Risk factors for RhD immunisation in a high coverage prevention programme of antenatal and postnatal RhIg: a nationwide cohort study.

OBJECTIVE: To evaluate which risk factors for RhD immunisation remain, despite adequate routine antenatal and postnatal RhIg prophylaxis (1000 IU RhIg) and additional administration of RhIg. The second objective was assessment of the current prevalence of RhD immunisations. DESIGN: Prospective cohort study. SETTING: The Netherlands. POPULATION: Two-year nationwide cohort of alloimmunised RhD-negative women. METHODS: RhD-negative women in their first RhD immunised pregnancy were included for risk factor analysis. We compared risk factors for RhD immunisation, occurring either in the previous non-immunised pregnancy or in the index pregnancy, with national population data derived from the Dutch perinatal registration (Perined). RESULTS: In the 2-year cohort, data from 193 women were eligible for analysis. Significant risk factors in women previously experiencing a pregnancy of an RhD-positive child (n = 113) were: caesarean section (CS) (OR 1.7, 95% CI 1.1-2.6), perinatal death (OR 3.5, 95% CI 1.1-10.9), gestational age >42 weeks (OR 6.1, 95% CI 2.2-16.6), postnatal bleeding (>1000 ml) (OR 2.0, 95% CI 1.1-3.6), manual removal of the placenta (MRP) (OR 4.3, 95% CI 2.0-9.3); these factors often occurred in combination. The miscarriage rate was significantly higher than in the Dutch population (35% versus 12.-5%, P < 0.001). CONCLUSION: Complicated deliveries, including cases of major bleeding and surgical interventions (CS, MRP), must be recognised as a risk factor, requiring estimation of fetomaternal haemorrhage volume and adjustment of RhIg dosing. The higher miscarriage rate suggests that existing RhIg protocols need adjustment or better compliance. TWEETABLE ABSTRACT: Complicated delivery (caesarean section, manual removal placenta, major bleeding) is the most valid risk factor for RhD immunization despite antenatal and postnatal RhIg.

Neurocognitive functioning of children with mild to borderline intellectual disabilities and psychiatric disorders: profile characteristics and predictors of behavioural problems.

BACKGROUND: The aim of the current study was twofold: first, to uncover a neurocognitive profile of normative and relative strengths and weaknesses that characterises an extremely vulnerable group of children with mild to borderline intellectual disabilities (MBID) and co-morbid psychiatric disorders, and second, to investigate the relevance of these neurocognitive functions explaining internalising and externalising symptoms. METHOD: We recruited 45 children (Mage  = 9.5, SDage  = 1.7; range 6-13 years) with MBID (Full-Scale IQ 50-85) and at least one psychiatric disorder. Neurocognitive functioning was examined utilising the Wechsler Intelligence Scale for Children - Fifth Edition (WISC-V) indices and the Cognitive Task Application (COTAPP), a comprehensive computerised self-paced task designed in such a manner that 'g' (an overall tendency of children with MBID to execute tasks with a slower reaction time and a higher error rate) has been corrected for in the administration of the task (i.e. completely self-paced) and in the operationalisation of outcome measures. Behavioural problems were measured using the CBCL and TRF. One-sample t-tests and binomial tests were carried out to compare performance with normative data. Regression analyses were used to examine the relationship between neurocognitive parameters and mental health. RESULTS: Compared with normative data, very small to very large effect sizes were found, indicating clear heterogeneity amongst neurocognitive domains relevant for children with MBID. Two prominent neurocognitive weaknesses emerged: processing speed - characterised by slowness and unstableness combined with a high drift rate and delayed processing of the previous trial, particularly under higher cognitive demands - and working memory - in terms of a weaker central executive and 'slave' systems to temporarily store information. Both domains were not clearly predictive of internalising or externalising problems. CONCLUSION: Children with MBID and psychiatric disorders are hampered by a strongly diminished processing speed and working memory capacity, together resulting in an overall limited processing capacity that may underlie the general developmental delays on domains that depend on fast and parallel processing of information (i.e. language, reading, mathematics and more complex forms of social cognition). Neurocognitive vulnerabilities are neither necessary nor sufficient to explain internalising and externalising problems; rather, a mismatch between the support needs and adaptations these children need, arising from their diminished processing capacity, and the inadequacy of the environment to compensate for this vulnerability may be of relevance.

International Society of Blood Transfusion Working Party on red cell immunogenetics and blood group terminology: Cancun report (2012).

The International Society of Blood Transfusion Working Party on red cell immunogenetics and blood group terminology convened during the International congress in Cancun, July 2012. This report details the newly identified antigens in existing blood group systems and presents three new blood group systems.

