Minimal Residual Disease Monitoring with Next-Generation Sequencing Methodologies in Hematological Malignancies

Ultra-deep next-generation sequencing has emerged in recent years as an important diagnostic tool for the detection and follow-up of tumor burden in most of the known hematopoietic malignancies. Meticulous and high-throughput methods for the lowest possible quantified disease are needed to address the deficiencies of more classical techniques. Precision-based approaches will allow us to correctly stratify each patient based on the minimal residual disease (MRD) after a treatment cycle. In this review, we consider the most prominent ways to approach next-generation sequencing methodologies to follow-up MRD in hematological neoplasms.This work received no specific external funding; our research group was supported by the Fundación Cris contra el cáncer.S

Ocena minimalnej choroby resztkowej w szpiczaku plazmocytowym

Minimal residual disease (MRD) in a patient with multiple myeloma (MM) is defined as the mini-mum levels of pathological plasma cells remaining after treatment when a patient is in complete response (CR). The ultimate aim of studying MRD is to identify patients with different prognosis and to tailor treatment for individual patients. MRD studies in MM should be recommended in young patients in CR after autologous hematopoietic stem cells transplantation and in older patients in CR after regimens including proteasome inhibitors. Bone marrow is the only recommend location to assess MRD in MM. The recommended methods of MRD testing include next generation sequencing of immunoglobulin genes or multiparametric flow cytometry (MFC), depending on the experience of each center and the possibility of study samples being available in the first 24 hours for MFC analysis. MRD should be considered as a therapeutic objective. However, there is not enough evidence for taking clinical decisions based on MRD status alone, and for this reason we encourage the design of new clinical studies to address these questions.Minimalną chorobę resztkową (MRD) u pacjenta z rozpoznaniem szpiczaka plazmocytowego defi-niuje się jako populację nowotworowych komórek plazmatycznych, która pozostała w organizmie chorego po osiągnięciu odpowiedzi całkowitej (CR). Ostatecznym celem badań MRD jest dążenie do identyfikacji chorych o odmiennym rokowaniu i indywidualizacji leczenia na tej podstawie. Ocenę MRD u chorych na szpiczaka plazmocytowego zaleca się u młodszych pacjentów, którzy osiągną CR po przeszczepieniu autologicznych krwiotwórczych komórek macierzystych oraz u chorych starszych osiągających CR po chemioterapii opartej na inhibitorach proteasomu. Powinno się oznaczać MRD wyłącznie w szpiku kostnym. Do rekomendowanych metod oceny MRD w szpiczaku plazmocytowym zalicza się sekwencjonowanie następnej generacji genów immunoglobulinowych oraz wieloparametryczną cytometrię przepływową, przy czym wybór jednej z tych metod powinien zależeć od doświadczenia danego ośrodka oraz możliwości wykonania badania cytometrycznego w czasie 24 godzin od pobrania próbki szpiku. Eradykację MRD powinna się obecnie uważać za istotny cel terapii szpiczaka plazmocytowego. Jednak, ze względu na brak wystarczających danych do podejmowania decyzji klinicznych wyłącznie na podstawie wyniku badania MRD, istnieje po¬trzeba dalszych, dobrze zaprojektowanych badań klinicznych w tym zakresie

Conversaciones con María José Hidalgo de la Vega: en torno al poder, los poderes femeninos y las mujeres poderosas del mundo antiguo

