RESUMEN
BACKGROUND: Health education interventions are considered critical for the prevention and management of conditions of public health concern. Although the burden of these conditions is often greatest in socio-economically disadvantaged populations, the effectiveness of interventions that target these groups is unknown. We aimed to identify and synthesize evidence of the effectiveness of health-related educational interventions in adult disadvantaged populations. METHODS: We pre-registered the study on Open Science Framework https://osf.io/ek5yg/ . We searched Medline, Embase, Emcare, and the Cochrane Register from inception to 5/04/2022 to identify studies evaluating the effectiveness of health-related educational interventions delivered to adults in socio-economically disadvantaged populations. Our primary outcome was health related behaviour and our secondary outcome was a relevant biomarker. Two reviewers screened studies, extracted data and evaluated risk of bias. Our synthesis strategy involved random-effects meta-analyses and vote-counting. RESULTS: We identified 8618 unique records, 96 met our criteria for inclusion - involving more than 57,000 participants from 22 countries. All studies had high or unclear risk of bias. For our primary outcome of behaviour, meta-analyses found a standardised mean effect of education on physical activity of 0.05 (95% confidence interval (CI) = -0.09-0.19), (5 studies, n = 1330) and on cancer screening of 0.29 (95% CI = 0.05-0.52), (5 studies, n = 2388). Considerable statistical heterogeneity was present. Sixty-seven of 81 studies with behavioural outcomes had point estimates favouring the intervention (83% (95% CI = 73%-90%), p < 0.001); 21 of 28 studies with biomarker outcomes showed benefit (75% (95%CI = 56%-88%), p = 0.002). When effectiveness was determined based on conclusions in the included studies, 47% of interventions were effective on behavioural outcomes, and 27% on biomarkers. CONCLUSIONS: Evidence does not demonstrate consistent, positive impacts of educational interventions on health behaviours or biomarkers in socio-economically disadvantaged populations. Continued investment in targeted approaches, coinciding with development of greater understanding of factors determining successful implementation and evaluation, are important to reduce inequalities in health.
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Educación en Salud , Poblaciones Vulnerables , Adulto , Humanos , Promoción de la Salud , Sesgo , Conductas Relacionadas con la SaludRESUMEN
Genome-wide association studies (GWAS) have identified a region upstream the BIN1 gene as the most important genetic susceptibility locus in Alzheimer's disease (AD) after APOE. We report that BIN1 transcript levels were increased in AD brains and identified a novel 3 bp insertion allele â¼28 kb upstream of BIN1, which increased (i) transcriptional activity in vitro, (ii) BIN1 expression levels in human brain and (iii) AD risk in three independent case-control cohorts (Meta-analysed Odds ratio of 1.20 (1.14-1.26) (P=3.8 × 10(-11))). Interestingly, decreased expression of the Drosophila BIN1 ortholog Amph suppressed Tau-mediated neurotoxicity in three different assays. Accordingly, Tau and BIN1 colocalized and interacted in human neuroblastoma cells and in mouse brain. Finally, the 3 bp insertion was associated with Tau but not Amyloid loads in AD brains. We propose that BIN1 mediates AD risk by modulating Tau pathology.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/genética , Enfermedad de Alzheimer/genética , Enfermedad de Alzheimer/patología , Predisposición Genética a la Enfermedad/genética , Proteínas Nucleares/genética , Proteínas Supresoras de Tumor/genética , Proteínas tau/metabolismo , Proteínas Adaptadoras Transductoras de Señales/biosíntesis , Enfermedad de Alzheimer/metabolismo , Animales , Encéfalo/metabolismo , Encéfalo/patología , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , Estudios de Casos y Controles , Células Cultivadas , Proteínas de Drosophila/deficiencia , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Endofenotipos , Expresión Génica/genética , Humanos , Ratones , Degeneración Nerviosa/genética , Degeneración Nerviosa/patología , Proteínas Nucleares/biosíntesis , Placa Amiloide/patología , Polimorfismo de Nucleótido Simple/genética , Sinaptosomas/patología , Factores de Transcripción/deficiencia , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Proteínas Supresoras de Tumor/biosíntesis , Proteínas tau/antagonistas & inhibidoresRESUMEN
Chromosomal translocations involving the immunoglobulin heavy chain (IGH) locus define common subgroups of B-cell lymphoma but are rare in B-cell precursor acute lymphoblastic leukemia (BCP-ALL). Recent fluorescent in situ hybridization and molecular cloning studies have identified several novel IGH translocations involving genes that play important roles in normal hemopoiesis, including the cytokine receptor genes CRLF2 and EPOR, all members of the CCAAT enhancer-binding protein gene family, as well as genes not normally expressed in hemopoietic cells including inhibitor of DNA binding 4. IGH translocation results in deregulated target gene expression because of juxtaposition with IGH transcriptional enhancers. However, many genes targeted by IGH translocations are also more commonly deregulated in BCP-ALL as a consequence of other genetic or epigenetic mechanisms. For example, interstitial genomic deletions also result in deregulated CRLF2 expression, whereas EPOR expression is deregulated as a consequence of the ETV6-RUNX1 fusion. The possible clinical importance of many of the various IGH translocations in BCP-ALL remains to be determined from prospective studies, but CRLF2 expression is associated with a poor prognosis. Despite their rarity, IGH chromosomal translocations in BCP-ALL therefore define not only new mechanisms of B-cell transformation but also clinically important subgroups of disease and suggest new targeted therapeutic approaches.
