RESUMO
AIMS: To examine and assess causes of mortality of kiwi (Apteryx spp.) submitted to Massey University between 2010 and 2020 across the five recognised species according to location, age group and captivity status in New Zealand. METHODS: Post-mortem reports were obtained from the Massey University/Te Kunenga ki Purehuroa School of Veterinary Science/Wildbase Pathology Register. Inclusion criteria were all species of kiwi with a date of post-mortem examination between August 2010 and August 2020. Data from each report was exported, categorised and compared using Microsoft Excel. RESULTS: Of a total of 1,005 post-mortem reports, there were 766 North Island brown kiwi (NIBK; A. mantelli), 83 tokoeka (A. australis), 73 rowi (A. rowi), 49 great spotted kiwi (A. haastii), and 34 little spotted kiwi (A. owenii). This comprised 19 eggs/embryos, 125 neonatal, 473 juvenile, 153 subadult, and 235 adult kiwi. There were 615 kiwi from wild populations, 148 from sanctuary populations, 238 from captivity, and four from unspecified locations. The leading cause of death was trauma, affecting 322 (32.0 (95% CI = 29.2-35.0)%) kiwi including 289 (37.3 (95% CI = 26.0-31.7)%) NIBK. Nearly half of these died from predation by mustelids, with losses recorded from neonates to adults and clustered in the central to southern North Island. Predation by dogs was the second most common cause of death, killing 84 (8.4 (95% CI = 6.7-10.2)%) kiwi, of which 65.5% came from the northern districts of the North Island. Non-infectious disease killed 214 (21 (95% CI = 18.8-24.0)%) kiwi, and included developmental deformities, gastrointestinal foreign bodies and predator trap injuries. Infectious disease killed 181 (18.0 (95% CI = 15.7-20.5)%) kiwi and the proportion decreased with age, with common diagnoses including coccidiosis, bacterial septicaemia, avian malaria, and fungal respiratory disease. Starvation affected 42 (4.2 (95% CI = 3.0-5.6)%) kiwi, comprised of mainly neonatal or juvenile individuals from wild or sanctuary populations, with a higher percentage seen in tokoeka (11/83; 13.3%) compared to other species (min 0%, max 5.9%). The cause of death was undetermined in 246 (24.5 (95% CI = 21.8-27.3)%) cases, which was most often due to poor preservation of remains. This included 33/73 (46%) rowi and 32/83 (39%) tokoeka, and affected mainly birds from sanctuary and wild populations. CONCLUSIONS: This study enhances our understanding of causes of mortality in captive, wild and sanctuary populations of all kiwi species and age groups within contemporary New Zealand.
Assuntos
Doenças das Aves , Doenças do Cão , Paleógnatas , Animais , Cães , Doenças das Aves/microbiologia , Nova Zelândia/epidemiologia , Estudos Retrospectivos , Autopsia/veterinária , ÓvuloRESUMO
AIMS: To investigate the pathogenesis of a disease in takahe (Porphyrio hochstetteri) with intracytoplasmic inclusion bodies in lower motor neurons. METHODS: Four birds aged between 5 and 12 years, from three different wildlife sanctuaries in New Zealand were examined. Of these, only one had signs of spinal dysfunction in the form of paresis. Stained paraffin sections of tissues were examined by light microscopy and immunostained sections of the ventral horn of the spinal cord by confocal microscopy. Epoxy resin sections of the spinal cord from the bird with spinal dysfunction were examined by electron microscopy. RESULTS: Two types of inclusion bodies were noted, but only in motor neurons of the ventral spinal cord and brain stem. These were large globoid eosinophilic bodies up to 5â µm in diameter, and yellow/brown granular inclusions mostly at the pole of the cell. The globoid bodies stained with Luxol fast blue but not with periodic acid Schiff (PAS), or Sudan black. The granular inclusions stained with Luxol fast blue, PAS and Sudan black. Both bodies were slightly autofluorescent. On electron microscopy the globoid bodies had an even electron-dense texture and were bound by a membrane. Beneath the membrane were large numbers of small intraluminal vesicles. The smaller granular bodies were more heterogeneous, irregularly rounded and membrane-bound accumulations of granular electron-dense material, often with electron-lucent vacuoles. Others were more vesicular but contained varying amounts of electron-dense material. The large globoid