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1.
Cir Pediatr ; 31(2): 99-103, 2018 Apr 20.
Artigo em Espanhol | MEDLINE | ID: mdl-29978963

RESUMO

OBJECTIVE: To compare postoperative follow up in patients older and younger than 12 months who underwent surgical treatment of ureteropelvic junction obstruction (UJO). MATERIAL AND METHODS: Retrospective study of 77 patients, 78 kidney units, intervened from UJO (2007-2014). We analyzed epidemiological, clinical, echographic, and pre and postoperative renogram variables, outcomes and complications. We divided the patients into 2 groups according to age: group A ≤ 12 months and group B > 12 months, comparing the results by statistical analysis, considering p < 0.05 statistically significant. RESULTS: Group A: 38 patients, 26 males (68.4%), one bilateral UJO and 22 rights (57.9%), 36 prenatal diagnoses (92.3%) and mean age of intervention 5.28 months [range 0.24 -11,28]. We performed 9 minilumbotomies, 29 assisted by retroperitoneoscopy (ARP) and 1 pneumatic dilation (PD). Group B: 39 patients, 26 males (66.7%), 10 rights (25.64%), 19 prenatal diagnoses (48.7%) and mean age 6.13 years [range 1.13-14.52]. 15 minilumbotomies, 20 ARP, 3 laparoscopic and 1 PD. Preoperative mean renal function (MRF) of group A: 35.9 ± 13.4 [range 8-57] vs. 39.74 ± 13.91 [range 9-57] in group B (p = 0.347). Postoperative MRF 43.29 ± 18.2 [range 12-100] group A and 39.41 ± 12.89 [range 11-54] group B (p = 0.464). Group A and B: 11 and 8 complications, respectively (p = 0.429). We did not find statistically significant differences in the mean preoperative anteroposterior diameter (DAP) between both groups (p = 0.313). We compared DAP at 3, 6, 12, 24 and 48 postoperative months, observing a greater reduction of DAP from group A compared to B; however, we found only statistically significant differences in DAP at 3 months postoperatively (p = 0.047). CONCLUSION: Renal DAP is reduced postoperatively more in patients younger than 1 year. Moreover, an improvement of the DRF after pieloplasty can be observed despite not being statistically significant.


OBJETIVO: Comparar la evolución postquirúrgica en pacientes mayores y menores de 12 meses intervenidos de estenosis pieloureteral (EPU). MATERIAL Y METODOS: Estudio retrospectivo de 77 pacientes, 78 unidades renales, intervenidos por EPU (2007-2014). Analizamos variables epidemiológicas, clínicas, ecográficas y de renogramas pre y postoperatorios, resultados y complicaciones. Dividimos a los pacientes en 2 grupos según la edad: grupo A ≤ 12 meses y grupo B > 12 meses, comparando los resultados mediante análisis estadísticos (p < 0,05 estadísticamente significativo). RESULTADOS: Grupo A: 38 pacientes, 26 varones (68,4%), una EPU bilateral y 22 derechas (57,9%), 36 diagnósticos prenatales (92,3%) y edad media de intervención 5,28 meses [rango 0,24-11,28]. Realizamos 9 minilumbotomías, 29 asistidas por retroperitoneoscopia (ARP) y una dilatación neumática (DN). Grupo B: 39 pacientes, 26 varones (66,7%), 10 derechas (25,64%), 19 diagnósticos prenatales (48,7%) y edad media 6,13 años [rango 1,13-14,52]. Realizamos 15 minilumbotomías, 20 ARP, 3 laparoscópicas y 1 DN. Función renal diferencial media (FRDM) preoperatoria del grupo A: 35,9 ± 13,4 [rango 8-57] vs. 39,74 ± 13,91 [rango 9-57] grupo B (p = 0,347). FRDM postoperatoria 43,29 ± 18,2 [rango 12-100] grupo A y 39,41 ± 12,89 [rango 11-54] grupo B (p = 0,464). Grupos A y B: 11 y 8 complicaciones, respectivamente (p = 0,429). No encontramos diferencias estadísticamente significativas en la media del diámetro anteroposterior (DAP) de la pelvis preoperatoria entre ambos grupos (p = 0,313). Comparamos los DAP a los 3, 6, 12, 24 y 48 meses postoperatorios, observando una reducción mayor del DAP del grupo A frente al B, sin embargo, solo encontramos diferencias estadísticamente significativas en el DAP a los 3 meses postoperatorios (p = 0,047). CONCLUSION: El DAP de la pelvis renal se reduce más en los pacientes menores de 1 año a los 3 meses postoperatorios. Además, podemos observar una evidente mejoría de la FRDM tras la pieloplastia a pesar de no encontrar diferencias estadísticamente significativas.


