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Apple Caught Using Photoshop to Fake More Pics in Lawsuits
Jason Mick
August 19, 2011 3:05 PM
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Apple has creatively doctored another photo to try to make Samsung look guilty of design patent infringement.
(Source: Apple via IDG)
Apple CEO Steve Jobs says his company is a master of stealing ideas. Apparently it's also becoming masters of using Photoshop to provide potentially perjurous doctored exhibits in court.
(Source: Sydney Morning Herald)
Using doctored images, Apple succeeded in convincing a German judge to ban sales of the Galaxy Tab 10.1 in Germany.
(Source: HiTech)
You're viewing it wrong!
Apple, Inc.'s (
) chief executive and co-founder, Steven P. Jobs has bragged about his mastery of stealing ideas from others,
[video], "Picasso had a saying - 'Good artists copy, great artists steal.' And we have always been
about stealing great ideas."
I. Apple Alters Reality With Photoshop
But stealing other competitors' ideas isn't the only thing Apple is good at. Of late it's shown itself to be a burgeoning master of Photoshop. It was caught in Germany
doctoring images
(playing with perspectives and stretching pictures) to make its competitor's product look like its own.
Now Apple has been caught yet again engaging in egregious editing images -- quite literally "stretching the truth" -- in the Netherlands.
The Netherlands court filing came as part of Apple's legal crusade [
] against Google Inc. (
) and its hardware partners.
Unable to compete
against Google's Android operating system on the free market, Apple has come up with the creative solution of trying to use lawsuits to remove Google from the market altogether.
In this particular case, Apple's victim was Samsung Electronics Comp, Ltd. (
). Apple accused Samsung of "slavishly" copying the iPhone 4. It sued trying to ban Samsung's widely successful Galaxy S smart phone family from the market in the Netherlands.
But it appears, in reality, that it was Apple who was "slavishly"
copying and pasting
the Galaxy S into Photoshop. The Galaxy S is a bit larger than the iPhone 4 (122.4 by 64.2 mm) and looked decidedly different, so Apple had its work cut out for it in order convince a judge that the phone was "copying" the iPhone's patented look -- a key claim in its case.
So it shrunk the Galaxy S down to iPhone size -- 115.5 by 62.1 mm. It also cherry-picked an image of the iPhone 3G, a defunct model it has not produced since 2009, as that model had more rounded edges -- like the Galaxy S -- and unlike Apple's current production iPhone (the iPhone 4).
A careful read would reveal flaws in the visual exhibit -- Apple did at least admit in the text, that the Galaxy S has "some non-identical elements, such as the slightly larger dimensions."
But consider that the claims of design patent infringement are critical to the case, and that the picture came on page 77, it's possible by that point that a weary justice might not be reading the small print very carefully.
II. Images Played Key Role in Sales Ban
The recent incident, like the previous photoshopping, was
by Dutch IDG publication
. IDG quotes Mark Krul, a lawyer at the Dutch firm WiseMen and a specialist in IT and intellectual property law, who says the visual evidence was critical in a German court's decision to
ban Samsung's tablet sales
in the European Union via an
ex parte
Mr. Krul was astonished that doctored images appeared in a separate filing,
, "It surprises me that for the second time incorrect presentations of a Samsung product emerge in photographic evidence filed in litigation. This is not appropriate and undermines Apple's credibility both inside and outside the court room. Apple has certainly some explaining to do, if only to clear itself from the appearance of improper behavior."
The EU court has since changed its ruling to only banning sales in Germany. And Samsung is actively fighting to reverse that ban. A Samsung lawyer in the EU, Bas Berghuis of Simmons and Simmons, accused Apple of "manipulating visual evidence, making Samsung's devices appear more similar to Apple's."
A legal expert who spoke to IDC anonymously said that in patent law cases often visual exhibits are the main thing judges look at, as they're considered the point that the plaintiff wants to emphasize -- more so than the text.
While the discovery of its "creative" alterations may be bad news for Apple's legal chances, on the plus side its legal team may be able to find jobs for themselves as graphics designers, should they lose the case. After all, they're building a pretty impressive portfolio of work, already.
"So, I think the same thing of the music industry. They can't say that they're losing money, you know what I'm saying. They just probably don't have the same surplus that they had." -- Wu-Tang Clan founder RZA
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Removing Noise From Pyrosequenced Amplicons
- Christopher Quince1Email author,
- Anders Lanzen2,
- Russell J Davenport3 and
- Peter J Turnbaugh4
© Quince et al; licensee BioMed Central Ltd. 2011
Received: 21 June 2010
Accepted: 28 January 2011
Published: 28 January 2011
In many environmental genomics applications a homologous region of DNA from a diverse sample is first amplified by PCR and then sequenced. The next generation sequencing technology, 454 pyrosequencing, has allowed much larger read numbers from PCR amplicons than ever before. This has revolutionised the study of microbial diversity as it is now possible to sequence a substantial fraction of the 16S rRNA genes in a community. However, there is a growing realisation that because of the large read numbers and the lack of consensus sequences it is vital to distinguish noise from true sequence diversity in this data. Otherwise this leads to inflated estimates of the number of types or operational taxonomic units (OTUs) present. Three sources of error are important: sequencing error, PCR single base substitutions and PCR chimeras. We present AmpliconNoise, a development of the PyroNoise algorithm that is capable of separately removing 454 sequencing errors and PCR single base errors. We also introduce a novel chimera removal program, Perseus, that exploits the sequence abundances associated with pyrosequencing data. We use data sets where samples of known diversity have been amplified and sequenced to quantify the effect of each of the sources of error on OTU inflation and to validate these algorithms.
AmpliconNoise outperforms alternative algorithms substantially reducing per base error rates for both the GS FLX and latest Titanium protocol. All three sources of error lead to inflation of diversity estimates. In particular, chimera formation has a hitherto unrealised importance which varies according to amplification protocol. We show that AmpliconNoise allows accurate estimates of OTU number. Just as importantly AmpliconNoise generates the right OTUs even at low sequence differences. We demonstrate that Perseus has very high sensitivity, able to find 99% of chimeras, which is critical when these are present at high frequencies.
AmpliconNoise followed by Perseus is a very effective pipeline for the removal of noise. In addition the principles behind the algorithms, the inference of true sequences using Expectation-Maximization (EM), and the treatment of chimera detection as a classification or 'supervised learning' problem, will be equally applicable to new sequencing technologies as they appear.
Next generation sequencing has revolutionised many areas of biology by providing a cheaper and faster alternative to Sanger sequencing. One technology that is finding many applications, for example in de novo genome sequencing, or diversity studies of regions of DNA that have been amplified by PCR, is 454 Pyrosequencing [1]. It is this latter application of 454 to the sequencing of PCR products or amplicons that we will focus on here. 454 Pyrosequencing generates large numbers of reads, 400,000 in the original GS FLX implementation increasing to 800,000 with Titanium reagents, which are long compared to other pyrosequencing platforms, 250 bp for GS FLX increasing to around 500 bp for Titanium. This makes it ideal for high resolution studies of the sequences and their relative frequencies in relatively long PCR products. The method is to simply sequence the diverse amplicon sample without cloning individual sequences. This has many applications for instance in viral population dynamics [2], or characterising microbial communities through amplification of 16S rRNA genes [3].
Per base error rates from 454 pyrosequencing are comparable to those from Sanger sequencing [4] but without cloning resequencing is impossible. In addition, the large read numbers obtainable mean that the absolute number of noisy reads is substantial. Consequently, it is critical to distinguish true diversity in the sample from noise introduced by the experimental procedure. This is particularly true if we want to calculate the absolute number of different sequences, or clusters of sequences, present. This is effectively the problem in microbial diversity estimation, where sequences are clustered into Operational Taxonomic Units (OTUs) that proxy for traditional taxa and we are interested in estimating the number of such OTUs in a community. It has already been noted that noise in 454 amplicon sequencing leads to inflated estimates of OTU number [5, 6]. This is important because surprisingly large OTU numbers together with a bias towards rare taxa were observed in the first studies of pyrosequenced 16S rRNA genes [3, 7]. This preponderance of rare taxa has been termed the 'rare biosphere'. The spurious OTUs generated by noise generally have low frequencies consequently noise may explain both the high OTU numbers and the bias towards low abundances reported. Development of effective noise removal algorithms is therefore a matter of urgency in the exploration of microbial diversity.
PyroNoise is a relatively sophisticated algorithm that reconstructs the true sequences and frequencies in the sample prior to OTU construction using a mixture model [5]. It is based on clustering flowgrams rather than sequences which allows 454 errors to be modelled naturally. Using this approach it is possible to account for two facts: firstly that sequences with errors are likely to be rare and secondly that they should be similiar to a true abundant sequence. The mixture model approach allows this to be done in a very natural way by fitting noise distributions around each proposed true sequence. The drawback is that an iterative and hence computationally costly algorithm must be used.
Two sources of error need to be considered in pyrosequenced amplicons. Those from the pyrosequencing itself and those introduced by the PCR amplification. The original implementation of PyroNoise simultaneously removed both sources of errors. Consequently it was necessary to align flowgrams. Here we present a new approach, AmpliconNoise, which couples a fast flowgram clustering step without alignment, still called PyroNoise, to a sequence based clustering, SeqNoise, which does perform alignments. The latter explicitly accounts for the differential rates of nucleotide errors in the PCR process, and uses sequence frequencies to inform the clustering process. The result is a more sensitive program than the original PyroNoise achieved at lower computational cost because the fast alignment free flowgram clustering reduces the data set size for the slower sequence clustering. AmpliconNoise has already been used to determine gut microbial diversities [8] and for viral population genetics [2].
