Validation of a Low-Cost Human Papillomavirus Genotyping Assay Based on PGMY PCR and Reverse Blotting Hybridization with Reusable Membranes.

The genotyping of human papillomaviruses (HPV) is essential for the surveillance of HPV vaccines. We describe and validate a low-cost PGMY-based PCR assay (PGMY-CHUV) for the genotyping of 31 HPV by reverse blotting hybridization (RBH). Genotype-specific detection limits were 50 to 500 genome equivalents per reaction. RBH was 100% specific and 98.61% sensitive using DNA sequencing as the gold standard (n = 1,024 samples). PGMY-CHUV was compared to the validated and commercially available linear array (Roche) on 200 samples. Both assays identified the same positive (n = 182) and negative samples (n = 18). Seventy-six percent of the positives were fully concordant after restricting the comparison to the 28 genotypes shared by both assays. At the genotypic level, agreement was 83% (285/344 genotype-sample combinations; κ of 0.987 for single infections and 0.853 for multiple infections). Fifty-seven of the 59 discordant cases were associated with multiple infections and with the weakest genotypes within each sample (P < 0.0001). PGMY-CHUV was significantly more sensitive for HPV56 (P = 0.0026) and could unambiguously identify HPV52 in mixed infections. PGMY-CHUV was reproducible on repeat testing (n = 275 samples; 392 genotype-sample combinations; κ of 0.933) involving different reagents lots and different technicians. Discordant results (n = 47) were significantly associated with the weakest genotypes in samples with multiple infections (P < 0.0001). Successful participation in proficiency testing also supported the robustness of this assay. The PGMY-CHUV reagent costs were estimated at $2.40 per sample using the least expensive yet proficient genotyping algorithm that also included quality control. This assay may be used in low-resource laboratories that have sufficient manpower and PCR expertise.

LDL-cholesterol concentrations: a genome-wide association study.

BACKGROUND: LDL cholesterol has a causal role in the development of cardiovascular disease. Improved understanding of the biological mechanisms that underlie the metabolism and regulation of LDL cholesterol might help to identify novel therapeutic targets. We therefore did a genome-wide association study of LDL-cholesterol concentrations. METHODS: We used genome-wide association data from up to 11,685 participants with measures of circulating LDL-cholesterol concentrations across five studies, including data for 293 461 autosomal single nucleotide polymorphisms (SNPs) with a minor allele frequency of 5% or more that passed our quality control criteria. We also used data from a second genome-wide array in up to 4337 participants from three of these five studies, with data for 290,140 SNPs. We did replication studies in two independent populations consisting of up to 4979 participants. Statistical approaches, including meta-analysis and linkage disequilibrium plots, were used to refine association signals; we analysed pooled data from all seven populations to determine the effect of each SNP on variations in circulating LDL-cholesterol concentrations. FINDINGS: In our initial scan, we found two SNPs (rs599839 [p=1.7x10(-15)] and rs4970834 [p=3.0x10(-11)]) that showed genome-wide statistical association with LDL cholesterol at chromosomal locus 1p13.3. The second genome screen found a third statistically associated SNP at the same locus (rs646776 [p=4.3x10(-9)]). Meta-analysis of data from all studies showed an association of SNPs rs599839 (combined p=1.2x10(-33)) and rs646776 (p=4.8x10(-20)) with LDL-cholesterol concentrations. SNPs rs599839 and rs646776 both explained around 1% of the variation in circulating LDL-cholesterol concentrations and were associated with about 15% of an SD change in LDL cholesterol per allele, assuming an SD of 1 mmol/L. INTERPRETATION: We found evidence for a novel locus for LDL cholesterol on chromosome 1p13.3. These results potentially provide insight into the biological mechanisms that underlie the regulation of LDL cholesterol and might help in the discovery of novel therapeutic targets for cardiovascular disease.

