Sensitive detection of platelet-specific antibodies with a modified MAIPA using biotinylated antibodies and streptavidin-coated beads.

We have developed a modified monoclonal antibody immobilization of platelet antigens assay (MAIPA) with enhanced sensitivity in detecting antibodies against human platelet antigens (HPA), using biotinylated monoclonal antibodies, streptavidin-coated beads and detection by flow cytometry. The beads-MAIPA gave superior signal-to-noise resolution (>10-fold higher) for detection of anti-HPA-1a and anti-HPA-5b compared with the in-house standard MAIPA. Also, efficient and reproducible detection of anti-HPA-15 (CD109) was shown. The enhanced sensitivity was confirmed using WHO International Reference Reagents for anti-HPA-1a, anti-HPA-3a and anti-HPA-5b, which allowed comparison of detection endpoints with other laboratories. Finally, the beads-MAIPA was validated for quantification of anti-HPA-1a. The lower limit of quantification was 0.4IU/mL for beads-MAIPA, compared to 1IU/mL previously reported for standard MAIPA. Based on improved performance against all HPA-antibodies tested, the beads-MAIPA has replaced the standard MAIPA in our laboratory in diagnostics of conditions due to HPA-immunization, such as fetal and neonatal alloimmune thrombocytopenia (FNAIT).

Determinants of long-term response to group therapy for dysfunctional fear of progression in chronic diseases.

Prior work demonstrated that cognitive-behavioral (CBT) and supportive-experiential (SET) group interventions can reduce dysfunctional fear of progression (FoP) in patients with chronic diseases. In this secondary analysis of a randomized controlled study, we investigated determinants of long-term response to group therapy for FoP. Response to therapy after 12 months was assessed using the Reliable Change Index (RCI). Outcome data were available for 129 patients with cancer and 116 patients with chronic arthritis. 37.9% of the patients in the CBT group and 32.7% of those attending the SET group indicated response to therapy (p=.402). Educational level predicted long-term response to therapy (OR 2.53, 95% CI 1.33-4.81; p=.005). Medical patients with lower education may need additional attention in order to gain long-lasting benefit from brief group psychotherapy. However, this investigation needs to be replicated in a study that includes a broader range of psychological predictors.

Higher rates of victimization to physical abuse by adults found among victims of school bullying.

Retrospective reports of exposure to physical abuse by an adult during childhood was assessed in 874 adolescents (426 boys, 448 girls; M age = 11.5 yr., SD = 0.8) who also reported whether they had been victimized by school bullying. Having been hit by an adult was significantly more common among victims of school bullying (39.5%) than among adolescents not victimized by school bullying (16.8%). No sex difference was found. The finding raises questions about whether victimization by physical abuse puts a child at greater risk for developing a "victim personality".

The immunophilin-like protein XAP2 is a negative regulator of estrogen signaling through interaction with estrogen receptor α.

XAP2 (also known as aryl hydrocarbon receptor interacting protein, AIP) is originally identified as a negative regulator of the hepatitis B virus X-associated protein. Recent studies have expanded the range of XAP2 client proteins to include the nuclear receptor family of transcription factors. In this study, we show that XAP2 is recruited to the promoter of ERα regulated genes like the breast cancer marker gene pS2 or GREB1 and negatively regulate the expression of these genes in MCF-7 cells. Interestingly, we show that XAP2 downregulates the E2-dependent transcriptional activation in an estrogen receptor (ER) isoform-specific manner: XAP2 inhibits ERα but not ERß-mediated transcription. Thus, knockdown of intracellular XAP2 levels leads to increased ERα activity. XAP2 proteins, carrying mutations in their primary structures, loose the ability of interacting with ERα and can no longer regulate ER target gene transcription. Taken together, this study shows that XAP2 exerts a negative effect on ERα transcriptional activity and may thus prevent ERα-dependent events.

[Fear of progression in chronic diseases]. / Progredienzangst bei chronischen Erkrankungen.

Chronically physically ill patients frequently suffer from psychological distress. The realistic fear of progression (FOP) of disease is one of the most important causes of distress in these patients. This study investigates the extent of FOP in patients with diverse diagnoses. 863 Patients answered the FOP-questionnaire, medical and sociodemographic items. Group differences were investigated with t- and F-tests. Determinants for FOP were identified by linear multiple regression. Some diagnostic subgroups differ in degree and profile of FOP. Patients with rheumatic diseases and Parkinson's disease score highest, patients with stroke or chronic peripheric vascular disease lowest in FOP total scores. Age, sex, economic situation and employment effect the level of fear of progression. Systematic studies are needed to verify the importance of FOP for patients with chronic disease and to ascertain clinical indication for psychotherapeutic support.

