Basic characteristics and safety of donation in related and unrelated haematopoietic progenitor cell donors - first 10 years of prospective donor follow-up of Swiss donors.

Since July 2007 prospective life-long follow-up (FU) for unrelated (URD) and related donors (RD) is mandatory in Switzerland and data on every allogeneic haematopoietic progenitor cell (HPC) donation are collected prospectively. We report the real-world experience of HPC donation during a 10-year study period (01.07.2007-30.06.2017) with basic characteristics and FU data. 1105 donors underwent 1155 HPC donation procedures. Eighty percent of first donations performed by 802 (73%) RDs and 303 (27%) URDs were peripheral blood stem cells (PBSC), 20% bone marrow (BM). Male donors were over-represented as URD (60% male vs 40% female). Main differences between RDs and URDs concerned age and pre-existing health disorders. RDs were significantly older at first donation (median age 48 years) compared to URD (34 years, p < 0.0001) and had more pre-existing health problems: 25% vs 9% in URD (p < 0.0001). No fatal complications occurred, collection related severe adverse events (SAE) after first donation were not significantly different between groups (RD 1.2%, URD 0.99%), incidence rates for neoplastic and autoimmune diseases did not exceed the rates of the general population. RDs are a more heterogeneous and potentially more vulnerable group, but if donor evaluation is performed appropriately, HPC donation is still safe.

Prevotella strains and lactamic resistance gene distribution in different oral environments of children with pulp necrosis.

AIM: To determine the presence of Prevotella species, the cfxA/cfxA2, blaZ and blaTEM genes associated with resistance to lactamic agents in different oral niches of children with pulp necrosis. METHODOLOGY: Children with pulp necrosis in primary teeth had samples of saliva, supragingival, pulp chamber and root canal biofilms collected and tested for Prevotella species (P. intermedia, P. nigrescens, P. tannerae) and for beta-lactam resistance genes (cfxA/cfxA2, blaZ and blaTEM). The presence of bacterial DNA was examined through PCR, with a specific primer directed to the 16S rRNA gene. Specific primers were used to detect the Prevotella species and beta-lactam resistance genes. The chi-square test was used to analyse associations between the presence of bacteria and clinical variables. The Cochran's Q test was used to assess whether the proportion of gene detection is the same between different sites. RESULTS: Thirty-two teeth were sampled from 27 patients with a mean age of 5.5 years (±1.76). The total detection rate of Prevotella strains was 29.1%, 25%, 21.8% and 32.29% in saliva, supragingival, pulp chamber and root canal samples, respectively. P. nigrescens was the most commonly detected species in all oral niches. The previous use of antibiotics was associated with detection of P. nigrescens in saliva (P = 0.03). Pain was associated with the presence of P. nigrescens (P = 0.04) and P. tannerae (P = 0.01) in pulp chamber biofilm. blaTEM was detected in the four oral niches, being more frequent (23.8%) in supragingival biofilm (Cochran's Q test, P = 0.04). The presence of P. intermedia in SB and PC was associated with the detection of blaTEM in saliva (P = 0.04). The cfxA/cfxA2 and blaZ genes were not detected in any of the four oral niches. CONCLUSIONS: The oral cavity of children with pulp necrosis had a variable distribution of Prevotella strains in different niches. Saliva, supragingival biofilm, pulp chamber and root canals of primary teeth with necrotic pulps can harbour resistance genes to beta-lactams agents.

Common and well-documented HLA alleles over all of Europe and within European sub-regions: A catalogue from the European Federation for Immunogenetics.

