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E A Palombo

Publications and source records attributed to E A Palombo.

At least 19 recordsLinked to original sources
PubMed2001

Genetic relatedness of VP1 genes of Australian and Taiwanese rotavirus isolates.

Gene 1 (which encodes the viral RNA-dependent RNA polymerase, VP1) of an atypical human reassortant rotavirus strain, E210 (serotype G2P1B), is unrelated to genes 1 of standard human rotaviruses. To ascertain the origin of this gene, we determined a partial sequence and found that it exhibited greatest identity to gene 1 of a Taiwanese isolate, TE83, which is representative of G2 strains that caused an epidemic of gastroenteritis in 1993. Limited sequence identity to genes 1 of standard human and animal viruses was observed. This was confirmed by phylogenetic analysis. However, hybridization analysis using an E210 gene 1-specific probe indicated that a related gene was found among other Australian G2 isolates and in a Japanese strain isolated in the 1970s.

Australia↗
PubMed2001

Antibacterial activity of traditional Australian medicinal plants.

Fifty-six ethanolic extracts of various parts of 39 plants used in traditional Australian Aboriginal medicine were investigated for their antibacterial activities against four Gram-positive (Bacillus cereus, Enterococcus faecalis, Staphylococcus aureus and Streptococcus pyogenes) and four Gram-negative (Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Salmonella typhimurium) bacterial species. In a plate-hole diffusion assay, 12 extracts inhibited the growth of one or more of the bacteria, with five extracts showing broad spectrum antibacterial activity against Gram-positive bacteria. B. cereus was the most susceptible bacterium, with all 12 extracts displaying activity against this organism. Extracts from the leaves of Eremophila species (Myoporaceae) were the most active, with Eremophila duttonii exhibiting the greatest activity (against Gram-positive bacteria). The antibacterial effects of E. duttonii were further investigated by time-course growth assays which showed that significant growth inhibition was observed in cultures incubated in the presence of the extract within 1 h for B. cereus, E. faecalis and S. aureus and 2 h for S. pyogenes.

Anti-Bacterial Agents↗
PubMed2001

Rotavirus encephalopathy: pathogenesis reviewed.

Two cases of rotavirus gastroenteritis associated with neurological involvement, one with encephalitis (defined by abnormal neurological signs, cerebrospinal fluid (CSF) pleocytosis and detection of rotavirus genomic nucleic acid in the CSF) and one with a non-inflammatory encephalopathy (defined by abnormal neurological signs, an entirely normal CSF and detection of rotavirus genomic nucleic acid in the CSF), are presented and used as a basis to review and explore potential pathogenetic mechanisms, including direct viral replication within neurons and indirect effects of the newly described rotavirus 'enterotoxin'.

Acyclovir↗
PubMed2001

Characterisation and phylogenetic analysis of the VP7 proteins of serotype G6 and G8 human rotaviruses.

Serotype G6 and G8 rotaviruses are rarely found in man and may have originated in animals. Human serotype G6 and G8 rotaviruses, isolated from hospitalised children at various locations in Australia, were characterised. Deduced amino acid sequences of the major neutralising antigen, V7, showed significant identity to the cognate proteins of prototype human and bovine G6 and G8 viruses, respectively, and the strains reacted with G6 and G8 serotype-specific neutralising monoclonal antibodies, respectively, in an enzyme immunoassay. The VP4 type was determined as P[14] for all strains tested. Phylogenetic analysis of these and other human and bovine VP7 sequences suggested that a single inter-species transmission event, possibly from cattle, may have led to the emergence of G6 viruses in man. In contrast, the exchange of genes between human and bovine G8 viruses may have occurred onmore than one occasion, or these genes may have originated in a different host.

Amino Acid Sequence↗
PubMed2000

Detection of astrovirus gastroenteritis in children.