Assuntos
Pelve Renal/cirurgia , Laparoscopia/métodos , Diagnóstico Pré-Natal/métodos , Obstrução Ureteral/cirurgia , Adolescente , Fatores Etários , Criança , Pré-Escolar , Feminino , Seguimentos , Humanos , Lactente , Recém-Nascido , Testes de Função Renal , Pelve Renal/patologia , Masculino , Período Pós-Operatório , Gravidez , Estudos Retrospectivos , Obstrução Ureteral/diagnóstico
2.
J Adv Nurs ; 71(6): 1417-34, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25250659

RESUMO

AIM: To describe the associations between positive and negative reactions of informal caregivers of people with dementia and health outcomes across eight European Countries. BACKGROUND: Caring for someone with dementia may have implications for the caregiver's own health and for the care recipient. These consequences could be associated with caregivers' reactions to the process of care. DESIGN: Association study based on cross-sectional data. METHODS: Participants were people with dementia and their informal caregivers living at home or in long-term care institutions. Data were collected between November 2010-April 2012 using the Caregiver Reaction Assessment (with dimensions of self-esteem, lack of family support, financial problems, disrupted schedule and health problems) and associations were sought with informal caregiver burden, quality of life and psychological well-being and with dementia sufferers' neuropsychiatric symptoms, comorbidity and dependency in activities of daily living using correlation coefficients. RESULTS: Data from 2014 participants were used. Variability across countries was noted, as well as differences between care at home and in long-term care institutions. In general, self-esteem and lack of family support correlated with caregiver burden and psychological well-being. Associations were also found between disrupted schedule and caregiver burden, psychological well-being and quality of life. Health problems were clearly associated with caregiver burden, psychological well-being and quality of life. CONCLUSION: Study results support links between the reactions of informal caregivers of people with dementia and health outcomes. These may have implications in terms of how services are addressed.


Assuntos
Cuidadores/psicologia , Demência/enfermagem , Estudos Transversais , Europa (Continente) , Serviços de Assistência Domiciliar , Humanos , Qualidade de Vida , Resultado do Tratamento
3.
Cir Pediatr ; 35(2): 91-93, 2022 Apr 01.
Artigo em Inglês, Espanhol | MEDLINE | ID: mdl-35485758

RESUMO

INTRODUCTION: Urethrorrhagia is an infrequent sign in childhood. It should be distinguished from hematuria, since they have a different etiology. CLINICAL CASE: 11-year-old male patient with significant urethrorrhagia. Urinary sediment analysis: red blood cells++. Pelvic ultrasonography: fusiform anechoic image in the corpus spongiosum of the penile root. Retrograde urethrogram: normal anterior urethra, extraluminal contrast passage in the ventral aspect of the bulbar urethra. Cystoscopy: no pathological findings in the urethra or the bladder. Control retrograde urethrogram: cystic dilatation of Cowper's gland duct; Maizels' type 3 perforated syringocele. DISCUSSION: Cowper's syringocele is a rare pathology. It can occur at any stage of childhood in the form of urinary infection, obstructive voiding symptoms, or urethrorrhagia. Urethrogram is key for diagnostic purposes, since most Cowper's syringoceles are detected following urethrogram or cystoscopy. Cases with functional repercussions for the urinary system require surgical treatment. Otherwise, a wait-and-see approach is feasible.


