Efficient apoptosis and necrosis induction by proteasome inhibitor: bortezomib in the DLD-1 human colon cancer cell line

The inhibition of the 26S proteasome evokes endoplasmic reticulum stress, which has been shown to be implicated in the antitumoral effects of proteasome inhibitors. The cellular and molecular effects of the proteasome inhibitor—bortezomib—on human colon cancer cells are as yet poorly characterized. Bortezomib selectively induces apoptosis in some cancer cells. However, the nature of its selectivity remains unknown. Previously, we demonstrated that, in contrast to normal fibroblasts, bortezomib treatment evoked strong effect on apoptosis of breast cancer cells incubated in hypoxic and normoxic conditions. The study presented here provides novel information on the cellular effects of bortezomib in DLD-1 colon cancer cells line. We observe twofold higher percentage of apoptotic cells incubated for 48 h with 25 and 50 nmol/l of bortezomib in hypoxic conditions and four-, fivefold increase in normoxic conditions in comparison to control cells, incubated without bortezomib. It is of interest that bortezomib evokes strong effect on necrosis of DLD-1 colon cancer cell line. We observe the sixfold increase in necrosis of DLD-1 cells incubated with 25 or 50 nmol/l of bortezomib for 48 h in hypoxia and fourfold increase in normoxic conditions in comparison to adequate controls. We suggest that bortezomib may be candidates for further evaluation as chemotherapeutic agents for human colon cancer.     

Ethylene and IAA interactions in the inhibition of photoperiodic flower induction of <Emphasis Type="Italic">Pharbitis nil</Emphasis>

It has been shown that both IAA and ethylene application inhibit flower induction in the short-day plant Pharbitis nil. However application of IAA has elevated ethylene production in this plant, as well. Strong enhancement of ethylene production is also correlated with the night-break effect, which completely inhibits flowering. In order to determine what the role of IAA and ethylene is in the photoperiodic flower induction in Pharbitis nil, we measured changes in their levels during inductive and non-inductive photoperiods, and the effects of ethylene biosynthesis and action inhibitors on inhibition of flowering by IAA. Our results have shown that the inhibitory effect of IAA on Pharbitis nil flowering is not physiological but is connected with its effect on ethylene biosynthesis.     

Velocity distribution of the anterograde and retrograde transport of extracellular particles byAmoeba proteus

Summary The transverse velocity profiles of the anterograde flow of particles on the cell surface and around it are approximately parabolic. The peak velocity is recorded close to the membrane and the descendent arm of the profile is viscosity-dependent. It indicates that the extracellular forward flow is probably generated by a forward movement of the fluid fraction of the membrane itself. The retrograde component of extracellular movements is manifested by particles kept on the cell surface by adhesion, which behave exactly as the ectoplasmic layer on the opposite side of the membrane,i.e., they probably reflect the movement of that fraction of the surface material which is attached to the cortical microfilaments. In the longitudinal profile, the velocity of anterograde flow rises from the tail to the front of amoeba, but is generally related to the effective cell locomotion rate and not to the movements of any intracellular layer. Around the cells deprived of any attachment to the substratum, which cannot locomote but manifest vigorous intracellular movements, the anterograde flow ceases at least along 2/3 of their lenght. It persists, however, around the frontal fountain zone, where other particles still move backwards together with the retracted ectoplasmic layer. This indicates that the role of the forward flow of and on the cell surface is to compensate for: (1) the increase of the surface area in the frontal regions due to locomotion, (2) the withdrawal of a part of material which is hauled back by the retracting cortical layer. A comprehensive scheme of the velocity distribution within the different layers of a moving amoeba and around it has been constructed on the basis of present and earlier data.Study supported by the Research Project CPBP 04.01 of the Polish Academy of Science.I dedicate this paper to Professor K. E. Wohlfarth-Bottermann with the best wishes for his 65th birthday.     

