ß-cyclodextrin functionalized meso-/macroporous magnetic titanium dioxide adsorbent as extraction material combined with gas chromatography-mass spectrometry for the detection of chlorobenzenes in soil samples.

A high-performance and selective adsorbent was developed for simultaneous extraction of 6 chlorobenzenes residues in soil samples by using magnetic solid phase extraction (MSPE) combined with automated SPE followed by gas chromatography-mass spectrometry (GC-MS). The adsorbent was synthesized by grafting carboxymethyl-ß-cyclodextrin (CM-ß-CD) on the surface of porous core-shell magnetic Fe3O4@flower like TiO2 microspheres (Fe3O4@fTiO2-CMCD), used as a carrier. The main factors (adsorbent amount, adsorption time, elution solvent, elution volume, and elution flow rate) affecting the extraction efficiency were investigated in detail. The adsorbent exhibited high loading capacity (25.6 mg g(-1) for 1,3-dichlorobenzene). This maybe due to meso-/macroporous TiO2 having high specific surface area; as a carrier of the ß-cyclodextrin film, it could obviously increase the number of recognition sites. The newly developed adsorbent also showed good selectivity towards chlorobenzenes based on host-guest interactions between ß-cyclodextrin (on adsorbent's surface) and targets, which can minimize complex matrix interference in soil samples. The proposed method was successfully applied for the analysis of environmental soil samples with recovery ranging from 87.3 to 104.3%. All target compounds showed good linearities with correlation coefficients (r) higher than 0.996. The limits of quantitation for the 6 CBs were 0.03-0.09 µg kg(-1). These findings confirmed meso-/macroporous structure Fe3O4@fTiO2-CMCD as a highly effective extraction material for use in trace CB analyses in complex soil samples.

Separation and enrichment of six indicator polychlorinated biphenyls from real waters using a novel magnetic multiwalled carbon nanotube composite absorbent.

A novel and effective magnetic multiwalled carbon nanotube composite for the separation and enrichment of polychlorinated biphenyls was developed. Fe3 O4 @SiO2 core-shell structured nanoparticles were first synthesized, then the poly(sodium 4-styrenesulfonate) was laid on its surface to prepare the polyanionic magnetic nanoparticles. The above materials were then grafted with polycationic multiwalled carbon nanotubes, which were modified by polydiallyl dimethyl ammonium chloride through the layer-by-layer self-assembly approach. Its performance was tested by magnetic solid-phase extraction and gas chromatography with mass spectrometry for the determination of six kinds of indicator polychlorinated biphenyls in water samples. Under optimal conditions, the spiked recoveries of several real samples for six kinds of polychlorinated biphenyls (PCB28, PCB52, PCB101, PCB138, PCB153, PCB180) were in the range of 73.4-99.5% with relative standard deviations varying from 1.5 to 8.4%. All target compounds showed good linearities in the tested range with correlation coefficients higher than 0.9993. The limits of quantification for six kinds of indicator polychlorinated biphenyls were between 0.018 and 0.039 ng/mL. The proposed method was successfully applied to analyze polychlorinated biphenyls in real water samples. Satisfactory results were obtained using the effective magnetic absorbent.

Employment of a novel magnetically multifunctional purifying material for determination of toxic highly chlorinated polychlorinated biphenyls at trace levels in soil samples.