Asunto(s)
Linfocitos B/metabolismo , Transformación Celular Neoplásica/metabolismo , Cadenas Pesadas de Inmunoglobulina/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras B/metabolismo , Sitios de Carácter Cuantitativo , Translocación Genética , Enfermedad Aguda , Animales , Transformación Celular Neoplásica/genética , Subunidad alfa 2 del Factor de Unión al Sitio Principal/biosíntesis , Subunidad alfa 2 del Factor de Unión al Sitio Principal/genética , Regulación Leucémica de la Expresión Génica/genética , Hematopoyesis/genética , Humanos , Cadenas Pesadas de Inmunoglobulina/genética , Proteínas Inhibidoras de la Diferenciación/biosíntesis , Proteínas Inhibidoras de la Diferenciación/genética , Proteínas de Fusión Oncogénica/biosíntesis , Proteínas de Fusión Oncogénica/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras B/diagnóstico , Leucemia-Linfoma Linfoblástico de Células Precursoras B/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras B/terapia , Pronóstico , Receptores de Citocinas/biosíntesis , Receptores de Citocinas/genética , Receptores de Eritropoyetina/biosíntesis , Receptores de Eritropoyetina/genéticaRESUMEN
Molecular cloning of immunoglobulin heavy chain (IGH) translocation breakpoints identifies genes of biological importance in the development of normal and malignant B cells. Long-distance inverse PCR (LDI-PCR) was first applied to amplification of IGH gene translocations targeted to the joining (IGHJ) regions. We report here successful amplification of the breakpoint of IGH translocations targeted to switch (IGHS) regions by LDI-PCR. To detect IGHS translocations, Southern blot assays using 5' and 3' switch probes were performed. Illegitimate Smu rearrangements were amplified from the 5' end (5'Smu LDI-PCR) from the alternative derivative chromosome, and those of Sgamma or Salpha were amplified from the 3' end (3'Sgamma or 3'alpha LDI-PCR) from the derivative chromosome 14. Using a combination of these methods, we have succeeded in amplifying IGHS translocation breakpoints involving FGFR3/MMSET on 4p16, BCL6 on 3q27, MYC on 8q24, IRTA1 on 1q21 and PAX5 on 9p13 as well as BCL11A on 2p13 and CCND3 on 6p21. The combination of LDI-PCR for IGHJ and IGHS allows rapid molecular cloning of almost all IGH gene translocation breakpoints.
