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1.
World J Surg ; 38(11): 2940-5, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24889413

RESUMO

BACKGROUND: Cysts in contact with the inferior vena cava (IVC) represent a challenge for hepato-pancreatico-biliary surgeons. Although the literature on the topic is scarce, the most widely accepted approach is conservative surgery. Partial cyst resection is recommended, because radical resection is considered a high-risk procedure. STUDY DESIGN: This was a retrospective study over the period January 2007-December 2012. We operated on 103 patients with liver hydatidosis. A total of 32 patients (31 %) had a liver cyst in contact with the IVC. We proposed a cyst classification based on location of the cyst and length of contact and degrees of involvement of the IVC. RESULTS: Median size of the contacting cyst measured by computed tomography (CT) was 12 cm. On CT, median length of contact with the IVC was 37 mm. The median degree of involvement was 90°. Radical surgery was performed in 20 patients (62.5 %). No IVC resection was done. Morbidity rate was 28 %, and mortality was 3 %. In follow-up (median 27 months), no relapses or problems related to IVC flow were detected. Postoperative stay and transfusion rate were higher in the conservative surgery group, but these patients presented fewer complications. There was no relationship between circumferential grades and length of contact with the IVC and the type of surgery performed. CONCLUSIONS: Liver hydatid cysts in contact with the IVC are large cysts usually located in the right liver. They do not normally cause clinical symptoms related to IVC contact. Radical surgery is feasible, and was performed in 60 % of our series, but it is technically demanding. We propose a classification of cysts in contact with the IVC.


Assuntos
Equinococose Hepática/cirurgia , Hepatectomia/métodos , Veia Cava Inferior/cirurgia , Adulto , Idoso , Idoso de 80 Anos ou mais , Equinococose Hepática/diagnóstico por imagem , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/cirurgia , Estudos Retrospectivos , Tomografia Computadorizada por Raios X , Veia Cava Inferior/diagnóstico por imagem , Adulto Jovem
2.
Clin Transl Oncol ; 19(7): 915-920, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28144828

RESUMO

PURPOSE: To describe the frequency of head and/or pancreas uncinate process uptake of 99mTc-HYNIC-TOC, to study its nature, and analyze its diagnostic value. MATERIALS AND METHODS: Retrospective evaluation of 47 consecutive 99mTc-HYNIC-TOC examinations was conducted. Head and/or pancreas uncinate process uptake was considered to be physiological in patients with normal CT at the same episode and in follow-up. It was analyzed if age or diabetes mellitus was justifying the existence or not of uptake. RESULTS: 32.5% patients showed uptake; 73% of them were mild. 84.6% patients with uptake have no pathology and 4% had neuroendocrine pancreatic disease at CT. Neither the age nor the diabetes mellitus established differences in patients without lesion. CONCLUSIONS: Near one-third of patients show physiological uptake by head and/or pancreas uncinate process at 99mTc-HYNIC-TOC scintigraphy. It seems that neither the diabetes nor the ages are factors that determine this physiological uptake.


Assuntos
Tumores Neuroendócrinos/diagnóstico por imagem , Tumores Neuroendócrinos/metabolismo , Octreotida/análogos & derivados , Compostos de Organotecnécio/metabolismo , Pâncreas/diagnóstico por imagem , Pâncreas/metabolismo , Cintilografia/métodos , Receptores de Somatostatina/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Tumores Neuroendócrinos/patologia , Octreotida/metabolismo , Pâncreas/patologia , Prognóstico , Estudos Retrospectivos , Tomografia Computadorizada por Raios X
3.
Rev Esp Quimioter ; 12(4): 317-24, 1999 Dec.
Artigo em Espanhol | MEDLINE | ID: mdl-10855010

