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© 2019 Ferrata Storti Foundation., B-cell acute lymphoblastic leukemia is the commonest childhood cancer. In infants, B-cell acute lymphoblastic leukemia remains fatal, especially in patients with t(4;11), present in ~80% of cases. The pathogenesis of t(4;11)/KMT2A-AFF1+ (MLL-AF4+) infant B-cell acute lymphoblastic leukemia remains difficult to model, and the pathogenic contribution in cancer of the reciprocal fusions resulting from derivative translocated-chromosomes remains obscure. Here, “multi-layered” genome-wide analyses and validation were performed on a total of 124 de novo cases of infant B-cell acute lymphoblastic leukemia uniformly diagnosed and treated according to the Interfant 99/06 protocol. These patients showed the most silent mutational landscape reported so far for any sequenced pediatric cancer. Recurrent mutations were exclusively found in K-RAS and N-RAS, were subclonal and were frequently lost at relapse, despite a larger number of non-recurrent/non-silent mutations. Unlike non-MLL-rearranged B-cell acute lymphoblastic leukemias, B-cell receptor repertoire analysis revealed minor, non-expanded B-cell clones in t(4;11)+ infant B-cell acute lymphoblastic leukemia, and RNA-sequencing showed transcriptomic similarities between t(4;11)+ infant B-cell acute lymphoblastic leukemias and the most immature human fetal liver hematopoietic stem and progenitor cells, confirming a “pre-VDJ” fetal cellular origin for both t(4;11) and RASmut. The reciprocal fusion AF4-MLL was expressed in only 45% (19/43) of the t(4;11)+ patients, and HOXA cluster genes are exclusively expressed in AF4-MLL-expressing patients. Importantly, AF4-MLL/HOXA-expressing patients had a significantly better 4-year event-free survival (62.4% vs. 11.7%, P=0.001), and overall survival (73.7 vs. 25.2%, P=0.016). AF4-MLL expression retained its prognostic significance when analyzed in a Cox model adjusting for risk stratification according to the Interfant-06 protocol based on age at diagnosis, white blood cell count and response to prednisone. This study has clinical implications for disease outcome and diagnostic risk-stratification of t(4;11)+ infant B-cell acute lymphoblastic leukemia., This work was supported by the European Research Council (CoG-2014- 646903 to PM; and StG-2014-637904 to IV), the Spanish Ministry of Economy and Competitiveness (SAF-SAF2013- 43065 to PM and SAF2016-76758-R to IV), the Asociación Española Contra el Cáncer (AECC-CI-2015), FERO Foundation, and the ISCIII (PI14-01191) to CB. PM/IV also acknowledge financial support from The Obra Social La CaixaFundaciò Josep Carreras, The Inocente Inocente Foundation, Fundación Ramón Areces and The Generalitat de Catalunya (SGR330). IR was supported by a Programme Grant from Bloodwise (LLR 13001) and by the Oxford NIHR Biomedical Centre based at Oxford University Hospitals NHS Trust and University of Oxford. PM is an investigator of the Spanish Cell Therapy cooperative network (TERCEL). AR was supported by a Clinician Scientist Fellowship from Bloodwise (14041).

OBJECTIVE To assess whether apparently healthy subjects with a family history of systemic hypertension have a higher risk of presenting the insulin resistance syndrome. SUBJECTS Three hundred and eighty-six subjects aged 20-65 years. SETTING A middle socio-economic class urban community from Mexico City. METHOD All subjects and, when necessary, their first-degree relatives, answered a questionnaire and underwent a physical examination with measurement of height, weight and blood pressure. Serum insulin, glucose, cholesterol and triglycerides were measured during fasting and 2 h after an oral load of 75 g glucose. RESULTS A family history of systemic hypertension was present for 167 (43%) of the subjects, of whom 123 (31%) were obese. Subjects with a family history of hypertension had higher systolic blood pressures than did those without such a history (120 +/- 15 versus 115 +/- 10 mmHg). In the logistic regression model, the body mass index and age showed statistically significant effects on the fasting glucose:insulin ratio and on serum insulin levels after an oral load of glucose. When men and women were analysed separately, only in men were higher systolic and mean blood pressures and lower glucose:insulin ratios observed. In the logistic regression analysis the body mass index was a significant predictor of the glucose:insulin ratio and serum insulin levels after an oral load of glucose, especially in men. CONCLUSION Apparently healthy male offspring of hypertensive parents have higher blood pressure levels and lower insulin sensitivities than do offspring of normotensive parents. Insulin resistance was related to obesity, but not to a family history of hypertension, as had previously been reported by other research groups.

