Conserved eukaryotic fusogens can fuse viral envelopes to cells

Caenorhabditis elegans proteins AFF-1 and EFF-1 [C. elegans fusion family (CeFF) proteins] are essential for developmental cell-to-cell fusion and can merge insect cells. To study the structure and function of AFF-1, we constructed vesicular stomatitis virus (VSV) displaying AFF-1 on the viral envelope, substituting the native fusogen VSV glycoprotein. Electron microscopy and tomography revealed that AFF-1 formed distinct supercomplexes resembling pentameric and hexameric "flowers" on pseudoviruses. Viruses carrying AFF-1 infected mammalian cells only when CeFFs were on the target cell surface. Furthermore, we identified fusion family (FF) proteins within and beyond nematodes, and divergent members from the human parasitic nematode Trichinella spiralis and the chordate Branchiostoma floridae could also fuse mammalian cells. Thus, FF proteins are part of an ancient family of cellular fusogens that can promote fusion when expressed on a viral particle.     

Antinociceptive efficacy and plasma concentrations of transdermal buprenorphine in dogs

To assess the antinociceptive efficacy of transdermal (TD) buprenorphine (B) in dogs, a prospective, positive-controlled experimental study was performed in 10 healthy Beagles. In an open label crossover design, the dogs initially received intravenous B (IVB, 0.02 mg kg^?1) as a positive control, followed by TDB (52.5 μg h^?1) 4 months later. Blood was collected at regular intervals for determination of the plasma concentrations of B ([B]) and its metabolite norbuprenorphine. The antinociceptive efficacy was assessed using thermal and mechanical models of nociception. The peak concentration [B] was 1.54 ng mL^?1 (±1.98) 60 h after TDB application, although three dogs had no measurable [B] after TDB. Maximum thermal threshold (TT) was 52.6 °C (±0.48) at 1 h after IVB administration and 51.63 °C (±1.01) 72 h after TDB application. The significant increase in TT indicated that effective antinociception was achieved beyond 36 h after the application of TDB, lasting until patch removal. There was hysteresis between [B] and the antinociceptive effect.     

Constituents in evening primrose oil with radical scavenging,cyclooxygenase, and neutrophil elastase inhibitory activities

Cold-pressed, non-raffinated evening primrose oil was found to contain lipophilic radical scavengers. A highly enriched fraction of these compounds could be obtained from the oil by extraction with aqueous ethanol and subsequent liquid-liquid partitioning with petroleum. LC-DAD-MS analysis revealed that the fraction contained three aromatic compounds with identical UV and ESI-MS spectra. The compounds were isolated by RP-HPLC and their structures established by chemical and spectroscopic means as 3-O-trans-caffeoyl derivatives of betulinic, morolic, and oleanolic acid. The morolic acid derivative was a new compound. The three esters exhibited pronounced radical scavenging activity against the stable 2,2-diphenyl-1-picrylhydrazyl radical and were potent inhibitors of neutrophil elastase and cyclooxygenase-1 and -2 in vitro. Commercial samples of evening primrose oils contained only traces of these lipophilic antioxidants.     

Plasma amino acid concentrations in aggressive dogs

Following the hypothesis that metabolic screens may be useful tools in the diagnosis of canine aggression we have investigated the blood plasma amino acid levels of dogs which have been found aggressive (N = 10) against dogs or men in comparison to non-aggressive dogs (N = 10). In summary, the aggressive dogs showed elevated plasma concentrations of the neurophysiological active aromatic amino acids tryptophan (46/171 micromol/l, p < 0,001), tyrosine (38/67 micromol/l, p < 0.01) and histidine (74/91 micromol/l, p < 0.01) and lower lysine concentrations (175/151 micromol/l, p < 0.05), which seems to point to a stress situation of these dogs. The nitrogen metabolism is impaired in the urea-cycle in the conversion of ornithine (17/34 micromol/l, p < 0.01) to citrulline (64/47 micromol/l). Higher levels of branched chain amino acids, especially leucine (122/150 micromol/l, p < 0.01), mainly metabolized in muscles, and isoleucin (60/71 micromol/l, p < 0.05) show a high energy potential.The acidose-stimulator methionine (48/78 micromol/l, p < 0.01) proved elevated. The results show that the changed behavior in the aggressive dogs is also reflected in their free amino acid plasma concentrations, independent of the question whether these data are the cause or the result of the aggressivity.     

