ACTH-like bioactivity and immunoactivity in fetal lamb pituitaries at 0.65 and 0.95 gestation

1996 ◽  
Vol 8 (1) ◽  
pp. 195 ◽  
Author(s):  
MA Fora ◽  
NK Valego ◽  
MO Lively ◽  
MI Castro ◽  
JC Rose
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Late Gestation ◽  
Size Exclusion ◽  
Molecular Forms ◽  
Low Molecular Weight ◽  
High Molecular Weight Fraction ◽  
Crude Extracts ◽  
Molecular Weight Peak ◽  
Ovine Fetal

We wished to determine if the concentration of bioactive ACTH-like activity increased during development and if there was heterogeneity in ovine fetal anterior pituitary ACTH activity as measured by bioassay and radioimmunoassay (RIA). We obtained anterior pituitaries from eight sheep fetuses (four at 0.65 and four at 0.95 gestation; term 145 +/- 5 days) and extracted and homogenized them in ice-cold 5N acetic acid, 0.3% phenylmethanesulfonyl fluoride (PMSF) and 0.2% BSA. Fractionation of each pituitary extract was performed by size-exclusion chromatography using Sepadex G-50. The ACTH-like immunoactivity (ALI) profile for each pituitary showed two well-defined peaks. One eluted with human ACTH1-39 and the other eluted with the high molecular weight fraction in the void volume. Four fractions from the first peak representing the high molecular weight forms of ACTH activity and four fractions from the second peak representing the low molecular weight forms of ACTH activity were pooled separately. These two pools were subjected to reverse-phase chromatography (RPC) on a C-8 column using a linear gradient of 70% acetonitrile in 0.8% trifluoroacetic acid over a 60 min period. Based upon the RIA, the high molecular weight forms of ACTH from the G-50 column were resolved into three main fractions, one eluting similar to the standard ACTH1-39 and the remaining two eluting after that. The low molecular weight forms of ACTH from the G-50 column were resolved into three peaks, before, with, and after the standard. We used collagenase-dispersed rat adrenal cells to test the ACTH-like bioactivity (ALB) of the crude extracts and of the different fractions obtained from the RPC of the high and low molecular weight material. The concentration of ACTH-like bioactivity in the crude extracts was similar at the two stages of gestation. However, there was a trend for the low molecular weight peak to have more peptide eluting with human ACTH1-39 and higher ratios of ALB/ALI than did the high molecular weight peak. These results suggest that multiple ACTH molecular forms with different ALB/ALI ratios are present in the ovine fetal pituitary and that there is no selective increase in ACTH1-39 concentration in the fetal pituitary in late gestation.

Antithrombotic and Anticoagulant Activities of a Low Molecular Weight Fucoidan by the Subcutaneous Route

1999 ◽  
Vol 81 (03) ◽  
pp. 391-395 ◽  
Author(s):  
S. Colliec Jouault ◽  
S. Mauray ◽  
J. Theveniaux ◽  
C. Sternberg ◽  
Boisson Vidal ◽  
...  
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Subcutaneous Injection ◽  
Thrombin Generation ◽  
Sulfated Polysaccharides ◽  
Low Molecular Weight ◽  
High Molecular Weight Fraction ◽  
Antithrombotic Activity ◽  
Single Subcutaneous Injection ◽  
Chemical Structures

SummaryFucoidans (high-molecular-weight sulfated polysaccharides extracted from brown seaweeds) have anticoagulant and antithrombotic effects. They inhibit thrombin by catalyzing both serpins (antithrombin and heparin cofactor II) according to their chemical structures and origins. In this study, a low-molecular-weight (LMW) fucoidan of 8 kDa was obtained by chemical degradation of a high-molecular-weight fraction. The antithrombotic and anticoagulant activities of this new compound were compared to those of a low-molecular-weight heparin (LMWH), dalteparin, following subcutaneous administration to rabbits. This LMW fucoidan exhibited dose-related venous antithrombotic activity, with an ED80 of about 20 mg/kg, 2 h after a single subcutaneous injection. Its activity was comparable to that of dalteparin (close to 200 anti-Xa IU/kg) and was maximal 30 min after a single subcutaneous injection. The activity remained stable (about 70%) from 1 to 4 h after injection, but disappeared by 8 h. The antithrombotic activity was not associated with either a prolongation of the thrombin clotting time (TCT) or an increase in anti-Xa activity, contrary to dalteparin. A slight prolongation of APTT occurred with both compounds. This venous antithrombotic activity was associated with a decrease in ex vivo thrombin generation and with a significant increase in the lag phase in a thrombin generation test. LMW fucoidan thus has potent antithrombotic activity and a potentially weaker haemorrhagic effect (i.e. a smaller effect on coagulation tests and a smaller prolongation of the bleeding time) than dalteparin.