RESUMO

Genotypes that confer drug resistance to reverse transcriptase inhibitors and protease inhibitors were evaluated in HIV-1 proviral DNA obtained from peripheral blood mononuclear cell samples. Fifty-three HIV-1-infected patients were studied, 19 of whom had not received antiretroviral treatment. In the other 34 patients, 9 had been treated with combinations of two reverse transcriptase inhibitors (AZT, ddI, d4T, 3TC) and 25 had been treated with triple antiretroviral therapy including a protease inhibitor (nelfinavir, indinavir, saquinavir, ritonavir). To determine the presence of mutations involved in the development of resistance to reverse transcriptase inhibitors a hybridization Microtiter assay was carried out. Mutations were detected in treated patients as well as in those without previous antiretroviral treatment, with the most frequent mutations being those that confer resistance to AZT, followed by those that develop cross-resistance to ddI/ddC and 3TC, which are the most commonly used drugs to date. No mutations were detected to any nucleoside analog in only 13 cases. To analyze the presence of mutations in the protease gene a dot-blot hybridization was carried out which included the mutations in codons 36, 82 and 90. Mutation 82 was detected in one case. Therefore, with the aim of determining the pattern of genotypic mutations in patients infected with HIV-1 and in order to make the best therapeutic choice, it would be recommended to consider carrying out genotypic resistance assays in clinical practice.


Assuntos
Fármacos Anti-HIV/uso terapêutico , Infecções por HIV/tratamento farmacológico , HIV-1/efeitos dos fármacos , HIV-1/genética , Nucleosídeos/uso terapêutico , Inibidores de Proteases/uso terapêutico , Fármacos Anti-HIV/farmacologia , Resistência Microbiana a Medicamentos , Humanos , Mutação , Nucleosídeos/farmacologia , Inibidores de Proteases/farmacologia
4.
An Med Interna ; 8(8): 372-6, 1991 Aug.
Artigo em Espanhol | MEDLINE | ID: mdl-1768745

RESUMO

We have investigated, by "in situ" hibridisation, the presence of hepatitis B virus (HBV) DNA in peripheral blood mononuclear cells (PBMC) from 45 patients with acute and chronic hepatic disorders directly related with HBV or with some seric HBV marker. Results has been related with serological markers and the different types of hepatopaties. The HBV-DNA was detected in PBMC more frequently in patients with hepatic alterations more prolongated (chronic active hepatitis, chronic persistent hepatitis and cirrhosis) than in acute hepatitis patients. It was not detected in any asymptomatic patient with HBV serological markers. As regards HBV serological markers, HBV-DNA was detected in PBMC in 8/11 HBsAg positive patients and in 11/34 HBsAg negative patients: 3 antiHBc positive, 5 antiHBc and antiHBs positive and 3 without conventional seric markers. The detection of HBV-DNA in antiHBc and/or antiHBs positive subjects means the virus may persist after recovery of infection and suggests PMBC could serve as additional reservoirs for reinfection of hepatocytes leading to a reactivation of the liver disease. Our results suggest that HBV infection of PBMC is a frequent event during HBV infection and can have important consequences fundamentally with respect to pathogenic mechanisms of HBV induced liver disease and to the transmission of the virus.


Assuntos
DNA Viral/análise , Vírus da Hepatite B/isolamento & purificação , Hepatite B/microbiologia , Leucócitos Mononucleares/microbiologia , Hepatopatias/complicações , Doença Aguda , Adulto , Criança , Hepatite B/complicações , Anticorpos Anti-Hepatite B/análise , Antígenos da Hepatite B/análise , Vírus da Hepatite B/genética , Vírus da Hepatite B/imunologia , Hepatite Crônica/complicações , Humanos , Leucócitos Mononucleares/química , Hepatopatias/sangue , Hibridização de Ácido Nucleico
5.
An Med Interna ; 7(9): 456-8, 1990 Sep.
Artigo em Espanhol | MEDLINE | ID: mdl-2129411

RESUMO

We found the presence of hepatitis B virus in 17 cases of non-A-non-B hepatitis using the DNA detection technique in serum of patients with a type of chronic hepatopathy. This finding supports the needs to determine this seric marker in all patients afflicted with chronic hepatopathy before the diagnosis of hepatitis B is excluded.


