RESUMEN
AIMS: To describe basic husbandry (veterinary care, substrate and bedding, toys, frequency of cleaning) provided to guinea pigs (Cavia porcellus) by a sample of owners in New Zealand. METHODS: Respondents were invited to complete a survey disseminated through the social media channels of animal interest and advocacy groups (3 September 2020 to 3 November 2020). Respondents who agreed to participate were asked a range of questions regarding the provision of husbandry to their oldest pet guinea pig. Descriptive statistics are reported here relating to husbandry, length of ownership, provision of substrate, bedding and toys, frequency of cleaning and veterinary care. RESULTS: A total of 503 responses were received, of which 329 complete responses were analysed. Of these respondents, 208/329 (63.2%) had owned guinea pigs for more than 2 years. Most owners provided a cage with a wooden base as substrate (144/321, 44.9%), bedding (308/329; 93.6%) and toys (169/329; 51.1%). Half (176/329; 53.5%) of respondents reported taking their guinea pig to a veterinarian. Just over half of the owners surveyed cleaned water (165/329; 50.1%) and food (181/329; 55.0%) bowls daily, and a third picked up droppings (109/329; 33.1%) daily. CONCLUSIONS AND CLINICAL RELEVANCE: While owners provided an array of toys, and a range of bedding and substrate types to their guinea pigs, cages were cleaned out less frequently than recommended, and it was common for guinea pig cages to be cleaned out less frequently than recommended. Future research is required to provide robust and evidence-based links between husbandry and the welfare of pet guinea pigs.
Asunto(s)
Crianza de Animales Domésticos , Cobayas , Propiedad , Animales , Humanos , Estudios Transversales , Nueva Zelanda , Propiedad/estadística & datos numéricos , Encuestas y Cuestionarios , Veterinarios , Crianza de Animales Domésticos/normas , Crianza de Animales Domésticos/estadística & datos numéricos , Bienestar del AnimalRESUMEN
AIMS: To describe the size and type of housing used by owners for pet guinea pigs (Cavia porcellus) in New Zealand. METHODS: A survey was distributed via social media (3 September 2020 to 3 November 2020) to guinea pig interest groups and animal agencies in New Zealand. Participation was self-selected by respondents who were asked a range of questions regarding the housing and husbandry of their oldest guinea pig. Data regarding conspecifics, cage location and type, time spent out of the enclosure and size are reported here. Descriptive statistics were calculated and the associations between housing type and size were assessed. RESULTS: A sample of 330 owners provided details of housing for their guinea pig. Most respondents housed their guinea pigs in groups of two or more (283/330; 85.7%). The most frequently reported housing types were a hutch or cage with an attached run (155/330; 47.0%) and inside the house in their own area (90/330, 27.3%). The mean size of enclosures was 3.3 (SD 4.3; median 2.0; min 0.3: max 37.5) m2 and the mean area provided per guinea pig was 1.4 (SD 1.6; median 0.9; min 0.2; max 10) m2. Of the owners using enclosures, 59/284, (21.1%) provided less cage space than the minimum recommended by the Royal New Zealand Society for the Prevention of Cruelty to Animals (RNZSPCA; 1.0â m2 per pair of guinea pigs). Nearly two-thirds (89/318; 59.4%) of respondents provided their guinea pigs daily time in a different area to their main living area (pen, garden, house, deck or garage) on a daily basis. CONCLUSIONS AND CLINICAL RELEVANCE: While mean cage size reported by respondents was greater than that recommended for pet guinea pigs by the RNZSPCA and fell within the range recommended by animal welfare groups internationally, a notable proportion of guinea pigs were housed in cages smaller than the recommended size. Furthermore the majority of guinea pigs were not provided with time outside this cage. Further work to investigate the effect of housing size on welfare of pet guinea pigs is required.
Asunto(s)
Bienestar del Animal , Vivienda para Animales , Animales , Estudios Transversales , Cobayas , Humanos , Nueva Zelanda , Encuestas y CuestionariosRESUMEN
Adenosine diphosphate (ADP) is an important platelet agonist and ADP released from platelet dense granules amplifies responses to other agonists. There are three known subtypes of ADP receptor on platelets: P2X(1), P2Y(1) and P(2T) receptors. Sustained ADP-induced aggregation requires co-activation of P2Y(1) and P(2T) receptors. AR-C69931MX, a selective P(2T) receptor antagonist and novel antithrombotic agent, was studied to characterize further the function of the P(2T) receptor. The roles of the P2Y(1) receptor and thromboxane A(2) were assessed using the selective P2Y(1) antagonist A2P5P and aspirin respectively. Aggregation was measured by whole blood single-platelet counting and platelet-rich plasma turbidimetry, using hirudin anticoagulation. Dense granule release was estimated using ([14)C]-5-hydroxytryptamine (HT)-labelled platelets. Ca(2+) mobilization, P-selectin expression, Annexin V binding and microparticle formation were determined by flow cytometry. P(2T) receptor activation amplified ADP-induced aggregation initiated by the P2Y(1) receptor, as well as amplifying aggregation, secretion and procoagulant responses induced by other agonists, including U46619, thrombin receptor-activating peptide (TRAP) and collagen, independent of thromboxane A(2) synthesis, which played a more peripheral role. P(2T) receptor activation sustained elevated cytosolic Ca(2+) induced by other pathways. These studies indicate that the P(2T) receptor plays a central role in amplifying platelet responses and demonstrate the clinical potential of P(2T) receptor antagonists.