FRACTIONATION OF LIVER SOMATOMEDIN ACTIVITY BY ULTRAFILTRATION

1974 ◽  
Vol 62 (2) ◽  
pp. 355-361 ◽  
Author(s):  
JENNIFER M. DEHNEL ◽  
P. D. McCONAGHEY ◽  
M. J. O. FRANCIS
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Weight Fraction ◽  
Low Molecular Weight ◽  
High Molecular Weight Fraction ◽  
Cartilage Growth ◽  
The Relationship ◽  
Somatomedin Inhibitors

SUMMARY Plasma somatomedin is the intermediary through which growth hormone (GH) exerts its effects on the growing skeleton. Somatomedin activity may be produced in vitro by perfusion of the liver and kidneys of rats with Waymouth's medium containing GH. The relationship between the activity of plasma somatomedin and somatomedin of hepatic and renal origin has yet to be clarified. Somatomedin from plasma can be separated into active fractions of both high and low molecular weight. Similarly, ultrafiltration of medium containing somatomedin of hepatic origin indicates the existence of two active fractions, one of high molecular weight (greater than 50000) and one of low molecular weight (less than 1000). The latter can be attributed to the release of amino acids, such as serine and glutamine, by the perfused tissue. The high molecular weight fraction is believed to represent GH-dependent somatomedin. Fractions that inhibit production of cartilage matrix are present in liver perfusates as well as in plasma. These results provide further evidence that the liver is a source of GH-dependent somatomedin in vivo. Furthermore, cartilage growth may be controlled not only by the GH-stimulated release of somatomedin by the liver, but also by its release of acid-labile somatomedin inhibitors.

Distribution of radioactive silver in the subcellular fractions of various tissues of the rat and its binding to low molecular weight proteins

1983 ◽  
Vol 61 (11) ◽  
pp. 1391-1395 ◽  
Author(s):  
Yousef Matuk
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Gel Filtration ◽  
Total Radioactivity ◽  
Subcellular Fractions ◽  
Low Molecular Weight ◽  
Electron Microscopic ◽  
Molecular Weight Peak

In view of the electron microscopic evidence that silver does not penetrate cellular barriers, the distribution of radioactive silver in rat blood and subcellular fractions of liver, kidneys, spleen, and forebrain was studied. It was found that 24 h after a single intraperitoneal injection high levels of radioactivity were reached which decreased at different rates in the various tissues studied. In plasma, liver, and kidneys there was an initial rapid loss of radioactivity which was followed by a slower rate of loss. In the blood, forebrain, and spleen the loss of radioactivity was linear and somewhat slower than in the other three tissues. The cytosols of the liver and kidneys contained 60% while those of the forebrain and spleen contained 30% of the total radioactivity found in the tissue homogenates. Gel filtration on Sephadex G-75 showed that all cytosols contained two peaks of radioactivity; a high molecular weight peak which eluted just after the void volume and a low molecular weight peak. The amount of radioactivity in both peaks was, however, much lower in the chromatographic peaks of the forebrain and spleen than that found in those of the liver and kidneys. Furthermore, the spleen had a comparatively very small low molecular weight radioactive peak. In vitro experiments with liver cytosol showed similar results to those found in vivo in that the high molecular weight radioactive peak could be removed by heat. It is concluded that silver does enter cells and that silver thionein exists in the cytosols of forebrain, spleen, kidney, and liver.

scholarly journals Exopolysaccharide Production and Ropy Phenotype Are Determined by Two Gene Clusters in Putative Probiotic Strain Lactobacillus paraplantarum BGCG11

2014 ◽  
Vol 81 (4) ◽  
pp. 1387-1396 ◽  
Author(s):  
Milica Zivkovic ◽  
Marija Miljkovic ◽  
Patricia Ruas-Madiedo ◽  
Ivana Strahinic ◽  
Maja Tolinacki ◽  
...  
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Gene Cluster ◽  
Probiotic Strain ◽  
Gene Clusters ◽  
Size Exclusion ◽  
High Molecular Weight Fraction ◽  
Transposase Gene ◽  
Content Type ◽  
Lactobacillus Paraplantarum

