Sequence analysis of the membrane protein gene and nucleocapsid gene of porcine reproductive and respiratory syndrome virus isolated from a swine herd in Hungary

Porcine reproductive and respiratory syndrome virus (PRRSV) was isolated from blood samples taken at a pig farm in Hungary from pigs showing clinical signs of the disease. The virus (ABV 32) was identified as belonging to the European genotype by using type-specific monoclonal antibodies. This was confirmed by comparing the sequence of the membrane protein gene (ORF 6) and the nucleocapsid gene (ORF 7) with the American VR2332 and the European LV genotype reference strain, respectively. Analysis of the amino acid sequence of the ORF 6 and ORF 7 of ABV 32 revealed five amino acid changes in both ORFs when compared with LV, of which two changes in ORF 7 were only found in the Spanish isolates. Additionally, the ORF 7 sequence was compared with corresponding sequences of a total of 21 other European strains. Phylogenetic analysis using the PHYLIP package confirmed the close relationship between the Hungarian and the Spanish isolates. Of all the isolates analysed, ABV 32 and LV were the least related

Current epizootiological status of the Eastern European countries for Aujeszky's disease

In recent years, several comprehensive reviews have been published on the results of Aujeszky's disease (AD) control in the Western European countries. The epizootiological data concerning Aujeszky's disease in the Eastern European (EE) countries are summarised for the first time. A questionnaire was drawn up according to uniform criteria and sent to the Chief Veterinary Authorities of the following countries, requesting experts of the given special field to participate in completing the questionnaire: Albania, Byelorussia, Bulgaria, Croatia, Czech Republic, Estonia, Hungary, Latvia, Lithuania, Macedonian Republic, Moldavia, Poland, Romania, Russia, Slovak Republic, Slovenia, Ukraine. Three main topics appeared in the questionnaire: (1) distribution of the pig population and structure of the farms; (2) current status and achievements of Aujeszky's disease control; (3) status of the national Aujeszky's disease eradication programme. The distribution of the pig population in the EE has been mapped by country on the basis of the pig densities. In the Eastern European countries the majority (in average 70-80%) of the pig holdings are farrowing and finishing farms. Aujeszky's disease belongs to the notifiable diseases in all countries. The number of cases occurring per year is reported to the O.I.E. The principles of Aujeszky's disease control are regulated by decrees issued by the Veterinary Authorities. Countries that have become free from Aujeszky's disease: Czech Republic (free status in 1988) (3 960 139 pigs), Slovenia (in 1996) (600 000 pigs), Republic of Estonia (in 1994) (330 700 pigs) and Latvia (in 1997) (403 540 pigs). Except for Latvia, the use of vaccine is prohibited in these countries. Aujeszky's disease free status is checked on a yearly basis by SNT or by ELISA (in Slovenia 3000-5000 sera/year, in Czech Republic 196 241 sera/1998, in Republic of Estonia all breeding pigs and fattening pigs (sent for slaughter) are examined/ year). Control is based on the use of a gE-deleted vaccine in all countries. In certain countries (e.g. in Poland and Hungary) both locally developed gE deleted vaccine and gE deleted vaccines manufactured by Western European companies are being used, while in other countries (e.g. in Slovak Republic and Bulgaria) exclusively locally developed vaccines are used for vaccination. Vaccination programmes are usually adapted to the epizootiological status of individual farms. Except for Hungary, backyard stocks are not vaccinated. In Hungary the infected backyard stocks are vaccinated three times per year. In Hungary and in Slovak Republic, the Aujeszky's disease eradication is carried out in the framework of a nation-wide eradication programme. In Poland and in Croatia such a nation-wide eradication programme is expected to be launched in the near future. In other countries the conditions are not yet ripe for starting national eradication programmes. We have not received any data from Rumania, Lithuania, Byelorussia and the countries presently situated in a war zone (Yugoslavia, Macedonian Republic). The EE countries are characterised by different levels of Aujeszky's disease control. Some countries have already declared their Aujeszky's disease free status while others are yet in the progress of creating the conditions necessary for eradication. While in the countries bordering Western Europe the principal objective of AD control is eradication of the disease, in the other countries the main goal is to reduce the economic losses caused by AD ("peaceful coexistence" with the virus). The difference between these goals arises mainly from the fact that countries aspiring to join the EU are aware that Aujeszky's disease will soon become a discriminatory factor on the international pig market. The EE countries bordering countries of Western Europe have a good chance of being able to declare their AD-free status by the expected date of their accession to the EU (2002-2004). $In\ memoriam$ We just learnt that Professor Istvan Medevczky quietly passed on November 1999. We would like to honour the memory of Professor Medevczky who was an estimated teacher in the University of Veterinary Science in Budapest and a recognized scientist at the international level. He was in charge to coordinate the Aujeszky's disease programme in Hungary. He participated actively to this symposium where he had the first symptoms of his disease. We present our deepest sympathy to his wife and family

