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The genome of the swine influenza H1N1 virus Sweden/83 was compared to other H1N1 viruses by oligonucleotide fingerprinting. This analysis revealed that Sweden/83 is identical to the Danish strain Sjaelland/82 and is thus to be placed in the group of 'US viruses'. On the other hand, the Danish strain Als/82 was found to be different, i.e., related to the Belgium/79 strain which is a representative of the 'European viruses'. The results of the present genomic analysis confirm the previous findings of relationships between the above strains. Zusammenfassung Eine Analyse einiger H1N1-Viren mittels Oligonukleotid 'Fingerprint' Das Genom des Schweine-Influenzavirus H1N1 Schweden/83 wurde mit anderen H1N1-Viren mittels Oligonukleotid 'Fingerprint' verglichen. Diese Analyse zeigte, daß der Virustamm Schweden/83 mit dem dänischen Stamm Sjaelland/82 identisch ist und daher zu der US-Virusgruppe gehört. Es wurde jedoch festgestellt, daß der dänische Stamm Als/82 sich von den anderen unterscheidet und mit dem Stamm Belgium/79 eng verwandt ist, der zu der Gruppe europäischer Viren gehört. Die vorliegenden Ergebnisse der Genomanalyse bestätigen frühere Feststellungen einer Verwandtschaft zwischen den obengenannten Stämmen. [ABSTRACT FROM AUTHOR]

Direct filter hybridization (DFH) was evaluated as a rapid detection system for the diagnosis of Aujeszky's disease (AD) in pigs. Infection was detected in nasal and tonsillar cells, lymphocytes and in organ specimens by filter hybridization without previous purification of nucleic acids. The results of DFH agreed with those obtained by virus isolation and conventional nucleic acid hybridization. The infection was demonstrated by DFH both in acutely diseased and in clinically recovered animals. The sensitivity and specificity of both isotope-labelled and biotinylated probes were compared and confirmed. Zusammenfassung Einfacher Nachweis der Aujeszkyschen Krankheit bei Schweinen mittels DNS-Hybridisierung Für die Diagnostik der Aujeszkyschen Krankheit bei Schweinen wurde der Nachweis von virusspezifischer DNS durch direkte Filter-Hybridisierung (DFH) entwickelt. Dabei konnte eine Infektion in Nasen- und Tonsillentupfern, Lymphozyten und Organproben nachgewiesen werden. Die Ergebnisse lassen sich bestätigen durch Isolierung des Virus und die bisher übliche DNS-Hybridisierung. Sensitivität und Spezifität wurden für radioaktiv und Biotinmarkierte Sonden verglichen. [ABSTRACT FROM AUTHOR]

Swine influenza was first diagnosed in Sweden in April of 1983 when outbreaks occurred in the south of the country and three H1N1 strains were isolated from diseased pigs on three farms. The strains were closely related to each other and so one of them was chosen for comparison with six reference H1N1 strains from abroad. The Swedish strain proved to be closely related to one of two strains isolated during an epidemic of swine influenza in Denmark in 1981-1984. This Danish strain (Sjaelland) and the Swedish strain were more closely related to the NJ/76 strain than to any of the other strains tested. The comparison was made by serology and analysis of migration patterns of viral polypeptides and viral RNA segments. Zusammenfassung Der Ursprung des schwedischen Schweine-Influenzavirus H1N1 aus deḿ Jahre 1983 Schweineinfluenza wurde in Schweden erstmals im April 1983 diagnostiziert, als im Süden des Landes Ausbrüche auftraten. Drei H1N1-Stämme wurden aus erkrankten Tieren dreier Betriebe isoliert. Die Stämme wiesen untereinander eine enge Beziehung auf, so daß einer von ihnen zum Vergleich mit sechs ausländischen H1N1-Referenzstämmen ausgewählt wurde. Der schwedische Stamm zeigte sich als eng verwandt mit einem von zwei Stämmen, die in Dänemark während einer Schweineinfluenza-Epidemie in den Jahren 1981-1984 isoliert worden waren. Dieser dänische Stamm (Sjaelland) und der schwedische waren enger verwandt mit dem NJ/76-Stamm, als mit irgendeinem anderen der getesteten. Der Vergleich wurde serologisch durchgeführt sowie durch Analyse der Wanderungsmuster viraler Polypeptide und RNA-Segmente. [ABSTRACT FROM AUTHOR]

The DNA assisted solid-phase proximity ligation assay (SP-PLA) provides a unique opportunity to specifically detect prion protein (PrP) aggregates by investigating the collocation of 3 or more copies of the specific protein. We have developed an SP-PLA that can detect PrP aggregates in brain homogenates from infected hamsters even after a 10(7)-fold dilution. In contrast, brain homogenate from uninfected animals did not generate a detectable signal at 100-fold higher concentration. Using either of the 2 monoclonal anti-PrP antibodies, 3F4 and 6H4, we successfully detected low concentrations of aggregated PrP. The presented results provide a proof of concept that this method might be an interesting tool in the development of diagnostic approaches of prion diseases.

