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41.
Radioimmunoassay for human type VI collagen and its application to tissue and body fluids 总被引:3,自引:0,他引:3
A liquid phase radioimmunoassay (RIA) was developed for pepsin-solubilized human type VI collagen, allowing quantitative analysis of this protein down to a concentration of 3 ng/ml. No cross-reactivity was observed with human collagens type I, III, IV (triple helical portion and 7-S domain), and V, nor with laminin fragment Pl and plasma fibronectin. Significant amounts of closely related antigenic material were detected in serum, bile, ascites, and mesenchymal cell culture media. Type VI collagen could be completely solubilized from several tissues by a repeated pepsin digest, and its content as determined by RIA was found to be less than 0.1% of total collagen (55-70 micrograms/g protein). In fibrotic liver tissue type VI collagen was elevated up to 10-fold (620 micrograms/g protein) when compared to normal liver. Sera of patients with fibrotic liver disease, however, revealed antigen levels usually below the narrow normal range of 22 +/- 7.8 ng/ml (mean +/- 2.5 SD). We conclude that, although type VI collagen represents a minor fraction of the interstitial collagens, its comparatively high serum levels point to a considerable turnover in the normal individual. Our data suggest that in fibrosis as exemplified in fibrotic liver disease, the metabolism of this collagen is down-regulated, while at the same time, it accumulates in the interstitial matrix. 相似文献
42.
43.
Nucleotide sequence analysis of a variant human T-cell leukemia virus (HTLV-Ib) provirus with a deletion in pX-I. 总被引:11,自引:9,他引:2
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L Ratner S F Josephs B Starcich B Hahn G M Shaw R C Gallo F Wong-Staal 《Journal of virology》1985,54(3):781-790
A variant of human T-cell leukemia virus subgroup I (HTLV-I), designated HTLV-Ib, has been isolated from a transformed T-lymphocytic cell line established from a Zairian patient with adult T-cell lymphoma. A recombinant phage clone of the variant provirus, denoted lambda MC-1, hybridizes under high stringency to HTLV-I DNA probes, but 17 of 43 restriction enzyme sites differ from those of HTLV-I, 10 of them clustering within 1.5 kilobases in the env-pX region. Since this variant virus retains its capacity to transform T-cells in vitro, and since a pX product is suspected to be important in transformation, we have determined the nucleotide sequence of the entire pX region of this virus for comparison to the prototype HTLV-I. In addition, the region between the gag and pol genes, parts of the pol and env genes, and a portion of the U3 region of the long terminal repeat sequence were also analyzed. We noted 141 single-base-pair changes among 3,897 base pairs, which were relatively well distributed over those portions of the provirus that were examined. In addition, an 11-base-pair deletion was found which included the potential initiator ATG codon of the first open reading frame of pX (pX-I). The next potential initiator codon predicted by the sequence is followed by 10 codons and then a termination codon. An identical deletion was also demonstrated in the only provirus present in another cell line established from the same patient on a different occasion after transformation in vitro of normal human umbilical cord blood cells. These results indicate that pX-I is not required for transformation. 相似文献
44.
K. D. Hahn F. Buck H. Rüterjans B. K. Chernov K. G. Skryabin M. P. Kirpichnikov 《European biophysics journal : EBJ》1985,12(2):87-95
The 17 base pair operator O
R
3 oligonucleotide, which is the preferential binding site for the Cro repressor of phage , was studied by two-dimensional NMR spectroscopy. A sequential assignment procedure based on two-dimensional Nuclear Overhauser Effect (NOESY) and scalar coupling correlated (COSY) NMR spectroscopy, together with the knowledge of the oligodesoxynucleotide sequence, made it possible to assign the non-exhangeable base protons and the H1 and the H2-H2 sugar protons of the O
R
3 operator DNA. The pattern of the observed NOE connectivities is consistent with a right-handed helical DNA structure. The base and sugar proton assignments provide the necessary information for further studies of the O
R
3 operator — Cro repressor interaction.Abbreviations COSY
correlated spectroscopy
- FID
free induction decay
- NOE
nuclear Overhauser effect
- NOESY
nuclear Overhauser effect spectroscopy
- RD
relaxation delay
- TSP
sodium 3-trimethylsilyl-(2,2,3,3-2H4)propionate
- EDTA
sodium ethylendiamine tetraacetate 相似文献
45.
