Developing an Automated Enzyme Immunoassay for High-Throughput Screening of Bovine Serum Antibodies to Neospora Caninum

An enzyme-linked immunosorbent assay (ELISA) for Neospora caninum antibodies was automated with a robotic workstation, the Beckman Coulter Biomek 2000, to screen 200 bovine sera. Comparing these results with manually run ELISA data, a 95.92% agreement (K = 0.9592) between the two assays was obtained. The automated assay was specific and sensitive with excellent positive and negative predictive values. The results were repeatable and reproducible. The automation flexibility was high and the operation complexity was minimal. High-throughput screening (HTS) for bovine antibodies to Neospora caninum was achieved. The assay was developed according to the internationally recognized ISO17025 standard requirements.     

Chinook salmon and rainbow trout catch and temperature distributions in Lake Ontario

We determined the distributions of Chinook salmon and rainbow trout by describing seasonal mean vertical and bathymetric catch depths from 1997 to 2005 using angler creel surveys. We developed and applied a cross-validated model of Lake Ontario temperatures to determine the water temperatures associated with these distributions. During April, Chinook salmon and rainbow trout were found nearshore at a bathymetric depth of 20 m. However, rainbow trout were caught at shallower vertical depths (4 to 6 m) than Chinook salmon (8 to 10 m). Both species moved deeper and farther offshore during May, June, and July. Vertical catch depths were similar, but rainbow trout were found further offshore (40 to 65 m bathymetric depth) than Chinook salmon (35 to 50 m bathymetric depth) during June, July and August. During September, Chinook salmon moved closer to shore (25 to 35 m bathymetric depth) and to shallower depths (9 to 12 m), consistent with river mouth staging associated with spawning. Rainbow trout remained offshore (45 to 60 m bathymetric depth) in deeper water (11 to 16 m). The species occupied significantly different spatial habitats during April, August, and September. Mean catch temperatures of both species were similar and increased seasonally to 13 to 14 °C during August and September. Rainbow trout were caught at cooler temperatures than Chinook salmon during June and July. The estimated temperature distributions agree with independent field studies but are different then previously assumed in bioenergetic models.     

The Impacts of Atlantic Salmon Stocking on Rainbow Trout in Barnum House Creek,Lake Ontario

We compared the impacts of stocking age-0 Atlantic salmon (Salmo salar) at high and low densities, and no stocking on abundance and growth of age-0 rainbow trout (Oncorhyncus mykiss) in Barnum House Creek, Ontario during 1993 to 2005. A similar stream, Shelter Valley Creek, was chosen as an appropriate reference stream where age-0 Atlantic salmon were not stocked. The catches of age-0 rainbow trout in Barnum House and the reference stream were highly correlated (r = 0.96) during years when no stocking occurred; however, this relationship did not persist in years when Atlantic salmon were stocked. The catch of age-0 rainbow trout in Barnum House Creek was significantly lower under both high (P = 0.00026) and low (P = 0.011) density Atlantic salmon stocking treatments compared with the no stocking treatment. The catches of age-0 rainbow trout and age-0 Atlantic salmon were negatively correlated in Barnum House Creek (r = −0.63). The length of age-0 rainbow trout in Barnum House Creek was depressed significantly (P = 0.004), under the high intensity Atlantic salmon stocking treatment, but not under the low intensity treatment (P = 0.20). In contrast, the length of age-0 rainbow trout in Shelter Valley Creek was unchanged over the same period. Restoration stocking of Atlantic salmon in Lake Ontario tributaries may impact rainbow trout abundance and growth.     