BACKGROUND: A catalogue of common and well-documented (CWD) human leukocyte antigen (HLA), previously established by the American Society for Histocompatibility and Immunogenetics (ASHI), is widely used as indicator for typing ambiguities to be resolved in tissue transplantation or for checking the universality of any HLA allele in the world. However, European population samples, which are characterized by a substantial level of genetic variation, are underrepresented in the ASHI catalogue. Therefore, the Population Genetics Working Group of the European Federation for Immunogenetics (EFI) has facilitated data collection for an European CWD catalogue. MATERIALS AND METHODS: To this end, 2nd-field HLA-A, -B, -C,- DRB1,- DQA1,- DQB1 and -DPB1 data of 77 to 121 European population samples (21 571-3 966 984 individuals) from 3 large databases, HLA-net/Gene[VA], allelefrequencies.net and DKMS, were analysed. RESULTS: The total number of CWD alleles is similar in the EFI (N = 1048) and ASHI (N = 1031) catalogues, but the former counts less common (N = 236 vs 377) and more well-documented (N = 812 vs 654) alleles than the latter, possibly reflecting differences in sample numbers and sizes. Interestingly, approximately half of the CWD alleles reported by EFI were not reported by ASHI and vice-versa, underlining the distinct features of the two catalogues. Also, although 78 common alleles are widely distributed across Europe, some alleles are only common within specific sub-regions, showing regional variability. CONCLUSION: Although the definition of CWD alleles itself is affected by different parameters, calling for current updates of the list, the EFI CWD catalogue provides new insights into European population genetics and will be a very useful tool for tissue-typing laboratories in and beyond Europe.

The Use of Resin Composite Layering Technique to Mask Discolored Background: A CIELAB/CIEDE2000 Analysis.

The purpose of this study was to evaluate the ability of three resin composite systems to mask a severely discolored background by the application of a layering technique through CIELAB and CIEDE2000 analysis. Ninety 1.5-mm-thick disc specimens were produced from three different resin composite restoration systems: IPS Empress Direct (Ivoclar Vivadent), Charisma Diamond (Heraeus Kulzer), and Filtek Z350 XT (3M-ESPE). The specimens were divided into groups according to the restoration system and the resin composite shade combination used for the layering technique (enamel, body, and dentin shades). Color measurements were performed by a reflectance spectrophotometer (SP60, EX-Rite) against a C4 shade background and an inherent color background, which simulates a severely discolored background and a tooth surface with no discoloration, respectively. The total color difference between both color measurements was calculated by CIELAB (ΔE*ab) and CIEDE2000 (ΔE00) formulas. The mean ΔEab* and ΔE00 values were analyzed by analysis of variance (general linear models) and Tukey's post hoc tests (α=0.05). Three groups presented clinically acceptable color difference values (ΔE*≦3.46 and ΔE00≦2.25): 1.5 mm dentin, 1.0 mm dentin/0.5 mm body, and 1.0 mm dentin/0.5 mm enamel; ie, all the groups from the Z350 XT restoration system. The resin composite layering technique is an effective way to mask severely discolored backgrounds. The Filtek Z350 XT system was the only restoration system capable of masking the C4 background.

Color stability of a resin composite: Effect of the immersion method and surface treatments.

BACKGROUND: In vitro staining methods expose the entire specimen to staining solutions. In a real clinical situation, this is not observed, since one should consider that the bonded surface is not exposed to the oral environment. Theoretically, the clinical condition would be the best simulated if the specimens were exposed to staining solutions by partial immersion. AIMS: To evaluate if different immersion methods and surface treatments influence the color stability of resin-based specimens. METHODOLOGY: A stainless steel matrix was used to prepare 30 disc-shaped specimens that were randomly allocated in three groups: Without polishing, polishing with abrasive discs, and surface sealant. Half of the specimens were isolated to maintain only the upper surface exposed to staining (partial immersion) and the other half was totally immersed in coffee solution for 48 h (total immersion). The coordinates ΔE*, ΔL*, Δa*, Δb* were assessed by spectrophotometer. STATISTICAL ANALYSIS: Two-way ANOVA and Tukey's post hoc tests (α =0.05). RESULTS: Specimens submitted to partial immersion showed lower values of ΔE*, ΔL*, Δa*, Δb*, in comparison to total immersion (P = 0.000). Specimens covered by a surface sealant presented lower ΔE* values regardless of the immersion method. CONCLUSIONS: Specimens totally immersed in staining solutions could in somehow overestimate the color change, once that in most clinical conditions not all of the restoration surfaces are exposed to the oral environment. Moreover, as the surface sealant application produces color change values that are clinically acceptable, it might be used in esthetic restorations as an adjunct treatment.

