The results showed that all three specific see more growth rates tested yielded approximately the same maximum

ODs (40–50), which were also similar to those obtained with the constant feeds. In these experiments, glycerol concentrations were generally high from the start of the feeding, due to higher feeding speeds, indicating that cells are not able to exhaust all the glycerol added to the culture medium. Taking into account the results of both feeding profiles, the selected feeding profile for hSCOMT induction fermentation was a constant feed of 1 g glycerol/L/h. As mentioned above, for the final fermentations a constant feed of 1 g glycerol/L/h was used, with a higher (50 g/L) initial concentration of tryptone in order to compensate the possible tryptone limitation during the fed-batch phase. All other bioprocess parameters remained unaltered. Firstly, a fermentation without induction was performed, in order to determine the starting point of the stationary phase with this new medium formulation, and consequently the start of the feeding (Fig. 5). As seen in Fig. 5, the stationary phase was reached at about 8 h into the fermentation, and that was the time chosen to initiate the feeding. However, and since there was no significant increase in cell growth after this point, we decided to initiate the feeding 1 h earlier (at 7 h) in the subsequent experiment,

with IPTG induction. The induction was carried out 1 h after starting of the feeding, for 4 h. In this fermentation, glycerol quantitation assays were carried out as mentioned

above http://www.selleckchem.com/products/s-gsk1349572.html and as expected, glycerol consumption profile was very similar to the previous assays carried out with this feeding profile, however in this case, glycerol concentration was low just from the beginning, and after 2 h of feeding, the concentrations remained the same in both replicates (data not shown). Cytometry assays were carried out as explained above, and the results for these fermentations can be seen in Fig. 6 (only for the first replicate). As the results Montelukast Sodium show, the percentage of viable cells at the end of the fermentation are relatively high, between 84% and almost 90%. For the enzymatic assay, samples were taken every 2 h after induction (until 6 h of induction), and treated according to the method described in Section 2.2.3. Specific activity results are plotted in Fig. 5, and as we can observe an increment in activity is achieved during 6 h after induction from 56 nmol/h/mg to 442.34 nmol/h/mg. In recent years, several attempts have been performed to obtain a large quantity of active and pure hSCOMT. One of the most effective ways of enhancing recombinant protein production is the application of a fed-batch process, which highly increases cell density and, subsequently, protein production. In this work, a fed-batch bioprocess was developed for hSCOMT biosynthesis.

An endoscopic response is a decrease from baseline CDEIS score of at least 4 or 5 points. The CDEIS has been used in trials of corticosteroids, thiopurines, and TNF antagonists. In the MUSIC (Endoscopic Mucosal Improvement in Patients With Active Crohn’s Disease Treated With Certolizumab Pegol) study of certolizumab pegol in Crohn’s disease,

maintenance of improvement between weeks 10 and 54, based on individual patient data, was found in 70% of those who responded (decline in CDEIS >5) and those with complete remission (CDEIS<3), and in more than 40% of those with remission (CDEIS<6).43 The SES-CD (Table 6) correlates VE-821 clinical trial well with the CDEIS, with a correlation coefficient r = 0.920

and excellent interobserver reliability (k coefficients 0.791–1.000). This score was developed to meet the need for a reliable, easy-to-use endoscopic scoring instrument for Crohn’s disease, one that by contrast would be less complex than the CDEIS. Selected endoscopic parameters (ulcer size, ulcerated and affected surfaces, stenosis) were scored from 0 to 3, whereby SES-CD = 0 equates to absence of ulcers. 41 No cutoff values have been determined for the SES-CD, and there is no definition of mucosal healing. The Rutgeerts Postoperative Endoscopic Index (Table 7) determines the severity of endoscopic disease recurrence at the anastomosis and in the neoterminal ileum after ileocolic resection.42 and 44 selleck chemicals llc The severity of endoscopic recurrence predicts clinical recurrence, so it has gained popularity.42 In the year after ileocolic resection, patients with a Rutgeerts score of 0 or 1 have a low risk of clinical recurrence (20% at 3 years follow-up) compared with Sinomenine those patients who have a score of grade 3 or 4 (92% at 3 years follow-up). Level 2 is associated with an intermediate risk of clinical recurrence, but the definition of grade 2 is more subjective and is exposed to variability. This index has also been incorporated into

a randomized clinical trial. In the Post Operative Crohn’s Endoscopic Recurrence study, it was shown that treating according to the risk of recurrence with a 6-month postoperative colonoscopy and treatment step up for those who had a Rutgeerts score ≥i2, is significantly superior to drug therapy alone in preventing postoperative recurrence.45 The colonoscopic assessment of mucosal healing has proved increasingly important in the management of both UC and Crohn’s disease. All clinicians should strive for this goal. There is evidence for a decrease in corticosteroid use, decreased hospitalization, an increase in sustained remission, and a decrease in the need for surgery. Further advancements with surrogate noninvasive markers for mucosal healing may help to overcome existing limitations and need for colonoscopy.