Assuntos
DNA Viral/sangue , Vírus da Hepatite B/genética , Hepatite C/diagnóstico , Adulto , Biomarcadores/sangue , Doença Crônica , Sondas de DNA , Feminino , Humanos , Immunoblotting , Masculino , Pessoa de Meia-Idade , Hibridização de Ácido Nucleico
6.
An Med Interna ; 7(10): 505-12, 1990 Oct.
Artigo em Espanhol | MEDLINE | ID: mdl-2104095

RESUMO

We study retrospectively the viral replication state (HBV) of 50 patients with chronic hepatic alterations. The seric DNA-HBV and/or intrahepatic (molecular hybridization), the intrahepatic distribution of HBV antigens (specific monoclonal antibodies labelled with immunoperoxidase), conventional seric HBV markers (commercial enzymoimmunoessay) and the different histopathologic features. We found a correlation between DNA-HBV "in situ" and HBcAg intrahepatic and the seric DNA-HBV production. 81% of the patients with HBsAg (+) had intrahepatic HBcAg and 85% (11/13) of them showed the antigen in their cytoplasms. Patients with HBcAg also had seric and liver DNA-HBV (+). The lack of seric HBsAg did not mean that non-active replication of HBV did not exist because 20% of the patients with HBsAg (-) showed seric and "in situ" DNA-HBV and cytoplasmic HBcAg. The detection of DNA-HBV in endothelial cells and vascular elements in hepatic tissue show that the rate of the HBV host cells is greater.


Assuntos
Vírus da Hepatite B/fisiologia , Hepatite/microbiologia , Replicação Viral , Adolescente , Adulto , Idoso , Biomarcadores , Criança , Pré-Escolar , Doença Crônica , DNA Viral/análise , Feminino , Hepatite/sangue , Anticorpos Anti-Hepatite B/análise , Antígenos da Hepatite B/análise , Humanos , Lactente , Fígado/química , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos
7.
An Sist Sanit Navar ; 23(1): 35-40, 2000.
Artigo em Espanhol | MEDLINE | ID: mdl-12886319

RESUMO

AIM: The aim of this study is to determine the nature of venous flow in the lower extremities, as well as to correlate the velocity of the flow with average vein pressure in the right atrium. MATERIAL AND METHODS: Over a period of one year 236 pulsated Doppler echographs were made of patients, bearers of a venous catheter located in the right atrium. The patients were in a supine lying position and breathing gently. The lines of the Doppler wave were used to analyse the frequency of the wave, the components of its velocity, the relationships of velocity and the existence of pulsatile flow. These parameters were compared with the pressure in the right atrium. RESULTS: The study showed a cardiac and respiratory periodicity of the venous wave. A statistically significant correlation (p<0.0001) was found between average venous pressure in the right atrium and the following variables: the systolic atrium wave (a), the diastolic atrium wave (d), the relation of pulsatility (RP=Velocity min/Velocity max) and the index of pulsatility (IP=[Velocity max-Velocity min]/Velocity average). CONCLUSION: There is an inverse and significant relationship between pulsatile flow and atrium pressure. Nonetheless, although a relation exists between the different components of the venous wave, an elevation in central vein pressure cannot be predicted.

8.
JBR-BTR ; 94(5): 287-9, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22191294

RESUMO

Aveolar soft part sarcoma is a rare tumor responsible for about 1% of all soft tissue sarcomas, affecting mostly adolescents and young adults. ASPS has curious patterns of metastatic spread, with seldom lymph node involvement. Lung, bone and brain are the most common metastatic places. Small bowel metastasis are infrequent, having found reported only one case of duodenal metastasis with polypous appearance. We describe a case of duodenal metastasis presenting as abdominal mass five years after initial diagnosis of alveolar soft part sarcoma.


