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Biomedical subjects

R J Hayes

Publications and source records attributed to R J Hayes.

At least 73 records · Page 4Linked to original sources
PubMed1993

TGMV replication protein AL1 preferentially binds to single-stranded DNA from the common region.

The AL1 protein of tomato golden mosaic virus (TGMV) is encoded by the viral DNA and has been shown to be essential for viral DNA replication. We have over-expressed the AL1 open reading frame in E. coli and purified the protein from bacterial extracts to near homogeneity. Using various different techniques we have studied the interaction of the AL1 protein with DNA. The AL1 protein is able to bind to DNA containing the common region of the viral genome, which can be demonstrated by photochemical cross-linking. Binding is 4-fold stronger to single-stranded than to double-stranded DNA. Antibodies against the AL1 protein can be used to precipitate the protein-DNA complex. The binding to single- and double-stranded DNA is specifically to the common region since a DNA fragment unrelated to TGMV is not shifted in a gel retardation assay.

Binding Sites↗
PubMed1993

Negative sputum smear results in HIV-positive patients with pulmonary tuberculosis in Lusaka, Zambia.

During recruitment to a prospective study of tuberculosis patients in Lusaka, Zambia, 109 had pulmonary disease proven by sputum culture for Mycobacterium tuberculosis, of whom 72 were HIV-1 antibody-positive and 37 were HIV-negative. Among these culture-proven cases, 43% of the HIV-positive patients had a negative sputum smear, compared with 24% of the HIV-negative cases. There was a strong trend towards lower grade or negative sputum smear in the HIV-positive group (P = 0.003). HIV-positive cases also had lower colony counts on culture and colonies took longer to appear. The findings imply that cases of HIV-associated pulmonary tuberculosis may frequently be missed and emphasise the need for new diagnostic methods.

AIDS-Related Opportunistic Infections↗
PubMed1993

The impact of HIV on infectiousness of pulmonary tuberculosis: a community study in Zambia.

OBJECTIVE: To examine the impact of HIV on infectiousness of pulmonary tuberculosis (TB). DESIGN: A cross-sectional tuberculin survey carried out among household contacts of HIV-1-positive and negative patients with bacteriologically confirmed pulmonary TB. Contacts were also examined for active TB. SETTING: Index cases were recruited from patients attending the University Teaching Hospital in Lusaka, Zambia and household contacts were examined during visits to their homes within Lusaka. PATIENTS, PARTICIPANTS: A total of 207 contacts of 43 HIV-positive patients, and 141 contacts of 28 HIV-negative patients with pulmonary TB were examined. MAIN OUTCOME MEASURES: Proportion of contacts of HIV-positive and negative index cases with a positive tuberculin response (diameter of induration > or = 5 mm to a dose of 2 tuberculin units). RESULTS: Fifty-two per cent of contacts of HIV-positive pulmonary TB patients had a positive tuberculin response compared with 71% of contacts of HIV-negative patients (odds ratio, 0.43; 95% CI, 0.26-0.72; P < 0.001). This difference persisted after allowing for between-household variations in the tuberculin response. Tuberculin response in the contact was related to age of contact, intimacy with the index case and crowding in the household. However, the effect of HIV status of the index case was not confounded by these variables. Tuberculin response in the contact was also related to the number of bacilli seen in the sputum smear of the index case which partially explained the effect of HIV status of the index case. Active TB was diagnosed in 4% of contacts of HIV-positive and 3% of contacts of HIV-negative cases, respectively (P = 0.8). CONCLUSIONS: HIV-positive patients with pulmonary TB may be less infectious than their HIV-negative counterparts and this may partly be explained by lower bacillary load in the sputum.

Adolescent↗
PubMed1993

Determinants of condom use in 24,000 prostitute/client contacts in The Gambia.

