Twelve year analysis of aerobic-only blood cultures for routine detection of bacteraemia.

Sampling practices determine the accuracy of blood culture in diagnosing bloodstream infection. The main acute hospital in this study introduced aerobic-only routine blood cultures aiming to increase the volume and number of aerobic samples. At the smaller acute site, aerobic-anaerobic pairs were sent routinely. Culture yield and sampling practices were compared at these two sites and it was found that anaerobic cultures increased the yield of pathogens including facultative anaerobes. Volume cultured and number of samples sent fell short of national recommendations. The aerobic-only policy did not result in more blood being cultured. Based on these findings, the main acute hospital is reintroducing aerobic-anaerobic pairs for routine culture.

Effects of rimonabant, a cannabinoid CB1 receptor ligand, on energy expenditure in lean rats.

OBJECTIVE: To determine the effect of rimonabant on energy expenditure (O2 consumption) in rats at different metabolic states and in cannabinoid CB1 receptor-deficient (CB1R-/-) mice. DESIGN: Animals were exposed to light-dark cycles and fed only during dark cycles. Rimonabant or vehicle was administered together with food (absorptive), following overnight feeding (postabsorptive) or following a whole day of no food (fasting). Indirect calorimetric measurements, physical activity and food intake were measured continuously. RESULTS: Compared with vehicle-treated rats, rats administered 3 and 10 mg kg(-1) rimonabant showed an 18 and 49% increase in O2 consumption, respectively after 3 h. A second dose of rimonabant administered 9-14.5 h after the first one failed to affect O2 consumption, suggesting the development of tolerance. Similarly, stereotypic behaviors and ambulatory activity increased following the first dose but these effects were not observed after the second dose. Respiratory quotients revealed no effect of rimonabant on rates of carbohydrate and fat oxidation. Analysis of the correlation between O2 consumption and physical activity indicated that factors other than increased physical activity may contribute to the increase in O2 consumption. Similar studies in mice demonstrated that wild type but not CB1R-/- mice showed a change in O2 consumption and physical activity following rimonabant administration, suggesting that these effects are mediated by the cannabinoid CB1 receptor. CONCLUSION: Previous studies suggested that reduced food intake alone may not explain the weight reduction observed with rimonabant. Our studies suggest that rimonabant stimulates significant acute energy expenditure in non-obese rodents, which could not be completely accounted for by an increase in physical activity. However, with the observation that there is rapid development of tolerance, these results suggest that there may be additional mechanism(s) that lead to weight loss in these rodents.

Food intake in free-feeding and energy-deprived lean rats is mediated by the neuropeptide Y5 receptor.

The new neuropeptide Y (NPY) Y5 receptor antagonist CGP 71683A displayed high affinity for the cloned rat NPY Y5 subtype, but > 1, 000-fold lower affinity for the cloned rat NPY Y1, Y2, and Y4 subtypes. In LMTK cells transfected with the human NPY Y5 receptor, CGP 71683A was without intrinsic activity and antagonized NPY-induced Ca2+ transients. CGP 71683A was given intraperitoneally (dose range 1-100 mg/kg) to a series of animal models of high hypothalamic NPY levels. In lean satiated rats CGP 71683A significantly antagonized the increase in food intake induced by intracerebroventricular injection of NPY. In 24-h fasted and streptozotocin diabetic rats CGP 71683A dose-dependently inhibited food intake. During the dark phase, CGP 71683A dose-dependently inhibited food intake in free-feeding lean rats without affecting the normal pattern of food intake or inducing taste aversion. In free-feeding lean rats, intraperitoneal administration of CGP 71683A for 28 d inhibited food intake dose-dependently with a maximum reduction observed on days 3 and 4. Despite the return of food intake to control levels, body weight and the peripheral fat mass remained significantly reduced. The data demonstrate that the NPY Y5 receptor subtype plays a role in NPY-induced food intake, but also suggest that, with chronic blockade, counterregulatory mechanisms are induced to restore appetite.

Effect of divalent cation chelation on dihydropyridine binding in isolated cardiac sarcolemma vesicles.

