Search PubMed⌕ Search

Biomedical subjects

S Knapp

Publications and source records attributed to S Knapp.

At least 127 records · Page 7Linked to original sources
PubMed1986

Large, unstable inserts in the chromosome affect virulence properties of uropathogenic Escherichia coli O6 strain 536.

The hemolytic, uropathogenic Escherichia coli 536 (O6:K15:H31) contains two inserts in its chromosome (insert I and insert II), both of which carried hly genes, were rather unstable, and were deleted spontaneously with a frequency of 10(-3) to 10(-4). These inserts were not found in the chromosome of two nonhemolytic E. coli strains, whereas the chromosomal sequences adjacent to these inserts appeared to be again homologous in the uropathogenic and two other E. coli strains. Insert I was 75 kilobases in size and was flanked at both ends by 16 base pairs (bp) (TTCGACTCCTGTGATC) which were arranged in direct orientation. For insert I it was demonstrated that deletion occurred by recombination between the two 16-bp flanking sequences, since mutants lacking this insert still carried a single copy of the 16-bp sequence in the chromosome. Both inserts contained a functional hemolysin determinant. However, the loss of the inserts not only affected the hemolytic phenotype but led to a considerable reduction in serum resistance and the loss of mannose-resistant hemagglutination, caused by the presence of S-type fimbriae (sfa). It is shown that the Sfa-negative phenotype is due to a block in transcription of the sfa genes. Mutants of strain 536 which lacked both inserts were entirely avirulent when tested in several animal model systems.

Adhesins, Escherichia coli↗
PubMed1985

Analysis of the flanking regions from different haemolysin determinants of Escherichia coli.

The haemolysin (hly) determinant of the plasmid pHly152 contains an IS2 element at 469 bp upstream of the hlyC gene. The sequence at the other (right-hand) end (RS) also shows multiple hybridization with the plasmid pHly152 and the chromosome of some Escherichia coli strains but the nucleotide sequence of this region does not reveal the typical properties of an IS element. Similar arrangements in the regions flanking the hly determinant are also found on various Hly plasmids from uropathogenic E. coli strains. Chromosomal hly determinants lack both flanking sequences (IS2 and RS) in the immediate vicinity of the hly genes. The sequences immediately upstream of the hlyC gene have been determined from several chromosomal hly determinants and compared with the corresponding sequence of the hly determinant of the plasmid pHly152. We show that these sequences, which contain one promoter (left promoter, phlyL) in all hly determinants tested, vary considerably although common sequence elements can still be identified. In contrast, only relatively few nucleotide exchanges have been detected in the adjacent structural hlyC genes. The A + T content of the 200 bp sequence upstream of hlyC is very high (72 mol% A + T) but even the structural hly genes show a considerably higher A + T content (about 60 mol%) than the E. coli chromosome on average (50 mol% A + T) suggesting that the hly determinant may not have originated in E. coli.

Amino Acid Sequence↗
PubMed1985

Cloning and characterization of genes involved in production of mannose-resistant, neuraminidase-susceptible (X) fimbriae from a uropathogenic O6:K15:H31 Escherichia coli strain.

The uropathogenic Escherichia coli strain 536 (O6:K15:H31) exhibits a mannose-resistant hemagglutination phenotype (Mrh) with bovine erythrocytes and delayed Mrh with human and guinea pig erythrocytes. Neuraminidase treatment of the erythrocytes abolishes mannose resistant hemagglutination, which is typical for X fimbriae. E. coli strain 536 synthesizes two different fimbriae (Fim phenotype) protein subunits, 16.5 and 22 kilodaltons in size. In addition the strain shows mannose-sensitive hemagglutination and common type I (F1) fimbriae. The cosmid clone E. coli K-12(pANN801) and another nine independently isolated Mrh+ cosmid clones derived from a cosmid gene bank of strain 536 express the 16.5-kilodalton protein band, but not the 22-kilodalton protein, indicating an association of the Mrh+ property with the "16.5-kilodalton fimbriae." All cosmid clones were fimbriated, and they reacted with antiserum produced against Mrh+ fimbriae of the E. coli strain HB101(pANN801) and lacked mannose-sensitive hemagglutination (F1) fimbriae. From the Mrh fim cosmid DNA pANN801, several subclones coding for hemagglutination and X fimbriae were constructed. Subclones that express both hemagglutination and fimbriae and subclones that only code for the hemagglutination antigen were isolated; subclones that only produce fimbriae were not detected. By transposon Tn5 mutagenesis we demonstrated that about 6.5 kilobases of DNA is required for the Mrh+ Fim+ phenotype, and the 1.5- to 2-kilobase DNA region coding for the structural protein of the fimbriae has been mapped adjacent to the region responsible for the Mrh+ phenotype. Two different regions can thus be distinguished in the adhesion determinant, one coding for hemagglutination and the other coding for fimbria formation. Transformation of plasmid DNA from these subclones into a Mrh- Fim- mutant of E. coli 536 and into a galE (rough) strain of Salmonella typhimurium yielded transformants that expressed both hemagglutination and fimbria production.