bodies did not immunostain for lysosomal markers lysosomal associated protein 1 (LAMP1) or cathepsin D, so were not lysosomal. The small granular bodies stained for cathepsin D by a chromogenic method.A kindred matrix analysis showed two cases to be as closely related as first cousins, and another case was almost as closely related to one of them, but the fourth bird was unrelated to any other. CONCLUSIONS: It was concluded that this was an endoplasmic reticulum storage disease due to a specific protein misfolding within endoplasmic reticulum. It was rationalised that the two types of inclusions reflected the same aetiology, but that misfolded protein in the smaller granular bodies had entered the lysosomal system via endoplasmic reticulum autophagy. Although the cause was unclear, it most likely had a genetic aetiology or predisposition and, as such, has clinical relevance.
Assuntos
Catepsina D , Doença dos Neurônios Motores , Animais , Catepsina D/metabolismo , Retículo Endoplasmático/metabolismo , Retículo Endoplasmático/patologia , Doença dos Neurônios Motores/veterinária , Doença dos Neurônios Motores/metabolismo , Doença dos Neurônios Motores/patologia , Microscopia Eletrônica/veterinária , AvesRESUMO
CASE HISTORY: A 1-year-old female New Zealand sea lion (Phocarctos hookeri) was intermittently observed in the Otago region of New Zealand over an 11-month period, always dragging her hind flippers. In December 2012 the sea lion was found dead, after a period of several days being observed to be harassed by male sea lions. PATHOLOGICAL FINDINGS: At gross postmortem examination the sea lion was in moderate body condition with signs of recent bite wounds and bruising. The lungs were dark and poorly inflated. Histological findings included meningoencephalomyelitis, radiculomyelitis of the cauda equina, myocarditis and myositis. Toxoplasmosis gondii organisms were detected histologically and following immunohistochemistry in the brain, spinal cord, spinal nerves and pelvic muscles. MOLECULAR BIOLOGY: Nested PCR analysis and sequencing confirmed the presence of T. gondii DNA in uterine and lung tissue. A variant type II T. gondii genotype was identified using multilocus PCR-restriction fragment length polymorphism analysis. DIAGNOSIS: Systemic toxoplasmosis. CLINICAL RELEVANCE: Infection with T. gondii involving the spinal cord and nerves was the likely cause of the paresis observed in this sea lion before death. Ultimately, death was attributed to crushing and asphyxiation by a male sea lion, presumably predisposed by impaired mobility. Diagnosis of toxoplasmosis in a New Zealand sea lion highlights the possibility that this disease could play a role in morbidity and mortality in this endangered species, particularly in the recently established mainland populations that are close to feline sources of T. gondii oocysts.
Assuntos
Leões-Marinhos/parasitologia , Toxoplasmose Animal/epidemiologia , Animais , Feminino , Nova Zelândia/epidemiologia , Reação em Cadeia da Polimerase/veterinária , Toxoplasma/genética , Toxoplasmose Animal/patologiaRESUMO
Stillbirth is a small and often cryptic fraction of neonatal mortality in mammals including pinnipeds. As part of an investigation into the poor reproductive success of the endangered New Zealand sea lion (Phocarctos hookeri), archived tissues from 37 stillborn pups born on Enderby Island between 1998 and 2012 were examined using histopathological techniques. Apart from bronchopneumonia with neutrophilic infiltration in 4 cases, few inflammatory conditions were identified in stillborn pups. However, 27/32 (84%) stillborn pups had aspirated squames present in the respiratory tract, without meconium. It is unclear if this finding represents fetal distress during parturition or whether it is a normal finding for this species. Three pups lacked histological evidence of hepatic glycogen storage, which may indicate placental defects or maternal undernutrition. No evidence of infectious disease was found on histopathological analysis, consistent with the low seroprevalence in New Zealand of infections known to cause reproductive failure in other pinniped species. This study forms an important baseline for further examination of stillborn New Zealand sea lion pups, as pup mortality is investigated as a contributor to the species' decline.
