Effect of osteoprotegerin and Dickkopf-related protein 1 on radiological progression in tightly controlled rheumatoid arthritis.

OBJECTIVE: To analyze the association between circulating osteoprotegerin (OPG) and Dickkopf-related protein 1 (DKK-1) and radiological progression in patients with tightly controlled rheumatoid arthritis (RA). METHODS: Serum levels of OPG and DKK-1 were measured in 97 RA patients who were treated according to a treat-to-target strategy (T2T) aimed at remission (DAS28<2.6). Radiologic joint damage progression was assessed by changes in the total Sharp-van der Heijde score (SHS) on serial radiographs of the hands and feet. The independent association between these biomarker levels and the structural damage endpoint was examined using regression analysis. RESULTS: The mean age of the 97 RA patients (68 women) at the time of the study was 54 ± 14 years, and the median disease duration was 1.6 ± 1.5 years. Most patients were seropositive for either RF or ACPA, and the large majority (76%) were in remission or had low disease activity. After a median follow-up time of 3.3 ± 1.5 years (range, 1-7.5 yrs.), the mean total SHS annual progression was 0.88 ± 2.20 units. Fifty-two percent of the patients had no progression (defined as a total SHS of zero). The mean serum OPG level did not change significantly over the study period (from 3.9 ± 1.8 to 4.07 ± 2.23 pmol/L), whereas the mean serum DKK-1 level decreased, although not significantly (from 29.9 ± 10.9 to 23.6 ± 18.8 pmol/L). In the multivariate analysis, the predictive factors increasing the likelihood of total SHS progression were age (OR per year = 1.10; p = 0.003) and a high mean C-reactive protein level over the study period (OR = 1.29; p = 0.005). Circulating OPG showed a protective effect reducing the likelihood of joint space narrowing by 60% (95% CI: 0.38-0.94) and the total SHS progression by 48% (95% CI: 0.28-0.83). The DKK-1 levels were not associated with radiological progression. CONCLUSION: In patients with tightly controlled RA, serum OPG was inversely associated with progression of joint destruction. This biomarker may be useful in combination with other risk factors to improve prediction in patients in clinical remission or low disease activity state

Novel intragenic deletion within the FXN gene in a patient with typical phenotype of Friedreich ataxia: may be more prevalent than we think?

Background Friedreich ataxia is the most common inherited ataxia in Europe and is mainly caused by biallelic pathogenic expansions of the GAA trinucleotide repeat in intron 1 of the FXN gene that lead to a decrease in frataxin protein levels. Rarely, affected individuals carry either a large intragenic deletion or whole-gene deletion of FXN on one allele and a full-penetrance expanded GAA repeat on the other allele.Case presentation We report here a patient that presented the typical clinical features of FRDA and genetic analysis of FXN intron 1 led to the assumption that the patient carried the common biallelic expansion. Subsequently, parental sample testing led to the identification of a novel intragenic deletion involving the 5'UTR upstream region and exons 1 and 2 of the FXN gene by MLPA.Conclusions With this case, we want to raise awareness about the potentially higher prevalence of intragenic deletions and underline the essential role of parental sample testing in providing accurate genetic counselling

Precipitated sdLDL: An easy method to estimate LDL particle size

Background: LDL-C lowering is the main measure in cardiovascular disease prevention but a residual risk of ischemic events still remains. Alterations of lipoproteins, specially, increase in small dense LDL (sdLDL) particles are related to this risk. Objective: To investigate the potential use of sdLDL cholesterol concentration (sdLDL-C) isolated by an easy precipitation method and to assess the impact of a set of clinical and biochemical variables determined by NMR on sdLDL concentration. Methods: sdLDL-C and NMR lipid profile were performed in 85 men samples. Association among them was evaluated using Pearson coefficients (rxy ). A multivariate regression was performed to identify the influence of NMR variables on sdLDL-C. Results: A strong association between sdLDL-C and LDLLDL-P (rxy = 0.687) and with LDL-Z (rxy = -0.603) was found. The multivariate regression explained a 56.8% in sdLDL-C variation (P = 8.77.10-12). BMI, ApoB, triglycerides, FFA, and LDL-Z showed a significant contribution. The most important ones were ApoB and LDL-Z; a 1nm increase (LDL-Z) leads to decrease 126 nmol/L in sdLDL-C. Conclusion: The association between sdLDL-C, LDL-Z, and LDL-P is clear. From a large number of variables, especially LDL-Z and apoB influence on sdLDL-C. Results show that the smaller the LDL size, the higher their cholesterol concentration. Therefore, sdLDL-C determination by using this easy method would be useful to risk stratification and to uncover cardiovascular residual risk

