological material consists essentially intact cells, cell lysates, blood, viruses, or other body fluids or solutions containing DNA or RNA.
19. A kit for isolating nucleic acids from a sample biological mixture containing nucleic acids in contaminating material consisting essentially of a cationic detergent in an amount sufficient to solubilize contaminants in said sample and form a hydrophobic complex with nucleic acids present in said sample.
20. The kit according to claim 19 wherein said detergent using a quaternary amine cationic detergent.
21. The kit according to claim 19 wherein said detergent is an alkylbenzyldimethylammonium salt.
22. The kit according to claim 21 wherein said detergent is an alkyl benzyldimethylammonium chloride.
23. The kit according to claim 22 wherein said alkyl group consists essentially of from 10 to 20 carbon atoms.
24. The kit according to claim 23 wherein said alkyl group consists essentially of 16 carbon atoms.
Deoxyribonucleic acid (DNA), and ribonucleic acid (RNA), polymers of nucleic acids arranged in a particular order which contain inheritable genetic information, are employed in a wide variety of research, medical, diagnostic and industrial processes. The variety of uses for extracted and purified DNA and RNA from disparate sources is rapidly increasing with the advent of biotechnology as an industry for the production of pharmaceutical, agricultural, pesticidal and other agents. Additionally nucleic acid sequences are being increasingly employed for their ability to detect and identify genetic and familial disorders and carrier states; genetic aberrations found in tumors; and proof of identity or parentage. Nucleic acid sequences are also being employed to aid in the detection of infections by bacteria, viruses and other agents.
The production of genetically engineered proteins and polypeptides is another area where purified nucleic acid sequences are in demand. DNA and RNA are employed as starting materials in the manufacture of a variety of products, including nucleoside antibiotic and antiviral agents. DNA and RNA libraries and clones selected from them are also routinely employed in molecular biology and biotechnological research.
Generally nucleic acid sequences must be extracted from biological sources, e.g., tissue samples, bacteria, viruses, salmon sperm and the like, and purified by separation from proteins, salts and other biological molecules prior to use.
The standard procedure for isolating DNA from a cell source involves digestion with a combination of a proteolytic enzyme and a non-ionic or anionic detergent, such as sarcosyl or sodium dodecyl sulfate. The resulting digest is extracted with a mixture of phenol and chloroform, which removes most of the hydrolyzed products. The DNA is then precipitated from the resulting aqueous phase by the addition of alcohol.
For example, genomic DNA is extracted from eukaryotic cells by incubating them with a proteolytic enzyme (usually proteinase k) and the anionic detergent, SDS (sodium dodecyl sulfate). The resulting mixture is extracted with a mixture of phenol and chloroform, which leaves the DNA and RNA in the aqueous phase. The DNA is then precipitated by the addition of ethanol and sodium acetate, and resuspended in a buffer. RNase is then used to hydrolyse the RNA, and the DNA is collected by phenol/chloroform extraction and ethanol precipitation. Alternatively, cell nuclei can be prepared by dounce homogenization (or frozen pulverisation), filtration and centrifugation through sucrose. The nuclei are treated with proteinase K and SDS as above. Preparations of DNA should not contain impurities which inhibit the enzymes used to manipulate it further, such as restriction endonucleases or the Tag polymerase for use in the polymerase chain reaction. For some applications, the DNA should be very long. In these cases, DNA is prepared by the same general methods, but applied to cells imbedded in an agarose gel.
DNA in plasmids exists as double-stranded, closed circular DNA in the host bacteria. It is harvested by lysing the bacteria with lysozyme and a nonionic detergent. This liberates the plasmids, but leaves most of the bacterial DNA adherent to the cell debris, which can be removed by ultra-centrifugation. The plasmid is then separated by density-gradient ultra-centrifugation in the presence of ethidium bromide. Alternatively, the bacteria are treated with sodium hydroxide, and the plasmid DNA is separated by exclusion gel chromatography after RNase treatment.
DNA from bacteriophages or other viruses can be collected (after removing contaminating bacterial DNA with a DNase) by precipitation with polethylene glycol followed by extraction with chloroform/phenol and ethanol precipitation.
DNA is usually purified from reaction mixtures by chloroform/phenol extraction and ethanol precipitation. Spermine can be used to precipitate DNA, and can subsequently be removed by dialysis. DNA can be harvested by its binding to powdered glass in the presence of high salt solutions, or by adsorption and elution from commercially prepared columns.
The extraction of full length RNA requires methods that inhibit RNase's, which are ubiquitous. Cells can be ruptured by the addition of SDS in the presence of an RNase inhibitor, and extracted with phenol/chloroform followed by ethanol precipitation. Alternatively, cells can be lysed by the addition of guanidinium isothiocyanate followed by ultra-centrifugation through a cesium chloride gradient.
RNA is extracted by a variety of techniques designed to protect it from the action of RNAase's. One conventional method employs the chaptropic agent, guanidinium isothiocyanate, followed by ultracentrifugation to harvest the dense RNA. Another standard method for obtaining RNA from cell sources uses hot phenol extraction, followed by digestion of DNA with DNAase, extraction with phenol/chloroform, and precipitation with alcohol. In the intermediate steps of the processing of DNA, phenol/chloroform extraction and ethanol precipitation are often used.
The above-described nucleic acid extraction and purification techniques are described in detail in a number of standard molecular biology methodological texts, including T. Maniatis et al., "Molecular Cloning. A Laboratory Manual.", Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y. (1982) and L. G. Davis et al, "Basic Methods in Molecular Biology", Elsevier, N.Y. (1986).
A number of cationic detergents have been shown to be able to precipitate DNA and RNA from aqueous phases. Cationic detergents consist of a positively charged head group, which is usually a quaternary amine or a pyridinium group, and an aliphatic tail. Examples of commercially useful detergents include cetyl pyridinium bromide, cetyltrimethylammonium bromide (collectively known as cetrimonium compounds) and alkylbenzyldimethylammonium chlorides (collectively known as benzalkonium compounds). As the length of the side chain is increased, the resulting detergent become stronger, and its solubility in water decreases. Typical cationic detergents used in these procedures are cetyl pyridinium bromide, and cetyl trimethylammonium bromide, among others. See, e.g., A. S. Jones, Nature, 199:280-82 (1963); J. H. Weil and J. P. Ebel, Biochem. Biophys. Acta., 55:836-840 (1962). The ability of cationic detergents to precipitate DNA and RNA was reported by A. S. Jones, Biochem. Biophys. Acta.. 10:607 (1953). In a typical application, micro-organisms were extracted with phenol-p-aminosalicylate, and the DNA in the extract was precipitated with ethanol. The precipitate was dissolved in IM NaCl (in which cytoplasmic RNA is insoluble), and the DNA was precipitated by the addition of cetyl trimethylammonium bromide (CTAB), after dilution to 0.55 to 0.60M NaCl. The DNA/CTAB complex was redissolved in 1M NaCl and reprecipitated with ethanol. The solubility of RNA and DNA complexes with cationic detergents in polar organic solvents was reported by French researchers. These cationic detergents included cetrimonium compounds and benzalkonium bromides, and the solvents included ethanol and formamide. These workers showed that the nucleotides could be precipitated from the organic solvent by the addition of sodium chloride. The ability of benzalkonium compounds to solubilise proteins at the same time as precipitating nucleotides was not mentioned. Other workers have reported similar findings.
There remains a need in the art for additional, simpler and more efficient methods for extraction and purifying nucleic acids from cell sources.
In one aspect the invention provides a method for purifying DNA or RNA from a mixture of biological materials, which comprises the step of adding a cationic detergent to a mixture containing the RNA or DNA in an amount sufficient to dissolve cells, solubilize any contaminating proteins and lipids in the mixture, and form insoluble hydrophobic complex between the nucleic acid and the detergent. The complex which comprises the RNA or DNA with the detergent is separated from the solubilized contaminants, and may be dissolved or dispersed in a polar organic solvent. Thereafter the DNA or RNA is recovered by the addition of a salt, which promotes the dissociation of the complex.
As another aspect of the present invention, there is provided a kit for use in performing the method of this invention comprising a quaternary amine cationic detergent. Additional optional components of the kit are one or more chaotropic, sulfhydryl-reducing or chelating agents, non-cationic detergents or hydrolytic enzymes.
Other aspects and advantages of the present invention are described further in the following detailed description.
The present invention provides a novel method for purifying RNA and DNA from a mixture of biological materials. The biological material mixture may be intact cells. Alternatively, the mixture may be cell lysates, blood, viruses, other body fluids or other solutions containing DNA or RNA plus other contaminating molecules. This method is advantageously characterized by greater simplicity, speed and economy than prior art methods of nucleic acid purification. Additionally the process of this invention is less hazardous and more amenable to scale up or miniaturization than currently available methods.
