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1.
Dev Biol ; 462(1): 101-115, 2020 06 01.
Artículo en Inglés | MEDLINE | ID: mdl-32243888

RESUMEN

Drosophila Larval hematopoiesis takes place at the lymph gland, where myeloid-like progenitors differentiate into Plasmatocytes and Crystal Cells, under regulation of conserved signaling pathways. It has been established that the Notch pathway plays a specific role in Crystal Cell differentiation and maintenance. In mammalian hematopoiesis, the Notch pathway has been proposed to fulfill broader functions, including Hematopoietic Stem Cell maintenance and cell fate decision in progenitors. In this work we describe different roles that Notch plays in the lymph gland. We show that Notch, activated by its ligand Serrate, expressed at the Posterior Signaling Center, is required to restrain Core Progenitor differentiation. We define a novel population of blood cell progenitors that we name Distal Progenitors, where Notch, activated by Serrate expressed in Lineage Specifying Cells at the Medullary Zone/Cortical Zone boundary, regulates a binary decision between Plasmatocyte and Crystal Cell fates. Thus, Notch plays context-specific functions in different blood cell progenitor populations of the Drosophila lymph gland.


Asunto(s)
Células Madre Hematopoyéticas/citología , Ganglios Linfáticos/metabolismo , Receptores Notch/metabolismo , Animales , Células Sanguíneas/citología , Diferenciación Celular/fisiología , Linaje de la Célula , Proteínas de Drosophila/metabolismo , Proteínas de Drosophila/fisiología , Drosophila melanogaster/metabolismo , Hematopoyesis/fisiología , Proteína Jagged-1/metabolismo , Larva/metabolismo , Receptores Notch/fisiología , Transducción de Señal/fisiología
2.
Ann Oncol ; 29(1): 223-229, 2018 01 01.
Artículo en Inglés | MEDLINE | ID: mdl-29045505

RESUMEN

Background: Detection of circulating tumor DNA can be limited due to their relative scarcity in circulation, particularly while patients are actively undergoing therapy. Exosomes provide a vehicle through which cancer-specific material can be enriched from the compendium of circulating non-neoplastic tissue-derived nucleic acids. We carried out a comprehensive profiling of the pancreatic ductal adenocarcinoma (PDAC) exosomal 'surfaceome' in order to identify surface proteins that will render liquid biopsies amenable to cancer-derived exosome enrichment for downstream molecular profiling. Patients and methods: Surface exosomal proteins were profiled in 13 human PDAC and 2 non-neoplastic cell lines by liquid chromatography-mass spectrometry. A total of 173 prospectively collected blood samples from 103 PDAC patients underwent exosome isolation. Droplet digital PCR was used on 74 patients (136 total exosome samples) to determine baseline KRAS mutation call rates while patients were on therapy. PDAC-specific exosome capture was then carried out on additional 29 patients (37 samples) using an antibody cocktail directed against selected proteins, followed by droplet digital PCR analysis. Exosomal DNA in a PDAC patient resistant to therapy were profiled using a molecular barcoded, targeted sequencing panel to determine the utility of enriched nucleic acid material for comprehensive molecular analysis. Results: Proteomic analysis of the exosome 'surfaceome' revealed multiple PDAC-specific biomarker candidates: CLDN4, EPCAM, CD151, LGALS3BP, HIST2H2BE, and HIST2H2BF. KRAS mutations in total exosomes were detected in 44.1% of patients undergoing active therapy compared with 73.0% following exosome capture using the selected biomarkers. Enrichment of exosomal cargo was amenable to molecular profiling, elucidating a putative mechanism of resistance to PARP inhibitor therapy in a patient harboring a BRCA2 mutation. Conclusion: Exosomes provide unique opportunities in the context of liquid biopsies for enrichment of tumor-specific material in circulation. We present a comprehensive surfaceome characterization of PDAC exosomes which allows for capture and molecular profiling of tumor-derived DNA.


Asunto(s)
Carcinoma Ductal Pancreático/diagnóstico , Exosomas/química , Proteínas de Neoplasias/análisis , Neoplasias Pancreáticas/diagnóstico , Biomarcadores de Tumor/sangre , Carcinoma Ductal Pancreático/sangre , Carcinoma Ductal Pancreático/genética , Carcinoma Ductal Pancreático/patología , Línea Celular Tumoral , Cromatografía Liquida , Análisis Mutacional de ADN , Exosomas/metabolismo , Humanos , Biopsia Líquida/métodos , Proteínas de Neoplasias/sangre , Neoplasias Pancreáticas/sangre , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/patología , Medicina de Precisión , Proteómica , Espectrometría de Masas en Tándem
3.
Diabet Med ; 35(10): 1355-1363, 2018 10.
Artículo en Inglés | MEDLINE | ID: mdl-29855077

