The purposes of this study were to present a prototype of a bracket-positioning gauge, which makes vertical inclination of the instrument difficult, allowing a reduction of vertical bracket ...positioning error, and to test its accuracy in bracket positioning by groups of individuals with different clinical experience and in specific groups of teeth.
For the testing of the prototype, four groups of six participants each were used: Group 1 was composed of undergraduate students in the dental school, who had no previous experience in bonding orthodontic attachments; Group 2 was composed of orthodontic graduate students in the dental school; Group 3 consisted of orthodontists with a maximum of 5 years of clinical experience; Group 4 comprised orthodontists with more than 5 years of clinical experience. A typodont was simulated with a Class I crowded malocclusion, which reproduced the same occlusal characteristics for all groups to be bonded. All participants were instructed to bond 0.022×0.028-in Edgewise brackets on the labial surfaces of the upper and lower incisors, canines, and premolars at a height of 4 mm from the incisal edge or the labial cusp tip.
Only the mean value of Group 1 showed statistically significant difference in the comparison with the standard measurement. In the groups of teeth, the difference was significant for the premolar and incisor groups.
Clinical experience interfered with the accuracy of vertical positioning of orthodontic attachments. As for the groups of teeth, premolars, followed by canines and incisors had the closest mean values to the standard measurement.
Background:The isolation of cell-free DNA (cfDNA) evolved the concept of liquid biopsy. Several works have analyzed the presence of cfDNA in lymphomas, especially in diffuse large B-cell lymphoma ...(DLBCL) and Hodgkin lymphoma (HL), however there is limited information regarding the detection of cfDNA in other types of lymphomas or the role of cfDNA detection to monitor disease outcome.
Objectives:1.- To analyze the presence of cfDNA at diagnosis of patients with lymphoproliferative malignancies. 2.- To evaluate the change in concentration of cfDNA (cfDNA) after treatment in DLBCL patients.
Material and Methods:A retrospective study in a single center was performed including 221 adult patients since January 2015 to February 2019 with: DLBCL, HL, marginal zone lymphoma (MZL), follicular lymphoma (FL), chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), lymphoplasmacytic lymphoma (LPL) and peripheral T-cell lymphomas (TCL). All patients had plasma samples collected at diagnosis that were stored in the institutional biobank (MarBiobank). The cfDNA were obtained from 1 mL of -80°C frozen stored plasma using the MagMax Cell Free DNA isolation kit (Thermo Fisher Scientific). The cfDNA quantification was performed using the Qubit system with the dsDNA high sensitivitykit (Thermo Fisher) and is expressed as ng/mL of plasma. In addition, patients with DLBCL who were treated with rituximab-CHOP/CHOP-like regimens were evaluated for cfDNA analysis, pre and post-treatment, and results were compared with PET-CT scan findings.
Results: Stored plasma samples at diagnosis were available in 221 patients: DLBCL 82, HL 22, CLL/LPL 13, FL 36, MZL 37, MCL: 11, TCL 10 and others 10 (B-cell lymphoma unclassified, hairy cell leukemia, Burkitt lymphoma). Successful identification of cfDNA was obtained in 95.9% (212/221) of samples. DLBCL patients showed higher cfDNA globally and, DLBCL, HL and FL patients showed a higher concentration than MZL who exhibited the lower concentration of all groups (p=0.009; p=0.013 and p=0.002); see table 1.
50 DLBCL patients with a median follow-up since diagnosis of 25.5 (5-51) months were analyzed. Median age was 67 (19-79) years, males 56% (28). IPI distribution: low-risk 28% (14), low-intermediate 26% (13), high-intermediate 24% (12) and high risk 22% (11) cases. Detection of cfDNA was successful in 100% at diagnosis and in 98% (49) cases post-therapy. The mean cfDNA at diagnosis was 2.21 (standard deviation- SD: 1.60) ng/mL with a correlation with LDH concentration (p<0.001) and with high-risk IPI category (p=0.02). The mean cfDNA in the post-therapy sample was 4.39 (SD 16.46) ng/mL.
After therapy 86% (43) of patients achieved at least PR (41 complete response), and the mean cfDNA for these patients was 1.58 (SD 1.96); patients who showed no response or progressive disease after therapy exhibited higher cfDNA 21.25 ng/mL (SD 41.91)(p<0.001). In order to evaluate the clinical relevance of the changes of cfDNA after treatment, we considered a variation of +/-25% of cfDNA at diagnosis regarding cfDNA after therapy to classify the results. Therefore, 13 patients had an increase of cfDNA after therapy, 6 patients had no change, and 31 patients had a decrease. Patients with a decrease in cfDNA at the end of therapy were less likely to relapse (p<0.01) During follow-up, two patients relapsed after 24 and 8 months. The patient who relapsed after 24 months showed an increase in cfDNA, from 0.966 ng/mL to 20.80ng/mL three months before the histological confirmation of relapse.