Cost-effectiveness of cognitive-behavioral group therapy for dysfunctional fear of progression in chronic arthritis patients.

BACKGROUND: Anxiety disorders are widespread in patients with chronic diseases such as rheumatoid arthritis (RA). This paper targets the cost-effectiveness analysis of a cognitive-behavioral group therapy (CBT) in comparison to a client-centered, supportive-experiential group therapy (SET) in arthritis patients with dysfunctional fear of progression. METHODS: From the societal perspective, direct costs were compared with the reduction of fear of progression over time. Means, their 95% confidence intervals (95% CI), the incremental cost-effectiveness graphic and the acceptability curve were obtained using 1000 non-parametric bootstrap replications. RESULTS: A total of 174 RA patients were included in the economic evaluation. The estimated means (95% CI) of direct costs and reduction of fear of progression were, respectively, €7945.34 (5075.59; 11335.08) and 0.25 (-0.48; 0.99) for patients in the SET and 5619.25 € (3950.67; 7708.52) and 0.94 (0.29; 1.62) for patients in the CBT. As the majority of the cost-effect pairs after bootstrap analysis were located in the southeast quadrant of the cost-effectiveness plane, the CBT can be considered a dominant intervention. CONCLUSION: The main result of our study is the higher cost-effectiveness of CBT in comparison to SET in RA patients with dysfunctional fear of progression.

Group psychotherapy of dysfunctional fear of progression in patients with chronic arthritis or cancer.

BACKGROUND: This study investigated the effectiveness of brief psychotherapeutic group interventions in reducing dysfunctional fear of disease progression (FoP). The interventions comprised either cognitive-behavioral group therapy or supportive-experiential group therapy. We tested whether these generic interventions would prove effective in different illness types. METHODS: Chronic arthritis in- patients (n = 174) and cancer in-patients (n = 174), respectively, were randomized to receive one of the two interventions. The patients provided data before intervention, at discharge, and at 3 and 12 months of follow-up. FoP was the primary outcome, secondary outcomes were anxiety, depression and quality of life. A treatment-as-usual control group provided data on the primary outcome. RESULTS: Patients with chronic arthritis indicated higher levels of FoP than cancer patients. The results revealed that, compared with no specialized intervention, both group therapies were effective in reducing dysfunctional FoP, but only among cancer patients. The effect sizes were 0.54 (cognitive-behavioral therapy) and 0.50 (supportive experiential therapy). The interventions were not differently effective in reducing the secondary outcomes. CONCLUSIONS: Dysfunctional FoP can be effectively targeted with brief group interventions. Psychotherapeutic interventions for reducing FoP should focus on specific illness characteristics.

Evaluation of two group therapies to reduce fear of progression in cancer patients.

PURPOSE: This paper aims to evaluate the effects of two psychotherapeutic interventions on dysfunctional fear of progression (FoP) in cancer patients and to investigate illness-specific influences. METHODS: One hundred seventy-four cancer patients were recruited from two rehabilitation clinics and randomly assigned to either a four-session cognitive-behavioral group therapy or a supportive-experiential group therapy. The main outcome criterion was FoP that was assessed with the Fear of Progression Questionnaire (FoP-Q) directly before (T1) and after (T2) the intervention, as well as 3 (T3) and 12 months (T4) after discharge. Secondary outcomes were anxiety, depression, and quality of life that were assessed with the following questionnaires: Questions on Life Satisfaction, Questionnaire for General Health Status, and the Hospital Anxiety and Depression Scale. Patients from the control group (n = 91) who received treatment as usual were recruited 1 year later with the same inclusion criteria and assessed with the FoP-Q at T1, T2, and T4. RESULTS: Analyses showed a significant main effect for time and a significant interaction for group x time for the main outcome variable. FoP decreased significantly over time in both intervention groups in contrast to the control group that showed only short-term improvements. The interventions were also effective in improving secondary outcomes except general life satisfaction. Analyses of cancer specific influences on FoP indicated a significant influence of disease status, i.e., patients with metastases and recurrence of cancer gained most from the interventions. CONCLUSIONS: Fear of progression, one of the main sources of distress for cancer patients, can be reduced with short psychotherapeutic interventions.

Fear of cancer progression and cancer-related intrusive cognitions in breast cancer survivors.

