Search

A significant problem confronting Operators in modern field development and production is that created by loss of functionality of the surface controlled subsurface safety valve (SCSSV) due to blockage of, or damage to, the hydraulic control line. The consequent loss of hydraulic pressure to the valve means its closure, the resultant loss of production and the need to provide an alternative safety system in order to continue exploiting the wells reserves Currently two alternatives are available: A full scale workover to pull tubing, replace the inoperable control line and restore functionality to the SCSSV.Installation of a velocity or dome charged subsurface controlled subsurface safety valve (SSCSSV). The former approach requires a major expense, which may not be justifiable in a mature well, while the reliability offered by the latter approach typically does not meet well integrity requirements and can sometimes lead to reduced production. Nederlandse Aardolie Maatschappij (NAM) has been faced with this problem in many onshore and offshore wells and was determined to find an alternative solution. Based on the successful completion of a previous development project, NAM asked Weatherford to jointly develop a solution to this problem, which would allow the installation of an alternative control line without the expense of a workover. In this paper the authors will review the problems associated with loss of control line functionality and consequent SCSSV malfunction. They will go on to describe in detail the joint project which led to the development, testing, and eventual field deployment of the Weatherford Damaged Control Line (WDCL) Safety Valve featuring surface control, which can be installed using wireline and capillary string techniques.

The eukaryotic replisome is a crucial determinant of genome stability, but its structure is still poorly understood. We found previously that many regulatory proteins assemble around the MCM2-7 helicase at yeast replication forks to form the replisome progression complex (RPC), which might link MCM2-7 to other replisome components. Here, we show that the RPC associates with DNA polymerase alpha that primes each Okazaki fragment during lagging strand synthesis. Our data indicate that a complex of the GINS and Ctf4 components of the RPC is crucial to couple MCM2-7 to DNA polymerase alpha. Others have found recently that the Mrc1 subunit of RPCs binds DNA polymerase epsilon, which synthesises the leading strand at DNA replication forks. We show that cells lacking both Ctf4 and Mrc1 experience chronic activation of the DNA damage checkpoint during chromosome replication and do not complete the cell cycle. These findings indicate that coupling MCM2-7 to replicative polymerases is an important feature of the regulation of chromosome replication in eukaryotes, and highlight a key role for Ctf4 in this process.

The neighborhood supermarket shopping center is an increasingly important element of the suburban retail landscape. An analysis of patronage differentials for such centers in San Antonio, TX, reveals that both competitive share and retail mix factors are important. Correlation analysis of a patronage density index with locational and management variables indicates that center and store visibility, distance from a regional mall, retail mix (offering of goods appropriate to a small center), and size of center (smaller is better) were the most important explanatory factors in center viability.

The purpose of this study is to analyze the differences in the economic bases of two small towns, each tributary to nearby metropolitan areas, on either side of the U.S.-Mexico border. Thus, it combines the policy concerns of small town economic development (Alguire et al. 1987) with a social scientific interest in cross-cultural differences. Small towns face certain common problems independent of which side of the border they are on - social stratification, limited shopping opportunities, non-dynamic leaders (more ambitious residents have frequently invested their talents elsewhere), limited employment opportunities and anemic tax bases. Nevertheless, the international border does impose cultural and structural differences which are reflected in differences in the flows of capital, labor and consumption expenses. For example, Mexican rural society is poorer, less residentially mobile, less diurnally mobile and more socially tied to the local community than is the case in the U.S. Also, jobs in government service and income from transfer payments are less available in the average Mexican town than in the average U.S. town. Because of these differences people in the Mexican town are likely to purchase lower-order goods and services, spend more money locally and be less directly dependent on the government or on private pensions, for their livelihood, than their counterparts in the U.S. town. Understanding the fundamental patterns of income flows into and out of the community is a task that economic base analysis is uniquely designed to meet. Here, a modified form of the Tiebout (1962) procedure will be used to trace the basic income flow into and out of establishments and households. A town's economic viability will depend on the magnitude of the difference between the inward and outward flows. The study will analyze a town in south Texas and a town of similar characteristics in northern Coahuila. Economic base analysis in this study will refer both to the study of such exogenous income, and to the study of household and establishment expenditure patterns which determine the degree of income recycling within the local region (the multiplier).

