Nationwide study of eculizumab in paroxysmal nocturnal hemoglobinuria: Evaluation of treatment indications and outcomes.

Eculizumab is an effective treatment for paroxysmal nocturnal hemoglobinuria (PNH). However, considering the risk of life-threatening meningococcal disease, life-long duration and costs, there are strict criteria for initiation of therapy. To evaluate the application and real-world effectiveness of eculizumab in the Netherlands, a multicenter retrospective cohort study was conducted: indications and treatment outcomes were collected for 105 Dutch PNH patients. In all patients, eculizumab was initiated conforming to indications as formulated in the Dutch PNH guideline. According to recently published response criteria, 23.4% of the patients had reached a complete hematological response, 53.2% a good or partial response, and 23.4% a minor response after 12 months of therapy. In the majority of patients the response remained stable during long-term follow-up. The degree and relevance of extravascular hemolysis significantly differed between response groups (p = 0.002). Improvements of EORTC-QLQc30 and FACIT-fatigue scores were observed, however patients reported lower scores than the general population. A detailed evaluation of 18 pregnancies during eculizumab showed no maternal or fetal deaths, and no thromboembolic events during pregnancy. This study demonstrates that the majority of patients benefit from eculizumab when adhering to the indications as formulated in the Dutch PNH guideline. However, novel therapies are needed to further improve real-world outcomes, such as hematological responses and quality of life.

OMIP - 081: A new 21-monoclonal antibody 10-color panel for diagnostic polychromatic immunophenotyping.

The 10-color panel consisting of 21 monoclonal antibodies (mAbs) is developed as a one-tube panel to detect leukemia and lymphoma cells in all hematopoietic cell lineages. In particular, this tube is mentioned for a fast screening to identify aberrant cells in samples suspected for malignant cell localization and to enable comprehensive immunophenotyping of samples with low cell counts. The panel contains mAbs for selection of the populations and mAbs against target antigens on the various hematopoietic maturation stages. Due to the limited number of PMTs in most used flow cytometers for clinical purposes, stacking of conjugates in one color is needed to include all relevant markers for simultaneous analysis of the aberrant cells. The 21-mAb panel is tested on peripheral blood (PB), and bone marrow (BM) samples and enables an efficient and correct identification of hematological malignancies. This panel improves the diagnostic potential.

Phase I/II Trial of a Combination of Anti-CD3/CD7 Immunotoxins for Steroid-Refractory Acute Graft-versus-Host Disease.

Effective therapies for treating patients with steroid-refractory acute graft-versus-host-disease (SR-aGVHD), particularly strategies that reduce the duration of immunosuppression following remission, are urgently needed. The investigated immunotoxin combination consists of a mixture of anti-CD3 and anti-CD7 antibodies separately conjugated to recombinant ricin A (CD3/CD7-IT), which induces in vivo depletion of T cells and natural killer (NK) cells and suppresses T cell receptor activation. We conducted a phase I/II trial to examine the safety and efficacy of CD3/CD7-IT in 20 patients with SR-aGVHD; 17 of these patients (85%) had severe SR-aGVHD, and all 20 patients had visceral organ involvement, including 18 (90%) with gastrointestinal (GI) involvement and 5 (25%) with liver involvement. A validated 2-biomarker algorithm classified the majority of patients (11 of 20) as high risk. On day 28 after the start of CD3/CD7-IT therapy, the overall response rate was 60% (12 of 20), with 10 patients (50%) achieving a complete response. The 6-month overall survival rate was 60% (12 of 20), including 64% (7 of 11) classified as high risk by biomarkers. The 1-week course of treatment with CD3/CD7-IT caused profound but transient depletion of T cells and NK cells, followed by rapid recovery of the immune system with a diverse TCR Vß repertoire, and preservation of Epstein-Barr virus- and cytomegalovirus-specific T cell clones. Furthermore, our results indicate that CD3/CD7-IT appeared to be safe and well tolerated, with a relatively low prevalence of manageable and reversible adverse events, primarily worsening of hypoalbuminemia, microangiopathy, and thrombocytopenia. These encouraging results suggest that CD3/CD7-IT may improve patient outcomes in patients with SR-aGVHD.

