scholarly journals Polyamine degradation in foetal and adult bovine serum

1982 ◽  
Vol 202 (3) ◽  
pp. 603-611 ◽  
Author(s):  
W A Gahl ◽  
H C Pitot
Keyword(s):  
Bovine Serum ◽  
Oxidase Activity ◽  
Degrading Enzyme ◽  
Substrate Preferences ◽  
Adult Bovine Serum ◽  
Putrescine Oxidase ◽  
20 Nm ◽  
Spermidine Oxidase ◽  
Foetal Bovine Serum

1. Using protein-separative chromatographic procedures and assays specific for putrescine oxidase and spermidine oxidase, adult bovine serum was found to contain a single polyamine-degrading enzyme with substrate preferences for spermidine and spermine. Apparent Km values for these substrates were approx. 40 microM. The apparent Km for putrescine was 2 mM. With spermidine as substrate, the Ki values for aminoguanidine (AM) and methylglyoxal bis(guanylhydrazone) (MGBG) were 70 microM and 20 microM respectively. 2. Bovine serum spermidine oxidase degraded spermine to spermidine to putrescine and N8-acetylspermidine to N-acetylputrescine. Acrolein was produced in all these reactions and recovered in quantities equivalent to H2O2 recovery. 3. Spermidine oxidase activity was present in foetal bovine serum, but increased markedly after birth to levels in adult serum that were almost 100 times the activity in foetal bovine serum. 4. Putrescine oxidase, shown to be a separate enzyme from bovine serum spermidine oxidase, was present in foetal bovine serum but absent from bovine serum after birth. This enzyme displayed an apparent Km for putrescine of 2.6 microM. The enzyme was inhibited by AM and MGBG with Ki values of 20 nM. Putrescine, cadaverine and 1,3-diaminopropane proved excellent substrates for the enzyme compared with spermidine and spermine, and N-acetylputrescine was a superior substrate to N1- or N8-acetylspermidine.

scholarly journals Separation of putrescine oxidase and spermidine oxidase in foetal bovine serum with the aid of a specific radioactive assay of spermidine oxidase

1980 ◽  
Vol 187 (1) ◽  
pp. 197-204 ◽  
Author(s):  
W A Gahl ◽  
A M Vale ◽  
H C Pitot
Keyword(s):  
Bovine Serum ◽  
Oxidase Activity ◽  
Gel Filtration ◽  
Major Product ◽  
Sephadex Column ◽  
Exchange Column ◽  
Cation Exchange Column ◽  
Putrescine Oxidase ◽  
Spermidine Oxidase ◽  
Foetal Bovine Serum

1. A sensitive and specific assay for spermidine oxidase is described. The method involves the separation of [14C]spermidine (substrate) from [14C]putrescine (product) and other 14C-labelled products on a Dowex 50 cation-exchange column: 92% of the putrescine applied to the column was eluted by 2.3 M-HCl, but this treatment left 96% of the spermidine bound to the column. Unchanged spermidine could be removed from the column by elution with 6 M-HCl. 2. By means of this assay, foetal and adult bovine serum were each shown to contain spermidine oxidase activity, putrescine being a major product of the oxidation of spermidine by the serum enzymes. 3. In foetal bovine serum, spermidine oxidase activity is separable from putrescine oxidase activity by chromatography on a cadaverine-Sephadex column, by gel filtration and by ion-exchange column chromatography. Putrescine oxidase was purified 1900-fold and spermidine oxidase 130-fold by these procedures. The former oxidized putrescine but not spermidine, and spermidine oxidase exhibited no activity with putrescine as substrate.

scholarly journals Foetal bovine serum influence on in vitro extracellular vesicle analyses

2021 ◽  
Vol 10 (3) ◽  
Author(s):  
Brandon M. Lehrich ◽  
Yaxuan Liang ◽  
Massimo S. Fiandaca
Keyword(s):  
Bovine Serum ◽  
Extracellular Vesicle ◽  
Foetal Bovine Serum

scholarly journals Optimisation of the lymphocyte proliferation assay in rainbow trout (Oncorhynchus mykiss)

