Vaccination of allogeneic haematopoietic stem cell transplant recipients: report from the international consensus conference on clinical practice in chronic GVHD

Patients lose protective immunity to vaccine-preventable diseases after haematopoietic stem cell transplantation (HSCT). Therefore, revaccination of HSCT recipients represents an important strategy for reducing morbidity and mortality associated with these infections. Since there is little consensus on vaccine recommendations and practices for allogeneic HSCT recipients with active chronic graft-versus-host disease (GVHD) the German-Austrian-Swiss-Consensus Conference on Clinical Practice in Chronic GVHD developed an immunization schedule with the aim to provide optimal patient care. The proposed vaccine recommendations include immunization against Haemophilus influenzae type b, pertussis, pneumococci, meningococci, tetanus, diphtheria, hepatitis A and B, measles, mumps and rubella, influenza, poliomyelitis, varicella-zoster virus, human papilloma virus, and tick-borne encephalitis with a particular focus on vaccination of patients with active chronic GVHD.     

Influence of treatment duration with a 600-mg dose of clopidogrel before percutaneous coronary revascularization

OBJECTIVES: We examined clinical outcomes in the Intracoronary Stenting and Antithrombotic Regimen-Rapid Early Action for Coronary Treatment (ISAR-REACT) trial based on the duration of pretreatment with a 600-mg loading dose of clopidogrel. BACKGROUND: The influence of the treatment duration with a 600-mg dose of clopidogrel before percutaneous coronary revascularization on early outcomes remains uncertain. METHODS: Among 2,159 patients with coronary disease who underwent percutaneous coronary intervention (PCI) in the ISAR-REACT trial, we examined clinical outcomes relative to the duration of pretreatment with a 600-mg dose of clopidogrel: (2 to 3 h, 3 to 6 h, 6 to 12 h, or >12 h). Patients were randomly assigned to adjunctive therapy with abciximab or placebo at the beginning of the study. The primary end point was a composite of death, myocardial infarction, or urgent revascularization within 30 days after randomization. RESULTS: No significant differences were observed between patient groups regarding the duration of pretreatment, irrespective of assignment to abciximab or placebo (p = 0.27 for interaction among abciximab/clopidogrel and placebo/clopidogrel treatment at each time interval). Occurrence of major bleeding also did not differ according to time of initial clopidogrel dosing. CONCLUSIONS: For low-to-intermediate risk patients treated with a 600-mg loading dose of clopidogrel before PCI, incremental clinical benefit within the first 30 days from durations of pretreatment >2 to 3 h was not evident.     

Overcoming compound interference in fluorescence polarization-based kinase assays using far-red tracers

Kinase-mediated phosphorylation of proteins is critical to the regulation of many biological processes, including cell growth, apoptosis, and differentiation. Because of the central role that kinases play in processes that can lead to disease states, the targeting of kinases with small-molecule inhibitors is a validated strategy for therapeutic intervention. Classic methods for assaying kinases include nonhomogenous enzyme-linked immunosorbent assays or scintillation-based formats using [gamma-(32)P]ATP. However, homogenous fluorescence-based assays have gained in popularity in recent years due to decreased costs in reagent usage through miniaturization, increased throughput, and avoidance of regulatory costs associated with the use of radiation. Whereas the readout signal from a nonhomogenous or radioactive assay is largely impervious to interferences from matrix components (such as library compounds), all homogenous fluorescent assay formats are subject to such interferences. Interference from intrinsically fluorescent compounds or from scattered light due to precipitated compounds can interfere with assays that depend on a fluorescence intensity (or fluorescence quenching), fluorescence resonance energy transfer, or fluorescence polarization-based readout. Because these interfering factors show a greater effect at lower wavelengths, one strategy to overcome such interferences is to develop fluorescent assays using longer wavelength (red-shifted) fluorescent probes. In this article, we describe the PanVera PolarScreen far-red fluorescence polarization assay format, which mitigates assay interference from autofluorescent compounds or scattered light through the use of a far-red tracer. The tracer shows substantially less interference from light scatter or autofluorescent library compounds than do fluorescein-based tracers, and gives rise to a larger assay window than the popular far-red fluorophore Cy5.     

The influence of glucocorticoid receptor single nucleotide polymorphisms on outcome after haematopoietic stem cell transplantation

