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1.

Background aims

Multipotent mesenchymal stromal cell (MSC)-based medicines are extensively investigated for use in regenerative medicine and immunotherapy applications. The International Society for Cell and Gene Therapy (ISCT) proposed a panel of cell surface molecules for MSC identification that includes human leukocyte antigen (HLA)-DR as a negative marker. However, its expression is largely unpredictable despite production under tightly controlled conditions and compliance with current Good Manufacturing Practices. Herein, we report the frequency of HLA-DR expression in 81 batches of clinical grade bone marrow (BM)-derived MSCs and investigated its impact on cell attributes and culture environment.

Methods

The levels of 15 cytokines (interleukin [IL]-1β, IL-4, IL-6, IL-10, IL-17A, IL-17F, IL-21, IL-22, IL-23, IL-25, IL-31, IL-33, interferon-γ, soluble CD40 ligand and tumor necrosis factor-α) were determined in sera supplements and supernatants of BM-MSC cultures. Identity, multipotentiality and immunopotency assays were performed on high (>20% of cells) and low (≤20% of cells) HLA-DR+ cultures.

Results

A correlation was found between HLA-DR expression and levels of IL-17F and IL-33. Expression of HLA-DR did neither affect MSC identity, in vitro tri-lineage differentiation potential (into osteogenic, chondrogenic and adipogenic lineages), nor their ability to inhibit the proliferation of stimulated lymphocytes.