Development and validation of species-specific primers that provide a molecular diagnostic for virus-vector longidorid nematodes and related species in German viticulture

The nematode species Longidorus attenuatus, L. elongatus, L. macrosoma and Paralongidorus maximusare economically important pests to the viticulture industry due to their ability to vector two nepoviruses (Raspberry Ringspot Virus and Tomato Black Ring Virus) to grapevines. In Germany, these species occur in vineyard soil with other non-vector but morphologically similar longidorid species, L. helveticus, L.profundorum and L. sturhani. Species-specific primers were designed from ribosomal DNA for all seven species to facilitate taxonomic identification for non-specialists. Primers were assessed for their reliability by screening, where possible, a number of populations of each species. Furthermore, their selectivity and sensitivity were determined when challenged with closely related longidorid species and general nematode communities typical of vineyard soil. A multiplex approach using a common forward primer combined with species-specific reverse primers enabled three target nematode species to be detected in the same PCR reaction. All primers were highly specific, detecting all nematode developmental forms from disparate populations and were sufficiently sensitive to detect a single target nematode within a whole nematode community typical of a vineyard soil comprising of a range of non-target species. Given their specificity, sensitivity and reliability, these diagnostic primers should be of great benefit to both phytosanitary/quarantine services related to the viticulture industry and also as a decision management tool for growers.     

Phospholipid-based microemulsions suitable for use in foods

The preparation of nonaqueous microemulsions using food-acceptable components is reported. The effect of oil on the formation of microemulsions stabilized by lecithin (Epikuron 200) and containing propylene glycol as immiscible solvent was investigated. When the triglycerides were used as oil, three types of phase behavior were noted, namely, a two-phase cloudy region (occurring at low lecithin concentrations), a liquid crystalline (LC) phase (occurring at high surfactant and low oil concentrations), and a clear monophasic microemulsion region. The extent of this clear one-phase region was found to be dependent upon the molecular volume of the oil being solubilized. Large molecular volume oils, such as soybean and sunflower oils, produced a small microemulsion region, whereas the smallest molecular volume triglyceride, tributyrin, produced a large, clear monophasic region. Use of the ethyl ester, ethyl oleate, as oil produced a clear, monophasic region of a size comparable to that seen with tributyrin. Substitution of some of the propylene glycol with water greatly reduced the extent of the clear one-phase region and increased the extent of the liquid crystalline region. In contrast, ethanol enhanced the clear, monophasic region by decreasing the LC phase. Replacement of some of the lecithin with the micelle-forming nonionic surfactant Tween 80 to produce mixed lecithin/Tween 80 mixtures of weight ratios (Km) 1:2 and 1:3 did not significantly alter the phase behavior, although there was a marginal increase in the area of the two-phase, cloudy region of the phase diagram. The use of the lower phosphatidylcholine content lecithin, Epikuron 170, in place of Epikuron 200 resulted in a reduction in the LC region for all of the systems investigated. In conclusion, these studies show that it is possible to prepare one-phase, clear lecithin-based microemulsions over a wide range of compositions using components that are food-acceptable.     

Laryngeal mask airway insertion requires less propofol than endotracheal intubation in dogs

