Effects of hereditary retinal degeneration due to a CEP290 mutation on the feline pupillary light reflex

Objective To understand how progressive rod cone degeneration due to a mutation in CEP290 affects the pupillary light reflex (PLR) in domestic cats. Animals studied Domestic cats identified as either normal wildtype (WT; n = 6), or homozygous for the rdAc mutation in CEP290 and having early stage retinal degeneration (stage 2, S2; n = 4), or advanced retinal degeneration (S4; n = 6). Methods The effect of light on pupil size was measured over a series of 10‐s pulses of white and chromatic light in cats lightly sedated with medetomidine. Results In WT cats, the PLR was characterized by a pronounced initial constriction that rapidly re‐dilated during the stimulus (pupil escape), to a stable or sustained constriction. There was then a marked constriction at stimulus offset. Each component of the PLR was retained in affected cats, but with progressively reduced irradiance sensitivity from early to advanced retinal disease. Conclusions The PLR of cats had multiple phases, with a remarkably high‐amplitude ‘paradoxical’ off‐constriction even in the absence of retinal disease. In rdAc cats, reduced irradiance sensitivity was consistent with progressive loss of rod and cone function. Based on previously characterized retinal pathology, this suggests the visual streak of the retina has a proportionally large contribution to PLR input. These findings support the hypothesis that the efficacy of planned therapeutic trials can be determined by careful evaluation of the PLR in cats.     

Oncogenic CARD11 mutations in human diffuse large B cell lymphoma

Diffuse large B cell lymphoma (DLBCL) is the most common form of non-Hodgkin's lymphoma. In the least curable (ABC) subtype of DLBCL, survival of the malignant cells is dependent on constitutive activation of the nuclear factor-kappaB (NF-kappaB) signaling pathway. In normal B cells, antigen receptor-induced NF-kappaB activation requires CARD11, a cytoplasmic scaffolding protein. To determine whether CARD11 contributes to tumorigenesis, we sequenced the CARD11 gene in human DLBCL tumors. We detected missense mutations in 7 of 73 ABC DLBCL biopsies (9.6%), all within exons encoding the coiled-coil domain. Experimental introduction of CARD11 coiled-coil domain mutants into lymphoma cell lines resulted in constitutive NF-kappaB activation and enhanced NF-kappaB activity upon antigen receptor stimulation. These results demonstrate that CARD11 is a bona fide oncogenein DLBCL, providing a genetic rationale for the development of pharmacological inhibitors of the CARD11 pathway for DLBCL therapy.     

Utilizing technology to enhance laboratory–client interaction while encouraging best behavior

As client interactions with veterinary diagnostic laboratories have evolved, so have client expectations: faster results, enhanced accessibility to cases, and more seamless data transfer from the laboratory database; all of these factors have encouraged the evolution of diagnostic laboratory systems. This evolution started with 24-h access to laboratory results via the web, yet data quality remained at the mercy of the person filling out the form. If bad (incomplete) information was flowing in, then the data coming out was equally bad (incomplete or inconsistent). By designing a web-based system integrated into our existing reporting platform, the Iowa State University Veterinary Diagnostic Laboratory (ISU-VDL) set out to improve the quality of submission data by including the premises identification number (PIN) and obtaining consistent location data, all while presenting to the client an easy-to-use interface. Efforts continued by incentivizing the use of this tool and client submission practices. As clients transitioned, data have become more complete, resulting in easier queries and an improved ability to leverage the diagnostic data. To further enhance the client experience, a streamlined daily reporting summary was designed to communicate laboratory results succinctly. The use of these web-based tools had a positive impact on the quality and consistency of the diagnostic data. As new ideas develop, the ISU-VDL strives to foster continuous improvement and positively impact the clients’ experience.     

