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1.
Am J Ophthalmol Case Rep ; 25: 101359, 2022 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-35097240

RESUMEN

PURPOSE: To report 3 otherwise healthy patients with Herpes zoster reactivation shortly after administration of a mRNA vaccine against the novel COVID-19 virus. OBSERVATIONS: Patient 1 is a 54 year old who presented with Herpes zoster meningitis complicated by enhancing nodular leptomeningeal lesions of the spinal cord. The subsequent two patients had Herpes zoster ophthalmicus of the cornea (Case 2) and eyelid (Case 3). All three presented within 2 weeks of receiving the Pfizer/BioNTech COVID-19 vaccine. CONCLUSIONS: Herpes zoster may be a side effect of m RNA vaccination against the Sars-CoV2 vaccine and requires further investigation.

2.
Artículo en Inglés | MEDLINE | ID: mdl-29076644

RESUMEN

Immunofluorescence diagnostic systems cost is often dominated by high-sensitivity, low-noise CCD-based cameras that are used to acquire the fluorescence images. In this paper, we investigate the use of low-cost CMOS sensors in a point-of-care immunofluorescence diagnostic application for the detection and discrimination of 4 different serotypes of the Dengue virus in a set of human samples. A 2-phase postprocessing software pipeline is proposed, which consists in a first image-enhancement stage for resolution increasing and segmentation and a second diagnosis stage for the computation of the output concentrations. We present a novel variational coupled model for the joint super-resolution and segmentation stage and an automatic innovative image analysis for the diagnosis purpose. A specially designed forward backward-based numerical algorithm is introduced, and its convergence is proved under mild conditions. We present results on a cheap prototype CMOS camera compared with the results of a more expensive CCD device, for the detection of the Dengue virus with a low-cost OLED light source. The combination of the CMOS sensor and the developed postprocessing software allows to correctly identify the different Dengue serotype using an automatized procedure. The results demonstrate that our diagnostic imaging system enables camera cost reduction up to 99%, at an acceptable diagnostic accuracy, with respect to the reference CCD-based camera system. The correct detection and identification of the Dengue serotypes have been confirmed by standard diagnostic methods (RT-PCR and ELISA).


Asunto(s)
Técnica del Anticuerpo Fluorescente , Sistemas de Atención de Punto , Algoritmos , Humanos , Aumento de la Imagen , Programas Informáticos
3.
N Biotechnol ; 30(5): 475-84, 2013 Jun 25.
Artículo en Inglés | MEDLINE | ID: mdl-23183540

RESUMEN

The analysis of human genetic variability can lead to the comprehension of medical issues and to the development of personalized therapeutic protocols. Single nucleotide polymorphisms, are the most common type of human genetic variation and have been associated to disease development and phenotype forecasting. The recent technologies for DNA sequencing and bioinformatic analysis are now giving the opportunity to develop new diagnostic and prevention approaches also through health promotion protocols. The genetic data management is at the same time underlining technical limitations and old ethical issues.


Asunto(s)
Discusiones Bioéticas , Genética Médica/métodos , Genética Médica/tendencias , Fenotipo , Polimorfismo de Nucleótido Simple , Análisis de Secuencia de ADN , Humanos
4.
Minerva Cardioangiol ; 60(4): 433-7, 2012 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-22858921

RESUMEN

The hepatic artery is the second most common site for aneurysms formation within the splanchnic circulation. Most hepatic artery aneurysms (HAA) are diagnosed incidentally by a computed tomography(CT) scan or a Doppler ultrasonography. We present the case of a HAA diagnosed preoperatively in a 82-year old man, who was treated with an endovascular procedure. An abdominal ultrasonography revealed by chance the presence of a HAA. The abdominal CT scan confirmed an aneurysm of the common hepatic artery, specifically at the origin of the gastroduodenal artery. The gastroduodenal artery was embolized using coils then a heparin-bonded covered stent was deployed into the common hepatic artery to exclude the aneurysm. Final arteriogram documented the regular patency of the stent and the complete exclusion of the aneurysm. No complication occurred and the patient was discharged on the second postoperative day. Six months later, a follow-up with a Duplex scan confirmed the regular patency of the stent, and the patient was in good clinical conditions.


