Primary HHV 6 infection after liver transplantation with acute graft rejection and multi-organ failure: successful treatment with a 2.5-fold dose of cidofovir and reduction of immunosuppression.

HHV type 6 has been reported with enhanced pathogenicity in immunocompromised patients. Herein, we report about a two-yr-old girl who experienced primary HHV 6 infection after liver transplantation. She clinically presented with graft rejection and necrotic hepatitis as well as high fever, pneumonitis with respiratory failure and a rash. Therapy with cidofovir of 5 mg/kg per wk did not show improvement, so that a full pharmacokinetic profile of cidofovir was performed. It demonstrated enhanced body weight normalized clearance of cidofovir and cidofovir dosage was augmented to 12 mg/kg per wk to reach adequate drug exposure. With additional reduction of immunosuppression, the patient dramatically improved and liver function stabilized.

Evaluation of the users value of salts against apple scab and powdery mildew for the integrated fruit production.

As new fungicides are mainly unisite action fungicides, the problem of fungicide resistance development is becoming more important every year. Combining chemical fungicides, which is the best anti-resistance strategy, is not always possible or recommended in the case when the number of available chemical fungicides are limited or a reduction in fungicide use is asked for. Therefore the use of salts as an anti-resistance strategy was looked upon. The salts evaluated were K(HCO3), KH2PO3, KHPO4 and K2SiO3. When using these salts as an anti-resistance strategy the efficacy obtained when spraying the compounds alone was often to low to be used in rotation with chemical fungicides. Only with K(HCO3)2 a good efficacy can be observed in some years. The variation in efficacy with K(HCO3)2 observed is higher for powdery mildew. Chitosan was also included in the trials against powdery mildew, however chitosan had no effect on the infestation.

Brown spot control on pear: infection models versus the inoculum pressure in Belgium.

Stemphylium conidia and Pleospora ascospores were monitored in pear orchards in the region of Haspengouw in Belgium during the infectious periods of brown spot disease (end May - end August) in the years 2002, 2004 and 2005. The seasonal and daily dynamics of the captured fungal spores are discussed and a correlation analysis was performed to determine possible correlation with weather parameters. Furthermore the fungicide cover obtained by spraying upon climatological infection risk is compared with the inoculum pressure in the orchards. Pleospora ascospores were found until the end of May and discharge took place during rain events at the same time-points when Venturia inaequalis ascospores were detected. Although the fungi ejected during the same time period, the relative importance of the different spore peaks differed. The first conidia were detected at the end of May. After that date conidia were found almost every day. Seasonal dynamics of the conidia clearly differed between the years and also the number of spores retrieved differed. No spores were found below 7.5 degrees C and only at a temperature above 12.5 degrees C an increase in conidia was observed. When looking at the daily dynamics, a significant negative correlation was found between the aerial spore concentration and relative humidity and leaf wetness, and a significant positive correlation with wind, temperature and water vapor pressure deficit. When spraying upon a BSP-cast CR threshold value of 0.4, all days with spore discharges above 3 conidia/m3/day were covered in 2002 and 2004. In 2005, a year with very low infection risk and infestation, two periods with high spore discharge were not covered. The observations made show that incorporation of the inoculum pressure into the infection models will probably not lead to a big improvement of the infection models.

Botrytis infection warnings in strawberry: reduced enhanced chemical control.

The fungal pathogen Botrytis cinerea is the causal agent of grey mould, the most important fungal fruit rot disease in strawberry in Europe. Currently disease control for grey mould is based on preventive spraying every five to seven days during flowering and harvest. Replacing preventive spraying with applications based on infection warnings can optimize performance and reduce the amount of sprays needed. Success of this approach will depend on the accuracy of the model used to predict disease outbreak. For this reason three infection models (BOTEM, BoWaS, DSS-Italy) were evaluated during the growth seasons of 2003 and 2004. The experiments included June bearing, retarded June bearing and ever bearing strawberries. In all experiments the use of infection models leaded to a reduced number of fungicide applications. However the efficacy of the different models towards the control of B. cinerea also decreased compared to the efficacy obtained with a standard 7 day schedule. Best results were obtained with BOTEM, developed by HRI (Horticultural Research International, East-Malling, UK): 17-60% reduction in fungicide use and an efficacy between 66-93 depending on the growth season, culture practice and the fungicides used. Compared with routine preventive spraying, the Botrytis fruit rot percentage is slightly higher. A higher efficacy with Botrytis infection warnings can only be obtained if infection warnings change from curative to preventive. A retroactive evaluation of a preventive warning system was included. Making use of the 48h weather forecasts supplied by the Royal Meteorological Institute of Belgium (KMI) based on ALADIN for the region of Haspengouw, it was possible to replace 30 up to 100% of the curative application by preventive spraying depending on the experiment and the threshold set for the preventive model.

