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1.  A Systematic Review of CPAP Adherence Across Age Groups: Clinical and Empiric Insights for Developing CPAP Adherence Interventions 
Sleep medicine reviews  2011;15(6):343-356.
Continuous positive airway pressure (CPAP) is a highly efficacious treatment for obstructive sleep apnea (OSA) but adherence to the treatment limits its overall effectiveness across all age groups of patients. Factors that influence adherence to CPAP include disease and patient characteristics, treatment titration procedures, technological device factors and side effects, and psychological and social factors. These influential factors have guided the development of interventions to promote CPAP adherence. Various intervention strategies have been described and include educational, technological, psychosocial, pharmacological, and multi-dimensional approaches. Though evidence to date has led to innovative strategies that address adherence in CPAP-treated children, adults, and older adults, significant opportunities exist to develop and test interventions that are clinically applicable, specific to subgroups of patients likely to demonstrate poor adherence, and address the multifactorial nature of CPAP adherence. The translation of CPAP adherence promotion interventions to clinical practice is imperative to improve health and functional outcomes in all persons with CPAP-treated OSA.
PMCID: PMC3202028  PMID: 21652236
Obstructive sleep apnea; Continuous positive airway pressure; patient compliance
2.  Randomised controlled comparison of continuous positive airways pressure, bilevel non-invasive ventilation, and standard treatment in emergency department patients with acute cardiogenic pulmonary oedema 
Emergency Medicine Journal : EMJ  2004;21(2):155-161.
Background: Continuous positive airways pressure (CPAP) and bilevel non-invasive ventilation may have beneficial effects in the treatment of patients with acute cardiogenic pulmonary oedema. The efficacy of both treatments was assessed in the UK emergency department setting, in a randomised comparison with standard oxygen therapy.
Methods: Sixty patients presenting with acidotic (pH<7.35) acute, cardiogenic pulmonary oedema, were randomly assigned conventional oxygen therapy, CPAP (10 cm H2O), or bilevel ventilation (IPAP 15 cm H2O, EPAP 5 cm H2O) provided by a standard ventilator through a face mask. The main end points were treatment success at two hours and in-hospital mortality. Analyses were by intention to treat.
Results: Treatment success (defined as all of respiratory rate<23 bpm, oxygen saturation of>90%, and arterial blood pH>7.35 (that is, reversal of acidosis), at the end of the two hour study period) occurred in three (15%) patients in the control group, seven (35%) in the CPAP group, and nine (45%) in the bilevel group (p = 0.116). Fourteen (70%) of the control group patients survived to hospital discharge, compared with 20 (100%) in the CPAP group and 15 (75%) in the bilevel group (p = 0.029; Fisher's test).
Conclusions: In this study, patients presenting with acute cardiogenic pulmonary oedema and acidosis, were more likely to survive to hospital discharge if treated with CPAP, rather than with bilevel ventilation or with conventional oxygen therapy. There was no relation between in hospital survival and early physiological changes. Survival rates were similar to other studies despite a low rate of endotracheal intubation.
PMCID: PMC1726258  PMID: 14988338
3.  Rapid screening for depression – validation of the Brief Case-Find for Depression (BCD) in medical oncology and palliative care patients 
British Journal of Cancer  2004;91(5):900-906.
PMCID: PMC2409870  PMID: 15305199
depression; mass screening; medical oncology; palliative care
4.  Management of primary and recurrent inguinal hernia by surgeons from the South West of England. 
BACKGROUND: The National Institute of Clinical Excellence (NICE) has advocated open mesh repair for primary hernia but suggested laparoscopic repair may be considered for recurrent hernias. AIM: To establish current surgical practice by surgeons from the South West of England. METHODS: A postal survey was distributed to 121 consultant surgeons and a response rate of 75% was achieved. RESULTS: The majority (86%) of the surgeons surveyed performed hernia repairs, and most (95%) of these used open mesh repair as standard for primary inguinal hernia. Only 8% used laparoscopic repair routinely for primary hernias. Few consultants (only 28%) were able to quote formally audited hernia recurrence rates. A total of 90% of respondents still employed open mesh repair routinely for recurrent hernias; however, if mesh had been used for the primary repair, this figure fell to 55%. Some 7% of respondents recommended laparoscopic repair for recurrent hernia, but this increased to 17% if the primary repair was done with mesh. All laparoscopic surgeons in the South West employed the totally extraperitoneal approach (TEP). There was a range of opinion on the technical demands of repair of a recurrent hernia previously mended with mesh; the commonest cause of mesh failure was thought to be a medial direct recurrence (insufficient mesh medially). CONCLUSIONS: Current surgical practice for primary hernias in the South West England reflects NICE guidelines although many surgeons continue to manage recurrent hernias by further open repair. In this survey, there was anecdotal evidence to suggest that hernia recurrence can be managed effectively by open repair.