Recently another flowgram based denoising algorithm, DeNoiser, has been developed [9]. This was motivated by the original PyroNoise and uses the same flowgram alignments but incorporates several modifications to increase speed. It begins by finding unique sequences, orders them by frequency, and then starting with the most abundant maps the other reads onto these 'centroids' if their distance to the centroid is smaller than some threshold. The distance used is the same flowgram based measure as in the original PyroNoise. It is therefore a greedy agglomerative clustering rather than iterative. This reduces the computational costs of the algorithm but misassignment of reads when the true sequences are similar, may result in a loss of ability to accurately reconstruct OTUs. An even faster approach is to perform the same centroid based clustering using sequence rather than flowgram based distances, this is referred to as single-linkage preclustering (SLP) by Huse et al. [10], and a similar strategy is adopted in the PyroTagger program [11]. In this paper we will describe AmpliconNoise fully for the first time and compare to the original PyroNoise algorithm, DeNoiser, and SLP in terms of ability to remove noise and allow accurate OTU construction. We will use 454 pyrosequencing data from known sequences for these evaluations including both standard GS FLX and newer Titanium data.
SeqNoise accounts for PCR single base errors but the PCR process can also introduce sequences that are composed of two or more true sequences, so called 'chimeras'. These generate sequences that are quite different from either parent and so can not be removed by clustering. Chimeras are generated when incomplete extension occurs during the PCR process and the resulting fragment effectively acts as the primer in the next round of PCR. Existing algorithms for removing chimeras were developed for full length clone sequences and lack the sensitivity for short pyrosequencing reads [12, 13]. The program ChimeraSlayer is to our knowledge the only current chimera checker capable of handling 454 reads effectively. ChimeraSlayer requires a reference data set of sequences that are known to be non-chimeric [14]. We introduce a new algorithm, Perseus, based on two novel principles for chimera removal: firstly because the parents of any chimera must experience at least one more round of PCR than the chimera then we can search amongst all those sequences of equal or greater abundance than the chimera for possible parents; secondly that chimera removal should, with suitable training data sets, be treated as a problem in classification or 'supervised learning'. Adopting these principles Perseus has the sensitivity to remove chimeras from 250 bp GS FLX reads with the advantage of not requiring a set of good reference sequences.
We will now demonstrate that AmpliconNoise followed by Perseus is capable of removing the vast majority of erroneous reads from 454 pyrosequencing data, reducing overall error rates, and thereby allowing accurate OTU construction and microbial diversity estimation.
Test Data Sets
To test the noise removal algorithms we used eight previously published test data sets and one hitherto unpublished. These were all generated by preparing mixtures of known DNA sequences in known concentrations and amplifying and pyrosequencing. For the published data sets the standard GS FLX protocol was used but specifically for this study we generated a further data set with the most recent Titanium reagents. Three different 16S rRNA regions were amplified in all cases with a standard Taq polymerase, the V2 region [8], the V5 region [5], and for the Titanium data V4-V5. The mixtures consisted either of 16S rRNA clones in the case of the V5 and V4-V5 data sets or DNA extracted from 67 separate isolated organisms for V2. Two V5 data sets were prepared: one 'Divergent' data set comprising 23 clones that differed at a least 7% of nucleotide positions mixed in equal proportions, facilitating the unambiguous mapping of each read to a known clone, and one 'Artificial Community' where some clones differed by just a single nucleotide and concentrations varied by two orders of magnitude mimicking a natural community. The V2 'Mock Communities' were similarly split between three replicates where the extracted DNA was mixed in equal proportions (Even1, Even2, Even3) and three where it was mixed unevenly (Uneven1, Uneven2, Uneven3). Full details of these test data sets are available from the original publications [5, 8].
The Titanium data was generated by pyrosequencing a mixture of 91 full length 16S rRNA clones obtained from an Arctic soil sample. These clones were independently Sanger sequenced although only 89 sequences could actually be recovered. Consequently the results presented here will be a lower bound on accuracy with a few sequences falsely categorised as errors that should be in the sample. Since this will apply equally to all the tested algorithms our ability to compare between them is not affected. The mixture contained each clone in equal abundance. This DNA mixture was then pyrosequenced following amplification with 16S rRNA primers that also had a tag (AGTGCGTA) and the standard Titanium A and B adaptors attached. The primers used were both degenerate, F515 (GTGNCAGCMGCCGCGGTAA) and R926 (CCGYCAATTYMTTTRAGTTT). Sequencing was forward from F515 so as to capture the V4 and most of the V5 region with a 400-500 bp Titanium read.
The eight GS FLX and one Titanium test data sets used in this study
Name | Type | Abundances | 16S region | Read no. | Filtered no. | DeNoiser no. |
Divergent | 23 clones | Even | V5 | 57,902 | 35,190 | 42,052 |
Artificial | 90 clones | Uneven | V5 | 46,249 | 31,867 | 37,903 |
Mock Communities | ||||||
Even1 | 67 genomic | Even | V2 | 63,780 | 53,771 | 55,398 |
Even2 | DNA isolates | Even | V2 | 53,763 | 45,178 | 46,294 |
Even3 | DNA isolates | Even | V2 | 67,182 | 54,153 | 55,797 |
Uneven1 | DNA isolates | Uneven | V2 | 54,099 | 44,926 | 46,837 |
Uneven2 | DNA isolates | Uneven | V2 | 51,439 | 44,176 | 44,880 |
Uneven3 | DNA isolates | Uneven | V2 | 60,976 | 50,931 | 53,225 |
Titanium | 91 clones | Even | V4 - V5 | 62,873 | 25,438 | 21,477 |
Origins of Pyrosequencing Noise
Filtering Noisy Reads
It has previously been shown that some features correlate with noise in reads from 454 data [4, 6]. Consequently filtering for those reads can reduce the overall level of noise in the data set. However, substantial noise remains after this process [5]. The purpose of this study is not to evaluate different filtering methods for reads, but rather to address the problem of how to account for this remaining noise. Consequently we adopted a rather strict filtering procedure: first we checked for an exact match to the primer and tag if present, we then used the observation that signal intensities between 0.5-0.7 are associated with noisy reads [4]. We therefore truncated all reads at the first such signal, or any sequence of the four nucleotide flows that failed to give a signal ≥ 0.5, any read where this occurred before flow 360 of the 400 flows in a GS FLX run we removed. For the Titanium reads which have 800 flows we used the same procedure keeping only reads where the first noisy flow occurred on or after 360. In addition, the level of noise in reads increases towards the ends of reads [15], to account for this we removed the last 10% of flows truncating all GS FLX reads at flow 360 and Titanium at 720. The numbers of reads after filtering for each data set are given in Table 1.
Removing Pyrosequencing Noise
to generate a total distance which is simply the sum of the distances for each signal. We used the V2 distributions, Figure 1, to calculate flowgram distances for all the GS FLX data sets but calculated new distributions for the Titanium data.
To infer the true sequences and their frequencies we maximise this likelihood using an expectation-maximization (EM) algorithm. EM algorithms apply very naturally to complex clustering problems. Intuitively they exploit the fact that if the properties of the cluster centres are known, then the probability that a given data point was generated by a given cluster centre is easy to calculate, similarly if those probabilities are known then maximum likelihood solutions for the parameters of the cluster centres can be calculated. What is hard to do are these two steps simultaneously. In an EM algorithm we avoid that by iterating the two steps separately until the parameters converge at a local maximum of the likelihood [16]. The algorithm used here differed in two ways from that proposed previously [5]. Firstly, we did not align flowgrams to our denoised sequences before calculating the distances. That was necessary to allow for PCR errors that cause changes resulting in insertions and deletions at the flowgram level. Flowgram gaps do occur very rarely as a result of pyrosequencing noise, occasionally no signal ≥ 0.5 is observed in a frame of four nucleotides, but as described above we truncated our flowgrams when this was observed. Consequently by not performing alignments we ensured that only pyrosequencing noise would be filtered at this step. Secondly, we did not construct the maximum likelihood sequences each flow at a time, instead we only allowed sequences that were observed in the data. This allows the final denoised reads to be mapped to the originals.
- 1.M step: Given the set each sequence to the sequence corresponding to the perfect flowgram Ū j that maximises Equation 3, restricted to the set of P unique perfect flowgrams obtained by rounding the observed flowgrams to integers. This corresponds to finding the perfect flowgram with the smallest total distance to all the reads weighted by the conditional probabilities that each flowgram was generated by that denoised sequence:(4)
- 2.
Calculate new distances .
- 3.E step: Calculate new as:(5)
- 4.
Repeat until convergence
Expectation-maximization algorithms because they only find local optimum are sensitive to initial conditions. To initialise the EM algorithm we performed a complete linkage hierarchical clustering based on flowgram distances and formed clusters at a given cut-off, c p . This also defines the number of denoised sequences L, although the number with non-zero weight τ j usually decreases during the iteration. The pyrosequencing noise removal therefore has two parameters σ p and c p , for all the results presented here these were set at the values 1/60 and 0.01 respectively.
Removing PCR Noise
This requires alignment of the read to the sequence. Alignment was performed with a specially modified version of the Needleman-Wunsch algorithm with a reduced gap cost for homopolymer insertion and deletions. This accounted for the possibility of pyrosequencing noise on low frequency reads which may not have been removed in the flowgram clustering. Gap penalties were included in the distance measure.