Reference values assessment in a Mediterranean population for small dense low-density lipoprotein concentration isolated by an optimized precipitation method

Background: High serum concentrations of small dense low-density lipoprotein cholesterol (sd-LDL-c) particles are associated with risk of cardiovascular disease (CVD). Their clinical application has been hindered as a consequence of the laborious current method used for their quantification. Objective: Optimize a simple and fast precipitation method to isolate sd-LDL particles and establish a reference interval in a Mediterranean population. Materials and methods: Forty-five serum samples were collected, and sd-LDL particles were isolated using a modified heparin-Mg2+ precipitation method. sd-LDL-c concentration was calculated by subtracting high-density lipoprotein cholesterol (HDL-c) from the total cholesterol measured in the supernatant. This method was compared with the reference method (ultracentrifugation). Reference values were estimated according to the Clinical and Laboratory Standards Institute and The International Federation of Clinical Chemistry and Laboratory Medicine recommendations. sd-LDL-c concentration was measured in serums from 79 subjects with no lipid metabolism abnormalities. Results: The Passing-Bablok regression equation is y = 1.52 (0.72 to 1.73) + 0.07x (−0.1 to 0.13), demonstrating no significant statistical differences between the modified precipitation method and the ultracentrifugation reference method. Similarly, no differences were detected when considering only sd-LDL-c from dyslipidemic patients, since the modifications added to the precipitation method facilitated the proper sedimentation of triglycerides and other lipoproteins. The reference interval for sd-LDL-c concentration estimated in a Mediterranean population was 0.04-0.47 mmol/L. Conclusion: An optimization of the heparin-Mg2+ precipitation method for sd-LDL particle isolation was performed, and reference intervals were established in a Spanish Mediterranean population. Measured values were equivalent to those obtained with the reference method, assuring its clinical application when tested in both normolipidemic and dyslipidemic subjects

Data on genotypic distribution and linkage disequilibrium of several ANRIL polymorphisms in hemodialysis patients

A long non-coding RNA called ANRIL located on chromosome 9p21.3 has been identified as a novel genetic factor associated with cardiovascular disease. Investigation of several single nucleotide polymorphisms (SNPs) of Noncoding Antisense RNA in the INK4 Locus (ANRIL) gene are of particular interest. This article reports data related to the research article entitled: "Association of ANRIL gene polymorphisms with major adverse cardiovascular events in hemodialysis patients" (Arbiol-Roca et al. [1]). Data presented show the genotypic distribution of four selected ANRIL SNPs: rs10757278, rs4977574, rs10757274 and rs6475606 in a cohort constituted by 284 hemodialysis patients. This article analyzes the Hardy-Weinberg disequilibrium of each studied SNP, and the linkage disequilibrium between them

How the analysis of archival data could provide helpful information about TID degradation. Case study: Bipolar transistors

A critical step of radiation hardness assurance (RHA) for space systems is given by the parts selection in accordance with the observed (or estimated) radiation effects. Although radiation testing is the most decisive way of studying the radiation degradation of electronic components, the increasing use of commercial off-the-shelf (COTS) devices and the challenges posed by NewSpace are pushing the need of finding new approaches to assess the risk associated with radiation environments. This work tries to evaluate if valuable information might be extracted from archival data to carry out this assessment despite the well-known and dramatic lot-to-lot, or even part-to-part, variability for some technologies and the impact of the different test conditions, such as the bias conditions and the dose rate in enhanced low dose rate sensitivity (ELDRS). These factors are briefly analyzed for some examples. A new radiation database is briefly introduced, and some statistical approaches are cited, apart from the analysis herein followed. To finish, a first analysis on three families of bipolar transistors is presented together with the independent results from three external reports, with a good agreement between the experimental results and the expected ones.10.13039/501100002878-Junta de Andalucia and Fondo Europeo de Desarrollo Regional (FEDER) Funds through the Singular Project Predicción del Comportamiento Eléctrico de Dispositivos Electrónicos bajo Radiación (PRECEDER) (Grant Number: CEI-5-RNM138). 10.13039/501100004837-Spanish Ministry of Science and Innovation under Project (Grant Number: PID2019-108377RB-C32)Peer reviewe

