T cell stimulator cells expressing find more membrane-bound anti-CD3 antibodies at a high density induced moderate proliferation in human T cells even in the absence of human costimulatory molecules and as expected T cells activated with stimulator cells harbouring high levels of anti-CD3 in combination with human CD80 showed the highest proliferative response (Fig. 1C). To visualize the interaction of human T cells and stimulator cells, we performed co-culture experiments using CFSE-labeled T cells and CMTMR-labeled stimulator cells. Large clusters of T cells and stimulator cells expressing

CD80 can be observed whereas much smaller clusters are formed when T cells were activated by stimulator cells expressing anti-CD3 but no human costimulatory molecule

(Fig. 1D). T cell stimulator cells transduced to express different costimulatory molecules are excellent tools to compare these ligands regarding Epigenetics inhibitor their capacity to activate human T cells. We have generated stimulator cell lines retrovirally expressing different costimulatory molecules at high levels (Fig. 2). The resultant cell lines were used to stimulate purified T cells isolated from different healthy donors and T cell proliferation was assessed. As shown in Fig. 2B stimulation of human T cells in the presence of the costimulatory molecules used in this study (CD80, ICOSL, CD58, CD54 and 4-1BBL) significantly enhanced T cell proliferation compared to T cells co-cultured with stimulator cells expressing no human costimulatory molecule. Furthermore, our data show that CD80 was

the strongest costimulatory ligand tested in these experiments and demonstrate that among the other molecules analyzed CD58 is the most potent inducer of T cell proliferation. There is an increasing number of immunosuppressive and immunomodulatory drugs for treatment of patients suffering from autoimmune diseases and recipients of hematopoietic stem cells or solid organs. Many of these drugs target fast dividing cells whereas others specifically suppress T cells or counteract inflammatory processes. Antibodies or receptor fusion proteins that block the cytokine TNF-α are successfully used in patients suffering from psoriasis, rheumatoid Methisazone arthritis and various other autoimmune diseases (Aringer and Smolen, 2008, Bosani et al., 2009 and Taylor and Feldmann, 2009). TNF-α is a pleiotrophic cytokine and the beneficial effects of TNF-α blockade are mainly ascribed to its capacity to prevent and down-modulate proinflammatory processes. Whereas other members of the TNF-family have been shown to act as potent costimulatory molecules, few studies have addressed the ability of TNF-α to directly contribute to T cell activation processes. We found that expressing TNF-α on T cell stimulator cells enhances their ability to induce proliferation in purified human T cells (Fig. 3A).

Western blots were quantified by NIH ImageJ software version 1.45. Human DKK1 levels in the conditioned medium from HPSE-low and HPSE-high CAG cells were measured using hDKK1 Quantikine ELISA kit (R&D Systems). Mouse DKK1 levels

in conditioned medium from primary murine osteoblastic progenitor cells, C3H10T1/2 pre-osteoblastic cells and ST2 stromal Selleckchem Target Selective Inhibitor Library cells cultured in the absence or presence of rHPSE were measured by mDKK1 Quantikine ELISA (R&D Systems). All assays were performed according to the protocols provided by the manufacturers. Statistical comparisons between two experimental groups were analyzed by Student’s t test. ANOVA was employed for statistical analyses among multiple groups, followed by a post-hoc Bonferroni test. The correlations between heparanase and osteocalcin expression in MM patients’ samples were assessed using Spearman correlation coefficient. P < 0.05 was considered statistically significant and is reported as such. To determine the effects of heparanase expression by myeloma cells on mesenchymal lineage cells, we first

examined the effect of heparanase on osteoblastogenesis and parameters of bone formation by evaluating osteoblast parameters in SCID-hu mice [36]. We measured osteocalcin expression, a marker of mature osteoblast differentiation, in histologic sections of engrafted bone [23]. The analysis revealed that the number of osteocalcin-positive Small Molecule Compound Library osteoblasts was significantly diminished in both primary (injected with tumor cells) and contralateral (not injected with tumor cells) engrafted bones of the mice bearing

HPSE-high tumor, compared to animals bearing tumors formed by HPSE-low cells (Figs. 1A–B). These data provided the first direct experimental evidence that osteoblastogenesis was inhibited by heparanase in vivo. Interestingly, immunohistochemical staining for human kappa light chain, a specific marker of 4-Aminobutyrate aminotransferase CAG myeloma cells, revealed no detectable tumor cells in uninjected contralateral grafts (data not shown), suggesting that the inhibition of osteoblastogenesis was humoral and not due to myeloma tumor metastasis to the contralateral graft. We next investigated the correlation between heparanase expression and osteocalcin expression in myeloma patients, using primary bone marrow core biopsy specimens obtained from myeloma patients. Specific immunohistochemical staining for heparanase and osteocalcin was performed on 28 myeloma patient bone marrow core biopsy specimens. We identified a significant negative correlation (r = − 0.62, P < 0.001) between heparanase expression by myeloma cells and osteocalcin expression by bone marrow cells (Fig. 2 and Supplementary Table 1).