16(th) IHIW: analysis of HLA population data, with updated results for 1996 to 2012 workshop data (AHPD project report).

We present here the results of the Analysis of HLA Population Data (AHPD) project of the 16th International HLA and Immunogenetics Workshop (16IHIW) held in Liverpool in May-June 2012. Thanks to the collaboration of 25 laboratories from 18 different countries, HLA genotypic data for 59 new population samples (either well-defined populations or donor registry samples) were gathered and 55 were analysed statistically following HLA-NET recommendations. The new data included, among others, large sets of well-defined populations from north-east Europe and West Asia, as well as many donor registry data from European countries. The Gene[rate] computer tools were combined to create a Gene[rate] computer pipeline to automatically (i) estimate allele frequencies by an expectation-maximization algorithm accommodating ambiguities, (ii) estimate heterozygosity, (iii) test for Hardy-Weinberg equilibrium (HWE), (iv) test for selective neutrality, (v) generate frequency graphs and summary statistics for each sample at each locus and (vi) plot multidimensional scaling (MDS) analyses comparing the new samples with previous IHIW data. Intrapopulation analyses show that HWE is rarely rejected, while neutrality tests often indicate a significant excess of heterozygotes compared with neutral expectations. The comparison of the 16IHIW AHPD data with data collected during previous workshops (12th-15th) shows that geography is an excellent predictor of HLA genetic differentiations for HLA-A, -B and -DRB1 loci but not for HLA-DQ, whose patterns are probably more influenced by natural selection. In Europe, HLA genetic variation clearly follows a north to south-east axis despite a low level of differentiation between European, North African and West Asian populations. Pacific populations are genetically close to Austronesian-speaking South-East Asian and Taiwanese populations, in agreement with current theories on the peopling of Oceania. Thanks to this project, HLA genetic variation is more clearly defined worldwide and better interpreted in relation to human peopling history and HLA molecular evolution.

The probability of identifying a 10/10 HLA allele-matched unrelated donor is highly predictable.

Identification of an unrelated HLA allele-matched hematopoietic stem cell (HSC) donor is a costly and time-consuming procedure. To improve search logistics, we have limited the search period to 6 months and have introduced a probability estimate of the chances of identifying a 10/10 HLA allele-matched donor. Probabilities were classified as high (>95%), intermediate (50%) and low (<5% chance) based on allele and haplotype frequencies. By analyzing 350 consecutive searches between 2002 and 2005 (1719 donors tested), the probability estimates turned out to be correct for 96% (high), 88% (low) and 56% (intermediate) patients. For searches with a high probability of success, at least one of the 10 most frequent haplotypes in Caucasoids was found in 69% of the patients, but in only 11% of the patients with a low-probability estimate (P<0.00001). Survival probability at 3 years was significantly higher for HSCT patients classified with a high-probability estimate when compared to patients in the intermediate/low-probability groups (74 vs 51 and 54% respectively, P=0.01). The same difference in survival probabilities was observed when only 10/10 matched unrelated HSCT patients were analyzed. In the intermediate-/low-probability groups, patients with alternative (haploidentical, autologous) or mismatched unrelated donors had similar survival estimates. Probability prediction is therefore feasible in the search process for unrelated donors and can guide the therapeutic strategy.

Toxicity of cadmium and copper on larval and juvenile stages of the estuarine crab Chasmagnathus granulata (Brachyura, Grapsidae).