Assuntos
Leões-Marinhos , Natimorto/veterinária , Animais , Animais Recém-Nascidos , Espécies em Perigo de Extinção , Feminino , Feto/patologia , Masculino , Nova Zelândia/epidemiologia , Natimorto/epidemiologiaRESUMO
Determining the equilibrium-binding affinity (Kd) of two interacting proteins is essential not only for the biochemical study of protein signaling and function but also for the engineering of improved protein and enzyme variants. One common technique for measuring protein-binding affinities uses flow cytometry to analyze ligand binding to proteins presented on the surface of a cell. However, cell-binding assays require specific considerations to accurately quantify the binding affinity of a protein-protein interaction. Here we will cover the basic assumptions in designing a cell-based binding assay, including the relevant equations and theory behind determining binding affinities. Further, two major considerations in measuring binding affinities-time to equilibrium and ligand depletion-will be discussed. As these conditions have the potential to greatly alter the Kd, methods through which to avoid or minimize them will be provided. We then outline detailed protocols for performing direct- and competitive-binding assays against proteins displayed on the surface of yeast or mammalian cells that can be used to derive accurate Kd values. Finally, a comparison of cell-based binding assays to other types of binding assays will be presented.
Assuntos
Ligação Proteica , Mapas de Interação de Proteínas , Proteínas/química , Animais , Ligação Competitiva , Citometria de Fluxo , Humanos , Cinética , Ligantes , Mamíferos , LevedurasRESUMO
The release of arachidonic acid and its metabolites, prostaglandin E2 and thromboxane A2, from WI-38 human lung fibroblasts was modulated by p-hydroxymercuribenzoate. Exposure to the inhibitor resulted in a dose-dependent decrease in [1-14C]arachidonic acid uptake and incorporation into phospholipids and neutral lipid pools. Activities of lung fibroblast arachidonyl-CoA synthetase and lysolecithin acyltransferase were inhibited by 100 microM p-hydroxymercuribenzoate. [14C]Arachidonic acid labelled fibroblasts exhibited an increased release of [14C]arachidonate and [14C]prostaglandin E2 of 54% and 112%, respectively, when exposed to 100 microM of inhibitor. The stimulatory effects of 8.0 microM delta 1-tetrahydrocannabinol on arachidonate release and prostaglandin E synthesis (Burstein, S., Hunter, S.A., Sedor, C. and Shulman, S. (1982) Biochem. Pharmacol. 31, 2361-2365) were modified by the inclusion of inhibiting agent, resulting in a 608% stimulation in arachidonic acid release, while prostaglandin E2 and thromboxane A2 synthesis increased 894% and 390%, respectively, over levels obtained by untreated cells. The levels of arachidonate metabolites were altered by inhibitor when compared to cells treated with cannabinoid alone. No significant inhibition by delta 1-tetrahydrocannabinol was found on arachidonic uptake in these cells. In unlabelled studies, p-hydroxymercuribenzoate resulted in a profound, dose-dependent stimulation of prostaglandin E synthesis of 1490% at 150 microM inhibitor concentration. These results provide evidence that free arachidonate is reincorporated via acylation, thereby implicating this pathway as a possible control mechanism for the synthesis of arachidonic acid metabolites.
