Structural and immunochemical studies on the lipopolysaccharide of the ‘T-antigen’-containing mutant Proteus mirabilis R14/1959

Abstract In DOC-PAGE, lipopolysaccharide (LPS) of Proteus mirabilis R14/1959 (Rb-type) mutant showed a ladder-like migration pattern indicating the presence of a high molecular weight polysaccharide chain. The isolated polysaccharide, called T-antigen because of similarity with the T1 chain of Salmonella friedenau LPS, contained d -glucose, d -galacturonic acid ( d -GalA), and d -GlcNAc in molar ratios 2:1:1 and was structurally different from the O-antigen of the parental S-strain P. mirabilis S1959 but identical to the O-antigen of another S-strain Proteus penneri 42. The importance of a d -GalA( l -Lys)-containing epitope, most likely present in the core region of LPS, and of GalA present in the T-antigen chain in manifesting the serological specificity of P. mirabilis R14/1959 were revealed using rabbit polyclonal homologous and heterologous R- and O-specific antisera and the appropriate antigens, including synthetic antigens which represent partial structures of various Proteus LPS.     

SEM observations on polar body elimination and changes of the oocyte surface during egg maturation in the newt Triturus alpestris (Amphibia,Urodela)

Summary We have studied the surface of the animal half of ovulated newt eggs recovered from different portions of the oviduct. The germinative area, about 40 m diameter, is localized in the region of the whitish polar spot, about 450 m diameter. The structural changes in the germinative area are connected with the formation and extrusion of the first and second polar bodies. Of the two types of oviductal eggs observed, those covered with microvilli (type 1) were found only in the ostial portions of the oviduct, whilst those covered with microfolds (type 11) were found more distally. The structural difference between these two types may be related to the known reduction in surface area of the cell membrane during oocyte maturation. Offprint requests to: W. Kilarski     

The R gene product of bacteriophage lambda is the murein transglycosylase

Summary The radioactively labeled proteins synthesised in Escherichia coli minicells infected by bacteriophage R and R ⁺ were compared by polyacrylamide gel electrophoresis. R mutants, which have lost the ability to lyse host cells, lack a polypeptide of molecular weight 17.5 kD corresponding to the molecular weight of murein transglycosylase — a bacteriolytic enzyme from lysates which we have described previously. It has been shown by direct comparison using radio-labeled enzyme that transglycosylase comigrates with the R gene product. The enzyme was endetectable in induced cultures of E. coli W3350 su ^o (cI857 Ram5) and C600 (cI857 acR301), while it was present in a R _– ⁺ mutant lysate. We conclude that the transglycosylase is the R gene product.Abbreviations Muropeptide CA GlcNac-1-4-1,6-anhydro-MurNac-L-Ala-D-Glu-msA₂pm-D-Ala - muropeptide CB GlcNac-MurNac-GlcNac-1,6-anhydro-MurNac in which the carboxyl groups of MurNac and 1,6-anhydro-MurNac are substituted by the tetrapeptide L-Ala-D-Glu-msA₂pm-D-Ala - muropeptide C3 dimer of the two units GlcNac-MurNac-L-Ala-D-Glu-msA₂pm-D-Ala which are connected by D-D peptide bond between D-Ala and msA₂pm - GlcNac N-acetyl-D-glucosamine - MurNac N-acetylmuramic acid - msA₂pm meso-diaminopimelic acid - rivanol 6,9-diamino-2-ethoxyacridine lactate - SDS sodium dodecyl sulfate     

Effect of DNA gyrase inhibitors on gene expression of the cysteine regulon

Summary Nalidixic acid inhibits the expression of those cysteine genes which are regulated by the cysB product, it has no affect, however, on the constitutively expressed cysE gene. The expression of cysteine genes in a strain carrying a mutation in the nalA locus is resistant to this drug. Novobiocin affects the expression of cysteine genes similarly to nalidixic acid. The effect of nalidixic acid on the expression of genes in a cysteine constitutive mutant was studied.     

