Transcriptomic profile of the porcine oviductal epithelial cells during the peri-conceptional period.

The optimal environment in the oviduct is created by adjusting its ultrastructure and secretory activity to serve the most suitable protection of gametes and to support embryo development. Through gametes/embryo's presence inside the oviduct, the oviductal transcriptomic profile may be altered, and these changes may be caused by DNA methylation. The results of the present study documented that in the epithelial cells of the ampulla and isthmus of the oviducts collected from pigs during the peri-conceptional period, the most differentially expressed genes (DEGs) were down-regulated. Identified DEGs were classified into gene ontology categories as well as annotated into different biological pathways. From evaluated DEGs, genes important for embryo development were selected and the level of DNA methylation was determined. It was documented CLDN18, MUC1, CYP19A3, SOCS1, and ESR1 methylation level have been altered. The presence of embryos in the oviduct changed the transcriptomic profile and the level of DNA methylation in the epithelial cells of ampulla and isthmus during the peri-conceptional period.

Exposure to the electromagnetic field alters the transcriptomic profile in the porcine endometrium during the peri-implantation period.

A low-frequency electromagnetic field (EMF) is an environmental pollutant that may influence female reproduction. This research was undertaken to test the hypothesis that EMF causes alterations in the transcriptomic profile of the endometrium. This study investigated the in vitro effects of EMF treatment (50 Hz, 2 h) on global transcriptome alterations in the endometrium isolated from pigs during the peri-implantation period. The control endometrium was not treated with EMF. The EMF treatment altered the expression of 1561 transcriptionally active regions (TARs) in the endometrium. In the group of 461 evaluated DEGs, 156 were up-regulated (34%), 305 were down-regulated (66%) and 341 (74%) had known biological functions. A total of 210 long noncoding RNAs (lncRNAs) with changes in expression profiles, and 146 predicted RNA editing sites were also evaluated. Exposure to EMF changes the expression of genes encoding proteins that are involved in proliferation and metabolism in endometrial tissue. These results provide useful inputs for further research into the impact of EMF on molecular changes in the uterus during the peri-implantation period and, consequently, pregnancy outcome.

The potential implication of neurokinin B in the modulation of prolactin secretion at the pituitary level in cyclic gilts.

Based on the previous studies, neurokinin B (NKB) participation in the modulation of prolactin secretion at the pituitary level can be assumed, but information concerning this topic is largely insufficient. Therefore, in the present study, we aimed: 1) to evaluate changes in the expression of NKB precursor (Tac3) and its receptor (Tacr3) genes as well as the content of NKB and TACR3 proteins in the porcine anterior pituitary throughout the estrous cycle (days 2 - 3, 9 - 10, 12 - 13, 15 - 16, 19 - 20); 2) to determine in vitro the influence of NKB on the expression of Prl, D2r and Trhr genes in the anterior pituitary cells (incubated for 4 h) as well as on prolactin secretion by these cells (incubated for 4 and 24 h) during chosen days of the estrous cycle (9 - 10, 15 - 16, 19 - 20). The experiments have shown alterations in the expression of Tacr3 mRNA and TACR3 protein content, but not in Tac3 mRNA and NKB protein. The treatment with NKB stimulated the expression of Prl (days 15 - 16), D2r (days 9 - 10) and Trhr (days 19 - 20) genes, but its potential to modulate prolactin secretion was observed only following 24-h incubation, specifically inhibition by NKB alone and stimulation by NKB with dopamine on days 19 - 10 of the cycle. These results indicate some implications of NKB in the modulation of prolactin secretion at the pituitary level in cyclic pigs, however further experiments are required to better clarify its role in this process.

The effect of selective agonists of opioid receptors on in vitro secretion of steroid hormones by porcine endometrium during the estrous cycle and early pregnancy.