ABSTRACTLactobacillus paraplantarumBGCG11, a putative probiotic strain isolated from a soft, white, artisanal cheese, produces a high-molecular-weight heteropolysaccharide, exopolysaccharide (EPS)-CG11, responsible for the ropy phenotype and immunomodulatory activity of the strain. In this study, a 26.4-kb region originating from the pCG1 plasmid, previously shown to be responsible for the production of EPS-CG11 and a ropy phenotype, was cloned, sequenced, and functionally characterized. In this region 16 putative open reading frames (ORFs), encoding enzymes for the production of EPS-CG11, were organized in specific loci involved in the biosynthesis of the repeat unit, polymerization, export, regulation, and chain length determination. Interestingly, downstream of theepsgene cluster, a putative transposase gene was identified, followed by an additionalrfbgene cluster containing therfbACBDgenes, the ones most probably responsible for dTDP-l-rhamnose biosynthesis. The functional analysis showed that the production of the high-molecular-weight fraction of EPS-CG11 was absent in two knockout mutants, one in theepsand the other in therfbgene cluster, as confirmed by size exclusion chromatography analysis. Therefore, bothepsandrfbgenes clusters are prerequisites for the production of high-molecular-weight EPS-CG11 and for the ropy phenotype of strainL. paraplantarumBGCG11.

scholarly journals Molluscicidal Polyphenols from Species of Fucaceae

2008 ◽  
Vol 3 (2) ◽  
pp. 1934578X0800300
Author(s):  
Asmita V. Patel ◽  
David C. Wright ◽  
Maricela Adrian Romero ◽  
Gerald Blunden ◽  
Michael D. Guiry
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Algal Species ◽  
Weight Fraction ◽  
The Other ◽  
Low Molecular Weight ◽  
High Molecular Weight Fraction ◽  
Bifurcaria Bifurcata ◽  
Marine Algal ◽  
Molluscicidal Activity

The aqueous fractions of the dry methanol extracts (500 ppm) of sixty marine algal species were screened for molluscicidal activity against Biomphalaria glabrata, which is an important host of the bilharzia-causing Schistosoma species. The majority of the extracts tested were inactive at the concentration used, but those of Fucus serratus, F. vesiculosus, Pelvetia canaliculata, Ascophyllum nodosum, Halidrys siliquosa, Bifurcaria bifurcata, Dictyota dichotoma and Halopithys incurva all showed significant molluscicidal activity. Treatment with polyvinylpolypyrrolidone (PVPP) either removed or drastically reduced the activity of the extracts of F. serratus, F. vesiculosus, P. canaliculata, A. nodosum, Halidrys siliquosa and Halopithys incurva, which suggested that the active compounds in the extracts of these species were polyphenolic in nature. The active extracts of the other two seaweed species did not appear to be affected by treatment with PVPP. Dialysis of the active extracts against distilled water separated them into high and low molecular weight fractions. In the case of the two Fucus species, P. canaliculata and A. nodosum, the activity resided in the high molecular weight fraction, whereas with all the other species, the activity was found in the low molecular weight fraction. 1H NMR spectroscopic examination of the active extracts confirmed that the molluscicidal components of the extracts of the Fucus species, P. canaliculata and A. nodosum were high molecular weight polyphenols.

Cartilage fucoproteins with sites for cross-linking by transglutaminase

1987 ◽  
Vol 65 (4) ◽  
pp. 280-285 ◽  
Author(s):  
J. Michael Bowness
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
High Performance ◽  
Specific Activity ◽  
Weight Fraction ◽  
Dry Weight ◽  
Insoluble Fraction ◽  
Low Molecular Weight ◽  
High Molecular Weight Fraction ◽  
Collagen Iii

Slices of various types of cartilage were incubated with either L-[6-3H]fucose or [1,4-3H(N)]putrescine. Homogenization of the slices and fractionation of the homogenates showed for both labels that an insoluble collagenase-resistant fraction had the highest specific activity (dpm/mg dry weight). Examination of an exhaustive proteolytic digest of this insoluble fraction by ion-exchange high performance liquid chromatography showed the presence of γ-glutamyl[3H]putrescine. Chromatography of solubilized [3H]fucoprotein fractions showed the presence of several low molecular weight peaks, as well as high molecular weight material. Incubation of [3H]fucoprotein extracts with transglutaminase increased the high molecular weight peaks and decreased the low molecular weight ones. Incubation of the cartilage slices with L-[3H]fucose plus 0.5 mM dansylcadaverine, an inhibitor of transglutaminase, caused a decrease in the insoluble and high molecular weight fraction relative to the low molecular weight peaks. It is hypothesized that this is due to inhibition of cross-link formation between fucoprotein components of the cartilage which are transglutaminase substrates. One major low molecular weight peak, which labels with both fucose and putrescine, corresponds in size with the 15 000 subunit of collagen III aminopropeptide, which is known to be a substrate for transglutaminase.

scholarly journals The Antiproliferative Activity of Sclerotia ofLignosus rhinocerus(Tiger Milk Mushroom)

