Search

Irma Olarte Carrillo,1 Christian Omar Ramos Peñafiel,2 Iveth Mendoza Salas,2 Rafael Cerón Maldonado,1 Anel Irais García Laguna,1 Adrián De la Cruz Rosas,1 Carlos Martinez Murrillo,2 Adolfo Martinez Tovar1 1Laboratorio de Biología Molecular, Servicio de Hematología, Hospital General de México, “Dr. Eduardo Liceaga”, Ciudad de México, Mexico; 2Servicio de Hematología, Hospital General de México, “Dr. Eduardo Liceaga”, Ciudad de México, MexicoCorrespondence: Adolfo Martinez Tovar, Labortororio de Biología Molecular, Servicio de Hematología, Hospital General de Mexico “Dr. Eduardo Liceaga”, Ciudad de México, Mexico, Email mtadolfo73@hotmail.comObjective: In the last two years progress was made in molecular, physio pathological understanding and the form of transmission of COVID-19, and different therapeutic strategies have been explored to deal with the situation of the pandemic. However, the evaluation of certain genes that participate in the metabolism and transport of these drugs has not been fully explored. A lack of response to treatment and a lower survival have been observed that may be due to the presence of the ABCB1 drug resistance gene. Our research group analyzed whether the expression levels of the ABCB1 gene are associated with comorbidities, treatments, overall survival and risk of death in patients with severe COVID-19.Methods: The expression levels of the ABCB1 gene were analyzed by RT-qPCR in 61 patients diagnosed with COVID-19. The association between the levels of expression, the risk variables and different treatments were determined by the Chi-Square test and the Fisher’s exact test. Global Survival (GS) was determined by the Kaplan–Meier method. The impact of high levels of expression and the risk of death was performed by odds ratio.Results: The different risk variables showed that patients with either high or absent levels of ABCB1 gene expression presented a greater risk of death (OR 3.08, 95%, CI 1.02– 9.26) as well as need for ventilatory support (OR 2.8, 95%, CI 0.98 − 8.5). Patients with diabetes and COVID-19, treated with metformin, were associated with a lower risk of death (OR 1.11, 95%, CI 0.38– 3.22). OS with respect to high or absent levels of expression of the ABCB1 gene was lower.Conclusion: High levels or null expression of the ABCB1 gene are associated with a higher risk of death or progression of the disease, the use of metformin in patients with COVID-19 confers a lower risk of death.Keywords: ABCB1, SARS-CoV2, drug transporter genes

Purpose: Despite advances in the treatment of adult acute lymphoblastic leukemia (ALL), relapse remains the most significant challenge in improving prognosis. Measurable residual disease (MRD) assessment can predict bone marrow relapse based on MRD positivity. As access to innovative therapies remains limited because of the high cost, chemotherapy is the widely utilized treatment option. The efficacy of a combination of bortezomib and Hyper-CVAD has been reported in patients with multiple myeloma; however, its efficacy has not yet been confirmed in patients with ALL., Methods: This prospective cohort study involved patients with ALL who presented with MRD-positive results or relapse and received treatment with a combination of bortezomib and Hyper-CVAD at two reference centers in Mexico City., Results: Of the 20 patients with positive MRD included in this study, 60% (n = 12) exhibited MRD negative results after combination treatment, 30% (n = 6) persisted positive MRD results, and 10% (n = 2) passed away. Of the 23 patients with bone marrow relapse, 43.5% (n = 10) achieved a second complete remission (2CR), 34.8% (n = 6) exhibited refractory status, and 21.7% (n = 5) passed away. To achieve a 2CR, 20% (n = 2) patients required less than four cycles of treatment, 50% (n = 5) required four cycles (two A and B cycles each), and 30% (n = 3) required six cycles., Conclusion: The combination of bortezomib and Hyper-CVAD treatment exhibited better results in achieving MRD negative results, indicating its potential as a promising first-line treatment strategy for ALL., Competing Interests: Declarations. Consent for publication: Written informed consent was obtained from all the participants. The Biosecurity, Ethics and Research Committee of “Hospital General de México “Dr. Eduardo Liceaga” approved the informed consent. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Acute lymphoblastic leukemia (ALL) represents around 25% of adult acute leukemias. Despite the increasing improvement in the survival rate of ALL patients during the last decade, the heterogeneous clinical and molecular features of this malignancy still represent a major challenge for treatment and achieving better outcomes. To identify aberrantly expressed genes in bone marrow (BM) samples from adults with ALL, transcriptomic analysis was performed using Affymetrix Human Transcriptome Array 2.0 (HTA 2.0). Differentially expressed genes (DEGs) (±2-fold change, p -value < 0.05, and FDR < 0.05) were detected using the Transcriptome Analysis Console. Gene Ontology (GO), Database for Annotation, Visualization, and Integrated Discovery (DAVID), and Ingenuity Pathway Analysis (IPA) were employed to identify gene function and define the enriched pathways of DEGs. The protein-protein interactions (PPIs) of DEGs were constructed. A total of 871 genes were differentially expressed, and DNTT , MYB , EBF1 , SOX4 , and ERG were the top five up-regulated genes. Meanwhile, the top five down-regulated genes were PTGS2 , PPBP , ADGRE3 , LUCAT1 , and VCAN . An association between ERG , CDK6 , and SOX4 expression levels and the probability of relapse and death was observed. Regulation of the immune system, immune response, cellular response to stimulus, as well as apoptosis signaling, inflammation mediated by chemokines and cytokines, and T cell activation were among the most altered biological processes and pathways, respectively. Transcriptome analysis of ALL in adults reveals a group of genes consistently associated with hematological malignancies and underscores their relevance in the development of ALL in adults.

