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2.
Clin Ter ; 167(6): e162-e166, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-28051830

RESUMO

BACKGROUND: The Gambling Disorder (GD) was recently defined as a behavioral addiction by the "The Diagnostic and Statistical Manual of Mental Disorders IV"( DSM-V) since the clinical, neurobiological and psychopathological similarities led it to be defined it as an addiction "sine substantia". The aim of this study is to formulate an "identikit" of the gambler, to evaluate a possible association between GD / emotional specific factors and the correlation between GD / substance abuse, GD / suicide. METHOD: In the study, 41 subjects were included (31 males and 10 females) and all were diagnosed with GD. A questionnaire was distributed containing 24 questions deriving from South Oaks Gambling Screen and the DSM-IVTR. RESULTS: The study showed that 51% of the respondents makes use of alcohol and / or drugs; that 73% of the patients started playing in order to relieve feelings of dysphoria and suffering consequences on work as well as family life (51%). A great deal of the respondents were indebted (39%) to the extent of needing to ask for loans from usurer (17%). Furthermore, 41% of the respondents in the sample showed that GD could be transformed into an alarming risk of suicide. DISCUSSION: The correlation between GD and drug abuse may depend on the brain function and the neural circuits that support impulsive behavior and the gratification mechanisms. Emotional experiences (stress, low level of education, divorce, poor social support) could constitute a possible risk factor that increases the GD. The committed offenses related to gambling could be explained by "loss of control". CONCLUSIONS: The results of the present study contributes to the body of knowledge regarding the size of phenomenon from a statistical and epidemiological point of view, suggesting the necessity for targeted information on the risks connected to GD in order to capture early warning signs which enables the intervention with suitable strategies.


Assuntos
Comportamento Aditivo/psicologia , Jogo de Azar/psicologia , Personalidade , Saúde Pública , Adulto , Feminino , Humanos , Comportamento Impulsivo , Masculino , Pessoa de Meia-Idade
3.
Clin Ter ; 163(4): e193-9, 2012 Jul.
Artigo em Italiano | MEDLINE | ID: mdl-23007826

RESUMO

The public health spending has now reached very significant levels, in this sense, the responsibility of the medical doctor assumes a significant importance in medical law. The aim of this paper is to analyze the profile of responsibilities of the medical doctor in the light of recent case law. The appropriateness of prescribing and risk assessment are, according to the authors, the real test on which to test the skill, prudence and diligence which are called prescribers. Guidelines can be a valuable tool for the professional help, knowing, however, limits application of the recommendations where to be reconciling with the prevailing protection of personal rights of the user.


Assuntos
Prescrições de Medicamentos/normas , Responsabilidade Legal , Itália
4.
Transplant Proc ; 43(10): 3997-9, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22172886

RESUMO

BACKGROUND: Renal cell carcinomas (RCCs) are rarely described in transplanted kidneys. Available therapeutic strategies range from allograft nephrectomy to nephron-sparing procedures such as partial nephrectomy or image-guided thermal ablation. Percutaneous radiofrequency ablation (RFA) is a minimally invasive technique which provides promising oncologic outcomes in small allograft RCCs while preserving allograft function. So far, only a few cases have been reported in the transplant setting. We describe a renal transplant RCC successfully approached by ultrasound-guided RFA. METHODS: A 42-year-old renal transplant recipient developed a small subcapsular allograft RCC at 11 years after transplantation. The decline in glomerular filtration rare prompted us to preserve as much parenchyma as possible. Ultrasound-guided RFA was performed under light sedation and local analgesia in a single session with a Starbust Talon needle. RESULTS: Postablation contrast-enhanced ultrasound displayed a 25×23 mm avascular area of complete necrosis. After 3 months gadolinium-enhanced magnetic resonance imaging confirmed the absence of viable tumor tissue and while the patient did not experience any graft function reduction (serum creatinine 2.6 mg/dL). CONCLUSIONS: Image-guided RFA represents a promising therapeutic modality for small allograft RCCs in recipients with mild graft dysfunction and/or elevated surgical risk. It is associated with low morbidity and parenchymal preservation.