CD163 is a member of the family of proteins with scavenger receptor cysteine-rich domains, whose expression is restricted to monocytes and macrophages. It functions as a receptor for haemoglobin/haptoglobin complexes and it has also been implicated in the regulation of inflammatory processes. Treatment of monocytes/macrophages with phorbol esters, LPS or cross-linking of Fc gamma receptors, causes shedding of CD163 from cell surface by a protease-dependent mechanism. The level of soluble CD163 (sCD163) in serum and other body fluids has been considered a useful marker of macrophage activation. In addition, porcine CD163 has been shown to play a role in the infection of monocytes/macrophages by the African swine fever virus. The porcine CD163 cDNA has been cloned, and expressed in transfected CHO cells. Clones of CHO cells stably expressing porcine CD163 have been used for the characterization of three new mAbs against porcine CD163. Using two of these mAbs, that bind to different epitopes on CD163 molecule, a sandwich enzyme-linked immunoassay (ELISA) has been developed to measure levels of sCD163 in porcine sera and biological fluids. The assay was calibrated using lysates of CD163 transfectants, showing a sensitivity of 10E5 cells/mL, that allowed to detect sCD163 in sera and in culture supernatants of activated alveolar macrophages. This assay can be a useful method of monitoring the degree of activation of macrophages in a variety of inflammatory and infectious diseases of swine., CD163 es un miembro de la familia de proteínas con dominios de receptores "scavenger" ricos en cisteína, con una expresión restringida a células del linaje monocítico. Se ha demostrado su función como receptor de complejos hemoglobina/ haptoglobina, y se le ha implicado en la regulación de procesos inflamatorios. El tratamiento de monocitos/ macrófagos con ésteres de forbol, LPS, o mediante entrecruzamiento de receptores Fc gamma, provoca la liberación de CD163 de la superficie celular por un mecanismo dependiente de proteasas. La cantidad de CD163 soluble (sCD163) en suero y otros fluidos corporales se considera un indicador de la activación de los macrófagos. Recientemente, nuestro grupo demostró que CD163 está implicado en la infección de monocitos/macrófagos porcinos por el virus de la peste porcina africana. En este trabajo se describe la clonación del cDNA de CD163 porcino, y su expresión en células CHO transfectadas. Se obtuvieron clones de células CHO transfectadas que expresan de forma estable la molécula CD163 porcina, y se utilizaron para caracterizar tres nuevos anticuerpos monoclonales que reconocen específicamente CD163 porcino. El uso de dos de estos anticuerpos, que reconocen epítopos diferentes en CD163 porcino, ha permitido desarrollar un ensayo inmunoenzimático (ELISA) para medir la cantidad de sCD163. El ensayo se calibró con lisados de las células transfectadas que expresan CD163 porcino, mostrando una sensibilidad equivalente a 10E5 células/mL, suficiente para detectar la presencia de sCD163 en suero y sobrenadantes de cultivo de macrófagos alveolares activados. Este ensayo será de utilidad para determinar el grado de activación de los macrófagos porcinos en procesos inflamatorios y enfermedades infecciosas porcinas.