The method of the present invention employs the step of treating a mixture of biological materials containing DNA or RNA with a cationic detergent to solubilize the contaminating proteins and lipids therein and cause the formation of a hydrophobic complex between the nucleic acids and the detergent.
Surprisingly, although some cationic detergents, e.g., cetyl pyridinium bromide, have been employed to precipitate DNA or RNA from aqueous media, the use of cationic detergents to directly treat the biological sources of nucleic acid to lyse cells and simultaneously precipitate RNA and DNA therefrom has not been described. Applicant has surprisingly discovered that a simpler, and more direct method of nucleic acid purification results from the use of a cationic detergent as a solubilizing agent.
According to the method of the invention, a cationic detergent is added to a mixture of biological materials containing nucleic acids. In practice of this invention to the biological mixture is added an aqueous solution in an amount sufficient to provide a final mixture comprising approximately 0.1 to 10% of the selected cationic detergent and from about 1 to 1000 μg nucleotide per ml. The detergent lyses the cells, and solubilizes contaminating lipids and proteins present in the mixture. The DNA and/or RNA in the mixture comprising the biological fluid and detergent solution form a hydrophobic complex with the detergent. This hydrophobic complex is then separated easily from the solubilized contaminants.
The cationic detergent selected for use in the method of the present invention is characterized by solubility in, or miscibility with, the extracting mixture. It must be a powerful detergent capable of lysing cells and solubilizing proteins and lipids. It is also capable of forming a hydrophobic complex with DNA or RNA, sufficiently strong to remain intact during the separation step. Further, the cationic detergent must not have deleterious effects, or contain impurities with such effects, on the subsequent processing of the RNA or DNA.
Presently preferred cationic detergents for use in the present invention are therefore quaternary amine detergents. Preferably, the detergent is an alkylbenzyldimethylammonium salt. More preferably, the detergent is an alkylbenzyldimethylammonium chloride, particularly where the alkyl group is linear and contains between 10 and 20 carbon atoms. The presently most preferred detergent is an alkyl-benzyl dimethyl-ammonium chloride, wherein the alkyl group contains 16 carbon atoms in a straight chain. This detergent, which is referred to hereafter as "16-BAC", can be prepared conventionally as a concentrated solution in water by techniques known to those of skill in the art. This detergent, also called benzyl dimethyl hexadecyl ammonium chloride, may be commercially obtained from Sigma Chemical Co., St. Louis, Mo. However other cationic detergents which fulfill the above-stated requirements may be selected for use in the practice of the invention by one of skill in the art.
The method of this invention may optionally include other steps such as adding to the mixture of biological materials, singly or in combination, one or more chaotropic, sulfhydryl-reducing or chelating agents, non-cationic detergents or hydrolytic enzymes. These optional components of the present invention may be added to the mixture before, during or after the addition of the cationic detergent. Alternatively, these optional components may be mixed with the cationic detergent prior to adding the detergent to the biological material.
The efficiency of the solubilization of contaminating substances may be enhanced in the method of this invention by the optional addition with the cationic detergent of a chaotropic agent to a final concentration in the biological mixture of about 0 to 60%. One typically employed agent is urea.
Another optional additive to the mixture undergoing the method of this invention is a sulfhydryl reducing agent, such as 2-mercaptoethanol or other known agents. A sulfhydryl reducing agent may be added to the mixture of biological materials to provide in a final concentration of about 0 to 2%.
A chelating agent, such as ethylenediaminetetraacetic acid (EDTA), may also be optionally used with the cationic detergent in this method to provide a concentration of from 0 to 2% in the final mixture.
The nucleic acid yield may be further increased by the addition to the mixture of hydrolytic enzymes, such as proteinase K, resulting in a final concentration of from 0 to 1% and/or non-cationic detergents, such as, Triton X100, resulting in a final concentration of from 0 to 10%, to disperse tissues before or during the addition of the cationic detergent.
According to the method, a hydrophobic complex of RNA or DNA with the detergent results upon treating the biological mixture with the detergent. This complex may be collected by centrifugation employing a force adequate to sediment the precipitate but not so large as to compress it into an intractable pellet. Such a centrifugation force increases as the content of the nucleotide decreases, and may desirably range from 100 xG to 15,000 xG for approximately 1 to 20 minutes. Precipitation may be enhanced by the addition of fine particles, such as silica gel.
Alternatively, the hydrophobic nucleic acid-cationic detergent complex may be separated from the solubilized contaminants by filtration. A conventional filter or matrix may be selected to capture the complexes without the use of excessive pressures. Illustrative filters for such use desirably have large pore sizes and may include Whatmann GF-A glass paper. The following standard matrices may also be employed in this separation step: gel filtration beads, hydrophobic beads, ion exchange beads, or hydrophilic beads.
The complexes can be washed with aqueous solutions to remove any residual contaminating substances. The hydrophobic nucleic acid-cationic detergent complexes are then dissociated by treatment with a suitable cation to liberate the DNA and RNA as their respective salts. This step involves dissolving or dispersing the complexes in a non-aqueous solvent, and then adding a suitable salt, which precipitates the RNA and DNA from the solvent, leaving the detergent in solution. Alternatively, the hydrophobic complexes may be dissociated and the RNA and DNA recovered by the addition to the complex of the salt without the prior dispersion or solution of the complexes in the solvent.
Many salts are capable of use in this dissociation step. Presently preferred salts are sodium chloride, sodium acetate, ammonium acetate. Although many solvents are suitable for this purpose, presently preferred solvents for use in this step of the method include ethanol, methanol and formamide and mixtures thereof. The dissociation steps takes approximately 5-120 minutes, with gentle stirring or agitation.
The method of this invention may be preferably performed, particularly for diagnostic purposes, such as the isolation of nucleic acids from a tumor biopsy sample, and other commercial purposes, by use of a kit providing the following minimal reagents required for the method. Such a kit includes specifically approximately 1 ml of solution of a selected cationic detergent, e.g., 16BAC, for use in treatment of approximately 10 mg of a tissue sample, solution, and the like. Approximately 10 ml each of a suitable solvent and a suitable salt solution for the dissolution step of the method are also included in the kit. Also part of such a kit are approximately 1 mg of an optional proteolytic enzyme, e.g., proteinase K for mixing with the biological fluid; an optional 0.1 ml of an anticoagulant for processing blood samples, and an optional 1 ml of a nonionic detergent solution for lysing cells. The kit may also comprise a container for mixing the biological fluid or tissue with the detergent and a suitable filter or matrix for the separation step. Other conventional components for diagnostic applications may also be included in a kit for performance of the method of the present invention.
The following examples illustrate performance of the method of the present invention, and are not intended to limit the scope of this disclosure.
Human leukemic blood cells (15 million HL-60 cells) were employed as the DNA source in this example. The cells were washed with Hanks solution and centrifuged to a pellet. To this pellet was added 5 ml of an aqueous solution containing 1% 16-BAC, 1% 2-mercaptoethanol, 10 mM EDTA, 4 M urea, and the resulting suspension was gently shaken. After standing for 15 minutes at room temperature, this mixture was centrifuged at 800 g for 10 minutes. The supernatant was discarded, and the pellet was washed by dispersing it in water and centrifuging it twice. The pellet was then dissolved in 0.7 ml ice cold ethanol by gentle agitation for 2 hours. Seven-tenths ml of ethanol saturated with ammonium acetate was then added, and the DNA and RNA was allowed to precipitate at -20� C. for 2 hours.
The precipitate was dissolved in 0.5 ml 10 mM tris HCl pH 7.5, 15 mM NaCl, treated with RNAase and EcoR1 enzymes by conventional techniques. The yield of DNA was estimated by ultraviolet spectroscopy (OD260 ) as 28, 47, 61, and 42 micrograms. The OD260 /OD280 ratios were between 1.56 and 1.71. After electrophoresis through agarose, blotting to a filter and hybridization with a radioactive c-myc probe, the expected EcoRI-digested restriction fragment of the myc gene of 12.8 kb was detected by autoradiography.
HL-60 cells (15 million) were washed by centrifugation and resuspension in 5 ml of phosphate buffered saline. Five ml of an aqueous solution containing 2% 16-BAC, 40% urea, 2% 2-mercaptoethanol, and 20 mM EDTA was added to the suspension. The resulting mixture was centrifuged at 600 g for 5 minutes. The pellet was washed twice by resuspension in water and centrifugation, and then the pellet was extracted with 1 ml ethanol by vortexing for a few minutes. The mixture was centrifuged, and the supernatant was collected.
Fifty microliters of 3 M aqueous sodium acetate pH 5.2 and 400 microliters of water were added and the nucleotides were allowed to precipitate at -20� C. overnight. After centrifugation, the pellet was dissolved in 20 microliters of RNAase free water. The yield of nucleotide (by OD260) was 67 micrograms, OD260 /OD280 =1.59. Denaturing agarose gel electrophoresis (using formaldehyde) followed by elhidium bromide staining revealed fluorescent bands expected for undegraded rRNA. Northern blotting and hybridizing with a c-myc probe revealed c-myc mRNA.
