Changes In Ornithine Decarboxylase Activity Research Articles

EFFECT OF WATER-SOLUBLE QUERCETIN ON INDICATORS OF OXIDATIVE-NITROSATIVE STRESS IN TISSUES OF SUBMANDIBULAR SALIVARY GLANDS OF RATS UNDER LIPOPOLYSACCHARIDE-INDUCED SYSTEMIC INFLAMMATORY RESPONSE

The aim of this study was to investigate the effect of the water-soluble form of quercetin on the indicators of oxidative-nitrosative stress in the tissues of the submandibular salivary glands (SG) under conditions of lipopolysaccharide-induced systemic inflammatory response (SIR). The study was performed on 30 white Wistar male rats weighing 180-220 g, divided into 3 groups: 1st group included intact animals, 2nd group included animals, subjected to the systemic administration of lipopolysaccharide of Salmonella typhi, 3rd group involved animals, which received water-soluble quercetin complex with polyvinylpyrrolidone (corvitin) in a dosage of 100 mg/kg (10 mg/ g in terms of quercetin) intraperitoneally every 3 days, starting on the 30 day of the experiment with using S. typhi lipopolysaccharide. The latter was administered in a dose of 0.4 μg/kg body weight 3 times during the 1st week, and then once a week for the next 7 weeks. The study has demonstrated that applying quercetin under SIR restrains the production of reactive oxygen species in SG tissues: it reduces unstimulated production of superoxide anion radical and its induced generation with the administration of NADPH (by microsomes and NO-synthase), NADH (by mitochondria), S. typhi lipopolysaccharide (by leukocyte NADPH-oxidase). The administration of quercetin under SIR condition diminishes the signs of nitrosative stress in SG tissues, as evidenced by decrease in inducible NO-synthase activity without significant changes in ornithine decarboxylase activity and the level of coupling of constitutive isoform of NO-synthase, decreased concentration of highly active peroxynitrite, but, however, without significant shift in S-nitrosothiols content.

Organophosphorus insecticides affect normal polyamine metabolism in amphibian embryogenesis

The objective of the present study was to evaluate the concentration- and time-dependent effects of the organophosphorus insecticides malathion and azinphos-methyl on polyamine metabolism, and relate them to normal and altered embryonic development of the common toad Rhinella arenarum. Control embryos showed that the higher polyamines spermidine and spermine acquired importance with respect to the diamine putrescine as embryonic development progressed. The activity of ornithine decarboxylase significantly decreased in complete operculum embryos. Continuous exposure to malathion caused a decrease in polyamine levels during embryonic development. However, there was an increase in putrescine levels in complete operculum embryos exposed to a sublethal concentration of the insecticide. Embryos exposed to malathion displayed a decrease in fresh weight and size, along with an increase in the number of malformed individuals. R. arenarum embryos exposed to a lethal concentration of azinphos-methyl showed an increase in putrescine levels and a decrease in spermidine and spermine levels, accompanied by an increase in ornithine decarboxylase activity. In conclusion, as the embryonic development of the toad R. arenarum progresses, polyamine metabolism shifts to higher polyamine levels with a more preponderant contribution of spermidine and spermine with respect to putrescine and involves a dramatic change in ornithine decarboxylase activity, one of the key regulatory enzymes of the pathway. Organophosphorus insecticides are capable of altering polyamine metabolism, slowing embryo development in parallel with a reduction in spermidine and spermine levels. An increase in the oxidative degradation of polyamines might be involved in the toxic action of organophosphorus insecticides and might also be related to other effects such as teratogenesis.