RESUMEN

AIMS: To investigate trajectories of daily insulin dose requirements and glycaemic control in children, adolescents and young adults with Type 1 diabetes and to identify factors associated with changing insulin needs and deterioration in HbA1c . METHODS: The sample was a dynamic cohort of 635 children, adolescents and young adults with Type 1 diabetes from one centre. Data from clinic visits occurring over 20 years (1993-2013) were extracted from medical records. From age 7-24 years, we evaluated HbA1c and insulin dose according to sex, insulin regimen and weight status. RESULTS: Participants provided a mean ± sd of 10.7±4.3 years of insulin dose data and 12.0±4.6 years of HbA1c data. At first observation, the mean ± sd age was 10.0±2.6 years, diabetes duration was 2.8±2.1 years, insulin dose was 0.8±0.2 units/kg and HbA1c was 74±18 mmol/mol (8.9±1.6%). Insulin dose was higher in girls at ages 8-13 years (P<0.0001 to P<0.01), but higher in boys/young men at ages 16-21 years (P<0.0001 to P=0.04). HbA1c was higher in girls/young women at ages 16-24 years (P<0.0001 to P=0.01). Compared with injection therapy, pump therapy was associated with lower insulin dose at ages 8-24 years (P<0.0001 to P=0.03) and lower HbA1c at ages 8-22 years (P<0.0001 to P=0.005). HbA1c did not differ between overweight/obese and normal weight individuals, but overweight/obese individuals had higher insulin dose at ages 8-13 years (P<0.0001 to P=0.03). CONCLUSIONS: This longitudinal assessment identifies clinically meaningful modifiable (e.g. insulin regimen) and non-modifiable (e.g. sex) factors predictive of insulin requirements and HbA1c levels in young people with Type 1 diabetes; anticipatory insulin adjustments may improve glycaemic control.


Asunto(s)
Glucemia/metabolismo , Diabetes Mellitus Tipo 1/diagnóstico , Diabetes Mellitus Tipo 1/tratamiento farmacológico , Insulina/administración & dosificación , Adolescente , Adulto , Glucemia/efectos de los fármacos , Niño , Estudios de Cohortes , Diabetes Mellitus Tipo 1/sangre , Relación Dosis-Respuesta a Droga , Femenino , Hemoglobina Glucada/efectos de los fármacos , Hemoglobina Glucada/metabolismo , Humanos , Estudios Longitudinales , Masculino , Pronóstico , Adulto Joven
4.
Nature ; 482(7384): 204-7, 2012 Feb 08.
Artículo en Inglés | MEDLINE | ID: mdl-22318604

RESUMEN

The effects of cavity quantum electrodynamics (QED), caused by the interaction of matter and the electromagnetic field in subwavelength resonant structures, have been the subject of intense research in recent years. The generation of coherent radiation by subwavelength resonant structures has attracted considerable interest, not only as a means of exploring the QED effects that emerge at small volume, but also for its potential in applications ranging from on-chip optical communication to ultrahigh-resolution and high-throughput imaging, sensing and spectroscopy. One such strand of research is aimed at developing the 'ultimate' nanolaser: a scalable, low-threshold, efficient source of radiation that operates at room temperature and occupies a small volume on a chip. Different resonators have been proposed for the realization of such a nanolaser--microdisk and photonic bandgap resonators, and, more recently, metallic, metallo-dielectric and plasmonic resonators. But progress towards realizing the ultimate nanolaser has been hindered by the lack of a systematic approach to scaling down the size of the laser cavity without significantly increasing the threshold power required for lasing. Here we describe a family of coaxial nanostructured cavities that potentially solve the resonator scalability challenge by means of their geometry and metal composition. Using these coaxial nanocavities, we demonstrate the smallest room-temperature, continuous-wave telecommunications-frequency laser to date. In addition, by further modifying the design of these coaxial nanocavities, we achieve thresholdless lasing with a broadband gain medium. In addition to enabling laser applications, these nanoscale resonators should provide a powerful platform for the development of other QED devices and metamaterials in which atom-field interactions generate new functionalities.

5.
Gene Ther ; 24(4): 215-223, 2017 04.
Artículo en Inglés | MEDLINE | ID: mdl-28079862

RESUMEN

CLN2 neuronal ceroid lipofuscinosis is a hereditary lysosomal storage disease with primarily neurological signs that results from mutations in TPP1, which encodes the lysosomal enzyme tripeptidyl peptidase-1 (TPP1). Studies using a canine model for this disorder demonstrated that delivery of TPP1 enzyme to the cerebrospinal fluid (CSF) by intracerebroventricular administration of an AAV-TPP1 vector resulted in substantial delays in the onset and progression of neurological signs and prolongation of life span. We hypothesized that the treatment may not deliver therapeutic levels of this protein to tissues outside the central nervous system that also require TPP1 for normal lysosomal function. To test this hypothesis, dogs treated with CSF administration of AAV-TPP1 were evaluated for the development of non-neuronal pathology. Affected treated dogs exhibited progressive cardiac pathology reflected by elevated plasma cardiac troponin-1, impaired cardiac function and development of histopathological myocardial lesions. Progressive increases in the plasma activity levels of alanine aminotransferase and creatine kinase indicated development of pathology in the liver and muscles. The treatment also did not prevent disease-related accumulation of lysosomal storage bodies in the heart or liver. These studies indicate that optimal treatment outcomes for CLN2 disease may require delivery of TPP1 systemically as well as directly to the central nervous system.


