The effect of long term exposure to copper on physiological condition and reproduction of sheep

The influence of copper upon some physiological parameters and reproduction in ewes was studied. Four groups of animals were investigated: 1/ control ewes (untreated); 2) ewes receiving copper as a supplement over the recommended amount of copper in food (10, 25 or 50 mg Cu/ewe/day); 3/ control, superovulated ewes; and 4) ewes treated with 50 mg copper during one month and then superovulated. After 10 months of daily exposure to 10 mg of copper/ewe/day no signs of toxicity on physiological condition and reproduction were found. In ewes exposed to 25 or 50 mg of copper a decrease in blood parameters and increase in concentration of Cu in blood and liver were noticed. The wavy pattern of follicles was disturbed and disorders in fecundity, prolificacy and pregnancy occurred. Significant differences between the number of corpora lutea in superovulated control animals and experimental (Cu 50 mg) ewes were observed.     

An enormously active and selective azapeptide inhibitor of cathepsin B.

The peptidomimetic Z-Arg-Leu-Arg-Agly-Ile-Val-OMe (where Agly means alpha-aza-glycyl, -NHNHCO-) is the strongest (K(i) = 480 pM) and the most selective inhibitor of cathepsin B to date, being approximately 2310 times as active to cathepsin B as to cathepsin K. In this paper we introduce the peptide and seek to rationalize its structure-activity relationships using molecular dynamics (MD) and NMR. It is shown that the -Agly-moiety restrains the peptide backbone to a bent shape, contrary to its parent peptide (with Gly in position 4), having its backbone extended and flexible. This fold is maintained in the plug covalently bound to the cathepsin B Cys29, in compliance with similar bends already observed in two other azapeptides attached to the active sites of cathepsin B. The MD simulation of the Z-Arg-Leu-Arg-Agly approximately cathepsin B complex suggests that, contrary to other potent inhibitors of cathepsin B, the current double Arg(1)/Arg(3) inhibitor, while maintaining the fold is able to form a unique ion cluster involving both Arg residues on the inhibitor part and two acidic Glu171 and Glu245 on the cathepsin B part, thus enhancing the affinity and subsequently the inhibiting power and selectivity of Z-Arg-Leu-Arg-Agly-Ile-Val-OMe to the observed extreme extent.     

FAME profiles in Pseudomonas vesicularis during catechol and phenol degradation in the presence of glucose as an additional carbon source.

The aim of this study was to evaluate the impact of catechol and phenol added to culture media separately and with glucose as an additional, easily-degradable carbon source on fatty acid methyl ester (FAME) composition in Pseudomonas vesicularis. Simultaneously, the degradation rates of aromatic substrates used were investigated in single and binary substrate systems. Both catechol and phenol treatments caused changes in the distribution of tested groups of fatty acids. The most noticeable changes included an increase in degree of fatty acid saturation, the appearance of branched and disappearance of hydroxy fatty acids as compared to the control sample with glucose. Under catechol or phenol treatment sat/unsat ratio showed the values of 8.63 and 11.38, respectively, whereas in control cells it reached the value of 2.66. The high level of saturation comes from the high content of cyclopropane fatty acids in bacteria under exposure to aromatic substrates, regardless of the presence of glucose. In these treatments their content was more than 3-fold higher compared to the control. It has been demonstrated that glucose supplementation of culture media containing single aromatic substrate extended the degradation rates of catechol and phenol by P. vesicularis, caused an increase in number of cells but did not significantly change the fatty acid profiles in comparison with bacteria growing on catechol and phenol added to the media individually.     

Effect of antisense peptide binding on the dimerization of human cystatin C--gel electrophoresis and molecular modeling studies

Human cystatin C (HCC) shows a tendency to dimerize. This process is particularly easy in the case of the L68Q HCC mutant and might lead to formation of amyloid deposits in brain arteries of young adults. Our purpose was to find ligands of monomeric HCC that can prevent its dimerization. Eleven antisense peptide ligands of monomeric HCC were designed and synthesized. The influence of these ligands on HCC dimerization was studied using gel electrophoresis and molecular modeling methods. The results suggest that all the designed peptides interact with monomeric HCC facilitating its dimerization rather than preventing it.     