Asunto(s)
Rotura Cromosómica/genética , Reordenamiento Génico , Genes de Cambio/genética , Cadenas Pesadas de Inmunoglobulina/genética , Región de Cambio de la Inmunoglobulina/genética , Linfoma/genética , Translocación Genética , Secuencia de Bases , Southern Blotting , Proteínas Portadoras/genética , Cromosomas Humanos Par 14 , Clonación Molecular , Proteínas de Unión al ADN/genética , Humanos , Cadenas mu de Inmunoglobulina/genética , Datos de Secuencia Molecular , Proteínas Nucleares/genética , Factor de Transcripción PAX5 , Reacción en Cadena de la Polimerasa , Proteínas Tirosina Quinasas/genética , Proteínas Proto-Oncogénicas/genética , Proteínas Proto-Oncogénicas c-bcl-6 , Proteínas Proto-Oncogénicas c-myc/genética , Receptor Tipo 3 de Factor de Crecimiento de Fibroblastos , Receptores de Factores de Crecimiento de Fibroblastos/genética , Proteínas Represoras , Factores de Transcripción/genética , Células Tumorales CultivadasRESUMEN
B-cell-specific plasma-membrane proteins are potential targets for either small molecule or antibody-based therapies. We have sought to annotate proteins expressed at the cell surface membrane in patients with chronic lymphocytic leukemia (CLL) using plasma-membrane-based proteomic analysis to identify previously uncharacterized and potentially B-cell-specific proteins. Proteins from plasma-membrane fractions were separated on one-dimensional gels and trypsinized fractions subjected to high-throughput MALDI-TOF mass spectrometry. Using this method, many known B-cell surface antigens were detected, but also known proteins not previously described in this disease or in this cellular compartment, including cell surface receptors, membrane-associated enzymes and secreted proteins, and completely unknown proteins. To validate the method, we show that BLK, a B-cell-specific kinase, is located in the CLL-plasma-membrane fraction. We also describe two novel proteins (MIG2B and B-cell novel protein #1, BCNP1), which are expressed preferentially in B cells. MIG2B is in a highly conserved and defined gene family containing two plasma-membrane-binding ezrin/radixin/moesin domains and a pleckstrin homology domain; the Caenorhabditis elegans homolog (UNC-112) is a membrane-associated protein that colocalizes with integrin at cell-matrix adhesion complexes. BCNP1 is a completely unknown protein with three predicted transmembrane domains, with three alternatively spliced final exons. Proteomic analysis may thus define new potential therapeutic targets.
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Linfocitos B/química , Leucemia Linfocítica Crónica de Células B/patología , Proteínas de la Membrana/aislamiento & purificación , Proteínas de Neoplasias/aislamiento & purificación , Proteómica , Proteínas Reguladoras de la Apoptosis , Linfocitos B/patología , Secuencia de Bases , Western Blotting , Humanos , Proteínas de la Membrana/química , Proteínas de la Membrana/genética , Datos de Secuencia Molecular , Proteínas de Neoplasias/química , Proteínas de Neoplasias/genética , Sistemas de Lectura Abierta , Isoformas de Proteínas , Estructura Terciaria de Proteína , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
Many subtypes of B-cell malignancy are characterized by chromosomal translocations that target the immunoglobulin loci. Molecular cloning of such translocations continues to allow the identification of genes whose deregulated expression plays a pivotal role in the pathogenesis of B-cell malignancy. The clustering of breakpoints within the immunoglobulin loci has allowed the development of rapid and robust polymerase chain reaction methods for cloning. We discuss in this chapter the use of long-distance inverse polymerase chain reaction methods to clone immunoglobulin chromosomal translocation breakpoints from clinical material. These methods have been successfully applied to several other types of chromosomal translocation including those involving other genes such as BCL6, ETV6, and MYC.
Asunto(s)
Rotura Cromosómica , Reordenamiento Génico , Genes de Inmunoglobulinas , Reacción en Cadena de la Polimerasa/métodos , Translocación Genética , Cromosomas Humanos/genética , Clonación Molecular , Proteínas de Unión al ADN/genética , Genes myc/fisiología , Humanos , Linfoma no Hodgkin/genética , Proteínas Nucleares/genética , Proteínas Proto-Oncogénicas/genética , Proteínas Proto-Oncogénicas c-bcl-6 , Proteínas Proto-Oncogénicas c-ets , Proteínas Represoras/genética , Factores de Transcripción/genética , Proteína ETS de Variante de Translocación 6RESUMEN
Angiotensin converting enzyme (ACE) and the Alzheimer's amyloid precursor protein are cleaved from the membrane by zinc metalloproteinases termed ACE secretase and alpha-secretase, respectively. Tumour necrosis factor-alpha (TNF-alpha) convertase (ADAM 17) is a recently identified member of the adamalysin family of mammalian zinc metalloproteinases that is involved in the production of TNF-alpha and possibly in the cleavage of other membrane proteins. Using two different cell-free assays we were unable to detect significant cleavage and secretion of ACE by TNF-alpha convertase. In addition, there was a different effect of three hydroxamic acid-based inhibitors (batimastat, compound 1 and compound 4) towards TNF-alpha convertase as compared to ACE secretase and alpha-secretase. Thus TNF-alpha convertase would appear to be distinct from, but possibly related to, the secretases that cleave ACE and the amyloid precursor protein.
