Genetic manipulation of emerging bacterial pathogens, such as coagulase-negative staphylococci (CoNS), is a major hurdle in clinical and basic microbiological research. Strong genetic barriers, such as restriction modification systems or clustered regularly interspaced short palindromic repeats (CRISPR), usually interfere with available techniques for DNA transformation and therefore complicate manipulation of CoNS or render it impossible. Thus, current knowledge of pathogenicity and virulence determinants of CoNS is very limited. Here, a rapid, efficient, and highly reliable technique is presented to transfer plasmid DNA essential for genetic engineering to important CoNS pathogens from a unique Staphylococcus aureus strain via a specific S. aureus bacteriophage, Φ187. Even strains refractory to electroporation can be transduced by this technique once donor and recipient strains share similar Φ187 receptor properties. As a proof of principle, this technique was used to delete the alternative transcription factor sigma B (SigB) via allelic replacement in nasal and clinical Staphylococcus epidermidis isolates at high efficiencies. The described approach will allow the genetic manipulation of a wide range of CoNS pathogens and might inspire research activities to manipulate other important pathogens in a similar fashion.
Coagulase-negative staphylococci (CoNS), such as human skin-colonizing Staphylococcus epidermidis or Staphylococcus lugdunensis, are frequently isolated from hospital-associated infections and represent emerging bacterial pathogens (1, 2). Many CoNS, in particular hospital-associated S. epidermidis, are resistant to available antibiotics, such as methicillin (≥75% of S. epidermidis isolates), clindamycin, tetracycline, trimethoprim, macrolides, and aminoglycosides (3). Such strains typically cause a variety of complicated infections, particularly infections associated with indwelling medical devices in combination with strong biofilm formation or, even more alarming, life-threatening systemic disease, such as endocarditis or sepsis (2, 4–6).
Genetic engineering of the well-studied pathogen Staphylococcus aureus has revolutionized research activities in the field of staphylococci in the past. However, current knowledge of the physiology and pathogenicity of CoNS is very limited because most available techniques used for genetic manipulation (e.g., electroporation of shuttle plasmids) often fail, most likely because of strong genetic barrier mechanisms, such as restriction-modification (R-M) systems or clustered regularly interspaced short palindromic repeats (CRISPR), previously shown to impede horizontal gene transfer (HGT) events between bacteria (7, 8). However, specifically engineered Escherichia coli strains lacking the dcm gene required for cytosine methylation of DNA have been developed to produce plasmid DNA that is not degraded upon electroporation of S. aureus or of specific S. lugdunensis strains, or even of the clinical S. epidermidis isolate RP62A (9, 10). This approach is promising but requires large amounts of plasmid DNA (typically 5 μg) and seems to be limited to specific CoNS isolates. An alternative to electroporation is protoplast transformation, which is very ineffective in the case of S. epidermidis because of its natural insensitivity to lysostaphin, an enzyme used to degrade staphylococcal cell walls (11). Equally problematic is transduction. Although transducing S. epidermidis phages have been described in the past (12–14), they are suitable solely for DNA transfer among S. epidermidis isolates, but not for DNA introduction from genetically more amenable organisms, such as S. aureus. For such approaches, interspecies transduction events are required, which have been reported, for example, between S. aureus and S. epidermidis isolates via S. aureus phage Φ80 (15). However, because most S. aureus DNA donor strains synthesize ribitol-phosphate wall teichoic acid (WTA) phage receptors distinct from those of most CoNS recipients, phage adsorption of S. aureus transducing phages, such as Φ80 or Φ11, to CoNS is usually blocked, rendering HGT impossible (16–19). Thus, the available techniques are highly challenging and require multiple attempts or, as is more often the case, completely fail with clinical CoNS, which represents a major hurdle for the molecular characterization of CoNS.
Recently, phage-mediated HGT of S. aureus pathogenicity islands (SaPIs) between major bacterial pathogens has been reported (17). Surprisingly, the S. aureus ST395 lineage-specific phage Φ187 has been capable of transferring SaPIs between ST395 isolates and many CoNS because of shared properties in the WTA cell surface receptors (17). Genomic and biochemical analyses of the ST395 WTA biosynthesis pathway have further suggested the occurrence of previous HGT events between ST395 isolates and CoNS, most likely via Φ187-related phages (20). Because most pathogenic CoNS, such as S. epidermidis, have a glycerol-phosphate WTA backbone resembling that of ST395 isolates, Φ187 might be a suitable tool to transfer plasmid DNA to CoNS strains that are otherwise difficult to transform.
Here, a highly efficient method to introduce plasmid DNA into important CoNS pathogens for genetic manipulation is described. The method aims to transfer plasmid DNA via phage Φ187 from a genetically engineered ST395 isolate to major clinical CoNS isolates, and even to strains refractory to electroporation protocols, for their genetic engineering.
Bacterial strains and growth media.All the bacterial strains listed in Table 1 were grown at permissive temperatures in basic medium (BM) (1% tryptone, 0.5% yeast extract, 0.5% NaCl, 0.1% K2HPO4, 0.1% glucose) or in lysogeny broth (Becton Dickinson) supplemented with appropriate antibiotics (10 μg/ml for chloramphenicol, 12.5 μg/ml for tetracycline, or 100 μg/ml for ampicillin). For experiments performed on solid medium, BM agar plates containing 5% sheep blood were used unless otherwise noted.
Molecular genetic methods.Plasmid transformation was performed according to the method of Winstel et al. (17). Briefly, plasmids were isolated from appropriate E. coli, Staphylococcus carnosus TM300, S. aureus RN4220, or S. aureus PS187 ΔhsdR ΔsauUSI donor strains by using standard techniques, purified, and electroporated into electrocompetent recipient strains using a 1-mm-gap electroporation cuvette. The cells were pulsed at 1,000 V, and subsequently, 950 μl prewarmed BM (37°C) was added. Bacteria were grown for 70 min at 37°C, plated onto selective medium, and grown overnight at 37°C (or 30°C for knockout plasmids). Transformants were counted to calculate the transformation efficiency.
For the construction of ermB-bearing S. epidermidis sigB mutants, an ermB resistance cassette was amplified from plasmid pEC2 (21) with primers lox66erm and lox71erm (Table 2) and ligated with the SmaI-digested pKOR1-like knockout plasmid pBASE6 (22), resulting in plasmid pBASE6-erm/lox2. The flanking regions of sigB were amplified from genomic DNA of S. epidermidis TÜ3298 with primer pairs sigflank1 and sigflank2 (digested with BspEI and Acc65I) (Table 2) and sigflank3 and sigflank4 (digested with NheI and SalI) (Table 2), respectively. The digested flanking regions were consecutively ligated with the identically digested knockout plasmid, resulting in pBASE6-sigB-ko2, which was used to transform E. coli DC10B (9) and S. aureus PS187 ΔhsdR ΔsauUSI cells.
Experiments with phage.To propagate Φ187, its cognate host strain S. aureus PS187 wild type (w.t.) was grown overnight in BM and diluted in Φ187-containing lysates (1 × 109 PFU/ml; titrated on PS187 wild type as described previously ) to a final optical density at 600 nm (OD600) of 0.4. After the addition of CaCl2 (final concentration, 4 mM), the bacteria-phage suspension was incubated without shaking at 37°C for 30 min and subsequently for at least 3 h at 30°C with slight agitation until complete lysis occurred. Cell debris was pelleted via centrifugation (10 min; 5,000 × g), and phage-containing supernatants were filter sterilized (0.02 μm) and stored at 4°C or subsequently used to infect overnight cultures of pKOR1-, pRB474-, or pTX15-bearing S. aureus PS187 ΔhsdR ΔsauUSI grown in BM (obtained via electroporation [see above]). The infection parameters were as described before, except that bacteria were diluted in phage lysates to a final OD600 of 1.0 and incubated at 30°C, which usually required at least 5 h for lysis to occur. Plasmid-bearing phage particle (PBPP) lysates were clarified via centrifugation (10 min; 5,000 × g), filter sterilized, and titrated on PS187 wild type.
For plasmid transfer to CoNS, the bacterial densities of overnight cultures of plasmid recipient strains (e.g., S. epidermidis TÜ3298) were adjusted to an OD600 of 0.5 in BM. Approximately 8.0 × 107 bacteria were centrifuged and resuspended in 200 μl phage buffer (4 mM CaCl2, 1 mM MgSO4, 50 mM Tris-HCl, pH 7.8, 100 mM NaCl, 0.1% gelatin), mixed with 100 μl PBPP lysate (∼1 × 109 PFU/ml), and incubated at 37°C for 15 min with slight agitation, allowing phage particle adsorption to bacterial host cells. Afterward, the bacteria-phage suspension was plated onto selective media containing appropriate antibiotics and incubated for 24 h at 37°C (or 48 h at 30°C for knockout plasmids). The resulting transductants were counted to calculate transduction efficiencies, which are given as transductants per milliliter of phage lysate.
Phage Φ187 adsorption efficiencies were determined as described previously with minor modifications (17). Briefly, adsorption rates were analyzed using a multiplicity of infection (MOI) of 0.1, and the adsorption rate (percent) was calculated by determining the number of unbound PFU in the supernatant and subtracting it from the total number of input PFU as a ratio to the total number of input PFU.
Ultracentrifugation of PBPP lysates.For enrichment of PBPP, lysates were ultracentrifuged for 2 h at 25,100 rpm (73,000 × g) at 4°C using a Beckman 45Ti ultracentrifuge rotor. Subsequently, the PBPP-containing pellet was resuspended in an appropriate amount of TMN buffer (10 mM Tris-HCl, pH 7.5, 10 mM MgSO4, 500 mM NaCl), and the lysates were titrated on PS187 wild type and used for plasmid transfer experiments, as previously described.
Tests for natural competence and "pseudotransformation."To analyze the natural competence of selected S. epidermidis isolates, PBPP lysates (containing pTX15) were treated with 20 U DNase I for 1 h at 37°C to digest extracellular DNA and subsequently used for plasmid transfer to strain TÜ3298, 101 b, or 204/70131537, as previously described. In a second approach, and to inactivate phage particles but not DNA, PBPP lysates (containing pTX15) were heat inactivated for 10 min at 70°C, chilled, and subsequently used for plasmid transfer to selected S. epidermidis strains. Controls were not heat inactivated or lacked DNase I treatment. To determine phage-mediated "pseudotransformation," selected S. epidermidis strains were incubated with 1.0 μg purified pTX15 plasmid in the presence or absence of Φ187 w.t. particles (∼1 × 109 PFU/ml) for 15 min at 37°C. Φ187 w.t. particles were previously propagated on S. aureus PS187 w.t., which lacks pTX15, pRB474, and pKOR1 plasmids. The bacteria-phage suspension was plated onto selective medium containing the appropriate antibiotic and incubated for 24 h at 37°C. Transductants were counted to calculate the transduction efficiency, as previously described.
Allelic replacement.Allelic replacement was performed as previously described with minor modifications (23). Briefly, S. aureus PS187 ΔhsdR ΔsauUSI bearing a sigB knockout plasmid was infected with Φ187 to create a PBPP lysate, as previously described, which was subsequently used for plasmid transfer to S. epidermidis. The knockout plasmid was integrated into the genome of the target strain in the presence of chloramphenicol (10 μg/ml) at a temperature of 42°C. Following plasmid integration, two consecutive counterselection steps using anhydrotetracycline at concentrations of 0.2 μg/ml and 1.0 μg/ml were performed. The resulting colonies were transferred to BM agar plates with or without chloramphenicol (10 μg/ml). Chloramphenicol-sensitive colonies were used for PCR analysis to confirm the sigB deletion.
Statistical analysis.Statistics were performed using GraphPad Prism (version 5.04; GraphPad Software, Inc., La Jolla, CA, USA). Statistically significant differences were calculated using the unpaired two-tailed Student t test.
