To determine the structure(s) of the spermidine conjugate excreted into urine, extensive pharmacokinetic studies on the turnover of [¹⁴C]spermidine were conducted in both rats and humans. These studies demonstrated that exogenously administered [¹⁴C]spermidine equilibrated with endogenous polyamine pools. Radiolabeled urine collected from the humans was subjected to a cleanup protocol and subsequent analysis by GC-MS, which demonstrated the presence of N-acetylspermidine. Further analysis of the radiolabeled urine by thin layer chromatography (TLC) demonstrated the presence of both N¹- and N⁸-acetylspermidine in an approximate 1 to 1 ratio. Using similar methodologies the monoacetyl derivatives of putrescine and cadaverine were found to be the primary conjugated products of these polyamines excreted into human urine. Radiolabeled rat tissue extracts, analyzed by TLC, demonstrated that all tissues studied contained [¹⁴C] N¹- and N⁸-acetylspermidine. N¹-acetylspermidine was the primary isomer detected in all tissues; although, N⁸-acetylspermidine was detected in all tissues studied. The N-acetylspermidine content of an isolated cell system was determined in Chinese hamster ovary cells. These cells contained both N¹- and N⁸-acetylspermidine in an approximate 2 to 1 ratio. To directly measure the N-acetylpolyamines excreted into urine, two high performance liquid chromatography (HPLC) methods were developed. Both HPLC methods utilize a cation exchange resin, one using high pH, low salt buffers and the other low pH and high salt buffers. The primary N-acetylpolyamine excreted into human urine is N-acetylputrescine with lesser amounts of N¹- and N⁸-acetylspermidine which exist in a 1 to 1 ratio. In contrast, cancer patients excreted elevated amounts of both N-acetylputrescine and N¹-acetylspermidine. Cystic fibrosis patients were also found to excrete elevated amounts of N¹-acetylspermidine resulting in a consistently elevated N¹- to N⁸-acetylspermidine ratio. Mice injected with P-388 leukemia tumor excreted elevated amounts of N-acetylputrescine, N¹-acetylspermidine and N⁸-acetylspermidine. In contrast, the excretion of the unconjugated polyamines putrescine and spermidine in these animals was decreased, suggesting that the altered polyamine excretion was not primarily due to the presence of the tumor. Administration of Adriamycin to the tumor bearing animals resulted in the elevation of N¹-acetylspermidine excretion which was proportional to the relative tumor burden. Similar results were obtained from 2 human leukemia patients studied following chemotherapy.

No description available.

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The main aims were to: optimise the method for extracting polyamine from food and blood products; study the effect of diet (after oral loading with orange juice – a food source rich in polyamines) on blood polyamine concentrations and urinary polyamine excretion; assess the concentrations of S100β protein, NSE, GFAP and polyamines in healthy control populations; study the time profile or kinetics of polyamines, NSE, S100β protein and GFAP after acute ischaemic stroke and their relationship with stroke severity and outcome using NIHSS, mRS and BI; and study the relationship between risk factor burden, aetiological mechanisms and clinical stroke sub-types and biomarker trends. Several steps were taken to improve the quality of the acid soluble extract from red blood cells.  This led to a highly reproducible and high quality extract for HPLC analysis.  Evidence gathered from the results confirmed the absorption of the content of orange juice but this did not reflect in changes in blood polyamine concentrations.  Spermidine and spermine show significant correlation with stroke severity and functional outcome comparable to S100β protein and better than NSE and GFAP. The number of participants involved in the study is the largest ever used for any clinical assessment of polyamines in stroke but still falls short for a thorough assessment of these markers.  Challenges posed by shortage of reagents for the HPLC run remained largely unresolved until recently. Limited follow-up in the stroke study restricted the evaluation of the polyamines to short term outcome measures.

Thesis (M.S.)--Virginia Polytechnic Institute and State University, 1988. / Vita. Abstract. Includes bibliographical references (leaves 51-60). Also available via the Internet.

