On Ornithine Decarboxylase Activity Research Articles

Effect of remedies for enhancing resistance and relieving blood stasis on metastasis in postoperative gastric cancer and ornithine decarboxylase levels.

To study the action of remedies for enhancing resistance and relieving blood stasis on metastasis in postoperative gastric cancer and its influence on ornithine decarboxylase (ODC). Sixty-three postoperative patients with gastric cancer were randomly divided into two groups. Thirty-one patients were treated with western medicine consisting of the FAP (5-fluorouracil, adriamycin, cisplatin) and CODP regimens (cyclophosphamide, vincristine, daunorubicin, prednisone), whereas 32 patients were treated with the FAP regimen and traditional Chinese medicine. Correlations were made between the ODC levels detected before and after treatment and other factors such as tumor diameter, infiltration depth, histological type, and lymph node metastasis. The ODC levels in the gastric cancer tissue and adjacent normal gastric mucosal tissue were significantly higher in the patients than in the controls. There was an obvious correlation between increased ODC and tumor size, infiltration depth, degree of differentiation, and lymph node metastasis. Six months later, there were no significant changes in the ODC levels of the group using only Western medicine, while the ODC levels decreased markedly in the group using combined Western and traditional Chinese medicine (P < 0.01). The effects of traditional Chinese medicine remedies on metastases in postoperative gastric cancer are related to the reduction of ODC activity.

Effects of exposure to DAMPS and GSM signals on ornithine decarboxylase (ODC) activity: I. L-929 mouse fibroblasts

Purpose: A temporary increase in ornithine decarboxylase (ODC) activity was reported in lysed L-929 fibroblasts after exposure to the microwaves emitted by Digital Advanced Mobile Phone System (DAMPS-835 MHz, 2.5 W/kg, 8 hours). Confirmation of these data was undertaken, given the suggested potential physiopathological consequences, i.e., tumour promotion.Materials and methods: Murine L-929 fibroblasts were exposed at various Specific Absorption rates (SAR) to (DAMPS) or Global System for Mobile communications (GSM) signals using different set-ups. Cell ODC activities were assayed using 14CO2 generation from 14C-labeled L-ornithine.Results: ODC activity in live L-929 cells showed no significant alteration after exposure at an SAR of 2.5 W/kg, for one hour at the end of exposure to 50 Hz-modulated DAMPS-835 using Transverse Electro-Magnetic (TEM) cells. No significant alteration in ODC activity was observed at 6 W/kg, with active fans to regulate temperature (37°C). Tests using cell lysed after exposure in another temperature-controlled set-up (waveguides) did not confirm the published studies reporting increased ODC activity in Radio-Frequency radiation (RFR)-exposed L-929 cells. In the second part of the study, no alteration of ODC activity was detected when L-929 cells were exposed to different RFR signals: 217 Hz modulated GSM-900 (wire-patch antenna) or GSM-1800 (waveguides), and lysed before ODC measurement.Conclusion: We conclude that under our exposure conditions, DAMPS-835 and GSM signals have no influence on ODC activity in L-929 cells.

Effects of exposure to DAMPS and GSM signals on Ornithine Decarboxylase (ODC) activity: II. SH-SY5Y human neuroblastoma cells

Purpose: An increase in Ornithine Decarboxylase (ODC) activity was reported in L929 murine fibroblast cells after exposure to a digital cellular telephone signal. This result was not confirmed by several other studies, including the one reported in a companion paper. As a partner in the Perform-B programme, we extended this study to human neuroblastoma cells (SH-SY5Y), using well-defined waveguide systems to imitate exposure to radiofrequency radiation (RFR): Digital Advanced Mobile Phone System (DAMPS) or Global System for Mobile communications (GSM) signals emitted by mobile phones.Materials and methods: Human neuroblastoma cells (SH-SY5Y) were exposed at various Specific Absorption Rates (SAR) to DAMPS or GSM signals using different set-ups. Cell ODC activities were assayed using 14CO2 generation from 14C-labeled L-ornithine.Results: SH-SY5Y cells were incubated for 20 hours, and were blindly exposed to 50 Hz-modulated DAMPS-835 or 217 Hz-modulated GSM-1800 for 8 or 24 h using Information Technologies in Society (IT'IS) waveguides equipped with fans. After cell lysis, ODC activity was determined using 14C-labeled L-ornithine. ODC activity was estimated by the 14CO2 generated from 14C-labeled L-ornithine, as generated d.p.m. 14CO2/h/mg protein. The results showed that, irrespective of the signal used (835 MHz/DAMPS, or 1800 MHz/GSM) and exposure conditions (duration and SAR), human SH-SY5Y neuroblastoma cells did not exhibit any alteration in ODC enzyme activity.Conclusion: This work did not show a significant effect of mobile phone RFR exposure on ODC activity in neuroblastoma cells (SH-SY5Y).

