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1.
J Vet Pharmacol Ther ; 36(4): 382-8, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22943152

RESUMO

This study investigated the effects of cortisol and insulin, hormones that affect both glycaemic status and vascular function, on the in vitro contractility of isolated healthy equine small laminar veins. Small veins (150-500 µm) draining the digital laminae from healthy horses or ponies were investigated by wire myography. Concentration response curves were constructed for noradrenaline (NA), phenylephrine (PE), endothelin-1 (ET-1) and 5-hydroxytryptamine (5-HT) in the presence of either cortisol (10(-6 ) m) or insulin (1000 µIU/mL). Cortisol significantly increased the maximum contractility of laminar veins to the vasoconstrictors NA and 5-HT but decreased the maximal contraction to ET-1. Insulin decreased the contractility of vessels to PE and ET-1. It is possible that short-term cortisol excess could enhance venoconstrictor responses to 5-HT and NA in laminar veins in vivo, thereby predisposing to laminitis. Additionally, a reduction in the ability of insulin to counteract alpha-adrenoreceptor and ET-1-mediated contraction, likely to occur in subjects with insulin resistance, may further exacerbate venoconstriction in animals prone to laminitis. These mechanisms may also predispose horses with disorders such as equine Cushing's disease and equine metabolic syndrome to laminitis.


Assuntos
Casco e Garras/irrigação sanguínea , Doenças dos Cavalos/etiologia , Hidrocortisona/farmacologia , Insulina/farmacologia , Vasoconstrição/efeitos dos fármacos , Veias/efeitos dos fármacos , Animais , Endotelina-1/farmacologia , Casco e Garras/patologia , Doenças dos Cavalos/metabolismo , Cavalos , Inflamação/etiologia , Inflamação/metabolismo , Inflamação/veterinária , Norepinefrina/farmacologia , Fenilefrina/farmacologia , Serotonina/farmacologia
2.
Domest Anim Endocrinol ; 76: 106623, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-33774426

RESUMO

Total thyroxine (T4) concentrations are lower in healthy greyhounds compared to most other non-sighthound breeds. In humans, variations in the structure or concentration of the major thyroid hormone binding proteins are responsible for most reported differences between total T4 concentrations in healthy individuals from different ethnic groups or other subpopulations. The aim of this study was to determine if such variations are also responsible for the lower total T4 concentrations in greyhounds. The predicted protein sequences of thyroxine-binding globulin (TBG), transthyretin and albumin were determined in liver tissue from a euthyroid greyhound with decreased T4 concentration and a Jack Russell terrier using reverse-transcriptase PCR. Sequences were compared to each other and online reference sequences. Serum proteins from 21 greyhounds and 21 non-sighthound dogs were separated by denaturing electrophoresis and immunoblots probed with polyclonal antibodies to human TBG and transthyretin. Reactive bands were quantified by densitrometry, expressed relative to the mean of reference samples included in each gel. Serum albumin concentrations were measured using a commercially-available assay. Several SNPs were identified but none was thought likely to explain the lower total T4 concentrations in greyhounds. There was no significant difference between the quantity of any of the binding proteins in serum from greyhounds and non-sighthound dogs. However, total T4 and transthyretin concentrations were highly correlated in the greyhound group (r = 0.73, P = 0.0002). Variation in the sequence of thyroid hormone binding proteins is not responsible for low greyhound total T4 concentrations. Further evaluation of the role of transthyretin is warranted.


Assuntos
Hormônios Tireóideos , Tiroxina , Animais , Anticorpos , Cães
3.
J Vis Exp ; (174)2021 08 14.
Artigo em Inglês | MEDLINE | ID: mdl-34459812

RESUMO

Leptospirosis is a global neglected zoonosis, responsible for at least 1 million cases per year and almost 60 thousand deaths. The disease is caused by pathogenic and virulent bacteria of the genus Leptospira, either by direct contact with the bacteria or indirectly by exposure to contaminated water or soil. Domestic and wild animals act as reservoir hosts of infection, shedding leptospires from colonized renal tubules of the kidney, via urine, into the environment. The generation of mutant strains of Leptospira is critical to evaluate and understand pathogenic mechanisms of infection. CRISPR interference (CRISPRi) has proven to be a straightforward, affordable, and specific tool for gene silencing in pathogenic Leptospira. Therefore, the methodological details of obtaining the plasmid constructs containing both dCas9 and guide RNA, delivery of plasmids to Leptospira by conjugation with the E. coli strain ß2163, and transconjugant recovery and evaluation, will be described. In addition, the recently described Hornsby-Alt-Nally (HAN) media allows for the relatively rapid isolation and selection of mutant colonies on agar plates.


Assuntos
Leptospira , Leptospirose , Animais , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas , Escherichia coli , Inativação Gênica , Leptospira/genética , Leptospirose/genética
4.
Sci Rep ; 11(1): 1768, 2021 01 19.
Artigo em Inglês | MEDLINE | ID: mdl-33469138

RESUMO

Leptospirosis is a neglected, widespread zoonosis caused by pathogenic species of the genus Leptospira, and is responsible for 60,000 deaths per year. Pathogenic mechanisms of leptospirosis remain poorly understood mainly because targeted mutations or gene silencing in pathogenic Leptospira continues to be inherently inefficient, laborious, costly and difficult to implement. In addition, pathogenic leptospires are highly fastidious and the selection of mutants on solid agar media can take up to 6 weeks. The catalytically inactive Cas9 (dCas9) is an RNA-guided DNA-binding protein from the Streptococcus pyogenes CRISPR/Cas system and can be used for gene silencing, in a strategy termed CRISPR interference (CRISPRi). Here, this technique was employed to silence genes encoding major outer membrane proteins of pathogenic L. interrogans. Conjugation protocols were optimized using the newly described HAN media modified for rapid mutant recovery at 37 °C in 3% CO2 within 8 days. Complete silencing of LipL32 and concomitant and complete silencing of both LigA and LigB outer membrane proteins were achieved, revealing for the first time that Lig proteins are involved in pathogenic Leptospira serum resistance. Gene silencing in pathogenic leptospires and rapid mutant recovery will facilitate novel studies to further evaluate and understand pathogenic mechanisms of leptospirosis.


