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· Quickstart · Docs · Comments

scssphp is a compiler for written in PHP.

SCSS is a CSS preprocessor that adds many features like variables, mixins, imports, color manipulation, functions, and tons of other powerful features.

The entire compiler comes in a single class file ready for including in any kind of project in addition to a command line tool for running the compiler from the terminal.

scssphp implements SCSS (3.2.12). It does not implement the SASS syntax, only the SCSS syntax.

Follow the author on twitter: @moonscript .

Installing

You can always download the latest version here: scssphp-0.0.12.tar.gz

You can also find the latest source online: https://github.com/leafo/scssphp/

If you use Tods Thea small shoulder bag Inexpensive Cheap Price dm4no
for installing packages, then you can update your like so:

Language Reference

For a complete guide to the syntax of SCSS, consult the Pointed Toe Slip On Flat Heel Shoes DEEP GREEN Sale Latest Collections Clearance Genuine For Cheap Discount Free Shipping Real PYj2CBc
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PHP Reference

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Quickstart

If you just want to start serving compiled files as quick as possible then start here.

scssphp comes with a easy to use class that automatically compiles modified files and serves them from a directory you specify.

Create a file, like :

Create the directory set in the script alongside the script, then add your files to it.

If we've got a file in there called , then we just need to hit the url: to get the compiled css.

If there is an error compiling, the url will result in a error with the error message. If the file can’t be found, then a friendly is returned.

scssphp will automatically create a directory inside the stylesheets directory where it will cache the compiled output. This way it can quickly serve the files if no modifications have been made. Your PHP script must have permission to write in .

Compiler Interface

If you're interested in directly using the compiler, then all you need to do is require and invoke the class:

The method takes as a string, and returns the . If there is an error when compiling then an exception is thrown with an appropriate message.

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Issues

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Changelog

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0.0.12
Figure 3

Aberrant distribution of apical proteins in FHL5. Laser-scanning immunofluorescence micrographs of duodenal enterocytes of patient 2 show () mislocalization of subapical Rab11 to the cytoplasm and accumulation around microvillus inclusions (white arrowheads) in contrast to controls and () mislocalization of apical Stx3 to the cytoplasm in association with microvillus inclusions (white arrowheads) in comparison to controls. ( and ) Brush-border microvilli are indicated by actin staining. Scale bar: 10 μm.

In conclusion, the ultrastructural characteristics of duodenal enterocytes from the FHL5 patients presented herein appeared identical to those from reported MVID patients ( 17 , Yves Salomon Army detachable collar bomber jacket Free Shipping Sale High Quality Cheap Sale Clearance Footlocker Cheap Online IKlUbRQM7v
). All major morphological hallmarks of MVID were found: first, loss or shortening of apical microvilli; second, a subapical accumulation of PAS-positive vesicles and tubules; and, finally, MI and/or basolateral microvilli.

Targeted disruption of Munc18 recapitulates intestinal features of FHL5. In order to analyze the molecular mechanism underlying MVID-like enteropathy associated with FHL5, we first generated a disease model cell line. For this, we used CaCo2 cells, a human enterocyte model cell line, which is able to establish enterocyte polarity in culture ( Jil Sander adjustable buckle trousers With Paypal Free Shipping Low Shipping Fee For Sale Discount Big Discount Cheap Latest Collections JFnOp71
). Using CRISPR/Cas9-mediated genome editing, we specifically targeted the STXBP2 locus. This resulted in a complete loss — KO — of Munc18 protein level ( Givenchy Eyewear Circle Wire sunglasses Outlet Locations Popular Marketable Shipping Outlet Store Online Free Shipping Clearance Store eHloi4D5nC
). To control for potential off-target effects by genome editing, we reintroduced a HA-Strep-TactinII–tagged (HS-tagged) version of WT Munc18 (HS-Munc18_gR) and the reported mutation/deletion of FHL5 patient 2 (HS-Munc18_gR-d232) in Munc18-KO cells ( Figure 4A ). The 232del mutant was exemplarily chosen for further mechanistic studies. Both cDNAs harbor additional noncoding mutations, rendering them resistant to the targeting sequence used for CRISPR/Cas9. Since loss of enterocyte polarity is a feature of intestinal FHL5, we subjected the model cell lines to cyst formation, an assay for challenging the establishment of epithelial polarity ( 26 ). Indeed, loss of Munc18 resulted in a significantly decreased single central lumen formation in comparison with the WT CaCo2 cells. This effect could be reverted upon the reintroduction of the HS-Munc18_gR. However, HS-Munc18_gR-d232 failed to revert the phenotype ( Wholesale Price Sale Online Cheap Fake Local Authority embroidered palm tree cap z36eN
).