IL-15 is a proinflammatory myokine essential for activating NK cells and CD8+ T lymphocytes, and its overexpression has been related to reducing overall survivorship in patients with acute lymphoblastic leukemia (ALL). Physical exercise has been shown to be safe, feasible, and beneficial in hematological cancers. Exercise requires the activation of muscles that secrete cytokines, such as IL-15, causing immune mobilization. The objective was to compare the outcomes of two training routines on IL-15 and survival prognosis in adult patients diagnosed with ALL. A blind randomized clinical study was carried out where twenty-three peripheral blood samples were obtained pre and postexercise intervention from patients categorized into three types of intervention: the resistance exercise group (REG), the cross-training exercise group (CEG), and the control group (CG). Changes in IL-15 levels during the intervention were not significant in any of the groups (CG p = 0.237, REG p = 0.866, and CEG p = 0.678). However, 87.5% of patients who received an exercise intervention achieved remission, while only 21.73% experienced a relapse. There were no deaths during the study. Although IL-15 level adaptation in the REG and the CG performed similarly, the REG induced a better clinical outcome. Resistance exercises may help improve survival prognosis and reduce relapses in patients with ALL.

Patients with acute lymphoblastic leukemia (ALL) undergoing induction decrease their physical capacity, lose muscle mass, and decrease their quality of life (QOL). The safety, feasibility, and benefits of exercise during chemotherapy have been proven, but the effects of cross-training activities have yet to be analyzed. To measure the effects of cross-training on body composition, physical performance, and QOL, a blind randomized clinical trial was carried out. A total of 33 patients were included and randomized into a cross-training exercise group (CEG), a resistance exercise group (REG), and a control group (CG). During induction, patients received an exercise routine three to five days a week for 30 to 50 min each. Body composition, QOL, and physical performance were measured at baseline, up to discharge, and at a follow-up of two months. Body composition improved in the REG and CEG. In the CG, muscle mass decreased and fat mass increased ( p = 0.020 and 0.020, respectively). The REG and CEG had significant positive improvements in physical performance compared to the CG. QOL showed no differences in any group ( p = 0.340). Cross-training and resistance exercise are essential to improve body composition and physical performance during induction. Considering the prognostic value of physical performance, we propose integrated training exercises as adjuvant therapy in adult patients with ALL.