The mean squared error of rˆ is equivalent to 1/η  2. Assuming the normal distribution for rˆk, each σk   is approximated as ¼ of the 95% confidence interval of rˆk (in brackets below) by: equation(3) σk(rˆk,Nk)=14tanhz+1.96Nk-3-tanhz-1.96Nk-3,where equation(4) z=12log1+rˆk1-rˆk,( this website David, 1938) and Nk   is the number of degrees of freedom for a time series of length n  , reduced by the band pass filter to: equation(5) Nk=2ΔTTc1-ΔTTc2(nk-2),( Yan et al., 2004) where ΔTΔT is the time step and Tc  1 and Tc  2 are the band pass times (40 and 160 h, respectively). Although there is autocorrelation in the time series, subsampling at the decorrelation

time causes a negligible change in N  . Each σk  , η  2, rˆk, and rˆ is a function of l, the lead or lag time

between τ and SST. For model correlation Rˆ (model terms represented by capital letters), Eq. (2) reduces because there is a single complete time series, i.e. K   = 1. When comparing between observations and model, the greatest magnitude of the lagged observed and modeled correlation ( rˆ and Rˆ respectively) is selected and denoted r and R, and their associated lead times l and L. For all buoys, r is negative ( Figs. 3 and 4) meaning that increased wind stress leads decreased SST. Lead selleck kinase inhibitor time l   has an observational uncertainty ηl2, calculated by an application of Eq. (2) to lead time where equation(6) ηl2=∑k=1k1σlk2,and the standard deviation for lead time, σl  , has to be estimated empirically. In order to examine σl   for the buoy observations, each lead time l   associated with an individual time series k   (i.e. the time at which rˆk is greatest in magnitude) is subtracted from the mean l   at buoys along its longitude. Shorter time series tend to result in higher deviations

from meridional mean of l  , and the relationship between time series length and lead time variability is even more clear when analyzing artificially truncated model runs ( Fig. 5). Assuming that the record length and standard http://www.selleck.co.jp/products/erastin.html deviation relationship from the observational data is approximated by the model, an exponential fit to the model relationship between standard deviation in l   and record length ( Fig. 5) is used as an approximation of σl   for lead time uncertainty ηl2 (Eq. (6)). Because all model time series have a record length of 2 years, the standard deviation of the estimated error in model lead time, σL, is a constant 2.16 h ( Fig. 5). The uncertainty in forcing is estimated by the sensitivity of model to the blended wind product at each buoy, using the twenty experiments with different wind products and the same model physics (Table 2): equation(7) φ2=1n∑i=120(Ri-μR)2,φ2=1n∑i=120(Li-μL)2,where each i is an experiment forced with a different blended wind, and μ is a mean value over years 1.5–3.5 of the 20 blended wind experiments.

The impacts of normal operations cannot be eliminated, but they can be managed in space and time to minimize effects on culture and environment. Accidents, however, have the potential to cause the most widespread impacts of any of the threats posed by shipping. The record from the nearby Aleutian Islands [77] suggests that over time one or more spills may be close to inevitable. Increasing tug, salvage and spill response capabilities

in the Bering Strait and click here Northwest Arctic should be considered, especially during peak vessel traffic periods. Such capacity could also aid in search and rescue if needed. Local training in emergency response could also selleck chemical enhance the region׳s ability to respond promptly while other assets are en route. Identifying risks and associated regulatory measures is a first step, but taking action will depend also on effective governance of vessel traffic at local, national, and international levels. Bering Strait region communities