PCR per base error probabilities for the three 'Even' V2 'Mock Communities'
Nucleotide | A | C | T | G | Total f | Per cycle p |
A | 0.9995 | 7.2e-6 | 7.7e-6 | 5.1e-4 | 5.2e-4 | 3.5e-5 |
C | 1.1e-05 | 0.9996 | 4.1e-4 | 2.1e-6 | 4.2e-4 | 2.8e-5 |
T | 9.0e-6 | 5.7e-4 | 0.9994 | 1.4e-5 | 5.9e-4 | 4.0e-5 |
G | 3.5e-4 | 3.2e-6 | 2.1e-5 | 0.9996 | 3.7e-4 | 2.5e-5 |
We used a mixture model to cluster the sequences, just as for pyrosequencing noise removal, where each component of the mixture corresponds to a true sequence about which observed noisy reads are distributed. The relative weights of each component are the true relative frequencies of the sequences. The reads are assumed to be distributed as exponentially decaying functions of their sequence error corrected distance from these true sequences. The magnitude of the sequence noise is described by the characteristic length of these exponentials, σ s . A maximum likelihood fit of the mixture model can be obtained using an Expectation-Maximization algorithm initialised using the clusters formed from a hierarchical clustering of sequences at a given distance cut-off, c s . In this study we used parameter values of σ s = 0.033 and c s = 0.08, parameters that experience has taught us work well for GS FLX data. For the Titanium data we compared two different values for σ s , 0.1 and 0.04, whilst keeping c s = 0.08. A standard gap was given a penalty of 15.0 and a homopolymer gap, 4.0. Prior to our sequence clustering step we truncated at 220 bp for GS FLX and 400 bp for Titanium because of the increase in error rates at the ends of reads.
Chimera Identification
Chimeras are generated when incomplete extension occurs in one round of PCR and then the resulting sequence fragment acts as a primer for a different sequence in the next round. Consequently chimeras are composed of two (or occasionally more) true sequences with a discrete break point where the transition from one sequence to another occurs. For our nine test data sets we were therefore able to determine which sequences after denoising were likely chimeras by aligning each sequence against the known reference sequences and finding the putative parents and break point which gave the closest match to the query sequence. If the closest match to a chimera of two sequences was at least three nucleotides or better than that to a single reference sequence then the query was considered as a possible two sequence chimera or 'Bimera'. If it was not then it was considered a 'Good' sequence. Similarly if the match was further improved by three nucleotides when two break points were allowed then it was classified as possibly comprised of three sequences a 'Trimera', and again for the transition to a composite of four sequences or 'Quadramera'. However, the sequence was only classified to these putative definitions if the absolute match was sufficiently good as measured in terms of the sequence error corrected distance (e < 0.15 a distance corresponding to one non-homopolymer gap per 100 nucleotides). Otherwise the sequence was denoted as 'unclassified'. These could include contaminants, real unidentified 16S rRNA operons, gross pyrosequencing or PCR errors, or most likely a chimera that failed to fall under our rather strict definition.
Perseus: Chimera Removal as a Problem in 'Supervised Learning'
For real pyrosequencing data we will not know a priori what sequences should be present and therefore chimera identification algorithms are necessary. Given the mechanics of PCR amplification, any chimeras generated will experience at least one less PCR cycle than either parent, consequently both parents of a chimera will be present in the data set and with a frequency at least equal to the chimera. This ignores the possibility of the chimera experiencing preferential PCR bias over its parents, but it will be true in the vast majority of cases. To exploit this observation we developed an algorithm 'Perseus' that considers each sequence in turn and performs exact pairwise alignments to all sequences with equal or greater abundance, the set of possible parents. The two parents and break point that give the closest match to the query are then identified and a three way alignment of these sequences is generated using the mafft-linsi program [18]. We calculate two quantities from this alignment - the first is the PCR error corrected distance from the query to the optimum chimera. For a sequence to be classified as chimeric this distance has to be absolutely smaller than 0.15 and smaller than or equal to the distance to the closest sequence amongst the best possible parents. This simply ensures that the hypothesis that the sequence is a chimera is possible. However, we still have to account for the possibility that the chimeric pattern could have evolved. We do this by calculating a second quantity using the alignment the 'chimera index' I .
Denoting the query sequence , the closest matching parent , and the more distant parent , we calculate using parsimony the sequence ancestral to all three. We find the number of base pair changes along the three branches to , , and denote these x, y, and z respectively. We resolve changes to the two parts of alignment, either the part of the chimera matching parent or parent , and denote these x A and x B , y A and y B , and z A and z B . For a given chimera to be observed, two independent events must occur, changes to the distant parent must occur on that part of the alignment matching . Assuming all base changes are equally likely then the distribution of changes across the two parts will be binomially distributed with probability proportional to the size of each part. Therefore, we can calculate the probability of the changes being as biased or more so than were observed. The same arguments apply for the changes to the closer parent, they should all lie in the part matching the more distant and we can calculate that probability. We then multiply the two probabilities together and take the negative log to obtain an index that will increase the less likely a chimeric pattern is to have evolved. This index is defined for two parent chimeras, our so called 'bimeras', it could be extended to higher order chimeras but we did not do this, finding that it sufficed for identifying these anyway.
We also added to our training data set the result of taking the reference and calculating their indices without regard to sequence frequency: i.e., comparing all sequences to all others. We then used the test data sets with uneven abundances, the V5 'Artificial Community' and the three V2 'Uneven' data sets, for validation. Running them through our algorithm and then using the logistic regressions to generate probabilities of each sequence being chimeric. Those sequences that do not have a good chimeric match have this probability set to zero. We then defined all sequences with a probability of greater than 50% of being chimeric as chimeras. This will minimise total misclassifications [19]. We also trained the classifier with the Titanium V4-V5 data and associated reference sequences but in this case we lacked a separate data set for validation.
Results and Discussion
In addition to running the AmpliconNoise pipeline we also denoised the data sets with the DeNoiser algorithm [9] and using single-linkage preclustering (SLP) at the recommended 2% sequence difference as well as at 1% for comparative purposes [10]. We truncated the reads at 220 bp and 400 bp for GS FLX and Titanium respectively before calculating exact pairwise sequence distances for the SLP algorithm. For SLP we used the same filtered reads as for AmpliconNoise but this was not possible for the DeNoiser since there filtering is through the QIIME pipeline [20]. The QIIME filtering is slightly less stringent than the procedure described above for GS FLX data but more so for Titanium where a quality window is recommended. The read numbers following QIIME filtering are also given in Table 1.
Per base error rates following noise removal
Percentage per base error rates in pyrosequencing reads before and after application of denoising algorithms
Name | Raw | AmpliconNoise | SLP 1% | SLP 2% | DeNoiser |
Divergent | 0.4519% | 0.1877% | 0.8030% | 0.7960% | 0.1987% |
Artificial | 0.3678% | 0.2469% | 1.1733% | 1.1891% | 0.5605% |
Even1 | 0.8767% | 0.6381% | 1.6990% | 2.3572% | 0.7585% |
Even2 | 0.8870% | 0.6456% | 1.7906% | 2.1536% | 0.7986% |
Even3 | 0.8446% | 0.5935% | 1.9895% | 2.4594% | 0.8033% |
Uneven1 | 1.1852% | 0.8998% | 1.9687% | 1.7756% | 1.0897% |
Uneven2 | 0.6713% | 0.4851% | 1.7549% | 2.6078% | 0.6646% |
Uneven3 | 0.6560% | 0.4344% | 1.3370% | 2.0822% | 0.4913% |
Titanium - σ s = 0.1 | 0.4576% | 0.1427% | 0.5830% | 0.7012% | 0.3793% |
Titanium - σ s = 0.04 | 0.1325% |
Noise removal by all the algorithms can be considered a form of mapping. We map a noisy read onto another that we believe really generated that read. To calculate per base error rates after noise removal we must account for the possibility that the mapping may be to the wrong read. To allow for this we estimated the denoised error rates by, for each read, calculating the number of differences between the denoised read it maps to and the closest matching reference of the original undenoised read. The total of the differences across the data set was then normalised by total alignment length to estimate the per base error rate. The results for the four algorithms are given in Table 3. For the DeNoiser results we only used those reads that were included in the AmpliconNoise and SLP data sets ensuring a fair comparison despite the slight differences in filtering. What is immediately apparent is that SLP at both cut-offs does not actually remove errors instead it inflates them. This is due to the high rate of misassignment where a read is mapped not onto the reference that generated it but to a similar but incorrect sequence. The DeNoiser algorithm does better reducing per base error rates in most cases but it is substantially out-performed by AmpliconNoise which is capable of reducing noise by one-third to a half in all data sets. Given that some residual error will always remain because the sequencing of the references may not be entirely accurate and because of PCR chimeras and contaminants then this is impressive.