OBJECTIVE: To assess the character and frequency of fear of progression (FoP) and to clarify its relationship with cancer-related intrusive cognitions in breast cancer survivors. METHODS: A sample of 1083 patients was recruited in this cross-sectional study through a population-based Cancer Registry an average of 47 month following diagnosis (66% response rate). Participants completed self-report measures assessing fear of cancer progression (FoP-Q-SF), posttraumatic stress-disorder symptoms (PCL-C), coping strategies (DWI) and quality of life (QoL) (SF-8). RESULTS: In total, 23.6% of women were classified as having moderate to high FoP. Being nervous prior to doctors' appointments or examinations and being afraid of relying on strangers for activities of daily living were the most frequent fears. FoP was significantly associated with younger age, having children, disease progress, chemotherapy, perceived amount of impairments, physical and mental QoL, but not with time since initial diagnosis. Intrusive cognitions were screened in 37% of the sample. We found significant correlations between FoP and intrusive thoughts (r=0.63), avoidance (r=0.57), hyperarousal (r=0.54) and posttraumatic stress disorder diagnosis (r=0.42). Factors significantly associated with moderate and high FoP included a depressive coping style as well as an active problem-oriented coping style, intrusion, avoidance and hyperarousal symptoms (Nagelkerke's R(2)=0.44). CONCLUSIONS: Findings of this study give information regarding the frequency and the character of anxiety in breast cancer survivors and underline the relation of FoP to the reality of living with breast cancer. Results suggest that intrusive cognitions as well as avoidance and hyperarousal symptoms seem to be closely related to future-oriented fears of cancer recurrence.

Three QTLs for Botrytis cinerea resistance in tomato.

Tomato (Solanum lycopersicum) is susceptible to grey mold (Botrytis cinerea). Partial resistance to this fungus was identified in accessions of wild relatives of tomato such as S. habrochaites LYC4. In order to identify loci involved in quantitative resistance (QTLs) to B. cinerea, a population of 174 F(2) plants was made originating from a cross between S. lycopersicum cv. Moneymaker and S. habrochaites LYC4. The population was genotyped and tested for susceptibility to grey mold using a stem bioassay. Rbcq1, a QTL reducing lesion growth (LG) and Rbcq2, a QTL reducing disease incidence (DI) were identified. Rbcq1 is located on Chromosome 1 and explained 12% of the total phenotypic variation while Rbcq2 is located on Chromosome 2 and explained 15% of the total phenotypic variation. Both QTL effects were confirmed by assessing disease resistance in two BC(2)S(1) progenies segregating for either of the two QTLs. One additional QTL, Rbcq4 on Chromosome 4 reducing DI, was identified in one of the BC(2)S(1) progenies. F(2) individuals, homozygous for the Rbcq2 and Rbcq4 alleles of S. habrochaites showed a reduction of DI by 48%. QTLs from S. habrochaites LYC4 offer good perspectives for breeding B. cinerea resistant tomato cultivars.

[Fear of progression in breast cancer patients--validation of the short form of the Fear of Progression Questionnaire (FoP-Q-SF)]. / Progredienzangst bei Brustkrebspatientinnen--Validierung der Kurzform des Progredienzangstfragebogens PA-F-KF.

OBJECTIVES: Fear of progression is one of the most prevalent symptoms in cancer patients. The aim of this study was to validate the 12-item short version of the Fear of Progression Questionnaire (FoP-Q-SF). METHODS: A total of 1083 breast cancer patients were recruited by the Hamburg Cancer Register to fill out various questionnaires (response rate 67 %). RESULTS: Estimates of reliability were high (Cronbach's alpha = .87). The original one-factor structure was replicated. We used the HADS, the PCL-C, and the SF-8, among others, to validate the FoP-Q-SF. Significant positive correlations were found for fear of progression, anxiety and intrusion (r > .60) as well as for avoidance, hyperarousal and depression (r > or = .49). Moderate to high (negative) correlations were observed with health-related quality of life, in particular with the mental health dimensions (r > or = .48). Patients with cancer recurrence reported significant higher levels of fear of progression (p < .001). CONCLUSIONS: The short form of the Fear of Progression Questionnaire appears to be a reliable and valid instrument which can be recommended for further use in research and clinical care.

The co-chaperone XAP2 is required for activation of hypothalamic thyrotropin-releasing hormone transcription in vivo.