Assuntos
Hidroximercuribenzoatos/farmacologia , Pulmão/metabolismo , Fosfolipídeos/metabolismo , Prostaglandinas E/biossíntese , Tromboxano A2/biossíntese , Tromboxanos/biossíntese , 1-Acilglicerofosfocolina O-Aciltransferase/antagonistas & inibidores , Acilação , Linhagem Celular , Cloromercurobenzoatos/farmacologia , Coenzima A Ligases/antagonistas & inibidores , Meios de Cultura/análise , Dinoprostona , Dronabinol/farmacologia , Fibroblastos/metabolismo , Humanos , Ácido p-CloromercurobenzoicoRESUMO
A concentration-related stimulation of anandamide (arachidonylethanolamide) synthesis by delta 9-tetrahydrocannabinol (THC) was observed in N-18TG2 neuroblastoma cells. Anandamide was detected and measured using an approach in which [3H]arachidonic acid and [14C]ethanolamine were incorporated into the phospholipids of subconfluent monolayers of cells, and the radiolabeled products were analyzed by TLC following agonist exposure. Both precursors showed similar concentration-response relationships and time dependencies consistent with the production of a product containing both the ethanolamine and arachidonyl moieties. The radiolabeled product also migrated together with authentic anandamide on two-dimensional TLC, confirming its identity as arachidonylethanolamide. Approximately two-thirds of the observed synthesis could be inhibited by 1 microM wortmannin, an agent previously reported to inhibit THC-stimulated arachidonic acid release. These findings are in agreement with reports showing that THC can mobilize phospholipid bound arachidonic acid, leading to the production of other eicosanoids.
Assuntos
Ácidos Araquidônicos/biossíntese , Dronabinol/farmacologia , Ácido Araquidônico/metabolismo , Cromatografia em Camada Fina , Endocanabinoides , Etanolamina , Etanolaminas/metabolismo , Neuroblastoma/metabolismo , Neuroblastoma/patologia , Alcamidas Poli-Insaturadas , Células Tumorais CultivadasRESUMO
Prior exposure of cells in vitro to delta 1-tetrahydrocannabinol-7-oic acid (delta 1-THC-7-oic acid) reduced the degree of stimulation of prostaglandin synthesis incurred by subsequent treatment with delta 1-THC. The site of action of this inhibitory effect seemed to be on cyclooxygenase and not at the earlier step involving the phospholipase-mediated release of arachidonic acid. delta 1-THC-7-oic acid is a major metabolite of delta 1-THC and has no psychoactivity in humans. Our findings raise the possibility, however, that it may influence the in vivo activities of delta 1-THC by antagonizing its stimulatory action on cellular prostaglandin synthesis.
Assuntos
Dronabinol/análogos & derivados , Dronabinol/antagonistas & inibidores , Animais , Anti-Inflamatórios/farmacologia , Ácido Araquidônico , Ácidos Araquidônicos/metabolismo , Inibidores de Ciclo-Oxigenase , Dinoprostona , Dronabinol/metabolismo , Dronabinol/farmacologia , Humanos , Camundongos , Camundongos Endogâmicos , Prostaglandinas E/biossíntese , Tromboxanos/biossínteseRESUMO
The exposure of cells in culture to cannabinoids results in a rapid and significant mobilization of phospholipid bound arachidonic acid. In vivo, this effect has been observed as a rise in eicosanoid tissue levels that may account for some of the pharmacological actions of delta 9-tetrahydrocannabinol (THC), the major psychoactive cannabinoid. Fluoroaluminate pretreatment of mouse peritoneal cells potently reduced the cannabinoid response, while promoting arachidonate release on its own, consistent with earlier observations that this effect may be a receptor/G-protein-mediated process. Further support for receptor mediation was the demonstration of saturable, high-affinity cannabinoid binding in these cells. THC potency was reduced in the presence of ethanol, and was accompanied by significant increases in phosphatidylethanol (PdEt) levels, a unique product of phospholipase D (PLD) activity. THC-dependent arachidonate release was reduced partially in similar amounts by either propranolol or wortmannin, further implicating PLD as a mediator of THC action. A central role for diacylglyceride (DAG), a secondary product of PLD metabolism, in this THC-induced process, both as a source of arachidonate and as a stimulator of protein kinase C (PKC), is supported by the data in this report. Cells exposed to phorbol ester for 18 hr prior to THC challenge became less responsive, indicating a possible role for PKC. The involvement of PKC further suggests participation by phospholipase A2 (PLA2) whose activity may be regulated by the former. Treatment of cells with interleukin-1 alpha, an agent known to elevate PLA2 levels, caused an increase in the THC response, supporting a role for this enzyme in the release reaction. Direct evidence, by immunoblotting, for the activation and phosphorylation of PLA2 by THC was also obtained. In summary, the evidence presented in this report indicates that THC-induced arachidonic acid release occurs through a series of events that are consistent with a receptor-mediated process involving the stimulation of one or more phospholipases.