OBJECTIVE: To determine the factors that influence condom use among prostitutes and their clients in The Gambia. DESIGN: A cohort of 181 prostitutes working in seven bars and several rural markets in The Gambia were monitored daily for 14 months. A sample of 747 clients of these prostitutes was also questioned. MAIN OUTCOME MEASURE: Proportion of sexual contacts for which a condom was used. RESULTS: Data on 24,181 sexual contacts reported by the prostitutes indicated condom use varied according to type of partner (from 84% with clients to only 4% with regular partners). Condom use with clients varied according to location (from 91% in high-class bars to 59% in rural markets), decreased from 91% with the first client of the evening to 37% with the tenth client, and from 75% with clients paying higher charges (> D19) to 52% with those paying lower charges (< D20). Condom use was not related to the socio-demographic characteristics of the prostitutes. Clients reported lower condom use than prostitutes. Clients aged 20-24 years were least likely to use condoms, while white collar workers, traders, and those paying higher charges, were more likely to use condoms. CONCLUSIONS: The level of condom use in this cohort of prostitutes was high but not consistent. Condom use was determined more by the type of establishment and the characteristics of clients, than by any fixed tendency among the prostitutes. Thus, education campaigns should be directed as much to clients as to prostitutes. In The Gambia, 'lower-class' bars and those in rural areas where prostitutes work should be a priority target.

Adolescent↗
PubMed1993

The impact of human immunodeficiency virus on presentation and diagnosis of tuberculosis in a cohort study in Zambia.

Two hundred and forty-nine patients with tuberculosis were recruited to a cohort study to investigate the interaction between tuberculosis and HIV in Lusaka, Zambia; findings at presentation are presented here. One hundred and eighty-two (73%; 95% confidence interval 67-79%) of the cases were HIV-1 antibody positive. The diagnosis of tuberculosis was confirmed by microscopy for acid-alcohol fast bacilli, culture of Mycobacterium tuberculosis, or histology in 74% of all cases. HIV negative and positive cases differed in site of disease: among HIV negative patients 72% had pulmonary disease alone, 16% extrapulmonary disease alone and 12% had both, whereas among HIV positive patients 40% had pulmonary disease alone, 34% extrapulmonary disease alone and 26% both (P < 0.001). HIV negative and positive cases were compared with regard to outcome of diagnostic procedures: 55% of HIV negative cases could be diagnosed at enrollment by sputum smear, but only 35% of HIV positive cases (P < 0.01). Among pulmonary cases confirmed by sputum culture, 76% of HIV negative patients had a positive sputum smear, compared with 57% of HIV positive patients (P = 0.09). Pleural and pericardial disease were difficult to confirm, but culture of pleural fluid was positive in 12/46 HIV positive patients, compared with 0/11 HIV negative patients. Lymph node disease was readily confirmed by biopsy. The tuberculin test was positive in only 30/110 (27%) of HIV positive cases, but in 21/38 (55%) of HIV negative cases (P < 0.01). Mycobacterium tuberculosis was cultured in 57% of HIV negative cases and 54% of HIV positive cases; no atypical mycobacteria were isolated. Initial resistance to isoniazid was present in isolates from 5% of cases with a positive culture.

Adolescent↗
PubMed1992

Cooperative binding of the red clover necrotic mosaic virus movement protein to single-stranded nucleic acids.

The movement protein of red clover necrotic mosaic dianthovirus was produced in Escherichia coli using an expression vector. Gel retardation analysis and u.v. cross-linking studies showed that the movement protein bound cooperatively to ssRNA and ssDNA, but not to dsDNA. Binding competition experiments established that the movement protein bound to ssRNA and ssDNA with similar affinities and that the binding was not sequence-specific in the experimental conditions employed. A truncated movement protein lacking the C-terminal 88 amino acids was also shown to bind to ssRNA.

Binding, Competitive↗
PubMed1992

Complete replication of a satellite RNA in vitro by a purified RNA-dependent RNA polymerase.

The 334 nucleotide R satellite RNA was used as a template for purified RNA-dependent RNA polymerase (RdRp) from cucumber mosaic virus-infected tobacco plants. The products of the reaction were dsRNA and positive-strand RNA of the same size as the R satellite RNA. Similar products were obtained when T7 RNA polymerase positive-strand transcripts of a cDNA clone of the satellite RNA, designed to have the same 5' and 3' ends as the satellite RNA, were used as templates. The formation of the positive strands demonstrates complete replication of the satellite RNA. A positive-strand transcript with 65 and 255 additional nucleotides at the 5' and 3' ends of the satellite RNA respectively was also utilized as a template by the RdRp, but only dsRNA was formed. However, no products could be detected when the RdRp was programmed with transcripts corresponding to the negative-strand satellite RNA, either with no additional terminal nucleotides or with 24 and 310 additional nucleotides at the 5' and 3' ends respectively.