The effect of divalent cation chelation on specific nitrendipine and ouabain binding has been determined in a highly enriched sarcolemma preparation isolated from canine ventricle. Maximal high-affinity nitrendipine binding measured in the absence of added calcium or magnesium was 997 +/- 103 fmol/mg protein. Nitrendipine binding in the presence of EDTA significantly decreased to 419 +/- 42 fmol/mg protein (P less than 0.001) which equates to 42.0% of control. The simultaneous presence of EDTA and A23187 in the binding buffer resulted in a decrease in nitrendipine binding to below detectable levels. These results suggest that divalent cations trapped within vesicles can support high affinity nitrendipine binding. Evaluation of dihydropyridine binding at various pH values suggested that the loss of binding below pH 7.0 and above pH 8.0 may result indirectly from a change in divalent cation binding rather than a direct effect on dihydropyridine binding per se. The maximal binding of ouabain determined in the presence of magnesium and inorganic phosphate averaged 340 +/- 7.4 pmol/mg protein. Pre-treatment of the preparation with sodium dodecyl sulfate (SDS) in order to express binding in sealed inside-out (IO) vesicles, increased ouabain binding to 471 +/- 27 pmol/mg protein. Thus, these preparations averaged 27.8% sealed IO vesicles. Addition of EDTA in the absence of magnesium in the binding buffer reduced ouabain binding to 204 +/- 7.7 and 11.7 +/- 3.5 pmol/mg protein in control and SDS-treated preparations, respectively. These findings suggest that this sarcolemma preparation consists of 43.6% sealed right-side-out (RO) vesicles which contain sufficient endogenous divalent cation trapped in the intravesicular space, to support ouabain binding. The correspondence between the percentage of ouabain binding that remains in the presence of EDTA and the percentage of nitrendipine binding observed under the same conditions is consistent with the hypothesis that divalent cations support nitrendipine binding by interaction with a site or sites accessible only from the cytoplasmic membrane surface and that nitrendipine and ouabain binding sites occur in the same vesicles (i.e., the nitrendipine binding site is of sarcolemma origin).

Isolation of a highly enriched sarcolemma membrane fraction from canine heart.

A highly enriched sarcolemma preparation was isolated by differential centrifugation of a canine ventricular homogenate followed by centrifugation of a membrane fraction layered over 22% (w/v) sucrose. Ouabain binding, ouabain-sensitive potassium phosphatase activity and 5'-nucleotidase activity were enriched 19--27 fold over the homogenate whereas Ca2+-ATPase and succinate dehydrogenase activities were 0.75 and 0.36, respectively, of that for the homogenate. The isolation procedure was relatively rapid and yielded about 2.0 mg protein/100 g of ventricular muscle. The highest salt concentration used in the procedure was 0.6 M KCl and no detergents were employed. Initial characterization studies suggested that the sarcolemma-enriched fraction consists predominantly if not totally of freely permeable membrane vesicles and that the sarcolemma does not manifest a Ca2+-ATPase activity, at least within the limits of the assay procedures employed. This preparation was concluded to be about 1.5- to 4-fold more highly enriched with sarcolemmal markers than preparations obtained by previously published procedures. Accordingly, the preparation provides an improved basis for the probe of calcium movements that occur across the sarcolemma in association with the excitation-contraction-relaxation sequence of the mammalian myocardial cell.

The hydraulic capacity of deteriorating sewer systems.

Sewer and wastewater systems suffer from insufficient capacity, construction flaws and pipe deterioration. Consequences are structural failures, local floods, surface erosion and pollution of receiving waters bodies. European cities spend in the order of five billion Euro per year for wastewater network rehabilitation. This amount is estimated to increase due to network ageing. The project CARE-S (Computer Aided RE-habilitation of Sewer Networks) deals with sewer and storm water networks. The final project goal is to develop integrated software, which provides the most cost-efficient system of maintenance, repair and rehabilitation of sewer networks. Decisions on investments in rehabilitation often have to be made with uncertain information about the structural condition and the hydraulic performance of a sewer system. Because of this, decision-making involves considerable risks. This paper presents the results of research focused on the study of hydraulic effects caused by failures due to temporal decline of sewer systems. Hydraulic simulations are usually carried out by running commercial models that apply, as input, default values of parameters that strongly influence results. Using CCTV inspections information as dataset to catalogue principal types of failures affecting pipes, a 3D model was used to evaluate their hydraulic consequences. The translation of failures effects in parameters values producing the same hydraulic conditions caused by failures was carried out through the comparison of laboratory experiences and 3D simulations results. Those parameters could be the input of 1D commercial models instead of the default values commonly inserted.