Characterization of flowering time and SSR marker analysis of spring and winter type Brassica napus L. germplasm

Flowering dates and life forms of all available Brassica napus accessions conserved at the North Central Regional Plant Introduction Station (NCRPIS) were characterized, and a survey of molecular variation was conducted by using simple sequence repeats (SSR) in order to support better management of accessions with diverse life forms. To characterize flowering phenology, 598 B. napus accessions from the NCRPIS collection were planted in Iowa and Kansas field sites together with a current commercial cultivar and observed for days to flowering (first, 50% and 100% flowering) in 2003. Days from planting to 50% flowering ranged from 34 to 83 in Iowa and from 53 to 89 in Kansas. The mean accumulated growing degree days (GDD) to 50% flowering were 1,997 in Iowa, and 2,106 in Kansas. Between locations, the correlation in flowering time (r = 0.42) and the correlation in computed GDD (r = 0.40) were both significant. Differences in flowering-time rank were observed for several accessions. Accessions that failed to flower in Iowa in a single growing season comprised 28.5% of the accessions; of the flowering accessions, 100% plant flowering was not always achieved. Accessions were grouped according to flowering time. A stratified sample of 50 accessions was selected from these groups, including 10 non-flowering and 40 flowering accessions of diverse geographic origins and phenological variation. The flowering time observed in the sampled accessions when grown in the greenhouse were found to be significantly correlated to the flowering time observed in the field locations in Iowa (r = 0.79) and Kansas (r = 0.49). Thirty SSR markers, selected across 18 Brassica linkage groups from BrassicaDB, and 3 derived from Brassica expressed sequence tags (ESTs) were scored in the stratified sample. An average of three bands per SSR primer pair was observed. Associations of SSR marker fragments with the life forms were determined. Analysis of molecular variation by using cluster analysis and ordination resulted in recognizable, distinct groups of annual and biennial life-form types, which may have direct applications for planning and management of future seed regenerations. Mention of commercial brand names in this paper does not constitute an endorsement of any product by the U.S. Department of Agriculture or cooperating agencies.     

Area-wide management of Aedes albopictus: choice of study sites based on geospatial characteristics, socioeconomic factors and mosquito populations

BACKGROUND: Aedes (Stegomyia) albopictus (Skuse), the Asian tiger mosquito, is an introduced invasive species in the United States that is responsible for a significant proportion of service requests to local mosquito control programs. This container‐utilizing mosquito is refractory to standard mosquito abatement measures in the United States. This study is part of a USDA‐ARS project to develop an area‐wide management strategy for Ae. albopictus. The goal was to identify three study sites, similar in socioeconomic parameters, geography and Ae. albopictus abundance, in urban and suburban areas in Mercer and Monmouth counties in New Jersey. Prior service requests and light trap counts and also detailed county maps were used to chose nine preliminary sites (four in Mercer and five in Monmouth) where weekly surveillance for Ae. albopictus was performed throughout the 2008 active season. RESULTS: Although outliers were detected, socioeconomic variables in the study sites within each county were fairly consistent. Ae. albopictus abundance was associated with poverty levels and had the highest maxima in Mercer, although average mosquito abundance was similar in urban Mercer and suburban Monmouth. CONCLUSION: Three study sites in each county were identified for future studies. The summer‐long surveillance also revealed socioeconomic variables critical for the development of integrated mosquito management. Copyright © 2011 Society of Chemical Industry     

Simultaneous quantification of eight biogenic amine compounds in tuna by matrix solid-phase dispersion followed by HPLC-orbitrap mass spectrometry

A method for the extraction of agmatine, cadaverine, histamine, phenyethylamine, putrescine, tryptamine, tyramine, and urocanic acid from canned tuna and frozen tuna loin matrices by matrix solid-phase dispersion, followed by separation and quantification of these compounds by ultrahigh-performance hydrophilic interaction chromatography (UHPLC-HILIC) with orbitrap mass spectrometric detection, is described. Tuna samples are dispersed in a CN-silica sorbent and eluted with a mixture of aqueous ammonium formate buffer and acetonitrile. Separation and detection are carried out on an Agilent 1200 high-performance liquid chromatograph coupled to a Thermo Exactive orbitrap mass spectrometer, and metformin is used as the internal standard. Spike recoveries are determined across a range of 20-100 ppm for each compound, and the method is validated with respect to linearity, reproducibility, accuracy, and limits of quantitation and detection. The method is demonstrated to be suitable for use in quantifying these target compounds in the studied matrices.