Assuntos
Neoplasias Duodenais/secundário , Sarcoma Alveolar de Partes Moles/patologia , Neoplasias de Tecidos Moles/patologia , Adulto , Neoplasias Duodenais/cirurgia , Feminino , Humanos , Perna (Membro)
9.
Enferm Infecc Microbiol Clin ; 11(5): 250-4, 1993 May.
Artigo em Espanhol | MEDLINE | ID: mdl-8324021

RESUMO

BACKGROUND: The aim of this study was to develop molecular biology techniques as hybridization with oligonucleotide probes to characterize TEM-1 and TEM-2 beta-lactamases in strains of Salmonella spp. METHODS: Twenty seven strains of Salmonella spp. were selected. Twenty six were resistant to ampicillin due to the production of beta-lactamases enzymes of pl of 5.4 and/or 5.6 corresponding to TEM-type. Initially, they were submitted to colony hybridization with a 420 bp. TEM probe obtained from plasmid pBR322. The strains with positive signal were selected to perform colony hybridization and Southern blot with oligonucleotide probes for TEM-1 (Gln 37) and TEM-2 (Lys 37). Finally, polymerase chain reaction technique (PCR) was developed to obtain DNA. RESULTS: Only 17 out of the 26 beta-lactamase producing strains gave positive signals with the TEM intragenic probe. Experiments with the oligonucleotide probes in colony hybridization did not allow us discriminate positive from negative signals. Southern blot of DNA obtained from alkaline lysates did not work as we could not obtain any signals in the filters. To resolve these problems and to obtain enough DNA to perform Southern blot we developed PCR. This way it was able discriminate the bla-TEM-1 from the bla-TEM-2 genes. CONCLUSIONS: PCR technique plus oligonucleotide probes are a good alternative for the specific characterization of TEM-1 and TEM-2 beta-lactamases in Salmonella spp.


Assuntos
Sondas de DNA , Salmonella/enzimologia , beta-Lactamases/isolamento & purificação , Resistência a Ampicilina , Reação em Cadeia da Polimerase , Salmonella/efeitos dos fármacos , Salmonella/genética
10.
Enferm Infecc Microbiol Clin ; 11(4): 187-9, 1993 Apr.
Artigo em Espanhol | MEDLINE | ID: mdl-8512969

RESUMO

BACKGROUND: It was studied the production of HBV from lymphocytes from peripheral blood induced with a mitogen agent as phytohemagglutinin (PHA), in patients infected by HIV. METHODS: The methodology developed included the culture of peripheral blood mononuclear cells and its induction to proliferate with PHA, in order to study the viral activity in the supernatants of the cultures by detection of HIV-Ag (ELISA), HBsAg (ELISA) and DNA-HBV (Dot-blot). RESULTS: Production of HBV in 23 out of the 42 patients included in this study, corresponding to 21 anti-HIV positive and 2 anti-HIV negative. CONCLUSIONS: Induction to proliferate lymphocytes with a mitogen agent as PHA may represent an alternative way of culture to study interactions between HIV and HBV.


Assuntos
Vírus da Hepatite B/crescimento & desenvolvimento , Ativação Linfocitária/efeitos dos fármacos , Linfócitos/microbiologia , Fito-Hemaglutininas/farmacologia , Ativação Viral/efeitos dos fármacos , Células Cultivadas , DNA Viral/análise , Infecções por HIV/complicações , Hepatite B/sangue , Hepatite B/complicações , Hepatite B/microbiologia , Antígenos de Superfície da Hepatite B/análise , Vírus da Hepatite B/isolamento & purificação , Vírus da Hepatite B/fisiologia , Humanos , Linfócitos/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos
11.
Enferm Infecc Microbiol Clin ; 11(2): 80-3, 1993 Feb.
Artigo em Espanhol | MEDLINE | ID: mdl-8481440