The prion protein (PrP) sequence of European moose, reindeer, roe deer and fallow deer in Scandinavia has high homology to the PrP sequence of North American cervids. Variants in the European moose PrP sequence were found at amino acid position 109 as K or Q. The 109Q variant is unique in the PrP sequence of vertebrates. During the 1980s a wasting syndrome in Swedish moose, Moose Wasting Syndrome (MWS), was described. SNP analysis demonstrated a difference in the observed genotype proportions of the heterozygous Q/K and homozygous Q/Q variants in the MWS animals compared with the healthy animals. In MWS moose the allele frequencies for 109K and 109Q were 0.73 and 0.27, respectively, and for healthy animals 0.69 and 0.31. Both alleles were seen as heterozygotes and homozygotes. In reindeer, PrP sequence variation was demonstrated at codon 176 as D or N and codon 225 as S or Y. The PrP sequences in roe deer and fallow deer were identical with published GenBank sequences.

Aim: To evaluate weekly induction chemotherapy followed by weekly concomitant chemoradiotherapy in a multicentre phase II study of patients with unresectable stage III non-small cell lung cancer (NSCLC; stage wet IIIB excluded)., Patients and Methods: Eligible patients received three weekly cycles of paclitaxel 100 mg/m2 and carboplatin AUC2 followed by six weekly cycles of paclitaxel 60 mg/m2 and carboplatin AUC2 in combination with thoracic radiotherapy (2 Gy per fraction and day to a total dose of 60 Gy)., Results: Sixty-four patients (40 males and 24 females) with a median age of 63 years (range, 43-79 years) entered the study. T and N stage were distributed as follows: T1 2 patients (3.2%), T2 10 patients (15.6%), T3 15 patients (23.4%), T4 37 patients (57.8%); N0 10 patients (15.6%), N1 1 patient (1.6%), N2 26 patients (40.6%), N3 26 patients (40.6%), and N missing 1 patient (1.6%). Seven patients (10.9%) suffered from grade 3/4 oesophagitis. Grade 1/2 oesophagitis occurred in 36 patients (56.3%) and pneumonitis grade 1/2 occurred in 10 patients (15.6%). Sixty-three patients were evaluated on an intent-to-treat basis. The overall response rate was 74.6%. The median time to progression was 247 days and median overall survival was 461 days. According to subgroup analyses, no statistically significant differences were noted according to gender, age (<65 vs. > or =65 years), performance status, histology, or study centre., Conclusion: Induction chemotherapy followed by concurrent chemoradiotherapy with weekly cycles of paclitaxel and carboplatin is feasible and generates moderate toxicity. Efficacy is comparable to other recently published regimens. However, prognosis remains, in general, poor for this group of patients and further work to develop better therapy is required.

This European Community Biomedicine and Health Research-supported, multicenter, randomized, placebo-controlled, double-blind trial investigated the effect of an angiotensin-converting enzyme inhibitor (ACE-I) in children and young people with IgA nephropathy (IgAN), moderate proteinuria (>1 and <3.5 g/d per 1.73 m(2)) and creatinine clearance (CrCl) >50 ml/min per 1.73 m(2). Sixty-six patients who were 20.5 yr of age (range 9 to 35 yr), were randomly assigned to Benazepril 0.2 mg/kg per d (ACE-I) or placebo and were followed for a median of 38 mo. The primary outcome was the progression of kidney disease, defined as >30% decrease of CrCl; secondary outcomes were (1) a composite end point of >30% decrease of CrCl or worsening of proteinuria until > or =3.5 g/d per 1.73 m(2) and (2) proteinuria partial remission (<0.5 g/d per 1.73 m(2)) or total remission (<160 mg/d per 1.73 m(2)) for >6 mo. Analysis was by intention to treat. A single patient (3.1%) in the ACE-I group and five (14.7%) in the placebo group showed a worsening of CrCl >30%. The composite end point of >30% decrease of CrCl or worsening of proteinuria until nephrotic range was reached by one (3.1%) of 32 patients in the ACE-I group, and nine (26.5%) of 34 in the placebo group; the difference was significant (log-rank P = 0.035). A stable, partial remission of proteinuria was observed in 13 (40.6%) of 32 patients in the ACE-I group versus three (8.8%) of 34 in the placebo group (log-rank P = 0.033), with total remission in 12.5% of ACE-I-treated patients and in none in the placebo group (log-rank P = 0.029). The multivariate Cox analysis showed that treatment with ACE-I was the independent predictor of prognosis; no influence on the composite end point was found for gender, age, baseline CrCl, systolic or diastolic BP, mean arterial pressure, or proteinuria.

