Assessment of recent HIV-1 infection by a line immunoassay for HIV-1/2 confirmation

BACKGROUND: Knowledge of the number of recent HIV infections is important for epidemiologic surveillance. Over the past decade approaches have been developed to estimate this number by testing HIV-seropositive specimens with assays that discriminate the lower concentration and avidity of HIV antibodies in early infection. We have investigated whether this "recency" information can also be gained from an HIV confirmatory assay. METHODS AND FINDINGS: The ability of a line immunoassay (INNO-LIA HIV I/II Score, Innogenetics) to distinguish recent from older HIV-1 infection was evaluated in comparison with the Calypte HIV-1 BED Incidence enzyme immunoassay (BED-EIA). Both tests were conducted prospectively in all HIV infections newly diagnosed in Switzerland from July 2005 to June 2006. Clinical and laboratory information indicative of recent or older infection was obtained from physicians at the time of HIV diagnosis and used as the reference standard. BED-EIA and various recency algorithms utilizing the antibody reaction to INNO-LIA's five HIV-1 antigen bands were evaluated by logistic regression analysis. A total of 765 HIV-1 infections, 748 (97.8%) with complete test results, were newly diagnosed during the study. A negative or indeterminate HIV antibody assay at diagnosis, symptoms of primary HIV infection, or a negative HIV test during the past 12 mo classified 195 infections (26.1%) as recent (< or = 12 mo). Symptoms of CDC stages B or C classified 161 infections as older (21.5%), and 392 patients with no symptoms remained unclassified. BED-EIA ruled 65% of the 195 recent infections as recent and 80% of the 161 older infections as older. Two INNO-LIA algorithms showed 50% and 40% sensitivity combined with 95% and 99% specificity, respectively. Estimation of recent infection in the entire study population, based on actual results of the three tests and adjusted for a test's sensitivity and specificity, yielded 37% for BED-EIA compared to 35% and 33% for the two INNO-LIA algorithms. Window-based estimation with BED-EIA yielded 41% (95% confidence interval 36%-46%). CONCLUSIONS: Recency information can be extracted from INNO-LIA-based confirmatory testing at no additional costs. This method should improve epidemiologic surveillance in countries that routinely use INNO-LIA for HIV confirmation.

Blood donor screening: how to decrease the risk of transfusion-transmitted hepatitis B virus?

QUESTIONS UNDER STUDY: The risk of transfusion-transmitted HBV remains significant in Switzerland, where routine screening for hepatitis B virus (HBV) in blood donations relies solely on serological hepatitis B surface antigen (HBsAg) testing. This study was designed to determine the prevalence of anti-hepatitis B core (anti-HBc) and HBV nucleic acid testing (NAT) positive donations in two different Swiss donor populations, to help in deciding whether supplemental testing may bring additional safety to blood products. METHODS: In a first population of donors, 18143 consecutive donations were screened initially for HBsAg, anti-HBc (with one EIA assay) and with HBV NAT in minipools of 24 donations. The screening repeatedly reactive anti-HBc donations were then "confirmed" with two supplemental anti-HBc assays, an anti-hepatitis B surface assay (anti-HBs) and with single donation HBV NAT. In a second population of donors, 4186 consecutive donations were screened initially with two different anti-HBc assays in addition to the mandatory HBsAg screening test. The screening repeatedly reactive donations with at least one anti-HBc assay were tested for anti-HBs. RESULTS: In the first subset of 18143 donations, 17593 (97.0%) were negative for HBsAg, anti-HBc and HBV NAT in minipools. 549 (3.0%) were HBsAg and HBV NAT negative, but repeatedly reactive for anti-HBc. Of these 549 donations, 287 could not be "confirmed" with two additional anti-HBc assays and were negative with an anti-HBs assay, as well as with single donation HBV NAT. Only 211 (1.2% of the total screened donations) were "confirmed" positive with at least one of two supplemental anti-HBc assays. One repeatedly reactive HBsAg donation, from a first-time donor, was confirmed positive for HBsAg and anti-HBc, as well as with single donation HBV NAT. In the second subset of 4186 donations, 4014 (95.9%) were screened negative for HBsAg and for anti-HBc, tested with two independent anti-HBc assays. 172 donations (4.1%) were HBsAg negative but repeatedly reactive with at least one of the two anti-HBc assays. Of these 172 samples, 86 were reactive with the first anti-HBc assay only, 13 were reactive with the second anti-HBc assay only and 73 (1.7% of the total screened donations) were "confirmed" positive with both anti-HBc assays. CONCLUSION: The prevalence of anti-HBc "confirmed" positive donations in the two Swiss blood donor populations studied was low (<2%) and we found only one HBV NAT positive (HBsAg positive) donation among more than 18000. Concerning blood product safety, an increase in the deferral rate of less than 2% of anti-HBc positive, potentially infectious donors, would in our opinion make routine anti-HBc testing of blood donations cost-effective. There is however still a need for more specific assays to avoid an unacceptably high deferral rate of "false" positive donors. In contrast, the introduction of HBV NAT in minipools gives minimal benefit due to the inadequate sensitivity of the assay. It remains to evaluate more extensively the value of individual donation NAT, alone or in addition to anti-HBc, as supplemental testing in the context of several Swiss blood donor populations.

