RESUMO
The replication region derived from the Corynebacterium diphtheriae-Escherichia coli plasmid pNG2 was sequenced. This 1.85-kb segment contains a single origin of DNA replication, one major open reading frame and shares no sequence homology with previously described plasmids.
Assuntos
Escherichia coli/genética , Mycobacterium/genética , Replicon , Sequência de Aminoácidos , Sequência de Bases , Genes Bacterianos , Vetores Genéticos , Dados de Sequência Molecular , Plasmídeos , Homologia de Sequência do Ácido NucleicoRESUMO
A monoclonal antibody-based sandwich enzyme immunoassay (EIA) for bovine interferon-gamma (IFN-gamma) has been developed and can be used in conjunction with a whole blood culture system to diagnose tuberculosis in cattle. During its development, normal bovine plasma samples were tested to establish background levels of circulatory IFN-gamma. Of 191 samples tested, 81 (42.4%) were positive (OD > 0.1) when tested undiluted in intact monoclonal antibody (IgG1)-coated wells compared to only 8 (4.2%) in F(ab')2-coated wells, which suggested non-specific interference in the EIA rather than circulatory IFN-gamma. Reactivity of all remaining samples was removed by diluting plasmas 1/2 with 1% casein-PBS-0.05% Tween 20 supplemented with an optimum amount (5%) of normal mouse serum (NMS). Serum pools derived from BALB/c, DBA/2, C3H/HeJ, CBA/CaH and Swiss, but not C57BL/6J, mice were found to inhibit equally the reactions of five strong false-positive bovine plasma samples but had no effect on the titre of IFN-gamma in the sample. Sera from other species tested were less effective. This suggests that the interfering factors possess a high degree of specificity, since the immunoglobulin heavy chain of IgG1 produced by all these five strains of mice are allotypically identical and different to IgG1 produced by C57BL/6J mice. The use of F(ab')2 antibody fragments to coat plate wells and sample diluent containing 5% NMS has resulted in an EIA for bovine IFN-gamma that is virtually free from false-positive reactions, has a high degree of reproducibility and a sample detection limit equivalent to approximately 80 pg/ml recombinant bovine IFN-gamma.
Assuntos
Interferon gama/sangue , Animais , Bovinos , Técnicas Imunoenzimáticas , Fragmentos Fab das Imunoglobulinas/imunologia , Especificidade da EspécieRESUMO
We report characterisation of three copies of a novel repeat sequence isolated from a Mycobacterium bovis genomic library. The repeat occurs within open reading frames, potentially encoding a conserved tandem array of a pentapeptide sequence with the consensus X-Gly-Asn-X-Gly. The tandem array is present up to five times in M. bovis and it is proposed that they may occur in a family of genes expressing functionally related proteins. We postulate that these proteins may play a role in binding of M. bovis to host cell receptors.
Assuntos
DNA Bacteriano/genética , Mycobacterium bovis/genética , Sequências Repetitivas de Ácido Nucleico , Sequência de Aminoácidos , Sequência de Bases , Biblioteca Genômica , Dados de Sequência Molecular , Fases de Leitura Aberta , Alinhamento de SequênciaRESUMO
We report characterisation of a novel repeat sequence from a Mycobacterium bovis genomic library. The highly repeated sequence belongs to a family consisting of a 24 base pair (bp) direct repeat (DR), that appears to be organized into clusters on the chromosome. We classify the 24-bp DR into the group of prokaryotic DNA repeats known as the interspersed repetitive sequence elements. The 24-bp DR will be of potential use as a DNA fingerprinting tool in epidemiological studies of M. bovis.
Assuntos
DNA Bacteriano/genética , Mycobacterium bovis/genética , Sequências Repetitivas de Ácido Nucleico , Sequência de Bases , Sequência Consenso , Biblioteca Gênica , Dados de Sequência Molecular , Mapeamento por RestriçãoRESUMO
Growth factors are theoretically promising agents for ALS therapy, but have been disappointing in subcutaneous delivery due to either toxicity or lack of major efficacy. Leukaemia inhibitory factor (LIF), was named after its effect on haemopoietic cells, and belongs to a group of cytokines which includes CNTF, IL-6, CT-1, OM and IL-11. All group members use the gp130 signal transducing subunit for intracellular signalling, but show differences in biological effect. In vitro and in vivo studies on axotomy and nerve crush models demonstrate a powerful effect of LIF in the survival of both motor and sensory neurones, while reducing denervation induced muscle atrophy. Its effects in muscle also include stimulating myoblast proliferation in vitro, and up-regulation after muscle injury. LIF will also stimulate muscle regeneration in vivo when applied exogenously after injury. In published studies of both axotomy induced neuronal death and in the Wobbler mouse models LIF is active at doses of 10 microg/kg delivered systemically, well below the expected maximum tolerated dose suggested by primate safety studies. LIF is expressed in low levels by spinal cord neurones with significant up-regulation when the neurones are damaged by BOAA toxin, an excitatory amino acid associated with a form of ALS. This augments other evidence suggesting LIF is a trauma factor playing a role in the injury response of adult neuronal tissue, and may be more effective than related growth factors. Taken together, the data suggests LIF is a physiologically relevant trophic factor with implications in clinical medicine as a therapy for ALS, and a human recombinant form (AM424), entered human clinical trials during 1998.