Palindromes in proteins

Palindromes in DNA consist of nucleotides sequences that read the same from the 5'-end to the 3'-end, and its double helix is related by twofold axis. They occur in genomes of all organisms and have various functions. For example, restriction enzymes often recognize palindromic sequences of DNA. Palindromes in telomeres are crucial for initiation of replication. One can ask the questions, Do palindromes occur in protein, and if so, what function they play? We have searched the protein SWISSPROT database for palindromic sequences. A great number (26%) of different protein palindromes were found. One example of such protein is systemin, an 18-amino-acid-long peptide. It contains palindrome in its beta-sheet domain that interacts with palindromic fragment of DNA. The other palindrome containing protein is cellular human tumor suppressor p53. Oligonucleotide LTI-ITL has been observed in the crystal structure and is located close to a DNA recognizing domain. As the number of possible palindromic sequences of a given length is far much greater for proteins (20N) than for nucleic acids (4N), the study on their role seems to be an exciting challenge. Our results have clearly showed that palindromes are frequently occurring motives in proteins. Moreover, even very few examples that we have examined so far indicate the importance of further studies on protein palindromes.     

Enteropathogenic activity and invasion of HEp-2 cells by Aeromonas caviae clinical isolates

Twenty Aeromons caviae isolates from stool of children with diarrhea symptoms were examined for virulence-associated properties: production of cytotoxic and cytotonic toxins, and invasive ability. Most of A. caviae strains were cytotoxic to Vero and CHO cells and produced cytotonic toxins which caused elongation of CHO cells. Moreover, five of A. caviae strains revealed invasive ability towards HEp-2 cells.     

Polysome formation and stability in pea stem and root tissue

Polysome stability and the formation of various polysomal populations in pea stem and root tissue were examined. Both total ribosomal fraction and four polysome populations were isolated: FP (free polysomes), MBP (membrane-bound polysomes), CBP (cytoskeleton-bound polysomes) and CMBP (cytoskeleton-membrane-bound polysomes). The content of above mentioned populations decreased in roots and stems during germination. In both roots and stems a gradual decrease of FP participation in the total polysomal population was also observed during germination. On the other hand, an obvious increase in participation of CMBP population in the total polysomes pool was observed in later stages of germination. Increase of CMBP participation in pea root and stem tissues in later stages of germination is probably due to intensive enzymatic protein synthesis taking place in them. These proteins may participate in elongating growth of cells. The results of investigation on polysomes stability showed that total polysomes isolated from pea roots appeared to be more resistant to digestion by exogenous ribonuclease (EC 3.1.27.5) than polysomes isolated from stems. As protein-mRNA interactions are widely known and ribosomes are also very adhesive structures, numerous non-ribosomal proteins are present in the polysome preparations. We suppose that changes in proteins bound to polysomes indicated by us previously, significantly influence both the stability and also translatability of polysomes isolated from different plant organs.     

Induction of apoptosis by EGCG in selected tumour cell lines in vitro

One of the best recognised polyphenols of plant origin, epigallocatechin-3-gallate (EGCG) is contained mainly in green tea and in grapes. Studies performed in vivo and in vitro have demonstrated high probability of anti-neoplastic potential of the compound, due to its capacity to induce programmed cell death. The present studies were aimed at evaluation of apoptosis induction in cells of three selected tumour cell lines, subjected to action of various concentrations of EGCG. The experiment was performed on cultures of HEp-2 laryngeal carcinoma cells, LoVo colon carcinoma cells, HeLa cervical carcinoma cells and on normal myoepithelial cell line, HS. EGCG was found to induce apoptosis in cells of the examined neoplastic lines in a dose-related manner. Moreover, effect of EGCG on normal cells of HS line was found to be much less pronounced as compared to effects exerted on sensitive neoplastic cells.