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1.
Flow cytometry (FCM) in conjunction with immunocytochemical-labeling was used to analyze and screen a population of Escherichia coli clones containing a genomic library from the oil-degrading microorganism Acinetobacter calcoaceticus RAG-1 for the isolation of clones which expressed specific RAG-1 surface antigens. Reconstruction experiments using mixed populations indicated that RAG-1 cells could be clearly distinguished at a ratio of one RAG-1 cell to 500 Escherichia coli cells. Using this technique two clones, WM143 and WM191, were isolated and shown by restriction endonuclease cleavage and Southern hybridization to contain plasmids carrying inserts of RAG-1 DNA of 9.4 and 9.8 kb respectively.Non-common abbreviations FCM
flow cytometry
- FITC
fluorescein-iso-thiocyanate
- LB
Luria broth
- MM
minimal salt medium
- PBS
phosphate buffered saline
- PMSF
phenylmethylsulfonyl fluoride 相似文献
2.
Irene Konstantopoulou Christos A. Ouzounis Elena Drosopoulou Minas Yiangou Paschalis Sideras Chris Sander Zacharias G. Scouras 《Journal of molecular evolution》1995,41(4):414-420
A clone isolated from a Drosophila auraria heat-shock cDNA library presents two long, antiparallel, coupled (LAC) open reading frames (ORFs). One strand ORF is 1,929 nucleotides long and exhibits great identity (87.5% at the nucleotide level and 94% at the amino acid level) with the hsp70 gene copies of D. melanogaster, while the second strand ORF, in antiparallel in-frame register arrangement, is 1,839 nucleotides long and exhibits 32% identity with a putative, recently identified, NAD+-dependent glutamate dehydrogenase (NAD+-GDH). The overlap of the two ORFs is 1,824 nucleotides long. Computational analysis shows that this LAC ORF arrangement is conserved in other hsp70 loci in a wide range of organisms, raising questions about possible evolutionary benefits of such a peculiar genomic organization.Correspondence to: Z.G. Scouras 相似文献
3.
Wolfang Zeiske Heinz Marin 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1992,162(4):340-344
Summary Chloride-stimulated K+ secretion by Manduca sexta midgut (5th-instar larvae) was measured as K+-carried short-circuit current of the tissue mounted in an Ussing chamber. Microscopic parameters, such as single-channel current and channel density for the rate-determining passive transport step across the basolateral goblet cell membrane (i.e. K+ channels), were estimated by means of current-fluctuation analysis of the K+ channel blockade by haemolymph-side Ba2+ ions. Ba2+ was equally effective with Cl- or gluconate (Glu-) as the principal ambient anion. The Ba2+-induced K+ channel conduction noise is reflected by a Lorentzian, or relaxation, noise component in the power spectrum of the K+ current fluctuations. A reduced Lorentzian plateau value, but an unchanged corner frequency, were observed when Cl- was replaced by Glu-. The results from the analysis of a two-state model of K+ channel block by Ba2+, with respect to the anion-replacement effects, suggest that the observed changes in K+ current and Lorentzian plateau value mirror a complex change of the underlying parameters: Cl- omission reduces single channel current but increases channel density so that the product of single channel current and channel density is smaller in Glu- than in Cl-. It seems likely that basolateral K+ channels (1) are subject to anionic gating ligands, and (2) depend on anions with respect to the rate of K+ transfer through and open K+ channel.Abbreviations a.c.
alternating current
-
single-channel conductance
-
E
K
K+ Nernst potential
-
f
frequency contained in current noise
-
f
c
corner frequency
- Glu-
gluconate
-
G
t
transepithelial conductance
-
I
sc
short-circuit current
-
I
K
K+ current
-
I
K(max)
maximal K+ current
-
i
single-channel current
-
K
Ba
barium inhibition constant
-
K
m
Michaelis constant of saturating K+ current
-
k
01 and k
10
barium association and dissociation rate constant, respectively
-
M
K+ channel density
-
S
f
power density
-
S
o
Lorentzian plateau value
-
P
o
channel-open probability
-
P
K
K+ permeability
-
V
sc
cellular potential at short-circuit
These results have already been presented in part, at the 1989 joint meeting of the German and Israel Physiological Societies in Jerusalem (Zeiske et al. 1990). 相似文献
4.