INTRODUCCION: La uretrorragia es un signo infrecuente en la infancia que debe distinguirse de la hematuria dada la diferente etiología de las mismas. CASO CLINICO: Varón de 11 años con uretrorragia franca. Sedimento urinario: hematíes++. Ecografía pélvica: imagen anecoica fusiforme en cuerpo esponjoso de raíz peneana. Uretrografía retrógrada: uretra anterior normal, paso de contraste extraluminal ventral en uretra bulbar. Cistoscopia: sin hallazgos patológicos en uretra ni vejiga. Uretrografía retrógrada de control: dilatación quística del conducto de las glándulas de Cowper; siringocele perforado tipo 3 de Maizels. COMENTARIOS: El siringocele de Cowper es una patología infrecuente que puede debutar en cualquier momento de la infancia como infección urinaria, síntomas miccionales obstructivos o uretrorragia. La uretrografía es fundamental en su diagnóstico ya que la mayoría se objetivan por este medio o cistoscopia. Los casos con repercusión funcional del sistema urinario requieren tratamiento quirúrgico. En caso contrario podrá realizarse actitud expectante.


Assuntos
Cirurgiões , Doenças Uretrais , Glândulas Bulbouretrais/patologia , Criança , Feminino , Humanos , Masculino , Radiografia , Uretra/diagnóstico por imagem , Uretra/cirurgia , Doenças Uretrais/diagnóstico por imagem , Doenças Uretrais/cirurgia
4.
Mol Cell Biol ; 13(1): 578-87, 1993 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-8417354

RESUMO

For the majority of mRNA encoding eukaryotic transcription units, there is little or no knowledge of the elements responsible for transcription termination or how they may interact with RNA polymerase. In this report, we have used recombinant adenovirus reporter vectors to characterize the mouse beta maj globin sequence elements that cause transcription termination. Within the globin 3' termination region, we have identified at least three sequence elements which induce significant levels of transcription termination (> 50%). The smallest functionally active element (64% termination) is 69 bp in length. The natural arrangement of these elements results in a cumulative termination which is greater than 90%. Recognition of the termination elements by RNA polymerase II depends on the presence of a functional poly(A) signal sequence. We demonstrate that efficient transcription termination depends on appropriate spacing between the poly(A) signal sequence and the termination element.


Assuntos
Globinas/genética , RNA Mensageiro/genética , Regiões Terminadoras Genéticas , Animais , Sequência de Bases , Células HeLa , Humanos , Técnicas In Vitro , Camundongos , Dados de Sequência Molecular , Poli A/metabolismo , RNA Polimerase II/metabolismo , Processamento Pós-Transcricional do RNA , Sequências Reguladoras de Ácido Nucleico , Deleção de Sequência
5.
Arch Intern Med ; 138(8): 1247-50, 1978 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-209756

RESUMO

Cytomegaloviurs (CMV) infections are a recognized problem in the first six months after renal transplantation. Studies have suggested that primary infections produce symptomatic disease, whereas reactivation infections are usually asymptomatic. Two patients are described who developed fatal CMV infections in the second year after transplantation. Both patients had typical CMV disease with fever, pneumonitis, and hepatitis. Results of serologic studies in one patient were characteristic of primary infection, with seroconversion at the time of disease and appearance of specific IgM antibodies. The other patient had a similar antibody response at the time of his illness, but serial antibody tests showed that he had had a transient seroconversion earlier, in the second month after transplanation, that was not associated with clinical symptoms. These patients indicate that CMV infection must be considered in the differential diagnosis of serious febrile illnesses even in the late posttransplantation period and may occur either as the result of primary or reactivation infection. Serologic studies at the time of illness may not allow distinction between the types of infection.


Assuntos
Infecções por Citomegalovirus/etiologia , Transplante de Rim , Complicações Pós-Operatórias , Adulto , Anticorpos Antivirais/análise , Citomegalovirus/imunologia , Infecções por Citomegalovirus/imunologia , Humanos , Imunoglobulina M/análise , Masculino , Pessoa de Meia-Idade , Transplante Homólogo
6.
Arch Neurol ; 37(4): 239-40, 1980 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-6244805

RESUMO

A 20-year-old man with Hodgkin's disease experienced ophthalmic zoster with dissemination and CNS involvement. At autopsy, he was found to have granulomatous angiitis involving the basilar artery, and electron microscopy revealed virus-like particles in the outer layers of the vessel walls, but not the endothelium. This suggests that granulomatous angiitis of the CNS in varicella-zoster infections results from direct viral invasion of blood vessels, perhaps by contiguous spread from cranial nerves.