PMCID: PMC1964431  PMID: 14629882
5.  Effects of Point Mutations in the Major Capsid Protein of Beet Western Yellows Virus on Capsid Formation, Virus Accumulation, and Aphid Transmission 
Journal of Virology  2003;77(5):3247-3256.
Point mutations were introduced into the major capsid protein (P3) of cloned infectious cDNA of the polerovirus beet western yellows virus (BWYV) by manipulation of cloned infectious cDNA. Seven mutations targeted sites on the S domain predicted to lie on the capsid surface. An eighth mutation eliminated two arginine residues in the R domain, which is thought to extend into the capsid interior. The effects of the mutations on virus capsid formation, virus accumulation in protoplasts and plants, and aphid transmission were tested. All of the mutants replicated in protoplasts. The S-domain mutant W166R failed to protect viral RNA from RNase attack, suggesting that this particular mutation interfered with stable capsid formation. The R-domain mutant R7A/R8A protected ∼90% of the viral RNA strand from RNase, suggesting that lower positive-charge density in the mutant capsid interior interfered with stable packaging of the complete strand into virions. Neither of these mutants systemically infected plants. The six remaining mutants properly packaged viral RNA and could invade Nicotiana clevelandii systemically following agroinfection. Mutant Q121E/N122D was poorly transmitted by aphids, implicating one or both targeted residues in virus-vector interactions. Successful transmission of mutant D172N was accompanied either by reversion to the wild type or by appearance of a second-site mutation, N137D. This finding indicates that D172 is also important for transmission but that the D172N transmission defect can be compensated for by a “reverse” substitution at another site. The results have been used to evaluate possible structural models for the BWYV capsid.
PMCID: PMC149785  PMID: 12584348
6.  P0 of Beet Western Yellows Virus Is a Suppressor of Posttranscriptional Gene Silencing 
Journal of Virology  2002;76(13):6815-6824.
Higher plants employ a homology-dependent RNA-degradation system known as posttranscriptional gene silencing (PTGS) as a defense against virus infection. Several plant viruses are known to encode proteins that can suppress PTGS. Here we show that P0 of beet western yellows virus (BWYV) displays strong silencing suppressor activity in a transient expression assay based upon its ability to inhibit PTGS of green fluorescent protein (GFP) when expressed in agro-infiltrated leaves of Nicotiana benthamiana containing a GFP transgene. PTGS suppressor activity was also observed for the P0s of two other poleroviruses, cucurbit aphid-borne yellows virus and potato leafroll virus. P0 is encoded by the 5′-proximal gene in BWYV RNA but does not accumulate to detectable levels when expressed from the genome-length RNA during infection. The low accumulation of P0 and the resulting low PTGS suppressor activity are in part a consequence of the suboptimal translation initiation context of the P0 start codon in viral RNA, although other factors, probably related to the viral replication process, also play a role. A mutation to optimize the P0 translation initiation efficiency in BWYV RNA was not stable during virus multiplication in planta. Instead, the P0 initiation codon in the progeny was frequently replaced by a less efficient initiation codon such as ACG, GTG, or ATA, indicating that there is selection against overexpression of P0 from the viral genome.
PMCID: PMC136274  PMID: 12050394
7.  Effects of Point Mutations in the Readthrough Domain of the Beet Western Yellows Virus Minor Capsid Protein on Virus Accumulation In Planta and on Transmission by Aphids 
Journal of Virology  2000;74(3):1140-1148.
