Flow cytometric minimal residual disease assessment in B-cell precursor acute lymphoblastic leukaemia patients treated with CD19-targeted therapies - a EuroFlow study.

The standardized EuroFlow protocol, including CD19 as primary B-cell marker, enables highly sensitive and reliable minimal residual disease (MRD) assessment in B-cell precursor acute lymphoblastic leukaemia (BCP-ALL) patients treated with chemotherapy. We developed and validated an alternative gating strategy allowing reliable MRD analysis in BCP-ALL patients treated with CD19-targeting therapies. Concordant data were obtained in 92% of targeted therapy patients who remained CD19-positive, whereas this was 81% in patients that became (partially) CD19-negative. Nevertheless, in both groups median MRD values showed excellent correlation with the original MRD data, indicating that, despite higher interlaboratory variation, the overall MRD analysis was correct.

Comparison between immediate and delayed post space preparations: a systematic review and meta-analysis.

OBJECTIVES: This systematic review and meta-analysis compared the effects of immediate and delayed post space preparation on apical sealing. MATERIALS AND METHOD: Two independent authors conducted a systematic search (PubMed/Medline, Cochrane Library, and other databases, until February 2020) and a risk of bias evaluation. Only in vitro studies that compared the effects of immediate and delayed post space preparations on apical filling and adhesion of the post were eligible. RESULT: Of the 742 articles retrieved, 32 were included. Most of the studies used single-rooted human teeth and rotary files for root canal preparation, a single-cone technique for the filling protocol, and rotary instruments for post space preparation. Various delayed preparation times were evaluated: 8-72 h, 5-30 days (mainly 7 days), and 4 months. In nine studies, the delayed groups showed more apical leakage, while four studies reported more leakage in the immediate groups; ten studies found no significant difference. One study found more bacterial penetration in the delayed group, whereas three studies showed no significant difference. One study reported more voids in the delayed group, while another found no such difference. Three studies showed better post-bond strength in the delayed group, one in the immediate group, whereas three found no significant difference in post-bond strength. Three studies employing varied sealers for root canal fillings were considered for meta-analysis. Two subgroup analyses were also performed (one concerning the use of resin-based sealers (AH Plus), another for zinc oxide-eugenol-based sealers). The results of the meta-analysis showed that a delayed post space preparation led to a significantly higher apical leakage than an immediate preparation (mean difference = 0.41 mm, confidence interval = 0.24-0.59, p < 0.001). CONCLUSION: Delayed post space preparation seems to negatively influence apical sealing; however, further studies are needed to determine the influence of the timing of post space preparation on the other parameters. CLINICAL RELEVANCE: Immediate post space preparation may be the safest clinical choice to prevent apical leakage.

A 12-Week Randomized, Double-Blind, Placebo-Controlled Clinical Trial of Topiramate for the Treatment of Compulsive Buying Disorder.

BACKGROUND: Topiramate is an anticonvulsant that has shown promise as a pharmacological agent for the treatment of addictive disorders, including compulsive buying disorder (CBD). The aim of the present study was to examine the efficacy of topiramate in the treatment of CBD and its associated characteristics using a 12-week randomized, double-blind, placebo-controlled design. METHODS: Fifty patients seeking treatment of CBD who met the inclusion criteria were randomly assigned to either the experimental group (n = 25) or the control group (n = 25). Both groups received 4 sessions of psychoeducation. RESULTS: Forty-four participants completed the follow-up with no differences in the rate of dropout between groups. There were no differences between participants who received topiramate or placebo in reducing CBD symptoms assessed by the primary outcome scale (Yale-Brown Obsessive-Compulsive Scale - Shopping Version). However, participants who received topiramate were significantly more likely to show clinical improvement when assessed by a secondary outcome measure, the Compulsive Buying Follow-Up Scale. In addition, there was a trend among participants who received topiramate to report improvements in aspects of hoarding and impulsivity compared with the control group. There were significant improvements in comorbid depression and social adjustments over time, but no group × time interaction was found. CONCLUSIONS: The results do not provide support for the use of topiramate in the treatment of CBD. Future investigation with larger and representative samples and longer follow-up period are needed.