Bursera fagaroides, effect of an ethanolic extract on ornithine decarboxylase (ODC) activity in vitro and on the growth of Entamoeba histolytica

The effect of an ethanolic extract from the stem bark of Bursera fagaroides on ornithine decarboxylase (ODC) activity in vitro and on the growth of Entamoeba histolytica was evaluated. For this purpose, increasing concentrations of the extract, up to 8.0 mg/mL, were added to amoeba cultures or ODC reaction mixtures, which were incubated at 37 °C. Metronidazole and G418 were added as controls. After 1.5 and 72 h, the ODC activity in vitro and growth, respectively, were determined. Results revealed a strong inhibition of growth with IC 50 values on the order of 0.05 mg/mL. ODC activity, on the other hand, was inhibited by 12% and 50% at concentrations of 4.0 and 8.0 mg/mL, respectively.

Astroglia growth retardation and increased microglia proliferation by lithium and ornithine decarboxylase inhibitor in rat cerebellar cultures: Cytotoxicity by combined lithium and polyamine inhibition

Lithium, the most prevalent treatment for manic-depressive illness, might have a neuroprotective effect after brain injury. In culture, lithium can exert neurotoxic effects associated with reduction in polyamine synthesis but neuroprotective effects as cultured neurons mature. Cumulative evidence suggests that lithium may exert some of its effects on neurons indirectly, by initially acting on glial cells. We used rat cerebellar cultures to ascertain the effects of lithium on ornithine decarboxylase (ODC) activity, the enzyme catalyzing the first step in polyamine synthesis, and to compare effects of lithium with those of the ODC inhibitor alpha-difluoromethylornithine (DFMO) on neuron survival and glial growth. Switching cultures from high (25 mM) to low (5 mM) KCl concentrations served as the traumatic neuronal insult. The results indicate the following. 1) Whereas high depolarizing KCl concentration enhances neuron survival, it inhibits astroglial growth. 2) Lithium (LiCl; 1-5 mM) enhances neuronal survival but inhibits astroglial growth. 3) Lithium treatment leads to reduced ODC activity. 4) DFMO enhances neuron survival but inhibits astroglial growth. 5) Lithium and DFMO lead to transformation of astroglia from epithelioid (flat) to process-bearing morphology and to increased numbers of microglia. 6) Combined lithium plus DFMO treatment is cytolethal to both neurons and glia in culture. In conclusion, lithium treatment results in growth retardation and altered cell morphology of cultured astroglia and increased microglia proliferation, and these effects may be associated with inhibition of polyamine synthesis. This implies that direct effects on astrocytes and microglia may contribute to the effects of lithium on neurons.

Effects of cyclosporin and ultraviolet radiation on growth and ornithine decarboxylase activity in cultured human epidermal keratinocytes.

Both cyclosporin (CyA) and ultraviolet radiation are effective in the treatment of psoriasis, but their precise mechanisms of action are uncertain. We investigated their effects on ornithine decarboxylase (ODC) activity, ODC gene expression, and cellular proliferation stimulated by epidermal growth factor (EGF), in cultured normal human epidermal keratinocytes. CyA (5 micrograms/ml) inhibited ODC activity, ODC mRNA level, and cell growth induced by 50 ng/ml EGF. Ultraviolet B (10 mJ/cm2) irradiation suppressed the induction of ODC, ODC mRNA, and cell proliferation stimulated by EGF, but ultraviolet A (0-15 J/cm2) irradiation inhibited neither EGF-stimulated ODC activity nor cell proliferation. These findings indicate that reduction of ODC activity in CyA- or ultraviolet B-treated human keratinocytes may contribute to the antiproliferative mechanism of these agents. These results also suggest that the regulation of ODC activity by ultraviolet B and A irradiation may be mediated by different signal transduction pathways.

