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A critical directive of the BioMicro Center is to provide a cutting-edge research core for members of the MIT community. Creating and maintaining the BioMicro Center at the forefront of technology improves our ability to support MIT faculty in grant applications, manuscript publishing and in the recruitment of new faculty members. In part, this goal is achieved through ongoing collaborations with many labs at MIT. A selection of these collaborations taken from the annual reports of the BioMicro Center is presented below.
- 1 2013
- 2 2012
- 3 2011
- 4 2010
Copy number analysis in single cells - AMON LAB- Biology and KI
Accurate segregation of chromosomes is critical for maintenance of genomic stability. Failure to properly segregate chromosomes results in an unbalanced chromosome number, a condition known as aneuploidy. Aneuploidy for chromosome 21 is the cause of Down Syndrome, and the majority of solid tumors harbor aneuploidies for one or more chromosomes. Apart from these diseases, it has also been reported that normal, untransformed cells missegregate chromosomes as often as 20% of divisions (Carere et al., 1999; Shi & King, 2005). Furthermore, multiple groups have reported even higher levels of aneuploidy in the normal mammalian brain, with over 20% of neurons and glia being aneuploidy (Faggioli, Wang, Vijg, & Montagna, 2012; Pack et al., 2005; Rehen et al., 2001; 2005). Traditionally, these phenomena have been studied using spectral karyotyping (SKY) and fluorescence in situ hybridization (FISH). Unfortunately, these methods are susceptible to spreading and hybridization artifacts, respectively, and FISH is limited in the number of chromosomes that can be surveyed in a single experiment. The Amon laboratory has thus turned to single cell sequencing to provide a more robust and thorough assessment of aneuploidy in the brain and other tissues.
Using tissue samples obtained from mice and humans, we isolated single cells by microaspiration and amplified genomic DNA using a commercially available kit. We then relied on the BioMicro Center to prepare barcoded libraries and sequence these libraries using the Illumina HiSeq2000. From sequencing reads, we were able to infer the copy number of each chromosome in a given cell.
Cell cycle control in S. meliloti - WALKER LAB - Biology, CEHS, and KI
Proper regulation of the bacterial cell cycle is crucial for faithful replication and segregation of the genome. In some groups of bacteria, i.e. α-proteobacteria, tight regulation of cell cycle progression is also necessary for specific cellular differentiation required for adaptation to their diverse environmental niches. The symbiotic lifestyle of the soil bacterium, Sinorhizobium meliloti requires a drastic cellular differentiation that includes changes to the cell envelope, endoreduplication of the genome and loss of reproductive capacity. This differentiation requires a specific re-wiring of the S. meliloti cell cycle network to allow for the de-coupling of DNA replication and cell division. Although many of the well-characterized genes encoding the cell cycle regulatory network of the model α-proteobacterium, Caulobacter crescentus, are well conserved in S. meliloti, the function of these genes S. meliloti is poorly understood. It has been postulated that transcriptional control of cell cycle progression is a common and critical level of regulation in α-proteobacteria, but has only been demonstrated in C. crescentus. The BioMicro Center worked closely with Nicole De Nisco of the Walker lab work to perform global analysis of cell cycle regulated gene expression in S. meliloti, the first analysis of this kind conducted in an α-proteobacterium other than C. crescentus.
Taking advantage of the S. meliloti cell synchronization method developed by Nicole De Nisco, the BioMicro Center and Nicole De Nisco utilized custom Agilent gene expression microarrays to measure gene expression in S. meliloti cells isolated during discrete cell cycle stages. Through statistical analysis and data clustering executed by the BioMicro Center informatics staff, the Walker lab was able to define a set of 462 genes with cell cycle transcripts in S. meliloti, confirming the conservation of cell cycle regulated gene expression between S. meliloti and C. crescentus and thereby expanding the paradigm of transcriptional control of α-proteobacterial cell cycle progression. Using BLAST and COG analysis performed by the BioMicro Center to compare the S. meliloti list to the list of 553 previously identified genes with cell cycle regulated transcripts in C. crescentus, the Walker Lab discovered a core set of 128 genes with conserved cell cycle dependent transcription between the two species completely independent of cell synchronization method.
To determine which S. meliloti cell cycle regulated transcripts could be controlled by the transcriptional regulators CtrA and DnaA, the Walker Lab and BioMicro Center utilized previously described PWMs to search for CtrA and DnaA binding motifs within the list of 462 cell cycle regulated genes. This analysis revealed 64 genes with putative CtrA binding sites and 96 with putative DnaA binding sites. To provide support for the biological relevance of these sites, homologs of the S. meliloti genes identified in this analysis were found in 11 representative α-proteobacteria and then analyzed for conserved CtrA and DnaA binding motifs. The results of this analysis revealed a role for CtrA in regulation of cell division, flagellar biogenesis and coordination of DNA replication in S. meliloti, but by different strategies than previously defined in C. crescentus reflecting unique lifestyles and environmental niches of these two α-proteobacterial species. This lack of conservation of DnaA binding motifs in the homologs of S. meliloti genes demonstrating cell cycle regulated transcription revealed that the activity of DnaA as a transcriptional regulator in α-proteobacteria might be much less evolutionarily constrained than that of CtrA.
Genomics of Toxoplasma gondii - SAEIJ LAB - Biology
The overall goal of the Saeij lab is to understand the molecular basis for individual differences in outcome of infection. We believe that these differences are either due to host differences in resistance/susceptibility to infection or to genetic differences in the pathogen that causes the infection. As a model we use infection with the obligate intracellular pathogen Toxoplasma gondii. Toxoplasma can infect all warm-blooded animals and a third of the world's population is estimated to be infected. There are many different Toxoplasma strains that differ hugely in virulence in both mice and humans. Mouse and rat strain differences in resistance/susceptibility to Toxoplasma have also been described.
To characterize Toxoplasma genetic diversity we sequenced (Illumina HiSeq), with the help of the BioMicro Center, the whole genome of 10 Toxoplasma strains and the transcriptome of 32 different strains, representing global diversity. This data allowed us to construct the first Toxoplasma haplotype map and to propose a new model explaining current global diversity. The results were published in PNAS (Minot et al, 2012). We also determined the transcriptome of murine macrophages infected with these 32 different strains. This approach allowed us to correlate genotype with phenotype and have led to the identification of Toxoplasma loci and genes that affect fitness, clonality, virulence and modulation of host signaling pathways. Some of these results were published in PLoS Pathogens (Niedelman et al, 2012).
To determine mouse genetic differences in resistance/susceptibility to Toxoplasma we determined the transcriptome and toxoplasmacidal activity of naïve or IFNγ+TNF-stimulated macrophages isolated from 29 recombinant inbred mice, derived from A/J (Toxoplasma resistant) and C57BL/6 (susceptible) mice. We then identified mouse genomic loci affecting mouse gene expression and toxoplasmacidal activity using quantitative trait locus (QTL) analysis. We are now using genetic approaches to confirm mouse candidate genes involved in macrophage differences in Toxoplasma killing ability. The results of this study have helped us secure four years of grant support from the New England Regional Center of Excellence Biodefense and Emerging Infectious Diseases.<BE><BE> We have received excellent help from the BioMicro Center at every step of these analyses, their many suggestions on how to analyze the data have been instrumental in getting high quality data.
Sequencing of tRNAs - DEDON/NILES LABS - BE and CEHS
Emerging evidence points to selective translation of critical proteins as a major facet of the cellular stress response. The Dedon lab has shown that while one contribution to this translational control of cell response involves changes in the relative quantities of dozens of modified ribonucleosides in tRNA, there is also evidence for control of the number of copies of individual tRNA molecules. This dual control problem makes it difficult to distinguish changes observed in the level of tRNA modifications caused by altered activity of ribonucleoside-modifying enzymes from changes in the number of tRNA copies. To solve this problem, the Dedon lab has undertaken the development of a high-throughput deep-sequencing method to identify and quantify individual species of tRNA in a bulk population of tRNAs. This approach exploits the paucity of ribonucleoside modifications at the 3'-end of tRNA molecules, modifications that can interfere with RNA sequencing reactions. Following ligation of a custom primer to the 3'-ends of tRNA molecules, reverse transcription is performed to create cDNA that is subsequently subjected to linear amplification off of the custom primer. The amplified products are then identified and quantified by Illumina sequencing. This method allows for the simultaneous quantification of individual tRNA species in total tRNA isolated from cells subjected to different stresses, with application to study controlled degradation of tRNA during cell responses and to differentiate between changes in tRNA copy numbers from changes in RNA modifications due to enzymatic activity.
The BioMicro Center has been very instrumental in providing technical expertise towards optimization of experimental design for the conversion of tRNA to cDNA for sequencing, as well as assisting with bioinformatics analysis of the output data from the Illumina HiSeq 2000. The Dedon lab was able to barcode six different experimental samples so that multiple sequencing libraries could be run in a single lane of an Illumina flow cell. Using the Illumina HiSeq 2000, we were able to identify every tRNA species found in S.cerevisiae and are in the process of optimizing the sequencing method in order to reliably quantify relative changes in the levels of specific tRNAs isoacceptors from different populations of tRNAs.
The Niles lab has partnered with the Dedon Lab and the BioMicro Center in their efforts to develop a protocol for sequencing bulk tRNA. While the Dedon lab is investigating the effect of different stresses on tRNA expression, the Niles lab is interested in the basic endogenous biology of the malaria parasite. Currently, very little is known in the field about tRNA dynamics across the complicated life cycle of Plasmodium falciparum. While many published studies have tracked RNA expression using microarrays and deep sequencing, these protocols enrich for protein coding RNAs, excluding an important component of host biology. Additionally, while the mitochondria does not have any genes to produce tRNA, the apicoplast organelle has a full complement of tRNA genes. As the apicoplast organelle is required for successful parasite infection, having a better understanding of the expression of tRNA from the reduced but necessary apicoplast plastid will assist in the downstream search for both vaccines and pharmacological treatments.
Using the method described above, all tRNAs found in both the genome and apicoplast were detected from extracts of Plasmodium falciparum. However, there were several species of tRNA that jackpotted, appearing at concentrations several orders of magnitude above the remaining species. We are continuing to work with the BioMicro Center and the Dedon Lab to troubleshoot the protocol to determine a more quantitative picture of tRNA dynamics in the parasite.
Elucidating Brn targeting of Sox2 in embryonic stem cells - JAENISCH LAB - Biology and KI
In mammals, a few thousand transcription factors regulate the differential expression of greater than 20,000 genes to specify ~200 cell types during development. How this is accomplished has been a major focus of biology for many years. Transcription factors bind sequence-specific regulatory elements, including proximal promoters and distal enhancers, to control gene expression. Emerging evidence indicates that transcription factor binding at distal enhancers plays important roles in the establishment of tissue-specific gene expression programs during development. Combinatorial binding among groups of transcription factors can further increase the diversity and specificity of regulatory modules governed by a particular factor. The identification of regulatory modules comprised of groups of transcription factors which occupy important regulatory regions of genes which govern cell fate determination would shed light on how developmental decisions are made.
Work from the Jaenisch and Young labs elucidated the role of a group of three transcription factors, Oct4, Sox2, and Nanog, in regulating cell identity in embryonic stem cells (ESCs) using ChIP-on-Chip technology in 2005. Recently, Michael Lodato from the Jaenisch lab, in collaboration with the Boyer lab, has studied the genome-wide role of Sox2 in neural progenitor cells (NPCs). Their work showed that Sox2 occupied a distinct set of targets in NPCs relative to it targets in ESCs, and further that Sox2 switched partner factors during this transition: form Oct4 and Nanog in ESCs to Brn2, and Oct4 family member, in NPCs.
Taking advantage of the expertise in the BioMicro Center, they were able to examine the effect of the NPC-specific Sox2 partner factor, Brn2, on Sox2 binding in ESCs, where Sox2 normally partners with Oct4. The BioMicro Center Staff utilized the IP-Star Automated ChIP System to facilitate the automation of a large number of ChIP-Seq experiments, successfully querying the genome-wide occupancy of Sox2, Brn2, Histone H3 Lysine 4 monomethylation, Histone H3 Lysine 27 acetylation, total Histone H3, and p300 in both control and Brn2-induced ESCs in a rapid and controlled manner. Thus, not only could the binding of Sox2 and Brn2 be investigated, but due to the availability of the IP-STAR automated ChIP system the Jaenisch Lab could investigate changes to the epigenome as well. The BioMicro Center then prepared Illumina libraries from these samples ran them in a single lane of an Illumina flow cell, saving time and immensely reducing the cost to the Jaenisch Lab. Finally, the BioMicro Center implanted quality control metrics on both the samples before loading on the sequencer and on the quality of the data generated during the run.