Entre quienes son especialistas de la Historia Antigua y de los estudios his- tóricos de las mujeres de laAntigüedad, el nombre de María José Hidalgo de la Vega evoca siempre a una pionera, por haber sido una de lasprimeras catedráticas de esta especialidad en la Universidad española, por su compromiso con el femi- nismo y por sus trabajos sobre las mujeres poderosas del Imperio romano, como refleja su biografía sobre Zenobia, reina de Palmira, de reciente publicación. Su trayectoria académica y personal representa, sin duda, a toda una generación de historiadoras que se formaron en la España tardo-franquista, en un contexto de profundastransformaciones sociales y políticas determinantes a la hora de plantear el estudio del pasado desde el compromisosocial con el presente y con las mujeres. Nacida en Málaga, María José Hidalgo de la Vega se trasladó a Granada para cursar sus estudios de licenciatura en Filología Clásica, fue alumna y discípula de Marcelo Vigil, prestigioso profesor, que trajo a la España de los años sesenta y setenta las visiones propias de la HistoriaSocial, totalmente novedosas en ese momento. Fallecido prematuramente, fue su marido y padre de su única hija, Lucía, y marcó indudablemente su interés por las sociedades del Mediterráneo antiguo y la literatura latina. Muy pronto, sin haber cumplido losveinticinco años, abandonó tierras andaluzas para fijar su residencia en Salamanca, dónde se jubiló tras una intensa biografía académica. De su implicación en la vida de la Univer- sidad, ha de señalarse que fue vicerrectora, decana, directora de Departamento y de la revista, Studia Historica. Historia Antigua, además de participar en otras múltiples actividades. Era importante esta dedicación en momentos de cambios y transformaciones de la Universidad española en la etapa de la transición. A la vez, desarrolló una brillante carrerainvestigadora, como revelan sus numerosas publi- caciones, que continúan en la actualidad, claramenterelacionadas con la Historia Social, pero en las que poco a poco ha ido incorporando las aportaciones de los estudios culturales y siempre ligadas a sus indagaciones sobre el poder, la política y la religión, dónde fue incorporando las mujeres y el género. La coordinación del grupo de investigación Epirus es también una clara muestra de su capacidad para organizar equipos y proyectos, facilitando el trabajo de los colegas cercanos, sus discípulos y discípulas

Medical Laboratory Accreditation according to ISO 15189:2003.The Mexican Experience

Međunarodna norma ISO15189:2003 iziskuje od laboratorija da udovolje zahtjevima za upravljanje kvalitetom kao i tehničkim uvjetima, uključujući prije- i poslije-analitičke faze te sam analitički proces. Norma obuhvaća i teme kao što su sigurnost i etika u medicinskom laboratoriju. S obzirom da se primjenjuju lokalni, nacionalni i regionalni propisi, ISO 15189:2003 traži da medicinski laboratoriji dokažu svoju kompetentnost neovisnom procjenitelju, a to je obično nacionalno akreditacijsko tijelo. U Meksiku je vlada imenovala Meksički akreditacijski entitet (EMA) za akreditiranje medicinskih laboratorija u skladu s ISO15189:2003; ta organizacija djeluje usko surađujući s Meksičkim društvom za kliničku biokemiju (AMBC).International standard ISO15189:2003 requires that laboratories comply with requirements for quality management and technical requirements, including pre-and post-analytical phases, as well as the analytical process itself. Topics such as laboratory safety and medical laboratory ethics are also included. As local, national and regional regulations may apply, ISO15189:2003 requires that medical laboratories demonstrate their competence to a third party assessor, which is usually a national accreditation body. In Mexico, the government has appointed the Mexican Accreditation Entity (EMA), to accredit medical laboratories in accordance to ISO 15189:2003; this organization works in close cooperation with the Mexican Association of Clinical Biochemistry (AMBC)

Procesos de enseñanza/aprendizaje entre pares al interior del second life

Este artículo presenta el análisis de algunas prácticas no institucionalizadas de enseñanza-aprendizaje al interior del Second Life, a través de las cuales se visibilizan estrategias mediacionales como la descripción, modelado y valoración de procedimientos. Una particularidad central de las prácticas seleccionadas para este estudio, es que los usuarios participantes pese a que adoptan roles de profesores/tutores o aprendices, no se asumen como tales. Metodológicamente, el acercamiento es realizado a través de observación participativa en línea dentro de la Isla Reforma en el Second Life, así como de entrevistas semiestructuradas. Entre los hallazgos destacan: a) La repetición en el espacio virtual de procesos de enseñanza interiorizados por los sujetos en sus experiencias escolarizadas y analógicas previas. b) El grado de “libertad y flexibilidad” que le confieren algunos usuarios a las interacciones en el Second Life VS las que estarían dispuestos a entablar en una relación cara a cara. c) La relación entre apoyo/tutoría/donación y el posicionamiento del sujeto a través de su avatar, en la grupalidad. d) El potencial que la enseñanza/aprendizaje entre pares representa para procesos de formación continua y no institucionalizada. Palabras clave Enseñanza/aprendizaje entre pares, trabajo colaborativo, comunidad virtual, Second Life