Assuntos
Antígenos de Bactérias/genética , Proteínas da Membrana Bacteriana Externa/genética , Sistemas CRISPR-Cas/genética , Inativação Gênica , Leptospira interrogans/genética , Lipoproteínas/genética , Humanos , Leptospirose/microbiologia , Fenótipo , RNA Guia de Cinetoplastídeos/genética
5.
Eur J Clin Microbiol Infect Dis ; 29(10): 1305-9, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20559675

RESUMO

Leptospirosis is a global zoonotic disease. Pathogenic Leptospira species, the causative agent of leptospirosis, colonize the renal tubules of chronically infected maintenance hosts such as dogs, rats and cattle. Maintenance hosts typically remain clinically asymptomatic and shed leptospires into the environment via urine. In contrast, accidental hosts such as humans can suffer severe acute forms of the disease. Infection results from direct contact with infected urine or indirectly, through contaminated water sources. In this study, a quantitative real-time PCR specific for lipL32 was designed to detect the urinary shedding of leptospires from dogs. The sensitivity and specificity of the assay was evaluated using both a panel of pathogenic Leptospira species and clinical microbial isolates, and samples of urine collected from experimentally infected rats and non-infected controls. The lower limit of detection was approximately 3 genome equivalents per reaction. The assay was applied to canine urine samples collected from local dog sanctuaries and the University Veterinary Hospital (UVH) at University College Dublin. Of 525 canine urine samples assayed, 37 were positive, indicating a prevalence of urinary shedding of leptospires of 7.05%. These results highlight the need to provide effective canine vaccination strategies and raise public health awareness.


Assuntos
Carga Bacteriana/métodos , Doenças do Cão/diagnóstico , Doenças do Cão/microbiologia , Leptospira/isolamento & purificação , Leptospirose/veterinária , Urina/microbiologia , Animais , Proteínas da Membrana Bacteriana Externa/genética , Derrame de Bactérias , DNA Bacteriano/genética , Cães , Hospitais Veterinários , Irlanda , Leptospirose/diagnóstico , Leptospirose/microbiologia , Lipoproteínas/genética , Reação em Cadeia da Polimerase/métodos , Prevalência , Ratos , Sensibilidade e Especificidade
6.
J Clin Microbiol ; 47(4): 1199-201, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19204100

RESUMO

Spirochetes of the genus Treponema were cultured from 7 of 10 cases of digital dermatitis in sheep. Two cultures comprised Treponema phagedenis-like and Treponema medium/Treponema vincentii-like spirochetes, respectively, while the remaining cultures comprised mixed populations of Treponema medium/Treponema vincentii-like, Treponema phagedenis-like, and Treponema denticola/Treponema putidum-like organisms.


Assuntos
Dermatite/veterinária , Doenças dos Ovinos/microbiologia , Dermatopatias Bacterianas/veterinária , Treponema/classificação , Treponema/isolamento & purificação , Animais , DNA Bacteriano/química , DNA Bacteriano/genética , Dermatite/microbiologia , Dados de Sequência Molecular , Análise de Sequência de DNA , Ovinos , Dermatopatias Bacterianas/microbiologia , Treponema/genética
7.
J Appl Microbiol ; 107(3): 707-16, 2009 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-19302325

RESUMO

Summary Rats, dogs, cattle, bats and sea lions, exemplify the diversity of mammalian species that can facilitate transmission of the zoonotic disease leptospirosis. The causative agent, pathogenic species of Leptospira, is shed in urine of chronically infected hosts. Direct contact with infected urine, or indirectly with water sources contaminated with infected urine, poses a risk of infection for humans exposed during water-related recreational and occupational activities. New serovars of Leptospira and maintenance hosts continue to be identified. In the western world, incidences of recreational exposure are increasing, while incidences of occupational exposure are decreasing. Adventure travellers returning from tropical regions, are presenting at clinics with symptoms of leptospirosis following participation in high risk activities including white water rafting, triathlons, endurance races and caving. Risks of infection can be reduced with increased awareness of how the disease is contracted, by avoiding contact with high risk water sources and the use of prophylaxis during high risk activities. Molecular techniques can be used to provide risk assessments prior to competition, to supplement epidemiology, and to assess shedding of Leptospira in urine samples.