Steroid hormones play an important role in the regulation of cyclic changes in the uterus and preparation of intrauterine environment for the egg fertilization, embryo implantation and maintenance of pregnancy. Their secretion by porcine uterus has been demonstrated. The present study aimed to establish the effect of opioid receptors (µ, δ and κ) activation by selective agonists (DAMGO, DPLPE and U 50.488, respectively) on in vitro secretion of steroid hormones (during 6-h and 24-h incubations) by the endometrial explants of gilts on days 2 - 3, 10 - 11, 12 - 3, 15 - 16, 18 - 20 of the estrous cycle and 10 - 11, 12 - 13, 15 - 16 of pregnancy. The agonists at certain of tested concentrations (10-9, 10-8 and 10-7 M) affected secretion of steroid hormones. Progesterone secretion was increased by µ-opioid receptor agonist on days 18 - 20 (6 h) and by δ-agonist on days 2 - 3 and 18 - 20 (24 h) of the cycle. During pregnancy (days 15 - 16), κ-agonist increased it (6 h), but µ-opioid agonist decreased (24 h). Androstenedione secretion was decreased during shorter incubation; by µ- and δ-receptor agonists on days 2 - 3, by all agonists on days 12 - 13, and by κ-receptor agonist on days 18 - 20 of the cycle. However, it was increased during longer incubation with agonists of κ- and µ-opioid receptors on days 10 - 11 and 18 - 20 of the cycle, respectively. Estradiol secretion was elevated by κ- and µ-agonists (6 h) on days 2 - 3 and 15 - 16 of the cycle, respectively, as well as following 24-h incubation with µ-agonist on days 15 - 16, and µ- and κ-agonists on days 18 - 20 of the cycle. During pregnancy, its secretion was increased (24 h) ondays 15 - 16 by µ- and κ-opioid agonists. Cortisol secretion did not significantly change (versus control) in response to applied treatments. These results indicate a potential involvement of EOPs in the modulation of endometrial steroidogenesis in the pig during the estrous cycle and pregnancy.

Effect of neonatal or adult heat acclimation on plasma fT3 level, testicular thyroid receptors expression in male rats and testicular steroidogenesis in vitro in response to triiodothyronine treatment.

This study aimed to evaluate the effect of heat acclimation of neonatal and adult rats on their testes response to in vitro treatment with triiodothyronine (T3). Four groups of rats were housed from birth as: 1) control (CR) at 20°C for 90 days, 2) neonatal heat-acclimated (NHA) at 34°C for 90 days, 3) adult heat-acclimated (AHA) at 20°C for 45 days followed by 45 days at 34°C and 4) de-acclimated (DA) at 34°C for 45 days followed by 45 days at 20°C. Blood plasma and both testes were harvested from 90-day old rats. Testicular slices were then submitted to in vitro treatment with T3 (100 ng/ml) for 8 h. Plasma fT3 level was lower in AHA, NHA and DA groups than in CR group. Basal thyroid hormone receptor α1 (Thra1) expression was higher in testes of NHA and DA and ß1 receptor (Thrb1) in DA rats vs. other groups. In the in vitro experiment, T3: 1) decreased Thra1 expression in all groups and Thrb1 in DA group, 2) increased Star expression in CR, NHA and DA groups, and Hsd17b3 expression in NHA group, 3) decreased the expression of Cyp11a1 in NHA and DA groups, and Cyp19a1 in all the groups, 4) did not affect the activity of steroidogenic enzymes and steroid secretion (A4, T, E2) in all the groups. These results indicate, that heat acclimation of rats, depending on their age, mainly affects the testicular expression of steroidogenic enzymes in response to short-lasting treatment with T3.

Multicenter evaluation of a standardized protocol for noninvasive gene expression profiling.

Gene expression profiling of transplant recipient blood and urine can potentially be used to monitor graft function, but the multitude of protocols in use make sharing data and comparing results from different laboratories difficult. The goal of this study was to evaluate the performance of current methods of RNA isolation, reverse transcription and quantitative polymerase chain reaction (qPCR) and to test whether multiple centers using a standardized protocol can obtain the same results. Samples, reagents and detailed instructions were distributed to six participating sites that performed RNA isolation, reverse transcription and qPCR for 18S, PRF, GZB, IL8, CXCL9 and CXCL10 as instructed. All data were analyzed at a single site. All sites demonstrated proficiency in RNA isolation and qPCR analysis. Gene expression measurements for all targets and samples had correlations >0.938. The coefficient of variation of fold-changes between pairs of samples was less than 40%. All sites were able to accurately quantify a control sample of known concentration within a factor of 1.5. Collectively, we have formulated and validated detailed methods for measuring gene expression in blood and urine that can yield consistent results in multiple laboratories.