will need to develop the technical and human capacity to work effectively with mariners and regulators, to identify community needs and priorities and to implement measures such as local use of AIS and communication systems. National governments will need to continue to develop appropriate regulatory frameworks, including local outreach and involvement as well as standards that are consistent with other such efforts in Arctic waters. Internationally, cooperation between the U.S. and Russia would be a big step forward and would pave the way for recognition of Osimertinib molecular weight appropriate measures by the IMO. In this light, Table 2 outlines the progression from voluntary recommendations to domestic and international regulations. While voluntary recommendations may not be enforceable, they can also be made more quickly than formal regulations, compliance may be high, and they are a significant step towards formal regulations. Formal regulations are likely to take longer to develop and implement, but carry extra

weight. Both approaches have a role in a system of effective governance for vessel traffic. In summary, vessel traffic in the Bering Strait region is an economic opportunity, and also an opportunity for sound management of environmental and cultural risks. This paper presents a framework for various actions that can be taken locally, nationally, and internationally to reduce risks from vessel traffic, consistent with the principle of freedom of the seas as well as with responsible standards of care for vessel operations in areas. Acknowledging the risks and taking appropriate action proactively can help vessel traffic proceed without hindrance, while also protecting an important ecosystem and the cultures that depend on it, while both remain vibrant and healthy.

Several studies in rodent osteoblastic cell lines and bone marrow progenitor cells demonstrated that pharmacological Dapagliflozin AMPK activators metformin and AICAR (acadesine) induce differentiation and mineralization of osteoblasts

by upregulating the expression of Runx2 [25], [26], [27] and [28]. Moreover, the in vivo studies confirmed that metformin stimulates bone lesion regeneration in rats [29], while AMPK gene knockdown reduces bone mass in mice [30] and [31]. Recently, Kim et al. [15], using an RNA interference approach, provided the first evidence for the involvement of AMPK in osteogenic differentiation of human adipose tissue-derived MSC. The results of the present study confirm and expand these findings by demonstrating the induction of autophagy and activation

of Akt as the major early and late downstream events, respectively, in AMPK-controlled MSC osteogenic differentiation. While it has been reported that Akt is required for BMP2-stimulated osteogenesis Anti-infection Compound Library nmr in mice [14] and [32], our data for the first time demonstrate the involvement of autophagy in osteoblast differentiation. The role of AMPK in autophagy induction or Akt activation in osteoblasts has not been assessed thus far, but the present results are consistent with the ability of AMPK to induce autophagy in various cell types [33], as well as to activate Akt in leukemic cells, endothelial cells and renal tubular cells [34], [35] and [36]. The latter effect, however, seems to be cell type- and/or context-dependent, as we have previously failed to observe any influence of AMPK on Akt phosphorylation in U251 human glioma cells exposed to simvastatin or compound C [37] and [38], or in metformin-treated B16 mouse melanoma cell line [39]. While our data with AMPK shRNA clearly support the role of AMPK in Akt activation during osteogenic differentiation of hDP-MSC, it should be noted that the AMPK inhibitor compound C [40] has recently been reported to directly interfere with Akt phosphorylation in an AMPK-independent manner [38]. Therefore, although we used compound C at

quite a low dose (1 μM) as a precaution against non-specific effects, the possibility Roflumilast that its actions in the present study were partly mediated independently of AMPK inhibition could not be completely excluded. However, compound C, unlike Akt inhibitor DEBC, failed to reduce osteogenic differentiation of hDP-MSC if added 3 days after its initiation, which argues against the ability of compound C to directly inhibit Akt in our experimental setting. In addition, it has been shown that AMPK can modulate differentiation of rodent osteoblast cell lines through interference with Wnt/β-catenin and Smad1/5/8-Dlx5 signaling pathways [26] and [41]. We are currently investigating possible connections between these signaling pathways and AMPK-triggered activation of autophagy and Akt during osteoblast differentiation of human MSC.

Die Belege für diese Annahme sind begrenzt. Warfvinge [9] hat jedoch gezeigt, dass die Verteilung von Hg2+ im Cerebellum nach einer Quecksilberdampf-Exposition der Verteilung ähnlich ist, die man nach einer Exposition find more gegenüber Methylquecksilber (MeHg) findet. Da sich die neurotoxischen Effekte von Quecksilberdampf und MeHg stark unterscheiden, schlagen wir dagegen vor, dass MeHg selbst und nicht Hg2+ nach Exposition gegenüber MeHg das letztendlich toxische Agens ist (siehe die ausführliche

