Perviedade primária assistida através da angioplastia transluminal na cirurgia de revascularização de membros inferiores criticamente isquemiados / Assisted primary patency through transluminal angioplasty in revascularization surgery of critically ischemic lower limbs

Objetivo: Embora a revisão cirúrgica seja o tratamento tradicionalmente utilizado em lesões estenóticas, a angioplastia transluminal tem-se mostrado uma alternativa menos invasiva para o tratamento dessas lesões. O objetivo deste estudo é analisar os resultados de perviedade obtidos após a angioplastia transluminal de lesões estenóticas pós-operatórias.Método: Foram estudadas prospectivamente 19 angioplastias transluminais realizadas em 16 pacientes com lesões estenóticas diagnosticadas no trajeto do enxerto e consideradas adequadas para a realização do procedimento. As variáveis analisadas foram: tempo de pós-operatório; método diagnóstico; características da estenose; método para realização da angioplastia; sucesso imediato do procedimento; complicações; perviedade em médio prazo.Resultados: A análise das 19 lesões estenóticas favoráveis à angioplastia evidenciou que o tempo médio de pós-operatório foi de 10,26 meses; o eco-Doppler colorido foi o responsável pelo diagnóstico em 68,4 por cento dos casos, todos assintomáticos. Os sítio de angioplastia transluminal foram: corpo do enxerto(terço proximal e terço distal); anastomose distal; anastomose proximal;artéria ilíaca(leito proximal); e leito distal. Após 15 meses, 15 dos 16 pacientes (93,75 por cento) evoluíram sem sintomas isquêmicos. Foram alcançados índices de perviedade primária de 78,9 por cento, de perviedade assistida de 94 por cento, e de salvamento de membro de 100 por cento.Conclusão: A angioplastia transluminal representa um método alternativo e menos invasivo para a manutenção da perviedade e salvamento de membro nos pacientes submetidos à revascularização de membros inferiores...

Anomalies in species identification of enterococci from veterinary sources using a commercial biochemical identification system.

AIMS: A commercial biochemical panel ID kit was used to identify presumptive enterococci isolates of veterinary or agricultural origin obtained during different steps of culture. METHODS AND RESULTS: Fifty isolates identified as enterococci using a genus PCR assay were tested for genus and species identification using the BBL Crystal Identification Gram-Positive ID kit (Becton Dickinson, Sparks, MD, USA). Following sub-culture of the isolates three times, 59% agreement with the original panel ID was obtained. After four and six sub-cultures, percentage agreement increased to 61 and 64%, respectively. Nineteen of the 50 cultures were identified as both Enterococcus faecalis and E. faecium. CONCLUSIONS: As a result of the variability between speciation of isolates following re-culture, additional methods for speciation are warranted. SIGNIFICANCE AND IMPACT OF THE STUDY: This study suggests that the identification of the genus and species of non-human enterococcal isolates can vary greatly during successive passages when using this kit.

Fluoroscopia perioperatória em revascularização de membros inferiores / Perioperative fluoroscopy in lowter lim revascularization

No período de feveretro/1997 a março/2001 a fluoroscopia perioperatoria foi utiliZada em 88 cirurgías de revascularização de membros inferiores. O método foi aplicado no pré-operatório em 14,7 por cento , em 27,2 por cento no intra-operatório e em 97,7 por cento no pós-operatório. Ela determinou alteração de conduta em 25 por cento destas cirurgias.Auxiliou na escolha do leito vascular revascularizado em 6,8 por cento dos casos. No intra-operatório, fatores de risco para a oclusão do enxerto foram identificados e corrigidos em 5,7 por cento. Ao término da círurgia a fluoroscopia determínou reoperação em 12,5 por cento dos casos.A perviedade do enxerto foi de 91,4 por cento e de 89,7 por cento no primeiro e terceiro mês de acompanhamento.A fluoroscopia mostra-se importante método auxiliar à cirurgia de revascularização de membros inferiores. Neste trabalho, sugere sensibilidade superior à arteriografiaconvencional na escolha do leito arterial e na identificação e correção de falhas técnicas que possam comprometer a perviedade do enxerto...

Increased expression of Borrelia burgdorferi vlsE in response to human endothelial cell membranes.

RNA isolated from virulent Borrelia burgdorferi cells incubated with human endothelial or neurological tissue cells was subjected to subtractive hybridization using RNA from the same strain incubated in tissue culture medium alone. This RNA subtractive technique generated specific probes that hybridized to two restriction fragments (8.2 kb and 10 kb respectively) generated by EcoRI digestion of total plasmid DNA. The 10 kb EcoRI fragment localized to lp28-1 and was subsequently identified as the variable membrane protein-like sequence (vls) region, which includes an expression locus (vlsE) and 15 silent cassettes. vlsE encodes a 36 kDa outer surface protein that undergoes antigenic variation during animal infections. Primer extension analysis identified the 5' end of a transcript and a putative promoter for vlsE. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) suggested that the expression of vlsE increased when virulent B. burgdorferi cells were incubated with human tissue cells or purified cell membranes isolated from those same cell lines. A 138 bp region upstream of the vlsE region that was not reported in the genome sequence was sequenced using specific 32P end-labelled primers in a DNA cycle sequencing system at high annealing temperatures. Analysis revealed that it contained a 51 bp inverted repeat, which could form an extremely stable cruciform structure. Southern blots probed with the vlsE promoter/operator region indicated that part or all of this sequence could be found on other B. burgdorferi plasmids.

Dynamics of Salmonella contamination in a commercial quail operation.