Infectious bronchitis virus (IBV) is the causative agent of avian infectious bronchitis, which is characterized by respiratory, reproductive, and renal signs. However, the role of IBV as an enteric pathogen in still controversial. In Brazil, antigenic groups of IBV divergent from the Massachusetts serotype used for vaccination schedules in that country have already been demonstrated. The present study aimed to assess the different genotypes of IBV in Brazilian commercial poultry flocks by partial sequencing of the S1 amino-terminus coding region using enteric contents as samples and examine their relationship with the vaccine serotype currently in use. Samples of enteric contents were taken as pools of five birds from each of 18 poultry farms (17 broiler and one laying farm) from five Brazilian states between 2002 and 2006. Birds were presenting watery diarrhea and poor general condition but were without respiratory, renal, or reproductive signs. Conventional antibacterial and anticoccidial therapies were unsuccessful and, furthermore, all samples proved negative for rotavirus, reovirus, and astrovirus. Eleven IBV samples were isolated in embryonated eggs and resulted in S1 sequences. Phylogenetic analysis showed that these segregated into an exclusive cluster, close to serotype D274, but distant from Massachusetts. Mean amino acid identity amongst these Brazilian strains was 94.07%; amongst these and serotypes D274, 4/91, and Massachusetts, mean amino acid identity was 77.17%, 69.94%, and 68.93%, respectively. In conclusion, the presence of genotype variant strains of IBV in Brazilian poultry flocks has been demonstrated and might be the reason for the unsuccessful control of IBV in Brazil. Furthermore, these results also strengthen the implications of IBV in enteric diseases of poultry.

Mycobacterium vanbaalenii PYR-1 is well known for its ability to degrade a wide range of high-molecular-weight (HMW) polycyclic aromatic hydrocarbons (PAHs). The genome of this bacterium has recently been sequenced, allowing us to gain insights into the molecular basis for the degradation of PAHs. The 6.5 Mb genome of PYR-1 contains 194 chromosomally encoded genes likely associated with degradation of aromatic compounds. The most distinctive feature of the genome is the presence of a 150 kb major catabolic region at positions 494 approximately 643 kb (region A), with an additional 31 kb region at positions 4,711 approximately 4,741 kb (region B), which is predicted to encode most enzymes for the degradation of PAHs. Region A has an atypical mosaic structure made of several gene clusters in which the genes for PAH degradation are complexly arranged and scattered around the clusters. Significant differences in the gene structure and organization as compared to other well-known aromatic hydrocarbon degraders including Pseudomonas and Burkholderia were revealed. Many identified genes were enriched with multiple paralogs showing a remarkable range of diversity, which could contribute to the wide variety of PAHs degraded by M. vanbaalenii PYR-1. The PYR-1 genome also revealed the presence of 28 genes involved in the TCA cycle. Based on the results, we proposed a pathway in which HMW PAHs are degraded into the beta-ketoadipate pathway through protocatechuate and then mineralized to CO2 via TCA cycle. We also identified 67 and 23 genes involved in PAH degradation and TCA cycle pathways, respectively, to be expressed as proteins.

By rapidly depleting each of the essential budding yeast proteins of unknown function, we identified a novel factor that we call Inn1, which associates with the contractile actomyosin ring at the end of mitosis and is needed for cytokinesis. We show that Inn1 has a C2 domain at the amino terminus of the protein that is required for ingression of the plasma membrane, whereas the remainder of the protein recruits Inn1 to the actomyosin ring. The lethal effects of deleting the INN1 gene can be suppressed by artificial fusion of the C2 domain to other components of the actomyosin ring, restoring membrane ingression on contraction of the actomyosin ring. Our data indicate that recruitment of the C2 domain of Inn1 to the contractile actomyosin ring is crucial for ingression of the plasma membrane during cytokinesis in budding yeast.

