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PyroTRF-ID: a novel bioinformatics methodology for the affiliation of terminal-restriction fragments using 16S rRNA gene pyrosequencing data
Ecological and evolutionary microbiology
In molecular microbial ecology, massive sequencing is gradually replacing classical fingerprinting techniques such as terminal-restriction fragment length polymorphism (T-RFLP) combined with cloning-sequencing for the characterization of microbiomes. Here, a bioinformatics methodology for pyrosequencing-based T-RF identification (PyroTRF-ID) was developed to combine pyrosequencing and T-RFLP approaches for the description of microbial communities. The strength of this methodology relies on the identification of T-RFs by comparison of experimental and digital T-RFLP profiles obtained from the same samples. DNA extracts were subjected to amplification of the 16S rRNA gene pool, T-RFLP with the
PyroTRF-ID was efficient for high-throughput mapping and digital T-RFLP profiling of pyrosequencing datasets. After denoising, a dataset comprising ca. 10′000 reads of 300 to 500 bp was typically processed within ca. 20 minutes on a high-performance computing cluster, running on a Linux-related CentOS 5.5 operating system, enabling parallel processing of multiple samples. Both digital and experimental T-RFLP profiles were aligned with maximum cross-correlation coefficients of 0.71 and 0.92 for high- and low-complexity environments, respectively. On average, 63±18% of all experimental T-RFs (30 to 93 peaks per sample) were affiliated to phylotypes.
PyroTRF-ID profits from complementary advantages of pyrosequencing and T-RFLP and is particularly adapted for optimizing laboratory and computational efforts to describe microbial communities and their dynamics in any biological system. The high resolution of the microbial community composition is provided by pyrosequencing, which can be performed on a restricted set of selected samples, whereas T-RFLP enables simultaneous fingerprinting of numerous samples at relatively low cost and is especially adapted for routine analysis and follow-up of microbial communities on the long run.
Molecular microbial ecology has become an important discipline in natural and medical sciences. Research on the structure, dynamics and evolution of microbial communities in environmental, human, and engineered systems provides substantial scientific knowledge for understanding the underlying microbial processes, for predicting their behavior, and for controlling, favoring, or suppressing target populations
Different analytical methods have been successively developed for the assessment of microbial communities via profiling or metagenomic approaches
If T-RFLP and NGS are meant to be used concomitantly for the investigation of microbial systems, one key objective is to link T-RFs to phylotypes. In parallel to early developments of T-RFLP methods, several computational procedures have been proposed to predict T-RF sizes and to phylogenetically affiliate T-RFs. For instance, TAP T-RFLP
In the current study, we propose a novel bioinformatics methodology, called PyroTRF-ID, to assign phylogenetic affiliations to experimental T-RFs by coupling pyrosequencing and T-RFLP datasets obtained from the same biological samples. A recent study showing that natural bacterial community structures analyzed with both techniques were very similar
Two different biological systems were used for analytical procedure validation. The first set comprised ten groundwater (GRW) samples from two different chloroethene-contaminated aquifers that have been previously described by Aeppli et al.
DNA extraction
GRW samples were filtered through 0.2-μm autoclaved polycarbonate membranes (Isopore™ Membrane Filters, Millipore) with a mobile filtration system (Filter Funnel Manifolds, Pall Corporation). DNA was extracted using the PowerSoil™ DNA Extraction Kit (Mo-Bio Laboratories, Inc.) following the manufacturer instructions, except that the samples were processed in a bead-beater (Fastprep FP120, Bio101) at 4.5 m·s-1 for 30 s after the addition of solution C1.
DNA from AGS samples was extracted with the automated Maxwell 16 Tissue DNA Purification System (Promega, Duebendorf, Switzerland) according to manufacturer′s instructions with following modifications. An aliquot of 100 mg of ground granular sludge was preliminarily digested during 1 h at 37°C in 500 μL of a solution composed of 5 mg·mL-1 lysozyme in TE buffer (10 mM Tris–HCl, 0.1 mM EDTA, pH 7.5). The DNA extracts were resuspended in 300 μL of TE buffer.
All extracted DNA samples were quantified with the ND-1000 Nanodrop® spectrophotometer (Thermo Fisher Scientific, USA) and stored at −20°C until analysis.
Experimental T-RFLP
The eT-RFLP analysis of the GRW series was done according to Rossi et al.
The resulting eT-RFLP profiles were generated between 50 and 500 bp as described in
Cloning and sequencing
Clone libraries were constructed with the 16S rRNA gene pool amplified from DNA samples using the same PCR procedures as described in the eT-RFLP method but with an unlabeled 8f primer. The PCR products were purified with the purification kit Montage® PCR Centrifugal Filter Devices (Millipore, USA), ligated into pGEM®-T Easy vector (Promega, USA) and transformed into
A total of 15 biological samples were analyzed using bacterial tag encoded FLX amplicon pyrosequencing analysis. A first set of DNA extracts from GRW and AGS samples were sent for sequencing to Research and Testing Laboratory LLC (Lubbock, TX, USA). The samples underwent partial amplification of the V1-V3 region of the 16S rRNA gene by PCR with unlabeled 8f and 518r primers, secondary PCR with tagged fusion primers for FLX amplicon sequencing, emulsion-based clonal amplification (emPCR), and GS FLX sequencing targeting at least 3′000 reads with the 454 GS-FLX Titanium Genome Sequencing System technology (Roche, Switzerland). The whole sample preparation protocol has been made available by the company in the publication of Sun et al.
A second set of amplicons from different GRW samples was analyzed by GATC Biotech AG (Konstanz, Germany) following an analog procedure but targeting at least 10′000 reads (referred to as the high reads amount method, HighRA, hereafter). The A- and B-adapters for sequencing with the Roche technology were ligated to the ends of the DNA fragments. The samples were run on a 2% agarose gel with TAE buffer and the band in a size range of 700–900 bp, 450–650 bp, or 100–500 bp, respectively, was excised and column purified. After concentration measurement the differently tagged libraries were pooled. The three resulting library pools were immobilized onto DNA capture beads and the amplicon-beads obtained were amplified through emPCR according to the manufacturer′s recommendations. Following amplification, the emulsion was chemically broken and the beads carrying the amplified DNA library were recovered and washed by filtration. Each pool was sequenced on a quarter GS FLX Pico-Titer plate device with GS FLX Titanium XLR70 chemistry on a GS FLX+ Instrument. The GS FLX System Software Version 2.6 was used and the GS FLX produced the sequence data as Standard Flowgram Format (SFF) file containing flowgrams for each read with basecalls and per-base quality scores.
Development of the PyroTRF-ID bioinformatics methodology
The PyroTRF-ID bioinformatics methodology for identification of T-RFs from pyrosequencing datasets was coded in Python for compatibility with the
Figure 1
Data workflow in the PyroTRF-ID bioinformatics methodology
Data workflow in the PyroTRF-ID bioinformatics methodology. Experimental pyrosequencing and T-RFLP input datasets (
Input files
Input 454 tag-encoded pyrosequencing datasets were used either in raw standard flowgram (.sff), or as pre-denoised fasta format (.fasta) as presented below. Input eT-RFLP datasets were provided in coma-separated-values format (.csv).
Sequence denoising was integrated in the PyroTRF-ID workflow but this feature can be disabled by the user. It requires the independent installation of the QIIME software
Mapping of sequences was performed using the Burrows-Wheeler Aligner′s Smith-Waterman (BWA-SW) alignment algorithm
Generation of dT-RFLP profiles
Sequences that passed through all previous steps of the procedure were digested
Comparison of eT-RFLP and dT-RFLP profiles
In order to allow comparison with eT-RFLP profiles, T-RFs below 50 bp were removed, and a second set of dT-RFLP profiles was generated. To overcome any possible discrepancy between experimental and
Assignment of affiliation to dT-RFs
Peak annotation files were generated in comma-separated-values format (.csv), listing all digitally obtained T-RFs within each dT-RFLP profile, together with their original and shifted lengths. Closest phylogenetic affiliations were provided together with the number of reads and their relative contribution to the T-RF, as well as with the absolute and normalized SW mapping scores, and the Genbank code of each reference sequence. When eT-RFLP data were not provided in the workflow, the peak annotation file was directly obtained after dT-RFLP processing without removing dT-RFs below 50 bp and without indication of T-RF shift.
Optimization and testing of PyroTRF-ID
The initial testing and validation steps were carried out with the 17 pyrosequencing datasets originating from the two environments. The impact of the data processing steps of the PyroTRF-ID pipeline was assessed using two samples (GRW01 and AGS01). Three different combinations of algorithms were tested for the processing of sequences (Table
The optimal procedure was then applied for the comparison of PyroTRF-ID results obtained from groundwater and wastewater environments. Finally, restriction enzymes commonly used in T-RFLP analyses of bacterial communities (
Pyrosequencing quality control and read length limitation
The principal quality outputs given by PyroTRF-ID are presented in Additional file
Additional file 1
Quality plots generated for samples pyrosequenced with LowRA (>3′000 reads) and HighRA methods (>10′000 reads). Sequence quality PHRED scores over all bases (A): PHRED scores are defined as the logarithm of the base-calling error probability Perror = 10-PHRED/10 and PHRED = −10 log Perror. Box plots represent the distribution of reads quality at each sequence length. The black curve represents the mean sequence quality in function of the sequence length. Distribution of the mean sequence quality PHRED score over the pyrosequencing reads (B). Distribution of sequence lengths over all pyrosequencing reads (C). Only sequences between 300 and 500 bp were kept for dT-RFLP analysis.