Bilateral Triple Talocalcaneal, Calcaneonavicular, and Talonavicular Tarsal Coalition: A Case Report.

We report a rare case of a female, aged 42 years, with symptomatic bilateral triple tarsal coalition, that is, talocalcaneal, calcaneonavicular, and talonavicular tarsal coalition. The patient was treated conservatively by adjusting her activities. At the 12-month follow-up, the patient was asymptomatic. Bilateral triple tarsal coalition is a rare disorder, especially in nonsyndromic patients. The purpose of this case report was to highlight this rare type of multiple bilateral tarsal coalitions and to discuss the relevant existing literature.

Immunophenotypic analysis of erythroid dysplasia in myelodysplastic syndromes. A report from the IMDSFlow working group.

Current recommendations for diagnosing myelodysplastic syndromes endorse flow cytometry as an informative tool. Most flow cytometry protocols focus on the analysis of progenitor cells and the evaluation of the maturing myelomonocytic lineage. However, one of the most frequently observed features of myelodysplastic syndromes is anemia, which may be associated with dyserythropoiesis. Therefore, analysis of changes in flow cytometry features of nucleated erythroid cells may complement current flow cytometry tools. The multicenter study within the IMDSFlow Working Group, reported herein, focused on defining flow cytometry parameters that enable discrimination of dyserythropoiesis associated with myelodysplastic syndromes from non-clonal cytopenias. Data from a learning cohort were compared between myelodysplasia and controls, and results were validated in a separate cohort. The learning cohort comprised 245 myelodysplasia cases, 290 pathological, and 142 normal controls; the validation cohort comprised 129 myelodysplasia cases, 153 pathological, and 49 normal controls. Multivariate logistic regression analysis performed in the learning cohort revealed that analysis of expression of CD36 and CD71 (expressed as coefficient of variation), in combination with CD71 fluorescence intensity and the percentage of CD117+ erythroid progenitors provided the best discrimination between myelodysplastic syndromes and non-clonal cytopenias (specificity 90%; 95% confidence interval: 84-94%). The high specificity of this marker set was confirmed in the validation cohort (92%; 95% confidence interval: 86-97%). This erythroid flow cytometry marker combination may improve the evaluation of cytopenic cases with suspected myelodysplasia, particularly when combined with flow cytometry assessment of the myelomonocytic lineage.

Short-Term Clinical Outcome of Hemiarthroplasty Versus Arthrodesis for End-Stage Hallux Rigidus.

Few data are available to compare the outcomes of first metatarsophalangeal joint (MTPJ) hemiarthroplasty and arthrodesis. We included 46 patients who had undergone BioPro(®) first MTPJ hemiarthroplasty and 132 who had undergone arthrodesis, with a minimum follow-up duration of 12 months. The primary outcome was patient satisfaction, which was determined using binominal questions. The Foot and Ankle Outcome Score, Foot Function Index, and Numerical Rating Scale for pain and limitations questionnaires were also used. The secondary outcome was treatment failure. No differences were found in the satisfaction rate (p = .54) after a median period of 38.4 (range 12 to 96) months and 39.8 (range 12 to 96) months in the hemiarthroplasty and arthrodesis patients, respectively. Furthermore, no differences were found in the failure rates (p = .93) or the interval to failure (p = .32).The results of the present study showed no significant differences in the short-term clinical outcomes and failure rates for BioPro(®) first MTPJ hemiarthroplasty and arthrodesis. Prospective comparative studies are required to determine whether BioPro(®) first MTPJ hemiarthroplasty is a good alternative for first MTPJ arthrodesis in the long term.

Demonstration of atomic force microscopy imaging using an integrated opto-electro-mechanical transducer.