Transcriptional control of hypothalamic thyrotropin-releasing hormone (TRH) integrates central regulation of the hypothalamo-hypophyseal-thyroid axis and hence thyroid hormone (triiodothyronine (T(3))) homeostasis. The two beta thyroid hormone receptors, TRbeta1 and TRbeta2, contribute to T(3) feedback on TRH, with TRbeta1 having a more important role in the activation of TRH transcription. How TRbeta1 fulfils its role in activating TRH gene transcription is unknown. By using a yeast two-hybrid screening of a mouse hypothalamic complementary DNA library, we identified a novel partner for TRbeta1, hepatitis virus B X-associated protein 2 (XAP2), a protein first identified as a co-chaperone protein. TR-XAP2 interactions were TR isoform specific, being observed only with TRbeta1, and were enhanced by T(3) both in yeast and mammalian cells. Furthermore, small inhibitory RNA-mediated knockdown of XAP2 in vitro affected the stability of TRbeta1. In vivo, siXAP2 abrogated specifically TRbeta1-mediated (but not TRbeta2) activation of hypothalamic TRH transcription. This study provides the first in vivo demonstration of a regulatory, physiological role for XAP2.

Fear of progression in chronic diseases: psychometric properties of the Fear of Progression Questionnaire.

OBJECTIVE: The aim of this study was the development and psychometric testing of a new psychological questionnaire to measure the fear of progression (FoP) in chronically ill patients (cancer, diabetes mellitus and rheumatic diseases). METHODS: The Fear of Progression Questionnaire (FoP-Q) was developed in four phases: (1) generation of items (65 interviews); (2) reduction of items--the initial version of the questionnaire (87 items) was presented to 411 patients, to construct subscales and test the reliability; (3) testing the convergent and discriminative validity of the reduced test version (43 items) within a new sample (n=439); (4) translation--German to English. RESULTS: The scale comprised five factors (Cronbach's alpha >.70): affective reactions (13 items), partnership/family (7), occupation (7), loss of autonomy (7) and coping with anxiety (9). The test-retest reliability coefficients varied between .77 and .94. There was only a medium relationship to traditional anxiety scales. This is an indication of the independence of the FoP. Significant relationships between the FoP-Q and the patient's illness behaviour indicate discriminative validity. CONCLUSIONS: The FoP-Q is a new and unique questionnaire developed for the chronically ill. A major problem and source of stress for this patient group has been measuring both specifically and economically the FoP of an illness. The FoP-Q was designed to resolve this problem, fulfill this need and reduce this stress.

Evidence for a novel mechanism of time-resolved flavin fluorescence depolarization in glutathione reductase.

Time-resolved flavin fluorescence anisotropy studies on glutathione reductase (GR) have revealed a remarkable new phenomenon: wild-type GR displays a rapid process of fluorescence depolarization, that is absent in mutant enzymes lacking a nearby tyrosine residue that blocks the NADPH-binding cleft. Fluorescence lifetime data, however, have shown a more rigid active-site structure for wild-type GR than for the tyrosine mutants. These results suggest that the rapid depolarization in wild-type GR originates from an interaction with the flavin-shielding tyrosine, and not from restricted reorientational motion of the flavin. A novel mechanism of fluorescence depolarization is proposed that involves a transient charge-transfer complex between the tyrosine and the light-excited flavin, with a concomitant change in the direction of the emission dipole moment of the flavin. This interaction is likely to result from side-chain relaxation of the tyrosine in the minor fraction of enzyme molecules in which this residue is in an unsuitable position for immediate fluorescence quenching at the moment of excitation. Support for this mechanism is provided by binding studies with NADP+ and 2'P-5'ADP-ribose that can intercalate between the flavin and tyrosine and/or block the latter. Fluorescence depolarization analyses as a function of temperature and viscosity confirm the dynamic nature of the process. A comparison with fluorescence depolarization effects in a related flavoenzyme indicates that this mechanism of flavin fluorescence depolarization is more generally applicable.

Linkage disequilibrium mapping of yield and yield stability in modern spring barley cultivars.

Associations between markers and complex quantitative traits were investigated in a collection of 146 modern two-row spring barley cultivars, representing the current commercial germ plasm in Europe. Using 236 AFLP markers, associations between markers were found for markers as far apart as 10 cM. Subsequently, for the 146 cultivars the complex traits mean yield, adaptability (Finlay-Wilkinson slope), and stability (deviations from regression) were estimated from the analysis of variety trial data. Regression of those traits on individual marker data disclosed marker-trait associations for mean yield and yield stability. Support for identified associations was obtained from association profiles, i.e., from plots of P-values against chromosome positions. In addition, many of the associated markers were located in regions where earlier QTL were found for yield and yield components. To study the oligogenic genetic base of the traits in more detail, multiple linear regression of the traits on markers was carried out, using stepwise selection. By this procedure, 18-20 markers that accounted for 40-58% of the variation were selected. Our results indicate that association mapping approaches can be a viable alternative to classical QTL approaches based on crosses between inbred lines, especially for complex traits with costly measurements.

