Trichlorophenyl-benzoxime induces apoptosis in human colon carcinoma cells via activation of mitochondria dependent pathway

Purpose: To determine the apoptotic effect of trichlorophenyl-benzoxime (TCPB) on colorectal cancer (CRC) cells, and to elucidate the mechanism of action. Methods: Colon carcinoma cell lines (DLD-1 and HT-29) were used in this study. The cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin at 37 ˚C in an atmosphere of 5 % CO2 and 95 % air. When the cells attained 60 - 70 % confluency, they were treated with serum-free medium and graded concentrations of TCPB (1.0 – 6.0 μM) for 24 h. Cell viability and apoptosis were assessed using 3-(4, 5-dimethylthiazol-2-yl) 2, 5-diphenyltetrazolium bromide (MTT) and flow cytometric assays, respectively. Western blotting and 2', 7' dichlorofluorescein diacetate (DCFH DA) assays were used for the determination of expression levels of apoptotic proteins, and levels of reactive oxygen species (ROS), respectively. Results: Treatment of DLD-1 and HT-29 cells with TCPB led to significant and dose-dependent reductions in their viability, as well as significant and dose-dependent increases in the number of apoptotic cells (p < 0.05). Treatment of HT-29 cells with TCPB led to significant increases in the population of cells in the G0/G1 phase, but significant reduction of cell proportion in S and G2/M phases (p < 0.05). It also significantly and dose-dependently upregulated the expressions of caspase-3 and bax, down-regulation of the expression of bcl-2 (p < 0.05). TCPB treatment upregulated the expressions of p53, cytochrome c (cyt c), procaspase-3, and procaspase-9, but down-regulated the expression of pAkt dose-dependently (p < 0.05). The expression of Akt in HT-29 cells was not significantly affected by TCPB (p > 0.05). However, TCPB significantly enhanced the cleavage of PARP1, and significantly and dose-dependently increased the levels of ROS in HT-29 cells (p < 0.05). Conclusion: These results suggest that TCPB exerts apoptotic effect on CRC cells via activation of mitochondria-dependent pathway, and thus can be suitably developed for the management of colon cancer

RESEARCH ON THE RELATIONSHIP AMONG MARKET ORIENTATION, CUSTOMER RELATIONSHIP MANAGEMENT, CUSTOMER KNOWLEDGE MANAGEMENT AND BUSINESS PERFORMANCE

In the age of knowledge-based economy, business must effectively to improve its performance rely on implement customer relationship management and customer knowledge management. Market orientation is a kind of marketing logic which is focus on customers, it can help to construct the relationship and communicate knowledge with customers, in order to improve customer satisfaction, customer loyalty and gain and accumulate value for the business. This paper is intended to construct an integrated model. In this model it discusses the relationship between market orientation, customer relationship management and customer knowledge management, and their contribution to business performance. Key words: market orientation, customer knowledge management, customer relationship management; business performanc

Optimiranje podloge za uzgoj i sastava aminokiselina u miceliju gljive Pleurotus cornucopiae SD-01 obogaćenog kromom

Chromium(III) is an essential trace element for humans and animals. Pleutorus cornucopiae SD-01 is a nutritional and functional mushroom containing many kinds of bioactive ingredients. The aims of this work are to optimize the conditions of P. cornucopiae SD-01 cultivation with Cr enrichment in submerged culture by determining the dry cell mass, Cr content in mycelia and the rate of Cr enrichment, and to analyze the amino acid composition of Cr-enriched mycelia. The optimal medium contained (in g/L): potato 200, sucrose 25, yeast extract 4, KH2PO4 1 and MgSO4·7H2O 1. The optimum parameters of liquid culture were temperature 25 °C, cultivation time 6 days, the volume of the medium 100 mL, rotation speed 160 rpm and initial pH=6.5. Under the optimized conditions, the values of the dry cell mass, Cr content in mycelia and the rate of Cr enrichment were (6.63±0.35) g/L, (3670±211) μg/g and (12.15±1.01) % respectively, which were by (23.23±1.22), (18.19±1.06) and (45.68±2.67) % respectively, higher than those in the control. Chromium(III) in Cr-enriched mycelia was mainly combined with protein and polysaccharide. The contents of total amino acids and essential amino acids in Cr-enriched mycelia were increased by (31.25±0.58) and (44.26±0.76) %, respectively.Trovalentni je krom esencijalni element u tragovima, potreban za rast ljudi i životinja. Pleurotus cornucopiae SD-01 hranjiva je gljiva funkcionalnih svojstava, koja sadrži mnoge bioaktivne komponente. Svrha je ovoga rada bila optimirati uvjete uzgoja P. cornucopiae SD-01 u podlozi obogaćenoj kromom, i to određivanjem suhe stanične mase, udjela kroma u miceliju i stupnja obogaćivanja micelija, te analizirati aminokiselinski sastav micelija obogaćenog kromom. Optimalni sastav podloge bio je (u g/L): krumpir 200, saharoza 25, kvaščev ekstrakt 4, KH2PO4 1 i MgSO4·7H2O 1. Utvrđeni su optimalni parametri rasta: temperatura 25 °C, vrijeme uzgoja 6 dana, volumen podloge 100 mL, brzina rotacije 160 rpm i početni pH=6,5. U tim su uvjetima dobivene sljedeće vrijednosti: suha stanična masa od (6,63±0,35) g/L, veća za (23,23±1,22) %; udio kroma u miceliju od (3670±211) mg/g, veći za (18,19±1,06) % i stupanj obogaćivanja micelija kromom od (12,15±1,01) %, što je za (45,68±2,67) % više nego u kontrolnom uzorku. Utvrđeno je da se trovalentni krom u miceliju uglavnom veže za proteine i ugljikohidrate. Ukupni se udio aminokiselina u takvom obogaćenom miceliju povećao za (31,25±0,58) %, a udio esencijalnih aminokiselina za (44,26±0,76) %