Asunto(s)
Precursor de Proteína beta-Amiloide/metabolismo , Endopeptidasas/metabolismo , Metaloendopeptidasas/metabolismo , Peptidil-Dipeptidasa A/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Proteínas ADAM , Proteína ADAM17 , Secretasas de la Proteína Precursora del Amiloide , Animales , Sistema Libre de Células , Metaloendopeptidasas/antagonistas & inhibidores , PorcinosRESUMEN
Matrix metalloproteinases (MMPs) and the specific tissue inhibitors of metalloproteinases (TIMPs) are involved in tissue turnover in normal and pathological processes including wound healing. Marimastat, a potent inhibitor of MMPs, was used to investigate the role of MMPs in an in vitro wound contraction model, the dermal equivalent, in which fibroblasts are grown in a collagen matrix. Marimastat inhibited fibroblast-mediated lattice contraction and this inhibition was reversible upon removal of the inhibitor, indicating that MMPs play an important role in fibroblast-mediated collagen lattice contraction, modelling what may happen when granulation tissue contracts in a healing wound.
Asunto(s)
Colágeno/fisiología , Colagenasas/metabolismo , Ácidos Hidroxámicos/farmacología , Metaloendopeptidasas/antagonistas & inhibidores , Piel/metabolismo , Adulto , Células Cultivadas , Colágeno/efectos de los fármacos , Fibroblastos/metabolismo , Humanos , Metaloproteinasa 1 de la Matriz , Piel/citología , Piel/efectos de los fármacosRESUMEN
The synthesis of a series of thiol-containing, modified dipeptide inhibitors (8) of human collagenase, which incorporate various carboxylic acid derivatives at the presumed P1 position, beta to the thiol group, is described. The compounds were evaluated, in vitro, for their ability to inhibit the degradation of rat skin type 1 collagen by purified human lung fibroblast collagenase, and structure-activity relationship studies are described. Optimum potency (IC50 values in the nanomolar range) was achieved by incorporating methyl (compounds 43a, 56a, and 57ab) or benzyl esters (44a) at the P1 position. Small amides were also accommodated (e.g. primary amide 47a), but in general, increasing the size of the P1 amide substituent lowered potency. PheNHMe, TrpNHMe, and Tyr(Me)NHMe substituents were found to be approximately equipotent P2'-residues. The results of testing all four diastereoisomers 56a-d of the compound with (S)-TrpNHMe at the P2' position indicated that the S,S,S diastereoisomer 56a possessed highest potency (IC50 2.5 nM) and that the second most potent diastereoisomer was 56d (IC50 12 nM) with the R,R,S configuration. It appeared that the orientation of the P1' and the thiol-bearing centers to each other is a more critical influence on potency than any absolute stereochemical requirements. It is suggested that the high potency of the beta-mercapto carboxylic acid derivatives may be a consequence of bidentate coordination of the thiol and carbonyl groups to the active-site zinc ion in the collagenase enzyme.
Asunto(s)
Ácidos Carboxílicos/síntesis química , Inhibidores de la Metaloproteinasa de la Matriz , Compuestos de Sulfhidrilo/síntesis química , Animales , Ácidos Carboxílicos/química , Ácidos Carboxílicos/farmacología , Colagenasas/metabolismo , Fibroblastos/efectos de los fármacos , Fibroblastos/enzimología , Humanos , Ratas , Estereoisomerismo , Relación Estructura-Actividad , Compuestos de Sulfhidrilo/química , Compuestos de Sulfhidrilo/farmacologíaRESUMEN
The synthesis of a series of N-phosphonalkyl dipeptides 6 is described. Syntheses were devised that allowed the preparation of single diastereoisomers and the assignment of stereochemistry. The compounds were evaluated in vitro for their ability to inhibit the degradation of radiolabeled collagen by purified human lung fibroblast collagenase. Several of the compounds were potent collagenase inhibitors and were at least 10-fold more potent than their corresponding N-carboxyalkyl analogues. Activity was lost when the phosphonic acid group P(O)(OH)2 was replaced by the phosphinic acid groups P(O)(H)(OH) and P(O)(Me)(OH). At the P1 position, (R)- or (S)-alkyl groups, especially ethyl and methyl (e.g., 12a,b, 52a,b, and 53a,b), or an (R)-phenethyl moiety (55a) conferred high potency (IC50 values in the range 0.23-0.47 microM). (S)-Stereochemistry was preferred for the P1' isobutyl side chain. Structure-activity relationships were also investigated at the P2' site, and interestingly, compounds with basic side chains, such as the guanidine 57a, were equipotent with more lipophilic compounds, such as 52a. As with other series of collagenase inhibitors, potency was enhanced by introducing bicyclic aromatic P2' substituents. The most potent phosphonic acid of the series was the bicyclic aromatic P2' tryptophan analogue 59a (IC50 0.05 microM).