The toxicity of cadmium and copper were assayed in the estuarine crab Chasmagnathus granulata. Acute assays were made on the first larvae stage, and both acute and chronic assays were made on juvenile crabs. The acute lethal toxicity of the assayed heavy metals was three orders of magnitude higher in larvae than in juveniles. Cadmium proved to be more toxic than copper in most cases; this difference was more evident in the chronic assays on juveniles, according to the nonphysiological feature of cadmium and its persistent accumulation in organisms. During these chronic assays, cadmium produced both a significant mortality and a clear retardation of molting, in accordance with its inhibitory effect on the molting process as previously reported. Copper only caused a molting acceleration during chronic assays, perhaps as a detoxifying mechanism. Heavy metal concentrations having effects in the chronic assays have been reported in some sectors of the estuary where the assayed species lives.

Epstein-Barr virus-dependent lymphoproliferative disease: critical role of IL-6.

Epstein-Barr virus (EBV)-induced lymphoproliferative disease (lpd) is a B cell neoplasm that affects patients who are immunosuppressed in the context of organ transplantation or HIV infection. A model for the aggressive form of this entity was generated by xenotransplantation of SCID mice with human peripheral blood leukocytes from individuals with prior contact with EBV. This model, where large B cell lymphoma occurs, was used to test the hypothesis that IL-6 has a major role in EBV-induced B cell tumorigenesis. IL-6 is known to differentiate B cells into immunoglobulin-secreting plasma cells and induce EBV replication, and xenochimeric animals have detectable serum levels of human IL-6. Human IL-6 inhibition with a neutralizing monoclonal antibody decreased tumor incidence from 62 % to 27 %. In addition, anti-IL-6 treatment significantly improved xenotransplanted animal survival, with median survival at > 245 days when compared to that of controls at 132 days. In conclusion, IL-6 plays a critical role in the pathogenesis of EBV-induced human lpd, and IL-6 inhibition may represent a new and promising preventive or therapeutic approach against this malignancy.

[The value of anticardiolipin antibody assessment]. / Intérêt du dosage des anticorps anticardiolipine.

Anticardiolipin antibodies (ACL) are associated with the presence of several clinical conditions. To determine the clinical interest of their evaluation, we reviewed all the results of ACL determinations performed in our laboratory during 1990 and 1991. 266 analyses (232 patients) were carried out. Determinations of IgG and IgM type ACL were performed by ELISA. ACL values were positive or slightly positive in 40% of the analyzed samples. Diagnosis was mentioned as the grounds for 212 requests (186 patients). ACL were positive (> m + 3SD) in about 40% of patients with autoimmune disease or with at least two of the following pathologies: autoimmune disease, venous or arterial thromboembolism, recurrent abortion.

Plasminogen activators in dextran sulfate-activated euglobulin fractions: a molecular analysis of factor XII- and prekallikrein-dependent fibrinolysis.

To elucidate the mechanism by which activation of the contact system of blood coagulation leads to expression of fibrinolytic activity, we have determined the molecular characteristics of the plasminogen activators present in dextran sulfate-treated euglobulin fractions by electrophoretic-zymographic analysis and specific immunoadsorption. In addition to free and protease inhibitor-bound tissue-type plasminogen activator (t-PA), dextran sulfate precipitates of euglobulins contained the complex formed between plasma kallikrein and C1-inhibitor, an indicator of prekallikrein activation. These precipitates also contained substantial fibrinolytic activity related to urinary-type plasminogen activator (u-PA). Autoradiographic analysis was then used to evaluate the cleavage of 125I-single-chain u-PA (prourokinase) in dextran sulfate euglobulins as well as after exposure to kallikrein or beta-factor XIIa. This analysis supported the conclusion that plasma kallikrein-mediated cleavage and activation of single-chain u-PA is the mechanism operative for the development of lytic activity in euglobulin precipitates following activation of the contact system.

Fibrinolysis in normal subjects--comparison between plasminogen activator inhibitor and other components of the fibrinolytic system.

