Rapid assay of poly(ADP-ribose) glycohydrolase

1987 ◽  
Vol 65 (7) ◽  
pp. 668-673 ◽  
Author(s):  
Luc Ménard ◽  
Guy G. Poirier

We have developed a rapid, highly reproducible assay to determine poly(ADP-ribose) glycohydrolase activity which measures directly the appearance of the reaction product. We also analysed the majority of different techniques which are used to determine poly(ADP-ribose) glycohydrolase activity and found that the apparent activity can vary extensively depending on the method used. Thin-layer chromatography using PEI-F–cellulose was the only method which evaluated directly the specific release of ADP-ribose; by comparison with this method, the other procedures gave an over- or under-estimation of 2- to 10-fold of the enzymatic activity. A rapid method of affinity chromatography has also been developed to synthesize and purify in high yield poly(ADP-ribose) (35% conversion of 1 mM NAD to poly (ADP-ribose)).

1997 ◽  
Vol 75 (7) ◽  
pp. 1066-1072 ◽  
Author(s):  
Steven G. Burgin ◽  
Fiona F. Hunter

Adult black flies were sampled by sweep-netting vegetation in four habitats within Algonquin Provincial Park, Ontario: Davies Bog, the airfield, deciduous habitat, and coniferous habitat. Sugars in the crops and midguts of female flies (n = 773) were tested by thin-layer chromatography to determine whether the flies had fed on nectar or homopteran honeydew. Melezitose and stachyose were used as honeydew-indicator sugars. For Simulium venustum, it was found that significantly fewer black flies (19%) from the airfield contained honeydew sugars than black flies from the other three sites (34% from Davies Bog; 36% from deciduous habitat; 25% from coniferous habitat). We argue that black flies will feed on nectar or honeydew according to availability.


Endocrinology ◽  
2003 ◽  
Vol 144 (2) ◽  
pp. 575-580 ◽  
Author(s):  
Jean D. Wilson ◽  
Richard J. Auchus ◽  
Michael W. Leihy ◽  
Oleg L. Guryev ◽  
Ronald W. Estabrook ◽  
...  

The synthetic pathway by which 5α-androstane-3α,17β-diol (5α-adiol) is formed in the testes of tammar wallaby pouch young was investigated by incubating testes from d 20–40 males with various radioactive precursors and analyzing the metabolites by thin-layer chromatography and HPLC. [3H]Progesterone was converted to 17-hydroxyprogesterone, which was converted to 5α-adiol by two pathways: One involves the formation of testosterone and dihydrotestosterone as intermediates, and the other involves formation of 5α-pregnane-3α,17α-diol-20-one (5α-pdiol) and androsterone as intermediates. Formation of 5α-adiol from both [3H]testosterone and [3H]progesterone was blocked by the 5α-reductase inhibitor 4MA. The addition of nonradioactive 5α-pdiol blocked the conversion of [3H]progesterone to 5α-adiol, and [3H]5α-pdiol was efficiently converted to androsterone and 5α-adiol. We conclude that expression of steroid 5α-reductase in the developing wallaby testes allows formation of 5α-reduced androgens by a pathway that does not involve testosterone as an intermediate.


1999 ◽  
Vol 269 (1) ◽  
pp. 17-20 ◽  
Author(s):  
Vijaya B. Kumar ◽  
Adonis E. Bernardo ◽  
M.M. Alshaher ◽  
M. Buddhiraju ◽  
R. Purushothaman ◽  
...  

1973 ◽  
Vol 30 (03) ◽  
pp. 557-566 ◽  
Author(s):  
S Renaud ◽  
P Gautheron ◽  
H Rosenstein

SummaryPlatelets collected with an EDTA solution and simply washed in an incomplete Tyrode’s presented clotting times in the recalcification (man and rat) and the Stypven (rat) tests that were practically identical to those of the PRP when slow speed centrifugation was used (800 G in man, 1000 G in rat). This was demonstrated, in 6 pools of 5 rats each and in 6 men, by comparing the clotting activity of the citrated platelet-rich plasma to that of the platelets washed and resuspended in the citrated platelet-poor plasma, for platelet counts ranging from 1 × 105 to 10 × 105/mm3. In contrast, centrifugation of platelets at 3000 G markedly affected these clotting activities, as was shown in an additional study comprising 6 pools of 3 rats.Finally, the clotting activity of platelets totally disrupted by sonication appears to be identical quantitatively in both man and rats to that of the total phospholipids extracted from these platelets and separated from the other lipids by thin-layer chromatography and resuspended in plasma by sonication.


1966 ◽  
Vol 49 (2) ◽  
pp. 468-472
Author(s):  
Arthur W Davidson

Abstract Twelve collaborators from six laboratories used gas-liquid and thin layer chromatography to measure BHC, p,p′-DDT, and endrin residues on apricots and strawberries. Recoveries for both crops by gas chromatography averaged: BHC, 104 ± 23 %; p,p′-DDT, 102 ± 10%; endrin, 96 ±14%. Recoveries for both crops by thin layer chromatography averaged: BHC, 98 ± 29%; p,p′-DDT, 97 ± 18%; and endrin, 96 ± 16%.


1970 ◽  
Vol 48 (1) ◽  
pp. 49-53 ◽  
Author(s):  
Dianne Fahselt

The main anthocyanin pigments of the genus Dicentra have been characterized through the use of spectroscopy and chromatography. This combination of techniques, particularly suited to anthocyanins, has demonstrated that all are glycosides of cyanidin (3,5,7,3′,4′-pentahydroxyflavylium) and that none are acylated. The simplest, found in several species, is cyanidin-3-monoglucoside, while the most complex is cyanidin-3-rhamnoglucosido-7-xyloside. This triglycoside is present in only one species as well as in certain experimental interspecific hybrids, and this is its first reported occurrence in nature. A rapid method is described for identification of anthocyanin sugars using thin-layer chromatography.


Sign in / Sign up

Export Citation Format

Share Document