Background: Obesity has been associated with a low-grade proinflammatory state, and it has been related to the development of cancer in general, including hematologic cancer., Aim: The present work aimed to identify the association of the diagnosis of obesity according to the body mass index (BMI) with prognostic factors of adult patients with Acute Lymphoblastic Leukemia (ALL)., Patients and Method: This observational, retrospective study included hospitalized patients diagnosed with ALL of the B-cell lineages. BMI was estimated based on the weight and height registered on clinical records at the admission of the patients. The relapse risk and bone marrow relapse were determined, and the survival rate was measured. The statistical analysis included the Kaplan-Meier method using the log-Rank test., Results: This study included 128 clinical records of patients. Weight had no significant association with relapse risk. The frequency of bone marrow relapse was 43.8%. Obesity did not impact overall survival (p = 0.640) or disease-free survival (p = 0.527). The presence of obesity does not behave as a relapse risk variable (p = 0.873). BMI with a 30 kg/m2 cut-off point did not influence relapse risk (OR 1.078)., Conclusion: Obesity is not an independent risk factor for the prognosis of adult patients with Acute Lymphoblastic Leukemia B-lineage.

Background: Leukemia is a neoplasm with high incidence and mortality rates. Mitotic death has been observed in tumor cells treated with chemotherapeutic agents. Ras family proteins participate in the transduction of signals involved in different processes, such as proliferation, differentiation, survival, and paradoxically, initiation of cell death., Methods: This study investigated the effect of H-Ras expression on human T-cell acute lymphoblastic leukemia MOLT-4 cells. Cells were electroporated with either wild-type (Ras wt ) or oncogenic mutant in codon 12 exon 1 (Ras mut ) versions of H-Ras gene and stained for morphological analysis. Cell viability was assessed using trypan blue staining and cell cycle analysis using flow cytometry. H-Ras gene expression was determined using quantitative real-time reverse transcription polymerase chain reaction. The t , ANOVA, and Scheffe tests were used for statistical analysis., Results: Human T-cell acute lymphoblastic leukemia MOLT-4 cells showed nuclear fragmentation and presence of multiple nuclei and micronuclei after transfection with either wt or mutant H-Ras genes. Cell cycle analysis revealed a statistically significant increase in cells in the S phase when transfected with either wt (83.67%, P <0.0005) or mutated (81.79%, P <0.0001) H-Ras genes. Although similar effects for both versions of H-Ras were found, cells transfected with the mutated version died at 120 h of mitotic catastrophe., Conclusion: Transfection of human T-cell acute lymphoblastic leukemia MOLT-4 cells with either normal or mutated H-Ras genes induced alterations in morphology, arrest in the S phase, and death by mitotic catastrophe.

No disponible.

Background: Acute myeloid leukemia (AML) is a heterogeneous hematologic malignancy characterized by different genetic alterations that cause changes in the normal mechanisms of differentiation, which are associated with chemoresistance. The ABCB1 gene is part of a family of ATP-binding cassette (ABC) transporter genes involved in the progression of various types of cancer. The following work aimed to evaluate the expression levels of the ABCB1 gene and the C3435T SNP with the response to first-line treatment and survival in patients with AML., Methods: In total 135 samples were taken to isolate total RNA and DNA at the beginning of the treatment. Expression analysis by RT-qPCR and SNP C3435T assessment method were performed for real-time Polymerase chain reaction (qPCR)., Results: The expression levels impact on the survival of patients with AML compared to low or absent levels; the CC genotype was found in 22.9%, the CT genotype was found in 47.4%, and the TT genotype was found in 29.6%, the presence of the C3435T SNP, the TT genotype also impacts with a lower survival compared to CT and CC genotypes. In addition, it was shown that the dominant model significantly impacts survival., Conclusion: In conclusion, we have found that the overexpression of the ABCB1 gene, as well as the presence of the TT genotype of the C3435T SNP, contributes to a worse prognosis in AML., (© 2021. The Author(s).)

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is the cause of the coronavirus disease 2019 (COVID-19) pandemic, which has a high case fatality rate. Most severely ill patients develop a special type of coagulopathy that had not been described before and that is now considered the main cause of death. For this reason, anticoagulant treatment has become one of the cornerstones of the treatment of this infection. However, the rate at which the evidence regarding the use of anticoagulants is generated is quite fast, and sometimes it is difficult to interpret and conflicting. After having performed an extensive review of the published literature, this proposal for the use of anticoagulant treatment is made, taking into account available resources in Mexico., (Copyright: © 2021 Permanyer.)