RESUMO

BACKGROUND: The purpose of this study was to develop a satisfactory technique to detect punctual mutations in blaTEM genes. METHODS: The strains [E. coli HB 101 pBR322 (TEM-1), E. coli J62 RP4 (TEM-2)] were submitted to PCR with primers PL1 and PL2 which amplify the genetic region susceptible of punctual mutations. Then, we developed Southern blot and hybridization with oligonucleotide probes (GIn 37, Lys 37 y Thr 261), corresponding to first and last mutations. RESULTS: A region of 841 bp was amplified using the primers previously described. Hybridization experiments with the Thr 261 probe gave positive signal with both strains (both carry the mutation); GIn 37 only hybridized with TEM-1 and Lys 37 only with TEM-2. CONCLUSIONS: The use of primers which amplify all the region susceptible of mutations in blaTEM and oligonucleotide probes allows the specific detection of point modifications in the original genes by the use of digoxigenin-labeled oligonucleotide probes.


Assuntos
Proteínas de Bactérias/genética , Análise Mutacional de DNA , Escherichia coli/genética , Genes Bacterianos , Reação em Cadeia da Polimerase , beta-Lactamases/genética , Adamantano/análogos & derivados , Sequência de Bases , DNA Bacteriano/genética , Digoxigenina , Escherichia coli/enzimologia , Medições Luminescentes , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/métodos
12.
Enferm Infecc Microbiol Clin ; 10(8): 445-50, 1992 Oct.
Artigo em Espanhol | MEDLINE | ID: mdl-1489771

RESUMO

HBV infection has been investigated in 47 anti-human HIV positive patients in relation to a similar group of 33 anti-HIV negative patients. Serological HBV markers were found in 87% of anti-HIV positive patients. The difference in markers of viral replication (HBeAg, HBV-DNA) was not statistically significant between the two groups. It has been suggested that HBV infection of peripheral blood mononuclear cells could be a cofactor implicated in the development of immunodeficiency due to HIV. For this reason we have investigated the presence of HBV-DNA in peripheral blood mononuclear cells by in situ hybridization. Although its detection was more frequent in anti-HIV positive patients than in anti-HIV negative ones (p < 0.05), it was not related to clinical state of immunodeficiency. With regard to serological HBV markers, HBV-DNA was detected in peripheral blood mononuclear cells from antiHBc w/o antiHBs patients. This fact means the virus may persist in this cells after recovery and suggest they could serve as additional reservoirs of HBV. These cells, that contain the HBV genome, could be implicated in the perpetuation, reactivation of the infection and in its transmission.


Assuntos
DNA Viral/sangue , Infecções por HIV/complicações , Anticorpos Anti-Hepatite B/sangue , Antígenos da Hepatite B/sangue , Vírus da Hepatite B/isolamento & purificação , Hepatite B/diagnóstico , Leucócitos Mononucleares/microbiologia , Adulto , Feminino , Anticorpos Anti-HIV/sangue , Hepatite B/complicações , Hepatite B/epidemiologia , Hepatite B/transmissão , Vírus da Hepatite B/imunologia , Vírus da Hepatite B/fisiologia , Humanos , Masculino , Prevalência , Ativação Viral , Replicação Viral
13.
Enferm Infecc Microbiol Clin ; 10(8): 470-3, 1992 Oct.
Artigo em Espanhol | MEDLINE | ID: mdl-1489775

RESUMO

BACKGROUND: The presence of hepatitis delta virus was investigated in liver biopsies by in situ cyto-hybridization with a probe labelled with digoxigenin. METHODS: The techniques developed included extraction of plasmid DNA by lysis by alkali, electroelution, electrophoresis in agarose gel and digoxigenin labelling, and the application in liver tissue. RESULTS: Viral RNA was detected in 6 of the 10 patients, and the reactivity was exclusively restricted to the nucleolus of the hepatocytes. CONCLUSION: This method reveals as a sensitive and quick diagnostic procedure, which allows to study the intensity of the infection as well as the serological state of the patient.