Assuntos
Arterite/patologia , Artéria Basilar/patologia , Varicela/patologia , Adulto , Granuloma/patologia , Herpesvirus Humano 3/ultraestrutura , Doença de Hodgkin/patologia , Humanos , Masculino , Nervo Oftálmico/patologia
7.
Am J Med ; 67(4): 724-8, 1979 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-227265

RESUMO

The clinical and immunologic findings in an elderly woman with thymoma and immunoglobulin deficiency in whom cytomegalovirus (CMV) encephalitis developed are described. The patient had absent serum immunoglobulins and no circulating immunoglobulin-bearing lymphocytes. Complement receptor-bearing lymphocytes were present in the peripheral blood, and circulating T lymphocyte numbers were within normal limits. She was anergic to a battery of skin test antigens, and her lymphocytes in vitro showed a selective unresponsiveness to CMV antigen while responding normally to phytohemagglutinin and streptokinase. The course of the encephalitis was progressive with quadriplegia, aphasia and coma developing within six months of onset of symptoms. This is the fifth reported case documenting an association between CMV infection and the syndrome of thymoma with immunoglobulin deficiency, but the first report of fatal CMV encephalitis in a patient with thymoma and immunoglobulin deficiency.


Assuntos
Agamaglobulinemia/complicações , Infecções por Citomegalovirus/complicações , Encefalite/complicações , Timoma/complicações , Neoplasias do Timo/complicações , Feminino , Humanos , Pessoa de Meia-Idade
8.
Am J Med ; 61(5): 703-8, 1976 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-185900

RESUMO

Herpesvirus hominis (HVH) type 2 meningoencephalitis, confirmed by isolation of the virus from cerebrospinal fluid and brain biopsy specimens, is described in a 44 year old man following renal transplantation. An HVH type 2 genital infection developed two weeks after renal transplantation, which was followed by meningoencephalitis 10 days later. Subsequently an intracerebral hemorrhage developed with evidence of diffuse vasculitis on arteriography. In a second transplant patient a similar clinical syndrome also developed after an HVH type 2 genital infection, but viral studies were not made to confirm the etiology of the meningoencephalitis. HVH has been recognized as a cause ot mucocutaneous diseases in recipients of renal transplants, but involvement of the central nervous system has not been reported.


Assuntos
Transplante de Rim , Meningoencefalite/etiologia , Simplexvirus , Adulto , Encéfalo/microbiologia , Feminino , Humanos , Masculino , Meningoencefalite/microbiologia , Simplexvirus/isolamento & purificação , Transplante Homólogo/efeitos adversos
9.
Am J Clin Pathol ; 81(1): 110-6, 1984 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-6606977

RESUMO

Benign lymphocytic angiitis and granulomatosis (BLAG) is characterized by dense benign-appearing infiltrates of mature lymphocytes, plasma cells, and histiocytes within the pulmonary parenchyma and vasculature. The disorder typically is restricted to the lungs and has a good prognosis. The authors report a patient with BLAG and involvement of lung, kidney, and prostate. This is the first report of prominent systemic distribution in this disease. Another unique feature of this case was the presence of serum antinuclear antibodies and evidence of immune complex deposition in both lung and kidney, suggesting an underlying autoimmune disorder. An association of this entity with lymphomatoid granulomatosis (LG) has been suggested, and the prominent genitourinary disease in this patient may be indicative of a transitional stage leading to LG. An autoimmune state may be the underlying stimulus for the development of BLAG and LG.