The low throughput of atomic force microscopy (AFM) is the main drawback in its large-scale deployment in industrial metrology. A promising solution would be based on the parallelization of the scanning probe system, allowing acquisition of the image by an array of probes operating simultaneously. A key step for reaching this goal relies on the miniaturization and integration of the sensing mechanism. Here, we demonstrate AFM imaging employing an on-chip displacement sensor, based on a photonic crystal cavity, combined with an integrated photodetector and coupled to an on-chip waveguide. This fully-integrated sensor allows high-sensitivity and high-resolution in a very small footprint and its readout is compatible with current commercial AFM systems.

Joint preservation surgery for correction of flexible pes cavovarus in adults.

BACKGROUND: Cavovarus foot deformity often results from muscular imbalance. If deformities are still flexible, surgical correction can be aimed primarily at improving muscular balance. MATERIALS AND METHODS: We retrospectively assessed the outcome of 19 procedures (in 15 patients) to evaluate patient satisfaction. All patients had a flexible deformity. The surgical procedures that were used included soft-tissue releases, tendon transfers and lengthenings, and osteotomies of either the first metatarsal or the calcaneus. The surgical results in terms of pain, function and alignment of the foot were evaluated by means of standardized questionnaire (AOFAS foot score and the Dutch version of the Foot Function Index), patient interview, physical examination and X-ray. The changes in calcaneal pitch and talus-metatarsal I angle were calculated. RESULTS: Overall patient satisfaction was good. The results were very satisfactory in five procedures, satisfactory in ten, moderately satisfactory in two, unsatisfactory in one, and very unsatisfactory in one procedure. The AOFAS hindfoot score was on average 82.5 +/- 16. The talus-metatarsal I angle significantly decreased from 22.5 to 17 degrees (p = 0.002). The anatomical corrections of the foot did not show association with the patients' clinical characteristics. CONCLUSION: Our results show that in joint preserving flexible pes cavovarus correction patient satisfaction is generally good. Radiographic alignment of the foot was not significantly associated with patient-based outcome in this small series of patients.

Lateral column lengthening for acquired adult flatfoot deformity caused by posterior tibial tendon dysfunction stage II: a retrospective comparison of calcaneus osteotomy with calcaneocuboid distraction arthrodesis.

In this study, clinical and radiological results after lateral column lengthening by calcaneocuboid distraction arthrodesis and calcaneus osteotomy were compared. Thirty-three patients (35 feet) treated with lateral column lengthening by distraction arthrodesis (14 patients, 16 feet; group I) or by calcaneus osteotomy (19 patients, 19 feet; group II) for adult-acquired flatfoot deformity caused by stage II posterior tibial tendon dysfunction were compared retrospectively. Mean follow-up was 42.4 months (range, 6-78 months) for group I and 15.8 months (range, 6-32 months) for group II (P < .001). The American Orthopaedic Foot & Ankle Society ankle-hindfoot score was determined, 4 variables were measured on preoperative and postoperative weight-bearing radiographs, and a number of independent and outcome variables, including patient satisfaction, were recorded. Group 2 had a significantly higher American Orthopaedic Foot & Ankle Society score compared with group I (mean, 85 vs. 72, respectively; P < .02) at time of last follow-up, and there were no dissatisfied patients in group I, whereas 2 patients in group II were dissatisfied with the result of the operation. All radiological results were significantly better at time of follow-up in both groups (except for talocalcaneal angle in group I), although no significant differences were noted in the amount of change in radiographic measurements between the groups. No significant correlation was found between follow-up time and radiographic improvement, indicating stable radiographic measurements over time. In group II, 13 mild calcaneocuboid subluxations were observed. In both groups, 1 nonunion and 1 wound complication occurred. Based on our experience with the patients described in this report, we recommend lateral column lengthening by means of calcaneus osteotomy rather than distraction arthrodesis of the calcaneocuboid joint, for correction of stage II posterior tibial tendon dysfunction.