Asunto(s)
Aneurisma/terapia , Anticoagulantes/administración & dosificación , Stents Liberadores de Fármacos , Heparina/administración & dosificación , Arteria Hepática , Anciano de 80 o más Años , Aneurisma/diagnóstico por imagen , Estudios de Seguimiento , Arteria Hepática/diagnóstico por imagen , Humanos , Hallazgos Incidentales , Masculino , Radiografía , Resultado del Tratamiento , Ultrasonografía , Grado de Desobstrucción Vascular
5.
Radiol Med ; 112(8): 1087-99, 2007 Dec.
Artículo en Inglés, Italiano | MEDLINE | ID: mdl-18074200

RESUMEN

The aim of this review is to provide--starting from anatomical, surgical and pathophysiological data--elements for evaluating the status of coronary artery bypass grafts with multidetector computed tomography (CT), taking into consideration the most common conduits used (left and right internal mammary arteries, saphenous vein, radial artery, gastroepiploic artery) and early and late complications (stenosis or obstruction, vasospasm, aneurysms and pseudoaneurysms, malposition). Some of the major problems regarding the examination technique and image analysis are also discussed. Finally, we offer general guidelines for reporting the examination results.


Asunto(s)
Puente de Arteria Coronaria , Complicaciones Posoperatorias/diagnóstico por imagen , Tomografía Computarizada por Rayos X/métodos , Artefactos , Humanos , Imagenología Tridimensional , Interpretación de Imagen Radiográfica Asistida por Computador
6.
Radiol Med ; 112(4): 538-49, 2007 Jun.
Artículo en Inglés, Italiano | MEDLINE | ID: mdl-17563850

RESUMEN

PURPOSE: The study was undertaken to evaluate the 3-year outcome of patients undergoing coronary artery bypass grafting (CABG) involving the use of the radial artery (RA) in comparison with the left internal mammary artery (LIMA) and saphenous vein (SV) grafts by using 16-slice multidetector computed tomography (MDCT). MATERIALS AND METHODS: Fifty-one patients underwent electrocardiogram (ECG)-gated 16-MDCT 32+/-4 months after surgery. A total of 50 LIMA grafts, 55 SV grafts and 51 RA grafts were studied. Approximately 68.6% or RAs were free, 21.5% sequential and 9.8% composite. Grade 0 was defined as complete patency, grade 1 as focal stenosis (>70%) and grade 2 as graft occlusion. The Fisher exact test was used to analyse variables (p<0.05 significant). Concordance between readers for the detection of patency was calculated by the kappa-value. RESULTS: LIMA had the best patency rate (94.0%), followed by SV (83.6%) and RA (74.5%). Regarding RA, the patency rate by territory was 79.4% in the left circumflex coronary artery (LCX), 72.7% in the left anterior descending (LAD) and 50% in the right coronary artery (RCA); the occlusion rate was 20.0% among free grafts, 18.2% among sequential grafts and 20.0% among composite grafts. The kappa-value was 0.86. CONCLUSIONS: Sixteen-slice MDCT scanners enable accurate analysis of CABG status and are a useful noninvasive diagnostic tool for midterm clinical follow-up of patients who have undergone CABG involving the use of RA.


Asunto(s)
Angiografía Coronaria/métodos , Puente de Arteria Coronaria/métodos , Enfermedad de la Arteria Coronaria/cirugía , Arteria Radial/trasplante , Vena Safena/trasplante , Tomografía Computarizada por Rayos X , Anciano , Femenino , Estudios de Seguimiento , Humanos , Anastomosis Interna Mamario-Coronaria , Masculino , Persona de Mediana Edad , Factores de Tiempo
7.
Histol Histopathol ; 22(1): 61-77, 2007 01.
Artículo en Inglés | MEDLINE | ID: mdl-17128412

RESUMEN

In the evolution of colon rectal cancer (CRC) the imbalance between cell proliferation and apoptosis is considered one of the prominent causes of tumor induction and/or progression. In order to establish the role of anti apoptotic proteins in colon cancer development, we studied with immunohistochemical techniques the expression of Survivin in a mouse model of colon carcinogenesis induced by 1,2-dimethyl-hydrazine treatment. In this mouse model Survivin was over-expressed during tumor development, showing a distribution mimicking that described in the correspondent human malignancies. We also correlated Survivin distribution with COX-2 and beta-Catenin expression patterns. The co-localization of COX-2/beta-Catenin/Survivin in the same epithelial cells in tumor samples lends credence to possible in vivo regulatory effects of COX-2 and beta-Catenin on the intracellular Survivin levels in mouse and human colon cancer.