Intracellular Ca2+ mobilization by thyrotropin, carbachol, and adenosine triphosphate in dog thyroid cells.

The effect of TSH, carbachol (CC), and ATP on intracellular calcium concentration ([Ca2+]i) in primary cultures of dog thyroid cells was examined using the fluorescent Ca2+ indicator fura-2. TSH caused an increase in [Ca2+]i at 37 C, but not 22 C, while it increased cAMP formation in these cells at both 22 and 37 C. CC and ATP increased [Ca2+]i at both 22 and 37 C. The CC-induced increase in [Ca2+]i was under muscarinic receptor control, and it was biphasic, with an initial spike followed by a sustained increase at a lower level. TSH and ATP were weaker agonists compared to CC, since maximal doses of TSH (100-500 mU/ml) and ATP (100-500 microM) increased [Ca2+]i by 40-70% over basal levels, compared to a 2- to 4-fold increase in [Ca2+] induced by maximal doses of CC (10-50 microM). The TSH-induced increase in [Ca2+]i was transient, returning to basal levels within 1-2 min after application of the agonist. All three agents were able to transiently increase [Ca2+]i to be internal stores. In the presence of the inorganic Ca2+ channel blockers La3+, Ni2+, and Co2+, the peak [Ca2+]i change was little affected, while the persistent response to CC and ATP was blocked, indicating dependence of this phase on influx of Ca2+. Paradoxically, these channel blockers abolished the effect of TSH on [Ca2+]i. TSH stimulation of cAMP formation was also inhibited 80-90% by these blockers, but not in Ca2+-free/EGTA buffer. These results suggest that the Ca2+ channel blockers may have actions in addition to inhibition of Ca2+ entry in these cells. TMB-8 [8-(N,N-diethylamino)octyl-3,4,5-trimethoxybenzoate HCl] specifically blocked both the initial and sustained increase induced by CC, while having no effect on ATP or TSH-induced [Ca2+]i, suggesting that TMB-8 may not be a general antagonist of Ca2+ mobilization. Activators of protein kinase-C, such as phorbol esters or an analog of diacylglycerol, inhibited the [Ca2+]i rise induced by all the three agonists used, indicating a regulatory role of protein kinase-C activation on [Ca2+]i in these cells. In FRTL-5 cells, [Ca2+]i was also increased by TSH and ATP, but not by CC. ATP, however, was a more effective agonist than in dog thyroid cells, while TSH increased [Ca2+]i by a similar magnitude in both cell types. The results of the present study demonstrate that TSH, albeit of lesser potency than CC, increases [Ca2+]i by causing intracellular Ca2+ mobilization in cultured dog thyroid cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Effect of oxidant stress on calcium signaling in vascular endothelial cells.

The endothelial cell is recognized as a critical modulator of blood vessel tone and reactivity. This regulatory function of endothelial cells occurs via synthesis and release of diffusible paracrine substances which induce contraction or relaxation of adjacent vascular smooth muscle. In response to stimulation by blood-borne agonists such as bradykinin or histamine, the endothelial cell utilizes cytosolic ionic Ca2+ as a trigger in the transduction of the stimulatory signal into a paracrine response. Considerable evidence has accumulated to indicate that various forms of biologically important oxidant stress alter vascular function in an endothelium-dependent manner. Further, oxidant stress is known to alter the mechanisms which govern Ca2+ homeostasis in the endothelial cell. Recently, we have described a model in which the oxidant tert-butylhydroperoxide is utilized to examine the effects of oxidant stress on Ca(2+)-dependent signal transduction in vascular endothelial cells. In this model, three temporal phases are evident and consist of (1) inhibition of the agonist-stimulated Ca2+ influx pathway, (2) inhibition of receptor-activated release of Ca2+ from internal stores and elevation of resting cytosolic free Ca2+ concentration, and (3) progressive increase in resting cytosolic Ca2+ concentration and loss of responsiveness to agonist stimulation. In this review, the mechanisms which characterize agonist-stimulated Ca2+ signaling in vascular endothelial cells, and the effects of oxidant stress on signal transduction will be described. The mechanisms potentially responsible for oxidant-induced inhibition of Ca2+ signaling will be considered.

