Comparative fluorescein angiography of the normal sheep and goat ocular fundi

Fluorescein angiography without sedative or anesthetic agents was evaluated in 20 normal goats and 20 normal sheep. All of the angiographic phases were observed using 20 mg/kg fluorescein IV in both species. Fundus fluorescein angiography results revealed wide stars of Winslow in the tapetal fundus, central or marginal flow during the first part of the arterial phase, delayed filling of the focal areas in the choroid near the optic disc that often coincided with others in the disc, and lack of evidence of the 'striate area' in the tapetal fundi. In goats, the angiographic times were 6.54+/-1.25 s for the arterial phase (TA), 7.80+/-1.37 s for the arterio-venous phase (TAV), and 14.13+/-2.01 s for the venous phase (TV). I1: 1.30+/-0.30 s (time elapsing between TA and TAV), and I2: 6.20+/-1.60 s (time elapsing between TAV and TV). In sheep, times were 9.54+/-2.18 s TA, 11.73+/-2.10 s TAV, and 20.86+/-2.74 s TV. I1: 2.04+/-0.75 s and I2: 8.98+/-2.47 s, respectively. Due to the large size of the fundic vessels in sheep and goats, fluorescein angiography of the retinal vasculature can facilitate the study of the different vascular diseases in these species.     

Storage of spatial information by the maintenance mechanism of LTP

Analogous to learning and memory storage, long-term potentiation (LTP) is divided into induction and maintenance phases. Testing the hypothesis that the mechanism of LTP maintenance stores information requires reversing this mechanism in vivo and finding out whether long-term stored information is lost. This was not previously possible. Recently however, persistent phosphorylation by the atypical protein kinase C isoform, protein kinase Mzeta (PKMz), has been found to maintain late LTP in hippocampal slices. Here we show that a cell-permeable PKMz inhibitor, injected in the rat hippocampus, both reverses LTP maintenance in vivo and produces persistent loss of 1-day-old spatial information. Thus, the mechanism maintaining LTP sustains spatial memory.     

Functional markers delimiting a <Emphasis Type="Italic">Medicago</Emphasis> orthologue of pea symbiotic gene <Emphasis Type="Italic">NOD3</Emphasis>

Symbiotic gene mutated in the pea (Pisum sativum L.) line RisfixC is a determinant of the number of symbiotic root nodules. In parallel to a sharp increase in nodule number, its mutational inactivation brings about the insensitivity of nodulation to the ambient nitrate level (Nts trait). Using the established localization to the SYM2-NOD3 region of the pea linkage group I, functional PCR markers were developed for the orthologous region on the chromosome 5 of the model species Medicago truncatula. Owing to the conservation of the binding regions of the designed primers, pea orthologues were successfully amplified with 60% of the primer pairs tested. When applied to a mapping pea population from the cross of the line RisfixC x Afghanistan L1268 (sym2), the new markers allowed to localize the supernodulation mutation within 2.5 cM confidence interval in the pea genome. The placement of the functional markers on the M. truncatula chromosome 5 confined the orthologous gene location to eight overlapping BACs spanning approximately 710 kbp (positions 37,755,678–38,467,472). The narrowed list of the annotated Medicago genes in combination with the published data on their symbiotic and nitrate regulation can be used for the candidate gene identification, together with the requirements imposed by the known function in nodule number initiation and nitrate sensing. In addition, the new markers are applicable for tracking the RisfixC allele in breeding programmes aimed at the improvement of symbiotic performance.     

Improving the sensitivity of the IBR-gE ELISA for testing IBR marker vaccinated cows from bulk milk

The low sensitivity of the IBR-gE ELISA compared to other diagnostic ELISA tests for IBR is a major disadvantage of IBR control programmes based on IBR marker vaccination. Therefore the IBR-gE ELISA is not generally recommended for testing pooled or bulk milk samples.The aim of this study was to determine the performance of a commercially available kit for concentrating and purifying antibodies in milk in order to improve the sensitivity of detecting IBR-gE antibody positive cows from pooled and bulk milk samples. A single IBR-gE positive cow is likely to remain undetected in a pool of 49 negative milk samples without concentration. By contrast, the bulk milk concentration procedure improved sensitivity from 5.4% to 75.7% in a positive herd. Milk samples with a high or moderate positive signal are more likely to be detected after pool milk concentration compared to weak positive samples. Whereas a follow up study involving a monthly testing of bulk milk samples from three marker vaccinated IBR-gE negative herds over a period of seven months yielded negative results each month, bulk milk from a herd containing <5% IBR-gE positive cows always detected positive after concentration. Although the milk concentration procedure had no impact on specificity, it significantly enhanced the sensitivity of the detection of IBR-gE positive milk in pooled and bulk milk samples. After further evaluation this procedure could allow a cost efficient and reliable method of monitoring IBR marker-vaccinated herds for IBR-gE antibodies.     