The potential use of multiple-frequency-band radiometry as a means of noninvasive sensing of one-dimensional temperature profiles is presented in this communication. The radiative energy transfer equation is solved numerically. Ideal-condition thermal noise spectra and distributions of received energy, associated with specific temperature-depth profiles, are presented. Performance characteristics are discussed. 相似文献
46.
The fine structure of protonephridia in Gnathostomulida and their comparison within Bilateria 总被引:1,自引:0,他引:1
Volker Lammert 《Zoomorphology》1985,105(5):308-316
Summary The fine structure of the protonephridia of Haplognathia rosea (Filospermoidea) and Gnathostomula paradoxa (Bursovaginoidea) is described. Each protonephridium consists of three different cells: (1) a monociliated terminal cell which constitutes the filtration area, (2) a nonciliated canal cell showing a special protonephridial outlet system, and (3) an intraepidermal cell — the nephroporus cell — constituting the nephroporus. The protonephridia are arranged serially. There is no canal system connecting the protonephridial units.Protonephridial characters in other Bilateria are considered. The pattern of characters in the protonephridia in the last common gnathostomulid stem species and presumed apomorphies in the protonephridia of the Gnathostomulida investigated are discussed.Abbreviations used in figures
ac
acessory centriole
-
AC
additional epidermal cell
-
bb
basal body
-
bl
basal lamina
-
bm
bundle of microvilli
-
c
cilium
-
cc
cilium duct cell
-
cd
cilium duct
-
cr
ciliary rootlet
-
crs
structures resembling ciliary rootlets
-
di
diplosome
-
ds
desmosome
-
dy
dictyosome
-
f
filtration area
-
g
granules
-
m
mitochondrium
-
mv
microvillus
-
n
nucleus
-
NC
nephroporus cell
-
np
nephroporus
-
oc
outlet canal
-
TC
terminal cell
-
tl
tubules of lacunar system 相似文献
47.
Thomas Hansen-Hagge Volker Lehmann Ulrich Seydel Buko Lindner Ulrich Zähringer 《Archives of microbiology》1985,141(4):353-358
The extraction, purification and structural characterization of two lipid A precursors (Ia and Ib) differing only in one hexadecanoic acid are described. Both precursors were synthesized at elevated temperatures by a new mutant of Salmonella typhimurium (mutant Ts5) which is conditionally defective in synthesis of the 3-deoxy-d-manno-octulosonic acid region of lipopolysaccharides.Both precursors were purified by repeated phenol/chloroform/petroleum ether (PCP) extractions followed by thin layer chromatography. Teh precursor preparation was free of lipopolysaccharides and phospholipids and contained less than 0.1% protein. Structural analysis which included chemical degradation procedures as well as positive ion laser desorption (LDMS) mass spectroscopy of dephosphorylated lipid A precursors showed together that precursor Ia represents a diphosphorylated glucosamine disaccharide containing two ester, two amide-linked residues of 3-hydroxytetradecanoic acid and lacks the ester-linked dodecanoic, tetradecanoic and hexadecanoic acid as well as 3-deoxy-d-manno-octulosonic acid. Precursor Ib has the same basic structure as precursor Ia, but contains in addition one mol of hexadecanoic acid per mol disaccharide which is linked to the 3-hydroxy group of the amide-bound 3-hydroxy-tetradecanoic acid of the reducing, terminal glucosamine residue.The structure of precursor Ib supports the conclusion that hexadecanoic acid incorporation occurs at an early stage in lipid A biosynthesis prior to the attachment of 3-deoxy-d-manno-octulosonic acid and/or other polar substituents.Abbreviations LDMS
laser desorption mass spectrometry
- KDO
3-Deoxy-d-manno-octulosonic acid
- Ts5
Salmonella typhimurium mutant Ts5
- PCP
phenol/chloroform/petroleum ether
- H2F2
hydrogen fluoride
This work is dedicated to Prof. Dr. Drews, Freiburg, on the occasion of his 60th birthday 相似文献
48.