Regulatory effects of corticosterone on ornithine decarboxylase activity during liver regeneration in rats

The regulation of ornithine decarboxylase (ODC) gene expression and enzyme activity by corticosterone during rat liver regeneration induced by partial hepatectomy (PH) was evaluated. Bilateral adrenalectomies were performed on ether-anesthetized rats 3 days before PH. Corticosterone in sesame oil was injected subcutaneously to adrenalectomized rats. ODC mRNA, ODC protein and enzyme activity were detected by in situ hybridization, Western blot and high performance liquid chromatography (HPLC), respectively. The ODC mRNA levels, protein accumulation and enzyme activity were lower in the intact liver compared to the regenerating liver. After PH, mRNA levels were remarkably enhanced in all groups and peaked at 5 h post-PH, and presented a persistent increase only in adrenalectomy rats during the regeneration process. Corticosterone treatment resulted in a dose-dependent decrease in ODC mRNA content after 5 h post-PH. ODC protein accumulation in adrenalectomy rats was higher than that in sham-adrenalectomy rats, but it decreased in corticosterone-treated (10 mg/kg) rats until 24 h post-PH, with a strong decline seen in 40 mg/kg corticosterone-treated rats. ODC activity was rapidly promoted, and the highest levels were observed at 6 h after PH in all groups. After corticosterone treatment, the activities declined significantly at 6 h post-PH, with the lowest value found in the 40 mg/kg group. Corticosterone treatment results in dose-dependent decreases in ODC mRNA and enzyme protein both in the intact liver and the regenerating liver. The change in ODC activity is partially related to alterations of ODC mRNA and protein accumulation.

Changes in ornithine decarboxylase activity in response to temperature stress and stimulation of juvenile hormone in Anastrepha fraterculus (Diptera, Tephritidae)

Ornithine decarboxylase (ODC) activity was analyzed in Anastrepha fraterculus (Diptera) females (4 days old) submitted to temperature stress (6 degrees C and 20/6 degrees C) and the topical application of juvenile hormone (JH). ODC activity and ejaculatory apodeme measurements (length and width) were made in males (15 days old) after 6 degrees C stress. JH dose of 500 ng and incubation of 3, 7, and 18 h increased ODC activity. Females reared at 6 degrees C and 20/6 degrees C had higher ODC activity than those reared at 25 degrees C. The treatment of 6 degrees C and JH incubation for 1 h increased ODC activity when compared to 6 degrees C treatments only. However, the treatment of 20/6 degrees C only after 3 or 18 h of JH incubation resulted in higher ODC activity than controls (20/6 degrees C) or 20/6 degrees C plus 1 h of JH incubation. Males did not undergo differences in ODC activity when reared at 6 degrees C or 25 degrees C but the ejaculatory apodeme measurements was higher in those reared at 25 degrees C than in those reared at 6 degrees C. The results can be considered an adaptive process to environmental changes.

Induction of apoptosis in human leukaemic cells by IPENSpm, a novel polyamine analogue and anti-metabolite.

Human promyelogenous leukaemic cells (HL-60) were treated with novel spermine analogue, ( S )- N (1)-(2-methyl-1-butyl)- N (11)-ethyl-4,8-diazaundecane (IPENSpm), and the effects on growth and intracellular polyamine metabolism were measured. IPENSpm was cytotoxic to these cells at concentrations greater than 2.5 microM. It induced apoptosis in a caspase-dependent manner and its toxicity profile was comparable with etoposide, a well-known anti-tumour agent and inducer of apoptosis. IPENSpm decreased intracellular polyamine content as a result of changes in ornithine decarboxylase activity and increases in spermidine/spermine N(1)-acetyltransferase and polyamine export. Analysis showed spermine and spermidine as the major intracellular polyamines, while putrescine and acetyl-polyamines were the main export compounds. IPENSpm used the polyamine transporter system for uptake and its accumulation in cells was prevented by polyamine transport inhibitors. IPENSpm can be classified as a polyamine anti-metabolite and it may be a promising new lead compound in terms of treatment of some human cancers.