Asunto(s)
Aminopeptidasas/genética , Dipeptidil-Peptidasas y Tripeptidil-Peptidasas/genética , Terapia Genética , Enfermedades por Almacenamiento Lisosomal/terapia , Lipofuscinosis Ceroideas Neuronales/terapia , Serina Proteasas/genética , Aminopeptidasas/uso terapéutico , Animales , Dependovirus , Dipeptidil-Peptidasas y Tripeptidil-Peptidasas/uso terapéutico , Modelos Animales de Enfermedad , Perros , Técnicas de Transferencia de Gen , Vectores Genéticos/genética , Vectores Genéticos/uso terapéutico , Humanos , Infusiones Intraventriculares , Enfermedades por Almacenamiento Lisosomal/genética , Lipofuscinosis Ceroideas Neuronales/genética , Neuronas/metabolismo , Neuronas/patología , Serina Proteasas/uso terapéutico , Tripeptidil Peptidasa 1
6.
Ann Oncol ; 28(4): 741-747, 2017 04 01.
Artículo en Inglés | MEDLINE | ID: mdl-28104621

RESUMEN

Background: Exosomes arise from viable cancer cells and may reflect a different biology than circulating cell-free DNA (cfDNA) shed from dying tissues. We compare exosome-derived DNA (exoDNA) to cfDNA in liquid biopsies of patients with pancreatic ductal adenocarcinoma (PDAC). Patients and methods: Patient samples were obtained between 2003 and 2010, with clinically annotated follow up to 2015. Droplet digital PCR was performed on exoDNA and cfDNA for sensitive detection of KRAS mutants at codons 12/13. A cumulative series of 263 individuals were studied, including a discovery cohort of 142 individuals: 68 PDAC patients of all stages; 20 PDAC patients initially staged with localized disease, with blood drawn after resection for curative intent; and 54 age-matched healthy controls. A validation cohort of 121 individuals (39 cancer patients and 82 healthy controls) was studied to validate KRAS detection rates in early-stage PDAC patients. Primary outcome was circulating KRAS status as detected by droplet digital PCR. Secondary outcomes were disease-free and overall survival. Results: KRAS mutations in exoDNA, were identified in 7.4%, 66.7%, 80%, and 85% of age-matched controls, localized, locally advanced, and metastatic PDAC patients, respectively. Comparatively, mutant KRAS cfDNA was detected in 14.8%, 45.5%, 30.8%, and 57.9% of these individuals. Higher exoKRAS MAFs were associated with decreased disease-free survival in patients with localized disease. In the validation cohort, mutant KRAS exoDNA was detected in 43.6% of early-stage PDAC patients and 20% of healthy controls. Conclusions: Exosomes are a distinct source of tumor DNA that may be complementary to other liquid biopsy DNA sources. A higher percentage of patients with localized PDAC exhibited detectable KRAS mutations in exoDNA than previously reported for cfDNA. A substantial minority of healthy samples demonstrated mutant KRAS in circulation, dictating careful consideration and application of liquid biopsy findings, which may limit its utility as a broad cancer-screening method.


Asunto(s)
Carcinoma Ductal Pancreático/genética , ADN de Neoplasias/sangre , Detección Precoz del Cáncer/métodos , Neoplasias Pancreáticas/genética , Proteínas Proto-Oncogénicas p21(ras)/genética , Adulto , Anciano , Biomarcadores de Tumor/sangre , Biomarcadores de Tumor/genética , Carcinoma Ductal Pancreático/sangre , Carcinoma Ductal Pancreático/patología , ADN de Neoplasias/genética , Supervivencia sin Enfermedad , Exosomas/genética , Femenino , Humanos , Estimación de Kaplan-Meier , Masculino , Persona de Mediana Edad , Mutación , Neoplasias Pancreáticas/sangre , Neoplasias Pancreáticas/patología , Modelos de Riesgos Proporcionales , Reacción en Cadena en Tiempo Real de la Polimerasa , Neoplasias Pancreáticas
7.
Epidemiol Infect ; 145(8): 1624-1634, 2017 06.
Artículo en Inglés | MEDLINE | ID: mdl-28290915