Effect of n-3 polyunsaturated fatty acid supplementation on metabolic and inflammatory biomarkers in type 2 diabetes mellitus patients

Artículo derivado de un proyecto de investigación de Suplementación con ácidos grasos poliinsaturados n-3 y vitamina D en pacientes con diabetes mellitus tipo 2Abstract: Background: Type 2 diabetes mellitus (T2DM) is accompanied by chronic low-grade inflammation, with an imbalance in the secretion of adipokines and, worsening insulin resistance. Supplementation with n-3 PUFA in T2DM decreases inflammatory markers, the purpose of the study was to investigate the effect of n-3 PUFA supplementation on adipokines, metabolic control, and lipid profile in T2DM Mexican adults. Methods: In a randomized, single-blind, placebo-controlled pilot study, 54 patients with T2DM received 520 mg of DHA + EPA-enriched fish-oil (FOG) or a placebo (PG) daily. Baseline and 24-week anthropometric and biochemical measurements included glucose, insulin, glycosylated hemoglobin (Hb1Ac), leptin, adiponectin, resistin, and lipid profile; n-3 PUFA intake was calculated in g/day. Results: Waist circumference and blood glucose showed significant reductions in the FOG group (p = 0.001 and p = 0.011, respectively). Hb1Ac (p = 0.009 and p = 0.004), leptin (p < 0.000 and p < 0.000), and leptin/adiponectin ratio (p < 0.000 and p < 0.000) decreased significantly in both groups after 24 weeks (FOG and PG respectively). Serum resistin (FOG p < 0.000 and PG p = 0.001), insulin (FOG p < 0.000 and PG p < 0.000), and HOMA-IR (FOG p = 0.000 and PG p < 0.000) increased significantly in both groups. FOG had an overall improvement in the lipid profile with a significant decrease in triacylgycerols (p = 0.002) and atherogenic index (p = 0.031); in contrast, the PG group had increased total cholesterol (p < 0.000), non-HDL cholesterol (p < 0.000), and atherogenic index (p = 0.017). Conclusions: We found a beneficial effect of n-3 PUFA supplementation on waist circumference, glucose, Hb1Ac, leptin, leptin/adiponectin ratio, and lipid profile, without significant changes in adiponectin, and increases in resistin, insulin, and HOMA-IR in both groups.CONACyT, Desarrollo Científico para atender problemas nacionales, No. 21294

Current challenges in chronic bronchial infection in patients with chronic obstructive pulmonary disease

Currently, chronic obstructive pulmonary disease (COPD) patients and their physicians face a number of significant clinical challenges, one of which is the high degree of uncertainty related to chronic bronchial infection (CBI). By reviewing the current literature, several challenges can be identified, which should be considered as goals for research. One of these is to establish the bases for identifying the biological and clinical implications of the presence of potentially pathogenic microorganisms in the airways that should be more clearly elucidated according to the COPD phenotype. Another urgent area of research is the role of long-term preventive antibiotics. Clinical trials need to be carried out with inhaled antibiotic therapy to help clarify the profile of those antibiotics. The role of inhaled corticosteroids in patients with COPD and CBI needs to be studied to instruct the clinical management of these patients. Finally, it should be explored and confirmed whether a suitable antimicrobial treatment during exacerbations may contribute to breaking the vicious circle of CBI in COPD. The present review addresses the current state of the art in these areas to provide evidence which will enable us to progressively plan better healthcare for these patients

Therapeutic effects of telomerase in mice with pulmonary fibrosis induced by damage to the lungs and short telomeres