Assuntos
Ácido Araquidônico/metabolismo , Dronabinol/farmacologia , Fosfolipases A/metabolismo , Animais , Células Cultivadas , Relação Dose-Resposta a Droga , Dronabinol/antagonistas & inibidores , Ativação Enzimática/efeitos dos fármacos , Interleucina-1/farmacologia , Ligantes , Camundongos , Fosfolipase D/antagonistas & inibidores , Fosfolipase D/metabolismo , Fosfolipases A2 , Monoéster Fosfórico Hidrolases/antagonistas & inibidores , Receptores de Canabinoides , Receptores de Droga/metabolismoRESUMO
Preliminary data [S. Burstein and S. A. Hunter, Biochem. Pharmac. 27, 1275 (1978)] showed that cannabinoids at levels of 1 microM or greater elevated the concentrations of prostaglandins in cell culture models. Further study [S. Burstein and S. A. Hunter, J. clin. Pharmac. 21, 240S (1981)] led to the suggestion that this effect was due to a stimulation of phospholipase A2 resulting in the release of free arachidonic acid which was then partly converted into the prostaglandin(s) normally synthesized by the particular target system. The present report gives detailed data on the cannabinoid-induced synthesis of prostaglandin E2 by te WI-38 fibroblast derived from human lung. The effect could be blocked by pretreatment with mepacrine, a phospholipase inhibitor, and aspirin, a cyclooxygenase inhibitor. These findings lend support to the hypothesis that some of the in vivo actions of the cannabinoids are due to modulations in prostaglandin synthesis at various tissue sites.
Assuntos
Dronabinol/farmacologia , Pulmão/metabolismo , Prostaglandinas E/biossíntese , Ácido Araquidônico , Ácidos Araquidônicos/metabolismo , Canabidiol/farmacologia , Células Cultivadas , Dinoprostona , Fibroblastos/metabolismo , Humanos , Pulmão/efeitos dos fármacosRESUMO
The phospholipases controlling the release of arachidonic acid in mouse peritoneal macrophages have been shown to be stimulated by the natural psychoactive cannabinoids. A close correlation was observed between the potencies of these substances in elevating arachidonate levels in vitro and the reported activities in a behavioral assay in monkeys and in producing a "high" in humans. The order of activity with the macrophages was delta 1-tetrahydrocannabinol (delta 1-THC) greater than 7-OH-delta 1-THC greater than 6 alpha-OH-delta 1-THC greater than 6 beta-delta 1-THC much greater than delta 6-THC-7-oic acid. It is suggested that this effect, which has now been shown in several diverse cell types, may serve as a model for studying the mechanism of action of THC.