2012 ◽  
Vol 2012 ◽  
pp. 1-5 ◽  
Author(s):  
M. L. Lee ◽  
N. H. Tan ◽  
S. Y. Fung ◽  
C. S. Tan ◽  
S. T. Ng
Keyword(s):  
Molecular Weight ◽  
Cancer Cells ◽  
High Molecular Weight ◽  
Antiproliferative Activity ◽  
Human Lung ◽  
Weight Fraction ◽  
Low Molecular Weight ◽  
Cytotoxic Action ◽  
High Molecular Weight Fraction ◽  
Human Breast

Lignosus rhinocerus, the tiger milk mushroom, is one of the most important medicinal mushrooms used by the indigenous people of Southeast Asia and China. It has been used to treat breast cancer. A cold water extract (LR-CW) prepared from the sclerotia ofL. rhinoceruscultivar was found to exhibit antiproliferative activity against human breast carcinoma (MCF-7) and human lung carcinoma (A549), withIC50of 96.7 μg/mL and 466.7 μg/mL, respectively. In comparison, LR-CW did not show significant cytotoxicity against the two corresponding human normal cells, 184B5 (human breast cell) and NL 20 (human lung cell). DNA fragmentation studies suggested that the cytotoxic action of LR-CW against cancer cells is mediated by apoptosis. Sephadex G-50 gel filtration fractionation of LR-CW yielded a high-molecular-weight and a low-molecular-weight fraction. The high-molecular-weight fraction contains mainly carbohydrate (68.7%) and small amount of protein (3.6%), whereas the low-molecular-weight fraction contains 31% carbohydrate and was devoid of protein. Only the high-molecular-weight fraction exhibited antiproliferative activity against cancer cells, withIC50of 70.0 μg/mL and 76.7 μg/mL, respectively. Thus, the cytotoxic action of the LR-CW is due to the high-molecular-weight fraction, either the proteins or protein-carbohydrate complex.

Tumor Uptake of High and Low Molecular Weight Fractions from 67Ga-Citrate Labelled Blood Plasma

1984 ◽  
Vol 23 (02) ◽  
pp. 59-61 ◽  
Author(s):  
M. C. Crone ◽  
P. Thouvenot ◽  
F. Brunotte ◽  
C. Marchai ◽  
J. Robert ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Blood Plasma ◽  
High Molecular Weight ◽  
Gel Filtration ◽  
Weight Fraction ◽  
Blood Protein ◽  
Low Molecular Weight ◽  
High Molecular Weight Fraction ◽  
67Ga Citrate

SummaryBlood plasma from tumor-bearing rats was incubated with 67Ga-citrate, and two fractions of high molecular weight (proteins) and low molecular weight were isolated by dialysis and by gel-filtration chromatography. Both fractions showed a different in vivo uptake by DS-sarcoma-bearing animals, the high molecular weight fraction being accumulated to a lesser extent. Compared to 67Ga-citrate the low molecular weight fraction showed a different uptake which for most tissues was significatively higher. This behavior suggests the presence of 67Ga in chemical forms other than citrate in the low molecular weight fraction. The lower uptake of the blood protein fraction is discussed.

STRUCTURE AND CHEMICAL CONTENTS OF MUCILAGE CELLS IN PSEUDOLARIX AND ABIES BARK

IAWA Journal ◽  
2015 ◽  
Vol 36 (3) ◽  
pp. 300-310
Author(s):  
Fengping Jia ◽  
Jiayan Luo ◽  
Ryosuke Takahashi ◽  
Chie Uehira ◽  
Shinichi Kitamura ◽  
...  
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Average Length ◽  
Weight Fraction ◽  
Ruthenium Red ◽  
Low Molecular Weight ◽  
High Molecular Weight Fraction ◽  
H Nmr ◽  
Molecular Weight Fractions ◽  
Chemical Contents

Mucilage is extruded from the bark of Pseudolarix amabilis and Abies nephrolepis upon injury. The aim of this study was to characterize the structure and chemical contents of mucilage extruded from mucilage cells (MCs) in the bark of these species. A large number of MCs containing translucent or dark materials in their lumina were observed in the secondary phloem of P. amabilis and A. nephrolepis. The translucent or dark materials in MCs stained positive with ruthenium red and PAS, indicating the presence of polysaccharides. The average length and diameter of MC in P. amabilis were 1500 μm and 254 μm, respectively, and the corresponding values for A. nephrolepis were 419 μm and 166 μm. Chemical analysis of low molecular weight fractions prepared from mucilage by HPAEC-PAD showed sucrose, glucose and fructose peaks, and in addition galacturonic acid and fucose peaks. Furthermore, 1H NMR spectra for the high molecular weight fraction showed the signals characteristic of pectin. This demonstrates that the mucilage consists mainly of low molecular weight carbohydrates and high molecular weight polysaccharide pectin.

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