Validation and method comparison for a point-of-care lateral flow assay measuring equine whole blood insulin concentrations

The Wellness Ready Test (WRT) is a lateral flow, stall-side assay that measures equine insulin in whole blood and requires validation before recommending clinical use. We evaluated intra- and inter-assay precision and linearity and compared the WRT with a radioimmunoassay (RIA). Tested concentrations ranged from <139 to >695 pmol/L (<20 to >100 μIU/mL). For 20 replicates at each insulin level, intra-assay CVs of the WRT for insulin were 13.3%, 12.9%, and 15.3% at low (139–278 pmol/L; 20–40 μIU/mL), intermediate (278–417 pmol/L; 40–60 μIU/mL), and high (>417  pmol/L; >60 μIU/mL) concentrations, respectively. For 10 replicates at each level (3 assay lots), inter-assay CVs were 15.9%, 11.0%, and 11.7%, respectively. In the weighted linear regression of 5 measured insulin concentrations against expected concentrations, R² = 0.98, slope = 1.02, and y-intercept = 14.4 pmol/L (2.08 μIU/mL). The Spearman correlation coefficient (r_s) was 0.90 (95% CI: 0.85–0.94) between the WRT and RIA; the WRT = f(RIA) Passing–Bablok regression yielded the fit, y = 1.005x + 24.3 pmol/L (3.50 μIU/mL). The WRT result averaged 10.4% higher than the RIA result, with targeted bias of 25.9, 26.1, and 26.7 pmol/L (3.74, 3.76, and 3.84 μIU/mL) for cutoffs used to diagnose insulin dysregulation of 312, 347, and 451 pmol/L (45, 50, and 65 μIU/mL). Assay clinical sensitivities, specificities, and accuracies determined at the 3 selected clinical cutoffs and using the RIA as gold standard were 87–95%, 92–96%, and 91–95%, respectively (n = 99 samples). Observed total error was 28.4–30.4%. The WRT had acceptable precision, excellent linearity, and good association with the RIA.     

Winter curing of <Emphasis Type="Italic">Prunus dulcis</Emphasis> cv ‘Butte,’ <Emphasis Type="Italic">P. webbii</Emphasis> and their interspecific hybrid in response to <Emphasis Type="Italic">Xylella fastidiosa</Emphasis> infections

In traditional quantitative genetics, additive effects of genes acting in a population of biparental homozygous lines are estimated on the basis of the phenotypic observations only, usually by taking a difference between mean values for extreme lines. Current molecular methods allow to estimate the additive effects by additionally taking into account the marker data. In this paper we compare these two methods of estimation of additive gene action effects analytically, by simulations and by analysis of real data sets for doubled haploid lines and recombinant inbred lines. The analytic comparison shows under which conditions an agreement of the two methods can be achieved. In most of the considered experimental data and in simulations we observe that the additive effect calculated on the basis of the marker observations is smaller than the total additive effect obtained from phenotypic observations only. This result is discussed, and a weighted regression approach is proposed as a method which can close the gap between the purely phenotypic and genotypic approaches.     

Genetic diversity of Korean Bacillus anthracis isolates from soil evaluated with a single nucleotide repeat analysis

Bacillus (B.) anthracis, the etiological agent of anthrax, is one of the most genetically monomorphic bacteria species in the world. Due to the very limited genetic diversity of this species, classification of isolates of this bacterium requires methods with high discriminatory power. Single nucleotide repeat (SNR) analysis is a type of variable-number tandem repeat assay that evaluates regions with very high mutation rates. To subtype a collection of 21 isolates that were obtained during a B. anthracis outbreak in Korea, we analyzed four SNR marker loci using nucleotide sequencing analysis. These isolates were obtained from soil samples and the Korean Center for Disease Control and Prevention. The SNR analysis was able to detect 13 subgenotypes, which allowed a detailed evaluation of the Korean isolates. Our study demonstrated that the SNR analysis was able to discriminate between strains with the same multiple-locus variable-number tandem repeat analysis genotypes. In summary, we obtained SNR results for four SNR marker loci of newly acquired strains from Korea. Our findings will be helpful for creating marker systems and help identify markers that could be used for future forensic studies.     

Changes in fineroot growth dynamics in response to phosphorus application in an Acacia mangium plantation in Vietnam

Fineroots (≤ 2 mm diameter) are dynamic components of the forest ecosystems and play important role in water and nutrient acquisition in forests. These roots are sensitive to forest fertilization and therefore, the response of fineroots to nutrient application would provide better understanding of the forest carbon and nutrient dynamics that will be helpful in sustainable forest management plans. Two fertilization treatments, including (1) F400: 400 g P₂O₅ (16.5%)/tree and (2) F600: 600 g P₂O₅ /tree, and F0: control (without fertilization), were applied in an Acacia mangium plantation with a planting density of 1100 trees/ha (3 m?×?3 m). The evaluation of fineroot growth across seasons showed that fertilization significantly increased production and subsequent mortality and decomposition. The total decomposition associated with F600 application was 7.95 tons ha^?1 year^?1, equaling 121% of F400 and 160% of the control. Mortality in F600 was 8.75 tons ha^?1 year^?1, equaling 111% of F400 and 198% of F0, while production in F600 was 10.40 tons ha^?1 year^?1, equaling 127% of F400 and 143% of F0. Fineroot production, mortality, and decomposition are seasonally dependent, with higher values measured in the rainy season than in the dry season. Stand basal area increment was significantly correlated with fineroot production (R²?=?0.75), mortality (R²?=?0.44), and decomposition (R²?=?0.48). This study showed that fertilization could facilitate fineroot production, which can then lead to a higher turnover of carbon and nutrients through the decomposition of the greater mass of the fineroots.