Assuntos
Exposição Ambiental , Leptospirose/transmissão , Recreação , Adulto , Animais , Animais Domésticos , Animais Selvagens , Bovinos , Cães , Feminino , Humanos , Leptospirose/prevenção & controle , Masculino , Camundongos , Pessoa de Meia-Idade , Ratos , Fatores de Risco , Rios
8.
Vet Pathol ; 46(5): 792-9, 2009 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-19429975

RESUMO

Pathogenic species of Leptospira cause leptospirosis, a global zoonotic disease. Leptospira colonize renal tubules of chronically infected maintenance hosts, from where they are shed in urine to the environment and survive in suitable moist conditions. Transmission of disease to new hosts is facilitated by contact with contaminated urine or water sources, because Leptospira can penetrate broken skin or mucosal surfaces of new hosts. Infection of new hosts may be asymptomatic, as with chronically infected maintenance hosts, or may result in an acute disease process in which clinical signs can include fever, jaundice, renal failure, and pulmonary hemorrhage. Those factors that determine if an animal will suffer an acute or a chronic infection are not fully understood but include host animal species, infecting serovar, and infecting dose. During chronic infection, renal colonization and leptospiruria persist despite cellular and humoral responses by the host. Tubulointerstitial nephritis is the most common lesion associated with chronic infection, and this may progress to fibrosis and subsequent renal failure. This review aims to address how Leptospira cause tubulointerstitial nephritis during chronic leptospirosis and to summarize the mechanisms by which Leptospira might evade host immune responses during chronic colonization of the renal tubule.


Assuntos
Interações Hospedeiro-Patógeno/imunologia , Nefropatias/veterinária , Leptospira/imunologia , Leptospirose/veterinária , Zoonoses/parasitologia , Animais , Reservatórios de Doenças/microbiologia , Reservatórios de Doenças/veterinária , Predisposição Genética para Doença , Nefropatias/imunologia , Nefropatias/microbiologia , Leptospirose/imunologia , Leptospirose/microbiologia , Leptospirose/transmissão , Receptores Toll-Like/imunologia , Zoonoses/transmissão
9.
Vaccine ; 37(36): 5428-5438, 2019 08 23.
Artigo em Inglês | MEDLINE | ID: mdl-31375438

RESUMO

MIP and CPAF from Chlamydia have been shown to be effective in inducing immune responses important in clearing chlamydial infections. This study evaluates the protection conferred by MIP and CPAF as novel vaccines in pregnant C. abortus challenged ewes. Fifty C. abortus sero-negative sheep were randomly allocated into 5 groups of 10 according to the treatment they were to receive (1) 100 µg of MBP-MIP (2) 100 µg CPAF (3) 50 µg MBP-MIP and 50 µg CPAF (4) Tris-buffer (negative control) (5) Enzovax (positive control). Booster inoculations were administered 3 weeks after primary inoculations. Blood samples were taken pre-vaccination and weekly for 5 weeks. Five months after vaccination the ewes were mated. Pregnant ewes were then challenged on day 90 of gestation. Blood samples taken at four time-points post challenge were analysed for IFNγ levels, TNFα and IL-10 expression and anti-chlamydial antibody levels. Vaginal swabs, placental and foetal tissue and bacterial shedding were analysed using qPCR to quantify levels of C. abortus. Enzovax was 100% effective with no abortions occurring. The MIP/CPAF combined vaccine offered the greatest protection of the novel vaccines with 67% of ewes giving birth to one or more live lambs equating to a 50% vaccine efficacy rate. MIP and CPAF administered singly did not confer protection. Enzovax and MIP/CPAF vaccinated ewes had longer gestations and lambs with higher birth weights than negative control ewes. Aborting ewes shed higher numbers of C. abortus than ewes that had live lambs, all vaccinated ewes demonstrated lower levels of bacterial shedding than negative control ewes with Enzovax ewes shedding significantly fewer bacteria. Ewes that went on to abort had significantly higher levels of IFNγ and IL-10 at day 35 post challenge and significantly higher levels of anti-chlamydial antibodies at 24 h post lambing compared to ewes that had live lambs.


Assuntos
Infecções por Chlamydia/imunologia , Infecções por Chlamydia/prevenção & controle , Chlamydia/imunologia , Chlamydia/patogenicidade , Endopeptidases/imunologia , Vacinação/métodos , Aborto Animal/prevenção & controle , Animais , Vacinas Bacterianas/imunologia , Vacinas Bacterianas/uso terapêutico , Endopeptidases/metabolismo , Feminino , Gravidez , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/metabolismo , Ovinos , Doenças dos Ovinos/imunologia , Doenças dos Ovinos/prevenção & controle
10.
Appl Environ Microbiol ; 74(12): 3783-94, 2008 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-18441119

RESUMO

Nucleotide polymorphism associated with the O-antigen-encoding locus, rfb, in Enterobacter sakazakii was determined by PCR-restriction fragment length polymorphism analysis. Based on the analysis of these DNA profiles, 12 unique banding patterns were detected among a collection of 62 strains from diverse origins. Two common profiles were identified and were designated serotypes O:1 and O:2. DNA sequencing of the 12,500-bp region flanked by galF and gnd identified 11 open reading frames, all with the same transcriptional direction. Analysis of the proximal region of both sequences demonstrated remarkable heterogeneity. A PCR assay targeting genes specific for the two prominent serotypes was developed and applied for the identification of these strains recovered from food, environmental, and clinical samples.


Assuntos
Cronobacter sakazakii/classificação , Cronobacter sakazakii/genética , Antígenos O/genética , Proteínas de Bactérias/genética , Análise por Conglomerados , Cronobacter sakazakii/isolamento & purificação , Impressões Digitais de DNA , DNA Bacteriano/química , DNA Bacteriano/genética , Infecções por Enterobacteriaceae/microbiologia , Microbiologia Ambiental , Microbiologia de Alimentos , Ordem dos Genes , Genótipo , Humanos , Dados de Sequência Molecular , Fases de Leitura Aberta , Reação em Cadeia da Polimerase/métodos , Polimorfismo de Fragmento de Restrição , Análise de Sequência de DNA , Transcrição Gênica
11.
Equine Vet J ; 40(5): 488-92, 2008 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-18487099

