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Query: EC:3.1.27.1 (
RNase
)
16,360
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Along with classical
lipopolysaccharide
(
LPS
), O-specific material not precipitated by ultracentrifugation has been isolated from the water-phenol extract of S. sonnei avirulent strain 9090 possessing complete antigenic properties. The purification of O-antigen contained in the supernatant fluid has been carried out by the gel filtration of the fluid, previously treated with
ribonuclease
, in a column packed with Sephadex G-100. The polysaccharide nature of O-antigen thus obtained, the absence of lipid A and KDO and the low content of hexoses, or core-specific saccharides of S. sonnei
LPS
, in this antigen make it possible to classify this material with O-components of microbial cells, described by different authors as "native protoplasmic polysaccharide" or "L-hapten" and formed by polymers of
LPS
O-side chains. The content of this component in S. sonnei strains under study is, on the average, 2.5% of the weight of dry microbial substance. L-hapten preparations obtained in the course of our investigations have been found to contain two O-specific antigens detected by immunoelectrophoresis and immunodiffusion, as well as by sedimentation in saccharose gradient, where they form peaks corresponding to 4.3 S and 10.8 S. This polysaccharide O-antigen is supposed to be capable of interaction with ribosomal particles and suitable for use as a component of ribosomal dysentery vaccines.
...
PMID:[O-specific L-hapten in the composition of Shigella sonnei]. 248 42
Interleukin 4 (IL-4) induces the expression of IgG1 and IgE in
lipopolysaccharide
-stimulated B cells. Previous studies have suggested that heavy-chain class switching may be regulated by increasing the accessibility of specific switch regions to switch recombinases. In this study, we have used an
RNase
protection assay to demonstrate that IL-4 induces expression of germ-line gamma 1 transcripts in B cells within 4 hr of culture; induction is dose-dependent and is inhibited by interferon gamma. IL-4 alone is capable of inducing the expression of germ-line gamma 1 transcripts in small, resting B cells, but
lipopolysaccharide
enhances expression. The germ-line transcripts are the same size (1.8 and 3.4 kilobases) as the secreted and membrane forms of the functional gamma 1 mRNAs and presumably result from the splicing of an upstream switch-region exon(s) to the gamma 1 constant-region exon(s). These data strongly support the "accessibility" model for the regulation of isotype switching and suggest that lymphokines such as IL-4 may direct specific switch events by transcriptional activation of the corresponding switch regions.
...
PMID:Synthesis of germ-line gamma 1 immunoglobulin heavy-chain transcripts in resting B cells: induction by interleukin 4 and inhibition by interferon gamma. 249 37
Tumor necrosis factor (TNF) and interleukin-1 (IL-1) play an intimate role in the initiation and maintenance of inflammatory reactions due to their pluripotent activities. In this paper, we describe the use of an in situ hybridization analysis as an effective means to probe for TNF and IL-1 mRNA levels in primary macrophage cultures and macrophage cell lines. A significant increase in
lipopolysaccharide
(
LPS
)-induced TNF mRNA accumulation was demonstrated by in situ hybridization using either a 35S-labeled synthetic oligonucleotide (30-mer) complementary to TNF mRNA or a 35S-randomly primed labeled TNF DNA probe. An augmentation in TNF mRNA accumulation, as assessed by increasing grains/cell, was demonstrated over a wide concentration range of
LPS
. This accumulation was shown using both immunologically elicited primary macrophage cultures and the macrophage cell line RAW 264.7. Interestingly, the RAW 264.7 constitutively produced TNF in the absence of specific stimulus and this tonic production was observed at the molecular level via in situ hybridization analysis. Specificity of the in situ hybridization technique was shown by a complete loss in binding of 35S-probe after either
RNase
digestion or competition with "cold-labeled" probe. beta-actin served as a 35S-labeled control probe where the number of actin-specific grains/cell was not altered by stimulating macrophages with
LPS
. IL-1 alpha mRNA was also increased by
LPS
stimulation of macrophages as assessed by in situ hybridization. The
LPS
-dependent increase in macrophage mRNA for TNF and IL-1 alpha, as assessed by in situ hybridization, was confirmed by classical Northern blot analysis as well as the production of biologically-active protein.
...