Receptor mechanisms involved in the 5-HT-induced inotropic action in the rat isolated atrium.

1. The effects of 5-hydroxytryptamine (5-HT) in rat cardiac preparations were studied. 5-HT up to 10 microM failed to affect contractility in papillary muscles. However, in electrically driven (1 Hz) left atria 5-HT exerted a positive inotropic effect that started at 1 microM and attained its maximum at 10 microM (312+/-50% of predrug value, n=8). 2. 5-HT 10 microM stimulated the content of inositol-1,4,5-trisphosphate but not of cyclic AMP in rat left atria. 3. Plasma and serum levels of 5-HT amounted to about 0.3 microM and 15 microM, respectively. 4. The selective 5-HT4 receptor antagonists GR 125487 (10 nM and 1 microM) and SB 203186 (1 microM) did not attenuate the positive inotropic effect of 5-HT in rat left atria. In contrast, the 5-HT2 receptor antagonist ketanserin (5 nM, 50 nM, 1 microM) resulted in a concentration-dependent diminution of the positive inotropic effect of 5-HT in rat left atria. 5. Reverse transcriptase polymerase chain reaction with specific primers detected mRNA of the 5-HT2A receptor in rat atria and ventricles, while expression of the 5-HT4 receptor was confined to atria. 6. It is suggested that the positive inotropic effect of 5-HT in electrically driven rat left atria is mediated by ketanserin-sensitive 5-HT2A receptors and not through 5-HT4 receptors.

Inositol-1,4,5-trisphosphate increase by diadenosine tetraphosphate in preparations from failing human myocardium.

In human ventricular trabeculae carneae 100 microM AP4A (diadenosine tetraphosphate) increased force of contraction to 162.8+/-15.7% of predrug value (n=9). This positive inotropic effect was accompanied by a prolongation of time parameters: time to peak tension and time of relaxation were prolonged by 7.8+/-1.3% and 14.9+/-3.8%, respectively (P<0.05). In the same trabeculae, AP4A increased IP3 (inositol-1,4,5-trisphosphate) content from 9.0+/-1.3 pmol/mg to 22.9+/-5.4 pmol/mg protein (n=5-9). In conclusion, the positive inotropic effect of AP4A in the human myocardium is likely due to an increase of IP3 mediated probably via Gq-coupled P2Y-purinoceptors. Because of the prominent role of Gq in the development of cardiac disease, these findings may lay the ground to further investigate the possible role of AP4A and/or related ligands (e.g. AP2A and AP3A) in heart failure.

The technique of endobronchial lidocaine administration does not influence plasma concentration profiles and pharmacokinetic parameters in humans.

This study investigated plasma concentration profiles, pharmacokinetic characteristics and side-effects of lidocaine following 3 different administration techniques. Sixty ASA I/II patients undergoing elective ENT-operations were randomised into 4 groups. Lidocaine 1% (1 mg/kg) was administered 50 min before the end of the operation, via a regular endotracheal tube (group 1), a suction-catheter deep endobronchially (group 2), or an EDGAR-(Endobronchial-Drug and Gas Application during Resuscitation)-tube characterized by a separate injection channel ending at the orifice of the tube (group 3). For the control group, a regular endotracheal tube was inserted without lidocaine administration (group 4). Anesthesia was induced with propofol (2 mg/kg), sufentanil (0.5 micrograms/kg), and vecuronium (0.08 mg/kg) and continued as total intravenous anesthesia with propofol (8 mg/kg/h) and oxygen in air (FiO2 = 0.33). A control and 13 blood samples were taken up to 180 min after lidocaine administration. Lidocaine plasma concentrations were determined using a fluorescence polarization immunoassay (TDxFLx). Heart rate, blood pressure, endtidal PcO2, and oxygen saturation were similar in all groups investigated. Ventilation was interrupted for 3.6 +/- 0.5 s in group 1 and 10.2 +/- 0.8 s in group 2, to administer lidocaine. Patients from group 3 were ventilated continuously because of a separate injection channel integrated in the EDGAR-tube. Sore throat was significantly increased in group 2 as compared with groups 1, 3 and 4. Asorption of lidocaine in groups 1-3 resulted in maximal mean plasma concentrations ranging from 0.78 to 0.85 micrograms/ml after 16.9 to 22.4 min.(ABSTRACT TRUNCATED AT 250 WORDS)

Interactions between bilirubin and reactive oxygen species in liver microsomes and in human neutrophil granulocytes.

