Analysis of receptor binding by the channel-forming toxin aerolysin using surface plasmon resonance.

Aerolysin is a channel-forming bacterial toxin that binds to glycosylphosphatidylinositol (GPI) anchors on host cell-surface structures. The nature of the receptors and the location of the receptor-binding sites on the toxin molecule were investigated using surface plasmon resonance. Aerolysin bound to the GPI-anchored proteins Thy-1, variant surface glycoprotein, and contactin with similar rate constants and affinities. Enzymatic removal of N-linked sugars from Thy-1 did not affect toxin binding, indicating that these sugars are not involved in the high affinity interaction with aerolysin. Aerolysin is a bilobal protein, and both lobes were shown to be required for optimal binding. The large lobe by itself bound Thy-1 with an affinity that was at least 10-fold weaker than that of the whole toxin, whereas the small lobe bound the GPI-anchored protein at least 1000-fold more weakly than the intact toxin. Mutation analyses provided further evidence that both lobes were involved in GPI anchor binding, with certain single amino acid substitutions in either domain leading to reductions in affinity of as much as 100-fold. A variant with single amino acid substitutions in both lobes of the protein was completely unable to bind the receptor. The membrane protein glycophorin, which is heavily glycosylated but not GPI-anchored, bound weakly to immobilized proaerolysin, suggesting that interactions with cell-surface carbohydrate structures other than GPI anchors may partially mediate toxin binding to host cells.     

Fine needle cytology of palpable head and neck lesions: a comparison of sampling methods with and without suction

Patients attending the ear, nose and throat (ENT) department at St James's University Hospital, Leeds, UK, for evaluation of palpable head and neck lesions have a fine needle cytology (FNC) specimen taken and receive the result at the same out-patient visit. This study was designed to discover if there is a significant difference in the efficiency of the methods with and without suction. The method was chosen randomly on each occasion and the adequacy or otherwise of the specimen was determined taking into account the site and nature of the lesion and the total cellularity of the sample. The level of blood contamination was also compared by each method. When benign and malignant lesions from all sites were analysed together the method with suction produced a significantly higher number of adequate samples than the method without suction. The exception was in the case of samples from lymph node lesions measuring < 1 cm, where adequate specimens were only obtained without suction. The non-suction technique was particularly poor at sampling salivary gland lesions in the 1–1.5 cm category. There was no significant difference in the level of blood contamination between the two methods at any site. These results are at variance with most other similar studies and possible reasons for this are discussed.     

The effect of dry cow winter management system on feed intake, performance and estimated energy demand

This research compared three wood-chip out-wintering pad (OWP; an unsheltered OWP; a sheltered OWP (both with a concrete feed apron); and an unsheltered OWP with silage provided directly on top of the wood-chip bedding (self-feed OWP)) designs and cubicle housing with regard to dairy cow performance during the pre-partum period, and for 8 weeks post partum. Data were compared during 2 years. In Year 1, the unsheltered (space allowance = 12 m2 per cow) and sheltered (6 m2 per cow) OWPs were compared with cubicle housing (n = 49 cows per treatment). In Year 2, all three OWP designs (12 m2 per cow) were compared with cubicle housing (n = 24 cows per treatment, split into two replicates). Animals were dried off and assigned to treatment in the autumn, and remained there until calving in spring. Subsequently, they were managed at pasture during lactation. Outcome measures for analysis during the pre-partum period were feed intake, live weight, body condition score (BCS), heat production and heat loss, and post-partum were live weight, BCS, milk yield and milk composition. In Year 1, all cows had a similar live weight, but both pre-partum and at calving cows on the unsheltered OWP had a lower BCS than cows in cubicles (P < 0.05). However, in Year 2, there were no differences in either live weight or BCS. In Year 1, cows in the unsheltered OWP produced less heat than in cubicles (P < 0.05), but in Year 2, there was no treatment effect. In both years, cows in unsheltered OWPs lost more heat than cows in the sheltered OWP (P < 0.001). Treatment had no effect on milk composition either year. However, in Year 2, cows in the self-feed OWP had higher milk yields than the other treatments (P < 0.05). The lower BCS and heat production values in unsheltered treatments during Year 1 were probably because of higher rainfall and wind-speed values of that year. However, in both years, live weight in all treatments increased pre partum, and BCS did not decrease, indicating that unsheltered cows did not need to mobilise body reserves. Thus, OWPs could be a suitable pre-partum alternative to cubicle housing for dry dairy cows with regard to some aspects of dairy cow productive performance. However, further research should be carried out to investigate longer-term effects.     

Antibody to the E3 glycoprotein protects mice against lethal venezuelan equine encephalitis virus infection

Six monoclonal antibodies were isolated that exhibited specificity for a furin cleavage site deletion mutant (V3526) of Venezuelan equine encephalitis virus (VEEV). These antibodies comprise a single competition group and bound the E3 glycoprotein of VEEV subtype I viruses but failed to bind the E3 glycoprotein of other alphaviruses. These antibodies neutralized V3526 virus infectivity but did not neutralize the parental strain of Trinidad donkey (TrD) VEEV. However, the E3-specific antibodies did inhibit the production of virus from VEEV TrD-infected cells. In addition, passive immunization of mice demonstrated that antibody to the E3 glycoprotein provided protection against lethal VEEV TrD challenge. This is the first recognition of a protective epitope in the E3 glycoprotein. Furthermore, these results indicate that E3 plays a critical role late in the morphogenesis of progeny virus after E3 appears on the surfaces of infected cells.