Modest increase in temperature affects ODC activity in L929 cells: low-level radiofrequency radiation does not

The effects of low-level radiofrequency (RF) radiation and elevated temperature on ornithine decarboxylase (ODC) activity were investigated in murine L929 fibroblasts. The cells were exposed at 900 MHz either to a pulse-modulated (pulse frequency 217 Hz; GSM-type modulation) or a continuous wave signal at specific absorption rate (SAR) levels of 0.2 W kg(-1) (0.1-0.3 W kg(-1)) and 0.4 W kg(-1) (0.3-0.5 W kg(-1)) for 2, 8, or 24 h. RF radiation did not affect cellular ODC activity. However, a slight increase in temperature (0.8-0.9 degrees C) in the exposure system lead to decreased ODC activity in cell cultures. This was verified by tests in which cells were exposed to different temperatures in incubators. The results show that ODC activity is sensitive to small temperature differences in cell cultures. Hence, a precise temperature control in cellular ODC activity studies is needed.

Hexachlorobenzene-induced early changes in ornithine decarboxylase and protein tyrosine kinase activities, polyamines and c-Myc, c-Fos and c-Jun proto-oncogenes in rat liver.

Hexachlorobenzene (HCB) is a lipophilic chemical compound that is widely distributed in the environment. HCB is known to cause liver tumors in experimental animals. In the present study the in vivo effect of HCB treatment on ornithine decarboxylase (ODC) and protein tyrosine kinase (PTK) activities, free polyamine content, and c-Myc, c-Fos, and c-Jun protein levels in rat liver were investigated. HCB (1000 mg/kg body weight) increased hepatic immunodetectable c-Myc, c-Fos, and c-Jun levels after 6 h, and ODC activity and spermine and putrescine content after 18 and 24 h, while maximum stimulation of PTK activity occurred at 12 h. PTK and ODC activities varied in a dose-dependent manner. The time-course of c-Myc, c-Fos, and c-Jun protein levels was different for each proto-oncogene. They were all elevated at the second day of treatment, while only c-Fos and c-Jun remained elevated after 10 days of HCB exposure. These data jointly suggest that the increase in ODC activity may be the consequence of proto-oncogene induction. The alterations in PTK activity suggest that the growth factor signal transduction pathway may be involved in the regulation of the proto-oncogene levels or/and ODC activity. The decrease in PTK activity after the first day, even in the presence of alpha-D-Difluoromethylornithine (DFMO), an inhibitor of ODC activity, suggests that it is not regulated by polyamines. These results may be relevant to the early molecular events involved in HCB tumor promoter activity in rat liver.

Whole-protein-based enteral formula stimulates intestinal ornithine decarboxylase activity more than single amino acids but does not affect mucosal adenosine triphosphate content in early postsurgical refeeding.

To restore intestinal integrity after starvation and trauma, luminal nutrients are essential. Specific nutrients such as glutamine support mucosal proliferation and energy metabolism. The aim of this study was to compare effects of enteral formula vs specific amino acids on ornithine decarboxylase (ODC) activity and adenine nucleotide metabolism in jejunal mucosa. Male Wistar rats (240 to 280 g) were starved for 48 hours and subjected to intestinal transection, gastrotomy, and jejunal instillation of 5mL nutrient solution. In the first experiment, standard enteral formula (EF) was compared with isonitrogenous formula supplemented with the glutamine precursor, alpha-ketoglutarate (alpha-KG). In a second experiment, 2% glutamine was compared with isonitrogenous ornithine alpha-KG, arginine alpha-KG, glycine and diluted standard enteral formula (EF), or saline. The ODC activity, adenosine triphosphates (ATP), and RNA and protein in the jejunal mucosa were analyzed 2 hours after surgery. The ODC peak in jejunal mucosa in animals treated with EF was higher than when supplemented with alpha-KG (p < .05). Compared with specific amino compounds, EF resulted in a significantly higher ODC peak and no differences were seen between the different specific amino acids. Differences seen in ATP or energy charge between the groups were not significant. Immediate postoperative enteral feeding by standard EF results in rapid increase of ODC activity. This response was attenuated when the enteral nutrition was supplemented with alpha-KG and was absent when isonitrogenous single amino acids were administered. We found no significant effects on ATP content in the small bowel mucosa by supplementing the diet with alpha-KG.