Bilirubin protects polyunsaturated fatty acids from lipid peroxidation, thus preventing damage by reactive oxygen species to cell membranes and proteins. On the other hand, such reactive oxygen species may contribute to the degradation and elimination of bilirubin. We therefore examined the interactions between bilirubin and reactive oxygen species. Bilirubin is decomposed in microsomes via a NADPH-independent process. This reaction appears to be mediated by H2O2 or by the hydroxyl radical since it is stimulated by exogenous H2O2 and by cytochrome P450 inducers, which increase H2O2 production in microsomes, and is inhibited by the hydroxyl radical scavenger sodium benzoate. These results suggest that cytochrome P450 may act as a peroxidase or as a Fenton catalyst in bilirubin degradation. On the other hand, bilirubin inhibits the NADPH consumption of microsomes as well as the NADPH oxidase activity of human neutrophil granulocytes and the resulting superoxide formation in these cells. This effect on superoxide concentration may be partially due to direct interaction between superoxide and bilirubin, since bilirubin reduces the superoxide concentration in a xanthine oxidase system. Bilirubin degradation is inhibited by superoxide dismutase suggesting that bilirubin may be oxidized in this system by the superoxide radical. The bilirubin-induced reduction in superoxide concentration in the supernatant of granulocytes suggests that hyperbilirubinemia may compromise immune function.

Determination of carvedilol in human cardiac tissue by high-performance liquid chromatography.

A new high-performance liquid chromatographic method has been developed for the determination of the beta-receptor blocker carvedilol in human cardiac tissue. After homogenizing tissue samples in a microdismembrator, carvedilol and the internal standard naftopidil are extracted with acetone. The extract is evaporated to dryness and reconstituted in a potassium acetate buffer of pH 3.5. Samples are cleaned up with solid-phase extraction columns. Carvedilol and the internal standard show recoveries of 69.8 +/- 12.2% and 63.9 +/- 9.34%, respectively. The linearity range for carvedilol is 0.01-0.35 ng/mg (parts per billion) tissue (wet weight), and the limit of quantitation is 0.01 ng/mg. The percentage coefficient of variation of the intra-assay varies between 1.45 and 5.38% and the interassay between 4.25 and 6.96%. To use as an application of the assay, the cardiac carvedilol tissue level in a patient on oral carvedilol therapy for congestive heart failure is reported.

Quantitation of diltiazem in human cardiac tissue using high-performance liquid chromatography.

A new high-performance liquid chromatographic method has been developed for the determination of diltiazem in human cardiac tissue. Tissue samples are homogenized and digested with trypsin solution. Diltiazem and the internal standard are extracted with acetone. The extract is evaporated to dryness and reconstituted in potassium phosphate buffer. Samples are then cleaned up with solid-phase extraction columns. Diltiazem and the internal standard show recoveries of 59% +/- 16 and 52% +/- 13. The linearity range is 0.12-2.25 ng/mg wet weight. The limit of quantitation is 0.12 ng/mg (w/w). The percentage coefficient of variation of intra-assay varies between 3.57 and 11.2, and that of interassay varies between 5.42 and 11.7. As an application of the assay, a diltiazem cardiac tissue level in a patient on oral therapy for supraventricular tachycardia is reported.

Long-term availability of Tc deposited on soil after accidental releases.

Considerable uncertainties are associated with the transfer of Tc from soil to plant. Mathematical models built to simulate that transfer generally use a single value derived from short-term experiments. Recent observations have, however, given a clear indication that the soil-to-plant transfer varies with time, depending on the biogeochemical cycling of this element. The long-term behavior of Tc in soils accidentally contaminated has been studied for 4 y in lysimeters exposed to natural climatic conditions. An important fraction of the deposit (70%) is rapidly removed from the 20-cm plow layer with a half-time of 54 +/- 2 d; the remaining fraction tends to persist in the soil for long periods of time (half-time = 30 +/- 16 y). Immediately after the deposit, the plant-to-soil concentration ratios are very high (about 400); they decrease to six 3 y after the contamination. The shape of the evolution of the contamination measured in grass after an accidental deposit is best-fitted to a sum of two exponential functions; the environmental half-times estimated for the two components are about 30 d and 2 to 3 y, respectively.