Live attenuated subtype B avian metapneumovirus (aMPV) vaccines from two different commercial sources (Vac 1 and Vac 2) were used to vaccinate two groups of day-old specific-pathogen-free chicks. The chicks were challenged at 21 or 49 days of age with a virulent subtype B virus. Parameters compared were persistence of vaccine viruses, their ability to induce humoral antibody responses, and the protection they offered against virulent challenge. Vac 1 virus was detectable for at least 14 days after vaccination, and Vac 2 virus was detectable for at least 7 days after vaccination. The serologic response after Vac 1 by using a subtype B-based enzyme-linked immunosorbent assay was significantly higher than that induced by Vac 2, even though the titer of the recommended dose had a higher titer than Vac 1. After challenge at 21 or 49 days of age, both vaccines gave 100% protection against clinical disease. For the recovery of challenge virus, one of 10 chicks given the Vac 2 was positive by virus isolation or reverse transcriptase-polymerase chain reaction (RT-PCR) on each occasion except on day 10 after challenge at 49 days. With groups vaccinated with Vac 1, no challenge virus was detected except on day 5 after challenge at 49 days, when one of 10 chickens was positive only by RT-PCR. This study has shown that commercial aMPV vaccines from different sources can provide identical protection in terms of clinical disease after virulent challenge and in their ability to induce humoral antibodies, even though the titer of their recommended doses is widely different.

The pathogenesis of infection involving both infectious bronchitis virus (IBV) and avian metapneumovirus (aMPV) causes reproductive damage in hens after viral replication in the epithelium of the oviduct, resulting in loss of cilia and degeneration and necrosis of the epithelial and glandular cells. Although IBV has been indicated as a possible cause of the formation of calcium stones in the epididymus of roosters, a definitive association has not been confirmed. This report describes the detection of IBV and aMPV in the testes of roosters from a Brazilian poultry broiler breeder's flock with epididymal stones and low fertility. Samples of testis, trachea, and lungs from breeder males aged 57 wk were positive for IBV by reverse transcriptase-polymerase chain reaction (RT-PCR), and virus isolation and testis samples were also positive for aMPV by RT-PCR. The inoculation of testis samples into embryonated chicken eggs via the allantoic cavity resulted in curled, hemorrhagic, and stunted embryos typical of IBV infection. The allantoic fluid was positive by RT-PCR aimed to amplify the region coding for the S1 subunit of the IBV S gene, but it was not positive for aMPV. Sequence analysis of the amplified fragment revealed a close relationship with European IBV genotype D274, previously unreported in Brazil. These results indicate that IBV and perhaps aMPV are likely to have played a role in the pathogenesis of the testicular disease described and should be regarded as factors that can influence male fertility disease in chickens.

Mycobacterium vanbaalenii PYR-1 was the first bacterium isolated by virtue of its ability to metabolize the high-molecular-weight polycyclic aromatic hydrocarbon (PAH) pyrene. We used metabolic, genomic, and proteomic approaches in this investigation to construct a complete and integrated pyrene degradation pathway for M. vanbaalenii PYR-1. Genome sequence analyses identified genes involved in the pyrene degradation pathway that we have proposed for this bacterium. To identify proteins involved in the degradation, we conducted a proteome analysis of cells exposed to pyrene using one-dimensional gel electrophoresis in combination with liquid chromatography-tandem mass spectrometry. Database searching performed with the M. vanbaalenii PYR-1 genome resulted in identification of 1,028 proteins with a protein false discovery rate of o -phthalate and the β-ketoadipate pathway. Proteomic analysis also revealed that 18 enzymes in the pathway were upregulated more than twofold, as indicated by peptide counting when the organism was grown with pyrene; three copies of the terminal subunits of ring-hydroxylating oxygenase (NidAB2, MvanDraft_0817/0818, and PhtAaAb), dihydrodiol dehydrogenase (MvanDraft_0815), and ring cleavage dioxygenase (MvanDraft_3242) were detected only in pyrene-grown cells. The results presented here provide a comprehensive picture of pyrene metabolism in M. vanbaalenii PYR-1 and a useful framework for understanding cellular processes involved in PAH degradation.

An integrated omics approach was undertaken in order to elucidate a systems biology level understanding of the acute hepatotoxcity of valproic acid (VPA). Metabonomics, proteomics and gene expression microarray platforms were employed in this systems biology study. CD-1 female pregnant mice were injected subcutaneously with 600 mg/kg VPA or vehicle control. Urine, serum, and liver tissue were collected at 6, 12, and 24 h after dosing. Principal component analysis (PCA) of the metabonomics data showed clustering of the dosed groups away from the controls for the urine samples. Looser clustering was seen in the other sample sets investigated. However, VPA administration resulted in altered glucose concentrations in urine samples at 12 and 24 h and in aqueous liver tissue extracts at 12 h after VPA administration. Proteomics studies identified two proteins, glycogen phosphorylase and amylo-1,6-glucosidase, which were increased in dosed animals relative to control. Both of these proteins are involved in converting glycogen to glucose. Examination of the expression of 20,000 liver genes did not reveal significantly altered expression at 6, 12, or 24 h after VPA exposure. The combined studies indicated a perturbation in the glycogenolysis pathway following administration of VPA.

