Receptor binding studies disclose a novel class of high-affinity inhibitors of the Escherichia coli FimH adhesin

Mannose-binding type 1 pili are important virulence factors for the establishment of Escherichia coli urinary tract infections (UTIs). These infections are initiated by adhesion of uropathogenic E. coli to uroplakin receptors in the uroepithelium via the FimH adhesin located at the tips of type 1 pili. Blocking of bacterial adhesion is able to prevent infection. Here, we provide for the first time binding data of the molecular events underlying type 1 fimbrial adherence, by crystallographic analyses of the FimH receptor binding domains from a uropathogenic and a K-12 strain, and affinity measurements with mannose, common mono- and disaccharides, and a series of alkyl and aryl mannosides. Our results illustrate that the lectin domain of the FimH adhesin is a stable and functional entity and that an exogenous butyl alpha- D-mannoside, bound in the crystal structures, exhibits a significantly better affinity for FimH (K-d = 0.15 muM) than mannose (K-d = 2.3 muM). Exploration of the binding affinities of alpha-D-mannosides with longer alkyl tails revealed affinities up to 5 nM. Aryl mannosides and fructose can also bind with high affinities to the FimH lectin domain, with a 100-fold improvement and 15-fold reduction in affinity, respectively, compared with mannose. Taken together, these relative FimH affinities correlate exceptionally well with the relative concentrations of the same glycans needed for the inhibition of adherence of type 1 piliated E. coli. We foresee that our findings will spark new ideas and initiatives for the development of UTI vaccines and anti-adhesive drugs to prevent anticipated and recurrent UTIs.

Uptake and metabolism of ginsenoside Rh2 and its aglycon protopanaxadiol by Caco-2 cells

The uptake and metabolism profiles of ginsenoside Rh2 and its aglycon protopanaxadiol (ppd) were studied in the human epithelial Caco-2 cell line. High-performance liquid chromatography-mass spectrometry was applied to determine Rh2 and its aglycon ppd concentration in the cells at different pH, temperature, concentration levels and in the presence or absence of inhibitors. Rh2 uptake was time and concentration dependent, and its uptake rates were reduced by metabolic inhibitors and influenced by low temperature, thus indicating that the absorption process was energy-dependent. Drug uptake was maximal when the extracellular pH was 7.0 for Rh2 and 8.0 for ppd. Rh2 kinetic analysis showed that a non-saturable component (K-d 0.17 nmol (.) h(-1) (.) mg(-1) protein) and an active transport system with a K-m of 3.95 mumol (.) l(-1) and a V-max of 4.78 nmol(.)h(-1) (.)mg(-1) protein were responsible for the drug uptake. Kinetic analysis of ppd showed a non-saturable component (K-d 0.78 nmol (.) h(-1) (.) mg(-1) protein). It was suggested that active extrusion of P-glycoprotein and drug degradation in the intestine may influence Rh2 bioavailability.

Prostanoids as friends, not foes: Further evidence from the interference by cycloxygenase-inhibitory drugs when inducing tolerance to experimental arthritigens in rats

Pharmacologists have generally been prejudiced against prostanoids, uncritically accepting their suppression as desirable therapy, especially for ‘quick-fix’ analgesia. This myopic perception for a long time ignored (a) the essentiality of prostanoid precursors in nutrition, (b) the physiological protective functions of natural prostaglandins (PGs) (vasculature, stomach, kidney), (c) resolution of inflammation after the expression of COX-2 and (d) increasing therapeutic use of either synthetic PGs (for erectile dysfunction, opthalmic disorders, inducing parturition, etc) or their natural precursors, e.g., ω3-rich polyunsaturated oils, to treat arthritis. Experimental studies in rats have indicated that prostaglandins (E series) are (i) useful, perhaps auto-regulators of established immunoreactivity and (ii) able to amplify (or even induce) anti-inflammatory activity with other agents. Furthermore, anti-prostanoid therapy (APT) can be arthritigenic!!, interfering with the acquisition of tolerance to some arthritigens. For patients with rheumatoid arthritis this additional side-effect of APT, barely recognised to date, may actually perpetuate their arthritis by impairing prostanoid-mediated remission processes. Hopefully, recent adverse publicity about COX-2 inhibitory drugs might stimulate serious re-assessment of some traditional anti-inflammatory therapies with low APT activity for the management of both acute pain (non-addictive cannabinoids, celery seed, etc.) and chronic inflammation, e.g., Lyprinol® (a mussel lipid extract).