Using this data, the Jaenisch Lab was able to define a set of regions where ectopic Brn2 could recruit endogenous Sox2 (Figure x), and they are currently investigating the effects of this binding on the chromatin status of these regions. The technical expertise of the staff of the BioMicro Center contributed significantly to the successful completion of this project during both the planning and execution of these experiments.
Large-scale discovery and functional analysis of distal enhancer elements - BOYER LAB - Biology and KI
The overall goal of the Boyer lab is to understand how a single cell can ultimately specify the diversity of cell types during mammalian development. An exciting and emerging area of biology in the post-genomics era has been the genome-wide identification of non-coding regulatory elements in what was once known as "junk DNA". Enhancers are key cis-regulatory elements that can affect gene transcription independent of their orientation or distance that are required for tissue specific patterning of gene expression during development, though only few examples had been known. Global identification of these regions as well as their contribution to target gene expression has been challenging because enhancers can often reside thousands of base pairs away from their target of regulation.
The Boyer lab has recently discovered that specific histone modification patterns could identify enhancers by genome-wide ChIP-Seq in embryonic stem cells (ESCs) as well as in a range of differentiated cell types and moreover, that these patterns distinguish enhancers as either active or poised (or inactive). Remarkably, genes connected to active enhancers code for genes with cell type specific functions and more importantly, poised enhancers could predict future developmental potential of that cell by marking genes that have the potential to become activated. However, it had been unclear how enhancer states were correlated during lineage commitment. Using cutting edge high-throughput sequencing methods, the Boyer lab has now defined a large set (~80,000) of both poised and active enhancers throughout the genome based on chromatin modification patterns derived from four key time points during cardiomyocyte differentiation. The differentiation system provides a unique opportunity to study enhancer state transitions during embryonic patterning of cardiomyocytes, which ultimately comprise the majority of the cell types in the developing heart.
The BioMicro Center was instrumental in providing the technical expertise necessary for the generation of the large number of high quality sequencing libraries from chromatin immunoprecipitated material. The BioMicro Center adapted the use of the IP-Star automated ChIP system (currently under evaluation) to facilitate automation of ChIP followed by library generation on the SPRI-TE. Additionally, the Boyer lab was able to barcode each experimental sample so that multiple sequencing libraries could be run in a single lane of an Illumina flow cell. Barcoded libraries were then analyzed by a number of quality control measures developed by the BioMicro Center to ensure the highest quality of sequence. These steps represented substantial improvements over previous protocols and allowed us to perform many experiments in a cost and time-efficient manner.
Together with the BioMicro Center, the Boyer lab analyzed the substantial amount of sequencing data and developed new algorithms to identify and to functionally dissect the role of distal enhancer elements in regulating gene expression patterns during lineage commitment. As a result of this study, they found that enhancer utilization is highly cell type specific and that enhancer state transitions are dynamic and non-random and likely occur during short windows of developmental time. These exciting findings have provided new details about how tissue specific expression patterns are established early in development and how mutations in these elements may contribute to cardiac diseases.
Identification of mutations in Caulobacter crescentus - LAUB LAB - Biology
A major challenge in bacterial genetics is the identification of the molecular targets and pathways affected by newly discovered genes. Toward this end, one powerful technique involves the unbiased selection for mutations that are able to suppress the deleterious effects of gain- or loss-of-function mutations in the gene of interest. However, finding the genomic locations of these suppressor mutations by traditional mapping methods can be time and labor intensive. The BioMicro Center has worked with the Laub lab to bypass the need for genetic mapping by sequencing the entire genomes of mutant bacterial strains.
The Laub lab, working in the bacterium Caulobacter crescentus, has recently characterized a novel gene, sidA, which inhibits cell division in response to DNA damage. To identify the protein targets of sidA, the Laub lab performed a suppressor screen for mutations allowing cells to form colonies despite sidA overproduction. By directed sequencing of candidate genes, most of these mutations were mapped. However, one suppressor strain did not contain mutations in any of the known cell division genes and was directly sequenced to find the mutation.
In order to generate libraries from this strain, the BioMicro Center piloted a new protocol using the Nextera tagmentation system. Standard approaches using fragmentation had appeared to be unsuccessful, possibly due to the high GC percentage of Caulobacter. The tagmentation system uses a Tn5 transposase to insert sequence tags into intact genomes, both tagging them and fragmenting them at the same time. This reduces the number of operator steps and avoids the need for sonication. With the suppressor samples, the Nextera system was directly compared to sonicated DNA prepared with the SPRI-TE. The sequencing data showed that the Nextera system was able to produce very even and consistent coverage of the genome and is now being offered as a service through the BioMicro Center.
Screening yeast libraries for genes involved in DNA damage response - SAMSON LAB - Biology-BE-KI-CEHS
A myriad of new chemicals have been introduced into our environment and exposure to these agents can have detrimental effects on biological systems. Many of these chemicals are thought to have mutagenic activity. Analysis of the cellular response to these potential toxins using S. cerevisiae can provide a description of systems level responses to environmental stress and identify new pathways of DNA damage response. High-throughput techniques have become important tools to establish and clarify toxicity-modulating pathways of potential environmental carcinogens.
The BioMicro Center has worked closely with Laia Quiros-Pesudo from the Samson lab in multiple complimentary screening methods to identify the cellular systems that respond to DNA damage. The initial screening method involved performing barcode-sequencing (Bar-Seq) described by Smith et al. (Genome Res. 2009. 19: 1836-1842) on a haploid yeast knockout library. In Bar-Seq, each knockout strain is identified by two unique barcode sequences ("uptag" and "downtag" barcodes) that can be amplified from its genome and identified using Illumina sequencing, allowing the whole library to be grown together in a single vessel. The importance of each knocked out gene is then measured by comparing the frequency of the strain in the initial knockdown library pool to the frequency after subjecting the pool to an environmental stress which is summarized as the fitness defect ratio. Multiple conditions can be tested simultaneously by using a second molecular barcode added to the library that identifies the experiment.
Using the Bar-Seq approach, the Samson lab was able to simultaneously analyze the frequencies of ~4,800 strains of S. cerevisiae in up to 19 treatments and doses in a single Illumina sequencing lane. The Bar-Seq method was able to reproduce previous results using a solid agar assay and the alkylating agent MMS (Begley et al, 2004). In addition, new groups of sensitive strains have been identified and analysis of these new pathways is currently underway.
In addition to Bar-Seq approaches, the Samson lab is directly screening GFP-fusion libraries to identify proteins that respond to environmental stress. GFP-fusion libraries monitor both changes in protein expression and localization as a result of chemical exposure instead of survival. In these experiments, each strain is individually screened across a number of conditions, requiring significant automation to make the experiment feasible.
In order to perform these screens, the Samson lab has relied on the Tecan EVO 150 liquid handler in the BioMicro Center and the Cellomics ArrayScan VTI HCS reader available through the CEHS Genomics and Imaging Core (similar instruments are also available through the Whitehead Institute and will be available through the Koch Institute). The Tecan EVO150 performed the cellular treatment, fixation and staining of multiple GFP tagged library plates simultaneously and significantly improved the throughput of the library screen. Initial results have been promising.
Processing Very Long Illumina Reads - CHISHOLM LAB - Biology
Different high-throughput sequencing platforms are currently available, and trade-offs currently exist between the cost per sequencing read, the number of reads, and the average read length. The Chisholm lab has been interested in optimizing the Illumina platform for the de novo sequencing of microorganisms. To this end, the Chisholm lab has worked with the BioMicro Center to develop a pipeline that significantly increases the read length yielded by the Illumina sequencing technology, generating sequencing reads that can exceed 250 nucleotides in length. Combined with Illumina's low cost and high-throughput, the procedure expands the range of applications that can be performed with this platform.
Illumina reads tend to decrease in quality with length due to slight errors in incorporation and extension of the growing sequence. To improve the error rate at long read lengths, the Chisholm lab developed an algorithm SHERA (SHortread Error-Reducing Aligner) which uses overlapping paired-end reads to create long and accurate composite reads. SHERA allows more than 87% of the paired-end sequencing reads to produce longer composite sequences with less than 1% of paired reads incorrectly aligned. The quality score of each overlapped base is re-evaluated to take into account the information from the two paired-end reads. The Chisholm lab sequenced a marine metagenomic DNA sample using 454-FLX and the Illumina paired-end overlapping procedure, and found that the taxonomic classification results are highly platform-independent, demonstrating that that composite sequencing reads constitute a cost-effective alternative to pyrosequencing.
The creation of high-quality very long Illumina reads is not only applicable to metagenomics sequencing. The BioMicro Center is currently working to deploy this algorithm for many other applications including amplicon sequencing, transcriptomics, de novo assembly and resequencing for mutation detection. We anticipate a strong growth in very long reads in FY2011. This work has been accepted for publication in PLoS One.
High Throughput Metagenomics Using Illumina Sequencing - ALM LAB - BE - CEHS
Understanding the role of the human microbiome in health and disease is an emerging field, and has been targeted as a major NIH Roadmap Initiative. Microbial community analysis by 16S rRNA sequencing is a key component of microbiome studies, together with whole genome sequencing and metagenomics. The BioMicro Center has worked with the Alm lab to establish and optimize an experimental approach to generating partial 16S rRNA sequences that is orders of magnitude less expensive than conventional methods, thus enabling unprecedented resolution in microbiome comparisons using the Illumina Genome Analyzers.
The use of Illumina sequencing for assay microbiomes has been limited by read length and by financial considerations. In addition, homopolymeric sequences are very difficult to process using the standard Illumina image analysis software. Improvements to read length introduced by Illumina and careful selection of priming sites have addressed the former issue. The expense of the reads has been solved by using very highly multiplexed lanes. While each lane of Illumina sequence costs $3,300 for the read lengths needed for this project, multiplexing the samples (up to 96x) dramatically lowers the cost per sample. Barcoding the sample also allowed the Alm lab to bypass the lack of complexity in ribosomal reads as the highly diverse sequences meet the criteria needed for spot finding. Data from this project has been used in an NIH grant application and in a patent application.
Testing of NuGen's SPIA Technology for Illumina Sequencing - BURGE LAB - BE - Biology - KI
One of the key limitations of RNA-sequencing is the relatively large amounts of RNA required for each sample. While recent protocols have reduced the amount of total RNA input down below 1g, the initial protocols required 5-10g of material. This is several orders of magnitude higher then we routinely use for microarray analysis.
While most microarray labeling protocols are inappropriate for Illumina sequencing in that they use cRNA as their labeled materials, the NuGen kits used by the BioMicro Center since 2009 are unique in that they use amplified cDNA which has several benefits to microarray analysis. We were particularly interested in the ability of NuGen to handle amounts of RNA in the sub-nanomolar range which could allow next-generation sequencing of RNA from single cells. In order to test the viability of this approach, we established a collaboration with NuGen and with Dr. Chris Burge of the Biology and Biological Engineering departments.
To establish the robustness of the NuGen system for RNA-seq, two mRNA samples were isolated from control and UPF-1 knockdown cells and were prepared either with the NuGen kit (by NuGen technicians) or with standard RNA-seq methods from Illumina (prepared in the Burge lab). The NuGen samples were prepared across a variety of concentrations, and seven paired-end libraries were run. Differential error rates, coverage, sensitivity and differential expression were all calculated by the Burge Lab.
Our results demonstrated that the NuGen kit, unfortunately, has a number of issues that are concerning in the RNA-seq environment. Analysis of coverage showed very uneven coverage of exons, likely due to the semi-random-nonamers that NuGen uses in preparing the cDNA. In addition, the level of noise introduced in differential expression was quite large, at least for an experiment with subtle changes in expression. Our results have discouraged us from focusing on NuGen protocols for looking at RNA-seq data. It has even raised questions to us about the quality of the NuGen kit for exon array analysis, though other whole transcriptome amplifications are probably no better.