Asunto(s)
Dipéptidos/síntesis química , Inhibidores de la Metaloproteinasa de la Matriz , Animales , Sitios de Unión , Ácidos Carboxílicos/química , Ácidos Carboxílicos/metabolismo , Línea Celular , Colágeno/metabolismo , Colagenasas/química , Colagenasas/metabolismo , Dipéptidos/farmacología , Fibroblastos/enzimología , Humanos , Conformación Molecular , Estructura Molecular , Organofosfonatos/química , Organofosfonatos/metabolismo , Peptidil-Dipeptidasa A/química , Ácidos Fosfínicos/química , Ácidos Fosfínicos/metabolismo , Ratas , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
Missense mutations in the presenilin-1 (PS-1) and presenilin-2 (PS-2) genes have been shown to be causes of autosomal dominant Alzheimer's disease (the AD3 and AD4 loci, respectively). Alternative splicing has previously been reported in the PS-1 gene. In this study, elucidation of intron/exon boundary sequences revealed that PS-2 is encoded by 10 coding exons. In addition, PS-2 cDNA cloning and RT-PCR using RNA from a variety of normal tissues revealed the presence of alternatively spliced products. These products included species with in frame omissions of exon 8 and simultaneous omissions of exons 3 and 4.
Asunto(s)
Empalme Alternativo/genética , Proteínas de la Membrana/genética , Enfermedad de Alzheimer/genética , Secuencia de Aminoácidos , Secuencia de Bases , Exones/genética , Intrones/genética , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Presenilina-2RESUMEN
The presenilin 1 gene has recently been identified as the locus on chromosome 14 which is responsible for a large proportion of early onset, autosomal dominantly inherited Alzheimer's disease (AD). We have elucidated the intron/exon structure of the gene and designed intronic primers to enable direct sequencing of the entire coding region (10 exons) of the presenilin gene in a large number of families. This strategy has enabled us to find a further two novel mutations in the gene. We discuss the distribution of mutations and the proportions of autosomal dominant AD with a mean age of onset below 60 years caused by mutations in this gene.
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Enfermedad de Alzheimer/genética , Proteínas de la Membrana/genética , Enfermedad de Alzheimer/metabolismo , Secuencia de Bases , Análisis por Conglomerados , Cartilla de ADN , Exones/fisiología , Ligamiento Genético , Genoma , Humanos , Irlanda , Proteínas de la Membrana/metabolismo , Datos de Secuencia Molecular , Mutación , Sistemas de Lectura Abierta , Presenilina-1 , Reino UnidoRESUMEN
The effects of lamotrigine on rat neuroma and behavioural paradigms were evaluated to determine a pre-clinical therapeutic index. Lamotrigine blocked neuroma-induced burst pattern firing at a free plasma concentration of 13.7+/-1.7 microM (n=5). Oral dosing of lamotrigine (50-200 mg/kg) had no significant effects on behaviour but measurements of plasma concentrations of free drug showed non-linear oral absorption and lower than predicted drug levels (5-27 microM). Given intravenously (10-100 mg/kg), lamotrigine did affect behaviour at a free plasma concentration of 42.0 microM (n=2). By comparing free plasma concentrations, a therapeutic index of 3 was calculated, which is lower than published data based on comparing oral doses. We propose that a therapeutic index should only be derived with reference to plasma drug concentrations to prevent non-linear or incomplete drug absorption from confounding accurate estimation.