Plastic ball grid array (PBGA) overview

As integrated circuit functionality and clock speed continue to rise, innovative packaging approaches are in great demand. Recently, the plastic ball grid array (PBGA) technology has been gaining industry-wide interest and commitment as the potentially lowest-cost package for high-I/O devices and even for some lower-pincount applications. Drivers include the density advantages of an area array, quickly achieving six-sigma assembly yields with existing assembly equipment, the potential for excellent electrical and thermal performance, along with the traditional low cost of plastic packages. Because some perceived weaknesses are being eradicated, worldwide evaluation of the PBGA has accelerated. Although various aspects of this technology are discussed frequently, an overall assessment is still under development. In this paper, a systematic and comprehensive evaluation of PBGA technology will be described to identify (1) its technical advantages and limitations, (2) unique application areas where PBGA is the package of choice, and (3) current major hurdles for acceptance of PBGA and possible approaches to overcome these problems. The PBGA will be compared with PQFP, CQFP and CBGA in terms of package characteristics and their impact on system assembly. The characteristics include package attributes (i.e., package size, I/O counts and lead pitch), performance (i.e., electrical, thermal) and reliability (moisture). At the system level, solder joint fatigue, board routing, solder assembly yield, solder reparability and board delay are key metrics. The cost implication of various package families will be discussed. By analogy with SMT, the infrastructure for PBGA will take time to develop. The key elements and the current status of this infrastructure will be discussed.     

Psychoanalysis as a natural science.

Despite Freud's unwavering intention that psychoanalysis should conform to the requirements of a natural science, this aim has not been realized. Some analysts hold the aim to have been mistaken. Others believe it can and should be achieved, proposing that traditional metapsychology be replaced by a new conceptual framework utilizing modern concepts not available to Freud. Utilizing data obtained from current findings in developmental psychology based on direct observation of parent-child interaction as well as from therapeutic sessions, a possible framework is sketched using principles derived from ethology, control theory, and human information processing. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Mobility problems of the disabled

In the UK there are approximately one million disabled people included in Social Services Disablement Registers. There are also about 11 million retired people, many of whom suffer from the illnesses of old age: frailty, arthritis and rheumatism. Together, these two groups consitute a substantial proportion of the population which experiences varying degrees of difficulty in moving about the city and thus performing routine tasks (such as shopping or visiting a medical practitioner); frequent tasks such as making a journey to work are proportionately more difficult.     

Automation and ISO 17025 test validation of a Toxoplasma Gondii antibodies enzyme-linked immunosorbent assay

The authors automated an enzyme-linked immunosorbent assay to detect porcine serum antibodies to Toxoplasma gondii. Two thousand swine sera can be assayed in two eight-hour shifts using a robotic workstation. The automated-ELISA programming is not complex and the test configuration is flexible. This high-throughput screening (HTS) a-ELISA can achieve a 10-fold increase (100→1000 tests) in test capacity over the manual method. The assay has been validated according to the requirements of the ISO/IEC 17025 standard. These include repeatability, reproducibility, and optimal threshold value studies. Other requirements are proficiency panel testing, analyst training, standard operation procedure, and equipment certifications.     

Modulation of the Kv1.3 potassium channel by receptor tyrosine kinases

The voltage-dependent potassium channel, Kv1.3, is modulated by the epidermal growth factor receptor (EGFr) and the insulin receptor tyrosine kinases. When the EGFr and Kv1.3 are coexpressed in HEK 293 cells, acute treatment of the cells with EGF during a patch recording can suppress the Kv1.3 current within tens of minutes. This effect appears to be due to tyrosine phosphorylation of the channel, as it is blocked by treatment with the tyrosine kinase inhibitor erbstatin, or by mutation of the tyrosine at channel amino acid position 479 to phenylalanine. Previous work has shown that there is a large increase in the tyrosine phosphorylation of Kv1.3 when it is coexpressed with the EGFr. Pretreatment of EGFr and Kv1.3 cotransfected cells with EGF before patch recording also results in a decrease in peak Kv1.3 current. Furthermore, pretreatment of cotransfected cells with an antibody to the EGFr ligand binding domain (alpha-EGFr), which blocks receptor dimerization and tyrosine kinase activation, blocks the EGFr-mediated suppression of Kv1.3 current. Insulin treatment during patch recording also causes an inhibition of Kv1.3 current after tens of minutes, while pretreatment for 18 h produces almost total suppression of current. In addition to depressing peak Kv1.3 current, EGF treatment produces a speeding of C-type inactivation, while pretreatment with the alpha-EGFr slows C-type inactivation. In contrast, insulin does not influence C-type inactivation kinetics. Mutational analysis indicates that the EGF-induced modulation of the inactivation rate occurs by a mechanism different from that of the EGF-induced decrease in peak current. Thus, receptor tyrosine kinases differentially modulate the current magnitude and kinetics of a voltage-dependent potassium channel.