Plasma Polyamine Research Articles

Polyamine levels in gynecologic malignancies.

Polyamines are closely related to many aspects of cell growth. Since increased amounts of polyamines in the urine of human cancer patients were reported in 1971, polyamines have been studied from the standpoint of tumor markers. In this study, polyamines in erythrocytes, plasma and urine were determined in 42 controls and 105 patients with gynecologic malignant tumors. The changes in polyamine levels were investigated before and after treatment. With advances in the stage of uterine cervical cancer, the frequency of abnormal levels of polyamines (concentrations greater than two standard deviations above the mean control level) became greater, and reached nearly 80% in recurrent and ovarian cancer. In the early stage of cancer, the diagnostic value was low. Comparison with carcinoembryonic antigen (CEA) was also performed. The polyamines lack specificity for malignant diseases, but they can be used to some extent as a tumor marker in the gynecologic field.

Determination of polyamines in plasma and erythrocytes of patients with chronic renal failure

Determination of polyamines in plasma and erythrocytes of patients with chronic renal failure

A rapid gas chromatographic method for the determination of plasma polyamines and its application to the prediction of tumour response to chemotherapy

Isobutyloxycarbonyl derivatives of the polyamines, spermidine, spermine and putrescine in 1 ml plasma were analysed on a mixed phase, 1.5% SE-30, 0.15% FFAP, column by temperature-programmed gas chromatography using a nitrogen-sensitive glass bead detector. Levels in 60 normal subjects (mean ±2 S.D.) were 0.13 ± 0.08 μmol/l for spermidine, 0.02 ± 0.04 for spermine and ⩽ 0.18 for putrescine. Elevated spermidine levels were found pre-treatment in only 20% of patients with advanced metastatic cancer, reflecting the lack of sensitivity of polyamines as markers for malignant disease. However, serial studies in 26 patients undergoing remission induction chemotherapy showed that a significant rise in plasma spermidine within 48 h correlated with subsequent clinical response ( p < 0.001). Tumour response to chemotherapy can be predicted by the assay of plasma polyamines.

Plasma polyamine levels in patients with liver insufficiency.

Relevant statistical approaches demonstrated a normal distribution of the values for the healthy control group and an abnormal distribution in the diseased group. Within both groups the polyamines were found to be mutually linearly correlated with the exception of the putrescine — spermine pair in the normals and the diaminopropane/spermine pair in the patients. A comparison between the diseased and healthy groups demonstrated significant elevations of putrescine, cadaverine, and spermidine in the former. In 78% of the patients, at least one of the polyamines was found to be abnormally elevated, whereas no elevations whatsoever were observed in 22% of the cases. Putrescine, at concentrations up to 3 / , and cadaverine up to 6 / showed the highest elevations in comparison to normals (85.3 ± 35.3 nmol/1 for putrescine and lower than 20 nmol/1 for cadaverine). Spermidine, spermine and diaminopropane concentrations reached concentrations of approximately 1 >1/1, 500 nmol/1 and 150 nmol/1, compared with the mean normal values of 94.6 ± 34.5 nmol/1, 39.8 ± 29.2 nmol/1, and 16.9 ± 9.31 nmol/1, respectively. In patients, cadaverine is inversely correlated with the liver normotest, and spermine is directly correlated with cholinesterase activity. Liver involvement is discussed with regard to the Value of polyamine levels as tumour markers. No definite conclusions can be drawn as to the cause of the polyamine elevations found in liver-diseased patients.

Raised polyamines in erythrocytes from melanoma-bearing mice and patients with solid tumours.