PMID:In situ hybridization analysis of macrophage-derived tumor necrosis factor and interleukin-1 mRNA. 326 57
To detect nuclear proteins that might be involved in induction of cellular mitogenesis, we examined the effect of various mitogens on early changes in synthesis of nuclear proteins in murine B lymphocytes. Using two-dimensional gel electrophoresis, we found that activation of B cells by mitogens (anti-immunoglobulin antibody,
lipopolysaccharide
, phorbol 12-myristate 13-acetate (PMA)/A23187) was associated with a rapid and prominent (5-20-fold) increase in the synthesis of a 40-kDa/pI 5.0 nuclear protein, here termed numatrin. Numatrin was found to be absent from the cytosol (soluble fraction) of resting as well as activated B cells and was markedly resistant to DNase/
RNase
digestion and 2 N NaCl extraction, indicating that this protein is tightly bound to the nuclear matrix. Kinetic studies showed that the increase in synthesis of numatrin was detected 60-120 min following mitogen activation, reached a peak at 16 h, and declined to almost control level by 48 h, correlating with the peak of cellular DNA synthesis. The increase in synthesis of numatrin in normal B cells was found to be associated exclusively with cellular commitment for mitogenesis because activation of B cells by stimuli such as B cell stimulating factor 1, PMA alone, and calcium ionophore A23187, which do not stimulate an increase in DNA synthesis, also failed to induce an increase in the synthesis of numatrin. Inhibition of anti-Ig-induced proliferation (by PMA pretreatment) was associated with a 63% inhibition in the synthesis of numatrin. Addition of 8-mercaptoguanosine to these PMA-treated cells was associated with restoration of the increase in synthesis of numatrin, concomitant with induction of proliferation. Elevated synthesis of numatrin was also detected in the malignant B lymphoma cells: Raji, BAL-17, and WEHI-231. Taken collectively, these results suggest that numatrin, a tightly bound nuclear matrix protein, is a growth-regulated protein which might have an important role in regulation of cellular mitogenesis in normal and malignant B lymphocytes.
...
PMID:"Numatrin," a nuclear matrix protein associated with induction of proliferation in B lymphocytes. 330 55
A new approach has been developed to circumvent the problems of false positive reactions in the Limulus Amoebocyte Lysate (LAL) assay for
lipopolysaccharide
(
LPS
) in root surface materials. These LAL-reactive materials include thrombin, thromboplastin,
ribonuclease
, ribonucleic acid, lipoteichoic acid and peptidoglycan fragments. In the present study, hot phenol/water extraction of these substances followed by ultracentrifugation of the resulting aqueous phases reduced their concentrations to very low levels. Furthermore, the application of Polymyxin B/Sepharose 4B affinity chromatography to these extracts enabled their intrinsic LAL-activity to be determined. Use of these techniques to assay root surface materials has identified
LPS
as being the major LAL-reactive material present. The mean
LPS
yield for the periodontally involved teeth was 4.13 micrograms/tooth, representing 2.82 micrograms/root. In contrast, the mean yield of
LPS
for the periodontally uninvolved teeth was 3.12 ng/tooth.
...
PMID:Identity of limulus amoebocyte lysate-active root surface materials from periodontally involved teeth. 346 18
Various uniform salt forms of an R-form
lipopolysaccharide
(
LPS
) extracted from Klebsiella strain LEN-111 (O3-:K1-) were prepared and their ultrastructure was examined. The
LPS
, which was extracted by the phenol-water method, freed from contamination with RNA by treatment with
RNase
, and precipitated by addition of two volumes of 10 mM MgCl2-ethanol, was used as the original preparation for uniform salt forms. The original
LPS
preparation formed a hexagonal lattice structure with a lattice constant of 14.9 +/- 0.2 nm. The
LPS
after electrodialysis retained the ability to form a hexagonal lattice structure, although its lattice constant was large (18.7 +/- 0.5 nm) and the lattice structure of the electrodialyzed
LPS
was labile at pH 8.0 in contrast to that of the original
LPS
preparation. The magnesium salt form of the
LPS
formed essentially the same ordered hexagonal lattice structure (lattice constant of 15.0 +/- 0.2 nm) as that of the original
LPS
preparation. The calcium and ammonium salt forms formed a hexagonal lattice structure, but the lattice constants of the calcium and ammonium salt forms were larger (18.6 +/- 0.6 nm and 19.3 +/- 0.4 nm, respectively) than that of the magnesium salt form. The sodium and potassium salt forms consisted of freely branching ribbon-like structures with an average width of 13 nm and an average thickness of 9 nm. The triethylamine salt form consisted principally of short rods (10 nm X 9-13 nm).
...
PMID:Formation of a hexagonal lattice structure by an R-form lipopolysaccharide of Klebsiella: relationship between lattice formation and uniform salt forms. 409 71
A
lipopolysaccharide
(
LPS
) fraction was extracted from Nichols, nonpathogenic Treponema pallidum by the hot, phenol-water procedure. The
LPS
was freed of nucleic acids and water-soluble proteins by successive exposures to
ribonuclease
, deoxyribonuclease, and Pronase. Purified
LPS
responded positively in a colorimetric assay for
lipopolysaccharide
. Electron microscope examination of the
LPS
both before and after purification demonstrated a heterogeneous mixture of forms including spheres, doughnuts, and ribbons. The trilaminar nature of the ribbon forms was observed by both negative staining and thin sectioning. Lyophilization of the
LPS
caused an increase in the number and length of ribbon forms seen. Results suggest that the surface layers of treponemes are similar to those of gram-negative bacteria.