Assuntos
Ácidos Araquidônicos/metabolismo , Dronabinol/farmacologia , Macrófagos/metabolismo , Animais , Ácido Araquidônico , Líquido Ascítico , Dronabinol/metabolismo , Técnicas In Vitro , Lipoxigenase/análise , Masculino , Camundongos , Camundongos Endogâmicos , Relação Estrutura-AtividadeRESUMO
OBJECTIVE: To determine if compliance with annual tuberculosis skin testing correlated with the number of cases of tuberculosis seen in patients and healthcare workers. DESIGN: Survey using a written questionnaire. SETTING AND PARTICIPANTS: 159 Veterans' Administration facilities. RESULTS: Hospitals that reported that > 80% of their healthcare workers received annual skin tests saw 12.7 patient cases per 10,000 admissions and 4.0 healthcare worker cases per 10,000 personnel. Facilities in which < 20% of their healthcare workers were given annual skin tests saw 4.5 cases per 10,000 admissions and 1.6 cases in healthcare workers per 10,000 personnel (P < .001 for patients and P = .31 for healthcare workers). The ratio of the median number of patients placed in acid-fast bacilli (AFB) isolation to the median number of patients with confirmed tuberculosis was 12. There was no correlation of this ratio with the number of cases of tuberculosis in patients or healthcare workers seen in each facility. CONCLUSION: Compliance with annual tuberculosis skin testing was related directly to the rate of tuberculosis seen in patients. More standardized policies for placing patients in AFB isolation are needed to control for potentially costly variation among facilities. These measures should have highest priority in the control of tuberculosis in the healthcare setting, before implementing still more expensive interventions.
Assuntos
Infecção Hospitalar/prevenção & controle , Hospitais de Veteranos/estatística & dados numéricos , Controle de Infecções/normas , Programas de Rastreamento/normas , Exposição Ocupacional/prevenção & controle , Recursos Humanos em Hospital , Tuberculose Pulmonar/prevenção & controle , Humanos , Inquéritos e Questionários , Teste Tuberculínico , Estados UnidosRESUMO
The previously reported release of arachidonic acid by THC has now been demonstrated in murine Leydig cells and WI-38 human lung fibroblasts showing the generality of the effect. The release reaction could be antagonized by phospholipase A2 inhibitors such as quinacrine and quinine, suggesting that THC can stimulate the activity of this enzyme. Further evidence for this possibility was obtained by demonstrating the release effect on a subcellular fraction which contained the phospholipase A2 activity. The stimulation of this enzyme could have profound effects on prostaglandin synthesis and/or on the integrity of various membrane structures.
Assuntos
Canabinoides/farmacologia , Fosfolipases A/metabolismo , Fosfolipases/metabolismo , Prostaglandinas/biossíntese , Animais , Ácido Araquidônico , Ácidos Araquidônicos/metabolismo , Células Cultivadas , Dronabinol/farmacologia , Células HeLa , Humanos , Hidrólise , Células Intersticiais do Testículo/metabolismo , Masculino , Camundongos , Fosfolipases A2 , Ratos , Frações Subcelulares/enzimologiaRESUMO
An isotopic dilution procedure using specific prostaglandin E2 (PGE2) brain receptors was utilized to determine the changes in brain PGE2 levels subsequent to drug exposure. Delta-1-tetrahydrocannabinol (delta 1-THC) stimulated PGE2 synthesis resulting in increased brain concentrations when compared with vehicle treated rats and mice. Indomethacin markedly inhibited the delta 1-THC elevated rise in PGE2 levels presumably by inhibition of prostaglandin synthetase. The delta 1-THC-induced increase in PGE2 brain levels was also suppressed by i.v. administered rabbit PGE2-antiserum. This suggests that one of the sites of delta 1-THC action is extracerebral and from here a portion of the released prostaglandins are transported to the brain. These results add further support to previous data that delta 1-THC given orally results in an increase in brain PGE2 levels.