RESUMO

REASONS FOR PERFORMING STUDY: Endothelin-1 (ET-1) may be a key mediator in the pathogenesis of laminitis, but endothelin-mediated responses in the venous microcirculation of the equine foot have yet to be fully characterised. OBJECTIVES: To characterise the response of equine laminar veins to ET-1 and evaluate the ET-1 receptor subtypes that mediate this response. METHODS: Small veins (150-500 microns) draining the equine digital laminae from healthy horses and ponies subjected to euthanasia at an abattoir were investigated using wire myography. Concentration response curves were constructed for ET-1 in the presence of ETA (BQ123) and ETB (BQ788) receptor antagonists, and L-NAME, a nitric oxide synthase blocker. The selective ETB receptor agonist BQ3020 was investigated alone and following incubation with L-NAME, with or without BQ788. RESULTS: Endothelin-1 contraction of laminar veins was significantly inhibited by BQ123 but not by BQ788. In the presence of L-NAME, sensitivity of laminar veins to ET-1 was enhanced 4-fold, and further addition of BQ788 did not alter this increased sensitivity. BQ3020 induced no venoconstriction; however, in the presence of L-NAME, it caused contraction of veins with approximately 30% of the efficacy of ET-1. The action of BQ3020 in the presence of L-NAME was abolished by BQ788. CONCLUSIONS: Both ETA and ETB receptors are involved in the net tonic response to ET-1 in normal laminar veins. A population of ETB receptors may be present on the vascular endothelium and on smooth muscle of laminar veins, and the action of ET-1 at these 2 sites is likely to be approximately equal and opposite. POTENTIAL RELEVANCE: Our results clarify the function of the ET-1 receptor subtypes in laminar veins from healthy horses. Further study of ET-1 receptors in laminitic horses is therefore warranted.


Assuntos
Endotelina-1/metabolismo , Casco e Garras/irrigação sanguínea , Contração Muscular/fisiologia , Receptor de Endotelina A/fisiologia , Receptor de Endotelina B/fisiologia , Animais , Antagonistas do Receptor de Endotelina A , Antagonistas do Receptor de Endotelina B , Endotelina-1/farmacologia , Endotélio Vascular/metabolismo , Endotélio Vascular/fisiologia , Doenças do Pé/metabolismo , Doenças do Pé/veterinária , Doenças dos Cavalos/metabolismo , Cavalos , Coxeadura Animal/metabolismo , Contração Muscular/efeitos dos fármacos , Músculo Liso Vascular/efeitos dos fármacos , Músculo Liso Vascular/fisiologia , NG-Nitroarginina Metil Éster , Oligopeptídeos/farmacologia , Peptídeos Cíclicos/farmacologia , Piperidinas/farmacologia , Veias/efeitos dos fármacos , Veias/fisiologia
12.
Transbound Emerg Dis ; 63(2): e178-84, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25065690

RESUMO

Leptospirosis is a neglected zoonosis of global importance with a complex epidemiology that affects humans, domestic and wild mammals. However, due to the diversity of clinical signs and difficulties of establishing a confirmatory laboratory diagnosis, the disease remains poorly investigated, particularly in the developing world. In Morocco, a descriptive study of the seroprevalence of Leptospira infection in animals has never been undertaken. To fill this gap, the current study was conducted on a subset of animals in north-west Morocco as a preliminary step towards understanding the epidemiological patterns of animal leptospirosis in the country. The study was conducted on 289 serum samples collected between January and April 2012 from dogs, cattle, sheep, goats and donkeys in the areas of Rabat-Temara, Sidi Kacem and Oulmes. All serum samples were tested by the MAT with 14 reference strains of the most prevalent pathogenic serovars of Leptospira and two serovars of non-pathogenic Leptospira. The overall seroprevalence of Leptospira in cattle, sheep, goats, dogs and donkeys was 15%, 18%, 20%, 21% and 20%, respectively. The most prevalent serogroups found in each species were Ballum, Sejroe, and Australis in cattle, Ballum, Australis and Sejroe in sheep, Australis and Ballum in goats, Javanica and Australis in donkey and Australis, Ballum and Canicola in dogs. Of all the serogroups tested in this study, Icterohaemorrhagiae, the only serogroup which has been previously reported in humans in Morocco, was rarely reactive. The majority of reactive sera were collected from low land areas. A large number of sera samples classified as seronegative when tested against pathogenic leptospires were positive when tested against non-pathogenic leptospires; this is suggestive of possible novel, as yet unclassified, Leptospira serovars in Morocco. Eleven of thirteen sheep urine samples were positive by real-time PCR confirming their role as Leptospira carriers in Morocco.


Assuntos
Animais Domésticos/microbiologia , Leptospira/imunologia , Leptospirose/epidemiologia , Animais , Anticorpos Antibacterianos/sangue , Bovinos , Cães , Equidae , Cabras , Leptospira/classificação , Leptospira/isolamento & purificação , Leptospirose/microbiologia , Leptospirose/veterinária , Marrocos/epidemiologia , Prevalência , Reação em Cadeia da Polimerase em Tempo Real , Estudos Soroepidemiológicos , Sorogrupo , Ovinos
13.
J Small Anim Pract ; 56(3): 159-79, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25754092

RESUMO

Leptospirosis is a zoonotic disease with a worldwide distribution affecting most mammalian species. Clinical leptospirosis is common in dogs but appears to be rare in cats. Both dogs and cats, however, can shed leptospires in the urine. This is problematic as it can lead to exposure of humans. The control of leptospirosis, therefore, is important not only from an animal but also from a public health perspective. The aim of this consensus statement is to raise awareness of leptospirosis and to outline the current knowledge on the epidemiology, clinical features, diagnostic tools, prevention and treatment measures relevant to canine and feline leptospirosis in Europe.


