Anti-proliferative effects of a small molecule inhibitor of CDK AT7519 on chronic myeloid leukemia (CML) cells through halting the transition of cells from G2/M phase of the cell cycle

Pathogenesis of chronic myeloid leukemia (CML) has mostly been studied with regard to the oncogenic role of BCR/ABL fusion; however, recent disclosures have declared that the challenges with the treatment of CML patients would not be resolved until the role of other aberrancies is ignored. Given the involvement of cyclin-dependent kinases (CDKs) in the pathogenesis of CML, the present study aimed to investigate the effects of a multi-CDK inhibitor AT7519 on BCR/ABL-harboring CML-derived K562 cells. Our results showed that AT7519 effectively reduced the survival of K562 and induced its anti-proliferative effect through the induction of G2/M arrest due to elevated p21 and p27. The resulting data also revealed that either direct or indirect suppression of c-Myc using specific c-Myc inhibitor 10058-F4 and selective PI3K inhibitor CAL-101 resulted in a superior cytotoxicity, suggesting that the activation of PI3K pathway could attenuate antileukemic effects of the inhibitor, at least partly, through a c-Mycdependent mechanism. To the best of our knowledge, to date, no study has addressed the effect of autophagy on CML cell response to AT7519, and, herein, we proposed for the first time that the suppression of autophagy boosted AT7519 cytotoxicity against K562. Overall, we suggested that selective CDK inhibitor AT7519 exerted antileukemic effect against CML cells and propose a novel therapeutic application for the inhibitor either as a single agent or in combination with c-Myc and/or PI3K inhibitors.     

MICROWAVE AND CONVENTIONAL BLANCHING EFFECTS ON CHEMICAL, SENSORY, AND COLOR CHARACTERISTICS OF FROZEN BROCCOLI

Broccoli (cv. Empress) obtained from a local supplier was blanched within 15 h of harvest. It was blanched by four methods in covered containers: conventional boiling water (1900 mL, 4 min) (BW), steam (300 mL water, 4 min) (ST), microwave heated in 1 L glass containers (60 ml water, 4 min, 700 W) (MW), and microwave heated in 1 L Seal-a-Meal^TM bags (45 ml water, 4 min) (MWB). Aliquots were frozen at -18C for 4 weeks. Fresh unblanched broccoli peroxidase activity ranged from 389 to 829 units/min; activity was essentially zero immediately after all blanching treatments. The highest reduced ascorbic acid (RAA) content occurred in fresh unblanched broccoli. Some peroxidase regeneration occurred during frozen storage. Immediately after blanching, all blanched broccoli had lower RAA content than control broccoli. MW-blanched broccoli retained the greatest amount of RAA and had appearance, visual color, texture scores, and chroma of florets and stems equivalent to ST-blanched broccoli. MW-blanched broccoli had flavor and general acceptability scores similar to BW-blanched broccoli. After 4 weeks in frozen storage, MW-blanched broccoli had the highest RAA content.     

An evaluation of the Penzym® assay: a rapid method for the detection of β-lactam antibiotics in milk

The Penzym® method, an enzymatic method which allows the detection of β-lactam antibiotics within 20 min, was evaluated using raw whole milk to establish its repeatability and reproducibility for the determination of penicillin G. Three screening thresholds, 0.005, 0.010 and 0.020 iu/ml, were tested by two operators from different laboratories.
Chi-squared(χ²) analysis of the results obtained by each operator for 10 replicate tests of eight penicillin G standards for each screening threshold showed the repeatability of the assay to be satisfactory. Similarly, χ² analysis of the results obtained by each operator for duplicate tests on the same standards assayed on five consecutive days showed that reproducibility for the assay was also satisfactory. It was shown that by altering the volume of milk assayed the sensitivity of the test can be adjusted. The sensitivity of the method to other β-lactam antibiotics used in bovine therapy requires investigation, and a more extensive examination of the repeatability and reproducibility of the assay in several laboratories is needed before it can be recommended for routine use.