Late pleistocene to holocene composite speleothem O-18 and C-13 chronologies from south island, new Zealand-did a global younger dryas really exist?

Oxygen and carbon data from eight stalagmites from northwest South Island are combined to produce composite records of delta(18)O and delta(13)C from 23.4 ka to the present. The chronology is anchored by 43 thermal ionization mass spectrometry (TIMS) uranium series ages. Delta O-18 values are interpreted as having a first order positive relationship to temperature, but also to be influenced by precipitation in a complex manner. Delta C-13 is interpreted as responding negatively to increases in atmospheric CO, concentration, biological activity and precipitation amount. Six climatic phases are recognized. After adjustment of 1.2parts per thousand for the ice volume effect, the delta(18)O record between 23 and 18 ka varies around -3.72parts per thousand compared to the Holocene average of -3.17parts per thousand. Late-glacial warming commenced between 18.2 and 17.8 ka and accelerated after 16.7 ka, culminating in a positive excursion between 14.70 and 13.53 ka. This was followed by a significant negative excursion between 13.53 and 11.14 ka of up to 0.55parts per thousand depth that overlapped the Antarctic Cold Reversal (ACR) and spanned the Younger Dryas (YD). Positive delta(18)O excursions at 11.14 ka and 6.91-6.47 ka represent the warmest parts of the Holocene. The mid-Holocene from 6 to 2 ka was marked by negative excursions that coincide with increased glacial activity in the South Island. A short positive excursion from 0.71 to 0.57 ka was slightly later than the Medieval Warm Period of Europe. Delta C-13 values were high until 17.79 ka after which there was an abrupt decrease to 17.19 ka followed by a steady decline to a minimum at 10.97 ka. Then followed a general increase, suggesting a drying trend, to 3.23 ka followed by a further general decline. The abrupt decrease in delta-values after 17.79 ka probably corresponds to an increase in atmospheric CO2 concentration, biological activity and wetness at the end of the Last Glaciation, but the reversal identified in the delta(18)O record from 13.53 to 11.14 ka was not reflected in delta(13)C changes. The lowest delta(13)C values coincided with the early Holocene climatic suboptimum when conditions were relatively wet as well as mild. Major trends in the delta(18)O(c) record are similar to the Northern Hemisphere, but second order detail is often distinctly different. Consequently, at the millennial scale, a more convincing case can be made for asymmetric climatic response between the two hemispheres rather than synchronicity. (C) 2004 Elsevier B.V. All rights reserved.

A new estimate for the composition of weathered young upper continental crust from alluvial sediments, Queensland, Australia

We present new major element, trace element and Nd-isotope data for 30 alluvial sediments collected from 25 rivers in Queensland, E Australia. Samples were chosen to represent drainage from the region's most important lithologies, including Tertiary intraplate volcanic rocks, a Cretaceous igneous province (and sedimentary rocks derived thereof) as well as Proterozoic blocks. In most chemical and isotopic aspects, the alluvial sediments represent binary or ternary mixing relationships, with absolute abundances implied to reflect the proportion of lithologies in the catchments. When averaged, the studied sediments differ from other proxies of upper continental crust (UCC) mainly in their relative middle rare earth element enrichment (including an elevated Sm/Nd ratio), higher relative Eu abundance and higher Nb/Ta ratio. These features are inherited from eroded Tertiary intraplate basalts, which commonly form topographic highs in the studied region. Despite the high degree of weathering strong to excellent coherence between similarly incompatible elements is found for all samples. From this coherence, we suggest revisions of the following upper crustal element ratios: Y/Ho = 26.2, Yb/Tm = 6.37, Th/W = 7.14, Th/Tl = 24 and Zr/Hf = 36.9. Lithium, Rb, Cs and Be contents do not seem depleted relative to UCC, which may reflect paucity of K-feldspar in the eroded catchments. Nickel, Cr, Pb, Cu and Zn concentrations are elevated in polluted rivers surrounding the state capital. River sediments in the Proterozoic Georgetown Inlier are elevated in Pb, Cu and Zn but this could be a natural phenomenon reflecting abundant sulphide mineralisation in the area. Except for relative Sr concentrations, which broadly anticorrelate with mean annual rainfall in catchments, there is no obvious relationship between the extent of weathering and climate types, which range from and to tropical. The most likely explanation for this observation is that the weathering profiles in many catchments are several Myr old, established during the much wetter Miocene period. The studied sediment compositions (excluding those from the Proterozoic catchments) are used to propose a new trace element normalisation termed MUQ (MUd from Queensland), which serves as an alternative to UCC proxies derived from sedimentary rocks. Copyright (C) 2005 Elsevier Ltd

