Phase variation of type 1 pili (fimbriae) was studied during the in vivo growth of Escherichia coli in two animal models. In the first, a heavily piliated urinary tract isolate (strain 149) was placed in 1-cm polypropylene chambers sealed with 0.22-,um-pore-size filters. The chambers were surgically implanted intraperitoneally in mice and recovered at various times. Piliation, as determined by electron microscopy and by measuring the minimum number of bacteria needed to produce mannose-sensitive hemagglutination, gradually decreased, and by day 5, most of the organisms were nonpiliated. In the second model, piliated and nonpiliated E. coli phase variants were inoculated into the bladders of BALB/c mice via urinary catheters, and their fate in the lower urinary tract was studied. Viable counts of bladder homogenates revealed that piliated phase variants were significantly more effective in colonizing the bladder urothelium than were their nonpiliated counterparts. Specific antibody to type 1 pili prevented colonization by the piliated organisms. After inoculation of piliated variants, the bladder-associated bacteria gave rise to approximately 80% mannose-sensitive hemagglutinationpositive colonies, and immunocytochemistry of bladder lavages revealed large numbers of type 1 piliated bacteria adhering to the bladder transitional cells. Electron microscopy confirmed the presence of piliated bacteria in association with the bladder urothelium. The urine of these mice, whose bladders were colonized with piliated bacteria, frequently showed no growth, and when bacteria were present, strain 149 yielded less than 30% hemagglutination-positive colonies. The results suggest that for some E. coli strains, phase variation may be a factor in determining the fate of the E. coli in the urinary tract and that the urine may not necessarily reflect the bacteriologic state of the bladder mucosa.

Abstract. Ten years of ERA5 reanalysis data are combined with met-mast and lidar observations from 10 offshore platforms to investigate low-level jet characteristics over the Dutch North Sea. The objective of this study is to combine the best of two worlds: (1) ERA5 data with a large spatiotemporal extent but inherent accuracy limitations due to a relatively coarse grid and an incomplete representation of physical processes and (2) observations that provide more reliable estimates of the measured quantity but are limited in both space and time. We demonstrate the effect of time and range limitations on the reconstructed wind climate, with special attention paid to the impact on low-level jets. For both measurement and model data, the representation of wind speed is biased. The limited temporal extent of observations leads to a wind speed bias on the order of ±1 m s−1 as compared to the long-term mean. In part due to data-assimilation strategies that cause abrupt discontinuities in the diurnal cycle, ERA5 also exhibits a wind speed bias of approximately 0.5 m s−1. The representation of low-level jets in ERA5 is poor in terms of a one-to-one correspondence, and the jets appear vertically displaced (“smeared out”). However, climatological characteristics such as the shape of the seasonal cycle and the affinity with certain circulation patterns are represented quite well, albeit with different magnitudes. We therefore experiment with various methods to adjust the modelled low-level jet rate to the observations or, vice versa, to correct for the erratic nature of the short observation periods using long-term ERA5 information. While quantitative uncertainty is still quite large, the presented results provide valuable insight into North Sea low-level jet characteristics. These jets occur predominantly for circulation types with an easterly component, with a clear peak in spring, and are concentrated along the coasts at heights between 50 and 200 m. Further, it is demonstrated that these characteristics can be used as predictors to infer the observed low-level jet rate from ERA5 data with reasonable accuracy.