E. coli (2.24�109 cells in 4 ml culture medium) were added to the 16-BAC extraction mixture of Example 2, and extracted following identical procedures. The yield of nucleotide, measured by OD260, was 261 micrograms, OD260 /OD280 =1.92. Denaturing agarose gel electrophoresis (using formaldehyde) followed by ethidium bromide staining revealed fluorescent bands expected for undegraded bacterial rRNA.
Numerous modifications and variations of the present invention are included in the above-identified specification and are expected to be obvious to one of skill in the art. For example, other appropriate sources of nucleic acids may be employed in the performance of the present method, as well as other optional agents. Such modifications and alterations to the compositions and processes of the present invention are believed to be encompassed in the scope of the claims appended hereto.
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Scion xD Problem Report
Scion xD Abnormal Noise or Vibration on Acceleration
(18 reports)
An abnormal noise or vibration on acceleration from rest or on gear shift may develop. Our technicians tell us a redesigned left side engine mount insulator is available to address this problem.
Manual 5 speed will evidence a distinct higher pitch noise or whistle every time I accelerate. Independent of the clutch, and always stops the second I decelerate. Local Toyota/Scion dealership's mechanic did not hear the sound. So, I asked to ride with one to see if it occurred while in the car together. Mac, the service rep. did ride with me, the noise was clearly there - subtle, but clearly there - and he did not hear it. I plan on continued web research and a follow up visit to this or another dealer. It's real, and it's under warranty. I will not let this one go. - Visitor
2 people like this
I HAVE THE SAME PROBLEM, WHEN ACCELERATE THE ENGINE START MAKING ABNORMAL NOISE AND VIBRATION.?? I WONDER WHAT CAUSE THAT? - Visitor
1 person likes this
Very strong vibration while under throttle in 1st and 2nd, more than 1/4 to 1/3 throttle. 5 speed manual. 3rd gear, hardly an issue unless merging at WOT - tmrepp
1 person likes this
Small whistle noise under slight acceleration, silent thereafter. Dealership says waterpump and belt. I will update next week following my apt. - Visitor
Vibration after start up and moving between first and second on automatic transmission. - Visitor
Noise that eventually became a rattle. Then I couldn't accelerate any more. Trying to get it fixed now. - Visitor
The bolt at the top of the motor mount insulator sheered off. I am having a very difficult time finding a replacement bolt. The diagrams online do not show this bolt. They do show the adjacent bolt that connects the stay to the engine, but that bolt is significantly smaller than the bolt at the top of the insulator. The insulator bolt takes a 17mm socket. - Visitor
low pitch noise the increases on acceleration especially when turning to the left - Visitor
abnormal whining noise in all gears and idle - Visitor
abnormal noise and vibration on acceleration, also have rattle noise. Very annoying and as often as ive had it in no fix as of yet. Not happy with this one bit.... - Visitor
rapid clicking noise at mild to moderate acceleration,no fix,no money! alot of heat emanating through firewall,cooking feet and legs. - Visitor
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subset: commoncrawl


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art, Comme des Garcons, Hanae Mori, Japanese designers, Rei Kawakubo, shoes, Tetsuya Uenobe, Tokio Kumagai, Wearable art, Yohji Yamamoto
Kristen Stewart Red Carpet Misstep With Christian Louboutin Shoes That Didn't Fit at the Twilight Eclipse Premier
"It or Miss: If the shoe doesn't fit, don't wear it, even if they are by Christian Louboutin!Kristen Stewart was having her own version of a Cinderella moment on the red carpet last night at "The Twilight Saga: Eclipse" premiere in Los Angeles.Her fabulous Christian Louboutin 'Big Lips Pumps' in beige suede looked like they were about a size too large.>> SHOP for Dress Pumps in your size!As a matter of fact, we usually love Kristen's off-handed style, but last night was a bit of a fit miss for her. In our opinion, her Elie Saab one-shoulder mini was a bit long in its single sleeve and didn't do wonders for her figure.originalMore celebrity shoe porn from last night because we know that you cannot resist a fab pair of stilettos…"Published on June 25, 2010
Amber Rose, Anna Kendricks, celebrity, Christian Louboutin, Christian Louboutin Big Lips Pumps, Dakota Fanning, Elie Saab, it or miss, Jaime King. Nikki Reed, Kristen Stewart, red carpet, red carpet shoes, shoe porn, shoes, The Twilight Saga: Eclipse, Twilight, Twilight Eclipse
Best platform wedge sandals comfy enough for diehard flat-wearers
"How to find a flattering shoe with a bit of 'lift' when you are a flats devotee.Hey Sharon– love your piece on the best platform shoes of the season. I, however, am a chronic flat-wearer; the height of my footwear varies only if I wear thick socks with my Adidas.Which are the best (ie easiest) platform pairs to wear that won't send me to my local podiatrist … know any good ones?!- (New York, NY via e-mail)"Published on May 18, 2010
comfy shoes, Dr. Michael Scanlon, flats, platform, podiatrist, Robert Clergerie, shoe tips, shoes, STYLIST ADVICE, wedge
Cool Clogs
Shopping Guide: Cool Clogs Holy clog, the classic Swedish clog has been fashionably updated! What's a better way to kick back after a night in sexy stilettos than with a pair of super comfortable clogs? Yes, Matilda, the country classic comfort clog is back and revved up as a trendy fashion shoe with plenty of…


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ASG STI 1911-A1 RSS Shell Ejecting Gun
Sign up to get notified when this product is back in stock
Availability: Out of Stock
Product Code:GP-ASG-191-0010
Manufacturer:Action Sport Games (ASG)
- Velocity: 280 fps (0.2 g BB) / Range 100-150 feet
- Barrel Length: 5 inches / 17 cm
- Magazine Capacity: 8 Rounds. 1 Per Shell
- Authentic STI Markings
-Manufactured by Marushin
- 10 Shells Included
The first realistic shooting simulation (RSS) pistol based on the 1911 design. The magazine holds 8 cartridges loaded into the magazine like a real firearm - each cartridge holds one 6mm BB. When fired, the spent cartridge will eject like a real cartridge, and a new cartridge is loaded into the chamber of the pistol. The spent cartridges can be used over and over again by simply loading them with a new 6mm BB. The 1911 pistol is powered with gas which is stored in the magazine. The technology behind the realistic shooting simulation is based on the patented technology of Marushin Kogyo Co. LTD, Japan.
Additional Information
|FPS Range
|Action Sport Games (ASG)
|PreOrder Lead Time in Weeks
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Redbirds storm into PCL playoffs against Omaha after strong regular-season finish
During the dog days of July, it looked like the Redbirds would have to fight and claw to the very end to even have a chance at winning the Pacific Coast League's American Southern Division.
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Gunmetal Black Ep 01
episode 14.26 / 5.00 4,983 Views
Lucina's First Day
A look at the first day of Lucina in Super Smash4.24 / 5.00 15,959 Views
BATMAN v SUPERMAN final showdown
SPOILERS!!4.19 / 5.00 14,594 Views
Another great flash. I liked the added Weird Al song in the back. Funny too, especially the stereotype comments.
I liked how you had Weird Al playing in the background
That was funny as hell!
that shit was
funny as the the devil rapes u violently
The wonderful thing about Tiggers is I'm the only one! That was a nice cover-up.


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- Kyle Bonagura, ESPN Staff Writer
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STANFORD, Calif. -- Steve Sarksian walked out of the Washington locker room with a smile on his face. His demeanor showed nothing to indicate any frustration as a result of the controversial finish to his team's 31-28 loss to Stanford.
This, despite his astonishment in the replay official's decision to overturn a pass that would have given Washington a first down at the Stanford 33 with less than two minutes left. After viewing the replay on video board, Sarkisian didn't think there was enough evidence to reverse what was initially ruled a 16-yard reception for Kevin Smith on fourth down.
"It was ruled a catch, and to determine it not a catch they had to overrule it and determine it wasn't," Sarkisian said. "And from my vantage point, it looked like it was pretty hard to overturn it. Again, I didn't get to sit 50 yards up in the booth and play a video game and make a call."
In real time, Stanford coach David Shaw saw it the same way. His coaches in the booth did not.
"They saw a replay from behind, especially, the view that I didn't have, you could see the ball go through, hit his chest, bounce off the ground and into his hands," Shaw said.
That was the explanation relayed by the lead official, a ruling that allowed Stanford to kneel twice and hold on for the win.
"It's unfortunate the game had to come down to a judgment call like that," Sarkisian said. "That part was unfortunate, because it was two good football teams battling and competing with one another, and I wish the game would have gotten won on the field and not in the booth upstairs with some guy that didn't get to feel the emotion and the hard fought football game that that game was."
