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1.
World J Gastroenterol ; 22(43): 9525-9533, 2016 Nov 21.
Artículo en Inglés | MEDLINE | ID: mdl-27920473

RESUMEN

AIM: To determine the expression and localization of the electrogenic Na+/HCO3- cotransporter (NBC1) in rat pancreas during development. METHODS: The rat pancreas from postnatal and embryos removed from the uterus of pregnant rats that had been sacrificed by CO2 asphyxiation were used. Rat pancreas from embryonic day (E) 15.5 and E18.5 rat embryos was isolated under a stereomicroscope. Rat pancreas from postnatal (P) days 0, 7, 14, 21 and adult was directly isolated by the unaided eye. The RT-PCR analysis of the NBC1 specific region on rat pancreas tissues from different developmental stages. The two antibodies which target the NBC1 common COOH-terminal region and NH2-terminal region detected a clear band of about 145 kDa in the Western blot analysis. The localization of NBC1 was examined by immuno-fluorescence detection. RESULTS: The results revealed the first peak of NBC1 expression at E18.5 and the second peak at P14. Meanwhile, the low NBC1 expression occurred at P7 and adult stages. Our results demonstrated, for the first time, the presence of NBC1 in the plasma membrane of ß and α cells, as well as in the basolateral membrane of acinar cells of the rat pancreas at different stages of development. CONCLUSION: The data strongly suggests that NBC1 is diversely expressed in the pancreas at different developmental stages, where it may exert its functions in pancreatic development especially islet cell growth through HCO3- transport and pH regulation.


Asunto(s)
Células Secretoras de Glucagón/metabolismo , Células Secretoras de Insulina/metabolismo , Páncreas/metabolismo , Simportadores de Sodio-Bicarbonato/metabolismo , Factores de Edad , Animales , Animales Recién Nacidos , Western Blotting , Técnica del Anticuerpo Fluorescente , Regulación del Desarrollo de la Expresión Génica , Edad Gestacional , Morfogénesis , Páncreas/embriología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas Sprague-Dawley , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Simportadores de Sodio-Bicarbonato/genética
2.
PLoS One ; 11(3): e0149233, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-26974438

RESUMEN

The spread of methicillin-resistant Staphylococcus aureus (MRSA) is a critical health issue that has drawn greater attention to the potential use of immunotherapy. Toll-like receptor 2 (TLR2), a pattern recognition receptor, is an essential component in host innate defense system against S. aureus infection. However, little is known about the innate immune response, specifically TLR2 activation, against MRSA infection. Here, we evaluate the protective effect and the mechanism of MRSA murine pneumonia after pretreatment with Pam3CSK4, a TLR2 agonist. We found that the MRSA-pneumonia mouse model, pretreated with Pam3CSK4, had reduced bacteria and mortality in comparison to control mice. As well, lower protein and mRNA levels of TNF-α, IL-1ß and IL-6 were observed in lungs and bronchus of the Pam3CSK4 pretreatment group. Conversely, expression of anti-inflammatory cytokine IL-10, but not TGF-ß, increased in Pam3CSK4-pretreated mice. Our additional studies showed that CXCL-2 and CXCL1, which are necessary for neutrophil recruitment, were less evident in the Pam3CSK4-pretreated group compared to control group, whereas the expression of Fcγ receptors (FcγⅠ/Ⅲ) and complement receptors (CR1/3) increased in murine lungs. Furthermore, we found that increased survival and improved bacterial clearance were not a result of higher levels of neutrophil infiltration, but rather a result of enhanced phagocytosis and bactericidal activity of neutrophils in vitro and in vivo as well as increased robust oxidative activity and release of lactoferrin. Our cumulative findings suggest that Pam3CSK4 could be a novel immunotherapeutic candidate against MRSA pneumonia.