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subset: commoncrawl


text:
Cost of Attendance (COA) is the estimated amount it will cost to attend Philadelphia College of Osteopathic Medicine. Figures include tuition, fees, books, supplies and reasonable costs for living expenses.REQUEST PROGRAM INFORMATION
|Tuition and Fees1
|Contact the PCOM School of Professional and Applied Psychology for program course sequence information
|Cost per credit
|$1,226 per credit
|School Fee
|$188 per term
|Books, Supplies, and Equipment
|Books & Supplies
|$433 per term
|Living Expenses2
|Living Expenses
|$7,131 per term
1 Students are charged tuition and school fees each term. Tuition rates generally increase by three percent (3%) each year. If students are paying any portion of their tuition out-of-pocket, they must make payment directly to the PCOM Bursar Office by each term's tuition due dates.2 Living expenses include reasonable costs for room (rent, utilities), board (food, groceries, necessities), school-related transportation (car insurance, gasoline, public transit), health insurance, personal expenses and student loan fees.When student loan and scholarship payments disburse to PCOM, they first pay toward a student's tuition and fee charges. Students then receive any remaining funds as "refunds" to help them meet their personal living expenses. View Disbursements and Refunds for more detailed information.


text:
As the parent/guardian of a student who receives exceptional education services in the Troup County School System, we wanted to inform you about your options regarding public and private school choice. Under the state law passed by the Georgia State Legislature in 2007, parents/guardians of students who receive exceptional education service may choose to transfer their child to another public school or private school in Georgia.
Letter to Parents


text:
codingdimes needed a new logo design and created a contest on 99designs.
A winner was selected from 257 designs submitted by 51 freelance designers.
Need a fresh, creative, Web 2.0 logo for our startup website
We are looking for a 2-3 color logo (blue,orange?) with an optional accompanying symbol. Gradients are okay, depending on how its used. I'll provide feedback.
Logo must contain CodingDimes.com. The '.com' can be subordinate.
I'd like the design to include a graphic element which can beused as a favicon.
CodingDimes.com is a consulting startup specializing in application development and training. The 'Dimes' come from a basketball term meaning 'assist'. Although I could talk basketball (preferably college basketball) all day and night, the logo has nothing to do with basketball or money.
Be creative but keep it simple!
Update: One concept that I think is worth mentioning is related to 'Dimes' (see urban definition above) in the brand name. Its slang for the word 'Assist', meaning to help someone get something done. A logo that represents something of this nature would be nice. However, its not mandatory.
- 2-3 color maximum. I prefer to have a shade of blue in the design.
- prefer horizontal design with logo to the left.
- prefer to have .com under the 'es' in CodingDimes
- logo on a white background in AI vector format
- logo on a black background in AI vector format
- shadows, gradients, reflections, etc.
- anything cartoon-ish
- complexity
- unprofessional
Find out how a design contest works or learn more about our logo design services.