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Potenciales de Acción/efectos de los fármacos , Anticonvulsivantes/farmacología , Conducta Animal/efectos de los fármacos , Neuroma/fisiopatología , Triazinas/farmacología , Potenciales de Acción/fisiología , Animales , Anticonvulsivantes/sangre , Anticonvulsivantes/uso terapéutico , Conducta Animal/fisiología , Evaluación Preclínica de Medicamentos , Lamotrigina , Masculino , Destreza Motora/efectos de los fármacos , Destreza Motora/fisiología , Neuroma/tratamiento farmacológico , Ratas , Ratas Sprague-Dawley , Nervio Ciático/efectos de los fármacos , Nervio Ciático/lesiones , Nervio Ciático/fisiología , Triazinas/sangre , Triazinas/uso terapéuticoRESUMEN
A straightforward in vivo model of collagen degradation is described that can be used to measure the effects of different classes of proteinase inhibitors. Air pouches, formed subcutaneously in the dorsal thoracic region of rats, were inflamed 6 to 8 days later by injecting lambda-type carrageenan. 14C-Collagen was injected into the air pouches either 1 day before or 1 day after lambda-carrageenan-induced inflammation: in the latter case, the inflammatory exudate fluid was drained from the air pouches immediately prior to administering 14C-collagen. Ninety percent of the 14C-collagen was degraded and cleared within 3 days from pre-inflamed air pouches, but degradation was much slower from the post-inflamed or non-inflamed air pouches. Proteinase inhibitors injected simultaneously with the 14C-collagen, and again 6 hr later, reduced the extent of 14C-collagen degradation from air pouches measured after 24 hr. Forty-two percent of the degradation of 14C-collagen could be inhibited by a mixture of enzyme inhibitors (leupeptin, alpha 1-anti-proteinase, aprotinin, and pepstatin) injected together with 1,10 phenanthroline, the zinc metalloenzyme inhibitor. The 1,10 phenanthroline alone caused a 33% inhibition of 14C-collagen degradation, and the inhibitor mixture given alone inhibited 14C-collagen loss by 25%. Approximately 60% of the degradation of 14C-collagen in this model was mediated by mechanisms resistant to this combination of proteinase inhibitors, which may indicate the significant involvement of non-enzymic modalities, or degradation in intracellular compartments inaccessible to extracellular agents.
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Colágeno/metabolismo , Inhibidores de Proteasas/farmacología , Piel/efectos de los fármacos , Animales , Radioisótopos de Carbono , Colágeno/administración & dosificación , Colágeno/efectos de los fármacos , Inyecciones Subcutáneas , Marcaje Isotópico , Masculino , Inhibidores de Proteasas/administración & dosificación , Ratas , Ratas Wistar , Piel/metabolismo , Factores de TiempoRESUMEN
A detailed understanding of the biochemical events leading to the proteolytic excision of the ß-amyloid peptide (A ß) from the amyloid precursor protein (APP) has eluded many researchers. This is largely because the measurement of the various APP processing products is technically challenging owing to their low levels of production in in vitro and in vivo test systems. Sequence analysis of products in cell cultures, cerebrospinal fluid (CSF), and amyloid plaques has been used to predict the major cleavage sites resulting from the ß- and γ-secretase proteolytic activities that release the Aß peptide from APP (1 -3). More routine identification of the secretase activities has relied on the specificity and sensitivity of antibodies raised to the predicted cleavage products and has been impeded by the difficulties associated with the generation of such reagents.
RESUMEN
Members of the PRDM protein family have been shown to play important roles during embryonic development. Previous in vitro and in situ analyses indicated a function of Prdm6 in cells of the vascular system. To reveal physiological functions of Prdm6, we generated conditional Prdm6-deficient mice. Complete deletion of Prdm6 results in embryonic lethality due to cardiovascular defects associated with aberrations in vascular patterning. However, smooth muscle cells could be regularly differentiated from Prdm6-deficient embryonic stem cells and vascular smooth muscle cells were present and proliferated normally in Prdm6-deficient embryos. Conditional deletion of Prdm6 in the smooth muscle cell lineage using a SM22-Cre driver line resulted in perinatal lethality due to hemorrhage in the lungs. We thus identified Prdm6 as a factor that is essential for the physiological control of cardiovascular development.