Animals↗
PubMed1984

Viscosity of xanthan gum solutions at low shear rates.

The viscosity of xanthan gum solutions in the low shear region was investigated with the aid of a Couette instrument. All solutions were highly pseudoplastic . Solutions containing 0.3-0.5% of the gum exhibited a highly ordered phase at very low shear. Viscosity, the degree of pseudoplasticity , and the value of the transition from soft gel to pseudoplastic behavior were directly related to gum concentration. The effect of the addition of a salt on viscosity depended on the xanthan gum concentration. The viscosity of a 0.3% xanthan gum solution was practically unaffected by the salts. Higher gum concentrations exhibited a viscosity increase when salt was present. Concentrations less than 0.3% exhibited a viscosity decrease in the presence of a salt. All viscosity effects seemed to reach limiting values at approximately 10(-3) to 3.3 X 10(-3) N salt. No major differences were observed between sodium chloride, calcium chloride, and sodium citrate in their influence on xanthan gum viscosity.

Chemistry, Pharmaceutical↗
PubMed1984

Multiple copies of hemolysin genes and associated sequences in the chromosomes of uropathogenic Escherichia coli strains.

The O6 serogroup Escherichia coli strain 536 carries two hemolysin (hly) determinants integrated into the chromosome. The two hly determinants are not completely identical, either functionally or structurally, as demonstrated by spontaneous deletion mutants carrying only one of them and by cloning each of the two determinants separately into cosmid vectors. Each hly determinant is independently deleted at a frequency of 10(-4), leading to variants which exhibit similar levels of internal hemolysin but different amounts of secreted hemolysin. The two hly determinants were also identified in the O4 E. coli strain 519. The three E. coli strains 251, 764, and 768, which belong to the serogroup O18, and the O4 strain 367 harbor a single chromosomal hly determinant, as demonstrated by hybridization with hly-gene-specific probes. However, a hybridization probe derived from a sequence adjacent to the hlyC-proximal end of the plasmid pHly 152-encoded hly determinant hybridizes with several additional chromosomal bands in hemolytic O18 and O6 E. coli strains and even in E. coli K-12. The size of the probe causing the multiple hybridization suggests a 1,500- to 1,800-base pair sequence directly flanking hlyC. Spontaneous hemolysin-negative mutants were isolated from strains 764 and 768, which had lost the entire hly determinant but retained all copies of the hlyC-associated sequence.2+.

Base Sequence↗
PubMed1983

Scattering kinetics in a complex tryptophan hydroxylase preparation from rat brainstem raphe nuclei: statistical evidence that the lithium-induced sigmoid velocity function reflects two states of available catalytic potential.

Multiple measures on statistical patterns from a kinetic scattering paradigm for the activities of rat brainstem medial and dorsal raphé nuclear tryptophan hydroxylase preparations are consistent with the previous hypothesis (J. Neural Transmission 45: 1-15, 1979) that the lithium ion induces a state of kinetic bistability in the system. In this context, the lithium-induced sigmoid tetrahydrobiopterin-reaction velocity function is interpreted as a jump between two stable states of differing catalytic potential with a less kinetically accessible domain between them. These dynamics are qualitatively portrayed in three dimensions by Thom's classical cusp catastrophe.