Assuntos
Esclerose Lateral Amiotrófica/tratamento farmacológico , Inibidores do Crescimento/uso terapêutico , Interleucina-6 , Linfocinas/uso terapêutico , Animais , Modelos Animais de Doenças , Sistemas de Liberação de Medicamentos , Inibidores do Crescimento/química , Inibidores do Crescimento/fisiologia , Humanos , Fator Inibidor de Leucemia , Linfocinas/química , Linfocinas/fisiologia , Camundongos , Camundongos Mutantes Neurológicos , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/fisiologia , Sistema Nervoso/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Receptores de Citocinas/metabolismo , Regeneração/efeitos dos fármacos , Transdução de Sinais , Superóxido Dismutase/genética , Superóxido Dismutase-1RESUMO
Conventional methods for diagnosis and epidemiological studies of bovine tuberculosis and other mycobacterial diseases are far from ideal. In the last twelve years, molecular biology has provided new approaches which have enabled detailed studies to be made of the molecular characteristics of Mycobacterium bovis, the causative organism of bovine tuberculosis. These characteristics have been investigated for their potential use in diagnosis and epidemiological studies. Restriction fragment analysis of genomic DNA from isolates of M. bovis has provided a highly discriminatory typing system which has been used extensively for epidemiological studies. DNA elements in M. bovis have been investigated for their potential use in diagnostic assays based on the polymerase chain reaction. Individual antigens of M. bovis and their reactive epitopes have been identified and investigated for their usefulness in both serological and T cell assays of animal infection. Serological assays have proven disappointing and the full potential of T cell assays has yet to be determined. Currently, T cell assays based on recombinant antigens have not been as sensitive as those based on natural antigens.
Assuntos
Antígenos de Bactérias/genética , Mycobacterium bovis/genética , Polimorfismo de Fragmento de Restrição , Tuberculose Bovina/diagnóstico , Tuberculose Bovina/epidemiologia , Animais , Bovinos , DNA Bacteriano/análise , Mycobacterium bovis/imunologia , Proteínas Recombinantes/genéticaRESUMO
When preparations containing smooth Brucella abortus lipopolysaccharide (LPS) were used as antigen in an ELISA, strong positive reactions were obtained with sera from sheep infected with Brucella melitensis or with Brucella ovis. Oxidation of the LPS with sodium metaperiodate greatly reduced the extent of the cross-reactions with antisera to B. ovis, with little effect on the reactions with antisera to smooth B. melitensis. Periodate oxidation of hot saline extract (HSX) antigen of B. ovis markedly reduced its reactivity in ELISA with anti-B. ovis sera and eliminated cross-reactivity with anti-B. melitensis sera. The reactivity of HSX was maintained after treatment with proteinase K. A simple ELISA system, in which replicate samples from a single serum dilution were tested in parallel against both B. ovis HSX antigen and periodate-oxidised smooth phase B. abortus LPS, was evaluated. It was found to discriminate well between antibodies induced by vaccination or virulent infection with B. melitensis strains and those induced by infection with B. ovis.
Assuntos
Anticorpos Antibacterianos/imunologia , Brucella/imunologia , Lipopolissacarídeos/imunologia , Ovinos/imunologia , Animais , Antígenos de Bactérias/imunologia , Brucelose/imunologia , Reações Cruzadas , Ensaio de Imunoadsorção Enzimática/veterinária , VacinaçãoRESUMO
OBJECTIVE: To test the hypothesis that, controlling for socio-demographic factors, destructive behaviour among Aboriginal and non-Aboriginal female sole parents will not be significantly different. METHOD: This study took place among an urban population of sole parents in Adelaide, South Australia, living in government housing. Two sample subsets were made up of 52 Aboriginal and 45 non-Aboriginal mothers from similar postcodes. Trained interviewers administered a questionnaire which, in addition to basic demographic data, elicited information concerning finance, housing, upbringing, experience of abuse and police interaction. The major issue of concern in the study was suicide attempt. RESULTS: 1-in-3 of the whole sample, 2-in-5 of the non-Aboriginal and 1-in-4 of the Aboriginal subset had attempted suicide at least once and half more than once. Statistical differences among 'attempters' vs. 'non-attempters', irrespective of ethnicity, included increased familial alcohol abuse, physical and sexual abuse, economic difficulty, poor self esteem and perceived discriminatory treatment by welfare agencies and, in the case of Aboriginals, by police. CONCLUSION: The social environment is critical to understanding destructive behaviour, including self-harm, regardless of culture or ethnicity. The data show that suicide attempts among female sole parents in State-housing is one of the few health indices for which Aboriginal statistics are less than for non-Aboriginals. IMPLICATIONS: It is evident that class, rather than ethnicity, better explains self-harm in this urban population. It is suggested that reluctance to access services, especially in times of crisis, relates in part to perceptions of care services and that, for Aboriginals, the value of culturally appropriate community-run services have specific public health and policy implications.