Assuntos
Nefropatias/patologia , Pneumopatias/patologia , Granulomatose Linfomatoide/patologia , Doenças Prostáticas/patologia , Vasculite/patologia , Anticorpos Antinucleares/análise , Complexo Antígeno-Anticorpo/análise , Humanos , Nefropatias/imunologia , Pneumopatias/imunologia , Linfócitos , Granulomatose Linfomatoide/imunologia , Masculino , Pessoa de Meia-Idade , Doenças Prostáticas/imunologia , Vasculite/imunologia
10.
J Neurosurg ; 48(5): 825-8, 1978 May.
Artigo em Inglês | MEDLINE | ID: mdl-641560

RESUMO

This report describes a large, asymptomatic, subdural skeletal-type condrosarcoma originating from the meninges. Distinction between the mesenchymal and skeletal types of intracranial chondrosarcoma is important since they have different prognoses.


Assuntos
Condrossarcoma/patologia , Neoplasias Meníngeas/patologia , Adulto , Neoplasias Encefálicas/patologia , Neoplasias Encefálicas/cirurgia , Condrossarcoma/cirurgia , Feminino , Humanos , Neoplasias Meníngeas/cirurgia
11.
J Neurosurg ; 51(4): 533-8, 1979 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-383911

RESUMO

This study evaluates the histological and histochemical changes and osteogenicity of Synthos (tricalcium phosphate ceramic) implanted in the cervical intervertebral space. The cervical vertebrae from C-3 to C-6 were exposed in 20 dogs, and discectomy was performed at the third and fifth spaces. A dowel of Synthos was inserted into the third space, and a piece of fresh autologous graft from the humerus was inserted into the fifth space. The animals were divided into five groups with four animals in each. Four animals were sacrificed at each of 3, 6, 12, 18, and 22 weeks after the procedure. The C-3 to C-6 vertebrae were removed en bloc. One-half of each specimen was processed for histological examination of bone development, while the other half was processed for the two-color fluorescent labeling technique of Suzuki and Matthews for determination of osteogenesis at the time of operation and sacrifice. Various degrees of compression of the Synthos dowel were noted, with anterior and/or posterior displacement of the implant in 70% of the cases. Results indicate that the Synthos implant was biochemically and biomechanically unacceptable for the purposes of this investigation.


Assuntos
Fosfatos de Cálcio , Vértebras Cervicais/cirurgia , Disco Intervertebral/cirurgia , Fusão Vertebral/métodos , Animais , Fenômenos Biomecânicos , Transplante Ósseo , Fosfatos de Cálcio/efeitos adversos , Fosfatos de Cálcio/farmacologia , Cerâmica/efeitos adversos , Vértebras Cervicais/patologia , Cães , Disco Intervertebral/patologia , Osteogênese/efeitos dos fármacos , Complicações Pós-Operatórias/patologia , Compressão da Medula Espinal/etiologia , Compressão da Medula Espinal/patologia , Transplante Autólogo
15.
Cancer ; 38(5): 1977-82, 1976 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-991111

RESUMO

The light microscopy and ultrastructure of a malignant epithelioid schwannoma are described. Characteristic cells resembling perineural elements with various degrees of differentiation were observed. Primitive epithelioid cells contained scant ergastoplasm, and few tubules and filaments, but did have abundant free ribosomes and Golgi membranes. Also noted were junctional complexes and focal fusion of plasma membranes, basal laminae were absent. Better differentiated cells were completely limited by a well-developed basal membrane and had an abundance of intracytoplasmic filaments and multiple pinocytotic vesicles. The intercellular ground substance was composed of numerous fine collagen fibrils and amorphous, basement membrane-like, electron-dense material. A striking ultrastructural similarity of the tumor cells to those encountered in ethylnitrosourea-induced malignant schwannomas in rats was noteworthy.