Control of carcass contamination requires knowledge of the source and dynamics of spread of Salmonella in commercial poultry production. We examined Salmonella contamination at a U.S. commercial quail operation. Pulsed-field gel electrophoresis (PFGE) was used to type isolates in order to trace Salmonella throughout this production environment. During a 6-mo survey, Salmonella serotypes hadar, typhimurium, typhimurium variant Copenhagen, and paratyphi were encountered within this poultry operation. Ninety-four percent of the Salmonella isolated from breeder and production houses and from carcass rinses belonged to Salmonella serotypes typhimurium variant Copenhagen and hadar. There were six distinct S. typhimurium variant Copenhagen genetic types, as identified by PFGE, present within this particular poultry operation. Seventy-nine percent of S. typhimurium variant Copenhagen identified from the environment of the breeder and production houses produced the same PFGE pattern. Thirty-eight percent of S. typhimurium Copenhagen isolated from carcass rinses and the breeder house shared the same PFGE DNA pattern. This study demonstrates the transmission of salmonellae throughout this production environment, from the breeders to their progeny and to the birds ultimately processed for human consumption.

Determination of close genetic relatedness of the major Salmonella enteritidis phage types by pulsed-field gel electrophoresis and DNA sequence analysis of several Salmonella virulence genes.

Salmonella enteritidis (SE) is an important cause of egg-associated outbreaks in both Europe and the United States. Phage typing has become an important epidemiologic tool in identifying the source of outbreaks. Limitations of phage typing have become apparent with wholesale egg distributors that have multiple suppliers in an area where a particular phage type is endemic. Several different molecular typing methods were evaluated for their discriminatory power to identify genetic differences among different SE phage types isolated in Europe and the United States. Pulsed-field gel electrophoresis (PFGE) identified a single DNA pattern among the different SE phage types. Comparison of the nucleotide sequence for several Salmonella virulence genes failed to identify a single nucleotide change in the gene sequences from most SE isolates, regardless of phage type. On the basis of these results, the different SE phage types appear to be genetically related or clonal. However, with primers 1283 and Opa4, it was possible to differentiate not only SE isolates from different geographic locations but those within a specific geographic locale as well by random amplified polymorphic DNA polymerase chain reaction. Any chance for discerning genetic differences among isolates will need to rely on molecular techniques other than PFGE.

Genetic relatedness of Salmonella isolates from nondomestic birds in Southeastern United States.

Salmonella infections have been implicated in large-scale die-offs of wild birds in the United States. Although we know quite a bit about the epidemiology of Salmonella infection among domestic fowl, we know little about the incidence, epidemiology, and genetic relatedness of salmonellae in nondomestic birds. To gain further insight into salmonellae in these hosts, 22 Salmonella isolates from diseased nondomestic birds were screened for the presence of virulence and antibiotic resistance-associated genes and compared genetically using pulsed-field gel electrophoresis (PFGE) and random amplified polymorphic DNA analysis. Of the 22 Salmonella isolates examined, 15 were positive for the invasion gene invA and the virulence plasmid-associated genes spvC and pef. Most (15 of 22) were generally sensitive to antibiotics. However, two Salmonella isolates from pet birds were identified as Salmonella enterica serovar Typhimurium DT104. Despite the general susceptibility of these Salmonella isolates to most antimicrobial agents, antibiotic resistance-associated genes intI1, merA, and aadA1 were identified in a number of these isolates. Five distinct XbaI and nine distinct BlnI DNA patterns were observed for the 22 Salmonella isolates typed by PFGE. PFGE analysis determined that Salmonella isolates from passerines in Georgia and Wyoming were genetically related.

Radioimmunoscintigraphy with technetium-99m-labelled monoclonal antibody, SM3, in gynaecological cancer.

Radioimmunoscintigraphy (RIS) with technetium-99m labelled SM3, a monoclonal antibody reacting with a polymorphic epithelial mucin glycoprotein core antigen, is evaluated. No adverse effects or thyroid uptake were observed. Studies in 45 patients (one twice) had a sensitivity for gynaecological malignancy of 100% (35/35) and a specificity of 73% (8/11), giving an overall accuracy of 93% (43/46). These results have led to the routine adoption of 99mTc RIS in the management of patients suspected of or having primary or recurrent gynaecological cancer.

Production and use of rat monoclonal antibodies to the human myeloid cell nuclear differentiation antigen.

A dot immunoblot screening assay was used to identify rat monoclonal antibodies to a human myeloid cell differentiation-specific nuclear antigen (MNDA). The selection was based on the positive reaction of hybridoma cell supernatants with a concentrated nuclear protein extract prepared from late stage human myeloid leukemia cells that express MNDA (HL-60) coincident with a negative reaction with the same extract prepared from a non-expressing more immature human myeloid leukemia cell line. The approach provided an efficient method for obtaining monoclonal antibodies to a specific low abundance nuclear antigen that has not been purified. Sixteen wells from three fusions contained antibody displaying a specific reaction with the nuclear protein fraction obtained from the HL-60 cells. Immunoblotting analysis revealed that all of the sixteen specific hybridoma cell lines produced antibody against the same Mr 55,000 nuclear antigen. Selecting hybridoma cells that produce antibody reactive with the native antigen provided antibody suitable for detecting MNDA in immunocytochemical tests. The rat monoclonal antibodies were purified and coupled to CNBr-activated agarose and carbonyldiimidazole-activated agarose. Although both antibody affinity matrices exhibited the same antigen binding capacities, the matrix prepared using carbonyldiimidazole-activated agarose bound the MNDA with a high level of specificity while the matrix prepared from CNBr-activated agarose bound numerous other nuclear proteins.