A win against No. 5 Stanford (5-0, 3-0 Pac-12) would easily have been the biggest win of Sarkisian's career and perhaps vaulted the No. 15 Huskies (4-1, 1-1) into the top 10. Instead, they'll settle for heartbreak.
"There's no awards for losing," Sarkisian said. "We're a proud football team, and we're proud of the way we played, but we came here to win the game, and unfortunately, we didn't do that."
The Huskies will shift their attention quickly to No. 2 Oregon, which travels to Husky Stadium next week.
Happy Fourth of July from the Pac-12 Blog
17hChantel Jennings
Mailbag: Awkward Jim Harbaugh, Baylor's schedule
1dTed Miller
The replacements: Washington State
1dDavid Lombardi
More on arrest of Utah CB Dominique Hatfield
2dTed Miller
Best wins of coaches' Pac-12 careers: South
2dChantel Jennings
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2dTravis Haney
Oregon among best at building a national brand
2dChantel Jennings
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2dChantel Jennings
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Despite loss, Huskies don't feel beaten


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According to Dr. Max of The Tooth Truth, symptoms of 12-year molar eruption are similar to those of primary teeth eruption and include pain, swelling and inflammation of gum tissue at the site of eruption. Ectopic eruptions are a less common but more serious problem afflicting a small percentage of children developing 12-year molars.Know More
Carla Cohn of Oral Health Group reports different literature placing the frequency of ectopically erupted molars at between 0.75 percent to 6 percent of the population. The frequency is 25 percent for children with cleft lips. Children whose siblings have suffered from the condition are five times more likely to get ectopically erupted molars. According to Cohn, some signs of ectopic eruptions include asymmetric development, secondary molars growing at the wrong angles and prematurely loose primary molars.
There are different ways of dealing with the symptoms. According to Dr. Max, pain and inflammation can be treated with nonsteroidal anti-inflammatory medications such as ibuprofen. The eruption site should be kept clean, and foods that can fragment, such as nuts and popcorn, should be avoided. Infected eruption sites are treated with antibiotics. Patients with ectopically erupted 12-year molars need to see a dentist. Depending on the severity of the condition, ectopic eruptions are treated with separators, surgery or by extraction of affected teeth, says Cohn.Learn more about Dental
The second primary molars are lost around 10 or 11 years of age to make way for the permanent second molars, according to Children's Hospital of Wisconsin. The permanent second molars typically erupt around 12 years of age. The second primary molars are generally the last baby teeth lost.Full Answer >
Children typically lose their first and second primary molars between the ages of 6 and 12. Usually children lose their incisors before losing their cuspids or molars.Full Answer >
The amount of time it will take for pain to last after getting fillings is subjective, but it can take up to a few weeks, as stated by WebMD. Pain relievers may not be necessary to help with the pain.Full Answer >
Treatment for toothache depends on the cause; a toothache caused by a cavity may require filling or extraction of the affected tooth, and a toothache caused by infection is generally treated with antibiotics. According to WebMD, a root canal may be needed if the toothache is caused by an infection on the nerve of the tooth.Full Answer >


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BarCampNola 8
- Science & Technology
- 6823 St Charles Ave, New Orleans, LA, 70118
YP National Day of Service: STEM Project
Urban League GNO Young Professionals
Friday, April 25, 2014 from 8:00 AM to 12:00 PM (CDT)
Jobs in S.T.E.M. related fields (Science, Technology, Engineering and Math) offer a wealth of opportunities not only currently but for generations to come. Unfortunately, African-Americans are under-represented in these fields. Often because we are not aware of the opportunities that exist.
This year, YP is again partnering with the National Society of Black Engineers (NSBE) to host a STEM project at one of our area schools to encourage our kids to explore opportunities in these fields. Join us at the Discovery Health Sciences School in Kenner for a day of education and fun with our kids.
This year's competition will focus on wind energy. Teams will compete to create the most effective windmill. Prizes will be awarded to the winning team.
To make this program a success we need VOLUNTEERS. Join us on Friday morning, April 25th from 8 am to 12 noon to support our kids and introduce them to new career options. Any questions, you can send an email to email@example.com. Also, don't forget to wear your YP t-shirt or a red or black t-shirt representing our YP colors.
|The Mission
|The Urban League of Greater New Orleans Young Professionals ("ULGNOYP") is an Urban League of Greater New Orleans auxiliary of young professionals committed to promoting and supporting the ideas and goals of the Urban League movement throughout the New Orleans area through community outreach, fundraising and membership development.
Join millions of people on Eventbrite.
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La Flora Resort Patong
- Free WiFi and free parking
Main amenities
- 67 guestrooms
- On the beach
- 2 restaurants
- Full-service spa
- 2 outdoor pools
- Breakfast available
- Fitness center
- Beach umbrellas
- Business center
- Airport shuttle
- 24-hour front desk
- Air conditioning
What's around
- On the beach
- Patong Beach (0 mi / 0.1 km)
- Bangla Night Street (0.2 mi / 0.3 km)
- Sphinx Theatre (0.5 mi / 0.8 km)
- Jungceylon Shopping Center (0.5 mi / 0.8 km)
- Bangla Boxing Stadium (0.6 mi / 1 km)
We booked for a honeymoon trip and it was so noisy ...we hear everything around and men…
I was stayed with a friend, unfortunately hotel only has single king size beds, which was…
Enter dates to check prices
Hotel Info: 01 65 69 441from €172
- Pool Villa With Loft
- Pool Villa
- Beachfront Grand Pool Villa
- Deluxe Room, Pool View
- Deluxe Room, Pool Access
- Sea View Suite
- Beachfront Pool Villa
Recommended hotels
At a glance
Key facts
Hotel size
- This hotel has 67 rooms
- This hotel is arranged over 2 floors
- Check-in time starts at 2 PM
- Check-out time is noon
Required at check in
- Credit card or cash deposit required
- Government-issued photo ID required
- Minimum check-in age is 18
Travelling with others
- No pets or service animals allowed
- Free WiFi in public areas
- Free WiFi in rooms
- Airport shuttle (available on request) *
- Free self parking (limited)
Nearby Four Star Phuket - Phangnga hotels
In the hotel
- Buffet breakfast daily (surcharge)
- 2 restaurants
- 24-hour room service
- Beach cabanas (surcharge)
- Beach sun loungers
- Number of outdoor pools - 2
- Pool sun loungers
- Fitness facilities
- Full-service spa
- Spa treatment room(s)
- Scooter/moped rentals nearby
- Beach umbrellas
- Pool umbrellas
- Steam room
- Beach towels
- Sauna
- Business center
- Number of meeting rooms - 2
- Meeting rooms
- Computer station
- 24-hour front desk
- Concierge services
- Limo or Town Car service available
- Dry cleaning/laundry service
- Free newspapers in lobby
- Number of buildings/towers - 3
- Safe-deposit box at front desk
- Designated smoking areas
- Library
- Television in lobby
In the room
- Air conditioning
- Minibar
- Coffee/tea maker
- Bathrobes
- Iron/ironing board (on request)
- Blackout drapes/curtains
- Turndown service
- Premium bedding
- Individually furnished
- Balcony
- Private bathroom
- Deep soaking bathtub
- Free toiletries
- Hair dryer
- Satellite TV channels
- DVD player
- Free newspaper
- Free WiFi
- Phone
- Refrigerator
- Free bottled water
- Daily housekeeping
- In-room safe
Top Phuket - Phangnga hotels
Recently reviewed hotels in Phuket - Phangnga
- Kalima Resort & Spa, PhuketFrom
"Enjoyable"Jun 21, 2015
- Outrigger Laguna Phuket Beach ResortFrom
"I stayed in Outrigger for 5 nights. From 25 till 30th of May. The resort has a beautiful nature with lagoon view. I like a free shuttle boat which can take you free around the…"Jun 21, 2015
- Holiday Inn Express Phuket Patong Beach CentralFrom
"we spent 6 nights in the hotel, and it's amazing hotel with a good location from the beach."Jun 20, 2015
- Noble HouseFrom
"i very enjoy my holidays in this hotel, the good people and the good staff. I hope return in here. Thank you for all."Jun 17, 2015
Nearby hotels
Special features
Flora Spa has massage/treatment rooms. Services include massages, facials, and body treatments. The spa is equipped with a spa tub. A variety of treatment therapies are provided, including aromatherapy.
DiningThe Current of the Sea - This restaurant specializes in Thai cuisine and serves breakfast, lunch, and dinner.
The Surface - This restaurant serves breakfast, lunch, and dinner. Guests can order drinks at the bar and enjoy alfresco dining (weather permitting).
La Flora Resort Patong's small print
This property does not have elevators.
The name on the credit card used at check-in to pay for incidentals must be the primary name on the guestroom reservation.