Abnormal ornithine decarboxylase activity in transgenic mice increases tumor formation and infertility

A transgenic mouse line carrying ornithine decarboxylase cDNA as the transgene under the control of a mouse mammary tumor virus long terminal repeat (MMTV LTR) promoter was generated in order to study whether ornithine decarboxylase transgene expression will have any physiological or pathological effect during the entire life of a transgenic mouse. The high frequency of infertile animals and the loss of pups made the breeding of homozygous mice unsuccessful. However, a colony of heterozygous transgenic mice was followed for 2 years. In adult heterozygous transgenic mice, ornithine decarboxylase activity was significantly increased in the testis, seminal vesicle and preputial gland when compared to non-transgenic controls. In contrast, ornithine decarboxylase activity was decreased in the kidney and prostate of transgenic mice. No significant changes in ornithine decarboxylase activity were found in the ovary and mammary gland and only moderate changes in ornithine decarboxylase activity were detected in the heart, brain, pancreas and lung. The most common abnormalities found in adult animals (12 males and 20 females) of the transgenic line were inflammatory processes, including pancreatitis, hepatitis, sialoadenitis and pyelonephritis. Spontaneous tumors were observed in eight animals, including two benign tumors (one dermatofibroma, one liver hemangioma) and six malignant tumors (one lymphoma, one intestinal and three mammary adenocarcinomas and one adenocarcinoma in the lung). No significant pathological changes were found in 17 nontransgenic controls.

Inhibition of 12- O-tetradecanoylphorbol-13-acetate (TPA)-induced mouse skin ornithine decarboxylase and protein kinase C by polyphenolics from grapes

Ornithine decarboxylase is the rate-limiting enzyme in the biosynthesis of polyamines, which are believed to play an essential role in diverse biological processes including cell proliferation and differentiation. We have previously reported [J. Bomser, K. Singletary, M. Wallig, M. Smith, Inhibition of TPA-induced tumor promotion in CD-1 mouse epidermis by a polyphenolic fraction from grape seeds, Cancer Letters 135 (1999) 151-157] that pre-application of a grape polyphenolic fraction (GPF) to mouse skin epidermis inhibits 12- O-tetradecanoylphorbol-13-acetate (TPA)-induced ornithine decarboxylase (ODC) activity, as well as 7,12-dimethylbenz[a]anthracene (DMBA)-initiated, TPA-promoted mouse skin tumorigenesis. The present studies were designed to further characterize the effect of time and dose of application of GPF on TPA-induced ODC activity and protein expression, and on protein kinase C activity in mouse skin epidermis. In addition, the effect of GPF on ODC kinetics in vitro was examined. Application of 5, 10, and 20 mg of GPF 20 min prior to treatment with TPA resulted in a significant decrease in epidermal ODC activity of 54, 53, 90%, respectively, compared with controls. Yet, ODC protein levels (Western blot) in the 10 and 20 mg GPF groups were significantly increased by 1.8 and 1.9-fold, respectively, compared with controls. A similar response was observed with the ODC inhibitor 2-difluoromethylornithine (DFMO), which served as a positive control. Application of grape polyphenolics (20 mg) at 60 and 30 min prior to treatment with TPA inhibited ODC activity by 62 and 68%, respectively, compared with controls ( P<0.05). In contrast, application of grape polyphenolics (20 mg) at 60, 120 and 240 min after treatment with TPA resulted in no significant changes in ODC activity. A similar increase in epidermal ODC protein was observed in these GPF-treated animals, similar to that observed when GPF application preceded TPA. When applied to mouse skin prior to TPA, GPF was associated with a decrease in subsequent PKC activity compared with controls at 10 and 30 min following TPA treatment. The GPF-associated decrease in PKC activity preceded the decrease in ODC activity. In a separate in vitro study, kinetic analyses indicated that GPF is a competitive inhibitor of ODC activity. Collectively these data suggest that the grape polyphenolic fraction is effective as an inhibitor of ODC activity when applied before TPA, and that the magnitude of inhibition is independent of epidermal ODC protein content. In addition, GPF is a competitive inhibitor of ODC activity in vitro. The decrease in TPA-induced ODC activity due to GPF treatment is preceded by an inhibition of TPA-induced PKC activity. Thus, the polyphenolic fraction from grapes warrants further examination as a skin cancer chemopreventive agent that interferes with cellular events associated with TPA promotion.