Point mutations were introduced into or near five conserved sequence motifs of the readthrough domain of the beet western yellows virus minor capsid protein P74. The mutant virus was tested for its ability to accumulate efficiently in agroinfected plants and to be transmitted by its aphid vector, Myzus persicae. The stability of the mutants in the agroinfected and aphid-infected plants was followed by sequence analysis of the progeny virus. Only the mutation Y201D was found to strongly inhibit virus accumulation in planta following agroinfection, but high accumulation levels were restored by reversion or pseudoreversion at this site. Four of the five mutants were poorly aphid transmissible, but in three cases successful transmission was restored by pseudoreversion or second-site mutations. The same second-site mutations in the nonconserved motif PVT(32-34) were shown to compensate for two distinct primary mutations (R24A and E59A/D60A), one on each side of the PVT sequence. In the latter case, a second-site mutation in the PVT motif restored the ability of the virus to move from the hemocoel through the accessory salivary gland following microinjection of mutant virus into the aphid hemocoel but did not permit virus movement across the epithelium separating the intestine from the hemocoel. Successful movement of the mutant virus across both barriers was accompanied by conversion of A59 to E or T, indicating that distinct features of the readthrough domain in this region operate at different stages of the transmission process.
PMCID: PMC111448  PMID: 10627524
8.  Conformation of the 3'-end of beet necrotic yellow vein benevirus RNA 3 analysed by chemical and enzymatic probing and mutagenesis. 
Nucleic Acids Research  1997;25(23):4723-4729.
Secondary structure-sensitive chemical and enzymatic probes have been used to produce a model for the folding of the last 68 residues of the 3'-non-coding region of beet necrotic yellow vein benevirus RNA 3. The structure consists of two stem-loops separated by a single-stranded region. RNA 3-derived transcripts were produced containing mutations which either disrupted base pairing in the helices or maintained the helices but with alterations in the base pairing scheme. Other mutants contained substitutions in single-stranded regions (loops or bulged sequences). With a few exceptions all three types of mutation abolished RNA 3 replication in vivo, suggesting that both secondary structure and specific sequences are required for efficient recognition of the 3'-terminal region of RNA 3 by viral RNA-dependent RNA polymerase.
PMCID: PMC147105  PMID: 9365250
9.  The N-terminal region of the luteovirus readthrough domain determines virus binding to Buchnera GroEL and is essential for virus persistence in the aphid. 
Journal of Virology  1997;71(10):7258-7265.
Luteoviruses and the luteovirus-like pea enation mosaic virus (PEMV; genus Enamovirus) are transmitted by aphids in a circulative, nonreplicative manner. Acquired virus particles persist for several weeks in the aphid hemolymph, in which a GroEL homolog, produced by the primary endosymbiont of the aphid, is abundantly present. Six subgroup II luteoviruses and PEMV displayed a specific but differential affinity for Escherichia coli GroEL and GroEL homologs isolated from the endosymbiotic bacteria of both vector and nonvector aphid species. These observations suggest that the basic virus-binding capacity resides in a conserved region of the GroEL molecule, although other GroEL domains may influence the efficiency of binding. Purified luteovirus and enamovirus particles contain a major 22-kDa coat protein (CP) and lesser amounts of an approximately 54-kDa readthrough protein, expressed by translational readthrough of the CP into the adjacent open reading frame. Beet western yellows luteovirus (BWYV) mutants devoid of the readthrough domain (RTD) did not bind to Buchnera GroEL, demonstrating that the RTD (and not the highly conserved CP) contains the determinants for GroEL binding. In vivo studies showed that virions of these BWYV mutants were significantly less persistent in the aphid hemolymph than were virions containing the readthrough protein. These data suggest that the Buchnera GroEL-RTD interaction protects the virus from rapid degradation in the aphid. Sequence comparison analysis of the RTDs of different luteoviruses and PEMV identified conserved residues potentially important in the interaction with Buchnera GroEL.
PMCID: PMC192067  PMID: 9311800
10.  Dietary sodium intake and the risk of airway hyperreactivity in a random adult population. 
Thorax  1994;49(9):875-880.