Identificación de genes implicados en la adquisición del fenotipo metastásico en modelos experimentales de cáncer de mama

Consultable des del TDXTítol obtingut de la portada digitalitzadaEl cáncer de mama es una de las enfermedades más frecuentes entre las mujeres de los países desarrollados. La metástasis de los tumores es la principal causa de la muerte de un paciente con cáncer. Una de las maneras de abordar el estudio molecular y celular del proceso de desarrollo de las metástasis es la identificación de genes asociados al fenotipo metastásico. El método RAP-PCR (RNA arbitrarily primed PCR) consiste en dos reacciones consecutivas: una transcripción inversa del mRNA utilizando un cebador aleatorio y una reacción en cadena de la polimerasa (PCR) en que se emplea el mismo cebador más otro cualquiera. Con este método se pueden comparar simultáneamente fragmentos de DNA obtenidos aleatoriamente a partir del RNA de muchas variantes celulares con diferente capacidad metastásica. Si estos fragmentos, usados como sondas en análisis Northern, hibridan con RNA y se observan en cantidades relativas muy diferentes entre las distintas variantes celulares, se asume que los genes cuya expresión representan se expresan de manera muy diferente y pueden ser, por tanto, candidatos a influir en la aparición del fenotipo metastásico. En el presente trabajo, se ha empleado este método para el estudio de dos modelos experimentales de carcinoma de mama: uno de ratón (MXT) y otro humano (MDA MB-435). En ambos modelos existen variantes celulares con características invasivas y metastásicas bien diferenciadas. Los resultados obtenidos muestran que el método RAP-PCR permite detectar RNAs anónimos correspondientes a genes que se expresan diferencialmente en variantes de distinta capacidad metastásica, pero de acervo genético común. A partir de algunos de los fragmentos observados con expresión diferencial en los modelos tumorales MXT y MDA MB-435 (más de 50), se han identificado diversos genes. Los de mayor expresión en las variantes metastásicas fueron los correspondientes a enolasa, ESDN (endothelial and smooth muscle cell-derived neuropilin-like protein), S-adenosilhomocisteína hidrolasa, CHL1 (close homolog to L1CAM), Slmap (sarcolemmal-associated protein), eEF-1α (factor de elongación 1α), Nono (non-POU domain-containing octamer-binding protein), moesina, eplin (epithelial protein lost in neoplasm), ADAMTS1, y a la proteína hipotética FLJ10830. Los de menor expresión en las variantes metastásicas fueron los correspondientes a la cadena β de HLA-DP, la enzima E2 (variante 1) (ubiquitin-conjugating enzyme E2 variant 1), y el gen denominado 7MXT en el presente trabajo. La función de estos genes puede adscribirse a alguna de estas categorías: metabolismo celular, regulación de la expresión génica, angiogénesis / diferenciación / invasión, proliferación celular; además, hay genes aún no identificados o de función todavía no conocida. El gen denominado 7MXT se encuentra en el cromosoma 11 del ratón, pero todavía no ha sido identificado. Su expresión está claramente disminuida en las variantes más metastásicas, pero se expresa en grado diverso en todos los órganos embrionarios de ratón en que se ha probado; la expresión más abudante corresponde al bazo y al corazón. En los ratones adultos sólo se ha detectado su expresión en el bazo y, sobre todo, en el ovario. La disminución de la expresión de ADAMTS1 hace aumentar la capacidad migratoria de las células tumorales B2 a través del colágeno IV (si bien no modifica la capacidad de adhesión de estas células a los distintos componentes de la matriz extracelular); además, aumenta el tiempo de latencia de crecimiento de los tumores primarios, pero no modifica la capacidad metastásica. Existe una conexión entre la aparición