In vivo murine model of acquired resistance in myeloma reveals differential mechanisms for lenalidomide and pomalidomide in combination with dexamethasone.

The development of resistance to therapy is unavoidable in the history of multiple myeloma patients. Therefore, the study of its characteristics and mechanisms is critical in the search for novel therapeutic approaches to overcome it. This effort is hampered by the absence of appropriate preclinical models, especially those mimicking acquired resistance. Here we present an in vivo model of acquired resistance based on the continuous treatment of mice bearing subcutaneous MM1S plasmacytomas. Xenografts acquired resistance to two generations of immunomodulatory drugs (IMiDs; lenalidomide and pomalidomide) in combination with dexamethasone, that was reversible after a wash-out period. Furthermore, lenalidomide-dexamethasone (LD) or pomalidomide-dexamethasone (PD) did not display cross-resistance, which could be due to the differential requirements of the key target Cereblon and its substrates Aiolos and Ikaros observed in cells resistant to each combination. Differential gene expression profiles of LD and PD could also explain the absence of cross-resistance. Onset of resistance to both combinations was accompanied by upregulation of the mitogen-activated protein kinase/extracellular signal-regulated kinase (ERK) kinase (MEK)/ERK pathway and addition of selumetinib, a small-molecule MEK inhibitor, could resensitize resistant cells. Our results provide insights into the mechanisms of acquired resistance to LD and PD combinations and offer possible therapeutic approaches to addressing IMiD resistance in the clinic.

The epoxyketone-based proteasome inhibitors carfilzomib and orally bioavailable oprozomib have anti-resorptive and bone-anabolic activity in addition to anti-myeloma effects.

Proteasome inhibitors (PIs), namely bortezomib, have become a cornerstone therapy for multiple myeloma (MM), potently reducing tumor burden and inhibiting pathologic bone destruction. In clinical trials, carfilzomib, a next generation epoxyketone-based irreversible PI, has exhibited potent anti-myeloma efficacy and decreased side effects compared with bortezomib. Carfilzomib and its orally bioavailable analog oprozomib, effectively decreased MM cell viability following continual or transient treatment mimicking in vivo pharmacokinetics. Interactions between myeloma cells and the bone marrow (BM) microenvironment augment the number and activity of bone-resorbing osteoclasts (OCs) while inhibiting bone-forming osteoblasts (OBs), resulting in increased tumor growth and osteolytic lesions. At clinically relevant concentrations, carfilzomib and oprozomib directly inhibited OC formation and bone resorption in vitro, while enhancing osteogenic differentiation and matrix mineralization. Accordingly, carfilzomib and oprozomib increased trabecular bone volume, decreased bone resorption and enhanced bone formation in non-tumor bearing mice. Finally, in mouse models of disseminated MM, the epoxyketone-based PIs decreased murine 5TGM1 and human RPMI-8226 tumor burden and prevented bone loss. These data demonstrate that, in addition to anti-myeloma properties, carfilzomib and oprozomib effectively shift the bone microenvironment from a catabolic to an anabolic state and, similar to bortezomib, may decrease skeletal complications of MM.

Mesenchymal stem cells from multiple myeloma patients display distinct genomic profile as compared with those from normal donors.

It is an open question whether in multiple myeloma (MM) bone marrow stromal cells contain genomic alterations, which may contribute to the pathogenesis of the disease. We conducted an array-based comparative genomic hybridization (array-CGH) analysis to compare the extent of unbalanced genomic alterations in mesenchymal stem cells from 21 myeloma patients (MM-MSCs) and 12 normal donors (ND-MSCs) after in vitro culture expansion. Whereas ND-MSCs were devoid of genomic imbalances, several non-recurrent chromosomal gains and losses (>1 Mb size) were detected in MM-MSCs. Using real-time reverse transcription PCR, we found correlative deregulated expression for five genes encoded in regions for which genomic imbalances were detected using array-CGH. In addition, only MM-MSCs showed a specific pattern of 'hot-spot' regions with discrete (<1 Mb) genomic alterations, some of which were confirmed using fluorescence in situ hybridization (FISH). Within MM-MSC samples, unsupervised cluster analysis did not correlate with particular clinicobiological features of MM patients. We also explored whether cytogenetic abnormalities present in myelomatous plasma cells (PCs) were shared by matching MSCs from the same patients using FISH. All MM-MSCs were cytogenetically normal for the tested genomic alterations. Therefore we cannot support a common progenitor for myeloma PCs and MSCs.