Asunto(s)
Colon/patología , Ciclooxigenasa 2/biosíntesis , Regulación Neoplásica de la Expresión Génica , Regulación de la Expresión Génica , Proteínas Asociadas a Microtúbulos/biosíntesis , Proteínas de Neoplasias/biosíntesis , 1,2-Dimetilhidrazina/farmacología , Animales , Diferenciación Celular , Colon/metabolismo , Modelos Animales de Enfermedad , Células Epiteliales/citología , Humanos , Inmunohistoquímica/métodos , Proteínas Inhibidoras de la Apoptosis , Ratones , Survivin , Distribución Tisular , beta Catenina/biosíntesis
8.
Nature ; 439(7072): 52-4, 2006 Jan 05.
Artículo en Inglés | MEDLINE | ID: mdl-16397493

RESUMEN

Pluto and its satellite, Charon (discovered in 1978; ref. 1), appear to form a double planet, rather than a hierarchical planet/satellite couple. Charon is about half Pluto's size and about one-eighth its mass. The precise radii of Pluto and Charon have remained uncertain, leading to large uncertainties on their densities. Although stellar occultations by Charon are in principle a powerful way of measuring its size, they are rare, as the satellite subtends less than 0.3 microradians (0.06 arcsec) on the sky. One occultation (in 1980) yielded a lower limit of 600 km for the satellite's radius, which was later refined to 601.5 km (ref. 4). Here we report observations from a multi-station stellar occultation by Charon, which we use to derive a radius, R(C) = 603.6 +/- 1.4 km (1sigma), and a density of rho = 1.71 +/- 0.08 g cm(-3). This occultation also provides upper limits of 110 and 15 (3sigma) nanobar for an atmosphere around Charon, assuming respectively a pure nitrogen or pure methane atmosphere.

9.
Minerva Cardioangiol ; 52(1): 55-60, 2004 Feb.
Artículo en Inglés, Italiano | MEDLINE | ID: mdl-14765039

RESUMEN

The anatomic anomalies of the supra-aortic trunks and their branches are comparatively frequent observations. They often remain asymptomatic throughout life although in some cases, especially if nothing is known of them, they may complicate surgical interventions involving that anatomical region. The case of a female patient who, during thyroidectomy, suffered the ligature of her external carotid artery which was taken mistakenly for the thyroid, is reported. Dissection of the area made it possible to identify an anomaly of the vascular structures with a carotid bifurcation which turned out to be very low, a very deep internal carotid artery on the prevertebral fascia and an absent lower thyroid artery. In the postoperative period, the patient was subjected to MR of the neck. This showed an anatomical anomaly of the left carotid bifurcation which was much lower than normal. An attempt was made to reconstruct the external carotid artery with a saphenous vein but unsuccessfully. The incident did not have serious complications for the patient but it might be asked whether, given the low cost and the absolute non-invasiveness of the examination, it might not be worth while to carry out a duplex scan of the supra-aortic trunks before going ahead with any operation in the cervical region, in consideration above all of the dangerous nature of these vascular structures and of the usefulness of studying pathologies that often remain silent until the occurrence of an event which might well be catastrophic.


Asunto(s)
Arteria Carótida Común/anomalías , Tiroidectomía , Arteria Carótida Externa/cirugía , Femenino , Bocio Nodular/cirugía , Humanos , Ligadura , Angiografía por Resonancia Magnética , Persona de Mediana Edad
10.
Minerva Cardioangiol ; 51(1): 79-83, 83-6, 2003 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-12652264

RESUMEN

BACKGROUND: This paper analyses the causes and describes the best care of recurrent varicose veins after internal saphenectomy. METHODS: A series of 19 patients who had previously undergone internal saphenectomy were selected for surgery due to recurrent varices in the lower limbs. Clinical examination and colour duplex sonography were used as the preoperative diagnostic tools in all patients. No patients underwent phlebography. In 17 cases the main source of reflux was an incontinent saphenous stump at the level of the saphenofemoral junction with varicose cross-groin collaterals. In 2 cases recurrence was caused by incontinence of the upper thigh perforating vein. In 1 of these patients the recurrence also involved the district of the small saphenous vein. Groin neovascularisation was detected in 1 patient. RESULTS: All patients underwent groin re-dissections using transversal incisions: in 9 cases, access to the saphenofemoral junction was obtained under or at the same level as the inguinal fold, and in 10 cases using a suprainguinal route. The vertical inguinal incision was never employed. Incompetent perforating veins (thigh or leg) were ligated or sectioned in 11 patients. Ligations and exeresis of communicating veins were executed in all patients. Müller's phlebectomies were performed intra- or postoperatively on collateral varices in practically all cases. Postoperative ambulatory sclerotherapy was necessary in 6 cases. CONCLUSIONS: A correct surgical approach is only assured by diagnostic accuracy coupled with a precise hemodynamic evaluation. Correct management of the postoperative follow-up of varicose vein surgery is also important.