Evaluation of candidate control genes for diagnosis and residual disease detection in leukemic patients using 'real-time' quantitative reverse-transcriptase polymerase chain reaction (RQ-PCR) - a Europe against cancer program.

Real-time quantitative RT-PCR (RQ-PCR) is a sensitive tool to monitor minimal residual disease (MRD) in leukemic patients through the amplification of a fusion gene (FG) transcript. In order to correct variations in RNA quality and quantity and to calculate the sensitivity of each measurement, a control gene (CG) transcript should be amplified in parallel to the FG transcript. To identify suitable CGs, a study group within the Europe Against Cancer (EAC) program initially focused on 14 potential CGs using a standardized RQ-PCR protocol. Based on the absence of pseudogenes and the level and stability of the CG expression, three genes were finally selected: Abelson (ABL), beta-2-microglobulin (B2M), and beta-glucuronidase (GUS). A multicenter prospective study on normal (n=126) and diagnostic leukemic (n=184) samples processed the same day has established reference values for the CG expression. A multicenter retrospective study on over 250 acute and chronic leukemia samples obtained at diagnosis and with an identified FG transcript confirmed that the three CGs had a stable expression in the different types of samples. However, only ABL gene transcript expression did not differ significantly between normal and leukemic samples at diagnosis. We therefore propose to use the ABL gene as CG for RQ-PCR-based diagnosis and MRD detection in leukemic patients. Overall, these data are not only eligible for quantification of fusion gene transcripts, but also for the quantification of aberrantly expressed genes.

CD antigens 2001.

This paper reviews the Seventh Human Leucocyte Differentiation Antigen (HLDA7) workshop. Due to the limitations of "blind" antibody screening, which had been evident at the previous meeting in 1996, participants at HLDA7 adopted a more selective approach to the choice of antibodies by identifying new CD specificities. This resulted in the addition of more than 80 new CD specificities. Plans for the eighth and subsequent workshops are also previewed.

SDF-1-induced actin polymerization and migration in human hematopoietic progenitor cells.

OBJECTIVE: The capacity of hematopoietic progenitor cells (HPCs; CD34(+) cells) to respond to chemotactic stimulation is essential for their homing efficiency, e.g., during stem cell transplantation. Previous studies established that stromal cell-derived factor-1 (SDF-1) and its receptor CXCR-4 play an important role in the homing of HPCs. The aim of the present study was to analyze SDF-1-induced actin polymerization and migration of HL-60 cells and primary human CD34(+) cells. MATERIALS AND METHODS: SDF-1-induced migration of CD34(+) cells from cord blood (CB) and peripheral blood (PB) across fibronectin-coated filters was measured in a Transwell assay. Actin polymerization was detected using fluorescent phalloidin and analyzed by confocal microscopy and FACS analysis. RESULTS: SDF-1 induced a rapid and transient increase in actin polymerization and in polarization of the actin cytoskeleton in primary CD34(+) cells and HL-60 cells. SDF-1 was found to induce significantly more actin polymerization in CB CD34(+) cells that show fast migration in vitro compared to slow migrating PB CD34(+) cells. Moreover, CB CD34(+) cells that had migrated toward SDF-1 showed an elevated and prolonged rise in F-actin upon second exposure to SDF-1 compared to nonmigrated cells, although both cell types expressed equal levels of the SDF-1 receptor CXCR-4. CONCLUSIONS: The relatively high migratory capacity of CB-derived human HPCs is not related to cellular polarization or high expression of the SDF-1 receptor but is largely determined by their capacity to efficiently polymerize F-actin in response to SDF-1.

Clinical relevance of third permanent molars in relation to crowding after orthodontic treatment.

OBJECTIVES: The purpose of this study was to determine the relationship between dental crowding and the clinical presence or absence of third permanent molar teeth. METHODS: Ninety-nine patients were analysed before and after orthodontic treatment and at least three years after the end of retention. The sample consisted of four groups: subjects whose third permanent molar teeth had erupted into the mouth, were non-erupted, were extracted, and were congenitally absent. Arch Length Discrepancy, Irregularity Index and observer bias were examined. Multivariate analysis of variance with repeated measurements was used to analyse differences between the four groups. RESULTS: Significant differences in Arch Length Discrepancy during time were shown between the premolar segment and the frontal area. The group with third permanent molar teeth congenitally missing showed a significant higher positive Arch Length Discrepancy in the premolar segment of the upper jaw. No significant differences in Irregularity Index were found between the third molar groups. CONCLUSIONS: It can be concluded that there is no relation between crowding and the presence or absence of third permanent molar teeth.

Peripheral blood stem cell (PBSC) mobilization and transplantation (PSCT) in patients with malignant lymphomas and solid tumors.