Introduction: Various biomarkers based on blood counts have been useful for the prognosis of patients critically ill with COVID-19., Objective: To describe the usefulness of the neutrophil-to-lymphocyte (NLR), monocyte-to-lymphocyte (MLR) and lymphocyte-to-platelet (LPR) ratios for the prognosis of mortality and ventilatory support requirement for COVID-19., Method: Retrospective cohort of clinical records of patients with COVID-19 who required hospital care., Results: One-hundred and -twenty-five cases were analyzed; mean age was 51 years, and 60 % were of the male gender; 21.6 % had type 2 diabetes mellitus, and 18.4 % had hypertension. Mean leukocyte count was 9.5 x 10 3 /µL, with a neutrophil mean of 8.0 x 10 3 /µL. Mean NLR was 12.01, while for MLR it was 0.442, and for LPR, 373.07. Regarding the area under the curve, the following values were recorded for mortality: 0.594 for NLR, 0.628 for MLR and 0.505 for LPR; as for mechanical ventilation, the values were 0.581 for NLR, 0.619 for MLR and 0.547 for LPR. In the univariate analysis, an NLR value > 13 (OR: 2.750, p = 0.001) and an MLR of > 0.5 (OR: 2.069, p = 0.047) were associated with mortality; LPR showed no impact on mortality or respiratory support., Conclusion: NLR and MLR are useful for predicting mortality in patients with COVID-19., (Copyright: © 2019 Permanyer.)

Acute lymphoblastic leukemia (ALL) is a hematologic malignancy characterized by the clonal expansion of hematopoietic lymphoid progenitors. With new target therapies, the survival of adults with ALL has improved in the past few decades. Unfortunately, there are no large ALL patient series in many Latin American countries. Data from the Acute Leukemia Workgroup that includes five Mexico City referral centers were used. Survival was estimated for adult patients with ALL during 2009-2015. In total, 559 adults with ALL were included. The median age was 28 years; 67% were classified into the adolescent and young adult group. Cytogenetic information was available in 54.5% of cases. Of the 305 analyzed cases, most had a normal caryotype (70.5%) and Philadelphia-positive was present in 16.7%. The most commonly used treatment regimen was hyper-CVAD. In approximately 20% of cases, there was considerable delay in the administration of chemotherapy. Primarily refractory cases accounted for 13.1% of patients. At the time of analysis, 26.7% of cases had survived. The 3-year overall survival was 22.1%. The main cause of death was disease progression in 228 (55.6%). Clinical and public health strategies are needed to improve diagnosis, treatment and survivorship care for adult with ALL. This multicentric report represents the largest series in Mexico of adult ALL patients in which a survival analysis and risk identification were obtained., (© 2018 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.)

Introduction: In developing countries, there is commonly a lack of population-based cancer registries or underreporting, thus not recognizing the true dimensions of the problem., Aim: To describe the age and sex frequencies of the major subtypes of leukemias in two hospitals of reference in the metropolitan area of Mexico City., Material and Methods: This is a descriptive and retrospective study, based on medical records of two hematology services during January 2007 to October 2014; all cases diagnosed with leukemia were included., Results: A total of 1,432 cases were included with a median age of 38 years (range, two months to 115 years). There were significant age differences between subtypes of leukemia (ANOVA test, p = 0.000): chronic lymphocytic with a mean age of 64.8 years, higher than chronic myeloid (43.4 years) and all acute leukemias (lymphoblastic: 32.6 years, myeloblastic 43.5 years). Of the patients, 51.8% (n = 742) were women, although males predominated in chronic myeloid (57.8%) and lymphocytic (60%) leukemia. Acute lymphoblastic leukemia was the more common variety, FABL2 subtype, followed by myeloid leukemia M4, M2, and chronic myeloid., Conclusions: It is necessary to develop inter-institutional works in order to group data of different population sectors and improve the epidemiological profile of leukemia in Mexico.