Asunto(s)
Complicaciones Posoperatorias/fisiopatología , Vena Safena/fisiopatología , Várices/fisiopatología , Anciano , Femenino , Humanos , Pierna/irrigación sanguínea , Masculino , Persona de Mediana Edad , Complicaciones Posoperatorias/diagnóstico por imagen , Complicaciones Posoperatorias/cirugía , Recurrencia , Flujo Sanguíneo Regional/fisiología , Vena Safena/diagnóstico por imagen , Vena Safena/cirugía , Ultrasonografía , Várices/diagnóstico por imagen , Várices/cirugía
11.
Hum Exp Toxicol ; 21(1): 7-11, 2002 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-12046726

RESUMEN

Tobacco smoke is involved in the pathogenesis of several diseases regarding different body systems, mainly cardiovascular and respiratory in addition to its local toxic effect in the oral cavity. The noxious effects of smoke compounds justify the high incidence of periodontal diseases, caries, and neoplastic diseases of oral tissues in smokers. Some toxic components of tobacco smoke, unsaturated and saturated aldehydes, could interact with thiol rich compounds, leading to structural and functional modification of these molecules. Previous papers have demonstrated an in vitro significant decrease of some enzymatic activities, both in plasma and in saliva, following external addition of aldehydes or exposure to cigarette smoke (CS). Furthermore, the same studies underlined the protective effect exerted by the addition of glutathione (GSH) against the damaging role of smoke aldehydes. In this study some salivary enzymes (lactic dehydrogenase [LDH], aspartate aminotransferase [AST] and amylase), and total GSH were measured in 20 volunteers smokers, before and just after smoking a single cigarette. All enzymatic activities showed a significant inhibition following a single cigarette, probably due to the interaction between smoke aldehydes and -SH groups of the enzyme molecules. Moreover, the percentage of the enzymatic inhibition showed a negative correlation with the basal level of salivary GSH. Our results emphasize that not only one cigarette is sufficient to impair the salivary enzymatic activities but also strengthen the proposed protective role of GSH against the noxious biochemical effects of CS.


Asunto(s)
Amilasas/antagonistas & inhibidores , Aspartato Aminotransferasas/antagonistas & inhibidores , Glutatión/metabolismo , L-Lactato Deshidrogenasa/antagonistas & inhibidores , Saliva/enzimología , Fumar/efectos adversos , Adulto , Amilasas/metabolismo , Aspartato Aminotransferasas/metabolismo , Femenino , Humanos , L-Lactato Deshidrogenasa/metabolismo , Masculino , Persona de Mediana Edad , Nicotiana
12.
Science ; 288(5473): 2033-5, 2000 Jun 16.
Artículo en Inglés | MEDLINE | ID: mdl-10856214

RESUMEN

Visible and near-infrared spectroscopic observations of the asteroid 1459 Magnya indicate that it has a basaltic surface. Magnya is at 3. 15 astronomical units (AU) from the sun and has no known dynamical link to any family, to any nearby large asteroid, or to asteroid 4 Vesta at 2.36 AU, which is the only other known large basaltic asteroid. We show that the region of the belt around Magnya is densely filled by mean-motion resonances, generating slow orbital diffusion processes and providing a potential mechanism for removing other basaltic fragments that may have been created on the same parent body as Magnya. Magnya may represent a rare surviving fragment from a larger, differentiated planetesimal that was disrupted long ago.