Standardized flow cytometry for highly sensitive MRD measurements in B-cell acute lymphoblastic leukemia.

A fully-standardized EuroFlow 8-color antibody panel and laboratory procedure was stepwise designed to measure minimal residual disease (MRD) in B-cell precursor (BCP) acute lymphoblastic leukemia (ALL) patients with a sensitivity of ≤10-5, comparable to real-time quantitative polymerase chain reaction (RQ-PCR)-based MRD detection via antigen-receptor rearrangements. Leukocyte markers and the corresponding antibodies and fluorochromes were selected based on their contribution in separating BCP-ALL cells from normal/regenerating BCP cells in multidimensional principal component analyses. After 5 multicenter design-test-evaluate-redesign phases with a total of 319 BCP-ALL patients at diagnosis, two 8-color antibody tubes were selected, which allowed separation between normal and malignant BCP cells in 99% of studied patients. These 2 tubes were tested with a new erythrocyte bulk-lysis protocol allowing acquisition of high cell numbers in 377 bone marrow follow-up samples of 178 BCP-ALL patients. Comparison with RQ-PCR-based MRD data showed a clear positive relation between the percentage concordant cases and the number of cells acquired. For those samples with >4 million cells acquired, concordant results were obtained in 93% of samples. Most discordances were clarified upon high-throughput sequencing of antigen-receptor rearrangements and blind multicenter reanalysis of flow cytometric data, resulting in an unprecedented concordance of 98% (97% for samples with MRD < 0.01%). In conclusion, the fully standardized EuroFlow BCP-ALL MRD strategy is applicable in >98% of patients with sensitivities at least similar to RQ-PCR (≤10-5), if sufficient cells (>4 × 106, preferably more) are evaluated.

Long-Term Outcomes of Childhood Acute Lymphocytic Leukemia Treated with Adapted Berlin-Frankfurt-Münster (BFM) Protocols: A Multicentric Analysis from a Developing Country.

INTRODUCTION: The objective of the current study was to determine the survival probabilities of children and adolescents with acute lymphocytic leukemia treated with adapted Berlin-Frankfurt-Münster (BFM) protocols and compare our results with the original BFM reports. METHODS: This retrospective study included 695 patients up to 19 years old treated with adapted BFM protocols between 1997 and 2018 in four hospitals in Rio de Janeiro. The 1997-2007 and 2008-2018 cohorts were analyzed separately. RESULTS: More than half of the patients were stratified into the high-risk BFM classification. Overall and event-free survivals were, in the 1997-2007 period, respectively, 88% and 80% (BFM standard risk group-SRG), 75% and 67% (intermediate risk group-IRG), and 48% and 33% (high-risk group-HRG). The corresponding numbers for the 2008-2018 period were 93% and 84% (SRG), 75% and 63% (IRG), and 64% and 57% (HRG). In the second period, both the OS (HR = 0.71, p = 0.011) and EFS (HR = 0.62, p < 0.001) were higher. Except for the intermediate-risk group, the latter results are comparable to the BFM. CONCLUSION: The BFM protocol adaptations can be safely implemented in developing countries, accounting for local specificities.

Minimal residual disease assessment in B-cell precursor acute lymphoblastic leukemia by semi-automated identification of normal hematopoietic cells: A EuroFlow study.

Presence of minimal residual disease (MRD), detected by flow cytometry, is an important prognostic biomarker in the management of B-cell precursor acute lymphoblastic leukemia (BCP-ALL). However, data-analysis remains mainly expert-dependent. In this study, we designed and validated an Automated Gating & Identification (AGI) tool for MRD analysis in BCP-ALL patients using the two tubes of the EuroFlow 8-color MRD panel. The accuracy, repeatability, and reproducibility of the AGI tool was validated in a multicenter study using bone marrow follow-up samples from 174 BCP-ALL patients, stained with the EuroFlow BCP-ALL MRD panel. In these patients, MRD was assessed both by manual analysis and by AGI tool supported analysis. Comparison of MRD levels obtained between both approaches showed a concordance rate of 83%, with comparable concordances between MRD tubes (tube 1, 2 or both), treatment received (chemotherapy versus targeted therapy) and flow cytometers (FACSCanto versus FACSLyric). After review of discordant cases by additional experts, the concordance increased to 97%. Furthermore, the AGI tool showed excellent intra-expert concordance (100%) and good inter-expert concordance (90%). In addition to MRD levels, also percentages of normal cell populations showed excellent concordance between manual and AGI tool analysis. We conclude that the AGI tool may facilitate MRD analysis using the EuroFlow BCP-ALL MRD protocol and will contribute to a more standardized and objective MRD assessment. However, appropriate training is required for the correct analysis of MRD data.