RESUMEN

Haiti has the highest human rabies burden in the Western Hemisphere. There is no published literature describing the public's perceptions of rabies in Haiti, information that is critical to developing effective interventions and government policies. We conducted a knowledge, attitudes and practices survey of 550 community members and 116 health professionals in Pétionville, Haiti in 2013 to understand the perception of rabies in these populations. The majority of respondents (85%) knew that dogs were the primary reservoir for rabies, yet only 1% were aware that bats and mongooses could transmit rabies. Animal bites were recognized as a mechanism of rabies transmission by 77% of the population and 76% were aware that the disease could be prevented by vaccination. Of 172 persons reporting a bite, only 37% sought medical treatment. The annual bite incidence rate in respondents was 0·9%. Only 31% of bite victims reported that they started the rabies vaccination series. Only 38% of respondents reported that their dog had been vaccinated against rabies. The majority of medical professionals recognized that dogs were the main reservoir for rabies (98%), but only 28% reported bats and 14% reported mongooses as posing a risk for rabies infection. Bites were reported as a mechanism of rabies transmission by 73% of respondents; exposure to saliva was reported by 20%. Thirty-four percent of medical professionals reported they would wash a bite wound with soap and water and 2·8% specifically mentioned rabies vaccination as a component of post-bite treatment. The majority of healthcare professionals recommended some form of rabies assessment for biting animals; 68·9% recommended a 14-day observation period, 60·4% recommended a veterinary consultation, and 13·2% recommended checking the vaccination status of the animal. Fewer than 15% of healthcare professionals had ever received training on rabies prevention and 77% did not know where to go to procure rabies vaccine for bite victims. Both study populations had a high level of knowledge about the primary reservoir for rabies and the mode of transmission. However, there is a need to improve the level of knowledge regarding the importance of seeking medical care for dog bites and additional training on rabies prevention for healthcare professionals. Distribution channels for rabies vaccines should be evaluated, as the majority of healthcare providers did not know where rabies vaccines could be obtained. Canine rabies vaccination is the primary intervention for rabies control programmes, yet most owned dogs in this population were not vaccinated.


Asunto(s)
Competencia Clínica , Conductas Relacionadas con la Salud , Conocimientos, Actitudes y Práctica en Salud , Personal de Salud/psicología , Rabia/psicología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Haití , Accesibilidad a los Servicios de Salud/estadística & datos numéricos , Humanos , Masculino , Persona de Mediana Edad , Riesgo , Adulto Joven
8.
BJOG ; 124(1): 48-59, 2017 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-27264387

RESUMEN

BACKGROUND: Although pregnant women are considered at high risk for severe influenza disease, comparative studies of maternal influenza and birth outcomes have not been comprehensively summarised. OBJECTIVE: To review comparative studies evaluating maternal influenza disease and birth outcomes. SEARCH STRATEGY: We searched bibliographic databases from inception to December 2014. SELECTION CRITERIA: Studies of preterm birth, small-for-gestational-age (SGA) birth or fetal death, comparing women with and without clinical influenza illness or laboratory-confirmed influenza infection during pregnancy. DATA COLLECTION AND ANALYSIS: Two reviewers independently abstracted data and assessed study quality. MAIN RESULTS: Heterogeneity across 16 studies reporting preterm birth precluded meta-analysis. In a subgroup of the highest-quality studies, two reported significantly increased preterm birth (risk ratios (RR) from 2.4 to 4.0) following severe 2009 pandemic H1N1 (pH1N1) influenza illness, whereas those assessing mild-to-moderate pH1N1 or seasonal influenza found no association. Five studies of SGA birth showed no discernible patterns with respect to influenza disease severity (pooled odds ratio 1.24; 95% CI 0.96-1.59). Two fetal death studies were of sufficient quality and size to permit meaningful interpretation. Both reported an increased risk of fetal death following maternal pH1N1 disease (RR 1.9 for mild-to-moderate disease and 4.2 for severe disease). CONCLUSIONS: Comparative studies of preterm birth, SGA birth and fetal death following maternal influenza disease are limited in number and quality. An association between severe pH1N1 disease and preterm birth and fetal death was reported by several studies; however, these limited data do not permit firm conclusions on the magnitude of any association. TWEETABLE ABSTRACT: Comparative studies are limited in quality but suggest severe pandemic H1N1 influenza increases preterm birth.


Asunto(s)
Recién Nacido Pequeño para la Edad Gestacional , Subtipo H1N1 del Virus de la Influenza A/aislamiento & purificación , Gripe Humana/epidemiología , Complicaciones Infecciosas del Embarazo/epidemiología , Adulto , Femenino , Muerte Fetal/prevención & control , Humanos , Recién Nacido , Gripe Humana/complicaciones , Embarazo , Complicaciones Infecciosas del Embarazo/virología , Resultado del Embarazo , Nacimiento Prematuro/epidemiología , Reino Unido/epidemiología
9.
Avian Pathol ; 46(3): 242-255, 2017 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-27766900

RESUMEN

Infectious bursal disease virus (IBDV) is endemic to most poultry-producing countries worldwide. Immunosuppressive classical and variant IBDV strains endemic to Australia are genetically distinct from other international strains. We report the results of infection experiments with Australian classical strain 06/95 and variant strain 02/95 in SPF chickens. We tested the effects of strain and age of infection on bursal atrophy, viral RNA (vRNA) load in bursa of Fabricius (bursa), spleen, thymus, caecal tonsils, faeces, litter and exhaust dust as determined by real-time reverse transcriptase polymerase chain reaction. The two IBDV strains did not differ in the degree of bursal atrophy induced, lymphoid organ distribution and faecal shedding but variant strain 02/95 induced a greater antibody response to the infection than classical strain 06/95 which was associated with a more rapid decline in IBDV vRNA genome copy number (VCN) in lymphoid organs and faeces. Infection at 14 days of age induced greater bursal atrophy and higher vRNA copy number in lymphoid tissues than infection on the day of hatching, indicating true age susceptibility independent of maternal antibody (Mab) status. The direction of the association between rankings for IBDV vRNA load in bursa and relative bursal weight changed from positive at 3 and 6 days post-infection to negative at 28 days post-infection. Intra-tracheal administration of dust collected from chickens infected with IBDV resulted in successful transmission of IBDV. IBDV vRNA was detected successfully at high levels in the environmental litter and dust samples.