13.
Eur J Biochem ; 267(10): 2939-44, 2000 May.
Artículo en Inglés | MEDLINE | ID: mdl-10806392

RESUMEN

Leptin is an adipocyte-secreted hormone that regulates body weight and exerts effects on hematopoiesis, reproduction, and immunity. The leptin receptor (OBR) shares sequence similarity and signaling capabilities with receptors for cytokines of the ciliary neurotrophic factor (CNTF) family. Our previous finding that CNTF and leptin exert similar anti-obesity effects and activate common neuronal signaling pathways, prompted us to investigate whether leptin may share with CNTF the ability to regulate the expression of specific neuronal genes. To this end, we established a cell line, derived from the murine septal cholinergic neuronal cell line SN-56, which stably expresses OBR. In this cell line, termed SN-56/OBR, leptin induces STAT transcription factor activation and STAT-dependent reporter gene expression in a manner similar to that of CNTF. Furthermore, in SN-56/OBR cells both CNTF and leptin produce changes in neurotransmitter and neuropeptide phenotype characteristic of cholinergic neurons, such as an increase in choline acetyltransferase and vasoactive intestinal polypeptide, and a decrease in neuropeptide Y expression. SN-56/OBR cells thus constitute an interesting new model system to investigate leptin action in cells of central nervous system origin. Possible physiological implications of OBR's intrinsic ability to regulate cholinergic phenotypic markers are discussed.


Asunto(s)
Proteínas Portadoras/metabolismo , Regulación de la Expresión Génica , Neurotransmisores/biosíntesis , Receptores de Superficie Celular , Receptores Colinérgicos/metabolismo , Empalme Alternativo , Animales , Proteínas Portadoras/genética , Proteínas Portadoras/fisiología , Línea Celular , Colina O-Acetiltransferasa/biosíntesis , Factor Neurotrófico Ciliar/metabolismo , Citocinas/farmacología , Relación Dosis-Respuesta a Droga , Regulación hacia Abajo , Genes Reporteros , Leptina/metabolismo , Ratones , Neuronas/metabolismo , Neuropéptido Y/biosíntesis , Fenotipo , ARN/metabolismo , Receptor de Factor Neurotrófico Ciliar/genética , Receptor de Factor Neurotrófico Ciliar/metabolismo , Receptores de Leptina , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transactivadores/metabolismo , Péptido Intestinal Vasoactivo/biosíntesis
14.
CLAO J ; 26(1): 52-3, 2000 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-10656312

RESUMEN

PURPOSE: To present a case of anterior scarring in an epikeratophakia lenticule and its partially successful treatment with the excimer laser. METHODS: Excimer laser phototherapeutic keratectomy (PTK) was used to reduce corneal scarring in an epikeratophakia lenticule. RESULTS: Visual acuity improved after two sessions of PTK. CONCLUSIONS: PTK is an alternative to epikeratophakia lenticule removal.


Asunto(s)
Cicatriz/cirugía , Córnea/patología , Enfermedades de la Córnea/cirugía , Epiqueratofaquia/efectos adversos , Queratectomía Fotorrefractiva , Adulto , Cicatriz/etiología , Cicatriz/patología , Córnea/cirugía , Enfermedades de la Córnea/etiología , Enfermedades de la Córnea/patología , Humanos , Láseres de Excímeros , Masculino , Personal Militar , Miopía/cirugía , Reoperación , Agudeza Visual
15.
IEEE Trans Image Process ; 9(2): 184-9, 2000.
Artículo en Inglés | MEDLINE | ID: mdl-18255385

RESUMEN

In this paper, multiwavelets are considered in the context of image compression and two orthonormal multiwavelet bases are experimented, each used in connection with its proper prefilter. For evaluating the effectiveness of multiwavelet transform for coding images at low bit-rates, an efficient embedded coding of multiwavelet coefficients has been realized. The performance of this multiwavelet-based coder is compared with the results obtained for scalar wavelets.

16.
CLAO J ; 25(4): 197-9, 1999 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-10555732

RESUMEN

PURPOSE: To describe an unusual case of cicatrizing conjunctivitis. METHODS AND RESULTS: A 59 year old male presented in July 1997 with a complaint of intermittent redness and discharge of the right eye. Medical history was significant for a buccal mucosal lesion biopsy taken two years earlier that revealed subepidermal fibrosis. Conjunctival biopsy, performed to rule out ocular cicatricial pemphigoid, yielded a diagnosis of linear IgA dermatosis, an unusual diagnosis not often considered by ophthalmologists. CONCLUSIONS: The diagnosis of cicatrizing conjunctivitis is often overlooked. Its importance as it pertains to the life threatening disease of ocular cicatricial pemphigoid is clear.