del fenotipo metastásico y la ausencia de expresión del gen de la cadena β de HLA, una proteína del MHC-II que no se había asociado previamente con la metástasis. Además, la expresión de CIITA (transactivador del MHC-II), determina la expresión de MHC-II en las variantes metastásicas.Breast cancer is one of the most frequent malignancies in women from developed countries. Tumor metastasis is the main cause of death in cancer patients. A useful approach to study the molecular and cellular process of metastasis development is to identify genes related to metastatic phenotype. RAP-PCR (RNA arbitrarily primed PCR) method consists of two consecutive reactions: a reverse transcription of mRNA usin an arbitrary primer and a polymerase chain reaction (PCR) using the same primer and any other one. With this method random DNA fragments obtained from RNA from many cell variants of different metastasic ability can be simultaneusly compared. When these fragments -used as probes in Northern blots- hybridize to real RNA and they are observed in different relative ammounts between the cell variants, the genes whose expression they represent are assumed to be differentially expressed and can be considered as candidates to be involved in metastasic phenotype development. In the present work, this method was employed to study two experimental models of breast carcinoma: a murine one (MXT) and a human one (MDA MB-435). Both models include cell variants with different invasive and metastatic abilities. The results show that RAP-PCR method let us detect anonyme RNAs corresponding to genes differentially expressed in cell variants of different metastatic ability, but sharing a common genetic background. Several genes were identified that showed differnetial expression in tumor models MXT and MDA MB-435. Those with a higher expression in metastatic cell variants were: enolase, ESDN (endothelial and smooth muscle cell-derived neuropilin-like protein), S-adenosilhomocystein hydrolase, CHL1 (close homolog to L1CAM), Slmap (sarcolemmal-associated protein), eEF-1α (elongation factor 1α), Nono (non-POU domain-containing octamer-binding protein), moesin, eplin (epithelial protein lost in neoplasm), ADAMTS1, and hypothetic protein FLJ10830. Those with a lower expression in metastatic cell variants were: HLA-DP β−chain, E2 enzyme (variant 1) (ubiquitin-conjugating enzyme E2 variant 1), and the gene here called 7MXT. The function of these genes can be abscribed to one of the following cathegories: cellular metabolism, gene expression regulation, angiogenesis/differentiatio/invasion, cellular proliferation. Furthermore, several genes had not yet been identified or had an unknown function. The so-called 7MXT gene is located in chromose 11 in mice, but has not yet been identified. Its expression is clearly diminished in highly metastatic cell variants, but all of the tested embrionary organs express it in variable degree, and above all spleen and heart. Expression in adult mice has only been demonstrated in spleen and especially ovary. The decrease in ADAMTS-1 expression increases the migratory ability of highly metastatic tumor cells MXT-B2 through collagen IV (though no difference in adhesion ability to several extracellular matrix elements is observed); furthermore, ADAMTS-1 decrease increases latency in the development of primary tumors, without modifying the metastatic ability. There is a relation between the metastatic phenotype and the loss of expression of HLA-DP β-chain gene -an MHC-II protein not previously associated to metastasis. Moreover, the expression of CIITA (an MHC-II transactivator) determines MHC-II expression in metastatic cell variants

Identificación de genes implicados en la adquisición del fenotipo metastásico en modelos experimentales de cáncer de mama