Elaboración de un video sobre calidad, seguridad y medio ambiente en la Unidad de Laboratorios Docentes (ULD) de la Facultad de Farmacia de la Universidad de Barcelona

La Unidad de Laboratorios Docentes (ULD) de la Facultad de Farmacia (UB) ha implantado un sistema de gestión de la calidad (SGC) que permite transmitir al estudiante una formación adicional con el objetivo de mejorar sus competencias transversales (siguiendo las directrices del Espacio Europeo de Educación Superior, EEES). Además, en el curso académico 06-07, se instauraron las Buenas Prácticas Ambientales (BPAL) para disponer de un sistema de gestión que incorporase los criterios de calidad, medioambiente y seguridad (sistema de gestión integrada, SGI). Durante el presente curso académico se ha procedido a la grabación y edición de un video sobre calidad, seguridad y medioambiente en la ULD con el objetivo de mejorar la formación transversal de los estudiantes facilitando la integración de los conocimientos y habilidades profesionales. De esta forma, al salir del entorno universitario, los licenciados y graduados de la Facultad de Farmacia dispondrán de un valor añadido en su formación, mejorando así sus competencias para el desarrollo de su futura profesión. En el video se muestra la manera de trabajar correctamente según las normas de calidad, seguridad y medioambiente recogidas además en un tríptico que se entrega a los estudiantes al acceder por primera vez a un laboratorio de prácticas. El video se difundirá a través de la página web de la ULD, de la videoteca de la UB, del canal You Tube Canal UB, así como de las asignaturas que lo soliciten (sirviendo de soporte para el personal docente).

Los hábitats del proyecto CORINE en el ámbito territorial catalán: delimitación y cartografía.

nd items. Once maps are drawn up they are digitized and put into a GIS and finally subjected to quality control. Table 4 shows the most important habitats (according to polygon number and area covered) and the least represented habitats in the sheets drawn so far. Sheets can be obtained through internet (www.gencat.net/mediamb/pn.htm).

Ablation of human colon carcinoma in nude mice by 131I-labeled monoclonal anti-carcinoembryonic antigen antibody F(ab')2 fragments.

Pooled F(ab')2 fragments of three MAbs against distinct epitopes of carcinoembryonic antigen (CEA) were used for radioimmunotherapy of nude mice bearing a subcutaneous human colon carcinoma xenograft. 9-10 d after transplantation when tumor nodules were in exponential growth, 36 mice were treated by intravenous injection of different amounts of 131I-labeled MAb F(ab')2. All 14 mice injected with a single dose of 2,200 (n = 10) or 2,800 microCi (n = 4) showed complete tumor remission. 8 of the 10 mice treated with 2,200 microCi survived in good health for 1 yr when they were killed and shown to be tumor free. Four of nine other mice treated with four fractionated doses of 400 microCi showed no tumor relapse for more than 9 mo. In contrast, all 15 mice injected with 1,600-3,000 microCi 131I-control IgG F(ab')2 showed tumor growth retardation of only 1-4 wk, and 15 of 16 mice injected with unlabeled anti-CEA MAb F(ab')2 showed unmodified tumor progression as compared with untreated mice. From tissue radioactivity distributions it was calculated that by an injection of 2,200 microCi 131I-MAb F(ab')2 a mean dose of 8,335 rad was selectively delivered to the tumor, while the tissue-absorbed radiation doses for the normal organs were: peripheral blood, 2,093; stomach, 1,668; kidney, 1,289; lung, 1,185; liver, 617; spleen, 501; small intestine, 427; large intestine, 367; bone, 337; and muscle, 198. These treatments were well tolerated since out of 19 mice with complete tumor remission only 4 required bone marrow transplantation and 17 were in good health for 6-12 mo of observation. The results demonstrate the selective destruction of established human colon carcinoma transplants by intravenous injection of either single or fractionated doses of 131I-MAb F(ab')2.