Asunto(s)
Anticuerpos Antivirales/inmunología , Infecciones por Birnaviridae/veterinaria , Pollos/virología , Virus de la Enfermedad Infecciosa de la Bolsa/patogenicidad , Enfermedades de las Aves de Corral/virología , Animales , Infecciones por Birnaviridae/virología , Bolsa de Fabricio/virología , Femenino , Virus de la Enfermedad Infecciosa de la Bolsa/genética , Virus de la Enfermedad Infecciosa de la Bolsa/inmunología , Virus de la Enfermedad Infecciosa de la Bolsa/fisiología , Tejido Linfoide/virología , Masculino , ARN Viral/análisis , Organismos Libres de Patógenos Específicos , Bazo/virología , Distribución Tisular , Carga Viral/veterinaria , Virulencia
10.
Cell Mol Life Sci ; 73(9): 1881-93, 2016 May.
Artículo en Inglés | MEDLINE | ID: mdl-26874685

RESUMEN

Regulation of protein synthesis contributes to maintenance of homeostasis and adaptation to environmental changes. mRNA translation is controlled at various levels including initiation, elongation and termination, through post-transcriptional/translational modifications of components of the protein synthesis machinery. Recently, protein and RNA hydroxylation have emerged as important enzymatic modifications of tRNAs, elongation and termination factors, as well as ribosomal proteins. These modifications enable a correct STOP codon recognition, ensuring translational fidelity. Recent studies are starting to show that STOP codon read-through is related to the ability of the cell to cope with different types of stress, such as oxidative and chemical insults, while correlations between defects in hydroxylation of protein synthesis components and STOP codon read-through are beginning to emerge. In this review we will discuss our current knowledge of protein synthesis regulation through hydroxylation of components of the translation machinery, with special focus on STOP codon recognition. We speculate on the possibility that programmed STOP codon read-through, modulated by hydroxylation of components of the protein synthesis machinery, is part of a concerted cellular response to stress.


Asunto(s)
Estrés Oxidativo , Bacterias/metabolismo , Codón de Terminación , Dioxigenasas/metabolismo , Humanos , Hidroxilación , Procesamiento Proteico-Postraduccional , ARN de Transferencia/genética , ARN de Transferencia/metabolismo , Proteínas Ribosómicas/metabolismo
11.
J Vet Pharmacol Ther ; 40(6): e1-e10, 2017 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-28649788

RESUMEN

Mycophenolate mofetil (MMF) is recommended as an alternative/complementary immunosuppressant. Pharmacokinetic and dynamic effects of MMF are unknown in young-aged dogs. We investigated the pharmacokinetics and pharmacodynamics of single oral dose MMF metabolite, mycophenolic acid (MPA), in healthy juvenile dogs purpose-bred for the tripeptidyl peptidase 1 gene (TPP1) mutation. The dogs were heterozygous for the mutation (nonaffected carriers). Six dogs received 13 mg/kg oral MMF and two placebo. Pharmacokinetic parameters derived from plasma MPA were evaluated. Whole-blood mitogen-stimulated T-cell proliferation was determined using a flow cytometric assay. Plasma MPA Cmax (mean ± SD, 9.33 ± 7.04 µg/ml) occurred at <1 hr. The AUC0-∞ (mean ± SD, 12.84±6.62 hr*µg/ml), MRTinf (mean ± SD, 11.09 ± 9.63 min), T1/2 (harmonic mean ± PseudoSD 5.50 ± 3.80 min), and k/d (mean ± SD, 0.002 ± 0.001 1/min). Significant differences could not be detected between % inhibition of proliferating CD5+ T lymphocytes at any time point (p = .380). No relationship was observed between MPA concentration and % inhibition of proliferating CD5+ T lymphocytes (R = .148, p = .324). Pharmacodynamics do not support the use of MMF in juvenile dogs at the administered dose based on existing therapeutic targets.