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Filtering based on the PHRED quality criterion allowed discarding low quality sequences. Most of the remaining sequences had a length below 400–450 bp (Additional file
Effect of denoising and mapping procedures
Denoising of pyrosequencing datasets was performed in order to correct for classical 454 analytical errors including the above-mentioned cut-off values: a minimum PHRED quality score of 20, as well as minimum and maximum sequence lengths of 300 and 500 bp, respectively. The denoising process generated a subset of representative sequences harboring at least 3% dissimilarity to each other. This amounted to 17±1% and 43±9% of the number of reads present in the raw datasets obtained with the HighRA and LowRA methods, respectively.
After denoising, the mapping process was the time-limiting step in the PyroTRF-ID pipeline. Twenty minutes were required for mapping the largest datasets against the Greengenes database. Discarding sequences shorter than 300 bp did not lead to a reduced number of detected bacterial phylotypes (Additional file
Additional file 2
Assessment of mapping performances with pyrosequencing datasets denoised without (0–500 bp) and with (300–500 bp) minimal read length cutoff. Examples are given for the groundwater sample GRW01, the flocculent activated sludge sample FLS01 and the aerobic granular sludge sample AGS01. After denoising with the one or the other method, each dataset was mapped against a reference database with MG-RAST
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After PHRED-filtering, the remaining raw sequences had maximum lengths of 450 bp and therefore the maximal SW mapping scores amounted to around 450. The distributions of the absolute and normalized SW scores are provided in Additional file
Additional file 3
Comparison of the distributions of the SW mapping score and of the traditional identity score used by microbial ecologists in the field of environmental sciences for phylogenetic affiliation of sequences. The distributions of the absolute SW score (A) and of the SW scores normalized by the read lengths (B) obtained after mapping of 15 pyrosequencing datasets with the BWA-SW algorithm implemented in the PyroTRF-ID methodology are compared to the distribution of the identity score obtained after annotation of 10 pyrosequencing datasets with MG-RAST
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Generation of digital T-RFLP profiles
The dT-RFLP profiles were successfully generated with the standard PyroTRF-ID procedure (Table
Additional file 4
Full digital T-RFLP profiles. Examples of full digital T-RFLP profiles obtained with the restriction enzymes
Click here for file
An additional feature of PyroTRF-ID is the generation of dT-RFLP profiles with any restriction enzyme. Here profiles were obtained with five additional restriction enzymes and compared. Profiles of GRW samples were on average 2.3 times richer than ones of AGS samples, and each restriction enzyme generated characteristic dT-RFLP features regardless of the sample complexity (Figure
Figure 2
Density plots displaying the repartition of T-RFs along the 0–500 bp domain with different endonucleases
Density plots displaying the repartition of T-RFs along the 0–500 bp domain with different endonucleases. The effect of the different restriction endonucleases
Comparison of digital and experimental T-RFLP profiles
Mirror plots generated by PyroTRF-ID computed with raw and denoised pyrosequencing datasets obtained from a complex bacterial community (GRW01) are presented in Figure
Additional file 5
Comparison of mirror plots obtained on raw (left) and on denoised (right) pyrosequencing datasets. Examples are given for the sample GRW01 pyrosequenced with the HighRA method (A) and for the samples GRW07 (B) and AGS01 (C) pyrosequenced with the LowRA method.
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Figure 3
Mirror plot displaying the cross-correlation between digital and experimental T-RFLP profiles
Mirror plot displaying the cross-correlation between digital and experimental T-RFLP profiles. This mirror plot was generated for the complex bacterial community of sample GRW01. Comparison of mirror plots constructed with raw (A) and denoised sequences (B). Relative abundances are displayed up to 5% absolute values. For those T-RFs exceeding these limits, the actual relative abundance is displayed beside the peak.
The dT-RFLP profiles exhibited a drift of 4 to 6 bp compared to eT-RFLP profiles. From
Additional file 6
Assessment of cross-correlation and optimal lag between denoised dT-RFLP and eT-RFLP profiles. The denoised dT-RFLP profiles of the samples AGS07 (A) and GRW04 (B) were both shifted with optimal lags of −5 bp to match with the related eT-RFLP profiles. At these optimal lags, the maximum cross-correlation coefficients amounted to 0.91 (AGS07) and 0.71 (GRW04).
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Impact of sequence processing steps, pyrosequencing methods and sample types
Indices of richness (number of T-RFs) and diversity (number of T-RFs and distributions of abundances) were used to evaluate the impacts of data processing steps, pyrosequencing methods and sample types on the structure of the final dT-RFLP profiles (Figure
Figure 4
Assessment of the impact of data processing on dT-RFLP profiles, and comparison with eT-RFLP profiles
Assessment of the impact of data processing on dT-RFLP profiles, and comparison with eT-RFLP profiles. Richness and Shannon′s H′ diversity indices were calculated in a way to quantify the impact of the pyrosequencing data processing parameters on the resulting dT-RFLP profiles. Two examples are given for samples pyrosequenced with the HighRA (GRW01) and LowRA methods (GRW07).
The DNA extract of one AGS sample was analyzed in triplicate from pyrosequencing to PyroTRF-ID. The resulting standard dT-RFLP profiles contained 94±10 T-RFs, and exhibited very close diversity indices of 1.48±0.03. In comparison, denoised profiles of all AGS samples collected over 50 days contained similar numbers of T-RFs (84±9) but exhibited quite different diversity indices of 2.12±0.48. There was also very little variation in the cross-correlation coefficients (0.90±0.01) between the dT-RFLP profiles and the corresponding eT-RFLP profile. All three denoised T-RFLP profiles exhibited similar structures, and affiliations were the same for T-RFs that could be identified.
Efficiency of phylogenetic affiliation of T-RFs
Comprehensive phylogenetic information was provided by PyroTRF-ID for each dT-RF, as exemplified in Table
Figure 5
Amount of bacterial affiliations contributing to T-RFs
Amount of bacterial affiliations contributing to T-RFs. The absolute (A) and relative numbers (B) of T-RFs that comprised one to several bacterial affiliations is given for the samples GRW01 and AGS01.
Some reference sequences were sometimes represented by several T-RFs (Table
Additional file 7
Alignment of sequences mapping with the same reference sequence with identical accession number in the Greengenes database, and resulting in different digital T-RFs. Examples are given for the
Click here for file
Advantages and novelties of the PyroTRF-ID bioinformatics methodology
This study describes the development of the PyroTRF-ID bioinformatics methodology for the analysis of microbial community structures, and its application on low- and high-complexity environments. PyroTRF-ID can be seen as the core of a high-throughput methodology for assessing microbial community structures and their dynamics combining NGS technologies and more traditional community fingerprinting techniques such as T-RFLP. More than just predicting the most probable T-RF size of target phylotypes, PyroTRF-ID allows the generation of dT-RFLP profiles from 16S rRNA gene pyrosequencing datasets and the identification of experimental T-RFs by comparing dT-RFLP to eT-RFLP profiles constructed from the same DNA samples.
At the initial stage of the assessment of a microbial community, PyroTRF-ID can be used for the design of an eT-RFLP procedure adapted to a given microbial community through digital screening of restriction enzymes. In contrast to previous studies involving
Subsequently, eT-RFLP can be used as a routine method to assess the dynamics of the stuctures of microbial communites, avoiding the need for systematic pyrosequencing analyses. We suggest that pyrosequencing should be applied at selected time intervals or on representative samples to ensure that the T-RFs still display the same phylogenetic composition. Combining T-RFLP and pyrosequencing is particularly adapted for the temporal follow-up of a microbial system, taking advantage of the relative low costs of T-RFLP and its convenience for routine assessment of microbial community structures, and of the power of pyrosequencing for assessing the composition of these communities. PyroTRF-ID has already been used for the study of bacterial communities involved in start-up of aerobic granular sludge systems
Performance assessment and limitations of PyroTRF-ID
Classical 454 pyrosequencing errors, such as, inaccurate resolving of homopolymers and single base insertions
Clipping the low-quality end parts of sequences is an option to improve sequence quality but it is quite improbable that it has an impact on the outcome of the taxon assignment and the creation of dT-RFLP profile. When PyroTRF-ID is run with the "--qiime" option, quality trimming is done using the protocol proposed in QIIME
Discrepancies of 0–7 bp between the size of
Following the standard PyroTRF-ID procedure, high level of correspondence was obtained between dT-RFLP and eT-RFLP profiles. Over all samples, 63±18% of all eT-RFs could be affiliated with a corresponding dT-RF. Correspondence between dT-RFs and eT-RFs was relatively obvious for high abundance T-RFs, in contrast to low abundance dT-RFs. Numerous low abundance dT-RFs were present in dT-RFLP profiles but absent in eT-RFLP profiles. Conversely, eT-RFs were sometimes lacking a corresponding dT-RF. This mainly occurred in profiles generated using pyrosequencing datasets with an initially low amount of reads exceeding 400 bp. The lower proportion of long reads was associated with a decreasing probability of finding a restriction site in the final portion of the sequences. For eT-RFs near 500 bp, incomplete enzymatic restriction could explain that undigested amplicons were detected in the electrophoresis runs
The majority of dT-RFs were affiliated to several phylotypes, revealing the underlying phylogenetic complexity, which was in agreement with Kitts
This study presented the successful development of the PyroTRF-ID bioinformatics methodology for high-throughput generation of digital T-RFLP profiles from massive sequencing datasets and for assigning phylotypes to eT-RFs based on pyrosequences obtained from the same samples. In addition, this study leads to the following conclusions:
• The combination of pyrosequencing and eT-RFLP data directly obtained from the same samples was a powerful characteristic of the PyroTRF-ID methodology, enabling generation of dT-RFLP profiles that integrate the whole complexity of microbiomes of interest.
• The LowRA and HighRA 454 pyrosequencing method did not impact on the final results of the PyroTRF-ID procedure.