U.S. utilities' experiences with the implementation of energy efficiency programs

In the U.S., many electric utility companies are offering demand-side management (DSM) programs to their customers as ways to save money and energy. However, it is challenging to compare these programs between utility companies throughout the U.S. because of the variability of state energy policies. For example, some states in the U.S. have deregulated electricity markets and others do not. In addition, utility companies within a state differ depending on ownership and size. This study examines 12 utilities’ experiences with DSM programs and compares the programs’ annual energy savings results that the selected utilities reported to the Energy Information Administration (EIA). The 2009 EIA data suggests that DSM program effectiveness is not significantly affected by electricity market deregulation or utility ownership. However, DSM programs seem to generally be more effective when administered by utilities located in states with energy savings requirements and DSM program mandates.

Determination of ethyl glucuronide in human serum by capillary zone electrophoresis and an immunoassay

Ethyl glucuronide (EtG) is a marker of recent alcohol consumption. For the optimization of the analysis of EtG by CZE with indirect absorbance detection, the use of capillaries with permanent and dynamic wall coatings, the composition of the BGE, and various sample preparation procedures, including dilution with water, ultrafiltration, protein precipitation, and SPE, were investigated. Two validated screening assays for the determination of EtG in human serum, a CZE-based approach and an enzyme immunoassay (EIA), are described. The CZE assay uses a coated capillary, 2,4-dimethylglutaric acid as an internal standard, and a pH 4.65 BGE comprising 9 mM nicotinic acid, epsilon-aminocaproic acid and 10% v/v ACN. Proteins are removed via precipitation with ACN prior to analysis and the LOQ is 0.50 mg/L. The EIA is based upon commercial reagents which are promoted for the determination of urinary EtG. Krebs-Ringer solution containing 5% BSA is used as a calibration matrix. All samples are ultrafiltered prior to analysis of the ultrafiltrate on a Mira Plus analyzer. Assay calibration ranged between 0 and 2 mg/L and the upper reference limit was determined to be 0.05 mg/L. Both assays proved to be suitable for the analysis of samples from different individuals. For EtG levels above 0.50 mg/L, good agreement was observed for the comparison of the results of the two methods.

