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41.
Cells act as extremely efficient filters for elution of unbound fluorescent tags or impurities associated with nanoparticles, including those that cannot be removed by extensive cleaning. This has consequences for quantification of nanoparticle uptake and sub-cellular localization in vitro and in vivo as a result of the presence of significant amount of labile dye even following extensive cleaning by dialysis. Polyacrylamide gel electrophoresis (PAGE) can be used to monitor the elution of unbound fluorescent probes from nanoparticles, either commercially available or synthesized in-house, and to ensure their complete purification for biological studies, including cellular uptake and sub-cellular localisation. Very different fluorescence distribution within cells is observed after short dialysis times versus following extensive dialysis against a solvent in which the free dye is more soluble, due to the contribution from free dye. In the absence of an understanding of the presence of residual free dye in (most) labeled nanoparticle solutions, the total fluorescence intensity in cells following exposure to nanoparticle solutions could be mis-ascribed to the presence of nanoparticles through the cell, rather than correctly assigned to either a combination of free-dye and nanoparticle-bound dye, or even entirely to free dye depending on the exposure conditions (i.e. aggregation of the particles etc). Where all of the dye is nanoparticle-bound, the particles are highly localized in sub-cellular organelles, likely lysosomes, whereas in a system containing significant amounts of free dye, the fluorescence is distributed through the cell due to the free diffusion of the molecule dye across all cellular barriers and into the cytoplasm. 相似文献
42.
M G Narducci D Arcelli M C Picchio C Lazzeri E Pagani F Sampogna E Scala P Fadda C Cristofoletti A Facchiano M Frontani A Monopoli M Ferracin M Negrini G A Lombardo E Caprini G Russo 《Cell death & disease》2011,2(4):e151
Sézary syndrome (SS) is an incurable leukemic variant of cutaneous T-cell lymphoma and its pathogenesis is still unknown. Diagnosis/prognosis may strongly ameliorate the management of SS individuals. Here, we profiled the expression of 470 microRNAs (miRNAs) in a cohort of 22 SS patients, and we identified 45 miRNAs differentially expressed between SS and controls. Using predictive analysis, a list of 19 miRNAs, including miR-21, miR-214, miR-486, miR-18a, miR-342, miR-31 and let-7 members were also found. Moreover, we defined a signature of 14 miRNAs including again miR-21, potentially able to discriminate patients with unfavorable and favorable outcome. We validated our data for miR-21, miR-214 and miR-486 by qRT-PCR, including an additional set of array-independent SS cases. In addition, we also provide an in vitro evidence for a contribution of miR-214, miR-486 and miR-21 to apoptotic resistance of CTCL cell line. 相似文献
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44.
MP Lisovoi NM Lesovoy GI Vasechko 《Archives Of Phytopathology And Plant Protection》2013,46(2):123-127
A new method of selection of the winter wheat varieties has been tested for resistance to the pest insects' complex by the traits of plants that are the markers of plant resistance. It makes it possible to use this method from year to year independently of the pests' density. 相似文献
45.
TD Smith KP Bhatnagar CJ Bonar KL Shimp MP Mooney MI Siegel 《American journal of physical anthropology》2003,122(3):301-301
46.
The aim of this experiment was to evaluate the effects of an intracerebroventricular (icv) injection of prostaglandin E1 (PGE1) on the sympathetic activation and the thermogenic changes in rats with acute lesions of the ventromedial hypothalamus (VMH). Four groups of six Sprague-Dawley male rats were anesthetized with ethyl-urethane. The firing rate of the sympathetic nerves innervating the interscapular brown adipose tissue (IBAT) and the colonic and IBAT temperatures were monitored both before and after one of the following treatments: 1) VMH lesion plus icv injection of PGE1 (500 ng); 2) VMH lesion plus icv injection of saline; 3) sham lesion plus icv injection of PGE1; and 4) sham lesion plus icv injection of saline. PGE1 induced an increase in the firing rate of IBAT nerves and the colonic and IBAT temperatures. These effects were reduced by VMH lesion. The findings indicate that acute lesions of the VMH reduce the effects of PGE1 and seem to suggest a possible role played by the VMH in the control of the sympathetic activation and the thermogenic changes during PGE1 hyperthermia. 相似文献
47.
Multiple haplotypes from each of three nuclear loci were isolated and
sequenced from geographic populations of the American oyster, Crassostrea
virginica. In tests of alternative phylogeographic hypotheses for this
species, nuclear gene genealogies constructed for these haplotypes were
compared to one another, to a mitochondrial gene tree, and to patterns of
allele frequency variation in nuclear restriction site polymorphisms
(RFLPs) and allozymes. Oyster populations from the Atlantic versus the Gulf
of Mexico are not reciprocally monophyletic in any of the nuclear gene
trees, despite considerable genetic variation and despite large allele
frequency differences previously reported in several other genetic assays.
If these populations were separated vicariantly in the past, either
insufficient time has elapsed for neutral lineage sorting to have achieved
monophyly at most nuclear loci, or balancing selection may have inhibited
lineage extinction, or secondary gene flow may have moved haplotypes
between regions. These and other possibilities are examined in light of
available genetic evidence, and it is concluded that no simple explanation
can account for the great variety of population genetic patterns across
loci displayed by American oysters. Regardless of the source of this
heterogeneity, this study provides an empirical demonstration that
different sequences of DNA within the same organismal pedigree can have
quite different phylogeographic histories.
相似文献
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49.
Crustacean and cheliceratan hemocyanins (oxygen-transport proteins) and
insect hexamerins (storage proteins) are homologous gene products, although
the latter do not bind oxygen and do not possess the copper- binding
histidines present in the hemocyanins. An alignment of 19 amino acid
sequences of hemocyanin subunits and insect hexamerins was made, based on
the conservation of elements of secondary structure observed in X-ray
structures of two hemocyanin subunits. The alignment was analyzed using
parsimony and neighbor-joining methods. Results provide strong indications
for grouping together the sequences of the 2 crustacean hemocyanin
subunits, the 5 cheliceratan hemocyanin subunits, and the 12 insect
hexamerins. Within the insect clade, four methionine- rich proteins, four
arylphorins, and two juvenile hormone-suppressible proteins from
Lepidoptera, as well as two dipteran proteins, form four separate groups.
In the absence of an outgroup sequence, it is not possible to present
information about the ancestral state from which these proteins are
derived. Although this family of proteins clearly consists of homologous
gene products, there remain striking differences in gene organization and
site of biosynthesis of the proteins within the cell. Because studies on
18S and 12S rRNA sequences indicate a rather close relationship between
insects and crustaceans, we propose that hemocyanin is the ancestral
arthropod protein and that insect hexamerins lost their copper-binding
capability after divergence of the insects from the crustaceans.
相似文献
50.
This paper looked at the leaf architecture characteristics of Vachellia tortilis to determine if either there is or not an effect of the tropic line on plants. Vachellia tortilis leaves were sampled along a national road (N1) in Limpopo province. Sampling points were set 10 km apart away from the Tropic of Capricon in opposite directions. Leaf morphology revealed that leaves of V. tortilis are bipinnately compound with alternate arrangement. The venation pattern of the pinnules was eucamptodromus and brochidodromous with imperfect reticulation. Areoles were imperfect and pentagonal or irregular in shape. 相似文献