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Sistema Cardiovascular/embriología , Proteínas Represoras/fisiología , Animales , Secuencia de Bases , Northern Blotting , Southern Blotting , Tipificación del Cuerpo , Diferenciación Celular , Proliferación Celular , Cartilla de ADN , Ratones , Ratones Noqueados , Músculo Liso/citología , Neovascularización Fisiológica , Análisis de Secuencia por Matrices de Oligonucleótidos , Reacción en Cadena de la Polimerasa , Proteínas Represoras/genéticaRESUMEN
BACKGROUND AND PURPOSE: γ-Secretase modulators represent a promising therapeutic approach for Alzheimer's disease (AD) because they selectively decrease amyloid ß 42 (Aß42), a particularly neurotoxic Aß species that accumulates in plaques in the brains of patients with AD. In the present study, we describe the in vitro and in vivo pharmacological properties of a potent novel γ-secretase modulator, 2-(S)-(3,5-bis(4-(trifluoromethyl)phenyl)phenyl)-4-methylpentanoic acid (JNJ-40418677). EXPERIMENTAL APPROACH: The potency and selectivity of JNJ-40418677 for Aß reduction was investigated in human neuroblastoma cells, rat primary neurones and after treatment with single oral doses in non-transgenic mouse brains. To evaluate the effect of JNJ-40418677 on plaque formation, Tg2576 mice were treated from 6 until 13 months of age via the diet. KEY RESULTS: JNJ-40418677 selectively reduced Aß42 secretion in human neuroblastoma cells and rat primary neurones, but it did not inhibit Notch processing or formation of other amyloid precursor protein cleavage products. Oral treatment of non-transgenic mice with JNJ-40418677 resulted in an excellent brain penetration of the compound and a dose- and time-dependent decrease of brain Aß42 levels. Chronic treatment of Tg2576 mice with JNJ-40418677 reduced brain Aß levels, the area occupied by plaques and plaque number in a dose-dependent manner compared with transgenic vehicle-treated mice. CONCLUSIONS AND IMPLICATIONS: JNJ-40418677 selectively decreased Aß42 production, showed an excellent brain penetration after oral administration in mice and lowered brain Aß burden in Tg2576 mice after chronic treatment. JNJ-40418677 therefore warrants further investigation as a potentially effective disease-modifying therapy for AD.
Asunto(s)
Enfermedad de Alzheimer/tratamiento farmacológico , Secretasas de la Proteína Precursora del Amiloide/metabolismo , Ácidos Pentanoicos/uso terapéutico , Placa Amiloide/tratamiento farmacológico , Enfermedad de Alzheimer/metabolismo , Enfermedad de Alzheimer/patología , Péptidos beta-Amiloides/metabolismo , Animales , Encéfalo/efectos de los fármacos , Encéfalo/metabolismo , Encéfalo/patología , Línea Celular Tumoral , Ciclooxigenasa 1/metabolismo , Ciclooxigenasa 2/metabolismo , Femenino , Humanos , Proteínas de la Membrana/metabolismo , Ratones , Ratones Transgénicos , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Ácidos Pentanoicos/farmacocinética , Ácidos Pentanoicos/farmacología , Placa Amiloide/metabolismo , Placa Amiloide/patología , Ratas , Receptores Notch/metabolismoRESUMEN
Translocations involving the immunoglobulin heavy chain locus (IGH@) at chromosome band 14q32 are common in mature B-cell neoplasms, but are rare in B-cell precursor acute lymphoblastic leukemia (BCP-ALL). Here, we report the translocation, t(6;14)(p22;q32), involving IGH@ as a novel recurrent translocation in 13 BCP-ALL patients. Fluorescence in situ hybridization and long-distance inverse polymerase chain reaction (PCR) identified ID4 as the partner gene. Breakpoints were scattered over a 19kb region centromeric of ID4. Quantitative real-time PCR showed up-regulation of ID4 mRNA. All patients had deletions of CDKN2A and PAX5 located on the short arm of chromosome 9, frequently as a result of an isochromosome, i(9)(q10) (9/13, 69%). This study defines a new subgroup of BCP-ALL characterized by ID4 over-expression and CDKN2A and PAX5 deletions. Preliminary survival data suggest that this subgroup may be associated with a good response to therapy.