Effects of tin and lead on organ levels of essential minerals in rabbits

The effect of tin and lead on levels of essential metals (Zn, Cu, Ca, Fe) in rabbit tissues was compared in relation to the route of administration. Animals received intraperitoneally, or per os, SnCl2 (2 mg Sn/kg) or Pb(CH3COO)2 (3.5 mg Pb/kg) every day for 5 d or for 1 mo. Copper, zinc, iron, and calcium were determined by AAS in the liver, kidneys, spleen, brain, bone marrow, and blood; lead and tin concentration were measured in the blood of animals. Tin and lead administered per os caused either no changes or the decreased concentration of endogenous metals in several tissues. The other route of administration (ip) of both metals generally contributed to the increased storage of essential elements. Blood tin levels of tin treated animals were only about less than or equal to 1/10 of blood lead concentrations of rabbits exposed to lead.     

Unusual evolutionary conservation of 5S rRNA pseudogenes inAspergillus nidulans: Similarity of the DNA sequence associated with the pseudogenes with the mouse immunoglobulin switch region

Summary AllAspergillus nidulans 5S rRNA pseudogenes known so far are the result of integration of an approx. 0.2-kbp-long DNA sequence into the 5S rRNA genes. This sequence, called block C, is present in at least five copies in theA. nidulans genome and seems to be associated either with 5S rRNA genes or pseudogenes. In contrast to the 78% sequence conservation of the C-block in pseudogenes, the truncated 5 halves of the pseudogenes are very highly conserved (96.9–100%). We postulate that the 5S rRNA pseudogenes are still a subject of concerted evolution. The C-block sequence shows similarity to the switch region of the mouse heavy chain immunoglobulin gene. A characteristic motif GGGTGAG is repeated several times in both sequences; the sequence conservation is 63%.     

The effect of yeast cell immobilization on the proportion of selected by-products of ethanol fermentation

Determination was made of the proportion of selected by-products (acetaldehyde, ethyl acetate, methanol, propanol, isobutanol, 2-methyl-butanol, 3-methyl-butanol) of batch and continuous ethanol fermentation carried out with the use of yeastSaccharomyces cerevisiae, strain 0–11, cells immobilized by adsorption on selected carriers (foamed polystyrene, bone shot, beech wood chips, porous glass) as well as by entrapping in calcium alginate and calcium pectinate gel.     

Biosynthesis and preparation of fiveAlternaria metabolites

Metabolites ofAlternaria alternata were produced on rice as a solid substrate, chosen out of 6 substrates as the most useful. Optimal methods of extraction, purification, and separation of 5 metabolites were elaborated, using liquid — liquid partition, column chromatography, and preparative TLC. Alternariol, alternariol methyl ether, and copper salt of tenuazonic acid were obtained as crystals, altertoxin and altenuene as a film.     

DNA degradation at elevated temperatures after plasmid amplification in amino acid-starved Escherichia coli cells

Summary Amino acid starvation of Escherichia coli relA mutants may be used as a method for efficient plasmid DNA amplification. Here we demonstrate DNA degradation which occurs at elevated temperatures (42–43°C) after plasmid amplification in amino acid-starved bacteria. These results may explain the previously described low efficiency of plasmid DNA amplification at elevated temperatures.     

Genome size in wild Pisum species

Genome size was measured in 75 samples of the wild pea species Pisum abyssinicum, P. elatius, P. fulvum and P. humile by ethidium-bromide (EB) flow cytometry (internal standard: Triticum monococcum) and Feulgen densitometry (internal standard: Pisum sativum Kleine Rheinländerin). Total variation of EB-DNA between samples covered 97.7% to 114.9% of the P. sativum value, and Feulgen DNA values were strongly correlated with EB-DNA values (r=0.9317, P < 0.001). Only P. fulvum was homogeneous in genome size (108.9% of P. sativum). Wide variation was observed between samples in P. abyssinicum (100.9–109.7%), P. elatius (97.7–114.9%) and P. humile (98.3–111.1% of P. sativum). In view of the world-wide genome size constancy in P. sativum, the present data are interpreted to show that the pea taxa with variable genome size are genetically inhomogeneous and that the current classification is not sufficient to describe the biological species groups adequately.