Katarina Caput Mihalić

PSEUDO-NITZSCHIA BLOOMS IN THE ZRMANJA RIVER ESTUARY (EASTERN ADRIATIC SEA)

The spatial and temporal distribution of the diatom genus Pseudo-nitzschia was investigated in the small, highly stratified Zrmanja River estuary (eastern Adriatic Sea) from 1998 to 2004. During this period, Pseudo-nitzschia spp. mostly dominated the phytoplankton assemblages (abundance close to 1.3 × 106 cells L−1, appeared in 51% of the samples). The maximum abundance of Pseudo-nitzschia spp. was during mid-winter (February 1999), when it contributed up to 85% of the total diatom abundance. Pseudo-nitzschia bloom was significantly correlated with phosphorous concentration and temperature, whereas there was no significant correlation with total inorganic nitrogen, silicate and salinity. The very broad range of environmental factors over which Pseudo-nitzschia spp. were observed suggests there was more than one Pseudo-nitzschia species in the diatom assemblage. Morphological analysis has shown that the dominating Pseudo-nitzschia species corresponded to Pseudo-nitzschia calliantha. Two other species were encountered; one identified as Pseudo-nitzschia heimii, while other has yet to be identified. This study was the first to find P. heimii in the Adriatic Sea and the first to record the presence of a potentially toxic diatom P. calliantha in an Adriatic estuary.

The occurrence and ecology of the centric diatom Cyclotella choctawhatcheeana Prasad in a Croatian estuary

The centric diatom Cyclotella choctawhatcheeana Prasad has been found for the first time in the Adriatic Sea. It develops dense populations in the highly stratified, oligotrophic, karstic estuary of the Zrmanja River (eastern Adriatic Sea, Croatia). Cyclotella choctawhacheeana contributed to the phytoplankton community in the estuary with high abundance of up to 2 x 10(6) cells L-1. Maximum abundance was found in the marine layer, below the halocline, at a salinity range between 13-28, but it was able to develop over a very wide salinity range. The maximum development appeared in the temperature range of 8-18 degrees C. It was present in 52% of samples throughout the year. The significant negative correlation with silica, nitrates and total inorganic nitrogen showed its preference for a marine, oligotrophic environment. We consider C. choctawhacheeana to be a significant constituent of the phytoplankton of the east Adriatic coast. It can be assumed that its succes is based on species wide tolerance to fluctuations of environmental factors.

Cytotoxic Activity of Novel Palladium-Based Compounds on Leukemia Cell Lines

Effective treatment methods for human leukemia are under development, but so far none of them have been found to be completely satisfactory. It was recently reported that palladium complexes have significant anticancer activity as well as lower toxicity compared with some clinically used chemotherapeutics. The anticancer activities of two novel palladium(II) complexes, [Pd(sac)(terpy)](sac)·4H2O and [PdCl(terpy)](sac)·2H2O, were tested against three human leukemia cell lines, Jurkat, MOLT-4, and THP-1, in comparison with cisplatin and adriamycin. The cytotoxic effect of the drugs was determined using the MTT assay. Cell death was assessed using fluorescein isothiocyanate-annexin/propidium iodide staining for flow cytometry. Furthermore, p53 phosphorylation, poly(ADP-ribose) polymerase cleavage, and Bax and Bcl-2 mRNA levels were examined to elucidate the mechanism of cell death induction. Both complexes exhibited a significant dose-dependent antigrowth effect in vitro. The complexes predominately induced apoptosis, but necrosis was also observed. In-vitro results have shown that palladium(II) complexes may be regarded as potential anticancer agents for treating human leukemia. Therefore, further analysis to determine the putative mechanism of action and in-vivo studies on animal models are warranted.

Human Mesenchymal Stem Cells Differentiation Regulated by Hydroxyapatite Content within Chitosan-Based Scaffolds under Perfusion Conditions

The extensive need for hard tissue substituent greatly motivates development of suitable allogeneic grafts for therapeutic recreation. Different calcium phosphate phases have been accepted as scaffold’s components with positive influence on osteoinduction and differentiation of human mesenchymal stem cells, in terms of their higher fraction within the graft. Nevertheless, the creation of unlimited nutrients diffusion through newly formed grafts is of great importance. The media flow accomplished by perfusion forces can provide physicochemical, and also, biomechanical stimuli for three-dimensional bone-construct growth. In the present study, the influence of a different scaffold’s composition on the human mesenchymal stem cells (hMSCs) differentiation performed in a U-CUP bioreactor under perfusion conditioning was investigated. The histological and immunohistochemical analysis of cultured bony tissues, and the evaluation of osteogenic genes’ expression indicate that the lower fraction of in situ formed hydroxyapatite in the range of 10–30% within chitosan scaffold could be preferable for bone-construct development.