Assuntos
Carcinoma de Células Renais/cirurgia , Ablação por Cateter , Neoplasias Renais/cirurgia , Transplante de Rim/efeitos adversos , Ultrassonografia de Intervenção , Adulto , Carcinoma de Células Renais/diagnóstico por imagem , Carcinoma de Células Renais/etiologia , Humanos , Neoplasias Renais/diagnóstico por imagem , Neoplasias Renais/etiologia , Masculino , Fatores de Tempo , Transplante Homólogo , Resultado do Tratamento
5.
Cancer Gene Ther ; 17(11): 814-25, 2010 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-20706287

RESUMO

Multiple endocrine neoplasia type 1 (MEN1) syndrome is characterized by the occurrence of tumors of parathyroids, neuroendocrine cells of the gastro-enteropancreatic tract and anterior pituitary. MEN1 gene encodes menin-oncosuppressor protein. Loss of heterozygosity at 11q13 is typical of MEN1 tumors. We have analyzed the MEN1 mRNA and menin expression in fibroblasts from normal skin biopsies and from MEN1 patients (two with a frameshift 738del4 (exon 3) mutation, introducing a premature stop codon, and an individual with an R460X (exon 10) nonsense mutation). The expression of full-length menin protein did not differ between MEN1 and normal fibroblasts. Wild-type alleles mRNAs were expressed in MEN1 patients, whereas mutant alleles were partially degraded by nonsense-mediated mRNA decay pathway, suggesting a mechanism of compensation for allelic loss by the up-regulation of wild-type menin expression at a post-transcriptional level. Small-interfering RNA silencing of the wild-type mRNA allele abolished menin compensation, whereas the ribozyme silencing of the MEN1-mutated mRNA allele resulted in strongly enhanced wild-type menin expression. Gel-retardation analysis showed that in vitro-specific RNA-protein complexes bound to MEN1 mRNA. These findings contribute to the understanding of tumorigenesis in MEN1, offering the basis for the development of RNA-based therapies in MEN1 gene mutation carriers.


Assuntos
Fibroblastos/metabolismo , Neoplasia Endócrina Múltipla Tipo 1/metabolismo , Proteínas Proto-Oncogênicas/genética , RNA Catalítico/metabolismo , RNA Interferente Pequeno/metabolismo , Adulto , Caspase 8/metabolismo , Éxons , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Perda de Heterozigosidade , Masculino , Pessoa de Meia-Idade , Neoplasia Endócrina Múltipla Tipo 1/genética , Mutação , Proteínas Proto-Oncogênicas/metabolismo , Interferência de RNA , RNA Mensageiro/metabolismo , Células Tumorais Cultivadas , Regulação para Cima
6.
Bioelectrochemistry ; 67(2): 135-41, 2005 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-16027048

RESUMO

Two biosensors have been constructed using an RNA aptamer as biorecognition element. The aptamer, specific for HIV-1 Tat protein, has been immobilised on the gold surface of piezoelectric quartz crystals or surface plasmon resonance (SPR) chips to develop a quartz crystal microbalance (QCM)-based and an SPR-based biosensor, respectively. Both the biosensors were modified with the same immobilisation chemistry based on the binding of a biotinylated aptamer on a layer of streptavidin. The binding between the immobilised aptamer and its specific protein has been evaluated with the two biosensors in terms of sensitivity, reproducibility and selectivity. A protein very similar to Tat, Rev protein, has been used as negative control. The two biosensors both were very reproducible in the immobilisation and the binding steps. The selectivity was high in both cases.