Assuntos
Sondas de DNA , Digoxigenina , Hepatite D/microbiologia , Vírus Delta da Hepatite/isolamento & purificação , Hibridização In Situ , Doença Aguda , Colorimetria , DNA/genética , DNA Recombinante , Vírus Defeituosos/isolamento & purificação , Anticorpos Anti-Hepatite/sangue , Hepatite B/complicações , Hepatite D/complicações , Hepatite D/diagnóstico , Vírus Delta da Hepatite/imunologia , Hepatite Crônica/microbiologia , Humanos , Cirrose Hepática/microbiologia , Nitroazul de Tetrazólio , Plasmídeos , RNA Viral/análise
14.
Eur Radiol ; 10(12): 1920-5, 2000.
Artigo em Inglês | MEDLINE | ID: mdl-11305571

RESUMO

The aim of this study was to evaluate sonographic signs described for carpal tunnel syndrome (CTS). Sixty-four wrists from 40 patients with CTS confirmed by electromyography, and 42 wrists from 24 healthy individuals, were examined using sonography. Cross-sectional area, flattening ratio in proximal, middle and distal segments of the carpal median nerve and bowing of the flexor retinaculum were measured. The accuracies of the sonographic diagnostic criteria for CTS were assessed using receiver-operating-characteristic (ROC) analytical techniques. A significant swelling of the median nerve was observed at the proximal (p < 0.001), middle (p < 0.0001) and distal (p< 0.0001) segments and a significant bowing of the flexor retinaculum in CTS patients with respect to healthy subjects. No significant differences were found in the mean value of flattening ratio between the groups. The sensitivity, specificity, positive predictive value, and the negative predictive value were 73.4, 57.1, 72.3 and 58.5%, respectively, in the proximal and middle segments; 75, 57.1, 72.7 and 60% in the distal segment for areas greater than 11 mm2: and 81.3, 64.3, 77.6 and 69.2% for the bowing of the flexor retinaculum greater than 2.5 mm. All sonographic criteria were found in 34 CTS patients (53.1%) and none in 3 patients. Sonography may be useful in the diagnosis of CTS. The most reliable sign was increased bowing of the flexor retinaculum and cross-sectional area of median nerve with specificity close to 60%.


Assuntos
Síndrome do Túnel Carpal/diagnóstico por imagem , Adulto , Idoso , Feminino , Humanos , Masculino , Nervo Mediano/diagnóstico por imagem , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Curva ROC , Sensibilidade e Especificidade , Ultrassonografia , Articulação do Punho/diagnóstico por imagem
15.
AJR Am J Roentgenol ; 170(1): 183-5, 1998 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-9423628

RESUMO

OBJECTIVE: Testicular cysts of the tunica albuginea are an uncommon phenomenon, important because of their possible confusion with testicular tumors. Accurate diagnosis may avoid aggressive and irreversible treatment such as testicular ablation. The purpose of this study is to report our experience in such diagnosis through sonographic detection of albuginea cysts. CONCLUSION: Sonography is the technique of choice in the diagnosis of any testicular tumor formation, allowing the differentiation of cysts from neoplasia and avoiding unnecessary intervention in patients with cysts of the tunica albuginea.


Assuntos
Cistos/diagnóstico por imagem , Doenças Testiculares/diagnóstico por imagem , Adulto , Diagnóstico Diferencial , Humanos , Masculino , Neoplasias Testiculares/diagnóstico por imagem , Ultrassonografia
16.
Enferm Infecc Microbiol Clin ; 8(1): 11-4, 1990 Jan.
Artigo em Espanhol | MEDLINE | ID: mdl-1710507