Assuntos
Química Encefálica/efeitos dos fármacos , Dinoprostona/análise , Dronabinol/farmacologia , Animais , Anticorpos/sangue , Dinoprostona/imunologia , Dinoprostona/metabolismo , Feminino , Indometacina/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos , Ratos , Ratos Endogâmicos , TrítioRESUMO
This study examined the relationships between appropriateness of readmission within two weeks of discharge and appropriateness of previous admission and discharge, bed section, type of readmission, and patient demographic, medical condition, and hospital stay characteristics. Using the Department of Veterans Affairs (VA) Patient Treatment File and medical records, 445 readmissions to a highly affiliated midwestern VA Medical Center in fiscal year 1984 were examined. Appropriateness was determined by four trained medical record abstractors using InterQual admission and discharge standards. Type of readmission was based on a pilot-tested flowchart. Appropriateness of readmission was significantly associated with that of the previous admission and discharge, with the relationship varying by admission, discharge, and readmission bed sections. Reasons for inappropriate admissions, discharges, and readmissions also varied by bed section. For the majority of inappropriate readmissions, there was clear written evidence in the medical record during the previous hospital stay that the patient was directed to return for readmission. Inappropriate readmissions were more likely than appropriate readmissions to have a primary diagnosis of neoplasm or digestive disorder. These results indicate the importance of examining both the operational efficiencies during the previous admission and the clinical criteria for admitting, discharging, and readmitting patients in assessing the appropriateness of readmissions.
Assuntos
Admissão do Paciente , Alta do Paciente , Readmissão do Paciente , Adulto , Idoso , Hospitais de Veteranos , Humanos , Tempo de Internação , Pessoa de Meia-Idade , Planejamento de Assistência ao Paciente , Fatores de Tempo , Estados UnidosRESUMO
Numerous reports have suggested that increased synthesis of eicosanoids is a significant effect of cannabinoids in several models including the human. To address the question of receptor mediation in this process we have carried out experiments using oligonucleotides that are antisense to the CB1 and to the CB2 receptors. We have synthesized sense, antisense and random oligonucleotide probes to test for receptor involvement in THC stimulation of arachidonic acid release in three cell lines of both central and peripheral origin. Treatment of N18 mouse neuroblastoma cells with the CB1 antisense probe, at two concentrations, resulted in a dramatic decrease of THC stimulated arachidonate release while treatment with antisense CB2 was less effective. Synthesis of the novel eicosanoid, anandamide, was also reduced by antisense CB1 but not by antisense CB2. Western blot analysis indicated a decreased level of CB1 in CB1 antisense treated cells. The CB1 antagonist, SR141716A, was effective in reducing the THC elevated levels of free arachidonate in these cells in agreement with the antisense data. In the macrophage line, RAW 264.7, we found that while the sense, the random and the CB1 antisense oligonucleotides were ineffective, the CB2 antisense probe gave significant reductions of the THC induced response. The CB2 probe was also effective in reducing the release of arachidonate in WI-38 human lung fibroblasts. These findings support the idea of a receptor mediated process for cannabinoid stimulation of eicosanoid synthesis.
Assuntos
Ácido Araquidônico/metabolismo , Ácidos Araquidônicos/biossíntese , Dronabinol/farmacologia , Receptores de Droga/metabolismo , Animais , Western Blotting , Linhagem Celular , Dronabinol/metabolismo , Endocanabinoides , Humanos , Camundongos , Oligonucleotídeos Antissenso/metabolismo , Oligonucleotídeos Antissenso/farmacologia , Piperidinas/farmacologia , Alcamidas Poli-Insaturadas , Pirazóis/farmacologia , Receptores de Canabinoides , Receptores de Droga/antagonistas & inibidores , Receptores de Droga/genética , Rimonabanto , Células Tumorais CultivadasRESUMO
Stereospecificity has been reported for a number of actions of the cannabinoids in a variety of systems. In the present report, we have shown that this effect can also be demonstrated when human lung fibroblasts in monolayer culture are stimulated by cannabinoids to produce prostaglandin E2 (PGE2). Three enantiomeric pairs of cannabinoids, (+) and (-)-delta 1-tetrahydrocannabinol (THC), (+) and (-)-delta 6-THC and (+) and (-)-delta 6-dimethylheptyl (DMH) THC were tested. In each case the (-) isomer was significantly more potent in agreement with the findings of others using different systems. Interestingly, very little stereospecificity was found in fibroblasts when the release of arachidonic acid, the precursor of PGE2, was monitored. This suggests that cannabinoids may act at several sites within the cell some of which show comparatively greater stereoselectivity for these agonists.