Assuntos
Doenças do Gato/prevenção & controle , Doenças do Cão/prevenção & controle , Leptospirose/veterinária , Animais , Antibacterianos/uso terapêutico , Doenças do Gato/diagnóstico , Doenças do Gato/tratamento farmacológico , Doenças do Gato/microbiologia , Doenças do Gato/transmissão , Gatos/microbiologia , Doenças do Cão/diagnóstico , Doenças do Cão/tratamento farmacológico , Doenças do Cão/microbiologia , Doenças do Cão/transmissão , Cães/microbiologia , Humanos , Leptospira , Leptospirose/diagnóstico , Leptospirose/tratamento farmacológico , Leptospirose/microbiologia , Leptospirose/prevenção & controle , Leptospirose/transmissão , Zoonoses/microbiologia , Zoonoses/prevenção & controle , Zoonoses/transmissão
14.
Animal ; 9(1): 1-17, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25359324

RESUMO

Animal production and health (APH) is an important sector in the world economy, representing a large proportion of the budget of all member states in the European Union and in other continents. APH is a highly competitive sector with a strong emphasis on innovation and, albeit with country to country variations, on scientific research. Proteomics (the study of all proteins present in a given tissue or fluid - i.e. the proteome) has an enormous potential when applied to APH. Nevertheless, for a variety of reasons and in contrast to disciplines such as plant sciences or human biomedicine, such potential is only now being tapped. To counter such limited usage, 6 years ago we created a consortium dedicated to the applications of Proteomics to APH, specifically in the form of a Cooperation in Science and Technology (COST) Action, termed FA1002--Proteomics in Farm Animals: www.cost-faproteomics.org. In 4 years, the consortium quickly enlarged to a total of 31 countries in Europe, as well as Israel, Argentina, Australia and New Zealand. This article has a triple purpose. First, we aim to provide clear examples on the applications and benefits of the use of proteomics in all aspects related to APH. Second, we provide insights and possibilities on the new trends and objectives for APH proteomics applications and technologies for the years to come. Finally, we provide an overview and balance of the major activities and accomplishments of the COST Action on Farm Animal Proteomics. These include activities such as the organization of seminars, workshops and major scientific conferences, organization of summer schools, financing Short-Term Scientific Missions (STSMs) and the generation of scientific literature. Overall, the Action has attained all of the proposed objectives and has made considerable difference by putting proteomics on the global map for animal and veterinary researchers in general and by contributing significantly to reduce the East-West and North-South gaps existing in the European farm animal research. Future activities of significance in the field of scientific research, involving members of the action, as well as others, will likely be established in the future.


Assuntos
Criação de Animais Domésticos , Tecnologia de Alimentos , Proteoma , Proteômica , Criação de Animais Domésticos/tendências , Fenômenos Fisiológicos da Nutrição Animal , Bem-Estar do Animal , Animais , Animais Domésticos , Aquicultura , Argentina , Austrália , Laticínios , Europa (Continente) , União Europeia , Tecnologia de Alimentos/tendências , Israel , Carne , Nova Zelândia , Proteômica/tendências
15.
Br J Pharmacol ; 105(2): 367-75, 1992 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-1348444

RESUMO

1. Electrical and mechanical responses to field stimulation (1-64 Hz, 0.5 ms supramaximal voltage) were recorded simultaneously in the rabbit saphenous artery. The electrical response consisted entirely of excitatory junction potentials (e.j.ps) which were abolished by alpha, beta methylene ATP (alpha, beta MeATP, 10(-6) M) and by tetrodotoxin (TTX, 10(-6) M) but were unaffected by the alpha 1-adrenoceptor antagonist, prazosin (10(-6) M). No additional electrical response was evoked by field stimulation, even in the presence of normetanephrine (NMN) and desmethylimipramine (DMI, each 10(-6) M), which block neuronal and extraneuronal uptake of noradrenaline (NA) respectively. Action potentials to field stimulation were produced only in the presence of tetraethylammonium (10(-3) M) which also enhanced the contraction. 2. Contractions to field stimulation were reduced (by some 50%) by prazosin (10(-6) M) and abolished by the additional presence of alpha, beta MeATP (10(-6) M), which blocks purinoceptors by desensitization, suggesting the involvement of both NA and an ATP-like substance in the contractile response. 3. Idazoxan (10(-6) M) which blocks prejunctional alpha 2-adrenoceptors, significantly increased the amplitude of both e.j.ps and the contraction to field stimulation (10 pulses, 1-4 Hz, 0.5 ms, supramaximal voltage). 4. NA (10(-2) M by pressure ejection) did not alter membrane potential even in the presence of NMN and DMI (each 10(-6) M). ATP (10(-2) M by pressure ejection) produced a concentration-dependent, alpha, beta MeATP-sensitive depolarization. 5. In tissues desensitized by constant infusion of alpha, beta MeATP (10(-6) M contractions to NA (10(-7) - 3 x 10(-5) M), histamine (10(-7) - 3 x 10(-5) M) and KCl (1-1.6 x 10(-2) M) were unaffected.6. Following restoration of the membrane potential, after an initial depolarization, alpha,beta MeATP (4 x 10-6M) did not change the amplitude of electrotonic hyperpolarizing current pulses significantly but abolished evoked ej.ps. The rates of recovery of evoked ej.ps and the depolarization to ATP (10-2M by pressure ejection) following desensitization to alpha,beta MeATP (10- 6 M) were comparable. These results suggest that the effects of a,fl MeATP are mediated selectively via receptors (purinoceptors).7. Suramin (10-3M) abolished ej.ps and the prazosin (10-6M), insenstive component of the contractile response and antagonized contractions to xfl MeATP (10-7_1i-5M), ATP (10-5_1i-3M), histamine (10- 7-3 x 10-SM) and 5-hydroxytryptamine (10-7-10-SM) but those to NA (10-7-10-SM) and KCI (1-1.6 x 10-2 M) were unaffected. Suramin is insufficiently selective, under these conditions, as a purinoceptor antagonist.