Simultaneous quantitative detection of relevant biomarkers in breast cancer by quantitative real-time PCR

The assessment of ERa, PgR and HER2 status is routinely performed today to determine the endocrine responsiveness of breast cancer samples. Such determination is usually accomplished by means of immunohistochemistry and in case of HER2 amplification by means of fluorescent in situ hybridization (FISH). The analysis of these markers can be improved by simultaneous measurements using quantitative real-time PCR (Qrt-PCR). In this study we compared Qrt-PCR results for the assessment of mRNA levels of ERa, PgR, and the members of the human epidermal growth factor receptor family, HER1, HER2, HER3 and HER4. The results were obtained in two independent laboratories using two different methods, SYBR Green I and TaqMan probes, and different primers. By linear regression we demonstrated a good concordance for all six markers. The quantitative mRNA expression levels of ERa, PgR and HER2 also strongly correlated with the respective quantitative protein expression levels prospectively detected by EIA in both laboratories. In addition, HER2 mRNA expression levels correlated well with gene amplification detected by FISH in the same biopsies. Our results indicate that both Qrt-PCR methods were robust and sensitive tools for routine diagnostics and consistent with standard methodologies. The developed simultaneous assessment of several biomarkers is fast and labor effective and allows optimization of the clinical decision-making process in breast cancer tissue and/or core biopsies.

Changing ‘Multi-Problem Families’ – Developing a Multi-Contextual Systemic Approach

Over the past 30 years the Marlborough Family Service in London has pioneered multi-family work with marginalized families presenting simultaneously with abuse and neglect, family violence, substance misuse, educational failure and mental illness. The approach is based on a systemic multi-contextual mode and this chapter describes the evolving work, including the establishment of the first permanent multiple family day setting, specifically designed for and solely dedicated to the work with seemingly ‘hopeless’ families. The ingredients of ‘therapeutic assessments’ of parents and families are outlined and the importance of initial network meetings with professionals and family members is emphasized.

Kinetics of maternally acquired anti-hepatitis A antibodies: prediction of waning based on maternal or cord blood antibody levels

BACKGROUND Timing is critical for efficient hepatitis A vaccination in high endemic areas as high levels of maternal IgG antibodies against the hepatitis A virus (HAV) present in the first year of life may impede the vaccine response. OBJECTIVES To describe the kinetics of the decline of anti-HAV maternal antibodies, and to estimate the time of complete loss of maternal antibodies in infants in León, Nicaragua, a region in which almost all mothers are anti-HAV seropositive. METHODS We collected cord blood samples from 99 healthy newborns together with 49 corresponding maternal blood samples, as well as further blood samples at 2 and 7 months of age. Anti-HAV IgG antibody levels were measured by enzyme immunoassay (EIA). We predicted the time when antibodies would fall below 10 mIU/ml, the presumed lowest level of seroprotection. RESULTS Seroprevalence was 100% at birth (GMC 8392 mIU/ml); maternal and cord blood antibody concentrations were similar. The maternal antibody levels of the infants decreased exponentially with age and the half-life of the maternal antibody was estimated to be 40 days. The relationship between the antibody concentration at birth and time until full waning was described as: critical age (months)=3.355+1.969 × log(10)(Ab-level at birth). The survival model estimated that loss of passive immunity will have occurred in 95% of infants by the age of 13.2 months. CONCLUSIONS Complete waning of maternal anti-HAV antibodies may take until early in the second year of life. The here-derived formula relating maternal or cord blood antibody concentrations to the age at which passive immunity is lost may be used to determine the optimal age of childhood HAV vaccination.

International horse movements and spread of equine diseases: Equine Infectious Anaemia and Glanders - two examples

International trade with horses is important and continuously increasing. Therefore the risk of spread of infectious diseases is permanently present. Within this context the worldwide situation of equine vector-borne diseases and of other diseases which are notifiable to the World Organisation of Animal Health (OIE), is described. Furthermore it provides estimates of the numbers of horse movements between these countries, as well as information on import requirements and preventive measures for reducing the risk of disease spread. According to TRACES (Trade Control and Expert System of the European Union) data from 2009 and 2010 81 horses per week were imported from North America into Europe, 42 horses per week from South America, 11 horses per week from the North of Africa and the African horse sichness free-zone of South Africa, 28 per week from the Middle East and the rest of Asia and approximately 4 horses per week from Australia / Oceania. Trade within the European Union resulted amongst others in the introduction of Equine Infectious Anaemia (EIA) from Roma- nia into other European countries. Another example is the suspected case of glanders which occurred after importation of horses from Leb- anon via France and Germany into Switzerland in July 2011.