Assuntos
Técnicas Biossensoriais , Produtos do Gene tat/análise , HIV-1 , Quartzo/química , RNA/química , Ressonância de Plasmônio de Superfície , Produtos do Gene tat/química , Sensibilidade e Especificidade , Produtos do Gene tat do Vírus da Imunodeficiência Humana
7.
Biosens Bioelectron ; 20(6): 1149-56, 2004 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-15556361

RESUMO

The in vitro selection of combinatorial libraries of RNA/DNA, has allowed the identification of specific nucleic acids (aptamers) which bind to a wide range of target molecules with high affinity and specificity. In this work, an RNA aptamer, specific for the protein trans-activator of transcription (Tat) of HIV-1, has been used as bio-recognition element to develop a biosensor (aptasensor). The biosensor was optimised using piezoelectric quartz-crystals as transducers and the aptamer was immobilised on the gold electrode of the crystal. The immobilisation procedure was based on the interaction between the biotinylated aptamer and streptavidin previously deposited on the electrode. The main analytical characteristics of the biosensor, such as sensitivity, selectivity and reproducibility, have been studied in details. An optimised regeneration procedure allowed the multiple use of the aptamer-coated crystal. The aptasensor has been compared with the corresponding immunosensor, based on the specific monoclonal anti-Tat antibody. The antibody was immobilised on a layer of carboxylated dextran previously deposited on the gold electrode. The results demonstrated that the use of a biosensor with a specific aptamer as bio-recognition element could be an interesting approach in the detection of proteins, which has been here examined considering a model system.


Assuntos
Técnicas Biossensoriais/instrumentação , Materiais Revestidos Biocompatíveis/química , Eletroquímica/instrumentação , Produtos do Gene tat/análise , Produtos do Gene tat/química , HIV-1/metabolismo , RNA/química , Técnicas Biossensoriais/métodos , Eletroquímica/métodos , Desenho de Equipamento , Análise de Falha de Equipamento , Proteínas de Ligação a RNA/análise , Proteínas de Ligação a RNA/química , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Produtos do Gene tat do Vírus da Imunodeficiência Humana
8.
Nucleic Acids Res ; 28(21): 4059-67, 2000 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-11058100

RESUMO

A library of three synthetic ribozymes with randomized arms, targeting NUX, GUX and NXG triplets, respectively, were used to identify ribozyme-accessible sites on the HIV-1 LTR transcript comprising positions -533 to 386. Three cleavable sites were identified at positions 109, 115 and 161. Ribozymes were designed against these sites, either unmodified or with 2'-modifications and phosphorothioate groups, and their cleavage activities of the transcript were determined. Their biological activities were assessed in cell culture, using a HIV-1 model assay system where the LTR is a promoter for the expression of the reporter gene luciferase in a transient expression system. Intracellular efficiency of the ribozymes were determined by cotransfection of ribozyme and plasmid DNA, expressing the target RNA. Modified ribozymes, directed against positions 115 and 161, lowered the level of LTR mRNA in the cell resulting in inhibition of expression of the LTR-driven reporter gene luciferase of 87 and 61%, respectively. In the presence of Tat the inhibitions were 43 and 25%. The inactive variants of these ribozymes exhibited a similar inhibitory effect. RNase protection revealed a reduction of RNA which was somewhat stronger for the active than the inactive ribozymes, particularly for ribozyme 115. Unmodified ribozymes showed no inhibition in the cell. The third ribozyme, targeting a GUG-triplet at position 109, possessed only low cleavage activity in vitro and no inhibitory effect in cell culture.


Assuntos
Regulação para Baixo , Regulação Viral da Expressão Gênica , Repetição Terminal Longa de HIV/genética , HIV-1/genética , RNA Catalítico/genética , RNA Catalítico/metabolismo , Sequência de Bases , Sítios de Ligação , Códon/genética , Biblioteca Gênica , Genes Reporter/genética , Engenharia Genética , Células HeLa , Humanos , Cinética , Ensaios de Proteção de Nucleases , Conformação de Ácido Nucleico , Oligorribonucleotídeos/síntese química , Oligorribonucleotídeos/química , Oligorribonucleotídeos/genética , Oligorribonucleotídeos/metabolismo , Estabilidade de RNA , RNA Catalítico/síntese química , RNA Catalítico/química , RNA Mensageiro/análise , RNA Mensageiro/genética , RNA Viral/química , RNA Viral/genética , RNA Viral/metabolismo , Especificidade por Substrato , Transcrição Gênica/genética , Transfecção
9.
Trends Biotechnol ; 16(10): 434-8, 1998 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-9807841