Mycobacterium vanbaalenii PYR-1 is able to metabolize a wide range of low- and high-molecular-weight (HMW) polycyclic aromatic hydrocarbons (PAHs). A 20-kDa protein was upregulated in PAH-metabolizing M. vanbaalenii PYR-1 cells compared to control cultures. The differentially expressed protein was identified as a β subunit of the terminal dioxygenase using mass spectrometry. PCR with degenerate primers designed based on de novo sequenced peptides and a series of plaque hybridizations were done to screen the M. vanbaalenii PYR-1 genomic library. The genes, designated nidA3B3 , encoding the α and β subunits of terminal dioxygenase, were subsequently cloned and sequenced. The deduced enzyme revealed close similarities to the corresponding PAH ring-hydroxylating dioxygenases from Mycobacterium and Rhodococcus spp. but had the highest similarity, 61.9%, to the α subunit from Nocardioides sp. strain KP7. The α subunit also showed 52% sequence homology with the previously reported NidA from M. vanbaalenii PYR-1. The genes nidA3B3 were subcloned into the expression vector pET-17b, and the enzyme activity in Escherichia coli cells was reconstituted through coexpression with the ferredoxin (PhdC) and ferredoxin reductase (PhdD) genes of the phenanthrene dioxygenase from Nocardioides sp. strain KP7. The recombinant PAH dioxygenase appeared to favor the HMW PAH substrates fluoranthene, pyrene, and phenanthrene. Several other PAHs, including naphthalene, anthracene, and benz[ a ]anthracene, were also converted to their corresponding cis -dihydrodiols. The recombinant E. coli , however, did not show any dioxygenation activity for phthalate and biphenyl. The upregulation of nidA3B3 in M. vanbaalenii PYR-1 induced by PAHs was confirmed by reverse transcription-PCR analysis.

The components of the replisome that preserve genomic stability by controlling the progression of eukaryotic DNA replication forks are poorly understood. Here, we show that the GINS (go ichi ni san) complex allows the MCM (minichromosome maintenance) helicase to interact with key regulatory proteins in large replisome progression complexes (RPCs) that are assembled during initiation and disassembled at the end of S phase. RPC components include the essential initiation and elongation factor, Cdc45, the checkpoint mediator Mrc1, the Tof1-Csm3 complex that allows replication forks to pause at protein-DNA barriers, the histone chaperone FACT (facilitates chromatin transcription) and Ctf4, which helps to establish sister chromatid cohesion. RPCs also interact with Mcm10 and topoisomerase I. During initiation, GINS is essential for a specific subset of RPC proteins to interact with MCM. GINS is also important for the normal progression of DNA replication forks, and we show that it is required after initiation to maintain the association between MCM and Cdc45 within RPCs.

A live A type avian metapneumovirus (AMPV) vaccine which had been shown to be highly protective and short lived in experimental conditions was found to persist for longer periods in the field and to be associated with disease. Previously other factors such as possible secondary pathogens and management considerations had made it impossible to conclude whether the observed disease was a result of an increase in the vaccine virulence. In this study, an AMPV was isolated from poults on a farm which had been vaccinated with the same live A type vaccine. Full sequencing of the isolate, the vaccine and the vaccine progenitor confirmed its vaccine origin and further showed that generation of the vaccine had only involved nine substitutions of which three coded for amino acid changes. The isolated virus was inoculated into 1-day-old turkey poults in disease secure isolators and shown to cause disease with a severity similar to that caused by virulent field virus. Only two coding mutations were associated with this reversion to virulence.