PREPARATION AND CHARACTERIZATION OF COVALENTLY LINKED ENZYME-ANTIBODY CONJUGATES AND THEIR USE IN MEASURING MINUTE QUANTITIES OF PROTEIN ANTIGENS

The discovery and characterization of oncofetal proteins have led to significant advances in early cancer diagnosis and therapeutic monitoring of patients undergoing cancer chemotherapy. These tumor-associated antigens are presently measured by sensitive, specific immunoassay techniques based on the detection of minute amounts of labeled antigen or antibody incorporated into immune complexes, which must be isolated from free antigen and antibody.^ Since there are several disadvantages with using radioisotopes, the most common immunolabel, one major objective was to prepare covalently coupled enzyme-antibody conjugates and evaluate their use as a practical alternative to radiolabeled immune reagents. An improved technique for the production of enzyme-antibody conjugates was developed that involves oxidizing the carbohydrate moieties on a glycoprotein enzyme, then introducing antibody in the presence of polyethylene glycol (PEG). Covalent enzyme-antibody conjugates involving alkaline phosphatase and amyloglucosidase were produced and characterized.^ In order to increase the sensitivity of detecting the amyloglucosidase-antibody conjugate, an enzyme cycling assay was developed that measures glucose, the product of maltose cleavage by amyloglucosidase, in the picomole range. The increased sensitivity obtained by combined usage of the amyloglucosidase-antibody conjugate and enzyme cycling assay was then compared to that of conventional enzyme immunoassay (EIA).^ For immune complex isolation, polystyrene tubes and protein A-bearing Staphylococcus aureus were evaluated as solid phase matrices, upon which antibodies can be immobilized. A sandwich-type EIA, using antibody-coated S. aureus, was developed that measures human albumin (HSA) in the nanogram range. The assay, using an alkaline phosphatase-anti-HSA conjugate, was applied to the determination of HSA in human urine and evaluated extensively for its clinical applicability.^ Finally, in view of the clinical significance of alpha-fetoprotein (AFP) as an oncofetal antigen and the difficulty with its purification for use as an immunogen and assay standard, a chemical purification protocol was developed that resulted in a high yield of immunochemically pure AFP. ^

Prevalence estimation of tick-borne encephalitis virus (TBEV) antibodies in dogs from Finland using novel dog anti-TBEV IgG MAb-capture and IgG immunofluorescence assay based on recombinant TBEV subviral particles

Tick-borne encephalitis (TBE) is one of the most dangerous human neurological infections occurring in Europe and Northern parts of Asia with thousands of cases and millions vaccinated against it. The risk of TBE might be assessed through analyses of the samples taken from wildlife or from animals which are in close contact with humans. Dogs have been shown to be a good sentinel species for these studies. Serological assays for diagnosis of TBE in dogs are mainly based on purified and inactivated TBEV antigens. Here we describe novel dog anti-TBEV IgG monoclonal antibody (MAb)-capture assay which is based on TBEV prME subviral particles expressed in mammalian cells from Semliki Forest virus (SFV) replicon as well as IgG immunofluorescence assay (IFA) which is based on Vero E6 cells transfected with the same SFV replicon. We further demonstrate their use in a small-scale TBEV seroprevalence study of dogs representing different regions of Finland. Altogether, 148 dog serum samples were tested by novel assays and results were compared to those obtained with a commercial IgG enzyme immunoassay (EIA), hemagglutination inhibition test and IgG IFA with TBEV infected cells. Compared to reference tests, the sensitivities of the developed assays were 90-100% and the specificities of the two assays were 100%. Analysis of the dog serum samples showed a seroprevalence of 40% on Åland Islands and 6% on Southwestern archipelago of Finland. In conclusion, a specific and sensitive EIA and IFA for the detection of IgG antibodies in canine sera were developed. Based on these assays the seroprevalence of IgG antibodies in dogs from different regions of Finland was assessed and was shown to parallel the known human disease burden as the Southwestern archipelago and Åland Islands in particular had considerable dog TBEV antibody prevalence and represent areas with high risk of TBE for humans.