Assuntos
Músculo Liso Vascular/fisiologia , Trifosfato de Adenosina/análogos & derivados , Trifosfato de Adenosina/farmacologia , Antagonistas Adrenérgicos alfa/farmacologia , Animais , Artérias/efeitos dos fármacos , Artérias/fisiologia , Dioxanos/farmacologia , Estimulação Elétrica , Eletrofisiologia , Potenciais Evocados/efeitos dos fármacos , Idazoxano , Técnicas In Vitro , Masculino , Potenciais da Membrana/efeitos dos fármacos , Contração Muscular/efeitos dos fármacos , Músculo Liso Vascular/efeitos dos fármacos , Junção Neuromuscular/efeitos dos fármacos , Norepinefrina/farmacologia , Fosfolipídeos/metabolismo , Prazosina/antagonistas & inibidores , Prazosina/farmacologia , Coelhos , Suramina/farmacologia , Sistema Nervoso Simpático/efeitos dos fármacos , Sistema Nervoso Simpático/fisiologia
16.
Br J Pharmacol ; 106(4): 865-70, 1992 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-1327389

RESUMO

1. The effects of noradrenaline and alpha,beta,methylene adenosine 5'-triphosphate (alpha,beta,methylene ATP) on polyphosphoinositide metabolism, phosphatidylcholine hydrolysis and contraction in rabbit saphenous arteries were investigated. The effect of noradrenaline upon polyphosphoinositide metabolism was also investigated in the rat tail artery. 2. Noradrenaline (10(-7)-10(-4) M) evoked a concentration-dependent increase in total inositol phosphate accumulation in the rat tail but not in the rabbit saphenous artery. Propranolol (3 x 10(-6) M) did not alter this result in the rabbit saphenous artery. In addition, alpha,beta,methylene ATP (10(-6) M) significantly increased total inositol phosphate accumulation in the rabbit saphenous artery, while potassium chloride (8 x 10(-2) M) was ineffective. 3. Phorbol 1,2-myristate 1,3-acetate (3 x 10(-8) M) enhanced noradrenaline (10(-2)-10(-4) M)-evoked contractions in rabbit saphenous artery. The contractile responses to potassium chloride (1- 16 x 10(-2) M) in tissues treated with 6-hydroxydopamine (5 x 10(-4) M), in vitro, were unaffected by these concentrations of the phorbol ester. 4. Noradrenaline (10(-6)-10(-4) M) evoked a concentration-dependent increase in the levels of choline and choline phosphate, but not in those of glycerophosphocholine, in the rabbit saphenous artery. Choline levels increased significantly over the first 15-30 s then declined to control levels within 2 min of addition of noradrenaline (10(-5) M). A smaller initial rise in choline phosphate levels (15-30 s) was followed by a larger secondary rise at 2-4 min.5. alpha, beta, methylene ATP (10-1_ 0-4 M) also evoked a concentration-dependent increase in the levels of both choline and choline phosphate, but not those of glycerophosphocholine, in the rabbit saphenous artery. alpha, beta, methylene ATP (10-4 M) significantly increased levels of both of these products within the first 15-30 s of addition of the drug; these levels reached a stable plateau 1 min after addition.6. The maximum accumulation of choline or choline phosphate evoked by either noradrenaline or alpha, beta, methylene ATP, acting alone or in combination, was not significantly different. No evidence of synergism between noradrenaline and alpha, beta, methylene ATP was observed.7. This study demonstrates that each of the co-transmitters in the rabbit saphenous artery, noradrenaline and adenosine 5'-triphosphate (ATP), promote phosphatidylcholine hydrolysis. Noradrenaline seems to rely on phosphatidylcholine hydrolysis to mediate its contractile effects, whilst ATP promotes both polyphosphoinositide and phosphatidylcholine metabolism suggesting that multiple signal-transduction mechanisms are involved in stimulus-contraction coupling in this artery.


Assuntos
Trifosfato de Adenosina/farmacologia , Músculo Liso Vascular/efeitos dos fármacos , Norepinefrina/farmacologia , Fosfatidilcolinas/metabolismo , Fosfatidilinositóis/metabolismo , Animais , Colina/análise , Relação Dose-Resposta a Droga , Hidrólise , Técnicas In Vitro , Masculino , Músculo Liso Vascular/metabolismo , Ésteres de Forbol/farmacologia , Fosforilcolina/análise , Coelhos , Ratos , Ratos Endogâmicos , Fatores de Tempo , Trítio
17.
Br J Pharmacol ; 113(4): 1328-32, 1994 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-7889288

