Discriminant analysis and machine learning approach for evaluating and improving the performance of immunohistochemical algorithms for COO classification of DLBCL.

BACKGROUND: Diffuse large B-cell lymphoma (DLBCL) is classified into germinal center-like (GCB) and non-germinal center-like (non-GCB) cell-of-origin groups, entities driven by different oncogenic pathways with different clinical outcomes. DLBCL classification by immunohistochemistry (IHC)-based decision tree algorithms is a simpler reported technique than gene expression profiling (GEP). There is a significant discrepancy between IHC-decision tree algorithms when they are compared to GEP. METHODS: To address these inconsistencies, we applied the machine learning approach considering the same combinations of antibodies as in IHC-decision tree algorithms. Immunohistochemistry data from a public DLBCL database was used to perform comparisons among IHC-decision tree algorithms, and the machine learning structures based on Bayesian, Bayesian simple, Naïve Bayesian, artificial neural networks, and support vector machine to show the best diagnostic model. We implemented the linear discriminant analysis over the complete database, detecting a higher influence of BCL6 antibody for GCB classification and MUM1 for non-GCB classification. RESULTS: The classifier with the highest metrics was the four antibody-based Perfecto-Villela (PV) algorithm with 0.94 accuracy, 0.93 specificity, and 0.95 sensitivity, with a perfect agreement with GEP (κ = 0.88, P < 0.001). After training, a sample of 49 Mexican-mestizo DLBCL patient data was classified by COO for the first time in a testing trial. CONCLUSIONS: Harnessing all the available immunohistochemical data without reliance on the order of examination or cut-off value, we conclude that our PV machine learning algorithm outperforms Hans and other IHC-decision tree algorithms currently in use and represents an affordable and time-saving alternative for DLBCL cell-of-origin identification.

Plasma phospholipid fatty acids in obese male and female Mexican children.

AIM: To assess the plasma phospholipid (PL) fatty acid composition in obese Mexican children and evaluate gender differences and predisposition to health risks. METHODS: This was a case-control study of 100 obese and 100 normal-weight children aged 6-12 years. BMI, waist circumference, triceps skinfold, and percentage of body fat were determined. PL fatty acids were measured by gas chromatography. RESULTS: Compared to normal-weight children, obese children had significantly higher proportions of 18:0, 16:1n-7, 20:3n-6, saturated fatty acids (SFA), total highly unsaturated fatty acids (UFAs), and total n-3 but lower proportions of 18:1n-9, 18:2n-6, 22:5n-6, monounsaturated fatty acids, and total n-9. Obese children had significantly higher ratios of 20:3n-6/18:2n-6 and 18:3n-6 + 20:3n-6/18:2n-6, and they had lower ratios of 20:4n-6/20:3n-6 and n-6/n-3. The predictors positively associated with obesity, in decreasing association strength, were 16:1n-7, 20:3n-6, and SFA. Obese boys showed significantly higher 20:4n-6, 20:5n-3, 20:3n-6/18:2 n-6, 18:3n-6 + 20:3n-6/18:2n-6, and 20:5n-3/20:4n-6, higher UFAs, and lower 18:2n-6 and 22:6n-3/20:4n-6 compared to obese girls (p < 0.05). CONCLUSION: Significant alterations in the proportions of plasma PL fatty acids were found in obese children, especially in male subjects, which might place them in danger of early cardiovascular risk; however, an insulin-resistant state might be responsible for their fatty acid composition. More studies are needed since there are none in Mexican children.

Induction of apoptosis and effect on CD20+ using rituximab on autologous peripheral blood stem cell harvests from patients with B cell lymphomas.