Peripheral blood stem cells can reconstitute bone marrow function after high-dose chemo-/radiotherapy. We describe 44 patients related with a three-day course of chemotherapy, for hematopoietic stem cell mobilization, consisting of cyclophosphamide or ifosfamide and etoposide (malignant lymphoma and germ cell tumor) or a one-day course of 5-fluorouracil, epirubicin and cyclophosphamide (breast cancer), followed by the administration of recombinant human granulocyte colony-stimulating factor (G-CSF). Maximum numbers peripheral blood stem cells (PBSC) were recruited on day 9-10 of the G-CSF administration. The total number of PBSC cells harvested with median 3.6 leukaphereses was 46 x 10(4)/kg (7.5-136) CFU-GM or 8 x 10(6)/kg (0.7-25.0)CD34+ cells for patients with solid tumors and 26 (4.5-258) CFU-GM's or 6.1 (1-0-39.2) CD34+ cells for patients with malignant lymphomas. Thirty-five patients with malignant lymphomas or solid tumors received high-dose chemotherapy followed by bone marrow and PBSC infusion (n = 8) or PBSC cell infusion alone (n = 27). The recovery of granulocytes, platelets and reticulocytes after peripheral stem cell transplantation (-PSCT) in addition to or instead of bone marrow, was markedly accelerated compared with the infusion of BM alone. The accelerated haemopoietic recovery was associated with a reduction in platelet and red blood cell transfusion, reduction in fever periods and earlier discharge from hospital. PSCT is an important alternative to autologous bone marrow transplantation (ABMT). This transplantation technique may also allows application of multiple-cycle intensive chemotherapy.

High-risk childhood acute lymphoblastic leukemia in first remission treated with novel intensive chemotherapy and allogeneic transplantation.

Children with acute lymphoblastic leukemia (ALL) and high minimal residual disease (MRD) levels after initial chemotherapy have a poor clinical outcome. In this prospective, single arm, Phase 2 trial, 111 Dutch and Australian children aged 1-18 years with newly diagnosed, t(9;22)-negative ALL, were identified among 1041 consecutively enrolled patients as high risk (HR) based on clinical features or high MRD. The HR cohort received the AIEOP-BFM (Associazione Italiana di Ematologia ed Oncologia Pediatrica (Italy)-Berlin-Frankfurt-Münster ALL Study Group) 2000 ALL Protocol I, then three novel HR chemotherapy blocks, followed by allogeneic transplant or chemotherapy. Of the 111 HR patients, 91 began HR treatment blocks, while 79 completed the protocol. There were 3 remission failures, 12 relapses, 7 toxic deaths in remission and 10 patients who changed protocol due to toxicity or clinician/parent preference. For the 111 HR patients, 5-year event-free survival (EFS) was 66.8% (±5.5) and overall survival (OS) was 75.6% (±4.3). The 30 patients treated as HR solely on the basis of high MRD levels had a 5-year EFS of 63% (±9.4%). All patients experienced grade 3 or 4 toxicities during HR block therapy. Although cure rates were improved compared with previous studies, high treatment toxicity suggested that novel agents are needed to achieve further improvement.

The impact of social support upon intention to leave among female nurses in Europe: secondary analysis of data from the NEXT survey.

OBJECTIVES: This study examined the importance of one's social work environment in the light of prevention of premature leave from the nursing profession. A research model with social support (from direct supervisor and close colleagues) as predictor and intention to leave as the dependent variable has been tested, while controlling for job satisfaction and age. Moreover, we have studied the impact of nurses' age upon the prevalence of social support from both parties. PARTICIPANTS: Data were obtained from 17,524 registered female nurses working in hospitals throughout Europe (Belgium, Germany, Finland, France, Italy, The Netherlands, Poland, and Slovakia). RESULTS: Our findings indicated that a lack of job satisfaction is an important risk factor in the light of nurses' turnover as for most countries the intention to leave cannot be buffered by social support from one's close colleagues. However, in general, social support from one's direct superior appeared to contribute negatively to the intention to leave the profession, over and above job satisfaction and age. As regards age effects, in line with our expectation, we have found a significant negative relationship between age and social support from close colleagues, while the hypothesis regarding the relationship between age and supervisory support could not be confirmed. CONCLUSIONS: Given its importance in the light of preventing premature leave, we advocate not to neglect the possible positive effects of social support from important key figures like nurses' direct supervisor and close colleagues. It is necessary for health care institutions to carefully pay attention to finding opportunities to obtain more social support for all staff members. In Section 5, limitations and practical implications of this study are dealt with.

Hepatitis B virus infection in an institution for the mentally retarded.