Mandatory fees
You'll be asked to pay the following charges at check in or check out:
- New Year's Eve (31 December) Gala Dinner per adult: THB 6500
- New Year's Eve (31 December) Gala Dinner per child: THB 3250 (from 5 to 12 years old)
Optional extras
Rollaway beds are available for THB 1500 per night
Buffet breakfast is offered for an extra charge of THB 450.00 per person (approximately)
Airport shuttle service is offered for an extra charge of THB 1000 per vehicle (one way)
We have included all charges provided to us by this hotel. However, charges can vary, for example, based on length of stay or the room you book.


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Most Popular "Coney Island Brooklyn New York City" Titles
- Movies or TV
- Genres
- Keywords
- IMDb Rating
- Instant Watch Options
- In Theaters
- On TV
- Release Year
1. Requiem for a Dream (2000)
R | 102 min | Drama
The drug-induced utopias of four Coney Island people are shattered when their addictions become stronger.
Writers: Hubert Selby Jr., Darren Aronofsky | Director: Darren Aronofsky | Stars: Ellen Burstyn, Jared Leto, Jennifer Connelly, Marlon Wayans
Votes: 504,685
2. Mr. & Mrs. Smith (2005)
PG-13 | 120 min | Action, Comedy, Crime
A bored married couple is surprised to learn that they are both assassins hired by competing agencies to kill each other.
Writer: Simon Kinberg | Director: Doug Liman | Stars: Brad Pitt, Angelina Jolie, Adam Brody, Vince Vaughn
Votes: 302,483
3. Men in Black 3 (2012)
PG-13 | 106 min | Action, Comedy, Sci-Fi
Agent J travels in time to M.I.B.'s early days in 1969 to stop an alien from assassinating his friend Agent K and changing history.
Writers: Etan Cohen, Lowell Cunningham | Director: Barry Sonnenfeld | Stars: Will Smith, Tommy Lee Jones, Josh Brolin, Jemaine Clement
Votes: 236,603
4. A.I. Artificial Intelligence (2001)
PG-13 | 146 min | Adventure, Drama, Sci-Fi
A highly advanced robotic boy longs to become "real" so that he can regain the love of his human mother.
Writers: Brian Aldiss, Ian Watson, Steven Spielberg | Director: Steven Spielberg | Stars: Haley Joel Osment, Jude Law, Frances O'Connor, Sam Robards
Votes: 218,037
5. The Warriors (1979)
R | 92 min | Action, Thriller
In 1979 a charismatic leader summons the street gangs of New York City in a bid to take it over. When he is killed, The Warriors are falsely blamed and now must fight their way home while every other gang is hunting them down to kill them.
Writers: Sol Yurick, David Shaber, Walter Hill | Director: Walter Hill | Stars: Michael Beck, James Remar, Dorsey Wright, Brian Tyler
Votes: 63,977
6. American Gangster (2007)
R | 157 min | Biography, Crime, Drama
In 1970s America, a detective works to bring down the drug empire of Frank Lucas, a heroin kingpin from Manhattan, who is smuggling the drug into the country from the Far East.
Writers: Steven Zaillian, Mark Jacobson | Director: Ridley Scott | Stars: Denzel Washington, Russell Crowe, Chiwetel Ejiofor, Josh Brolin
Votes: 302,448
7. Cloverfield (2008)
PG-13 | 85 min | Adventure, Sci-Fi, Thriller
A group of friends venture deep into the streets of New York on a rescue mission during a rampaging monster attack.
Writer: Drew Goddard | Director: Matt Reeves | Stars: Mike Vogel, Jessica Lucas, Lizzy Caplan, T.J. Miller
Votes: 259,402
8. Empire of the Sun (1987)
PG | 153 min | Drama, History, War
A young English boy struggles to survive under Japanese occupation during World War II.
Writers: J.G. Ballard, Tom Stoppard | Director: Steven Spielberg | Stars: Christian Bale, John Malkovich, Miranda Richardson, Nigel Havers
Votes: 81,517
9. Annie Hall (1977)
PG | 93 min | Comedy, Drama, Romance
Neurotic New York comedian Alvy Singer falls in love with the ditsy Annie Hall.
Writers: Woody Allen, Marshall Brickman | Director: Woody Allen | Stars: Woody Allen, Diane Keaton, Tony Roberts, Carol Kane
Votes: 167,785
10. Butch Cassidy and the Sundance Kid (1969)
M | 110 min | Biography, Crime, Western
Two Western bank/train robbers flee to Bolivia when the law gets too close.
Writer: William Goldman | Director: George Roy Hill | Stars: Paul Newman, Robert Redford, Katharine Ross, Strother Martin
Votes: 132,774
11. He Got Game (1998)
R | 136 min | Drama, Sport
A basketball player's father must try to convince him to go to a college so he can get a shorter sentence.
Writer: Spike Lee | Director: Spike Lee | Stars: Denzel Washington, Milla Jovovich, Ray Allen, Rosario Dawson
Votes: 33,650
12. Very Good Girls (2013)
R | 91 min | Drama
Two New York City girls make a pact to lose their virginity during their first summer out of high school. When they both fall for the same street artist, the friends find their connection tested for the first time.
Writer: Naomi Foner | Director: Naomi Foner | Stars: Dakota Fanning, Elizabeth Olsen, Sterling Jones, Lenny Platt
Votes: 7,492
13. Two Weeks Notice (2002)
PG-13 | 101 min | Comedy, Romance
A lawyer decides that she's used too much like a nanny by her boss, so she walks out on him.
Writer: Marc Lawrence | Director: Marc Lawrence | Stars: Sandra Bullock, Hugh Grant, Alicia Witt, Dana Ivey
Votes: 77,848
14. Nine 1/2 Weeks (1986)
R | 117 min | Drama, Romance
A woman gets involved in an impersonal affair with a man. She barely knows about his life, only about the sex games they play, so the relationship begins to get complicated.
Writers: Sarah Kernochan, Zalman King, Patricia Louisianna Knop, Elizabeth McNeill | Director: Adrian Lyne | Stars: Mickey Rourke, Kim Basinger, Margaret Whitton, David Margulies
Votes: 27,024
15. Jeanne Eagels (1957)
108 min | Biography, Drama
The life of 1920s actress Jeanne Eagels.
Writers: Daniel Fuchs, Sonya Levien, John Fante | Director: George Sidney | Stars: Kim Novak, Jeff Chandler, Agnes Moorehead, Charles Drake
Votes: 490
16. Pi (1998)
R | 84 min | Drama, Thriller
A paranoid mathematician searches for a key number that will unlock the universal patterns found in nature.
Writers: Darren Aronofsky, Sean Gullette, Eric Watson | Director: Darren Aronofsky | Stars: Sean Gullette, Mark Margolis, Ben Shenkman, Pamela Hart
Votes: 132,720
17. Angel Heart (1987)
X | 113 min | Horror, Mystery, Thriller
Harry Angel is a private investigator. He is hired by a man who calls himself Louis Cyphre to track down a singer called Johnny Favorite. But the investigation takes an unexpected and somber turn.
Writers: William Hjortsberg, Alan Parker | Director: Alan Parker | Stars: Mickey Rourke, Robert De Niro, Lisa Bonet, Charlotte Rampling
Votes: 57,907
18. New York, I Love You (2008)
R | 103 min | Comedy, Drama, Romance
Several love stories set in one of the most loved cities of the world, New York.
Writers: Hu Hong, Yao Meng, Israel Horovitz, Suketu Mehta, Shunji Iwai, Olivier Lécot, Jeff Nathanson, Xan Cassavetes, Stephen Winter, Anthony Minghella, Natalie Portman, Fatih Akin, Joshua Marston, Hall Powell, James C. Strouse, Emmanuel Benbihy, Tristan Carné | Directors: Fatih Akin, Yvan Attal, Randall Balsmeyer, Allen Hughes, Shunji Iwai, Wen Jiang, Shekhar Kapur, Joshua Marston, Mira Nair, Natalie Portman, Brett Ratner | Stars: Shia LaBeouf, Natalie Portman, Bradley Cooper, Hayden Christensen
Votes: 37,158
19. The Taking of Pelham 1 2 3 (2009)
R | 106 min | Action, Crime, Thriller
Armed men hijack a New York City subway train, holding the passengers hostage in return for a ransom, and turning an ordinary day's work for dispatcher Walter Garber into a face-off with the mastermind behind the crime.
Writers: Brian Helgeland, John Godey | Director: Tony Scott | Stars: Denzel Washington, John Travolta, Luis Guzmán, Victor Gojcaj
Votes: 135,952
20. Thick as Thieves (I) (2009)
R | 104 min | Action, Crime, Thriller
A veteran thief recruits a younger crook to help him pull off one final job in order to repay his debt to the Russian mob.