Bone Marrow Stromal Cell Regeneration Profile in Treated B-Cell Precursor Acute Lymphoblastic Leukemia Patients: Association with MRD Status and Patient Outcome.

For the last two decades, measurable residual disease (MRD) has become one of the most powerful independent prognostic factors in B-cell precursor acute lymphoblastic leukemia (BCP-ALL). However, the effect of therapy on the bone marrow (BM) microenvironment and its potential relationship with the MRD status and disease free survival (DFS) still remain to be investigated. Here we analyzed the distribution of mesenchymal stem cells (MSC) and endothelial cells (EC) in the BM of treated BCP-ALL patients, and its relationship with the BM MRD status and patient outcome. For this purpose, the BM MRD status and EC/MSC regeneration profile were analyzed by multiparameter flow cytometry (MFC) in 16 control BM (10 children; 6 adults) and 1204 BM samples from 347 children and 100 adult BCP-ALL patients studied at diagnosis (129 children; 100 adults) and follow-up (824 childhood samples; 151 adult samples). Patients were grouped into a discovery cohort (116 pediatric BCP-ALL patients; 338 samples) and two validation cohorts (74 pediatric BCP-ALL, 211 samples; and 74 adult BCP-ALL patients; 134 samples). Stromal cells (i.e., EC and MSC) were detected at relatively low frequencies in all control BM (16/16; 100%) and in most BCP-ALL follow-up samples (874/975; 90%), while they were undetected in BCP-ALL BM at diagnosis. In control BM samples, the overall percentage of EC plus MSC was higher in children than adults (p = 0.011), but with a similar EC/MSC ratio in both groups. According to the MRD status similar frequencies of both types of BM stromal cells were detected in BCP-ALL BM studied at different time points during the follow-up. Univariate analysis (including all relevant prognostic factors together with the percentage of stromal cells) performed in the discovery cohort was used to select covariates for a multivariate Cox regression model for predicting patient DFS. Of note, an increased percentage of EC (>32%) within the BCP-ALL BM stromal cell compartment at day +78 of therapy emerged as an independent unfavorable prognostic factor for DFS in childhood BCP-ALL in the discovery cohort­hazard ratio (95% confidence interval) of 2.50 (1−9.66); p = 0.05­together with the BM MRD status (p = 0.031). Further investigation of the predictive value of the combination of these two variables (%EC within stromal cells and MRD status at day +78) allowed classification of BCP-ALL into three risk groups with median DFS of: 3.9, 3.1 and 1.1 years, respectively (p = 0.001). These results were confirmed in two validation cohorts of childhood BCP-ALL (n = 74) (p = 0.001) and adult BCP-ALL (n = 40) (p = 0.004) treated at different centers. In summary, our findings suggest that an imbalanced EC/MSC ratio in BM at day +78 of therapy is associated with a shorter DFS of BCP-ALL patients, independently of their MRD status. Further prospective studies are needed to better understand the pathogenic mechanisms involved.

Successful treatment of lipoid pneumonia associated with bowel obstruction by Ascaris lumbricoides.