Assuntos
Neoplasias Pulmonares/patologia , Neurilemoma/patologia , Adulto , Citoplasma/ultraestrutura , Feminino , Complexo de Golgi/ultraestrutura , Humanos , Neoplasias Pulmonares/ultraestrutura , Metástase Neoplásica , Neurilemoma/ultraestrutura
16.
J Biol Chem ; 276(7): 4839-44, 2001 Feb 16.
Artigo em Inglês | MEDLINE | ID: mdl-11087728

RESUMO

Auxiliary Kvbeta subunits form complexes with Kv1 family voltage-gated K(+) channels by binding to a part of the N terminus of channel polypeptide. This association influences expression and gating of these channels. Here we show that Kv4.3 proteins are associated with Kvbeta2 subunits in the brain. Expression of Kvbeta1 or Kvbeta2 subunits does not affect Kv4.3 channel gating but increases current density and protein expression. The increase in Kv4.3 protein is larger at longer times after transfection, suggesting that Kvbeta-associated channel proteins are more stable than those without the auxiliary subunits. This association between Kv4.3 and Kvbeta subunits requires the C terminus but not the N terminus of the channel polypeptide. Thus, Kvbeta subunits utilize diverse molecular interactions to stimulate the expression of Kv channels from different families.


Assuntos
Canais de Potássio de Abertura Dependente da Tensão da Membrana , Canais de Potássio/metabolismo , Animais , Encéfalo/metabolismo , Linhagem Celular , Condutividade Elétrica , Humanos , Ativação do Canal Iônico , Mutação , Canais de Potássio/genética , Subunidades Proteicas , Ratos , Canais de Potássio Shal , Transfecção
17.
Mol Pharmacol ; 45(4): 684-9, 1994 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-8183247

RESUMO

Biological studies of seven-transmembrane region G protein-coupled receptors have been restricted by available techniques for gene transfer into mammalian cells. We have created a highly efficient adenovirus-based expression vector for the thyrotropin-releasing hormone (TRH) receptor (TRH-R), AdCMVmTRHR, to circumvent difficulties encountered when transient or stable plasmid expression systems are used. We show that infection with AdCMVmTRHR results in fully functional TRH-Rs, which can be expressed in a broad range of mammalian cell types, including those resistant to conventional transient transfection. TRH-Rs can be expressed at high levels, up to 2 x 10(6) receptors/cell. Expression in several cell lines in culture reveals that rapid TRH-R desensitization by TRH and phorbol 12-myristate 13-acetate is cell type specific. The versatility of adenovirus-mediated gene transfer and expression of TRH-Rs not only facilitates in vitro studies of TRH-R biology but also provides a valuable in vivo expression vector capable of extending TRH-R studies to animal model systems.


Assuntos
Receptores do Hormônio Liberador da Tireotropina/metabolismo , Adenoviridae/genética , Animais , Regulação para Baixo/efeitos dos fármacos , Técnicas de Transferência de Genes , Vetores Genéticos , Técnicas In Vitro , Camundongos , Receptores do Hormônio Liberador da Tireotropina/genética , Acetato de Tetradecanoilforbol/farmacologia , Hormônio Liberador de Tireotropina/farmacologia
18.
J Virol ; 73(5): 3866-76, 1999 May.
Artigo em Inglês | MEDLINE | ID: mdl-10196281

RESUMO

Herpes simplex virus type 1 (HSV-1) establishes latency in sensory neurons, a state in which the viral lytic genes are silenced and only the latency locus is transcriptionally active, producing the 2. 0- and 1.5-kb latency-associated transcripts (LATs). Previous experimental evidence indicates that the LATs are stable introns, and it has been reported that LAT formation is abolished by debilitating substitution mutations in the predicted splice sites during lytic infection but not latency (J. L. Arthur et al., J. Gen. Virol. 79:107-116, 1998). We have independently studied a set of deletion mutations to explore the roles of the proposed splice sites during lytic and latent infection. HSV-1 mutant viruses missing the invariant intron-terminal 5'-G(T/C) or 3'-AG dinucleotides were analyzed for LAT formation during lytic infection in vitro, when only the 2-kb LAT is produced, and during latency in mouse trigeminal ganglia, where both LATs are expressed. Northern blot analysis of total RNAs from different productively infected cell lines showed that the lytic (2-kb) LAT was not expressed by the various splice site deletion mutants. In vivo studies using a mouse eye model of latency similarly showed that the latent (2- and 1. 5-kb) LATs were not expressed by the mutants. PCR analysis with primers flanking the LAT sequence revealed the expected splice junction for LAT excision in RNA from sensory neurons latently infected with wild-type but not mutant virus. Using a virus mutant deleted in the splicing signals flanking the 556-bp region of LAT whose absence distinguishes the 1.5- and 2-kb LATs, we observed selective elimination of 1.5-kb LAT expression in latency, supporting previous suggestions that the internal region is removed by splicing. Taken together, these results demonstrate that the 2-kb LAT is formed during both lytic and latent infection by splicing at the predicted splice sites and that an additional splicing event is involved in the latency-restricted production of the 1.5-kb LAT. We have also mapped the 3' end of the lytic 2-kb LAT and discuss our results in the context of previous models addressing the unusual stability of the LATs.