Inactivation of restriction systems converts S. aureus strain PS187 into a suitable intermediary host for plasmid DNA.S. aureus ST395 isolate PS187 accepts S. aureus RN4220-derived plasmid DNA at low efficiencies, but inactivation of the restriction systems HsdR and SauUSI has rendered the strain highly susceptible to plasmid transformation (17). Lack of both restriction systems may also enable direct electroporation of strain PS187 with plasmid DNA from sources other than S. aureus. To this end, various plasmids widely used for targeted mutagenesis (pKOR1 ), overexpression (pTX15 ), or complementation (pRB474 ) studies were isolated from either E. coli or S. carnosus using standard techniques and introduced into S. aureus PS187 wild type and its restriction-deficient mutant PS187 ΔhsdR ΔsauUSI via electroporation. PS187 wild type was resistant to electroporation with DNA derived from E. coli, while deletion of restriction systems resulted in robust transformation efficiency (Fig. 1). PS187 wild type could also be transformed at low efficiency with plasmid DNA derived from S. carnosus TM300, although the absence of restriction systems significantly increased the electroporation frequency. Thus, S. aureus PS187 ΔhsdR ΔsauUSI is easily transformable and represents a suitable alternative to the S. aureus strain RN4220 conventionally used as an intermediary host for plasmid DNA.
Lack of restriction systems converts S. aureus strain PS187 into a suitable intermediary host for plasmid DNA. Shown are the electroporation frequencies of S. aureus PS187 wild type (w.t.) and its restriction system-deficient mutant PS187 ΔhsdR ΔsauUSI. Cells were electroporated with purified E. coli DH5α-derived pRB474, E. coli DB3.1-derived pKOR1, or S. carnosus TM300-derived pTX15 plasmid. The values represent transformants per microgram of DNA and are given as means and standard deviations (SD) (n = 3). Statistically significant differences calculated by the unpaired two-tailed Student t test are indicated: **, P < 0.001 to 0.01; ****, P < 0.0001.
Most CoNS are refractory to common electroporation protocols.In addition to alterations of WTA structure that diminish phage infection and CRISPR loci that degrade foreign DNA, R-M mechanisms involving distinct DNA methylation patterns represent significant barriers to HGT. The fact that ST395 isolates are capable of exchanging DNA with certain CoNS has led to the assumption that ST395 and CoNS share similar DNA methylation pathways, enabling phage-mediated HGT between donors and recipients with shared WTA structures (17). Therefore, S. aureus PS187 ΔhsdR ΔsauUSI-derived plasmid DNA (or RN4220-derived plasmid DNA, used as a control) might be suitable for electroporation of CoNS. When plasmids pRB474 and pTX15 from one of the two strains were used to electroporate various CoNS isolates, the majority of test strains could hardly be transformed, and some were not even detectably transformable under the electroporation conditions used in this study (Fig. 2A and B). Thus, most CoNS are refractory to electroporation even if plasmid DNA is derived from ST395 clone PS187.
Benefits of Φ187-mediated plasmid transfer to important CoNS pathogens. Shown is a comparison of plasmid electroporation (EP) (hatched bars) and Φ187-mediated plasmid transfer (transduction [TD]) in various S. epidermidis (A) and S. lugdunensis (B) strains. Purified pTX15 (gray bars) or pRB474 (black bars) plasmids were isolated from S. aureus strain RN4220 or PS187 ΔhsdR ΔsauUSI and used to electroporate electrocompetent CoNS (EP+ indicates application of 2.5 μg DNA). For transduction experiments, PBPP lysates contained ∼1 × 109 PFU/ml. The lysates were titrated on S. aureus PS187 w.t. The values represent transductants per milliliter of phage lysate (for TD experiments; plated on sheep blood-supplemented agar plates) or transformants per microgram of DNA (for EP experiments) and are given as means and SD (n = 3). Statistically significant differences calculated by the unpaired two-tailed Student t test are indicated: ns, not significant (P > 0.05); *, P < 0.01 to < 0.05; **, P < 0.001 to 0.01; ***, P < 0.001; ****, P < 0.0001.
S. aureus phage Φ187 facilitates plasmid transfer to untransformable CoNS via transduction.As an alternative to electroporation, phage-mediated transduction of DNA can be used to manipulate bacteria. The ability of the ST395-specific S. aureus phage Φ187 to transfer SaPIs from ST395 to CoNS suggested the use of Φ187 as a molecular shuttle for plasmid transfer to CoNS. Therefore, Φ187 was freshly propagated on its cognate host strain S. aureus PS187 wild type and used to infect plasmid-bearing S. aureus PS187 ΔhsdR ΔsauUSI to create PBPP lysates, which were used to infect a variety of CoNS strains. Surprisingly, the novel approach facilitated highly efficient plasmid transfer at frequencies up to ∼104 transductants per ml of PBPP lysate to many CoNS and even to strains usually refractory to electroporation (Fig. 2A and B). Moreover, enrichment of PBPP lysates via ultracentrifugation resulted in both higher phage titers (up to 1 × 1011 PFU/ml) and more efficient plasmid transfer frequencies (see Fig. S1A in the supplemental material). Although plasmid transfer with ultracentrifuged PBPP lysates is recommended, titers of 109 PFU/ml were usually sufficient to accomplish plasmid transfer. Interestingly, the use of BM agar plates containing 5% sheep blood increased the plasmid transfer efficiency (see Fig. S1B in the supplemental material). This effect might be a consequence of supplements other than CaCl2 present in whole blood that facilitate more efficient phage adsorption and infection.
To confirm that plasmid transfer was indeed accomplished via Φ187-mediated transduction and not by a phage-independent mechanism involving, for example, uptake of free DNA (a process that is very rare among staphylococci but has recently been documented in S. aureus under specific conditions ), PBPP lysates were treated with DNase I. While DNase I treatment of PBPP lysates did not alter the efficiency of plasmid transfer to different S. epidermidis strains (a test for natural competence), heat inactivation of phage particles in PBPP lysates rendered plasmid transfer undetectable (Fig. 3A and B). Plasmid transfer to CoNS failed even when naked plasmid DNA (pTX15) lacking phage particles or naked DNA was combined with Φ187 particles propagated on PS187 wild type without plasmids (a test for "pseudotransformation" ), thereby confirming that plasmid transfer was accomplished via Φ187, which is in line with a transduction process (Fig. 3C). Thus, Φ187 is capable of transducing plasmid DNA from ST395 isolates to CoNS, even those strains that are usually refractory to electroporation.
Plasmid transfer to CoNS is facilitated by Φ187 via transduction. (A) Analysis of natural competence of selected S. epidermidis isolates. PBPP lysates (containing pTX15) were treated with DNase I (+) or untreated (−) and subsequently used for plasmid transfer to strain TÜ3298, 101 b, or 204/70131537. (B) Effect of heat treatment of PBPP lysates (containing pTX15). PBPP lysates were heat inactivated (+) or not (−), chilled, and subsequently used for plasmid transfer to selected S. epidermidis strains. (C) Analysis of selected S. epidermidis isolates to undergo "pseudotransformation." Purified pTX15 plasmid was incubated with selected S. epidermidis strains in the presence (+) or in the absence (−) of Φ187 wild-type particles lacking plasmids. The lysates contained ∼1 × 109 PFU/ml. The values represent transductants per milliliter of phage lysate plated on sheep blood-supplemented agar plates and are given as means and SD (n = 3). Statistically significant differences calculated by the unpaired two-tailed Student t test are indicated: ns, not significant.
Clinically relevant CoNS pathogens can be transduced via phage Φ187, facilitating their genetic manipulation.The new approach was tested with a broad panel of CoNS, including S. epidermidis and S. lugdunensis clinical and nasal isolates (details are provided in Table 1). Notably, the majority of S. epidermidis (65.2%) and S. lugdunesis (75.0%) strains could be transduced using various Φ187-based PBPP lysates (Fig. 4A and B). While some S. epidermidis (e.g., TÜ3298) or S. lugdunensis (e.g., N920143) strains underwent highly efficient Φ187-mediated plasmid transfer, some isolates (e.g., S. epidermidis ATCC 14990 or S. lugdunensis HKU09-01) could be transduced only with certain plasmids or were completely resistant, most likely because of genetic barriers inactivating the invading plasmid DNA. However, high-frequency plasmid transfer clearly correlated with strong Φ187 adsorption or, in contrast, failed when no or weak phage adsorption occurred (see Fig. S2A and B in the supplemental material). In agreement with these observations, Φ187 adsorption to clinical Staphylococcus caprae isolates and Staphylococcus simulans correlated with transduceability. In contrast, non-Φ187-binding species, such as Staphylococcus saprophyticus, Staphylococcus warneri, or Staphylococcus haemolyticus, could not be transduced by Φ187, suggesting that similar cell surface receptor properties in the DNA donor and recipient are necessary for Φ187-mediated plasmid transfer to CoNS (Fig. 4C; see Fig. S2C in the supplemental material).
S. aureus phage Φ187 facilitates plasmid transfer to many untransformable staphylococcal pathogens. Bacteriophage Φ187 propagated on S. aureus donor strain PS187 ΔhsdR ΔsauUSI bearing plasmid pRB474 (black), pTX15 (gray), or pKOR1 (red) was used to transduce a broad panel of S. epidermidis (A) and S. lugdunensis (B) strains and other CoNS (C). The PBPP lysates contained ∼1 × 109 PFU/ml. The values represent transductants per ml of phage lysate plated on sheep blood-supplemented agar plates and are given as means and SD (n = 3). No transductants were observed in controls lacking PBPP lysates, and naturally antibiotic-resistant clones are indicated (resistant).
Finally, a plasmid for targeted mutagenesis was constructed and transduced to clinical and nasal S. epidermidis isolates using this approach. The gene encoding the alternative transcription factor sigma B (sigB) was used as a test gene. Notably, rapid gene deletion was achieved at high efficiency (Table 3). Thus, Φ187 is a suitable tool for transferring plasmids to important clinical CoNS pathogens and represents a novel tool for their genetic manipulation.
In summary, the new approach represents a major technical advance for plasmid transfer to important CoNS pathogens, allowing their genetic manipulation in the future. Although prior work from 1965 reported interspecies plasmid transfer events between S. aureus and S. epidermidis (15), the phenomenon seems to be limited to a small minority of S. epidermidis isolates and has hardly been used since. In contrast, the majority of important clinical CoNS isolates, including strong biofilm-producing S. epidermidis strains 1457 and RP62A and, equally important, genome-sequenced strains, can be transduced by the approach presented here in a highly efficient and reliable fashion. In addition, the transfer of knockout plasmids via Φ187 will strongly facilitate the genetic manipulation of CoNS in the future. Additionally, Φ187 might be capable of transducing chromosomal markers from ST395 to CoNS in conserved genomic regions shared by ST395 and CoNS to facilitate rapid gene disruption in CoNS. Although some CoNS isolates and species could not be transduced, most likely because of distinct phage receptor properties, lack of plasmid replication in the recipient strains, or other genetic barriers limiting HGT, the technique described here is applicable to most important and clinically relevant CoNS, including S. epidermidis and S. lugdunensis. This approach may prompt research activities to manipulate other important bacterial pathogens in a similar fashion.
We thank Karsten Becker and Evgeny Idelevich for clinical S. epidermidis strains.
This work was supported by German Research Council grants TRR34 to A.P. and Ro2413/4-1 to H.R. and by German Center for Infection Research (DZIF) grants to H.R. and A.P.
Accepted manuscript posted online 23 January 2015.
Supplemental material for this article may be found at http://dx.doi.org/10.1128/AEM.04190-14.
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I flung open the fridge and stuffed as many bottles of wine into my makeshift bag as it could fit. Now I just needed to get to the stall before they called for backup. I looked around the room for something to fend off the assailant as she raised a device to her mouth.
I grabbed the empty knife block and chucked it at her head, knocking her out cold. I picked up my bag and made my way to the stall. I could already hear the patter of backup arriving at the storage room as I entered the bathroom and locked the door.
I was already onto my third bottle by the time the door started pounding. I had lined the stall with as much aluminum foil as I could, but I knew it wouldn't hold. I could only hope that I'd black out before a dangerous memory returned.
My vision was getting blurry and I couldn't stand, but they were almost through the door. I managed to gulp down one last mouthful before the door fell and they stormed the room. Five armed guards apprehended me while a woman watched from the doorway. I tried to speak, but the darkness swallowed up my vision.
Following a patient's third episode this month, staff have been reminded that under no circumstance are they to interact with a patient after curfew without the presence of two armed guards. Additional cameras have been installed in all staff rooms and corridors, and at least one security personnel will be monitoring them at all times. We thank you for your service.