Purging of neoplastic cells for autologous stem cell transplantation is usually done in vivo by administering chemotherapy and/or other agents before harvesting. It is also possible to decrease malignant cells counts directly in the cell harvest. In this study, we ascertained the effect of anti-CD20 monoclonal antibody and rituximab administration on peripheral blood hematopoietic stem cells. Five samples of stem cell harvests from different patients with B cell lymphoma were obtained. Each sample was divided in two tubes with calcium gluconate (20 mEq/50 microl). Rituximab (1 mg/600,000 mononuclear cells) was added to one of the tubes. Using flow cytometry, CD19, CD20 (B cell markers), and CD95 (apoptosis marker), expression was measured at baseline and 24 h after the addition of rituximab. A one-sided t-test with equal variances was used to analyze the results. Immediately after rituximab addition, CD20 expression became null. No significant difference in variation of CD19 expression was detected after the addition of rituximab (-3.64% control vs. 0.63% rituximab, p = 0.69). Mean variations of percentage of CD95 expression were 2.9% (controls) and 10.52% (rituximab tubes) (p = 0.06). We conclude that rituximab is capable of initiating apoptosis in vitro. We found no decrease in the CD19+ cell count, used as a surrogate marker for CD20+ cells, meaning that, at least in 24 h, apoptosis activation is not capable of decreasing CD20+ cell numbers. In vitro purging of peripheral blood stem cells harvests with rituximab could be part of a broader therapeutic strategy to be offered to lymphoproliferative disorder patients.

A randomized trial of dexamethasone before remission induction, in de novo childhood acute lymphoblastic leukemia.

We evaluated the impact of adding dexamethasone before chemotherapy in 95 children with de novo standard-risk acute lymphoblastic leukemia (ALL). The children were randomly divided into 2 groups: one group was given dexamethasone, the other was not. The initial characteristics and mean follow-up of both groups were similar. Day +14 blast percentage was significantly lower in the dexamethasone group. Disease-free survival at 40-months follow-up was better (almost significantly so) in the dexamethasone group.

[Autologous bone marrow transplantation as a treatment for autoimmune disease: mechanisms and results]. / Trasplante autólogo de médula ósea como tratamiento de enfermedades autoinmunes: mecanismos y resultados.

Autoimmune diseases are characterized by immune response against self antigens. One of the current research interests in this field is oriented toward development of tolerance. One of the newest options in the search for tolerance is autologous bone marrow transplantation: a variant of bone marrow transplant in which the patient's own hematopoietic stem cells are reinfused after myeloablative therapy. The idea of using bone marrow transplant in treatment of autoimmune diseases derived from observing remission in autoimmune diseases in patients transplanted due to coexisting neoplastic disease. Although an isolated initial report of bone marrow transplant as treatment for autoimmune disease questioned the utility of this procedure, over all, results are encouraging. To compile information in a programmed and systematic manner, it is necessary to send more patients in all stages of immune diseases to specialized centers to be included in large multicenter randomized trials. In time, the role for this procedure in autoimmune diseases will become clear.

Molecular detection and prognostic value of epithelial markers mRNA expression in peripheral blood of advanced non-small cell lung cancer patients.

BACKGROUND: Few studies, have evaluated the prognostic impact of the quantification of mRNA expression levels in advanced non-small cell lung cancer (NSCLC). OBJECTIVE: The aim of this work was to quantify mRNA expression levels in peripheral blood through three epithelial markers in patients with stages IIIB and IV in NSCLC. METHODS: Seventy advanced NSCLC patients and ten healthy controls were included. All patients received platinum-based chemotherapy in first line treatment. Peripheral blood was obtained of each participant and mRNA expression levels present in circulating cells were quantified by molecular techniques (RT-PCR) using three epithelial markers: cytokeratin (CK)-18, CK-19 and Carcinoembryonic-Antigen (CEA). The expression levels were quantified from a standard curve using the cDNA obtained from A549 cells. Registered in ClinicalTrials.gov (NCT01052818). RESULTS: We found a significant statistical correlation between levels of CK-18, CK-19 and CEA mRNA. mRNA expression levels were lower in patients who present three or less metastasis; higher CEA mRNA expression was associated a worse progression-free survival to platinum-based chemotherapy and overall survival. CONCLUSION: RNA expression of CEA by RT-PCR is useful as a prognostic marker in advanced NSCLC.

Clinical and pathological characteristics, outcome and mutational profiles regarding non-small-cell lung cancer related to wood-smoke exposure.

