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Biomedical subjects

L Haas

Publications and source records attributed to L Haas.

At least 37 records · Page 2Linked to original sources
PubMed1999

Rapid and sensitive detection of immunoglobulin M (IgM) and IgG antibodies against canine distemper virus by a new recombinant nucleocapsid protein-based enzyme-linked immunosorbent assay.

Canine distemper morbillivirus (CDV) infection causes a frequently fatal systemic disease in a broad range of carnivore species, including domestic dogs. In CDV infection, classical serology provides data of diagnostic and prognostic values (kinetics of seroconversion) and is also used to predict the optimal vaccination age of pups. Routine CDV serology is still based on time- and cost-intensive virus neutralization assays (V-NA). Here, we describe a new capture-sandwich enzyme-linked immunosorbent assay (ELISA) that uses recombinant baculovirus-expressed nucleocapsid (N) protein of a recent CDV wild-type isolate (2544/Han95) for the detection of CDV-specific antibodies in canine sera. Recombinant antigen was produced with high efficacy in Heliothis virescens larvae. The capture-sandwich ELISA enabled a clear-cut qualitative evaluation of the CDV-specific immunoglobulin G (IgG) and IgM serostatuses of 196 and 35 dog sera, respectively. Inter-rater agreement analysis (kappa = 0.988) indicated that the ELISA can be used unrestrictedly as a substitute for the V-NA for the qualitative determination of CDV-specific IgG serostatus. In an attempt to semiquantify N-specific antibodies, a one-step-dilution (alpha method) IgG-specific ELISA was implemented. Alpha values of >/=50% showed very good inter-rater agreement (kappa = 0.968) with V-NA titers of >/=1/100 50% neutralizing dose (ND50) as measured against the central European CDV wild-type isolate 2544/Han95 in canine sera originating from northern Germany. An ND50 titer of 1/100 is considered a threshold, and titers of >/=1/100 indicate a resilient, protective immunity. CDV N-specific antibodies of the IgM class were detected by the newly developed ELISA in 9 of 15 sera obtained from dogs with symptoms of acute distemper. In leucocytes of 5 of the 15 dogs (all of which were also IgM positive) CDV RNA was detected by reverse transcription (RT)-PCR. The recombinant capture-sandwich ELISA detecting N-specific antibodies of the IgG class provided superior sensitivity and specificity and thus represents a rapid and cost-effective alternative to classical CDV V-NA. By detection of specific IgM antibodies, the ELISA will be complementary to RT-PCR and V-NA in the diagnosis of acute distemper infections.

Animals↗
PubMed1999

What could be causing chronic abdominal pain? Anything from common peptic ulcers to uncommon pancreatic trauma.

Establishing a reasonable diagnosis and treatment plan in patients with chronic abdominal pain can be difficult and frustrating. Most cases involve a common and readily identified condition (e.g., gallbladder disease, GERD, irritable bowel syndrome). Other cases, however, resist ready diagnosis because they offer no intra-abdominal explanation. In this article, the authors summarize several diverse possibilities as the source of the pain, and they describe how to approach evaluation to avoid unnecessary testing.

Abdominal Injuries↗
PubMed1999

[Emergence of "new" viral zoonoses].

In the last two to three decades a significant increase of viral zoonotic infections was observed. These zoonoses are not only newly (or previously unrecognized) emerging diseases, but also due to the reappearance of diseases thought to have been defeated (re-emerging diseases). "New" viral diseases can arise when viruses broaden their host-range (monkey poxvirus; equine morbillivirus), or can be a consequence of intrinsic properties of the virus itself, such as high mutation rates (influenza A virus). Most new or reemerging viral zoonoses are due to infections with hemorrhagic viruses. Many of them are transmitted by insects (arboviruses, e.g. yellow fever virus) or by rodents (e.g. Hanta viruses), others by contact with patients and nosocomial infections (e.g. Ebola virus). The emergence and increase of these diseases are a consequence of anthropogenic environmental changes, such as distortions of the ecological balance and changes in agriculture. In addition, the uncontrolled growth of the cities in tropical and subtropical regions without improvement of the public health measures and the increasing international animal trade and travel also favour the spread and recurrence of these diseases.

Animals↗
PubMed1998

Application of a computer program for genetic typing of classical swine fever virus isolates from Germany.