2019 ◽  
Vol 64 (No. 12) ◽  
pp. 547-557
Author(s):  
H Minarova ◽  
M Palikova ◽  
J Mares ◽  
E Syrova ◽  
J Blahova ◽  
...  
Keyword(s):  
Rainbow Trout ◽  
Oncorhynchus Mykiss ◽  
Bovine Serum ◽  
Lymphocyte Proliferation ◽  
Head Kidney ◽  
Pokeweed Mitogen ◽  
Lymphocyte Proliferation Assay ◽  
Proliferation Assay ◽  
Rainbow Trout Oncorhynchus Mykiss ◽  
Foetal Bovine Serum

The lymphocyte proliferation assay is a valuable method used for the evaluation of the fish immune system. However, there are many variations and optimal results are not always obtained. Unification is necessary to ensure the comparability between different studies. The aim of this study was to optimise the lymphocyte proliferation assay in rainbow trout (Oncorhynchus mykiss). This goal included the determination of the optimal incubation length, serum type, incubation temperature, type of mitogen and its concentration, and anticoagulant. The peripheral blood and head kidney lymphocytes were isolated by density gradient centrifugation. Subsequently, the cells were incubated for 3–8 days with different mitogens (pokeweed mitogen 5, 10 and 50 µg/ml, concanavalin A 1, 10 and 20 µg/ml, phytohaemagglutinin 25, 50 and 100 µg/ml, lipopolysaccharide 1, 50 and 100 µg/ml). The use of the different serum types (foetal bovine serum, trout serum), incubation temperatures (10–20 °C) and anticoagulants (heparin, EDTA) was compared. Labelled thymidine was used to evaluate the assay. The best results were obtained after seven days of incubation at 15 °C with foetal bovine serum (FBS). The head kidney lymphocytes showed the highest proliferative response with 50 µg/ml phytohaemagglutinin. With the peripheral blood lymphocytes (heparin and EDTA), the best results were obtained with 50 µg/ml pokeweed mitogen. The highest proliferation levels were detected with heparinised blood. In conclusion, optimisation of this assay contributes to the improved assessment of the rainbow trout immune function.

scholarly journals ln vitro isolation of Ehrlichia ruminantium from ovine blood into lxodes scapularis (lDE8) cell cultures

2008 ◽  
Vol 75 (2) ◽  
Author(s):  
E. Zweygarth ◽  
A. I. Josemans ◽  
H. C. Steyn
Keyword(s):  
Endothelial Cells ◽  
Cell Cultures ◽  
Causative Agent ◽  
Bovine Serum ◽  
Infected Sheep ◽  
Ehrlichia Ruminantium ◽  
Domestic Ruminants ◽  
Bovine Endothelial Cells ◽  
Foetal Bovine Serum

Four stocks of Ehrlichiar uminantium (Welgevonden, Ball3, Nonile and Blaauwkrans), the causative agent of heartwater in domestic ruminants, were isolated into lxodes capularis (lDE8) tick cells using the leukocyte fraction of the blood of infected sheep. Organisms of two of the E. ruminantrum stocks (Welgevonden and Blaauwkrans) propagated in IDEB cells were also successfully used to infect bovine endothelial cells. All stocks were successfully propagated in IDEB cells using Dulbecco's modified Eagle's medium nutrient mixture Ham F-12c ontaining 10% foetal bovine serum (FBS). The technique should be included in any attempt to isolate uncharacterized E. ruminantium stocks.