• As in any new generation sequencing analysis, denoising was a crucial step in the 454 pyrosequencing dataset processing pipeline in order to generate representative digital fingerprints.
• The PyroTRF-ID workflow could be applied to the screening of restriction enzymes for the optimization of favorably distributed eT-RFLP profiles by considering the entire underlying microbial communities.
• The PyroTRF-ID methodology was validated with different samples from low- and high-complexity environments, and could be implemented in a broad spectrum of biological samples in environmental to medical applications with optimized laboratory and computational costs. This methodology is probably not restricted to pyrosequencing datasets, and could be, after some modifications, applied to datasets obtained with any kind of sequencing techniques.
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
DGW and NS equally contributed to this work by conceiving, designing and coordinating the study, by carrying out sampling and molecular biology investigations, by leading the development of the PyroTRF-ID bioinformatics methodology, by analyzing all collected data, and by drafting the manuscript. DGW additionally conceived the tables and the figures. LS was responsible for the optimization and validation of PyroTRF-ID and wrote the underlying codes. GL coded the initial bioinformatics procedure. JM and PR participated in the design of the study. JR coordinated the development of PyroTRF-ID at the Bioinformatics and Biostatistics Core Facility. CH led the project and gave the initial idea of reconstructing T-RFLP profiles from pyrosequencing data. DGW and NS wrote the manuscript, with additional contributions of JM, PR, and CH. All authors read and approved the final manuscript.
This research was financed by the Swiss National Science Foundation, Grants No. 120536, 138148 and 120627. We recognize the excellent assistance of Yoan Rappaz in molecular biology analyses. We acknowledge Scot E. Dowd, Yan Sun, Lars Koenig and at Research and Testing Laboratory (Lubbock, Texas, USA), Timothy M. Vogel, Sébastien Cecillon and the Environmental Microbial Genomics Group at Ecole Centrale de Lyon (France), and GATC Biotech (Konstanz, Germany) for pyrosequencing analyses and advice. We are grateful to Ioannis Xenarios for support and access to the Vital-IT HPCC of the Swiss Institute of Bioinformatics (Lausanne, Switzerland).
Assessment and management of soil microbial community structure for disease suppression
Synchrony in aquatic microbial community dynamics
Molecular methods in biological systems
Advances in the use of terminal restriction fragment length polymorphism (T-RFLP) analysis of 16S rRNA genes to characterize microbial communities
Terminal restriction fragment length polymorphism (T-RFLP): an emerging method for characterizing diversity among homologous populations of amplification products
Adaption of a fragment analysis technique to an automated high-throughput multicapillary electrophoresis device for the precise qualitative and quantitative characterization of microbial communities
Soil microbial community analysis using terminal restriction fragment length polymorphisms
Statistical assessment of variability of terminal restriction fragment length polymorphism analysis applied to complex microbial communities
Cloning method for taxonomic interpretation of T-RFLP patterns
Name that microbe: rapid identification of taxa responsible for individual fragments in fingerprints of microbial community structure
Changes in N-transforming archaea and bacteria in soil during the establishment of bioenergy crops
Pyrosequencing sheds light on DNA sequencing
Tag-encoded FLX amplicon pyrosequencing for the elucidation of microbial and functional gene diversity in any environment
Metagenomic pyrosequencing and microbial identification
Pyrosequencing enumerates and contrasts soil microbial diversity
Metagenomics: read length matters
Testing the limits of 454 pyrotag sequencing: reproducibility, quantitative assessment and comparison to T-RFLP fingerprinting of aquifer microbes
Field guide to next-generation DNA sequencers
Data handling strategies for high throughput pyrosequencers
A Bioinformatician′s guide to metagenomics
The smallest cells pose the biggest problems: high-performance computing and the analysis of metagenome sequence data
From genomics to metagenomics
Validating T-RFLP as a sensitive and high-throughput approach to assess bacterial diversity patterns in human anterior nares
REPK: an analytical web server to select restriction endonucleases for terminal restriction fragment length polymorphism analysis
Use of pyrosequencing and denaturing gradient gel electrophoresis to examine the effects of probiotics and essential oil blends on digestive microflora in broilers under mixed eimeria infection
Short-term antibiotic treatment has differing long-term impacts on the human throat and gut microbiome
Bacterial diversity across individual lichens
Terminal restriction fragment length polymorphism analysis program, a web-based research tool for microbial community analysis
TRiFLe, a program for in silico terminal restriction fragment length polymorphism analysis with user-defined sequence sets
T-RFPred: a nucleotide sequence size prediction tool for microbial community description based on terminal-restriction fragment length polymorphism chromatograms
Quantifying in situ transformation rates of chlorinated ethenes by combining compound-specific stable isotope analysis, groundwater dating, and carbon isotope mass balances
PhD thesis #5379
Bacterial selection during the formation of early-stage aerobic granules in wastewater treatment systems operated under wash-out dynamics
Performance and microbial community composition dynamics of aerobic granular sludge from sequencing batch bubble column reactors operated at 20°C, 30°C, and 35°C
Ordination and significance testing of microbial community composition derived from terminal restriction fragment length polymorphisms: application of multivariate statistics
Development of degenerate and specific PCR primers for the detection and isolation of known and putative chloroethene reductive dehalogenase genes
BioEdit: a user-friendly biological sequence alignment editor and analysis program for windows 95/98/NT
Bellerophon: a program to detect chimeric sequences in multiple sequence alignments
Open software for biologists: from famine to feast
QIIME allows analysis of high-throughput community sequencing data
Base-calling of automated sequencer traces using phred. II. Error probabilities
Systematic exploration of error sources in pyrosequencing flowgram data
Accurate determination of microbial diversity from 454 pyrosequencing data
Rapidly denoising pyrosequencing amplicon reads by exploiting rank-abundance distributions
Fast and accurate long-read alignment with burrows-wheeler transform
An improved greengenes taxonomy with explicit ranks for ecological and evolutionary analyses of bacteria and archaea
Identification of common molecular subsequences
Assessing annotation transfer for genomics: quantifying the relations between protein sequence, structure and function through traditional and probabilistic scores
Geobiological analysis using whole genome-based tree building applied to the bacteria, archaea, and eukarya
Accuracy and quality of massively parallel DNA pyrosequencing
Wrinkles in the rare biosphere: pyrosequencing errors can lead to artificial inflation of diversity estimates
Artificial and natural duplicates in pyrosequencing reads of metagenomic data
Recharacterization of ancient DNA miscoding lesions: insights in the era of sequencing-by-synthesis
Removing noise from pyrosequenced amplicons
Terminal restriction fragment patterns: a tool for comparing microbial communities and assessing community dynamics
Terminal restriction fragment length measurement errors are affected mainly by fragment length, G plus C nucleotide content and secondary structure melting point
Variation between observed and true terminal restriction fragment length is dependent on true TRF length and purine content
An evaluation of terminal-restriction fragment length polymorphism (T-RFLP) analysis for the study of microbial community structure and dynamics
Terminal restriction fragment patterns (TRFPs), a rapid, PCR-based method for the comparison of complex bacterial communities
Formation of pseudo-terminal restriction fragments, a PCR-related bias affecting terminal restriction fragment length polymorphism analysis of microbial community structure
Electron acceptor-dependent identification of key anaerobic toluene degraders at a tar-oil-contaminated aquifer by pyro-SIP
The metagenomics RAST server - a public resource for the automatic phylogenetic and functional analysis of metagenomes
The ribosomal database project: improved alignments and new tools for rRNA analysis
Identifying biologically relevant differences between metagenomic communities


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Author Archive
CENLA Information
by BruceF on Nov.08, 2015, under Events
Live Scoring Click HERE
Tonight's the Night CENLA Autocross
Yes…CENLA is going to have a NIGHT autocross at Lamar Dixon. It will be our last event for this "season". Our next event will be September 17, 2017.
This will be the first NIGHT event in Louisiana in about 15-20 years! (we don't think anyone remembers for sure). So when will the next one be? Actually we will have another on October 14, 2017. But DON'T miss the FIRST one!
Karts ARE welcome at this location!
Cash or checks or Credit Cards.…
$30 SCCA Members
$40 Non SCCA Members
$10 Penalty for registration on-site and a stern look from timing and scoring
Autocross Schedule:
-2:00 PM: Setup
-5:30 PM-7:00 PM: Registration and Tech
-7:00 PM Novice walk-thru
-7:30 PM: Driver's Meeting
-8:00 PM: First Car Out
Sundown 7:51 PM
2017 event schedule
CENLA does not run June-August but we will have other things happening!
September 17 (Sunday)
October 14 NIGHT EVENT! (Saturday)
November 5 (Sunday)
December 3 (Sunday)
NEW for this year!
"Yearly Tech/Wavier"
For a fee of $40, you can get sign the new yearly wavier form and get your car teched once! There will also be a special line (shorter) at the registration table just for these drivers.
We will still be doing a smaller spot check for those that are yearly teched but it will be VERY short.
So "how do you get in on this special deal?"
We will be starting this at the January 8th event. You MUST be at the registration table BY 8:30am. Latecomers will have to wait until the next event to sign up for this.
You may sign up for this at any event but will still pay the yearly fee of $40. So sign up early at the first event of the year!
Remember…all monies go back into the club fund to buy new equipment etc.
Any questions just post here. Answer to the first question that will be asked…yes…this has been ok'ed by SCCA.
Spring Tour Points Challenge
CENLA-SCCA will be holding "points events". Each regular event will be counted for points total. Prizes will be awarded at the end of each "Tour".…
1. January – May events (Spring Tour).
2. Driver must run at least 3 events to qualify for the points challenge.
3. Driver must be an SCCA member to qualify for the points challenge.
All drivers rankings will be posted.
