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2.
Front Immunol ; 13: 1020056, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36569914

RESUMO

Introduction: Intestinal roundworms cause chronic debilitating disease in animals, including humans. Traditional experimental models of these types of infection use a large single-dose infection. However, in natural settings, hosts are exposed to parasites on a regular basis and when mice are exposed to frequent, smaller doses of Heligmosomoides polygyrus, the parasites are cleared more quickly. Whether this more effective host response has any negative consequences for the host is not known. Results: Using a trickle model of infection, we found that worm clearance was associated with known resistance-related host responses: increased granuloma and tuft cell numbers, increased levels of granuloma IgG and decreased intestinal transit time, as well as higher serum IgE levels. However, we found that the improved worm clearance was also associated with an inflammatory phenotype in and around the granuloma, increased smooth muscle hypertrophy/hyperplasia, and elevated levels of Adamts gene expression. Discussion: To our knowledge, we are the first to identify the involvement of this protein family of matrix metalloproteinases (MMPs) in host responses to helminth infections. Our results highlight the delicate balance between parasite clearance and host tissue damage, which both contribute to host pathology. When continually exposed to parasitic worms, improved clearance comes at a cost.


Assuntos
Nematospiroides dubius , Humanos , Camundongos , Animais , Cicatriz , Imunidade , Granuloma , Inflamação
3.
J Leukoc Biol ; 108(1): 83-91, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32170880

RESUMO

Eosinophils are traditionally associated with allergic and parasitic inflammation. More recently, eosinophils have also been shown to have roles in diverse processes including development, intestinal health, thymic selection, and B-cell survival with the majority of these insights being derived from murine models and in vitro assays. Despite this, tools to measure the dynamic activity of eosinophils in situ have been lacking. Intravital microscopy is a powerful tool that enables direct visualization of leukocytes and their dynamic behavior in real-time in a wide range of processes in both health and disease. Until recently eosinophil researchers have not been able to take full advantage of this technology due to a lack of tools such as genetically encoded reporter mice. This mini-review examines the history of intravital microscopy with a focus on eosinophils. The development and use of eosinophil-specific Cre (EoCre) mice to create GFP and tdTomato fluorescent reporter animals is also described. Genetically encoded eosinophil reporter mice combined with intravital microscopy provide a powerful tool to add to the toolbox of technologies that will help us unravel the mysteries still surrounding this cell.


Assuntos
Eosinófilos/citologia , Microscopia Intravital , Animais , Ceco/citologia , Corantes Fluorescentes/metabolismo , Genes Reporter , Intestino Delgado/citologia , Pulmão/citologia , Linfonodos/citologia , Camundongos Endogâmicos C57BL , Músculos/citologia
4.
Clin Chim Acta ; 478: 37-43, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29258744

RESUMO

BACKGROUND: In vitro deamination generates ammonia in freshly collected blood specimens. To prevent this, samples for ammonia testing are usually collected on ice and run rapidly (e.g., within 1h). We developed a method to stabilize specimens for ammonia analysis. METHODS: Following plasma separation, 500µmol/l cycloserine or a combination of 2mmol/l sodium borate with 5mmol/l l-serine were added to sample pools with normal or increased concentrations of ALT and/or GGT to inhibit deamination; and/or residual platelets were removed via centrifugation. Sample pools were then incubated at room temperature or 4°C. Untreated sample pools were also incubated at -80°C. Ammonia was measured at 0, 1, 2, 4, 8, 16, and 24h. RESULTS: When incubated at 4°C without treatment, sample pools with enzymes within their reference limit had an increase of 0.5µmol/l/h, whereas sample pools with ALT and/or GGT activity above their upper reference limit had an increase of 3.6µmol/l/h (p<0.001). When sample pools were incubated at 4°C with sodium borate/l-serine, the rate of ammonia increase was significantly reduced in samples with normal (0.3µmol/l/h, p<0.001 vs. untreated controls) or high enzyme activity (0.1µmol/l/h, p<0.001 vs. untreated controls). Independent of the ALT and/or GGT concentrations, storing the sample at -80°C also preserved the specimens for ammonia analysis (0.2µmol/l/h, p<0.001 vs. untreated controls). CONCLUSIONS: By combining sodium borate/l-serine with refrigeration, plasma ammonia specimens can be stabilized for >12h.


Assuntos
Amônia/sangue , Manejo de Espécimes/métodos , Amônia/química , Coleta de Amostras Sanguíneas/métodos , Boratos , Serviços de Laboratório Clínico , Criopreservação/métodos , Humanos , Serina , Temperatura
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