A physiologically representative in vitro model of the coronary circulation.

With the development of clinical diagnostic techniques to investigate the coronary circulation in conscious humans, the in vitro validation of such newly developed techniques is of major importance. The aim of this study was to develop an in vitro model that is able to mimic the coronary circulation in such a way that coronary pressure and flow signals under baseline as well as hyperaemic conditions are approximated as realistically as possible and are in accordance with recently gained insights into such signals in conscious man. In the present in vitro model the heart, the systemic and coronary circulation are modelled on the basis of the elements of a lumped parameter mathematical model only consisting of elements that can be represented by segments in an experimental set-up. A collapsible tube, collapsed by the ventricular pressure, represents the variable resistance and volume behaviour of the endocardial part of the myocardium. The pressure and flow signals obtained are similar to physiological human coronary pressure and flow, both for baseline and hyperaemic conditions. The model allows for in vitro evaluation of clinical diagnostic techniques.

Myocardial resistance assessed by guidewire-based pressure-temperature measurement: in vitro validation.

By injecting a few cubic centimeters of saline into the coronary artery and using thermodilution principles, mean transit time (T(mn)) of the injectate can be calculated and is inversely proportional to coronary blood flow. Because microvascular resistance equals distal coronary pressure (P(d)) divided by myocardial flow, the product P(d). T(mn) provides an index of myocardial resistance (IMR). In this in vitro study in a physiologic model of the coronary circulation, we compared IMR to true myocardial resistance (TMR) at different degrees of myocardial resistance and at different degrees of epicardial stenosis. Absolute blood flow was varied from 42 to 203 ml/min and TMR varied from 0.39 to 1.63 dynes. sec/cm(5). Inverse mean transit time correlated well to absolute blood flow (R(2) = 0.93). Furthermore, an excellent correlation was found between IMR and TMR (R(2) = 0.94). IMR was independent on the severity of epicardial stenosis and thus specific for myocardial resistance. Thus, using one single guidewire, both fractional flow reserve and IMR can be measured simultaneously as indexes of epicardial and microvascular disease, respectively, enabling separate assessment of both coronary arterial and microvascular disease.

Two parallel pathways mediate cytoplasmic localization of the dioxin (aryl hydrocarbon) receptor.

The dioxin receptor is a ligand-dependent transcription factor that mediates the biological effects of dioxin and related environmental pollutants. In the absence of ligand the receptor is present in the cytoplasmic compartment of the cell associated with the hsp90-dependent chaperone complex. This complex regulates several functions of the receptor such as ligand binding and nuclear import. Furthermore, intracellular localization of the receptor is modulated by multiple factors such as the export protein CRM-1 and the hsp90-associated immunophilin XAP-2. We have identified the mechanism of XAP-2-induced cytoplasmic localization of the receptor and studied the potential cross-talk between CRM-1 and XAP-2. We show that XAP-2 anchors the ligand-free receptor to cytoskeletal structures. This effect is blocked upon treatment with the actin inhibitor cytochalasin B, whereas the tubulin inhibitor colchicine had no effect on receptor localization. In addition, we show that the receptor interacts with CRM-1 both in the presence and absence of ligand. CRM-1-mediated nuclear export occurs independently of XAP-2. Our data provide evidence that CRM-1 and XAP-2 act in parallel through different mechanisms and target different interfaces of the receptor. These results suggest that two pathways cooperate to localize the non-activated receptor in the cytoplasmic compartment of the cell.

AFLPs mark different genomic regions compared with RFLPs: a case study in tomato.

A set of three tomato chromosome 7 introgression lines (ILs) containing overlapping segments of Lycopersicon pennellii DNA was screened with a set of 10 EcoRI-MseI and 10 PstI-MseI AFLP primer combinations. A large number of markers were identified that mapped to one of the four regions of chromosome 7 defined by the set of three ILs. Because many of the identified markers have known map positions in three tomato reference maps, their location on the tomato genome could be verified. It was demonstrated that the three chromosome 7 ILs carried a chromosome 10 region harbouring a cluster of six AFLP markers that had not been detected before using RFLPs. The causes and implications of this observation are discussed.