4. Prizes will be announced at a later date.
5. There will also be a Fall Tour from September – December.
6. There will be awards for Class and PAX points.
7. Points will be awarded each event for classes as follows:
a. 1st place 10 points
b. 2nd place 7 points
c. 3rd place 5 points
d. 4th points 3 points
e. 5th place 1 point
So don't miss a single event!

subset: commoncrawl


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Phil Scott in victory lane last Thursday
Phil Scott in victory lane last Thursday
"We're still in this thing," said Scott. "We've been on a roll lately, and we're really building momentum." The Middlesex driver picked up his first victory of the season in last week's Vermont ACE Hardware Dealers feature and now trails leader Jamie Fisher by 57 points. "We had a tough early season, but we can chase the leaders down, it's been done before. The good part is that there's no pressure from where my team sits; the top three are fighting tooth-and-nail, and my job right now is to sneak into it."
Fisher, a Shelburne native, leads Dave Pembroke of Montpelier and Williamstown's Cris Michaud by a total spread of just seven points, the closest three-way battle in Thunder Road's 48-year history. Scott paces Becker by two points. In terms of sheer mathematics, fifteen drivers are still within the championship window with two races remaining. The final point-counting race for the Late Model division is at next Tuesday's Fisher/Federated Auto Parts event.


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Agarwal Girls T.T College - AGTTC Nagaur - AGTTC
Agarwal Girls T.T College.(B.Ed. College) is run by Jan Hit Sewa Sansthan,Near Old Post Office-Merta City registered under Rajasthan Society Act 1958 with the registrar of the societies Government of Rajasthan Vide Reg. No. Nag./Raj./184/2001-02 Dated 16-07-2002. and under consideration of N.C.T.E. This College is affiliated with the Maharshi dayanand Saraswati University,Ajmer , and recognitions with the N.C.T.E. for B.ED course intake of 100 students for the session 2008-09 under consideration The College is leading in imparting quality education in the field of teaching, various researches and community works. The whole system comprising of teachers, staff and students together try hard to achieve the target of the best possible knowledge and to transform themselves as per the changing environment of the society and to pursue intellectual and personal fulfillment. The modern teaching methodology assists the students in getting an all around view of engineering, technology and the entrepreneur world. The diligent faculty team of the college possesses hands-on experience and up to date knowledge of their subject and area of expertise. The infrastructure and facilities of Agarwal Girls T.T College consists of spread-out campus alongside various amenities in the form of library, computer center, classrooms with proper teaching and learning aids, medical facilities etc. The institute seeks to place each of its students in an organization/company of great repute.
Why join Agarwal Girls T.T College
The college has well state of the art infrastructure facilities.
1 Type Courses Available
- 1 UG Courses
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At Agarwal Girls T.T College (AGTTC) the students are endowed with all convenient amenities that make their life 'go easy' in college hours, alongside make learning more interesting. Some of the facilities include well ventilated classrooms, recreational areas, hostel facility for boys and girls, indoor and outdoor games, stocked library, medical facilities etc.
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Windlands South
3800 Sam Boney Dr, Nashville, TN 37211
Please call 1-800-741-0926 to speak to a Care Advisor
3800 Sam Boney Dr, Nashville, TN 37211
Please call 1-800-741-0926 to speak to a Care Advisor
Conveniently located just minutes from downtown Nashville, you'll love all the hospitality and friendliness Windlands South offers. Enjoy our warm summers, mild winters and all the colors of the seasons just outside your door. Explore all that Nashville has to offer with trips to the Loveless Café, the Parthenon, the Grand Old Opry and Bicentennial Park. We're conveniently close to shopping, several area hospitals and wonderful entertainment. Our live-in managers are here 24 hours a day to make you feel at home. You'll love coming home to us.
Windlands South is proud to be a Holiday Retirement community. Since 1971, Holiday Retirement has been creating superb residential communities where seniors feel a sense of home and belonging. This is what we like to call "The Holiday Touch."
Windlands South offers deluxe living in our studio, one- and two-bedroom apartment homes. Each apartment includes a private bathroom, patio and wall-to-wall carpeting. Our fully outfitted kitchens, complete with full-sized refrigerator, oven and dishwasher, allow you the option to dine in the privacy of your apartment or to join us in the community dining room for world-class cuisine prepared fresh by our culinary staff.
Complimentary weekly housekeeping and linen service, cable and utilities, laundry access and guest parking are included in your all-inclusive living experience.
While staying at our community, each resident has the opportunity to participate in our exclusive travel program. Through this program, you can visit nearly any of our over 300 retirement communities for up to seven nights at no additional charge. During your visit, you will enjoy all the same service and amenities you enjoy at home while having the chance to meet new friends and experience new places.
We understand that each individual is unique and that needs may change over time. Holiday does not provide any health care services. However, residents are welcome to receive services from any outside home health care provider of their choice to help them continue enjoying life at our community.
Windlands South offers everything you need for a rewarding and active retirement lifestyle.
3.67 out of 5.0
Family Or Friend Of A Current/Past Resident
Windlands South has a very nice lunchroom or restaurant. They seem to keep everything nice and clean. Every time I go to see Pat, her room looks good. They come up once a week to vacuum and dust. We are very happy with what they are offering. They have so many different choices of activities, including bingo, card games, and a movie theater. They had a dance before, and Pat likes to dance.
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Family Or Friend Of A Current/Past Resident
My father was in Windlands South until he passed away last month. He had a one-bedroom apartment with a full kitchen, and they had a shared dining room where he could have three meals a day, although the food was not very good. My father really did not participate, but they had activities for the residents and movie nights. The staff was accessible, but they were rude at times. They can improve by having better chefs in there.
3 people found this helpful. Was this helpful?
If you are looking for apartment style senior living, Windlands south is all that and more. Upon visiting, I was impressed by the caring and compassionate staff, an amazing chef created menu, a wide variety of activities including outings and close proximity to the plethora of exciting sites and sounds of Nashville. A beautiful facility that is truly unique in all that it has to offer.
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text:
Wednesday 9 September 1987
It was Wednesday, under the sign of Virgo. The US president was Ronald Reagan (Republican). In that special week of September people in US were listening to La Bamba by Los Lobos.
Fatal Attraction, directed by Adrian Lyne, was one of the most viewed movies released in 1987
while Presumed Innocent by Scott Turow was one of the best selling books.
But much more happened that day: find out below..
Or have a look at the whole 1987.
Old Newspapers
Have a look at the old newspapers from 9 September 1987 and get them!
Which were the most popular movies released in the last 30 days ?
TV Series
Which were the most popular TV series released this week ?
Find out your future
Get a FREE Numerology report based on the digits of 9 September 1987!
Wish him/her..
Historical Events
Which were the important events of 9 September 1987 ?
- North Korea - Republic Day (1948)
- Eastern Orthodoxy - Synaxis of the Theopatores Joachim and Anna
- California - Admission Day (to commemorate the state's admission to the USA)
- Japan - Chrysanthemum Day (Kiku no Sekku)
- Tajikistan - Independence Day (from USSR, 1991)
Famous Birthdays:
- Clayton Snyder: American actor.
- Joshua Herdman: English actor.
- Alexandre Song: Cameroonian footballer.
- Milan Stankovic: Serbian pop and folk singer.
- Timomatic : Pop Singer.
- Riley Cooper: Football Player
- Nick Van de Wall: DJ
- Adam Ali: YouTube Star
- Markus Jürgenson: Estonian footballer
- Alexis Palisson: French rugby player
- Andrea Petkovic: German tennis player
- Milan Stanković: Serbian singer
Famous Deaths:
- Gerrit John Heijn: Senior executive (Ahold), kidnapped / murdered.
- Gary Hart admits on Nightline, to cheating on his wife
- Largest US Tennis Open single night session (total) 21,016
- Larry Bird (Celtics), begins NBA free throw streak of 59
- Nolan Ryan strikes out his 4,500th batter.
Music Charts
Which were the top hits in that special week of September 1987 ?
Top #10 songs in the USA
- Los Lobos - La Bamba
- Michael Jackson - I Just Can't Stop Loving You
- Whitney Houston - Didn't We Almost Have It All
- Whitesnake - Here I Go Again
- Debbie Gibson - Only In My Dreams
- Dan Hill - Can't We Try
- Huey Lewis & The News - Doing It All For My Baby
- ABC - When Smokey Sings
- Madonna - Who's That Girl
- Bananarama - I Heard A Rumour
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...and if 9 September 1987 was your Birth Date then Join our Birthday Club!
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Magazine Covers
What news were making the headlines those days in September 1987?
- SportsIllustrated: Sports Illustrated: no.9080
- Time: TIME Magazine Cover: U.S. Catholics -- Sep. 7, 1987
- SportsIllustrated: Sports Illustrated: no.9079
Which were the most popular books released in the last weeks ?
Remember when
What videogames would you be playing back in the days ?


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Anonymous on Oct 18, 2012
I was one of 9 bridesmaids in a wedding, and the bride found these cute dresses. However, all the dresses (minus the one that never showed up!) were too large-thank goodness, though because the zipper didn't work and I had to pull it over my head. My dress only had lining on the top, and was see-through on the bottom. The pattern was beautiful, but the craftsmanship of this dress was shoddy. If we had got our dresses more than a week before the wedding, there might have been time to get them fixed. (for the most part) Anyway, we made it work with plenty of duct tape and safety pins, but it shouldn't have come to that.
Anonymous on Oct 16, 2012
Anonymous on Sep 8, 2012
Beautiful design and make
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ED Magedson – Founder
Ripoff Report
FirstData5565 Glenridge Connector NE, Atlanta, GA Internet United States of America
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This report was posted on Ripoff Report on 01/02/2013 12:45 PM and is a permanent record located here: http://www.ripoffreport.com/reports/firstdata/internet/firstdata-after-almost-3-years-they-want-to-charge-me-500-to-close-account-very-decepti-990030. The posting time indicated is Arizona local time. Arizona does not observe daylight savings so the post time may be Mountain or Pacific depending on the time of year.