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text:
Eqbaliyeh (, also Romanized as Eqbālīyeh) is a village in Tankaman Rural District, Tankaman District, Nazarabad County, Alborz Province, Iran. At the time of the 2006 census, its population consisted of 494 individuals in 123 families.

References

Populated places in Nazarabad County

redpajama_set_name: RedPajamaWikipedia


Go to the best omakase spot in San Francisco and ask the sushi chef where you should go next - anywhere else. His recommendation? Buy a fishing rod, drive two hours up the coast, and wait. A small man will tap you on the shoulder. Follow him. That's all you need to know for now.
Just kidding. He'll actually just tell you to head west to Sushi Wako.
The tasting menu at Wako, on vastly underrated Clement Street, is pretty much a copy of the holy-grail omakase at Kusakabe. It's even a little cheaper.
But if you're dead set on having a sushi chef decide exactly what you're going to eat, and you're willing to spend as much as the $75 (roughly 12 avocado toasts) for that experience, you might as well spend the extra $20 (4 single-origin drip coffees) for Kusakabe. It's just a peg higher in our books.
That said, if you want to steer your own sushi ship, head to Wako. Unlike Kusakabe's all curated all the time setup, the S.S. Wako lets you order a la carte, and they also do a nigiri-only omakase in addition to a full tasting menu.
Wako has a neighborhood feel, but local cred only goes so far - Inner Richmond neighbors get turned away along with everyone else, because the 20-seat restaurant is very small. Seriously, it's no bigger than a San Francisco living room - not even the one you converted to another bedroom.
If you're coming to Wako, skip the full tasting menu and go for either for the omakase nigiri or order a la carte - we've got some recommendations on how to do so below. When you're done, ask the chef at Wako where he goes out for sushi. If it involves a fishing rod, good luck.
Bluefin Tuna from Spain and Nagasake Butterfish - both raw cuts are insanely tender. And don't skimp on the house soy sauce.
Cherry Blossom shrimp in japanese sweet potato pancake - a latke, as the chosen people call it, with bits of shrimp. Not kosher. Still amazing. Get this.
The chef's choice nigiri is the way to go if you want sushi. A few favorites: kelp-marinated Hokkaido halibut, the Spanish mackerel with ginger miso paste, the Stripe Jack, and Chinook king salmon from Canada. You don't have to remember all that, just eat what you're given and leave happy.
House panna cotta. We know sushi chefs have knife skills, but you have to see how tiny they chop the kiwi and strawberry on this. Mind boggling mincing.


