scholarly journals Degradation of nucleic acids in the rumen

1973 ◽  
Vol 29 (2) ◽  
pp. 331-345 ◽  
Author(s):  
A. B. McAlian ◽  
R. H. Smith
Keyword(s):  
Nucleic Acids ◽  
Thin Layer ◽  
Degradation Products ◽  
Rumen Fluid ◽  
The Other ◽  
Pyrimidine Bases ◽  
Layer Chromatography ◽  
Rna And Dna

1. Nucleic acids introduced into the rumens of calves, or incubated with calf, sheep or cow rumen contents in vitro, were rapidly destroyed.2. The degradation products formed were separated and identified by means of column chromatography on Sephadex G-10 Dextran gel and thin-layer chromatography on cellulose.3. In vitro, RNA was rapidly (within 1 h) converted into ultrafilterable oligo-and mono-nucleotides, nucleosides, purine and pyrimidine bases. After 4 h, only the bases xanthine, hypoxanthine and uracil remained, having increased at the expense of the other constituents.4. DNA gave similar products but with a much greater proportion of ultrafilterable oligoand mono-nucleotide material which remained as a major component even after 4 h. The only bases present in appreciable amounts were thymine, hypoxanthine, uracil and xanthine.5. The same products accumulated temporarily in vivo, after addition of RNA or DNA to the rumens of calves, and were found also, in small amounts, in corresponding samples of duodenal digesta. The products disappeared from the rumen at a greater rate than could be accounted for by transfer to the duodenum.6. Cell-free preparations from calf rumen fluid contained enzymes which converted RNA and DNA into products which appeared to be ultrafilterable oligonucleotides.7. When ground hay was incubated with whole rumen contents the nucleic acids in the hay were degraded to a mixture of nucleotides, nucleosides and bases, almost as readily as were pure nucleic acids.

Phytochemical, Analytical and Medicinal Studies of Holoptelea integrifolia Roxb. Planch - A Review

2019 ◽  
Vol 5 (4) ◽  
pp. 270-277 ◽  
Author(s):  
Vijay Kumar ◽  
Simranjeet Singh ◽  
Ragini Bhadouria ◽  
Ravindra Singh ◽  
Om Prakash
Keyword(s):  
Liquid Chromatography ◽  
Thin Layer ◽  
Thin Layer Chromatography ◽  
High Performance ◽  
Biologically Active ◽  
Toxicological Studies ◽  
Wide Range ◽  
Layer Chromatography

Holoptelea integrifolia Roxb. Planch (HI) has been used to treat various ailments including obesity, osteoarthritis, arthritis, inflammation, anemia, diabetes etc. To review the major phytochemicals and medicinal properties of HI, exhaustive bibliographic research was designed by means of various scientific search engines and databases. Only 12 phytochemicals have been reported including biologically active compounds like betulin, betulinic acid, epifriedlin, octacosanol, Friedlin, Holoptelin-A and Holoptelin-B. Analytical methods including the Thin Layer Chromatography (TLC), High-Performance Thin Layer Chromatography (HPTLC), High-Performance Liquid Chromatography (HPLC) and Liquid Chromatography With Mass Spectral (LC-MS) analysis have been used to analyze the HI. From medicinal potency point of view, these phytochemicals have a wide range of pharmacological activities such as antioxidant, antibacterial, anti-inflammatory, and anti-tumor. In the current review, it has been noticed that the mechanism of action of HI with biomolecules has not been fully explored. Pharmacology and toxicological studies are very few. This seems a huge literature gap to be fulfilled through the detailed in-vivo and in-vitro studies.

scholarly journals EXTRACTION, ISOLATION AND STRUCTURAL ELUCIDATION OF FLAVONOID FROM CHROZOPHORA PLICATA LEAVES AND EVALUATION OF ITS ANTIOXIDATIVE POTENTIALS

2017 ◽  
Vol 10 (4) ◽  
pp. 460
Author(s):  
Kadiri Sunil Kumar ◽  
Avanapu Srinivasa Rao
Keyword(s):  
Thin Layer ◽  
Superoxide Anion ◽  
Methanolic Extract ◽  
Reducing Power ◽  
Reducing Power Assay ◽  
Isolated Compound ◽  
Layer Chromatography ◽  
Column Chromatographic