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You are not alone
SUBMITTED: Saturday, April 18, 2015POSTED: Saturday, April 18, 2015
I'm a victim of FirstData's abuse and fraudulent activity. When I signed an agreement with CoastalPay (an ISO of FirstData) I refused to sign an equipment lease and stated on the agreement that I'm not leasing an equipment. They used my signature to forge a lease contract in my name, which they tried to use as a blackmail to force me to sign such contrat. I refused! I know I didn't sign the lease and stated it clearly in the agreement authorise them to process my credit cards. I know they can't not sue me because they risk to get their trick exposed. I'm not stopping there. I'm going to turn the table on them by sueing them myself for fraud, forging false documents and blackmail. Are you sure you filled and signed all the papers they claim you did? Did they give you a copy of all those documents upon signature with a rep? It's a well and sofisticated scam they are running and it's time to pull it off. I want you to join me.
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My Weekend was filled with food and gifts! The Best!
This weekend I decided to try something new for dinner. Paul and I really try to go to new places all the time. We have a few we just love and return too, but usually we're trying a new place. We tried Grand Szechuan this Friday, and it was very good. Saturday I hosted a "meet-up". If you guys haven't checked out the tone it up girls, they're great. Click here to see what their all about. There is a local group that is positive and motivating and occasionally have meetups. I hosted the latest one at Haute Barre on Saturday. Then Intern Katie and I tried out Yard House. It's the newest restaurant in the west end. We both really enjoyed it.
Sunday was my bridal shower here. I had such a lovely time. Paul's aunt can really host a fabulous party. Here are few pictures from my weekend!


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ED Magedson – Founder
Ripoff Report
National American UniversityOverland Park, KS Internet United States of America
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This report was posted on Ripoff Report on 11/04/2012 06:46 AM and is a permanent record located here: http://www.ripoffreport.com/reports/national-american-university/internet/national-american-university-are-degrees-worth-anything-op-internet-964473. The posting time indicated is Arizona local time. Arizona does not observe daylight savings so the post time may be Mountain or Pacific depending on the time of year.
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text:
Terribly Girly by MissV Photographer
Madame JenModel Female Atlanta, Georgia, US
My MM URL: http://www.modelmayhem.com/jenmcgrath
Mayhem # 237905
About Me
At the current time I work as the assistant and booking gal for Janette Valentine of www.terriblygirly.com. I'm a girl friday to my husband, Tattoo Artist, Timothy McGrath (www.bucktoothtimmy.com) and 13 Roses Tattoo Parlour.
I do this as a side project. I am heavily tattooed (with good work). So anything that relates thereto, I would be interested in. If it's not tattoo related then anything creative, especially vampy & dark which is right up my alley.... I enjoy being in front of the camera, as well as being behind the scenes of a shoot lending a hand. I've got a great personality, I'm reliable, good humored and enthusiastic. I come prepared with wardrobe and creative ideas.
I also make candles you can check that out at www.burningtearsbymadamejen.com
Be a fan on facebook: http://www.facebook.com/BuringTearsByMadameJen
Check out AGBF write up about me: http://agirlsbestfriend.biz/2011/07/12/ … adame-jen/
I'm also a big halloween enthusiast and make/hack halloween props, check out my blog: http://madamejen.blogspot.com/
If your interested in booking with Janette Valentine of Terribly Girly, please feel free to write me at email@example.com for more info!
Have you worked with Madame Jen? Add Credits for Madame Jen! >>
Credit Notes
www.varla.com *Varla Model*
Wicked Paper Dolls
Pinups For Troops
Antikulture Magazine Volume I
Tattoos for Women #75
Prick Magazine Sept 07 Ad for 13 Roses Tattoo Parlour
Prick Magazine Dec 07 Ad for 13 Roses Tattoo Parlour
Flagpole Magazine Dec 08 for 13 Roses Tattoo Parlour
Prick Magazine Jan 09 for 13 Roses Tattoo Parlour
Tattoo Flash July 2009
Skin & Ink Magazine April 2010
Pretty In Ink Magazine Issue #2
Rebelicious Magazine Issue #4
Tim McGrath
Kimberly Reed
Lindsey Rukkus
Terribly Girly by MissV Photographer
Kabuki Kosmetics Makeup Artist
Angel Jagger Makeup Artist
RedrumColl... Photographer
Chad Hess Photography Photographer
Marilyn San German Makeup Artist
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text:
|Publication number
|Publication type
|Publication date
|Oct 16, 1973
|Filing date
|Apr 21, 1972
|Priority date
|Apr 21, 1972
|Publication number
|Kielman F
|Original Assignee
|Sperry Sun Well Surveying Co
|Export Citation
|BiBTeX, EndNote, RefMan
|Patent Citations (5), Referenced by (34), Classifications (15), Legal Events (4)
|External Links: USPTO, USPTO Assignment, Espacenet
1451 Oct. 16, 1973 United States Patent 1191 Kielman, Jr.
l75/4.5l X Alberts 175/451 175/44 CIRCULATING SLEEVE 2,361,204 171323 Howard etal.
[75] Inventor: Fred 3,450,216 6/1969 l-lugeletal.
[73] Assignee: Sperry-Sun WellSurveying Company, Sllgarland,T X- Primary Examiner-David H. Brown [22] Filed, Apr 21 1972 Attorney-George L. Church et a1.
Applv No.: 246,293
[57] ABSTRACT In a drilling system, a subassembly provides a means for seating an instrument housing in the drill strin during a drilling operation and for permittin flowof drilling fluids around the instrume [52] US Cl 175/320, 175/45 [51] Int. E2lb l7/00 g a normal nt housing.
[58] Field oi Search...................... l75/4.5l, 44, 45, 175/73, 75, 61, 320, 324
The seat also provides means for orienting the instr ment housing in a predetermined position relative to a [56] References cued portion of the drill system.
UNITED STATES PATENTS 5 Claims, 3 Drawing Figures PATENIEnum 16 I975 3 765 494 sum 1 OF 2 y In l \3 PATENTED OCT 16 I373 SHEET 2 OF 2 FIG. 2
BACKGROUND OF THE INVENTION The present invention pertains to a circulating sleeve and more particularly to an apparatus for orienting an instrument within a drill string and providing for the circulation of fluid about the instrument tov permit drilling when an instrument is seated therein.
When drilling boreholes in the earths surface it is often desirable for various reasons to deviate the holes from a vertical course through the earth formations. The term directional drilling is applied to such operations. One example of the use of such directional drilling operations is found in the drilling of oil wells from offshore platforms. It is a common practice to build large drilling platforms which are permanently secured to the ocean floor and from which a multiplicity of wells are drilled. Because of the number of wells which are drilled from a single platform, it is necessary to drill the holes laterally away from the platform so that earth formations containing petroleum reservoirs may be penetrated at distances laterally spaced from the platform. This procedure permits production from as great an area as possible from a single platform. It is easily understood how important the maintenance of direction and dip of such boreholes is in order to penetrate particular formations at predetermined depths and thereby intersect the desired petroleum reservoirs.
One technique for obtaining such information as to the direction of a borehole is to cease drilling and run a surveying instrument into the drill pipe on a wire line. Alternatively, the instrument may be go-deviled to the bottom of the drill pipe. The instrument is oriented with respect to the drill stem by means of a muleshoe sub located at the lower end of the drill stem. The muleshoe" is simply a device for capturing the tool and orienting the tool in a predetermined radial direction with respect to a position on the drill ste'm. For example, the drill stem normally used in such'a directional drilling operation includes what is normally termed a bent sub," at its lower end which places a bend in the lower end of the stem and thereby permits angular deviation of the drill bit. The muleshoe" is normally oriented with respect to the bent sub. This in turn orients the instrument which is being positioned in the lower end of the drill stem. This series of orientation procedures provides a correlation between the direction in which the drill bit is angled and the directional alignment of the survey instrument. After the survey instrument is operated to generate a record, generally be means of a timing mechanism, the tool is retrieved to the surface by means of a wire line. The record is then processed and examined to determine the direction in which the hole is being drilled. a
More recently, techniques have been developed f passing an instrument into the drill stern by means of a conductor cable to provide continuous directional information to the surface during the drilling operation. This technique also utilizes the method described above of locating the instrument within a muleshoe which in turn is oriented in a particular radial direction with respect to a bent sub" in the drill string. Thus, the information which is received at the surface indicative of the direction in which the survey instrument is positioned is correlated with the direction that the bit is angled.
It is common in such directional drilling operations to use a mud motor or turbine for rotating the drill within the borehole. Such a motor or turbine requires the continuous circulation of drilling fluidsthrough the motor to provide the power for turning the drill. Therefore, it is essential that a constant flow of drilling fluids be maintained through the system even while an instrument is positioned within the drill stem.
It is therefore an object of the present invention to provide a new and improved circulating sub for receiving an orientation instrument within a drill string.
SUMMARY OF THE INVENTION With this and other objects in view, the present invention contemplates a drill pipe having a recessed portion therein forming a seat within the bore of the pipe. A sleeve which is sized for reception within the recessed portion has a shoulder which when received within the recess mates with the seat. A plurality of Iongitudinal ribs are formed on the sleeve and extend upwardly from the shoulder. Means extend inwardly from the sleeve for engaging a mating surface on an instrument housing to thereby locate the instrument in a predetermined manner within the sleeve.