Raytheon Space and Airborne Systems has reduced its CMMI SCAMPI Class A appraisal costs by over 43% and diminished program disruption while ensuring consistency of high maturity implementation. This presentation describes how Raytheon SAS, using an approach pioneered by Business Transformation Institute (BTI), achieved these results with no loss of appraisal fidelity and increased high maturity consistency. The presentation contrasts the traditional appraisal approach with integrating many small, near-real-time, incremental appraisals into the organization.
"Traditional" SCAMPI Class A appraisals, while valuable, are expensive, time-consuming, and disrupt participating programs. The hidden and greatest impact is in the programs as they gather artifacts and prepare for interviews.
Raytheon SAS' and BTI's new appraisal approach emphasizes small appraisals across all programs (not a select few) with minimal pre-appraisal artifact collection, replacing travel with web-based and telecom interactions, limited-duration interviews covering both artifact review and affirmations, and planning to promote high information re-use across process areas during appraisals.
The engine driving each appraisal is the "shoulder-to-shoulder" interview. During this interview, the appraisers ask questions and require program personnel to display artifacts in real-time. The team captures a record of artifacts shown and responses made. Implementing this approach requires deep CMMI knowledge, good communications skills, and excellent time management.
The result of the incremental appraisal approach was a 43% reduction in costs for Raytheon SAS' 2014 versus its 2011 appraisal, an even greater reduction in program impact, a more timely, consistent, and constant information flow to leadership, and better process understanding maturity integration into programs.
Raytheon Space and Airborne Systems (Raytheon SAS) is a 14,500+ person organization with multiple facilities located in Southern and Northern California, North Texas, Salt Lake City, Utah, Forest, Mississippi, Fort Wayne, Indiana, Southern Florida, Maryland, and Massachusetts. Raytheon SAS is headquartered in McKinney, Texas and builds radars and other sensors for aircraft, spacecraft, and ships. The business also provides communications and electronic warfare solutions and performs research in areas ranging from linguistics to quantum computing. Raytheon SAS serves the defense, space, and civilian sectors of the United States of America and a wide variety of international customers.
Raytheon SAS' appraisal approach for 2011 and prior years emphasized minimizing the risk of an unsuccessful appraisal. In this traditional appraisal approach, the organization would conduct one (or more) Class B appraisals as a dress rehearsal for the "big event". The activities culminated in an on-the-official record "big bang" Class A appraisal. As expensive as the appraisal event is, the hidden and greatest impact is on the programs as they gather artifacts and prepare for interviews.
In 2012, Raytheon SAS decided to radically change its appraisal approach in order to reduce program impact and appraisal costs while focusing on increasing high maturity consistency. Raytheon SAS teamed with BTI to achieve these goals and, in August 2014, Raytheon SAS successfully renewed its CMMI-DEV version 1.3 Maturity Level 5 credential for Raytheon SAS Engineering systems, hardware, and software product development programs that include new development using this new approach. An approach which cut previous appraisal costs by 43%.
Raytheon SAS' and BTI's new appraisal approach emphasizes small appraisals across all programs (not a select few) with minimal pre-appraisal artifact collection, replacing travel with web-based and telecom interactions, limited-duration interviews covering both artifact review and affirmations, planning to promote high information re-use across process areas during appraisals, and efficient data transfer from appraisal workbooks into preliminary and final findings presentations.
Raytheon SAS began conducting frequent, very small Class B appraisals in 2012. This sequence of low-cost and low-impact appraisals culminated in a SCAMPI Class A appraisal in August 2014 conducted by Business Transformation Institute, Inc. This appraisal results in Raytheon SAS continuing as a Maturity Level 5 organization for systems, software, and hardware engineering.
Raytheon SAS' performance improvement initiatives, of which the CMMI has been an integral part, have produced a significant reduction in schedule (SPI) variability, a significant improvement in defect containment, increased cost performance (CPI) and schedule performance (SPI) across the organization, increased software and hardware productivity, and a significant reduction in engineering rework. Because of the improvements seen in these areas CMMI will continue to be a part of SAS' improvement initiatives.
Raytheon SAS depends on the engineering innovation, efficiency, and fidelity of its processes to provide effective and affordable solutions to its customers. Improving processes is one of the key ways that Raytheon SAS integrates new ideas, better return-on-investment, and diminished rework across the organization and into programs.
Given the ever more cost-constrained marketplace in which Raytheon SAS operates and the cost of conducting a traditional SCAMPI Class A appraisal, Raytheon SAS is always sensitive to the cost of an appraisal versus the benefits derived from that appraisal. Raytheon SAS' new incremental appraisal approach has helped adapt a process improvement and measurement tool (CMMI and SCAMPI appraisals) that Raytheon SAS has found valuable in the past in this cost-constrained environment. As a result, Raytheon SAS leadership is propagating Raytheon SAS' CMMI approach into its future CMMI roadmap, across more of the organization and into other recently acquired organizations, while at the same time.


Senator Jim Oberweis is calling on Illinois consumers to help him in his push to end the state's Sunday sales ban for car dealers.
The 25th District Senator has introduced Senate Bill 1780 to repeal the 32-year prohibition that has prevented Illinois consumers from being able to buy cars on Sundays. He is also sponsoring Senate Bill 1706 to allow for the sale of motor vehicles by licensed dealers on Sundays if the dealer is a person who observes a religious day of worship other than Sunday; and Senate Bill 1835 to allow for the sale of motor vehicles by licensed dealers for no more than two hours on Sundays.
All three bills were allowed a "subject-matter-only" committee hearing by the Senate Transportation Committee April 21, but no vote was taken. Testifying on behalf of those bills were Heather Niemetschek of the Illinois Policy Institute (pictured at right) and Dyanne Ferk of the University of Illinois (pictured at left).


text:
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Hyundai Sonata N-Line coming with over 275-hp
Yesterday Hyundai officially unveiled the 2020 Hyundai Sonata at the New York Auto Show. The 2020 Sonata is a bold new midsize...
Hyundai will demonstrate its new Digital Key in New York
Next week Hyundai has some big debuts planned for the 2019 New York Auto Show. In addition to revealing the 2020 Sonata...
2020 Hyundai Sonata gets a third-generation platform
Hyundai has already released the first photos of the all-new 2020 Sonata, which we'll get to see debut next month at the...
2020 Hyundai Sonata revealed ahead of its New York debut
After several leaked photos we have our first official look at the 2020 Hyundai Sonata, which is going to debut next month...
The 2020 Hyundai Sonata's Digital Key unlocks the doors with your...
The 2020 Hyundai Sonata is going to debut in April at the 2019 New York Auto Show and now Hyundai has released...
2020 Hyundai Sonata: Here are more leaked photos
Over the weekend the first leaked photos of the 2020 Hyundai Sonata surfaced online and now we have another batch. These two...
2020 Hyundai Sonata leaked
Hyundai has already confirmed that it will unveil the 2020 Hyundai Sonata in April at the New York Auto Show, but now...
2020 Hyundai Sonata will debut at the New York Auto Show
Hyundai has confirmed that the next-generation 2020 Hyundai Sonata will debut at the 2019 New York Auto Show in April. Hyundai hasn't...
2019 Hyundai Sonata Sport loses the turbo
You wouldn't be wrong if you naturally assume that a Sport badge would automatically include a sportier engine, but that's no longer the case...
2018 Hyundai Sonata Eco Review: Sporty and efficient
Midsize sedans aren't getting the attention that they used to years ago, which is why Hyundai gave the 2018 Sonata a big refresh for...

redpajama_set_name: RedPajamaCommonCrawl


The AWS Lofts are located in New York and San Francisco and are an excellent resource for startups and developers for a variety of reasons. One of those reasons is the AWS Architecture Weeks, which are a series of technical sessions that are free for attendees. These single or multi-day events are focused on a specific solution or service, such as Data Analytics, Compute, Architecture, Mobile, Machine Learning, Security, and Databases. Each month, the New York and San Francisco lofts host a variety of these sessions and more. The goal of these event days and weeks is to provide continuous, rich and relevant content to startups and developers.
Interested? October is fast approaching and it's a packed month for the Lofts! Check out the schedule below to learn more, and register to attend an event (or a few!).
Check out Machine Learning Workshop sessions from August at the AWS Loft in San Francisco to get an idea of what's in store!