BACKGROUND--High dietary sodium intake has been identified as a potential cause of asthma and airway hyperreactivity. This study was designed to test the hypothesis that dietary sodium intake is an independent determinant of the risk of hyperreactivity in the general population, and to assess the role of atopy in the association between these factors. METHODS--Airway reactivity to methacholine, atopy, 24 hour urinary sodium excretion, and self-reported smoking and symptom history were measured in a random sample of 1702 adults aged 18-70 from an administrative district of Nottingham. Hyperreactivity was defined as a PD20FEV1 of 12.25 mumol or less, and atopy was defined quantitatively as the mean allergen skin weal response to Dermatophagoides pteronyssinus, cat fur, and grass pollen, and categorically as the occurrence of any allergen response 1 mm or greater than the saline control. Multiple logistic regression analysis was used to estimate the independent relative odds of hyperreactivity, atopy, or symptoms in relation to sodium excretion in all 1702 subjects, and multiple linear regression to assess the independent relation between sodium excretion and mean allergen skin weal diameter, and the PD20 value amongst hyperreactive subjects. RESULTS--There was no relation between the relative odds of hyperreactivity to methacholine and 24 hour urinary sodium excretion, either before or after adjustment for age, smoking, allergen skin weal diameter, and sex, and similarly no relation if the analysis was restricted to men or women only. The relative odds of having at least one allergen skin test response 1 mm greater than the saline control were increased in relation to sodium excretion after adjustment for age, sex, and smoking by a ratio of 2.08 (95% CI 1.04 to 4.15) per log10 unit increase in sodium excretion, but there was no evidence of an association between sodium excretion and the occurrence of self-reported wheeze, hay fever, eczema, or asthma. There was no relation between 24 hour sodium excretion and the magnitude of the mean allergen skin weal response or the PD20 value. CONCLUSIONS--These findings do not support the hypothesis that a high dietary sodium intake is a risk factor for airway hyperreactivity or atopic disease in the general adult population.
PMCID: PMC475179  PMID: 7940426
11.  A cross sectional study of the independent effect of occupation on lung function in British coal miners. 
BACKGROUND: Chronic bronchitis and emphysema are now recognised complications of occupational exposure to coal dust, and since 1992 compensation has been available for miners with impaired lung function provided that they also have x ray film evidence of pneumoconiosis. However, many miners with heavy exposure to coal dust and impairment of lung function therefore do not qualify for compensation because they do not have simple pneumoconiosis. In the present study attempts were made to determine whether coal mining is an independent risk factor for impairment of lung function in a group of Nottinghamshire miners with no evidence of simple pneumoconiosis, by comparing these men with a group of local controls who were not occupationally exposed. METHOD: Forced expiratory volume in one second (FEV1) and forced vital capacity (FVC) were obtained on 1286 miners with no evidence of pneumoconiosis on x ray film. Lung function data were also obtained from a random sample of 567 men aged between 40 and 70 living in a district of Nottingham and who had never worked in the mining industry or in any other dusty occupation. Multiple linear regression in SPSS was used to estimate the mean independent effect of mining on FEV1 and FVC after adjustment for age, height, and smoking, in all miners and controls, and in a subgroup of men of 45 and under. In men of 45 and under, the independent effects of mining and smoking on the probability of a deficit of one litre or more from modelled predicted FEV1 values were computed with logistic regression in EGRET. RESULTS: There was a significant mean effect of mining on FEV1 after adjustment for age, height, and smoking of -155 ml (95% confidence interval (95% CI) -74 to -236 ml, P < 0.001), but the size of effect was inversely related to age such that in men of 45 and under the estimated mean effect of mining was -251 ml (95% CI -140 to -361 ml, P < 0.001). In this subgroup of younger men, 4.7% of miners and 0.7% of controls had a deficit of one litre or more from predicted FEV1 values, and in logistic regression, there was a marginally significant independent effect of both smoking (P = 0.05) and mining (P = 0.07) for a deficit of this magnitude. CONCLUSIONS: Occupational exposure to coal dust is associated with a small mean deficit in lung function even in the absence of simple pneumoconiosis, and independently from the effects of smoking. The requirement that miners should have evidence of pneumoconiosis to qualify for compensation for impaired lung function is therefore unjustified.
PMCID: PMC1128425  PMID: 8777449
12.  Production of extracellular slime by coryneforms colonizing hydrocephalus shunts. 
Journal of Clinical Microbiology  1994;32(7):1705-1709.
Corynebacterium spp. are responsible for an important minority of cases of colonization of cerebrospinal fluid shunts used for the treatment of hydrocephalus. In common with coagulase-negative staphylococci, they present a serious therapeutic problem because they are often resistant to multiple antibiotics. We studied the morphologies of coryneforms in colonized hydrocephalus shunts removed from patients and observed extracellular slime similar in appearance to that seen in coagulase-negative staphylococci. We also studied a series of such isolates from other cases of hydrocephalus shunt colonization using an established laboratory model and consistently observed slime production in these shunts as well. We propose that this might be a further reason for failure to eradicate these organisms without shunt removal as well as a factor in their pathogenesis in device-related infections.
PMCID: PMC263768  PMID: 7929761
13.  Perioperative Prophylactic Chemotherapy of Echinococcus Granulosus: Determination of Minimum Effective Length of Albendazole Therapy in in Vitro Protoscolex Culture 
HPB Surgery  1990;2(3):159-164.