Epidermal ornithine decarboxylase and polyamines in mice exposed to 50 Hz magnetic fields and UV radiation

We studied the influence of magnetic fields (MFs) and simulated solar radiation (SSR) on ornithine decarboxylase (ODC) and polyamines in mouse epidermis. Chronic exposure to combined MF and SSR did not cause persistent effects on ODC activity or polyamines compared to the animals exposed only to UV, although the same MF treatment was previously found to accelerate skin tumor development. In an acute 24-h experiment, an elevation of putrescine and down-regulation of ODC activity was observed in the animals exposed to a 100-microT MF. No effect was seen 24 h after a single 2-MED (minimal erythemal dose) exposure to SSR. The results indicate that acute exposure to 50 Hz MF does exert distinctive biological effects on epidermal polyamine synthesis.

Epidermal ornithine decarboxylase and polyamines in mice exposed to 50 Hz magnetic fields and UV radiation

We studied the influence of magnetic fields (MFs) and simulated solar radiation (SSR) on ornithine decarboxylase (ODC) and polyamines in mouse epidermis. Chronic exposure to combined MF and SSR did not cause persistent effects on ODC activity or polyamines compared to the animals exposed only to UV, although the same MF treatment was previously found to accelerate skin tumor development. In an acute 24-h experiment, an elevation of putrescine and down-regulation of ODC activity was observed in the animals exposed to a 100-μT MF. No effect was seen 24 h after a single 2-MED (minimal erythemal dose) exposure to SSR. The results indicate that acute exposure to 50 Hz MF does exert distinctive biological effects on epidermal polyamine synthesis. Bioelectromagnetics 19:388–391, 1998. © 1998 Wiley-Liss, Inc.

Acute inhibitory effect of excess iodide on ornithine decarboxylase in the thyroid of propylthiouracil-treated rats.

Polyamines such as putrescine, spermidine and spermine have been thought to play an important role in thyroid growth induced by goitrogens. Reduced biosynthesis of these polyamines might play a role in the antigoitrogenic effects of excess iodide. This study was designed to examine the effect of potassium iodide (KI) on ornithine decarboxylase (ODC), a rate-limiting enzyme in the biosynthesis of polyamines. Thyroidal ODC activity, protein content and mRNA were increased in rats made hypothyroid by 10 days of propylthiouracil treatment. The increase in ODC activity was suppressed after subcutaneous injection of KI (13mg/kg body weight); the apparent half-life of ODC activity after the treatment was estimated to be 19 min and the maximum suppression (90%) was seen 60 min after the treatment. On the other hand, administration of iodine-containing compounds including L-thyroxine, L-di-iodotyrosine,amiodarone, iopanoic acid and erythrosine showed no significant effect on ODC activity. The inhibitory effect of excess iodide was not reversed by pretreatment with dibutyryl cAMP and theophylline. The amount of immunoreactive ODC protein was reduced by iodide treatment (40%). However, the decrease was not as great as the decrease in ODC activity (90%). No significant change in thyroidal ODC mRNA content was seen 1 and 3 h following KI treatment. These results suggest that excess iodide reduces ODC activity in the rat thyroid gland by a post-transcriptional mechanism.

The requirement of Zn 2+ for the increase in ornithine decarboxylase induced by insulin and epidermal growth factor in primary cultured rat hepatocytes

The effect of restricted Zn 2+ on ornithine decarboxylase (ODC) was studied in primary cultured adult rat hepatocytes. ODC activity was increased 4 hr after adding insulin and epidermal growth factor. The maximal activity was maintained for 12 hr, it then decreased and returned to the control level by 24 hr. When the chelator, diethylenetriamine penta-acetic acid (DTPA) was added to insulin and epidermal growth factor, the enzyme activity was decreased by 50% and only the Zn 2+ of the bivalent metal ions tested was effective in reversing this effect. The level of the ODC messenger RNA did not change with addition of the chelator. The half-life of the ODC activity was decreased 2-fold by the chelator. Exposing the hepatocytes to DTPA reduced the incorporation of [ 3H]thymidine into acid insoluble fraction. The inhibition of DNA synthesis was fully reversed by the addition of 600 μM Zn 2+. These results suggest that a lack of Zn 2+ in hepatocytes impairs ODC activity mainly by destabilizing the enzyme resulting in the inhibition of DNA synthesis.