Conclusions:Isolation of cfDNA was feasible in >95% of lymphoma patients independently of histology or disease stage. Patients with DLBCL exhibited the higher cfDNA concentration which were also correlated with LDH concentrations and high-risk IPI. Kinetics of cfDNA is related to response to therapy in DLBCL and also might detect relapse. Even though additional studies are necessary, monitoring of cfDNA may help in management of patients with DLBCL.
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Salar:Roche: Research Funding, Speakers Bureau; Janssen Pharmaceuticals: Consultancy, Speakers Bureau; Gilead: Consultancy, Speakers Bureau; Celgene: Consultancy. Sanchez-Gonzalez:Takeda: Consultancy, Speakers Bureau; Alexion: Speakers Bureau; Gilead: Speakers Bureau; Shire: Speakers Bureau; Novartis: Consultancy, Speakers Bureau; Amgen: Consultancy, Speakers Bureau. Gimeno:JANSSEN: Consultancy, Speakers Bureau; Abbvie: Speakers Bureau. Bellosillo:Qiagen: Consultancy, Speakers Bureau; TermoFisher Scientific: Consultancy, Speakers Bureau.
INTRODUCTION
The diagnosis of CMML according to WHO 2017 requires the presence of ≥1x109/L and ≥10% of monocytes in peripheral blood (PB). Establishing an accurate diagnostic is difficult since many ...clinical situations present persistent monocytosis. The presence of dysplasia is frequent but not always present and cytogenetic aberrations are infrequent in this disease (20-25% of cases). Although 85-90% of CMML patients present ≥1 mutation in TET2, SRSF2 or ASXL1, the use of NGS panels is not widespread. The study of PB monocyte subsets by flow cytometry (FC) has gained interest for CMML diagnosis. The increase of classical monocytes (Mo1) upper 94% presents a high sensitivity (Sn) and specificity (Sp) for CMML diagnosis (Sn 90.6, Sp 95.1; Selimoglu-Buet et al, Blood 2015). The 94% threshold was validated in two studies (Talati C et al, Blood 2017; Tarfi S et al, Blood Cancer J 2018). However, some controversies have recently appeared in the literature. Picot T detected the 95% cutoff as the one with the best Sn (100%) and Sp (97%) (Picot T et al, Front Oncol 2018). Hudson CA found that the presence of < 1.13% (Sn 100, Sp 96) of non-classical monocytes (Mo3) was the best predictor for CMML diagnosis (Hudson CA et al, Am J Clin Pathol 2018). With the exception of the study of Tarfi S, based on 47 CMML, the rest presented a very low number of patients (Talati C: 29; Picot T: 15; Hudson CA: 16) and therefore a bias could be expected specially when studying the Sn of the proposed methods. Moreover, the different series assessing the “monocyte assay” have no molecular data and therefore this could diminish the accuracy of the results since some patients may have received misdiagnoses.
The aim of our study was to assess the Sn and Sp of different thresholds of Mo1 and Mo3 in a large series with well-annotated clinical, cytogenetic and molecular data. Moreover, we assessed whether the study of CD2 and CD56 monocyte expression in combination with the %Mo1 >94 test improves the detection of the disease.
METHODS
50 CMML, 12 MDS, 11 MPN with ≥1x109/L monocytes and 79 reactive monocytosis with ≥1x109/L monocytes (N = 152) were prospectively studied from 02/2016 to 07/2019. We studied PB monocyte subsets by FC: Mo1 (CD14bright/CD16-), Mo2 (CD14bright/CD16+) and Mo3 (CD14dim or -/CD16bright). In addition, we assessed the expression of CD56 and CD2 in monocytes (positivity ≥ 20%). Finally, targeted NGS of the entire exonic sequence of 25 genes recurrently mutated in myeloid malignancies was performed (VAF sensitivity: 2%). Chi-Square or Fisher exact tests were used as appropriate. ROC curves were developed to explore optimal cutoffs in terms of sensitivity (Sn) and specificity (Sp). Moreover, we plotted the AUC of the subset of Mo1 and Mo3. Finally, the Youden index (YI) was used to detect the threshold of Mo1 and Mo3 with the best balance between Sn and Sp.