Relative Importance of Pyrosequencing and PCR noise
Chimera Frequencies
The classification of denoised sequences from the nine test data sets
Name | Unique | Good | Bimera | Trimera | Quadramera | Unclassified |
Divergent | ||||||
AmpliconNoise | 79 | 56(70.9%) | 21(26.6%) | 1(1.3%) | 0 (0.0%) | 1(1.3%) |
DeNoiser | 37 | 28 (75.7%) | 7 (18.9%) | 1 (2.7%) | 0 (0.0) | 1(2.7%) |
Artificial | ||||||
AmpliconNoise | 118 | 94(79.7%) | 21(17.8%) | 0 (0.0%) | 0 (0.0%) | 3(2.5%) |
DeNoiser | 59 | 46 (78.0%) | 8 (13.6%) | 0 (0.0%) | 0 (0.0%) | 5(8.5%) |
Even1 | ||||||
AmpliconNoise | 2341 | 89 (3.8%) | 1847 (78.9%) | 244 (10.4%) | 4 (0.2%) | 157(6.7%) |
DeNoiser | 289 | 63 (21.8%) | 161 (55.7%) | 36 (12.5%) | 3 (1.0%) | 26(9.0%) |
Even2 | ||||||
AmpliconNoise | 2082 | 90 (4.3%) | 1651 (79.3%) | 227 (10.9%) | 4 (0.2%) | 110(5.3%) |
DeNoiser | 285 | 64 (22.5%) | 171 (60.0%) | 22 (7.7%) | 4 (1.4%) | 24(8.4%) |
Even3 | ||||||
AmpliconNoise | 2210 | 91 (4.1%) | 1781 (80.6%) | 188 (8.5%) | 3 (0.1%) | 147(6.7%) |
Denoiser | 287 | 64 (22.3%) | 170 (59.2%) | 24 (8.4%) | 0 (0.0%) | 29(10.1%) |
Uneven1 | ||||||
AmpliconNoise | 1124 | 94 (8.4%) | 816 (72.6%) | 81 (7.2%) | 1 (0.1%) | 132(11.7%) |
Denoiser | 212 | 61 (28.8%) | 111 (52.4%) | 9 (4.2%) | 0 (0.0%) | 31(14.6%) |
Uneven2 | ||||||
AmpliconNoise | 859 | 77 (9.0%) | 669 (77.9%) | 71 (8.3%) | 2 (0.2%) | 40(4.7%) |
Denoiser | 154 | 49 (31.8%) | 87 (56.5%) | 9 (5.8%) | 1 (0.6%) | 8(5.2%) |
Uneven3 | ||||||
AmpliconNoise | 1053 | 75 (7.1%) | 843 (80.1%) | 82 (7.8%) | 0 (0.0%) | 53(5.0%) |
Denoiser | 202 | 55 (27.2%) | 124 (61.4%) | 7 (3.5%) | 0 (0.0%) | 16(7.9%) |
Titanium | ||||||
AmpliconNoise - σ s = 0.1 | 163 | 76 (46.6%) | 77 (47.2%) | 1 (0.6%) | 0 (0.0%) | 9(5.5%) |
AmpliconNoise - σ s = 0.04 | 304 | 91 (29.9%) | 174 (57.2%) | 2 (0.7%) | 0 (0.0%) | 37(12.2%) |
DeNoiser | 151 | 14 (9.3%) | 70 (46.4%) | 6(4.0%) | 0 (0.0%) | 61(40.4%) |
Accuracy of OTU Construction Following Noise Removal
Run times in seconds for AmpliconNoise and the original PyroNoise for the V5 'Artificial Community' data set
AmpliconNoise | PyroNoise | |||||
Process | Cores | Wall | CPU | Cores | Wall | CPU |
Initial pairwise distances (NDist) | 64 | 1900 | 45358 | 64 | 1900 | 45358 |
Initial clustering (FCluster) | 1 | 292 | 252 | 1 | 292 | 252 |
Flowgram clustering (PyroNoise) | 64 | 1904 | 9364 | 64 | 55241 | 686497 |
Sequence clustering (SeqNoise) | 64 | 503 | 20713 | -- | -- | -- |
Total | 4599 | 75687 | 57433 | 732107 |
In Figure 3 we also show the effect of applying single-linkage preclustering at 1% and the recommended 2% level, and the DeNoiser algorithm prior to OTU construction. Single-linkage preclustering at 1% is very poor greatly over-estimating OTU numbers, 2% SLP and the DeNoiser perform similarly above a 3% cut-off predicting slightly more OTUs than the original PyroNoise program, both were worse than AmpliconNoise. Below this 2% SLP predicts a constant OTU number suggesting it is aggregating OTUs that should be separated. However, both SLP and DeNoiser were orders of magnitude faster than AmpliconNoise, single-linkage preclustering does not require a cluster and for this data set it ran in just a few minutes on a standard computer. The DeNoiser took less than an hour on a single core.
Accuracy of 3% OTU construction following application of the noise removal algorithms
Name | AmpliconNoise | SLP 1% | SLP 2% | DeNoiser | ||||||||
3% OTUs | Good | Missed | Noise | Good | Missed | Noise | Good | Missed | Noise | Good | Missed | Noise |
Divergent | 26 | 0 | 1 | 26 | 1 | 162 | 26 | 1 | 11 | 24 | 0 | 3 |
Artificial | 34 | 0 | 1 | 38 | 0 | 109 | 32 | 2 | 10 | 36 | 0 | 6 |
Even1 | 57 | 0 | 131 | 60 | 2 | 174 | 39 | 19 | 65 | 59 | 1 | 21 |
Even2 | 57 | 0 | 89 | 59 | 2 | 137 | 42 | 15 | 53 | 58 | 3 | 22 |
Even3 | 58 | 0 | 125 | 65 | 0 | 197 | 43 | 17 | 75 | 58 | 3 | 23 |
Uneven1 | 58 | 0 | 101 | 60 | 0 | 144 | 47 | 10 | 59 | 54 | 7 | 24 |
Uneven2 | 48 | 0 | 35 | 51 | 0 | 74 | 38 | 10 | 26 | 46 | 6 | 6 |
Uneven3 | 49 | 1 | 46 | 51 | 2 | 97 | 41 | 10 | 45 | 48 | 6 | 12 |
Titanium | ||||||||||||
σ s = 0.1 | 66 | 0 | 8 | 74 | 0 | 169 | 67 | 1 | 18 | 15 | 53 | 14 |
σ s = 0.04 | 67 | 0 | 21 |
Chimera Classification Accuracy
Chimera classification accuracies Perseus applied to the three denoised V2 'Uneven' data sets.
Dataset | Uneven1 | Uneven2 | Uneven3 | |||
Classification | Good | Chimeric | Good | Chimeric | Good | Chimeric |
Good | 78 (83.0%) | 16 (17.0%) | 70 (90.9%) | 7 (9.1%) | 62 (82.7%) | 13 (17.3%) |
Bimera | 7 (0.9%) | 809 (99.1%) | 9 (1.3%) | 660 (98.7%) | 10 (1.2%) | 833 (98.8%) |
Trimera | 1 (1.2%) | 80 (98.8%) | 1 (1.4%) | 70 (98.6%) | 1 (1.2%) | 81 (98.8%) |
Quadramera | 0 (0.0%) | 1 (100.0%) | (0.0%) | 2 (100.0%) | -- | -- |
Unclassified | 26 (19.7%) | 106 (80.3%) | 14 (35.0%) | 26 (65.0%) | 22 (41.5%) | 31 (58.5%) |
Chimera classification accuracies for ChimeraSlayer applied to the three denoised V2 'Uneven' data sets.
Dataset | Uneven1 | Uneven2 | Uneven3 | |||
Classification | Good | Chimeric | Good | Chimeric | Good | Chimeric |
Good | 86 (91.5%) | 8 (8.5%) | 72 (93.5%) | 5 (6.5%) | 72 (96.0%) | 3 (4.0%) |
Bimera | 125 (15.3%) | 688 (84.3%) | 98 (14.6%) | 571 (85.4%) | 108 (12.8%) | 735 (87.2%) |
Trimera | 20 (24.7%) | 61 (75.3%) | 13 (18.3%) | 58 (81.7%) | 15 (18.3%) | 67 (81.7%) |
Quadramera | 0(0.0%) | 1 (100.0%) | (0.0%) | 1 (50.0%) | -- | -- |
Unclassified | 55 (41.7%) | 76 (57.6%) | 15 (37.5%) | 26 (62.5%) | 27 (50.9%) | 24 (45.3%) |
In practice, dedicated training data sets for each study may not be possible, although we would recommend it. If data is GS FLX V5 or V4-V5 Titanium then the two pairs of α and β values given above should work well for their corresponding data types. For V2 data, we would not recommend the values in the caption of Figure 8 because there is an implicit assumption of very high prior chimera probability generated from training on this atypical data set, reflected in the high value of α = -2.83542. The β values on all data sets are in the range of β = 0.5 and we have found that this value, paired with α = -7.5, performs well across a wide range of data sets.
We have demonstrated that AmpliconNoise followed by Perseus has the sensitivity to remove the majority of errors from GS FLX and Titanium pyrosequenced amplicons and allow accurate estimates of OTU number. AmpliconNoise outperforms both agglomerative clusterers, SLP and the DeNoiser, both in terms of per base error rates and OTU construction but at a cost of increased computational complexity and no doubt in some cases, where some noise can be tolerated, these simpler heuristic approaches may be the best option. However, the results here suggest that both agglomerative approaches must be treated with caution, by not simply looking at OTU numbers as in previous evaluations [9, 10], but rather their identity we have established that these are prone to over-clustering, removing a substantial fraction of the true diversity even at the 3% level.
Consequently, we believe that the AmpliconNoise-Perseus pipeline which is freely available as open source software [23] with all data on a dedicated website [24], will find a wide range of applications in microbial diversity estimation [8], and population genetics [2]. They could be critical to the success of large-scale publicly funded efforts to explore microbial diversity, such as the sequencing of human associated microbes being conducted in the Human Microbiome Project [25]. To facilitate their use we include programs for integrating their output into the QIIME 16S rRNA analysis pipeline [20].
We have shown the importance of considering both the effects of PCR and sequencing errors in studies of diversity based on 16S rRNA amplicons. This suggests the use of high fidelity polymerases to reduce per base PCR error rates. However, one of our most striking observations was just how variable chimera frequencies were in the test data sets. This must be due to PCR conditions, principally, cycle number, extension time, primer and template concentrations and polymerase type [26]. Therefore optimising the whole PCR process to minimise all types of errors is probably a better strategy than just focussing on the enzyme.