text:
Breaking It Down: Bias < Fact - Part Three
SpiderManFan2002
"All I or anyone that is speaking out wants is for the Gang Rape gangs to be stopped, thats not racism or islamaphobia."
They are being stopped. Day by day despite the cuts, the police are working arresting people, investigating people- and it's not racism to speak out against it- but it's racist to single out Pakistanis as one ethnic group doing it, it's racist to mock Indians for their poverty, it's racist to say only black guys hit on white girls. And it's Islamophobic to pretend that it's only people who claim to be Muslim commiting these crimes.
"Young white girls are being victimised, please speak out and confront the perpetrators, speak up against those who would silence us& beat us into submission."
Young white girls are being victimised as well as young white boys and BAME girls. It's important we speak up against anyone who acts like the white girls aren't important, or the boys or BAME girls weren't important. They all are important.
"Put yiurself in the victims shoes before attempting to silence us"
Again, I'm not attempting to silence you. Put yourself in a BAME girl's or white boy's shoes, and being constantly dismissed even though the same thing happened to you. Put yourself in a woman or man's shoes who was raped by someone in that 90% statistic and imagine how they must feel if they read that someone sees that statistic as less important. Don't silence those victims.
"imagine that was you or your sister or your daughter."
These are sisters and brothers of the UK <3
"So speak out & let us all unite to cure the disease of hatred. #NoMoreto hatred!"
Yeah! Glad people are picking up the #NoMore thing! Let us all unite and give everyone a voice, let us give the white girls who have been sexually assaulted by anybody a voice! Let us give the white boys who have been sexually assaulted by anybody a voice! Let us give the BAME girls who have been sexually assaulted by anybody a voice! Let us give BAME boys who been assaulted by anybody a voice! Let us give white men and women who have been sexually assaulted by anybody a voice! Let's give BAME men and women who have been sexually assaulted by anybody a voice!
Because if you're apart of the UK, then we're sisters and brothers through nationality, and if you're apart of the world then we're sisters and brothers in humanity!
My Conclusion About The MyTake I Was Reviewing:
You know, if this person said:
"Not all the Pakistani community are like this, so we can't blame them all- but the people who are committing these crimes are predominately Pakistani. There are people of other ethnicities involved in these crimes, for example, white people, African people, Eastern European people."
If this person has made clear:
"Most of the victims are white girls, but there are also BAME girls who are victims there is fewer BAME victims but their voice still matters, and there were white boys who were victims of these grooming gangs, and every victim's voice matters."
If you had stated something like:
"I don't know much about Islam, but I am sure Muslims would not find this kind of abuse acceptable."
If you had also said:
"There are white grooming gangs which exist, and I believe they should be brought to light too- so I will learn more about them"
And if you had not condoned the hateful comments towards Muslims and Pakistanis on your posts I would have been 100% with you.
But the majority of the comments on your posts seem to be more about hating Pakistanis, and hating immigrants, and hating Muslims and Islam, than actually consolidating or helping the victims. I have seen comments consolidating with the victims and about helping the victims on your posts, but I have also seen a lot of comments throwing hate towards Muslims and Pakistanis and that isn't right either.
That is how I know you're motive isn't helping these girls, it's to spark racial hatred.
And using white girls who are victims of rape to peddle your sick fucking agenda, makes me fucking sick! How fucking dare you!
And to everyone else reading this MyTake-
Thank you for reading :)
Part One: Breaking It Down: Bias < fact - part one fact="" -="" part=""></ fact - part one>
Part Two: Breaking It Down: Bias < fact - part two fact="" -="" part=""></ fact - part two>
SincerelyAnon
+1 y
I'm glad someone said something. Other than agreeing with how hateful that post was. I agree there was a sick agenda.
You can't stereotype a whole race of people because you've met some bad eggs. It's ridiculous. Just because someone of whatever race is doing something bad doesn't make them all bad.
Really sad at how hateful some people can be. Whether it's online or in real life.
Also - I hope the victims of all races/genders of people who have been violated in such a way are able to heal & feel safe again. I truly hope they get justice.
Thank you! :) <3
With people like you in the world, we're one step closer :) <3
Thanks! 🙃💚
Discussion: Encouraging women to work in jobs they don't want to do of their own accord.
EmperorOfRussia | 461 opinions shared on Society & Politics topic.
Honest question for you, @SpiderManFan2002:
Do you remember of any of these following quotes?
BECAUSE YOU SHOULD! ALL THESE RAPE APOLOGIST QUOTES CAME FROM PEOPLE YOU ARE FOLLOWING AND TO WHOM YOU ARE VERY AMICABLE!
Your silence is deafening!
YOU IMPOSTER! I'M ASHAMED OF YOUR HYPOCRISY AND EMPTY WORDS.
Enough with your big talk! You are just a BAD ACTRESS.
And how am I in anyway responsible for comments made like that? I didn't say that. I've never seen them before? Who said it and I'll unfollow because I completely disagree with those quotes. Tell me who they are and I will tell them off myself, whichever asshole said this- shouldn't be saying this. Tag them.
EmperorOfRussia
If you dare to delete my comment, I will post a question about this and mention you in the comment section.
@The_White_Queen, I invite you to share your thoughts about the obvious hypocrisy of this Take's authoress.
goaded
Stalking, much?
Impress me by giving links to the quotes, so we can see the context.
Why would I delete your comment I thank you for raising this awareness. Thank you brother.
Please tag the person who said these comments so I can give the bastard a piece of my mind.
Fuck it, I'll find them myself.
@goaded Is there a context to make rape apology?
This is news to me...
I thought you and @24brownish were good friends!
@24brownish I am ashamed of you. How dare you make racist comments as such- these white girls who were raped were far from privileged and non-consensual sex is haram in Islam and is considered Zina. Nobody loves rape, that is why it is called rape.
No white girl or girl for that matter I know wants to be raped. You shouldn't have made those disgusting disrespectful comments to the_white_queen even if you disagree with her. I disagree with her but I would never insult her like this.
I want you to apologise, for offending these people and making these comments. How fucking dare you.
So, they're all from one person? Every one? THAT's context. I've followed a few people on here simply by pressing the wrong button without noticing.
@EmperorOfRussia Well, you thought wrong and I made a mistake following him. Had I had known he was thinking things as such I would have never followed him. I am apologise for causing you offence.
What's your opinion of someone who says:
"I hope this spidermanfan will get raped one day. She is a bitch. Pakistanis are racist because they only rape white girls. I bet she will killed over honour if she wears a bikini or fucks before marriage. You are another reason I am racist against my own race."?
I have a question for you @EmperorOfRussia
What do you think of the quote @goaded posted above?
You made it out like I was following a whole load of "people" saying those things, when in reality I was only happened to be following one- not knowing that he had said these disgusting vile racist things. You assumed we were good friends, and you assumed wrong- and even if we were I would have cut him off the minute I found out he said those racist disgusting things.
And I have unfollowed him, I've called him out, I've block and reported him.
Now somebody has actually wished rape upon me? They have actually said "I hope this spidermanfan gets raped" to me, directly.
So are you now going to condemn her or do you condone what she wished upon me?
And The_White_Queen follows the girl who made this comment, so now are you going post to her what you posted to me?
Are either of you even going to condemn what was said to me?
@S M F,
Do you expect me to be OK with this? Of course I don't think you should suffer sexual violence, or any physical violence either. No matter how it makes me sick your Cultural Marxist way of thinking, you are still a female human - a female human who has regrettable ideas that could harm European cultures if you weren't just a pawn - and people shouldn't wish a barbaric thing like rape upon you.
No woman deserves to be hurt.
And, by the way, I doubt that the person who said that to you - and I don't even know who is - is on my following list. I'm careful to choose the people I interact...
@goaded Funny that you are interested in my opinion about that. What is yours?
It seems I got the person who wrote that mixed up with someone else you are friends with. Sorry.
@EmperorOfRussia You attacked me for following one person- which I did not know said this. I should have been more careful, but then again- I'm 15 years younger than you, so I'm learning as I go.
What I want to know is now The_White_Queen is still following this user (the user is called Shezadi) are you now going to go on to her post- and attack her for following someone who actually wished rape upon me?
Wait a minute, "ALL THESE RAPE APOLOGIST QUOTES CAME FROM PEOPLE..."
People? Who were the others?
@goaded There is another who is a supermoderator and I can't mention. She is very angry at me... I asked a question exposing her hateful words, which she tried to delete but without success because the admins gave a shit about her complaints. Then she asked a question bashing me in the comments, I reported it and it got deleted. LOOOOOL.
Her nickname is a colour plus 2000...
I know who you are talking about. In what way is she hateful?
Some of her words...
But I know that none of those words are a problem to you, Liberal. Hatred is only hatred when it comes from Conservative White men, isn't it?
They sound like jokes, to me, but I'd have to see the context.
Forcing Whites to have children with Blacks is not just rape, but genocide as well.
That person even had the nerve to say something like "if White women got r*ped by Blacks, at least their children would have a culture" (she claims that White people don't have a culture).
@goaded Then what Shez said to SMF could be joke too, according to your logics. Why not?
Let's see (paraphrased).
"People who hate inter-racial relationships will hate inter-racial relationships"
"Those people smell bad"
"I hope this spidermanfan will get raped one day. She is a bitch."
One of these is not like the others.
1st comment of hers: She is talking from her perspective as a black woman. Where did she say she wanted to force white people to have babies with black people? She gave an example of what people who are racist against black people would hate- half black grandchildren. Nowhere did she say that white people should be forced to have babies with black people. And I've witnessed non-white people be racist against black people too.
2nd comment: That was clearly a joke.
I've asked her myself, if she said "if White women got r*ped by Blacks, at least their children would have a culture" I'll see what her reply is.
@goaded No, Liberal. Don't be so disingenuous... She was talking about the smell of rapists. In no way it could be an innocent joke.
It was reported and deleted by the mods before I get to take a screenshot of that. That person even complained why her comments get deleted and mine don't... Well, maybe because I don't make rape apologies...
She said Pakistanis. We're not all rapists.
... In a question specific about Pakistani grooming gangs.
Send me the question.
And her response wasn't about Predominately Pakistani grooming gangs- she didn't say that, she said Pakistanis.
Didn't you just say that "No one is saying that ALL PAKIS ARE RAPISTS"? Is it racist to feel proud of your "White" cultural heritage? ↗
Now you're saying that it's fine to call all Afghans rapists.
wankiam | 4.4K opinions shared on Society & Politics topic.
she called me racist and suggested i condone rape because i pointed out that regulation of local authorities had been tightened up and police had been ordered to investigate every claim since this whole rotheram/rochdale thing came to light and then told me it was the agenda of the left to let this happen even though i pointed out that tory cuts to police numbers and the justice system in general had lead in part to the lack of investigation. also ignored the fact that the outspoken labour mp that got sacked was sacked for an unrelated reason. the problem with these rhetoric shouters on g@g is they never take facts into account but just tend to vilify the person who dares to challenge their view... reading down the page i see you attraccted a couple of them here so if they read this i would point them in the direction of the site guidlines and suggest they learn to use the site as it was intended and not to push their own political agendas.
www.girlsaskguys.com/.../community-guidelines.html
InterServices
Sir would you tell me about the injustice done here?
englisc
"They are being stopped. Day by day despite the cuts, the police are working arresting people, investigating people- and it's not racism to speak out against it- but it's racist to single out Pakistanis as one ethnic group doing it, it's racist to mock Indians for their poverty, it's racist to say only black guys hit on white girls. And it's Islamophobic to pretend that it's only people who claim to be Muslim commiting these crimes."
It is being stopped now. However I think that you're really missing the point, as I suppose those who continue to talk about it because they're not very bright.
The reason people made such a big deal out of the fact that the majority of these rapists were Muslims isn't just because they were Muslims. It's because of the way the authorities did all they could to cover it up for 16 years BECAUSE they were Muslims.
In fact many people who tried to bring this stuff to light had their careers threatened. Some were even threatened that if they spoke out they'd have their kids taken form them. Some were put on trial for "inciting racial hatred" for speaking about these gangs. They wanted to sweep this under the rug and silence anybody who wanted to save those girls because that would harm their own political agenda.
"The reason people made such a big deal out of the fact that the majority of these rapists were Muslims isn't just because they were Muslims. It's because of the way the authorities did all they could to cover it up for 16 years BECAUSE they were Muslims. "
Are you sure they covered it up BECAUSE they claimed to be Muslims, or maybe could have that been an excuse? I'm sure at the time this was happening there were a number people who claimed to be Muslims in prison. So why would one criminal case be different to another?
For years and years they covered up Jimmy Saville. He didn't claim to be a Muslim. For years and years they covered the Jesus Army scandal they didn't claim to be Muslims. They covered up Weinstein and Epstein for such a long time they didn't claim to be Muslims.
In one of these grooming gangs, which I discuss in these series of posts- an MP was involved in grooming young girls. Now when she discovered the grooming gang, she could have easily brought it up in Parliament but chose not to- not because the gang members claimed to be Muslims but to protect her own reputation.
Now this MP was a prominent figure, she ran her own charity claiming to help vulnerable young people.
And in another one of these grooming gangs, some claim a police officer was a prepetrator in some of these gangs. That may or may not be true.
So the question is: How many other prominent figures were involved in these gangs which we may not know of?
That was the reason that many people literally gave when it finally all came out. Journalists, social workers, police officers. They were often threatened by their own employers. So yeah, in each of those there were people involved. That's also the reason that many who tried to speak up were arrested for inciting racial hatred, and some of them were facing up to 10 years in jail each until they were found not guilty. And this was well before dickheads like Tommy Robinson became popular. It was the threat of that which was used to shut people up.
Jimmy Saville was a different case, but equally fucked up. His stuff was covered up because he was rich and well connected.
In each case there were prominent figures involved, and they were covered up for different reasons.
Do you see what they did now?
They used racial hatred as an excuse because it suited their purpose, and at the same time sparked divides within communities. They didn't really give a shit about racial hatred- it just suited them at the time to use that as a front to cover their own backs.
Shezadi | 155 opinions shared on Society & Politics topic.
I am British Pakistani Muslim and I support Tommy Robinson.
And as a Muslim, you should not use swear words.
If you want people to like Pakistanis despite rape gangs then maybe use kindness.
You can't expect the victims or their parents to like Pakistanis.
Stop using the race card.
Imagine, if white gangs were operating in Pakistan?
If you hate white people then leave their country.
" I am British Pakistani Muslim and I support Tommy Robinson."
Well, then you support a racist.
"And as a Muslim, you should not use swear words. "
Oh, so my swear words are worse than racism?
"If you want people to like Pakistanis despite rape gangs then maybe use kindness. "
Oh I am so sorry. Ever since I've been open about my religion, my opinions, facts, my race I experience so much hatred and I've kept calm about it. So I'm so sorry if for once, I retaliate.
"You can't expect the victims or their parents to like Pakistanis."
When the fuck did I say that? Stop twisting what I say. I'm saying it isn't fair people and the media racially stereotype us. Why is it when white grooming gangs abuse people nobody calls them "white grooming gangs?"
"Stop using the race card."
Did you even bother to read the MyTakes I wrote?
"Imagine, if white gangs were operating in Pakistan?"
If I were in Pakistan, and white grooming gangs were operating, but the white community as a whole was being stereotyped- I'd call it out because RACIAL STEREOTYPING IS WRONG.
"If you hate white people then leave their country."
I don't fucking believe this. So calling out racial stereotyping against my own community is now 'hating white people' I don't hate white people, I don't hate anybody. You on the other hand, seem to be a self hater.
@SpiderManFan2002 just ignore or block her. She's not gonna end arguing you until she force you to go to mental hospital with her stupid baseless arguments. She's another @thewhitequeen here who moans here against everyone for her bad experiences in life. Well thewhitequeen is no longer here and banned now looking forward to same thing happening with shezadi.
@InterServices She just changed her account name to use underscores between the words.
I already know and I'm talking about this account. I was told her account has been frozen.
Shezadi
Just look at your own use of language, like how many times did you use the F word? Truth is bitter. You are butthurt because someone mentioned facts and exposed your community. The fact you are triggered shows you have already lost.
One day you will accept that my Pakistani community is toxic. They rape white girls, they justified Qandeel Baloch's honour killing, domestic abuse of Asma Aziz, they shame short clothes, they don't allow girls to have boyfriends or wear what they want and in Pakistan, people can legally get away with honour killings because the male family members can legally forgive the killer.
Victim blaming and shaming happens too. A real Pakistani will speak against Pakistani rape gangs. I am against rape.
I support Tommy Robinson because I am against rapes.
@Shezadi she's not that immature to change her thinking about all Pakistanis whe she gets raped nor will she moan about it here so much about her personal experiences. You don't like Pakistanis or hate something keep it to yourself nobody need to know much. Tell you parents, father, brother and every family member you hate them too for being raised by them because they are rapist, racists and what not.
broncobryan | 171 opinions shared on Society & Politics topic.
Ok here's my opinion on this, yes it's wrong. But how can we not be biased toward a group of people whis religion states that raping young boys and girls is a good thing? In my opinion this is exactly why the UK banned guns. You now have zero way to defend yourselves against anything, you can't even carry a knife for fucks sake.
How can you be biased towards a particular race of people, or a particular religious group of people when it's not everyone in that race who does that crime, and people of other races also commit that crime?
We shouldn't have to defend ourselves because we should have the correct amount of funding in the first place.
I'd say firstly, give more funding to deprived places for things like community centers, libraries, youth clubs- places where young people can take refuge and be safe. Because all these places will have staff which will look out for the younger people, and can inform higher authorities if they suspect a young person is being groomed.
Secondly, I'd say give more funding to police services in the deprived areas of the UK to another way to prevent this because, prevention is better than cure. If more police are patrolling our streets, they can keep an eye out for this kind of thing happening and take procedures in order to prevent it.
Thirdly I'd say it's important we educate young people about this. For example, even though my secondary school was in millions of pounds in debt- they still educated us about grooming and sexual assault and told us what to look out for for ourselves and our friends and who to report it to if we suspect it is happening.
Fourthly, I'd say make the sentencing on this type of thing harsher- and make examples of previous groomers.
Fifthly, I'd say because a number of these groomings happened because of religious extremism we should educate people that no religion condones force or rape or assault etc.
Do you have anything to add? Anything would be helpful.
And you seriously believe guns are good thing? In the USA you have had over 250 mass shootings this year, alone! Think of all the lives that have been lost, the injuries that have taken place, the trauma adults and children have gone through!
broncobryan
250 mass murders? I think your numbers a bit high there. Maybe if you include gun free cities like Detroit, Chicago and New York city.
I said over 250 shootings. I underestimated as a matter of fact. Over 300 mass shootings. An average of 1.24 shootings a day.
en.wikipedia.org/.../List_of_mass_shootings_in_the_United_States_in_2019
WhiteShoulder
Thank you <3. Now I understand.
No problem :) <3
Sevenpointfive | 1.6K opinions shared on Society & Politics topic.
so what do we do? you haven't offered any solutions
Investigate all crimes, without favour.
Sevenpointfive
i looked up pakistani grooming gangs and it led me to sex trafficking. can you explain more about how it works, because you say they get raped and walk off. that's not sex trafficking at all.
we have this thing called neighborhood watch in america. signs in places of high crime that give a number and let people know that others are on the lookout for criminal activity. that only works if they don't want the crime in the area. sometimes a big group that are involved will patrol the streets together. i don't know if it works but it couldn't hurt. you get to know the neighborhood and the people in it really well i'm sure. that's helpful for identifying rapists.
Firstly- what is your search engine? Because I would recommend using DuckDuckGo. On search engines like Google and Bing- it's the paid results which will first come up.
"you say they get raped and walk off"
Where did I say that?
It can happen a numerous amount of ways. Trafficing did happen a lot, I'm not denying trafficking happening. I've seen other cases, where girls were brought back to houses after being groomed to come and then were told to commit sexual acts in order to to stay at the house. Some were invited to parties and assaulted. And in some of these instances, it could have been a one time thing, or a number of these instances could have been trafficking. We can't say for sure- the only people who know these cases properly are the lawyers and solicitors and the people who study these cases word-for-word.
Remember it's the law reports that give us a full picture. Not the news stories, I mean some would say the British media is the least trusted in Europe. News stories can report to us in a way what happens- but law reports are full of nothing but facts.
And please remember it's not just Pakistanis who are apart of these grooming gangs.
As for the neighbourhood watch, we actually do have one of those. I completely forgot about that if I'm honest with you, thank you for reminding me about it.
i guess you don't have the freedom of information act there?
We do have that over here.
then you can buy those reports of those crimes. it's bullshit that they aren't free, but if you want to know what was in the report it might be worth it? you'd find out possibly the houses to stay away from, the people involved and what was done. there's no reason police haven't wiped out this activity other than being paid off.
A lot of reports are kept private and not public because of victim confidentiality- there could be some way to access them for free via the internet, but nothing I know of.
I'm lucky because my college have given us acess to these sites with all the law reports - as law students - for free. But there's a login system and I don't know my login details so hopefully, my teacher should give us them soon.
if you have FOIA then it's a violation. the news gets these stories because they can run down to the courthouse and buy them... lawyers do this too. it's breaking the law if they don't share it, but you might spend a lot of money trying to get it. you could ask a lawyer about that because i don't know much about the process.
I mean I haven't read the full 'Freedom of Information Act' in the UK. I don't know maybe there is some place where the public can access law reports but I don't know about it. I mean I'm assuming there is not, because if there was then why would college pay all that money so that we can access law reports?
We're in two different countries so the legislation for the FOIA may be different.
that's a big something you'll need to look into before you become a lawyer. good luck
AkshiJanjua | 324 opinions shared on Society & Politics topic.
i think you should just let it be and stop explaining people and just let it be because some people will never understand and it stems from racist ideologies of people which have been perpetuating in their minds from centuries ago and particularly in UK its just how history has been for them and they may feel those Asian immigrants which their ancestors hated were probably evil because of these events that are taking place there. Its a matter of projecting a whole race, religion, culture wrong because some people among are wrong.
Be proud of your heritage and keep doing what you love time will tell who your real friends are who really care and respect you.
88-mph
Sounds like your biased against the victims
Home > Society & Politics > Breaking It Down: Bias < Fact - Part Three
Would a person be more mad they must leave on a certain date or eviction?
They're mad because they're in fact leaving on that date
They're mad because they're staying and angry they will face eviction