Multiprotein complexes have been increasingly recognized as essential functional units for a variety of cellular processes, including the protein degradation system. Selective degradation of proteins in eukaryotes is primarily conducted by the ubiquitin proteasome system. The current knowledge base, pertaining to the proteasome complexes in mammalian cells, relies largely upon information gained in the yeast system, where the 26S proteasome is hypothesized to contain a 20S multiprotein core complex and one or two 19S regulatory complexes. To date, the molecular structure of the proteasome system, the proteomic composition of the entire 26S multiprotein complexes, and the specific designated function of individual components within this essential protein degradation system in the heart remain virtually unknown. A functional proteomic approach, employing multidimensional chromatography purification combined with liquid chromatography tandem mass spectrometry and protein chemistry, was utilized to explore the murine cardiac 26S proteasome system. This article presents an overview on the subject of protein degradation in mammalian cells. In addition, this review shares the limited information that has been garnered thus far pertaining to the molecular composition, function, and regulation of this important organelle in the cardiac cells.

Protein profiles of Mycobacterium vanbaalenii PYR-1 grown in the presence of high-molecular-weight polycyclic aromatic hydrocarbons (HMW PAHs) were examined by two-dimensional gel electrophoresis (2-DE). Cultures of M. vanbaalenii PYR-1 were incubated with pyrene, pyrene-4,5-quinone (PQ), phenanthrene, anthracene, and fluoranthene. Soluble cellular protein fractions were analyzed and compared, using immobilized pH gradient (IPG) strips. More than 1000 gel-separated proteins were detected using a 2-DE analysis program within the window of isoelectric point (pI) 4-7 and a molecular mass range of 10-100 kDa. We observed variations in the protein composition showing the upregulation of multiple proteins for the five PAH treatments compared with the uninduced control sample. By N-terminal sequencing or mass spectrometry, we further analyzed the proteins separated by 2-DE. Due to the lack of genome sequence information for this species, protein identification provided an analytical challenge. Several PAH-induced proteins were identified including a catalase-peroxidase, a putative monooxygenase, a dioxygenase small subunit, a small subunit of naphthalene-inducible dioxygenase, and aldehyde dehydrogenase. We also identified proteins related to carbohydrate metabolism (enolase, 6-phosphogluconate dehydrogenase, indole-3-glycerol phosphate synthase, and fumarase), DNA translation (probable elongation factor Tsf), heat shock proteins, and energy production (ATP synthase). Many proteins from M. vanbaalenii PYR-1 showed similarity with protein sequences from M. tuberculosis and M. leprae. Some proteins were detected uniquely upon exposure to a specific PAH whereas others were common to more than one PAH, which indicates that induction triggers not only specific responses but a common response in this strain.

This study describes attempts to increase and measure sensitivity of molecular tests to detect avian pneumovirus (APV). Polymerase chain reaction (PCR) diagnostic tests were designed for the detection of nucleic acid from an A-type APV genome. The objective was selection of PCR oligonucleotide combinations, which would provide the greatest test sensitivity and thereby enable optimal detection when used for later testing of field materials. Relative and absolute test sensitivities could be determined because of laboratory access to known quantities of purified full-length DNA copies of APV genome derived from the same A-type virus. Four new nested PCR tests were designed in the fusion (F) protein (2 tests), small hydrophobic (SH) protein (1 test), and nucleocapsid (N) protein (1 test) genes and compared with an established test in the attachment (G) protein gene. Known amounts of full-length APV genome were serially diluted 10-fold, and these dilutions were used as templates for the different tests. Sensitivities were found to differ between the tests, the most sensitive being the established G test, which proved able to detect 6,000 copies of the G gene. The G test contained predominantly pyrimidine residues at its 3 9 termini, and because of this, oligonucleotides for the most sensitive F test were modified to incorporate the same residue types at their 39 termini. This was found to increase sensitivity, so that after full 3 9 pyrimidine substitutions, the F test became able to detect 600 copies of the F gene. Avian pneumovirus (APV) causes an upper respiratory tract infection in turkeys, leading to turkey rhinotracheitis, and in some other avian species, including chickens where it is also involved in the etiology of multifactorial diseases such as swollen head syndrome. The APVs were first re- ported from South Africa in the late 1970s. 3 Several years