Background: The incidence of acute leukemia (AL) has increased. Its prognosis is variable and depends on several baseline characteristics with a highly heterogeneous presentation. In Mexico, large-scale descriptive studies have not yet been published; the objective of this study was to analyze the initial basic characteristics of patients diagnosed with AL in our population., Patients and Methods: In this multicenter, retrospective study, 1018 patients ≥ 16 years of age and diagnosed with AL between 2009 and 2014, were included. We described age, gender, complete blood count, and AL subtype according to flow cytometry analysis., Results: Acute lymphoblastic leukemia (ALL) was as common as acute myeloid leukemia (AML) (51% vs. 49%). The median age was 31 years. Only 9.6% of patients with ALL were positive for the Philadelphia chromosome. No gender differences were observed. The median age at presentation of AML was 43 years. Acute promyelocytic leukemia was the most frequent AML subtype (38.3%), with a median age of 37 years., Conclusion: ALL is equally as frequent as AML in patients ≥16 years of age. Philadelphia-positive prevalence is less frequent than that reported in literature. AML cases occur in a younger age in comparison with other countries. There is a higher rate of acute promyelocytic leukemia among our patients compared with other non-Latin American populations. This study is the largest ever performed in Mexico regarding descriptive AL data., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Introduction: A lack of a population-based cancer registry or underreporting is common in developing countries, without knowledge of the true dimensions of the problem., Aim: To describe the age and sex frequencies of the major subtypes of leukemias in two reference hospitals in the metropolitan area of Mexico City., Material and Methods: A descriptive and retrospective study, based on medical records of two hematology services during January 2007 to October 2014; all cases diagnosed with leukemia were included., Results: A total of 1,432 cases were included, with a median age of 38 years old (2 months to 115 years). There were significant age differences between subtypes of leukemia (ANOVA test, p = 0.000); chronic lymphocytic with a mean age of 64.8 years, higher than chronic myeloid (43.4 years) and all acute leukemias (lymphoblastic: 32.6 years, myeloblastic 43.5 years). Of the patients, 51.8% (n = 742) were women, although males predominated in chronic myeloid (57.8%) and lymphocytic (60%) leukemia. Acute lymphoblastic leukemia was the more common variety, L2 subtype of the French-American-British classification, followed by myeloid leukemia M4, M2, and chronic myeloid., Conclusions: it is necessary to develop inter-institutional works in order to group data of different population sectors and improve the epidemiological profile of leukemias in Mexico.

Objetive: To know the surgical-pathologic correlation to assess the state of the edges in wide local excisions of breast cancer in clinical stages., Material and Methods: retrospective and descriptive study, conducted in Breast Tumors Unit from Oncology Service of the General Hospital of Mexico, in the period from January 2009 to December 2011, with follow-up in December2012. Were included patients with breast cancer in early clinical stages, subject wide local excisions histopalogic report of a second surgery., Results: From wide local excisions, 119 (28.5%) were due to breast cancer and included. Positive margins after initial surgery were diagnosed in 63 patients (52.9%). The residual tumor found in the second surgery was 39.7%. The variables associated with the presence of positive margins and statistically significant (p≤ 0.05) were: multicentricity, tumor size clinical and pathological, histological subtypes, and tumor grade. The age and clinical stage were not statistically significant. The variables associated with the presence of residual tumor and are statistical relevance (p≤ 0.05) were clinical stage, tumor size, clinical and pathological, histological variant and histological grade. Age and multicentricity were not associated with the presence of residual tumor., Conclusion: Although each case must be individualized, these results demonstrate the analyzed factors must be taken into account during the planning of breast conservative procedures, and despite an histopalogical report of margin after an initial surgery, even seconds procedures can be performed to conserve the organ.

In order to assess the mortality and toxicity of the Hyper-CVAD protocol used as first-line treatment of acute lymphoblastic leukemia, a retrospective cohort study was performed in patients less than 40 years of age from March to September 2011 treated with Hyper-CVAD regimen. Mortality and toxicity was compared with the results of patients treated with the institutional HGMLAL07 regimen between 2009-2012. 18 patients were included; the median age was 26 years old. Complete remissions (67.7% versus 81.9%) as well as one-year (40% versus 62%) and 2 year survival rates (18% versus 34%) were lower with the Hyper-CVAD regimen. By selecting only patients younger than 35 years, the effectiveness of Hyper-CVAD was also lower. In our experience and because of its high cost and toxicity, the Hyper-CVAD regimen should be limited to patients with relapsed or refractory leukemia.

