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141.
Biotinyl-oligosaccharides are a relatively new generation of saccharide
probes that enable immobilization of desired oligosaccharides on
streptavidin matrices for studies of carbohydrate-protein interactions.
Here we describe the facile preparation of biotinyl-l-3-(2-naphthyl)-
alanine hydrazide (BNAH) derivatives of oligosaccharides, containing a
strong UV absorbing and fluorescent group, in which the ring of the
reducing-end monosaccharide is nonreduced. We evaluate reactivities of
immobilized BNAH- N -glycans with plant lectins that recognize aspects of
the oligosaccharide core or outer-arms. We make some comparisons with
2-amino-6-amidobiotinyl-pyridine (BAP) derivatives obtained by reductive
amination, and 6-(biotinyl)-aminocaproyl-hydrazide (BACH) derivatives which
have a longer spacer-arm. N -Glycan-BNAH and-BAP derivatives have, overall,
comparable reactivities with lectins which recognize N -glycan outer-arms
or the trimannosyl core, but only BNAH and BACH derivatives are bound by
lectins which recognize the non- reduced core. Moreover, with Pisum sativum
agglutinin (PSA) which additionally requires the fucosyl- N-
glycan-asparaginyl core for high affinity binding, the immobilized BNAH
derivative (which is an alanine hydrazide beta-glycoside) can substitute
for the natural beta- glycosylasparaginyl core, whereas the BACH derivative
(aminocaproyl- hydrazide-beta-glycoside) is less effective. BNAH is a
derivatization reagent of choice, therefore, for solid phase
carbohydrate-binding experiments with immobilized N -glycans.
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142.
Monophyly of the order Rodentia inferred from mitochondrial DNA sequences of the genes for 12S rRNA, 16S rRNA, and tRNA-valine 总被引:3,自引:2,他引:1
A recent analysis of amino acid sequence data (Graur et al.) suggested that
the mammalian order Rodentia is polyphyletic, in contrast to most
morphological data, which support rodent monophyly. At issue is whether the
hystricognath rodents, such as the guinea pig, represent an independent
evolutionary lineage within mammals, separate from the sciurognath rodents.
To resolve this problem, we sequenced a region (2,645 bp) of the
mitochondrial genome of the guinea pig containing the complete 12S
ribosomal RNA, 16S ribosomal RNA, and transfer RNA(VAL) genes for
comparison with the available sciurognath and other mammalian sequences.
Several methods of analysis and statistical tests of the data all show
strong support for rodent monophyly (91%-98% bootstrap probability, or BP).
Calibration with the mammalian fossil record suggests a Cretaceous date
(107 mya) for the divergence of sciurognaths and hystricognaths. An older
date (38 mya) for the controversial Mus- Rattus divergence also is
supported by these data. Our neighbor-joining analyses of all available
sequence data (25 genes) confirm that some individual genes support rodent
polyphyly but that tandem analysis of all data does not. We propose that
the conflicting results are due to several compounding factors. The unique
biochemical properties of some hystricognath metabolic proteins, largely
responsible for generating this controversy, may have a single explanation:
a cascade effect resulting from inactivation of the zinc-binding abilities
of insulin. After excluding six genes possibly affected by insulin
inactivation, analyses of all available sequence data (7,117 nucleotide
sites, 3,099 amino acid sites) resulted in strong support for rodent
monophyly (94% BP for DNA sequences, 90% for protein sequences), which
lends support to the insulin-cascade hypothesis.
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143.
144.
145.
The construction of cell hairs (trichomes) on the wings of Drosophila occurs in synchrony on 30,000 cells over a period of about 20 hr. Changes in both morphology and patterns of protein synthesis occur rapidly during this time period. In this report we describe the use of stress-induced (heat shock) abnormalities in morphogenesis to provide further details on the stepwise processes of differentiation within single wing cells. A cartoon summary of the overall process and a discussion of some possible mechanisms is included. 相似文献
146.
Ganoderma microsporum immunomodulatory protein induces apoptosis and potentiates mitomycin C‐induced apoptosis in urinary bladder urothelial carcinoma cells
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147.
Rossi MS; Barrio E; Latorre A; Quezada-Diaz JE; Hasson E; Moya A; Fontdevila A 《Molecular biology and evolution》1996,13(2):314-323
Both original and colonizer populations of Drosophila buzzatii have been
analyzed for mtDNA restriction polymorphisms. Most of the mtDNA nucleotide
variation in original populations of NW Argentina can be explained by
intrapopulation diversity and only a small fraction can be accounted for by
between-population diversity. Similar results are obtained using either the
estimated number of nucleotide substitutions per site or considering each
restriction site as a locus. Colonizer populations of the Iberian Peninsula
are monomorphic and show only the most common haplotype from the original
populations. Under the infinite island model and assuming that populations
are in equilibrium, fixation indices indicate enough gene flow to explain
why the populations are not structured. Yet, the possibility exists that
populations have not reached an equilibrium after a founder event at the
end of the last Pleistocene glaciation. Tajima's test suggests that
directional selection and/or a recent bottleneck could explain the present
mtDNA differentiation. Considering the significant population structure
found for the chromosomal and some allozyme polymorphisms, the among-
population uniformity for mtDNA variability argues in favor of the
chromosomal and some allozyme polymorphisms being adaptive.