In this study, we developed a magnetically multifunctional purifying material for efficient removal of matrix interferences, especially certain organochlorine pesticide (DDT, DDE, and DDD), during the determination of toxic highly chlorinated polychlorinated biphenyls (PCBs) at trace levels in soil samples. The multifunctional adsorbent (CMCD-NH2-MNPs) was prepared by grafting carboxymethyl-ß-cyclodextrin on the surface of amino-functionalized magnetite (Fe3O4) nanoparticles. CMCD-NH2-MNPs has stronger host-guest complexation with DDT, DDE, and DDD, but the same adsorbent shows weaker adsorption ability toward highly chlorinated PCBs (from tetra- to octa-chlorinated PCBs) owing to their steric hindrance effect. Based on this principle, a simple and rapid gas chromatography-mass spectrometry (GC-MS) method was developed for six indicator PCBs (PCB28, PCB52, PCB101, PCB138, PCB153, and PCB180) in soil. Comparative studies were conducted to determine the clean-up efficiency of the following three techniques: (i) Oasis-HLB, (ii) multi-layer silica column, and (iii) dSPE employing CMCD-NH2-MNPs. The results indicate that CMCD-NH2-MNPs as the purification material can easily and effectively remove DDT, DDE, and DDD in soil samples within a short duration of time. The recoveries for highly chlorinated PCBs were in the range of 85.4-102.2%, with RSDs varying between 1.0 and 6.5%. The proposed method was verified as one of the most effective clean-up procedures for the analysis of highly chlorinated PCBs in real soil samples.

Prodigiosin activates endoplasmic reticulum stress cell death pathway in human breast carcinoma cell lines.

Prodigiosin is a bacterial tripyrrole pigment with potent cytotoxicity against diverse human cancer cell lines. Endoplasmic reticulum (ER) stress is initiated by accumulation of unfolded or misfolded proteins in the ER lumen and may induce cell death when irremediable. In this study, the role of ER stress in prodigiosin-induced cytotoxicity was elucidated for the first time. Comparable to the ER stress inducer thapsigargin, prodigiosin up-regulated signature ER stress markers GRP78 and CHOP in addition to activating the IRE1, PERK and ATF6 branches of the unfolded protein response (UPR) in multiple human breast carcinoma cell lines, confirming prodigiosin as an ER stress inducer. Prodigiosin transcriptionally up-regulated CHOP, as evidenced by its promoting effect on the CHOP promoter activity. Of note, knockdown of CHOP effectively lowered prodigiosin's capacity to evoke PARP cleavage, reduce cell viability and suppress colony formation, highlighting an essential role of CHOP in prodigiosin-induced cytotoxic ER stress response. In addition, prodigiosin down-regulated BCL2 in a CHOP-dependent manner. Importantly, restoration of BCL2 expression blocked prodigiosin-induced PARP cleavage and greatly enhanced the survival of prodigiosin-treated cells, suggesting that CHOP-dependent BCL2 suppression mediates prodigiosin-elicited cell death. Moreover, pharmacological inhibition of JNK by SP600125 or dominant-negative blockade of PERK-mediated eIF2α phosphorylation impaired prodigiosin-induced CHOP up-regulation and PARP cleavage. Collectively, these results identified ER stress-mediated cell death as a mode-of-action of prodigiosin's tumoricidal effect. Mechanistically, prodigiosin engages the IRE1-JNK and PERK-eIF2α branches of the UPR signaling to up-regulate CHOP, which in turn mediates BCL2 suppression to induce cell death.

Prodigiosin-induced cytotoxicity involves RAD51 down-regulation through the JNK and p38 MAPK pathways in human breast carcinoma cell lines.

RAD51 is essential for homologous recombination (HR)-mediated repair of DNA double-strand breaks (DSBs) in mammalian cells. RAD51 is an attractive target for anticancer drugs, given high RAD51 levels are frequently observed in many human tumors and associated with increased resistance to DSBs-inducing chemotherapeutics. Prodigiosin is a bacterial tripyrrole pigment with potent anticancer activity and also provokes DSBs. We hereby aimed to elucidate the role of RAD51 in prodigiosin-induced cytotoxicity. Prodigiosin was found to down-regulate RAD51 in multiple human breast carcinoma cell lines irrespective of p53 status. Mechanistically, prodigiosin lowered RAD51 mRNA expression, whereas blockade of proteasome-mediated degradation failed to restore RAD51 levels following prodigiosin treatment. In addition, prodigiosin triggered phosphorylation of JNK and p38 MAPK, while pharmacological inhibition of JNK or p38 MAPK attenuated prodigiosin-mediated inhibition of RAD51 mRNA expression. Lastly, cells with enforced RAD51 expression showed increased resistance to prodigiosin-induced cytotoxicity as well as inhibition of colony formation. Collectively, we conclude that RAD51 down-regulation represents one of the modes of prodigiosin's cytotoxic action, ostensibly by augmenting the genotoxic effect of prodigiosin through suppression of RAD51-mediated HR repair. Our findings further implicate the use of prodigiosin to potentiate the cytotoxicity of DSB-inducing chemotherapeutics through RAD51 down-regulation.