Diskussion weiter unten). Dentalamalgam wird seit mehr als 150 Jahren für Zahnfüllungen verwendet. Das Amalgam besteht zu etwa 50% aus metallischem Quecksilber, dazu kommen Silber und Kupfer sowie kleine Mengen anderer Metalle wie z. B. Zink. Die pulverisierten Metalle werden kurz vor der Verwendung mit dem Quecksilber gemischt. Dieser Schritt wurde üblicherweise von Hand durchgeführt, so dass das zahnmedizinische Personal dem Quecksilber ausgesetzt war. Der Einsatz von Dentalamalgam führt also zur Exposition sowohl des zahnmedizinischen Personals als auch der Patienten mit Amalgamfüllungen, da das Quecksilber mit der Zeit aus der Füllung freigesetzt wird. Letzteres kann einem Übersichtsartikel LGK 974 der WHO zufolge die Quelle für eine erhebliche Quecksilberexposition darstellen [10]. Das Ausmaß der Quecksilberfreisetzung aus Füllungen wird vor allem durch den Kauvorgang und die Temperatur

von Speisen bestimmt, wie z. B. im Zusammenhang mit der

Anwendung von Nikotinkaugummi demonstriert Tryptophan synthase wurde [11]. Weiterhin wurde gezeigt, dass der Quecksilbergehalt im Urin die Zahl der Amalgamfüllungen widerspiegelt [12]. Amalgamfüllungen können allergische Reaktionen in der Mundhöhle auslösen, was allerdings sehr selten vorkommt. Davon abgesehen sind die durch Amalgamfüllungen verursachten biologischen Effekte denen von Quecksilberdampf oder Hg2+ ähnlich. In der Literatur gibt es zahlreiche Berichte über Patienten, die angaben, bei sich verschiedene Symptome zu bemerken, welche mit den Symptomen einer Quecksilberdampf-Exposition vergleichbar waren. In einigen dieser Fallberichte wurde außerdem angegeben, dass sich die Symptome nach Entfernen der Amalgamfüllungen besserten. Solche Studien sind nicht einfach durchzuführen, und die Validierung der Ergebnisse ist sogar noch schwieriger. Des Weiteren wurde vermutet, dass Quecksilber Morbus Alzheimer auslösen könnte [13], da Gehirne von Alzheimer-Patienten einen erhöhten Quecksilbergehalt aufwiesen [14]. Dies kann jedoch auch das Ergebnis von Membranschäden sein, die dazu führen, dass die betroffenen Zellen mehr Quecksilber akkumulieren als normale Zellen. So kann man nur spekulieren, was Ursache und was Wirkung ist. Darüber hinaus ergab sich in einer epidemiologischen Studie keine Korrelation zwischen Zahnfüllungen und Alzheimer-Krankheit [15].

2A and 2B) as reported [7] and [12], suggesting specificity of reagent (antibody) and demonstrating a major difference in the levels of BPDE-DNA adducts between exposed and non-exposed animals/tissues. Levels of BPDE-DNA adducts were measured in a similar area of tissue sections (mm2) and

number of cells (∼800 cells/section/animal) in terms of total adduct intensity as well as nuclei containing a percentage of high, medium and low intensity CDK inhibitor due to BPDE-DNA adducts. It was observed that with passage of time, mice on the control diet for 24, 72 and 120 h [subgroups BP(+48h), BP(+96h), BP(+144h)] showed a time-related significant decrease in total adduct(s) intensity (levels) in the liver and lungs compared to BP(+24h) and subgroup of preceding time point (Figure 2 and Figure 3). Interestingly, mice that were shifted to 0.05% curcumin diet and killed at 24, 72 and 120 h [subgroups BP(+48h) + C 24 h, BP(+96h) + C KU-60019 concentration 72 h, BP(+144h) + C 120 h] showed significantly higher decrease in the levels of adducts (intensity) in the liver and lungs compared to BP(+24h) and respective time-matched controls [subgroups BP(+48h), BP(+96h), BP(+144h)] (Figure 2 and Figure 3). This decrease was also evident when a comparison

of percentage intensity of nuclei containing high, medium and low Ribonucleotide reductase levels of adducts was made between curcumin-treated and respective time-matched controls. In the liver, the observed decrease in total adduct intensity in B(a)P [BP(+48h), BP(+96h), BP(+144h)] and B(a)P + curcumin [BP(+48h) + C 24 h, BP(+96h) + C 72 h, BP(+144h) + C 120 h]-treated subgroups