Grupo de apoyo y ayuda mutua a cuidadores informales de ancianos dependientes / Group of support and mutual help for informal carers of dependent elderly people

Poner en funcionamiento un grupo de apoyo y ayuda mutua para cuidadores informales de ancianos dependiente en el Centro de Salud “Vargas” es una necesidad que no se puede relegar por más tiempo.Apoyar a los cuidadores, escucharles, concienciarles de la importancia que tiene la tarea que realizan, tanto para el familiar que cuidan como para el resto de la sociedad, ayudarles y enseñarles a que se cuiden a sí mismos en este período, es el proyecto que queremos llevar a cabo, con la esperanza de que sea beneficioso tanto para el cuidador como para el anciano cuidado

Bringing into operation a group of support and mutual help for informal carers of dependent elderly people at “Vargas” Health Centre is a necessity that can´t be postponed any longer. The project which we want to carry out takes into account the welfare of both the carer and the elderly and consist of supporting the carers by listening to them, by raising their awareness about the importance of the work they do, both for the relative they look after and for society as a whole, and by teaching them to look after themselves during the caring period

Reflexiones sobre un programa de prácticas de Enfermería Geriátrica en, un centro de salud con perfil geriátrico / Reflexions about a Geriatric Nursing training program in a health centre with a geriatric profile

La oportunidad que nos ha brindado la Escuela de Enfermería de Cantabria de participar en la formación de los alumnos en el área de la Geriatría ha sido una experiencia muy positiva tanto para los alumnos, que se benefician de todo lo que la Atención Primaria puede aportar tanto en la Gerontología como en la Geriatría, como para las enfermeras de este centro de salud, que nos hemos concienciado de lo que significa trabajar en esta área. La formación de los alumnos en Geriatría es un reto para las enfermeras que tenemos la oportunidad de colaborar en ella y a la vez es una responsabilidad, ya que estos alumnos de hoy van a ser los profesionales que en el futuro atiendan a la población anciana de nuestra sociedad y por lo tanto, una buena formación se traduce, en la mayoría de los casos, en una atención de calidad para nuestros mayores y además en un beneficio para nuestra profesión, pues unos buenos profesionales dan calidad a la Enfermería (AU)

Composting of the solid fraction of olive mill wastewater with olive leaves: organic matter degradation and biological activity.

The flocculated solid fraction of olive mill wastewaters, obtained from two different olive oil extraction systems (FOMW1 and FOMW2) was composted, with olive leaves (OL) as bulking agent, by the static pile system (Rutgers). The dynamic of organic matter (OM) degradation during composting and its relationship with the basal respiration and fluorescein diacetate (FDA) hydrolytic activity, as indicators of biological activity, were studied. Two mixtures were prepared: C1, from 65% FOMW1 plus 35% OL; and C2, from 74% FOMW2 plus 25% OL and 1% urea. The biooxidative phase of composting in C1, which had a high initial C/N ratio, was long, leading to a high OM degradation, mainly of the lignocellulosic compounds. The water-soluble organic carbon content, C/N ratio and the urea supplied as a N source for the C2 compost make this mixture more adequate for composting, as it had a shorter composting time than C1, and developed a microbial population with a high metabolic activity. The results for basal respiration in C1 and C2 were correlated at a high probability level with those of FDA hydrolysis, and both parameters can be used for establishing the degree of biological stability of the composting material.

Growth of ornamental plants in two composts prepared from agroindustrial wastes.

Two composts prepared from agroindustrial wastes were assayed as substrates: C1 from brewing waste (yeast and malt) plus lemon tree prunings; and C2 from the solid fraction of olive mill wastewater plus olive leaves. Sixteen substrates were prepared by combining each compost with Sphagnum peat or a commercial substrate (CS) in different proportions. The nutrients (N and K) provided by the composts, which acted as slow-release fertilisers, influenced especially the development of calendula, although the physical and physico-chemical properties such as total pore space and electrical conductivity (EC) were also relevant. On the other hand, in the salt-sensitive calceolaria hybrid, EC and chloride concentration were the main factors influencing growth. Adequate substrates for the development of calendula can be prepared by mixing C1 at up to 75% with peat or at up to 50% with CS, and C2 at up to 50% with peat or CS. For calceolaria, the substrate should have a lower proportion of compost, C1 at up to 50% and C2 at up to 25%, both mixed with peat or CS. Therefore, composts of agroindustrial origin such as these can be used as an alternative to peat and CSs for growing ornamental plants. provided the mixture contains at least 25% peat or CS.