The serine/threonine kinase protein kinase C epsilon (PKC epsilon) has been shown to be a critical component in the heart's resistance to cell death following ischemic insult. Recent studies have indicated that PKC epsilon forms multi-protein signaling complexes to accomplish signal transduction in cardiac protection. Using two-dimensional electrophoresis (2DE), combined with matrix-assisted laser desorption ionization mass spectrometry (MS), the initial analysis of these complexes identified signaling molecules, structural proteins, and stress-activated proteins. The initial analysis, although fruitful, was limited by the number of proteins revealed on the 2D gels. It was also apparent that many known cardiac protective functions of PKC epsilon could not be fully accounted for by the proteins identified in the initial analysis. Here we reported the identification of an additional 57 proteins in PKC epsilon complexes using complimentary separation techniques, combined with high sensitivity MS. These techniques include 2DE or large format 1D SDS-PAGE followed by LC/MS/MS and solution trypsin digestion followed by LC/MS/MS, all of which yielded novel data regarding PKC epsilon protein complexes. Nanoscale LC/MS/MS for the analysis of gel-isolated proteins was performed with sub-femtomole sensitivity. In contrast to 2DE analyses, the identification of proteins from 1D gels was independent of their visualization via staining and allowed for the identification of proteins with high isoelectric points. We found that PKC epsilon complexes contain numerous structural and signaling molecules that had escaped detection by our previous analyses. Most importantly, we identified two new groups of proteins that were previously unrecognized as components of the PKC epsilon complex: metabolism-related proteins and transcription/translation-related proteins.

The rat major histocompatibility complex class Ia allelomorph RT1-A1(c) is a potent ligand for the recently identified inhibitory rLy-49 receptor, STOK-2. With the ultimate objective of studying the interactions of these molecules using structural and functional methods, we undertook a detailed study of its peptide specificity. The study revealed that designing an "ideal peptide" by choosing the most abundant residues in the "binding motif" obtained by pool sequencing does not necessarily yield an optimal binding peptide. For RT1-A1(c), as many as four positions, P2, P4, P5, and P9, were detected as putative anchors. Since this molecule displays a preference for highly hydrophobic peptides, we tested binding of peptides derived from the known leader peptide sequences of other rat histocompatibility complex class I molecules. One such peptide, found to bind well, requiring 1.6 microm peptide to achieve 50% stabilization, was searched for in vivo. Natural RT1-A1(c) binding peptides were purified from rat splenocytes and characterized by mass spectrometry using a combined matrix-assisted laser desorption ionization/time-of-flight and quadrupole time-of-flight approach. Results showed that the signal sequence-derived peptide was not detectable in the purified peptide pool, which was composed of a complex spectrum of peptides. Seven of these self-peptides were successfully sequenced.

SIR, — Since 2002 we have been undertaking a survey at the University of Liverpool to detect the presence and identity of infectious bronchitis virus (ibv) in poultry flocks in various western European countries using reverse transcriptase-pcr technology. This relies on examining viral rna from

Colorectal cancer (CRC) is the second leading cause of cancer related deaths in the United States. A critical component of CRC care is post-surgical monitoring for cancer recurrence. Carcinogenicembryonic antigen (CEA) is a tumor marker for the clinical management of CRC which has the specific utility of monitoring post-operative disease recurrence. After surgery to remove cancerous tissue, the level of CEA in blood can be periodically monitored using an immunoassay. If the levels begin to rise above 6.0 ng/mL there is a high correlation with recurrence of the cancer. CEA, like many tumor markers is a glycoprotein and there is a significant body of work showing protein glycosylation is greatly affected by diseases such as cancer. We are investigating the CEA protein microheterogeneity in effort to improve its performance as a CRC biomarker. This poster presentation outlines our methods for glycoprotein characterization and includes qualitative and quantitative CEA glycoform data. CEA immuno-purified from cancerous liver was purchased from AbCam and processed without further purification. The purified protein was processed by SDS-PAGE on a 4-12% Bis-Tris NuPage gel (Invitrogen) in the MOPS buffer system. A second instance of the purified protein was deglycosylated with PNGaseF (New England Biolabs) and processed using the same SDS-PAGE conditions. The windows corresponding to the modified and unmodified proteins were excised and processed by in-gel digestion using a robot (ProGest, DigiLab) and Promega sequencing grade trypsin. The digested sample was analyzed by nano LC/MS/MS with a Waters NanoAcquity HPLC system interfaced to a ThermoFisher Orbitrap Velos Pro. Peptides were loaded on a trapping column and eluted over a 75µm analytical column at 350nL/min; both columns were packed with Jupiter Proteo resin (Phenomenex). The mass spectrometer was operated in data-dependent mode, with MS performed in the Orbitrap at 60,000 FWHM resolution and MS/MS performed in the dual linear ion traps. The fifteen most abundant ions were selected for MS/MS. Data were processed using bioinformatics tools and manually to identify and validate N-glycosylation sites and associated N-linked glycans. Glycoform quantitation was performed using High Resolution Accurate Mass (HRAM) and when necessary glycosites were confirmed using MS3 analysis. This presentation summarizes a workflow for the identification and quantification of protein glycoforms and shows the combination of immuno-purification, a targeted multiple enzyme approach and high resolution accurate mass data enables the accurate identification of N-glycosites and their glycoforms. The data from these experiments highlight the complexity of CEA glycoforms, for example a single N-linked glycosite on the peptide LQLSNGNR has over 30 associated glycoforms of varying abundance. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1267. doi:1538-7445.AM2012-1267

