We investigated the in
vitro differentiation of canine bone marrow stromal cells (BMSCs) into voltage-
and glutamate-responsive neuron-like cells. BMSCs were obtained from the bone marrow of
healthy beagle dogs. Canine BMSCs were incubated with the basal medium for neurons
containing recombinant human basic fibroblast growth factor (bFGF; 100
ng/ml). The viability of the bFGF-treated cells was
assessed by a trypan blue exclusion assay, and the morphology was monitored. Real-time
RT-PCR was performed to evaluate mRNA expression of neuronal, neural stem cell and glial
markers. Western blotting and immunocytochemical analysis for the neuronal markers were
performed to evaluate the protein expression and localization. The Ca2+
mobilization of the cells was evaluated using the Ca2+ indicator Fluo3 to
monitor Ca2+ influx. To investigate the mechanism of bFGF-induced neuronal
differentiation, the fibroblast growth factor receptor inhibitor, the phosphoinositide
3-kinase inhibitor or the Akt inhibitor was tested. The bFGF treatment resulted in the
maintenance of the viability of canine BMSCs for 10 days, in the expression of neuronal
marker mRNAs and proteins and in the manifestation of neuron-like morphology. Furthermore,
in the bFGF-treated BMSCs, a high concentration of KCl and L-glutamate induced an increase
in intracellular Ca2+ levels. Each inhibitor significantly attenuated the
bFGF-induced increase in neuronal marker mRNA expression. These results suggest that bFGF
contributes to the differentiation of canine BMSCs into voltage- and glutamate-responsive
neuron-like cells and may lead to the development of new cell-based treatments for
neuronal diseases. 相似文献
LC3 − the mammalian homolog of Atg8 − was found as autophagosome membrane binding protein
in mammals and widely used as an autophagosomal marker. LC3A, B and C show different
expression patterns in each tissue. The aim of this study was to reveal the differences of
expression patterns among LC3 families in mouse placenta under normal condition and
nutrient starving condition. LC3A and B were highly expressed in decidual cells. LC3A and
B were increased in D14 compared with D12 and D16 in mouse placenta, while LC3C was
decreased. Starvation induced increase in LC3B expression specifically.
Immunohistochemistry showed different expression patterns among LC3A, B and C. LC3A
expression in syncytiotrophoblast was vanished by starvation. The results of real time
RT-PCR suggested differences between D12 and D16 in autophagic cascade induced by
starvation. Taken together, this study suggests that autophagy could play a role in
placental invasion system and that nutrient starvation affects LC3B expression. 相似文献
L-type amino acid transporter
1 (LAT1), the first isotype of amino acid transport system L, transports aromatic and
branched amino acids pivotal for fundamental cellular activities such cellular growth and
proliferation. LAT1 expression was high only in the brain in contrast to its limited
distribution and low level of expression in normal tissues. We found potent LAT1
expression in canine caput epididymis by quantitative RT-PCR and Western blotting
analysis. Immnuno-histochemical examination revealed observable LAT1 in microvillous
epithelial cells. 相似文献
The ecological literature has documented the effects of plant hybridization on phenotypic variation, and dominant, intermediate, or novel morphological, chemical and physiological traits in hybrids. It is important to understand the ecological consequences of hybridization by evaluating their impact on phenotypic expression of functional traits. We evaluated the relationship between genetic diversity of Quercus laurina and functional foliar traits along an oak diversity gradient. We selected five study sites that represent an oak diversity gradient where Q. laurina is present. Using chloroplast and nuclear microsatellites, we evaluated genetic diversity, measured functional foliar traits of Q. laurina in each site and assessed the effects of local climate variables on the oak community and functional traits. We found a greater abundance of Q. laurina in all study sites. We did not find a relationship between the number of accompanying red oak species and the population genetic diversity in Q. laurina, but higher genetic diversity was found in all study sites in comparison with European oak species. Sites with more oak species had more variation of foliar functional traits. Our results do not support the hypothesis that predicts higher levels of genetic diversity of Q. laurina in communities with greater oak diversity from the same section, but we demonstrated an increase in the foliar functional traits of Q. laurina associated with oak richness and climate variables. We highlight the need to consider environmental and ecological variables linkages as regulatory mechanisms of the phenotypic plasticity expressed in changes of some functional attributes of oaks.
Post-operative pain management following rumen surgery is not common practice. We examined the effect of providing the pain medication ketoprofen to dairy cattle following the first stage of a rumen cannulation surgery, which involves an incision in the body wall and exteriorizing and clamping the rumen. The results of this study provide clear evidence that the first stage of the surgery was painful and ketoprofen at the time of and 24 h following surgery, alleviated some, but not all, of the post-surgical pain. Pain mitigation should be included when performing flank surgery in cattle. 相似文献
The major indication for antibiotic use in Danish pigs is treatment of intestinal diseases post weaning. Clinical decisions on antibiotic batch medication are often based on inspection of diarrhoeic pools on the pen floor. In some of these treated diarrhoea outbreaks, intestinal pathogens can only be demonstrated in a small number of pigs within the treated group (low pathogen diarrhoea). Termination of antibiotic batch medication in herds suffering from such diarrhoea could potentially reduce the consumption of antibiotics in the pig industry. The objective of the present pilot study was to suggest criteria for herd diagnosis of low pathogen diarrhoea in growing pigs.Data previously collected from 20 Danish herds were used to create a case series of clinical diarrhoea outbreaks normally subjected to antibiotic treatment. In the present study, these diarrhoea outbreaks were classified as low pathogen (<15% of the pigs having bacterial intestinal disease) (n =5 outbreaks) or high pathogen (≥15% of the pigs having bacterial intestinal disease) (n =15 outbreaks). Based on the case series, different diagnostic procedures were explored, and criteria for herd diagnosis of low pathogen diarrhoea were suggested. The effect of sampling variation was explored by simulation.
Results
The diagnostic procedure with the highest combined herd-level sensitivity and specificity was qPCR testing of a pooled sample containing 20 randomly selected faecal samples. The criteria for a positive test result (high pathogen diarrhoea outbreak) were an average of 1.5 diarrhoeic faecal pools on the floor of each pen in the room under investigation and a pathogenic bacterial load ≥35,000 per gram in the faecal pool tested by qPCR. The bacterial load was the sum of Lawsonia intracellularis, Brachyspira pilosicoli and Escherichia coli F4 and F18 bacteria per gram faeces. The herd-diagnostic performance was (herd-level) diagnostic sensitivity =0.99, diagnostic specificity =0.80, positive predictive value =0.94 and negative predictive value =0.96.
Conclusions
The pilot study suggests criteria for herd diagnosis of low pathogen diarrhoea in growing pigs. The suggested criteria should now be evaluated, and the effect of terminating antibiotic batch medication in herds identified as suffering from low pathogen diarrhoea should be explored.
Electronic supplementary material
The online version of this article (doi:10.1186/2046-0481-67-24) contains supplementary material, which is available to authorized users. 相似文献