Methanol, ethanol, 1-propanol, 2-propanol, dipotassium hydrogen phosphate (K2HPO4), potassium dihydrogen phosphate (KH2PO4) and potassium phosphate (K3PO4) were purchased at Vetec (Rio de Janeiro, Brazil). The alcohols have purities higher than 99 wt.%. Selleck Buparlisib The phosphate salts present had purity levels

higher than 98 wt.%. The l-ascorbic acid (>98 wt.%) was acquired at Labsynth (São Paulo, Brazil) and vanillin (>99 wt.%) was purchased at Sigma–Aldrich. Ultrapure water, double distilled, passed by a reverse osmosis system and further treated with a Milli-Q plus 185 water purification apparatus, was used. The vanilla diet pudding Dr. Oetker was purchased at a regular supermarket in Aracaju, Brazil (http://www.oetker.com.br/?actA = 2111&produtoID = 138). KRX-0401 supplier The ATPS were formed using aqueous solutions of alcohols (methanol, ethanol, 1-propanol and 2-propanol) at 80 wt.% and distinct aqueous solutions of inorganic potassium phosphate salts (K3PO4, K2HPO4 and the phosphate buffer solution KH2PO4/K2HPO4 – Henderson–Hasselbalch equation equivalents = 1.087) at ca. 40 wt.%.

The phase diagrams were determined through two different experimental methodologies well described in literature, the cloud point titration method ( Merchuck et al., 1998, Neves et al., 2009, Ventura et al., 2009, Ventura et al., 2011 and Ventura et al., in press) and the turbidometric titration method ( Aqueous Two-Phase Systems, 2000 and Freire et al., 2012) at (298 ± 1) K

and atmospheric pressure. The tie-lines (TLs) were obtained using a gravimetric method originally applied by Merchuck and co-workers (Merchuck et al., 1998) and already validated in previous studies (Neves Non-specific serine/threonine protein kinase et al., 2009 and Ventura et al., 2009). Each tie-line (TL) was determined by the application of the lever-arm rule. For that purpose, the experimental solubility curves were correlated using the following Eq. (1), equation(1) Y=Aexp[(BX0.5)-(CX3)]where Y and X, are the alcohol and salt mass fraction percentages, respectively, and A, B and C are the regression constants. The partitioning systems for l-ascorbic acid were prepared using graduated centrifuge tubes by weighing the appropriate amounts of alcohol (at ca. 50 wt.%), inorganic salt (at ca. 15 wt.%) and l-ascorbic acid (2.8 mg). To prepare the vanillin partitioning systems, vials with the same weight fractions of alcohol and inorganic salt were prepared. An aqueous solution of vanillin (concentration of ca. 1.0 g.dm−3) was used as the aqueous phase. Afterwards, the mixtures were gently stirred and centrifuged at 3,000 × g for 10 min. The extraction systems were placed at (298 ± 1) K, for at least 18 h, to reach the equilibrium and the consequent and complete partitioning of the antioxidants. The vials were closed during this period to avoid the alcohol vaporisation.

It is difficult to determine the individual arsenic species in order of their toxicity, because the toxicity of these chemical forms is very different not only in different organisms but even between organs. One factor that makes arsenic more interesting is that arsenic is an essential Selleckchem EGFR inhibitor element for some animals, like rats and goats (Püssa, 2008 and Ratnaike, 2003) and interindividual susceptibility in humans to the adverse effects caused by arsenic compounds has been reported (Huang et al., 2004). The initiation and progression mechanisms of human carcinogenesis caused by arsenic

exposure are still not entirely clear (Shi, Shi, & Liu, 2004). However, chronic exposure to inorganic arsenic not only causes, but also can evoke hypertension, skin lesions, diabetes and cardiovascular disease and furthermore it can affect the vascular system (Hughes, 2002 and Jomova et al., 2011). Acute exposure to high levels of arsenic can cause cardiomyopathy, hypotension, gastrointestinal discomfort, vomiting, diarrhea, bloody urine, anuria, shock, convulsions, coma and in death