The fans have been electric since United's move to Audi Field. The team clinched the playoffs with a 3-1 win against NYCFC.
In recent months, D.C. United has made waves in the soccer community. After years of playing in the run-down RFK Stadium, D.C. United has finally moved into their new home at Audi Field. The 20,000 seat, $500 million stadium, coupled with the surprise signing of English superstar striker Wayne Rooney, has helped propel United into an astonishing playoff spot in the final months of the season.
Among fans at WJ, many believe the numerous changes at D.C. have positively affected the team. Junior Camille Lukash has season tickets at Audi Field and believes.
"Fan involvement has definitely had a positive increase since the inaugural game at Audi Field. The attendance has risen and fans are a lot more spirited and hype during games," Lukash said.
Additionally, Lukash prefers Audi Field to the old, deteriorating RFK Stadium because it is more modern and open, so there aren't really any bad seats.
Before signing Rooney, United sat in last place in the league with 11 points. Now, three months later, they have confirmed a playoff spot and could finish anywhere from 6th to 4th place depending on other results in the league. Rooney has been widely credited with United's impressive performance, but it is important to acknowledge the other players such as midfielders Luciano Acosta, Russell Canouse and Paul Arriola, who have all flourished alongside their new captain. Rooney's advanced knowledge of the game has allowed him to play a strong leadership role, and has changed the team's style on and off the field.
Freshman Darius Kamrad believes the signing of Rooney has had the biggest impact on the team and that he would definitely be considered United's player of the season.
"He is a great leader, player and communicator. He works very well with his teammates and D.C. United. [Improving from last to sixth place since the signing of Rooney] is proof that Rooney has impacted the team," Kamrad said.
The Englishman plays as more of a reserved striker or "false nine", meaning he usually has his back to the goal and is able to be involved in a lot of the build up play. This allows midfielders such as Arriola and Acosta to make sharp inward runs towards the goal instead of relying solely on Rooney's finishing ability. The trust he has shown in his new teammates has also made Rooney an instant fan favorite. From the start, Rooney rejected the number 10 shirt (which he famously donned at Everton and Manchester United) when he signed for the club, saying it belonged to Acosta and instead opted for number nine. He additionally rejected first class travel from the club so he could bond with his new teammates, hoping that he could be seen as one of them rather than a world class player on another level. This commitment to becoming part of the team and adapting so well to life in DC is what has fans on the edge of their seats, eager to see what's next for United.
Junior Arturo Alipio agrees with Kamrad about Rooney's ability and positive influence on the team, citing "that crazy play against Orlando City" in which Rooney chased down an attacker for 50 yards with infinite space and an open goal ahead of him. He risked everything by diving in with a slide tackle, won the ball, played a 40 yard lob to Acosta who managed to jump over the defender to head it in to win the game in the 96th minute.
"I appreciate the work rate Rooney brought to DC," Alipio added.
United play their final regular season game against Chicago Fire on Sunday, October 28.