Assuntos
Canabinoides/farmacologia , Fibroblastos/metabolismo , Prostaglandinas E/biossíntese , Ácido Araquidônico , Ácidos Araquidônicos/metabolismo , Varredura Diferencial de Calorimetria , Linhagem Celular , Dinoprostona , Dronabinol/análogos & derivados , Dronabinol/farmacologia , Fibroblastos/efeitos dos fármacos , Humanos , Bicamadas Lipídicas/metabolismo , Estereoisomerismo , Relação Estrutura-AtividadeRESUMO
The release of arachidonic acid from mouse peritoneal and S49 cells induced by delta 1-tetrahydrocannabinol was found to be altered by prior exposure of the cells to either pertussis toxin or cholera toxin. The stable analogs of GTP and GDP, GTP-gamma-S and GDP-beta-S, were also effective in changing the extent of arachidonate release in saponin-treated cells. GDP-beta-S essentially abolished the THC response, while GTP-gamma-S showed effects mainly on vehicle-treated cells. The cataleptic action of THC in intact mice which is mediated by eicosanoids was also attenuated by pertussis toxin pretreatment. It is suggested that the THC receptor is coupled to phospholipases through one or more G-proteins and that adenylate cyclase probably does not have a role in this mechanism.
Assuntos
Canabinoides/farmacologia , Proteínas de Ligação ao GTP/fisiologia , Fosfolipases/metabolismo , Toxina Adenilato Ciclase , Animais , Ácido Araquidônico/metabolismo , Catalepsia/induzido quimicamente , Catalepsia/prevenção & controle , Dronabinol/antagonistas & inibidores , Dronabinol/farmacologia , Ativação Enzimática/efeitos dos fármacos , Feminino , Guanosina 5'-O-(3-Tiotrifosfato)/farmacologia , Guanosina Difosfato/análogos & derivados , Guanosina Difosfato/farmacologia , Técnicas In Vitro , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos , Toxina Pertussis , Saponinas/farmacologia , Tionucleotídeos/farmacologia , Fatores de Virulência de Bordetella/farmacologiaRESUMO
Hector's dolphins (Cephalorhynchus hectori) are a small endangered coastal species that are endemic to New Zealand. Anthropogenic factors, particularly accidental capture in fishing nets, are believed to be the biggest threat to survival of this species. The role of infectious disease as a cause of mortality has not previously been well investigated. This study investigates Toxoplasma gondii infection in Hector's dolphins, finding that 7 of 28 (25%) dolphins examined died due to disseminated toxoplasmosis, including 2 of 3 Maui's dolphins, a critically endangered sub-species. A further 10 dolphins had one or more tissues that were positive for the presence of T. gondii DNA using PCR. Genotyping revealed that 7 of 8 successfully amplified isolates were an atypical Type II genotype. Fatal cases had necrotising and haemorrhagic lesions in the lung (n=7), lymph nodes (n=6), liver (n=4) and adrenals (n=3). Tachyzoites and tissue cysts were present in other organs including the brain (n=5), heart (n=1), stomach (n=1) and uterus (n=1) with minimal associated inflammatory response. One dolphin had a marked suppurative metritis in the presence of numerous intra-epithelial tachyzoites. No dolphins had underlying morbillivirus infection. This study provides the first evidence that infectious agents could be important in the population decline of this species, and highlights the need for further research into the route of entry of T. gondii organisms into the marine environment worldwide.