in the most severe cases (Hughes, 2002 and Jomova et al., 2011). According to the International Agency for Research Selleckchem CHIR 99021 on Cancer (IARC) arsenic is a class I carcinogen (International Agency for Research on Cancer, 1987). In 2004 IARC declared that arsenic could cause lung, skin and urinary bladder cancer in humans (International Agency for Research on Cancer, 2004). In 2010, the Joint FAO/WHO Expert Committee on Food Additives (JECFA) estimated that BMDL0.5 for inorganic arsenic species would be 3 μg/kg bw/day (Joint FAO/WHO Expert Committee on Food Phosphatidylinositol diacylglycerol-lyase Additives, 2010). This conclusion replaced the old PTWI-value for inorganic arsenic (15 μg/kg bw/week) which had been established in 1989. The European Food Safety Authority (EFSA) set the BMDL0.1 value at 0.3 – 8 μg/kg bw/day in 2010

(European Food Safety Authority, 2010). At present, there are no regulations about organic or inorganic arsenic species in food or beverages except for that in drinking water. In 1993, WHO provided a reference value of 10 μg/L of total arsenic compounds in drinking water, previously the reference value had been set at 50 μg/L (World Health Organization, 1993). In 2008 the Data Collection and Exposure Unit (DATEX) of EFSA collected information on the arsenic levels in food from the EU member states and Norway (EFSA, 2010). According to the DATEX survey, the total arsenic level was highest in fish and seafood and miscellaneous dietary products. The miscellaneous group consisted of diverse foodstuffs, e.g. algae, algae based food supplements, spices, herbs, different baby foods and formulas. It is well-known that a significant part of total arsenic in fish and seafood exists in the organic arsenic forms, particularly arsenobetaine (Nam et al., 2010, Sloth et al., 2005 and Suner et al.

, 2007). If error cannot

be avoided (e.g., if all available samples were obtained post-fast), it is important to assess accuracy of exposure characterization by calculating sensitivities and specificities (Jurek et al., 2006). Sensitivity GDC-0199 concentration is the probability of correctly classifying an individual as having high level of exposure, if that person truly belongs in the high exposure category. Specificity is the probability of correctly assigning low exposure to a participant who truly has a low level of exposure. Estimates of sensitivity and specificity may be calculated for a single urine sample, using multiple samples per subject as gold standard, since the true sensitivity and specificity for many measures www.selleckchem.com/products/iox1.html is unknown. This can be achieved by randomly selecting a single sample from among each individual’s repeated samples collected over the study (as demonstrated for phthalates in Adibi et al., 2008). In a recent systematic review of the epidemiology literature on phthalates and associations with obesity, diabetes, and cardiovascular disease, Goodman et al. (2014) found that of 26 available studies, all but three relied on a single

measure of phthalates. Similarly, in a systematic review of BPA and obesity, diabetes, and cardiovascular disease, LaKind et al. (2014) found that of 45 available studies, all but four relied on a single measure of BPA. Yet the intra-individual variability for BPA is large (with ICCs ranging from 0.10 to 0.35) (Lassen et al., 2013 and Teitelbaum et al., 2008), and multiple measures of exposure are needed to describe a person’s long-term exposure. The ICCs for phthalates have been reported to be higher than for BPA (e.g., ICC values range

from 0.18 to 0.61 for mono-ethyl phthalate, from 0.21 to 0.51 for mono-isobutyl phthalate, and from 0.08 to 0.27 for mono-(2-ethylhexyl) phthalate [reviewed in Goodman et al., 2014]), but intra-person variability Rebamipide is still large. Recently, Attfield et al. (2014), in a study of variability of urinary pesticide measures in children, observed that a study with only a small number of samples from each study participant “…may lead to a high probability of exposure misclassification by incorrect quantile assignment and offer little assurance for correctly classifying the exposure into a specific category. The above considerations permit dividing the available body of literature into the following tiers (Table 1). Tier 1 includes studies in which exposure assessment is based on sufficient number of samples per individual to estimate exposure over the appropriate duration, or through the use of adequate long-term sampling (e.g., multiple 24-hour urine collections). To be included in Tier 1, studies should assess error by calculating measures of accuracy (e.g., sensitivity and specificity) and reliability (e.g., ICC). It is possible that for some chemicals, one sample may be sufficient to fully characterize exposure.