RESUMO

The diagnosis of human immunodeficiency virus infection is sometimes difficult or nonspecific, both in early and late stages, as the patients may be seronegative at the time of testing. Although serologic testing usually suffices to identify infected individuals and to follow up the course of the infection, in some cases direct detection of the virus is required. The culture of the mononuclear cells of the patient permits, after stimulation with mitogens and interleukin-2, the expression of viral antigens even in asymptomatic patients with latent or apparently nonproductive infection. In this way we have recovered the virus in four patients without serological evidence of infection. The possibility that human immunodeficiency virus infection can be undetectable with the usual diagnostic techniques, at least in a small proportion of patients, supports the need to use other methods such as direct viral culture to permit the identification of a greater number of infected individuals and the adoption of the appropriate prophylactic or therapeutic measures.


Assuntos
Sorodiagnóstico da AIDS , Infecções por HIV/diagnóstico , Soropositividade para HIV/diagnóstico , HIV/isolamento & purificação , Adulto , Reações Falso-Negativas , Antígenos HIV/análise , Infecções por HIV/enzimologia , Infecções por HIV/microbiologia , Soropositividade para HIV/enzimologia , Humanos , DNA Polimerase Dirigida por RNA/metabolismo , Cultura de Vírus
17.
Enferm Infecc Microbiol Clin ; 10(4): 220-3, 1992 Apr.
Artigo em Espanhol | MEDLINE | ID: mdl-1606226

RESUMO

BACKGROUND: Production of DNA probes is time-consuming and inefficient. We have developed a method for the obtention of digoxigenin-labeled probes to detect TEM-type betalactamases using the polymerase chain reaction. The amplification product was a 516 bp fragment internal to bla-TEM-1 from pBR 322. METHODS: The techniques developed included extraction of plasmid DNA by lisis by alkali, electroelution, electrophoresis in agarose gels, polymerase chain reaction and hydridization with a DNA probe digoxigenin labeled. RESULTS: We obtained by polymerase chain reaction 1500 ng of probe using 1 ng of target DNA. Developing classic methods the amount of probe was 75 ng from 1 microgram of target DNA. The time to obtain the probe was 3 hours, instead of a week with other methods. CONCLUSIONS: We conclude that polymerase chain reaction is a good alternative to classic methods to obtain digoxigenin-labeled DNA probes.


Assuntos
Sondas de DNA/isolamento & purificação , Reação em Cadeia da Polimerase , beta-Lactamases/análise , Sequência de Bases , Digoxigenina , Dados de Sequência Molecular
18.
Eur J Clin Microbiol Infect Dis ; 14(7): 591-6, 1995 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-7588843

RESUMO

To investigate the association of human immunodeficiency virus (HIV) with various DNA viruses, including hepatitis B virus (HBV), cytomegalovirus (CMV) and Epstein-Barr virus, (EBV), simultaneous detection of HIV p24 antigen, HBV surface antigen and DNA, CMV-DNA and EBV-DNA expression was performed in phytohemagglutinin-stimulated peripheral blood mononuclear (PBMC) culture supernatants obtained from 54 individuals at risk for HIV infection. HIV expression in PBMC culture supernatants never occurred alone; expression of other viruses was always detected in the 24 samples expressing HIV antigen in vitro. Furthermore, in 16 patients expression of other viruses was detected without HIV expression, and in 14 patients none of the tested viruses were detected. These results indicate a strong association between the presence of HIV antibody and expression of DNA viruses in vitro (p = 0.0001). The coexpression of these viruses could be related to the evolution of HIV infection and AIDS.


