Effects of 500-year mercury mining and milling on cancer incidence in the region of Idrija, Slovenia

The aim of the study was to determine whether the 500-year of mercury mining and milling in the Idrija region in Slovenia and the resulting environmental pollution with mercury and smelting wastes containing radon, has caused an increased cancer risk of the inhabitants. The polluted and the non-polluted parts of the region were defined. Cancer incidence from the two regions was compared. Cancer incidence among miners was investigated separately. In the polluted area male and female cancer incidence was higher than in the non-polluted area. Miners had an excess of incidence of total cancer, of oral and pharyngeal cancers and of lung cancer. As indicated by multivariate analysis the increased risk of miners could be assigned to their smoking and alcohol drinking habits. Higher estimated cumulative exposure to inorganic mercury seems to contribute to their risk as well. Most of the excess cancer incidence of the population from the polluted area could be explained by an unhealthy life style. In the case of lung cancer radon exposure contributes to the increased risk as well. Therefore, a well planed health promotion program and further sanitation of old houses is proposed.     

Influence of the "Rijeka model" of bioethics education on attitudes of medical students towards death and dying--a cross sectional study

The aim of this study was to assess attitude towards euthanasia, and the influence of socio-demographic data and death education carried out through the "Rijeka model" of bioethics education for the first-year medical students of the School of Medicine, University of Rijeka, Croatia. The cross-sectional study was conducted in the academic year 20031 2004. 124 (61% female) participants were surveyed by using an anonymous questionnaire before and after training. Catholics (p = 0.003) and students from areas with populations of less than 50,000 inhabitants (p = 0.001) had significantly negative attitude towards euthanasia than others before the course, yet no differences were found following this training. Attitude towards euthanasia was significantly positive after the course (p = 0.005). All items in the questionnaire, except "Croatia should legalise euthanasia", received more positive scores after the course. Death education carried through the "Rijeka model" of bioethics education has changed attitudes of medical students towards a more positive perception of euthanasia.     

Photocarcinogenesis--molecular mechanisms

The carcinogenicity (photocarcinogenicity) of sunlight to human skin has been recognized more than a century ago. Last decades numerous experimental studies show that UV rays damage DNA, cause gene mutations leading to the development of malignant tumors such basal cell carcinomas, squamous cell carcinomas and melanomas. The tumors occur most frequently in fair skinned people, and the mutations typically are found at dipyrimidine sites with C-T or / and CC-TT tandem double mutations. The authors briefly summarize their investigation of the p53 suppressor gene, and expose their hypothesis of hTERT involvement in cancerogenesis. Also their underline the importance of UV induced immunosuppression in photocarcinogenesis. Psoriatic patients are exposed to numerous cancerogens in their treatment. A better understanding of the mechanisms of photocarcinogenesis could provide new ways in the treatment of skin tumors.     

The potential impacts of changes in bear hunting policy for hunting organisations in Croatia

The brown bear (Ursus arctos) in Croatia is currently being managed through trophy hunting, with quotas allocated to local hunting organisations. Human–bear conflict is present at a low level, but any losses are compensated by the hunting organisations that benefit from bear hunting. Attitudes towards bears are generally positive, and the bear population appears stable, or even increasing. Croatia's current bear hunting policy relies upon both the ecological sustainability of the quotas and the economic sustainability of the hunting organisations. To address the first of these pillars of current policy, we used a two-sex matrix model of the bear population to investigate the biological sustainability of current hunting levels. The model suggests that if the annual allocated quota were fully realised, the population would suffer a considerable decrease over 10 years. A likely explanation for the mismatch between this result and the observed stability of the population is that the bear population size is underestimated. To address the second pillar, we quantified the current structure, costs and benefits of bear hunting to hunting organisations through an interview survey with hunting managers. We found that bear hunting is a substantial component of hunting organisations' income, supporting the other activities of the organisation. Croatia's recent accession to the EU will require changes in their bear management system, potentially stopping bear trophy hunting. Therefore, we assessed the changes in hunting organisations' budgets in the absence of bear hunting. Our results demonstrate that a loss of bear trophy hunting would result in a substantial loss of income to the hunting organisations. Moving bear hunting and compensation mechanisms from local management and responsibility to a more centralised system without trophy hunting, as suggested by EU legislation, will lead to considerable uncertainties. These include how to make centralised decisions on population targets and offtake levels for population control, given the uncertainty around population estimates, and on compensation payments given the loss of the current system which relies heavily on local income from trophy hunting, local relationships and informal monetary and non-monetary compensation.     