RAPD analysis of the interspecific somatic hybrids: Solanum bulbocastanum (+) S. tuberosum

The diploid Mexican species S. bulbocastanum (blb) was used as a source of late blight resistance in somatic hybridization with the potato (S. tuberosum, tbr) dihaploid H-8105. The produced 2x blb (+) 2x tbr H-8105 somatic hybrids did not retain the blb parent's characteristic high resistance to P. infestans. The revealed aneuploidy of blb (+) tbr H-8105 hybrids indicated a possible loss of individual blb chromosome(s) carrying the resistance genes. Four hybrid clones differing in terms of their ploidy, morphology and growth potential were analysed for the presence of all twelve blb chromosomes (linkage groups). The RAPD markers assigned to particular chromosomes were selected on the basis of the linkage map of S. bulbocastanum constructed by Naess et al., Mol. Gen. Genom. 265 (2001) 694-704. Of the 86 markers analysed, twelve (14%) were common for blb and H-8105, while 34 (40%) and 40 (46%) markers were specific for the blb and H-8105 genome, respectively; this confirms the differences between the nuclear genomes of the two species. Seventeen markers (20%) present in one or the other parent were absent in the hybrids, and only one new marker was found in the hybrids. The poorly growing, aneuploid and chimeric hybrids had the same band profiles as the well growing, morphologically normal hybrids, except for two bands that were present only in normal hybrids. The presence of eleven blb linkage groups in the blb (+) tbr H-8105 hybrids was confirmed. The markers specific for the second linkage group (chromosome 2) of blb were not present in the RAPD patterns of the somatic hybrids analysed, suggesting a loss or rearrangement of this chromosome in the combined nuclear genome of the hybrids.     

Identification of the Escherichia coli sohB gene, a multicopy suppressor of the HtrA (DegP) null phenotype.

We cloned and sequenced the sohB gene of Escherichia coli. The temperature-sensitive phenotype of bacteria that carry a Tn10 insertion in the htrA (degP) gene is relieved when the sohB gene is present in the cell on a multicopy plasmid (30 to 50 copies per cell). The htrA gene encodes a periplasmic protease required for bacterial viability only at high temperature, i.e., above 39 degrees C. The sohB gene maps to 28 min on the E. coli chromosome, precisely between the topA and btuR genes. The gene encodes a 39,000-Mr precursor protein which is processed to a 37,000-Mr mature form. Sequencing of a DNA fragment containing the gene revealed an open reading frame which could encode a protein of Mr 39,474 with a predicted signal sequence cleavage site between amino acids 22 and 23. Cleavage at this site would reduce the size of the processed protein to 37,474 Mr. The predicted protein encoded by the open reading frame has homology with the inner membrane enzyme protease IV of E. coli, which digests cleaved signal peptides. Therefore, it is possible that the sohB gene encodes a previously undiscovered periplasmic protease in E. coli that, when overexpressed, can partially compensate for the missing HtrA protein function.     

Occurrence of airborne Cladosporium and Alternaria spores in Southern and Central Poland in 1995–1996

The concentration of airborne spores of Cladosporium and Alternaria has been investigated at five monitoring stations situated in cities from the foot of the Tatra Mountains to central Poland along a south-north transect (Zakopane, Kraków, Ostrowiec witokrzyski, Warszawa, Pozna) i.e. from a height of 900 m to 80 m above sea level. The aerobiological monitoring of fungal spores was performed by means of five Burkard volumetric spore traps.Cladosporium spores were dominant at all the stations. The highest Cladosporium and Alternaria spore concentrations were observed at all the sites in July and August, except at Warszawa in both years and at Pozna in 1995 where the maximum of Cladosporium spores occurred in June and July, and at Ostrowiec witokrzyski in 1995 where the maximum was found in July, August and September.Fungal spore concentrations in Zakopane and Kraków were significantly lower than those in Ostrowiec witokrzyski, Warszawa and Pozna and periods of abundant Cladosporium spore occurrence were different in these two groups of monitoring stations.     

The relationship between airborne pollen and fungal spore concentrations and seasonal pollen allergy symptoms in Cracow in 1997–1999

The investigation of airborne pollen and fungalspore concentrations was carried out in Cracowbetween 1997–1999. For this study thevolumetric method has been employed (Burkard).At the same time the clinical diagnosis ofpollen allergy in 40 patients was obtained onthe basis of an interview, positive skin pricktests with pollen extracts and increasedspecific IgE level. An increase in seasonalallergy symptoms in all patients occurred fromthe middle of May to the middle of August.Eighty eight percent of the patients (35 out of40) were sensitive to Poaceae pollen and about50% of them were additionally sensitive totree and herb pollen excluding grasses. Forpatients with additional allergy to tree pollenthe seasonal symptoms started at the end ofMarch (Betula) while for patients withadditional allergy to herb pollen it wasextended to the middle of September (Artemisia).Five people out of 40 revealed positive skinreactions to Alternaria spores and anincrease in specific IgE level. Positive skinreaction to Cladosporium spores with noincrease in specific IgE level occurred in 2patients. The increase in seasonal allergysymptoms in people sensitive to Alternariaspores noted in July and August could becaused not only by these spores but also byPoaceae pollen.