Abstract A general method for the scale up of highly detailed, heterogeneous reservoir cross sections is presented and applied to the simulation of several recovery processes in a variety of geologic settings. The scale up technique proceeds by first identifying portions of the fine scale reservoir description which could potentially lead to high fluid velocities, typically regions of connected, high permeability. These regions are then modeled in detail while the remainder of the domain is coarsened using a general numerical technique for the calculation of effective permeability. The overall scale up method is applied to the cross sectional simulation of three actual fields. Waterflood, steamflood and miscible flood recovery processes are considered. In all these cases, the scale up technique is shown to give coarsened reservoir descriptions which provide simulation results in very good agreement with those of the detailed reservoir descriptions. For these simulations, speedups in computation times, for the coarsened models relative to their fine grid counterparts, range from a factor of 10 to a factor of 200. Introduction Studies documenting the effects of fine scale heterogenous reservoir displacement processes have appeared with increasing frequency over the past several years. One approach to represent such effects in reservoir simulation models is to generate highly detailed geostatistical realizations of rock properties and use these as input to reservoir simulators. However, due to the high level of detail typically included in these geostatistical descriptions, the computational requirements for reservoir simulation can become prohibitive. For example, a detailed three dimensional geostatistical model may contain 10 to 10 grid blocks, while a typical high performance workstation can realistically accommodate reservoir simulation models of only 10 to 10 grid blocks, depending on the type of simulation. Thus, to develop simulation models containing 10 to 10 grid blocks that are representative of highly detailed geologic models containing 10 to 10 grid blocks, a scale up of about a factor of 100 is required. This corresponds to a coarsening of about a factor of 4 or 5 in each linear dimension. We have recently developed and applied a general methodology for the scale up of heterogeneous reservoir cross sections that achieves this degree of coarsening in two coordinate directions. Given a detailed description of a reservoir cross section, the method is designed to generate a coarsened model that will provide predictions for the global pressure - flow rate behavior of the reservoir, the breakthrough characteristics of the displacing fluid and the overall fractional flow curves of the various reservoir fluids, in close agreement with simulation results from the original detailed description. The method proceeds by first identifying regions of potentially high fluid velocity, which are modeled in detail. Then, the bulk of the domain is coarsened using a general technique for the calculation of effective permeability. P. 365^

A prototype live attenuated turkey rhinotracheitis (TRT) vaccine, which was known to cause occasional disease in young poults following multiple back passage, was tested for the presence of a virulent subpopulation by a novel combined in vitro and in vivo screening technique. When vaccine was inoculated at high titres into chick embryo tracheal organ cultures, 17% of aliquots were found to cause ciliostasis, and when these aliquots, in turn, were inoculated into 1-day-old poults, approximately one-quarter caused clinical disease. Removal of the subpopulation by plaque purification led to viruses which had reduced tendency to revert to virulence but remained protective. The technique proved valuable in identifying virulent subpopulations in specific prototype TRT vaccines. The principle may have more general application.

