Putrescine determination

AdoMet is a critical branch point metabolite. In its decarboxylated form, it serves as aminopropyl group donor in polyamine synthesis as AdoMet it is the methyl group donor for most transmethylation reactions (Fig. 7.1). In eukaryotic cells AdoMet decarboxylase is activated by putrescine and is the rate-limiting step of polyamine biosynthesis. Since production of dc-AdoMet is irreversible (13) regulation of AdoMet decarboxylase by putrescine determines the overall rate of polyamine synthesis. 

Hi) Poly amines. In many respects the role of poly amines in plant functioning is still mysterious after many years work. They are almost certainly involved in the control of growth and development through their interactions with nucleic acids and membranes (Smith, 1985). There is increasing circumstantial evidence for their involvement, especially of putrescine, in plant responses to a wide range of stresses including pH, Mg deficiency, osmotic shock, cold, SO2 pollution, and cadmium and ammonium toxicity (Smith, 1985). It remains to be determined, however, how, and indeed whether, putrescine accumulation in response to these diverse stresses is beneficial. 

Kamei et al. [45] separated spermine, spermidine, putrescine, and cadav-erine in an ion-pair reversed-phase LC system and detected the hydrogen peroxide formed in the reaction catalyzed by the enzymes putrescine oxidase and polyamine oxidase with POCL. The same analytes were determined in a later study [46], together with the acetyl derivatives. The sensitive determination of uric acid, selectively converted to hydrogen peroxide by uricase, has been investigated by several authors [37, 47],... 

Interlaboratory studies were carried out on the precision characteristics of the analytical methods used for determination of certain biogenic amines in fish and fish products, as required by German law. These included putrescine (4a), cadaverine (4b), tyramine (5) and histamine (6)78. 

Noto T, Hasegawa T, Kamimura H, Nakao J, Hashimoto H, Nakajima T. 1987. Determination of putrescine in brain tissue using gas chromatography-mass spectrometry. Anal Biochem 160 371-375. 

Rogers and Staruszkiewicz (1997). Collaborative study-GLC determination of cada-verine and putrescine in seafood fluorometric method for histamine in tuna and mahimahi, J. AO AC, 80, 591. 

Ornithine or a, h-diamimrvalerianic Acid.—In 1877 JafTe obtained from the urine of birds, which he had fed with benzoic acid, dibenzoyl ornithine or ornithuric acid, and from this substance he prepared ornithine chloride. He regarded it as a diaminovalerianic acid, the first known representative of the series of diamino acids, but only in 1898 was the position of the two amino groups definitely determined by Ellinger, who obtained putrescine from it by putrefaction the identity of putrescine with tetramethylenediamine had been previously shown by... 

Experiments utilizing D,L-a-difluoromethylornithine (DFMO), an irreversible inhibitor of ornithine decatboxylase, to determine its effect on mitogen- or al-loantigen-induced lymphocyte proliferation and CTL induction have certain similarities to the inhibition seen in N=0-induced inhibition of lymphocyte function. Stimulation of both mouse and human T cells in the presence of DFMO results in partial inhibition of proliferation which can be reversed by addition of the polyamines putrescine, spermidine, and spermine (Bowlin et al., 1987 McCarthy et al., 1990). lL-2 levels in these lymphocyte cultute supernatants as well... 

Hornero-Mendez and Garrido-Fernandez (19) proposed a method for determining BAs in fermented vegetables. The BAs are extracted using the Hui and Taylor (89) method (slightly modified to adapt it to the matrices considered) and then derivatized with DNS and analyzed by HPLC-DAD. Putrescine, Cad, and Tyr were found to be present in the olive samples considered... 

Sugiura, T., Hayashi, T., Kawai, S., and Ohno, T., High speed liquid chromatographic determination of putrescine, spermidine and spermine, J. Chromatogr., 110, 385, 1975. 

The rate of deamination is determined by the chain length and with polymethylene diamines, NH2-(CH2)n-NH2, the maximum rate occurs when n = 4 (putrescine) and n = 5 (cadaverine) and decreases to zero when n = 9 or more. At this chain length, MAO becomes active and can oxidatively deaminate the diamines. It should be noted that although MAO can deaminate both substituted and primary amines, DAO can deaminate only primary amines. 

Cells treated with 5-FU undergo cell cycle arrest or apoptosis by inhibition of DNA synthesis. When human oral squamous cell carcinomas (HSC-2, HSC-3, HSC-4) were treated with 5-FU, viable cell numbers declined dose-dependently (CC50 determined after 24 hours treatment 3.4,6.9 and 1.7 p,M, respectively). The intracellular putrescine concentration slightly declined (Table 2) [29]. The combination treatment of 5-FU with N(l),N(ll)-diethylnorspermine (DEN-SPM), which is an inducer of spermidine/spermine N(l)-acetyltransferase (SSAT) that depletes polyamine, has been reported to augment the cytotoxic activity of 5-FU, suggesting possible clinical application [30]. 

MK-2 and GG did not apparently affect the putrescine levels. This further supports the suggestion that these compounds induce non-apoptotic cell death, although the type of cell death has not yet been conclusively determined. 

In the assay described by Beeman and Rossomando (1989), a /xBondapak CI8 column (3.9 mm x 300 mm) was used to separate L-[2,3-3H]ornithine from [1,2-3H]putrescine. The mobile phase contained 0.05 M sodium phosphate (pH adjusted to 3.9 with phosphoric acid) containing 0.01 Af SDS and 36% acetonitrile. Fractions were collected and the radioactivity determined by a liquid scintillation counter The flow rate was 1.0 mL/min, and 0.5 mL fractions were collected. 

Figure 9.55 HPLC separation of the benzoyl derivatives of putrescine (PT) and spermidine (SPD) contained in a standard reaction sample and in the corresponding blank after 60 minutes of incubation at 37°C. Fractions (0.8 mL) were collected for determination of radioactivity. The arrows indicate a change in detector sensitivity from 0.2 to 0.05 absorbance unit full scale. (From Porta et al., 1981.)...

Another labelling agent suitable for determination by HPLC of histamine, tyramine, phenylethylamine, tryptamine, cadaverine, putrescine, spermidine and spermine in wines is 4-(4-dimethyl-aminophenylazo)benzenesulfonyl (dabsyl) chloride (see Figure 4.14) (Bockhardt et al., 1996 Romero et al., 2000). 

FIGURE 9.10. Evaluation of the peak purity of the putrescine-tryptamine system, (a) Experimental data set (b) Determination of the number of components by SVD (c) Study of impurities by window factor analysis (arrows indicate the time points at which spectra have been taken to be used as initial estimations and (d) Results of the resolution of the data set by MCR-ALS. 

Calystegins A3, Bi and B2 (257a-c) were isolated from Calystegia sepium. These alkaloids were also present in Convolvus arvenis and Atropa belladonna, and they were found to be catabolized by Rhizobium meliloti strain 41 [584], The structures of 257a-c were determined with HRMS, and H and NMR, and were confirmed by the synthesis and NMR analysis of model compounds [585], C-Putrescine served as a biosynthetic precursor for the calystegins [586],... 

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