Base Sequence↗
PubMed1992

Case-control studies of severe malaria.

The majority of children infected with Plasmodium falciparum in areas of stable endemicity do not develop severe, life-threatening disease. It is important to identify risk factors for the minority who do. Case-control studies in which children with severe disease are compared with children with non-severe disease and with community controls, avoid some of the ethical and logistical problems inherent in such an undertaking. This paper discusses methodological aspects of case-control studies of severe malaria including case and control definitions, selection of cases and controls, potential risk factors, sample size calculations and analysis. Although specifically concerned with malaria, many of these issues are equally relevant to case-control studies of other infectious and parasitic diseases in a tropical environment.

Case-Control Studies↗
PubMed1990

Complete replication of a eukaryotic virus RNA in vitro by a purified RNA-dependent RNA polymerase.

A soluble RNA-dependent RNA polymerase was isolated from Nicotiana tabacum plants infected with cucumber mosaic virus (CMV), which has a genome of three positive-strand RNA components, 1, 2, and 3. The purified polymerase contained two virus-encoded polypeptides and one host polypeptide. Polymerase activity was completely dependent on addition of CMV RNA as template, and the products of reaction were single-stranded (ss) RNA and double-stranded (ds) RNA, corresponding to RNAs 1, 2, and 3, and a subgenomic RNA (RNA 4) derived from RNA 3. The ratio of ssRNA to dsRNA was about 5:1, and the ssRNA was shown to be predominantly the positive strand. This demonstrates the complete replication of a eukaryotic virus RNA in vitro by a template-dependent RNA polymerase.

Base Sequence↗
PubMed1990

Impact of HIV on tuberculosis in Zambia: a cross sectional study.

OBJECTIVE: To examine the contribution of HIV infection to the apparently increasing incidence of tuberculosis in central Africa. DESIGN: Cross sectional study. SETTING: Outpatient clinic in teaching hospital, Lusaka, Zambia. PATIENTS: 346 Adult patients with tuberculosis. RESULTS: Overall, 206 patients (60%; 95% confidence interval 54% to 65%) were positive for HIV: in one or both assays used. The peaks for both tuberculosis and HIV infection were among men aged 25-34 years and women aged 14-24 years. Of patients with confirmed pulmonary tuberculosis, 73/149 (49%; 41% to 57%) were positive for HIV; 67/83 (81%; 70% to 89%) patients with pleural disease and 16/19 (84%; 60% to 97%) patients with pericardial disease were positive. HIV positive patients with positive sputum culture were less likely to have had a positive sputum smear, and their chest x ray films less often showed classic upper zone disease or cavitation. Of 72 patients who fulfilled clinical criteria for AIDS, 17 were negative for HIV. CONCLUSIONS: The high prevalence of HIV in patients with tuberculosis suggests that an epidemic of reactivating tuberculosis is arising in those who are infected with HIV. The redirection of public health priorities towards tuberculosis would focus on a major treatable and preventable complication of the AIDS epidemic.

Adolescent↗
PubMed1990

Leptospiral infection: a household serosurvey in urban and rural communities in Barbados and Trinidad.

A longitudinal study of leptospiral agglutinins in subjects five years of age and over was undertaken in Trinidad and Barbados between 1980 and 1982. Households were sampled randomly from one urban and two rural communities on each island, giving a total of 576 eligible individuals in Barbados and 524 in Trinidad. Participants were examined three times at approximately annual intervals. The prevalence of seropositivity at a titre of 1:50 using the microscopic agglutination test was 18.5% in Barbados and 21.9% in Trinidad. Prevalence increased steeply with age in both sexes and was higher in males than females on both islands. There was a marked difference in predominating serogroups on the two islands--Autumnalis (42% of positive cases) predominated in Barbados while Bataviae (29% of positive cases) predominated in Trinidad. Estimates of incidence rates for seroconversion were 2.9% per annum for Barbados and 3.5% per annum for Trinidad. Occupational risk varied between the islands, but in both cases highest seropositivity rates (greater than 50%) were found in outdoor labourers and lowest were found in indoor non-manual workers and urban homeworkers. In Barbados seroprevalence was higher among persons who cleared drains or who had contact with livestock. Lack of an inside toilet was associated with an increase in seropositivity on both islands. There was little evidence of household clustering of seropositive cases.