The levels of polyamines (putrescine, spermidine and spermine) in erythrocytes and plasma were studied using Cloudman S-91 melanoma grown in the lungs of DBA/2 mice. Polyamine levels and the numbers of tumour-cell colonies in the lungs were determined at weekly intervals. Putrescine levels in both erythrocytes and plasma significantly increased 1 week after tumour inoculation. Three weeks after inoculation, however, putrescine levels in the erythrocytes showed a greater increase than those in plasma. Spermidine and spermine levels were initially high at 2 weeks in plasma and at 4 weeks in erythrocytes. However, by 6 weeks the spermidine levels showed a greater increase in erythrocytes than in plasma. These data suggest that erythrocytes may absorb and store polyamines released into the circulation.This finding was subsequently applied to human studies. Fifty-two untreated patients with solid tumours were examined in the preoperative period. All erythrocyte polyamine levels from patients were significantly higher than those from control subjects. Plasma spermidine levels in patients were significantly higher than those in controls, whereas plasma putrescine and spermine levels showed no significant increase. The frequency of raised levels of putrescine, spermidine and spermine in erythrocytes was significantly greater than in plasma. These results suggest that polyamine levels in erythrocytes may provide useful information for the detection of cancer.

Inhibition of Lymphocyte Proliferation by Polyamines Requires Ruminant-Plasma Polyamine Oxidase

Spermine and spermidine in vitro are potent inhibitors of proliferation of phytohaemagglutinin-stimulated rat thymic lymphocytes, lymphoma cells and human lymphoblastic leukaemia cells, but only in media supplemented by foetal calf serum. This inhibition is shown to be due to a bovine plasma polyamine oxidase, with a high specificity for these polyamines. Spontaneously dividing lymphocytes are not subject to this inhibition. This, plus direct evidence from synchronous cultures of EB2 cells demonstrates that the inhibition is expressed in the late G1 or G1/S interface of the cell cycle. Putrescine was not an inhibitor in the presence of foetal calf serum but became so in the presence of human pregnancy serum, possibly due to the action of diamine oxidase.

CORTICOTROPHIN INCREASES CEREBRAL POLYAMINE CONTENT

Abstract— To determine whether changes in cerebral polyamines might mediate previously reported ACTH‐induced changes in brain biochemistry and behavior, the cerebral content of polyamines was examined following ACTH treatment. Male CD‐1 mice were injected daily for 3 days with long‐acting (zinc phosphate) preparations of ACTH1–24 (1 μg/g) or ACTH4–10 (0.33 μg/g) and killed 24 h after the last injection. Putrescine, spermidine and spermine contents were determined by high pressure liquid chromatography. Putrescine content was significantly elevated in all brain regions by ACTH1–24 (approx 50%), and in the telencephalon by ACTH4–10 At the dose tested ACTH4–10 was less effective than ACTH1–24. Telencephalic spermidine was also elevated (10%)by ACTH1–24, but spermine content was not altered in any brain region. One injection of the long‐acting ACTH1–24 preparation elevated telencephalic putrescine (49%) 24 h post‐injection. ACTH1–24 (1 μg/g) in saline produced a peak elevation of all three telencephalic polyamines 6 h post‐injection, while in the liver only putrescine was significantly elevated and reached a peak at 10h. Neither plasma polyamine nor ornithine concentrations were significantly altered by any of the treatments. Corticosterone, in both single and multiple injection regimens, failed to alter telencephalic polyamine content. Adrenalectomy, however, prevented the ACTH1–24‐induced increase in telencephalic polyamines. It is concluded that ACTH acts directly in the brain to increase cerebral polyamine concentrations. The possibility that adrenal hormones exert permissive effects on this action is discussed.

Plasma polyamines determined by negative-ion chemical ionization/mass spectrometry.

We have developed an accurate and highly sensitive gas-chromatographic/mass-spectrometric procedure for determining di- and polyamines (putrescine, spermidine, and spermine) in plasma and erythrocytes. Deuterium-labeled analogs of putrescine, spermidine, and spermine were synthesized for use as internal standards. Trifluoroacetyl derivatives of the polyamines were formed during the procedure and then detected with negative-ion chemical ionization/mass spectrometry in combination with multiple ion monitoring. Limits of sensitivity ranged from 0.25 to 1.0 pmol of analyte injected into the instrument.

