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1.
Xenotransplantation ; 27(5): e12617, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32557876

RESUMO

The use of decellularized xenogeneic heart valves might offer a solution to overcome the issue of human valve shortage. The aim of this study was to revise decellularization protocols in combination with enzymatic deglycosylation, in order to reduce the immunogenicity of porcine pulmonary heart valves, in means of cells, carbohydrates, and, primarily, Galα1-3Gal (α-Gal) epitope removal. In particular, the valves were decellularized with sodium dodecylsulfate/sodium deoxycholate (SDS/SD), Triton X-100 + SDS (Tx + SDS), or Trypsin + Triton X-100 (Tryp + Tx) followed by enzymatic digestion with PNGaseF, Endoglycosidase H, or O-glycosidase combined with Neuraminidase. Results showed that decellularization alone reduced carbohydrate structures only to a limited extent, and it did not result in an α-Gal free scaffold. Nevertheless, decellularization with Tryp + Tx represented the most effective decellularization protocol in means of carbohydrates reduction. Overall, carbohydrates and α-Gal removal could strongly be improved by applying PNGaseF, in particular in combination with Tryp + Tx treatment, contrary to Endoglycosidase H and O-glycosidase treatments. Furthermore, decellularization with PNGaseF did not affect biomechanical stability, in comparison with decellularization alone, as shown by burst pressure and uniaxial tensile tests. In conclusion, valves decellularized with Tryp + Tx and PNGaseF resulted in prostheses with potentially reduced immunogenicity and maintained mechanical stability.


Assuntos
Bioprótese , Próteses Valvulares Cardíacas , Transplante Heterólogo , Animais , Carboidratos , Glicosilação , Valvas Cardíacas , Humanos , Suínos , Engenharia Tecidual
2.
J Clin Med ; 11(11)2022 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-35683369

RESUMO

The diagnosis of periprosthetic infections (PJI) can be challenging in clinical practice because the clinical presentations of aseptic loosening and low-grade infections are similar. Semiquantitative evaluation of leukocyte esterase (LE) in synovial fluid using a urine strip test has already established itself as a diagnostic method over the past decade. The analysis of LE in synovial fluid leads to a high number of false-positive test results. In the present study, the value of a combined semiquantitative determination of glucose and LE in synovial fluid to improve the diagnosis of PJI was investigated. Over a 4-year period, 145 synovial samples were collected from patients who developed joint effusion after arthroplasty. LE and glucose test strips were considered as an index test for the diagnosis of PJI. A ++ or +++ LE and a negative glucose test strip reading were considered as positive test results. Modified diagnostic criteria for PJI as recommended by the Musculoskeletal Infection Society (MSIS) served as the reference test, except that intraoperative findings were excluded. Forty-six out of 145 samples were classified as septic complication according to the reference test. In regard to PJI, our data showed that combined use of LE and glucose strip test reading displayed a 98.0% specificity (95% confidence interval (CI): 95.2% to 100%), a 50% sensitivity (95% CI: 35.6% to 64.4%), a 92% positive predictive value (95% CI: 81.4% to 100.0%), and an 80.3% negative predictive value (95% CI: 73.2% to 87.4%). In contrast, the exclusive analysis of LE on the urine strip to diagnose PJI demonstrated a 90.9% specificity (95% CI: 85.2% to 96.6%), a 67.4% sensitivity (95% CI: 53.8% to 80.9%), a 77.5% positive predictive value (95% CI: 64.6% to 90.4%), and an 85.7% negative predictive value (5% CI: 79.0% to 92.4%). A combination of LE and glucose test pad reading is considered superior as a potential "rule-in" method for the diagnosis of PJI compared with LE test pad analysis alone. However, combined LE and glucose synovial fluid testing also demonstrated lower test sensitivity and thus diagnostic accuracy compared with LE analysis alone. Therefore, combined glucose and LE test pad analysis does not represent a sufficient diagnostic standard to exclude PJI with certainty.

3.
In Vivo ; 35(3): 1625-1632, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33910845

RESUMO

BACKGROUND/AIM: Septic arthritis (SA) requires rapid diagnosis and therapy to avoid joint damage. This study evaluated the diagnostic accuracy of leukocyte esterase (LE) and glucose (GLC) strip tests for diagnosing SA. PATIENTS AND METHODS: Synovial fluids from 455 patients with atraumatic joint effusions were assessed prospectively over a 5-year period with LE and glucose strip tests. Results were compared to modified Newman criteria for diagnosing joint infections. Synovial fluid cultures, crystal, blood and synovial cell analyses were also performed. RESULTS: Forty-one patients had SA and 252 non-SA. A positive LE reading combined with negative glucose reading could detect SA with 100% specificity, 85% sensitivity, 100% positive predictive value (PPV) and 98% negative predictive value (NPV). Positive synovial LE reading alone detected SA with 82% specificity, 95% sensitivity, 47% PPV, and 99% NPV. CONCLUSION: Combined LE and glucose strip tests represent a low-cost tool for rapidly diagnosing or ruling out SA.


Assuntos
Artrite Infecciosa , Artrite Infecciosa/diagnóstico , Biomarcadores , Hidrolases de Éster Carboxílico , Glucose , Humanos , Indicadores e Reagentes , Valor Preditivo dos Testes , Sensibilidade e Especificidade , Líquido Sinovial
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