Partial bowel obstruction is a serious complication of ascariasis infestation generally treated with mineral oil. This prospective study aimed to evaluate the efficacy of multiple bronchoalveolar lavages (BAL) as a therapeutic strategy for reducing lung inflammation of lipoid pneumonia associated with ascariasis. The study included five children (mean age 25 months) with partial small-bowel obstruction by Ascaris lumbricoides, who underwent diagnostic bronchoalveolar lavage for assessment of refractory pneumonia. Routine biochemical, microbiological and cytological analysis were carried out in the BAL. Protein, lactate dehydrogenase and tumor necrosis factor-alpha (TNF-α) cytokine levels were determined in the serum before and after treatment. At admission, children consistently had respiratory symptoms, altered hematological function, increased immunoglobulin E serum level and peripheral blood eosinophilia. Chest tomography showed consolidation with air bronchogram (4/4), ground-glass infiltration (3/4) and decreased attenuation in the consolidation areas (2/4). Presence of marked pleocytosis with Sudan positive foamy alveolar macrophages, high protein and lactate dehydrogenase levels in the BAL indicated presence of mixed alveolitis. One child with extensive consolidation and air bronchogram in both lungs died before treatment. Multiple bronchoalveolar lavages efficiently removed alveolar oil deposits, restored BAL cellularity, improved clinical symptoms, radiological parameters and further reduced inflammatory reaction evidenced by marked decrease of the inflammatory cytokine, TNF-α. This study presents a therapeutic strategy for management of lung complications caused by mineral oil administration to treat intestinal bowel obstruction associated with ascariasis.

Flow Cytometry Immunophenotyping for Diagnostic Orientation and Classification of Pediatric Cancer Based on the EuroFlow Solid Tumor Orientation Tube (STOT).

Early diagnosis of pediatric cancer is key for adequate patient management and improved outcome. Although multiparameter flow cytometry (MFC) has proven of great utility in the diagnosis and classification of hematologic malignancies, its application to non-hematopoietic pediatric tumors remains limited. Here we designed and prospectively validated a new single eight-color antibody combination-solid tumor orientation tube, STOT-for diagnostic screening of pediatric cancer by MFC. A total of 476 samples (139 tumor mass, 138 bone marrow, 86 lymph node, 58 peripheral blood, and 55 other body fluid samples) from 296 patients with diagnostic suspicion of pediatric cancer were analyzed by MFC vs. conventional diagnostic procedures. STOT was designed after several design-test-evaluate-redesign cycles based on a large panel of monoclonal antibody combinations tested on 301 samples. In its final version, STOT consists of a single 8-color/12-marker antibody combination (CD99-CD8/numyogenin/CD4-EpCAM/CD56/GD2/smCD3-CD19/cyCD3-CD271/CD45). Prospective validation of STOT in 149 samples showed concordant results with the patient WHO/ICCC-3 diagnosis in 138/149 cases (92.6%). These included: 63/63 (100%) reactive/disease-free samples, 43/44 (98%) malignant and 4/4 (100%) benign non-hematopoietic tumors together with 28/38 (74%) leukemia/lymphoma cases; the only exception was Hodgkin lymphoma that required additional markers to be stained. In addition, STOT allowed accurate discrimination among the four most common subtypes of malignant CD45- CD56++ non-hematopoietic solid tumors: 13/13 (GD2++ numyogenin- CD271-/+ nuMyoD1- CD99- EpCAM-) neuroblastoma samples, 5/5 (GD2- numyogenin++ CD271++ nuMyoD1++ CD99-/+ EpCAM-) rhabdomyosarcomas, 2/2 (GD2-/+ numyogenin- CD271+ nuMyoD1- CD99+ EpCAM-) Ewing sarcoma family of tumors, and 7/7 (GD2- numyogenin- CD271+ nuMyoD1- CD99- EpCAM+) Wilms tumors. In summary, here we designed and validated a new standardized antibody combination and MFC assay for diagnostic screening of pediatric solid tumors that might contribute to fast and accurate diagnostic orientation and classification of pediatric cancer in routine clinical practice.

Oat bran in cardiovascular risk control in mental disorder.

OBJECTIVE: To evaluate the effect of oat bran supplementation on cardiovascular risk components of patients with mental disorders. METHOD: A before-and-after study, no control group. Cardiovascular risk indicators were assessed at baseline (M0), 90 (M1), 180 days (M2) and 180 days after supplementation (M3). RESULTS: Of the 45 patients admitted to a psychiatric institution using antipsychotics, more than two thirds had high cardiovascular risk assessed by abdominal obesity. Forty-six point seven percent were overweight and 31.1% metabolic syndrome. Oat bran was effective in reducing serum cholesterol (M0-M1), HDL-cholesterol (M1-M2), triglycerides (M1-M2), (M2-M3) and (M1-M3). In M3, there was a statistical difference for all indicators evaluated. CONCLUSION: Oat bran supplementation was effective in improving triglyceride, total cholesterol and HDL-cholesterol levels, suggesting that it is a therapeutic option for cardiovascular risk control in patients with psychiatric disorders.