Increased translation efficiency and antizyme-dependent stabilization of ornithine decarboxylase in amino acid-supplemented human colon adenocarcinoma cells, Caco-2

The mechanisms of the response of ornithine decarboxylase (ODC), the rate-limiting enzyme in polyamine biosynthesis, to amino acid supplementation were studied in the human colon adenocarcinoma cell line, Caco-2. Supplementation of serum-deprived, subconfluent Caco-2 cells with any one of a series of amino acids (10 mM) resulted in increased ODC activity, reaching a maximum of approx. 12.5-fold after approx. 4 h, over control cells either not supplemented or supplemented with iso-osmolar D-mannitol. Glycine, L-asparagine and L-serine, as well as their D-enantiomers, were the strongest effectors and acted in a concentration-dependent manner; millimolar concentrations of most of these amino acids being sufficient to significantly increase ODC activity. In contrast, supplementation with D-methionine, L-lysine, L-aspartate or L-glutamate had little or no effect on ODC activity, whereas supplemental L-methionine, L-arginine, L-ornithine or L-cysteine was inhibitory. Polyamine assays showed that the putrescine content of cells varied in accordance with the changes in ODC activity. Western-blot and Northern-blot analyses revealed specifically increased levels of ODC protein but not mRNA, respectively, in response to supplementation with an ODC-inducing amino acid. Suppression of the increase in cycloheximide-treated cells confirmed a requirement for protein synthesis. Pulse-labelling of cells with [35S]methionine showed a 3-fold increase in the synthesis of ODC protein after 4 h of supplementation with glycine or L-serine. Supplemental glycine also augmented, reversibly, the half-life of ODC by almost 4-fold and simultaneously decreased the activity of putrescine-induced free antizyme. These results suggest that translational, but not transcriptional, regulation of ODC takes part in ODC induction by amino acids in Caco-2 cells. However, it also appears to occur in concert with decreased enzyme inactivation and/or degradation.

Increased translation efficiency and antizyme-dependent stabilization of ornithine decarboxylase in amino acid-supplemented human colon adenocarcinoma cells, Caco-2

The mechanisms of the response of ornithine decarboxylase(ODC), the rate-limiting enzyme in polyamine biosynthesis, to amino acid supplementation were studied in the human colon adenocarcinoma cell line, Caco-2. Supplementation of serum-deprived, subconfluent Caco-2 cells with any one of a series of amino acids (10 mM) resultedin increased ODC activity, reaching a maximum of approx. 12.5-fold after approx. 4 h, over control cells either not supplemented or supplemented with iso-osmolar D-mannitol. Glycine, L-asparagine and L-serine, as well as their D-enantiomers, were the strongest effectors and acted in a concentration-dependent manner; millimolar concentrations of most of these amino acids being sufficient to significantly increase ODC activity. In contrast, supplementation with D-methionine, L-lysine, L-aspartate or L-glutamate had little or no effect on ODC activity, whereas supplemental L-methionine, L-arginine, L-ornithine or L-cysteine was inhibitory. Polyamine assays showed that the putrescine content of cells varied in accordance with the changes in ODC activity. Western-blot and Northern-blot analyses revealed specifically increased levels of ODC protein but not mRNA,respectively, in response to supplementation with an ODC-inducing amino acid. Suppression of the increase in cycloheximide-treated cellsconfirmed a requirement for protein synthesis. Pulse-labelling of cellswith [(35)S]methionine showed a 3-fold increase in thesynthesis of ODC protein after 4 h of supplementation with glycineor L-serine. Supplemental glycine also augmented, reversibly, the half-life of ODC by almost 4-fold and simultaneously decreased the activity of putrescine-induced free antizyme. These results suggest that translational, but not transcriptional, regulation of ODC takes part in ODC induction by amino acids in Caco-2 cells. However, it also appears to occur in concert with decreased enzyme in activation and/or degradation.