Fretting corrosion behaviour of Ti-6Al-4V reinforced with zirconia in foetal bovine serum

2019 ◽  
Vol 100 ◽  
pp. 103392 ◽  
Author(s):  
Lerato Semetse ◽  
Babatunde Abiodun Obadele ◽  
Lerato Raganya ◽  
Jean Geringer ◽  
Peter Apata Olubambi
Keyword(s):  
Bovine Serum ◽  
Corrosion Behaviour ◽  
Fretting Corrosion ◽  
Foetal Bovine Serum

scholarly journals UDP-sugar metabolism in Swarm rat chondrosarcoma chondrocytes

1993 ◽  
Vol 290 (2) ◽  
pp. 563-570 ◽  
Author(s):  
C Sweeney ◽  
D Mackintosh ◽  
R M Mason
Keyword(s):  
Anion Exchange ◽  
Cell Cultures ◽  
Half Life ◽  
Bovine Serum ◽  
Adenine Nucleotides ◽  
Specific Radioactivity ◽  
Sugar Metabolism ◽  
Amino Sugar ◽  
Foetal Bovine Serum ◽  
Pool Sizes

UDP-sugars and adenine nucleotides were extracted from freshly isolated chondrocytes and primary cell cultures and analysed by anion-exchange h.p.l.c. The pool sizes of UDP-N-acetylglucosamine, UDP-N-acetylgalactosamine, UDP-glucose-galactose, UDP-glucuronate and UDP-xylose were 2.9, 1.2, 2.5, 0.6 and 0.03 nmol/10(6) freshly isolated chondrocytes. When chondrocytes were maintained in Dulbecco's modified Eagle medium supplemented with 15% foetal-bovine serum, synthesis of [35S]proteoglycan and [3H]protein decreased over the first 48 h in culture, as did the pools of UDP-glucuronate and ATP. In contrast, the size of the UDP-N-acetylhexosamine pools underwent little change during culture. [35S]Proteoglycan and [3H]protein syntheses were stimulated in cultures supplemented with serum or insulin compared with those maintained in medium alone, in agreement with previous results. However, the UDP-sugar pool sizes were the same in both supplemented and non-supplemented cultures. In cultures maintained in the presence of [1-3H]glucose, the UDP-sugars were labelled to a constant 3H specific radioactivity which was very similar to that of the labelling medium. UDP-N-acetylhexosamines were labelled to constant 3H specific radioactivity with [6-3H]glucosamine as a precursor, but only about 1 in 375 of these UDP-sugars was derived from the amino sugar in the presence of glucose. The half-life (t1/2) for UDP-hexoses, UDP-glucuronate and UDP-N-acetylhexosamines was about 12, 12 and 50 min respectively.

Effect of additives in medium on in-vitro maturation of goat oocytes

2019 ◽  
Author(s):  
D. Borah ◽  
R.K. Biswas
Keyword(s):  
Follicular Fluid ◽  
Bovine Serum ◽  
In Vitro Maturation ◽  
Cumulus Cells ◽  
Sodium Pyruvate ◽  
Nuclear Maturation ◽  
Effect Of Additives ◽  
Additive Control ◽  
Foetal Bovine Serum

Present study was carried out to find the effect of combining EGF with IGF, cysteine and sodium pyruvate singly as additive in a medium consisting of TCM-199 + 100 µl/ml foetal bovine serum + 100 µM/ml cysteamine + 1 µg/ml 17â- Oestradiol + 5 µg/ml pFSH + 5µg/ml oLH + 10 per cent follicular fluid and 10 per cent oestrous goat serum on in-vitro maturation (IVM) of caprine oocytes on incubation at 38.50C for 24 hours in a CO2 incubator maintaining 5 per cent CO2 under humidified condition. The additives comprised 10 ng/ml EGF + 50 ng/ml IGF-1, 10 ng/ml EGF + 600 µM/ml cysteine and 10 ng/ ml EGF + 0.2 mM/ml sodium pyruvate. The IVM rate of oocytes on the basis of cumulus cells expansion and nuclear maturation was found to be significantly higher (P less than 0.05) with EGF + IGF-1 (88.74 ± 1.85% and 61.71 ± 1.61%) than with EGF + sodium pyruvate (82.86 ± 0.97% and 54.62 ± 1.88%), EGF + cysteine ( 78.58 ± 1.45% and 49.02 ± 1.52%) and without additive (control) (75.27 ± 1.58% and 43.03 ± 1.48%).

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