A cDNA containing the porcine Toll-like receptor 4 (TLR4) coding sequence has been cloned by RT-PCR from alveolar macrophages mRNA, and its complete sequence has been determined. The predicted amino acid sequence comprises an extracellular domain with 21 leucine-rich repeats (LRR) and a LRR-C-terminal (LRR-CT) motif, followed by a 30 amino acid transmembrane segment, and a 179 amino acid intracytoplasmic region containing the Toll/IL-1R domain. Pig TLR4 shows 63-80% amino acid sequence identity with those of cow, horse, cat, human, rabbit and mouse. The degree of sequence identity rises to over 90% in the TIR domain. The whole TLR4 sequence and its ectodomain were expressed as GFP fusion proteins in CHO cells. Using RT-PCR analysis, porcine TLR4 transcripts were detected in DCs, monocytes and macrophages, and in tissue samples of bone marrow, thymus, lymph node, spleen, brain, liver, kidney and ovary. The expressed protein will be used for the development of reagents. Knowledge of TLR4 expression will help to address mechanisms of immune induction by antigens and vaccines. © 2005 Elsevier Ltd. All rights reserved.

The influence that water-repellent products can have on physical and micro-structural properties of lime mortars, and lime plus pozzolan mortars has been studied. Three water repellent products have been used. Mixes of the previously mentioned three water repellents plus a biocide product were also applied. Treatments make the total porosity and saturation coefficient of both mortars to decrease, while colorimetric coordinates bear little alteration. All treatments with water repellent products provided mortars with a hydrophobic property index close to 100%. Durability of such mortars has been also studied: salt crystallization test, frost-thaw and dry-wet cycles, as well as ultraviolet radiation test were carried out. Relationship between mortars behavior and their porosity and saturation coefficient were found., En el presente trabajo se ha estudiado la influencia de la aplicación de productos hidrofugantes a morteros de cal y morteros de cal y puzolana sobre sus propiedades físicas y microestructurales. Se han estudiado tres productos hidrofugantes. También han sido estudiados dichos productos junto con un biocida. La porosidad total y el coeficiente de saturación de ambos tipos de morteros se ve reducido por el efecto de los tratamientos mientras que las coordenadas colorimétricas se ven poco alteradas. Todos los tratamientos confieren un índice de hidrofobicidad a los morteros próximo al 100%. Asimismo, también se ha estudiado la durabilidad de dichos morteros frente a la cristalización de sales, hielo-deshielo, los ciclos de humedad-sequedad y radiaciones ultravioleta. Se relaciona el comportamiento de los morteros con su porosidad y el coeficiente de saturación.

Amongst the most important signal transduction molecules involved in regulating growth and differentiation are the protein tyrosine kinases (PTK). Since T cell development is a consequence of interactions between thymic stromal cells (TSC) and thymocytes, identification of the PTK in both compartments is required to dissect the mechanisms that control this process. Here we report a search for PTK in mouse TSC, using RT-PCR to survey the repertoire of PTK mRNAs expressed in a freshly isolated TSC preparation. We identified 10 different PTK cDNAs among the 216 cDNAs sequenced, and demonstrate that transcripts of three of those (ufo, fyn and fer) are widely expressed among a large panel of immortalized thymic epithelial cell lines (TEC) and in primary cultures of TSC. Of the other seven, none were expressed in established TEC lines but, instead, displayed distinct expression patterns in cell types likely to have contaminated the fresh TSC preparation, i.e., macrophages, B cells, T cells and fibroblasts. Among the three PTK expressed in TEC lines, only one, ufo, exhibited expression exclusively in cells of non-hemopoietic origin. Although expression of ufo (also known as tyro 7, axl or ark) is not thymic-specific, in that it is also expressed in cell types of mesodermal origin in other tissues, its presence in TEC suggests a role for ufo in differentiation of the TSC compartment. Consistent with this notion, high-level expression of this receptor PTK at the protein level could be documented in every TEC line investigated, as well as in fresh thymus tissue sections. These data provide the first example of a receptor PTK in TSC and open new approaches to study the regulation of TSC differentiation