Pulmonary fibrosis is a fatal lung disease characterized by fibrotic foci and inflammatory infiltrates. Short telomeres can impair tissue regeneration and are found both in hereditary and sporadic cases. We show here that telomerase expression using AAV9 vectors shows therapeutic effects in a mouse model of pulmonary fibrosis owing to a low-dose bleomycin insult and short telomeres. AAV9 preferentially targets regenerative alveolar type II cells (ATII). AAV9-Tert-treated mice show improved lung function and lower inflammation and fibrosis at 1-3 weeks after viral treatment, and improvement or disappearance of the fibrosis at 8 weeks after treatment. AAV9-Tert treatment leads to longer telomeres and increased proliferation of ATII cells, as well as lower DNA damage, apoptosis, and senescence. Transcriptome analysis of ATII cells confirms downregulation of fibrosis and inflammation pathways. We provide a proof-of-principle that telomerase activation may represent an effective treatment for pulmonary fibrosis provoked or associated with short telomeres.We are indebted to D Megias for microscopy analysis, to J Mun˜ oz and F Garcı´a for hydroxiproline analysis as well as to CNIO Histopathological Unit. The research was funded by project SAF2013- 45111-R of Societal Changes Programme of the Spanish Ministry of Economics and Competitiveness (MINECO) co-financed through the European Fund of Regional Development (FEDER), Fundacio´n Botı´n and Banco Santander (Santander Universities Global Division) and Roche Extending the Innova- tion Network Program (EIN) Academia Partnering Programme.S

Versatile Graphene-Based Platform for Robust Nanobiohybrid Interfaces

Technologically useful and robust graphene-based interfaces for devices require the introduction of highly selective, stable, and covalently bonded functionalities on the graphene surface, whilst essentially retaining the electronic properties of the pristine layer. This work demonstrates that highly controlled, ultrahigh vacuum covalent chemical functionalization of graphene sheets with a thiol-terminated molecule provides a robust and tunable platform for the development of hybrid nanostructures in different environments. We employ this facile strategy to covalently couple two representative systems of broad interest: metal nanoparticles, via S-metal bonds, and thiol-modified DNA aptamers, via disulfide bridges. Both systems, which have been characterized by a multi-technique approach, remain firmly anchored to the graphene surface even after several washing cycles. Atomic force microscopy images demonstrate that the conjugated aptamer retains the functionality required to recognize a target protein. This methodology opens a new route to the integration of high-quality graphene layers into diverse technological platforms, including plasmonics, optoelectronics, or biosensing. With respect to the latter, the viability of a thiol-functionalized chemical vapor deposition graphene-based solution-gated field-effect transistor array was assessed

Ruxolitinib in combination with prednisone and nilotinib exhibit synergistic effects in human cells lines and primary cells from myeloproliferative neoplasms

Ruxolitinib is the front-line non-palliative treatment for myelofibrosis (MF). However, a significant number of patients lose or present suboptimal response, are resistant or have unacceptable toxicity. In an attempt to improve response and avoid the adverse effects of this drug, we evaluated the combination of 17 drugs with ruxolitinib in ex vivo models of peripheral blood mononuclear cells from MF patients and cell lines. We found that the combination ruxolitinib and nilotinib had a synergistic effect against MF cells (ΔEC50 nilotinib, -21.6%). Moreover, the addition of prednisone to combined ruxolitinib/nilotinib improved the synergistic effect in all MF samples studied. We evaluated the molecular mechanisms of combined ruxolitinib/nilotinib/prednisone and observed inhibition of JAK/STAT (STAT5, 69.2+11.8% inhibition) and MAPK (ERK, 29.4+4.5% inhibition) signaling pathways. Furthermore, we found that the triple therapy combination inhibited collagen protein and COL1A1 gene expression in human bone marrow mesenchymal cells. Taken together, we provide evidence that combined ruxolitinib/nilotinib/prednisone is a potential therapy for MF, possibly through the anti-fibrotic effect of nilotinib, the immunomodulatory effect of ruxolitinib and prednisone, and the anti-proliferative effect of ruxolitinib. This combination will be further investigated in a phase Ib/II clinical trial in MF.This study was supported by the Subdireccion General de Investigacion Sanitaria (Instituto de Salud Carlos III, Spain) grants PI13/02387 and PI16/01530, and the CRIS against Cancer foundation grant 2014/0120. M.L. holds a postdoctoral fellowship of the Spanish Ministry of Economy and Competitiveness (FPDI-2013-16409).S