[Morphological and functional features of cytostatic drug resistance and the effects of MDR modulators]. / Morphologische und funktionelle Befunde zur Zytostatika-Resistenz und der Wirksamkeit von MDR-Modulatoren.

Manifold mechanisms of resistance can be expressed by malignancies. Profound information on this aspect is a prerequisite for comprehensive individual chemotherapy. Based on both morphological and functional findings, the diagnosis of P-Glycoprotein (P-Gp) mediated Multidrug Resistance (MDR) can be verified. In order to describe minimum criteria for conclusive diagnosis, morphological and biochemical findings (Immunocytochemistry, RT-PCR, ultrastructural and Laser-Scan microscopy) and functional data (cytostatic drug transport, proliferation) were correlated in tumor cell lines of the MDR phenotype as opposed to cells with atypical resistance. Frequently, single features of MDR are found in atypical, P-Gp negative resistance. Accumulation deficits for mitoxantrone based on vesicular drug transport were found in P-Gp negative gastric carcinoma cell line EPG85-257RNOV. Nocodazole blocked microtubule formation which is essential for vesicle transfer from perinuclear regions to the periphery of the cytosol. Cytochalasin blocked exocytosis of drug containing vesicles. MDR modulators were ineffective. Alternatively, P-Gp mediated drug extrusion and exocytosis of drug containing vesicles may constitute complementary mechanisms of resistance. In gastric carcinoma cells EPG85-257DAU, MDR modulators do increase cytosolic daunorubicin load, but drug binding to nuclear target sites is still inhibited due to drug containment within vesicles. To complicate matters, MDR modulators may be functional even in MDR negative cells, as shown in a panel of melanoma cell lines. Data show that conclusive diagnosis of P-Gp-mediated MDR should be based on more than one experimental approach including immunocytochemistry, a sensitive assay such as RT-PCR and--whenever feasible--a functional test.

Signum, a new fungicide with interesting properties in resistance management of fungal diseases in strawberries.

Signum, a new fungicide developed by BASF, was applied during 6 successive years against fungal diseases in strawberries. The product is formulated as a water dispersible granule, containing 6.7 % pyraclostrobin and 26.7 % boscalid. Pyraclostrobin is similar in chemistry to other strobilurin fungicides like kresoxim-methyl and trifloxystrobin, registered for fruit disease control. Boscalid belongs to the class of carboxyanilides. Both components in the premix formulation combine two different biochemical modes of action in the fungal cell respiration. Therefore, this co-formulation gives a broad-spectrum activity and also a reduced resistance risk for different target pathogens. Botrytis cinerea is the most important disease on strawberry-fruits and thus the control of fruit rot is mainly focused on this fungus. In average over 6 years, Signum has not only given a very good control against Botrytis fruit rot, but it has also shown a high performance in the control of Colletotrichum. Besides, Signum provides good control of powdery mildew (Podosphaera aphanis) and limits the shift to other fruit rots like leather rot (Phytophthora cactorum and leak (Rhizopus, Mucor). The availability of several categories of fungicide families with a different mode of action gives opportunities in alternating different fungicides and is the best guarantee for a sustainable control of fruit rot in all kinds of strawberry production methods. Signum should be integrated in an overall disease management program. Trials, in which the applications of Signum were timed on disease forecasting, based on environmental factors favorable for Botrytis development, were very promising. This tool can also help in establishing the IPM-concept in the production of strawberries.

Physiologically based pharmacokinetic (PBPK) modeling in children.

This review summarizes the present status of physiologically based pharmacokinetic (PBPK) modeling and simulation (M&S) and its application in support of pediatric drug research. We address the reasons that PBPK is suited to the current needs of pediatric drug development and pharmacotherapy in light of the evolution in pediatric PBPK methodologies and approaches, which were originally developed for the purpose of toxicologic evaluation. Also discussed is the current degree of confidence in using PBPK to support pediatric drug development and registration and the key factors essential for robust results and broader adoption of pediatric PBPK M&S.

Utility of N-terminal brain natriuretic peptide plasma concentrations in comparison to lactate and troponin in children with congenital heart disease following open-heart surgery.