In addition, the principles outlined here: the rigorous validation of noise removal algorithms with test data; the use of EM algorithms to generate effective consensus sequences and remove noise from pyrosequenced amplicons; using sequence abundances in the classification of PCR chimeras; and the treatment of the latter as a supervised learning problem; will provide the basis for further algorithm development in this field and contribute to the maturation of next generation sequencing as a quantitative technique for the analysis of PCR amplicon diversity. These principles will be equally applicable to other pyrosequencing platforms, for example the Illumina HiSeq 2000 which is capable of generating orders of magnitude more reads per run [27, 28]. They should even hold for the third generation of sequencing technologies that will target individual molecules [29]. As ever larger amounts of sequence data are generated the question of how to distinguish true diversity from noise will become ever more important.
The Titanium full length clones sequences have been submitted to GenBank with accession numbers HQ462473-560. The Titanium pyrosequencing reads have been submitted to the Short Read Archive with accession SRP003773.
CQ is supported by an Engineering and Physical Sciences Research Council Career Acceleration Fellowship (EP/H003851/1). PT is supported by NIH P50 GM068763. We would like to thank Jens Reeder and Rob Knight for providing the DeNoiser processed data sets and Sue Huse for a copy of the single-linkage preclustering algorithm. We also thank two anonymous reviewers and Dr Keith Harris for constructive comments on the manuscript.
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This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


text:
Altitude and Alcohol - what a mix!
Trip Start
Jun 13, 2006
Trip End
Jun 12, 2007
Cusco is very pretty and colonial with a few amazing Incan walls thrown in for good measure - how they fitted them so well I will never know. It is also the most touristy place we have been since we started this trip. On the Plaza de Armas (the main square) and for about 4 blocks in every direction around it all the shops are tourist related: tourist restaurants, bars, tour operators, internet cafes, native craft shops etc... and as you walk down the street people try to entice you into their establishment or try to get you to buy their crafts, paintings, have a shoe-shine (even if you are wearing trainers - albeit mucky), it is quite exhausting after a while! That said we have done quite a bit of shopping mainly buying practical things for the Inca Trail, or at least things that I want and claim we need for the Inca Trail (see the photos for some of our more attractive purchases).
One of the things we did discover is that alcohol and altitude really do not mix. We popped into a bar for a quick drink at about 6 and stayed through 2 happy hours before heading home sometime after 12 - opps. The bar claims to be the highest Irish owned pub in the world (with another Irish pub round the corner a slightly dubious claim I think). Anyway we met many new friends and had a lot of fun (and probably made fools out of ourselves) but have some definite memory lapses and wasted a whole day with the hangovers from hell! First G&Ts of the trip though, yum.
Inca Trail tomorrow...4 days of walking at over 4000m above sea level - is this a good idea?


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Centrepiece - Hamilton, ON
in Abstract Public Sculptures
A newer sculpture at the Hamilton Art Gallery
posted by: stinger503
location: Ontario, Canada
date approved: 4/21/2012
last visited: 12/17/2014
N0 km
Hollow Form with Inner Form - Hamilton, ON
Small sculpture in sculpture garden
date approved: 4/19/2012
S0 km
Breath - Hamilton, ON
Piece of art outside Hamilton art museum
W0 km
Portal - Hamilton, ON
Two piece sculpture in Hamilton
SE0 km
Canadac - Hamilton, ON
Large sculpture outside art museum
E1 km
Thespis - Hamilton, ON
Piece of art in Hamilton
date approved: 5/1/2012
last visited: 4/14/2013
N2.4 km
Sculpture of "Rafaga - Unleashed"
Located beside Parks Canada's Marine Discovery Centre on the Waterfront Trail.
posted by: The Blue Quasar
date approved: 4/18/2007
last visited: 6/29/2013
W35.4 km
Meg's Pause - Glenhyrst Gardens, Brantford, Ontario
This fifteen-foot, two-dimensional drawing of hands in steel, is located in the Glenhyrst sculpture gardens.
posted by: 13coach13
date approved: 7/29/2010
last visited: 6/4/2012
NW42 km
Tribute to Nahneebahweequay by Tom Benner
The Queen Victoria sculpture is my favourite sculpture in the Arboretum at the University of Guelph. Queen Victoria resides high over the Centre, facing Nahneebahweequay who stands inside the Centre looking out at Queen Victoria.
posted by: J.A.R.S.
date approved: 8/6/2006
last visited: 8/31/2013
NW42.3 km
Mask by Evan Penny - University of Guelph
Evan Penny Mask, 1989 bronze Donald Forster Sculpture Garden, Macdonald Stewart Art Centre University of Guelph
date approved: 8/3/2006
last visited: 4/8/2016
Agricultura - Guelph, Ontario, Canada
A sculpture located in the Donald Forster Sculpture Park in Guelph, Ontario.
posted by: monkeys4ever
date approved: 12/2/2009
NW43.3 km
Garbasaurus - Guelph, Ontario
Located in Royal City Park next to the Speed River near Gordon Street. Artist unknown.
date approved: 8/7/2006
N48.6 km
Green Door - Peel Heritage Complex - Brampton, Ontario
The curved green door outside the Peel Heritage Complex
posted by: northernpenguin
date approved: 9/16/2008
last visited: 8/8/2010
NE51.2 km
Tilted Spheres - Toronto Pearson International Airport - Mississauga, Ontario.
Tilted Spheres is a large piece located in terminal 1 at Toronto Pearson.
posted by: saopaulo1
date approved: 3/16/2010
last visited: 5/2/2013
E51.8 km
Abstract Public Sculpture - "Circle of Friends", St. Catharines City Hall, ON
Created by a local artist, this piece has also been on display in Hamilton and in New York State's "Art Park"
date approved: 6/2/2008
last visited: 11/5/2010
E52 km
Abstract Public Sculpture - "Untitled", St. Catharines Public Library
This unnamed piece is on permanent display inside the St. Catharines Public Libary on the first floor.
date approved: 7/31/2008
last visited: 1/18/2014
Abstract Public Sculpture - "Tribute to Women", St. Catharines Public Library
Featured artwork near the main entrance to the St. Catharines Public Library.
last visited: 6/25/2016
E52.8 km
APS - Path of Possibilities
Located near the main entrance to the Brock Tower on the university campus.
date approved: 7/23/2007
last visited: 7/5/2012
E52.9 km
APS - She Wolf
An amazing piece in the centre of the traffic circle on the campus of Brock University.
last visited: 7/19/2012
NE54.9 km
The Temple - Toronto, Ontario, Canada
This sculpture is located in High Park in Toronto.
date approved: 12/27/2009
last visited: 5/15/2009
NE55.4 km
Time Capsule - Toronto, ON
A Kosso Eloul sculpture near Ontario Place
date approved: 8/2/2016
last visited: never
E56.2 km
APS - "Carmpounded" by Mykl Hulley
Mounted to the exterior wall of the Niagara Gallery.
date approved: 7/14/2007
last visited: 6/7/2008
NE57.6 km
Flower Power - Toronto, ON
This restored red sculpture has been relocated here from its former home in High Park.
posted by: ras258
date approved: 10/9/2011
Barca Volante - Toronto, ON
Barca volante flies over a rectangular lake just west and south of Rogers Centre.
date approved: 9/21/2011
NE57.9 km
Spiral Fountain - Toronto, ON
This sculpture is in the garden in front of the Renaissance Hotel.
last visited: 5/31/2012


text:
Vegan Stir Fry Recipe
Just a basic , simple stir-fry recipe I got from my aunt. I cooked this for my Mom one night and she loved it. If you're not vegetarian, it is really easy...
asparagus, cabbage, broccoli, rice, tofu, mushroom, red pepper, sesame seed, +garlic, +soy sauce, +ginger, +olive oil, +onions, +zucchini
www.grouprecipes.com - Similiar recipes
Eriks Teriyaki Tofu Stirfry
tofu, broccoli, cabbage, mushroom, rice, sesame seed, +bamboo shoots, +baby carrot, +vegetable oil, +teriyaki sauce, +sugar
vegweb.com - Similiar recipes
Brown rice chicken
rice, red onions, mushroom, broccoli, red pepper, sesame seed, +water, +chicken broth, +chicken, +olive oil, +garlic, +lemon, +white wine, +green olive, +cheese, +bread crumbs
www.nibbledish.com - Similiar recipes
Sesame Vegetable Tofu Stir-Fry
tofu, broccoli, red pepper, cabbage, sesame seed, rice, +soy sauce, +ginger, +sugar, +dark sesame oil, +garlic, +vegetable oil, +bean sprouts
find.myrecipes.com - Similiar recipes
Fennel Vegetable Stir Fry with Nut Rice
cabbage, red pepper, broccoli, mushroom, tofu, rice, +almond oil, +yellow onions, +carrot, +fennel seed, +almonds, +salt, +black pepper, +garlic
vegweb.com - Similiar recipes
Thai Chicken Stir Fry Recipe
THAI CHICKEN STIR FRYThis recipe came from an estate sale. I obtained it when I purchased the family collection from the Brink Estate in Big Spring, Texas...
broccoli, sesame seed, rice, mushroom, red pepper, red onions, +chicken, +carrot, +chicken broth, +chili oil, +green pepper, +green onion, +peanut sauce, +red chili pepper, +peanut butter, +water
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Barbecue Ginger Marinated Vegetables Recipe
This is one of the many varaiated vegetable side dish i like to do for my BBQ'S.. Prep:15m
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Stir Fried Sesame Vegetables with Rice
"In this dish, the peanut oil is preferable to the vegetable oil, as the flavor of the peanut oil adds a really nice nutty flavor."
asparagus, rice, mushroom, red pepper, sesame seed, +garlic, +margarine, +vegetable oil, +soy sauce, +vegetable broth, +yellow onions
allrecipes.com - Similiar recipes
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Legal & General will face intense scrutiny this week, as analysts and investors, both increasingly worried by the prospect of yet more turmoil in the equity markets and the possibility of higher-than-expected corporate bond defaults, focus on the strength of the insurer's capital base.