Assuntos
Havaiano Nativo ou Outro Ilhéu do Pacífico/psicologia , Pais Solteiros/psicologia , Tentativa de Suicídio/etnologia , Adulto , Comparação Transcultural , Violência Doméstica/etnologia , Violência Doméstica/estatística & dados numéricos , Feminino , Humanos , Polícia , Habitação Popular , Distribuição Aleatória , Fatores Socioeconômicos , Austrália do Sul/epidemiologia , Transtornos Relacionados ao Uso de Substâncias/etnologia , Tentativa de Suicídio/estatística & dados numéricos , Inquéritos e Questionários , População Urbana , População Branca/psicologiaRESUMO
Cytokines are major modulators of the immune system of all animals. The cloning and expression of recombinant cytokine genes have permitted the analysis of their immune function and role in the control of the immune response to disease and vaccination. While human, murine, and bovine genes have been cloned and sequenced, the cloning of ovine cytokine genes has not yet been reported. As sheep are of dominant economic importance to the Australian farming industry, it is of significance to clone and express these genes to facilitate the development of new and better vaccines and pharmaceuticals. We have initially selected ovine gamma-interferon (gamma-IFN) as a target cytokine gene. By the use of the polymerase chain reaction (PCR), using primers based on the bovine gamma-interferon sequence, we have amplified the ovine gamma-interferon gene from crude messenger RNA extracted from lymphocytes. After cloning and DNA sequencing the gene, we found that ovine gamma-IFN is 93% identical to bovine gamma-IFN in amino acid sequence. This result indicates that the PCR method will be a rapid and efficient means for cloning other ovine cytokine genes.
Assuntos
DNA/química , Interferon gama/genética , Ovinos/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , Interferon gama/química , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Homologia de Sequência do Ácido Nucleico , Ovinos/imunologiaRESUMO
OBJECTIVE: To examine the use and impact of a computer generated, patient held health record (PHR) on information sharing, responsibility sharing and preventive health care. SETTING: An academic group, private solo and private group general practice in Adelaide, South Australia. METHODS: Patients with chronic health problem(s) were randomly assigned to an experimental control or post test only group. Pre and post intervention data were collected using a standardised audit and abstraction of the patient records into a computer based record system. In addition, patient and doctor questionnaires, telephone follow ups and face to face interviews were conducted. OUTCOME MEASURES: Patient and GP use of, and satisfaction with the PHR; effectiveness of information and responsibility sharing; and uptake and performance of selected preventive health care by patient and GP. RESULTS: Seventy-two patients were recruited (29 received the PHR, and 22 each were in the control and post test only groups). The PHR was well received and used in both primary and secondary care settings. No statistically significant differences in the outcome measures were found between the groups as well as before and after the intervention (Kruskal-Wallis, p > 0.05). Data trends suggested that the PHR may increase information and responsibility sharing as well as improve patient awareness of the issues involved, with patient participation in information sharing, preventive health care and clinical decision making. Provided training and resources were made available, participating GPs believed that the computer based methodology developed was a practical option for use in practice. CONCLUSION: The computer generated PHR is an important determinant of patient participation in information and responsibility sharing, health promotion, and disease management. Implementation and evaluation studies are recommended.
Assuntos
Medicina de Família e Comunidade/organização & administração , Controle de Formulários e Registros , Sistemas Computadorizados de Registros Médicos , Participação do Paciente , Adolescente , Adulto , Idoso , Atitude do Pessoal de Saúde , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Satisfação do Paciente , Austrália do SulRESUMO
The authors outline the results of a survey, through interviews or posted questionnaires, of 122 Adelaide general practitioners. General practitioners shared or handed over most aspects of terminal care, particularly the spiritual and social aspects. Comprehensive care for these patients is directly related to the general experience of the general practitioners. The need for improved undergraduate and postgraduate education in terminal illness is stressed.
Assuntos
Medicina de Família e Comunidade , Assistência Terminal , Austrália , Serviços de Saúde Comunitária , Coleta de Dados , Humanos , Papel do Médico , Apoio SocialRESUMO
Some educational, training and politico-economic reasons for the decline in GP obstetrics in Adelaide, South Australia, are described. It is recommended that similar studies be done in other centres in Australia to evaluate the general nature of the findings.
Assuntos
Competência Clínica , Medicina de Família e Comunidade/tendências , Obstetrícia/tendências , Austrália do SulRESUMO
Three patients with superficial burns to the face and neck from benzo-quinone derivatives in the excretions of giant millipeds of the Spirobolus family are reported from the Northern District of Papua. A review is made of the clinical features and treatment of these burns.