RESUMO

1. Contractions in human bronchial rings evoked by methacholine (10(-6) M) were reversed by single contractions of alpha-human atrial natriuretic peptide (10(-6) M), salbutamol (10(-6) M), sodium nitroprusside (10(-6) M) or isosorbide dinitrate (4.2 x 10(-5) M) and the extent of the relaxations compared. The activity of combinations of ANP with salbutamol, sodium nitroprusside and isosorbide dinitrate were compared with those for each agonist alone. 2. ANP and salbutamol were equipotent in reversing methacholine-evoked contraction and, in combination these agonists evoked an additive response. ANP and sodium nitroprusside also evoked similar degrees of relaxation and were additive, as were ANP and isosorbide dinitrate; however, with isosorbide dinitrate a higher concentration was required to evoke the same degree of relaxation as ANP, sodium nitroprusside or salbutamol. 3. Cumulative concentration-response curves to methacholine (10(-9)-3 x 10(-4) M) were examined in the presence and absence of the above bronchodilator substances, alone and in combination allowing their abilities to protect against contraction to be compared. ANP (10(-6) M) and salbutamol (10(-6) M) each attenuated subsequent contractions evoked by methacholine, an ability not shared with sodium nitroprusside (10(-6) M) or isosorbide dinitrate (4.2 x 10(-5) M). Indeed at lower concentrations of methacholine (< 3 x 10(-7) M), sodium nitroprusside evoked a paradoxical enhancement of methacholine-evoked contractions. 4. In combination, ANP and salbutamol attenuated contractions evoked by methacholine to a significantly greater degree than that seen with either agonist alone, whilst a combination of ANP and sodium nitroprusside evoked no greater effect than that seen with ANP alone. By contrast, isosorbide dinitrate and ANP together evoked a greater inhibition than ANP alone.5 These results suggest that a combination of agents such as ANP and salbutamol evokes a greater effect than either alone, both in reversing and protecting against methacholine-evoked contractions.Such combinations may be of benefit in the treatment of patients, allowing lower doses of drug to be used. Combinations of ANP and isosorbide dinitrate may likewise be of interest; however, the mechanism underlying the enhancement of ANP responses by isosorbide dinitrate requires further study.


Assuntos
Albuterol/farmacologia , Fator Natriurético Atrial/farmacologia , Brônquios/efeitos dos fármacos , Dinitrato de Isossorbida/farmacologia , Músculo Liso/efeitos dos fármacos , Nitroprussiato/farmacologia , Broncodilatadores/farmacologia , Sinergismo Farmacológico , Humanos , Técnicas In Vitro , Cloreto de Metacolina/antagonistas & inibidores , Cloreto de Metacolina/farmacologia , Contração Muscular/efeitos dos fármacos
18.
Br J Pharmacol ; 114(7): 1391-6, 1995 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-7606343

RESUMO

1. This study examined the activity and mechanisms of action of urodilatin in bovine bronchi. For comparison, the ability of urodilatin to evoke bronchodilatation or protect against subsequent challenge was compared to that of the closely related peptide alpha-human atrial natriuretic peptide (ANP). 2. Urodilatin reversed methacholine-evoked contraction in a concentration-dependent manner in bovine bronchi. In the absence of any attempt to prevent degradation by neutral endopeptidases, urodilatin was more potent than ANP in this tissue. 3. The bronchodilator properties of urodilatin were significantly augmented by the neutral endopeptidase inhibitor, phosphoramidon (3.68 x 10(-5) M). This provides evidence for at least partial degradation of urodilatin by neutral endopeptidases. With phosphoramidon present, urodilatin and ANP were equipotent. 4. In the presence of phosphoramidon (3.68 x 10(-5) M), pre-incubation with urodilatin (10(-6) M) had a protective effect against subsequent methacholine-induced contraction. This action of urodilatin was quantitatively similar to that of ANP in the presence of this endopeptidase inhibitor. 5. The actions of urodilatin appear to involve ATP-sensitive K+ channels since tolbutamide (10(-6) - 10(-5) M) significantly attenuated the relaxations induced by this peptide. 6. Small conductance Ca(2+)-activated K+ channels seem likewise to be implicated in the actions of urodilatin since blockade of these channels with apamin (10(-7) - 10(-6) M) resulted in a marked attenuation of urodilatin-evoked responses. 7. The presence of charybdotoxin (10-9 M-10-M) had no significant effect on subsequent responses tourodilatin suggesting that large conductance Ca2+-activated K+ channels are not involved in the relaxations evoked by this peptide.8. In the presence of phosphoramidon (3.68 x 10-5 M), urodilatin (10-6 M) evoked elevation of cyclic GMP levels within bovine bronchial tissue. Levels of cyclic GMP increased significantly within 5-10 s in response to this peptide and preceded the initiation of relaxant responses. Maximum increases in cyclic GMP levels were reached within 5 min; the time required for maximal relaxation evoked by this peptide.9. In conclusion, urodilatin, like ANP reversed and protected against, subsequent methacholine-induced bronchoconstriction; an action enhanced by the presence of phosphoramidon (3.68 x 1O-5 M).Associated with these actions of urodilatin was a rise in cyclic GMP levels as well as the opening of ATP-sensitive K+ and small conductance Ca2+-activated K+ channels.


Assuntos
Fator Natriurético Atrial/farmacologia , Brônquios/efeitos dos fármacos , Fragmentos de Peptídeos/farmacologia , Animais , Apamina/farmacologia , Bovinos , GMP Cíclico/metabolismo , Relação Dose-Resposta a Droga , Glicopeptídeos/farmacologia , Técnicas In Vitro , Cloreto de Metacolina/farmacologia , Canais de Potássio/efeitos dos fármacos , Canais de Potássio/metabolismo , Tolbutamida/farmacologia
19.
Br J Pharmacol ; 111(4): 1163-9, 1994 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-8032603