L&G, which is due to post full-year numbers on Wednesday, has already issued an update on its funding at the end of last year, telling the market last month that its regulatory capital surplus stood at more than £1.6bn as at 31 December. But worries persist, not least, as the stockbroker Charles Stanley highlighted in a note to clients last week, because of the numerous differing accounting measures that investors must grapple with when valuing life insurance stocks in the current environment.
The dividend is likely to be a key area of focus, with some in the market arguing that a reduction of the payout may be appropriate in light of the current market turmoil. MF Global, for example, does not anticipate the need for a recapitalisation, but warned that a dividend cut and a scaling back of new business may be the best options on the table.
"Given that the company raised capital at the bottom of the last equity market downturn, one can certainly see why the market is concerned," the broker said.
"However, now is most definitely not the time to raise equity if you do not need to, and we see it as far more likely that the company will simply adjust its business model and dividend to the current circumstances."
Today: Xaar, the print technology group, is due to publish its preliminary results this morning and, according to the broker Panmure Gordon, "unless the company pulls something amazing out of the bag", the share price, which has rallied very strongly over the past three months, is unlikely to climb significantly higher.
The headline figures have already been flagged, with full-year profits and revenues falling by around 12 per cent and 40 per cent respectively.
"We like the technology and the company's position in the market, but believe the macro conditions will remain tough for some time," Panmure said in a note to clients last week.
"Recent product launches should help to reduce pricing pressures but will not offset the macro headwinds."
Also today, Ceres Power, the Alternative Investment Market-listed fuel cell specialist, is due to post interim results and updates on development milestones that are more important than the actual figures, as the company is not expected to make a profit until 2014.
Results/updates: Daily Mail & General Trust, Huntsworth, Ceres Power, Xaar.
Tomorrow: Results/updates: Motivcom, Sportech, Bioquell, Severn Trent, Ferrexpo, Renewable Energy Generation, Carnival.
wednesday: J Sainsbury is due to post a fourth-quarter trading update. Citigroup expects a 6.5 per cent hike in ex-fuel like-for-like (LFL) sales, which would be the supermarket group's best performance in three years. The broker highlights that its forecasts factor in the around 90 basis points impact of the new VAT rate, and are adjusted for a later Easter.
"So far, we only have TNS market share data covering the first seven weeks of the 11-week period – and the next set of data we receive will not come out until after these Sainsbury figures," Citi said.
"So TNS is only a partial guide, but it suggests that Sainsbury has made a very solid start to the year, with growth of over 7 per cent in both of the last two periods. For this reason, we expect [the company] to report better LFL sales growth than the 4.9 per cent it reported for its third quarter."
Results/updates: Smiths, TUI Travel, Henry Boot, Eurasian Natural Resources Corporation, Hochschild Mining, Imperial Tobacco, Melrose Resources, Nichols, Salamander Energy, Vislink, Pinewood-Shepperton, J Sainsbury, Legal & General.
Thursday: The advertising agency M&C Saatchi is due to publish preliminary results. Numis Securities forecasts £14m in pre-tax profits, with 14.9p in earnings per share.
Saatchi is expected to follow in the steps of the large advertising holding groups, several of whom have already reported a deterioration in trading in the fourth quarter of last year and warned of a difficult start to 2009.
Results/updates: Abbey Protection, Kingfisher, Premier Oil, PV Crystalox, United Utilities, M&C Saatchi, Northern Foods.
friday: Results/updates: Clinton Cards, Dairy Crest, Trading Emissions, JKX Oil & Gas, Just Retirement.
- More about:
- Panmure
- Retirement
- Sainsbury (j)
- Severn Trent


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Subject: Posted: October 31, 2012 01:44 pm
Take away one barkgal and add one more blaster blade.
take away two Covenant Knight, Randolf and add two more Little Sage, Marron. add two more Solitary Knight, Gancelot or take out gancelot and add 3 soul saver dragon. add more Starlight Unicorn for the power boost. also add more grade 3's so you have more chances of drawing them. I go by this rule: 17 grade 0's (16 triggers + starter vanguard) 12 grade 1's 12 grade 2's 9 grade 3's but these are suggestions,the choice is up to you |
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vegitto96 |
Subject: my deck Posted: October 31, 2012 02:25 pm
thanks for the good advise silent i will keep it in consederation
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Marketing managers are forecasting increased sales growth over the next 12 months. They predict sales volume growth of 6.3 per cent compared to 5.4 per cent in the previous year. But prices are also expected to increase by 2.1 per cent, compared with planned rises of 2.5 per cent reported three months ago. The forecasts emerged in the latest Marketing Trends Survey by The Chartered Institute of Marketing. CIM officials say the reduction in the planned rate of price increase is an indicator that the upsurge in inflation caused by the weak pound and last year's price boom could be over. "This is especially important since the Governor of the Bank of England has been arguing that the risk of accelerating inflation pointed to the need to raise interest rates," said a spokesman.
- More about:
- Bank Of England
- Inflation
- Marketing
- Sales


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The Iphone has sold about 11,000 phones in India. For all the hype, thats a mighty small figure.
The gray market for the old, 2G Iphone (pity they killed it) is much larger, I will bet on that...
But, we had foreseen it...
Saturday, November 15, 2008
I told you so: Iphone edition
1 comment:
- Kavi said...
The grey market in India opened up a new market for people who break lock-ins !!
- 1:02 PM, November 15, 2008
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I tried these at a restaurant this weekend and I am sharing them with you here today. This can only mean one thing. I absolutely loved them. I loved them so much that despite the fact that I had them for dinner on Saturday night, I rushed out to the store, bought the ingredients and made them for dinner on Sunday night. They really were that good. And with Christmas less than a week a way now, it might be a good idea to run your grocery store, grab all the ingredients and add these mashed potatoes filled with green onions and succulent dungeness crab meat to your holiday menu. It will be like giving your taste buds a Christmas present.
Dungeness crab is much loved here in the Pacific Northwest, you can find it just about anywhere. From Pike's Place Market to Costco, to the small market just around the corner. It is everywhere. And I'm going to let you in on a little secret: it's sweet delicate flavor makes me like it more, much more actually, than its more expensive counterpart, king crab. Sit me down in a front of a plate of hot steamed dungeness crab and I am one happy girl. But add that same delicious crab to my all time favorite comfort food, mashed potatoes, and I am down right giddy.
While I do recommend dungeness crab with this recipe, I do realize that not everyone has access to it. If you do not, just use whatever kind of fresh lump crab meat you can find. Or if you really want to fancy things up, how about lobster? Enjoy!
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You are in: Sci/Tech | ||||||||||||||||||||
Wednesday, 26 December, 2001, 08:56 GMT
Expedition to the lost net
The global reach of the net is one of its greatest charms.
But there are some parts of the net that no-one can reach, parts that are cut off from even the most dedicated net user either by accident or design. A study of these lost sites by US researchers has investigated the extent of the "dark net" and just what causes a net address to fall off the map. The study found that up to 5% of the net - potentially 100 million hosts - is completely unreachable. Return to sender The internet works because all the computers on it have an individual address. Routers, devices that shuffle data around the net, know where to send information because they maintain lists of these addresses. But occasionally some of these addresses disappear and researchers at Arbor Networks have spent three years finding out why. The researchers found that the number of sites an individual surfer can see depends on their starting point. Contractual wrangles between net service providers can mean that customers of the rowing firms are cut off from portions of the web that the rival firm gives access to.
The study reveals that all parts of the net are not equally connected to every other part. By contrast, some networks inadvertently act as gatekeepers for sections of the net that lie beyond them. Hijacked routers In their study, the three researchers, Craig Labovitz, Abha Ahuja and Michael Bailey, reported that many of the lost net sites flare into life briefly when being used to send spam (unsolicited commercial e-mail messages) or to launch attacks on other parts of the web. The Computer Emergency Response Team, which monitors security problems online, has already issued a warning about malicious hackers hijacking routers to use as a launch pad for attacks. The specifications that govern the way routers co-ordinate to ensure the net keeps running are unfortunately susceptible to subversion by those who are determined enough. Routers can be made to pose as particular net addresses and be used to launch barrages of data at other target sites. "The sheer quantity of routing information, coupled with the lack of security both in routers and the routing protocol itself create an infrastructure that is increasingly unwieldy and vulnerable," said Craig Labovitz, lead researcher on the Arbor Network study. Other parts of the internet, mainly sites used by the US military, are lost because they use old addresses that no router references anymore. |
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Michael Owen is once again playing no part in a vital 2010 World Cup qualifier but his absence is doing a wonderful job of focusing the minds of those who have left him behind. Rio Ferdinand admitted yesterday that even England's top players now recognise the stark truth that, under Fabio Capello, reputations count for nothing.
Capello has demonstrated to his England players there will be no room for sentiment with his treatment of Owen, whom he has overlooked this week despite injuries to Emile Heskey, Carlton Cole and Darren Bent. Ferdinand said yesterday that the players now understand that – whatever their status – they cannot afford to lose either form or fitness under Capello if they want to stay part of his plans.