Asunto(s)
Cadenas Pesadas de Inmunoglobulina/genética , Proteínas Inhibidoras de la Diferenciación/genética , Leucemia de Células B/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Translocación Genética , Adolescente , Adulto , Secuencia de Bases , Niño , Cromosomas Humanos Par 14 , Cromosomas Humanos Par 6 , Cromosomas Humanos Par 9 , Femenino , Eliminación de Gen , Genes p16 , Humanos , Hibridación Fluorescente in Situ , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Factor de Transcripción PAX5/genética , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Integrative genomic and gene-expression analyses have identified amplified oncogenes in B-cell non-Hodgkin lymphoma (B-NHL), but the capability of such technologies to localize tumor suppressor genes within homozygous deletions remains unexplored. Array-based comparative genomic hybridization (CGH) and gene-expression microarray analysis of 48 cell lines derived from patients with different B-NHLs delineated 20 homozygous deletions at 7 chromosome areas, all of which contained tumor suppressor gene targets. Further investigation revealed that only a fraction of primary biopsies presented inactivation of these genes by point mutation or intragenic deletion, but instead some of them were frequently silenced by epigenetic mechanisms. Notably, the pattern of genetic and epigenetic inactivation differed among B-NHL subtypes. Thus, the P53-inducible PIG7/LITAF was silenced by homozygous deletion in primary mediastinal B-cell lymphoma and by promoter hypermethylation in germinal center lymphoma, the proapoptotic BIM gene presented homozygous deletion in mantle cell lymphoma and promoter hypermethylation in Burkitt lymphoma, the proapoptotic BH3-only NOXA was mutated and preferentially silenced in diffuse large B-cell lymphoma, and INK4c/P18 was silenced by biallelic mutation in mantle-cell lymphoma. Our microarray strategy has identified novel candidate tumor suppressor genes inactivated by genetic and epigenetic mechanisms that substantially vary among the B-NHL subtypes.
Asunto(s)
Proteínas Reguladoras de la Apoptosis/genética , Proteínas Portadoras/genética , Inhibidor p18 de las Quinasas Dependientes de la Ciclina/genética , Genes Supresores de Tumor , Proteínas de Homeodominio/genética , Linfoma de Células B/genética , Proteínas de la Membrana/genética , Proteínas Nucleares/genética , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteínas Proto-Oncogénicas/genética , Eliminación de Secuencia , Factores de Transcripción/genética , Proteínas de Transporte Vesicular/genética , Proteínas Adaptadoras Transductoras de Señales , Apoptosis/genética , Proteína 11 Similar a Bcl2 , Biopsia , Línea Celular Tumoral , Mapeo Cromosómico , Cromosomas Humanos/genética , Cromosomas Humanos/ultraestructura , Metilación de ADN , Análisis Mutacional de ADN , ADN de Neoplasias/genética , Epigénesis Genética , Dosificación de Gen , Regulación Neoplásica de la Expresión Génica , Silenciador del Gen , Homocigoto , Humanos , Linfoma de Células B/clasificación , Linfoma de Células B/inmunología , Linfoma de Células B/patología , Hibridación de Ácido Nucleico , Análisis de Secuencia por Matrices de Oligonucleótidos , Mutación Puntual , Regiones Promotoras Genéticas/genética , Proteínas de Unión al ARN , Nexinas de ClasificaciónRESUMEN
CCAAT enhancer-binding protein (CEBP) transcription factors play pivotal roles in proliferation and differentiation, including suppression of myeloid leukemogenesis. Mutations of CEBPA are found in a subset of acute myeloid leukemia (AML) and in some cases of familial AML. Here, using cytogenetics, fluorescence in situ hybridization (FISH), and molecular cloning, we show that 5 CEBP gene family members are targeted by recurrent IGH chromosomal translocations in BCP-ALL. Ten patients with t(8;14)(q11;q32) involved CEBPD on chromosome 8, and 9 patients with t(14;19)(q32;q13) involved CEBPA, while a further patient involved CEBPG, located 71 kb telomeric of CEBPA in chromosome band 19q13; 4 patients with inv(14)(q11q32)/t(14;14)(q11;q32) involved CEBPE and 3 patients with t(14;20)(q32;q13) involved CEBPB. In 16 patients the translocation breakpoints were cloned using long-distance inverse-polymerase chain reaction (LDI-PCR). With the exception of CEBPD breakpoints, which were scattered within a 43-kb region centromeric of CEBPD, translocation breakpoints were clustered immediately 5' or 3' of the involved CEBP gene. Except in 1 patient with t(14;14)(q11;q32), the involved CEBP genes retained germ-line sequences. Quantitative reverse transcription (RT)-PCR showed overexpression of the translocated CEBP gene. Our findings implicate the CEBP gene family as novel oncogenes in BCP-ALL, and suggest opposing functions of CEBP dysregulation in myeloid and lymphoid leukemogenesis.