Changes in ornithine decarboxylase activity and putrescine concentrations after spinal cord compression injury in the rat

Traumatic spinal cord injury results in direct physical damage to structures and the generation of local factors contributing to secondary pathogenesis. In the present study, we investigated changes in polyamine metabolism after spinal cord compression injury in the rat. This is a stress induced metabolic pathway, of which an activation may indicate both, secondary pathogenesis or induction of neuroprotective response. Ornithine decarboxylase (ODC) activity, the rate limiting step of polyamine synthesis, and levels of the diamine putrescine, the product of ornithine decarboxylase reaction, were analyzed in control (non-laminectomized) animals and at 2 and 4 h after laminectomy or compression injury at the L4 segmental level. ODC activity was significantly increased 4 h after laminectomy in L4 and in adjacent L3 and L5 segments and compression to L4 produced a further increase 4 h after injury as compared with the intact control group. Putrescine levels were likewise significantly elevated to the same extend in the laminectomized and injured cord as compared with the intact control group. These findings demonstrate increased ODC and putrescine levels in the laminectomized and traumatized spinal cord and suggest that laminectomy may be an important `priming event' that contributes to secondary injury after spinal cord compression injury.

Role of modulation on the effect of microwaves on ornithine decarboxylase activity in L929 cells.

The effect of 835 MHz microwaves on the activity of ornithine decarboxylase (ODC) in L929 murine cell was investigated at an SAR of approximately 2.5 W/kg. The results depended upon the type of modulation employed. AM frequencies of 16 Hz and 60 Hz produced a transient increase in ODC activity that reached a peak at 8 h of exposure and returned to control levels after 24 h of exposure. In this case, ODC was increased by a maximum of 90% relative to control levels. A 40% increase in ODC activity was also observed after 8 h of exposure with a typical signal from a TDMA digital cellular telephone operating in the middle of its transmission frequency range (approximately 840 MHz). This signal was burst modulated at 50 Hz, with approximately 30% duty cycle. By contrast, 8 h exposure with 835 MHz microwaves amplitude modulated with speech produced no significant change in ODC activity. Further investigations, with 8 h of exposure to AM microwaves, as a function of modulation frequency, revealed that the response is frequency dependent, decreasing sharply at 6 Hz an 600 Hz. Exposure with 835 MHz microwaves, frequency modulated with a 60 Hz sinusoid, yielded no significant enhancement in ODC activity for exposure times ranging between 2 and 24 h. Similarly, exposure with a typical signal from an AMPS analog cellular telephone, which uses a form of frequency modulation, produced no significant enhancement in ODC activity. Exposure with 835 MHz continuous wave microwaves produced no effects for exposure times between 2 and 24 h, except for a small but statistically significant enhancement in ODC activity after 6 h of exposure. Comparison of these results suggests that effects are much more robust when the modulation causes low-frequency periodic changes in the amplitude of the microwave carrier.

Effect of temporary reductions in feeding on protein synthesis and energy storage of juvenile Atlantic salmon

Biochemical and tissue‐composition indices were used to determine the impacts of temporary feeding reductions on juvenile Atlantic salmon (4.2g initial weight). Three levels of food reduction (fasting, 20% of control, 50% of control) lasting for 2, 4 or 8 days were compared to a control group (constant feeding at 2.75% of body weight day−1). Ornithine decarboxylase (ODC) activity and RNA concentrations were used to measure changes in protein synthesis; condition factor (K), liver‐somatic index (LSI), visceral‐somatic index (VSI), and gut index (GI)were used to provide information on stored energy levels. Results showed that physiological adjustments to decreases or increases in feeding occur very quickly. The most rapid responses were changes in ODC activity (within 2 days) and LSI (2–4 days). Changes in RNA concentrations and K required 4–8 days. There was no apparent effect of the treatments on VSI or GI. Temporary reductions of feeding lasting 2 days or less would not be detected by any index except ODC activity. Changes in the indices are discussed with respect to short‐term changes in rates of protein synthesis and energy storage.