Ferdinand is set to reclaim his place tonight against Ukraine, in place of Matthew Upson, after sitting out Saturday's 4-0 win over Slovakia with a groin injury. Ferdinand said: "There can be no complacency in this squad. Look at Michael Owen – he's got 40 goals in 89 appearances. You'd have said he was bang on to get 100 caps two or three years ago. But this manager has come in and he's seeing things from a different angle."
The Manchester United defender said he believed the Capello approach, and the abolition of favouritism, was something the England team had not experienced for a quarter of a century but that it was exactly what was required. Ferdinand said: "Over the last 20 or 25 years there have been four or five players who you could say it didn't matter if they played poorly. They'd always get the shirt. Now no one is guaranteed to be in the team.
"It's better if people are on their toes. It keeps the hunger in your belly, the desire, the passion. If you're not fit enough, or not on form, you won't get in the squad. That's the way it's got to be. Look at what the manager has won. You don't win what he's won by having a lot of sentiment.
"It's black and white, no grey areas. No ifs, buts or maybes. [Capello says] 'This is what I want from you. If you can't do it, then you're not the right man to be in the squad'."
Ferdinand sampled life on the outside during Saturday's friendly at Wembley, when he watched Upson's accomplished display alongside the captain, John Terry. He admitted that such stiff competition for places made him uneasy. He said: "That was all I was thinking about during the game, that someone else was getting an opportunity to stake their claim."
Terry and Ferdinand remain Capello's preferred choice at the centre of defence, despite a persistent back injury restricting the latter's recent performances for England – they have only played alongside one another four times in the 15 months that Capello has been in charge. The pair last played together in September's pivotal 4-1 victory over Croatia in their World Cup qualifier in Zagreb.
- More about:
- Fabio Capello
- Fitness
- Matthew Upson
- Michael Owen


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- About me
- Cenantua – What and why?
- Cenantua's pages (bio sketches & so on)
- Citing… this site
March 7, 2013 by Robert Moore
As part of the effort to find those elusive Southern Unionists, it shouldn't be surprising that I would look to the activities of the Grand Army of the Republic in the hopes of finding some of the local boys in blue. Indeed, there were a couple of posts in the Shenandoah Valley, and one of […]
December 13, 2012 by Robert Moore
No, not about Southern Unionists or the Valley… but there's a tie to the Valley… just wait for it a bit. In Don Troiani's print, "Bronze Guns and Iron Men", there is an officer other than John Pelham, just behind the Napoleon, with binoculars in hand. To most, it might appear like a section commander… […]
December 2, 2008 by Robert Moore
This is interesting and something of which I had never heard. Thanks to a link from Jenny's Draw the Sword Blog blogroll, I took a little Web-trip to the Cordori family site and found this interesting page about George J. Codori. If you are familiar with the Civil War, the Codori name should quickly ring […]


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"Her on-screen persona was basically sexless. In her most famous films, she was a cartoon, a parody of femininity. Paired with sexless leading men like David Wayne, Tommy Noonan, Tom Ewell, or as in 'Some Like It Hot,' men who were in drag and/or pretending to be impotent, Monroe was all display and no action. Her on-screen relations with the opposite sex were infantile and tentative . . . The legend of Monroe's screen sexuality is more accurately a myth. She was the perfect, pliable non-threatening female figure for the frightened '50s: no anger, no inner life, no control of her own sensuality." --Denis Ferrara writing in Talk magazine's special issue "An Oscar Odyssey, March 2001.
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May. 05, 2014
The Iron & Steel Technology Conference and Exposition features technologies for the steel industry and provides a global perspective on the expertise needed to power a sustainable steel industry. Praxair offers a wide range of technologies and services to the global metals industry including the CoJet™ gas injection system, the argon oxygen decarburization process, stove oxygen technology, and dilute oxygen combustion for steel reheating.
Praxair Booth #432
Indiana Convention Center 100 S Capitol Ave Indianapolis, IN


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John Adrian Tremayne Rodd, rugby player: born 28 June 1935; capped 14 times for Scotland 1958-65; succeeded 1978 as third Baron Rennell; married 1977 Phyllis Neill (one son, three daughters); died London 9 December 2006.
The strong competitive streak that ran through Tremayne Rodd was not confined to the rugby field, on which he won 14 caps for Scotland and played for the Barbarians. The future Lord Rennell was equally combative around the backgammon table and on the golf course.
First capped for Scotland in the 11-9 win over France at Murrayfield in 1958, he had an international career that stretched over seven years as he kept battling his way back into the side. An ever-present in the five Tests played in 1958 - he was on the winning side against Australia and in the team that drew with England in the Calcutta Cup match in Edinburgh - he missed the 1959 campaign and played in the opening two defeats in the 1960 championship.
He was missing for the 1960/61 season and played in the opening game of the 1962 championship against France. Stan Coughtrie overtook him again in 1963 before he returned triumphantly the next season. After playing in the opening success over the French at Murrayfield he helped Scotland draw 0-0 with the touring All Blacks. He then lost his place again, this time to Alex Hastie, after a narrow defeat in Cardiff to Wales. There were three more appearances in 1965, all of which were on losing sides, before his Test career came to an end. By then he was 30, but a long way off from hanging up his boots.
In fact, his last-known appearance on a rugby field was in 1993, at the ripe old age of 58, when he appeared for a House of Lords and Commonwealth Officers XV against a South African President's XV in Cape Town.
"Tremayne was fiercely competitive and always liked to keep himself in shape. He was a very good scrum half when I played with him for London Scottish and Scotland who had a quick pass and a lightning break," recalled Iain Laughland, the former Scottish Rugby Union President:
He was a great character to have in your side and always raised a smile. I remember going to see him play for a House of Lords team against a Scottish Bankers XV on the morning of an international in Edinburgh. He was well into his fifties at the time and came over to me just before kick-off and asked me to look after his false teeth. I eventually put them in a plastic bag and left them at the corner flag.
Rodd learned his rugby at Downside and Britannia Royal Naval College, Dartmouth, before following his father, Commander Gustaf Rodd, into the Navy. He played for the Navy and United Services while serving and was in the Combined Services XV that met the Wallabies in 1957 and the Springboks in 1961. In 1958 he was home fleet boxing champion.
"When he left the Navy he went into investment banking at Morgan Grenfell, but simply hated it. His uncle was the chairman and I told him to stick with it, but he quit after nine months," Laughland said. "In the end he became the director of a sports trophies company in King's Cross."
A strong golfer who used to play against Sam Torrance at Sunningdale, Rodd was an expert backgammon player who competed at the world championships. One Christmas, before he succeeded his uncle as third Baron Rennell in 1978, he set up a table in a window at Selfridges in London where he challenged all-comers to play him.
He met his wife, Phyllis Neill, in a queue at an airport as he was returning from Italy after his father's funeral. He allegedly asked her to hold a bag for him while he went to the lavatory. The bag contained the urn in which his late father's ashes had been placed.
Rob Cole


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Schematics / Magic Smoke LFX-LFO
< Magic Smoke LFO, design by Tim Servo | Schematics | Moog Source VCLFO redrawn by Marjan Urekar > DIY | LFO | LFX | Wave shaping | Magic Smoke | Tim Servo | Triangle Magic Smoke LFX - LFO ,with a waveshaper output ,more on this thread : http://electro-music.com/forum/viewtopic.php?highlight=lfo&t=49081 ,from Tim Servo : Here's a LFO design I've been having a lot of fun with lately. The LFO core is my "8K" design that some of you have seen before. It's a variation on the standard Integrator / Comparator triangle core LFO, and I tweaked it a bit to give it a wider range (about 8,000 : 1, hence the name). The waveshapers are a combination of a tri-to-saw waveshaper ("Wave A" which is basically a sign changer) and my own TSQ tri-to-square waveshaper ("Wave B"). Here's a link to a YouTube vid where I put in a different cap to see (and hear) what the various wave combinations looked like: I call it the LFX. It's a pretty easy circuit to build, and will definitely provide you some very different LFO waveforms. Enjoy!! NOTE: THIS IS AN UPDATED REV 02 VERSION WITH A NEW VALUE FOR R5. < Magic Smoke LFO, design by Tim Servo | Schematics | Moog Source VCLFO redrawn by Marjan Urekar > |
Edit - History - Print - Recent Changes - Search Page last modified on 2013-02-24 17:36 [UTC-7] - 2330 views


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new gluten free buns are awesome. come sealed in bag with burger separate. a nice option for road trips!
warning! I went in and talked to the manager about what items were gluten free. I should have trusted my gut.. but I didn't they told me the chicken strips were gluten free and they had no allergy menu in store. so I purchase them take them home to eat. get glutened look online and Culvers website under the allergen menu says basically everything in it has gluten. so pissed
5750 Ranch Lake Blvd, Bradenton, FL 34202
Always done gf and right !! Have not been sick from these peeps!!
4121 Pioneer Woods Dr, Lincoln, NE 68506
The fries are not gluten free. they are fried in a fryer with breaded items. they have very little gluten free items but do offer a bun if you want a cheeseburger. However, my daughter has gotten sick after eating here 2 separate times. The staff do not seem to care when you mention that you are gluten free. definitely ask for manager when you order.
2530 Woodlawn Rd, Lincoln, IL 62656
Delicious gluten-free buns. Do not have a dedicated fryer. Frozen custard is delicious and can be gluten-free, depending on the toppings.
4400 Wabash Ave, Springfield, IL 62711
Delicious gluten-free buns! No dedicated fryer though. Delicious frozen custard!