Objective: This investigation involves the extraction, isolation, and characterization of flavonoid from a Euphorbiaceae family plant Chrozophoraplicata followed by evaluation of its antioxidant principles.Methods: The dried leaves were subjected to sequential soxhlation with polar and nonpolar solvents. Methanolic extract reveals the presence of largeamount of flavonoids. Methanolic extract was subjected to isolation using column chromatographic analysis with solvents such as petroleum ether,chloroform, hexane, ethyl acetate, methanol, and water. Further, the isolated compound was subjected to thin layer chromatography technique andspectral analysis such as infrared, 1HNMR, 13CNMR, mass spectroscopy, and high performance thin layer chromatography (HPTLC) finger printingtechniques. The compound was evaluated for in vitro antioxidant studies using 2,2-diphenyl-1-picrylhydrazyl (DPPH), NO assay, reducing power assay,H2O2 scavenging assay, superoxide anion scavenging assay and β-Carotene linoleate system and in vivo antioxidative studies using carbon tetrachloride(CCl4), and acetaminophen intoxicated rats.Results: The compound was isolated in methanol:water in the ratio of 80:20 using column chromatographic technique. On the basis of phytochemical,chromatographic, and spectral analysis, the isolated compound was identified as kaempferol and finally with HPTLC finger printing technique it wasfound that the Rf value of the isolated compound was found to be 0.58 which is nearly similar to the Rf value of standard kaempferol (0.55). Hence,the isolated compound was confirmed as kaempferol and is structurally elucidated as 3,5,7-trihydroxy-2-(4-hydroxyphenyl)chromen-4-one. Thiscompound was isolated for the first time from the C. plicata leaves. The in vitro antioxidant assay of isolated flavonoid has shown a dose-dependentincrease in free radical scavenging activity using DPPH, no assay, reducing power assay, H2O2 scavenging assay, superoxide anion scavenging assay, andβ-carotene linoleate system. Further, the methanolic extract of C. plicata (MECP) leaves was subjected to single dose acute toxicity study for 14 days infemale rats on the basis of OECD guidelines 423 and the therapeutically selected doses were 200 mg/kg and 400 mg/kg. In vivo antioxidant studies inCCl4 and acetaminophen intoxicated rats indicated that the MECP leaves have significantly decreased lipid peroxidation in a dose-dependent mannerand increased the levels of catalase, superoxide dismutase, and glutathione.Conclusions: By the above results, it was concluded that the isolated compound from C. plicata leaves was confirmed as kaempferol and it possessessignificant antioxidative potentials.Keywords: Chrozophora plicata leaves, Flavonoids, Extraction, Isolation, Characterization, Methanolic extract, Antioxidant activity, Carbontetrachloride, Acetaminophen.

scholarly journals Metabolism of Zearalenone in the Rumen of Dairy Cows with and without Application of a Zearalenone-Degrading Enzyme

Toxins ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 84
Author(s):  
Christiane Gruber-Dorninger ◽  
Johannes Faas ◽  
Barbara Doupovec ◽  
Markus Aleschko ◽  
Christian Stoiber ◽  
...  
Keyword(s):  
Dairy Cows ◽  
Animal Feed ◽  
Degradation Products ◽  
Rumen Fluid ◽  
Feed Additive ◽  
Estrogenic Effects ◽  
Low Concentrations ◽  
Holstein Friesian

The mycotoxin zearalenone (ZEN) is a frequent contaminant of animal feed and is well known for its estrogenic effects in animals. Cattle are considered less sensitive to ZEN than pigs. However, ZEN has previously been shown to be converted to the highly estrogenic metabolite α-zearalenol (α-ZEL) in rumen fluid in vitro. Here, we investigate the metabolism of ZEN in the reticulorumen of dairy cows. To this end, rumen-fistulated non-lactating Holstein Friesian cows (n = 4) received a one-time oral dose of ZEN (5 mg ZEN in 500 g concentrate feed) and the concentrations of ZEN and ZEN metabolites were measured in free rumen liquid from three reticulorumen locations (reticulum, ventral sac and dorsal mat layer) during a 34-h period. In all three locations, α-ZEL was the predominant ZEN metabolite and β-zearalenol (β-ZEL) was detected in lower concentrations. ZEN, α-ZEL and β-ZEL were eliminated from the ventral sac and reticulum within 34 h, yet low concentrations of ZEN and α-ZEL were still detected in the dorsal mat 34 h after ZEN administration. In a second step, we investigated the efficacy of the enzyme zearalenone hydrolase ZenA (EC 3.1.1.-, commercial name ZENzyme®, BIOMIN Holding GmbH, Getzersdorf, Austria) to degrade ZEN to the non-estrogenic metabolite hydrolyzed zearalenone (HZEN) in the reticulorumen in vitro and in vivo. ZenA showed a high ZEN-degrading activity in rumen fluid in vitro. When ZenA was added to ZEN-contaminated concentrate fed to rumen-fistulated cows (n = 4), concentrations of ZEN, α-ZEL and β-ZEL were significantly reduced in all three reticulorumen compartments compared to administration of ZEN-contaminated concentrate without ZenA. Upon ZenA administration, degradation products HZEN and decarboxylated HZEN were detected in the reticulorumen. In conclusion, endogenous metabolization of ZEN in the reticulorumen increases its estrogenic potency due to the formation of α-ZEL. Our results suggest that application of zearalenone hydrolase ZenA as a feed additive may be a promising strategy to counteract estrogenic effects of ZEN in cattle.