La Evaluación de impacto ambiental y el ordenamiento territorial

En general las metodologías utilizadas para la evaluación de impacto ambiental han empleado ponderaciones subjetivas, o mediciones de ciertos componentes en forma parcializada, en donde no se tiene en cuenta el territorio sobre el cual se desarrollan las acciones y los efectos. Esto hace que las EIA no sean ajustadas, tornándose más dificultoso aún en las evaluaciones relacionadas con temas de Ordenamiento Territorial, donde la complejidad está dada no solo por la cantidad de variables a incluir sino por el nivel de interacciones a considerar. A pesar de que no siempre es tenida en cuenta, la componente espacial es fundamental en toda EIA, no solo para describir el entorno en el que las acciones se realizan sino para el análisis y correlación de fenómenos que impactan en el territorio y el medio ambiente. En este trabajo se persigue: • Identificar indicadores geográficos posibles de incorporar en las EIA. • Medir en forma cuali y cuantitativa los indicadores seleccionados . • Utilizar geotecnologías como son los SIG y la Teledetección en dicha medición. La incorporación de indicadores geográficos en la EIA de temas tan complejos y comprometidos como los relacionados con el Ordenamiento Territorial se torna necesario debido a la propia naturaleza de ciertos fenómenos que requieren de un enfoque integral georeferenciado, con indicadores medidos en forma cualitativa o cuantitativa.

Estudio comparativo entre la Primera Circunscripción Judicial y la Tercera Circunscripción Judicial

La base de la investigación radica en comparar dos perspectivas de intervención en el proceso de adopción que se implementan en la provincia: una de ellas es la que se lleva a cado en el RUA (Registro Único de Adopción) de Capital (Primera Circunscripción Judicial) y por otro lado, la que se implementa por parte de los profesionales del Departamento de Gral. San Martín (Tercera Circunscripción Judicial). La finalidad de tal comparación es indagar cómo se lleva a cabo el proceso de adopción en ambas circunscripciones, desde la instancia en la que quienes desean convertirse en padres adoptivos se inscriben, hasta la integración definitiva del niño/niña en la nueva familia. En este proceso se tendrán en cuenta el cúmulo de representaciones sociales que giran en torno a la temática y las distintas formas de abordar y entender a la adopción en cuanto a la intervención profesional El puntapié de tal indagación, parte de conocer que la Primera Circunscripción Judicial cuenta con un EIA (Equipo Interdisciplinario de Adopción) que aborda el proceso de manera integral y está especializado en la temática; mientras que en la Tercera el trabajo se realiza con los profesionales del Juzgado de manera multidisciplinaria es decir, cada profesional desde su especialidad aborda la temática sin confrontar perspectivas a lo largo de todo el proceso. La importancia de abordar esta temática, radica en comenzar a problematizar las distintas y en algunos casos fragmentarias formas de intervenir en la práctica de la adopción, en las distintas circunscripciones de la provincia. Además, se presume que en torno a la misma giran una serie de representaciones sociales, mitos, prejuicios, no sólo por parte de la sociedad en general, sino también por parte de profesionales que se encargan de abordar esta práctica. Por lo que se apunta a abordar los aspectos básicos que hacen a la adopción, con el fin de garantizar que las intervenciones profesionales se centren en esos criterios.