632 AZ-92, Sierra Vista, AZ 85635
Only Gluten free knowledge was that they had a gluten free bun. None of my other questions were answered like they knew what was going on.
3604 University Dr, Muscatine, IA 52761
Let me start by saying that I do have celiac disease. I truly enjoy Culver's bacon cheeseburger on a gluten-free bun. I also enjoy their french fries but a word of caution, there could be some cross contamination with the fries. They do have a dedicated fryer for french fries and the workers are trained but mistakes can happen.
7801 Nicollet Ave., Bloomington, MN 55420
I have eaten at this location twice, both times it was great. Burger comes in a box with whatever toppings you order, and then a sealed gluten free bun is in the bag. Some assembly required.
17010 Torrence Ave, Lansing, IL 60438
Very delicious GF warm and soft buns. No dedicated fryer though.
450 Pond Promenade, Chanhassen, MN 55317
Offers GF burger bun, pretty cool! Chili is ok too. and theres frozen custard for dessert. Ah, winning!
Love the gluten free buns on the burgers. Custard too!
3502 W. 29th Street, Greeley, CO 80634
good gf buns is warmed and served in unopened pkg. but is the Patty prepared on a dedicated or cleaned grill area?
1200 Dekalb Ave, Sycamore, IL 60178
The have a "Nutrition & Allergen Guide" including a gluten free section by the cash registers.
5742 Tylersville Rd, Mason, OH 45040
Great burgers on gluten free buns! Also GF sides available and frozen custard . Very clean and family friendly .
4036 SW 30th Pl, Gainesville, FL 32608
A warm Gluten free bun is individually packaged and provided separately to be prepared to avoid any cross-contamination. Staff takes the extra time to let you know what sides are safe. Best place to go for a burger and shake!
2573 Commercial Blvd, Chippewa Falls, WI 54729
My celiac family loves eating here. They truly understand cross contamination and love their burgers. It's nice they have options that are GF for sides but I wish they would get a dedicated gf fryer for French fries. Love their ice cream options too.
1431 Johns Lake Rd, Clermont, FL 34711
Much like almost all Culver's Restaurants in the midwest, this one is very knowledgeable about Celiac's and cross-contamination. They do a great job of noting a food allergy, and the burger comes with a gluten-free bun that my son (who has CD) says is very good. You can also get mashed potatoes and other sides. All in all Culver's is always one of our saving graces when trying to find a safe place to eat!
1815 East Primrose Street, Springfield, MO 65804
My husband stopped on the way home and I didn't have time to look anything up. He asked about gf and they have gf buns! My burger came with the toppings and meat in the box and a bun still sealed in the plastic package!
1700 Bus Hwy 18-151 E, Mt. Horeb, WI 53572
I do not have celiac, but am gluten-intolerant. I went through the drive-thru, asked for a gluten-free bun with my burger. The bacon burger came separate in a container with all the condiments on the side. The gluten-free bun was still wrapped in the package with an easily identifiable label. It was good and I was comfortable.
9790 Washington Street, Thornton, CO 80229
Trendy GF ONLY! For anyone with an actual allergy or celiac this place is a huge no go.
309 SW Pine Island Rd, Cape Coral, FL 33993
Not all Culver's have a dedicated fryer. This one does! Ask for their allergy booklet to find several choices. It's nice to have a fast food place that truly offers GF. Also, you can get GF buns.
4140 Rusty Road, St Louis, MO 63129
Gluten free bun, knowledgeable about cross contamination and I've never gotten sick from there! Love!
10301 Corkscrew Rd, Estero, FL 33928
I love their double butter burger on a gluten free roll. The roll is $1.50 extra but it is delicious. Their hamburgs are wonderful too. If you are old you get 10 percent off for a senior discount if you ask for it.
Another reviewer mentioned a dedicated fryer but last time I asked it was a shared fryer -- I suggest you ask. It was the owner that helped me last time so I don't think I misunderstood (and I told him Chick-fil-a has a dedicated fryer to pressure him to get one). The bun is an Udi bun. They bring the burger to you and you have to assemble it yourself as the bun is still in the plastic bag. I find the gluten free buns change the flavor profiles of burgers so what tastes good with a wheat bun does not really work as a GF burger. I also find you need more toppings to get more flavor.
1540 N Verrado Way, Buckeye, AZ 85396
They serve your hamburger or what ever you order with the bun separately. You have to put your own condiments on. But you have choices on burgers or sandwiches. Plus the soups are gluten-free. Not the fries, the fries are contaminated!! But you can coleslaw if you don't want soup. It's pricey for your meal for its nice once in a while plus you can have their ice cream.
3400 W Kimberly Rd, Davenport, IA 52806
I've been to the Davenport location on multiple occasions. Not only do I get a gf bun steamed in it's closed bag, but they brought out my food on a separate tray.
1040 W Wade Hampton Blvd, Greer, SC 29651
The burgers are great, they have gluten free buns available and they made sure to ask if it was an allergy. Manager told me the fries were used in the same fryer. Manger was very helpful and food was good.
4121 Pioneer Woods Dr, Lincoln, NE 68506
My husband has celiac disease. He had a double deluxe on the gluten free bun, mashed potatoes without gravy, and cole slaw with no problem. I asked about the dedicated fryer for fries and the employee told me that they occasionally put chicken in there, so I didn't get fries. Not sure if he knew what he was talking about or not so I didn't risk it.
90 Cattlemen Rd, Sarasota, FL 34232
Great staff. Pre-wrapped GF buns, allergen menus at the counter, options for sandwiches, sides, and custard. My Reuben was excellent and I didn't get sick. The manager said that the fries are fried separately but something about how they handle the oil can result in a chance of cross-contamination. It's so exciting to have a fast place for good, safe food. My non-GF husband loved his burger and fries.
2121 W Main St, Rapid City, SD 57702
One of my favorite places to eat simply for the gluten free buns & custard!
3604 University Dr, Muscatine, IA 52761
If you are celiac, you can get a burger with gluten-free bun. If it's more of a preference, you can also have the fries, but there may be some cc.
Great to have a fast food option for those who are very sensitive. They have a GF menu, staff was very knowledgeable, and the GF bun was great!
124 Grant Way, New Richmond, WI 54017
they offer gluten free buns and they can flag your food as an allergy. it is not guaranteed that it would be 100% safe, but it's worth a shot. everything in the fryer can be cross contaminated.
9790 Washington Street, Thornton, CO 80229
The Cashew chicken salad without croutons, was very delicious.
2101 Lake Shore Dr E, Ashland, WI 54806
Would be happier if they had a dedicated fryer for French fries.
2302 S. Neil Street, Champaign, IL 61820
Amazing gluten free buns! My favorite is the double bacon cheeseburger on a gluten free bun.
3115 Towne Centre Boulevard, Lansing, MI 48912
Love the giant GF buns! My go-to fast food burger spot!
371 Airport Rd, Arden, NC 28704
Excellent gluten free buns, knowledgeable about celiac, gluten free menu linked on this app. Willing to scrub grill for grilled chicken.
371 Airport Rd, Arden, NC 28704
They are great! GF knowledgeable and cleaned grill for our chicken. Their GF buns are really good and just $1.19 extra for the bun.
Best burger I have had in a long time! Nice, heated soft bun that is packaged separately to avoid cross contamination! Dedicated fryer if you want fries. Nice looking salads I'm going to try soon!
5901 NW 64th St, Kansas City, MO 64151
Their web site seems to say their fries are gluten free, but the store staff, one of the cooks, said they were fried in the same fryer as the onion rings. We left.
2750 Muegge Rd, St Charles, MO 63303
I used to work here long ago so I know they take gluten allergies very seriously (all allergies!) I was ecstatic to find the gluten free bun and everything packaged separately. My only qualm is the GF bun is a little dry but their sauces fix that. I'm happy to have such a GF friendly fast food place so close!
1239 Dell Range Blvd, Cheyenne, WY 82009
From their website (9/24/16): Culver's restaurants contain common allergens and our dishes are prepared to order; however, normal kitchen operations involve shared cooking and preparation areas and we cannot guarantee any menu item will be free of allergens. Items cooked in the fryers may come into contact with egg, milk, wheat/gluten, soy, fish and shrimp. Culver's ground beef is 100% beef, and it is cooked on a grill with bread.
7841 S Howell Ave, Oak Creek, WI 53154
Gluten Free buns that are wrapped, ingredients and burger patty are packed separately and do not interact with any gluten.
7841 S Howell Ave, Oak Creek, WI 53154
Gluten free buns can be substituted for wheat bun.
1312 W McGalliard Rd, Muncie, IN 47303
They offer Udi's buns. They are prepackaged and then you get your sandwich fillers in a different box. You then get to put the sandwich together.
5520 N Main St, Mishawaka, IN 46545
They have warmed Udi's gluten free buns that they serve in a separate bag. The burgers and toppings were served separately. Dedicated fryer for the fries. My daughter has celiac disease and she had her first fast food burger in over 3 years. She loves their custard too. The staff was very knowledgeable about cross contamination.
1800 E Summit St, Crown Point, IN 46307
Culvers takes care of me every time! I love the deluxe burger on a GF bun with a side salad. They even warm the bun!!! They are my go to place for GF burger on the run and best of all I have NEVER gotten glutened/sick from eating here!
Monday | 10:00 am - 10:00 pm |
Tuesday | 10:00 am - 10:00 pm |
Wednesday | 10:00 am - 10:00 pm |
Thursday | 10:00 am - 10:00 pm |
Friday | 10:00 am - 10:00 pm |
Saturday | 10:00 am - 10:00 pm |
Sunday | 10:00 am - 10:00 pm |
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