Inhibitory effects of d-limonene on the development of colonic aberrant crypt foci induced by azoxymethane in F344 rats.

The modifying effect of the monoterpenoid d-limonene in drinking water on the development of azoxymethane (AOM)-induced colonic aberrant crypt foci (ACF) was investigated in male F344 rats. The effects of d-limonene intake on ornithine decarboxylase (ODC) activity and on the silver stained nucleolar organizer region protein (AgNOR) count in the colonic mucosa were also estimated. Animals were given 3 weeks s.c. injections of AOM (15 mg/kg body wt) to induce ACF. These rats were treated with or without 0.5% d-limonene in the drinking water, starting 1 week before the first dosing with AOM. All rats were killed 2 weeks after the last AOM injection, to measure the number of ACF, ODC activity and AgNOR count/nucleus in the colon. In rats given AOM and d-limonene the frequencies of ACF and aberrant crypts/colon, and aberrant crypts/focus were significantly decreased compared with those of rats given AOM alone (P < 0.01, P < 0.001 and P < 0.001 respectively). Number of AgNOR counts/nucleus of rats treated with AOM and d-limonene was significantly smaller than that of rats treated with AOM alone (P < 0.001). These results suggest that the monoterpenoid d-limonene might be a chemopreventive agent for colonic carcinogenesis in rats.

Pristane-induced effects on cytochrome P-4501A, ornithine decarboxylase and putrescine in rats.

The effects of pristane (2,6,10,14-tetramethylpentadecane) on cytochrome P-4501A (cP4501A) activity in microsomes, as well as on ornithine decarboxylase (ODC) activity and concomitant putrescine levels were examined in Copenhagen rats. In general, pristane treatment led to increased cP4501A levels when compared to basal levels, while co-treatment with 3-methylcholanthrene (3-MC) and pristane elicited augmented cP4501A responses when compared to responses induced by 3-MC alone. Increases in both ODC activity and putrescine levels were also observed in pristane treated rats. Collectively, these results indicate that pristane influences cP4501A activity and elicits promoter-like responses as reflected in elevated ODC activity and increased amount of putrescine.

Effects of thyroid hormones on chick embryo muscle cell culture

The importance of thyroid hormones (TH) in the normal development of muscles has been repeatedly postulated. The effects of physiological TH doses on ornithine decarboxylase (ODC) and protein synthesis in muscle cells have been studied using cell cultures prepared from 11-day-old chick embryos. Triiodothyronine nuclear receptors in primary muscle cell culture were characterized on the basis of the specific binding analysis as a single receptor class with the equilibrium dissociation constant Kd = 1.2 +/- 0.4 x 10(-10) mol/l and binding capacity Bmax = 0.21 +/- 0.09 fmol/micrograms DNA. While the physiological amounts of both triiodothyronine (T3) and thyroxine (T4) stimulated ODC activity after 2 hr of treatment, only T3 had the same stimulatory effect after 4 hr of treatment. Twenty-four hour exposure of muscle cell culture to TH did not change ODC activity. The incorporation of [3H]leucine into proteins was elevated only after 120 hr incubation in the presence of T4. Application of T4 caused also an increase in the protein content after 24 hr.

Experimental carcinogenesis in the rat intestine induced by two different carcinogens--evaluation of carcinogenic action and its effect on ornithine decarboxylase activity.

