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Biomedical subjects

J Curran

Publications and source records attributed to J Curran.

At least 73 records · Page 4Linked to original sources
PubMed1992

Complexes of Sendai virus NP-P and P-L proteins are required for defective interfering particle genome replication in vitro.

We present evidence that the formation of NP-P and P-L protein complexes is essential for replication of the genome of Sendai defective interfering (DI-H) virus in vitro, using extracts of cells expressing these viral proteins from plasmids. Optimal replication of DI-H nucleocapsid RNA required extracts of cells transfected with critical amounts and ratios of each of the plasmids and was three- to fivefold better than replication with a control extract prepared from a natural virus infection. Extracts in which NP and P proteins were coexpressed supported replication of the genome of purified DI-H virus which contained endogenous polymerase proteins, but extracts in which NP and P were expressed separately and then mixed were inactive. Similarly, the P and L proteins must be coexpressed for biological activity. The replication data thus suggest that two protein complexes, NP-P and P-L, are required for nucleocapsid RNA replication and that these complexes must form during or soon after synthesis of the proteins. Biochemical evidence in support of the formation of each complex includes coimmunoprecipitation of both proteins of each complex with an antibody specific for one component and cosedimentation of the subunits of each complex. We propose that the P-L complex serves as the RNA polymerase and NP-P is required for encapsidation of newly synthesized RNA.

Animals↗
PubMed1991

The Sendai virus P gene expresses both an essential protein and an inhibitor of RNA synthesis by shuffling modules via mRNA editing.

The P gene of Sendai virus expresses as many as eight proteins, two of which (V and W) are expressed only from edited mRNAs; only the P protein is known to be involved in RNA synthesis. To examine the functions of the other P gene proteins, we developed an in vivo system in which genome replication is driven by plasmid generated viral proteins. We found that P was essential for this process, whereas V and W were not only non-essential, they were inhibitory. By using various P gene deletions and varying the amounts of plasmids transfected, we provide evidence that P is a modular protein. The N-terminal domain (shared with V and W) binds the L or polymerase protein, whereas the C-terminal domain binds the nucleoprotein NP. A model of paramyxovirus RNA synthesis is presented, and the implications of negative regulation during persistent infection are discussed.

Blotting, Northern↗
PubMed1991

A retrospective study of myocardial infarction occurring in elderly patients in a coronary care unit.

Myocardial infarction is a major cause of morbidity and mortality particularly in the elderly. Coronary Care Unit monitoring has resulted in the improvement in prognosis in all age groups. In this study, the clinical course and outcome of 204 consecutive patients with acute myocardial infarction aged over 65 years is retrospectively analysed. All patients were managed in a Coronary Care Unit operating a needs related policy. Those under 75 years (120 patients) were compared to those over that age (84 patients). The groups differed in terms of clinical presentation, cardiac enzymatic rise and response to drug therapy. However they had similar complication rates, morbidity and mortality.

Age Factors↗
PubMed1991

Characterization of the Sendai virus V protein with an anti-peptide antiserum.

The Sendai virus V protein, which is a fusion of the P and V ORFs of the P gene, was characterized with antisera to a portion of the V ORF and compared to the P protein. The only property found in common with P is that V is also highly phosphorylated, and this is so even when these proteins are expressed independently of the other viral proteins. Otherwise, V was not found in virions, was not strongly associated with viral nucleocapsids like P, and anti-V had no effect on viral RNA synthesis in vitro under conditions where anti-P was highly inhibitory. The available evidence suggests that V may play a role in RNA synthesis, but it is not an essential one like that of the P protein.

Amino Acid Sequence↗
PubMed1991

The P gene of human parainfluenza virus type 1 encodes P and C proteins but not a cysteine-rich V protein.

The nucleotide sequence of the P gene of human parainfluenza virus type 1 (PIV1) was determined from cloned cDNA copies of the mRNA. By analogy with the gene organization of Sendai virus, two open reading frames in the mRNA sense of the gene were identified as coding sequences for the P protein (568 amino acids with an estimated molecular weight of 64,655) and the C protein (204 amino acids with an estimated molecular weight of 24,108). Comparison of the deduced amino acid sequences of the P and C proteins of PIV1 with those of Sendai virus showed a high degree of homology. However, a sequence for the cysteine-rich V protein, which was considered a common feature of other paramyxoviruses, was interrupted by the presence of multiple stop codons. The sequence analysis of three P-gene-specific cDNA clones generated from genomic RNA by polymerase chain reaction and one additional clone generated from mRNA confirmed that the coding sequence for the cysteine-rich region is silent in the PIV1 gene and thus is not translated into protein. Two potential editing sites with the consensus sequence 3'UUYUCCC were found in the PIV1 P gene at positions 564 to 570 and 1430 to 1436. However, examination of the PIV1 mRNA population by a primer extension method indicated that neither of these sites is utilized. These results indicate that the PIV1 P gene has a coding strategy different from those of other paramyxovirus P genes.