The effect of interleukin 1ß and interleukin 6 on estradiol-17ß secretion in the endometrium of pig during early pregnancy and the estrous cycle.

Estrogens are produced by porcine embryos during early pregnancy. It was found that the uterus of pigs might be a source of steroid hormones, including estrogens. However, the factors involved in the regulation of endometrial steroidogenesis remain unknown. We hypothesize that interleukin (IL) 1ß and IL6, which are cytokines produced by porcine embryos and uterine cells, might be involved in the regulation of endometrial estrogen synthesis. Porcine endometrial explants were harvested from gravid (N = 15) and cyclic (N = 15) pigs on days 10 to 11, 12 to 13, and 15 to 16. Samples were analyzed to determine: (1) the expression of CYP19 mRNA and the presence of aromatase cytochrome P450 protein in the tissue; and (2) the release of endometrial estradiol-17ß (E2) in response to IL1ß and IL6 after 6 and 12 hours of in vitro incubation. The effects observed in pregnant gilts were compared with the effects in nonpregnant gilts on corresponding days of the estrous cycle. On days 15 to 16 of pregnancy the expression of CYP19 in the endometrium was markedly decreased when compared with other periods, and the quantity of endometrial P450 aromatase protein was higher on these days than on days 12 to 13. In nongravid pigs, the expression of CYP19 was lower on days 15 to 16 when compared with days 12 to 13 and the quantity of P450 aromatase protein did not differ during the studied days of the estrous cycle. Basal endometrial E2 release was higher in pregnant gilts when compared with cyclic gilts only on days 15 to 16. In gravid pigs IL1ß and IL6 did not affect endometrial E2 release on days 10 to 11 and 12 to 13 of pregnancy (P > 0.05); however, increased E2 release was observed on days 15 to 16 (P < 0.05). In cyclic pigs neither IL1ß, nor IL6 affected endometrial E2 release (P > 0.05). These results provide evidence that: (1) the endometrium possesses a potential for steroidogenesis and produces E2in vitro in gravid and nongravid pigs between days 10 to 16; and (2) IL1ß and IL6 increase in vitro endometrial synthesis of E2 in pregnant pigs on days 15 to 16. Therefore, IL1ß and IL6 might act as stimulators of endometrial E2 secretion in vitro during the time of decreased production of embryonic estrogens. This correlates with a rapid remodeling of the endometrial tissue and the beginning of hemotrophic nutrition of the embryos occurring on days 15 to 16 of pregnancy.

Inhibition of ATM blocks the etoposide-induced DNA damage response and apoptosis of resting human T cells.

It is believed that normal cells with an unaffected DNA damage response (DDR) and DNA damage repair machinery, could be less prone to DNA damaging treatment than cancer cells. However, the anticancer drug, etoposide, which is a topoisomerase II inhibitor, can generate DNA double strand breaks affecting not only replication but also transcription and therefore can induce DNA damage in non-replicating cells. Indeed, we showed that etoposide could influence transcription and was able to activate DDR in resting human T cells by inducing phosphorylation of ATM and its substrates, H2AX and p53. This led to activation of PUMA, caspases and to apoptotic cell death. Lymphoblastoid leukemic Jurkat cells, as cycling cells, were more sensitive to etoposide considering the level of DNA damage, DDR and apoptosis. Next, we used ATM inhibitor, KU 55933, which has been shown previously to be a radio/chemo-sensitizing agent. Pretreatment of resting T cells with KU 55933 blocked phosphorylation of ATM, H2AX and p53, which, in turn, prevented PUMA expression, caspase activation and apoptosis. On the other hand, KU 55933 incremented apoptosis of Jurkat cells. However, etoposide-induced DNA damage in resting T cells was not influenced by KU 55933 as revealed by the FADU assay. Altogether our results show that KU 55933 blocks DDR and apoptosis induced by etoposide in normal resting T cells, but increased cytotoxic effect on proliferating leukemic Jurkat cells. We discuss the possible beneficial and adverse effects of drugs affecting the DDR in cancer cells that are currently in preclinical anticancer trials.

The effect of tumor necrosis factor α (TNFα), interleukin 1ß (IL1ß) and interleukin 6 (IL6) on endometrial PGF2α synthesis, metabolism and release in early-pregnant pigs.