BRIEF DESCRIPTION OF THE DRAWINGS DESCRIPTION OF THE PREFERRED EMBODIMENTS Referring first to FIG. 1 of the drawings, the lower end of the'drill stem is shown including a drill collar 27 and a muleshoe orienting sub 28. A bent sub 29, mud motor'or turbine 31, and a rotating bit sub 32 are positioned below the orienting sub. An instrument which for the purpose of this disclosure shall be referred to as an orienting tool 33, is shown positioned within the interior bore of the drill collar and is connected at its upper end with a conductor cable 26 extending to the surface. Typically, the conductor cable would extend through a line wiper at the surface and be taken up on a motor driven drum which permits its playout and take up during operations. The line wiper is constructed to seal off well pressure at the wellhead during line operations in the drill pipe.
Referring again to the lower end of the drill stern, a
muleshoe sleeve 34 is shown positioned within the muleshoe orienting sub 28 and is oriented therein in a predetermined fashion. The sleeve 34 is held in the predetermined position within the sub by means of a screw or the like 36 extending through the side wall of the sub 28. The muleshoe sleeve 34 is shown having a key 37 positionedin its sidewall and extending inwardly into the sleeve bore. The muleshoe and its key are normally oriented directionally with respect to the bent sub and thus with respect to the drill bit which derives its direction of inclination by means of the bent sub. This predetermined alignment of the muleshoe key with respect to the bent sub is convenient for purposes of determining the original orientation of the drill with respect to surface indications of tool direction and thereafter for making compensating changes in drilling direction. The muleshoe sleeve or circulating sleeve as it will be hereinafter referred to, may be utilized to positionally locate various types of instruments within a drill 'pipe; however, since one of its primary uses is for locating directional drilling orienting instruments this disclosure will continue to use such an instrument as an example.
The orienting tool 33 which is shown schematically in FIG. 1 includes a muleshoe 39 which is secured to the lower end of the tool string. The muleshoe includes a protruding shaft or stinger 41 having a tapered end 42 (see FIG. 2) for guiding the stinger into the muleshoe sleeve 34. A shoulder 43 extends spirally around'opposite sides of the stinger, meeting at a pointed terminus 44, to form a camming surface. On the opposite side of the tool from the terminus 44, the spiral shoulders 43 meet to form a short longitudinally extending slot 45. The slot 45 is sized to receive the inwardly extending key 37 on the muleshoe sleeve 34 when the tool 33 is positioned in the drill pipe. The tool string 33 further includes a muleshoe adjuster which permits rotation of the muleshoe 39 relative to the tool string 33. The adjuster includes includes mating portions (not shown) between the muleshoe and tool string to permit relative rotation therebetween, and a locking collar 35 for securing the tool string and muleshoe in a fixed rotational position. Greater detail of the muleshoe and its functional use with an orienting tool is shown in co-pending U.S. Pat. application Ser. No. 86,877, now U.S. Pat. No. 3,7l8,l94, which is assigned to same assignor as the present application.
Referring next to FIG. 2 of the drawings, the circulating sleeve 34 and its cooperative relationship with the sub 28 is shown in greater detail. Sub 28 has a recessed bore portion 46 with a tapered seat 47 at its lower end. The circulating sleeve 34 is shown positioned within the sub 28. The sleeve 34 has a body portion 49 with longitudinal ribs 48 extending outwardly therefrom into contact with the recessed inner bore portion 46 of the sub 28. The longitudinal ribs 48 extend downwardly below the lower end of the body portion 49. A cylindrical sleeve 51 is formed at the bottom end of the longitudinal ribs. Tapered top and bottom surfaces 52 and 53, respectively, are formed on the sleeve 51. The lower surface 53 is arranged to matingly seat on the tapered surface 47 of the recessed bore 46 in sub 28. Upper and lower peripheral grooves 54 and 56, respectively, are formed on the surface of the exterior wall of sleeve 51, and are sized to receive O-ring seals therein. Another peripheral groove 57 is formed between the grooves 54 and 56 in the outer wall of the sleeve 51 for receiving an inwardly extending end portion of a set screw 36. The body portion 49 of the circulating sleeve 34 and one of the longitudinal ribs 48 have an'aligned opening formed through the rib and body portion for receiving the key 37. The key 37 has an enlarged shoulder portion 61 formed thereon which abuts a complimentary shoulder formed in the rib opening.
In the operation of the system described herein the following method is employed: the circulating sleeve 34 is inserted into the upper end of sub 28 until the lower tapered surface 53 on sleeve 51 is matingly seated on tapered surface 47 within the bore of sub 28. The circulating sleeve is then rotated within the bore of sub 28 until the key 37 which extends inwardly from the sleeve is positioned in accordance with a predetermined arrangement to permit orientation with respect to a portion of the drill stem. Normally, the predetermined alignment of the key 37 is in some relationship with the bent sub 29 so that it is known that the muleshoe key 37 is pointing in a particular direction with respect to that in which the drill is pointing. This arrangement provides means for determining, by means of an orienting instrument to be located within the muleshoe sub in what direction the drilling operation is being conducted. In any event, after the key 37 and the circulating sub has been oriented within the sub 28, set screws 36 are rotated until they extend inwardly into the groove 57 on lower sleeve 51 and tightened therein to hole the sleeve in a fixed relationship with the sub 28 and thereby prevent its rotation within the sub. Next, the sub 28 is made up in the drill string and the drill string is lowered into the wellbore for purposes of performing the drilling operation.
Thereafter, when it is desired to lower an instrument into the drill stem by means of the cable 26 shown in FIG. 1, such instrument is lowered until the muleshoe stinger 41 extends into the circulating sub 34 whereupon the spiral camming surface 43 thereon engages the key 37 and produces a camming action to rotate the muleshoe until the longitudinal slot 45 of the muleshoe drips over the key 37. The procedure thus far described permits the instrument housing located above the muleshoe to be radially positioned within the drill stem in a predetermined manner. The arrangement of the circulating sub, seat and muleshoe permits the circulation of drilling fluids through the drill stem even when the instrument and muleshoe are seated within the drill string. This is provided by means of the annular spaces between the longitudinal ribs 48 and between the sleeve 51 and the lower end of the muleshoe. The path of such fluid circulation is shown by the arrows in FIG. 2.
With the circulating sub described herein the volume of fluid that may be circulated through the system is sufficient to drive the mud motor or turbine during drilling operations while permitting the continuous use of an orienting instrument.
Wile a particular embodiment of the present invention has been shown and described, it is apparent that changes and modifications may be made without departing from this invention in its broader aspects and therefore the aim in the appended claims is to cover all such changes and modifications as which fall within the true spirit and scope of this invention.
What is claimed is:
1. In a drilling system utilizing a hollow conduit for carrying a drilling fluid means for seating an instrument housing within the conduit while performing a drilling operation, which means includes: a seat formed within the bore of said conduit, a sleeve sized for reception within the bore of said conduit, shoulder means on one end of-said sleeve for providing a mating reception of said sleeve within said seat, said shoulder means being sized for close fitting reception within said conduit, a plurality of longitudinal ribs on said sleeve extending above said shoulder means and key means extending inwardly from said sleeve for orienting an instrument housing.
2. The apparatus of claim 1 and further including a transverse opening extending through one of said ribs and sleeve for receiving said key means.
3. The apparatus of claim 1 and further including means of locking said sleeve against relative movement with said conduit.
4. The apparatus of claim 3 wherein said locking means extends through the wall of said conduit and further including seal means on said shoulder arranged to be positioned above and below said locking means when said sleeve is seated in said conduit.
5. A circulating sleeve for seating and orienting an instrument housing within the bore of a drill pipe during a drilling operation and arranged to permit the flow of fluids through the drill pipe when an instrument housing is positioned therein, comprising: a sleeve for reception within the bore of the pipe, and having a cylindrical portion with longitudinal ribs formed thereon for engaging the bore of the pipe and maintain said cylindrical portion in a spaced position from said bore; a shoulder formed on said sleeve for engaging a seat in the pipe; key means extending from the wall of said cylindrical portion into its bore for engaging a portion of the instrument housing to orient the housing with respect to said sleeve; and means on said sleeve for receiving a member extending from the wall of the pipe for holding said sleeve against rotation relative to the pipe.
|Cited Patent
|Filing date
|Publication date
|Oct 17, 1938
|Apr 16, 1940
|Strength Thomas C
|Directional well drilling tool
|Oct 30, 1958
|Sep 4, 1962
|Eastman Oil Well Survey Co
|Apparatus for orienting well drilling equipment
|Jun 25, 1965
|Jan 2, 1968
|Pan American Petroleum Corp
|Method and apparatus for treating an underground formation
|Sep 26, 1966
|Dec 3, 1968
|Halliburton Co
|Oriented perforate test and cement squeeze apparatus
|May 21, 1965
|Jun 17, 1969
|Christensen Diamond Prod Co
|Core orienting apparatus and method
|Citing Patent
|Filing date
|Publication date
|Jul 1, 1977
|Dec 19, 1978
|Wilson Industries, Inc.
|Flow-through mule shoe sub
|May 4, 1978
|Jan 1, 1980
|Scientific Drilling Controls, Inc.
|Remote well signalling apparatus and methods
|Mar 8, 1982
|Oct 29, 1985
|Exploration Logging, Inc.
|Apparatus for well logging telemetry
|Sep 25, 1987
|Dec 6, 1988
|Rehm William A
|Orienting and circulating sub
|May 27, 1997
|Mar 16, 1999
|Pegasus Drilling Technologies L.L.C.
|Bottomhole assembly orienting sub
|Jul 24, 2006
|Nov 24, 2009
|Halliburton Energy Services, Inc.
|Steerable drilling system and method
|Apr 25, 2007
|Feb 9, 2010
|Ethicon Endo-Surgery, Inc.
|System and method for determining implanted device orientation
|Mar 7, 2006
|Aug 17, 2010
|Ethicon Endo-Surgery, Inc.
|System and method for determining implanted device positioning and obtaining pressure data
|Mar 7, 2006
|Aug 17, 2010
|Ethicon Endo-Surgery, Inc.
|Non-invasive pressure measurement in a fluid adjustable restrictive device
|Feb 7, 2008
|Nov 30, 2010
|Ethicon Endo-Surgery, Inc.
|Powering implantable restriction systems using light
|Jan 29, 2007
|Apr 19, 2011
|Ethicon Endo-Surgery, Inc.
|External mechanical pressure sensor for gastric band pressure measurements
|Mar 7, 2006
|Sep 13, 2011
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|Dec 14, 2007
|Jan 24, 2012
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|Adjustable height gastric restriction devices and methods
|Feb 8, 2008
|Feb 14, 2012
|Ethicon Endo-Surgery, Inc.
|System and method of sterilizing an implantable medical device
|Dec 27, 2007
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|Ethicon Endo-Surgery, Inc.
|Controlling pressure in adjustable restriction devices
|Feb 28, 2008
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|Physiological parameter analysis for an implantable restriction device and a data logger
|Mar 6, 2008
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|Dec 10, 2007
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|Methods for implanting a gastric restriction device
|Jan 28, 2008
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|Methods and devices for measuring impedance in a gastric restriction system
|Feb 7, 2008
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|Ethicon Endo-Surgery, Inc.