Penelope Bouziotis Eleni Gourni George Patsis Dimitrios Psimadas Christos Zikos Melpomeni Fani Stavros Xanthopoulos George Loudos Maria Paravatou-Petsotas Evangelia Livaniou Alexandra D. Varvarigou Ioannis Pirmettis Minas Papadopoulos 《Bioorganic & medicinal chemistry》2013,21(21):6699-6707
Bombesin is a neuropeptide widely studied due to its ability to target various types of cancers. Technetium-99m on the other hand is ideal for diagnostic tumor targeting. The aim of the present study is the investigation of the coupling of the ligand (S)-(2-(2′-pyridyl)ethyl)-d,l-cysteine with the BN-peptide Gln-Arg-Leu-Gly-Asn-Gln-Trp-Ala-Val-Gly-His-Leu-Met(CONH2) through the spacer aminohexanoic acidand the labeling of the resulting derivative MBN with the synthon [M(CO)3(H2O)3]+ (M = 99mTc, Re). The peptide was synthesized according to the SPPS method, purified and characterized by ESI-MS. The new 99mTc-labeled biomolecule was stable in vitro, showed high affinity for the human GRP receptor expressed in PC3 cells and the rate of internalization was found to be time-dependent tissue distribution of the radiopeptide was evaluated in normal mice and in prostate cancer experimental models and significant radioactivity uptake was observed in the pancreas of normal mice as well as in PC3 tumors. Dynamic studies of the radiopeptide showed satisfactory tumor images. 相似文献
5.
6.
Minas Papadopoulos Berthold Nock Theodosia Maina Ioannis Pirmettis Catherine Raptopoulou Anastasios Tasiopoulos Anastasios Troganis Themistoklis Kabanos Aris Terzis Efstratios Chiotellis 《Journal of biological inorganic chemistry》2001,6(2):159-165
Novel oxorhenium and oxotechnetium complexes based on the tetradentate 1-(2-hydroxybenzamido)-2-(pyridinecarboxamido)benzene, H3L, ligand have been synthesized and characterized herein. Thus, by reacting equimolar quantities of the triply deprotonated ligand L3- with the suitable MO3+ precursor, the following neutral MOL complexes could be easily produced following similar synthetic routes: M = Re (1), M = 99gTc (2), and M = 99mTc (3). Complexes 1 and 2, prepared in macroscopic amounts, were chemically characterized and their structure determined by single-crystal X-ray analysis. They are isostructural metal chelates, adopting a distorted square pyramidal geometry around the metal. The N3O donor atom set of the tetradentate ligand defines the basal plane and the oxygen atom of the M = O core occupies the apex of the pyramid. Complex 3 forms quantitatively at tracer level by mixing the H3L ligand with Na99mTcO4 generator eluate in aqueous alkaline media and using tin chloride as reductant in the presence of citrate. Its structure was established by chromatographic comparison with prototypic complexes 1 and 2 using high-performance liquid chromatographic techniques. When challenged with excess glutathione in vitro, complex 3 is rapidly converted to hydrophilic unidentified metal species. Tissue distribution data after administration of complex 3 in vivo revealed a significant uptake and retention of this compound in brain tissue. 相似文献
7.
Chrysopoulo MT McGrouther DA Jeschke MG Kaufman BR 《Plastic and reconstructive surgery》2002,109(2):566-72; discussion 573-5
8.
9.
Adiponectin induces TNF-alpha and IL-6 in macrophages and promotes tolerance to itself and other pro-inflammatory stimuli 总被引:7,自引:0,他引:7
Tsatsanis C Zacharioudaki V Androulidaki A Dermitzaki E Charalampopoulos I Minas V Gravanis A Margioris AN 《Biochemical and biophysical research communications》2005,335(4):1254-1263
Adiponectin exerts anti-inflammatory effects via macrophages, suppressing the production of pro-inflammatory cytokines in response to bacterial lipopolysaccharide (LPS). Here, we provide experimental evidence that the "anti-inflammatory" effect of adiponectin may be due to an induction of macrophage tolerance: globular adiponectin (gAd) is a powerful inducer of TNF-alpha and IL-6 secretion in primary human peripheral macrophages, in the THP-1 human macrophage cell line, and in primary mouse peritoneal macrophages. Pre-exposure of macrophages to 10 microg/ml gAd rendered them tolerant to further gAd exposure or to other pro-inflammatory stimuli such as TLR3 ligand polyI:C and TLR4 ligand LPS, while pre-exposure to 1 microg/ml of and re-exposure to 10 microg/ml gAd unmasked its pro-inflammatory properties. GAd induced NF-kappaB activation and tolerance to further gAd or LPS exposure. Our data suggest that adiponectin constant presence in the circulation in high levels (in lean subjects) renders macrophages resistant to pro-inflammatory stimuli, including its own. 相似文献
10.
Ochoa O Chrysopoulo M Nastala C Ledoux P Pisano S 《Plastic and reconstructive surgery》2012,130(1):21e-33e