Cytokines produced by the porcine uterus and embryos may be involved in the regulation of endometrial prostaglandin synthesis, metabolism, and release. We studied the effect of tumor necrosis factor α (TNFα), interleukin 1ß (IL1ß) and interleukin 6 (IL6) on: 1) endometrial release of prostaglandin F2α (PGF2α), 2) expression of the terminal enzyme of PGF2α synthesis--PGF synthase mRNA (PGFS mRNA), 3) secretion of PGF(2)α metabolite--13,14-dihydro-15-keto PGF2α (PGFM) by the endometrium and 4) presence and activity of endometrial NAD-dependent 15-hydroxyprostaglandin dehydrogenase (15-PGDH). The effects of cytokines were determined on days 10-11 and days 12-13, e.g., before and during maternal recognition of pregnancy, and on days 15-16, e.g., during the peri-implantation period and compared with its effect in cyclic gilts on corresponding days of the estrous cycle. TNFα did not affect endometrial release of PGF2α in pregnant and cyclic pigs. IL1ß enhanced endometrial PGF2α release on days 12-13 and 15-16 in pregnant and cyclic pigs, respectively. IL6 increased PGF2α release mainly on days 15-16 of pregnancy. Expression of PGFS mRNA was decreased by IL1ß on days 12-13 of pregnancy (P<0.05) and increased in response to IL1ß, TNFα and IL6 on 12-13 (P<0.05) and 15-16 (P<0.01) of the estrous cycle. IL1ß increased release of PGFM in gravid pigs on days 12-13, 15-16 and in non-gravid pigs 10-11 and 15-16 of the cycle. On days 15-16 of pregnancy TNFα and IL6 increased endometrial secretion of PGFM. We determined that in porcine endometrium NAD-dependent 15-hydroxyprostaglandin dehydrogenase (15-PGDH) is present. In gravid pigs, the highest expression of endometrial 15-PGDH occurred during days 12-13 of pregnancy, while in non-gravid pigs during days 10-11 of the estrous cycle. These data provide new evidence that TNFα, IL1ß, IL6 are involved in the regulation of endometrial synthesis, release and metabolism of PGF2α to protect CL during early pregnancy or to facilitate its regression in cyclic females.

In vitro effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on ovarian, pituitary, and pineal function in pigs.

The aims of the study were: (1) to examine 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and/or prolactin (PRL) effects on in vitro secretion of progesterone (P(4)) and estradiol (E(2)) by luteinized granulosa and theca cells from porcine preovulatory follicles; and (2) to determine the effects of TCDD on PRL, luteinizing hormone (LH), and melatonin luteal phase in pigs. We found that TCDD itself did not affect progesterone secretion, but it abolished the stimulatory effect of PRL in the follicular cells. TCDD stimulated PRL secretion during the luteal phase and inhibited during the follicular phase. Moreover, TCDD increased luteinizing hormone secretion by pituitary cells during the follicular phase. In contrast to protein and steroid hormones, melatonin secretion in vitro was not affected by TCDD. In conclusion, it was found that the pituitary-ovarian axis in pigs is sensitive to TCDD, and the dioxin exhibited a profound ability to disrupt the ovarian actions of prolactin.

Curcumin disrupts meiotic and mitotic divisions via spindle impairment and inhibition of CDK1 activity.

OBJECTIVES: Curcumin, a natural compound, is a potent anti-cancer agent, which inhibits cell division and/or induces cell death. It is believed that normal cells are less sensitive to curcumin than malignant cells; however, the mechanism(s) responsible for curcumin's effect on normal cells are poorly understood. The aim of this study was to verify the hypothesis that curcumin affects normal cell division by influencing microtubule stability, using mouse oocyte and early embryo model systems. MATERIALS AND METHODS: Maturating mouse oocytes and two-cell embryos were treated with different concentrations of curcumin (10-50 microm), and meiotic resumption and mitotic cleavage were analysed. Spindle and chromatin structure were visualized using confocal microscopy. In addition, acetylation and in vitro polymerization of tubulin, in the presence of curcumin, were investigated and the damage to double-stranded DNA was studied using gammaH2A.X. CDK1 activity was measured. RESULTS AND CONCLUSIONS: We have shown for the first time, that curcumin, in a dose-dependent manner, delays and partially inhibits meiotic resumption of oocytes and inhibits meiotic and mitotic divisions by causing disruption of spindle structure and does not induce DNA damage. Our analysis indicated that curcumin affects CDK1 kinase activity but does not directly affect microtubule polymerization and tubulin acetylation. As our study showed that curcumin impairs generative and somatic cell division, its future clinical use or of its derivatives with improved bioavailability after oral administration, should take into consideration the possibility of extensive side-effects on normal cells.

