Ribosomal and non-ribosomal resistance to oxazolidinones: species-specific idiosyncrasy of ribosomal alterations

A derivative of Mycobacterium smegmatis, which carries only one functional rRNA (rrn) operon, was used to isolate mutants resistant to the ribosome-targeted antibiotic linezolid. Isolation and characterization of linezolid-resistant clones revealed two classes of mutants. Ribosomes from class I mutants are resistant to oxazolidinones in an in vitro peptidyl transferase assay, indicating that resistance maps to the ribosome component. In contrast, ribosomes from class II mutants show wild-type susceptibility to a linezolid derivative in vitro, pointing to a non-ribosomal mechanism of resistance. Introduction of a wild-type ribosomal RNA operon into linezolid-resistant strains restored linezolid sensitivity in class I mutants, indicating that resistance (i) maps to the rRNA and (ii) is recessive. Sequencing of the entire rrn operon identified a single nucleotide alteration in 23S rRNA of class I mutant strains, 2447G --> T (Escherichia coli numbering). Introduction of mutant rrl2447T into M. smegmatis rrn- resulted in a linezolid-resistant phenotype, demonstrating a cause-effect relationship of the 2447G --> T alteration. The 2447G --> T mutation, which renders M. smegmatis linezolid resistant, confers lethality in E. coli. This finding is strong evidence of structural and pos-sibly functional differences between the ribosomes of Gram-positive and Gram-negative bacteria. In agreement with the results of the in vitro assay, class II mutants show a wild-type sequence of the complete rRNA operon. The lack of cross-resistance of the class II mutants to other antibiotics suggests a resistance mechanism other than activation of a broad-spectrum multidrug transporter.     

Evidence for an early prokaryotic origin of histones H2A and H4 prior to the emergence of eukaryotes.

Histones have been identified recently in many prokaryotes. These histones, unlike their eukaryotic homologs, are of a single uniform type that is thought to resemble the archetypal ancestor of the eukaryotic histone family. In this paper we report the finding, the cloning and the phylogenetic analysis of the sequence of a prokaryotic histone from the hyperthermophile Methanopyrus kandleri . Unlike previously described prokaryotic histones, the Methanopyrus sequence has a novel structure consisting of two tandemly repeated histone fold motifs in a single polypeptide. Sequence analyses indicate that the N-terminal repeat is most closely related to eukaryotic H2A and H4 histones, whereas the C-terminal repeat resembles that found in prokaryotic histones. These results imply an early divergence within the histone gene family prior to the emergence of eukaryotes and may represent an evolutionary step leading to eukaryotic histones.     

The ultrastructure of Blastocystis galli from chickens

The ultrastructure stages of Blastocystis galli were studied in chicken's intestine and in laboratory cultures. There were found morphological structures: surface coat (cell from chickens' intestine showed a very thick surface coat); cell membrane--there were some small electron-opaque deepening "pockets" on the membrane; inner membrane; endoplasmic reticulum with attached ribosomes, which present in the cytoplasm; all cells contained numerous of small vacuoles and large glycogen inclusions in cytoplasm; mitochondria with tubular cristae; nucleus with granules condensed chromatin; central vacuole; Golgi complex was represented by number of plates grouped in a pite; the cyst-like forms were surrounded by multilayered wall.     

Isoform-specific O-glycosylation by murine UDP-GalNAc:polypeptide N- acetylgalactosaminyltransferase-T3, in vivo

Multiple isoforms of UDP-GalNAc:polypeptide N-acetylgalactosaminyl- transferase (ppGaNTase) have been cloned and expressed from a variety of organisms. In general, these isoforms display different patterns of tissue-specific expression, but exhibit overlapping substrate specificities, in vitro . A peptide substrate, derived from the sequence of the V3 loop of the HIV gp120 protein (HIV peptide), has previously been shown to be glycosylated in vitro exclusively by the ppGaNTase-T3 (Bennett et al. , 1996). To determine if this isoform- specificity is maintained in vivo , we have examined the glycosylation of this substrate when it is expressed as a reporter peptide (rHIV) in a cell background (COS7 cells) which lacks detectable levels of the ppGaNTase-T3. Glycosylation of rHIV was greatly increased by coexpression of a recombinant ppGaNTase-T3. Overexpression of ppGaNTase- T1 yielded only partial glycosylation of the reporter. We have also determined that the introduction of a proline residue at the +3 position flanking the potential glycosylation site eliminated ppGaNTase- T3 selectivity toward rHIV observed both in vivo and in vitro .      

A new Spanish species of Linaria Miller (Scrophulariaceae)

Linaria arabiniana sp. nov. is described from coastal sand dunes of a very reduced area in Alicante Province (south-eastern part of the Iberian Peninsula). It is a perennial herb characterized by its 3–4-verticillate leaves, glabrous stems, large violet or rarely yellow flowers, and bicoloured usually flat seeds. Morphological characteristics and taxonomic affinities of this taxon are discussed, as are data on its ecology, phytosociology, and eventual conservation features.     

Transformation and transduction in Bacillus subtilis strains with the BsuR restriction-modification system by modified and unmodified pUB110 plasmid DNA

Plasmid pUB110 DNA is restricted and modified during transformation of Bacillus subtilis cells possessing the BsuR system of restriction-modification. Restriction has comparatively little influence on the frequency of plasmid transformation only reducing it 20 times. The frequency of transduction of nonmodified plasmid DNA into modifying recipient cells, using phage Ar9 is also reduced a little. The frequency of transduction of chromosomal markers is much more lowered under the same experimental conditions.     

Bulbing in Long-day Onion (Allium cepa L.) Cultured In Vitro: Comparison Between Sugar Feeding and Light Induction

Bulbs were obtained on onion plants cultured in vitro. No bulbinghappened under long days with fluorescent light and 30–40g l^–1 sucrose. Bulb formation was mainly dependent eitheron sucrose concentration in the culture medium, or on lightspectral composition. Raising the sucrose concentration from40 to 120 g l^–1 increased plant basal swelling and stoppedfurther vegetative development. These plants were not dormant.When fluorescent light was enriched in incandescence duringa long day period, bulbs were obtained in two months and underwenta consecutive dormancy. Bulb, dormancy, light spectral quality, photoperiod, R: FR ratio, sugar, tissue culture     

A new species of Blastocystis anseri (Protista: Rhizopoda) from domestic geese]

A new species, Blastocystis anseri, was found in domestic goose. Sizes of blastocyst in culture are 7.5-46.2 x 7.5-46.2 m. Method of cultivation of Blastocystis anseri on biphase egg medium was worked out. Liquid phase can be made of Hank's solution or 199 medium with an addition of 30-40% hen or bovine serum. Optimum temperature for cultivation is 39 +/- 0.5 degree, ph 7.0-7.2.