Asunto(s)
Inmunosupresores/farmacocinética , Ácido Micofenólico/farmacocinética , Administración Oral , Animales , Antígenos CD5/inmunología , Perros , Femenino , Citometría de Flujo/veterinaria , Inmunosupresores/administración & dosificación , Inmunosupresores/farmacología , Activación de Linfocitos/efectos de los fármacos , Masculino , Ácido Micofenólico/administración & dosificación , Ácido Micofenólico/farmacología , Linfocitos T/efectos de los fármacos , Linfocitos T/inmunología
12.
Gene Ther ; 23(2): 151-7, 2016 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-26461176

RESUMEN

The S100A1 gene is a promising target enhancing contractility and survival post myocardial infarction (MI). Achieving sufficient gene delivery within safety limits is a major translational problem. This proof of concept study evaluates viral mediated S100A1 overexpression featuring a novel liquid jet delivery (LJ) method. Twenty-four rats after successful MI were divided into three groups (n = 8 ea.): saline control (SA); ssAAV9.S100A1 (SS) delivery; and scAAV9.S100A1 (SC) delivery (both 1.2 × 10¹¹ viral particles). For each post MI rat, the LJ device fired three separate 100 µl injections into the myocardium. Following 10 weeks, all rats were evaluated with echocardiography, quantitative PCR (qPCR) and overall S100A1 and CD38 immune protein. At 10 weeks all groups demonstrated a functional decline from baseline, but the S100A1 therapy groups displayed preserved left ventricular function with significantly higher ejection fraction %; SS group (60 ± 3) and SC group (57 ± 4) versus saline (46 ± 3), P < 0.05. Heart qPCR testing showed robust S100A1 in the SS (10,147 ± 3993) and SC (35,155 ± 5808) copies per 100 ng DNA, while off-target liver detection was lower in both SS (40 ± 40), SC (34,841 ± 3164), respectively. Cardiac S100A1 protein expression was (4.3 ± 0.2) and (6.1 ± 0.3) fold higher than controls in the SS and SC groups, respectively, P < 0.05.


Asunto(s)
Técnicas de Transferencia de Gen , Terapia Genética , Infarto del Miocardio/terapia , Proteínas S100/genética , Animales , Dependovirus/genética , Vectores Genéticos , Masculino , Miocardio/metabolismo , Ratas , Ratas Sprague-Dawley , Proteínas S100/biosíntesis , Función Ventricular Izquierda
13.
Ann Oncol ; 27(4): 635-41, 2016 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-26681674

RESUMEN

BACKGROUND: The ability to perform comprehensive profiling of cancers at high resolution is essential for precision medicine. Liquid biopsies using shed exosomes provide high-quality nucleic acids to obtain molecular characterization, which may be especially useful for visceral cancers that are not amenable to routine biopsies. PATIENTS AND METHODS: We isolated shed exosomes in biofluids from three patients with pancreaticobiliary cancers (two pancreatic, one ampullary). We performed comprehensive profiling of exoDNA and exoRNA by whole genome, exome and transcriptome sequencing using the Illumina HiSeq 2500 sequencer. We assessed the feasibility of calling copy number events, detecting mutational signatures and identifying potentially actionable mutations in exoDNA sequencing data, as well as expressed point mutations and gene fusions in exoRNA sequencing data. RESULTS: Whole-exome sequencing resulted in 95%-99% of the target regions covered at a mean depth of 133-490×. Genome-wide copy number profiles, and high estimates of tumor fractions (ranging from 56% to 82%), suggest robust representation of the tumor DNA within the shed exosomal compartment. Multiple actionable mutations, including alterations in NOTCH1 and BRCA2, were found in patient exoDNA samples. Further, RNA sequencing of shed exosomes identified the presence of expressed fusion genes, representing an avenue for elucidation of tumor neoantigens. CONCLUSIONS: We have demonstrated high-resolution profiling of the genomic and transcriptomic landscapes of visceral cancers. A wide range of cancer-derived biomarkers could be detected within the nucleic acid cargo of shed exosomes, including copy number profiles, point mutations, insertions, deletions, gene fusions and mutational signatures. Liquid biopsies using shed exosomes has the potential to be used as a clinical tool for cancer diagnosis, therapeutic stratification and treatment monitoring, precluding the need for direct tumor sampling.


Asunto(s)
Biomarcadores de Tumor/genética , Proteínas de Neoplasias/genética , Neoplasias Pancreáticas/genética , Anciano , Biomarcadores de Tumor/biosíntesis , Exoma/genética , Exosomas/genética , Femenino , Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Masculino , Persona de Mediana Edad , Mutación , Proteínas de Neoplasias/biosíntesis , Neoplasias Pancreáticas/patología
14.
Ann Surg Oncol ; 23(3): 1035, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-26542586

RESUMEN

BACKGROUND: Organ-sparing pancreatic resection is important in prophylactic surgery for cystic neoplasms. There is controversy regarding the optimal surgical approach for pancreatic lesions in the neck or proximal body of the pancreas. Central compared with distal pancreatectomy is technically more challenging, but preserves more functional pancreatic tissue. Because of the prophylactic nature of the surgery and long survival of patients with benign and borderline malignant lesions, surgeons need to stratify greater importance to surgical morbidity and sparing pancreatic parenchyma. PATIENT: The patient is a 59-year-old active woman with a symptomatic cystic neoplasm of the pancreas exhibiting high-risk imaging features. The cyst of 2.2 × 1.8 cm in the body of the pancreas was impinging on the portal venous confluence. TECHNIQUE: The patient was positioned in the French Position, the lesser sac was opened, and the pancreatic body exposed. A retropancreatic tunnel was created with staple division of the neck. The body was mobilized off the portal vein and splenic vessels transected. A retrogastric pancreaticogastrostomy was sewn through an anterior gastrotomy. The stent was delivered past the pylorus to decrease pancreatic enzymatic activation. Pathology demonstrated a mixed predominantly branch duct IPMN with multifocal high grade dysplasia and PanIN3. CONCLUSIONS: Laparoscopic ultrasound helps in defining cyst borders, and minimal blood loss optimizes visualization during the dissection. A minimally invasive pancreaticogastrostomy created through an anterior gastrotomy is technically feasible and safe. This approach can minimize the morbidity of prophylactic pancreatic surgery for patients with cystic neoplasms. Nevertheless, it should not compromise safety, oncologic completeness, or an organ-sparing approach.