Concomitant and hormonally regulated expression of trp genes in bovine aortic endothelial cells.

Recent findings have suggested that the vertebrate trp family of channel proteins is the structural basis for Ca2+ influx through the capacitative calcium entry (CCE) pathway. We have discerned, in bovine aortic endothelial cells, the concomitant expression of four such vertebrate genes: trp-1 (two splice variants), trp-3, trp-4 and trp-5. Exogenous hormones rendered dynamic effects on the transcript levels of these genes. Most notably, beta-estradiol significantly down-regulated trp-4 while trans-retinoic acid dramatically up-regulated trp-5; yet these hormones rendered little change in CCE. These findings suggest that the extent of a given trp channel's participation in CCE is not reflected in alterations of its transcript level.

NKP608: a selective NK-1 receptor antagonist with anxiolytic-like effects in the social interaction and social exploration test in rats.

NKP608 is a non-peptidic derivative of 4-aminopiperidine which acts as a selective, specific and potent antagonist at the neurokinin-1 (NK-1) receptor both in vitro and in vivo. In vitro, the binding of NKP608 to bovine retina was characterized by an IC50 of 2.6+/-0.4 nM, whereas the compound's affinity to other receptor binding sites, including NK-2 and NK-3, was much lower. Species differences in IC(50) values with NKP608 were less pronounced than with previously described NK-1 receptor antagonists, being 13+/-2 and 27+/-2 nM in gerbil midbrain and rat striatum, respectively. In vivo, using the hind foot thumping model in gerbils, NKP608 exhibited a potent NK-1 antagonistic activity following oral administration (ID(50)=0.23 mg/kg; 2 h pretreatment), supporting a central activity of NKP608. The compound had a long duration of action with an ID(50) value of 0. 15 mg/kg p.o. and 0.38 mg/kg p.o. following a pretreatment of 5 and 24 h, respectively. Following a subchronic administration for 7 consecutive days (once daily) there was no evidence for the development of tolerance or accumulation. In the social interaction test performed in a highly illuminated, unfamiliar test arena, NKP608 specifically increased the time the two rats spent in social contact, and there was no concomitant increase in parameters reflecting general activity, i.e. ambulation (number of square entries) or the number of rearings. Active social time was maximally increased at a dose range of 0.01-1 mg/kg p.o. NKP608, the effect being weaker or absent at both lower (0.001 mg/kg p.o.) and higher (10 mg/kg p.o.) doses. A comparable bell-shaped dose-response relation was seen in the social exploration test in rats. In this modified resident/intruder paradigm, maximal increase in social contact of the intruder rat directed towards the resident rat was seen at a similar dose range (0.03-3 mg/kg p.o.) The effects observed following an acute oral administration of NKP608 were comparable to those seen following a treatment with the well-known benzodiazepine, chlordiazepoxide, in both these tests. These findings indicate that NKP608 exhibits an anxiolytic-like effect and that this effect, as concluded from the observed antagonism of the hind foot thumping induced by i.c.v. administration of the NK-1 receptor agonist SPOMe, is centrally mediated. This makes this compound a potentially promising candidate for treating anxiety-related disorders in humans.

Endothelium-dependent relaxant effect of neurokinins on rabbit aorta is mediated by the NK1 receptor.