SULFATIDES AND GLYCOLIPIDS IN PLATELETS AND ENDOTHELIAL CELLS

1987 ◽  
Author(s):  
K P Schick ◽  
S Shapiro ◽  
G Tuszynski ◽  
J Slawek
Keyword(s):  
Endothelial Cells ◽  
Thin Layer ◽  
Thin Layer Chromatography ◽  
Specific Binding ◽  
Blood Platelets ◽  
Primary Cultures ◽  
Galactosyl Ceramide ◽  
Layer Chromatography

Sulfatides are sulfated glycolipids which are negatively charged and thought to influence receptor mediated activities. Sulfatides have the capacity to provide a surface for the initiation of in vitro coagulation tests and these acidic lipids represent the potential biological surface for the initiation of the contact and intrinsic systems in vivo. Several sulfatides have been demonstrated in blood platelets. We have investigated sulfatides and other glycolipids in endothelial cells and platelets in order to define the cellular sources for sulfatides that would be available for influencing hemostasis. Endothelial cells were derived from primary cultures of human umbilical veins and human platelets were obtained from freshly-collected blood. Cellular lipids were extracted by the Folch method. Sulfatides and glycolipids were purified by silicic acid chromatography, separated by thin-layer chromatography, and quantitated by the assay of sphingosine. Glycolipids were also analyzed by HPLC. Globoside was found to be the predominant glycolipid in endothelial cells while lactosyl ceramide was the predominant glyco-lipid in platelets. Sulfatides were detected by two approaches: 1) Sulfatide synthesis by the incorporation of [35S]-Sulfate; 2) The specific binding of [125I]-thrombospondin and [125I]-von Willebrand’s factor (vWF) to sulfatides separated by thin-layer chromatography (TLC). Several sulfatides were identified in endothelial cells and platelets by virtue of the incorporation of [35S]-sulfate into glycolipids separated by TLC. [125I]-TSP and [125I]-vWF bound to the glycolipids that had incorporated [35S]-sulfate. [35S]-sulfate was primarily incorporated into sulfated galactosyl ceramide but both cells also synthesized complex glycolipids. TSP and vWF were shown to bind to sulfated galactosyl ceramide, a band that comigrated with glycosyl ceramide as well as with two more complex sulfatides in both cells. However, differences in sulfatide synthesis and binding of TSP to sulfatides were observed in endothelial cells from that in platelets. The study indicates that endothelial cells and platelets contain several sulfatides and thus are potential sources for sulfatides for the initiation of coagulation.

EXTRACTION AND CHEMICAL CHARACTERISTICS OF ANOREXIGENIC AND FAT-MOBILIZING SUBSTANCES FROM RAT URINE

1964 ◽  
Vol 42 (5) ◽  
pp. 647-655 ◽  
Author(s):  
John R. Beaton ◽  
A. J. Szlavko ◽  
J. A. F. Stevenson
Keyword(s):  
Amino Acids ◽  
Thin Layer ◽  
Glutamic Acid ◽  
Thin Layer Chromatography ◽  
Alkaline Solution ◽  
Chemical Characteristics ◽  
Rat Urine ◽  
Layer Chromatography

An anorexigenic and fat-mobilizing substance (FMS I), extracted from the urine of fasting rats, has been further fractionated into two materials, FMS IA and FMS IB, soluble in alkaline solution or water respectively. These two fractions have been shown to be chemically distinct by thin layer chromatography, electrophoresis, and analyses for nitrogen, carbohydrate, hexosamine, phosphorus, and "cholesterol". Further, the lipolytic activities of these extracts in vitro differ and are in the order FMS IB > FMS I > FMS IA. It has been tentatively concluded that FMS I and IA contain the 17 amino acids tryptophane, phenylalanine, leucine, arginine, isoleucine, tyrosine, valine, alanine, proline, serine, histidine, glycine, aspartic acid, glutamic acid, cystine, threonine, and lysine. FMS IB appears to contain the same amino acids with the exception of valine (which is absent). These are complex substances, the precise nature of which remains to be elucidated. It appears that the anorexigenic property of FMS I is attributable to the IA component, and the fat-mobilizing property in vivo to the IB component.