RESUMO

1. In this study, mechanical responses to endothelin-1 and endothelin-3 were examined in bovine bronchial smooth muscle. In addition, the involvement of phosphatidylinositol 4,5-bisphosphate hydrolysis (PIP2) in the responses to these peptides was assessed by measurement of inositol (1,4,5) trisphosphate (I(1,4,5)P3) production using a specific mass assay. 2. ET-1 evoked contractions of bovine bronchi which were concentration-dependent and initiated at between 10(-9) M and 10(-8) M. ET-1-evoked responses were unaffected by slight elevation of tone with potassium chloride (3 x 10(-2) M), methacholine (10(-6) M) or U46619 (10(-7) M). 3. Contractions to ET-1 were not altered by pre-incubation with atropine (10(-5) M), indomethacin (10(-5) M), nifedipine (10(-5) M), phosphoramidon (3.67 x 10(-5) M) or by removal of the epithelium. 4. ET-3 evoked small contractions which were not concentration-dependent. In the presence of phosphoramidon (3.67 x 10(-5) M) however, concentration-dependent contractions were obtained to ET-3 which were unaffected by atropine (10(-5) M) or by removal of the epithelium, but were significantly attenuated by indomethacin (10(-5) M). Nifedipine (10(-5) M) virtually abolished this response. 5. Both ET-1 and ET-3 (in the presence of phosphoramidon)-evoked contractions were significantly enhanced by the presence of the phorbol ester phorbol 12,13-dibutyrate (10(-8) M). Neither ET-1-, nor ET-3-mediated responses were antagonized by the protein kinase C (PKC) inhibitor, Ro 31-8220 (3 x 10(-9) - 3 x 10(-8) M). 6. ET-1 (3 x 10(-7) M) evoked a biphasic rise in levels of I(1,4,5)P3 which was unaltered by preincubation with atropine, whilst ET-3 (10(-10) - 3 x 10(-7) M) failed to alter levels of I(1,4,5)P3 at any time point examined, even in the presence of phosphoramidon (3.67 x 10(-5) M). 7. These results suggest that, in bovine bronchial smooth muscle, ET-l does not evoke contraction via cyclo-oxygenase metabolites, does not evoke release of the neurotransmitter substance acetylcholine, or require calcium influx via dihydropyridine-sensitive channels. ET-1 evokes 1(1,4,5)P3 production, but stimulation of protein kinase C may not be critical for the associated contraction. In contrast,ET-3-evoked contractions are partly mediated by cyclo-oxygenase metabolites. ET-3 does not stimulate PIP2 hydrolysis, nor activate PKC, but may, either directly or as a requirement of intermediates released in response to ET-3, rely upon extracellular calcium.


Assuntos
Brônquios/efeitos dos fármacos , Endotelinas/farmacologia , Músculo Liso/efeitos dos fármacos , Animais , Brônquios/fisiologia , Bovinos , Relação Dose-Resposta a Droga , Técnicas In Vitro , Inositol 1,4,5-Trifosfato/metabolismo , Contração Muscular/efeitos dos fármacos , Músculo Liso/fisiologia
20.
Eur J Pharmacol ; 385(1): 29-37, 1999 Nov 26.
Artigo em Inglês | MEDLINE | ID: mdl-10594342

RESUMO

We investigated the effect of chronic hypoxia (10% O(2) for 14 days) on airway responsiveness in rats. Chronic hypoxia significantly (P<0. 05, P<0.01, P<0.01, respectively) attenuated contractions evoked by methacholine (10(-9)-3x10(-4) M), endothelin-1 (10(-10)-3x10(-7) M) and potassium chloride (10(-3)-7x10(-2) M) in rat isolated trachea. To investigate this attenuation, we studied the effect of epithelial removal, indomethacin (3x10(-6) M), and L-nitro arginine methyl ester (L-NAME, 10(-4) M), on contractile responses in control and chronically hypoxic rat trachea. Indomethacin did not alter contractions evoked by methacholine or endothelin-1 in control or hypoxic rats. In contrast, epithelial removal and L-NAME both significantly potentiated responses to methacholine and endothelin-1 in trachea from control and chronically hypoxic rats. In separate experiments, tracheal rings were first contracted with methacholine (10(-6) M) and then relaxed, either by the nitric oxide donor sodium nitroprusside or by the beta(2)-adrenoceptor agonist, salbutamol. Sodium nitroprusside was significantly (P<0.001) more effective at reversing induced tone in tracheal rings from chronically hypoxic than control rats. Salbutamol, however, was equally effective in chronically hypoxic and control rats. These results suggest that, in trachea from both control and chronically hypoxic rats, contractile responses to methacholine and endothelin-1 are inhibited by nitric oxide, probably released from the epithelium. The attenuation of contractile responses in airways from chronically hypoxic rats may be due to an enhanced guanylyl cyclase activity and hence, an increased response to nitric oxide.


Assuntos
Hipóxia/fisiopatologia , Contração Muscular/fisiologia , Traqueia/fisiopatologia , Albuterol/farmacologia , Animais , Brônquios/efeitos dos fármacos , Brônquios/fisiopatologia , Broncoconstritores/farmacologia , Broncodilatadores/farmacologia , Inibidores de Ciclo-Oxigenase/farmacologia , Relação Dose-Resposta a Droga , Endotelina-1/farmacologia , Inibidores Enzimáticos/farmacologia , Epitélio/fisiologia , Técnicas In Vitro , Indometacina/farmacologia , Masculino , Cloreto de Metacolina/farmacologia , Contração Muscular/efeitos dos fármacos , Relaxamento Muscular/efeitos dos fármacos , NG-Nitroarginina Metil Éster/farmacologia , Óxido Nítrico/fisiologia , Óxido Nítrico Sintase/antagonistas & inibidores , Nitroprussiato/farmacologia , Cloreto de Potássio/farmacologia , Ratos , Ratos Wistar , Traqueia/efeitos dos fármacos
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