When describing higher-codability events, speakers showed only a small preference

for the agent over the patient, and properties of the agent were weak predictors of the magnitude of this preference. In lower-codability events, on the other hand, the pattern of early fixations was primarily determined by Agent codability: speakers shifted their attention very rapidly to “easy” agents and away from “hard” Selleckchem Anti-cancer Compound Library agents. As in Experiment 1, this result suggests that speakers attempted to select a starting point based on character accessibility when they could not easily select a starting point based on their construal of the gist of the event. It also extends Kuchinsky and Bock’s (2010) observations about the influence of relational factors on selection of starting points to the timecourse of sentence formulation. The benefits of early encoding of event gist carried over to later time windows as well. In

higher-codability events, speakers directed their attention to the agent relatively quickly after 400 ms. By comparison, the strong preference to fixate the agent in lower-codability events before 400 ms resulted in a less consistent pattern of fixations: rapid shifts of attention to the agent within 400 ms of picture onset were followed by an extended time window Selleckchem ALK inhibitor where speakers fixated the patient (as in Experiment 1, large shifts of attention from one character to another suggest that the two characters were encoded sequentially). As a result, agent-directed fixations after 400 ms also showed a joint influence of Event and Agent codability: speakers were able to deploy their attention to the agent and finally shift their gaze to the patient earlier in “easier” events than in “harder” TCL events (this effect was stronger than in Experiment 1, which showed a main effect of Event codability but no interaction of Event codability with Time bin). Critically, the effect of

structural primes on formulation was different from the effect of lexical primes in Experiment 1: the structural primes produced shifts in planning patterns that resembled the effect of Event codability on formulation and thus were consistent with hierarchical incrementality. As predicted, active primes reduced the proportion of agent-directed fixations within 400 ms of picture onset in active sentences, suggesting a very early effect of structural processes on visual inspection of an event. The interaction with Event codability in this time window indicates stronger facilitation of early relational encoding when both conceptual and linguistic structures were easy to generate. After active primes, speakers also quickly directed their gaze to the agent after 400 ms and to the patient before speech onset.

BD. (LIFE 09 ENV/IT/000078). We thank Mihej Urbančič for assistance with fieldwork. We are also grateful to two anonymous reviewers

for constructive criticism and helpful comments on the manuscript. “
“Stemwood production is influenced find more by climate, nutrients, and water, but is also determined by the amount of light intercepted and the photosynthetic efficiency of canopies (Vose and Allen, 1988). Canopy structure throughout the vertical and horizontal profiles can be described by biophysical forest parameters such as leaf area and tree height. Leaf area index (LAI) is defined as the total one-sided area of leaf tissue per ground surface area (Watson, 1947). It plays an important role in several key ecosystem processes by the exchange of energy and gases (e.g., CO2 and water-vapor fluxes) between terrestrial ecosystems and the atmosphere.

It is also central to describing rainfall interception. As a result, leaf area varies along with hydrological, biogeochemical, and biophysical processes, either due to natural stand development or forest management practices (e.g., thinning, RG7420 research buy fertilization, and vegetation control). Along with leaf biomass, leaf area has a strong relationship with productivity (Cannell, 1989). In loblolly pine (Pinus taeda L.), for example, leaf biomass dynamics are dependent on phenology, climatic conditions, site factors and stand density, thus LAI represents a measure of site occupancy that integrates tree size, stand density and site resource supply ( Vose and Allen, 1988). Based on these relationships, forest managers have observed crown development and

leaf production as responses to fertilization and thinning; such responses are consequently related to carbon accumulation and tree growth ( Albaugh et al., 1998, Carlyle, 1998 and Martin and Jokela, 2004). Traditional approaches to directly estimate leaf area index, such as using destructive sampling, although very accurate, are labor intensive, time consuming, and costly. The resulting paucity of samples limits their utility for forest management. The use of remote sensing technologies to monitor, and therefore to improve the management of forest resources Phloretin at regional and global scales has increased exponentially over the last 30 years (Lefsky et al., 2002b, Lu, 2006 and Lutz et al., 2008). Previous research has shown that satellite data can be used to estimate LAI accurately in areas where LAI has been empirically related to satellite-measured reflectance values (Curran et al., 1992, Gholz et al., 1997, Jensen and Binford, 2004 and Flores et al., 2006). Green vegetation amounts and leaf area index have been associated with spectral reflectance, and frequently with vegetation indices. Nonetheless, researchers have observed that optically-derived vegetation indices reach an asymptote or saturation point when LAI values are on the order of 3–5 (Spanner et al., 1990b, Turner et al.