Elaboración de un video sobre calidad, seguridad y medio ambiente en la Unidad de Laboratorios Docentes (ULD) de la Facultad de Farmacia de la Universidad de Barcelona

La Unidad de Laboratorios Docentes (ULD) de la Facultad de Farmacia (UB) ha implantado un sistema de gestión de la calidad (SGC) que permite transmitir al estudiante una formación adicional con el objetivo de mejorar sus competencias transversales (siguiendo las directrices del Espacio Europeo de Educación Superior, EEES). Además, en el curso académico 06-07, se instauraron las Buenas Prácticas Ambientales (BPAL) para disponer de un sistema de gestión que incorporase los criterios de calidad, medioambiente y seguridad (sistema de gestión integrada, SGI). Durante el presente curso académico se ha procedido a la grabación y edición de un video sobre calidad, seguridad y medioambiente en la ULD con el objetivo de mejorar la formación transversal de los estudiantes facilitando la integración de los conocimientos y habilidades profesionales. De esta forma, al salir del entorno universitario, los licenciados y graduados de la Facultad de Farmacia dispondrán de un valor añadido en su formación, mejorando así sus competencias para el desarrollo de su futura profesión. En el video se muestra la manera de trabajar correctamente según las normas de calidad, seguridad y medioambiente recogidas además en un tríptico que se entrega a los estudiantes al acceder por primera vez a un laboratorio de prácticas. El video se difundirá a través de la página web de la ULD, de la videoteca de la UB, del canal You Tube Canal UB, así como de las asignaturas que lo soliciten (sirviendo de soporte para el personal docente).

A LC-tandem MS assay for the simultaneous measurement of new antiretroviral agents: Raltegravir, maraviroc, darunavir, and etravirine.

Raltegravir (RAL), maraviroc (MVC), darunavir (DRV), and etravirine (ETV) are new antiretroviral agents with significant potential for drug interactions. This work describes a sensitive and accurate liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of plasma drug levels. Single-step extraction of RAL, MVC, DRV, ETV and RTV from plasma (100 microl) is performed by protein precipitation using 600 microl of acetonitrile, after the addition of 100 microl darunavir-d(9) (DRV-d(9)) at 1000 ng/ml in MeOH/H(2)O 50/50 as internal standard (I.S.). The mixture is vortexed, sonicated for 10 min, vortex-mixed again and centrifuged. An aliquot of supernatant (150 microl) is diluted 1:1 with a mixture of 20 mM ammonium acetate/MeOH 40/60 and 10 microl is injected onto a 2.1 x 50 mm Waters Atlantis-dC18 3 microm analytical column. Chromatographic separations are performed using a gradient program with 2 mM ammonium acetate containing 0.1% formic acid and acetonitrile with 0.1% formic acid. Analytes quantification is performed by electrospray ionisation-triple quadrupole mass spectrometry using the selected reaction monitoring detection in the positive mode. The method has been validated over the clinically relevant concentrations ranging from 12.5 to 5000 ng/ml, 2.5 to 1000 ng/ml, 25 to 10,000 ng/ml, 10 to 4000 ng/ml, and 5 to 2000 ng/ml for RAL, MRV, DRV, ETV and RTV, respectively. The extraction recovery for all antiretroviral drugs is always above 91%. The method is precise, with mean inter-day CV% within 5.1-9.8%, and accurate (range of inter-day deviation from nominal values -3.3 to +5.1%). In addition our method enables the simultaneous assessment of raltegravir-glucuronide. This is the first analytical method allowing the simultaneous assay of antiretroviral agents targeted to four different steps of HIV replication. The proposed method is suitable for the Therapeutic Drug Monitoring Service of these new regimen combinations administered as salvage therapy to patients having experienced treatment failure, and for whom exposure, tolerance and adherence assessments are critical.