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|Methods and devices for diagnosing performance of a gastric restriction system
|Dec 27, 2007
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|Constant force mechanisms for regulating restriction devices
|Jan 28, 2008
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|Gastric restriction device data handling devices and methods
|Feb 12, 2008
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|Ethicon Endo-Sugery, Inc.
|Automatically adjusting band system
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|GUI for an implantable restriction device and a data logger
|Jul 24, 2006
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|Chen Chen-Kang D
|Steerable drilling system and method
|Mar 31, 2006
|Oct 4, 2007
|Macpherson Calum Robert
|Drill string system for performing measurement while drilling and logging while drilling operations
|Feb 20, 2015
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|Aps Technology, Inc.
|Pressure locking device for downhole tools
|May 22, 1998
|Dec 3, 1998
|Pegasus Drilling Technologies, L.L.C.
|Bottomhole assembly orienting sub
|U.S. Classification
|175/320, 175/45
|International Classification
|Cooperative Classification
|European Classification
|May 7, 1992
Owner name: BAROID CORPORATION, TEXAS
Free format text: RELEASED BY SECURED PARTY;ASSIGNOR:CHASE MANHATTAN BANK, THE;REEL/FRAME:006085/0590
Effective date: 19911021
|Jun 15, 1989
Owner name: BAROID TECHNOLOGY, INC., A CORP. OF DE., DELAWARE
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST.;ASSIGNOR:SPERRY-SUN DRILLING SERVICES, INC., A CORP. OF DE.;REEL/FRAME:005208/0161
Effective date: 19890613
Owner name: SPERRY-SUN DRILLING SERVICES, INC.
Free format text: CHANGE OF NAME;ASSIGNORS:NL ACQUISTION CORPORATION, (CHANGED TO);SPERRY-SUN, INC., (CHANGED TO );NLSPERRY-SUN, INC., (CHANGED TO);REEL/FRAME:005208/0157
Effective date: 19810421
Owner name: SPERRY-SUN, INC., A CORP. OF DE., DELAWARE
Free format text: CHANGE OF NAME;ASSIGNOR:SPERRY-SUN WELL SURVEYING COMPANY;REEL/FRAME:005208/0153
Effective date: 19760617
|Mar 8, 1989
Owner name: CHASE MANHATTAN BANK (NATIONAL ASSOCIATION), THE
Free format text: SECURITY INTEREST;ASSIGNOR:BAROID CORPORATION, A CORP. OF DE.;REEL/FRAME:005196/0501
Effective date: 19881222
|Feb 14, 1988
Owner name: BAROID TECHNOLOGY, INC., 3000 NORTH SAM HOUSTON PA
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST.;ASSIGNOR:SPERRY-SUN DRILLING SERVICES, INC.;REEL/FRAME:005024/0898
Effective date: 19890210
Owner name: SPERRY-SUN DRILLING SERVICES, INC.
Free format text: CHANGE OF NAME;ASSIGNOR:NL SPERRY - SUN, INC.;REEL/FRAME:005024/0939
Effective date: 19880214
Owner name: SPERRY-SUN, INC.
Free format text: CERTIFICATE OF INCORPORATION TO RESTATE INCORPORATION, EFFECTIVE JULY 21, 1976;ASSIGNOR:SPERRY-SUN WELL SURVEYING COMPANY;REEL/FRAME:005024/0918
Effective date: 19760617


text:
Thursday, January 24, 2013
the buzz: Jan 24
This week's news has got food bloggin written all over it. That's right people. The details for the Food Bloggers of Canada 2013 Conference is out and tickets go on sale tomorrow!!! Other than it being the first event of its kind in Canada (incredible right?), there are loads of incredible speakers and workshops and even goodies to take home. Plus, I'll be there so what are you waiting for? For you local bloggers, I'm having a lil' food blogger meet up/potluck in February to chat about it so get at me if you want in.
What's bubbly and hoppy and refreshing? You got it. Beer. And lucky for us in Hali a new microbrewery has joined in the beer game. Bridge Brewing opened yesterday at 2576 Agricola after months of hard work by owner Peter Burbridge. Folks were so eager to get their hands on the brew that they sold out in three hours. Yup, three. Lucky for us, they were fully stocked when we dropped by to check it out today. The space is rad, the beer is delish (I may have tried it before! Shh!), even the bottles are cool. So drop by, get yourself a growler and be prepared to go back the next day for a refill.
Well, out here on the East is pretty damn cold. The only thing getting me through it, other than hot baths and my couch blankie (yeah, I have a blankie, so what!?), is a February full of food and drink. Like I mentioned last week, next month will bring us both the Winter Wine Festival and the Savour Food and Wine Festival. New deets are out for both so click your way over to their sites. In particular, check out the Dine Around menus. Last year we had an incredible post Valentine's day dinner at Chives, taking advantage of the sweet three course Dine Around deal. Their menu this year is looking equally as delish but I have a feeling we'll be checking out somewhere new. Although there are 28 days in February...
Lastly, today I finally made my way through the entire Ace Burger Co. menu after demolishing a Jerk Chicken. Another party in my mouth. If you haven't been, you really gotta check it out. Truly, the best damn burgers in the city. GET SOME.
Until next time.
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Kenny ChesneySat, May 18, 2013 - 5:00 PMMiller Park in Milwaukee, WI
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The Center for Women, Politics & Policy (CWPP)
Human Trafficking in the Portland Metro Area
A Panel Discussion
Wednesday, November 17, 2010
Urban Center, Room 212
Portland State University
506 SW Mill St.
Portland, OR 97201
Confirmed panelists include:
Concetta Schwesinger
Oregon Women Lawyers, Oregon Department of Jusice
Yonsoo Lee
Trafficking Investigator with the FBI
Esther Harris
Youth Development Coordinator, Salvation Army White Shield Center
Light complimentary refreshments will be included. No cost for participation.
Please RSVP by emailing email@example.com with any further questions.
The evening will begin with a 40-minute panel of professionals who have worked closely on this issue. Then we will identify the legislative, enforcement and preventive components in an attempt to better understand this significant human rights issue. The evening will conclude with a discussion of our findings.


text:
TONIGHT: Through the Ropes!
Boxing fans, get ready to climb "Through the Ropes" with the #1 boxing news radio show hosted by award winning boxing writer Marc Lichtenfeld! This week, the most feared fighter in boxing, Paul "The Punisher" Williams climbs Through the Ropes. Fightnews.com's Flattop will also be on the show. Call or email your questions to email@example.com.
Through The Ropes airs live coast to coast on Thursday nights from 9:00 – 10:00 ET on 75 affiliates of the Sports Byline USA Broadcast Network, including great radio stations like KSHP 1400 AM in Las Vegas and WNSR 560 AM in Nashville. The one-hour show will also be heard on American Forces Network, Sportsbyline.com and Fightnews.com. Fightnews readers can listen live by clicking tonight at 9:00 p.m. ET.


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ED Magedson – Founder
Ripoff Report
Government Grants Association -- Brian Johnson -- Melvin Smith (both Sound Middle Eastern Accent)4100 Eastbay Dr. Suite B-40 Clearwater, Florida U.S.A.
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This report was posted on Ripoff Report on 10/29/2004 05:13 PM and is a permanent record located here: http://www.ripoffreport.com/reports/government-grants-association-brian-johnson-melvin-smith-both-sound-middle-eastern-accent/clearwater-florida-33764/government-grants-association-brian-johnson-melvin-smith-both-sound-middle-eastern-acc-115280. The posting time indicated is Arizona local time. Arizona does not observe daylight savings so the post time may be Mountain or Pacific depending on the time of year.
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Here you go...
Now let the posts with silly fight results begin...
Here you go...
Now let the posts with silly fight results begin...
SURGEON GENERAL'S WARNING : Quitting 2-Stroking Now Greatly Reduces Serious Risks to Other People's Health.
up all night writing papers....., couldnt resist a little procastination
suzuki beat honda in a split decision: 15 300 000 to 13 700 000
ron jeremy 2,600,000 vs your mom 23,800,000
looks like your mom can take ron jeremy
Ah just what i thought....
Masterbation = 846,000
Anal Sex = 9,580,000
soft: 75,300,000 hard: 221,000,000
Lets Jump
Erm, I did the same fight on my own. Honda beat Suzuki...?Originally Posted by Fusilli Jerry
dont forget my gas prices vs consumers fight
hmmm I dont think so...