, 2004) to >1 mg/ml (Kimura et al., 2000). Modification of sulfated oligosaccharides with a relatively short alkyl chain (dodecyl) was employed in glycoside 3 (Table 1) which exhibited a favorable IC50 value and no cytotoxicity Nutlin-3 datasheet (Table 2), however, due to modest virucidal activity this compound was not extensively studied. More pronounced virucidal activity was observed in PG545 and it is difficult to compare it with NMSO3 since no data on the virucidal activity of this compound was reported. We found that the virucidal activity of PG545 was decreased in the presence of FCS in culture medium, and this observation is not surprising since

sterols can interact with several serum proteins including apolipoproteins. More importantly, because PG545

would need to target RSV infecting cells of the airway, we tested whether the antiviral activity Selleckchem Selisistat of this compound is modulated in the presence of human nasal secretions. We found that pooled preparations of nasal secretions can inhibit RSV infectivity. The anti-RSV activity of nasal secretions could be exerted by some components of this body fluid such as surfactant proteins (Ghildyal et al., 1999), antimicrobial peptides (Laube et al., 2006 and Kota et al., 2008), mucins (Rubin, 2002), or cholesteryl esters (Do et al., 2008). Moreover, we found that human nasal secretions reduced anti-RSV activity of PG545, however, this inhibitory effect could be overcome by using higher concentrations of PG545. Further studies employing a model of RSV infection of well-differentiated cultures of human airway epithelium (Zhang et al., 2002) are needed to assess modulation of anti-RSV activity of PG545 by airway mucus. The capability of PG545 and related glycosides to interact with serum proteins did not seem to limit their in vivo application. In fact, the presence of the lipophilic moiety in PG545 and related glycosides helped to overcome two major disadvantages associated with in vivo usage of sulfated oligo- and polysaccharides,

i.e., it Clomifene greatly attenuated their anticoagulant activity and prolonged the half life of these compounds in the body (Johnstone et al., 2010 and Dredge et al., 2011). Due to the presence of sulfate groups in PG545 and related glycosides, these compounds can inhibit the interaction between a plethora of different proteins and sulfated GAGs. Thus, interference of PG545 with the activity of vascular endothelial and fibroblast growth factors inhibited angiogenesis, a key process in tumor development, while binding to heparanase, an enzyme abundantly expressed on neoplastic cells, limited their metastasis (Dredge et al., 2010, Dredge et al., 2011 and Johnstone et al., 2010). Both these functional features confer potent anti-cancer activities on PG545 (Dredge et al., 2011).

To screen for the best-performing siRNA of each group, we employed a dual-luciferase assay-based system. The respective target sequences were individually inserted into the 3′ UTR of a plasmid-located Renilla luciferase

gene. The DNA polymerase, pTP, IVa2, hexon, and protease siRNAs, together with Lonafarnib mouse the respective reporter vectors, were used to co-transfect HEK293 cells. Knockdown of Renilla luciferase expression in relation to the expression of a firefly luciferase gene located on the same plasmid was determined in dual-luciferase assays. The silencing capacity of the E1A siRNAs was assessed in A549 cells because promoter activities of the reporter vectors turned out to be altered upon silencing of the endogenous E1A gene present in HEK293 cells ( Graham et al., 1977) (data not shown). For all target mRNAs,

we identified siRNAs enabling a knockdown of ⩾78% at a concentration of 30 nM ( Fig. 1). The best-performing siRNAs of each group, i.e., pTP-si8, Pol-si2, Hex-si2, E1A-si3, Iva2-si2, and Prot-si1, were selected for further characterization. The dual-luciferase assay-based screening system was employed to select the best-performing siRNAs of each group. Next, we investigated whether the selected siRNAs were able to knockdown gene expression during an adenovirus infection of A549 cells. Cells were transfected with the siRNAs at a concentration of 10 nM, and then infected with Ad5 at an MOI of 0.01 TCID50/cell. Target mRNA levels were determined check details by RT-qPCR, using primers specific for the individual mRNAs (Fig. 2A).