Differential expression of CD73, CD86 and CD304 in normal vs. leukemic B-cell precursors and their utility as stable minimal residual disease markers in childhood B-cell precursor acute lymphoblastic leukemia.

BACKGROUND: Optimal discrimination between leukemic blasts and normal B-cell precursors (BCP) is critical for treatment monitoring in BCP acute lymphoblastic leukemia (ALL); thus identification of markers differentially expressed on normal BCP and leukemic blasts is required. METHODS: Multicenter analysis of CD73, CD86 and CD304 expression levels was performed in 282 pediatric BCP-ALL patients vs. normal bone marrow BCP, using normalized median fluorescence intensity (nMFI) values. RESULTS: CD73 was expressed at abnormally higher levels (vs. pooled normal BCP) at diagnosis in 71/108 BCP-ALL patients (66%), whereas CD304 and CD86 in 119/202 (59%) and 58/100 (58%) patients, respectively. Expression of CD304 was detected at similar percentages in common-ALL and pre-B-ALL, while found at significantly lower frequencies in pro-B-ALL. A significant association (p = 0.009) was found between CD304 expression and the presence of the ETV6-RUNX1 fusion gene. In contrast, CD304 showed an inverse association with MLL gene rearrangements (p = 0.01). The expression levels of CD73, CD86 and CD304 at day 15 after starting therapy (MRD15) were stable or higher than at diagnosis in 35/37 (95%), 40/56 (71%) and 19/41 (46%) cases investigated, respectively. This was also associated with an increased mean nMFI at MRD15 vs. diagnosis of +24 and +3 nMFI units for CD73 and CD86, respectively. In addition, gain of expression of CD73 and CD86 at MRD15 for cases that were originally negative for these markers at diagnosis was observed in 16% and 18% of cases, respectively. Of note, CD304 remained aberrantly positive in 63% of patients, despite its levels of expression decreased at follow-up in 54% of cases. CONCLUSIONS: Here we show that CD73, CD86 and CD304 are aberrantly (over)expressed in a substantial percentage of BCP-ALL patients and that their expression profile remains relatively stable early after starting therapy, supporting their potential contribution to improved MRD analysis by flow cytometry.

Altered neutrophil immunophenotypes in childhood B­cell precursor acute lymphoblastic leukemia.

An increasing number of evidences suggest a genetic predisposition in acute lymphoblastic leukemia (ALL) that might favor the occurrence of the driver genetic alterations. Such genetic background might also translate into phenotypic alterations of residual hematopoietic cells. Whether such phenotypic alterations are present in bone marrow (BM) cells from childhood B-cell precursor (BCP)-ALL remains to be investigated. Here we analyzed the immunophenotypic profile of BM and peripheral blood (PB) maturing/matured neutrophils from 118 children with BCP-ALL and their relationship with the features of the disease. Our results showed altered neutrophil phenotypes in most (77%) BCP-ALL cases. The most frequently altered marker was CD10 (53%), followed by CD33 (34%), CD13 (15%), CD15/CD65 (10%) and CD123 (7%). Of note, patients with altered neutrophil phenotypes had younger age (p = 0.03) and lower percentages of BM maturing neutrophils (p = 0.004) together with greater BM lymphocyte (p = 0.04), and mature B-cell (p = 0.03) counts. No significant association was found between an altered neutrophil phenotype and other disease features. These findings point out the potential existence of an altered residual hematopoiesis in most childhood BCP-ALL cases.

Excoriation disorder: impulsivity and its clinical associations.

