Cloning,expression, purification,and biological activity of five feline type I interferons

Type I interferons (IFN) are important mediators of the host defense against viral infections in mammals. In humans multiple subtypes of IFN-alpha exist, most of which possess antiviral activity. Little is known about the type I IFN genes in cats and the role they may play in feline immunological responses to viruses. We have isolated cDNAs encoding five feline IFN-alpha (feIFN) subtypes that share from 95 to 99% amino acid sequence identity. FeIFN-alpha5 has five additional amino acids inserted at position 139, which are not present in the other four subtypes. Sequence identity of the feIFN proteins encoded by the five clones compared to human IFN-alpha2 is approximately 60%. Unlike most of the human subtypes, each of the five feline IFN sequences has an N-glycosylation recognition site. Expression of all five feIFN-alpha subtypes in Chinese hamster ovary (CHO) cells was confirmed by Western blot analysis, and all resulting proteins were glycosylated. The antiviral activity of each feIFN-alpha subtype produced in transiently transfected CHO cell cultures was tested in vitro. In addition, subtype feIFN-alpha6 was expressed in the yeast, Pichia pastoris. The resulting secreted mature recombinant protein was purified and demonstrated significant antiviral activity and induction of 2',5'-oligoadenylate synthetase activity in vitro.     

MAGNETIC RESONANCE IMAGING CHARACTERISTICS OF THE FOOT IN HORSES WITH PALMAR FOOT PAIN AND CONTROL HORSES

Palmar foot pain is a common cause of lameness. Magnetic resonance imaging (MRI) has the potential to detect damage in all tissues of the equine foot, but an understanding of the differences in magnetic resonance (MR) images between feet from horses with and without palmar foot pain is required. This study aimed to describe MR findings in feet from horses with no history of foot-related lameness, and to compare these with MR findings in horses with lameness improved by palmar digital local analgesia. Thirty-four limbs from horses euthanized with a clinical diagnosis of navicular syndrome (ameness >2 months duration, positive response to palmar digital nerve blocks and absence of other forelimb problems) (Group L), and 25 feet from age-matched horses with no history of foot pain (Group N) were examined. For each anatomic structure, MR signal intensity and homogeneity, size, definition of margins, and relationships with other structures were described. Alterations in MR signal intensity and homogeneity were graded as mild, moderate, or severe and compared between Groups L and N. Results revealed that there were significant differences in MR images between Groups N and L. Multiple moderate-severe MR signal changes were present in 91% of limbs from Group L and moderate (none were graded severe) in 27% of limbs from Group N. In most Group L limbs, more than three structures and frequently six to eight structures were abnormal. Concomitant abnormalities involved most frequently the deep digital flexor tendon, distal sesamoidean impar ligament, navicular bone, collateral sesamoidean ligament, and navicular bursa (with significant associations in severity grade between these structures), sometimes with involvement of the distal interphalangeal joint and/or its collateral ligaments. It was concluded that findings on MR images were different between horses with and without foot pain, and that pain localized to the foot was associated with MR changes in a variety of structures, indicating that damage to several structures may occur concurrently and that MR imaging was useful for evaluation of foot pain.     

Application and field validation of a PCR assay for the detection of Mycoplasma hyopneumoniae from swine lung tissue samples.

A PCR assay was validated for the detection of Mycoplasma hyopneumoniae in porcine lung tissue. The detection limit of the assay was 0.18 colony-forming units/g of lung sample spiked with M. hyopneumoniae. In field validation, 426 pigs from 220 cases were examined for M. hyopneumoniae infection by M. hyopneumoniae PCR and a fluorescent antibody (FA) test. In total, 103 pig lungs (24.2%) were positive in the PCR test, and 69 pig lungs (16.2%) were positive in the FA test, among which, 62 pigs were positive for both PCR and FA test. Most of the PCR-positive but FA test-negative cases had lesions compatible with M. hyopneumoniae infection. With Bayesian modeling, the diagnostic sensitivity and specificity of the PCR were determined to be 97.3% and 93.0%, respectively.     

Efficiency of early selection for rotation-aged wood quality traits in radiata pine

A total of 360 bark-to-bark-through-pith wood strips were sampled at breast height from 180 trees in 30 open-pollinated families from two rotation-aged genetic trials to study inheritance, age-age genetic correlation, and early selection efficiency for wood quality traits in radiata pine. Wood strips were evaluated by SilviScan® and annual pattern and genetic parameters for growth, wood density, microfibril angle (MFA), and stiffness (modulus of elasticity: MOE) for early to rotation ages were estimated. Annual ring growth was the largest between ages 2–5 years from pith, and decreased linearly to ages 9–10. Annual growth was similar and consistent at later ages. Wood density was the lowest near the pith, increased steadily to age 11–15 years, then was relatively stable after these ages. MFA was highest (35°) near the pith and reduced to about 10° at age 10–15 years. MFA was almost unchanged at later ages. MOE increased from about 2.5 GPa near the pith to about 20 GPa at ages 11–15 years. MOE was relatively unchanged at later ages. Wood density and MOE were inversely related to MFA. Heritability increased from zero near the pith and stabilised at ages 4 or 5 for all four growth and wood quality traits (DBH, density, MFA and MOE). Across age classes, heritability was the highest for area-weighted density and MFA, lowest for DBH, and intermediate for MOE. Age-age genetic correlations were high for the four traits studied. The genetic correlation reached 0.8 after age 7 for most traits. Early selection for density, MFA and MOE were very effective. Selection at age 7–8 has similar effectiveness as selection conducted at rotation age for MFA and MOE and at least 80% effective for wood density.     