appears to be attributed to the reduction in percentage intensity of nuclei containing high and medium levels of adducts. In the lungs, it was due to decrease in nuclei containing high levels of adducts both in B(a)P [BP(+48h), BP(+96h), BP(+144h)] and B(a)P + curcumin [BP(+48h) + C 24 h, BP(+96h) + C 72 h, BP(+144h) + C 120 h]-treated subgroups (Figs. 2A and 2B). Notably, the percentage intensity of nuclei containing low levels of adducts remained similar in all the subgroups i.e. animals given B(a)P [BP(+24h), BP(+48h), BP(+96h), BP(+144h)] and B(a)P + curcumin [BP(+48h) + C 24 h, BP(+96h) + C 72 h, BP(+144h) + C 120 h]-treated subgroups (Figs. 2A and 2B). Together, results suggest that dietary curcumin led to enhancement of decrease in nuclei containing high and medium levels of adducts in the liver whereas in the lungs a curcumin-mediated enhanced decrease was mainly observed in nuclei containing high levels of adduct(s).

Such systems have been used for many years in the assessment of the effects of novel pharmacological agents on cardiovascular electrophysiological and contractile function ( Habeler et al., 2009). An advantage of these integrated models is that they can maintain their physiological integrity for long periods of time. Such models have yet to be used in research in atherosclerotic cardiovascular disease, but they may lend themselves well to modelling the long-term disease processes which occur in smoking-related

Cell Cycle inhibitor cardiovascular disease. Clearly, a number of in vitro cardiovascular disease models have the potential for use in an approach to assess the biological effects of cigarette smoke from modified cigarettes, and these have been summarised in Table 2. What we have not discussed in this article are the practicalities of use of these models, particularly in terms of

model validation and experimental standards. With respect to the latter, any data and conclusions derived from the use of these models would have greater strength if studies were conducted following the principles of Good Laboratory Practise, which would ensure the quality, integrity and reproducibility of experimental findings ( Gupta et al., 2006). Model validation is an area which needs a great deal of development in order to ensure that the C59 wnt concentration models used Telomerase are fit-for-purpose, in terms of the model used being relevant to the disease being examined and linked to pathogenic processes. Validation would further ensure that data from models was robust, reproducible

and repeatable and that similar findings could be obtained from independent laboratories using the same model and test agents. Of further importance is verification of the identity of the cells used in any given model, to ensure that they are in fact authentic and what the investigator believes them to be ( Freshney, 2008). While in vitro models are powerful assessment tools, a thorough testing strategy may be enhanced with in vivo models. In the realm of cardiovascular disease studies, many animal models have been used over several decades and include a range of species from pigeons to non-human primates. Early animal models relied on atherogenic diets to drive the pathogenesis of cardiovascular disease and typically were time-consuming and expensive. However, important understandings of disease processes resulted from the use of these models. Current in vitro models are poor predictors of events that lead to plaque formation, destabilisation, rupture and thrombotic events.

This causes a drop in the total intratracheal pressure (Buck and Keister, 1955, Buck and Keister, 1958, Buck and Friedman, 1958 and Hetz et al., 1994). In the following flutter phase single spiracles open and close rapidly. Gas exchange works here due to convection and CHIR-99021 chemical structure diffusion. Small amounts of O2 are inhaled to sustain a certain low level of PO2 for a minimum O2 delivery to the insect’s metabolizing tissues (Hetz and Bradley, 2005 and Lighton, 1996). The CO2 level keeps rising in the hemolymph during the flutter phase, as only small amounts of CO2 are exhaled (Buck,

1958). As accumulated CO2 reaches a trigger threshold, a massive amount exits from the tracheal system to the environment in the open-spiracle phase (Lighton, 1996 and Schneiderman and Williams,

1955). CO2 is assumed to act directly at the spiracular muscles, with little central nervous control (Hoyle, 1961); however, Bustami and Hustert, 2000, Bustami et al., 2002 and Woodman et al., 2008 found contrary evidence. Discontinuous Maraviroc clinical trial gas exchange was hypothesized to be an adaptation aimed at minimizing water loss from the tracheae (hygric model, Chown, 2002, Chown et al., 2006a, Dingha et al., 2005, Duncan et al., 2002b, Hadley, 1994, Kivimägi et al., 2011, Williams and Bradley, 1998, Williams et al., 1998 and Williams et al., 2010), though findings by Contreras and Bradley, 2009, Gibbs and Johnson, 2004 and Sláma et al., 2007 call into question the universal validity of this model. Other explanations suggest that it developed to allow sufficient gas exchange in subterranean, CO2 rich environments (chthonic model, Lighton and Berrigan, 1995). A combination of these two models is the hygric-chthonic hypothesis (Lighton, 1998). An alternative explanation suggests that it minimizes old oxygen toxicity (Bradley, 2000 and Hetz and Bradley, 2005). The variation of respiration patterns has been well investigated