Excoriation disorder is the repetitive scratching or picking of skin that leads to physical damage, distress, and functional impairment. Skin picking has been associated with impulsivity and problems with inhibition. We hypothesized that problems in these areas could be disease severity markers. We recruited 73 adults meeting DSM-5 criteria for excoriation disorder, and 50 adult controls. Those with excoriation disorder were categorized as either "high impulsive" (HI) or "low impulsive" (LI) using either a neurocognitive task of motor impulsivity (Stop Signal Task) or the Barratt Impulsiveness Scale's (BIS-11) motor impulsivity subscale. The HI subjects, based on the BIS-11, showed higher urges scores, anxiety, and depressive symptoms. These data suggest that impulsivity may reflect a specific clinical presentation among those with excoriation disorder, but the clinical characteristics differ depending upon the impulsivity measure used. Agreement on how to measure various domains of impulsivity may be important in better understanding the disorder psychopathology and so improve future treatments.

Oat bran in cardiovascular risk control in mental disorder / Salvado de avena en el control del riesgo cardiovascular en el trastorno mental / Farelo de aveia no controle de risco cardiovascular em transtorno mental

ABSTRACT Objective: To evaluate the effect of oat bran supplementation on cardiovascular risk components of patients with mental disorders. Method: A before-and-after study, no control group. Cardiovascular risk indicators were assessed at baseline (M0), 90 (M1), 180 days (M2) and 180 days after supplementation (M3). Results: Of the 45 patients admitted to a psychiatric institution using antipsychotics, more than two thirds had high cardiovascular risk assessed by abdominal obesity. Forty-six point seven percent were overweight and 31.1% metabolic syndrome. Oat bran was effective in reducing serum cholesterol (M0-M1), HDL-cholesterol (M1-M2), triglycerides (M1-M2), (M2-M3) and (M1-M3). In M3, there was a statistical difference for all indicators evaluated. Conclusion: Oat bran supplementation was effective in improving triglyceride, total cholesterol and HDL-cholesterol levels, suggesting that it is a therapeutic option for cardiovascular risk control in patients with psychiatric disorders.

RESUMEN Objetivo: Evaluar el efecto de la suplementación con salvado de avena en los componentes de riesgo cardiovascular de pacientes con trastornos mentales. Método: Un estudio antes y después, sin grupo control. Los indicadores de riesgo cardiovascular se evaluaron al inicio del estudio (M0), 90 (M1), 180 días (M2) y 180 días después de la suplementación (M3). Resultados: De los 45 pacientes ingresados en una institución psiquiátrica que usan antipsicóticos, más de dos tercios tenían un alto riesgo cardiovascular evaluado por obesidad abdominal. 46.7% tenían sobrepeso y 31.1% síndrome metabólico. El salvado de avena fue eficaz para reducir el colesterol sérico (M0-M1), el colesterol HDL (M1-M2), los triglicéridos (M1-M2), (M2-M3) y (M1-M3). En M3 hubo una diferencia estadística para todos los indicadores evaluados. Conclusión: La suplementación con salvado de avena fue efectiva para mejorarlos niveles de triglicéridos, colesterol total y colesterol HDL, lo que sugiere que es una opción terapéutica para el control del riesgo cardiovascular en pacientes con trastornos psiquiátricos.

RESUMO Objetivo: Avaliar o efeito da suplementação de farelo de aveia sobre os componentes de risco cardiovascular de pacientes com transtornos mentais. Método: Estudo antes e depois, sem grupo controle. Foram avaliados os indicadores de risco cardiovascular no início (M0), 90 (M1), 180 dias (M2) e 180 dias após a suplementação (M3). Resultados: Dos 45 pacientes internados em uma instituição psiquiátrica em uso de antipsicóticos, mais de dois terços apresentaram risco cardiovascular elevado avaliado pela obesidade abdominal. 46,7% apresentou excesso de peso e 31,1% síndrome metabólica. O farelo de aveia mostrou-se eficaz na redução do colesterol sérico (M0-M1), HDL-colesterol (M1-M2), triglicerídeos (M1-M2), (M2-M3) e (M1-M3). Em M3 houve diferença estatística para todos os indicadores avaliados. Conclusão: A suplementação do farelo de aveia foi eficaz para melhora dos níveis de triglicérides, colesterol total e HDL-colesterol, sugerindo ser uma opção terapêutica para controle dos riscos cardiovasculares de pacientes com transtornos psiquiátricos.