A cross-sectional survey on the prevalence of hepatitis B serologic markers and hepatitis B virus DNA was performed in a population of 493 mentally handicapped males. Special interest was focused on age-related variables such as age at entry into the institution and on duration of residency. Furthermore, the differences with regard to the prevalence of hepatitis B markers found in Down's syndrome residents and other mentally retarded persons were analyzed. In a longitudinal study, the impact of the presence of hepatitis B virus DNA in serum was studied. Overall, 62.1 per cent of residents had serologic evidence of infection with hepatitis B virus, while 16.7 per cent of those residents with markers of infection were positive for hepatitis B surface antigen (HBsAg). Hepatitis B virus DNA was found in 24 per cent of HBsAg carriers (all positive for hepatitis B e antigen (HBeAg). In residents whose age at entry was less than 15 years, those with Down's syndrome were more often carriers of HBsAg than other mentally retarded residents. In addition, Down's syndrome residents more often had serum hepatitis B virus DNA compared with residents with other forms of mental retardation. A young age at entry was recognized as an important factor with regard to the prevalence of hepatitis B markers. From the longitudinal studies, it appeared that loss of hepatitis B virus DNA from serum indicated imminent loss of HBeAg and normalization of alanine aminotransferase values. Knowledge of the hepatitis B virus DNA status of HBsAg carriers in these institutions may therefore provide a valuable tool in attempts to reduce the transmission of this infection.

Monoclonal antibody JSB-1 detects a highly conserved epitope on the P-glycoprotein associated with multi-drug-resistance.

Resistance to multiple chemotherapeutic agents is a common clinical problem in the treatment of cancer. This resistance may occur before primary therapy or be acquired during treatment. We have generated a monoclonal antibody (MAb) (JSB-I), specific for a conserved epitope on the plasma membrane 170- to 180-kDa glycoprotein, the expression of which is strongly correlated with the degree of multi-drug resistance (MDR). JSB-I strongly binds to both Chinese-hamster-derived MDR cell lines and human MDR cell lines, including cell lines derived from lung and ovary. A drug-sensitive revertant line, and the corresponding drug-sensitive parent lines, showed only weak reactivity or none at all. JSB-I reacts strongly to air-dried or acetone-fixed cells and therefore has potential value for diagnostic detection of MDR cells in human tumor samples.

Consensus strategy to quantitate malignant cells in myeloma patients is validated in a multicenter study. Belgium-Dutch Hematology-Oncology Group.

Recently the Belgium-Dutch Hematology-Oncology group initiated a multicenter study to evaluate whether myeloma patients treated with intensive chemotherapy benefit from additional peripheral stem cell transplantation. To determine treatment response accurately, we decided to quantitate malignant cells. To test a consensus quantitation strategy, 5 centers independently determined the immunoglobulin heavy chain sequences of patient tumor cells and developed allele-specific oligonucleotides (ASO) and ASO-polymerase chain reaction (PCR). We compared the reproducibility of real-time quantitation with quantitation using limiting dilutions. We distributed DNA samples with a 4-log range of tumor cell concentrations and found average quantitation values deviating 74% and 42% from the input values with real-time PCR (1 center) and limiting dilutions (4 centers), respectively. Within single centers we found an average variation coefficient of 0.74, with limiting dilutions not significantly different from the average 0.82 center-to-center variation coefficient. Within a single center, real-time quantitation proved more reproducible (average variation coefficient, 0.36). Quantification was confirmed in 3 patients during treatment in the protocol. This report shows that real-time PCR or limiting dilution assays can be used for quantitation in a single multicenter trial. We present a consensus strategy that allows an accurate comparison of quantitation data generated in independent centers.

CD antigens 2001.

The most recent Human Leucocyte Differentiation Antigen Workshop ("HLDA7") took place in 2000 in Harrogate, UK and the proceedings are about to be published (Leucocyte Typing VII). New Sections were introduced in this Workship (Dendritic cells, Stem/progenitor cells, Erythroid cells and Carbohydrate Structures) and monoclonal antibodies were selected for which at least some molecular data were already available (to avoid "blind" screening of reagents against known specificities). A total of more than 80 new CD specificities were established (previously the average was less than 30 new CD specificities per Workshop) and these are listed in this article. There is already evidence for the existence of many new leucocyte surface molecules for study at the next HLDA Workshop (in Adelaide in 2004), and we have listed in this article a number of such potential CD candidates (identified following the production of monoclonal antibodies or via gene cloning). There are also today an increasing number of lineage- and/or stage-restricted leucocyte-associated molecules localised within the cell cytoplasm (or nucleus): they will certainly prove of intense in the future for many laboratories studying human haematopoietic cells (regardless of whether a new "intracellular CD" categorisation scheme is devised for such molecules).