Background: Recently it has been reported a benefit effect with the use of metformin in patients with malignant disease. Our objective was to evaluate the effect of adding metformin to chemotherapy regimen over the percentage of early relapse in acute lymphoblastic leukemia., Methods: A prospective, longitudinal and experimental study was performed in patients with de novo acute lymphoblastic leukemia enrolled in the Hospital General de México. They were divided in two groups: first group received chemotherapy + metformin (850 mg three times a day); second group only received standard chemotherapy. The sample was randomized 3:1 in favor of the second group., Results: 93 patients were included (73 treated with chemotherapy + metformin and 20 received standard chemotherapy), with 303 ± 53 days of follow-up. Complete remission was higher in the group without metformin (81.3 % [n = 61] versus 70 % [n = 14]), which also presented more patients with relapse (47.9 % versus 25 %). Overall survival at one year was of 68 % and free survival disease was 64 %, without significant differences between groups. Absence of metformin was the only variable of adverse prognostic considered significant (p = 0.55). Cox regression showed that adding metfomin reduced 56 % the risk of relapse., Conclusions: The adding metformin to the treatment of leukemias showed that was useful in our research. However, randomized and double-blind studies must be designed in order to express final recommendations about its use.

Background: The nasal colonization by Staphylococcal (epidermidis or aureus) is frequent and it has importance when it is associated to bacteremia in immunocompromised patients. The objective was to determine the frequency of strains that colonize the nasal mucosa in patients with leukemia and its relationship with peripheral blood cultures., Methods: A retrospective, observational, transversal, retrolective study was done. We analyzed the weekly results of nasal cultures and peripheral blood cultures in patients with leukemia undergoing chemotherapy. The chi-squared test and odds ratio value were estimated in the statistical analysis., Results: We included 67 patients, 55 of them with acute lymphocytic leukemia (ALL); 28.5 % of the cultures (n = 47) corresponded to a positive nasal culture. Staphylococcus epidermidis and Staphylococcus aureus were the most isolated bacteria. During the first week of treatment, the positive cultures were the most frequently. All the samples isolated were sensitive to vancomycin or linezolid. It was established only the association between negative nasal cultures and negative peripheral blood cultures (p = 0.0005). Odds ratio for positive nasal cultures and the risk of bacteremia was 0.0269., Conclusions: The frequency of the positive bacteria culture was low, with an adequate sensitivity measure. The presence of bacteria in nasal culture was not identified as a risk factor for the occurrence of bacteremia.

Background: lymphoma is the most frequent lymphoid neoplasm in our country. Its diagnosis is based on histopathological findings. The lymph node imprint has been used for more than 40 years. The aim was to establish the sensitivity, specificity, positive predictive value and negative predictive value of lymph node imprint and estimate the inter-observer rate., Methods: we did an observational, retrospective, prolective study, based on the lymph node imprint obtained by excisional biopsies over a period of 6 years., Results: the inclusion criteria was met on 199 samples, 27.1 % were considered as reactive (n = 54), 16.1 % Hodgkin lymphoma (n = 32), 40.2 % (n = 80) non-Hodgkin lymphoma and 16.6 % (n = 33) as metastatic carcinoma. Comparing with the final histopathology report, the sensitivity and specificity of lymph node imprint were 88 % (0.81-0.95) and 64 % (0.55-0.73) respectively, the positive predictive value was 67 % (0.59-0.76) and the negative predictive value was 86 % (0.79-0.94). The interobserver kappa index was 0.467., Conclusions: the lymph node imprint remains as a useful tool for the diagnosis of lymphoid neoplasm. The agreement between observers was acceptable.

Objective: to compare the frequency of acute leukemia in two periods of study in a reference Hospital at Mexico City., Methods: it was an observational study. There were registered 250 cases with the morphology diagnostic criterion of acute leukemia., Results: 63 cases versus 92 of acute lymphoid leukemia (ALL) and 16 versus 79 acute myeloid leukemia (AML) of 1990-1992 versus 2008-2009 respectively was observed; corresponding to 62 % in total of ALL in both periods and 38 % of AML. The ALL was the most frequent variant (62 % versus 38 %). The median age was 26 years, male 52 %. It showed a significant increase in the number of admissions in the period of 2008-2009 and acute promyelocytic leukemia also showed an increase (p = 0.001)., Conclusions: acute leukemia is the main cause of admission, mortality and morbidity. Our study differs slightly from the literature; the leukemia that diagnostic with major frequency was the acute lymphoid leukemia increasing his number in the period of 2008-2009. The significant rise due to an increase use of new diagnostic tools such as the flow cytometry and molecular biology.