Pregled tradicionalnih pojmovanj in kritični pretres najnovejših teorij zahtevata, da se periodiziranje slovenske romantike uskladi z novimi vidiki evropske literarne vede in posebnimi razvojnimi ...značilnostmi romantike pri slovanskih narodih: do leta 1830 pr Slovencih še ni mogoče razpravljati o romantiki, ampak šele o predromantiki; predromantika se razmahne po letu 1809; romantika traja do leta 1848, sledita ji postromantika in predrealizem.
Proteolytic enzymes have been proposed as new biological prognostic indicators to facilitate decisions about treatment of breast cancer patients following surgery. We reported earlier that the ...activities of cysteine proteinases (CP), cathepsin (Cat) B and cathepsin (Cat) L and the expression of stefin A might be associated with breast tumor progression and prognosis. Here, the protein concentrations of Cats D, B and L and stefin A have been measured in a series of 60 matched pairs of breast tumours and control adjacent tissues, using ELISAs developed in our laboratory. Median tumor concentrations of Cat D (47 pm/mg), Cat B (222 ng/mg) and Cat L (88 ng/mg) were significantly (p<0.0005) increased by 7 fold, 27 fold and 6 fold, respectively. Much greater increases in the activities of Cat B (63 fold) and of Cat L (274 fold) were found, indicating activation of proCat B and proCat L and/or to a decrease in specific endogenous cystatins. However, the 1.6-fold decreased (p<0.0001) levels of inhibition by cystatins could not be entirely responsible for more than 100-fold increased ratio of CP:cystatins activity. Moreover, stefin A was either increased or decreased in tumor samples, resulting in a 1.4-fold median increase in tumors. Comparing the biological parameters with the established histo-pathological prognosticators, we found that the increased protein concentration of Cat B was associated with lymph node involvement (p<0.009) and higher stage (p<0.003), and both Cat B and Cat L activities were more increased in high grade tumours (p<0.05). Survival analysis revealed that stefin A was the most significant prognostic factor for disease-free (p<0.008) and overall survival (p<0.02), followed by increased Cat B activity and protein concentration. Cat L was of borderline significance while Cat D was not significant for prognosis. We conclude that enhanced activation of CP, due partially to an imbalance between cysteine proteinases and inhibitors is linked to the progression of breast cancer. Larger sample size is needed to confirm the prognostic significance of stefin A, Cat B and Cat L.
Kinetic stopped-flow measurements of refolding of the recombinant sulphonated procathepsin S from 6 M urea are presented. The experiments were performed using intrinsic tryptophan fluorescence and ...fluorescence of the hydrophobic probe 1-anilino-naphthalene-8-sulfonate (ANS). Initially, (
t
1/2=3±1 ms) an intermediate with increased ANS fluorescence and protected tryptophan environment is formed. Much later, a slow increase in ANS fluorescence occurs with no accompanying changes in tryptophan fluorescence. The reaction of the slow ANS fluorescence increase correlates with the rate of aggregation as shown by the size exclusion chromatography (SEC). For comparison, the folding reactions of the reduced chicken cystatin were measured, both, by intrinsic tryptophan and extrinsic ANS fluorescence. An early intermediate forms very fast in the refolding of reduced chicken cystatin on 6-fold dilution from 5.7 M GuHCl (
t
1/2=5±2 ms), similarly to that observed for the sulphonated procathepsin S. ANS fluorescence and tryptophan fluorescence decrease further (
t
1/2=100±50 ms) leading to a late, `more structured' intermediate which is prone to dimerization.
Kad je Anton Vodnik pripremao antologiju pod naslovom Slovenačka savremena lirika, koja je objavljena 1933. godine, uvrstio je u nju većinom pesme nastale između 1900. i 1930. godine. Pola veka ...kasnije, nama se pojam savremene slovenačke lirike, kakav u to vreme postoji, čini manje ili više identičan sa lirskom poezijom u poslednjih 30 godina, tj. od 1950. do 1980. Tako vremenski označena, »savremenost« je svakako tačnije određena, godine je istorijski omeđuju, raščlanjuju iznutra i spolja, možda je čak i zasnivaju. Tako se savremena slovenačka lirika hoćeš-nećeš ispoljava kao vremenski ograničeni period i zauzima posebno mesto u razvoju dosadašnje slovenačke poezije; razlikuje se i od onog što je bilo pre nje, ali i od onog što će doći posle nje.
A synthetic gene coding for the cysteine proteinase inhibitor (desSer 1 Ile29 Leu89) chicken cystatin was cloned and expressed in E. coli. The gene was assembled from 12 oligonucleotides and inserted ...into vector pUC 8. Expression as fusion protein was performed in a temperature‐inducible E. coli system. The expression product was synthesized as 20% of total E. coli protein. The fusion protein was purified, the chicken cystatin homologue was split off with CNBr and the N‐terminal sequence confirmed up to position 37. The properties of the purified material correspond to those of natural chicken cystatin. The recombinant cystatin variant binds anti‐chicken cystatin IgG, is inhibitorily active and displays K
i values with papain and with cathepsin B similar to those determined for natural chicken cystatin.
Cysteine cathepsins are lysosomal cysteine proteases that are involved in a number of important biological processes, including intracellular protein turnover, propeptide and prohormone processing, ...apoptosis, bone remodelling, and reproduction. In cancer, the cathepsins have been linked to extracellular matrix remodelling and to the promotion of tumour cell motility, invasion, angiogenesis and metastasis, resulting in poor outcome of the disease. The levels of cathepsins as well as of their endogenous inhibitor cystatins in clinical samples have been suggested as potential biomarkers, and this chapter is focused on their role to predict the diagnosis, risk of recurrence and death, and response to therapy in patients with cancer.
We have investigated the steady state kinetics of the degradation of native fibrillar collagen at pH 3.4 by four collagenolytic cathepsins of rabbit spleen. For each enzyme, the dependence of initial ...velocity on collagen concentration was well described by the Michaelis-Menten mechanism. Km, expressed as the concentration of triple-helical chains, and kcat values were determined for cathepsins B, L, N and S. The ratio of Kcat to Km suggest that cathepsins L and N are far more effective at collagen solubilization than either cathepsins S or B. Ki values were determined for the inhibition of collagenolytic activity at pH 3.4 using cystatin, a naturally-occurring cysteine proteinase inhibitor. All four cysteine proteinases were inhibited by cystatin in this assay system, although it was found to be a tighter binding inhibitor of cathepsin L, than for cathepsins N and S (approximately 5-fold less), or cathepsin B (approximately 500-fold less).