Protoscoleces of Echinococcus granulosus were cultured in vitro in 500, 250 or 100 μg/1 albendazole sulphoxide for 1, 3, 7, 10, 14d and then ‘recued’ (R) into drug-free medium for the remainder ofthe culture period. Successful minimum lengths of therapy were much longer than for praziquantel, and only at 500μg/1 was the 10dR treatment as effective as continuous therapy for 28d. Treatment with 100 μg/1 both in continuous culture and in the ‘R’ experiments was ineffective over a 35d period. The results are compared with those from similar experiments using praziquantel.
PMCID: PMC2423571  PMID: 2278912
14.  Effect of Albendazole on Human Hydatid Cysts: An Ultrastructural Study  
HPB Surgery  1990;2(2):105-113.
Five patients with hepatic (3), pelvic (1) or spinal (1) hydatid cysts received 10 mg/kg/d albendazole for 1–3 months prior to surgery. Daughter cysts were present in the spinal hydatid and in one patient with hepatic disease. Electron microscope examination of the cyst tissue of the pelvic and the 2 hepatic cysts lacking daughter cysts showed no evidence of germinal layer, and the protoscoleces were dead. The primary cyst of the hepatic hydatid with daughter cysts (1 month therapy) was also judged dead but some pieces of the daughter cyst germinal layer appeared normal and had unaffected protoscoleces. The daughter cyst tissue of the spinal hydatid (3 month therapy) appeared normal and the protoscoleces viable. In view of the undetermined viability of human hydatids before chemotherapy, treatment of longer than 1 month is advocated for hepatic cysts, particularly if daughter cysts are present, and longer therapy is indicated for spinal disease.
PMCID: PMC2423562  PMID: 2278904
15.  Comparison of albendazole, mebendazole and praziquantel chemotherapy of Echinococcus multilocularis in a gerbil model. 
Gut  1989;30(10):1401-1405.
The efficacy of albendazole (50 mg/kg/d), mebendazole (50 mg/kg/d) and praziquantel (500 mg/kg/d) against established intraperitoneal infections of Echinococcus multilocularis in gerbils was compared by monitoring parasite weight and making ultrastructural observations on treated and untreated material. Praziquantel was the most active protoscolicidal agent, reducing protoscolex viability to less than 2%, although it did not inhibit cyst growth. Albendazole was the most effective agent in reducing cyst growth and was, when compared with other regimes significantly more effective than mebendazole (p less than 0.05), praziquantel (p less than 0.01) or untreated controls (p less than 0.01).
PMCID: PMC1434421  PMID: 2583567
16.  Immunoglobulin E anti-Staphylococcus aureus antibodies in atopic patients. 
Journal of Clinical Microbiology  1981;13(6):1046-1048.
Sera from 56 patient and normal adults were examined to quantitate total immunoglobulin E (IgE) and IgE antibodies to Staphylococcus aureus and Staphylococcus epidermidis. Patients were divided into six groups based on clinical symptoms; a seventh group consisted of normal adults. Anti-S, aureus IgE binding was significantly higher in three groups of patients (those with eczema, those with or without series staphylococcal abscesses, and allergic patients with staphylococcal skin infections) than it was in the control group. Patients with high IgE due to allergies or parasitic infections without staphylococcal infections and patients with low or normal IgE and serious staphylococcal infection showed low levels of binding. The assay measured specific binding of IgE to bacterial antigens.
PMCID: PMC273947  PMID: 7251824
18.  Nucleotide sequence and genome organization of carnation mottle virus RNA. 
Nucleic Acids Research  1985;13(18):6663-6677.
The complete nucleotide sequence of carnation mottle genomic RNA (4003 nucleotides) is presented. The sequence was determined for cloned cDNA copies of viral RNA containing over 99% of the sequence and was completed by direct sequence analysis of RNA and cDNA transcripts. The sequence contains two long open reading frames which together can account for observed translation products. One translation product would arise by suppression of an amber termination codon and the sequence raises the possibility that a second suppression event could also occur. Sequence homology exists between a portion of the carnation mottle virus sequence and that of putative RNA polymerases from other RNA viruses.
PMCID: PMC321984  PMID: 3840587
19.  Nucleotide sequence of beet western yellows virus RNA. 
Nucleic Acids Research  1988;16(21):9917-9932.