A commercial enzyme immunoassay (EIA) for the detection of astrovirus antigen was used to detect the virus during a 12-month survey of enteric pathogens in children in outpatient (n = 238) and hospital (n = 176) settings. It was found to have a 100% sensitivity and 98.6% specificity. Nineteen astrovirus isolates were detected and confirmed by northern hybridization, cell culture, and RT-PCR. The virus was detected mainly amongst outpatients although a comparison of the detection rate with that in hospitalised children did not demonstrate a statistically significant difference (p = 0.1347). In contrast, there was a strong association between hospitalization and rotavirus infection (p = 0.0371), and a strong association between infection detected in outpatients and adenovirus infection (p = 0.0193). Strains of astrovirus were sequenced, genotyped and shown to be: type 1 (n = 11), type 3 (n = 1), and type 4 (n = 7). Maximum genetic variation in type 1 isolates was 8.6% and type 4 was 7.8%. Changes did not result in amino acid substitutions.

Astroviridae Infections↗
PubMed2000

Epidemiology of astrovirus infection in young children hospitalized with acute gastroenteritis in Melbourne, Australia, over a period of four consecutive years, 1995 to 1998.

The incidence of astrovirus infection in children less than 5 years of age hospitalized with acute gastroenteritis in Melbourne, Australia, from 1995 to 1998 was determined. Astrovirus was detected in 40 of 449 specimens tested by Northern hybridization, and astrovirus infection was confirmed by reverse transcription-PCR with or without culture in CaCO-2 cells. This represented 3.0% (40 of 1, 327) of all children tested for enteric pathogens, including viral, bacterial, and parasitic pathogens, over the survey period. The incidences of astrovirus infection in each year were 4.4% (1995), 2. 2% (1996), 3.9% (1997), and 1.4% (1998). In 1995 and 1997, the incidences of astrovirus infection were greater than the incidence of infection with all individual bacterial pathogens and were either greater than or equal to the incidence of adenovirus infection. Astrovirus exhibited an unusual biennial winter peak of incidence that correlated with a greater incidence of serotype 1 virus and an increased rate of hospitalization of children aged 6 to 12 months. Uncommon (serotype 2 and 4) and rare (serotype 8) serotypes were detected during the survey period. Genetic analysis of ORF2 (which encodes the astrovirus capsid precursor) of Melbourne isolates showed nucleotide sequence variation from year to year. This was not accompanied by significant amino acid substitutions. However, geographical variation was apparent by comparison of Melbourne astrovirus isolates with prototype strains identified in the United Kingdom.

Acute Disease↗
PubMed2000

Molecular characterization of porcine rotaviruses from the southern region of Brazil: characterization of an atypical genotype G[9] strain.

The G (VP7) and P (VP4) serotype distribution of Brazilian porcine rotaviruses was determined using reverse transcription-PCR genotyping methods. Common porcine G types G3, G4, and G5 were detected in combination with P types [6] and [7]. The detection of nonporcine G types and unusual G-P combinations and the characterization of an atypical virus indicated that interspecies transmission may contribute to the genetic diversity of porcine rotaviruses.

Animals↗
PubMed2000

Characterisation of a "European-like" serotype G8 human rotavirus isolated in Australia.

An atypical human rotavirus strain, DG8, was isolated from a 13-month-old child hospitalised with acute gastro-enteritis in Australia. The virus could not be serotyped by enzyme immunoassay (EIA) using standard reagents specific for common Group A human rotavirus G serotypes. The deduced amino acid sequence of the outer capsid glycoprotein, VP7, indicated that this strain belonged to the uncommon human serotype G8. This was confirmed by EIA incorporating a G8-specific neutralising monoclonal antibody (NMAb). The VP4 genotype of DG8 was determined as P[14], equivalent to P serotype P3B, by sequence analysis and confirmed by EIA incorporating a P3B-specific NMAb. Electrophoretic analysis of DG8 genomic dsRNA indicated that the virus exhibited a "long" electropherotype. Northern hybridisation analysis (using a whole-genome probe derived from DG8) indicated that DG8 shared overall homology with the European serotype G8 strain, HAL1166 (11 of 11 genes). In contrast, only 9 of 11 genes of DG8 hybridised with the Asian serotype G8 strain, B37, and with the bovine G8 strain, A5. Hence, DG8 displayed features reminiscent of the human serotype G8 rotaviruses isolated in Europe in the mid-1980s rather than the geographically local G8 Asian strains isolated a decade earlier. It is possible that DG8 arose through reassortment between human and bovine rotaviruses.

Amino Acid Sequence↗
PubMed1999

Genetic and antigenic diversity of human rotaviruses: potential impact on the success of candidate vaccines.