Asunto(s)
Gastrostomía/métodos , Laparoscopía/métodos , Pancreatectomía/métodos , Quiste Pancreático/cirugía , Neoplasias Pancreáticas/cirugía , Femenino , Humanos , Persona de Mediana Edad , Pronóstico , Factores de Riesgo
15.
Br J Surg ; 103(3): 179-91, 2016 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-26663252

RESUMEN

BACKGROUND: Pancreatic ductal adenocarcinoma has a poor prognosis without surgery. No standard treatment has yet been accepted for patients with portal-superior mesenteric vein (PV-SMV) infiltration. The present meta-analysis aimed to compare the results of pancreatic resection with PV-SMV resection for suspected infiltration with the results of surgery without PV-SMV resection. METHODS: A systematic search was performed of PubMed, Embase and the Cochrane Library in accordance with PRISMA guidelines from the time of inception to 2013. The inclusion criteria were comparative studies including patients who underwent pancreatic resection with or without PV-SMV resection. One, 3- and 5-year survival were the primary outcomes. RESULTS: Twenty-seven studies were identified involving a total of 9005 patients (1587 in PV-SMV resection group). Patients undergoing PV-SMV resection had an increased risk of postoperative mortality (risk difference (RD) 0.01, 95 per cent c.i. 0.00 to 0.03; P = 0.2) and of R1/R2 resection (RD 0.09, 0.06 to 0.13; P < 0.001) compared with those undergoing standard surgery. One-, 3- and 5-year survival were worse in the PV-SMV resection group: hazard ratio 1.23 (95 per cent c.i. 1.07 to 1.43; P = 0.005), 1.48 (1.14 to 1.91; P = 0.004) and 3.18 (1.95 to 5.19; P < 0.001) respectively. Median overall survival was 14.3 months for patients undergoing pancreatic resection with PV-SMV resection and 19.5 months for those without vein resection (P = 0.063). Neoadjuvant therapies recently showed promising results. CONCLUSION: This meta-analysis showed increased postoperative mortality, higher rates of non-radical surgery and worse survival after pancreatic resection with PV-SMV resection. This may be related to more advanced disease in this group.


Asunto(s)
Carcinoma Ductal Pancreático/cirugía , Venas Mesentéricas/cirugía , Neoplasias Pancreáticas/cirugía , Pancreaticoduodenectomía/métodos , Vena Porta/cirugía , Humanos , Resultado del Tratamiento
17.
Mol Genet Metab ; 115(2-3): 101-9, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-25934231

RESUMEN

We studied a recessive, progressive neurodegenerative disease occurring in Golden Retriever siblings with an onset of signs at 15 months of age. As the disease progressed these signs included ataxia, anxiety, pacing and circling, tremors, aggression, visual impairment and localized and generalized seizures. A whole genome sequence, generated with DNA from one affected dog, contained a plausibly causal homozygous mutation: CLN5:c.934_935delAG. This mutation was predicted to produce a frameshift and premature termination codon and encode a protein variant, CLN5:p.E312Vfs*6, which would lack 39 C-terminal amino acids. Eighteen DNA samples from the Golden Retriever family members were genotyped at CLN5:c.934_935delAG. Three clinically affected dogs were homozygous for the deletion allele; whereas, the clinically normal family members were either heterozygotes (n = 11) or homozygous for the reference allele (n = 4). Among archived Golden Retrievers DNA samples with incomplete clinical records that were also genotyped at the CLN5:c.934_935delAG variant, 1053 of 1062 were homozygous for the reference allele, 8 were heterozygotes and one was a deletion-allele homozygote. When contacted, the owner of this homozygote indicated that their dog had been euthanized because of a neurologic disease that progressed similarly to that of the affected Golden Retriever siblings. We have collected and stored semen from a heterozygous Golden Retriever, thereby preserving an opportunity for us or others to establish a colony of CLN5-deficient dogs.


