Nucleotide Sequences of IRES Domains IV and V of Natural ECHO Virus type 11 Isolates with Different Replicative Capacity Phenotypes

ECHO viruses (ECV) belong to the enterovirus genus of the Picornaviridae family and are the most frequently isolated from clinical and environmental samples. They are responsible for a wide variety of clinical syndromes involving most organs of the human body. We previously postulated that some of the variations in the recognition of ECHO virus type 11 (ECV 11) strains by a group specific monoclonal antibody (Mab) which we have studied could be explained by variations in their replicative capacity in cell culture and variations within the 5′ nontranslated region (5′ NTR) of their genomes. To support this hypothesis, the replicative capacity in cell culture and the nucleotide sequences of domains IV and V of the IRES of the genome of five ECV11 strains (the Gregory reference strain and four wild isolates) were determined, and analysed. Our results indicate that the replicative capacity of wild ECV11 isolates studied by one-step growth cycle in both HEp-2 and Vero cell cultures showed variations among strains in comparison with the Gregory reference strain. The clinical ECV11 strains replicated as well as the reference strain, however environmental strains displayed a phenotype with a significant reduction of replication. The sequences of ECV 11 strains showed significant conservation with that of the poliovirus (PV1) Mahoney strain The comparative examination of the predicted secondary structures revealed, that the nucleotide variations did not affect the secondary structure of stem-loop structure IV and V in the IRES element, however differences were especially observed in the apical stem region (nucleotides 483 to 509) of the domain V of the ECV11 strains and resulted in modification of the central stem structure. The nucleotide sequences data reported in this paper have been submitted to the GenBank nucleotide sequence databases and have been assigned the accession No DQ029083 to DQ029096.     

Hepatitis A in Tunisia: phylogenetic analysis of hepatitis A virus from 2001 to 2004

Tunisia is a highly endemic area for hepatitis A virus (HAV) infection. In the present study, the phylogenetic characterization of the VP1 gene (882 nucleotides) and of the VP1/2A junction (336 nucleotides) of Tunisian strains were examined. One hundred strains isolated from patient with anti-HAV IgM from 2001 to 2004 were amplified by RT-PCR, sequenced at the VP1 and at the VP1/2A junction and aligned with the published sequences to establish phylogenetic analysis. All Tunisian strains belong to genotype I with a greater presence of sub-genotype IA (98%) originate from most of Tunisian regions and 2% of sub-genotype IB. In addition, sub-genotype IA and IB strains formed 25 different clusters. Genetically similar strains were also identified between 2001 and 2004 isolated from the southern and the central part of Tunisia, suggesting that an indigenous strain has been circulating in the Tunisia. The genetic profile of the VP1 region showed that Tun159-02 and Tun40-03 clustered respectively in the IB and IA sub-genotype, however, analysis of VP1/2A junction revealed in contrast that Tun159-02 and Tun40-03 clustered respectively in IA and IB. This is the first report to identify sub-genotype IA in Tunisia and provides new data on the genetic relatedness of HAV from Tunisia and the distribution of sub-genotype IA in this part of the world.     

Pernicious anemia in a young man with systemic lupus erythematosus

Association of pernicious anemia and systemic lupus erythematosus is rare, although both diseases are autoimmune origin. We describe the case of a 40-year old man with systemic lupus erythematosus (SLE) who developed pernicious anemia. The cobalamin deficiency was revealed by macrocytic pancytopenia. After 1 month of vitamin B12 treatment, hemoglobin and white blood cell count remain normal but thrombocytopenia persists and was considered as immunologic from SLE origin requiring corticosteroids.     

Study of in vitro cytotoxic activity of cyclopentanones derivatives

The cytotoxicity of 30 cyclopentanone derivatives was studied in vitro, in a clonogenic assay using murine leukemia L1210. Results are compared to those obtained with reference medicaments. 17 derivatives exhibit activities against L1210 cells.     

Evaluation of an enterovirus group-specific anti-VP1 monoclonal antibody, 5-D8/1, in comparison with neutralization and PCR for rapid identification of enteroviruses in cell culture.

We evaluated the usefulness of a commercially available monoclonal antibody (MAb) directed against a group-specific epitope of the capsid protein VP1 of enteroviruses for the rapid identification of these viruses in cell culture. The MAb was assayed in an indirect immunofluorescence test with cultured cells infected by various serotypes of enterovirus; all 39 serotypes tested, including echoviruses 22 and 23, which are considered atypical enteroviruses, were reactive. The MAb was also tested with 61 strains recovered from clinical specimens inoculated into cell cultures in comparison with seroneutralization with intersecting pools of hyperimmune sera and PCR with primers from the 5' untranslated region of enteroviruses. There was total agreement between the results obtained with the MAb and those obtained by PCR, even for those strains of enteroviruses which were found to be untypeable with polyclonal antisera. These data demonstrate the usefulness of the MAb for rapid identification of enteroviruses in cell culture.     

Neuromeningeal enterovirus infections in Tunisia: epidemiology, clinical presentation, and outcome of 26 pediatric cases

Non-polio enteroviruses are the most common identified cause of viral neuromeningeal infections following the introduction of the mumps and polio vaccines. OBJECTIVE: The aim of this study was to describe the epidemiology, clinical presentation, and the outcome of enteroviral infections of the CNS. METHOD AND PATIENTS: We performed a prospective study on 41 children admitted for viral neuromeningeal infections in the pediatric department of Monastir between December 2001 and November 2002. Enteroviruses were detected from cerebrospinal fluid by RT-PCR. RESULTS: This study showed that enteroviruses were responsible for 63.4% of the infections. The mean age of patients was 6.1 years. Aseptic meningitis was diagnosed in 14 cases and encephalitis in 10. The most frequent symptom was fever (61.5%), followed by seizures (42.3%), and confusion (23%). On follow-up, all patients with meningitis had recovered without sequels. Neurological complications in patients with encephalitis were epilepsy (3 cases), cerebral palsy (2 cases), and mental retardation (1 case). CONCLUSION: This study confirmed that enteroviruses were the most common cause of viral infections of the CNS. Common use of RT-PCR can have a significant impact on the outcome of patients with enterovirus infections.