Discussion

Out of 81 batches of BM-MSCs for autologous use analyzed, only three batches would have passed the ISCT criteria (<2%), whereas 60.5% of batches were compliant with low HLA-DR values (≤20%). Although a cause–effect relationship cannot be drawn, we have provided a better understanding of signaling events and cellular responses in expansion culture conditions relating with HLA-DR expression.  相似文献   
2.
Three-day-old cowpea seedlings were inoculated with the severestrain of Cowpea Mosaic Virus (CpMV) and 24 h later with Bradyrhizobiumsp. cowpea, strain I-125, when virus translocation to rootsstill had not taken place. Plants were harvested at 22, 30,45 and 59 d after germination. Active virus replication wasassociated with increased protein content, detected in the leavesof 22-d-old plants. CpMV infection reduced the total leaf area,dry weight of shoots and the chlorophyll content of the firsttrifoliolate leaf at all experimental times. Low sugar contentwas recorded in leaves of 22- and 30-d-old CpMV-infected plantsand in nodules and roots of 30- and 45-d-old CpMV-infected plants.Up to 45 d, the nodule mass of CpMV-infected plants was lowerthan in controls, but reached control values at the 4th harvest.In CpMV-infected nodules, massive agglomeration of virus particles,crystalline virus inclusions and the proliferation of the endoplasmicreticulum were observed only in those cells containing bacteroids.In 30- and 45-d-old plants, CpMV infection decreased the contentof ureides in nodules, roots, and petioles. Virus infectiondid not alter the -amino-N content of roots and nodules butinduced a transient 74% reduction in the level of -amino-N inpetioles of 45-d-old plants. At the 1st and 2nd harvests theactivity of uricase (EC 1.7.3.3 [EC] ) in the nodules and of pyruvatekinase (EC 2.7.1.40 [EC] ) in the nodules and leaves were decreasedseverely by virus infection. CpMV did not hinder the allantoinase(EC 3.5.2.5 [EC] ) activity in the leaves but caused a 9% transitorydecrease in the activity of this enzyme in nodules of 45-d-oldplants. Measurements of NAD-malic enzyme (EC 1.1.1.38 [EC] ) in nodulesalso showed the non-effect of CpMV on this enzyme, except fora temporary 16% reduction at the 2nd harvest. As compared tocontrols, the relative abundance of ureides in 30-d-old CpMV-infectedplants indicated a 15%, 10%, and 51% reduction in the nodules,roots, and petioles, respectively. Results indicate that atthe time of the 4th harvest the symbiotic process, measuredin terms of ureide content and enzymatic activities, was functioningat a near normal level despite nodule infection by CpMV. Key words: Cowpea Mosaic Virus, nitrogen fixation, cowpea, enzymes, ultrastructure  相似文献   
3.
Abstract In this study, we show that the house mouse (Mus musculus) is the most abundant small mammal in the National Reserve of Lachay in central Peru, and that its large population fluctuations are independent of seasonality. Also, we found that M. musculus is the main small mammal prey of burrowing owls (Athene cunicularia) in Lachay, and that owls respond functionally to mouse abundance. In addition, vegetation cover seems to have a strong effect on small mammal predation by burrowing owls, and possibly other predators such as eagles and foxes. We propose the hypothesis that burrowing owls in arid environments can survive several months eating only arthropods, but that their reproduction is synchronized with a highly nutritious diet, provided by small mammal consumption. The principal prediction of this alternative prey hypothesis is now supported by our data from a tropical ecosystem.  相似文献   
4.
Coconut (Cocos nucifera L.) WRKY sequences containing single nucleotide polymorphisms (SNPs) and one microsatellite repeat were used to develop 10 informative markers. These markers were evaluated in 15 genotypes representing six coconut cultivars. SNP‐containing alleles were detected by single‐strand conformation polymorphism (SSCP) analysis. The number of detected alleles ranged from two to four. Five pairs of loci were in linkage disequilibrium in the test population. These markers are currently being evaluated in more individuals/cultivars to determine their value in estimating the genetic diversity of this species.  相似文献   
5.
Phylogenetic information is useful in understanding the evolutionary history of adaptive traits. Here, we present a well-resolved phylogenetic hypothesis for Heliconius butterflies and related genera. We use this tree to investigate the evolution of three traits, pollen feeding, pupal-mating behaviour and larval gregariousness. Phylogenetic relationships among 60 Heliconiina species (86% of the subtribe) were inferred from partial DNA sequences of the mitochondrial genes cytochrome oxidase I , cytochrome oxidase II and 16S rRNA, and fragments of the nuclear genes elongation factor-1α , apterous , decapentaplegic and wingless (3834 bp in total). The results corroborate previous hypotheses based on sequence data in showing that Heliconius is paraphyletic, with Laparus doris and Neruda falling within the genus, demonstrating a single origin for pollen feeding but with a loss of the trait in Neruda . However, different genes are not congruent in their placement of Neruda ; therefore, monophyly of the pollen feeding species cannot be ruled out. There is also a highly supported monophyletic 'pupal-mating clade' suggesting that pupal mating behaviour evolved only once in the Heliconiina. Additionally, we observed at least three independent origins for larval gregariousness from a solitary ancestor, showing that gregarious larval behaviour arose after warning coloration.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 92 , 221–239.  相似文献   
6.
Comparisons between historical and recent ecological datasets indicate that shallow reef habitats across the central Galapagos Archipelago underwent major transformation at the time of the severe 1982/1983 El Niño warming event. Heavily grazed reefs with crustose coralline algae (‘urchin barrens’) replaced former macroalgal and coral habitats, resulting in large local and regional declines in biodiversity. Following recent threat assessment workshops, a total of five mammals, six birds, five reptiles, six fishes, one echinoderm, seven corals, six brown algae and nine red algae reported from coastal environments in Galapagos are now recognized as globally threatened. The 2008 International Union for the Conservation of Nature (IUCN) Red List includes 43 of these species, while two additional species (Galapagos damsel Azurina eupalama and 24‐rayed sunstar Heliaster solaris) not seen for > 25 years also fulfil IUCN threatened species criteria. Two endemic species (Galapagos stringweed Bifurcaria galapagensis and the damselfish A. eupalama) are now regarded as probably extinct, while an additional six macroalgal species (Dictyota galapagensis, Spatoglossum schmittii, Desmarestia tropica, Phycodrina elegans, Gracilaria skottsbergii and Galaxaura barbata) and the seastar H. solaris are possibly extinct. The removal of large lobster and fish predators by artisanal fishing probably magnified impacts of the 1982/1983 El Niño through a cascade of indirect effects involving population expansion of grazing sea urchins. Marine protected areas with adequate enforcement are predicted to ameliorate but not eliminate ecosystem impacts caused by increasing thermal anomalies associated with El Niño and global climate change.  相似文献   