Last edited by Good old Pete; 04-04-2009 at 09:05 PM.
When speeding run from the cops, it's a lesser infraction.
But CSB has porn!!


text:
Earnings flat, Kohl's sees tougher 2008
February 29, 2008,
Menomonee Falls, Wis. – While core basics left much to be desired in sales, it was the name branded goods, like Food Network, Simply Vera Vera Wang and Chaps, that proved "exceptionally strong" in the home department in 2007 at Kohl's stores, the company said in its fourth quarter and yearend earnings call late yesterday.
For the fiscal year ended February 2, 2008, net income was $1.1 billion, or $3.39 per diluted share, compared with $1.1 billion or $3.31 per diluted share, a year ago. Margins were cramped, as net sales of $16.5 billion were up 5.6% over $15.6 billion a year ago. On a comparable 52-week basis, comp sales fell 0.8%.
Home, along with women's apparel, "trailed the company in both periods," said Kohl's president Kevin Mansell of the quarterly and annual results.
Looking closer at home furnishings, he said, "There was no particular category that did much worse than any other, and from a brand perspective, almost the exact same thing is true."
Mansell said that in home, much of the "core basics had a tough falling out," while recently introduced Food Network-branded products were "exceptionally strong" and the Simply Vera Vera Wang and Chaps lines showed "positive" results.
"The customer continues to respond well to all our new brand launches and their penetration continues to increase with substantial room to grow," he continued.
In Kohl's initial guidance for fiscal 2008, the company assumes a total sales increase of 5% to 8% and a comp sales change of 0% to negative 3%, and projects earnings per diluted share of $3.15 to $3.50 for the year. For the first fiscal quarter, Kohl's expects earnings per diluted share of $0.50 to $0.54.
The home-related brand launches for 2008 include Jumping Beans, an opening price point assortment in children's goods, which debuted in February; and a mostly hardlines program of Bobby Flay-branded kitchen products, crafted with the celebrity chef as part of Kohl's Food Network platform, to roll out in May.
On the marketing and advertising front, Kohl's this year intends to "dramatically expand" its "versioning" of materials to what Mansell said would "reflect the differences by individual market across the country, be it lifestyle or climate."
The new store count for 2008 calls for 70 to 75 units. Of these, 28 will open this spring and the balance – including Kohl's 1,000th store – will open in the fall, when Kohl's plans to enter the Miami/Ft. Lauderdale metro markets for the first time, with eight units.
In 2007, by comparison, the retailer opened 112 stores.
During the question-and-answer session of the earnings call, an analyst asked Kohl's top executives to comment on competition between its Chaps program and JCPenney's new American Living line, created by Polo Ralph Lauren.
"Chaps had a very good year at Kohl's, a very solid fourth quarter, particularly in men's, and we expect to get off to good start this year," said Larry Montgomery, ceo. Mansell added that Chaps "is a brand that has been around for a lot of years; the customer knows it well…and we are giving spectacular value for high-quality from a great sourcing customer."
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Chapter Fifteen, Part Two: The Temptation of Saint Anthony, 1945Mature
Her Koschei drags his foot back, the toe of his converse bumping like a skipped stone on the grates as he moves to kick her again.
The Doctor is the only thing holding him back, quite literally, because without his steadying hand, the other Time Lord would slip to the grates again. He's weak for some reason. The two of them, together... at the party, too. It does make sense. But the Doctor is affected, so it must have been the Master who parallel parked the TARDIS near the exit.
"…you disguised a TT capsule as a storage cupboard?" she asks, injecting incredulity while she looks around the console room, committing everything to memory.
"She won't let you fly her, sweet child," the Doctor says, pushing himself up. His tweed jacket is rumpled; his bowtie undone. He staggers to the console and just leans. "… she thinks you need a spanking. But that's all right; she thinks that about me most days."
He's looking at her as though he expects her to believe him, Flamina realizes as she shifts one knee out from under her rump and sits up a bit. She glares up at the Master, who is, strikingly, standing between the Doctor and herself. She wipes her chin with her white silk sleeve. "Why do you care about this animal, Koschei? He tried to burn us! He killed his own children. Why did you save him?"


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Recycled barrel pub table with tequila bottle holder, horse shoe and bowl stand. Exclusive, unique rustic wine furniture.Check out our stool category where you can purchase matching stools to this barrel table.
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Spectrum by Uzma Sajid is one of the new emerging fashion brand in our country. Spectrum by Uzma Sajid has introduced its Spectrum by Uzma Sajid Summer Dresses Collection 2013 for women and girls.Spectrum by Uzma Sajid Summer Collection 2013 consists of long shirts and frocks with trousers and churidar pajamas.Spectrum by Uzma Sajid has overstated this collection with embroidery and laces work.The colors used for this collection are bright such as red, green, maroon, yellow and etc.These dresses are perfect for every party and functions.Women and girls will find total stylish and stunning dresses which are perfect for any kind of party and functions.Let's see picture here below,
Tag Archives: women party wear collection
Silkasia Summer Party Wear Dresses Collection 2013
Silkasia Bridal and Formal Dress Collection 2013 For Women is a Pakistani fashion designers brand located & launched in Karachi in 2001. Reently, Silkasia launched its latest collection titled as Silkasia Bridal and Formal outfits 2013 For Women. Banarsi dress is a well known dress not in Pakistan but also all over the world.
Many dress designers have produce a lot of fashion in banarsi trend. Silkasia Banarsi trendy, chic chiffon throng 2013, formal, semi formal and promo ostensible attires. Buy Latest Pakistani Ladies Fashion Clothes, Shalwar Kameez, Anarkali, Pishwas, Casual, Formal, Party, Bridal & Wedding Wear Dresses with Delivery Round the Globe. Chiffon Brush Print Semi formal Collection In store now. Make your summer cool with Fine cotton & pure silk suits at SILKASIA.


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text:
The author of the Portus debacle was born in Israel, where his father, Daniel, developed defence systems for the Israeli military before moving his family to Canada in 1988. In Toronto, Daniel founded EIS Electronic Integrated Systems Inc., which manufactures traffic-monitoring equipment. The younger Manor had a classic Toronto Jewish upbringing, growing up in Forest Hill and attending Forest Hill Collegiate Institute. His yearbook photo for 1992, his graduating year, sported this oddly prescient and creatively spelled caption: "Into the future I see an array of precious jems, dimonds, and rubies glowing with effervesent light…and a guy named Louie yelling 'Put the jems in the yellow bag, the diamonds in the green bag, and get a move on it before we get busted.'"
Manor went on to attend the University of Toronto and completed a degree in applied science, graduating in 1996. But business was his calling. After working briefly for his father's firm, Manor met entrepreneur Michael Mendelson, who was running a small merchant bank, KBL Capital. Mendelson gave Manor some work, and soon they were fast friends, united by a desire to become spectacularly rich.
After KBL Capital took a hit when the dot-com bubble burst in 2001-'02, Manor and Mendelson looked around for a new line of business and settled on hedge funds. They devised an investment plan that was brilliantly simple: They told investors that all of their money was going to be plowed into principal-protected notes issued by a large bank–in this case, the French giant Société Générale. The bank then promised the notes would be linked to returns of a basket of hedge funds.
Normally, retail investors are not allowed to invest in hedge funds; they're too risky. But this structure provided a workaround. The upshot was that, after five years, investors would be guaranteed to get at least their principal back, plus whatever was made by being linked to the hedge funds. "You're selling zero downside," explains Mendelson. "And your upside is, you're tied to a sexy asset class." The duo's dream was to build an investment management firm that would grow to $5-billion in assets, sell it off and pocket $250-million apiece.
They opened Portus in January, 2003, with Mendelson acting as the operations guy and Manor the investment guru. Things took off when they managed to land a referral arrangement with a group of securities dealers, including a branch of Manulife Financial Corp., one of the most solid names in the financial industry. This meant hundreds of brokers across the country were steering their retail clients to Portus. It was like striking oil, with hundreds of millions coming through the door every quarter at one point. Eventually employing 120 people, Portus leased offices on the 24th floor of BCE Place at the foot of Bay Street, with a stunning view of Lake Ontario. It was a cocky, young workplace that "reeked of arrogance," recalls one former Portus salesman. "People wanted to work for us because we were the shit." Hours were long, expectations high. Vacations were frowned on.
In the office, Manor was introverted and private. "Boaz was often in his office with the door closed," says the former employee. "He was the strong, silent type. He was paranoid." Nevertheless, Manor was the face of Portus in marketing material and on a ghostwritten book, A Guide to Alternative Investing.
Despite appearances of success, Portus had a design flaw. Its managed account agreement said Portus would be entitled to an annual management fee ranging from 1.9 per cent to 2.25 per cent of the market value of assets in an investor's account. But Mendelson and Manor felt this was not enough money to grow the business as fast as they wanted. In particular, they needed a lot of cash to persuade financial planners to steer clients to Portus.
Their solution, as it turned out, was illegal. Although the firm's managed account agreement explicitly said Portus was required "to invest all assets which the investor contributes," in reality only 86 cents of every dollar was invested in the principal-protected notes. The remaining 14 cents went to meeting Portus's payroll and expanding the business by paying fees to brokers. "That was the fraud," says Joel Vale, a Toronto lawyer involved in a lawsuit against Portus's auditors, PricewaterhouseCoopers LLP, after the hedge fund crashed.
Mendelson claims it was not this straightforward: He says Portus received a huge discount from Société Générale, which Portus then used, through a complex manoeuvre, to pay for operating expenses. And Portus received a thumbs-up from its lawyers that it was perfectly legal. What no one disputes is that none of this was disclosed to investors, as it should have been.Report Typo/Error
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