Asunto(s)
Inmunoglobulina A/metabolismo , Penfigoide Benigno de la Membrana Mucosa/diagnóstico , Enfermedades Cutáneas Vesiculoampollosas/diagnóstico , Enfermedades Cutáneas Vesiculoampollosas/metabolismo , Adulto , Membrana Basal/metabolismo , Diagnóstico Diferencial , Humanos , Masculino
17.
Ann Thorac Surg ; 67(6): 1815-6, 1999 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-10391311

RESUMEN

Retrograde cerebral perfusion during deep hypothermic circulatory arrest is a technique used largely during operations on the ascending aorta, aortic arch, or both through a median sternotomy. This method is not frequently used for operations performed through a left thoracotomy because of problematic access to the right side of the heart. We propose a technique allowing retrograde cerebral perfusion through a left thoracotomy in a quick, simple, and efficient manner.


Asunto(s)
Encéfalo/irrigación sanguínea , Procedimientos Quirúrgicos Cardíacos , Paro Cardíaco Inducido , Perfusión/métodos , Aorta/cirugía , Humanos , Toracotomía
18.
Proc Natl Acad Sci U S A ; 96(11): 6417-22, 1999 May 25.
Artículo en Inglés | MEDLINE | ID: mdl-10339602

RESUMEN

We show that an electric treatment in the form of high-frequency, low-voltage electric pulses can increase more than 100-fold the production and secretion of a recombinant protein from mouse skeletal muscle. Therapeutical erythopoietin (EPO) levels were achieved in mice with a single injection of as little as 1 microgram of plasmid DNA, and the increase in hematocrit after EPO production was stable and long-lasting. Pharmacological regulation through a tetracycline-inducible promoter allowed regulation of serum EPO and hematocrit levels. Tissue damage after stimulation was transient. The method described thus provides a potentially safe and low-cost treatment for serum protein deficiencies.


Asunto(s)
Eritropoyetina/genética , Técnicas de Transferencia de Gen , Músculo Esquelético/fisiología , Regiones no Traducidas 5'/genética , Animales , Citomegalovirus/genética , Estimulación Eléctrica , Electroporación/métodos , Eritropoyetina/biosíntesis , Eritropoyetina/sangre , Femenino , Regulación de la Expresión Génica , Cinética , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C3H , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Músculo Esquelético/citología , Plásmidos , Regiones Promotoras Genéticas , Proteínas Recombinantes/biosíntesis , Sensibilidad y Especificidad , Factores de Tiempo
19.
J Virol ; 73(3): 2517-26, 1999 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-9971837

RESUMEN

The adeno-associated virus (AAV) is unique in its ability to target viral DNA integration to a defined region of human chromosome 19 (AAVS1). Since AAVS1 sequences are not conserved in a rodent's genome, no animal model is currently available to study AAV-mediated site-specific integration. We describe here the generation of transgenic rats and mice that carry the AAVS1 3.5-kb DNA fragment. To test the response of the transgenic animals to Rep-mediated targeting, primary cultures of mouse fibroblasts, rat hepatocytes, and fibroblasts were infected with wild-type wt AAV. PCR amplification of the inverted terminal repeat (ITR)-AAVS1 junction revealed that the AAV genome integrated into the AAVS1 site in fibroblasts and hepatocytes. Integration in rat fibroblasts was also observed upon transfection of a plasmid containing the rep gene under the control of the p5 and p19 promoters and a dicistronic cassette carrying the green fluorescent protein (GFP) and neomycin (neo) resistance gene between the ITRs of AAV. The localization of the GFP-Neo sequence in the AAVS1 region was determined by Southern blot and FISH analysis. Lastly, AAV genomic DNA integration into the AAVS1 site in vivo was assessed by virus injection into the quadriceps muscle of transgenic rats and mice. Rep-mediated targeting to the AAVS1 site was detected in several injected animals. These results indicate that the transgenic lines are proficient for Rep-mediated targeting. These animals should allow further characterization of the molecular aspects of site-specific integration and testing of the efficacy of targeted integration of AAV recombinant vectors designed for human gene therapy.


Asunto(s)
Dependovirus/genética , Integración Viral , Animales , Animales Modificados Genéticamente , Células Cultivadas , Terapia Genética , Genoma Viral , Masculino , Ratones , Ratas , Ratas Sprague-Dawley
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