OBJECTIVE: To compare the doses of propofol required for insertion of the laryngeal mask airway (LMA) with those for endotracheal intubation in sedated dogs. STUDY DESIGN: Randomized prospective clinical study. Animals Sixty healthy dogs aged 0.33-8.5 (3.0 +/- 2.3, mean +/- SD) years, weighing 2.2-59.0 (23.4 +/- 13.6, mean +/- SD) kg, presented for elective surgery requiring inhalation anaesthesia. METHODS: Animals were randomly assigned to receive either a LMA or an endotracheal tube. Pre-anaesthetic medication was intravenous (IV) glycopyrrolate (0.01 mg kg(-1)) medetomidine (10 microg kg(-1)) and butorphanol (0.2 mg kg(-1)). Repeated IV propofol injections (1 mg kg(-1) in 30 seconds) were given until LMA insertion or endotracheal intubation was achieved, when the presence or absence of laryngospasm, the respiratory rate (fr) and the total dose of propofol used were recorded. RESULTS: The total propofol dose (mean +/- SD) required for LMA insertion (0.53 +/- 0.51 mg kg(-1)) was significantly lower than for endotracheal intubation (1.43 +/- 0.57 mg kg(-1)). The LMA could be inserted without propofol in 47% of dogs; the remainder needed a single 1 mg kg(-1) bolus (n = 30). Endotracheal intubation was possible without propofol in 3.3% of the dogs, 47% needed one bolus and 50% required two injections (n = 30). The f(r) (mean +/- SD) was 18 +/- 6 and 15 +/- 7 minute(-1) after LMA insertion and intubation, respectively. CONCLUSION AND CLINICAL RELEVANCE: Laryngeal mask airway insertion requires less propofol than endotracheal intubation in sedated dogs therefore propofol-induced cardiorespiratory depression is likely to be less severe. The LMA is well tolerated and offers a less invasive means of securing the upper airway.     

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.     

Measurement of tidal volume using Respiratory Ultrasonic Plethysmography in anaesthetized,mechanically ventilated horses

ObjectiveTo compare tidal volume estimations obtained from Respiratory Ultrasonic Plethysmography (RUP) with simultaneous spirometric measurements in anaesthetized, mechanically ventilated horses.Study designProspective randomized experimental study.AnimalsFive experimental horses.MethodsFive horses were anaesthetized twice (1 week apart) in random order in lateral and in dorsal recumbency. Nine ventilation modes (treatments) were scheduled in random order (each lasting 4 minutes) applying combinations of different tidal volumes (8, 10, 12 mL kg^?1) and positive end-expiratory pressures (PEEP) (0, 10, 20 cm H₂O). Baseline ventilation mode (tidal volume = 15 mL kg^?1, PEEP = 0 cm H₂O) was applied for 4 minutes between all treatments. Spirometry and RUP data were downloaded to personal computers. Linear regression analyses (RUP versus spirometric tidal volume) were performed using different subsets of data. Additonally RUP was calibrated against spirometry using a regression equation for all RUP signal values (thoracic, abdominal and combined) with all data collectively and also by an individually determined best regression equation (highest R²) for each experiment (horse versus recumbency) separately. Agreement between methods was assessed with Bland-Altman analyses.ResultsThe highest correlation of RUP and spirometric tidal volume (R² = 0.81) was found with the combined RUP signal in horses in lateral recumbency and ventilated without PEEP. The bias ± 2 SD was 0 ± 2.66 L when RUP was calibrated for collective data, but decreased to 0 ± 0.87 L when RUP was calibrated with individual data.Conclusions and clinical relevanceA possible use of RUP for tidal volume measurement during IPPV needs individual calibration to obtain limits of agreement within ± 20%.     

Effects of brine salting with regard to raw material variation of Atlantic salmon (Salmo salar) muscle investigated by Fourier transform infrared microspectroscopy

Atlantic salmon fillets differing with regard to raw material characteristics (prerigor, postrigor, frozen/thawed) and salt content were investigated by FT-IR microspectroscopy and light microscopy. Local variation within each salmon fillet was further taken into account by sampling from the head and tail part separately as they vary in fat and moisture content. The highest salt uptake was achieved for frozen/thawed quality during brine-salting with 16% NaCl for 4 h, while the uptake was least for prerigor fish. At the same time, salting caused muscle fiber swelling of about 10% for both frozen/thawed and postrigor qualities. Differences in the FT-IR amide I spectral region were observed implying a change in the muscle protein secondary structure. Prerigor was least affected by brine-salting, having a final salt concentration of 2.2%, while postrigor had a NaCl content of 3.0% and frozen/thawed of 4.1%. Local variation within the fillets had an effect on the amide I absorption characteristics before as well as after salting. Salt uptake of the samples was affected by raw material quality and at the same time the degree of swelling of the myofibers was influenced by raw material character.