Enhancing effect of ultraviolet A on ornithine decarboxylase induction and dermatitis evoked by 12-o-tetradecanoylphorbol-13-acetate and its inhibition by curcumin in mouse skin.

Previous studies have demonstrated appreciable tumor induction in mouse skin by daily irradiation with high-power long-wavelength ultraviolet A (UVA). The aim of the present study was to examine the enhancing effects of UVA on changes in mouse skin mediated by the tumor promoter 12-o-tetradecanoylphorbol-13-acetate (TPA) by measurement of ornithine decarboxylase (ODC) activity and morphometric analysis. In addition, we examined the inhibitory effects of curcumin, a component of turmeric, on these changes. ODC activity in the epidermis of CD-1 mice was determined by the method of Russell and Snyder. Epidermal and dermal thickness, and the number of dermal infiltrating inflammatory cells were quantified using a computer-assisted image analyzer. A combination of topical TPA application and UVA irradiation produced a greater increment of ODC activity at 4 h than TPA alone (p < 0.05). Histopathologically, TPA plus UVA tended to increase the dermal infiltrating inflammatory cells in contrast to TPA alone. Pretreatment of mice with curcumin significantly abrogated the TPA-induced changes in ODC activity and the dermal infiltrating inflammatory cells as well as the TPA plus UVA-mediated enhancement of these changes. Our data indicate that UVA irradiation (18.72 J/cm2) significantly enhances ODC induction at an early stage (4-6 h) after topical application of TPA, and aggravates the dermatitis elicited by TPA. Pretreatment with curcumin significantly inhibits these enhancing effects.

Relationship between Ornithine Decarboxylase and Cytoskeletal Organization in Cultured Human Keratinocytes: Cellular Responses to Phorbol Esters, Cytochalasins, and α-Difluoromethylornithine

Changes in cell shape occur during the cell cycle and influence cell proliferation and differentiation. In order to study how altered cell proliferation and cell shape are interrelated, we have studied ornithine decarboxylase (ODC) regulation in cultured normal human epidermal keratinocytes (NHEK). Cytoskeletal disruptors have been reported to modulate regulation of ODC; the products of ODC, the polyamines, influence actin polymerization rates in vitro , and polyamine auxotrophs have profoundly disrupted cytoskeletons. Therefore, altered ODC levels could be involved in signaling changes in cell shape and an intact cytoskeleton could transduce signals to regulate ODC levels. We had previously observed that the phorbol ester 12- O-tetradecanoylphorbol-13-acetate (TPA), which profoundly alters cell shape, markedly suppresses ODC biosynthesis in NHEK solely at posttranscriptional/protein synthesis levels. TPA treatment caused NHEK to rapidly assume a rounded morphology that was accompanied by a change in actin organization, as determined by rhodamine-phalloidin labeling. Immunolocalization of ODC showed a perinuclear/nuclear distribution in untreated NHEK and a more diffuse pattern after TPA treatment that was apparent within 15-30 min. Changes in ODC enzyme activity are not significant until 60 min after TPA treatment. NHEK treated with cytochalasin B or D to inhibit actin polymerization exhibited a diffuse ODC localization that could he reversed by removal of the cytochalasin; inhibition of ODC by α-difluoromethylornithine caused a diffuse ODC localization. All treatments resulted in cytoskeletal remodeling. These data are the first evidence for a distinct subcellular localization for ODC and suggest that changes in ODC localization may be an initial step in regulation of ODC activity. Furthermore, changes in ODC activity cause an altered cytoskeleton, suggesting one means by which growth regulatory signals can be transduced to the cytoskeleton from various signaling pathways.

Regulation of ornithine decarboxylase during cell growth. Changes in the stability and translatability of the mRNA, and in the turnover of the protein.