Assuntos
Citomegalovirus/isolamento & purificação , Infecções por HIV/virologia , Vírus da Hepatite B/isolamento & purificação , Herpesvirus Humano 4/isolamento & purificação , Células Cultivadas , Citomegalovirus/genética , Citomegalovirus/imunologia , DNA Viral/análise , Proteína do Núcleo p24 do HIV/análise , Vírus da Hepatite B/genética , Vírus da Hepatite B/imunologia , Herpesvirus Humano 4/genética , Herpesvirus Humano 4/imunologia , Humanos , Leucócitos Mononucleares/virologia
19.
Enferm Infecc Microbiol Clin ; 10(10): 597-601, 1992 Dec.
Artigo em Espanhol | MEDLINE | ID: mdl-1292600

RESUMO

BACKGROUND: The purpose of this work was to study the molecular basis of beta-lactamase production in ampicillin-resistant strains of Salmonella spp. METHODS: It was performed analytical isoelectric focusing of beta-lactamases produced by a group of 33 strains selected in basis of their resistance phenotype. Plasmid profile analysis and assays of transferable drug resistance were developed. The study was completed by hybridization experiments with an intragenic TEM probe which allowed the location of the bla-TEM gene. RESULTS: By analytical isoelectrofocusing we found that 26 out of the 27 ampicillin-resistant strains produced beta-lactamases with pl 5.4 and/or 5.6 corresponding to TEM-1 and/or TEM-2 type. Analysis of plasmid DNA revealed in almost all strains plasmids ranging in size from 1.1 to 125 Mdal. This plasmids were responsible of the resistance and, moreover, were able to transfer the resistance by conjugation mechanisms. Southern blot analysis detected the gene that code the TEM beta-lactamase at the 125, 8 and 5.8 Mdal plasmids. CONCLUSIONS: Resistance to ampicillin in the strains of Salmonella studied was due to the presence of TEM type beta-lactamases coded by conjugative plasmids. These plasmids coded also resistance to other antimicrobial agents. Our results showed that the use of a DNA probe to the detect TEM-type beta-lactamases using a non radioactive probe, could be a suitable alternative to isoelectric focusing.


Assuntos
Resistência a Ampicilina/genética , Proteínas de Bactérias/genética , Salmonella/genética , beta-Lactamases/genética , Proteínas de Bactérias/isolamento & purificação , Southern Blotting , Conjugação Genética , Sondas de DNA , Genes Bacterianos , Humanos , Focalização Isoelétrica , Fatores R , Salmonella/efeitos dos fármacos , Salmonella/isolamento & purificação , Infecções por Salmonella/microbiologia , beta-Lactamases/isolamento & purificação
20.
Eur J Clin Microbiol Infect Dis ; 11(2): 157-61, 1992 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-1327786

RESUMO

To investigate the association between human immunodeficiency virus (HIV) and Epstein-Barr virus (EBV), simultaneous determinations of HIV antigen (HIV Ag) p24 and EBV DNA were performed in lymphocyte culture supernatants from 63 individuals at risk of HIV infection. In vitro data, together with HIV immune status results, were subjected to a statistical analysis. HIV infection was identified in 49 patients (78%); of these, in vitro EBV DNA was found in 44 individuals (90%), while in only 3 of the 14 non-infected ones (21%). Statistical analysis demonstrated a close relationship between evidence of HIV infection and in vitro detection of EBV DNA (87.3% concordant with 95% confidence interval: 76.5%-94.5%). Furthermore, a strong dependence was revealed between the presence of EBV DNA and HIV Ag in culture (p less than 0.00001). These results indicate the existence of in vitro viral interactions, with likely in vivo implications in the pathogenesis and evolution of HIV infection.


Assuntos
DNA Viral/sangue , Proteína do Núcleo p24 do HIV/sangue , Infecções por HIV/microbiologia , HIV-1/imunologia , Herpesvirus Humano 4/isolamento & purificação , Complexo Relacionado com a AIDS/microbiologia , Síndrome da Imunodeficiência Adquirida/microbiologia , Adolescente , Adulto , Western Blotting , Criança , Pré-Escolar , Feminino , Anticorpos Anti-HIV/sangue , Antígenos HIV/sangue , Herpesvirus Humano 4/genética , Humanos , Técnicas Imunoenzimáticas , Lactente , Masculino , Pessoa de Meia-Idade , Hibridização de Ácido Nucleico
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