text:
Tuesday, Oct. 17, 2006
|Meeting and Background Information
The Part B Final Regulations implementing the 2004 reauthorization of the Individuals with Disabilities Education Act (IDEA) were published in the Federal Register on Aug. 14, 2006, and can be accessed on the Department's Web site at www.ed.gov/IDEA.
To provide the public with an overview of the regulations, the Office of Special Education and Rehabilitative Services (OSERS) will be hosting a series of community-based public meetings. These public meetings will serve two major purposes. First, the meetings will provide the public with an opportunity to learn about the major concepts and principle changes in the new regulations. Second, the meetings will serve as a mechanism for the public to learn about and obtain some of the many resources and supports available from OSERS to assist in the implementation of these regulations.
|Tuesday, Oct. 17, 2006
Reception: 5:30 p.m.6:30 p.m.
Meeting: 6:30 p.m.9:00 p.m.
The Conference Center at
Penn State Great Valley
Each meeting will begin with a reception at which OSERS Assistant Secretary John H. Hager and Office of Special Education Programs Director Alexa Posny will be available to meet with the attendees on an informal basis. The reception will be followed by a presentation about the regulations, which will include a taped welcome from Secretary Margaret Spellings, information about the regulations and the IDEA Web site, as well as an opportunity to ask questions about the regulations and OSERS' implementation plans.
There is no advanced registration for these public meetings, nor any cost. Registration will take place at the door. Interpreters will be available for individuals with hearing impairments.
MS Word (215K) | PDF (170K)
|Tanya Kosinski, 202-245-7407, email@example.com


text: Constructing Oil, Gas Lease Agreement December 8, 2012 05:17 PM Tweet It's a good idea to visit a drilling site and a working well to get a perspective of the project and the implications for your land. © Joyce Meyer TAGS: Marketing, Overseas December 18, 2014 By Farm Journal Editors Email Tips for constructing an oil or gas lease agreement You've heard the gossip at the grocery store. You've seen the headlines in your local newspaper. The oil and gas companies are coming to town. Signing up for a lease seems like an easy way to boost your bottom line, but at what cost to you and the future of your farm? Here are some things to think about before you sign on the dotted line. Education. Before signing a lease, take some time to visit a couple of well sites for a visual perspective of their size and scope, encourages Eric Johnson, an attorney and partner with Johnson & Johnson Law Offices, based in Canfield, Ohio. "You need to have a picture in your mind of what these sites look like and how big they are," Johnson contends. He says farmers can expect, in a best-case scenario, a minimum of 10 acres to be taken out of commission if a well is placed on their property. To boost their knowledge about the oil and natural gas industry, Johnson also advises farmers to go online to read articles and check out other land owners' experiences on YouTube. Know the language. "Probably the biggest mistake I see is not understanding the lease," notes attorney Ronald Coyer of S.R. Law in Slippery Rock, Pa. "I always try to go through an education process with clients and walk them through the entire lease line by line," Coyer says. "Here's what the lease means, and here's how it affects what you're doing—because it can have some effects." Coyer says to carefully review any protective terms. Ask yourself: What rights does the oil or gas company have? What can you as a landowner do? How does this affect my land? How does it affect the resale value? Can I keep the oil and gas rights if I decide to sell the land? Cost and no-cost leases can be particularly confusing to muddle through. A cost lease means the oil or gas company can charge you a certain percentage on your royalties for production costs. "A lot of these leases are written in a manner that if you're not used to understanding and reading them, you could read it and believe they can't charge you anything," Coyer says. He says another item to watch for is the Pugh Clause, which states that once a land lease's primary term expires, the oil or gas company must release any acreage that isn't in production. "All of these companies look to the effect of, how do we hold onto this piece of property without really doing something because we want to maybe keep it in our bank of assets and pull it out when we need it," Coyer says. For example, if you sign a lease with no drilling depth provisions, the company can drill a shallow well, keep it in production cheaply for as long as it wants, and later go in and deep-drill. "That upsets landowners," Coyer says. "We see a number of those every week. We're seeing them a little less now because people are becoming more educated." Basically, Coyer says, you need to know what addendums (or amendments) are and how they work. You can add addendums to a contract that overrule what the original contract states. This is where you can build in many protections and provisions. Know when to sign. How do you get the best price and protective terms? "Obviously, you want to sit down with a lawyer who has experience with this process and have him review the lease," Johnson says. His typical service charge is $500, a small investment that could provide thousands of dollars in return. "A lawyer knowledgeable about these types of leases should be able to provide some good guidance on the financial terms, and even help make sure you don't leave money on the table," Johnson notes. Coyer says that timing and location are pivotal to getting the most bang for the buck with land leases. "People have to understand that it's about location and probably a bigger aspect, timing, in that market," Coyer explains. "It's like anything else. If there's no demand for something, the price goes down. If there are multiple players and a high demand, the price keeps moving." When oil or gas is first discovered in an area, companies work the area to lease as much land as they can, regardless of whether they will actually drill there—what Coyer calls a "land grab arrangement." As more and more land is leased under these blanket arrangements, eventually competition between companies starts to dwindle, and the major players emerge depending on who leased the most land. The land grab subsequently slows, and those major companies start building and developing their drilling units. "People often ask me, 'When's the right time?'" Coyer says. "It's hard to know, but when we get toward the end of the blanket lease arrangement period, before they start unit leasing, that's probably the time when you can convince an oil and gas company to give you the best protective terms with the most addendums and probably the highest value. "It's just like anything else. The more entities that are trying to get the same thing, the prices escalate; when they get closer to drilling, the prices and terms, along with the protections that go with it, start being pulled back off the table." In a best-case scenario, experts say farmers can expect a minimum of 10 acres to be taken out of commission when a well is established on their property. That's important to factor into the overall ROI equation. Don't be intimidated. While there are many horror stories of landowners being railroaded by a company or blindsided by terms in their lease, Coyer says that most reputable oil and gas companies are willing to work with the landowner to come to a mutually beneficial agreement. "Most of the land agencies that are coming in now are saying, 'This is what we'd like to do. What else can we do for you to make this work?'" he says. Johnson agrees with Coyer's assessment. "Most of these companies are staffed by good, hard-working people," Johnson says. "A minority may try to cut corners, but most of them don't want their reputations sullied—that's not in their best interest." Coyer adds that many companies are willing to help landowners make improvements to roadways or farmland, and do their best to keep the land in good condition. However, he also cautions that there are some companies who still conduct poor business, trying to strong-arm the landowners who sign leases with them. For more information on the issues and faces behind the energy explosion in rural America, including photos and videos, visit www.FarmJournal.com/energy_boom You can e-mail Katie Humphreys at firstname.lastname@example.com. Back to news


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Whether you are looking to buy or sell a house, town home, condo, land, farm, or commercial property in or around Russell County, including the towns of Phenix City, Ladonia, Smiths Station, Columbus, Fort Benning, or the surrounding areas then you have come to the right website. Here you will find a wide array of information about local real estate for sale.
Please click the Search Alabama Homes or Columbus Area Listings link to find your next dream home.
By working with a local Realtor you have the advantage of being able to access all local listings available through the Multiple Listing Service, MLS. Using a Realtor also gives you the comfort of knowing that you are working with the most qualified professional who is dedicated to you as the buyer or seller and will always be looking out for your best interests and striving to make the process of buying or selling real estate as comfortable and pleasant as possible.
Please click the Contact Us link or use the phone number to contact your Realtor today!
© Systems Engineering, Inc.


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BEDFORD, Mass. — The Aero Club of New England will host its second annual Reese Dill aviation safety lecture on Thursday, Nov. 7, at the USS Constitution Museum in Charlestown, Mass. The event will feature Capt. Greg Wooldridge, a three-time commanding officer and flight leader for the U.S. Navy's Blue Angels, as its keynote speaker.
Wooldridge will share his personal experiences as a member of the famed Blue Angels and their commitment to flight safety. Capt. Wooldridge has enjoyed a remarkable career in naval aviation, serving for 27 years in the U.S. Navy.
"We are honored to have such a distinguished guest as our keynote speaker," said Paul S. Diette, the Aero Club of New England's Safety Programs Co-Chair. "Here is an opportunity to get the inside story of what it takes to be a member of the Blue Angels, the Navy's flight demonstration squadron, and what these men and woman do to minimize the risks of close formation aerobatic flying. No doubt there will be lessons learned that are applicable to any pilot and all forms of flying."
This is the Aero Club of New England's second annual Reese Dill safety lectureship event, which is named in honor of Reese Dill who was a skilled aviator, inventor, long-time member, and past president of the club.
The event is open to the public for a $35 fee and advanced reservations are required.
For more information: ACONE.org or call Paul S. Diette at: 508-339-1520.