When Ehrlich ascites tumor cells were stimulated to grow, their ornithine decarboxylase (ODC) activity increased 20- to 30-fold. The increase in ODC mRNA content was one order of magnitude less during the corresponding period. Likewise, the subsequent changes in ODC activity failed to show proportionality to those of the ODC mRNA content. The changes in ODC activity were not attributable to changes in ODC turnover, even though the half-life of the enzyme decreased from 56 min during the period of increasing, to 36 min during the period of decreasing ODC activity. There was no evidence of an activation-inactivation-cycle for the enzyme. In view of these findings it appears that ODC mRNA alterations are amplified mainly at the translational level. The biphasic change in ODC mRNA content was partly attributable to a change in turnover of the message, as determined after inhibition of transcription with actinomycin D. Thus, the ODC mRNA half-life was estimated to decrease from 8.7 h during the period of increasing ODC activity to 4.0 h during the period of decreasing ODC activity. Despite the inhibition of transcription by actinomycin D, there was a marked superinduction of ODC activity. Our data demonstrate that the regulation of ODC expression is a complex phenomenon, involving controls at many levels.

Expression of ornithine decarboxylase in pancreatic development.

We examined the relationship between ornithine decarboxylase (ODC) and growth and differentiation in the developing rat exocrine pancreas. The ODC activity profile showed 2 distinct stages of increases with the first occurred at 14-16 days of age, and a second at 21-23 days of age. Growth parameters evaluated as gains in tissue mass, protein and DNA content in the pancreas indicated a low growth rate soon after birth with a transition to a much more rapid growth rate around the age of 20-21 days, a time corresponded to the second rise in ODC activity. Differentiation parameters evaluated as the accumulation of trypsinogen, amylase and lipase showed different temporal changes. While the rate of accumulation of all three enzymes was relatively low following birth, a rapid rate of accumulation of trypsinogen and amylase started around 15-16 days, a time corresponding to the first rise in ODC activity. Lipase, however, did not show an increase in its accumulation until around age 20 days. These results indicate that a rise in ODC activity is closely associated with growth and differentiation in the developing rat pancreas. To further examine this issue, the steady state levels of ODC mRNA in developing rats were evaluated by Northern blots probed with an ODC cDNA. The developmental profile of ODC mRNA showed a broad peak with a pronounced shoulder occurring at 10 days of age. A higher peak was reached around 20 days of age, then dropped precipitously to a very low level at the age of 24 days. This temporal changes in the level of ODC mRNA show good relationship to the changes in ODC activity suggesting that the control of ODC expression occurs at least in part at the pre-translational level.

Post-Transcriptional Suppression of Human Ornithine Decarboxylase Gene Expression by Phorbol Esters in Human Keratinocytes

The induction of ornithine decarboxylase levels by the phorbol ester 12-0-tetradecanoyl-phorbol-13-acetate (TPA) in mouse skin has been shown to be integral to tumor promotion by TPA, and changes in ornithine decarboxylase activity indicate the proliferative state of many different cell types. However, in cultured human epidermal cells, TPA has been reported to be antiproliferative. Therefore, to elucidate pathways that TPA activates in cultured human skin cells, we have examined the levels at which TPA regulates ornithine decarboxylase gene expression in two immortalized human epidermal keratinocyte cell lines, and in normal neonatal keratinocytes. We have found that in cultured human keratinocytes, TPA cases a marked decrease in ornithine decarboxylase enzyme activity (50-90%), with no detectable effect on ornithine decarboxylase mRNA levels. TPA decreased steady-state levels of ornithine decarboxylase immunoreactive protein (approximately 50-67%), accounting for the 50-90% suppression of ornithine decarboxylase activity levels, as well as decreasing new synthesis of ornithine decarboxylase protein (48-50%). However, measurement of ornithine decarboxylase protein half-life showed no significant effect of TPA. Also, prolonged treatment of keratinocytes with phorbol esters abolished the suppression of ornithine decarboxylase activity by TPA. Our data, therefore, suggest that phorbol esters suppress ornithine decarboxylase gene expression predominantly by decreasing ornithine decarboxylase mRNA translatability.