HYPOTHESIS: Although smoking is the major risk factor for non-small-cell lung cancer (NSCLC), other factors are also associated with lung carcinogenesis, such as wood-smoke exposure (WSE). This article has been aimed at suggesting that lung cancer related to cigarette smoking and lung cancer related to WSE have different clinical and genetic characteristics. EXPERIMENTAL DESIGN: A cohort of 914 lung cancer patients was prospectively studied; they had been treated at Mexico's National Cancer Institute between 2007 and 2010. The associations of WSE and cigarette smoking with clinical characteristics, mutation profile, response to chemotherapy, and epidermal growth factor receptor tyrosine kinase inhibitors were analyzed, and overall survival (OS) rate was calculated. The trial was registered with ClinicalTrials.gov: NCT01023828. RESULTS: Of the lung cancer patients studied, 95.1% were classified as coming within the NSCLC histology subtype; 58% of the patients smoked cigarettes, 35% had a background of WSE (exposure to both cigarette smoke and wood smoke was documented in 12.1% of all patients), and 19.4% patients had no smoke-exposure background. WSE was associated with NSCLC and adenocarcinoma histology, and was also more frequently associated with epidermal growth factor receptor-mutations than cigarette-smoking patients were (50.0% cf. 19.4%), whereas KRAS mutations were less common in WSE patients (6.7%) than in smokers (21%). WSE patients had a higher epidermal growth factor receptor tyrosine kinase inhibitor response rate (39.7%) than smokers (18.8%). The NSCLC patient WSE group's OS was longer (22.7 months) than that for smokers (13.8 months). CONCLUSION: NSCLC patients who smoked tobacco/cigarettes differed from those having a background of WSE regarding tumor histology, mutation profile, response rate, and OS, indicating that different carcinogenic mechanisms were induced by these two types of smoke exposure.

Metabolic syndrome risk factors among a sample of overweight and obese Mexican children.

The objective of this study was to estimate the prevalence and correlations of components of the metabolic syndrome (MetS) using the International Diabetes Federation (IDF) pediatric definition in a cross-sectional study of 215 overweight/obese Mexican children aged 6 to 12. There are no previous studies of this kind in Mexican children. Clinical, anthropometric, and laboratory measurements were performed. The prevalence of MetS using the pediatric IDF criteria was 6.7% (95% confidence interval, 4.0-11.1). A higher proportion of children in the younger age group had waist circumference above the cutoff, while a higher proportion in the older age group had hyperglycemia. Children with MetS had higher percentages of body fat, body mass index, total cholesterol, and low-density lipoprotein cholesterol. Increased triglycerides, decreased high-density lipoprotein cholesterol, and waist circumference were most highly associated with MetS. This has significant implications for public health.

Serum 25-hydroxyvitamin d concentration, life factors and obesity in Mexican children.

Although current evidence emphasizes a high prevalence of vitamin D deficiency and an inverse association between serum 25-hydroxyvitamin D (25-OHD) concentration and obesity, no studies have been conducted in Mexican children. The objective was to determine the prevalence of vitamin D deficiency and its association with obesity and lifestyle factors in a sample of school-aged Mexican children. A cross-sectional study of 99 obese and 99 nonobese 6-12 year-old children, skin phototypes III-V, from six public schools was conducted during summer at latitude 25 degrees 40', in northeastern Mexico. Anthropometric measurements were determined. Serum 25-OHD was measured by immunoluminometric direct assay. Consumption of foods rich in vitamin D, sunscreen use and vitamin consumption were assessed through applied questionnaires. 62.1% of the subjects had insufficiency of 25-OHD (21-29 ng/ml) and 20.2% had deficiency (<20 ng/ml). Obese subjects (BMI >or=95th percentile for age and gender) had significantly lower concentration of 25-OHD than nonobese. Predictors of 25-OHD concentration were, in order of significance: percentage of body fat, BMI, triceps skin fold, and waist circumference (WC). A significantly higher rate of 25-OHD deficiency was observed in children with inadequate milk/yoghurt consumption, but no difference was found for other foods, physical activity (PA) or screen-time. In a multivariate model, being obese was significantly associated with the risk of 25-OHD deficiency, after adjustment for PA, screen-time, skin phototype, ingestion of milk/yoghurt, fish, cheese, and carbonated beverages. A high prevalence of vitamin D deficiency and an inverse association between serum 25-OHD concentration and obesity was found.

Role of CD4+CD25+highFoxp3+CD62L+ regulatory T cells and invariant NKT cells in human allogeneic hematopoietic stem cell transplantation.