Strategies to promote healthier food purchases: a pilot supermarket intervention study

OBJECTIVE: To pilot the design and methodology for a large randomised controlled trial (RCT) of two interventions to promote healthier food purchasing: culturally appropriate nutrition education and price discounts. DESIGN: A 12-week, single-blind, pilot RCT. Effects on food purchases were measured using individualised electronic shopping data ('Shop 'N Go' system). Partial data were also collected on food expenditure at other (non-supermarket) retail outlets. SETTING: A supermarket in Wellington, New Zealand. PARTICIPANTS: Eligible customers were those who were the main household shoppers, shopped mainly at the participating store, and were registered to use the Shop 'N Go system. Ninety-seven supermarket customers (72% women; age 40 +/- 9.6 years, mean +/- standard deviation) were randomised to one of four intervention groups: price discounts, nutrition education, a combination of price discounts and nutrition education, or control (no intervention). RESULTS: There was a 98% follow-up rate of participants, with 85% of all reported supermarket purchases being captured via the electronic data collection system. The pilot did, however, demonstrate difficulty recruiting Maori, Pacific and low-income shoppers using the electronic register and mail-out. CONCLUSIONS: This pilot study showed that electronic sales data capture is a viable way to measure effects of study interventions on food purchases in supermarkets, and points to the feasibility of conducting a large-scale RCT to evaluate the effectiveness of price discounts and nutrition education. Recruitment strategies will, however, need to be modified for the main trial in order to ensure inclusion of all ethnic and socio-economic groups.     

When fish shoals meet: outcomes for evolution and fisheries

The mechanisms underlying the social structure of free‐ranging fish shoals have received little attention in comparison to functional studies on shoaling. Recently, however, a number of investigations, both in the marine and in the freshwater environment, have begun to address the underlying mechanisms by concentrating on interactions between free‐ranging shoals. The rates of shoal encounters can influence the opportunities for individual assortment by phenotype and selection of shoal size; act as a constraint on the observed patterns of shoal structure by restricting individual choice behaviour and were found to be high in several small freshwater species (where intershoal distances were small), but lower in marine species where shoals were more dispersed. The duration of encounters may play a role in that it affects both the time available for assessment (of the encountered shoal) as well as that for exchange of individuals. Scarce published information on the outcome of shoal encounters suggests that the outcome of shoal encounters is influenced by shoal composition but not by shoal size. Individual behaviour may have evolved to maintain shoal size when shoals encounter, but when fish populations are depleted by fishing, this trait can exacerbate range and stock collapse. Furthermore, an understanding of the dynamics of shoal encounters has important consequences for the evolution of reciprocal altruism and the transmission of information through social learning within populations. Finally, information on encounter rates between shoals and the number of individuals that are exchanged on such occasions could be important for making predictions about the spread of disease through fish populations.     

High-throughput assay for detection of soybean lipoxygenase-1

A high-throughput assay was developed to detect soybean lipoxygenase 1 (LOX-1) using a multilabel plate reader. The assay was also adapted to a single cell fluorometer. Fluorescein is degraded by linoleic hydroperoxide produced from soybean lipoxygenase and linoleic acid. The decrease in fluorescence is measured over time, and the area-under-the-curve (AUC) is used to quantify the LOX-1 content of soybean extract. A dose-dependent response is seen with varied dilutions of pure LOX enzyme or soybean extracts. Percent recovery was between 97% and 108%, and relative standard deviation was 4.3%. Advantages of the assay include the reduced preparation time of samples and reduced use of reagents in the high-throughput assay. Multiple samples can be measured in a single run with a multilabel plate reader.     

QTL analysis of morphological and developmental traits in the Ler × Cvi population of Arabidopsis thaliana

Drought frequently constrains production of wheat (Triticum aestivum L.), but development of tolerant cultivars is hampered by low heritability for drought tolerance and a lack of effective selection strategies. Our objective was to identify an optimum selection regime for wheat in drought-prone environments. Six-hundred entries derived from 10 crosses were developed by selection under continuous high moisture, alternating high with low moisture, alternating low with high moisture, and continuous low moisture conditions for five generations. The selections were evaluated in two low-yield, a medium-yield, and a high-yield environment in the Yaqui Valley, Sonora, Mexico. The mean performance of entries derived from a particular selection regime was dependant on the stress level of the evaluation environment. Lines developed and selected under continuous high moisture and continuous low-moisture regimes produced the highest mean yields in the low moisture evaluation environment. There was no relationship between continuous selection under either high yielding conditions or low yielding conditions and the mean performance of the resultant lines in their respective high and low yielding evaluation environments. The mean yield of lines selected using the alternating high/low moisture regime as well as the five highest yielding lines were superior in the HY environment, and had similar performance with other regimes under the low yielding evaluation environment. Our results indicate that alternating selection between high and low yielding environments is the most effective way to develop wheat germplasm adapted to environments where intermittent drought occurs. This revised version was published online in July 2006 with corrections to the Cover Date.