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Safety Management
The aim of the Safety Management System (SMS) is to ensure airside safety at Hannover Airport. Preventing accidents in order to protect people and property is a matter of top priority.
To achieve this objective, the already high safety standards are checked and further improved on a permanent basis.
Partners and customers of Hannover Airport are welcome to use our Safety Management Portal which offers up-to-date information and allows you to submit safety reports.
The following user data are needed in order to access the system and submit a report:
User name: SMSUser
Password: sms_haj


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text:
codegorilla needed a new logo design and created a contest on 99designs.
A winner was selected from 44 designs submitted by 10 freelance designers.
I am the new State Chair for Texas. I am changing everything from top to bottom. I have competed in this sport for over 15 years and competed locally, nationally, and internationally. I have a background in sports photography, and art.
My target audience includes the members of the USA Powerlifting organization in the state of Texas, potential members, and powerlifting enthusiasts in general. We have members of all ages ranging from 14 through 80 years old. We have teams that represent high schools, colleges, and the gyms the atheltes train at.
- needs to be an ICONIC logo that we can use to brand everything, from clothing to stationary.
- USA Powerlifting (USAPL) is the main organization but I would like to see Texas as the dominant wording.
- state colors are red, white, and blue.
- iconic symbols for Texas include the shape of the state, the flag, lone star, branding irons, etc.
- BOLD! Remember everything is bigger in Texas :)
- Classic
I would NOT like to see silhouettes of muscle men lifting, longhorns or cowboys!!
For more information on the USA Powerlifting organization check out the following links below:
Find out how a design contest works or learn more about our logo design services.


text:
black bart wrote:Dear Aunty,
I has a friend what fancies his cabin boy but he be too coy like to approach the subject directly. This friend of mine also as a penchant for wearing a oomans undies beneath his Pirate Attire. If the men should ever find out this friend of mine would likely get marooned on Canvey Island.
What should I do dear Aunty?
Arrr! Oi reckons ye...Oi means ye're friend...should just come out o' Davy Jones' closet. Oi've got a friend who 'as done just that, and 'e be finding that most o' his ship-mates be the same as 'im.
And FSM be likin' flimsy morals o' all sorts. :fsm_touching:


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AP Environmental Science
Exam Practice
Exam Day 2019
May 06
12:00 PMView AP Exam calendar.
You can use the resources below as you prepare for the AP Exam.
Click here for details about the exam format.
Sample Questions
You'll find sample multiple-choice and free-response questions in the AP Environmental Science Course Description (.pdf/529KB).
Sample Responses
Student responses to past exam free-response questions are available on AP Central.
Exam Practice Tips
Check out our Exam Practice Tips.
Free Response Questions and Scoring Guidelines
Free Response Questions and Scoring Guidelines: login
The following resources contain the remaining available Free Response Questions and Scoring Guidelines for the AP Environmental Science Exam. To access the files below, you need to log into your College Board account. If you do not have a College Board account, you can create one by selecting 'Sign In' in the header and following the prompts to Sign-Up.


text:
A mysterious new Watch Dogs website has appeared online.
The site, called 'WeareData', asks users whether they have "ever experienced the power of data", and tells them to "be prepared" for Watch Dogs WeareData.
The keywords 'Data', 'Big Brother', 'Network', 'Technology', 'Experience', 'City', 'Release', 'Hack' and 'Transportation' also appear in the site's metadata.
But what is it? Ubisoft has yet to release details on WeareData, but it could be linked to the game's companion app, set to act as a multiplayer tool allowing mobile users to hack into helicopters and road blockers to help other players.
Watch Dogs releases this November on Xbox One, Xbox 360, PS4, PS3, Wii U and PC.
Source: wearedata.watchdogs.com


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No (more) baby mother drama?
A few weeks ago there were reports online suggesting that Gael Garcia Bernal found out that his baby mother Dolores Fonzi had been getting it on the side and that maybe the kid, a son called Lazaro, wasn't his so he was demanding a DNA test to determine paternity. GGB immediately instructed his people to tell People that none of it was true. He and D are apparently still together, happy, and there is no fatherhood question mark looming over little L's head. It was a curious non-scandal though. And while the parenthood issue was resolved, there remains the infidelity charge. The denial came without any mention of lawsuits. Not that that means anything. But it's all very soap opera, non?
Here they are, the happy family, in Madrid today. Well, GGB looks happy. Dolores, photo assumption, looks miserable. That's obviously a photo assumption with a very "me" slant given that that's totally the face I would have if I ever had a baby.
Photos from Bauergriffinonline.com


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6:30 am
WordWorld "Bed Bugs/Snug As A Bug" Bed Bugs - When the bugs accidentally break Frog's brand new bed, they scramble to put it back together and make things right. Just one problem. They can't tell the letters 'b' and 'd' apart! Can they figure out which letter is which - and rebuild Frog's bed - before Frog returns home? Snug as a Bug - Frog helps insomniac Bug figure out the end of the rhyme "snug as a bug in a . D
7:00 am
Sesame Street "Don't Wake The Baby" Leela hums as she rocks her sleeping baby niece, Nila, in a bassinet in the arbor. Telly and Elmo come bounding in loudly trying to offer their help, but Leela insists that she doesn't need any assistance. She's hoping Nila will stay asleep as long as there aren't any loud noises on Sesame Street. D
8:00 am
Curious George "George and Allie's Lawn Service/Curious George's Scavenger Hunt" George and Allie's Lawn Service - George and Allie are crazy about Mr. Renkins' new kids -- baby goats named Suzy and Sammy. But when the goats keep escaping their pen and chewing on everything in sight, can George and Allie find a way to keep their new friends penned up? Curious George's Scavenger Hunt - It's a race around the city as George, Marco, and Chef Pisghetti go on a shape-finding scavenger hunt to raise money for the animal shelter. They find rectangles, stars, and circles, but how are they ever going to find 100 diamonds?. D
8:30 am
Cat in the Hat Knows A Lot About That! "Top of the Sky/Jiggle Bones" Top of the Sky: Nick, Sally and Cat are playing with their toy rocket. Using the Cat's "Whoa-Look-At-It-Go-Launcher," he shoots the rocket up up and up! But now they need to go find it. Off to the top of the sky! They pass the treetops, and the mountains. D
9:00 am
Super Why! "The Three Billy Goats Gruff" Gramma will not let Red into the kitchen to get some extra special Red Velvet Cake - and it's making Red extremely mad! The Super Readers head into the story of the Three Billy Goats Gruff, who really want to cross the Trolls bridge#only this time the Troll gets to tell his side of the story. The Super Readers wonder if anyone can make those rude Billy Goats Gruff behave, and Red figures out what Gramma needs to hear. Educational Objectives: To learn the importance of good behavior, discover the alphabet and identify the letters P-O-L-E. D
9:30 am
Dinosaur Train "Dinosaur Camouflage/Family Scavenger Hunt" Dinosaur Camouflage - Buddy and Tiny meet a shy dinosaur, Leslie Lesothosaurus, who is very good at hiding. The kids end up persuading Leslie to stop hiding, and come out to play with them. Scavenger Hunt - The Pteranodon family travels to the Big Pond for a Scavenger Hunt. D
10:00 am
The Electric Company "Madame President" Lisa and Francine face off in the election for Book Club President. Can Lisa's campaign message win out over Francine's dirty politics?. D
10:30 am
Wild Kratts "Birds of a Feather" When Aviva reveals that dancing is just not her thing, Chris and Martin set out in the New Guinea forest to inspire her to give dancing a try by uncovering some of the greatest dancers in the creature world - the Birds of Paradise. They soon find that the male birds use their showy bright feathers and extraordinary dances to display to potential female mates that they are healthy and strong. But when the gang gets caught up in Donita's plan to capture these fashionable birds for her latest clothing line, Aviva must put her dancing skills on the line to save them. D
11:00 am
WordWorld "Bed Bugs/Snug As A Bug" Bed Bugs - When the bugs accidentally break Frog's brand new bed, they scramble to put it back together and make things right. Just one problem. They can't tell the letters 'b' and 'd' apart! Can they figure out which letter is which - and rebuild Frog's bed - before Frog returns home? Snug as a Bug - Frog helps insomniac Bug figure out the end of the rhyme "snug as a bug in a . D
11:30 am
WordGirl "Meat-Life Crisis/Mobot Knows Best" Meat-Life Crisis - When the Butcher can't summon the meat to defeat WordGirl he decides to go live at home with his dad, Kid Potato. He convinces his son to join him at the ballpark to work a as food vendor. Will this be the end of the Butcher's villainous ways? Mobot Knows Best - It's parent-teacher conference day at Woodview Elementary School. D
12:00 pm
Outdoor Idaho "Climbing Idaho" Today's rock-climbing enthusiasts support climbing gyms, clubs and advocacy groups - no longer the exclusive band of wanderers of just a few decades ago. The heights they achieve throughout the state require skills, levels of expertise, techniques and climbing methods as varied as the state's terrain. Cameras follow as they seek to conquer nature's rock walls.G
12:30 pm
Pati's Mexican Table "Hibiscus Flower" In Mexico, it's not uncommon to find both food and water deliciously flavored with hibiscus flowers. Commonly known as jamaica, they have an intensely herbal, fruity taste. Pati shows viewers where to buy these flowers and then shares recipes such as: Jamaica Water; Spinach Salad with Caramelized Pecans and Jamaica Vinaigrette; Steak Tacos with Jamaica and Jalapeno Syrup; Gourmet Jamaica Popsicles. D
1:00 pm
Katie Brown Workshop "Vegetable Medley" Learn to cook, grow, display and love vegetables with exciting projects and delicious recipes. In the Kitchen: Creamy Cauliflower Soup, Colorful Chopped Root Salad and Quick Vegetable Roll-Ups. Projects: Sculptural vegetable centerpiece. D
1:30 pm
Paint This With Jerry Yarnell "Back Country Boulders, Part 1" In this episode, Jerry will show viewers steps of painting rock in relationship to a back-country waterfall on a textured canvas. Great for basic to advanced painters covering layout, sectional under-painting, gradation of values, and use of chisel-edge brushes. D
2:00 pm
Essential Pepin "Fabulous Fins" Cured Salmon In Molasses; Tuna Steaks With Tapenade Coating; Tuna Tartare On Marinated Cucumbers; Poached Salmon In Ravigote Sauce; Ray Meuniere With Mushrooms. D
2:30 pm
On Story "The Humor and Heart of Animation" The writers behind the beloved animated classics KUNG FU PANDA 1 & 2, "King of the Hill," and THE IRON GIANT, Jonathan Aibel & Glenn Berger and Tim McCanlies discuss the comedy, action, and emotion that go into creating the worlds and characters of the beloved and popular films that appeal to audiences of all ages. And then enter the magical world of the short film Dreamgiver by director/producer Tyler Carter. What fanciful adventures happen after we go to sleep?. D
3:00 pm
Dialogue "Jac Venza" Host Marcia Franklin talks with the founder of several long-running PBS arts series - including GREAT PERFORMANCES and LIVE FROM LINCOLN CENTER. He and Franklin talk about his long career in public broadcasting, the potential effects of cuts in national grants programs, and the trends he is following in the arts.G
3:30 pm
MotorWeek "Mazda6" Road Test: 2014 Mazda6. Road Test: 2013 Land Rover Range Rover. Goss' Garage: Fork Maintenance. D
4:00 pm
America's Test Kitchen from Cook's Illustrated "Asian Takeout Favorites" Test cook Julia Collin Davison shows host Christopher Kimball how to make the ultimate Sichuan Stir-Fried Pork in Garlic Sauce. Then, equipment expert Adam Ried reveals his pick among blenders in the Equipment Corner. And finally, test cook Bridget Lancaster uncovers the secrets to making perfect Thai-Style Stir-Fried Noodles with Chicken and Broccolini right at home. D
4:30 pm
Crafting at the Spotted Canary "Things That Adorn Everyday Life" Wear your creativity as we adorn our everyday life with hand made accessories. Learn how to add those decorative accents that define your personality. The first segment shows how to make a decorative cuff - it's more fun than a regular bracelet and has a texture that makes it interesting to wear. D
5:00 pm
Globe Trekker "Around The World - Pacific Journeys: Tonga to New Caledonia" Zay Harding concludes his grand Pacific adventure heading further west across the South Sea Islands into Tonga, Fiji and New Caledonia - islands whose histories have been shaped by brutal tribal conflicts and British and French colonization.G
6:00 pm
The Artist Toolbox "John Legend" In this episode of The Artist Toolbox, get a taste of the performance chops of six-time Grammy Award-winning recording artist John Legend with host John Jacobsen. More than just good looks and what Legend has called a 'presumptuous' name, Legend's innate musical ear and talent for fusing his early gospel influences with hip-hop, jazz and even classical music, has found him working with top names such as Lauryn Hill, Alicia Keys, Janet Jackson, Talib Kweli, Jay-Z, Britney Spears, Eve, Common, the Black Eyed Peas and longtime producer and friend, Kanye West. So adept at weaving myriad sounds and genres, Legend seems to have created a signature sound, if not genre, of his own. D
6:30 pm
Sun Studio Sessions "Delta Blues Experience, Part One" Delta Blues Experience Part 1: In this episode, Delta blues musicians; including Bobby Rush, James "Super Chikan" Johnson and Cedric Burnside perform blues classics and originals while sharing stories about their songs, the stories behind them and the historical importance of blues music. D
7:00 pm
Austin City Limits "The Shins/Dr. Dog" This show highlights two alternative rock groups. The Shins play from their latest recording Port of Morrow while Dr. Dog performs selections from Be the Void.G
8:00 pm
Great Performances at the Met "Un Ballo In Maschera" Marcelo Allvarez, Dmitri Hvorostovsky and Sondra Radvanovsky star in David Alden's production of Verdi's opera. D
10:30 pm
Sun Studio Sessions "Delta Blues Experience, Part One" Delta Blues Experience Part 1: In this episode, Delta blues musicians; including Bobby Rush, James "Super Chikan" Johnson and Cedric Burnside perform blues classics and originals while sharing stories about their songs, the stories behind them and the historical importance of blues music. D
11:00 pm
Austin City Limits "The Shins/Dr. Dog" This show highlights two alternative rock groups. The Shins play from their latest recording Port of Morrow while Dr. Dog performs selections from Be the Void.G