The commercial software program HLA SequiTyper (Amersham Pharmacia Biotech), designed originally for human leukocyte antigen typing, was adapted for rapid typing of classical swine fever (CSF) virus isolates. The program compares new sequence data with those stored in a database file and calculates the most probable assignment. For generating the CSF virus sequence database, 150 bp of the 5' nontranslated genomic region (5'-NTR) from 88 German classical swine fever virus isolates from outbreaks between 1984 and 1997 were solid-phase sequenced directly after RT-PCR amplification. Sequence alignments showed that they all belonged to the previously defined genetic group 2. Within this group, six different subgroups could be distinguished, and were designated according to the geographic location where they are either still endemic or where they appeared most commonly. The advantage of using the HLA SequiTyper program is that it reads directly the sequence files as generated by the ALF sequencer (Amersham Pharmacia Biotech), making any manipulations unnecessary. In addition, a constant quality control of the raw sequence data can be achieved, as more than one sequence from the same isolate can be evaluated at once. Using this approach, new CSF isolates can be typed within 2 days.

Animals↗
PubMed1998

Genetic analysis of the central untranslated genome region and the proximal coding part of the F gene of wild-type and vaccine canine distemper morbilliviruses.

Located between the open reading frames encoding the matrix (M) and the fusion (F) protein the morbillivirus genome contains an unusually large non-coding intercistronic region (M-F UTR) of up to 5.6% of the full length genome. Any function(s) of this region have largely remained obscure. Here, we analyze the M-F UTR and the proximal coding part of the downstream F gene of several recent canine distemper morbillivirus (CDV) wild-type (wt) isolates and vaccine strains. While the F gene coding part appeared to be highly conserved (about 93% homology), a considerable degree of strain-specific variation of up to 21.4% was evident when comparing the M-F UTR. Phylogenetic analysis revealed a co-circulation of several contemporary CDV genotypes within a close geographic range (central Europe). A remarkably distinct CDV wt lineage, so far detected only in mustelids, is displayed. A rather non-scattered pattern of mutations within the M-F UTR suggested superimposition of RNA sequence and/or secondary structure constraints. Extensive folding in the long (460 nt) and moderately GC-rich 5'-UTR of the F mRNA was evident, particularly around the putative F protein translation initiation codon (AUG461 of the Onderstepoort vaccine strain). The region immediately preceding the putative F initiation site also harbored the only mutation unique to both vaccine strains within the F-5'UTR (position 455: Awt vs. Cvac). The putative F protein start codon, AUG461, was found to be mutated to AUA or GUA in all wt isolates analyzed and in another vaccine strain (Rockborn). Possible consequences for F protein translation initiation in wt CDV are discussed.

Animals↗
PubMed1998

Cultures in conflict: a challenge to faculty of academic health centers.

Academic health centers (AHCs) are experiencing turmoil in all three of their traditional missions of teaching, research, and patient care. The authors examine origins of universities and medical education to place in historical context the stresses affecting AHCs at the end of the 20th century. They describe the cultures of the university to suggest strategies for successful adaptation to these stresses. Clashes of values and norms of the cultures within universities and AHCs can hinder effective adaptation to external change. Administrators, researchers, teachers, and clinicians can have strongly conflicting perspectives. For example, business skill is of increasing importance to the survival of the clinical enterprise, but not typically valued by faculty members. University faculty have often considered accountability as antithetical to academic freedom, and, until recently, accountability was not strongly demanded of AHCs. The authors conclude that AHC faculty must transcend the outdated view that the roles of the scholar, scientist, and healer are in opposition to those of the leader and manager. If AHCs are to survive and prosper through their current cultural transition, their faculty must understand all these roles as part of their intellectual and organizational responsibility.

Academic Medical Centers↗
PubMed1997

The development of early vs. late onset mucosal disease is a consequence of two different pathogenic mechanisms.

Bovine viral diarrhea (BVD) virus is the causative agent of fatal mucosal disease (MD) of cattle. Experimental induction of MD can be achieved by superinfection of calves persistently viremic with a noncytopathic (ncp) BVD virus using an antigenically similar cytopathic (cp) BVD virus. Here we describe the characterisation of BVD viruses isolated from three cases of experimentally induced MD. One animal developed clinical symptoms two weeks after superinfection (early onset MD), while the onset of disease in the other two cases occurred with a delay of months (late onset MD). Antigenic characterisation of the viruses was performed using a panel of monoclonal antibodies against the E2 glycoprotein. For genetic analysis, RT-PCR was applied to amplify specific insertions and duplications in the NS2-3 genomic region of the cp BVD viruses. In addition, these amplicons and fragments of the viral E2 genes were sequenced. The results showed that in the case of early onset MD the cp BVD virus isolated after begin of disease was identical to the one used for superinfection. In contrast, the cp BVD viruses isolated from the two animals with late onset MD were obviously the result of genetic recombinations between the persistent ncp and the superinfecting cp BVD viruses. We conclude that early and late onset MD are the consequence of different pathogenic mechanisms.