Writer: Ted Humphrey | Director: Mimi Leder | Stars: Rade Serbedzija, Morgan Freeman, Robert Forster, Corey Johnson
Votes: 26,869
21. Gentlemen Prefer Blondes (1953)
Not Rated | 91 min | Comedy, Musical, Romance
Singers Lorelei Lee and Dorothy Shaw travel to Paris, pursued by a private detective hired by the disapproving father of Lorelei's fiancé to keep an eye on her, as well as a rich, enamored old man and many other doting admirers.
Writers: Charles Lederer, Joseph Fields, Anita Loos | Director: Howard Hawks | Stars: Jane Russell, Marilyn Monroe, Charles Coburn, Elliott Reid
Votes: 20,287
22. Uptown Girls (2003)
PG-13 | 92 min | Comedy, Drama, Romance
A grown-up woman, who kept her childish instincts and behavior, starts working as a nanny of a 8-year-old girl, who actually acts like an adult. But in the end everything turns to its right places.
Writers: Julia Dahl, Mo Ogrodnik, Lisa Davidowitz, Allison Jacobs | Director: Boaz Yakin | Stars: Brittany Murphy, Dakota Fanning, Heather Locklear, Jesse Spencer
Votes: 26,803
23. Death to Smoochy (2002)
R | 109 min | Comedy, Crime, Drama
Kids show host Rainbow Randolph is fired in disgrace while his replacement, Sheldon Mopes, aka Smoochy the Rhino, finds himself a rising star. Unfortunately for Sheldon, the kid's TV business isn't all child's play.
Writer: Adam Resnick | Director: Danny DeVito | Stars: Robin Williams, Edward Norton, Catherine Keener, Danny DeVito
Votes: 33,404
24. Imitation of Life (1959)
Not Rated | 125 min | Drama, Romance
A struggling young actress with a six-year-old daughter sets up housekeeping with a homeless black widow and her light-skinned eight-year-old daughter who rejects her mother by trying to pass for white.
Writers: Eleanore Griffin, Allan Scott, Fannie Hurst | Director: Douglas Sirk | Stars: Lana Turner, John Gavin, Sandra Dee, Susan Kohner
Votes: 10,023
25. The Pick-up Artist (1987)
PG-13 | 81 min | Comedy, Crime, Drama
A womanizer meets his match when he falls for a woman in debt to the mafia.
Writer: James Toback | Director: James Toback | Stars: Molly Ringwald, Robert Downey Jr., Dennis Hopper, Danny Aiello
Votes: 4,460
26. On the Town (1949)
Passed | 98 min | Comedy, Musical, Romance
Three sailors on a day of shore leave in New York City look for fun and romance before their twenty-four hours are up.
Writers: Adolph Green, Betty Comden, Jerome Robbins | Directors: Stanley Donen, Gene Kelly | Stars: Gene Kelly, Frank Sinatra, Betty Garrett, Ann Miller
Votes: 11,079
27. Radio Days (1987)
PG | 88 min | Comedy
A nostalgic look at radio's golden age focusing on one ordinary family and the various performers in the medium.
Writer: Woody Allen | Director: Woody Allen | Stars: Mia Farrow, Dianne Wiest, Mike Starr, Paul Herman
Votes: 21,888
28. Heaven Help Us (1985)
R | 104 min | Comedy, Drama, Romance
A new transfer student to St. Basil's Boys' Prep School tries to fit in while romantically pursuing a troubled young girl.
Writer: Charles Purpura | Director: Michael Dinner | Stars: Donald Sutherland, Andrew McCarthy, John Heard, Mary Stuart Masterson
Votes: 3,023
29. The Devil and Miss Jones (1941)
Approved | 92 min | Comedy, Romance
A tycoon goes undercover to ferret out agitators at a department store, but gets involved in their lives instead.
Writer: Norman Krasna | Director: Sam Wood | Stars: Jean Arthur, Robert Cummings, Charles Coburn, Edmund Gwenn
Votes: 2,429
30. The Warriors (2005 Video Game)
M | Action, Crime
The Warriors are a gang from Coney Island. They go to the biggest gang leader's meeting under a truce. But all doesn't go well as the gang leader is shot and killed. Framed for this murder,... See full summary »
Writers: Dan Van Zant, John Zurhellen, Mike Zoccano | Directors: Navid Khonsari, John Zurhellen | Stars: James Remar, Ephraim Benton, Dorsey Wright, David Harris
Votes: 1,002
31. Houdini (1953)
Approved | 106 min | Biography, Drama
The spectacular but tragically short career of magician and illusionist Harry Houdini whose tricks defied explanation and safety.
Writers: Philip Yordan, Harold Kellock | Director: George Marshall | Stars: Tony Curtis, Janet Leigh, Torin Thatcher, Angela Clarke
Votes: 2,100
32. Mutual Friends (2013)
Surprise parties rarely go well. This one is no different, as a group of 30-somethings head through life, love and a day in New York City.
Writers: Francisco Angones, Jessica Sue Burstein, Craig DiFolco, Amy Higgins, Ross Partridge, Olivia Silver, Matthew Watts | Director: Matthew Watts | Stars: Caitlin FitzGerald, Cheyenne Jackson, Peter Scanavino, Michael Stahl-David
Votes: 61
33. The Sting II (1983)
PG | 102 min | Comedy, Crime
Lonnegan is planning to get back at Gondorff and Hooker for bilking him out of half a million. At the same time Gondorff and Hooker are planning their next caper. And their mark is a man ... See full summary »
Writer: David S. Ward | Director: Jeremy Kagan | Stars: Jackie Gleason, Mac Davis, Teri Garr, Karl Malden
Votes: 1,459
34. Enemies: A Love Story (1989)
R | 119 min | Comedy, Drama, Romance
A ghostwriter finds himself romantically involved with his current wife, a married woman and his long-vanished wife.
Writers: Isaac Bashevis Singer, Roger L. Simon, Paul Mazursky | Director: Paul Mazursky | Stars: Ron Silver, Anjelica Huston, Lena Olin, Malgorzata Zajaczkowska
Votes: 1,569
35. Possessed (1931)
Passed | 76 min | Drama, Romance
Ambitious factory Girl Marion Martin meets a handsome well-to-do, but he's interested in her as a mistress, not a wife.
Writers: Edgar Selwyn, Lenore J. Coffee | Director: Clarence Brown | Stars: Joan Crawford, Clark Gable, Wallace Ford, Richard 'Skeets' Gallagher
Votes: 1,031
36. Hell Up in Harlem (1973)
R | 94 min | Crime, Drama
A gangster in Harlem must rescue his ex-wife, who has been kidnapped by the Mafia.
Writer: Larry Cohen | Director: Larry Cohen | Stars: Fred Williamson, Julius Harris, Gloria Hendry, Margaret Avery
Votes: 829
37. Madigan (1968)
101 min | Crime, Drama
Policemen Bonaro and Madigan lose their guns to fugitive Barney Benesch. As compensation, the two NYC detectives are given a weekend to bring Benesch to justice. While Bonaro and Madigan ... See full summary »
Writers: Howard Rodman, Abraham Polonsky, Richard Dougherty | Director: Don Siegel | Stars: Richard Widmark, Henry Fonda, Inger Stevens, Harry Guardino
Votes: 2,115
38. Virgil Bliss (2001)
93 min | Crime, Drama, Romance
Virgil Bliss is a man obsessed with living a normal life, finding a good job, marrying a decent woman, and building a family of his own. But as a recently paroled career thief with a ... See full summary »
Writer: Joe Maggio | Director: Joe Maggio | Stars: Clint Jordan, Kirsten Russell, Anthony Gorman, Greg Amici
Votes: 219
39. The Crowd (1928)
Not Rated | 98 min | Drama, Romance
The life of a man and woman together in a large, impersonal metropolis through their hopes, struggles and downfalls.
Writers: King Vidor, John V.A. Weaver, Joseph Farnham | Director: King Vidor | Stars: Eleanor Boardman, James Murray, Bert Roach, Estelle Clark
Votes: 5,201
40. The Great New Wonderful (2005)
R | 87 min | Comedy, Drama, Romance
The Great New Wonderful weaves five stories against the backdrop of an anxious and uncertain post-9-11 New York City.
Writer: Sam Catlin | Director: Danny Leiner | Stars: Olympia Dukakis, Jim Gaffigan, Judy Greer, Maggie Gyllenhaal
Votes: 1,288
41. Bad Girl (1931)
90 min | Drama
A man and woman, skeptical about romance, nonetheless fall in love and are wed, but their lack of confidence in the opposite sex haunts their marriage.
Writers: Viña Delmar, Brian Marlow, Edwin J. Burke | Director: Frank Borzage | Stars: James Dunn, Sally Eilers, Minna Gombell, Frank Austin
Votes: 525
42. My Friend Irma (1949)
Passed | 103 min | Comedy, Romance
Prototype dumb blonde Irma and her slacker, wheeler-dealer boyfriend Al interfere in the love life of Irma's level-headed room mate Jane.