We analyzed fibrinolytic parameters in 20 healthy men and 20 healthy women, aged from 25 to 59, before and after 10 and 20 min venous occlusion. The 10 min post-occlusion fibrinolytic activity measured directly in diluted unfractionated plasma by a highly sensitive 125I-fibrin plate assay correlated well with the activity of euglobulins determined by the classical fibrin plate assay (r = 0.729), but pre-stasis activities determined with these two methods did not correlate (r = 0.084). The enhancement of fibrinolytic activity after venous occlusion was mainly due to an increase of t-PA in the occluded vessels (4-fold increase t-PA antigen after 10 min and 8-fold after 20 min venous occlusion). Plasminogen activator inhibitor (PAI) activity and plasminogen activator inhibitor 1 (PAI-1)1 antigen levels at rest showed considerable dispersion ranging from 1.9 to 12.4 U/ml, respectively 6.9 to 77 ng/ml. A significant increase of PAI-1 antigen levels was observed after 10 and 20 min venous occlusion. At rest no correlation was found between PAI activity or PAI-1 antigen levels and the fibrinolytic activity measured by 125I-FPA. However, a high level of PAI-1 at rest was associated with a high prestasis antigen level of t-PA and a low fibrinolytic response after 10 min of venous stasis. Since the fibrinolytic response inversely correlated with PAI activity at rest, we conclude that its degree depends mainly on the presence of free PAI.

[Differentiation of a normal population and of a cohort of patients presenting with a history of idiopathic thromboembolism as identified with fibrinolytic tests]. / Discrimination d'une population normale et d'un collectif de patients ayant présenté un antécédent thromboembolique idiopathique au moyen des tests fibrinolytiques.

To identify persons at risk for development of thromboembolic disease, fibrinolytic and blood coagulation parameters of normal controls (n = 40) and patients (n = 40) have been compared. Four fibrinolytic parameters, i.e. fibrinolytic activity at rest (FAr), fibrinolytic response after 10 minutes of venous occlusion (delta FA10'), and resting antigenic levels of the plasmatic inhibitor of the plasminogen activators (PAI-1) and of urokinase (u-PA), showed significant differences between the two study groups. The separate and combined discriminant analysis of FAr and delta FA10' values revealed that individuals with FAr below 0.63 UI/ml or a delta FA10' below 0.98 UI/ml are at risk for development of thromboembolic disease.

Fibrinolysis in pregnancy: a study of plasminogen activator inhibitors.

During pregnancy the plasma concentration of two different inhibitors of plasminogen activators (PAIs) increases. The only one found in the plasma of nonpregnant women (PAI1) is immunologically related to a PAI of endothelial cells; its plasma activity, as deduced from the inhibition of single-chain tissue-type plasminogen activator (t-PA), increased from 3.4 +/- 2.3 U/mL (mean +/- 95% confidence limits) in the plasma of nonpregnant women to 29 +/- 7 U/mL at term, and its antigen level, measured by a radioimmunoassay, increased from 54 +/- 17 ng/mL to 144 +/- 25 ng/mL. In pregnancy plasma a second PAI (PAI 2) related to a PAI found in placenta extracts was observed. Its level, quantified with a radioimmunoassay, increased from below the detection limit (approximately 10 ng/mL) in normal plasma to 260 ng/mL at term. One hour after delivery, PAI 1 activities and antigen decreased sharply, but the PAI 2 antigen levels remained constant. Three days later, the PAI 1 antigen levels had fallen to normal levels, but the PAI 2 antigen levels were still at least eightfold above the nonpregnant values. During pregnancy, the t-PA and prourokinase (u-PA) antigen concentrations increased 50% and 200%, respectively, whereas the plasminogen and alpha 2-antiplasmin levels remained constant. Despite the large variations in the levels of PAs and PAIs, the overall fibrinolytic activity as measured in diluted plasma by a radioiodinated fibrin plate assay did not change significantly. Just after delivery, a great increase in the t-PA antigen levels was observed. Three to five days after delivery most parameters of the fibrinolytic system were normal again. Our results demonstrate that during pregnancy and in the puerperium profound alterations of the fibrinolytic system occur that are characterized by increases in PAs and their inhibitors, but these alterations do not affect the overall fibrinolytic activity.