Asunto(s)
Lipopéptidos/farmacología , Staphylococcus aureus Resistente a Meticilina/inmunología , Neumonía Estafilocócica/tratamiento farmacológico , Receptor Toll-Like 2/agonistas , Animales , Citocinas/inmunología , Antígeno de Macrófago-1/inmunología , Ratones , Neumonía Estafilocócica/inmunología , Neumonía Estafilocócica/patología , Receptores de Complemento 3b/inmunología , Receptores de IgG/inmunología , Receptor Toll-Like 2/inmunología
3.
J Basic Microbiol ; 55(1): 135-40, 2015 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-25590873

RESUMEN

So far, little is known about the diversity of the radiation-resistant microbes of the hyperarid Taklimakan Desert. In this study, ionizing radiation (IR)-resistant bacteria from two sites in Xinjiang were investigated. After exposing the arid (water content of 0.8 ± 0.3%) and non-arid (water content of 21.3 ± 0.9%) sediment samples to IR of 3000 Gy using a (60)Co source, a total of 52 γ-radiation-resistant bacteria were isolated from the desert sample. The 16S rRNA genes of all isolates were sequenced. The phylogenetic tree places these isolates into five groups: Cytophaga-Flavobacterium-Bacteroides, Proteobacteria, Deinococcus-Thermus, Firmicutes, and Actinobacteria. Interestingly, this is the first report of radiation-resistant bacteria belonging to the genera Knoellia, Lysobacter, Nocardioides, Paracoccus, Pontibacter, Rufibacter and Microvirga. The 16s rRNA genes of four isolates showed low sequence similarities to those of the published species. Phenotypic analysis showed that all bacteria in this study are able to produce catalase, suggesting that these bacteria possess reactive oxygen species (ROS)-scavenging enzymes. These radiation-resistant bacteria also displayed diverse metabolic properties. Moreover, their radiation resistances were found to differ. The diversity of the radiation-resistant bacteria in the desert provides further ecological support for the hypothesis that the ionizing-radiation resistance phenotype is a consequence of the evolution of ROS-scavenging systems that protect cells against oxidative damage caused by desiccation.


Asunto(s)
Bacterias/clasificación , Bacterias/efectos de la radiación , Clima Desértico , Rayos gamma , Tolerancia a Radiación , Microbiología del Suelo , Actinobacteria/clasificación , Actinobacteria/genética , Bacterias/genética , Bacterias/aislamiento & purificación , Catalasa/metabolismo , China , Variación Genética , Bacterias Grampositivas/clasificación , Bacterias Grampositivas/genética , Bacterias Grampositivas/aislamiento & purificación , Bacterias Grampositivas/efectos de la radiación , Fenotipo , Filogenia , Proteobacteria/clasificación , Proteobacteria/genética , ARN Ribosómico 16S/genética , Especies Reactivas de Oxígeno/metabolismo
4.
Eukaryot Cell ; 1(2): 191-9, 2002 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-12455954

RESUMEN

Giardia lamblia is medically important as a cause of diarrhea and malabsorption throughout the world and is thought to be one of the earliest-branching eukaryotes on a phylogenetic tree. Nevertheless, the mechanisms of inheritance are largely unknown. The trophozoites of Giardia and other diplomonads are interesting in their possession of two nuclei that are identical or similar in several respects. They replicate at nearly the same time, have similar quantities of DNA, and are both transcriptionally active. We used fluorescence in situ hybridization to demonstrate that genes from each of the five chromosomes are found in both nuclei, confirming that each nucleus has at least one complete copy of the genome. This raises a second question. The alleles of a gene in different nuclei are expected to accumulate different mutations, but surprisingly, the degree of heterozygosity in a clone is very low. One possible mechanism for eliminating sequence differences between nuclei is that each daughter cell receives two copies of the same nucleus at cell division. We used trophozoites with a plasmid transfected into a single nucleus to demonstrate that the two nuclei are partitioned equationally at cytokinesis. The mechanism(s) by which homozygosity is maintained will require further investigation.


Asunto(s)
Núcleo Celular/genética , Genoma de Protozoos , Giardia lamblia/genética , Animales , División Celular/genética , División Celular/fisiología , Núcleo Celular/fisiología , Núcleo Celular/ultraestructura , Sondas de ADN/análisis , Sondas de ADN/genética , ADN Protozoario/análisis , Dosificación de Gen , Giardia lamblia/citología , Giardia lamblia/ultraestructura , Giardiasis/parasitología , Humanos , Microscopía Fluorescente , Hibridación de Ácido Nucleico , Fase S/fisiología , Transfección
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