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148.
Adipocyte lipolysis is dependent on an increase in the intracellular concentration of cAMP. Intracellular phosphodiesterases (PDEs) hydrolyze cAMP and limit stimulation of lipolysis. In the present study, the mRNA expression of PDE4 subtypes and the antilipolytic role of PDE4 in rat adipocytes were investigated. Fragments encoding PDE4A (233 bp), PDE4B (786 bp), PDE4C (539 bp), and PDE4D (262 bp) sequences were amplified by RT-PCR. The mRNA expression of PDE4 subtypes (A, B, C, D) determined by real-time quantitative PCR was 7, 18.7, 18.9, and 7.2% relative to PDE3B. Inhibition of PDE4 by rolipram increased basal lipolysis and reversed in part prostaglandin E2 antilipolysis. The combination of PDE3 and PDE4 inhibitors synergistically reversed both prostaglandin E2 and phenylisopropyl adenosine antilipolysis. Stimulation of adipocytes with prostaglandin E2 increased total PDE activity and PDE3 activity measured by hydrolysis of 3[H]cAMP by the particulate fraction of adipocytes. The present study confirmed that mRNAs for all four PDE4 subtypes were expressed in rat adipocytes, with PDE4B and PDE4C predominant. Moreover, PDE4 not only limits the rate of basal lipolysis but also contributes to prostaglandin E2 antilipolysis in rat adipocytes. 相似文献
149.
Amrita Ahluwalia PhD Michael K. Jones PhD Neil Hoa MS Andrzej S. Tarnawski MD PhD 《Journal of cellular biochemistry》2019,120(7):11651-11659
Gastric epithelial cells are important components of mucosal protection and targets of nonsteroidal anti-inflammatory drugs (NSAIDs)-induced injury. Diclofenac (DFN) is one of the most widely used NSAIDs; however, even its short-term use can induce gastric erosions and ulcers. Nerve growth factor (NGF) has been reported to act not only on neuronal cells but also on endothelial cells; however, its action on gastric epithelial cells is unknown. This study was aimed to determine, whether NGF can protect gastric epithelial cells against DFN-induced injury, and to determine the underlying molecular mechanisms with a focus on mitochondria, survivin, and insulin-like growth factor 1 (IGF-1). Cultured normal rat gastric mucosal epithelial cells 1 (RGM1) were treated with phosphate-buffered saline (PBS; control), NGF (100 ng/mL) and/or DFN (0.25-1.00 mM) for 4 hours. We examined: (1) cell injury by confocal microscopy; (2) cell death/survival using Calcein AM live cell tracking dye; (3) mitochondrial structure and membrane potential function using MitoTracker in live cells; and (4) expression of NGF, its receptor - tropomyosin receptor kinase A (TrkA), survivin and IGF-1 by immunostaining. DFN treatment of RGM1 cells for 4 hours caused extensive cell injury, mitochondrial disintegration, reduced cell viability (from 94 ± 3% in controls to 14 ± 4% in 0.5 mM DFN-treated cells; P < 0.001), and expression of survivin and IGF-1. NGF treatment significantly increased survivin and IGF-1 expression by 41% and 75%, respectively versus PBS controls. Pretreatment with NGF before DFN treatment reduced mitochondrial damage and cell death by 73% and 82%, respectively versus treatment with DFN alone (all P < 0.001). This study also showed the presence of high-affinity TrkA receptors in the plasma membrane and mitochondria of RGM1 cells indicating novel actions of NGF. 相似文献
150.
Molecular dynamics of synthetic leucine-serine ion channels in a phospholipid membrane 总被引:2,自引:0,他引:2
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Molecular dynamics calculations were carried out on models of two synthetic leucine-serine ion channels: a tetrameric bundle with sequence (LSLLLSL)(3)NH(2) and a hexameric bundle with sequence (LSSLLSL)(3)NH(2). Each protein bundle is inserted in a palmitoyloleoylphosphatidylcholine bilayer membrane and solvated by simple point charge water molecules inside the pore and at both mouths. Both systems appear to be stable in the absence of an electric field during the 4 ns of molecular dynamics simulation. The water motion in the narrow pore of the four-helix bundle is highly restricted and may provide suitable conditions for proton transfer via a water wire mechanism. In the wider hexameric pore, the water diffuses much more slowly than in bulk but is still mobile. This, along with the dimensions of the pore, supports the observation that this peptide is selective for monovalent cations. Reasonable agreement of predicted conductances with experimentally determined values lends support to the validity of the simulations. 相似文献