Antioxidant and Sensorial Properties of Linden Honey with Dried Apricots

The total phenol (TPh) and flavonoid contents (TFd), and antioxidant and sensorial properties of linden honey (LH) with dried apricots (20, 30, and 40%) were evaluated. TPh increased 4.3 times for LH40 (from 23.96 to 102.87 mg gallic acid equiv./100 g honey), while increase of TFd was slightly lower, ca. 2.9‐fold for LH40 (from 18.11 to 51.72 mg rutin equiv./100 g honey). Based on HPLC analysis, the most dominant phenolic compound was gallic acid (11.14 mg/100 g honey in LH and 42.65 mg/100 g honey in LH40). In three different assays, the antioxidant activity increased with increasing concentration of apricots in honey. The values varied from 13.36 for LH to 7.06 mg/ml for LH40; the values ranged from 189.83 for LH to 11.23 mg/ml for LH40; the RP_0.5 (reducing power) values ranged from 169.00 for LH to 27.60 mg/ml for LH40. Based on the correlation analysis, it is obvious that TPh and TFd were associated with the antioxidant activities of honey samples. A high degree of correlation existed between antioxidant activities of honey samples and TPh (R from 0.945 to 0.996) and TFd (R from 0.805 to 0.934). Obtained scores for individual sensory properties indicated very good quality of honey with dried apricots.     

Archaeal aminoacyl-tRNA synthetases interact with the ribosome to recycle tRNAs

Aminoacyl-tRNA synthetases (aaRS) are essential enzymes catalyzing the formation of aminoacyl-tRNAs, the immediate precursors for encoded peptides in ribosomal protein synthesis. Previous studies have suggested a link between tRNA aminoacylation and high-molecular-weight cellular complexes such as the cytoskeleton or ribosomes. However, the structural basis of these interactions and potential mechanistic implications are not well understood. To biochemically characterize these interactions we have used a system of two interacting archaeal aaRSs: an atypical methanogenic-type seryl-tRNA synthetase and an archaeal ArgRS. More specifically, we have shown by thermophoresis and surface plasmon resonance that these two aaRSs bind to the large ribosomal subunit with micromolar affinities. We have identified the L7/L12 stalk and the proteins located near the stalk base as the main sites for aaRS binding. Finally, we have performed a bioinformatics analysis of synonymous codons in the Methanothermobacter thermautotrophicus genome that supports a mechanism in which the deacylated tRNAs may be recharged by aaRSs bound to the ribosome and reused at the next occurrence of a codon encoding the same amino acid. These results suggest a mechanism of tRNA recycling in which aaRSs associate with the L7/L12 stalk region to recapture the tRNAs released from the preceding ribosome in polysomes.     

Role of Somatostatin Receptor-2 in Gentamicin-Induced Auditory Hair Cell Loss in the Mammalian Inner Ear

Hair cells and spiral ganglion neurons of the mammalian auditory system do not regenerate, and their loss leads to irreversible hearing loss. Aminoglycosides induce auditory hair cell death in vitro, and evidence suggests that phosphatidylinositol-3-kinase/Akt signaling opposes gentamicin toxicity via its downstream target, the protein kinase Akt. We previously demonstrated that somatostatin—a peptide with hormone/neurotransmitter properties—can protect hair cells from gentamicin-induced hair cell death in vitro, and that somatostatin receptors are expressed in the mammalian inner ear. However, it remains unknown how this protective effect is mediated. In the present study, we show a highly significant protective effect of octreotide (a drug that mimics and is more potent than somatostatin) on gentamicin-induced hair cell death, and increased Akt phosphorylation in octreotide-treated organ of Corti explants in vitro. Moreover, we demonstrate that somatostatin receptor-1 knockout mice overexpress somatostatin receptor-2 in the organ of Corti, and are less susceptible to gentamicin-induced hair cell loss than wild-type or somatostatin-1/somatostatin-2 double-knockout mice. Finally, we show that octreotide affects auditory hair cells, enhances spiral ganglion neurite number, and decreases spiral ganglion neurite length.     