Mechanical and Electric Control of Photonic Modes in Random Dielectrics.

Random dielectrics defines a class of non-absorbing materials where the index of refraction is randomly arranged in space. Whenever the transport mean free path is sufficiently small, light can be confined in modes with very small volume. Random photonic modes have been investigated for their basic physical insights, such as Anderson localization, and recently several applications have been envisioned in the field of renewable energies, telecommunications, and quantum electrodynamics. An advantage for optoelectronics and quantum source integration offered by random systems is their high density of photonic modes, which span a large range of spectral resonances and spatial distributions, thus increasing the probability to match randomly distributed emitters. Conversely, the main disadvantage is the lack of deterministic engineering of one or more of the many random photonic modes achieved. This issue is solved by demonstrating the capability to electrically and mechanically control the random modes at telecom wavelengths in a 2D double membrane system. Very large and reversible mode tuning (up to 50 nm), both toward shorter or longer wavelength, is obtained for random modes with modal volumes of the order of few tens of (λ/n)3 .

Outcome after 52 Salto Ankle Prostheses Implanted by a Single Surgeon.

While ankle arthrodesis was traditionally the gold standard method of treatment for disabling end-stage ankle arthritis, total ankle replacement (TAR) has been an acceptable alternative. The satisfaction rate of patients with TAR however differs. The purpose of our study is to investigate whether implant survival and results with special emphasis on the satisfaction rate of patients treated with a TAR implanted by a single surgeon were comparable to the literature. This was a retrospective cohort study in a teaching hospital. Data was collected from 52 patients who received a total ankle replacement (TAR) between 05/2002 and 06/2014. The mean follow-up time was 4.2 years (95% CI 3.3 - 5.0). Results showed a high satisfaction rate of 94% and 94% survival of the TAR after 5 years. We conclude that TAR with the Salto prosthesis is, in our hands, a reliable solution for end-stage ankle arthritis, with results comparable to the literature.

Flow cytometric CD34+ stem cell enumeration: lessons from nine years' external quality assessment within the Benelux countries.

BACKGROUND: A biannual external quality assurance (EQA) scheme for flow cytometric CD34+ haematopoietic stem cell enumeration has been operational in the Benelux countries since 1996. In an evaluation of the results of 16 send-outs, we studied the effects of the methods used on assay outcome and whether or not this exercise was effective in reducing between-laboratory variation. METHODS: Data were analyzed using robust multivariate regression. This approach is relatively insensitive to outliers and is used to assess the effect of methodological aspects of CD34+ cell counting on the bias and variability. RESULTS: Five variables were associated with significant bias of absolute CD34+ cell counts: (i) unique laboratory number (ULN), (ii) gating strategy; (iii) CD34 mAb fluorochrome; (iv) type of flow cytometer, and (v) method of sample preparation. In addition, ULN and platform methodology (i.e., single vs. dual) contributed significantly to the variability of this assay. Overall, the variability in results of CD34+ cell enumeration has declined with time; in particular, after a practical workshop in which participants were trained to use the "single platform ISHAGE protocol." CONCLUSIONS: Between-laboratory variation in CD34+ cell enumeration can be reduced by standardization of methodologies between centres. Our approach, i.e., EQA with targeted training and feedback in response to reported results, has been successful in reducing the variability of CD34+ cell enumeration between participants.

Monocytes and macrophages in flow: an ESCCA initiative on advanced analyses of monocyte lineage using flow cytometry.

In April 2013, a symposium was organized to highlight different aspects of differentiation and activation of the monocyte-macrophage lineage as analyzed on the flow cytometer. Characterization of this lineage requires knowledge of the maturation process from their progenitors that are present in bone marrow up to the mature monocytic cells in peripheral blood, because each monocytic lineage cell with an aberrant phenotype refers to the corresponding maturation stage. A standardized quantitative analysis will facilitate the monitoring of the pathological processes and the clinical features, such as the outcome of treatment. However, changes in marker expression by variation in intensity, asynchronism, and lineage infidelity must be considered. The dynamics of normal marker expressions in early differentiation stages, e.g. molecules like HLA II, CD64 or CD14, give rise to a hypothesis on their possible role in monocyte ontogeny. Besides their usual role in tissue homeostasis, mature macrophages may also play a similar role in hematopoiesis. This meeting highlighted the large potential of flow cytometric tools available for monitoring of all these aspects in the monocytic and macrophage cell lineage. © 2015 International Clinical Cytometry Society.