The nucleotide sequence of the genomic RNA (5641 nt) of beet western yellow virus (BWYV) isolated from lettuce has been determined and its genetic organization deduced. The sequence of the 3'terminal 2208 nt of RNA of a second BWYV isolate, obtained from sugarbeet, was also determined and was found to be very similar but not identical to that of the lettuce isolate. The complete sequence of BWYV RNA contains six long open reading frames (ORFs). A cluster of three of these ORFs, including the coat protein cistron, display extensive amino acid sequence homology with corresponding ORFs of a second luteovirus, the PAV isolate of barley yellow dwarf virus (BYDV) (1,2). The ORF corresponding to the putative viral RNA-dependant RNA polymerase, on the other hand, resembles that of southern bean mosaic virus. There is circumstantial evidence that expression of the BWYV RNA polymerase ORF may involve a translational frameshift mechanism. The ORF immediately following the coat protein cistron may be translated by in-frame readthrough of the coat protein cistron amber termination codon. Similar mechanisms have been proposed for expression of the corresponding ORFs of BYDV(PAV) (1).
PMCID: PMC338827  PMID: 3194229
20.  The secondary structure of the 5'-noncoding region of beet necrotic yellow vein virus RNA 3: evidence for a role in viral RNA replication. 
Nucleic Acids Research  1993;21(6):1389-1395.
Secondary structure-sensitive chemical and enzymatic probes have been used to produce a model for the folding of the first 312 residues of the long 5'-noncoding region of beet necrotic yellow vein virus RNA 3. The structure consists of two major domains, one of which includes long distance base-pairing interactions between two short sequence elements (Box I and Box II) situated between positions 237 and 292 and complementary elements (Box I' and II') near the 5'-terminus. Previous studies have shown that base pairing between these sequence elements (in either the plus-strand or minus-strand RNA) is important for RNA 3 accumulation during infection. RNA 3 transcripts were produced containing mutations which preferentially disrupted Box II-II' base pairing in either the plus- or minus-strand. In infection experiments, transcripts with mutations which disrupted the Box II-II' interaction in the plus-strand structure replicated less efficiently than mutants in which the Box II-II' interaction was disrupted in the minus-strand. These findings indicate that the complex 5'-proximal plus-strand structure to which the Box II-II' interaction contributes comprises at least part of the promoter for plus-strand RNA synthesis.
PMCID: PMC309323  PMID: 8464729
21.  Physical and functional heterogeneity in TYMV RNA: evidence for the existence of an independent messenger coding for coat protein. 
Nucleic Acids Research  1976;3(11):3043-3061.
Turnip yellow mosaic virus RNA can be separated into two distinct components of 2 times 10(6) and 300 000 daltons molecular weight after moderate heat treatment in the presence of SDS or EDTA. The two species cannot have arisen by accidental in vitro degradation of a larger RNA, as they both possess capped 5' ends. Analysis of the newly synthesized proteins resulting from translation of each RNA by a wheat germ extract shows that the 300 000 molecular weight RNA can be translated very efficiently into coat protein. When translated in vitro the longer RNA gave a series of high molecular weight polypeptides but only very small amounts of a polypeptide having about the same mass as the coat protein. Thus our results suggest that the small RNA is the functional messenger for coat protein synthesis in infected cells.
PMCID: PMC343150  PMID: 1005112
22.  Sequence of 1000 nucleotides at the 3' end of tobacco mosaic virus RNA. 
Nucleic Acids Research  1979;6(4):1287-1308.
The sequence of 1000 nucleotides at the 3' end of tobacco mosaic virus RNA has been determined. The sequence contains the entire coat protein cistron as well as regions to its left and right. Sequence characterization was by conventional methods for use with uniformly 32P labeled RNA complemented by newer methods for in vitro 5' and 3' 32P end-labeling of RNA and its subsequent rapid analysis. The noncoding region separating the coat protein cistron from the 3' terminus is 204 residues long and may be folded into a clover-leaf-type secondary structure. The distribution of termination codons to the left of the coat protein cistron suggests that the end of the adjacent cistron is separated from the beginning of the coat protein cistron by only two nucleotides. The subgenomic viral coat protein mRNA was isolated from infected tissue and shown to be capped. The nontranslated sequence separating the cap from the AUG initiation codon is 9 residues long and thus overlaps a portion of the adjacent cistron on the genome RNA.
PMCID: PMC327773  PMID: 109810

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