Accuracy of polimerase chain reaction for the diagnosis of pleural tuberculosis.

INTRODUCTION: Polymerase chain reaction (PCR)-based techniques to detect Mycobacterium tuberculosis DNA in respiratory specimens have been increasingly used to diagnose pulmonary tuberculosis. Their use in non-respiratory specimens to diagnose extrapulmonary tuberculosis is, however, controversial. In this study, we estimated the accuracy of three in-country commercialized PCR-based diagnostic techniques in pleural fluid samples for the diagnosis of pleural tuberculosis. METHODS: Patients underwent thoracenthesis for diagnosis purposes; pleural fluid aliquots were frozen and subsequently submitted to two real time PCR tests (COBAS(®)TAQMAN(®)MTB and Xpert(®)MTB/Rif) and one conventional PCR test (Detect-TB(®)). Two different reference standards were considered: probable tuberculosis (based on clinical grounds) and confirmed tuberculosis (bacteriologically or histologically). RESULTS: Ninety-three patients were included, of whom 65 with pleural tuberculosis, 35 of them confirmed. Sensitivities were 29% for COBAS(®)TAQMAN(®)MTB, 3% for Xpert(®)MTB/Rif and 3% for Detect-TB(®); specificities were 86%, 100% and 97% respectively, considering confirmed tuberculosis. Considering all cases, sensitivities were 16%, 3% and 2%, and specificities, 86%, 100%, and 97%. DISCUSSION: Compared to the 95% sensitivity of adenosine deaminase, the most sensitive test for pleural tuberculosis, the sensitivities of the three PCR-based tests were very low. We conclude that at present, there is no major place for such tests in routine clinical use.

Contribution of multiparameter flow cytometry immunophenotyping to the diagnostic screening and classification of pediatric cancer.

Pediatric cancer is a relatively rare and heterogeneous group of hematological and non-hematological malignancies which require multiple procedures for its diagnostic screening and classification. Until now, flow cytometry (FC) has not been systematically applied to the diagnostic work-up of such malignancies, particularly for solid tumors. Here we evaluated a FC panel of markers for the diagnostic screening of pediatric cancer and further classification of pediatric solid tumors. The proposed strategy aims at the differential diagnosis between tumoral vs. reactive samples, and hematological vs. non-hematological malignancies, and the subclassification of solid tumors. In total, 52 samples from 40 patients suspicious of containing tumor cells were analyzed by FC in parallel to conventional diagnostic procedures. The overall concordance rate between both approaches was of 96% (50/52 diagnostic samples), with 100% agreement for all reactive/inflammatory and non-infiltrated samples as well as for those corresponding to solid tumors (n = 35), with only two false negative cases diagnosed with Hodgkin lymphoma and anaplastic lymphoma, respectively. Moreover, clear discrimination between samples infiltrated by hematopoietic vs. non-hematopoietic tumor cells was systematically achieved. Distinct subtypes of solid tumors showed different protein expression profiles, allowing for the differential diagnosis of neuroblastoma (CD56(hi)/GD2(+)/CD81(hi)), primitive neuroectodermal tumors (CD271(hi)/CD99(+)), Wilms tumors (>1 cell population), rhabdomyosarcoma (nuMYOD1(+)/numyogenin(+)), carcinomas (CD45(-)/EpCAM(+)), germ cell tumors (CD56(+)/CD45(-)/NG2(+)/CD10(+)) and eventually also hemangiopericytomas (CD45(-)/CD34(+)). In summary, our results show that multiparameter FC provides fast and useful complementary data to routine histopathology for the diagnostic screening and classification of pediatric cancer.