Many grades of paper have shifted from acid to alkaline production conditions. This change excludes the usage of traditional alum-rosin sizing chemistry. In some cases however, rosin sizing could prove beneficial if it could be used under neutral to alkaline paper making conditions. One effort toward this goal has been the use of epi-halohydrin modified polyamine/polyaminoamide in place of alum. The optimum sizing conditions, as well as the mechanism that modified polyamines/polyaminoamides use to size rosin, is investigated. Polyethyleneimine modified with epi-chlorohydrin was used in these studies. Rosin dosage was found to be the main variable that controlled the degree of sizing achieved. As the only hydrophobic material added, this was expected. In addition to rosin dosage, the polymer and pH had significant impacts on this sizing chemistry. Laboratory data shows that the synthesis procedure for and structure of polyethyleneimine-epi-cholohydrin (PEI-epi) impacts the degree of sizing achieved. Use of different modification techniques and degrees of modification illustrated several key factory regarding the polymer as a rosin sizing mordant. Prevention or removal of by-products from epi-chlorohydrin imporves the sizing efficiency of PEI-epi with rosin. A critical molecular weight of the PEI-epi was found to be important to sizing. Higher charge density on PEI-epi provides better the sizing efficiency. In addition to the polymer, other important factors where examined. The pH of the stock impacted the degree of sizing and permanency of the sizing achieved. Varying stock pH showed changes in sizing that are consistent with effects seen for rosin retention and of PAE as a wet strength agent. Low and high pH levels also caused reversion of the sizing effect. A pilot trial of the PEI-epi-rosin sizing chemistry was conducted at the Herty Foundation. The pilot trial shows that PEI-epi-rosin can reach the required level of sizing for linerboard. Trial results showed that calcium carbonate filled sheets could be sized by this chemistry. No significant impact on strength is seen from the sizing chemistry at the required dosage levels. As would be expected, addition of filler diminishes sheet strength. No sizing reversion was seen from any samples of this trial. Size reversion at low stock pH indicated that acid catalytic cleavage of an ester bond could have been occurring. Addition of acid or base to permanently sized sheets showed significant levels of reversion, similar to that seen for low stock pH sheets. This provides further supporting eviden ce for the formation and importance of an ester bond between rosin and PEI-epi. FTIR spectra were collected for model compounds to further examine the presence and importance of ester bonds. An ester bond was found to form at room temperature with aging or immediately with drying. Further, the ester bonds were found to decrease when the sample was subjected to acidic or basic environments. Solid state NMR was run to demonstrate the presence and importance of ester bonds to sizing in handsheets. Use of a [superscript 13]C labeled fatty acid showed that the ester bond peak dominated when sizing was present. After treatment with sodium hydroxide, the sizing disappeared and much of the ester bond signal shifted to a carboxylic acid or salt. Further solid state NMR of a non-reactive cationic polymer or a fully pre-reacted PEI-epi polymer shows retention of the fatty acid, but no ester bond and no sizing. The sizing mechanism of PEI-epi-rosin sizing appears to be connected to an ester bond formed during drying or with aging. While the amount of rosin retained controls the degree of sizing, retention alone is not enough to give sizing. The most effective pH for this sizing chemistry is near neutral, pH 6-8. FTIR and solid state NMR shows that an epichlorohydrin modified polyamine can form ester bonds with fatty acids in handsheets under conditions comparable to papermaking. The presence of residual acid or base in handsheets destroys the sizing effect achieved. Destruction of the sizing by addition of sodium hydroxide parallels the cleavage of many of the ester bonds present. PEI-epi appears to function as a mordant for rosin by creating an ester bond to anchor and help orient the hydrophobic portion of rosin.

Polyamine titer in the leaves of the self-seedlings of Doritis pulcherrima Lindley¡]cv. S84-3352¡^under short-day ( SD, 9 hours of light period ) and long-day ( LD, 16 hours of light period ) exposures were determined. A significant reduction in acid insoluble spermidine levels occurred in D. pulcherrima leaves after 30 days of SD compared to the LD exposures. After 30 days of SD there was a transient decrease in acid insoluble spermidine and an increase in acid insoluble spermine in the leaves. When the flowering shoot reached 7 -10 cm long ( with flower primordia in SD ), free cadaverine, acid insoluble putrescine, and spermine levels were higher than those in 2-3 cm flowering shoot ( with no flower primordium derived from SD ) These results suggest that polyamines are implicated differently during photoperiodic flower induction and flower devlopment in D. pulcherrima.

Thesis (M. Phil.)--University of Hong Kong, 1986.

published_or_final_version / Biochemistry / Master / Master of Philosophy

No description available.