Type 1 pili in Escherichia coli undergo phase variation in which individual cells in a population reversibly switch between piliated (Pil+) and nonpiliated (Pil-) states. The switching process is mediated by an invertible DNA fragment which contains the promoter for fimA, the gene encoding the major structural subunit of type 1 pili. Although type 1 pili randomly phase vary in broth cultures, many clinical isolates of E. coli do not express type 1 pili when cultured on agar media. We investigated the role of the invertible element and the upstream genes, fimB and fimE, in the agar-mediated suppression of pili in an agar-negative clinical isolate, strain 149. Southern hybridization and polymerase chain reaction analyses of the fimA promoter region in broth-grown 149 cells indicated that the invertible element was present in orientations corresponding to both Pil+ and Pil- phenotypes. In contrast, only one orientation of the invertible element, corresponding to the Pil- phenotype, was observed in strain 149 cells cultured on agar. A second clinical isolate, strain 2-7, which expresses type 1 pili on agar was also examined; the invertible element was found in both the Pil+ and Pil- orientations during growth of this strain on agar as well as in broth. The introduction of the fim gene cluster from strain J96 on a multicopy plasmid into agar-negative strain 149 resulted in the production of both J96 and 149 pili during growth on agar. Experiments with subclones of the J96 genes indicated that the presence of an intact fimB gene allowed strain 149 pili to be produced on agar. Differences in pilus production between agar and broth cultures appear to be the result of differential transcription of fimB and fimE under the two growth conditions. In contrast, the pattern of expression of these genes in agar phase-variable strain 2-7 did not differ between broth- and agar-grown cells.

The piliation and hemagglutination properties of 54 consecutive Escherichia coli isolates from women with recurrent urinary tract infections were studied. Mannose-sensitive hemagglutination (MSHA) of guinea pig erythrocytes, characteristic of type 1-piliated bacteria, was produced by 75% of the isolates, 32% produced mannose-insensitive hemagglutination, and 14% produced no hemagglutination reaction. The production of * Corresponding author.

The effect of the bacterial cytolytic toxin, streptolysin O (SLO), on rabbit erythrocyte membranes, liposomes, and lipid dispersions was examined. SLO produced no gross alterations in the major erythrocyte membrane proteins or lipids. However, when erythrocytes were treated with SLO and examined by electron microscopy, rings and "C"-shaped structures were observed in the cell membrane. The rings had an electron-dense center, 24 nm in diameter, and the overall diameter of the structure was 38 nm. Ring formation also occurred when erythrocyte membranes were fixed with glutaraldehyde and OsO4 before the addition of toxin. In contrast, rings were not seen when erythrocytes were treated with toxin at 0~indicating that adsorption of SLO to the membrane is not sufficient for ring formation since toxin is known to bind to erythrocytes at that temperature. The ring structures were present on lecithin-cholesterol-dicetylphosphate liposomes after SLO treatment, but there was no release of the trapped, internal markers, K2CrO4 or glucose. The crucial role of cholesterol in the formation of rings and C's was demonstrated by the fact that these structures were present in toxin-treated cholesterol dispersions, but not in lecithin-dicetylphosphate dispersions nor in the SLO preparations alone. The importance of cholesterol was also shown by the finding that no rings were present in membranes or cholesterol dispersions which had been treated with digitonin before SLO was added. Although the rings do not appear to be "holes" in the membrane, a model is proposed which suggests that cholesterol molecules are sequestered during ring and C-structure formation, and that this process plays a role in SLO-induced hemolysis.Streptolysin O (SLO) is a bacterial toxin produced by virtually all strains of Streptococcus pyogenes. The toxin is a protein with a molecular weight of approximately 60,000 daltons, and is characteristic of a group of cytolytic toxins known as the oxygen-labile toxins (3). The toxins in this group are produced by several different gram-positive bacteria, and possess a number of common properties: they are activated by SH compounds: they appear to be antigenically related; and their biological activity is completely inhibited by low concentrations of cholesterol and certain related sterols.Hemolysis occurs within minutes after the addi-160 THE JOURNAL OF CELL BIOLOGY 9 VOLUME 67, 1975 9 pages 160-173 tion of SLO to erythrocytes, and toxic effects of SLO on several types of mammalian cells in culture have been demonstrated (13). The speed with which sensitive cells are affected by the toxin suggests that the cell membrane is the primary site of action. Several lines of indirect evidence suggest that membrane cholesterol is the binding site and/or target of SLO action. Only those cells which contain cholesterol in their membranes are susceptible to the toxin, SLO is "inactivated" only by the membrane lipid fraction which contains cholesterol, and the addition of exogenous cholesterol to SLO inhibits toxin action....