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Enjoy Great Music at Tuscaloosa's Live at The Plaza Concert Series
Dre Day Published: June 2, 2021
Alenate
Enjoy Great Music at Tuscaloosa's Live at The Plaza Concert Series
It's the summertime and if you're looking to get out of the house, this is the perfect event for you and the family. The city of Tuscaloosa's Live At The Plaza concert series is back!
Friday, June 4, 2021, the City of Tuscaloosa will kick off its free live music concert series in Government Plaza. These are family-friendly events for all to enjoy. After the kind of year we had in 2020, it's no doubt that things like this are missed in Tuscaloosa.
The City of Tuscaloosa will have a total of 9 summer concerts featuring a lot of great acts. The schedule was released in an official press release.
AL – The City of Tuscaloosa's Live at the Plaza summer concert series returns this Friday, June 4, from 6 to 9 p.m. in Government Plaza. Kemistry and Sonik Jones will be performing.
The Live at the Plaza summer concert series presented by Coca-Cola and Alabama Credit Union features free live music every Friday in June and July. The event is family-friendly, and free parking is available in the adjacent Intermodal Facility Parking Deck.
The lineup for this summer's concerts will be:
June 4: Kemistry, Sonik Jones
June 11: Jus Gruv Entertainment, The B-Sides
June 18: Algorhythm, Casey Thrasher
June 25: Keith "Cashmere" Williams, Justin Oliver
July 2: The Powell Brothers, Measures of Loud
July 9: The Locals, The M-80s
July 16: Stonegrey, One Wild Sunday
July 23: Ula - A Tribute to Jimmy Buffett, Amanda Shaw
July 30: The Locked Band, The Jous Band
For more information about Live at The Plaza, click here.
TACKLE THESE: Check Out the Best Uniforms In Each NFL Team's History
Filed Under: Featured, Government Plaza, Live at the Plaza
Categories: Articles, Local News, News, Tuscaloosa Events


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Highlight from the series of events the "Counseling Week of Studying Abroad 2022"
The series of events "Counseling Week of Studying Abroad 2022 with ISA", which lasted 4 weeks from February 12th to March 5th, 2022, officially ended with the participation of a large number of students. The majority of participants are students in grades 11 and 12 in Ho Chi Minh City who intend to study abroad after graduating from high school in Vietnam.
In contrast to the "bleak" situation two years ago, the enrollment period of colleges and universities around the world is busier this year, as many of the epidemic's restrictions have been removed. Furthermore, many opportunities have been opened up in 2022. One of these is that some governments and universities have increased their budgets for financial aid and scholarship packages while reducing pressure on procedures and academic requirements. Moreover, thanks to the strategic cooperation relationship between Vietnam and other countries, Vietnamese students have more opportunities than others.
With these benefits, many young people quickly seized this chance to study abroad in 2022 to carry out their long-delayed dream. Participating in the event "Counseling Week of Studying Abroad 2022 with ISA", some students expressed their regret at delaying their plan of studying abroad because of the effects of the pandemic. Most of them also wished to have a clear plan so that they can stay overseas to study, work and even settle down.
At the events, ISA's consultants assisted the students in preparing for the upcoming admissions in 2022. Besides, with a global network of partners all over the world, ISA has also provided opportunities for potential students to apply for scholarships to some of the world's top universities, including The University of Adelaide (one of Australia's Go8 representatives), Niagara College (Canada), Illinois State University (USA), etc.
The event was also a chance for ISA to meet parents and students as well as understand their concerns in planning for studying abroad in the next years. It is expected that in the future, ISA will continue to organize many other programs and events to bring the latest updates on studying, working, and settling in other countries such as the US, Canada, and Australia,…
Parents and students who want to learn more about studying abroad, scholarships, and visas, please contact 0938 938 748 for a free consultation.