Allogeneic hematopoietic stem cell transplantation (HSCT) is the treatment of choice for some hematological diseases; however, graft-versus-host disease (GVHD) is still one of the most important and deleterious complications. Regulatory T cells and iNKT cells can decrease the incidence and severity of GVHD, while preserving the graft-versus-tumor response. In order to analyze the relationship between the transfused dose of these cells, the presence of GVHD and survival, 15 normal donors and 15 patients with hematological diseases who underwent allogeneic HSCT from HLA-identical siblings were studied. The mobilization and infused doses of valpha24-vbeta11(iNKT cells) lymphocytes and CD4+CD25+FoxP3+, CD4+CD25+FoxP3+CD62L+, regulatory T cells were analyzed. All patients were conditioned with busulfan and cyclophosphamide and received cyclosporine and methotrexate as GVHD prophylaxis. iNKT and FoxP3 cells were mobilized after G-CSF administration. Acute GVHD was present in 9 of 15 (60%) and cGVHD in 7 of 13 (54%) patients. Patients who received a dose <0.6 x 10(6)/kg of iNKT cells and >4 x 10(6)/kg of FoxP3 had better disease-free survival and overall survival. Individuals transfused with >1.1 x 10(6)/kg of FoxP3+ CD62L+ Treg cells had better overall survival. In conclusion, iNKT and Treg cells are mobilized with G-CSF in healthy donors and the dose of iNKT cells and FoxP3 and CD62L+ regulatory T cells is of clinical importance in human HSCT.

Rapid complete response using intrathecal rituximab in a patient with leptomeningeal lymphomatosis due to mantle cell lymphoma.

Mantle cell lymphoma (MCL) is a B-cell lymphoid tumor that expresses CD20 and is associated with a poor prognosis. Central nervous system involvement has been associated with particularly dismal outcome. We report a 62-year-old male with MCL and meningeal lymphomatosis. The patient was treated with intrathecal rituximab (IT-R) 25 mg every third day for five doses with clearance of tumor after the third dose. Systemic therapy consisted of R-HyperCVAD alternating with rituximab, high-dose methotrexate, and cytarabine every 21 days, with IT-R on day 1 of each chemotherapy cycle. The patient was consolidated with an autologous stem cell transplant and remains in remission 23 months later. The use of IT-R and conventional intrathecal chemotherapy in MCLs is discussed here.

Trasplante autólogo de médula ósea como tratamiento de enfermedades autoinmunes: mecanismos y resultados / Autologous Bone Marrow Transplantation as a Treatment for Autoimmune Disease: Mechanisms and Results

Las enfermedades autoinmunes se caracterizan por una respuesta del sistema inmune del individuo hacia tejidos propios. Una línea de investigación actual es el tratamiento de estas enfermedades y el desarrollo de tolerancia. Una de las opciones en la búsqueda del desarrollo de tolerancia es el trasplante autólogo de médula ósea: la variantes del trasplante de médula ósea que hace uso de células progenitoras hematopoyéticas propias. La posibilidad de usar este tipo de trasplante como tratamiento de enfermedades autoinmunes se originó en los hallazgos de remisiones de enfermedades autoinmunes coexistentes, en pacientes que eran trasplantados por enfermedades oncológicas. En esta revisión presentemos el fundamento teórico de este tratamiento, así como una recopilación de los estudios preclínicos y clínicos más relevantes en esta materia. Aunque algún reporte inicial puso en duda la utilidad de dicho procedimiento, en general, los resultados son alentadores. Es necesario que más pacientes en diversos estadios de las enfermedades autoinmunes sean referidos a centros especializados de manera que sea posible recopilar la información de manera ordenada y sistemática, y se pueda arribar a un conocimiento sobre el papel que juega este tipo de tratamiento en las enfermedades autoinmunes.

Autoimmune diseases are characterized by immune response against self antigens. One of the current research interests in this field is oriented toward development of tolerance. One of the newest options in the search for tolerance is autologous bone marrow transpiantation: a variant of bone marrow transplant in which the patient's own hematopoietic stem cells are reinfused after myeloablative therapy. The idea of using bone marrow transplant in treatment of autoimmune diseases derived from observing remission in autoinmune diseases in patients transplanted due to coexisting neoplastic disease. Although an isolated initial report of bone marrow transplant as treatment for autoimmune disease questioned the utility of this procedure, over all, results are encouraging. To compile information in a programmed and systematic manner, it is necessary to send more patients in all stages of immune diseases to specialized centers to be included in large multicenter randomized trials. In time, the role for this procedure in autoimmune diseases will become clear.