Comparison of in vitro antileukemic activity of obatoclax and ABT-737

Obatoclax and ABT-737 belong to a new class of anticancer agents known as BH3-mimetics. These agents antagonize the anti-apoptotic members of Bcl-2 family. The Bcl-2 proteins modulate sensitivity of many types of cancer cells to chemotherapy. Therefore, the objective of the present study was to examine and compare the antileukemic activity of obatoclax and ABT-737 applied alone, and in combination with anticancer agent, mafosfamide and daunorubicin. The in vitro cytotoxic effects of the tested agents on human leukemia cells were determined using the spectrophotometric MTT test, Coulter electrical impedance method, flow cytometry annexin V–fluorescein/propidium iodide assay, and light microscopy technique. The combination index analysis was used to quantify the extent of agent interactions. BH3 mimetics significantly decreased the leukemia cell viability and synergistically enhanced the cytotoxic effects induced by mafosfamide and daunorubicin. Obatoclax affected the cell viability to a greater degree than did ABT-737. In addition, various patterns of temporary changes in the cell volume and count, and in the frequency of leukemia cells undergoing apoptosis, were found 24 and 48 h after the tested agent application. ABT-737 combined with anticancer agents induced apoptosis more effectively than obatoclax when given in the same combination regimen. The results of the present study point to the different antileukemic activities of obatoclax and ABT-737, when applied alone, and in combination with anticancer agents. A better understanding of the exact mechanisms of BH3 mimetic action is of key importance for their optional use in cancer therapy

POJAVA GENETSKI MODIFICIRANIH USJEVA U KRMIVIMA U POLJSKOJ

The aim of this study was to detect, identify, and quantify genetically modified (GM) crops by DNA analyses in feed used for animal feeding in Poland. This approach was based on PCR and Real Time PCR techniques and was applied to official genetically modified feed control in the National Veterinary Research Institute in Pulawy. Within six years studies (2004-2009) in 231 (66%) samples, out of 350 samples of compound feed and feed materials, genetically modified soybean, maize or rapeseed were found. The highest GMO content was found in soybean meal samples and compound feed which consisted of soybean meal. Among 135 analyzed soybean meal samples in 127 (94%) of them GM soya GTS 40-3-2 was present. As regards compound feed samples, GM soya was identified in 93 (83%) samples out of all 112 samples. The presence of GM maize was rare. Only in 11 (11%) samples, out of all 98 samples of maize, GM maize varieties were found (10 x MON810, 2 x Bt11, 2 x T25). Moreover analyses of 112 compound feed samples indicated that, 16 of them contained Mon810, 4 samples Bt11 and 3 samples T25 GM maize varieties. As regards rapeseed analysis in 5 checked rapeseed samples there were no GM varieties. Another study focused only on GM rapeseed, provided for us independently by official feed control, proved previous results. Out of 100 samples only in 2 of them LibertylinkTM rape was present in the amount below limit of quantification (LOQ) of our method. Achieved results of feed samples analyses indicate that the use of GM soya is common in animal feeding in Poland as main source of protein. The usage of GM maize is low, but mainly MON810 maize was found this variety is grown in Europe. As regards rapeseed there were very few GM rape seed positive samples.Cilj ovog rada bio je otkriti, identificirati i kvantificirati genetski modificirane (GM) usjeve DNK analizama u hrani za životinje u Poljskoj. Pristup se temeljio na tehnikama PCR i Real Time, što se primjenjuju u službenoj kontroli genetski modificirane hrane u Državnom veterinarskom istraživačkom institutu u Pulawyju. U šest godina istraživanja (2004-2009) na 211 (60%) uzoraka, od 350 uzoraka krmnih smjesa i krmnog materijala, pronađeni su genetski modificirana soja, kukuruz i uljna repica. Najviši sadržaj GMO-a nađen je u uzorcima sojinog brašna i krmnih smjesa koje su se sastojale od sojinog brašna. Među 135 analiziranih uzoraka sojinog brašna u 127 (94%) nalazio se GM soje GTS 40-3-2. Što se tiče uzoraka krmnih smjesa u 93 (83%) uzorka GM soje identificiran je u svih 112 uzoraka. Prisutnost GM kukuruza bila je rijetka. Samo u 11 (!!%) od svih 98 uzoraka kukuruza nađene su vrste GM (10 x MON810, 2 x Bt11, 2 x T25). Osim toga analize 112 uzoraka krmnih smjesa pokazale su da ih je 16 sadržavalo Mon810, 4 uzorka Bt11 i 3 uzorka T25 vrste kukuruza. Što se tiče analize uljne repice u 5 pregledanih uzoraka nije bilo GM vrsta. Druge analize usmjerene samo na GM uljne repice, što smo ih obavili neovisno o službenoj kontroli hrane potvrdile su gornje rezultate. U 100 uzoraka samo u 2 uzorka nađen je Libertylink TM repice u iznosu ispod granice kvantifikacije (LOQ) naše metode. Dobiveni rezultati analiza uzoraka krmiva pokazuju da je upotreba GM soje uobičajena u hranidbi životinja u Poljskoj kao glavni izvor bjelančevina. Upotreba GM kukuruza je sada na niskoj razini, a nađen je uglavnom kukuruz MON810 zbog uzgoja te vrste u Europi. Što se tiče uljne repice bilo je vrlo malo pozitivnih GM uzoraka

In vitro response of human pathological hematopoietic cells to fludarabine phosphate

The present study was undertaken to determine a possible influence of fludarabine (fludarabine phosphate, F-ara-AMP) on the cell viability and count. The experiments were performed in vitro on human acute lymphoblastic MOLT-4 cells, human acute myeloblastic ML-1 cells, and human histiocytic lymphoma U-937 cells. The research was conducted using the spectrophotometric and Beckman Coulter methods. The cell viability was analyzed using MTT assay. The cell count was detected using an electronic Z2 Coulter counter. Temporary changes in the cell viability and count were assessed at 24h and 48h after F-ara-AMP application. The in vitro activity of fludarabine phosphate against MOLT-4, ML-1, and U-937 cells was compared. F-ara-AMP applied at the four concentrations - 250 nM, 500 nM, 750 nM, and 1 μM - distinctly decreased the viability and count of the pathological hematopoietic cells. The effects of F-ara-AMP on MOLT-4, ML-1, and U-937 cells were dependent on the tested agent and its dose, the time intervals after the agent application, and the cell line used. ML-1 and U-937 cells appeared to be more resistant than MOLT-4 cells to the action of fludarabine phosphate. The in vitro response of the three human pathological hematopoietic cell lines to the F-ara-AMP action, was shown

A Case of Extensive Infl ammatory Changes (Osteomyelitis) in an Infant’s Skeleton from the Medieval Burial Ground (11th–12th c) in Wawrzeńczyce (Near Krakow)

The aim of this study was to diagnose and describe extensive infl ammatory changes in a child’s skeleton from Wawrzeńczyce, (the medieval period). The aim of the analysis was to determine the nature of the infl ammatory changes and their etiology by means of macroscopic techniques as well as X-ray analysis. The tests revealed that the individual suffered from a hematogenous multifocal osteitis. This condition might have been a result of an acute or sub-acute osteitis, and the untreated form of osteomyelitis might have contributed to the infection of the entire developing organism, leading to death