text: The Hit Crew (Music Downloads Not Rated by the ESRB) Join the movement and get the best dance music. Expand your track list to keep the dance party going! Download a free demo of "Toxic" - The Hit Crew. Written by Cathy Dennis, Christian Karlsson, Henrik Jonback and Pontus Winnberg. Published by BMG Gold Songs (ASCAP) o/b/o Murlyn Songs AB (ASCAP) administered by BMG Rights Management (US) LLC / Colgems All rights reserved. Used by permission. Courtesy of Turn Up The Music Licensing. By arrangement with The Orchard.


text:
- Lost your password? Create a new password of your choice.
- No password? Create a free account and download a free eBook.
BAS Travel/Study Program
Visit Biblical archaeology sites, discover ancient Bible history, and hear top archaeologists and Bible scholars share their latest research and findings.
Browse Topics
- Ancient Cultures
- Archaeology Today
- Biblical Artifacts
- Biblical Sites & Places
- Biblical Topics
- People & Cultures in the Bible
The Rock-Hewn Churches of Lalibela
Megan Sauter
With 11 rock-hewn churches, Lalibela, Ethiopia, is understandably a place of pilgrimage for those in the Ethiopian Orthodox Church. Explore Lalibela's spectacular subterranean churches in this web-exclusive slideshow.


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Please consider employing a Student Intern from either Staples or Weston High Schools. Both schools have created programs for their Senior Students, (approximately for 4 weeks, beginning the middle of May through the middle of June), for the opportunity to work for a local business of interest. This is a great way to connect with our community, possibly groom a student for future employment and enlighten yourself about the current generation that will be entering the work force in four short years!
For Staples High School the contact person is
Michele Howard & Denise Pearl
For Weston High School the contact person is
Meredith Starzyk


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Friday, April 30, 2010
Photo: Tour of Romandie - Stage 1 sprint finish
1 comment:
- Anonymous03 May, 2010 13:02
Cool picture, but you should give credit to these photographers whose works you borrow. I'm not saying that you should have to pay them royalties or anything, because you're just a blog, but at least give them the credit their artistic talent deserves. Eh?ReplyDelete
And who is the Garmin rider on the left? I don't know why, but he looks like a lazy puppy dog sprinting across the line...just cruising along on a warm Saturday afternoon. Haha.
If that was a mountain stage you could have a competition to ask how many of the riders in the photo would still be in the sport after a year (ie, who was going to have to stop because of their doping)?
Pappillon welcomes your comments and encourages your participation. However, in commenting, you agree that you will not:1) Post material that infringes on the rights of any third party, including intellectual property, privacy or publicity rights. 2) Post material that is unlawful, obscene, defamatory, threatening, harassing, abusive, slanderous, hateful, or embarrassing to any other person or entity as determined by Pappillon in its sole discretion. 3) Impersonate another person.


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Heading into Thursday night's game, there were just three teams in the NFL who were among the league's top 10 in both offense and defense. Just three teams that were among the league's statistically proficient on both sides of the ball.
The Cowboys (3-5), the Lions (4-4) and the Broncos (5-3) are the three. But what was once considered a sign of balance for a playoff hopeful is now an uncertain gauge at times.
Dallas has not looked like a playoff team at any point since the Cowboys' opening-week win over the Giants. Detroit has beaten just one team that currently has a winning record — Seattle, at 5-4.
And the Broncos have steadily climbed from 2-3 to their current spot at the top of the wobbly AFC West with a three-game winning streak.
Which brings us to the reason a cliché becomes a cliché — because it comes up over and over again.
And turnovers are the NFL's cliché, with coach after coach, player after player, week after week, season after season talking about the importance of "the turnover battle." Commence eye-rolling if you need to, but it's been true before and it's true again.
It's also the reason the Broncos, despite all of the good they have done in winning three of their last four games, cannot yet be considered at the front of the line in the playoff chase. They are considered to be a potentially tough out in the postseason because of what Peyton Manning brings to the table. But until they get their turnover issues in order, they will not be all they can be.
At the moment, the Broncos are 23rd in the league in turnover margin at minus-4, a total boosted significantly by 10 lost fumbles in eight games. The 10 teams at minus-4 or worse so far are a combined 33-49 (.402).
Among those 10 teams, the Broncos and the Colts (5-3) are the only teams with a winning record.
The league's top 10 teams in turnover margin before Thursday night's game kicked off to open a new NFL week are a combined 55-25 (.688).
Seven of the eight division leaders at the moment are among top 10 in the category, ranging from plus-16 to plus-3. The only division leader not among the best in turnover margin is the AFC West-leading Broncos.
The Bears (7-1) lead the league with a staggering 28 forced turnovers. The Bears are tied for the league lead in fumble recoveries — with New England at 11 — and are tied for the league lead in interceptions, with the Giants.
But the Falcons (8-0) are also fourth in turnover margin, at plus-10. The Texans (7-1) are sixth at plus-8.
"That's one thing when you look at it, that always has an effect on what you're doing," said Broncos coach John Fox. "We'd like to get those things cleaned up. We've done better as we've gone along and that's your goal to keep improving and we talk about those things all the time. We'll keep working on those things."
It is a must for the Broncos because any continued improvement on offense, any improved play on defense will be lost in the bottom line of turnovers.
At least that's what the numbers always say.
Jeff Legwold: email@example.com or twitter.com/jeff_legwold


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Advertising Isn't A Mobile Strategy
from the content-comes-first dept
ReadWriteWeb has an article claiming that the way to beat Google is by having a better ad platform (via Matt Asay). It says that "the company that can corner the mobile web ad market is going to be able to go toe-to-toe with Google." This is getting things completely backwards. Google doesn't dominate the search engine market because it has the best advertising platform. It dominates the advertising market because it has the best search platform. Other companies, such as AOL, that have seen themselves as an advertising company first and a content/applications company second have gone nowhere. This is especially true because the best advertising platforms are tightly integrated with their associated applications. One of the reasons Google's search advertising is so lucrative is that Google figured out how to make its ads highly relevant to users' search terms. This had two benefits. It increased click-through rates, obviously, but more importantly, it made the ads more useful -- and therefore less annoying -- to users. The same principle applies to the mobile space. Mobile advertising will only work if companies figure out how to make the overall user experience positive so that the ads don't scare users away. The way to do that is not to focus on building the best possible mobile ad platform, but rather to build the best possible mobile services, and worry about monetizing them after they've built up a significant user base.
Filed Under: advertising, business models, platforms, strategy


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Nature Calendar, June Description
This June print is 5 x 5 inches. All of Nancy's prints come ready to hang. They are mounted on wood and bound in metal or wood depending on the series you order from. If you are giving as a gift, Nancy's bio is printed on the back of all of her prints. The metal topper that is a part of many of Nancy's prints is hand-painted and all of our printing, painting and assembly are done in Virginia.