Familial narcolepsy in the Lipizzaner horse: a report of three fillies born to the same sire

The occurrence of sleep disorder in three half sibling Lipizzaner is described. Sleepiness, swaying, stumbling, carpal joints buckling and falling down onto the carpal joints had been present since early foal age in all of them. Clinical signs had gradually reduced since the age of 2 years in cases 1 and 3. Sleepiness was induced by going out from the stable in adulthood. A physostigmine test was performed in all three affected horses and produced positive results in cases 1 and 3. The result of the test in case 2 was unclear due to the almost continuous sleepiness of the foal. Hypocretin-1 concentration in the cerebrospinal fluid was established using a standardised radioimmunoassay in case 1 (317.85?pg/mL), case 2 (303.43?pg/mL) and five adult control horses (275.2?±?47.9 [SD] pg/mL) and was considered as normal in all horses. The sire of the affected horses has had 19 other registered offspring who did not show clinical signs of sleep disorder and also dams of all three cases produced healthy foals. Based on the demographic and clinical data together with the responses to the physostigmine challenges, the diagnosis of familial equine narcolepsy was made.     

Estimation of stem attributes using a combination of terrestrial and airborne laser scanning

Properties of individual trees can be estimated from airborne laser scanning (ALS) data provided that the scanning is dense enough and the positions of field-measured trees are available as training data. However, such detailed manual field measurements are laborious. This paper presents new methods to use terrestrial laser scanning (TLS) for automatic measurements of tree stems and to further link these ground measurements to ALS data analyzed at the single tree level. The methods have been validated in six 80 × 80 m field plots in spruce-dominated forest (lat. 58°N, long. 13°E). In a first step, individual tree stems were automatically detected from TLS data. The root mean square error (RMSE) for DBH was 38.0 mm (13.1 %), and the bias was 1.6 mm (0.5 %). In a second step, trees detected from the TLS data were automatically co-registered and linked with the corresponding trees detected from the ALS data. In a third step, tree level regression models were created for stem attributes derived from the TLS data using independent variables derived from trees detected from the ALS data. Leave-one-out cross-validation for one field plot at a time provided an RMSE for tree level ALS estimates trained with TLS data of 46.0 mm (15.4 %) for DBH, 9.4 dm (3.7 %) for tree height, and 197.4 dm³ (34.0 %) for stem volume, which was nearly as accurate as when data from manual field inventory were used for training.     

Differential gene expression and apoptosis markers in presymptomatic scrapie affected sheep

Bluetongue viruses (BTVs) could invade N-W Europe similar to BTV serotype 8 (BTV8/net06), since the source and route of introduction of this virus has not been solved. Therefore, the Dutch survey for Bluetongue by PCR testing was extended by further analysis of PCR positives to identify the involved BTV. In late August 2008, BTV was reported with 12 nucleotide differences in the S10 amplicon (S10 genotyping). This virus was identified as serotype 6, here named BTV6/net08. Promptly, serotype specific real-time PCR tests were developed for serotypes 1, 6, and 8 (S2 genotyping). Agreement was found between results by S10- and S2 genotyping. Further, BTV1 was identified by both S10- and S2 genotyping in one imported animal. After initial discovery of BTV6 in the Netherlands, animals from 18 holdings tested PCR positive for BTV6/net08 in 2008. Remarkably only one or two PCR positive animals per holding were found. Serum neutralization tests did not result in the discovery of more BTV6 infected animals. Retrospective studies indicated no evidence for infections by BTV6/net08 prior to the first discovery. Experimental infections with BTV6/net08 did not cause clinical disease in sheep, calves and cattle, except for a very short fever in some animals. This clearly showed that the vaccine-related BTV6/net08 is not virulent. BTV6/net08 was not found by passive and active surveys in the years after its discovery. Apparently, BTV6/net08 was not efficiently transmitted by endemic species of Culicoides in N-W Europe, and disappeared without the need of any control measure.     

Induction of heme oxygenase-1 by Macleaya cordata extract and its constituent sanguinarine in RAW264.7 cells

Macleaya cordata (plume poppy) is used in traditional Chinese medicine for its anti-inflammatory and antibacterial activities. In this study, we examined whether M. cordata extract and/or its major alkaloid constituents, protopine, allocryptopine, sanguinarine and chelerythrine activate the Nrf2 signalling pathway which regulates the expression of cytoprotective enzymes including heme oxygenase-1 (HO-1) and thioredoxin 1. In murine macrophage RAW264.7 cells, M. cordata extract increased both mRNA and protein levels of HO-1. Of the alkaloids examined, only sanguinarine appeared to be responsible for these effects. At the concentration of 2 μM, sanguinarine induced nuclear accumulation of Nrf2, increased the expression of Hmox1 gene encoding HO-1 and elevated HO-1 protein levels. Sanguinarine-induced Hmox1 mRNA expression was suppressed by SB203580, a pharmacologic inhibitor of p38 mitogen-activated protein kinases (p38 MAPKs). The upregulation of HO-1 in RAW264.7 cells by sanguinarine was, however, accompanied by decrease in cell viability. Nonetheless, sanguinarine at micromolar, non-cytotoxic concentrations elevated protein levels of HO-1 and thioredoxin 1 in primary cultures of human hepatocytes. We conclude that sanguinarine may, under appropriate conditions, increase the capacity of the enzymatic antioxidant defence system via activation of the p38 MAPK/Nrf2 pathway.