Several neurokinins, namely substance P, neurokinin A, neurokinin B, [beta-Ala8]neurokinin A-(4-10) and senktide, were tested on noradrenaline-precontracted rabbit aortic rings to characterize the receptor mediating their endothelium-dependent relaxant effect in this preparation. CP-96,345, the new nonpeptide antagonist selective for the NK1 receptor, was also studied. Substance P, neurokinin A and neurokinin B, in that order of potency, were effective in relaxing precontracted rings, indicating the involvement of the NK1 receptor; [beta-Ala8]neurokinin A-(4-10) and senktide, which are selective agonists for NK2 and NK3 receptors, respectively, had no significant relaxant effect. The relaxant effects of substance P, neurokinin A and neurokinin B were competitively antagonized by nanomolar concentrations of CP-96,345. These findings support the view that the NK1 receptor mediates the endothelium-dependent relaxant effect of the neurokinins in rabbit aorta.

Gluten-free diet in patients with dermatitis herpetiformis. Effect on the occurrence of antibodies to reticulin and gluten.

In a prospective study of 51 patients with dermatitis herpetiformis (DH), the occurrence of antibodies against reticulin and gluten was investigated and the influences of gluten-free, gluten-restricted, and normal diets on the persistence of these antibodies were compared. After a period extending from 11 to 47 months, none of the patients who were eating a gluten-free diet had reticulin or gluten antibodies. The patients eating a normal diet had the same incidence of reticulin autoantibodies at the end of the study as at its beginning, and gluten antibodies developed in some additional patients during the follow-up period. Thus, we have shown that the elimination of gluten from the diet of patients with DH has an important influence on the occurrence of antibodies to both reticulin and gluten.

Maitotoxin-induced membrane blebbing and cell death in bovine aortic endothelial cells.

BACKGROUND: Maitotoxin, a potent cytolytic agent, causes an increase in cytosolic free Ca2+ concentration ([Ca2+]i) via activation of Ca2+-permeable, non-selective cation channels (CaNSC). Channel activation is followed by formation of large endogenous pores that allow ethidium and propidium-based vital dyes to enter the cell. Although activation of these cytolytic/oncotic pores, or COP, precedes release of lactate dehydrogenase, an indication of oncotic cell death, the relationship between CaNSC, COP, membrane lysis, and the associated changes in cell morphology has not been clearly defined. In the present study, the effect maitotoxin on [Ca2+]i, vital dye uptake, lactate dehydrogenase release, and membrane blebbing was examined in bovine aortic endothelial cells. RESULTS: Maitotoxin produced a concentration-dependent increase in [Ca2+]i followed by a biphasic uptake of ethidium. Comparison of ethidium (Mw 314 Da), YO-PRO-1 (Mw 375 Da), and POPO-3 (Mw 715 Da) showed that the rate of dye uptake during the first phase was inversely proportional to molecular weight, whereas the second phase appeared to be all-or-nothing. The second phase of dye uptake correlated in time with the release of lactate dehydrogenase. Uptake of vital dyes at the single cell level, determined by time-lapse videomicroscopy, was also biphasic. The first phase was associated with formation of small membrane blebs, whereas the second phase was associated with dramatic bleb dilation. CONCLUSIONS: These results suggest that maitotoxin-induced Ca2+ influx in bovine aortic endothelial cells is followed by activation of COP. COP formation is associated with controlled membrane blebbing which ultimately gives rise to uncontrolled bleb dilation, lactate dehydrogenase release, and oncotic cell death.

Sanglifehrins A, B, C and D, novel cyclophilin-binding compounds isolated from Streptomyces sp. A92-308110. II. Structure elucidation, stereochemistry and physico-chemical properties.

A novel class of macrolides, the sanglifehrins, was discovered by screening of actinomycete strains with a cyclophilin-binding assay. The chemical structures and absolute stereochemistries of the sanglifehrins A, B, C and D were determined unambiguously by NMR-techniques and by X-ray crystallography of the complex with cyclophilin A. Sanglifehrin A consists of a 22-membered macrocycle containing a tripeptide subunit and features in position 23 a chain of nine carbon atoms bearing a spirocyclic substituent. Sanglifehrins A and B are genuine metabolites whereas sanglifehrins C and D are artefacts.