Licensing of the first human rotavirus vaccine raises the hope of a reduction in the burden of paediatric diarrheal disease. However, the less than optimal performance of this vaccine in trials carried out in developing countries indicates that improvements in vaccine design are necessary. Analysis of the genetic and antigenic diversity of rotavirus isolates collected from various geographical locations suggests that future vaccine formulations may need to include a broader spectrum of strains. This may increase vaccine efficacy by providing comprehensive coverage against circulating viruses. Continued surveillance and genetic analysis of the rotavirus population prior to and after the introduction of routine vaccination will reveal if the diversity of this pathogen will impact on the success of vaccine programs.

Antigenic Variation↗
PubMed1999

Genetic and antigenic variation of capsid protein VP7 of serotype G1 human rotavirus isolates.

The deduced amino acid sequences of the outer capsid protein, VP7, of serotype G1 rotavirus clinical isolates collected over a 6 year period (1990-1995) in Melbourne, Australia, were examined. Phylogenetic analysis characterized the sequences into two discrete clusters representing two of the four global lineages of human G1 VP7 proteins. Antigenic characterization using a panel of serotype G1-specific neutralizing monoclonal antibodies classified lineage II isolates (1990-1993) as monotype G1a while lineage I isolates were classified as monotype G1b (1993-1995). Examination of the sequences of the neutralization epitope regions of VP7 revealed a particular amino acid substitution at residue 94 in region A (Asp --> Ser/Thr) that correlated with lineage and monotype designation. Our results indicated that temporal genetic variation of the VP7 of serotype G1 rotaviruses was associated with changes in the antigenicity of these isolates.

Amino Acid Sequence↗
PubMed1999

Genetic homogeneity of human serotype G1 rotaviruses isolated during a single epidemic season: implications for vaccine strategies.

The extent of genetic variation of rotavirus isolates representing the common serotype G1 circulating in Australian urban centres during 1996 was investigated. The sequences of the major outer capsid glycoprotein, VP7, of three isolates from Brisbane, Sydney and Melbourne were determined and found to be highly conserved (> 99% nucleotide identity, > 98.3% amino acid identity and > 99% amino acid similarity). In particular, the sequences of the neutralisation epitope regions were absolutely conserved. These strains and those from other cities were analysed by Northern blot hybridisation using a probe derived from a prototype serotype G1 virus. For all strains, the eleven genomic RNA segments hybridised with the probe indicating that these strains were not derived from genetic reassortment between different rotavirus geno-groups. These results suggest that, within a single epidemic season, rotavirus strains circulating in geographically diverse communities share common genetic features.

Amino Acid Sequence↗
PubMed1998

Improved sensitivity of astrovirus-specific RT-PCR following culture of stool samples in CaCo-2 cells.

BACKGROUND: During an epidemiological study on the incidence of astrovirus infection in children hospitalized with acute gastroenteritis, a Northern hybridization method was used to screen stool samples for astrovirus RNA. Positive results were confirmed using reverse transcriptase-polymerase chain reaction (RT-PCR), which showed surprisingly low sensitivity. The low sensitivity of the RT-PCR method was considered likely to be due to the presence of non-specific inhibitors. OBJECTIVE: To develop and use a simple culture method to improve the sensitivity of diagnosis of astrovirus in clinical stool samples using RT-PCR. STUDY DESIGN: Stool samples from children hospitalized with acute gastroenteritis were screened for astrovirus using Northern hybridization. The presence of astrovirus RNA was then confirmed using an astrovirus-specific RT-PCR. Hybridization positive samples that failed to generate an RT-PCR product were cultured in CaCO-2 cells for 48 h. RNA was isolated from cultures and re-tested using the same RT-PCR method. RESULTS: Using Northern hybridization, human astroviruses were detected in the stools of 31 patients and confirmed by RT-PCR in 16 samples. RNA extracted directly from 15 faecal specimens could not be amplified by RT-PCR. After culture for 48 h in CaCO-2 cells, RNA extracted from these samples could be amplified and confirmed the presence of astrovirus in all 15 specimens. CONCLUSIONS: Development of a simplified culture method for astrovirus positive faecal specimens improved the sensitivity of astrovirus-specific RT-PCR from 52 to 100%. The technique should be of value as a confirmatory test in surveys of human astrovirus infection.