7.
Abstract The knowledge of dispersal is essential to understand the ecology of any species, since population dynamics, spatial distribution and genetic structure are closely tied to patterns of movement. In this paper we estimate dispersal patterns in natural populations of the endangered snake Boa constrictor occidentalis (Serpentes, Boidae), using allozymes as genetic markers. Blood samples were obtained from a total of 120 adult individuals of nine localities from two areas, Sobremonte and Pocho, in Argentina. Only four out of a total of 24 loci were polymorphic: 6‐Pgdh‐1, Cat‐1, Ldh and Hp. The values of expected and observed mean heterozygosities, the percentage of polymorphic loci and mean number of alleles per locus for each population confirm the low levels of genetic variability in this snake. FST and Nem mean values were 0.0089 and 46.1 for Sobremonte and 0.0379 and 7.46 for Pocho, indicating important levels of gene flow. A comparison of FST between genders in both areas suggests a male‐biased dispersal, which could be explained by the characteristics of the mating system: males carry out an active mate search of receptive females, and philopatry could be selected in females due to the benefits of the familiarity with the natal area in the use of local resources.  相似文献   
8.
The ontogeny of brain creatine kinase (CK) was studied during chick embryo development. The cytosolic activity increased 270% in 10 h from the 2nd to the 3rd days of incubation; this was followed by a plateau phase throughout development and at the end of incubation there appeared to be another increase of cytosolic and mitochondrial CK activities. Therefore, early embryonic chick brain CK is another‘constitutive’enzyme like the early embryonic chick heart CK since creatine has not been enzymatically detected in the embryo until day 4 of incubation. Insulin does not appear to stimulate the early increase of brain CK activity since the hormone is not present in the embryo until day 5 of incubation. It is likely that CK increase is associated with neuronal multiplication at early stages and possibly to neuronal maturation before hatching.  相似文献   
9.
The effect of infection by the Cowpea Mosaic Virus (CpMV) onseveral parameters relevant to symbiotic nitrogen fixation wasdetermined in cowpea (Vigna unguiculata (L.) Walp. var. Tuy)plants nodulated with two strains of Rhizobium cowpea: IVIC–124and IVIC–38. Plants were virus-infected at the seedlingstage before Rhizobium inoculation. The effect of CpMV infectionon plant growth was analysed in nodulated and nitrogen-suppliedplants at 18, 25 and 35 d after germination. At all developmentalstages of nodulated plants CpMV infection caused a reductionof leaf chlorophyll content, leaf area, dry weight of shootsand roots, total nodule weight and nodule number. Most of thenodules from 18- and 25-d-old CpMV-infected plants did not exhibitleghaemoglobin pigmentation. CpMV infection delayed the onsetof nitrogenase activity in nodules of the rhizobial strain IVIC–124and the enzyme activity measured on a per plant basis was reducedin both strains at the first and second harvests. Significantnitrogenase activity was detected in 35-d-old infected plants.Some of the nodules of the rhizobial strain IVIC-124 and mostof the nodules from plants nodulated with the strain IVIC-38developed leghaemoglobin; however, the nodule-specific nitrogenaseactivity, estimated on a milligram nodule dry weight basis,was always higher in virus-infected plants, particularly in18-d-old CpMV-infected plants harbouring the IVIC–124strain. CpMV-infected nodules had a larger peribacteroidal space,a reduced number of peribacteroid units, a greater number ofbacteroids per unit, a lower number of vesicles and 88% lowertotal reducing sugar content. Starch accumulation was detectedin infected leaves of nodulated plants during the first harvest,while high levels of leaf reducing sugars and protein were presentat the second harvest. In healthy nodulated plants the rhizobialstrain IVIC–124 was shown to be more efficient than IVIC–38in promoting plant growth. However, the results indicate thatnodulation by rhizobial strain IVIC–124 and growth ofplants harbouring this strain were affected to a greater extentby virus infection. The effect of CpMV infection on leaf chlorophyllcontent, leaf area, carbohydrate level, leaf proteins and growthof nitrogen-supplied plants, as well as the symptoms inducedin the leaves, were less conspicuous than in nodulated plants. Key words: Cowpea, Rhizobium, virus infection, nodule untrastructure  相似文献   
10.
We measured available and actual habitat use, morphology, escape behaviour and clinging ability in a large sample ( N  = 242) of green anoles, Anolis carolinensis , in a habitat consisting primarily of segregated dense clumps of broad leaves, Aspidistra elatior (Tulane University campus, LA) to compare against similar data collected previously from a more typical habitat c. 30 km away, consisting of continuous strands of bushes and trees (Good Hope Field, St. Charles Parish, LA). At Tulane the anoles perched primarily on the broad, smooth leaves of broad leaves, whereas in Good Hope Field (GHF) they predominantly perched on branches and tree trunks. The two populations differed significantly in morphology. In Tulane, the anoles tended to have shorter distal hindlimb elements, longer forelimb elements, and were more 'slender' than those at GHF. A comparison of escape behaviour showed population and sex differences. In both populations, females had significantly longer approach distances (i.e. were more 'wary') than males. These distances were, in addition, significantly longer at GHF than at Tulane for both sexes; this may be due to the potentially higher diversity and abundance of predators at GHF, although habituation to humans may also play a role. Anoles at Tulane had significantly larger toepads and higher clinging abilities than those at GHF. The enhanced clinging abilities of anoles at Tulane may have arisen due to their propensity to use smooth leaves as their primary substrate. Overall, our data reveal substantial ecological, behavioural, morphological, and functional differences among populations, some of which may be adaptive.  © 2005 The Linnean Society of London, Biological Journal of the Linnean Society , 2005, 85 , 223–234.  相似文献   
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