Integrated nano-opto-electro-mechanical sensor for spectrometry and nanometrology.

Spectrometry is widely used for the characterization of materials, tissues, and gases, and the need for size and cost scaling is driving the development of mini and microspectrometers. While nanophotonic devices provide narrowband filtering that can be used for spectrometry, their practical application has been hampered by the difficulty of integrating tuning and read-out structures. Here, a nano-opto-electro-mechanical system is presented where the three functionalities of transduction, actuation, and detection are integrated, resulting in a high-resolution spectrometer with a micrometer-scale footprint. The system consists of an electromechanically tunable double-membrane photonic crystal cavity with an integrated quantum dot photodiode. Using this structure, we demonstrate a resonance modulation spectroscopy technique that provides subpicometer wavelength resolution. We show its application in the measurement of narrow gas absorption lines and in the interrogation of fiber Bragg gratings. We also explore its operation as displacement-to-photocurrent transducer, demonstrating optomechanical displacement sensing with integrated photocurrent read-out.

Fifteen years of external quality assessment in leukemia/lymphoma immunophenotyping in The Netherlands and Belgium: A way forward.

In 1985, external quality assurance was initiated in the Netherlands to reduce the between-laboratory variability of leukemia/lymphoma immunophenotyping and to improve diagnostic conclusions. This program consisted of regular distributions of test samples followed by biannual plenary participant meetings in which results were presented and discussed. A scoring system was developed in which the quality of results was rated by systematically reviewing the pre-analytical, analytical, and post-analytical assay stages using three scores, i.e., correct (A), minor fault (B), and major fault (C). Here, we report on 90 consecutive samples distributed to 40-61 participating laboratories between 1998 and 2012. Most samples contained >20% aberrant cells, mainly selected from mature lymphoid malignancies (B or T cell) and acute leukemias (myeloid or lymphoblastic). In 2002, minimally required monoclonal antibody (mAb) panels were introduced, whilst methodological guidelines for all three assay stages were implemented. Retrospectively, we divided the study into subsequent periods of 4 ("initial"), 4 ("learning"), and 7 years ("consolidation") to detect "learning effects." Uni- and multivariate models showed that analytical performance declined since 2002, but that post-analytical performance improved during the entire period. These results emphasized the need to improve technical aspects of the assay, and reflected improved interpretational skills of the participants. A strong effect of participant affiliation in all three assay stages was observed: laboratories in academic and large peripheral hospitals performed significantly better than those in small hospitals. © 2015 International Clinical Cytometry Society.

Relationship of plasma lipids to renal function and length of time on maintenance hemodialysis.

Significant hypertriglyceridemia, the most common lipid abnormality in renal failure, first occurs when the creatinine clearance falls to 50 ml/min. The prevalence of hypertriglyceridemia continues to rise as creatinine clearance falls further with the highest rate developing at a creatinine clearance less than 10 ml/min. Hypertriglyceridemia is correlated with plasma glucagon levels but not growth hormone or insulin. Plasma cholesterol values remain normal in the face of deteriorating renal function and show no correlation with any of the hormones measured. Although all three hormones became elevated as renal function diminished, none were directly correlated with glomerular filtration rate. There was a distinct decrease in the prevalence of hyperlipidemia after 5 years of maintenance hemodialysis therapy. Plasma growth hormone and glucagon through an effect on plasma triglyceride and plasma insulin by effecting plasma cholesterol may play a role in this decline of hyperlipidemia with duration of hemodialysis.