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A quaternary ammonium salt (A), a bisquaternary (FB), and an amine oxide (B) were tested for biodegradability using bacterial strains isolated from a municipal waste-water treatment plant Adsorption of the compounds by the cells was distinguished from actual biodegradation. The bisquaternary hard detergent FB was of lower affinity to the bacterial cells in comparison to soft compound A, where adsorption is accompanied by degradation. A soft surfactant, amine oxide (B) is also degraded without marked adsorption by cells. Adsorption of the compouds by the cells is also lower in mineral biodegradation medius where cell membranes are stabilised by ions.
The studies have been performed to analyze the production of β-glucanase by a recombinant strain of Escherichia coli immobilized in different matrices. Porous sintered glass SIRAN®, Ceramic supporting matrices and Broken Pumice stone as well as SIRAN Raschig-rings were examined for the immobilization of whole bacterial cells. The β-glucanase activity of bacteria immobilized in CeramTec PST 5 (4–5 mm) was very low. CeramTec PST 5 (1.5–2.5 mm) was found to be the best carrier compared to all other matrices regarding glucanase production (630 U/ml) and compared to enzyme activity produced by free cells (500 U/ml). Different doses of matrices were applied (2, 5, 7, 10 g/lask) in the form of “matrix weight”. Using 2 g/flask of CeramTec PST 5 (1.5–2.5 mm) yielded enzyme activity of 630 U/ml). CeramTec gives highest operational stability of β-glucanase by repeated batch fermentation to 5 cycles, and activity reached 660 U/ml. Scanning electron microscopy observations showed a high number of vegetative cells that continued growth inside the matrices, indicating that β-glucanase activity improvement was due to the immobilization of the cells.
This is a short survey of the adaptive mutation processes that arise in non- or slowly- dividing bacterial cells and includes: (i) bacterial models in which adaptive mutations are studied; (ii) the mutagenic lesions from which these mutations derive; (iii) the influence of DNA repair processes on the spectrum of adaptive mutations. It is proposed that in starved cells, likely as during the MFD phenomenon, lesions in tRNA suppressor genes are preferentially repaired and no suppressor tRNAs are formed as a result of adaptive mutations. Perhaps the most provocative proposal is (iv) a hypothesis that the majority of adaptive mutations are selected in a pre-apoptotic state where the cells are either mutated, selected, and survive, or they die.
The aim of this study was to evaluate cholesterol uptake by Lb. acidophilus isolates in artificial gastric and duodenal fluids. Studied Lb. acidophilus isolates showed various abilities to uptake cholesterol from MRS broth and artificial GIT fluids. From artificial duodenal fluid Lb. acidophilus isolates removed morę cholesterol (from 0.6% to 2.9%) than from gastric fluid (from 3.8% to 13.5%). Even if bacteria had no possibility to grow in artificial GIT fluids, the cholesterol uptake by bacteria cells took place. Lb. acidophilus isolates survived worse in simulated gastric or duodenal fluids than in MRS broth. The initial number of bacteria ranged from 6.2 log CFU/mL to 7.1 log CFU/mL, and viability in artificial gastric fluid was 60.6-81.9% of log of the initial number of bacteria. Viability in artificial duodenal fluid was 35.1-87.3%.
Low copy number plasmids cannot rely on the random segregation during bacterial cell division. To be stably maintained in the population they evolved two types of mechanisms (i) partition systems (PAR) that actively separate replicated plasmid molecules to the daughter cells and (ii) toxin-andidote systems (TA) that act after cell division to kill plasmid-less cells. Our knowledge of partition systems has been based mainly on analysis of plasmids from Gram-negative bacteria. Now, numerous partition systems of plasmids from Gram-positive bacteria have also been characterized and make significant contribution to our understanding of these mechanisms.