El cáncer de mama es una de las enfermedades más frecuentes entre las mujeres de los países desarrollados. La metástasis de los tumores es la principal causa de la muerte de un paciente con cáncer. Una de las maneras de abordar el estudio molecular y celular del proceso de desarrollo de las metástasis es la identificación de genes asociados al fenotipo metastásico.El método RAP-PCR (RNA arbitrarily primed PCR) consiste en dos reacciones consecutivas: una transcripción inversa del mRNA utilizando un cebador aleatorio y una reacción en cadena de la polimerasa (PCR) en que se emplea el mismo cebador más otro cualquiera. Con este método se pueden comparar simultáneamente fragmentos de DNA obtenidos aleatoriamente a partir del RNA de muchas variantes celulares con diferente capacidad metastásica. Si estos fragmentos, usados como sondas en análisis Northern, hibridan con RNA y se observan en cantidades relativas muy diferentes entre las distintas variantes celulares, se asume que los genes cuya expresión representan se expresan de manera muy diferente y pueden ser, por tanto, candidatos a influir en la aparición del fenotipo metastásico.En el presente trabajo, se ha empleado este método para el estudio de dos modelos experimentales de carcinoma de mama: uno de ratón (MXT) y otro humano (MDA MB-435). En ambos modelos existen variantes celulares con características invasivas y metastásicas bien diferenciadas.Los resultados obtenidos muestran que el método RAP-PCR permite detectar RNAs anónimos correspondientes a genes que se expresan diferencialmente en variantes de distinta capacidad metastásica, pero de acervo genético común.A partir de algunos de los fragmentos observados con expresión diferencial en los modelos tumorales MXT y MDA MB-435 (más de 50), se han identificado diversos genes. Los de mayor expresión en las variantes metastásicas fueron los correspondientes a enolasa, ESDN (endothelial and smooth muscle cell-derived neuropilin-like protein), S-adenosilhomocisteína hidrolasa, CHL1 (close homolog to L1CAM), Slmap (sarcolemmal-associated protein), eEF-1&#945; &#61472;(factor de elongación 1&#945;), Nono (non-POU domain-containing octamer-binding protein), moesina, eplin (epithelial protein lost in neoplasm), ADAMTS1, y a la proteína hipotética FLJ10830.Los de menor expresión en las variantes metastásicas fueron los correspondientes a la cadena &#946; de HLA-DP, la enzima E2 (variante 1) (ubiquitin-conjugating enzyme E2 variant 1), y el gen denominado 7MXT en el presente trabajo.La función de estos genes puede adscribirse a alguna de estas categorías: metabolismo celular, regulación de la expresión génica, angiogénesis / diferenciación / invasión, proliferación celular; además, hay genes aún no identificados o de función todavía no conocida.El gen denominado 7MXT se encuentra en el cromosoma 11 del ratón, pero todavía no ha sido identificado. Su expresión está claramente disminuida en las variantes más metastásicas, pero se expresa en grado diverso en todos los órganos embrionarios de ratón en que se ha probado; la expresión más abudante corresponde al bazo y al corazón. En los ratones adultos sólo se ha detectado su expresión en el bazo y, sobre todo, en el ovario. La disminución de la expresión de ADAMTS1 hace aumentar la capacidad migratoria de las células tumorales B2 a través del colágeno IV (si bien no modifica la capacidad de adhesión de estas células a los distintos componentes de la matriz extracelular); además, aumenta el tiempo de latencia de crecimiento de los tumores primarios, pero no modifica la capacidad metastásica.Existe una conexión entre la aparición del fenotipo metastásico y la ausencia de expresión del gen de la cadena &#946; de HLA, una proteína del MHC-II que no se había asociado previamente con la metástasis. Además, la expresión de CIITA (transactivador del MHC-II), determina la expresión de MHC-II en las variantes metastásicas.Breast cancer is one of the most frequent malignancies in women from developed countries. Tumor metastasis is the main cause of death in cancer patients. A useful approach to study the molecular and cellular process of metastasis development is to identify genes related to metastatic phenotype.RAP-PCR (RNA arbitrarily primed PCR) method consists of two consecutive reactions: a reverse transcription of mRNA usin an arbitrary primer and a polymerase chain reaction (PCR) using the same primer and any other one. With this method random DNA fragments obtained from RNA from many cell variants of different metastasic ability can be simultaneusly compared. When these fragments -used as probes in Northern blots- hybridize to real RNA and they are observed in different relative ammounts between the cell variants, the genes whose expression they represent are assumed to be differentially expressed and can be considered as candidates to be involved in metastasic phenotype development.In the present work, this method was employed to study two experimental models of breast carcinoma: a murine one (MXT) and a human one (MDA MB-435). Both models include cell variants with different invasive and metastatic abilities.The results show that RAP-PCR method let us detect anonyme RNAs corresponding to genes differentially expressed in cell variants of different metastatic ability, but sharing a common genetic background.Several genes were identified that showed differnetial expression in tumor models MXT and MDA MB-435.Those with a higher expression in metastatic cell variants were: enolase, ESDN (endothelial and smooth muscle cell-derived neuropilin-like protein), S-adenosilhomocystein hydrolase, CHL1 (close homolog to L1CAM), Slmap (sarcolemmal-associated protein), eEF-1&#945; &#61472;(elongation factor 1&#945;), Nono (non-POU domain-containing octamer-binding protein), moesin, eplin (epithelial protein lost in neoplasm), ADAMTS1, and hypothetic protein FLJ10830.Those with a lower expression in metastatic cell variants were: HLA-DP &#946;&#8722;chain, E2 enzyme (variant 1) (ubiquitin-conjugating enzyme E2 variant 1), and the gene here called 7MXT.The function of these genes can be abscribed to one of the following cathegories: cellular metabolism, gene expression regulation, angiogenesis/differentiatio/invasion, cellular proliferation. Furthermore, several genes had not yet been identified or had an unknown function.The so-called 7MXT gene is located in chromose 11 in mice, but has not yet been identified. Its expression is clearly diminished in highly metastatic cell variants, but all of the tested embrionary organs express it in variable degree, and above all spleen and heart. Expression in adult mice has only been demonstrated in spleen and especially ovary.The decrease in ADAMTS-1 expression increases the migratory ability of highly metastatic tumor cells MXT-B2 through collagen IV (though no difference in adhesion ability to several extracellular matrix elements is observed); furthermore, ADAMTS-1 decrease increases latency in the development of primary tumors, without modifying the metastatic ability.There is a relation between the metastatic phenotype and the loss of expression of HLA-DP &#946;-chain gene -an MHC-II protein not previously associated to metastasis. Moreover, the expression of CIITA (an MHC-II transactivator) determines MHC-II expression in metastatic cell variants