RESUMO

Ribozymes are being increasingly used for the sequence-specific inhibition of gene expression by the cleavage of mRNAs encoding proteins of interest. However, particular attention must be paid to the following points: the identification of regions on the mRNA accessible to the ribozyme; the delivery of ribozymes to cells by either exogenous or endogenous delivery; colocalization of the ribozyme with the target RNA in the cell; and differentiation between closely related sequences. This field is advancing rapidly, and results obtained with transgenic animals demonstrate the power of this strategy for the inhibition of gene expression.


Assuntos
Regulação da Expressão Gênica/efeitos dos fármacos , Vetores Genéticos/genética , RNA Catalítico/química , RNA Catalítico/farmacologia , Animais , Portadores de Fármacos , Sistemas de Liberação de Medicamentos , Vetores Genéticos/farmacologia , Humanos , RNA/efeitos dos fármacos , RNA Catalítico/genética
10.
Proc Natl Acad Sci U S A ; 94(18): 9711-6, 1997 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-9275189

RESUMO

Strand-specific transcripts of a satellite DNA of the newts, Notophthalmus and Triturus, are present in cells in monomeric and multimeric sizes. These transcripts undergo self-catalyzed, site-specific cleavage in vitro: the reaction requires Mg2+ and is mediated by a "hammerhead" domain. Transcription of the newt ribozyme appears to be performed by RNA polymerase II under the control of a proximal sequence element and a distal sequence element. In vitro, the newt ribozyme can cleave in trans an RNA substrate, suggesting that in vivo it might be involved in RNA processing events, perhaps as a riboprotein complex. Here we show that the newt ribozyme is in fact present as a riboprotein particle of about 12 S in the oocytes of Triturus. In addition, reconstitution experiments and gel-shift analyses show that a complex is assembled in vitro on the monomeric ribozyme molecules. UV cross-linking studies identify a few polypeptide species, ranging from 31 to 65 kDa, associated to the newt ribozyme with different affinities. Finally, we find that an appropriate oligoribonucleotide substrate is specifically cleaved by the riboproteic activity in S-100 ovary extracts.


Assuntos
RNA Catalítico/química , Ribonucleoproteínas/química , Animais , DNA/genética , DNA/metabolismo , Notophthalmus , RNA Catalítico/genética , RNA Catalítico/metabolismo , Ribonucleoproteínas/genética , Ribonucleoproteínas/metabolismo , Transcrição Gênica , Triturus
11.
Eur J Biochem ; 247(1): 396-401, 1997 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-9249052

RESUMO

To analyse the trans-cleavage activity of the hammerhead ribozyme occurring in the ovary of the newt (Notophthalmus, Triturus) in more detail, six synthetic ribozymes representing natural and modified hammerhead sequences were tested with both short oligoribonucleotides and long transcripts as substrates. The same analysis was also performed with the monomer (330 nucleotides) newt ribozyme and variants thereof. None of the ribozymes comprising the newt natural sequence showed activity under multiple turnover conditions, regardless of sequence changes in stem and loop II. With excess of ribozyme, the same ribozymes cleaved only to a limited extent a short substrate and extremely poorly a target site embedded within a long transcript. The addition of whole ovary cell extract had little influence on cleavage activity of short substrates. However, sequence changes in stems I and III to target different sequences considerably improved cleavage ability of the ribozymes under all conditions used. An RNA secondary-structure folding program showed that ribozymes with the natural newt sequence did not fold in a hammerhead structure whereas those with the changes in stem I and III did. These results suggest that the sequence of the stems I and III impairs the assembly of the newt ribozyme into a bimolecular hammerhead complex in vitro and that proteins present in the ovaries do not facilitate activity.


Assuntos
RNA Catalítico/metabolismo , Salamandridae/metabolismo , Animais , Sequência de Bases , Dados de Sequência Molecular , Dobramento de Proteína , RNA Catalítico/química
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