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I have been tagged, as they say, to do this "meme" that everyone speaks of. But, well, you'll see.
Four jobs I was really, really bad at:
Bank teller. I was the worst bank teller since the creation of bank tellers. I was fired from the first bank I worked in after one and a half days. Then, in a cruel twist of fate, another bank hired me, and every day I doled out the wrong amount of money to people and chatted everyone up about how I was only doing this for the summer because I went to Wellesley and I was going to be a writer. One day a truck hit the drive-through section of the bank and tore it off, injuring the bank teller who was sitting inside it. Later that day I heard one teller say to another, "Too bad it wasn't college girl in there." That's how assy I was.
Cocktail waitress at the men's bar in a country club. The men's bar. As in no women. I wore a red bow tie, a satin shirt, a short black skirt, fishnet stockings, and a grimace.
Salesperson at Saks Fifth Avenue. I was in the gifts department, which no one ever visited. I stood there and dreamed of sitting. If someone came in and asked if there were any more, say, silver-plated candlesticks in stock, I went into the stockroom, waited a minute, then came out and announced that there were none. I did this not out of malice but rather a frightened ignorance of the workings of the stockroom.
English tutor for intensely charming Japanese businesspeople. One day in early September, one of my tutees told me, "Already I feel autumn flavor completely," and how could I correct that?
Four movies I could watch over and over:
Love and Death
The Ten Commandments
All About Eve
A New Leaf (If you haven't seen it, and I'm sure you haven't, you must.)
Four childhood memories and/or dreams. Guess which is which!
1. I am getting a bath in the sink.
2. I am playing peek-a-boo with my sister and another girl. I'm in my crib, which is covered with a large pink blanket, and my sister and the girl are peeking in at intervals. I am beside myself with joy and terror.
3. King Friday pops up from the space between my bed and the wall and asks me where is his castle, what have I done with it.
4. My entire family goes on vacation and leaves me alone. With a monster. Who's trying to kill me. Geez!
Some celebrities I worshiped when I was little, and names I wanted instead of Alice (also, you know, when I was not an adult)
1. Tony Randall. I think this was a father thing, as my dad is similarly (desperately scrambling to find a word other than "prissy") refined.
2. Cassandra.
3. Sandy Duncan (but why? Was it the glass eye? The Triscuits commercials? I really can't say)
4. Veronica.
Four injuries I have sustained:
1. Concussion. Take one giggly teenager at a beach party, then add a cute boy chasing her around the first floor of the beach house where said party was being held. Put glass door between her and the beach. And voila! Luckily my enormous skull bore the brunt of the impact. And also the door didn't break.
2. Broken tailbone from falling and landing on my knees. Apparently I fell so hard that my tailbone was dislodged, or was rebroken after a alcohol-related ass-landing from years before ( this was extraordinarily painful and necessitated the humiliating purchase and then use of the "ass donut"; avoid this injury at all costs)
3. Broken front tooth (just as that permanent tooth came in! I have always had excellent timing). I tripped on a shoelace and fell on my face.
4. Sprained ankle while at Disney World with my high school chorus. The bad part was that I had to be wheeled around. The good part was that I went straight to the front of the line! Whoever wants to see the Teddy Bear Jamboree, better come with me! The other bad part was that the only person who wanted to see it—including me--was my chorus teacher, who somehow got control of my chair and took me to see it twice.
Four celebrities I have bothered:
1. I was working at a magazine launch party, because I worked for the magazine and didn't have a choice. For some inane reason I am giving out our goody bags at the beginning of the party, such that everyone has to carry around these little plastic bags with our stickers and etc. in them. So Spalding Gray walks in, with a man who looks exactly like him. He says, "Do I have to carry these around at the party?" I think, "OH MY GOD SPALDING GRAY SPOKE TO ME." Then I say, "Why don't I save yours for you." He says, "But won't it get lost? With all these other bags?" I say, "Well, maybe, but you know they're all the same." He says, "I just think you might run out." I say, "Why don't you write your name on the bag" (like I couldn't write his name for him?) and he does; in very careful letters he writes SPALDING across the top. And then he eats all the hors d'oeuvres and goes home.
2. I am on the subway across from some guy I can't stop staring at because he looks so familiar. Did I know him in college? Is he my neighbor? He is trying to read his magazine but is clearly intensely aware that I'm studying him. I am fascinated by how far down he is able to shrink into his jacket. He looks so horrified that I'm beginning to feel insulted and so I stare more aggressively. Something wrong with me looking at you, buddy? I'm not good enough for you? Then I realize it's Steve Buscemi. Ah.
3. Walking down the street, staring at an insanely attractive man who is walking with another man, I run directly into the other man. It is Lou Reed.
4. In college, I meet Gloria Steinem at a friend's birthday party. (My friend has a father who is somewhat fabulous.) All I can say is, "I love you. I mean I really love you and everything you stand for. If I had known you were going to be here I would have read Ms. or something first. Oh my god I'm so drunk. I love you." Actually I have no idea what I said; I was too drunk.


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RIO DE JANEIRO, BRAZIL, March 24, 2009 — Last month, thousands of dancers in full make-up and bright costumes, along with hundreds of elaborate floats accompanied by musicians and rolling PA trucks blasting samba music marched down Rio's famed Sambódromo in celebration of this year's Carnaval. Song after song, samba school after samba school, Shure microphones took charge of vocalizing the passion of the week-long event that marks the beginning of Lent.
Televised globally by Rede Globo, the fourth largest broadcaster in the world, Carnaval is a giant celebration where samba schools, which consist of thousands of performers, compete for bragging rights as the official winner of Carnaval. The schools prepare all year for this one performance where they are meticulously judged on their costumes, choreography, floats, music, harmony, and overall theme. Because music and harmony play such an important role in the outcome, and due to the hectic outdoor RF environment, all of the bands that performed chose Shure wireless microphones to accompany them on their kilometer long, one-hour performance.
Representatives from Shure worked with Pride Music, Shure's Brazilian Distributor, to provide RF coordination and technical support for Rede Globo and Gabisom, Brazil's largest live sound company. They successfully coordinated and monitored the use of more than 78 channels of wireless frequencies using Shure Wireless Workbench (WWB) 5.0 software and UHF-R wireless receivers.
"Our goal was to give full-time technical support to the Gabisom team by managing and monitoring RF using an external scanner that talks to Shure's Wireless Workbench software," said Marinho Alves, Sales Manager for Pride Music. "Because of the constantly changing RF environment, the Wireless Workbench RF scanner and backup frequency features were critical in allowing us to recognize and respond to any RF problems, as the trucks accompanied the performers down the Sambódromo."
In all, 30 UHF-R channels, 12 UHF wireless microphone systems using SM58® and Beta 58A capsules, and 12 PSM®700 personal monitor systems accompanied the schools on their performances without a failure, thanks to daily frequency scans and coordination by Alves and Gabriel Benitez, Training and Education Manager for Shure.
"During one of my Wireless Workbench scans, I noticed two intermittent DTV stations and one of those channels landed right on one of the UHF-R handheld transmitters," said Benitez. "Since this occurred during one of the performances and there weren't any spares available, there was no chance to swap it out or reprogram the transmitter. Fortunately, UHF-R is an extremely agile wireless microphone system, so we didn't experience any dropouts."


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|» News Mentions
|Press Releases | TV and Video
Hotels We Love - The Roxybury Motel in New York
Runaway Now
June 15, 2011
Airfares are through the roof this summer so for someone like me, a short car/ train ride to a get out of New York City is ideal. The Catskills is a perfect weekend retreat and staying at The Roxbury Motel (located in The Catskills) magnifies your vacation into a unique memorable experience.
The Roxbury has recently expanded, doubling their capacity with five new Big King Themes rooms. Each room has been inspired by a 60s, 70s or 80s TV show or film. Who wouldnt want to slumber underneath a steel sculpture canopy of Farrah Fawcetts hair or dance the night away in a Saturday Night Fever-inspired room. Packing The Bee-Gees Greatest hits and a disco dress are optional.
Their other rooms are far from dull. Bring your little black dress, pearls, and long cigarette holder and stay in the Golightly-a-Go-Go Room. Decorated like a Tiffanys box, this room could knock the socks off of Truman Capote. I have to admit my dream room, however, stems from a childhood crush on teen heart throb David Cassidy. Sing along to Come On Get Happy while resting in The Partridge Nest decorated in bright blocks of color. True fact, the bus exterior was inspired by Dutch painter Piet Mondrian. Sounds pretty groovy.
Relax during the day at one of The Shimmer Spas. The Shimmer Spa North is geared more towards couples featuring simultaneous massages and a new Green Steam created from green mosaic tiles from 100% recycled windshield glasses. For those who would like to experience spa treatment alone, head to The Shimmer Spa South for massage rooms, hot tub, sauna, and black-river-rock- foot massaging super shower.
What would a vacation be without cocktails? The Public Lounge is located right across the street. This hip lounge features Flamingo Cosmos, Dorothy Parkas (Manhattans) and Hot Buttered Rum.
So if you are looking to put some flavor into your travel experience, look no further than The Roxbury Motel. Your stay at the Roxbury will be one to write home about.
The Roxbury Motel, 2258 County Highway 41, Roxbury, NY 12474. + (607) 326-7200; www.theroxburymotel.com
Click here to view the story on the Runawaynow.com Website.