RESULTS AND DISCUSSION•The Sn and Sp of the Mo1>94% test in our series were similar to those reported by the French group (GFM). Our Sn and Sp were 90% and 92% respectively with a YI of 82. The Sn and Sp of the Mo1>93% were 94% and 84% with a YI of 78. Finally, the 95% cutoff proposed by Picot T et al showed a Sn of 81% and a Sp of 96% with a YI of 77. Therefore, the 94% cutoff presented the best balance between Sn and SP of the different thresholds assessed.•The Mo3 threshold of 1.13% proposed by Hudson CA et al showed a Sn of 67% and a Sp of 95% with a YI of 62. The best Mo3 cutoff in our series was established in 3.18% with a Sn of 90% and Sp of 83%. The YI of this threshold was 73.•The AUC for the percentage (%) of Mo1 (0.937, IC 95%: 0.89-0.99) was better than the AUC of the % of Mo3 (0.924, IC 95%: 0.88-0.97) reinforcing the use of %Mo1 as the item with the best discriminative power for CMML diagnosis. The AUC of the percentage of Mo1 population was similar to that reported by the GFM (Figure 1).•The Sn and Sp for CD56 expression in monocytes was 67% and 91% respectively, while CD2 expression showed a Sn of 38% and a Sp of 99%.•Finally, the presence of at least one of the following: Mo1 >94%, CD56+ or CD2+ presented the highest Sn (98%) and a Sp of 84%. This method may be a very good screening test due to the low false negative rate expected. This combined approach showed the best balance between Sn and Sp (YI: 82).
CONCLUSIONS
Our study supports the utility of the Mo1 >94% test as the best flow cytometry assay for establishing accurate diagnoses in CMML. The combined assay of Mo1, CD56 and CD2 may be of high utility as a screening test.
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Bellosillo:Qiagen: Consultancy, Speakers Bureau; TermoFisher Scientific: Consultancy, Speakers Bureau.
Introduction:
MYC rearrangements (MYCr) occur in 5 to 15% of diffuse large B-cell lymphomas (DLBCL) and 20 to 35% of high-grade B-cell lymphoma, NOS (HGBL-NOS), are a defining criterion of the ...category HGBL with rearrangements of MYC and/or BCL2/BCL6 (HGBL, with MYCr and BCL2/BCL6), and may be present in 90% of Burkitt lymphoma. The current WHO classification considers cytogenetic techniques as the appropriate tool to detect MYCr but does not define how to approach to the identification of such alteration. As the global incidence of MYCr in large B-cell lymphomas (LBCL) is low, it is necessary to clarity whether FISH or other cytogenetic methods have to be applied to all LBCL or only in selected cases. We previously identified LMO2 as a potential surrogate marker of MYCr in LBCL (Colomo L, Am J Surg Pathol 2017). Our aim with this study is to confirm this observation and evaluate the clinical impact of this marker in the survival of patients with LBCL.
Methods:
We have prospectively studied between September 2014 and July 2019 a new series of 180 LBCL including patients with DLBCL, HGBL, with HGBL, with MYCr and BCL2/BCL6, HGBL-NOS and transformed low-grade lymphomas into DLBCL (tDLBCL) diagnosed according to WHO criteria. LMO2 (clone 1A9-1), MYC (clone Y69) and a common immunohistochemistry (IHC) panel of B and T-cell markers have been used for the histological categorization of the cases, using whole tissue sections. The cutoff for LMO2 and MYC were 30% and 40%, respectively. MYC and BCL6 genes were studied using break apart probes, and BCL2 gene using dual-color dual-fusion probes (IGH/BCL2), all from Vysis-Abbott. We have statistically correlated the loss of expression of LMO2 and the overexpression of MYC with the presence or absence of MYCr. Moreover, we performed survival analyses assessing the clinical impact of LMO2 in a series of 162 LBCL patients (112 DLBCL, 20 HGBL, with MYCr and BCL2/BCL6, 4 HGBL-NOS and 26 tDLBCL). The survival series included cases diagnosed before 2014 with IHC and FISH data.
Results:
The prospective series included 132 patients with DLBCL (78M/52F; median age 67 years, range 35-95), 9 HGBL, with MYCr and BCL2/BCL6 (5M/4F; median age 67 years, range 42-85), 4 HGBL-NOS (2M/2F; median age 58 years, range 42-89), and 35 tDLBCL (31 transformed follicular lymphomas, 3 marginal zone lymphoma and 1 lymphoplasmacytic lymphoma; 23M/20F; median age 64 years, range 40-82).