The highest silencing rates (93–97%) were observed for the E1A-, DNA polymerase-, pTP-, and IVa2-targeting Racecadotril siRNAs. Silencing of the hexon and protease genes was less pronounced (79–87%). Except for the difference in residual hexon and pTP mRNA levels, the differences between hexon or protease mRNA levels and those of all other early genes were statistically significant. As the pTP, DNA polymerase, and IVa2 mRNAs share a common 3′ part (Supplementary Fig. 1), and the DNA polymerase target site is also part of the pTP mRNA, IVa2- and DNA polymerase-directed siRNAs were therefore expected to concomitantly silence pTP/DNA polymerase/IVa2 and pTP/DNA polymerase, respectively. Furthermore, siRNA-mediated silencing of early genes was expected indirectly to affect the expression of those middle or late genes for which expression is known to depend on early viral gene products. Thus, we also determined the effect of the E1A-, pTP-, DNA polymerase-, and IVa2-targeting siRNAs on the expression of the other genes. Silencing of E1A resulted in a marked reduction in the expression of all other genes (Fig. 2B). This can be attributed to the central role of E1A in activating the expression of downstream genes. Silencing of the E1A gene actually resulted in a greater reduction in the expression of hexon and protease than did direct silencing of these genes by the hexon and protease siRNAs (compare Fig. 2A and B).

Akt activation plays a key role in cell proliferation, cell cycle progression, and apoptosis [10]; thus, PI3K/Akt signaling is important for cell survival. Panax ginseng Meyer is one of the most popular herbal medicines in Korea, and has long been used in Asian countries for stimulating immunity and inhibiting

various cancers [11], [12] and [13]. Ginsenosides are active compounds present in ginseng that are known to have antioxidative, anti-inflammatory, and anticancer activities [14]. Ginsenoside Rb1, a known phytoestrogen, shows anti-inflammatory activity in smooth muscle cells [15] and inhibits interleukin-1β-induced apoptosis in human chondrocytes [16]. Ginsenoside Rg3 exerts neuroprotective, anti-inflammatory, and antioxidative effects [17] and [18]. Although the role of ginseng in regulating the development of cancer is well defined, the mechanism by which it GS-1101 in vitro protects brain cells from oxidative stress is not well understood. Recent studies have revealed that ginseng upregulates ER-β expression in vitro and in vivo [17] and [19]. Previously, we reported that Korean Red Ginseng (KRG)-induced ER-β expression inhibits oxidative stress-induced apoptosis

in mouse brain and SK-N-SH neuroblastoma cells by inhibiting PADI4 expression [17]. However, the downstream signaling effector molecules of ER-β have not been explored. Thus, the aims of this study were to identify signaling effector molecules immediately downstream of ERβ and to understand how KRG-induced ER-β expression regulates Amoxicillin apoptosis via PI3K/Akt signaling www.selleckchem.com/products/r428.html in oxidative stressed brain cells. Human neuroblastoma SK-N-SH cells (catalog number HTB-11; ATCC, Manassas, VI, USA) were cultured in RPMI 1640 (Lonza, Walkersville, MD, USA) media containing 10% FBS, 1% penicillin-streptomycin (10,000 U penicillin/mL, 10,000 μg streptomycin/mL), 1mM HEPES, 1mM sodium pyruvate, 4.5 g/L glucose, 1.5 g/L bicarbonate, and 2mM L-glutamine at 37°C, and 5% CO2. KRG extract was manufactured by Korea Ginseng Corporation (Seoul,

Korea) by steaming and drying 6-year-old roots from Panax ginseng Meyer and analyzed as described previously [17]. The ginsenoside content of KRG extracts used in this study was: Rg1 0.71 mg/g, Re 0.93 mg/g, Rf 1.21 mg/g, Rh1 0.78 mg/g, Rg2(s) 1.92 mg/g, Rg2(r) 1.29 mg/g, Rb1 4.62 mg/g, Rc 2.41 mg/g, Rb2 1.83 mg/g, Rd 0.89 mg/g, Rg3(s) 2.14 mg/g, and Rg3(r) 0.91 mg/g. Specific inhibitors of ER-β (PHTPP: catalog number sc-204191) and Akt (inhibitor VIII; catalog number sc-2002048) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). The PI3K-specific inhibitor LY294002 (catalog number L9908) was purchased from Sigma–Aldrich (St Louis, MO, USA). SK-N-SH cells were treated with KRG extract for 48 h and subsequently treated with 5μM PHTPP [20], 80μM LY294002 [21], or 50μM Akt inhibitor VIII for 5 h.