     

Magnetic resonance imaging appearance of hypertensive encephalopathy in a dog

A 16-year-old female spayed English Staffordshire terrier was presented for evaluation of a 10-month history of intermittent myoclonic episodes, and a one weeks history of short episodes of altered mentation, ataxia and collapse. Magnetic resonance imaging identified subcortical oedema, predominately in the parietal and temporal lobes and multiple cerebral microbleeds.Serum biochemistry, indirect blood pressure measurements and magnetic resonance imaging abnormalities were consistent with hypertensive encephalopathy secondary to chronic kidney disease.     

Histopathology and oxidative stress analysis of concomitant misoprostol and celecoxib administration

Nonsteroidal anti-inflammatory drugs (NSAIDs), non-selective or selective inhibitors of cyclooxygenase (COX-1 and -2), reduce pain and inflammation associated with arthritic diseases. Celecoxib, a COX-2-selective inhibitor providing decreased gastric injury relative to non-selective NSAIDs, is commonly prescribed. Misoprostol, a prostaglandin analog, supplements NSAID-inhibited prostaglandin levels. As concomitant celecoxib and misoprostol administration has been shown to intensify renal adverse effects, this article examined the influence of concomitant administration on hepatic histopathology, oxidative stress, and celecoxib concentration. On days 1 and 2, rat groups (n = 6) were gavaged twice daily (two groups with vehicle and two groups with 100 μg/kg misoprostol). From day 3 to day 9, one celecoxib dose (40 mg/kg) replaced a vehicle dose of one group and one group received celecoxib in addition to misoprostol. Livers were harvested on day 10. No hepatic abnormalities were observed denoting a lack of influence by either drug. Also no change in mean biomarker levels was detected. The changes in hepatic celecoxib concentration in the misoprostol-receiving group compared to control were not significant. Thus misoprostol does not influence hepatic celecoxib effects in terms of histopathology, oxidative stress, or celecoxib concentration level at the dosage and duration examined.     

What does "build research on farmer practice" mean? Rice crop establishment (beusani) in eastern India as an illustration

In recent years agricultural scientists have been encouraged to build upon farmer practice and knowledge. Although some researchers continued to view farmer practice as irrelevant, others responded with: a) experiments using “farmer practice” as a treatment, b) attempts to transform practices, c) research to better understand practices, and d) research to improve productivity of practices. Researchers generally have not reached the point of testing farmer practices in new areas in anticipation of wider extension. A widespread eastern Indian farmers' practice provides an example: beusani combines direct dry seeded rice, wet plowing at 20–35 days after emergence, laddering, and seedling redistribution as a set of techniques to loosen soils, thin seedlings and improve tillering, control weeds, and optimize plant stand. Results of a survey of the practice are presented; and work on or related to beusaniis reviewed to see how research has built upon farmer practice and how the process can be improved.     

Beach nourishment: an ecologically sound coastal defence alternative? A review

• 1. Even though beach nourishment is generally considered as an environment‐friendly option for coastal protection and beach restoration, sizeable impacts on several beach ecosystem components (microphytobenthos, vascular plants, terrestrial arthropods, marine zoobenthos and avifauna) are described in the literature, as reviewed in this paper.
• 2. Negative, ecosystem‐component specific effects of beach nourishment dominate in the short to medium term, with the size of the impact being determined by (1) activities during the construction phase, (2) the quality and (3) the quantity of the nourishment sand, (4) the timing, place and size of project, and (5) the nourishment technique and strategy applied. Over the long term the speed and degree of ecological recovery largely depend on the physical characteristics of the beach habitat, mainly determined by (1) sediment quality and quantity, (2) the nourishment technique and strategy applied, (3) the place and the size of nourishment and (4) the physical environment prior to nourishment.
• 3. The limited information available on indirect and cumulative ecological effects indicates that these effects cannot be neglected in an overall impact assessment. Hence, for ecologically good practice of beach nourishment it is advised (1) to choose nourishment sands with a sediment composition comparable to that of the natural sediment, (2) to avoid short‐term compaction by ploughing immediately after construction, (3) to execute the nourishment in a period of low beach use by birds and other mobile organisms, (4) to choose a number of smaller projects rather than a single large nourishment project and (5) to select the nourishment technique with respect to the local natural values.
• 4. In order to allow an objective, scientifically sound, ecological adjustment of future nourishments, research should aim at (1) taking into account the full sandy beach ecosystem, (2) avoiding strategic imperfections in experimental design and (3) elucidating the biological processes behind impact and recovery of all ecosystem components.

Copyright © 2006 John Wiley & Sons, Ltd.     

Prevalence and risk factors associated with Cryptosporidium oocysts shedding in pigs in Central Vietnam

We investigated the prevalence and risk factors associated with Cryptosporidium oocysts shedding in pigs in Central Vietnam. A total of 740 single fecal samples collected from diarrheic and non-diarrheic pigs on 89 farms were screened by the modified Ziehl-Neelsen staining method. Prevalence at the animal and the farm levels were 18.1% (134/740) and 71.9% (64/89), respectively. Risk factors for the infection were identified using univariate and multivariate logistic regression analysis. The results revealed that age, sanitary condition and topography were significantly associated with oocyst shedding (P<0.05). Pre-weaned piglets were at the highest risk for infection, followed by post-weaners, sows and finishing pigs. Good sanitary conditions showed positive effects in decreasing oocysts shedding. Topographically, Cryptosporidium was more common in mountainous zone than that in coastal delta zone. There was an association between the occurrence of diarrhea and the level of Cryptosporidium oocyst excretion within infected pigs. This is the first epidemiological investigation of prevalence and risk factors of Cryptosporidium in pigs in Vietnam.