Writers: Cy Howard, Parke Levy | Director: George Marshall | Stars: John Lund, Diana Lynn, Don DeFore, Marie Wilson
Votes: 603
43. Speedy (1928)
85 min | Action, Comedy, Family
"Speedy" loses his job as a soda-jerk, then spends the day with his girl at Coney Island. He then becomes a cab driver and delivers Babe Ruth to Yankee Stadium, where he stays to see the ... See full summary »
Writers: John Grey, Lex Neal, Howard Emmett Rogers | Director: Ted Wilde | Stars: Harold Lloyd, Ann Christy, Bert Woodruff, Brooks Benedict
Votes: 1,717
44. Anna Christie (I) (1930)
Passed | 89 min | Drama, Romance
A young woman reunites with her estranged father and falls in love with a sailor, but struggles to tell them about her dark past.
Writers: Frances Marion, Eugene O'Neill | Director: Clarence Brown | Stars: Greta Garbo, Charles Bickford, George F. Marion, Marie Dressler
Votes: 1,865
45. Little Fugitive (1953)
Not Rated | 80 min | Drama, Family
Young boy fears he shot his older brother who is only faking. Young boy runs away to Coney Island, a crowded beach resort, gets money by returning soda pop bottles for their deposits.
Writers: Ray Ashley, Morris Engel, Ruth Orkin | Directors: Ray Ashley, Morris Engel, Ruth Orkin | Stars: Richard Brewster, Winifred Cushing, Jay Williams, Will Lee
Votes: 1,406
46. City for Conquest (1940)
Approved | 104 min | Crime, Drama, Music
Cagney is Danny Kenny, a truck driver who enters "the fight game" and Sheridan plays his girlfriend, Peggy. Danny realizes success in the ring and uses his income to pay for his brother ... See full summary »
Writers: John Wexley, Aben Kandel | Directors: Anatole Litvak, Jean Negulesco | Stars: James Cagney, Ann Sheridan, Frank Craven, Donald Crisp
Votes: 1,478
47. Went to Coney Island on a Mission from God... Be Back by Five (1998)
R | 94 min | Drama
Two young men (Jon Cryer, Rick Stear) who have been friends since early childhood decide to go on a trip to find a third friend (Rafael Baez) who has long since disappeared. Stories they ... See full summary »
Writers: Jon Cryer, Richard Schenkman | Director: Richard Schenkman | Stars: Jon Cryer, Rick Stear, Rafael Báez, Ione Skye
Votes: 241
48. Mannequin (1937)
Passed | 95 min | Drama
Rags-to-riches Hennessey meets newlyweds Jessie and Eddie from his old neighborhood. Eddie plots to have Jessie divorce him, marry Hennessey, divorce Hennessey, then bring Hennessey's money... See full summary »
Writers: Lawrence Hazard, Katharine Brush | Director: Frank Borzage | Stars: Joan Crawford, Spencer Tracy, Alan Curtis, Ralph Morgan
Votes: 661
49. Lonesome (1928)
Not Rated | 69 min | Comedy, Drama, Romance
Two lonely people in the big city meet and enjoy the thrills of an amusement park, only to lose each other in the crowd after spending a great day together. Will they ever see each other again?
Writers: Mann Page, Edward T. Lowe Jr., Tom Reed | Director: Pál Fejös | Stars: Barbara Kent, Glenn Tryon, Fay Holderness, Gusztáv Pártos
Votes: 1,222
50. Off Jackson Avenue (2008)
80 min | Crime, Drama, Thriller
Off Jackson Avenue is an interwoven crime story set in New York City involving a Mexican woman who has been tricked into sex-slavery by an Albanian pimp and must find a way to break out; A ... See full summary »
Writer: John-Luke Montias | Director: John-Luke Montias | Stars: Jessica Pimentel, Stivi Paskoski, Jun Suenaga, John-Luke Montias
Votes: 28


text:
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Yesterday I encountered my latest pregnancy challenge: the dreaded morning sickness. I woke up nauseated and it got worse as the day went on. The nausea itself wasn't that bad, but I had no appetite, which caused a massive blood-sugar crash that lasted the whole day. It was like having the flu: I was shaking, dizzy, freezing cold, weak, and my muscles ached. But no matter how bad it was, I couldn't get myself to eat more than a few bites of food. Food was revolting. I didn't actually vomit, but it was pretty close.
It's so strange. I had a couple of moments of mild nausea in the past two weeks, but nothing serious. In all of my six previous pregnancies, I've never suffered more than those mild, rare moments of sickness. Yesterday, though, it hit me like a ton of bricks. What changed?
I did try some ginger tea which helps a lot with the nausea, but it doesn't help me want to eat, which is the real problem (I've actually lost a pound in the past month). Also, this tea has a couple of other herbs in it which are on the "forbidden during pregnancy" list. I'm drinking a cup right now, but I can't drink it all day long. There are other products with ginger, but my nurse-friend says you really need higher amounts of ginger than you're going to get in most crackers and things. So I'll probably check out Trader Joe's where they seem to have super-strong ginger products, or even make my own tea from fresh ginger. But I'm not sure how I'm going to find the energy to get to the market at all, let alone cook anything. And that's another problem. I think there might be a few palatable foods out there, but I can't imagine going to the market feeling like I did yesterday. I'm sure I'd pass out.
Today, I have managed to eat a bit more. I also slept until 11am and then took a quick nap after lunch, which I think helped a lot. Maybe the tea also helped, or maybe I'm just having a better day. But I'm not cold, which was the worst part about yesterday. I'd have these episodes where the coldness would start out from my core and just spread throughout my body, ending up in my toes, fingers, and nose. And when it got there it wouldn't leave. I kept running my hands under hot water (I was too weak to take a shower) and when I was home, I got under a blanket with a heating pad on my feet. When I went to bed last night, it took a full hour with the heating pad for me to stop feeling cold.
Today is supposed to be my last day of estrogen and progesterone (8 weeks pregnant today – woohoo!), and stopping the hormones might help. However, since I've been so uptight about miscarriage, my RE suggested that I keep going with them, not because it would lower any risk, but because if I did lose the pregnancy, at least I wouldn't blame it on stopping the hormones, and therefore, on myself. Then my OB said I should keep taking them until eleven weeks. I think his information is just a bit out of date. But still, I was going to keep going with the hormones until I ran out, which will probably be in a week or two. Now, I don't know. Maybe I should stop them sooner. I guess I'll give it a few more days and see how it goes.


text:
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Bondage Professor, Curious Student— They explore their private sexual fantasies together.
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Submission to an Old Friend— An older man finds unexpected depth over coffee.
Jane is Trained to Submit Ch. 10— A visitor takes her.
Seeking New Intensity— An older man with a taste for submission finds new depths.
Learning to Score— Sam submits to Joey's teachings.
Girlfriend Finally Cuckolds Pt. 05— Steve takes Val in his own apartment.
Saturday Slave— A lost flash drive puts an older woman under control.
Caught in Her Panties— Wife sets webcam to record her husband and her suspicions.
The Insurance Physical— Making a deadline leads to an humiliating experience.
Casey's Reunion— Chub puts finishing touches on humiliation of narrator.
Jane is Trained to Submit Ch. 09— Lost in a card game.
Salon Girl Takes Over— A woman finds her identity as a collared slave is dangerous.
Domestic Discipline Ch. 03— Chapter 3.
Mr. Staring Problem— Why didn't I stay for the fireworks? A lone bus trip mistake.
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Dress Off Origins: Jane vs Kate— Two women compete in a game of very public humiliation
Bound Friends Pt. 13— Sophia and Lea get new companionship.
Humiliated by a Co-ed in Public— College senior is humiliated by his younger crush.
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Guess: The Ultimate Sex Game Ch. 05— Danielle and Peter spend some quality time together.
Talya and My Trunks— I am exposed and humiliated.
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Rodney's Nude Humiliation Ch. 07— Forced to model in front of Mom's bridge club.
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A Trip to Atlanta— A woman finds that you have to be careful what you wish for.
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Girlfriend Finally Cuckolds Pt. 03— Girlfriend goes further still.
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Dove's Tale Ch. 04— Can Dove submit?
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More from Dangerous Assignment on Relic Radio Thrillers this week. We'll hear the episode from March 25, 1953, Find Antonina Aquirre.
This week on Legacy Of Laughs, we'll hear from The Bob Hope Show. This episode features guest, Claire Trevor, and aired September 13, 1955.
Case Closed begins with The Crime Club this time. We'll hear Death Swims At Midnight, their story from August 28, 1947. This Is Your FBI follows with, The Jolly Widow. That episode was first heard August 22, 1952.
Theater Five begins the show this week with their story, Annie Is Watching. That episode first aired March 3, 1965. Then, Tales Of The Texas Rangers rides in with their August 10, 1952, story, Last Stop.
Relic Radio Science Fiction features another story from X Minus One this time. From November 28, 1957, here's Shock Troop.