[Total plasma homocysteine levels. Relationship with plasmatic folic acid levels and 677C T polymorphism of 5,10-methylenetetrahydrofolate reductase]. / Concentración plasmática de homocisteína: relación con los niveles plasmáticos de ácido fólico y con el polimorfismo 677C T de la 5,10-metilenotetrahidrofolato reductasa.

BACKGROUND: Moderately increased plasma homocysteine (Hcy) in children has been associated with stroke and venous thrombosis and with a parental history of cardiovascular disease (CVD). Evaluation of Hcy concentrations during childhood and study of the factors determining its concentrations could play an important role in the primary prevention of CVD. Objective To detect cases of hyperhomocystinemia and to examine the association between Hcy levels and plasma folic acid levels and 677C T polymorphism of 5,10-methylenetetrahydrofolate reductase (MTHFR). METHODS: The relationship between plasma Hcy levels, plasma folic acid levels, and the three genotypes of 677C T MTHFR polymorphism was investigated in 127 children (aged 2-18 years) and in 105 parents by multiple linear regression. RESULTS: The median Hcy levels were 5.00 mol/l in the children and 8.00 mol/l in the parents. Plasma folic acid levels were normal in all of the patients. The prevalence of the three genotypes in the children was 32.3 % for the CC genotype, 42.5 % for the CT genotype and 15.7 % for the TT genotype. Hcy concentrations were significantly higher in children with the TT genotype (p 0.018). Multiple linear regression revealed a positive direct effect of age (b 0.029, p 0.002) and a negative effect of genotype TT (b 3.886, p 0.002) on Hcy concentration. Hcy concentration was inversely correlated with folic acid levels but this correlation did not reach statistical significance. CONCLUSIONS: No cases of hyperhomocystinemia were found. To evaluate Hcy, age and plasma folic acid levels have to be taken into account in case there is a 677C T mutation. Hcy concentrations should be determined in older children with a family history of atherothrombosis and other risk factors for premature CVD.

Concentración plasmática de homocisteína: relación con los niveles plasmáticos de ácido fólico y con el polimorfismo 677 ct de la 5,10-metilenotetrahidrofolato reductasa / Total plasma homocysteine levels. Relationship with plasmatic folic acid levels and 677 CT polymorphism of 5,10-methylenetetrahydrofolate reductase

Antecedentes: El aumento moderado de la homocisteína plasmática en niños se ha relacionado con infartos cerebrales y trombosis venosas y con los antecedentes familiares de enfermedad coronaria prematura (ECP). La determinación de homocisteína en la infancia y el estudio de los factores que determinan su concentración podría ser importante para la prevención primaria de la ECP. Objetivo: Detectar algún caso de hiperhomocistinemia y valorar su relación con la concentración plasmática de ácido fólico y el polimorfismo 677C T de la 5,10-metilenotetrahidrofolato reductasa (MTHFR). Métodos: Se ha estudiado mediante la regresión lineal múltiple la relación entre la concentración plasmática de homocisteína, la del ácido fólico y los tres genotipos de la mutación 677C T de la MTHFR en 127 niños de entre 2 y 18 años y 105 de sus progenitores. Resultados: La concentración de homocisteína (mediana) fue de 5,00 y 8,00 mol/l en los niños y sus progenitores, respectivamente. Los valores plasmáticos de ácido fólico se encontraban todos en el rango de la normalidad. La prevalencia de los tres genotipos en los niños fue de 32,3% para el genotipo CC, 42,5% para el CT y 15,7% para el TT. La concentración de homocisteína era significativamente mayor con el genotipo TT (p = 0,018). En la regresión lineal múltiple se encontró un efecto directo positivo de la edad (b = 0,029; p = 0,001) y negativo del genotipo TT (b = -3,886; p = 0,002) sobre la concentración de homocisteína. El coeficiente de regresión de la concentración de ácido fólico aunque de signo negativo, no alcanzó significación estadística. Conclusiones: No se ha encontrado ningún caso de hiperhomocistinemia. Al valorar la homocisteína hay que tener en cuenta la edad y en caso de existir la mutación 677C T, los valores plasmáticos de ácido fólico. Sería conveniente determinar la homocisteína en los niños de mayor edad con antecedentes familiares de aterotrombosis y con otros factores de riesgo para la ECP (AU)

HLA genes in Arabic-speaking Moroccans: close relatedness to Berbers and Iberians.