Refined Candidate Region for F4ab/ac Enterotoxigenic Escherichia coli Susceptibility Situated Proximal to MUC13 in Pigs

F4 enterotoxigenic Escherichia coli (F4 ETEC) are an important cause of diarrhea in neonatal and newly-weaned pigs. Based on the predicted differential O-glycosylation patterns of the 2 MUC13 variants (MUC13A and MUC13B) in F4ac ETEC susceptible and F4ac ETEC resistant pigs, the MUC13 gene was recently proposed as the causal gene for F4ac ETEC susceptibility. Because the absence of MUC13 on Western blot from brush border membrane vesicles of F4ab/acR⁺ pigs and the absence of F4ac attachment to immunoprecipitated MUC13 could not support this hypothesis, a new GWAS study was performed using 52 non-adhesive and 68 strong adhesive pigs for F4ab/ac ETEC originating from 5 Belgian farms. A refined candidate region (chr13: 144,810,100–144,993,222) for F4ab/ac ETEC susceptibility was identified with MUC13 adjacent to the distal part of the region. This candidate region lacks annotated genes and contains a sequence gap based on the sequence of the porcine GenomeBuild 10.2. We hypothesize that a porcine orphan gene or trans-acting element present in the identified candidate region has an effect on the glycosylation of F4 binding proteins and therefore determines the F4ab/ac ETEC susceptibility in pigs.     

Converting the Yeast Arginine Can1 Permease to a Lysine Permease

Amino acid uptake in yeast cells is mediated by about 16 plasma membrane permeases, most of which belong to the amino acid-polyamine-organocation (APC) transporter family. These proteins display various substrate specificity ranges. For instance, the general amino acid permease Gap1 transports all amino acids, whereas Can1 and Lyp1 catalyze specific uptake of arginine and lysine, respectively. Although Can1 and Lyp1 have different narrow substrate specificities, they are close homologs. Here we investigated the molecular rules determining the substrate specificity of the H⁺-driven arginine-specific permease Can1. Using a Can1-Lyp1 sequence alignment as a guideline and a three-dimensional Can1 structural model based on the crystal structure of the bacterial APC family arginine/agmatine antiporter, we introduced amino acid substitutions liable to alter Can1 substrate specificity. We show that the single substitution T456S results in a Can1 variant transporting lysine in addition to arginine and that the combined substitutions T456S and S176N convert Can1 to a Lyp1-like permease. Replacement of a highly conserved glutamate in the Can1 binding site leads to variants (E184Q and E184A) incapable of any amino acid transport, pointing to a potential role for this glutamate in H⁺ coupling. Measurements of the kinetic parameters of arginine and lysine uptake by the wild-type and mutant Can1 permeases, together with docking calculations for each amino acid in their binding site, suggest a model in which residues at positions 176 and 456 confer substrate selectivity at the ligand-binding stage and/or in the course of conformational changes required for transport.     

Different Incubation Times of Cells After Gene Electrotransfer in Fetal Bovine Serum Affect Cell Viability,but Not Transfection Efficiency

Electroporation as a delivery method is increasingly important in gene therapy, not only in vivo but also in in vitro experimental systems. Different applications of gene electrotransfer require high viability of cells and high transfection efficiency of gene electrotransfer. It was already demonstrated that the addition of fetal bovine serum (FBS) immediately after gene electrotransfer leads to improved cell survival and transfection efficiency. Therefore, the aim of the study was to determine whether prolonged incubation of cells in FBS, for more than standard 5 min, can lead to increased transfection efficiency and improved cell survival. Different murine melanoma and murine and human endothelial cell lines were transfected with plasmid encoding green fluorescent protein and then incubated for different periods of time in FBS (5–30 min). Transfection efficiency was determined by flow cytometry and fluorescence microscopy and cell survival by cell viability assay. Prolonged incubation of cells in FBS after gene electrotransfer had varying effect on cell survival, which was decreased in melanoma cell lines B16F1 and B16F10, minimally affected in SVEC4-10 and HUVEC cells and increased in 2H11 cell at 30 min of incubation time in FBS. On the other hand, transfection efficiency of gene electrotransfer was not affected by long incubation of cell in FBS, regardless of the cell line used. The results of our study emphasize the importance of optimization of gene electrotransfer protocol for particular cells and specific purposes of gene electrotransfer, taking into account the importance of transfection efficiency and cell survival.