This week on Strange Tales, The Clock presents its tale titled, Behind The Mask. This episode was heard May 1, 1956.
A classic from The Witch's Tale on The Horror this time. We'll hear their story from March 1933, titled, Graveyard Mansion.
This time on Relic Radio Thrillers, The Chase brings us a tale titled, The Apprentice. This episode was first aired July 24, 1952.
We hear from Fibber McGee And Molly on Legacy Of Laughs this week. Here's The Missing Car Fender, which first aired April 1, 1941.
This week on Case Closed, Broadway Is My Beat starts off the show with, The Jimmy Dorn Murder Case from July 7, 1949. Then, Boston Blackie brings us his story from March 12, 1946, Lee Chow Murdered.


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text:
Here's part 2 of last week's blog.
Only one or two
students really blossom
|How many of your students
make it into a flower?
Photo by l'Ours
Why is that? How is it possible that you teach the 3rd conditional, or some tricky 'turn' phrasal verbs, but only one or two students will actually remember, and be able to produce it, in the next class? I guess I've known this for a while, but it's only really this year that I've taken on board the fact that no matter how hard we try to improve our students English, only a couple will really absorb the language we throw at them.
What I noticed this year is that it's always the ones who are avid note takers. You know, the ones who are always asking what everything means, who copy everything you write on the board, who are the ones who use those turn phrasal verbs in their next writing assignment.
What I noticed this year is that it's always the ones who are avid note takers. You know, the ones who are always asking what everything means, who copy everything you write on the board, who are the ones who use those turn phrasal verbs in their next writing assignment.
Wouldn't it be great if everyone was avid note takers, not just for the sake of copying, but because they actually want to learn the word, or maybe be an English teacher one day, go and live in an English speaking country and be able to sing at karaoke without everyone taking the piss out of their accent.
6) Doing a DELTA was a waste of time
6) Doing a DELTA was a waste of time
There, I've said it. After all those blogs I wrote about DELTA, saying how marvellous and life changing it was blah blah blah, now I've gone and done a 360 and decided it was all a load of tosh. Why oh why did I give up a year of my life, my writing, my hair, and torment myself with the pressure of doing a DELTA?
Okay, I'm exaggerating slightly. Of course it wasn't a waste of time and I'm a much better teacher, at least in the eyes of Cambridge, but there was a load of crap on that course, wasn't there?
I mean, really, what was the point in memorising so much terminology for that exam? I spent a good thirty minutes a day memorising words like catenation, onset, and morphology. Who in their right mind would ever use these words while teaching? It's insane.
It's funny but one of the main areas I had to improve on my DELTA was drilling. I still drill in class, don't get me wrong, but not to the extent it was necessary to get a decent mark in Module 2. That brings me on to all the time we needed to prepare one hour of teaching. The reading, preparing, practising that we had to do was so unrealistic to the real world that it all seems a bit silly. I mean who has a spare 10 hours a week to plan for one class? And don't even get me started on Module 3. Planning a course for a bunch of imaginary students I'll never teach…please.
Saying that, if you want a career in TEFL and do want to improve the way your teach, then the DELTA is worth it, just be prepared for jumping through the Cambridge hoops to get there.
7) Teaching phonetics is useless unless you go over it often
7) Teaching phonetics is useless unless you go over it often
I made the same mistake again this year. I've done it two years running now. At the start of term I get all excited about phonetics and firmly believe that it's a great tool for improving student's pronunciation. Or at least showing them visually where they are going wrong. I spent about two weeks with several of my classes going over the chart in detail, doing activities, and drilling. For a few weeks I used phonetics actively in class, did a few songs, and there seemed to be progress. However, as always, exams and syllabus got in the way and pronunciation got pushed to one side in January. Stupidly I never got back into it again this year. The other week I did a few games to try to remind students of the sounds, but it was useless, half of them had forgotten.
So there's your lesson. Don't even bother with phonetics unless you are going to do it regularly. I hope I remember next term.
8) You can live comfortably and support a family as a TEFL teacher in Spain, or at least in Sevilla.
|Hard work never killed anyone...not in TEFL anyway.
Photo by kylesteed
8) You can live comfortably and support a family as a TEFL teacher in Spain, or at least in Sevilla.
After all those years of worrying whether I'd be able to survive while having kids as a TEFL teacher I can now safely say that I can. It's hard work, and I have to do extra business classes early in the morning, which means long days, falling asleep on the metro in the afternoon, and getting home knackered a couple of nights a week, but it's all worth it. Luckily I am a B1 and B2 oral examiner too, which definitely helps get that extra cash over the year to pay for the extras.
We still live quite simply at the moment, we rarely go out in the evenings or travel much, but that's mainly because we two little monsters to look after, but we get by and enjoy life. Hopefully when my wife goes back to work we'll be able to afford a few luxuries too, maybe get a car and have a couple of decent holidays a year.
So if you're wondering about whether you can afford to teach English and have children, then the answer is yes, you can, but you have to work your arse off.
So what have you learnt this year? Enlighten me.


text:
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Thread: USB and External HDD ProblemsTweet
18th October 2011, 22:22 #1
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USB and External HDD Problems
Been playing with my new 800 SE for a few weeks and have tried a few images and for the moment have settled on the Newnigma2 by ******* SSL84.
Eventually thought about a hard drive and tried a 16mb usb drive and an external samsung hdd with its own power supply.
Both gets mounted but will not initialize and then loses format and have to format again. Have tried fat32 and ext3 both from pc via Easeus and within box but always fail to initialize with the following messages.
Partition sda1 mounted on: /autofs/4E91-D793 This can change numbers at end each time I try.
Unable to initialise harddisk.
Error: Mkfs failed
Unable to complete filesystem check.
Error: Fsck failed
Have been trying for a few days now, would like to get them both working before I move onto buying an internal drive for it.
When I try the record button says no hdd found or not initialized. Want to use the usb for epg and external hard drive for recording and stuff.
Any help in right direction and what Im doing wrong would be great. Thanks
18th October 2011, 22:47 #2
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Re: USB and External HDD Problems
just reading on the net and it would seem that your not alone, some one mentioned pulling the HDD out and attaching it to a pc and running some checks, they did, and they even sent it back to the manufacturer, here's a quote :
I sent the HDD back as Toshiba said it "could be the HDD", i replaced it with a Hitachi, I then tried to initialize it and it failed again !...i then changed image to new nabilo(what an image this is) and it initialized and works 100%, watched a couple of movies I'd transfered of the pc...
For every event that occurs,there will follow another event whose existence was caused by the 1st, and this second event will be pleasant or unpleasant according as its cause was skilful or un-skilful. ®
18th October 2011, 23:07 #3
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Re: USB and External HDD Problems
Ive allready tried 2 other images as well with same results, think it might be the path. Have tried editing the fstab file via dcc but Im not to sure about what Im doing there.
19th October 2011, 00:44 #4
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I had a prob like yours was a bit ago but I had to tellnet to dm800 USB/movie it was the rec path. Hope that helps it was 4/5 mths ago.
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PHOTOS: Crime Scene in the Bronx
Sources have confirmed that the dismembered body found scattered along several streets in Melrose is Tanya Byrd. The body parts were found stuffed into bags at around 4:30 a.m. in the area around Eagle and Cauldwell avenues, and East 156th and 158th streets.
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Instant Freeze Sculpting Mousse. Imagine yourself with the super-power to instantly freeze hair Indeed. Thanks to a foam that doesn't leave hair sticky. Aussie instant freeze hair spray, maximum. Aussie instant freeze hair spray, maximum gives you multiple styles. Which works out, because the plural of mousse is still mousse.
Frizzification got you feeling down under? No worries mate - Sydney Smooth's here. Learn the lingo and become one Sydney Smooth talker. Before you know it, you'll be saying: It's my way - or the highway - for you fly-aways, with confidence. Now hop to it! Contains natural Australian ingredients. Made in USA of US and/or imported ingredients.
Conditions to quench dry, thirsty hair. Moist Conditioner, with splashes of natural Aloe, replenishes natural moisture for healthy locks worth gushing about. Contains Natural Australian Ingredients Aussie Moist Conditioner conditions to quench dry, thirsty hair. Accented with natural Aloe, Moist Conditioner replenishes natural moisture for silky locks. Moist conditioner, with splashed of...
Cleans dull hair to reveal great shine. With touches of natural Guava and Sea Kelp, Sun-Touched Shine Shampoo reveals your natural shine in no time. Contains Natural Australian Ingredients Aussie Sun-touched Shine Shampoo let some light in! With touches of natural Guava and Sea Kelp, Sun-Touched Shine Shampoo reveals your natural shine in no time.
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Satisfy thirsty locks and add shine. Don't rinse. Do style.
Tame your mane with Aussie Men Texturizing Wax. Aussie Men Texturizing Wax: Hold level strong No worries