Determination of the Optimum Base Characteristics for Pavements, TR-482, 2004

In recent years, it has become apparent that the design and maintenance of pavement drainage extends the service life of pavements. Most pavement structures now incorporate subsurface layers. Part of the function of these subsurface layers is to drain away excess water, which can be extremely deleterious to the life of the pavement. To assure the effectiveness of such drainage layers after they have been spread and compacted, simple, rapid, in-situ permeability and stability testing and end-result specification are needed. This report includes conclusions and recommendations related to four main study objectives: (1) Determine the optimal range for in-place stability and in-place permeability based on Iowa aggregate sources; (2) Evaluate the feasibility of an air permeameter for determining the permeability of open and well-graded drainage layers in situ; (3) Develop reliable end-result quality control/quality assurance specifications for stability and permeability; and (4) Refine aggregate placement and construction methods to optimize uniformity.

X-Ray systems: radiation protection program in basic research applications

Although the radiation doses involved in basic research radiology are relatively small, the increasing number of radiological procedures makes risks becoming increasingly high. Quality control techniques in radiological practice have to ensure an adequate system of protection for people exposed to radiation. These techniques belong to a quality assurance program for X-ray machines and are designed to correct problems related to equipment and radiological practices, to obtain radiological images of high quality and to reduce the unnecessary exposures.

The Use of Ternary Mixtures in Concrete, TPF-5(117), 2014

This manual is a summary of the findings of a comprehensive study. Its purpose is to provide engineers with the information they need to make educated decisions on the use of ternary mixtures for constructing concrete structures. It discusses the effects of ternary mixtures on fresh and hardened mixture properties and on concrete sustainability; factors that need to be considered for both structural and mixture design; quality control issues; and three example mixtures from constructed projects

Quantitative ultrasound in the management of osteoporosis: the 2007 ISCD Official Positions

Dual-energy X-ray absorptiometry (DXA) is commonly used in the care of patients for diagnostic classification of osteoporosis, low bone mass (osteopenia), or normal bone density; assessment of fracture risk; and monitoring changes in bone density over time. The development of other technologies for the evaluation of skeletal health has been associated with uncertainties regarding their applications in clinical practice. Quantitative ultrasound (QUS), a technology for measuring properties of bone at peripheral skeletal sites, is more portable and less expensive than DXA, without the use of ionizing radiation. The proliferation of QUS devices that are technologically diverse, measuring and reporting variable bone parameters in different ways, examining different skeletal sites, and having differing levels of validating data for association with DXA-measured bone density and fracture risk, has created many challenges in applying QUS for use in clinical practice. The International Society for Clinical Densitometry (ISCD) 2007 Position Development Conference (PDC) addressed clinical applications of QUS for fracture risk assessment, diagnosis of osteoporosis, treatment initiation, monitoring of treatment, and quality assurance/quality control. The ISCD Official Positions on QUS resulting from this PDC, the rationale for their establishment, and recommendations for further study are presented here.

CleanEx: a database of heterogeneous gene expression data based on a consistent gene nomenclature.

The main goal of CleanEx is to provide access to public gene expression data via unique gene names. A second objective is to represent heterogeneous expression data produced by different technologies in a way that facilitates joint analysis and cross-data set comparisons. A consistent and up-to-date gene nomenclature is achieved by associating each single experiment with a permanent target identifier consisting of a physical description of the targeted RNA population or the hybridization reagent used. These targets are then mapped at regular intervals to the growing and evolving catalogues of human genes and genes from model organisms. The completely automatic mapping procedure relies partly on external genome information resources such as UniGene and RefSeq. The central part of CleanEx is a weekly built gene index containing cross-references to all public expression data already incorporated into the system. In addition, the expression target database of CleanEx provides gene mapping and quality control information for various types of experimental resource, such as cDNA clones or Affymetrix probe sets. The web-based query interfaces offer access to individual entries via text string searches or quantitative expression criteria. CleanEx is accessible at: http://www.cleanex.isb-sib.ch/.

Implantació d’un sistema APPCC en les línies de canelons, croquetes i lasanyes a l’empresa de plats precuinats CUINATS JOTRI S.L.