Amino Acid Sequence↗
PubMed1991

The P gene of bovine parainfluenza virus 3 expresses all three reading frames from a single mRNA editing site.

The P gene of bovine parainfluenza virus 3 (bPIV3) contains two downstream overlapping ORFs, called V and D. By comparison with the mRNA editing sites of other paramyxoviruses, two editing sites were predicted for bPIV3; site a to express the D protein, and site b to express the V protein. Examination of the bPIV3 mRNAs, however, indicates that site b is non-functional whereas site a operates frequently. Insertions at site a give rise to both V and D protein mRNAs, because a very broad distribution of Gs is added when insertions occur. This broad distribution is very different from the editing sites of Sendai virus or SV5, where predominantly one form of edited mRNA containing either a one or two G insertion respectively is created, to access the single overlapping ORF of these viruses. A model is proposed to explain how paramyxoviruses control the range of G insertions on that fraction of the mRNAs where insertions occur. The bPIV3 P gene is unique as far as we know, in that a sizeable portion of the gene expresses all 3 reading frames as protein. bPIV3 apparently does this from a single editing site by removing the constraints which control the number of slippage rounds which take place.

Animals↗
PubMed1990

A stuttering model for paramyxovirus P mRNA editing.

Paramyxovirus P genes are transcribed into two mRNAs which differ from each other by either one (measles and Sendai virus) or two (SV5 and mumps virus) G insertions, and which code for either the P or V proteins. The G insertions always occur within a short run of Gs, and a stuttering mechanism for the insertions has been suggested in which the viral polymerase reiteratively copies a template C residue during mRNA synthesis. Support for this mechanism was obtained by varying the reaction conditions during Sendai virus mRNA synthesis in vitro. A stuttering model is proposed which accounts for how the ratio of inserted to uninserted mRNAs is controlled, and why some paramyxoviruses insert one G and others two Gs when insertions occur.

Base Sequence↗
PubMed1990

Development and dissemination of an aspiration risk reduction diet.

Patients with oropharyngeal swallowing disorders secondary to neurologic impairments benefit from diets that minimize the risk of aspiration. An aspiration risk reduction diet was developed from our hospital's regular menu cycle. Examples of the diet's preparation, dissemination, and use in a 600-bed acute medical/surgical teaching hospital are discussed.

Deglutition Disorders↗
PubMed1990

Overview of geriatric nutrition.

Restoration and maintenance of optimal nutritional status in the long-term care setting may enhance the quality of life for geriatric patients. The elderly are at increased risk for poor nutrition due to age-related physiological and psychosocial changes, as well as the added toll of chronic disease. All of these may have a negative impact on adequate food intake. Health care providers must identify and address nutrition-related problems in a timely manner to correct nutrient imbalances and restore nutritional well-being in this population.

Aged↗
PubMed1990

Intracellular stability of nonreplicating paramyxovirus nucleocapsids.

Using Northern blot analysis, we have demonstrated the ability of infectious measles and Sendai virus particles to rescue the intracellular replication of their homologous defective interfering (DI) nucleocapsids up to 3 days and 1 day, respectively, after initial DI infection. The half-life of the paramyxovirus DI nucleocapsids was therefore judged to be similar to that of rhabdoviruses, and to significantly differ from that of orthomyxoviruses. Moreover, we conclude that the intracellular half-life of measles virus DI nucleocapsids makes possible DI replication in the human body after vaccination with a DI-contaminated attenuated live virus, even when this vaccination represents a low multiplicity of infection.

Animals↗
PubMed1990

Anaesthesia, movement and emesis.

One hundred and eighty-two women undergoing dilatation and curettage were allocated randomly to receive premedication comprising temazepam, papaveretum-hyoscine or placebo. The temazepam recipients reported significantly fewer episodes of postoperative nausea. Movement was blamed by 66% of patients who identified a cause for nausea. These patients had higher scores on a motion sickness susceptibility questionnaire and were more likely to have been treated previously for nausea or vomiting. It may be possible to identify susceptible patients before surgery.

Adult↗
PubMed1990

Editing of the Sendai virus P/C mRNA by G insertion occurs during mRNA synthesis via a virus-encoded activity.

Two forms of the Sendai virus P/C mRNA have been predicted: one an exact copy of the viral genome, and the other with a single G insertion within a run of three G's. We directly cloned the mRNA or portions of it containing the insertion site and screened the resulting colonies with oligonucleotides that could distinguish the presence of three or four G's at this position. We found that 31% of the mRNAs did in fact contain the predicted insertion, whereas the viral genomes contained no heterogeneity at this position. A smaller fraction (7%) of the mRNA contained two to eight G's inserted at this position. The insertions also took place during RNA synthesis in vitro with purified virions but were not detected when the mRNA was expressed in vivo via a vaccinia virus recombinant. When the Sendai virus- and vaccinia virus-derived P/C mRNAs were coexpressed in the same cells under conditions in which each could be distinguished, those from the Sendai genome were altered as before, but those from the vaccinia virus genome remained unaltered. The activity that alters the mRNA is therefore likely to be coded for by the virus and cannot function in trans.