Prodigiosin down-regulates SKP2 to induce p27(KIP1) stabilization and antiproliferation in human lung adenocarcinoma cells.

BACKGROUND AND PURPOSE: High levels of SKP2 are a poor prognostic factor in multiple human cancers and mostly correlate with low p27(KIP1) levels. Prodigiosin is a bacterial tripyrrole pigment with strong pro-apoptotic activity. Induction of cell cycle blockade underlies one of its anticancer actions but the mechanisms involved are unclear. The aim of this study was to explore the role of the SKP2-p27(KIP1) axis in prodigiosin's cytostatic effect on human lung adenocarcinoma cells. EXPERIMENTAL APPROACH: Prodigiosin's effects on cell cycle progression and long-term cell proliferation of human lung adenocarcinoma cells were characterized by flow cytometry and colony formation assay, respectively. Real-time RT-PCR and promoter activity analyses were performed for assessing transcriptional control, while cycloheximide chase analysis evaluated protein stability. Immunoblotting was employed for mechanistic study. KEY RESULTS: Prodigiosin increased p27(KIP1) expression mainly by stabilizing p27(KIP1) through transcriptional repression of SKP2. Importantly, SKP2 overexpression or p27(KIP1) depletion restored the colony forming capacity of prodigiosin-treated cells. Furthermore, prodigiosin induced PKB dephosphorylation, leading to PKB inhibition as revealed by decreased serine 9 phosphorylation of GSK-3ß. Constitutive PKB activation reduced prodigiosin-induced SKP2 repression. Prodigiosin also down-regulated E2F1 (mediates PI3K/PKB-induced SKP2 transcription), but E2F1 overexpression failed to restore SKP2 expression in prodigiosin-treated cells. CONCLUSIONS AND IMPLICATIONS: Transcriptional repression of SKP2 and the consequent accumulation of p27(KIP1) are essential for prodigiosin's antiproliferative action. Mechanistically, prodigiosin induces PKB inhibition to down-regulate SKP2 in a GSK-3ß- and E2F1-independent manner. Our findings further implicate the potential for developing prodigiosin as a novel class of SKP2-targeting anticancer agent.

Anti-inflammatory activity of Flavokawain B from Alpinia pricei Hayata.

Alpinia pricei (Zingiberaceae) is a spicy herb indigenous to Taiwan. A potent anti-inflammatory compound, flavokawain B (FKB), was obtained from A. pricei. FKB significantly inhibited production of NO and PGE(2) in LPS-induced RAW 264.7 cells. Moreover, it also notably decreased the secretion of TNF-alpha. Expression of iNOS and COX-2 proteins was also inhibited by FKB in a dose-dependent manner. FKB blocked the nuclear translocation of NF-kappaB induced by LPS, which was associated with prevention IkappaB degradation, and subsequently decreased NF-kappaB protein levels in the nucleus. Similar anti-inflammatory activities of FKB were observed in an animal assay. NO concentrations in mouse serum rose dramatically from 3.2 to 28.8 microM after mice were challenged with LPS; however, preadministration of 200 mg/kg FKB reduced the NO concentration to 3.8 microM after challenge with LPS. Moreover, FKB strongly suppressed LPS-induced iNOS, COX-2, and NF-kappaB proteins expression in mouse liver.