Vaccinia virus replication is independent of cellular HSP72 expression which is induced during virus infection

HSP72 is dramatically induced in the ovaries of vaccinia virus (VV)-infected mice and associates with VV proteins. In order to Investigate the role of HSP72 during vaccinia virus replication, we have constructed a recombinant vaccinia virus encoding the major inducible cellular HSP72 (VV-HSP72+) and examined the replication characteristics of this virus. VV-HSP72+ exhibited growth kinetics identical to and peak titers very similar to those of control viruses, both in vitro and in vivo. In particular, replication of VV-HSP72+ was identical to that of control viruses in the HSP72-negative cell line Y3.Ag.1.2.3, and overexpression of HSP72 had no effect on the virulence of VV infection in normal or immunocompromised mice. We conclude that while W infection results in the induction of the major inducible 72-kDa HSP, VV replication proceeds normally in the absence of this protein. It is unclear whether another celluar chaperone is required to facilitate virus replication in place of HSP72 in Y3.Ag.1.2.3 cells or whether HSP expression plays no role in virus replication, but is simply a component of the generalized stress response to virus infection

Evolution and Prospects for Intracranial Pharmacotherapy for Refractory Epilepsies: The Subdural Hybrid Neuroprosthesis

Intracranial pharmacotherapy is a novel strategy to treat drug refractory, localization-related epilepsies not amenable to resective surgery. The common feature of the method is the use of some type of antiepileptic drug (AED) delivery device placed inside the cranium to prevent or stop focal seizures. This distinguishes it from other nonconventional methods, such as intrathecal pharmacotherapy, electrical neurostimulation, gene therapy, cell transplantation, and local cooling. AED-delivery systems comprise drug releasing polymers and neuroprosthetic devices that can deliver AEDs into the brain via intraparenchymal, ventricular, or transmeningeal routes. One such device is the subdural Hybrid Neuroprosthesis (HNP), designed to deliver AEDs, such as muscimol, into the subdural/subarachnoid space overlaying neocortical epileptogenic zones, with electrophysiological feedback from the treated tissue. The idea of intracranial pharmacotherapy and HNP treatment for epilepsy originated from multiple sources, including the advent of implanted medical devices, safety data for intracranial electrodes and catheters, evidence for the seizure-controlling efficacy of intracerebral AEDs, and further understanding of the pathophysiology of focal epilepsy. Successful introduction of intracranial pharmacotherapy into clinical practice depends on how the intertwined scientific, engineering, clinical, neurosurgical and regulatory challenges will be met to produce an effective and commercially viable device

Self-division of giant vesicles driven by an internal enzymatic reaction

Self-division is one of the most common phenomena in living systems and one of the most important properties of life driven by internal mechanisms of cells. Design and engineering of synthetic cells from abiotic components can recreate a life-like function thus contributing to the understanding of the origin of life. Existing methods to induce the self-division of vesicles require external and non-autonomous triggers (temperature change and the addition of membrane precursors). Here we show that pH-responsive giant unilamellar vesicles on the micrometer scale can undergo self-division triggered by an internal autonomous chemical stimulus driven by an enzymatic (urea-urease) reaction coupled to a cross-membrane transport of the substrate, urea. The bilayer of the artificial cells is composed of a mixture of phospholipids (POPC, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine) and oleic acid molecules. The enzymatic reaction increases the pH in the lumen of the vesicles, which concomitantly changes the protonation state of the oleic acid in the inner leaflet of the bilayer causing the removal of the membrane building blocks into the lumen of the vesicles thus decreasing the inner membrane area with respect to the outer one. This process coupled to the osmotic stress (responsible for the volume loss of the vesicles) leads to the division of a mother vesicle into two smaller daughter vesicles. These two processes must act in synergy; none of them alone can induce the division. Overall, our self-dividing system represents a step forward in the design and engineering of a complex autonomous model of synthetic cells