Robert John "Robin" Finck (born November 7, 1971) is an American guitarist. He is one of only a few artists who has played in two different bands listed on VH1's "100 Greatest Artists of Hard Rock": Nine Inch Nails (ranked no. 43]) and Guns N' Roses (ranked no. 9).
Finck grew up in Marietta, Georgia and played with several unsigned bands based primarily in the Atlanta area, including acts such as Prowess, Bat Your Lashes, Sik Dik (bandmates of which included Steve "Holiday" Childress and Michael Allen, currently of The Goodies and Greg Thum), and Impotent Sea Snakes.
He joined Nine Inch Nails as part of their touring band in 1994–95 for their Self-Destruct and Further Down the Spiral tours following the departure of previous guitarist Richard Patrick. Finck appeared with Nine Inch Nails at Woodstock '94, and saw his first official release with them, the Closure video, released in 1997. Once the tour was completed, he took a job playing for the circus troupe Cirque du Soleil in the original tour of Quidam. In 1997 Finck signed a two-year contract with Guns N' Roses as the replacement for Slash (who had quit the band at the end of the previous year) and began work on what was ultimately to become the band's long delayed album Chinese Democracy. When the two years were up the album had not been completed, although the song Oh My God was released on the End of Days soundtrack. Whether or not Finck played on the song is in dispute; GNR vocalist Axl Rose says he did while Finck himself says he did not.
Finck rejoined Nine Inch Nails as the band embarked on their Fragility v1.0 and Fragility v2.0 tours in support of the 1999 album The Fragile. These tours were recorded and released as the live album/DVD, And All That Could Have Been, which marked Finck's second release with them. He also played on a studio-recorded version of The Day The World Went Away on the companion album Still, which was included with the deluxe version of And All That Could Have Been and later made available as a standalone CD through nin.com. Shortly after the tour ended in 2000 he returned to Guns N' Roses, playing four shows with them in late 2001 followed by a brief tour of Europe and Asia in mid-2002 and a live appearance at the MTV Video Music Awards. A planned US tour was meant to continue through the remainder of 2002 but was called off by the promoter after two cancelled shows led to rioting by fans. Around this time Finck and then-bandmate Buckethead contributed to the soundtrack of the John Carpenter movie Ghosts of Mars, playing alongside members of thrash metal group Anthrax and guitar virtuoso Steve Vai. After his touring obligations with GNR were fulfilled he joined the LedZAriel production (a performance by acrobats set to the music of the band Led Zeppelin).
Finck was invited to rejoin NIN to tour in support of their 2005 album With Teeth but chose not to and the position was filled by Aaron North. In the summer of 2006, he toured Europe with Guns N' Roses playing in front of over 700,000 people. They finished up by touring North America for the remainder of the year, and went to Mexico, Australia and Japan in 2007.
On April 4, 2008, it was announced via nin.com that Finck would be rejoining Nine Inch Nails as a member of the live band. He subsequently contributed to the Nine Inch Nails album The Slip, earning a performance credit alongside then-band members Josh Freese and Alessandro Cortini.
In November 2008 Guns N' Roses' long awaited album Chinese Democracy was released. Finck has a co-writing credit on the song "Better", which was later released as a single. He also played guitar on every track, performed 7 of the guitar solos and has additional credits for keyboards, arrangements and pre-production. He chose to remain with NIN after the album's release and, as of March 21, 2009, has been replaced as the lead guitarist of Guns N' Roses by DJ Ashba. The official Guns N' Roses website states that "Robin continues to be part of GN'R, by virtue of Guns' history and his involvement in Chinese Democracy".
Following the departure of NIN live band members Alessandro Cortini and Josh Freese at the end of 2008 Finck and bassist Justin Meldal-Johnson re-grouped with band leader Trent Reznor and new drummer Ilan Rubin and the 4-piece band completed an 8-date tour of Australia and New Zealand between February and March 2009. Finck continued to perform with the band throughout the 2009 Nine Inch Nails/Janes Addiction (NIN|JA) tour, with shows throughout North America, and the subsequent Wave Goodbye tour through Europe and Asia. The band then returned to the US to play a series of shows culminating in the final live appearance of Nine Inch Nails.
In October 2009 Finck and former Guns N' Roses bandmates Axl Rose, Tommy Stinson and Buckethead were named in a lawsuit by electronic musician Ulrich Schnauss's record labels Independiente and Domino alleging that they had committed copyright infringement by using portions of his compositions in the song "Riad 'n the Bedouins". Finck appears on the 2010 album The Lady Killer by artist Cee-Lo Green, performing guitar on the track "The Lady Killer Theme (Outro)".
On March 12, 2012, Finck appeared on-stage with Guns N' Roses, performing "Better", a song he co-wrote with Axl Rose.
Well, when I was first playing with Robin, I was new in the band and I think it's the kind of thing, it's like, I got the feeling it was almost like mom just got home from the hospital with the newborn. It's like, "Here's your new brother, play nice. The kid is like, "Who's this?" Just the skepticism and the doubt and the whole, "I'm the son, not you," kind of, just all the things that come along with that situation, where suddenly someone else is thrown into your world.
It's like, "Wait a minute. Don't I have a say in this? This is my family, too," kind of vibe, just the little elements of that. So it was really rocky at first and for a long time. So, I think that me and Robin, probably after he left the band, we became better friends. When he was playing with Nine Inch Nails and I would go to see him with that and we'd hang out after the show, and I think we became closer after we stopped playing together because of the way we were thrown together.
Because, at the time, they were auditioning people for months and they were so burnt out. The tour was a few weeks away. They thought that, "All right, we're not getting the third guitarist like Axl wants," which was always his vision. He always wanted a three-guitarist band. He loves Boston. He loves a lot of the old classic stuff. He's just a guitar lover.
Apparently, Robin is back in NIN.
The first in about two years.... Anyone knows what he is talking about?


Standard Chartered Bank is strengthening its mining and metals offering in response to the recent high growth within this segment of the commodities sector. The bank has established a team of client relationship specialists and also expanded its commodity corporates, corporate finance and commodity research teams.
New York-based Karr McCurdy has been appointed as global head of commodity corporates – mining and metals to steer client relationship teams across Europe, Africa, Asia and Americas.
Karr has almost 30 years of international experience in the mining and metals sectors. Working first as an engineer and then as an advisor and financier, Karr has assisted companies and governments on developing and managing new and expanding projects globally.
Douglas Strong, formerly Standard Chartered's head of mining and metals, project and export finance has been appointed as managing director, regional head of commodity corporates, mining and metals for Europe and Africa, based in London.
Strong has over 19 years of international banking experience, 15 of which have been involved with mining and metals globally. He has been involved with project and structured financing, acquisition financing and bid advisory across precious and base metals.
Dan Miller has been appointed regional head of commodity corporates, Asia Pacific, based in Singapore. His 15 years experience in the mining and metals sector spans equity research and corporate banking.
Miller has worked closely with companies in the mining and metals sectors to assist them in major fund raising activities, acquisition finance, working capital needs, risk management strategies, and transactional banking needs.
Additional new hires have also been made in the Middle East, South Africa and New York to complete the global coverage model.
Complementing the specialised client industry and advisory team, Corporate Finance has appointed David Trafford as the Global Head of Mining and Metals, Project and Export Finance (PEF) based in London.
David Trafford has more than 20 years experience in the mining and metals sector, advising companies and governments on strategy, financing options, acquisitions and disposals in public and private transactions. Trafford's experience spans base metals, precious metals, coal mining and bulk commodities working in the mining and metals industry and banking. He has provided marketing for and executed mandates in across the globe, particularly in Africa, the Americas, Europe and Asia.
Additional new PEF hires have also been made in China, Dubai and South Africa to complete the global coverage model.


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About ATUS-X
About ATUS
Donate to IPUMS
Browse or Select Data
Analyze Data Online
Time use variables
Download or Revise My Data
Sample Descriptions
Link to CPS
Discover IPUMS Variables
Citing IPUMS ATUS
IPUMS Bibliography
This information has been created by the Centre for Time Use Research at the University of Oxford
Study title:
American Time Use Survey (ATUS) 2003
Bureau of Labor Statistics, USA Department of Labor
When conducted:
Whole year 2003
Sampling method and study design:
This study aims to show what people across the USA are doing on any given day. This year is the first year of continuous time use data collection in the USA. The sample in this initial year was particularly large, though have been reduced for future years. The continuous data can be pooled to create a larger sample for analysis. Procedures for pooling are described on the ATUS website.
The ATUS is a national sample of adults aged 15 or older in private households (not nursing homes or institutions), and is drawn from a stratified three-stage random sub-sample of households that have completed the 8th and final wave of the Current Population Survey (CPS). The diary wave takes place around two months after the final CPS interview.
The diaries were collected from one person per household about the day before the interview (yesterday) using computer assisted telephone interviewing (CATI). Participants were assinged to complete a diary for a particular day of the week. If people were sampled for a Monday and could not complete the diary on the first Monday of attempted contact, then the interviewers tried to call again for information about another Monday. The interviewers tried to contact people for 8 weeks. Half of the diaries were collected on weekend days, and half on week days. Respondents were contacted by mail with information about the study before being phoned for the diary interview. Around 5% of CPS households did not have a phone. These households are included. People in households with no phone were sent phone cards to enable them to call the survey without charge. Interviews were conducted in English and Spanish.
The study collect one 24 hour diary from participants. Interviewers recorded activities in respondent's own words. The diaries started at 4AM, and respondents told the interviewer the time that they started and stopped each activity. The diary collected main activity only (no secondary activities), who else was present, and location/mode of transport. If people who looked after a child aged <13 on the diary day, respondents were asked to indicate during which activities a child aged <13 was in their care (paid work, sleep and grooming excluded).
The software and the interviewers used some diary information to fill in some gaps - for instance to note that an activity was travel is the diarist reported a mode of transport but no activity.
20,720 respondents
Response rate:
Weighting procedures:
The data include weights which adjust for oversampling of some groups, adjust for non-response, and balance the days of the week
Sources of information:
Available documentation:


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The city of Rockford, located in Winnebago County, Northern Illinois, is home to just over 150,000 inhabitants. Although currently the third-largest city in the state, it was once the second-largest in the latter part of the 20th century. Also known as the "Forest City" because of its famous oak trees, Rockford was initially called "Midway" because of its mid location between Chicago and Galena. It was quickly named Rockford after it offered an excellent crossing point for wagons over the Rock River. With yards of woodlands, parks, and recreational areas, Rockford has copious outdoor and recreational activities, such as hiking, trekking, fishing, biking, and sight¬seeing.
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