...
PMID:Ultrastructure of lipopolysaccharide isolated from Treponema pallidum. 412 67
In mice, active protection against Pseudomonas aeruginosa could be induced with two fractions derived from a crude preparation of ribosomes from P. aeruginosa. The two fractions were obtained by gel filtration chromatography of the crude ribosomal preparation on Sepharose CL-2B. In fraction I, less than 1% of the ribonucleic acid (RNA) applied to the column was recovered. Fraction II contained RNA and protein in a ratio of 1.94. The presence of ribosomes in this fraction was confirmed by analysis on a sucrose density gradient. The protection by fraction I was not affected by treatment with
ribonuclease
; in contrast, incubation of fraction II with
ribonuclease
completely abolished active protection. Fraction I contained
lipopolysaccharide
(
LPS
) as was indicated by the presence of 2-keto-3-deoxyoctonic acid. No
LPS
was found in fraction II. The adjuvant dimethyl dioctadecyl ammonium bromide enhanced the protection by fraction II; however, immunity by a low dose of fraction I was abolished by dimethyl dioctadecyl ammonium bromide. Protection by fractions I and II appeared to be restricted to the homologous serotype of P. aeruginosa. These results indicate that RNA is required for protection by fraction II. Active protection by fraction I is likely due to
LPS
.
...
PMID:Ribonuclease-sensitive ribosomal vaccine of Pseudomonas aeruginosa. 615 37
Ribosomal and ribonucleic acid (RNA)-rich preparations derived from Salmonella typhimurium were examined for their ability to enhance the primary in vitro antibody response of normal mouse spleen cell cultures to sheep erythrocytes. Both of these fractions were consistently more active in elevating the antibody response of normal mouse splenocytes from
lipopolysaccharide
(
LPS
) responder mice than was
LPS
. Furthermore, injection of mice with either the ribosomal or RNA-rich fraction induced antibody response helper factor activity in 2-h post-treatment serum similar to that induced by
LPS
. Endotoxin low-responding C3H/HeJ mice were stimulated to release helper factors by ribosomes and the RNA extracts but not by
LPS
. Treatment of the ribosomes and RNA fractions with
ribonuclease
destroyed their ability to stimulate the production of the helper factor in serum of treated mice. Therefore, it appears likely that ribosomes and RNA-rich fractions stimulated an intermediate helper factor due to the presence of RNA and not
LPS
.
...
PMID:Induction of immunoenhancing factors for murine splenocyte cultures by Salmonella typhimurium ribosome and ribonucleic acid extracts. 616 65
To obtain information about the nature of the immunogens in the ribosomal vaccine (fraction II) of Pseudomonas aeruginosa, we studied the specificity of rabbit antibodies to fraction II. Crossed immunoelectrophoresis demonstrated the presence of antibodies which precipitated with ribosomal antigens, but not with
lipopolysaccharide
(
LPS
). By means of an enzyme-linked immunosorbent assay, antibodies to
LPS
were detected in antibodies to fraction II. Antibodies to fraction II could protect mice against a lethal challenge with P. aeruginosa. Absorption experiments demonstrated that the protective ability of antibodies to fraction II was due to antibodies to cell envelope antigens, whereas antibodies to ribosomal antigens did not contribute to the protection. Antibodies to
LPS
could be detected in mice 1 week after a single vaccination with fraction II. It was concluded that the protective activity of fraction II was due, at least in part, to the presence of
LPS
in the ribosomal vaccine. Treatment of fraction II with
ribonuclease
decreased the protective activity of the ribosomal vaccine. Addition of synthetic polyadenylic acid-polyuridylic acid restored the protective activity of
ribonuclease
-treated fraction II, indicating that RNA in the ribosomal vaccine might act as an adjuvant or a carrier in the presentation of the of the contaminating cell envelope antigens. The protective activity and the toxicity of fraction II were compared with the protective activity and the toxicity of fraction I, which contained cell envelope components, including
LPS
, and of purified
LPS
. The results indicated that protection by the ribosomal vaccine was associated with a slightly higher toxicity than was protection by fraction I, whereas purified
LPS
was the most toxic vaccine.
...
PMID:Evidence for the presence of lipopolysaccharide in a ribonuclease-sensitive ribosomal vaccine of Pseudomonas aeruginosa. 678 42
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