Carcinogenesis in the rat intestine induced with 1,2-dimethylhydrazine (DMH), N-methyl-N-nitrosourea (MNU), and their combinatorial use was studied. Intestinal tumors, including both adenoma and adenocarcinoma, were induced in all the rats given the carcinogens. The tumor frequently occurred in the intestinal tract extending from the duodenum to the colon in the DMH treated group. In the MNU treated group, the tumor occurred in the colon, especially on the distal side. Distribution of tumors in the group treated with both carcinogens was similar to that seen in the MNU-treated group. Neither obvious macroscopic nor histological differences were observed between groups treated with these carcinogens. In addition, we investigated the effect of carcinogens on ornithine decarboxylase (ODC) activity in rat colon. Tumor tissues showed a remarkably high level of enzyme activity compared with the normal-appearing mucosa, and there was a correlation between the level of the ODC activity and the histological grade of malignancy. ODC activity in the normal-appearing mucosa of the carcinogens-treated rats was significantly higher than that of control rats, and the number of tumors per rat was correlated with the level of ODC activity. These results indicate that mucosal ODC may be a pertinent biological marker for local carcinogenic activity.

Age-related changes in polyamine biosynthesis after fasting and refeeding

The effect of aging on ornithine decarboxylase (ODC) activity and polyamine biosynthesis in the proximal small intestine was studied in two groups of male Fisher 344 rats (young [4-month old] and aged [26- to 27-month old]) using a fasting and refeeding model. In control (nonfasted) rats, levels of polyamines (putrescine, spermidine and spermine) and ODC activity were significantly higher in aged compared with young rats. In aged rats, fasting significantly reduced the levels of putrescine by 41%, spermidine by 23%, and spermine by 11%; however, fasting had no effect on polyamine levels in young rats. ODC activity was decreased 75% in young and 50% in aged rats after fasting compared with the respective age-matched controls. Conversely, 2 h after reinstituting a chow diet increased ODC activity by 17-fold in young rats but only 8-fold in aged rats. Putrescine levels were also increased in both age groups after refeeding; however, similar to ODC activity, these increases were much less in aged rats. In addition, spermidine and spermine levels remained significantly depressed in the aged groups even after 24 h of refeeding. These findings suggest that the normal rigid control of gut polyamine biosynthesis and proliferation noted in young rats is markedly altered with aging.

Effects of the Protein Phosphatase Inhibitor Okadaic Acid on the Actions of Prolactin in Cultured Mouse Mammary Gland Explants

The effects of okadaic acid on ornithine decarboxylase (ODC) activity and milk product formation were studied in cultured mammary tissues derived from midpregnant mice. Okadaic acid stimulated ODC activity as well as the synthesis of lactose, lipids, and casein-rich phosphoproteins. When tested together, okadaic acid and prolactin evoked a nonadditive stimulation of lactose synthesis. In addition, the time-courses for the okadaic acid and prolactin effects on lactose synthesis were the same. In contrast to the effects of okadaic acid and prolactin on lactose synthesis, these agents generated additive stimulatory effects on ODC activity and the synthesis of lipids and casein-rich phosphoproteins. Since okadaic acid is known to inhibit protein phosphatases-1 and 2A, we conclude that the substrate(s) of these phosphatases are involved in the signal transduction pathway by which prolactin expresses its effect on lactose synthesis in mammary tissues. Whether these substrates are also involved in other actions of prolactin in mammary tissues remains to be established.

Subdiaphragmatic vagotomy abolishes increase in ornithine decarboxylase activity of rat duodenal mucosa after ischemia-reperfusion in superior mesenteric artery.

In order to evaluate the systemic and neural factors on ornithine decarboxylase (ODC) activity of rat intestinal mucosa, ODC activity in duodenal mucosa, where blood flow did not decrease by superior mesenteric artery (SMA) occlusion, was compared to that of jejunal and ileal mucosa, where blood flow decreased to 90% of the initial value after SMA occlusion. Rats were allowed to recover after SMA occlusion before harvesting intestinal mucosa for measuring ODC activity. ODC activity in the jejunum and the ileum increased markedly 6 h after ischemia-reperfusion (I/R) and more than 72 h were required for ODC activity to return to normal. ODC activity in the duodenum did not change until 24 h after I/R, but the activity increased 48 h after I/R, and this increase continued for 2 days. Subdiaphragmatic vagotomy completely abolished the increase in ODC activity in the duodenum, whereas the same procedure had no influence on ODC activity in the jejunum or the ileum. These results indicate that a neural signal from the central nervous system via the efferent vagal nerve following I/R was an important factor in the increase in ODC activity of duodenal mucosa, where blood flow was not influenced by SMA occlusion.