A avaliação ambiental estratégica como ferramenta ao planejamento e à gestão do território

Planned to be a process of early evaluation of Politics, Plans and Programs (PPPs), which interfere in the environment; the Strategic Environmental Assessment (SEA) suggests a methodology for planning and managing the land, which overcomes the limitations of the traditional plans that try to mitigate the environmental impacts of Projects, assuming a pro-active conception that incorporates the social and environmental aspects in the planning stage of the PPPs. This kind of Evaluation surpasses the existing limitations of the Environmental Impact Assessment (EIA), once the EIA happens after the planning process, when lots of decisions have been made and carried out. In order to overcome the limitations of EIA, the SEA is a strategic tool in the process of planning and managing the land. When we focus on the PPPs and not on the Projects, the SEA, which is more political than technical, priorizes the strategies that assure the integration of the environmental, social, economic and institutional aspects into the planning process, in private or public organizations. In this context, this work aims to establish the concept basis of the Strategic Environmental Evaluation as a tool for land planning and managing. The methodology procedures used here lie in the literature review concerning the SEA, analyzing how this tool can be introduced as an alternative for sustainable development. Although the SEA is a tool that introduces the sustainable development theme as a guiding principle of planning, it is seldom used by managers and decision makers, locally and nationally

A avaliação ambiental estratégica como ferramenta ao planejamento e à gestão do território

Planned to be a process of early evaluation of Politics, Plans and Programs (PPPs), which interfere in the environment; the Strategic Environmental Assessment (SEA) suggests a methodology for planning and managing the land, which overcomes the limitations of the traditional plans that try to mitigate the environmental impacts of Projects, assuming a pro-active conception that incorporates the social and environmental aspects in the planning stage of the PPPs. This kind of Evaluation surpasses the existing limitations of the Environmental Impact Assessment (EIA), once the EIA happens after the planning process, when lots of decisions have been made and carried out. In order to overcome the limitations of EIA, the SEA is a strategic tool in the process of planning and managing the land. When we focus on the PPPs and not on the Projects, the SEA, which is more political than technical, priorizes the strategies that assure the integration of the environmental, social, economic and institutional aspects into the planning process, in private or public organizations. In this context, this work aims to establish the concept basis of the Strategic Environmental Evaluation as a tool for land planning and managing. The methodology procedures used here lie in the literature review concerning the SEA, analyzing how this tool can be introduced as an alternative for sustainable development. Although the SEA is a tool that introduces the sustainable development theme as a guiding principle of planning, it is seldom used by managers and decision makers, locally and nationally

A avaliação ambiental estratégica como ferramenta ao planejamento e à gestão do território

Planned to be a process of early evaluation of Politics, Plans and Programs (PPPs), which interfere in the environment; the Strategic Environmental Assessment (SEA) suggests a methodology for planning and managing the land, which overcomes the limitations of the traditional plans that try to mitigate the environmental impacts of Projects, assuming a pro-active conception that incorporates the social and environmental aspects in the planning stage of the PPPs. This kind of Evaluation surpasses the existing limitations of the Environmental Impact Assessment (EIA), once the EIA happens after the planning process, when lots of decisions have been made and carried out. In order to overcome the limitations of EIA, the SEA is a strategic tool in the process of planning and managing the land. When we focus on the PPPs and not on the Projects, the SEA, which is more political than technical, priorizes the strategies that assure the integration of the environmental, social, economic and institutional aspects into the planning process, in private or public organizations. In this context, this work aims to establish the concept basis of the Strategic Environmental Evaluation as a tool for land planning and managing. The methodology procedures used here lie in the literature review concerning the SEA, analyzing how this tool can be introduced as an alternative for sustainable development. Although the SEA is a tool that introduces the sustainable development theme as a guiding principle of planning, it is seldom used by managers and decision makers, locally and nationally