Prostaglandin (PG) production was evaluated in the three cell types (endothelial, smooth muscle, and fibroblast) comprising the bovine pulmonary artery. Prostacyclin (PGI2) was the predominant prostaglandin (PG) produced by endothelial, smooth muscle, and fibroblast cells as they exist in culture or in freshly excised tissue fragments. In addition to PGI2, measurable amounts of PGE2, PGF2a, and thromboxane A2 (TXA2) were also produced by these cells. Endothelial cells were the most active producers of PGs. However, the type of PG produced was characteristic of the particular cell type, while the level of production was dependent on external factors. Prostaglandin production by cultured cells, both under basal conditions and in response to stimulatory agents, was quite similar to that of the respective freshly excised tissue fragments containing a given cell type. These cells in culture could be stimulated to produce PGI2 by both angiotensin and bradykinin at very low (physiological) concentrations, a further indication of the retention of the physiological responsiveness of these cells in culture. Endothelial cells and fibroblasts were activated by bradykinin at concentrations as low as 10(-12) M but did not respond to angiotensin. Smooth muscle cells in primary and first passage cultures were activated by both bradykinin and angiotensin at 10(-12) M concentrations. Serial subcultivations of smooth muscle cells resulted in a progressive loss in their responsiveness to bradykinin stimulation. The state of cell growth proved to be an important determinant of PG production. Actively growing cells in culture synthesized less PG when compared to cells which had entered into a "quiescent" nongrowth state. 相似文献
49.
We exposed two awake dogs with a chronic tracheostomy and the cervical vagus nerves exteriorized in skin loops to 1.0 ppm of ozone (O3) for 2 h at intervals of 4 wk. We measured ventilatory variables before and after O3 exposure during rest and exercise before and after vagal block. We compared the effects of vagal blockade, exercise, and O3 on the primary determinants of breathing pattern (VT/TI, VT/TE, TI, and TE) in each of three conditions: base line (steady state), during hypercapnia, and after inhalation of 1% histamine. Under base-line conditions, O3 increased respiratory rate and decreased tidal volume (VT) by shortening time of expiration (TE) and time of inspiration (TI) without affecting VT/TI, an indicator of the neural drive to breathing. During progressive hypercapnia, O3 shortened TE and TI by effects both on tonic (nonvolume-related) and on phasic (volume-related) vagal inputs, and only the latter were prevented completely by cooling of the vagus nerves. Histamine-induced tachypnea was increased by O3 and was totally blocked by cooling the vagus nerves. We conclude that O3 shortens the timing of respiration without increasing ventilatory drive, shortens TI and TE through vagal and nonvagal pathways, increases tonic nonvagal and phasic vagal inputs, and stimulates more than one vagal fiber type. 相似文献
50.
B. Regensburger L. Meyer M. Filser J. Weber D. Studer J. W. Lamb H. -M. Fischer M. Hahn H. Hennecke 《Archives of microbiology》1986,144(4):355-366
The genome of the slow-growing Bradyrhizobium japonicum (strain 110) was mutagenized with transposon Tn5. A total of 1623 kanamycin/streptomycin resistant derivatives were screened in soybean infection tests for nodulation (Nod) and symbiotic nitrogen fixation (Fix). In this report we describe 14 strains possessing a stable, reproducible Nod+Fix- phenotype. These strains were also grown under microaerobic culture conditions to test them for free-living nitrogen fixation activity (Nif). In addition to strains having reduced Fix and Nif activities, there were also strains that had reduced symbiotic Fix activity but were Nif+
ex planta.Analysis of the genomic structure revealed that the majority of the strains had a single Tn5 insertion without any further apparent physical alteration. A few strains had additional insertions (by Tn5 or IS50), or a deletion, or had cointegrated part of the vector used for Tn5 mutagenesis. One of the insertions was found in a known nif gene (nifD) whereas all other mutations seem to affect different, hitherto unknown genes or operons.Several mutant strains had an altered nodulation phenotype, inducing numerous, small, widely distributed nodules. Light and electron microscopy revealed that most of these mutants were defective in different stages of bacteroid development and/or bacteroid persistence. The protein patterns of the mutants were inspected by two-dimensional gel electrophoresis after labelling microaerobic cultures with l-(35S)methionine. Of particular interest were mutants lacking a group of proteins the synthesis of which was known to be under oxygen control. Such strains can be regarded as potential regulatory mutants. 相似文献