LMO2 and MYC were expressed as follows, respectively: 84/130 (65%) and 46/132 (35%) in DLBCL; 1/9 (11%) and 8/9 (89%) in HGBL, with MYCr and BCL2/BCL6; 0/4 and 3/4 (75%) HGBL-NOS; 25/34 (73%) and 7/33 (21%) tDLBCL. MYCr were identified in 9/132 (7%) DLBCL; all HGBL, with MYCr and BCL2/BCL6; 4/4 HGBL-NOS; 7/35 (20%) tDLBCL.
The table shows the comparisons between LMO2 and MYC protein expression for the identification of the presence of MYCr in the series of LBCL. Whereas in the whole series LMO2 and MYC had similar results, among CD10-positive cases, LMO2 had better results than MYC and identified better the presence of MYCr than MYC protein expression.
The 5-year progression-free survival (PFS) according the diagnostic categories was 59% for DLBCL, 28% for HGBL, with MYCr and BCL2/BCL6, 25% for HGBL-NOS and 22% for tDLBCL (P=0.015). In addition, PFS was significantly lower for the presence of MYCr (26% vs 53%, P=0.02) and MYC IHC expression (35% vs 53%, P=0.005), and showed a positive trend for LMO2 loss of expression (39% vs 52%, P=0.1). The 5-year overall survival (OS) according the diagnostic categories was 67% for DLBCL, 23% for HGBL, with MYCr and BCL2/BCL6, 50% for HGBL-NOS and 77% for tDLBCL (P<0.001). In addition, OS was significantly shorter for the presence of MYCr (37% vs 71%, P=0.002), MYC protein expression (46% vs 75%, P=0.001), and for LMO2 loss of expression (46% vs 74%, P=0.005). In a Cox regression survival analysis including IPI and LMO2 for the 68 CD10-positive cases, IPI (HR: 1.61 P=0.03) was the most important variable for predicting OS, and LMO2 showed a significant trend (HR: 0.44 P=0.06). However, the addition of MYC IHC and MYCr did not add predictive accuracy to IPI score (HR: 1.6 P=0.31; HR: 1.8 P=0.19, respectively).
Conclusions:
LMO2 detection by IHC is a useful tool to detect MYCr in aggressive LBCL, particularly in CD10-positive cases. Moreover, LMO2 protein expression captures the prognostic significance of the different diagnostic histological categories and the presence of MYCr in this group of lymphomas.
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Sanchez-Gonzalez:Alexion: Consultancy, Honoraria; Gilead: Consultancy, Honoraria; Shire: Consultancy, Honoraria; Amgen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Takeda: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Novartis: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees. Salar:Celgene: Consultancy; Gilead: Consultancy, Speakers Bureau; Janssen: Consultancy, Speakers Bureau.
The management of IgM monoclonal gammopathies undetermined significance (IgM-MGUS) and Waldenstrom's macroglobulinemia (WM) may be challenging. Modern immunoassays that quantify specific monoclonal ...heavy and light chain immunoglobulins are promising for their use in these applications.
Ninety consecutive patients (39 IgM-MGUS, 32 indolent WM iWM, and 19 WM) seen between January 2007 and March 2014 were analyzed. Heavy/light chain (HLC) and serum free light chains assays (FLC) were determined at diagnosis to study their utility as biomarkers in IgM monoclonal gammopathies.
The HLC involved to uninvolved IgM ratios (iHLC/uHLC) showed a progressive increase when going from IgM-MGUS, to iWM and to WM (p=0.002). Furthermore, an iHLC/uHLC>62 identified a group of iWM patients with a shorter time-to-progression (TTP) (108 vs. 133 months, p=0.033). Separate analysis of the involved and uninvolved components showed that only the suppression of the uninvolvedimmunoglobulin was predictive of shorter TTP (HR=3.04, p=0.03) suggesting that it could be the majorcontributor to the prognostic value of the Hevylite assay. Additionally, a multivariate analysis showed that immunosuppression (either classical immunoparesis or Hevylite immunosuppression) was an independent prognostic factor (p=0.016) reinforcing its relevance in the disease mechanism. Finally, monoclonal sFLC levels were highest in WM patients, with 83% presenting values>60 mg/L.
The results suggest that the levels of immunosuppression and/or the iHLC/uHLC ratio of IgM immunoglobulins measured by Hevylite are associated with greater disease activity which significantly impacts in the outcome of WM patients and may also help in the differentiation of IgMMGUS from iWM.