Origin of the Archean Sask craton and its extent within the Trans-Hudson orogen: evidence from Pb and Nd isotopic compositions of basement rocks and post-orogenic intrusions

U-Pb zircon ages from the exposed Sask craton are 2450-3100 Ma, from the Peter Lake Domain 2575-2640 Ma, and from rocks of the Trans-Hudson orogen 1840-1880 Ma. U-Pb monazite and zircon ages of post-orogenic pegmatites and aplites are 1770-1800 Ma. Common Pb and Sm-Nd isotopic compositions of post-orogenic intrusions, as probes of crust beneath the orogen, were compared to Sask craton rocks and ca. 1850 Ma orogenic rocks to infer the origin and subsurface distribution of the Sask craton within the internides of the Trans-Hudson orogen. Results show that post-orogenic intrusions within most of the Glennie Domain and Hanson Lake block were derived, at least in part, from Archean source materials, demonstrating that the Sask craton lies beneath Paleoproterozoic orogenic rocks present at the surface. In contrast, common Pb and Sm-Nd isotopic compositions from pegmatites and aplites of the La Ronge Domain are essentially identical with those of the Paleoproterozoic orogenic rocks into which they are intruded, indicating derivation by partial melting of similar rocks. Thus, if the Sask craton extended to the west beneath the La Ronge Domain, it was beneath the zone of melting that produced the post-orogenic intrusions, making it unlikely that the Sask craton is a detached part of the Hearne craton. Many samples from the Sask craton have elevated Pb-208/Pb-204 ratios, unlike Superior craton or Hearne craton rocks, suggesting that the Sask craton was derived from an exotic source, such as the Wyoming craton, which shares similar elevated Pb-208/Pb-204 ratios.

Pharmacological evaluation of an [I-123] labelled imidazopyridine-3-acetamide for the study of benzodiazepine receptors

In vitro binding of the iodinated imidazopyri dine, N',N'-dimethyl-6-methyl-(4'-[I-123]iodophenyl)imidazo[1,2-a]pyridine-3-acetamide [I-123]IZOL to benzodiazepine binding sites on brain cortex, adrenal and kidney membranes is reported. Saturation experiments showed that [I-123]IZOL, bound to a single class of binding site (n(H)=0.99) on adrenal and kidney mitochondrial membranes with a moderate affinity (K-d=30 nM). The density of binding sites was 22 +/- 6 and 1.2 +/- 0.4 pmol/mg protein on adrenal and kidney membranes, respectively. No specific binding was observed in mitochondrial-synaptosomal membranes of brain cortex. In biodistribution studies in rats, the highest uptake of [I-123]IZOL was found 30 min post injection in adrenals (7.5% ID/g), followed by heart, kidney, lung (1% ID/g) and brain (0.12% ID/g), consistent with the distribution of peripheral benzodiazepine binding sites. Pre-administration of unlabelled IZOL and the specific PBBS drugs, PK 11195 and Ro 5-4864 significantly reduced the uptake of [I-123]IZOL by 30% (p < 0.05) in olfactory bulbs and by 51-86% (p < 0.01) in kidney, lungs, heart and adrenals, while it increased by 30% to 50% (p < 0.01) in the rest of the brain and the blood. Diazepam, a mixed CBR-PBBS drug, inhibited the uptake in kidney, lungs, heart, adrenals and olfactory bulbs by 32% to 44% (p < 0.01) but with no effect on brain uptake and in blood concentration. Flumazenil, a central benzodiazepine drug and haloperidol (dopamine antagonist/sigma receptor drug) displayed no effect in [I-123]IZOL in peripheral organs and in the brain. [I-123]IZOL may deserve further development for imaging selectively peripheral benzodiazepine binding sites. (c) 2006 Elsevier Inc. All rights reserved.