in different species (Basson and Terblanche, 2011, Chown et al., 2006a, Groenewald et al., 2012, Klok and Chown, 2005, Kovac et al., 2007, Nespolo et al., 2007, Terblanche et al., 2008a and Williams et al., 2010). Such an analysis is lacking in vespine wasps. This is especially interesting because Vespula sp. show an overall higher level and a steeper incline in resting metabolism with increasing ambient temperature (high Q10) than many other insects (see Käfer et al., 2012). In this paper, therefore, we investigated the characteristics of the respiration patterns of vespine wasps, Vespula sp., over their entire viable temperature range. We compare the specific features of their gas exchange patterns with other flying and nonflying insects. Respiration of adult insects is accomplished by a combination of passive diffusive gas exchange and active convective ventilation (Jõgar et al., 2011, Lighton, 1996 and Terblanche et al., 2008b). Ventilatory movements are usually observed via automated optical activity detection.

1–2.3 μM on HL-60 cells. Regarding to normal cells (PBMC), IC50 values ranged from 3.2 to 13.4 μM and were less pronounced than those found in cancer cells. As shown in Fig. 2A, compounds 2, 3 and 4 caused reduction in HL-60 cell number in the concentration of 2 μM after 24 h treatment and evaluation by trypan blue exclusion test (46.7 ± 2.0, 43.0 ± 2.1 and 48.5 ± 3.3 × 104 cells/mL, respectively) when compared to control cells (65 ± 5.5 × 104 cells/mL) (p < 0.05), while no differences between the compounds were noticed (p > 0.05). The positive control Dox also caused a significant reduction on viable cell population

(42.2 ± 1.0 × 104 cells/mL, p < 0.05). Interestingly, though all compounds has decreased cell number after 24 h exposure,

none of them altered viability of the check details remaining cells, since it was not noticed statistically significant differences in viable and non-viable cells in comparison to control ( Fig. 2B). The cytotoxicity is not related to the membrane lysis of leukemia cells, since compounds 3 and 4 did not led to membrane disruption or increased DAPT mouse fluorescence after ethidium bromide incorporation. The exception was the compound 2 (2 μM), which induced a slight but significant decreasing in cells with intact membranes (93.0 ± 1.6%, p < 0.05) ( Fig. 2C). Since sesquiterpene lactones are known inhibitors of enzymes and cellular processes, we investigated whether the inhibition of cell proliferation is related to DNA synthesis inhibition using the BrdU assay. This method revealed that

all compounds were able to reduce the BrdU incorporation, presenting the compound 4 the highest potential to diminish BrdU-positive cells in both dose tested (1 μM and 2 μM, 28.5 ± 2.2% and 28 ± 1.9%, respectively) PD-1 antibody inhibitor in comparison to negative control (51.4 ± 3.15%). To define the mechanism responsible for the action of santonin derivatives involved on HL-60 cell death, cell-cycle distribution was assessed after 24 h and 48 h of treatment (Fig. 3A and B). A significant inhibition on HL-60 cell-cycle progression was observed within 24 h, where Dox (37 ± 3.4%), compound 3 (7.6 ± 0.5% and 9.0 ± 0.9%) and 4 (9.0 ± 0.9% and 8.6 ± 9.6%) (1 and 2 μM, respectively) caused an increasing of cells in G2/M phase when compared to untreated cells (3.4 ± 0.5%). On the other hand, 48 h exposure provoked G2/M reduction [(2.6 ± 0.7% and 1.5 ± 1.0%), (1.7 ± 0.3% and 1.5 ±.0.5%) and (0.6 ± 0.2% and 1.0 ± 0.8%), for compounds 2, 3 and 4, respectively] when compared to negative control (5 ± 0.8%) (Fig. 3C, p < 0.05), findings indicating time and concentration dependent activity of the molecules. Interestingly, only compound 2 at highest concentration was able to increase sub-G0/G1 DNA content after 24 h (34 ± 4.8%, indicated in pink part) in comparison with control (13 ± 1.3%) ( Fig. 3D). However, after 48 h exposure, α-santonin derivatives 3 and 4 also caused increasing on DNA fragmentation [(45.3 ± 1.2% and 91.0 ± 2.0%) and (64.4 ± 1.