Asunto(s)
Enfermedades de los Perros/genética , Mutación del Sistema de Lectura , Proteínas de la Membrana/genética , Lipofuscinosis Ceroideas Neuronales/veterinaria , Eliminación de Secuencia , Animales , Secuencia de Bases , Perros , Homocigoto , Lipofuscinosis Ceroideas Neuronales/genética , Análisis de Secuencia de ADN
18.
Diabet Med ; 32(9): 1227-31, 2015 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-25655562

RESUMEN

AIMS: To develop and validate the Diabetes Family Impact Scale, a scale to measure the impact of diabetes on families. METHODS: The Diabetes Family Impact Scale was developed by an iterative process, with input from multidisciplinary diabetes providers and parents of children with Type 1 diabetes. The psychometric properties of the Diabetes Family Impact Scale were assessed in parents of children with Type 1 diabetes. This assessment included internal consistency, convergent validity and exploratory factor analysis. RESULTS: Parents (n = 148) of children (mean ± sd age 12.9 ± 3.3 years) with Type 1 diabetes (mean ± sd duration 6.2 ± 3.6 years) completed the 15-item Diabetes Family Impact Scale. After eliminating one item, the 14-item measure demonstrated good internal consistency (Cronbach's α  =  0.84). Correlations between the Diabetes Family Impact Scale and measures of parent diabetes burden (r = 0.48, P < 0.0001), stressful life events (r = 0.28, P = 0.0007), and child's quality of life (r = -0.52 and -0.54, P < 0.0001 for generic and diabetes-specific quality of life, respectively) supported the convergent validity of the instrument. Factor analysis identified four factors corresponding to the four survey domains (school, work, finances and family well-being). CONCLUSIONS: The Diabetes Family Impact Scale measures diabetes-specific family impacts with good internal consistency and convergent validity and may be a useful tool in clinical and research settings.


Asunto(s)
Diabetes Mellitus Tipo 1/psicología , Salud de la Familia , Encuestas y Cuestionarios/normas , Adolescente , Niño , Costo de Enfermedad , Femenino , Humanos , Masculino , Psicometría
19.
Eur J Cancer Care (Engl) ; 24(1): 28-38, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-24467411

RESUMEN

Patient participation in cancer clinical trials is low. Little is known about attitudinal barriers to participation, particularly among patients who may be offered a trial during an imminent initial oncology consult. The aims of the present study were to confirm the presence of proposed subscales of a recently developed cancer clinical trial attitudinal barriers measure, describe the most common cancer clinical trials attitudinal barriers, and evaluate socio-demographic, medical and financial factors associated with attitudinal barriers. A total of 1256 patients completed a survey assessing demographic factors, perceived financial burden, prior trial participation and attitudinal barriers to clinical trials participation. Results of a factor analysis did not confirm the presence of the proposed four attitudinal barriers subscale/factors. Rather, a single factor represented the best fit to the data. The most highly-rated barriers were fear of side-effects, worry about health insurance and efficacy concerns. Results suggested that less educated patients, patients with non-metastatic disease, patients with no previous oncology clinical trial participation, and patients reporting greater perceived financial burden from cancer care were associated with higher barriers. These patients may need extra attention in terms of decisional support. Overall, patients with fewer personal resources (education, financial issues) report more attitudinal barriers and should be targeted for additional decisional support.


Asunto(s)
Toma de Decisiones , Conocimientos, Actitudes y Práctica en Salud , Neoplasias/psicología , Participación del Paciente/psicología , Anciano , Ensayos Clínicos como Asunto , Estudios Transversales , Análisis Factorial , Femenino , Humanos , Masculino , Persona de Mediana Edad , Neoplasias/terapia , Participación del Paciente/economía , Participación del Paciente/estadística & datos numéricos , Derivación y Consulta , Encuestas y Cuestionarios
20.
Vet Pathol ; 51(4): 796-804, 2014 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-24026940

RESUMEN

Three young domestic shorthair cats were presented for necropsy with similar histories of slowly progressive visual dysfunction and neurologic deficits. Macroscopic examination of each cat revealed cerebral and cerebellar atrophy, dilated lateral ventricles, and slight brown discoloration of the gray matter. Histologically, there was bilateral loss of neurons within the limbic, motor, somatosensory, visual, and, to a lesser extent, vestibular systems with extensive astrogliosis in the affected regions of all 3 cases. Many remaining neurons and glial cells throughout the entire central nervous system were distended by pale yellow to eosinophilic, autofluorescent cytoplasmic inclusions with ultrastructural appearances typical of neuronal ceroid-lipofuscinoses (NCLs). Differences in clinical presentation and neurological lesions suggest that the 3 cats may have had different variants of NCL. Molecular genetic characterization in the 1 cat from which DNA was available did not reveal any plausible disease-causing mutations of the CLN1 (PPT1), CLN3, CLN5, CLN8, and CLN10 (CTSD) genes. Further investigations will be required to identify the mutations responsible for NCLs in cats.


Asunto(s)
Enfermedades de los Gatos/patología , Sistema Nervioso/patología , Lipofuscinosis Ceroideas Neuronales/veterinaria , Animales , Atrofia/patología , Atrofia/veterinaria , Gatos , Análisis Mutacional de ADN/veterinaria , Resultado Fatal , Técnicas Histológicas/veterinaria , Inmunohistoquímica/veterinaria , Minnesota , Lipofuscinosis Ceroideas Neuronales/patología
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