Haematopoietic stem cell transplantation (HSCT) remains the only cure for most haematological malignancies, however, the mortality rate remains high. Complications after HSCT include relapse, graft versus host disease (GvHD), graft rejection and infection. Over the last few years several groups, have demonstrated that non‐HLA gene polymorphisms can be predictive of outcome after HSCT. Since the glucocorticoid cortisol is pivotal in the regulation of the immune system, we decided to examine single nucleotide polymorphisms (SNPs; rs6198, rs33388 and rs33389) within the glucocorticoid receptor (GR) and correlate with HSCT outcome. The training set consisted of patients (n = 458) who underwent HSCT for acute leukaemia between 1983 and 2005. In the recipients, the absence of the ACT haplotype and absence of the T allele of rs33388 were associated with decreased OS and the absence of the ACT haplotype, the absence of the T allele of rs33388 and the presence of the ATA haplotype were associated with increased risk of relapse. In addition, the presence of the ACT haplotype in the recipient showed a trend to be associated with increased risk of chronic graft versus host disease (cGvHD). The patients in this cohort received mainly myeloablative conditioning (n = 327). The SNPs in the glucocorticoid receptor were then investigated in a validation set (n = 251) of HSCT patients transplanted for acute leukaemia from 2006. This cohort contained significantly more patients that had received reduced intensity conditioning (RIC). Some of the results could be validated in these patients. However, contrary to the training set, the absence of the haplotype ACT in the donor in this cohort was associated with increased risk of cGvHD. Differences in the conditioning were shown to influence the results. These results are the first to associate GR SNPs with HSCT outcome and demonstrate the inherent problems of replicating SNP association studies in HSCT, due to different pre‐transplant regimens.     

A preliminary report of the dopamine receptor D4 and the dopamine transporter 1 gene polymorphism and its association with attention deficit hyperactivity disorder

Attention deficit hyperactivity disorder (ADHD) is one of the most prevalent childhood-onset psychiatric syndromes affecting 5%–10% of school-age children worldwide. Distortions in the catecholaminergic system seem to be responsible for this condition. Within this system there are several candidate genes, the dopamine receptor D₄ (DRD4) and the dopamine transporter 1 (DAT1), with common polymorphism which might be associated with ADHD. We performed a family based association study with 36 trios and 19 parent proband pairs. All diagnoses were confirmed by the “Hypescheme” diagnostic computer program. In this study we did not observe an association of ADHD with DRD4 and DAT1 polymorphism neither by the haplotype relative risk (HRR) method nor by the transmission disequilibrium test (TdT) method. The odds ratio for the DRD4 7-allele was 1.01 and 0.94 for both statistical tests, respectively, and the respective odds ratio for the DAT1 6-allele were 0.91 and 0.88.     

Test-Retest Reliability of a standardized psychiatric interview (DIS/CIDI)

The reliability of DSM-III diagnoses using an expanded version of the Diagnostic Interview Schedule (DIS), called the Composite International Diagnostic Interview (CIDI), was evaluated by examining 60 psychiatric inpatients on a test-retest basis. Acceptable agreement coefficients of (kappa) 0.5 or above were found for all but two disorders: dysthymic disorder and generalized anxiety disorder. The sub-classification of DSM-III affective disorders also revealed some discrepancies between the test and the retest interviews. When compared with results from earlier versions of the DIS, diagnostic reliability was found to have improved for the DSM-III anxiety disorders in particular. These improvements can possibly be attributed to some changes in the wording of the respective items of this section. Several reasons for lowered test-retest reliability are discussed.This research was supported by the German Research Foundation (DFG). The study was done in close collaboration with members of the Task Force on Instrument Development, established within the framework of the joint WHO/ADAMHA Project on Diagnosis and Classification. Lee Robins, PhD, John Helzer, MD, John Wing, MD, Norman Sartorius, MD, and Jack Burke, MD, provided us with helpful comments. Parts of the reliability analysis were done under contract with the NIMH, Division of Biometry and Epidemiology.     

Zur Darstellung von Pyrazino[3,2,1-j,k]carbazol-Derivaten

Synthesis of Pyrazino[3,2,1-j,k]carbazole Derivatives The synthesis of the tetrahydrocarbazolones 3a-g from the p-benzoquinones 6a/b and the 2-(aminomethylene)cyclopentanone derivatives 7 is reported as well as the cyclisation of the N-aminoacetonotrile derivative 3d to the title compound 8a.     

Ungesättigte Oxime, 27. Mitt. 5-Phenyl-2.4-pentadienale und ihre Oxime

Unsaturated Oximes, XXVII: 5-Phenyl-2,4-pentadienals and their Oximes The 5-phenyl-2.4-pentadienals 3a–f are synthesized from the 3-phenylpropenal diethyl acetals 1a–f and ethyl vinyl ether. Their oximes 4a–f can be separated into the (E)- and (Z)- isomers. With the α-branched aldehydes 5a–d , which were prepared by aldol condensation, only the (E)-oximes 6a–d are obtained.     

Rapid assay for the quantification of piretanide in biological fluids.

Piretanide was determined from plasma and urine using reversed-phase high-performance liquid chromatography. Plasma was extracted with diethylether at pH 4 using bumetanide as internal standard. The chromatographic separation was performed on an octadecylsilane column (ODS) with acetonitrile/pH 7 phosphate buffer (3:7, v/v). In order to determine piretanide in urine, bumetanide was added to a urine aliquot. Then the sample was centrifuged and the supernatant directly injected without extraction followed by chromatography on an ODS column with a mixture of methanol and pH 7 phosphate buffer (12:13, v/v). The compounds were detected by their intrinsic fluorescence, monitoring the eluate at 287/450 nm. Total chromatography times were 8 min for plasma and 13 min for urine samples. The method was applied for the assay of piretanide in plasma of healthy volunteers after i.v. or p.o. dosage of 6 mg piretanide.