Enhancement of ADP-induced Platelet Aggregation by Adrenaline in Vivo and Its Prevention

1973 ◽  
Vol 29 (02) ◽  
pp. 490-498 ◽  
Author(s):  
Hiroh Yamazaki ◽  
Itsuro Kobayashi ◽  
Tadahiro Sano ◽  
Takio Shimamoto
Keyword(s):  
Platelet Count ◽  
Platelet Aggregation ◽  
Platelet Rich Plasma ◽  
The Other ◽  
Endothelial Surface ◽  
Important Interaction ◽  
Blood Coagulability ◽  
The Difference

SummaryThe authors previously reported a transient decrease in adhesive platelet count and an enhancement of blood coagulability after administration of a small amount of adrenaline (0.1-1 µg per Kg, i. v.) in man and rabbit. In such circumstances, the sensitivity of platelets to aggregation induced by ADP was studied by an optical density method. Five minutes after i. v. injection of 1 µg per Kg of adrenaline in 10 rabbits, intensity of platelet aggregation increased to 115.1 ± 4.9% (mean ± S. E.) by 10∼5 molar, 121.8 ± 7.8% by 3 × 10-6 molar and 129.4 ± 12.8% of the value before the injection by 10”6 molar ADP. The difference was statistically significant (P<0.01-0.05). The above change was not observed in each group of rabbits injected with saline, 1 µg per Kg of 1-noradrenaline or 0.1 and 10 µg per Kg of adrenaline. Also, it was prevented by oral administration of 10 mg per Kg of phenoxybenzamine or propranolol or aspirin or pyridinolcarbamate 3 hours before the challenge. On the other hand, the enhancement of ADP-induced platelet aggregation was not observed in vitro, when 10-5 or 3 × 10-6 molar and 129.4 ± 12.8% of the value before 10∼6 molar ADP was added to citrated platelet rich plasma (CPRP) of rabbit after incubation at 37°C for 30 second with 0.01, 0.1, 1, 10 or 100 µg per ml of adrenaline or noradrenaline. These results suggest an important interaction between endothelial surface and platelets in connection with the enhancement of ADP-induced platelet aggregation by adrenaline in vivo.

Comparison of High Purity Factor IX Concentrates and a Prothrombin Complex Concentrate in a Canine Model of Thrombogenicity

1991 ◽  
Vol 66 (05) ◽  
pp. 609-613 ◽  
Author(s):  
I R MacGregor ◽  
J M Ferguson ◽  
L F McLaughlin ◽  
T Burnouf ◽  
C V Prowse
Keyword(s):  
High Purity ◽  
Time Course ◽  
Prothrombin Complex Concentrate ◽  
Degradation Products ◽  
Canine Model ◽  
Factor Ix ◽  
In Vitro Tests ◽  
Albumin Infusion

SummaryA non-stasis canine model of thrombogenicity has been used to evaluate batches of high purity factor IX concentrates from 4 manufacturers and a conventional prothrombin complex concentrate (PCC). Platelets, activated partial thromboplastin time (APTT), fibrinogen, fibrin(ogen) degradation products and fibrinopeptide A (FPA) were monitored before and after infusion of concentrate. Changes in FPA were found to be the most sensitive and reproducible indicator of thrombogenicity after infusion of batches of the PCC at doses of between 60 and 180 IU/kg, with a dose related delayed increase in FPA occurring. Total FPA generated after 100-120 IU/kg of 3 batches of PCC over the 3 h time course was 9-12 times that generated after albumin infusion. In contrast the amounts of FPA generated after 200 IU/kg of the 4 high purity factor IX products were in all cases similar to albumin infusion. It was noted that some batches of high purity concentrates had short NAPTTs indicating that current in vitro tests for potential thrombogenicity may be misleading in predicting the effects of these concentrates in vivo.

Accumulation of Macromolecular Particles in the Reticuloendothelial System (RES) Mediated by Platelet Aggregation and Disaggregation

1969 ◽  
Vol 22 (03) ◽  
pp. 496-507 ◽  
Author(s):  
W.G van Aken ◽  
J Vreeken
Keyword(s):  
Platelet Aggregation ◽  
Degradation Products ◽  
Reticuloendothelial System ◽  
Caproic Acid ◽  
Carbon Particles ◽  
Carbon Clearance ◽  
Aggregation And Disaggregation ◽  
Platelet Aggregates

SummaryCarbon particles cause platelet aggregation in vitro and in vivo. Prior studies established that substances which modify thrombocyte aggregation also influence the rate at which carbon is cleared from the blood.This study was performed in order to elucidate the mechanism by which the carbon-platelet aggregates specifically accumulate in the RES.Activation of fibrinolysis by urokinase or streptokinase reduced the carbon clearance rate, probably due to generated fibrinogen degradation products (FDP). Isolated FDP decreased the carbon clearance and caused disaggregation of platelets and particles in vitro. Inhibition of fibrinolysis by epsilon-amino-caproic acid (EACA), initially accelerated the disappearance of carbon and caused particle accumulation outside the RES, predominantly in the lungs. It is supposed that platelet aggregation and locally activated fibrinolysis act together in the clearance of particles. In the normal situation the RES with its well known low fibrinolytic activity, becomes the receptor of the particles.

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