Animals↗
PubMed1990

Sendai virus P gene produces multiple proteins from overlapping open reading frames.

The Sendai virus P gene contains overlapping open reading frames (ORFs), and several unusual mechanisms are used to produce multiple proteins from all three ORFs of this gene. These include the use of a non-AUG start codon, leaky ribosomal scanning, what appears to be scanning-independent ribosomal initiation and/or ribosomal jumping, and a form of mRNA editing.

DNA-Directed RNA Polymerases↗
PubMed1989

Scanning independent ribosomal initiation of the Sendai virus Y proteins in vitro and in vivo.

The Sendai virus P/C mRNA contains five ribosomal initiation sites between positions 81 and 201 from the 5' end. One of these sites initiates in the P open reading frame (ORF) (ATG/104), whereas four initiate in the C ORF (ACG/81 and ATGs/114, 183, 201), to give a nested set of C proteins (C', C, Y1, Y2). Introduction of new ATGs or physically breaking the mRNA upstream of these natural sites was used in vitro to prevent ribosomal scanning downstream. The results suggest that a minority of the ribosomes which initiate C (ATG/114) and all of those which initiate Y1 and Y2 (ATGs/183 and 201) do so by a scanning independent mechanism. When the leaky ACG/81 site is changed to a non-leaky ATG site in in vivo experiments, ribosomal initiation at Y is again not diminished, whereas that at C as well as at P becomes undetectable. Ribosomal initiation at Y appears to be scanning independent in vitro and in vivo. That at C is partly independent in vitro, but completely dependent in vivo. These results are discussed in terms of a model of internal initiation at Y.

Codon↗
PubMed1989

Pulmonary disease in progressive systemic sclerosis. A complication of gastroesophageal reflux and occult aspiration?

Thirteen patients with progressive systemic sclerosis were studied to evaluate the possible role of gastroesophageal reflux as a contributing pathogenic factor in the pulmonary disease of the patients. The evaluation of all patients included fiberoptic esophagogastroduodenoscopy with biopsies of the esophagus, otolaryngologic evaluation, technetium Tc 99m sulfur colloid aspiration scan, pulmonary function testing, including the diffusing capacity for carbon monoxide (DLCO) test, and 24-hour intraesophageal pH monitoring with probes placed 5 and 15 cm above the lower esophageal sphincter. Eleven patients had microscopic and macroscopic evidence of proximal esophagitis, 12 patients had laryngeal changes suggestive of aspiration, and 12 patients had abnormal DLCO values. Using multiple regression analysis, the degree of DLCO impairment correlated with the proximal and distal reflux episodes and scores recorded by pH monitoring. There was direct and indirect evidence for proximal gastroesophageal reflux and aspiration in the majority of patients, and a distinct correlation between the severity of reflux and the severity of pulmonary disease. Aggressive antireflux therapy may be helpful in reducing the pulmonary damage due to aspiration in these patients.

Adult↗
PubMed1988

Ribosomal initiation from an ACG codon in the Sendai virus P/C mRNA.

The Sendai virus P/C mRNA expresses the P and C proteins from alternate reading frames. The C reading frame of this mRNA, however, is responsible for three proteins, C', C and Y, none of which appear to be precursors to each other in vivo. Using site-directed and deletion mutagenesis of the P/C gene cloned in SP6 and in vitro translation of the mRNAs, we show that the 5' most proximal initiation codon of the mRNA is an ACG at position 81, responsible for C' synthesis. The succeeding initiation codons, all ATGs, are responsible for the P protein (position 104), the C protein (position 114) and the Y protein(s) (either positions 183 or 201). Examination of the relative molar amounts of the C', P and C proteins found in vivo suggests that an ACG in an otherwise favorable context is almost as efficient for ribosome initiation as an ATG in a less favored context, but only 10-20% as efficient as an ATG in a more favored context. The judicious choice of increasingly more favorable initiation codons in the P/C gene allows multiple proteins to be made from a single mRNA.

Animals↗
PubMed1988

Scanning independent ribosomal initiation of the Sendai virus X protein.

Both peptide antisera and monoclonal antibodies detect a 10-kd protein (X) in Sendai virus infected cells, which represents approximately the last 95 residues of the 568-amino-acid-long P protein. The X protein does not appear to be derived from a precursor P in vivo, and in in vitro X can be made under conditions in which P synthesis has been arrested by hybridized DNA fragments or specific cleavage of the mRNA. X initiation is nevertheless cap dependent. Ribosomes which initiate X appear to pass directly from the cap to the initiation codon.

Animals↗