Remodeling of extracellular matrix at ovulation of the bovine ovarian follicle

Using immunohistochemistry and RNA analyses we examined the fate of components of a newly identified matrix that develops between granulosa cells (focimatrix, abbreviated from focal intraepithelial matrix) and of the follicular basal lamina in ovulating bovine ovarian follicles. Pre- and postovulatory follicles were generated by treatment with estradiol (Day 1), progesterone (Days 1-10), and prostaglandin analogue (Day 9) with either no further treatment (Group 1, n = 6) and or with 25 mg porcine LH (Day 11, Group 2, n = 8 or Day 10, Group 3, n = 8) and ovariectomy on Day 12 (12-14 hr post LH in Group 2, 38-40.5 hr in Group 3). In the time frame examined no loss of follicular basal lamina laminin chains beta 2 and gamma 1 or nidogen 1 was observed. In the follicular basal lamina collagen type IV alpha 1 and perlecan were present prior to ovulation; after ovulation collagen type IV alpha 1 was discontinuously distributed and perlecan was absent. Versican in the theca interna adjacent to the follicular basal lamina in preovulatory follicles was not observed post ovulation, however, the granulosa cells then showed strong cytoplasmic staining for versican. Expression of versican isoforms V0, V1, and V3 was detected at all stages. Focimatrix was observed in preovulatory follicles. It contained collagen type IV alpha 1, laminins beta 2 and gamma 1, nidogen 1 and perlecan and underwent changes in composition similar to that of the follicular basal lamina. In conclusion focimatrix and the follicular basal lamina are degraded at ovulation. Individual components are lost at different times.

The critical dietary potassium concentration for induction of mineral disorders in non-lactating Welsh Mountain sheep

Diets with more than 30 g K/kg DM have previously been associated with hypomagnesaemia in grazing cattle, and to test whether such diets lead to mineral disorders in sheep, the absorption of Mg and other elements was investigated using experimental diets to which KC I had been added to provide 27, 29, 32 or 34 g K/kg DM. The apparent absorption, balance and apparent retention of Mg, and to a lesser extent Ca, were reduced for sheep offered the diets with 32 or 34 g K/kg DM. The absorption and retention of K, Na, P, Zn, Pb and Cd was not affected by treatment. The blood intracellular Ca concentration was reduced by the diets with 29, 32 or 34 g K/kg DM, compared to the diet with 27 g K/kg DM, but the concentration of other elements was unaffected. Blood plasma Ca concentration was increased at the highest level of K inclusion, providing evidence of mild hyperkalaemia and the involvement of Ca homeostatic mechanisms. It is concluded that Mg absorption by sheep will be impaired if the diet contains more than 30 g K/kg DM, equivalent to an intake of approximately 13 g K/d, but that a high K diet may be beneficial before parturition to accustom the sheep to Ca mobilization before lactation. (c) 2005 Elsevier B.V. All rights reserved.

Rare earth element and yttrium variability in South East Queensland waterways

We present data for the rare earth elements and yttrium (REY) in the National Research Council of Canada natural river water reference material SLRS-4 and 19 natural river waters from small catchments in South-East Queensland, Australia, by a direct ICP-MS method. The 0.22 mu m filtered river water samples show a large degree of variability in both the REY concentration, e.g., La varies from 13 to 1157 ppt, and shape of the alluvial-sediment-normalised REY patterns with different samples displaying light, middle or heavy rare earth enrichment. In addition, a spatial study was undertaken along the freshwater section of Beerburrum Creek, which demonstrates that similar to 75% of the total REYs in this waterway are removed prior to estuarine mixing without evidence of fractionation.