Platinum(ii/iv) complexes containing ethylenediamine-N,N'-di-2/3-propionate ester ligands induced caspase-dependent apoptosis in cisplatin-resistant colon cancer cells

Several new R(2)eddp (R = i-Pr, i-Bu; eddp = ethylenediamine-N,N'-di-3-propionate) esters and corresponding platinum(ii) and platinum(iv) complexes of the general formula [PtCl(n)(R(2)edda-type)] (n = 2, 4) were synthesized and characterized by spectroscopic methods (IR, (1)H and (13)C NMR) and elemental analysis. The crystal structure of platinum(iv) complex [PtCl(4){(c-Pe)(2)eddip}] () was resolved and is given herein. Ligand precursors, platinum(ii), and platinum(iv) complexes were tested against eight tumor cell lines (CT26CL25, HTC116, SW620, PC3, LNCaP, U251, A375, and B16). Selectivity in the action of those compounds between tumor and two normal primary cells (fibroblasts and keratinocytes) are discussed. A structure-activity relationship of these compounds is discussed. Furthermore, cell cycle distribution, induction of necrosis, apoptosis, autophagy, anoikis, caspase activation, ROS, and RNS are presented on the cisplatin-resistant colon carcinoma HCT116 cell line.     

Interactions of quercetin and its lanthane complex with double stranded DNA/RNA and single stranded RNA: spectrophotometric sensing of poly G

Spectrophotometric titrations revealed that stability of the quercetin/double stranded (ds) DNA or double stranded (ds) RNA non-covalent complexes is significantly higher compared to the quercetin/ss-RNA complexes. This observation can easily be correlated with the significantly larger aromatic surface of base pairs compared to single nucleobases, and it is in good agreement with other experimental data pointing toward intercalative binding mode of quercetin. Fluorescence increase of quercetin induced by ds-RNA is significantly stronger than observed for ds-DNA, offering usage of quercetin as the ds-RNA selective fluorescent probe. Also, addition of poly G yielded more than order of magnitude stronger changes in UV/visible and fluorescence spectrum of quercetin compared to the changes upon addition of poly A and poly U revealing possible usage of quercetin as a powerful spectroscopic probe for poly G sequences. Stability and stoichiometry of lanthane(III)/quercetin complexes in physiologically relevant aqueous media was determined. The interactions of (LaQ)(3+) with double stranded DNA and RNA were significantly different compared to the free quercetin, revealing increase of complex stability and thus significant impact of La(III) in binding of (LaQ)(3+) to polynucleotides. Similar results were observed for interactions of (LaQ)(3+) with single stranded RNA.     

Developmental patterns of caspase-3, bax and bcl-2 proteins expression in the human spinal ganglia

The distribution of the bcl-2, bax and caspase-3 proteins was investigated in the cells of developing human spinal ganglia. Paraffin sections of 10 human conceptuses between 5th and 9th gestational weeks were analysed morphologically, immunohistochemically and by TUNEL-method. Cells positive to caspase-3 had brown stained nuclei or nuclear fragmentations. At earliest stages, 6% of ganglion population were caspase-3 positive cells. Later on, a significant increase in number of caspase-3 positive cells appeared, particularly in the ventral part of ganglia (12%), and subsequently decreased to 6%. TUNEL-positive cells had the same distribution pattern as caspase-3 positive cells. Bax-positive cells followed the developmental pattern similar to caspase-3 cells, changing in range between 20% and 32%. There were 8% of bcl-2 positive cells at earliest stages. They increased significantly in dorsal part of the ganglion during the 7th week (28%), and than dropped to 15% by the end of the 8th week. These findings suggest a ventro-dorsal course of development in human spinal ganglia. Number of bcl-2, bax and caspase-3 positive cells changed in a temporally and spatially restricted manner, coincidently with ganglion differentiation. While apoptosis might control cell number, bcl-2 could act in suppression of apoptosis and enhancement of cell differentiation.     

Multiple nutrients limit litterfall and decomposition in a tropical forest

To explore the importance of 12 elements in litter production and decomposition, we fertilized 36 1600 m²-plots with combinations of N, P, K, or micronutrients (i.e. B, Ca, Cu, Fe, Mg, Mn, Mo, S, Zn) for 6 years in a lowland Panamanian forest. The 90% of litter falling as leaves and twigs failed to increase with fertilization, but reproductive litter (fruits and flowers) increased by 43% with N. K enhanced cellulose decomposition; one or more micronutrients enhanced leaf-litter decomposition; P enhanced both. Our results suggest tropical forests are a non-Liebig world of multiple nutrient limitations, with at least four elements shaping rates of litterfall and decomposition. Multiple metallomic enzymes and cofactors likely create gradients in the break down of leaf litter. Selection favours individuals that make more propagules, and even in an N-rich forest, N is a non-substitutable resource for reproduction.     