Amino Acid Sequence↗
PubMed1997

Analysis of the haemagglutinin gene of current wild-type canine distemper virus isolates from Germany.

The haemagglutinin (H) gene sequences from three wild-type canine distemper viruses (CDV) isolated during 1994-1995 were sequenced to determine whether contemporary strains had undergone significant genetic changes relative to the currently used vaccine strains. The new isolates were closely related to each other (> 99%) and displayed about 90-91% sequence homology to the Onderstepoort and Convac vaccine strains. There were one to four additional potential glycosylation sites compared to the vaccine strains which were also present in a German dog CDV isolate dating from 1990. However, only a very slight reduction in neutralizing titre against the new isolates was found when compared with the Onderstepoort and Rockborn vaccine strains. Cysteine and proline residues were well conserved indicating a conserved three dimensional structure for the protein. By phylogenetic analysis the recent isolates showed a narrow clustering close to the previous canine isolates indicating a linear pattern of evolutionary changes. A comparison with published CDV H gene sequences suggested the presence of different lineages of CDV on a global scale and possible cocirculation of more than one genotype of CDV.

Amino Acid Sequence↗
PubMed1997

Molecular epidemiology of animal virus diseases.

In this review the application of methods of molecular epidemiology, particularly the combined approach of amplifying defined fragments of viral genomes using the polymerase chain reaction and subsequent nucleotide sequencing analysis, is described. Emphasis is put on examples of a few important diseases (e.g. those caused by morbilliviruses, rhabdoviruses, pestiviruses and aphthoviruses) to demonstrate the impact of this methodology. Molecular epidemiology is already an important and very sensitive tool to study the evolution of viruses at a level superior to previous methodologies and providing a better understanding of epidemiological relationships.

Animal Diseases↗
PubMed1997

Characterization of the genome of feline foamy virus and its proteins shows distinct features different from those of primate spumaviruses.

The genome of the feline foamy virus (FeFV) isolate FUV was characterized by molecular cloning and nucleotide sequence analysis of subgenomic proviral DNA. The overall genetic organization of FeFV and protein sequence comparisons of different FeFV genes with their counterparts from other known foamy viruses confirm that FeFV is a complex foamy virus. However, significant differences exist when FeFV is compared with primate foamy viruses. The FeFV Gag protein is smaller than that of the primate spumaviruses, mainly due to additional MA/CA sequences characteristic of the primate viruses only. Gag protein sequence motifs of the NC domain of primate foamy viruses assumed to be involved in genome encapsidation are not conserved in FeFV. FeFV Gag and Pol proteins were detected with monospecific antisera directed against Gag and Pol domains of the human foamy virus and with antisera from naturally infected cats. Proteolytic processing of the FeFV Gag precursor was incomplete, whereas more efficient proteolytic cleavage of the pre125Pro-Pol protein was observed. The active center of the FeFV protease contains a Gln that replaces an invariant Gly residue at this position in other retroviral proteases. Functional studies on FeFV gene expression directed by the promoter of the long terminal repeat showed that FeFV gene expression was strongly activated by the Bell/Tas transactivator protein. The FeFV Bell/Tas transactivator is about one-third smaller than its counterpart of primate spumaviruses. This difference is also reflected by a limited sequence similarity and only a moderate conservation of structural motifs of the different foamy virus transactivators analyzed.

Amino Acid Sequence↗
PubMed1996

Canine distemper virus infection in Serengeti spotted hyenas.

Clinical signs suggestive of canine distemper virus (CDV) infection were observed among a group of spotted hyenas (Crocuta crocuta) in the Serengeti, Tanzania. Virus antigen was detected immunohistologically in a brain sample from a diseased cub. The presence of virus RNA could be demonstrated in this brain as well as in intestine and lymph node of the animal by RT-PCR. Sequence comparison of brain-derived amplicons showed that the virus was related to recent CDV field isolates. The closest homology (>99 percent) was to a recently described CDV which caused high mortality in sympatric lions.

Animals↗
PubMed1996

Rinderpest and other animal morbillivirus infections: comparative aspects and recent developments.

The genus morbillivirus presently comprises measles virus of man, rinderpest virus (RPV), peste des petits ruminants virus (PPRV), and canine distemper virus (CDV). 'Emerging' morbilliviruses, such as phocid distemper virus (PDV) of seals, dolphin (DMV) and porpoise morbillivirus (PMV) have probably been present for a long period of time and outbreaks are possibly related to introduction into a highly susceptible population and/or be the result of interspecies transmission. In this review some comparative aspects of morbillivirus infections, particularly with respect to rinderpest and canine distemper viruses, are presented. Topics include pathogenesis, epidemiology, molecular phylogeny, diagnosis and prophylaxis. Recent developments in molecular biology have created tools which have enabled us to achieve a better understanding of morbillivirus infections at the nucleic acid level ('molecular epidemiology') while recombinant DNA technology has allowed new bivalent recombinant vaccines with improved heat stability to be produced.