Animals↗
PubMed1983

Lithium and chlorimipramine differentially alter the stability properties of tryptophan hydroxylase as seen in allosteric and scattering kinetics.

Compared to control, both lithium and chlorimipramine (CMI) slow the frequency of variation in tryptophan hydroxylase (TPOH) velocity functions measured across increasing (nearly physiological) cofactor concentrations or across continuous (long residence) time. The common effect appears to be generated by different statistical mechanisms in view of the diverse patterns observed when, for multiple 60-point experiments for each condition, mean frequencies were plotted against mean amplitudes of variation: lithium constricted the frequencies over a range of amplitudes; CMI fixed amplitude across the range of frequencies. Initial rate, short residence time experiments suggest that at the micromolecular level, before the emergence of time-dependent macro-kinetic organization as seen in the long residence time assays, lithium may uncouple the elementary constituents of the system, allowing subsequent phase gathering and emergent stability around a statistically dominant frequency, whereas CMI may augment coupling, leading to unstable complexity in the population dynamics. The relationship between these effects and allosteric kinetic functions is discussed.

Allosteric Regulation↗
PubMed1983

Spontaneous deletions and flanking regions of the chromosomally inherited hemolysin determinant of an Escherichia coli O6 strain.

The hemolytic Escherichia coli strain 536 (O6) propagates spontaneous hemolysin-negative mutants at relatively high rates (10(-3) to 10(-4)). One type of mutant (type I) lacks both secreted (external) and periplasmic (internal) hemolysin activity (Hlyex-/Hlyin-) and in addition shows no mannose-resistant hemagglutination (Mrh-), whereas the other type (type II) is Hlyex-/Hlyin+ and Mrh+. The genetic determinants for hemolysin production (hly) and for mannose-resistant hemagglutination (mrh) of this strain are located on the chromosome. Hybridization experiments with DNA probes specific for various parts of the hly determinant reveal that mutants of type I have lost the total hly determinant, whereas those of type II lack only part of the hlyB that is essential for transport of hemolysin across the outer membrane. Using a probe that contains the end sequence of the plasmid pHly152-encoded hly determinant (adjacent to hlyB), we determined that a related sequence flanks also the hlyB-distal end of the chromosomal hly determinant of E. coli 536. In addition several other similar or even identical sequences are found in the vicinity of the hlyC- and the hlyB-distal ends of both the chromosomal and the plasmid hly determinants.

Base Sequence↗
PubMed1981

Strain differences in kinetic and thermal stability of two mouse brain tryptophan hydroxylase activities.

Levels of forebrain serotonin (5-HT), tryptophan, 5-hydroxyindoleacetic acid (5-HIAA) and hydroxylase cofactor (BH4) were comparable in two experimental mouse strains (A/J and C57Bl/6J) despite 2-3-fold differences in vitro in the relative activities of forebrain and midbrain tryptophan-5-monooxygenase (TPOH; EC 1.14.16.4). The enzyme activities did not differ with respect to Km for cofactor at saturating levels, but manifested different degrees of cooperativity with respect to cofactor when examined with BH4 concentrations within a physiological range. They differed also in the frequency and amplitude of kinetic variation around comparable mean velocity slopes across cofactor and time; in resistance to pre-incubation inactivation and responsiveness to its facilitation by calcium; in molecular weight heterogeneity as reflected in the distribution of molecular weight forms by gel diffusion chromatography; and in the number of peaks in power spectral analysis of kinetic variation patterns. Although the potential roles of small-molecule ligand and/or regulator proteins have not been ruled out, we hypothesize that differences in conformational stability underlie the differences in regulatory properties and make one enzyme activity more vulnerable to occlusive influences in vivo.

Animals↗
PubMed1981

Mouse midbrain tryptophan hydroxylase: strain differences in variational properties.

Biochemical studies of kinetic conformations and assessment of statistical patterns of variation around mean velocity slopes were used to characterize midbrain tryptophan hydroxylase (TPOH) from two behaviorally different mouse strains, C57B1/6J and A/J. The data suggest that the two strains manifest forms of TPOH with different stabilities and regulatory properties.

Animals↗