Measurement of sirolimus concentrations in human blood using an automated electrochemiluminescence immunoassay (ECLIA): a multicenter evaluation.

Background: Therapeutic drug monitoring (TDM) of sirolimus is essential in transplant recipients. We evaluated the performance of a new electrochemiluminescence immunoassay (ECLIA) for measuring sirolimus concentrations in whole blood at five European laboratories. Methods: Study assessments included repeatability, intermediate precision and functional sensitivity (concentration at coefficient of variation [CV] of 20%) experiments. Method comparisons with liquid chromatography-tandem mass spectrometry (LC-MS/MS; reference method) and two immunoassays (chemiluminescent microparticle immunoassay [CMIA] and antibody-conjugated magnetic immunoassay [ACMIA]) were performed using native samples from patients with kidney transplants. Results: Imprecision testing CVs were ≤6.4% and ≤10.7% across the sirolimus concentration range for both repeatability and intermediate precision, respectively. The ECLIA showed excellent functional sensitivity: the CV did not reach 20%; the CV at the assay's limit of quantitation (1.5 μg/L) was 7.0%. Agreement between the ECLIA and LC-MS/MS using native kidney samples was close, with weighted Deming regression analysis yielding a slope of 1.05, an intercept of 0.154 μg/L and a Pearson's correlation coefficient (r) of 0.94, while Bland-Altman analysis showed a combined mean bias of 0.41 μg/L (±2 standard deviation [SD], -1.96 to 2.68). The ECLIA also showed good correlation with the two other immunoassays: the CMIA (slope=0.91, intercept=0.112 μg/L and r=0.89) and the ACMIA (slope=0.99, intercept=0.319 μg/L and r=0.97). Conclusions: The ECLIA showed good precision, functional sensitivity and agreement with other methods of sirolimus measurement used in clinical practice, suggesting that the assay is suitable for TDM in transplant recipients and provides an alternative to LC-MS/M