Highly sulfated glycosaminoglycans inhibit aggrecanase degradation of aggrecan by bovine articular cartilage explant cultures.

The catabolism of 35S-labeled aggrecan and loss of tissue glycosaminoglycans was investigated using bovine articular cartilage explant cultures maintained in medium containing 10(-6) M retinoic acid or 40 ng/ml recombinant human interleukin-1alpha (rHuIL-1alpha) and varying concentrations (1-1000 microg/ml) of sulfated glycosaminoglycans (heparin, heparan sulfate, chondroitin 4-sulfate, chondroitin 6-sulfate, dermatan sulfate and keratan sulfate) and calcium pentosan polysulfate (10 microg/ml). In addition, the effect of the sulfated glycosaminoglycans and calcium pentosan polysulfate on the degradation of aggrecan by soluble aggrecanase activity present in conditioned medium was investigated. The degradation of 35S-labeled aggrecan and reduction in tissue levels of aggrecan by articular cartilage explant cultures stimulated with retinoic acid or rHuIL-1alpha was inhibited by heparin and heparan sulfate in a dose-dependent manner and by calcium pentosan polysulfate. In contrast, chondroitin 4-sulfate, chondroitin 6-sulfate, dermatan sulfate and keratan sulfate did not inhibit the degradation of 35S-labeled aggrecan nor suppress the reduction in tissue levels of aggrecan by explant cultures of articular cartilage. Heparin, heparan sulfate and calcium pentosan polysulfate did not adversely affect chondrocyte metabolism as measured by lactate production, incorporation of [35S]-sulfate or [3H]-serine into macromolecules by articular cartilage explant cultures. Furthermore, heparin, heparan sulfate and calcium pentosan polysulfate inhibited the proteolytic degradation of aggrecan by soluble aggrecanase activity. These results suggest that highly sulfated glycosaminoglycans have the potential to influence aggrecan catabolism in articular cartilage and this effect occurs in part through direct inhibition of aggrecanase activity.     

Gelatinous invertebrate zooplankton of the South Adriatic: species composition and vertical distribution

The species composition and vertical distribution of gelatinousinvertebrate zooplankton were investigated from April 1993 toJune 1994 in the 0–1000 m water column at a deep-sea stationin the northern part of the South Adriatic Pit. Fifty-sevenspecies were identified: 11 hydromedusae, 13 calycophores, 3ctenophores, 3 heteropods, 10 pteropods, 8 polychaetes and 9chaetognaths. The pteropod Desmopterus papilio and the heteropodProtatlanta mediterranea were recorded for the first time inthe Adriatic Sea. Data from this study differed from those ofprevious investigations in the South Adriatic as regards thenumerically dominant polychaete and pteropod species. All investigatedgroups generally were more abundant in the upper 100 m and decreasedwith depth. Different vertical distributions of life stageswere observed for those species that occupy a wide depth range:Persa incolorata, Solmissus albescens, Limacina inflata, Cymbuliaperoni, Pelagobia longicirrata, Sagitta lyra and Sagitta decipiens.     

A Lack of Immune System Genes Causes Loss in High Frequency Hearing but Does Not Disrupt Cochlear Synapse Maturation in Mice

Early cochlear development is marked by an exuberant outgrowth of neurites that innervate multiple targets. The establishment of mature cochlear neural circuits is, however, dependent on the pruning of inappropriate axons and synaptic connections. Such refinement also occurs in the central nervous system (CNS), and recently, genes ordinarily associated with immune and inflammatory processes have been shown to play roles in synaptic pruning in the brain. These molecules include the major histocompatibility complex class I (MHCI) genes, H2-K^b and H2-D^b, and the complement cascade gene, C1qa. Since the mechanisms involved in synaptic refinement in the cochlea are not well understood, we investigated whether these immune system genes may be involved in this process and whether they are required for normal hearing function. Here we report that these genes are not necessary for normal synapse formation and refinement in the mouse cochlea. We further demonstrate that C1qa expression is not necessary for normal hearing in mice but the lack of expression of H2-K^b and H2-D^b causes hearing impairment. These data underscore the importance of the highly polymorphic family of MHCI genes in hearing in mice and also suggest that factors and mechanisms regulating synaptic refinement in the cochlea may be distinct from those in the CNS.