Astroviridae Infections↗
PubMed1998

Sequence comparison of the VP7 of serotype G2 rotaviruses from diverse geographical locations.

The sequences of the genes encoding the outer capsid glycoprotein, VP7, of twelve serotype G2 rotavirus isolates from Australia were determined. The deduced amino acid sequences were compared by phylogenetic analysis to published sequences from strains collected from various geographical locations worldwide between 1976 an 1993. This analysis showed that geographical clustering of strains was apparent and that most strains were distantly related to strain DS1 whose VP7 is included in the rhesus rotavirus tetravalent vaccine formulation. Comparisons of the neutralization epitope regions of VP7 also indicated that most strains might exhibit antigenic differences to DS1. These results have implications for the development of future vaccine strategies.

Acute Disease↗
PubMed1998

Characterisation of rearranged NSP5 gene of a human rotavirus.

An atypical human rotavirus strain Z10262, isolated from a chronically infected immunodeficient child, displayed an unusual genomic RNA electrophoretic pattern. Besides, Northern blot analysis indicated that this strain contained an abnormally migrating gene 11 equivalent. Sequencing of this gene showed that it was derived from a genetic rearrangement which involved a partial duplication of the open reading frame (ORF) encoding the non-structural protein NSP5. However, the duplicated region contained a deletion and several point mutations relative to the first copy of the ORF. Phylogenetic analysis of human and animal NSP5 amino acid sequences including Z10262 revealed two groups of human proteins related to different animal proteins. The isolation and analysis of Z10262 strain provides further evidence for the genetic complexity of naturally occurring human rotaviruses.

Base Sequence↗
PubMed1997

Genetic characterization of the rotavirus nonstructural protein, NSP4.

The rotavirus nonstructural protein NSP4 plays a role in viral assembly by acting as an intracellular receptor for single-shelled particles and assisting in the translocation of these particles across the endoplasmic reticulum. Recently, NSP4 has been implicated in rotavirus virulence and is thought to act as an enterotoxin which triggers chloride secretion by a calcium-dependent signal transduction pathway. Limited sequence analysis of NSP4 shows a well-conserved protein. To define the extent of sequence variation in the gene coding for NSP4, we have sequenced this gene from nine human rotavirus strains. These data and the analysis of additional human strains and various animal rotaviruses (bovine, simian, equine, and porcine) by Northern blot hybridization suggested that three NSP4 genotypes were present among rotavirus strains. A correlation between NSP4 genotype and VP6 subgroup was also implied. Two different NSP4 genes (which encoded distinct types of NSP4 proteins) were found among standard human rotaviruses and in strains circulating in the local community and these showed homology to cognate genes in some animal strains.

Amino Acid Sequence↗
PubMed1997

Comparison of enzyme immunoassay, PCR, and type-specific cDNA probe techniques for identification of group A rotavirus gene 4 types (P types).

This study was designed to evaluate three techniques most commonly used to identify the VP4 (P) types of human group A fecal rotaviruses. The techniques included PCR with nested primers and hybridization with PCR-generated probes (to determine the P genotypes). The results obtained by these genetic techniques were evaluated against those obtained by an enzyme immunoassay (EIA) incorporating neutralizing monoclonal antibodies (N-MAbs) reacting with three major human P serotypes (serotypes P1A, P1B, and P2A). The P types of the rotaviruses present in 102 fecal specimens were determined under code by each of the three assays. The specificity of each assay was evaluated against a "gold standard" putative P type (P serotype and genotype) deduced from knowledge of the VP7 (G) type and the origin of the fecal specimen. Overall comparison of the results showed respective sensitivities and specificities of 92 and 92% for reverse transcription-PCR, 80 and 99% for hybridization, and 73 and 91% for EIA with N-MAbs. The hybridization assay retained high sensitivity with specimens stored for > or = 10 years. Hybridization assays with nonradioactive probes are relatively inexpensive and are suited for use in developing countries. In summary, both genetic assays showed high sensitivities and specificities in assigning a P type to human fecal rotavirus strains. Further evaluation of the EIA with N-MAbs is required, together with incorporation of new N-MAbs for the detection of the additional P types detected in developing countries.

Antibodies, Monoclonal↗