Animals↗
PubMed1996

A case report of paroxysmal dystonic choreoathetosis due to hypoglycaemia induced by an insulinoma.

Hypoglycaemia due to an insulinoma can mimic acute disorders of cognition, consciousness, epilepsy, transient ischaemia, or psychosis, and chronic disorders of dementia and neuropathy. Misdiagnosis and delay in treatment are common and prolonged hypoglycaemia can lead to permanent neurological deficit or fatal coma. A 27 year old woman with hypoglycaemia induced by an insulinoma presented with features typical of paroxysmal non-kinesiogenic dystonic choreoathetosis. Striatal dysfunction as a consequence of hypoglycaemia has rarely been described. Insulinoma is a readily treatable condition that should be considered in the differential diagnosis of a paroxysmal movement disorder.

Adult↗
PubMed1995

Editing of morbillivirus P gene transcripts in infected animals.

RNA editing in the Morbillivirus genus in vivo was investigated by applying a polymerase chain reaction-based primer extension technique to measure the edited and non-edited mRNA transcripts. In this genus of the Paramyxoviridae the P gene transcript is altered by the co-transcriptional addition of one extra G residue to produce the mRNA for the V non-structural protein. Using tissues of phocine distemper virus (PDV) infected seals, canine distemper virus (CDV) infected dogs and rinderpest virus (RPV) infected cattle, it was demonstrated that editing occurs in vivo. The P:V mRNA ratios were generally similar to those found in tissue culture infections with the same virus and a minor fraction of transcripts had 2-4 extra G residues. In one seal brain infected with PDV the ratio of P:V mRNA was reversed but no differences were found in the levels of mRNA editing in different tissues from the same animal infected with CDV or RPV. However, variation was seen between animals infected with different isolates of RPV and even between animals infected with the same isolate of RPV.

Animals↗
PubMed1995

Experimentally induced "late-onset" mucosal disease--characterization of the cytopathogenic viruses isolated.

Antigenic and genetic analyses were performed in order to establish relationships between the noncytopathogenic (ncp) and the cytopathogenic (cp) bovine viral diarrhoea viruses (BVDV) involved in the induction of a case of experimentally induced "late-onset" mucosal disease (MD) symptoms. The persistent ncpBVDV, the cpBVDV used for superinfection (strain TGAC) and the virus isolates from faeces (cpX) were examined using an immunoplaque test (IPT) to distinguish between cp and ncp virus populations. The cp populations were cloned by plaque purification and found to be free of ncpBVDV when using the IPT. The cpBVDV clones and the persistent ncpBVDV were analysed in an enzyme immunoassay on heat-fixed infected cells (IM-EIA) and in a neutralization test using a panel of 27 monoclonal antibodies against the E0 (gp48) and E2 (gp53) viral glycoproteins. It was found that strain TGAC contained two antigenically distinct subpopulations of cpBVDV (TGAC-B1 and TGAC-B2). The endogenous ncpBVDV and the cpX clones had the same reactivity pattern in both tests. In addition, p80 gene duplications in the genomes of the cpBVDV clones were analysed using the polymerase chain reaction and subsequent restriction enzyme analysis of the amplicons. The clones analysed from TGAC-B1 and those from cpX had gene duplications of identical sizes showing the same restriction enzyme patterns. Our results suggest that the cpBVDV which finally lead to "late-onset" MD arose by recombination and/or by mutations of the cpBVDV used for superinfection.

Animals↗
PubMed1995

Serial MRI in listeria mesenrhombencephalitis: a case report.

Listeria mesenrhombencephalitis is a rare disorder, especially in the non-immunocompromised host. The predilection for brain stem involvement is unexplained. The anatomical correlation of brain stem signs with serial MRI over three months is shown in a 64 year old white man. The pathological lesions seemed to be a combination of micro-abscesses and associated oedema.

Encephalitis↗
PubMed1993

Engraftment of bovine leukemia virus infected lymphocytes into SCID mice.

SCID mice inoculated with peripheral blood lymphocytes from a tumor-bearing BLV-positive cow developed lymphosarcomas after 6 months. DNA from the splenic tumor specifically hybridized with a bovine Sstl satellite probe, showing that these tumor cells were of bovine origin. The clonality of the SCID splenic tumor, as measured by the integration pattern of BLV proviruses, was the same as that in the donor lymphocyte population.

Animals↗