cDNA-cloning and expression of VP1-specific sequences of foot-and-mouth disease virus types A5 and O1

The RNA genome of foot- and mouth disease virus strains A5 Westerwald and O1 Lausanne has been reverse-transcribed and cloned in lambdaphages or plasmids. Identification of cDNA-clones containing VP1-specific sequences was achieved by hybridization, restriction mapping, and sequence analysis. VP1-coding cDNA-fragments were subcloned into the expression vector pEX which led to synthesis of fusion proteins with beta-galactosidase. These fusion proteins reacted with anti-VP1 antibodies on a Western blot, but were not capable of inducing neutralizing antibodies to mice. This seemed to suggest a tertiary structure of the VP1-epitopes unlike those of native VP1. Other attempts are discussed to construct VP1-fusion proteins folding similarly to the native viral protein structure.     

Modulation of nociceptive withdrawal reflexes evoked by single and repeated nociceptive stimuli in conscious dogs by low-dose acepromazine

ObjectivesTo investigate the modulation of the nociceptive withdrawal reflex (NWR) and temporal summation (TS) by low‐dose acepromazine (ACP) in conscious dogs. To assess the short‐ and long‐term stability of the reflex thresholds.Study designRandomized, blinded, placebo‐controlled cross‐over experimental study.AnimalsEight adult male Beagles.MethodsThe NWR was elicited using single transcutaneous electrical stimulation of the ulnar nerve. Repeated stimuli (10 pulses, 5 Hz) were applied to evoke TS. The responses of the deltoideus muscle were recorded and quantified by surface electromyography and the behavioural reactions were scored. Each dog received 0.01 mg kg^?1 ACP or an equal volume saline intravenously (IV) at 1 week intervals. Measurements were performed before (baseline) and 20, 60 and 100 minutes after drug administration. Sedation was scored before drug administration and then at 10 minutes intervals. Data were analyzed with Friedman repeated measures analysis of variance on ranks and Wilcoxon signed rank tests.ResultsAcepromazine resulted in a mild tranquilization becoming obvious at 20 minutes and peaking 30 minutes after injection. Single (I_t) and repeated stimuli (TS_t) threshold intensities, NWR and TS characteristics and behavioural responses were not affected by the ACP at any time point. Both I_t and TS_t were stable over time.Conclusions and clinical relevanceIn dogs, 0.01 mg kg^?1 ACP IV had no modulatory action on the NWR evoked by single or repeated stimuli, suggesting no antinociceptive activity on phasic nociceptive stimuli. The evidence of the stability of the NWR thresholds supports the use of the model as an objective tool to investigate nociception in conscious dogs. A low dose of ACP administered as the sole drug, can be used to facilitate the recordings in anxious subjects without altering the validity of this model.     

Real-time observation of molecular motion on a surface

The laser-induced movement of CO molecules over a platinum surface was followed in real time by means of ultrafast vibrational spectroscopy. Because the CO molecules bound on different surface sites exhibit different C-O stretch vibrational frequencies, the site-to-site hopping, triggered by excitation with a laser pulse, can be determined from subpicosecond changes in the vibrational spectra. The unexpectedly fast motion--characterized by a 500-femtosecond time constant--reveals that a rotational motion of the CO molecules, rather than pure translation, is required for this diffusion process. This conclusion is corroborated by density functional theory calculations.     

High-power directional emission from microlasers with chaotic resonators

High-power and highly directional semiconductor microcylinder lasers based on an optical resonator with deformed cross section are reported. In the favorable directions of the far-field, a power increase of up to three orders of magnitude over the conventional circularly symmetric lasers was obtained. A "bow-tie"-shaped resonance is responsible for the improved performance of the lasers in the higher range of deformations, in contrast to "whispering-gallery"-type modes of circular and weakly deformed lasers. This resonator design, although demonstrated here in midinfrared quantum-cascade lasers, should be applicable to any laser based on semiconductors or other high-refractive index materials.     

Structural dynamics of eukaryotic chromosome evolution

Large-scale genome sequencing is providing a comprehensive view of the complex evolutionary forces that have shaped the structure of eukaryotic chromosomes. Comparative sequence analyses reveal patterns of apparently random rearrangement interspersed with regions of extraordinarily rapid, localized genome evolution. Numerous subtle rearrangements near centromeres, telomeres, duplications, and interspersed repeats suggest hotspots for eukaryotic chromosome evolution. This localized chromosomal instability may play a role in rapidly evolving lineage-specific gene families and in fostering large-scale changes in gene order. Computational algorithms that take into account these dynamic forces along with traditional models of chromosomal rearrangement show promise for reconstructing the natural history of eukaryotic chromosomes.     

Local Control of Microdomain Orientation in Diblock Copolymer Thin Films with Electric Fields

Local control of the domain orientation in diblock copolymer thin films can be obtained by the application of electric fields on micrometer-length scales. Thin films of an asymmetric polystyrene-polymethylmethacrylate diblock copolymer, with cylindrical polymethylmethacrylate microdomains, were spin-coated onto substrates previously patterned with planar electrodes. The substrates, 100-nanometer-thick silicon nitride membranes, allow direct observation of the electrodes and the copolymer domain structure by transmission electron microscopy. The cylinders aligned parallel to the electric field lines for fields exceeding 30 kilovolts per centimeter, after annealing at 250°C in an inert atmosphere for 24 hours. This technique could find application in nanostructure fabrication.     

Purification of secretory immunoglobulin A from milk of the brushtail possum (Trichosurus vulpecula)

AIM: To identify and purify secretory immunoglobulin A (sIgA), a key effecter molecule in mucosal immune responses, from milk of the brushtail possum (Trichosurus vulpecula).

METHODS: Milk samples were collected from female possums with pouch young, and clarified by centrifugation and precipitation methods. The clarified fraction was purified by gel filtration and affinity chromatography to yield sIgA. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting techniques were used to assess the purity of the final product, and to identify the heavy (H) chain, light (L) chain and secretory component (SC) of possum sIgA.

RESULTS: Immunoblotting, using antibodies raised against cloned possum sIgA SC and H-chain, and a synthetic peptide fragment of the H-chain, confirmed the identity of the purified protein. The N-terminal amino acid sequence of purified possum sIgA showed strong homology to reported sequences of H-chain variable regions of marsupial immunoglobulins.

CONCLUSIONS: Milk was shown to be a convenient source of mucosal secretion containing sIgA, and a process involving 2 precipitation and 2 chromatography steps produced purified sIgA. This IgA preparation will prove useful for the generation of sIgA-specific immunological reagents for measurement of immune responses in the development of mucosal-based vaccines for biological control of possums.     

Unilateral Seminoma in a Dromedary Camel

A 10‐year‐old, clinically healthy, male dromedary camel had presented a history of progressive unilateral testicular enlargement over the past 5 years. The animal had mated with 32 females during that period; all had conceived. The sex ratio of his offspring was one male to 31 females. Ultrasound examination of the right testicle revealed a diffusely heterogeneous parenchyma with no identifiable normal testicular tissue. The enlarged testicle was surgically removed. Macroscopically, the testicle had a glistening pink surface and contained multiple soft, bulging nodules. Histopathologically, a well‐differentiated, diffuse seminoma was diagnosed. In conclusion, this study describes the fertility, sex ratio, clinical findings and ultrasonographic imaging in a male dromedary camel affected with unilateral testicular seminoma.