Introduction: Patients with type1 Gaucher's disease (GD1) have an increased risk of gammopathy (RR,33 Taddei TH 2009), multiple myeloma (RR,25.), other haematological malignancies (RR,3.45) and ...overall cancer risk (RR, 1.80). The Spanish Registry of Gaucher Disease (SpRGD) was established in 1993 in response to the need to group individual experiences in the diagnosis and management of this disease, increasing knowledge related to general characteristics and to know the real incidence and prevalence in the Spanish population. Registration is open to all physicians involved in the management of patients with GD and offers free enzymatic analysis, biomarkers and molecular analysis for the diagnosis and monitoring of patients (www.feeteg.org).
Aim: to analyses the incidence of malignancies in adults GD patients.
Patients and methods: A review of the SpRGD to obtain data form patients over 20 years of age at May, 2016 was performed. Physicians on charge fulfilled a survey in which they inform about the incidence of malignancies and follow-up information. Ethical approval was obtained from the institutional board and all patients has signed an inform consent before to be included into the SpRGD.
Results: Of the 281 adult patients (³20 years) included, 279 were GD1 and 2 GD3. The average age of the entire cohort was 52.3 (23-90), of which 140 men, 141 women. Of these, 27 (9.6%) patients with GD1, 5 homozygous for N370S and 22 heterozygous for N370S had the presence of a malignancy and / or monoclonal gammopathy (MGUS), two of them had more than one neoplasia. Male / female: 11/16, mean age 60.2 (25-90), median follow-up of 16.5 years (4-23). Six have died by the tumor complications. All MGUS (N=12) were identified at GD diagnosis, they were 6 males and 6 females mean age 55.5 y (10-82) of them 50% under 60 years of age. Sixteen patients developed seventeen different neoplasms, with a female predominant (11, 68.7%). Only eight patients were under therapy at the time of neoplasia diagnosis (table1). Mean time on therapy 7.4 years (1.2-13-6). Neoplasms were registered (M/ F): B cell malignancies: Hodgkin lymphoma 1 (M), chronic lymphocytic leukemia 1 (M), multiple myeloma 1 (M), myeloid neoplasms: chronic myeloid leukemia: 1 (F), myelodysplastic syndrome: 1 (F), solid tumors: melanoma: 1 (F), meningioma: 2 (F), uterine cancer: 3 (F), gastric carcinoma 1 (F), cancer colon 2 (F), breast cancer 1 (F), prostate adenocarcinoma: 1(M), lung cancer 1 (M), liver carcinoma 1 (M), thyroid cancer 1 (F).
Conclusions: It has been widely reported the highest incidence of haematological malignancies among patients with GD. Nevertheless in this cohort of Spanish patients, the incidence of solid tumors is similar to haematological neoplasms in general and higher than B cell lymphoid. Probably the incidence of malignancy in this population and during this monitoring period is similar to the expected in Spanish general population found in 0.21% / year, however females showed two times risk increase for malignancies and this aspect warranty further studies.
This work has been carried out with aid for research FIS PS15/00616 and FEETEG
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No relevant conflicts of interest to declare.
MYC
rearrangements (
MYC
-R) confer unfavorable prognosis to large B-cell lymphomas (LBCL). Because of the low incidence of such genetic alteration, surrogates to screen
MYC
-R may be useful in daily ...practice. Previous studies suggested that clone 1A9-1 of LMO2 loss may be a good predictor for the presence of
MYC
-R in LBCL. The present study examines the utility of LMO2 clone SP51. For this purpose, we have analyzed 20 Burkitt lymphomas and 325 LBCL. Among them, 245 cases were studied prospectively using whole tissue sections, and 100 retrospectively by tissue microarrays. The cohort of CD10-positive prospective cases achieved the best results. Lack of LMO2 SP51 expression predicted the presence of
MYC
-R with high specificity, accuracy, positive and negative predictive value (PPV/NPV), and positive and negative likelihood ratios (PLR/NLR). Compared with MYC protein expression, LMO2 SP51 obtained significantly higher specificity, accuracy, PPV, and PLR (94%, 91%, 85%, and 14.33 vs 73%, 77%, 56%, and 3.26, respectively), and similar NPV and NLR (92% and 0.22 vs 95% and 0.12). Compared with LMO2 clone 1A9-1, the sensitivity of LMO2 SP51 was lower (79% vs 89%). We conclude that LMO2 SP51 may be a useful marker to screen
MYC
-R in CD10-positive LBCL.
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