Assuntos
Herpesvirus Humano 1/genética , Splicing de RNA , RNA Viral , Latência Viral , Animais , Sítios de Ligação , Chlorocebus aethiops , Herpesvirus Humano 1/fisiologia , Humanos , Mutação , Células Tumorais Cultivadas , Células Vero
19.
Gene Ther ; 10(22): 1926-30, 2003 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-14502222

RESUMO

Ability of replication-defective adenovirus vectors to achieve efficient gene transfer in most of the mammalian cell types makes them useful vehicles for many gene transfer applications, including their use in assessing gene function. High throughput creation of recombinant adenovirus becomes a critical path to the expanding utility of adenovirus vector technology. Here, we report a process in which recombinant adenovirus vectors are isolated as single molecular clones through a convenient direct cloning and green-white selection procedure, and directly transfected into 293 cells where virus is rescued through an enzymatic reaction mediated by an intron-encoding rare endonuclease I-Sce I. This process of enzymatic rescue of circular molecular clones was at least 10-fold more efficient than that using linearized clones for transfection. This method will facilitate a high throughput creation of vectors as required for screening gene function.


Assuntos
Adenoviridae/genética , DNA Circular , Desoxirribonucleases de Sítio Específico do Tipo II/farmacologia , Engenharia Genética , Vetores Genéticos/análise , Linhagem Celular , Clonagem Molecular , Vetores Genéticos/genética , Proteínas de Fluorescência Verde , Humanos , Proteínas Luminescentes/genética , Microscopia de Fluorescência , Proteínas de Saccharomyces cerevisiae , Transfecção/métodos , Replicação Viral
20.
Gene Ther ; 1(6): 395-402, 1994 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-7584105

RESUMO

Important aspects of successful adenovirus gene transfer include the amount and persistence of gene expression, the ability to readminister virus and the localization of virus-directed gene expression to target organs. Our objective in this study was to use a single recombinant adenovirus bearing a quantifiable reporter gene [chloramphenicol acetyltransferase (CAT)] to establish the parameters which define the limits of adenovirus gene expression in a rat model. First, we determined how the route of virus administration affected the amount, duration and distribution of expression in different tissues and in rats of different developmental stages. All routes resulted in infection of all tissues tested. Surprisingly, the most efficient and widespread gene transfer was achieved by intracardiac muscle injection. The high levels of CAT protein that can be produced in a liver (< or = 1.7 mg) or a heart (< or = 196 micrograms) 5 days after infection suggest that the amount of gene product will not be a limitation in the use of adenovirus. Following peak activity at 5 days after infection, a gradual decline of CAT expression was observed in all tissues assayed; by 80 days neither CAT activity nor adenovirus DNA were detectable. In addition, adults could not be boosted by a second administration of virus, presumably due to the presence of high levels of neutralizing antibodies. The limited persistence of gene expression could be circumvented when virus was injected into neonates. Blocking T lymphocyte expansion by cyclosporine enhanced the persistence of CAT gene product over a 25-day period in heart and lung but not in liver compared with control animals.(ABSTRACT TRUNCATED AT 250 WORDS)


Assuntos
Adenoviridae/genética , Técnicas de Transferência de Genes , Animais , Cloranfenicol O-Acetiltransferase/genética , Cloranfenicol O-Acetiltransferase/metabolismo , Expressão Gênica , Genes Reporter , Terapia Genética , Sistema Imunitário/fisiologia , Ratos , Ratos Sprague-Dawley , Recombinação Genética , Linfócitos T/imunologia , Fatores de Tempo , Distribuição Tecidual
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