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A direct competitive fluorescence immunoassay (dc-FIA) and a direct competitive enzyme-linked immunosorbent assay (dc-ELISA) for the screening of dimethyl phthalate (DMP) in water samples were developed. The immunoassays utilise polyclonal antibodies against DMP raised in rabbits. The anti-DMP antibodies were linked to horseradish peroxidase (HRP) and fluorescein isothiocyanate (FITC). Under the optimal experimental conditions, the dc-ELISA has a linear working range of 0.1–2000 ng/ml (R 2=0.993) with a limit of detection of 0.09 ng/ml. In the dc-FIA, the linear working range was 0.05–30 ng/ml (R 2=0.996), and the limit of detection was 0.02 ng/ml, which is approximately four-fold more sensitive than the dc-ELISA using the same antibody and coating antigen. The results show low cross-reactivity with other structurally related compounds. The proposed methods are successfully applied to determine the DMP contaminants with a simple extraction procedure, and good recoveries were obtained. [ABSTRACT FROM AUTHOR]

Abstract: Bendamustine, a hybrid molecule of purine analog and alkylator, induces cell death by activation of apoptosis, DNA damage response, and mitotic catastrophe. Entinostat, a selective class I inhibitor of histone deacetylase (HDAC), exerts anti-tumor activity in various cancer types, including multiple myeloma (MM). We sought to determine the combinatorial effects of bendamustine and entinostat on MM cells. Cell growth assays showed that bendamustine or entinostat inhibited proliferation in a dose-dependent manner, and their combinations synergistically induced growth inhibition in all MM cells tested. An apoptotic-ELISA and western blot assays on PARP cleavage and caspase-8 and caspase-3 revealed that bendamustine in combination with entinostat exhibited a much more potent activity than either agent alone to promote the MM cells undergoing apoptosis in a dose-dependent manner. Flow cytometric analysis found that entinostat exhibited distinct effects on cell cycle progression in different lines and bendamustine mainly arrested the cells at S phase, whereas their combinations dramatically blocked the S cells entering G2/M phase. Furthermore, studies on DNA damage response indicated that phospho-histone H2A.X (P-H2A.X), a hall marker of DNA double strand break, along with phosphorylated CHK2 (P-CHK2) was significantly enhanced by the combinations of bendamustine and entinostat as compared to either agent alone. These molecular changes were correlated with the increases in mitotic catastrophe. Collectively, our data demonstrate that bendamustine in combination with entinostat exhibit potent anti-proliferative/anti-survival activity in MM cells via induction of apoptosis and DNA damage response. Regimens consisting of bendamustine and/or entinostat may represent novel therapeutic strategies against MM. [Copyright &y& Elsevier]

A highly sensitive and specific direct competitive enzyme-linked immunosorbent assay has been developed for the detection of dipropyl phthalate (DPrP) based on antibody-coating format. The specific polyclonal antibodies against DPrP raised in rabbits. The conjugates of the antigen with horseradish peroxidase (HRP) were used as the detectable probe. Under the optimal conditions, the limit of detection of the method was 0.01 ng/mL, and a good quantitative working range of 0.01–16 ng/mL (R2=0.995) was obtained. The cross-reactivities of four structurally related phthalate esters were below 12% and did not interfere significantly the analysis of DPrP. The accuracy of the immunoassay has been evaluated and validated by analyzing real and spiked leachate from plastic food contact materials. The recoveries of DPrP ranged from 85.9% to 109.4%. This developed dc-ELISA was reliable, accurate and highly sensitive. [ABSTRACT FROM PUBLISHER]

Abstract: Trastuzumab (or Herceptin), as the first erbB2-targeted therapy, has been successfully used to treat breast cancer patients with erbB2-overexpressing tumors. However, resistances to trastuzumab frequently occur, and novel strategies/agents are urgently needed to abrogate the resistant phenotype. Our current study explores the potential of SNDX-275, a class I HDAC inhibitor, to overcome trastuzumab resistance and investigates the combinational effects of SNDX-275 and trastuzumab on both sensitive and resistant breast cancer cells. Cell proliferation assays showed that SNDX-275 significantly enhanced trastuzumab-induced growth inhibition in trastuzumab-sensitive, erbB2-overexpressing breast cancer cells. Importantly, SNDX-275 at its therapeutic range re-sensitized trastuzumab-resistant cells to trastuzumab-mediated growth inhibition. SNDX-275 in combination with trastuzumab resulted in a dramatic reduction of erbB3 and its phosphorylation (P-erbB3), and inhibition of Akt signaling. Apoptotic-ELISA and western blot analyses confirmed that the combinations of SNDX-275 and trastuzumab as compared to SNDX-275 alone significantly enhanced DNA fragmentation and induced more PARP cleavage and caspase-3 activation in both trastuzumab-sensitive and -resistant breast cancer cells. Furthermore, co-immunoprecipitation assays revealed that SNDX-275 mainly attenuated the interactions of erbB2 and erbB3 receptors, but had no significant effect on erbB2/IGF-1R or erbB3/IGF-1R associations in the trastuzumab-resistant breast cancer cells. These data indicated that SNDX-275 enhanced trastuzumab efficacy against erbB2-overexpressing breast cancer cells, and exhibited potential to overcome trastuzumab resistance via disrupting erbB2/erbB3 interactions and inactivating PI-3K/Akt signaling. SNDX-275 may be included in erbB2-targeted regimen as a novel strategy to treat breast cancer patients whose tumors overexpress erbB2. [Copyright &y& Elsevier]

Abstract: A direct competitive enzyme-linked immunosorbent assay (ELISA) has been developed for detection of diethyl phthalate (DEP). Protein–hapten conjugate was synthesized to produce polyclonal antibodies against DEP. Experimental parameters were optimized, including immunoreaction conditions, the dilution ratio of horseradish peroxidase (HRP)–antigen conjugate, time of the antibody coated, effect of pH, and ionic strength. The limit of detection was 0.096ng/ml, and the linear range was 0.1–3500ng/ml with a regression coefficient (R 2) of 0.9957. Recoveries were between 96.4 and 106.2%. The cross-reactivities of the anti-DEP antibody to six structurally related phthalate esters were less than 9%. The method was successfully applied to the determination of DEP in tap water, river water (Yangtze River), and leachate from plastic drinking bottles. This immunoassay was highly specific, sensitive, rapid, simple, and suitable for DEP monitoring. The results obtained were compared with those obtained using the high-performance liquid chromatography method. [ABSTRACT FROM AUTHOR]