We conducted a prospective study in a pediatric cardiac intensive care unit in order to determine the diagnostic value of N-terminal brain natriuretic peptide (N-BNP) plasma concentration in the perioperative care of children with congenital heart disease (CHD). N-BNP plasma concentrations were determined by using a validated enzyme immunoassay. We measured N-BNP the day before surgery and up to 15 days postoperatively in 23 children (age range, 0.25-11 years) undergoing cardiac surgery due to various CHDs. Supply and duration of catecholamines, vasodilators, and respiratory therapy were determined and correlated to N-BNP. In addition, troponin T (TnT) and arterial Lactat (aL) concentrations were measured simultaneously. We found a significant correlation between preoperative and maximal N-BNP levels and dosage of vasodilators (r = 0.41, p < 0.02 and r = 0.83, p < 0.01, respectively). Maximal TnT and aL levels were not correlated to dosage of vasodilators. The dosage and duration of catecholamines, the duration of respiratory therapy, and the plasma concentration of TnT and aL were not correlated to pre- or perioperative N-BNP. Maximal TnT and aL levels were correlated to duration (r = 0.53, p < 0.01 and r = 0.48, p < 0.02) and dosage (r = 0.52, p < 0.02 and r = 0.60, p < 0.01) of catecholamines and duration of respiratory therapy (r = 0.57, p < 0.01 and r = 0.50, p < 0.02). As recent studies show, N-BNP appears to be a powerful neurohumoral indicator of ventricular function and prognosis for guiding therapy in the outpatient department or for discriminating cardiac from noncardiac symptoms. In contrast, the value of N-BNP for guiding perioperative therapy in pediatric cardiac intensive care units is limited.

Plasma concentrations of N-terminal brain natriuretic peptide in healthy children, adolescents, and young adults: effect of age and gender.

Children with congenital heart disease need adequate diagnostic classification regarding their cardiovascular status (CVS). N-terminal brain natriuretic peptide (N-BNP) plasma concentration indicates dysfunction of the cardiovascular system and guides decisions concerning treatment and prognosis. Reference values are established for adults, with age-dependent increasing values and higher values in women. To avoid misclassification concerning the CVS, a large group of healthy children and adolescents can be used show the relationship between gender, age, and N-BNP and these can serve as reference values. N-BNP was measured in 434 healthy subjects (240 female and 194 male) with ages ranging from 0 to 32 years without any cardiovascular disease or renal or hepatic impairment. Measurements were performed with an electrochemiluminescence immunoassay from Roche Diagnostics. Mean N-BNP decreased from 12.6 fmol/ml (0-9 years; n = 79) to 9.41 fmol/ml (10-14 years; n = 154) and in adolescents from 6.1 (15-19 years; n = 99) to 4.8 fmol/ml (> 19 years; n = 102) in adults (p < 0.05). Mean N-BNP concerning gender did not differ in any age group younger than 19 years. In contrast, the adult female group had 78% higher N-BNP compared to the male group (p < 0.05). There was a significant peak in N-BNP at the age of 12-14 years. This study shows that reference values for N-BNP differed profoundly in children compared to adults and were up to 260% higher in children without any gender difference. Therefore, these reference values will help to avoid CVS misclassification in children for the biomarker N-BNP.

Small blood volumes from children for quantitative sotalol determination using high-performance liquid chromatography.

A sensitive high-performance liquid chromatographic method using fluorescence detection has been developed for sotalol determination in small plasma samples of children and newborns with limited blood volume. In sample sizes of 100 microl of plasma, sotalol was extracted using an internal standard and solid-phase extraction columns. Chromatographic separation was performed on a Spherisorb C6 column of 150x4.6 mm I.D. and 5 microm particle size at ambient temperature. The mobile phase consisted of acetonitrile-15 mM potassium phosphate buffer (pH 3.0) (70:30, v/v). The excitation wavelength was set at 235 nm, emission at 300 nm. The flow-rate was 1 ml/min. Sotalol and the internal standard atenolol showed recoveries of 107+/-8.9 and 97+/-8.1%, respectively. The linearity range for sotalol was between 0.07 and 5.75 microg/ml, the limit of quantitation 0.09 microg/ml. Precision values expressed as percent relative standard deviation of intra-assay varied between 0.6 and 13.6%, that of inter-assay between 2.4 and 14.4%. Accuracy varied between 86.1 and 109.8% (intra-assay) and 95.4 and 103.3% (inter-assay). Other clinically used antiarrhythmic drugs did not interfere. As an application of the assay, sotalol plasma concentrations in a 6-year-old child with supraventricular tachycardia treated with oral sotalol (3.2 mg/kg per day) are reported.