Adolescent↗
PubMed1990

Epidemiological methods to study the interaction between HIV infection and other sexually transmitted diseases.

Numerous studies performed over the past 5 years have indicated an association between HIV infection and other sexually transmitted diseases (STDs), particularly those involving genital ulceration. Such an association may be causal, indicating that STDs increase susceptibility to, or infectivity of, HIV infection or may result, in whole or in part, from the mutual dependence of HIV and STDs on patterns of sexual activity, or from an effect of HIV infection on the clinical course of STDs. In this paper we discuss the issues arising in the design and analysis of studies conducted to investigate this association. A numerical illustration is used to demonstrate non-causal associations that may arise in observational studies due to confounding and misclassification. Published cross-sectional and longitudinal studies are reviewed, and recommendations made for future studies. Special emphasis is given to the use of randomized intervention trials to overcome many of the biases associated with observational studies, and to provide information on the efficacy of intensive STD treatment programmes in reducing the transmission of HIV.

Cross-Sectional Studies↗
PubMed1990

Infectious cucumber mosaic virus RNA transcribed in vitro from clones obtained from cDNA amplified using the polymerase chain reaction.

Full-length cDNA to RNA 1, RNA 2 and RNA 3 of cucumber mosaic virus strain Q (CMV-Q) was amplified using the polymerase chain reaction (PCR). The first-strand primer contained a BamHI site and sequences complementary to the 3' terminus of the RNA. The second-strand primers contained a BamHI site, a T7 promoter and sequences corresponding to the 5' terminus of each RNA. After cleavage with BamHI, the PCR products were cloned into the BamHI site of the vector pEMBL9(+). Five clones of each RNA were selected and RNA transcripts were synthesized in vitro from each clone using T7 RNA polymerase. The constructs were designed to allow transcription to initiate precisely at the 5' terminus of each RNA. All the transcripts were found to be infectious when inoculated onto Nicotiana tabacum cv. Samsun plants in sets of three, corresponding to RNA 1, RNA 2 and RNA 3. Of the transcript sets, four induced symptoms indistinguishable from symptoms induced by CMV-Q RNAs. However a fifth transcript set induced much more severe symptoms. Plasmids were also constructed to allow synthesis of transcripts with one or two additional G residues at the 5' terminus of each RNA. Although the yields of such transcripts synthesized in vitro with T7 RNA polymerase were higher, their infectivity was lower than that of those with no additional residues at their 5' termini.

Base Sequence↗
PubMed1990

[Estimate of the incidence rates of chronic parasitosis and infections based on the prevalence: Chagas disease in Latin America].

A simple method is described for using prevalence data to estimate the incidence of chronic infections. The incidence rate is estimated as the prevalence rate multiplied by the crude birth rate. The method is applicable to infections which are detectable for life and which have little effect on mortality. Strictly, it is valid only if infection occurs at or shortly after birth, but we show that little error results if infections occur gradually over the first two decades of life. A method of refining the incidence estimate, when age-specific prevalence data are available, is also given. The methods are illustrated using data on Chagas' disease in Latin America.

Age Factors↗
PubMed1990

Testing for the random occurrence of sickle cell disease in a study of 100,000 Jamaican newborns.

In a screening programme for sickle cell haemoglobinopathies at a maternity hospital in Kingston, Jamaica, 100,000 newborns were screened with the detection of 315 cases of SS disease, and 201 cases of SC disease. There was no upward or downward trend in the frequency of SS or SC births during the screening programme. The sequence numbers of the SS and SC births were analysed to examine whether these cases were randomly distributed among the 100,000 newborns. The occurrence of SS births seems to accord well with the random model but there was an excess of very long intervals between SC births.

Anemia, Sickle Cell↗