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Adenosina Monofosfato/análogos & derivados , Fibrinolíticos/farmacología , Proteínas de la Membrana , Activación Plaquetaria/efectos de los fármacos , Agregación Plaquetaria/efectos de los fármacos , Receptores Purinérgicos P2/fisiología , Ácido 15-Hidroxi-11 alfa,9 alfa-(epoximetano)prosta-5,13-dienoico/farmacología , Adenosina Difosfato/metabolismo , Adenosina Monofosfato/farmacología , Análisis de Varianza , Anexina A5/metabolismo , Aspirina/farmacología , Plaquetas/metabolismo , Calcio/metabolismo , Colágeno/farmacología , Citosol/metabolismo , Citometría de Flujo , Humanos , Selectina-P/metabolismo , Fragmentos de Péptidos/farmacología , Antagonistas del Receptor Purinérgico P2 , Receptores Purinérgicos P2/efectos de los fármacos , Receptores Purinérgicos P2X , Receptores Purinérgicos P2Y1 , Receptores Purinérgicos P2Y12 , Serotonina/metabolismo , Estimulación Química , Tromboxano A2/agonistas , Tromboxano A2/metabolismoRESUMEN
We have shown previously that a 500-bp region of the human insulin receptor promoter (-0.3 to -1.8 kb) was able to stimulate transcription from a heterologous thymidine kinase promoter in HepG2 hepatoma cells but not in HeLa fibroblasts. Footprint analysis localized the transcription factor binding sites to a 36-bp region at -1420. In this paper, we analyze the factors that recognize this element and show that it contains binding sites for the CAAT/enhancer binding protein C/EBP and nuclear factor 1 (NF-1). In addition we show that both C/EBP alpha and the C/EBP beta can transactivate the human insulin receptor promoter in a dose-dependent manner.
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Proteínas de Unión al ADN/metabolismo , ADN/metabolismo , Genes Reguladores , Proteínas Nucleares/metabolismo , Regiones Promotoras Genéticas , Receptor de Insulina/genética , Factores de Transcripción/metabolismo , Secuencia de Bases , Sitios de Unión , Proteínas Potenciadoras de Unión a CCAAT , Carcinoma Hepatocelular , Cloranfenicol O-Acetiltransferasa/biosíntesis , Cloranfenicol O-Acetiltransferasa/metabolismo , Células HeLa , Humanos , Neoplasias Hepáticas , Datos de Secuencia Molecular , Factores de Transcripción NFI , Oligodesoxirribonucleótidos , Oligonucleótidos Antisentido , Timidina Quinasa/biosíntesis , Timidina Quinasa/genética , Transcripción Genética , Transfección , Células Tumorales Cultivadas , Proteína 1 de Unión a la Caja YRESUMEN
The ability of the human insulin receptor promoter to direct expression of a linked chloramphenicol acetyltransferase gene was assessed in transient transfections into HepG2 and Hela cells. A 5'-deletional analysis of the promoter showed that regions between -646 and -489 were important for the activity of the proximal promoter. In addition, a possible negative regulatory element was identified between -1311 and -877 and a positive element between -1823 and -1311. DNase I footprint and gel retardation analysis showed that multiple factors bind to the human insulin receptor promoter. In particular, DNase I protection patterns were observed over the Sp1 sites at -620 to -599 and -438 to -392, a TC box at -533, four homopyrimidine/homopurine sites clustered around -1150, and a site at -1420 that contains the motif TGGCCC which has been shown to bind the liver-specific transcription factor LF-A1.
Asunto(s)
Regulación de la Expresión Génica , Proteínas Nucleares , Regiones Promotoras Genéticas , Receptor de Insulina/genética , Factores de Transcripción/metabolismo , Transcripción Genética , Secuencia de Bases , Sitios de Unión , Carcinoma Hepatocelular , Línea Celular , Cloranfenicol O-Acetiltransferasa/biosíntesis , Cloranfenicol O-Acetiltransferasa/genética , Deleción Cromosómica , Clonación Molecular , Proteínas de Unión al ADN/metabolismo , Desoxirribonucleasa I , Células HeLa , Factor Nuclear 1 del Hepatocito , Factor Nuclear 1-alfa del Hepatocito , Factor Nuclear 1-beta del Hepatocito , Humanos , Neoplasias Hepáticas , Datos de Secuencia Molecular , Plásmidos , Receptor de Insulina/biosíntesis , Proteínas Recombinantes de Fusión/biosíntesis , Mapeo Restrictivo , TransfecciónRESUMEN
We have used a tris(4-methoxyphenyl)-phosphonium compound as a hapten to elicit catalytic antibodies that selectively remove trityl protecting groups at neutral pH. One antibody, 37C4, was characterized kinetically with a number of trityl substrates. The rate enhancement was consistently near 200; the Km was approximately 30 microM for the methoxytrityl substrates. Compounds with no methoxy substituents on the trityl group were not hydrolysed by the antibody. No decrease in the rate of reaction was detected through 21 turnovers, which suggests that the presumptive trityl cation formed during the cleavage reaction does not alkylate the antibody binding pocket. The rates of the background and antibody-catalysed reactions both increase logarithmically with decreasing pH, implying that general acid catalysis is not involved: further studies will test this assumption. The favoured mechanism for the catalytic activity of antibody 37C4 is charge complementarity in the binding site stabilizing a positively charged intermediate(s) in the cleavage reaction. The coding sequence for 37C4 is being cloned into a phage lambda vector in preparation for site-directed mutagenesis to improve the catalytic efficiency of the antibody.