Halophilic bacterial strains isolated from a man-made salt lake in Romania produce a specific exopolysaccharide. This product is synthesized under both static and stirring conditions, and the yield of this exoploysaccharide depends on the composition of the culture medium. The highest amount of polymer was obtained in the presence of casamino acids and yeast extract, under stirring conditions. This polymer has high thermostability, with a melting point at 207°C. The melting process is associated with the thermal degradation of the compound. This polymer is characterized by maximum absorbtion at 260 nm and fluorescence emission at 530 nm. The FT-IR analysis of the polymer revealed the presence of a saccharide structure and of amine and sulfate groups throughout the sugar backbone.
Electron microscopy is a powerful technique for analysis of DNA replication intermediates. However, isolation of replicating DNA molecules from living cells is tricky and difficult, especially in the case of small DNA molecules (such as bacterial plasmids) whose initiation of replication is not easily synchronized. Here a relatively simple and rapid method for efficient isolation of replicating plasmid molecules from unsynchronized Escherichia coli cultures is described. The efficiency of this procedure is high enough for electron microscopy analysis of plasmid replication intermediates appearing in living cells in normal growth conditions. Under optimal conditions, using standard procedures of isolation of plasmid DNA, it is possible to achieve a content of only as few as 0.02 percent of replication intermediates in a plasmid DNA sample. The described method allowed us to enrich up to 100-fold the fraction of replication intermediates suitable for microscopic analysis among all plasmid molecules.
The collection of 314 staphylococcal strains including Staphylococcus aureus and coagulase-negative staphylococci (CNS) was isolated from skin or nasopharynx of healthy people. It was found that the majority of staphylococci possessed the ability to produce slime intensively or moderately, irrespective of ecological niche-nose, throat or skin. Most of them showed the hydrophilic cell surface. However, among S. aureus skin isolates or CNS throat isolates predominated strains with hydrophobic cell surface. There was a slight correlation between slime production and the nature of cell surface among CNS isolates but not among S. aureus strains. It was found that most of slime-producing CNS strains showed hydrophilic cell surface, while slime-negative isolates usually possessed hydrophobic cell surface. Our data suggest that slime production but not cell surface hydrophobicity can be regarded as an essential colonization factor responsible for staphylococci adherence to skin or mucous membranes of upper respiratory tract. These data also suggest that slime production seems to be a general feature of staphylococci isolated from various niches of healthy people.
The role of plasmids of drug resistance R577 and R785 in the development of susceptibility of Escherichia coli K12 strain W1485 to the bactericidal action of serum was studied. Plasmids R577 and R785 were transfered to cells of strain W1485 by means of conjugation. The susceptibility of cells containing the plasmid to serum was compared to that of cells lacking the plasmid. It was found that plasmids R577 and R758 sensitize bacteria to the action of serum.
Spectrin and spectrin-like proteins were found in almost all animal and protozoan cells. Moreover, several reports indicate their presence also in plants. We found that polypeptides reacting with anti-spectrin antibodies are also present in bacteria such as Escherichia coli and Halobacterium salinarium. In Escherichia coli only polypeptide(s) reacting with antibodies directed against a- subunit could be detected while in Halobacterium salinarium polypeptides reactive with antibodies directed against both subunits (α and β) were found. Our data may suggest a presence of spectrin-related polypeptides in early stages of evolution.
This study compares a number of parameters that are important in the ligation of the polymerase chain reaction-amplified DNA inserts into plasmid vectors and their efficient transformation to bacterial cells. The parameters covered were: T4 polynucleotide kinase treatment followed by either the large fragment of E. coli DNA polymerase or T4 DNA polymerase reactions, the amount of T4 DNA ligase, temperature and duration of ligation, molar ratio of insert to vector as well as the total DNA concentration. The results show that the T4 polynucleotide kinase-treated group without further enzymatic manipulation, at an insert to vector ratio of 3:1 gave the highest recombination efficiency when 10 µg/ml DNA and 20 units T4 DNA ligase were applied for ligation for 12 h at 4°C.
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