The promise of slow down ageing may come from curcumin.

No genes exist that have been selected to promote aging. The evolutionary theory of aging tells us that there is a trade-off between body maintenance and investment in reproduction. It is commonly acceptable that the ageing process is driven by the lifelong accumulation of molecular damages mainly due to reactive oxygen species (ROS) produced by mitochondria as well as random errors in DNA replication. Although ageing itself is not a disease, numerous diseases are age-related, such as cancer, Alzheimer's disease, atherosclerosis, metabolic disorders and others, likely caused by low grade inflammation driven by oxygen stress and manifested by increased level of pro-inflammatory cytokines such as IL-1, IL-6 and TNF-alpha, encoded by genes activated by the transcription factor NF-kappaB. It is believed that ageing is plastic and can be slowed down by caloric restriction as well as by some nutraceuticals. As the low grade inflammatory process is believed substantially to contribute to ageing, slowing ageing and postponing the onset of age-related diseases may be achieved by blocking the NF-kappaB-dependent inflammation. In this review we consider the possibility of the natural spice curcumin, a powerful antioxidant, anti-inflammatory agent and efficient inhibitor of NF-kappaB and the mTOR signaling pathway which overlaps that of NF-kappaB, to slow down ageing.

Interleukin 1ß-induced synthesis and secretion of prostaglandin E2 in the porcine uterus during various periods of pregnancy and the estrous cycle.

Peri-implantation porcine embryos express interleukin-1ß (IL-1ß), which could affect uterine activity during early pregnancy. In vitro studies were conducted to determine if IL-1ß stimulates secretion of PGE2 and expression of cyclooxygenase-2 (COX-2) mRNA and microsomal PGE synthase-1 (mPGES-1) mRNA in uterine tissues harvested from pigs on days 10 to 11, 12 to 13 and 15 to 16 of pregnancy and the estrous cycle. IL-1ß (10 ng/ml) increased PGE2 secretion and mPGES-1 mRNA expression in uterine tissues isolated from pigs between days 10 to 13 of pregnancy and the estrous cycle. IL-1ß stimulated PGE2 and mPGES-1 mRNA expression only in cyclic uterine tissues on days 15 to 16. Interleukin-1ß increased COX-2 mRNA expression in the endometrial tissues of pregnant and cyclic pigs harvested on days 10 to 13. It stimulated COX-2 expression in pregnant pigs' myometrial tissues on days 10 to 11, and on days 15 to 16 in tissues from both pregnant and cyclic pigs. The uterine secretion of PGE2 in response to IL-1ß was determined by local intrauterine concentrations of P4 and E2. This study demonstrates that IL-1ß activates expression of mPGES-1 mRNA in uterine tissues to stimulate synthesis and secretion of PGE2 on days 10 to 13 of both pregnancy and the estrous cycle. The profile of COX-2 mRNA expression in the myometrium differs from its profile in the endometrium. This work provides new insight on the role of IL-1ß in PGE2 production to overcome luteolysis in pregnant pigs.

Effect of aging on UVC-induced apoptosis of rat splenocytes.

UVC-induced apoptotic symptoms such as morphological changes, DNA fragmentation, Bcl-2 and Bax protein expression were examined in primary splenocyte cultures from young (3 months) and old (24 months) rats. The activities of AP-1 and CRE transcription factors in UVC-irradiated splenocytes were also assessed. At 24 h after UVC irradiation 40% of cells derived from young rats were found to be apoptotic, which was twice as much as in splenocytes from old rats. Apoptosis in cells from old rats did not give typical symptoms like a "DNA ladder" and Bcl-2 protein downregulation, in contrast to splenocytes from young rats. No AP-1 transcription factor activity was found in UVC-irradiated splenocytes from old animals and only a trace activity in splenocytes from young animals. This indicates that, UVC-induced apoptosis in rat splenocytes is practically AP-1 independent and that cells from old rats are less sensitive to UVC irradiation than splenocytes from young rats.