A general method for the scale up of highly detailed, heterogeneous reservoir cross sections to coarser scales, for the purposes of flow simulation of displacement processes, is developed and applied. The technique involves the nonuniform coarsening of the fine scale description, with fine resolution introduced in regions of potentially high fluid velocities (typically regions of connected, high permeability) and coarse, homogenized descriptions applied to the remainder of the reservoir. The method is designed to capture both average behavior as well as some important behaviors which are due to the extremes of the permeability field, such as the breakthrough time of the displacing fluid. The method is applied to several example problems, including two actual field examples, and is shown to provide coarsened models (~ 25 x 25) which give simulation results for fractional flows and saturations in close agreement with fine scale (~ 100 x 100) results. These examples demonstrate the ability of the method to capture a wide variety of flow behavior without the need for specific knowledge of the global flow field, indicating that the coarsened reservoir description is to a large degree process independent.

Although undocumented Mexican immigrants in the United States are highly concentrated in California and Texas their locations have become increasingly dispersed in recent years. The most rapid growth in apprehensions by U.S. authorities is now occurring outside the Southwest. Furthermore the already-dispersed patterns of migrant origins in Mexico are becoming more so over time. Thus the phenomenon of undocumented flows is spreading spatially in both the U.S. and Mexico. Historic factors promoting migrant channelization are being supplanted by factors promoting dispersion: increased migrant job awareness and capability a liberalization of U.S. immigration enforcement and generally worsening conditions in rural Mexico. (EXCERPT)

Most previous attempts to quantify the organizational complexity of visual stimuli have focussed upon stimulus attributes rather than on the perceptual consequences of stimulus processing. We believe that it is more profitable to emphasize perceptual effects and have developed a methodology whereby the smoothness of induced rotation is used to quantify the organizational structure of geometric figures. Our subjects were presented with two-dimensional perspective views of figures in which the azimuth was successively changed. When the tachistoscopic presentation times ranged from 60 to 100 msec, smooth rotation was always perceived for azimuth orders of 0°, 15°, 30°, 30°, 15°, 0°. When a 15° azimuth view was interposed between the two 30° views, an irregularity in rotation was apparent if the presentation duration exceeded a critical value. We used this value as an index of structural detail. Our figures were computer-generated plots of stacked cubes that were produced in either shaded or outline patterns. Subjects viewed two cycles of rotation of either shaded (textured) or outline figures then judged whether the induced rotation was smooth or interrupted. We found that outline figures generally required longer interposition durations than the more solid patterns for rotation to appear interrupted. It seemed as though structural detail, such as texture, provided more information for induced rotation thereby accentuating any flaws in the smoothness of movement.

Honeycomb sheet is already widely used as a core in aeronautical sandwich construction. An alternative application is to use it as the reinforcing element for composites in which the cells of the honeycomb are filled with various materials. This paper presents the results of a study of such a composite, in which a low modulus infill is used. The work covers, in simple terms, the elastic properties of the unfilled sheet and the composite under in-plane direct loading and out-of-plane bending. The plastic deformation characteristics under in-plane direct loading are also considered. Specimen experimental results are presented, which show that the simple analytical approach used is clearly justifiable.

A modification of the Osgood-Tannenbaum sociopsychological model of belief change is applied to University of Cincinnati student perceptions of Southwest Ohio and of the Kentucky Bluegrass. Students scale their beliefs about the two regions; after a week, they are given a factual presentation associating the two regions and asked to again scale their beliefs. The variable which proves most important in the resultant belief changes for the Kentucky Bluegrass is the separation of the two regions on a student's initial belief scale, suggesting the willingness of the students to believe the new information. Further analysis indicates that although objective differences in the regions affect initial beliefs, they play a negligible role in the degree to which students change their beliefs. A “halo effect” is also discovered, in which beliefs not mentioned in the presentation change in predictable directions. The empirical results appear encouraging to recent efforts which have been made to change popular impressions about places.

U.S. job and spatial mobility are compared here for recent returnee migrants from two Mexican areas — Rio Grande, Zacatecas, in the interior; and Nueva Rosita-Muzquiz, Coahuila, near the U.S. border. Results suggest that the interior migrants fit a hierarchical migrant model: they move up the urban hierarchy from U.S. rural areas to towns and cities, experiencing substantial job mobility at first, but little after reaching the urban sector. Border migrants fit a shuttle migrant model: they return to the same job and place year after year, experiencing little or no spatial and occupational mobility, although they tend to hold somewhat higher status jobs.