An increasing number of scrapie cases with atypical characteristics, designated Nor98, have recently been recognized. Here, the proteinase K (PK)-resistant prion protein (PrP) fragments from two Swedish cases of Nor98 atypical scrapie have been characterized. The prominent, fast-migrating band in the distinct Nor98 Western immunoblot electrophoretic profile was determined to be of 7 kDa in size and was accordingly designated Nor98-PrP7. The antigenic composition of Nor98-PrP7, as assayed by a panel of anti-PrP antibodies, revealed that this fragment comprised a mid-region of PrP from around aa 85 to 148. N- and C-terminally truncated fragments spanning the mid-region of PrP have only been observed in the genetic prion disorder Gerstmann-Sträussler-Scheinker disease. It is shown here that the long-term PK resistance of Nor98-PrP7 is reduced compared with that of PrP(res) in classical scrapie. Enzymic deglycosylation did not change the distinct electrophoretic profile of Nor98-PrP7. A previously unidentified, PK-resistant, C-terminal PrP fragment of around 24 kDa was detected and its PK resistance was investigated. After deglycosylation, this fragment migrated as a 14 kDa polypeptide and was designated PrP-CTF14. Antigenic determination and the size of 14 kDa suggested a fragment spanning approximately aa 120-233. The existence of two PK-resistant PrP fragments, Nor98-PrP7 and PrP-CTF14, that share an overlapping region suggests that at least two distinct PrP conformers with different PK-resistant cores are present in brain extracts from Nor98-affected sheep. The structural gene of PrP in three Nor98-affected sheep was analysed, but no mutations were found that could be correlated to the aberrant PK-resistant profile observed.

We have compared the processing, turnover and release of bovine PrP (boPrP) in transfected baby hamster kidney (BHK) and mouse neuroblastoma (N2a) cells. In BHK cells, boPrP was subjected to two distinct proteolytic cleavage events, the first was mapped between K(121) and H(122) generating an N-terminal and a C-terminal PrP fragment. Transport block experiments, cell surface biotinylation and PIPLC analyses showed that the bulk of boPrP on the cell surface was the C-terminal fragment and indicated that the first cleavage of boPrP took place prior to or very soon after it appears at the cell surface. The second cleavage was situated at the extreme C-terminus of the boPrP GPI-anchored C-terminal fragment and as a result of this was shed into the medium rapidly. The kinetics, the migration in SDS-PAGE of the released fragment and protease inhibition studies indicate that a proteolytic activity was responsible for the release of the boPrP fragment from its GPI-anchor. Both N- and C-terminal fragments of boPrP could be detected in the medium. Moreover, in normal bovine brain, a C-terminal fragment was identified, suggesting that similar proteolytic processing events occur in vivo. In N2a cells, the majority of boPrP was subjected to a more complete degradation process, and only trace amounts of full length boPrP was shed into cell culture medium in a process which also indicated a release by proteolytic cleavage.

In this paper we show that the porcine rubulavirus LPMV phosphoprotein (P) and V protein (V) both interact with the nucleoprotein (NP). There are also indications for an interaction between P and V with L protein. Further analysis of the domains of the P and V which are necessary for interaction with the NP protein demonstrates that the interaction is not mediated from their common part but instead from their unique C-terminal parts, respectively. The common N-terminus of P and V appear to mediate the interaction with L. We also map the regions of NP that are necessary for interaction with P and V, respectively. Both P and V interact with regions of NP, which reside in the N-terminal part but appear not to overlap.

In order to study persistence of the porcine rubulavirus LPMV, we examined tissue samples collected from pigs 53 days after experimental infection. These pigs survived the initial infection and could clinically be considered to have recovered from the infection. Two of the pigs used in this study were chemically immunosuppressed during the last 4 days before necropsy. No infectious virus or viral antigen could be detected in any tissue using standard methods for virus isolation and detection. However, the presence of viral genomic RNA and mRNA could be demonstrated in the mid brain of the convalescent pig using an optimised RT-nested PCR. Mid brain, forebrain and lung were all shown to contain LPMV RNA in the immunosuppressed convalescent pigs. In addition we examined the P-gene editing in the recovered pigs and conclude that the viral genome is transcriptionally active in these pigs. The relevance of the persistence of LPMV for maintenance and spread within and/or between pig populations is discussed.