Effect of curcumin on the apoptosis of rodent and human nonproliferating and proliferating lymphoid cells.

Curcumin, a major active component of turmeric, has been recognized as an anticarcinogenic agent because of its propensity to induce apoptosis in vivo and in vitro. Previously, we showed that curcumin protects cells against oligonucleosomal DNA fragmentation and induces a novel apoptosis-like pathway in Jurkat cells (Piwocka et al. Exp Cell Res 249, 299-307, 1999). Here, we have studied the ability of curcumin to induce cell death in other human and rodent transformed as well as normal cells. Normal cells were quiescent or stimulated to proliferate. We showed that 50 microM pigment is able to induce cell death in all studied cells, but cell death symptoms varied for different cells. All the cells died as assessed by the TdT-mediated UTP nick end labeling method or trypan blue exclusion test. No one type of cells showed oligonucleosomal DNA fragmentation (DNA "ladder") due to curcumin action, although in HL-60 cells, we were able to observe sub-G1 formation and caspase-3 activation. Together, these data showed that curcumin induces cell death in all tested cells that can be classified as apoptosis-like, and only in HL-60 cells can it be recognized as classical apoptosis.

Glutathione-independent mechanism of apoptosis inhibition by curcumin in rat thymocytes.

Curcumin (CUR) is a natural yellow dye with antioxidant and scavenging properties present in Curcuma species. It is widely used as an anti-inflammatory, anti-mutagenic and chemopreventive agent. In addition to its inhibitory effect on proliferation, CUR has recently been shown to block dexamethasone-induced programmed cell death (apoptosis) of rat thymocytes. Because cellular thiols seem to play a role in redox regulation of apoptosis, the mechanism of the anti-apoptotic effect of CUR was studied by examining the levels of glutathione and acid-soluble sulfhydryl groups. CUR was shown to prevent the glutathione loss occurring in dexamethasone-treated thymocytes, enhancing intracellular glutathione content at 8 hr to 192% of that of nontreated cells. A 60% increase in acid-soluble sulfhydryl groups was also observed. In the presence of L-buthionine S,R-sulfoximine (BSO, an inhibitor of glutathione synthesis), intracellular glutathione content of thymocytes treated with dexamethasone and CUR fell to 31% and that of the acid-soluble sulfhydryl groups to 23% of control after 8 hr. Unexpectedly, the electrophoretic and flow cytometric studies of DNA fragmentation demonstrated that apoptosis did not occur even after 20 hr of incubation with buthionine S,R-sulfoximine and dexamethasone, while control thymocytes and the cells treated only with buthionine S,R-sulfoximine showed DNA fragmentation at a level corresponding to spontaneous apoptosis. These results show that CUR treatment elevated the concentrations of glutathione and nonprotein sulfhydryl groups, thus preventing their decrease in apoptotic thymocytes. Coadministration of L-buthionine S,R-sulfoximine and CUR did not affect the anti-apoptotic effect of CUR suggesting a glutathione-independent mechanism of cell protection.

Inhibition of proliferation and apoptosis of human and rat T lymphocytes by curcumin, a curry pigment.

Curcumin (diferuoylmethane), the yellow pigment in the rhizome of tumeric (Curcuma longa), an ingredient of curry spice, is known to exhibit a variety of pharmacological effects including antitumor, antiinflammatory, and antiinfectious activities. Although its precise mode of action remains elusive, curcumin has been shown to suppress the activity of the AP-1 transcription factor in cells stimulated to proliferate. In this study, we observed that curcumin (50 microM) inhibited proliferation of rat thymocytes stimulated with concanavalin A (Con A) as well as that of human Jurkat lymphoblastoid cells in the logarithmic growth phase. The pigment also inhibited apoptosis in dexamethasone-treated rat thymocytes and in UV-irradiated Jurkat cells as judged by DNA ladder formation, cellular morphological changes, and flow cytometry analysis. The inhibition of apoptosis by curcumin in rat thymocytes was accompanied by partial suppression of AP-1 activity. Complete suppression of AP-1 activity was observed in Con A-treated, proliferating thymocytes. The capacity of curcumin to inhibit both cell growth and death strongly implies that these two biological processes share a common pathway at some point and that curcumin affects a common step, presumably involving a modulation of the AP-1 transcription factor.