Antascomicins A, B, C, D and E. Novel FKBP12 binding compounds from a Micromonospora strain.

5 novel ascomycin-like compounds, antascomicins A, B, C, D and E were isolated from a strain of Micromonospora. The antascomicins bind strongly to the FK506-binding protein FKBP12 and antagonize the immunosuppressive activity of FK506 and rapamycin. The strain description, fermentation, structure elucidation and biological activity of these compounds are described.

Cymbimicin A and B, two novel cyclophilin-binding structures isolated from actinomycetes.

Two novel metabolites, cymbimicins A and B, were isolated from the culture broth of a strain of Micromonospora sp. by screening for cyclophilin binding metabolites from actinomycete strains. Cymbimicin A binds to cyclophilin A with a high affinity six fold lower than to that of cyclosporin A. The binding affinity of cymbimicin B is about 100 times lower. The taxonomy of the producing strain, fermentation, isolation, physical and biological properties and structure elucidation are described.

Sanglifehrins A, B, C and D, novel cyclophilin-binding compounds isolated from Streptomyces sp. A92-308110. I. Taxonomy, fermentation, isolation and biological activity.

A novel class of macrolides for which the name sanglifehrins is proposed, has been discovered from actinomycete strains based on their high affinity binding for cyclophilin A (CypA), an immunophilin originally identified as a cytosolic protein binding cyclosporin A (CsA). The sanglifehrins were produced by Streptomyces sp. A92-308110. They were isolated and purified by extraction and several chromatographic, activity-guided steps. Sanglifehrins A and B exhibit a 10 to approximately 20 fold higher affinity for CypA than CsA, whereas the affinity of sanglifehrins C and D for CypA is comparable to that of CsA. Sanglifehrins exhibit a lower immunosuppressive activity than CsA when tested in the mixed lymphocyte reaction. Their in vitro activity indicates that they belong to a novel class of immunosuppressants.

[Monitoring of medication in optimalization of preoral theophylline in long-term care]. / Uberwachung der Medikation bei der Optimierung der peroralen Theophyllin-Langzeitbehandlung.

A narrow therapeutic index, considerable interpatients variability of clearance and biotransformation, rapid alterations of pharmacokinetics point out theophylline to a problem-drug. An effective long-term treatment with this drug is only possible with retard-preparations. For a certain therapy a drug monitoring-controlling the theophylline-concentration in the serum--is necessary, in what extend and under which conditions is discussed in detail.

A software monitor for intermittent bacteria contamination in urban rivers.

Norwegian receiving waters are of such high water quality that authorities consider opening them for bathing. The leading parameter to monitor the quality of bathing waters is fecal coliform bacteria (FC). For this parameter no rapid detection method is available. The main objective of this case study was to find a way to quickly predict bacteria contamination by observing different online parameters such as flow, conductivity or spectral absorption coefficient (SAC). In this study historical data from 1994 to 2000 was analyzed, and over a period of five weeks water samples were taken and analyzed for bacteria. The analysis of the historical data revealed fundamental sampling problems, which made the data useless for the purpose of this study. The analysis of the data collected for this study showed that exceeding the bathing water standard for bacteria can be predicted by evaluating the SAC with an acceptable accuracy. Furthermore a simple river quality model was implemented, including bacteria as a load fraction. With the help of rain data and discharge predictions expected bacteria numbers exceeding the bathing water standard could also be forecast.

Urban drainage redefined: from stormwater removal to integrated management.

Even though urban drainage has been practised for more than 5000 years, many challenges arising from growing demands on drainage still remain with respect to runoff quantity and quality; landscape aesthetics, ecology and beneficial uses; and operation of existing urban wastewater systems. Further advances can be achieved by adopting an integrated approach, optimal operation of the existing infrastructure, advanced pollution and runoff source controls, improved resilience of receiving waters, and adaptive water management. The specific research needs include new technologies and strategies for stormwater management, advanced treatment of urban wet-weather effluents, and tools for analysis and operation of drainage systems. High diversity of demands on, and region/site specific conditions of, urban drainage shapes the role of urban drainage experts--as mediators among the many stakeholders and fields involved.