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A beautiful, original hand-illustrated greeting card. This card features a floral pattern with the quote "For My Grandmother". A perfect choice to make your grandmother feel special this Mother's Day.
300 gsm premium matte finish paper for maximum longevity.
The high-quality professional paper has a smooth texture with warm white tone and quality rigid feel, creating cards that will last a lifetime.
The card comes in a choice of two sizes:
A6 size (10.5 x 14.8 cm or 4.1 x 5.8 inches) with a brown Kraft envelope.
A5 size (14.8 x 21 cm or 5.8 x 8.3 inches) with a brown Kraft envelope.
The card will be posted by Royal Mail and will reach you within 2-7 business days depending on your location.
We deliver worldwide, and delivery is FREE to the United Kingdom.


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Da......Da Da......Dadadadadadadadada.....
by Cole Smith
5, 2006 - Jaws
Unleashed lets you experience the action from the
perspective of the shark. Its not a bad idea
but if you smell something fishy dont be surprised
if the source of the odor is emanating from your game
definitely have to sit on the middle of the fence
on this one. Jaws Unleashed is neither great nor horrible.
You could say that its both good and bad. Its
kind of like watching a ball game where you favorite
team gets sacked. It may not be a good thing but its
better than not watching the game at all.
we go any further and you dont read to the end
of the review let me get this out of the way now.
If you have any interest in this game at all rent
it. I cant think of any good reason to purchase
it. Having said that, it can be fun to play
but not for long. Ive played Ecco the Dolphin
but Ive never played as a murderous great white.
Its like controlling an underwater biological
30 years after the original movie and the island of
Amity has returned to normal; the shark incident is
nothing more than local legend. But when a new manufacturing
plant sets up production in the region and begins
polluting the waters, the faint strains of the two-note
soundtrack can be heard, foreshadowing a dramatic
role reprisal.
the shark is back and hes pissed (He's called
"Bruce" because that's what Steven Spielberg
named the mechanical shark while filming the original
. Its difficult to tell exactly why hes
pissed but when a 25-foot great white is on a rampage
it really doesnt matter. As Bruce, the gameplay
is relatively open-ended. There are tons of things
to do to fulfill your role as a giant aquatic killing
machine. At times the gameplay seems like a cross
between Grand Theft Auto and Ecco the Dolphin. You
can go on killing sprees, chowing down on anything
that moves including seals, dolphins and people. If
youre not particularly hungry you can throw
them around or ram then with a nose attack. You can
also take out your great white aggression on objects
in the marina such as boats, docks and various personal
watercraft. There is no shortage of things, people
and places to interact with including some astounding
boss fights featuring other demons of the deep including
killer whales.
main problem with the game, and yes there is more
than one problem, is the claustrophobic camera angles
that obscures whatever peripheral vision you are lucky
to be afforded in a videogame. The environments are
huge and you are heartily encouraged to free roam
but on many occasions the camera swings to the most
useless of angles. You end up with tunnel vision.
If you didnt get a good look at the environment
prior to the crazy camera shifts, and you dont
have a photographic memory, youre going to be
out of luck when it comes to avoiding an object, harpoon
or another potentially dangerous situation.
problem is that the shark can get hung up on invisible
parts of objects. Since they are impossible to see,
they are impossible to avoid. The best advice I can
give you is to stay far away from all edges. If you
get hung up, and you most likely will in more than
a few instances, you will find that there are plenty
of times that you are unable to free yourself. The
only way out is to shut-er-down and start the
level all over again. I call this procedure: Reload,
replay, regret, repeat.
of the objectives can be very challenging. Im
not entirely sure if I should blame the controls or
just attribute the lack of precision to the fact that
you are attempting to make a three-thousand pound
shark perform like a trained seal. The missions and
side quests give some structure to the gameplay. There
is a story but its, pardon the pun, pretty shallow.
There are boats that you have to destroy which require
that you pick up speed from a distance and ram them
like a torpedo. Its very difficult to stay your
course with the controls as the shark just isnt
very maneuverable. Not to mention the limiting perspectives
of the camera. With the ability to go below and above
the water surface the camera doesnt always follow
when youre close to the surface. If you get
too close to the surface, the camera will try to anticipate
your next move and give you an above-water perspective
even though youre still underwater. The same
holds true when you are above water. There is no happy
medium with the camera; the only thing you can do
is try to live with it.
or Bruce, also has the ability to swish his tail which
can inflict some damage. But probably the most dramatic
of his moves is his ability to jump directly onto
dry land and snatch an unsuspecting victim. You have
a few seconds to wriggle back into the water before
you expire. This move is so ludicrous that I wouldnt
be surprised if Huckleberry Hound walked out of the
bushes and began doing a narrative on shark hunting.
ridiculous scenarios ensue when you are trapped inside
a theme park. You will actually have to find ways
to secure human hostages so that you can swipe them
across sensors to gain access to other areas. You
will also kill killer whales in an attempt to unlock
other doors. Exploding barrels can be picked up in
your mouth and launched at an intended target. Apparently
the only things that Jaws cant do are climb
a ladder and sneak past guards wearing a disguise.
That's for the sequel: Tom Clancy's Splinter Shark.
is a darn good looking shark and many of the underwater
environments are good looking as well. But this should
be expected since the developers are responsible for
Ecco the Dolphin. There is a good amount of detail
on land but there are also some really bad graphics
that make the humans look like buoys or decoys. Its
so frustrating it makes you just want to take them
in your mouth, shake your head violently and go Argggghhhh
all for something different, but the fact that the
developers dont even attempt to acknowledge
that their tongue is planted firmly in their cheeks
makes me think that they expect us to take this game
seriously. Theres not enough mind-altering chemicals
on the planet to make that a reality. Taking this
game seriously is akin to committing intellectual
suicide. I cant even believe that this game
is tied into the Jaws license. I wont say that
the game drags the license down into the depths, but
it barely treads water with it. Maybe in Jaws 2 we
can just hunt the shark with Huckleberry Hound.
Cole Smith
Senior Writer
"Do you like fish? Well, he likes you too..."
Nearly 30 years after the original motion picture
Jaws was released in theaters, the great white killer
makes its triumphant return, this time to the small
Appaloosa, better known for their Ecco the Dolphin
series, return to the deep blue with "Jaws Unleashed".
For those who remember the 1987 version of "Jaws"
from LJN Ltd, worry not, this isn't the same boat
chase. Instead of searching for Jaws in the immortal
ship, The Orca, players take on the role of Jaws itself.
You will have the power to use Jaws' 3,000 teeth to
consume, well just about anything.
coolest and most alluring aspect of this game is the
possibility of endless amounts of violence. After
all, Jaws has a strict diet, anything it can get its
teeth locked onto. To recreate the mortal destruction
that the shark exhibited in the movies, Appaloosa
created a custom dismemberment engine that feature
up to 64 dismemberment points. Tasty humans aren't
the only things Jaws can get a hold of. Pretty much
every object or obstacle can be destroyed. Just like
the humans, the objects in game (such as boats, docks,
harbors and subs) all have multiple break points,
allowing to you cause damage to any part of an object.
You can wreak as much havoc as you can with underwater
attacks, ocean surface attacks, or you can even use
Jaws' power to launch him up into the air for jumping
Luckily this isn't just a bite 'em up shark game branded
with the Jaws license. The game takes place 30 years
after the first movie and includes some of its themes
and locales. Characters from the film are said to
have been included. There is no set goal when taking
on the role of the deadly fish. The game gives you
about 20 missions, which range from time trials to
destruct-a-thons. It is said that Appaloosa also included
boss battles.
"Jaws Unleashed" looks pretty good. The
background of the free roaming world come into perspective
the closer you get to them. The Jaws model and skins
look very dead on, while the people and vehicles look
like that of the "Grand Theft Auto" series.
A difficult task for game artists to recreate is realistic
water. Games like "FarCry" have achieved
this feat however, and Jaws can be added to the list.
Being out into the deep ocean, the water is reflective
on the surface and crystal clear when underwater.
Light even grows darker the deeper you go.
If you aren't afraid of the water, watch out for this
game's August 16th, 2005 release.
Rating Legend
1.0 - 1.9 = Avoid
2.0 - 2.4 = Poor
2.5 - 2.9 = Average
3.0 - 3.4 = Fair
3.5 - 3.9 = Good
4.0 - 4.4 = Great
4.5 - 4.9 = Must Buy
5.0 = The Best