Enhancing Effects of Organosulfur Compounds from Garlic and Onions on Hepatocarcinogenesis in Rats: Association with Increased Cell Proliferation and Elevated Ornithine Decarboxylase Activity

Four organosulfur compounds from garlic and onions were examined for modifying effects on diethylnitrosamine (DEN)‐induced neoplasia of the liver in male F344 rats using the medium‐term bioassay system based on the two‐step model of hepatocarcinogenesis. Carcinogenic potential was scored by comparing the numbers and areas per cm2 of induced glutathlone S‐transfcrasc placental form‐positive foci. Isothiocyanic acid isobutyl ester (IAIE), dipropyl trisulfide (DPT), and allyl mercapton (AM) exerted enhancing effects on their development, while dimethyl trisulfide also tended to increase them. To investigate possible mechanisms of the modifying influence, sequential changes in ornithine decarboxylase activity (ODC) over 24 h were measured in AM‐treated liver tissue without prior DEN initiation. The activity started to increase by 4 h after AM‐treatment, and reached maximum at 16 h, compared to controls. Spermidine/spermine N1‐acetyltransferase activity was not significantly changed. An increase in proliferating cell nuclear antigen‐positive cells followed the elevation of ODC activity. These results suggest that IAIE, DPT, and AM promote rat hepatocarcinogenesis and their promoting effect might be caused by increased cell proliferation with increased poly‐amine biosynthesis. In evaluating relationships between diet and cancer, it is thus appropriate to consider not only a possible protective role of garlic and onions, but also enhancing effects.

Postprandial Changes in Ornithine Decarboxylase Activity, and the Mucosal and Intraluminal Polyamine Levels in the Small Intestine of Rats Concerning the Significance of Intestinal Putrescine Absorption

The dietary response of mucosal ornithine decarboxylase (ODC) in meal-fed and refed rats was investigated for each quarter of the small intestine. The intake of a normal powdered diet caused a considerable postprandial increase in the ODC activity in the mid two quarters, while the intragastric infusion of an aqueous solution containing amino acids and glucose elevated the ODC activity to a large extent in the proximal quarter as well as in its contiguous mid quarter. Such responsiveness was almost completely depressed by previous α-difluoromethylornithine (DFMO) administration. Mucosal scrapings from the small intestine of rats with ODC depression did not significantly differ from those without ODC depression as to their [3H]thymidine-, [3H]uridine-, and [3H]leucine-incorporating capacity into the DNA, RNA, and protein fractions, respectively, at various postprandial times. On the other hand, putrescine was found to plentifully accumulate in the ileal lumen within a few hours after the start of eating, s...

Effect of nitrous oxide on exposure on maternal and embryonic S-adenosylmetthionine levels and ornithine decarboxylase activity

Nitrous oxide is suspected to be a developmental toxicant in humans. The anesthetic does produce increases in the resorption and malformation frequencies in rodents. The mechanism for the drug's developmental toxicant effects in unknown. Embryonic DNA synthesis is decreased; however, this decrease does not appear to be due to depressed levels of adenine or guanine. In this investigation, we examined the effect of N 2O on maternal and embryonic S-adenosylmethionine (AdoMet) levels and ornithine decarboxylase (ODC) activity, and the effect of exogenous methionine (Met) on these parameters was also examined. AdoMet and ODC are involved in polyamine synthesis, and polyamines are involved in regulation of macromolecular synthesis. Pregnant rats were treated with N 2O for 24 hours beginning on the morning of day 10 of gestation. There was no effect of N 2O on maternal hepatic AdoMet or S-adenosylhomocysteine (AdoHcy) levels; there was also no effect on embryonic AdoMet. Embryonic AdoHcy could not be detected in many of the samples; however, N 2O treatment did significantly increase the number of embryonic samples in which AdoHcy was detectable. ODC activity was not affected by either treatment in dams but was increased by N 2O in embryos. It is possible that the embryotoxic effect of this anesthetic is mediated by alterations in the AdoMet to AdoHcy ratio or to changes in ODC activity and polyamine synthesis.