Figure 4

Loss of Munc18 disrupts enterocyte polarity. () Western blot showing CaCo2 WT, Munc18-KO (gMunc18), and Munc18-KO cells reverted with HS-tagged, gRNA-resistant Munc18 WT and d232 mutant. () Quantification of single lumen formation (bounds of boxes represent 25th/75th percentiles, lines within boxes indicate medians, and whiskers represent minimum/maximum; statistical significance was tested by Student’s test; *** < 0.005, ≥ 100 cysts/experiment, 3 independent experiments). () Polarity is disrupted upon Munc18 depletion (gMunc18) in a 3D cyst formation assay in comparison to WT. Reexpression of HS-Munc18_gR reverts the phenotype, while HS-Munc18_gR-d232 does not. Actin is labeled in red an nuclei in blue; basal actin stain is seen in reaction to extracellular matrix (e.g., Matrigel). () Loss of Munc18 results in loss of apical microvilli (arrows) and subapical vesicle accumulation (arrowheads) in CaCo2 cells. Scale bar: 1 μm. () Reexpression of HS-Munc18_gR reverts the Munc18-KO phenotype. The brush-border microvilli are indicated with arrows. Scale bar: 10 μm (); 1 μm ().

To analyze the morphology in greater detail, WT, KO, and reverted cells were grown on porous membrane supports in order to establish enterocyte polarity. In KO cells, EM revealed a reduction and shortening of apical microvilli and an aberrant accumulation of subapical vesicles ( Figure 4D ). The detected ultrastructural aberrations were specific for the loss of Munc18, as the reverted cell line displayed a regular enterocyte ultrastructure ( Figure 4E ).

Fig. 2.

Rab27a regulates neutrophil migration by controlling uropod release. ( A ) Tracks of WT and Rab27a KO BM-PMN migration measured from stacks of images taken every 30 seconds for 30 minutes in a Zigmond chamber with medium alone added to the left well and 10 nM MIP-2 added to the right well. Black and red tracks indicate cells with net leftwards or rightwards movement respectively. Migration distance ( B ), velocity ( C ) - and -migration index ( D ) and uropod lifetime ( E ) of WT and Rab27a KO BM-PMN. Data represent the mean±s.e.m. from 25 random cells per experiment from four independent experiments (=4 for each genotype of BM-PMNs). Still images of WT ( F ) and Rab27a KO ( G ) BM-PMN migration over a course of 180 seconds (from supplementary material Movies 2 and 4). Scale bars: 10 μm. *≤0.05, ***≤0.001 (Student's -test).

In conclusion, we found that Rab27a regulates PMN chemotaxis in vitro and in vivo by regulating uropod detachment. We observed that Rab27a-positive membranes localised at the rear of migrating PMNs, suggesting that this protein has a role in cargo delivery to sites proximal to the uropod. Furthermore, we found that cell surface CD11b turnover was reduced in chemokine-stimulated Rab27a KO BM-PMNs and that Rab27a regulates the release of a subset of serine proteases. We propose that Rab27a regulates BM-PMN migration by stimulating elastase release from azurophilic granules at the rear of BM-PMNs, thereby allowing localised proteolysis of the extracellular domain of CD11b resulting in uropod detachment and net forwards movement of the cell. In view of recent data highlighting the role of Rab27a in T cell migration ( Colvin et al., 2010 ) and Rab27b in breast cancer cell invasion ( Kuboraum round shaped glasses Marketable ddjbBR0xgF
), we suggest that Rab27 might play a general role in regulating cell migration by allowing localised detachment at the rear of migrating cells.

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Mouse strains and purification of neutrophils

The ashen (Rab27a KO) and transgenic EGFP–Rab27a mice were as described previously ( Cheap Price Fake Lost amp; Found Rooms long printed sheer Tshrit Cheapest Sale Online AZg9i2LO
; Isabel Marant shearling belted jacket Where To Buy Cheap Real NzoxNaULQ
). Controls were age- and sex-matched C57BL/6 animals. All animals were treated humanely in accordance with the UK Home Office Regulations under PPL 70/7078. BM-PMNs were purified as described previously ( Cult Gaia Ark bag The Cheapest For Sale Outlet Official Site Clearance Amazing Price Best Store To Get Cheap Online Marketable BChcsf
). Typical preparations contained above 80% BM-PMNs.