The gene profile of Arabic-speaking Moroccans has been compared with those of other Mediterranean populations in order to provide additional information about the history of their origins. Our HLA data suggest that most Moroccans are of a Berber (Imazighen) origin and that Arabs who invaded North Africa and Spain in the 7th century A.D. did not substantially contributed to the gene pool; however, they imposed their advanced culture and their religion. Present-day Egyptians are also related to Moroccan Berbers and this supports an ancient Saharan origin for part of the present-day Mediterraneans, particularly for the Arabic-speaking ones (also Algerians) and also for the older substratum of Mediterranean people.

Pilot screening programme for cystinuria in the Valencian community.

Cystinuria is an autosomal recessive disorder of the kidneys and small intestine, affecting a luminal transport mechanism shared by cystine, ornithine, arginine and lysine. When cystine exceeds its solubility at low pH, the risk of stone formation increases. The data reported in the literature show a variation for the incidence of cystinuria, from 1 in 600 to 1 in 17,000, depending on the definition of cystinuria and the method used for screening the population. We set up a pilot screening programme to determine the incidence of cystinuria in the population of the Valencian Community. Urine filter paper samples submitted for the neonatal screening programme from 33,995 newborns (5-10 days old) were used for the study. Thin layer chromatography (TLC) was performed to screen cystinuric patients. To confirm positive filter paper samples, liquid samples were requested and TLC as well as the cyanide-nitroprusside test (CNT) were performed. Final diagnosis was achieved by quantifying cystine, lysine, ornithine and arginine using high-performance liquid chromatography (HPLC) in children's urine samples which remained positive for TLC and CNT for more than 1 year. We conclude that the incidence of subjects at risk for cystine stones in the Valencian Community is 1:1887. TLC is shown as a reliable method to perform newborn screening in large population to detect cystinuric subjects. Additional studies, including characterization of appropriate haplotypes, should be carried out for a more precise identification of the frequency of the different types of cystinuria in our population.

[Homocystinuria: effectiveness of the treatment with pyridoxine, folic acid, and betaine]. / Homocistinuria: eficacia del tratamiento con piridoxina, ácido fólico y betaína.

We present the results achieved with vitamin (pyridoxine and folic acid) and betaine (trimethyl-glycine) treatment of three patients with homocystinuria. Cases 1 and 2 were detected by having clinical findings suggestive of the disease (ocular and orthopedic alterations) and case 3 was diagnosed after a family metabolic screening was done. All presented a positive Brand's test and an abnormal elevation of plasma and urine homocysteine, as well as high methionine and low cystine levels in the plasma. Initially, when pyridoxine (600 mg/d) and folic acid (10 mg/d) were given for one month, a partial fall in the homocysteine levels was observed in cases 2 and 3, but not in case 1. When betaine was added (6 g/d), homocysteine disappeared from the plasma after the first month in cases 2 and 3, but only after the third month in case 1. Case 1 also showed a moderate clinical improvement in behavior and school performance. The treatment was maintained for two years in case 1, and for one year in cases 2 and 3. After betaine therapy, no disturbances were observed in the hepatic, renal and bone marrow functions, nor were there any clinically relevant ill-effects. These findings show that betaine offers a therapeutic alternative in the treatment of this disease, independent of the patient's response to pyridoxine.

[Circulating immune complexes in the serum of tumor patients]. / Complejos inmunes circulantes en el suero de pacientes con tumores.

The conditions for the detection of circulating immune complexes (CIC) were established by combination of polyethylene glycol precipitation with a nephelometric determination on the precipitate of endogenous C1q. The CIC precipitated from the sera of patients with breast carcinoma, malignant melanoma or autoimmune diseases were dissociated in SDS-beta-mercaptoethanol buffer and no differences were observed in the electrophoretic patterns os polyacrylamide gels allowing to characterize tumour specific antigens.