Polyamines in amniotic fluid, plasma, and urine during normal pregnancy

Polyamines have been implicated as markers of cell kinetic parameters. Putrescine and spermine were elevated in the urine of women with normal pregnancies. Particularly striking was the over 75-fold increase in spermine excretion. Putrescine was more than twofold that found for normal women. In several patients followed serially during pregnancy to assess the temporal pattern of the urinary excretion of polyamines, it was found that the highest levels of all three polyamines occurred at 12 weeks of gestation. In amniotic fluid, putrescine was higher prior to 30 weeks of gestation whereas spermidine was significantly higher at or beyond 30 weeks of gestation. Spermine was relatively high in both groups. Studies are ongoing to assess whether alterations from these normally elevated patterns will occur in the fluids of pregnant women at high risk for abnormal fetal development.

892 HUMAN PLASMA POLYAMINE-CONJUGATED PEPTIDE, PUTRESCINI

To define a cystic fibrosis factor and its potential relationship to polyamine metabolism an investigation of the low molecular weight peptide profile of human blood was undertaken. Previous investigations indicated polyamines were detectable in serum and plasma following extensive alkali or acid hydrolysis. We report the existance of a polyamine peptide conjugate in human plasma. This peptide was isolated by fractionation of citrated whole human plasma following chromatography on Biogel P10. The elution profile, monitored at 220nm, resulted in the identification of 6 peaks. These polypeptides had a molecular weight range of 2 to 10,000 daltons as evidence by retention on Diaflomembranes and gel chromatography. The major peak (putrescinin) was purified by Biogel P6 chromatography, paper chromatography and DEAE celulose chromatography. This plasma peak was present in CF patients and controls, and disappeared from blood following coagulation. The molecular weight of putrescinin was 4600 daltons. Amino acid analysis revealed 43 amino acid residues with absence of methionine, cysteine and abundant quantitites of glycine, glutamate asparate, serine and alanine. One mole of ornithine was detected per mole of putrescinin. Putrescine and spermidine was covalently bound to putrescinin; 3-5 moles of putrescine and 0.3-0.5 moles of spermidine/mole peptide. The conjugated polyamine content was 10-30 times that of free plasma polyamine. The significance of this peptide in coagulation and to CF will be discussed.

The content of unbound polyamines in blood plasma and leukocytes of patients with polycythemia vera

In the blood plasma and isolated leukocytes of 11 patients with polycythemia vera and 3 healthy subjects, the polyamines putrescine, spermidine and spermine were determined. The average values in the leukocytes of the healthy volunteers were found to be 1.8 ± 1.4 nmol putrescine/10 8 cells, 3.0 ± 0.9 nmol spermidine/10 8 cells and 12.9 ± 3.8 nmol spermine/10 8 cells. In the plasma of healthy persons the amounts of the polyamines were below the sensitivity level of the method employed. In 4 patients with polycythemia vera no polyamines were detected. In contrast, in 7 cases 0.1 to 3 nmol polyamines/ ml were found. The level of polyamines in the leukocytes of 6 of these patients was decreased and in one patient corresponded to the values found in the normal range (17.7 ± 6.0 nmol polyamines/10 8 cells). Continued blood-letting therapy on 3 patients led to values approaching the concentrations found in normal subjects in both blood plasma and leukocytes. A decreased amount of these diamines in the leukocytes of the patients was seen to correlate with an elevated concentration in the plasma.

Relative Usefulness of Measuring Polyamines in Serum, Plasma, and Urine as Biochemical Markers of Cancer

Serial samples of plasma and serum were collected in the morning and afternoon from cancer patients, along with 24-h urine specimens. Values for serum and plasma samples taken at the same time from the same patient differed little, suggesting that either procedure is acceptable for polyamine analysis. Increases in concentrations of putrescine and spermidine in serum and plasma correlate well with such increases in 24-h urine samples. Spermidine concentrations in sera were consistently about 10-fold lower than corresponding urine values. Putrescine concentrations were 10- to 100-fold different. The variation in putrescine values may be due to its more active metabolism by diamine oxidase, an enzyme known to be present in serum. Concentrations of polyamines in serum and urine increased in response to effective chemotherapy.