Les diferents normatives, europea i nacional regulen la necessitat d’establir sistemes d’autocontrolque assegurin un nivell minim de seguretat dels productes alimentaris i de les normes relatives alsmanipuladors d’aliments. Les empreses del sector alimentari son les responsables de la higiene en elsseus establiments, basant-se en l’Analisi de Perills i Punts de Control Critic (APPCC) i controlant elpossible risc contra la salut en la cadena alimentaria. Els responsables de l’empresa de platsprecuinats cuinats Jotri S.L. estan interessats en la implementacio d’un sistema d’autocontrol de laqualitat. Per aixo estan interessats en la implementacio d’un sistema d’autocontrol de la seguretatalimentaria (APPCC). Es realitzara la determinacio de punts critics a controlar en les linies de canelonsde carn i l’apliacio de mesures preventives i mesures correctores en les linies de canelons, croquetesi lasanyes

Video Inspection of Highway Edgedrain Systems, HR-317, 1998

Minimizing infiltration of water in pavement structures has long been a priority of pavement designers. Incorporation of subsurface edgedrains is frequently an integral part of an pavement drainage system. In order for such a system to be effective however, it must be properly installed and maintained. With advances in video technology, inspection of edgedrain systems can now be conducted quite efficiently. This report documents the results of 287 video inspections of highway edgedrain systems in 29 states. These inspections were conducted to both demonstrate the capabilities of the technology as well as demonstrating some of the common problems associated with the performance of edgedrain systems. Findings indicated not only that the equipment was quite effective in identifying edgedrain performance concerns, but also how widespread the concerns of edgedrain performance are. Almost one third of the systems inspected had nonfunctional outlets, another third were either found to have non-functional mainlines or the mainlines could not be inspected due to physical obstructions. Only one third of the systems inspected were found to be performing as intended. Recommendations are provided for edgedrain design improvements to facilitate performance of the system and their inspections as well as recommendations to improve quality control during construction. Suggestions are also provided for maintenance procedures to address concerns identified in the inspection process. A Draft Guide Specification For Video Edgedrain Inspection and Acceptance is also provided as an Appendix.

Investigation of Rapid Thermal Analysis Procedures for Prediction of the Service Life of PCCP Carbonate Coarse Aggregate, HR-337, 1993

The major objective of this research project was to use thermal analysis techniques in conjunction with x-ray analysis methods to identify and explain chemical reactions that promote aggregate related deterioration in portland cement concrete. Twenty-two different carbonate aggregate samples were subjected to a chemical testing scheme that included: • bulk chemistry (major, minor and selected trace elements) • bulk mineralogy (minor phases concentrated by acid extraction) • solid-solution in the major carbonate phases • crystallite size determinations for the major carbonate phases • a salt treatment study to evaluate the impact of deicer salts Test results from these different studies were then compared to information that had been obtained using thermogravimetric analysis techniques. Since many of the limestones and dolomites that were used in the study had extensive field service records it was possible to correlate many of the variables with service life. The results of this study have indicated that thermogravimetric analysis can play an important role in categorizing carbonate aggregates. In fact, with modern automated thermal analysis systems it should be possible to utilize such methods on a quality control basis. Strong correlations were found between several of the variables that were monitored in this study. In fact, several of the variables exhibited significant correlations to concrete service life. When the full data set was utilized (n = 18), the significant correlations to service life can be summarized as follows ( a = 5% level): • Correlation coefficient, r, = -0.73 for premature TG loss versus service life. • Correlation coefficient, r, = 0.74 for relative crystallite size versus service life. • Correlation coefficient, r, = 0.53 for ASTM C666 durability factor versus service life. • Correlation coefficient, r, = -0.52 for acid-insoluble residue versus service life. Separation of the carbonate aggregates into their mineralogical categories (i.e., calcites and dolomites) tended to increase the correlation coefficients for some specific variables (r sometimes approached 0.90); however, the reliability of such correlations was questionable because of the small number of samples that were present in this study.