Root fracture in patient in the pre-liver transplantation phase / Fratura de raiz em paciente em fase pré-transplante de fígado

ABSTRACT Liver transplantation is the treatment of choice for end-stage liver disease patients, and offers them the possibility of returning to active and longer life. A dental evaluation before transplantation is a recommended prerequisite to avoid oral infections, which can worsen the patient's overall situation. This case report refers to the root fracture of an anterior tooth in a patient who was undergoing dental treatment prior to liver transplantation. On clinical examination, the maxillary right central incisor (11) presented without color change, but with mobility in the buccolingual direction, slight flaring and extrusion, and fistula in the attached gingiva on the distal side of tooth 11. The patient reported that he had suffered a trauma in that region 30 years before. The radiographic examination showed a horizontal fracture of the middle third of the root, associated with bone damage, indicated by a radiolucent area at the distal surface of 11. The proposed treatment consisted of: scraping procedure and straightening of the distal surface, radical endodontic treatment of the coronal portion of the root, subsequent filling with Mineral Trioxide Aggregate and dental immobilization. The clinical and radiographic follow-up at three-monthly intervals allowed us to observe gradual regression of the lesion with bone formation, contributing to maintenance of the patient's general and emotional health, preserving his smile and quality of life.

RESUMO O transplante de fígado é o tratamento de escolha para o estágio final da doença hepática e oferece a possibilidade ao paciente de retorno a uma vida ativa e longa. Uma avaliação dentária antes do transplante é recomendada como pre-requisito para a eliminação de infecções orais, as quais podem piorar o quadro do paciente. Este relato de caso se refere a uma fratura radicular de dente anterior em um paciente em tratamento odontológico prévio ao transplante de fígado. No exame clínico, o incisivo central superior direito (11) apresentava-se sem alteração na coloração mas com mobilidade no sentido vestíbulo-lingual, ligeira vestibularização e extrusão, e fístula na gengiva inserida na distal do 11. O paciente relatou ter sofrido traumatismo na região há mais de 30 anos. Ao exame radiográfico, foi identificada uma linha de fratura radicular horizontal no terço médio, associada à lesão óssea indicada por uma área radiolúcida na superfície distal do 11. O tratamento proposto para o caso envolveu procedimentos de raspagem e alisamento subgengival da superfície distal, o tratamento endodôntico radical da porção coronária radicular com posterior obturação com Agregado Trióxido Mineral e imobilização dental. O acompanhamento clínico e radiográfico, em intervalos de três meses, permitiu observar a regressão gradual da lesão, com formação de osso no local, contribuindo para manutenção do quadro de saúde geral e emocional do paciente, preservando seu sorriso e sua qualidade de vida.

Análise de metaloproteases de matriz extracelular como indicador prognóstico nos gliomas malignos / Analysis of matrix metalloproteinase as prognostic indicator in malignant gliomas

Introdução: Metaloproteases de matriz extracelular(MMP) são enzimas proteolíticas sintetizadas na fase de fenótipo mais agressivo dos gliomas malignos, degradando proteínas da matriz extracelular,ocasionando ruptura da barreira hematoencefálica e contribuindo para a resposta neuroinflamatória,angiogênese e migração.Estudos mostram expressão de gelatinase A(MMP-2)proeminentemente nas células gliais tumorais,com pouca expressão na microvasculatura,enquanto expressão de gelatinase B(MMP-9)é proeminente na microvasculatrua, com porco sinal nas células tumorais.Objetivo:Neste estudo analisamos amostras de soro de 34 pacientes com gliomas malignos recidivos,antes e durante o tratamento com álcool perílico por via inalatória para determinar se a expressão de MMP poderia ser usada como indicador prognóstico.Métodos:A atividade gelatinade (MMP-2, MMP-9)nas amostras de soro foi determinada por zimografia e a atividade enzimática relativa foi determinada utilizando-se programa de análise densitométrica.Os valores foram correlacionados com exames de imagem e sobrevida dos pacientes. Resultados:Os resultados obtidos em nosso estudo evidenciaram que os pacientes com gliomas malignos apresentaram aumento da expressão de MMP-2 e MMP-9 quando comparados com pacientes saudáveis.Expressão aumentada de MMP-2 foi proporcional à progressão tumoral,sobrevida desfavorável e área de edema peritumoral.Conclusão:Esses resultados indicam proporcionalidade entre a expressão de MMP-2 e a malignidade dos gliomas, sugerindo seu emprego como indicador prognóstico para recorrência tumoral pós-operatória e sobrevida desfavorável dos pacientes.