Antibodies, immunoblot and flow cytometry

For immunoblot analysis, mouse monoclonal anti-Rab27a antibody (4B12) ( Marcelo Burlon County Of Milan Snake Wing Tshirt Buy Cheap Low Price Fee Shipping gg5lXtg
), anti-calnexin antibody (Stressgen) and HRP-conjugated secondary antibodies (DAKO) were used as described previously ( Barral et al., 2002 ). Cells were lysed by resuspension in 50 mM Tris-HCl pH 7.4, 1× protease inhibitors (Roche), 2% (w/v) SDS, and passed through a 26G needle 10 times on ice and processed for immunoblotting as previously described ( Barral et al., 2002 ). For flow cytometry using a FACScalibur cytometer, phycoerythrin (PE)-conjugated anti-Ly6G (BD Biosciences), allophycocyanin (APC)-conjugated CXCR2 (RD Systems), Alexa-Fluor-647-conjugated anti-Cd11b (BD Biosciences) antibodies were used at 1:100 dilution and Alexa-Fluor-647-conjugated phalloidin (Molecular Probes) used at 1:1000 dilution. For surface staining, 2×10 BM-PMN cells were washed twice and resuspended in rat anti-FcγRII/RIII antibody (BD biosciences), then incubated with fluorophore-conjugated antibodies for 30 minutes on ice. For F-actin staining, 3×10 BM-PMN cells were fixed for 30 minutes at room temperature in 4% paraformaldehyde (PFA) and permeabilised with 0.5% saponin for 15 minutes prior to Alexa-Fluor-647–phalloidin staining. For Cd11b measurements, 2×10 BM-PMNs were resuspended in RPMI1640 plus 1% BSA, stimulated with 1 nM MIP-2 at 37°C for indicated times and placed on ice prior to Cd11b staining.

The human promyelocytic leukaemia cell line HL-60 (American Type Culture Collection) was cultured in DMEM, 20% (v/v) fetal bovine serum and 10 units/ml penicillin-streptomycin at 37°C under 5% CO 2 . HL-60 cells were nucleofected with non-targeting siRNA (Dharmacon) or two Rab27a-specific siRNA oligonucleotides using the Amaxa nucleoporator (Lonza) as described previously ( Munafó et al., 2007 ). Nucleofected cells were then differentiated to mature neutrophils by suspension in medium containing 1.3% DMSO for 72 hours.

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News The Essential Daily Briefing

The presenter contracted the illness on the way back to the UK

Victoria Sanusi 3 days
The i newsletter
News for open-minded people.

Television and radio presenter Richard Bacon has reportedly been placedin an induced coma while suffering from a respiratory virus.

The 42-year-old has been in a London hospital since last Thursday, when he arrived back from the US.

The former Blue Peter host, who is now based in Los Angeles, wasinitially diagnosed withpneumonia across both lungs, The Sun reported.

“Thank God I got ill in Britain (actually on the way to Britain, was taken off the plane in a wheelchair),” he tweeted last week.

“Fuck all the ideology driven politicians who’ve messed up America’s healthcare system. Viva the NHS. Happy 70th,” he added, referring tothe health service’s landmark anniversary.

Thank God I got ill in Britain (actually on the way to Britain, was taken off the plane in a wheelchair). Fuck all the ideology driven politicians who’ve messed up America’s healthcare system. Viva the NHS. Happy 70th. pic.twitter.com/ABEhSZXiRK

— richard bacon (@richardpbacon) July 5, 2018

According to Red Valentino floral threequarter sleeve sweater Cheap Sast For Sale Cheap Price From China Discount Find Great dkghTC
,Bacon replied to afan who quizzed him on his condition, he explained: “[It’s] basically pneumonia.

“But across two lungs.Although I thought that was for people in their 80s, I’ll be here a few days.”

The news comes after the TV star revealed that he was diagnosed with ADHD in his forties.

Speaking to This Morning , he said “I was causing chaos for my family… For my wife in particular… It all adds up now,

“In many ways I was pleased to get the diagnosis,

“Aphysiatrist looked at the brain scan and said ‘You are lucky to have made it to 42. People like you are dangerous. You are impulsive’ and he gestured to [my wife] Rebecca and he said: ‘This woman is saving your life'”.

Read more:

“I was terrified, pleading with God.” This is what a near death experience feels like

Man recovering from coma after brain ‘jump-started’ using ultrasound

Darius Campbell went into a life-threatening coma after drinking dirty Thames water

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