RESUMEN
Protein glycosylation, a critical post-translational modification, influences the stability, efficacy, and immunogenicity of recombinant proteins, including biopharmaceuticals. Glycan structures exhibit significant heterogeneity, varying with production cell types, culture conditions, and purification methods. Consequently, monitoring and evaluating the glycan structures of recombinant proteins is vital, particularly in biopharmaceutical production. The lectin microarray, a technique complementary to mass spectrometry, boasts high sensitivity and ease of use. However, it typically requires more than a day to yield results. To adapt it to non-glycoscience research or drug product process development, an automated, high-throughput alternative is needed. Therefore, the world's first fully automated lectin-based glycan profiling system was developed, utilizing the "bead array in a single tip (BIST)" technology concept. This system allows for the preparation and storage of lectin-immobilized beads in units of 1,000, with customizable parallel insertion orders for various purposes. This article presents a practical protocol for research involving "glyco-qualified" recombinant proteins. After testing their reactivity against 12 polyacrylamide-glycan conjugates, 15 lectins were selected to increase the system's versatility. In addition, the sample labeling process was optimized by switching from Cy3 to biotin, reducing the overall processing time by 30 min. For immediate data qualification, lectin-binding signals are displayed as a dotcode on the top monitor. The system's reliability was confirmed through day-to-day reproducibility tests, repeatability tests, and long-term storage tests, with a coefficient of variation of <10%. This user-friendly and rapid glyco-analyzer has potential applications in the quality monitoring of endogenous glycoproteins for biomarker evaluation and validation. This method facilitates analysis for those new to glycoscience, thereby broadening its practical utility.
Asunto(s)
Lectinas , Polisacáridos , Proteínas Recombinantes , Proteínas Recombinantes/química , Polisacáridos/química , Polisacáridos/análisis , Lectinas/química , Glicosilación , Automatización de Laboratorios/métodosRESUMEN
BACKGROUND: Low muscle mass in patients with acute heart failure (AHF) is associated with poor prognosis; however, this is based on a single baseline measurement, with little information on changes in muscle mass during hospitalization and their clinical implications. This study investigated the relationship between changes in rectus femoris cross-sectional area (RFCSA) on ultrasound and the prognosis of patients with AHF. METHODSâANDâRESULTS: This is a retrospective evaluation of 284 AHF patients (mean [±SD] age 79.1±11.9 years; 116 female). RFCSA assessments at admission (pre-RFCSA), ∆RFCSA (i.e., the percentage change in RFCSA from admission to 2 weeks), and composite prognosis (all-cause death and heart failure-related readmission) within 1 year were determined. Patients were divided into 4 groups according to their median pre-RFCSA and ∆RFCSA after sex stratification: Group A, higher pre-RFCSA/better ∆RFCSA; Group B, higher pre-RFCSA/worse ∆RFCSA; Group C, lower pre-RFCSA/better ∆RFCSA; Group D, lower pre-RFCSA/worse ∆RFCSA. In the Cox regression analysis, with Group A as the reference, the cumulative event rate of Group C (hazard ratio [HR] 3.39; 95% confidence interval [CI] 0.71-16.09; P=0.124) did not differ significantly; however, the cumulative event rates of Group B (HR 7.93; 95% CI 1.99-31.60; P=0.003) and Group D (HR 9.24; 95% CI 2.57-33.26; P<0.001) were significantly higher. CONCLUSIONS: ∆RFCSA during hospitalization is useful for risk assessment of prognosis in patients with AHF.
Asunto(s)
Insuficiencia Cardíaca , Músculo Cuádriceps , Ultrasonografía , Humanos , Insuficiencia Cardíaca/diagnóstico por imagen , Insuficiencia Cardíaca/mortalidad , Femenino , Masculino , Anciano , Estudios Retrospectivos , Anciano de 80 o más Años , Músculo Cuádriceps/diagnóstico por imagen , Pronóstico , Enfermedad Aguda , Readmisión del Paciente/estadística & datos numéricos , Sarcopenia/diagnóstico por imagenRESUMEN
BACKGROUND & AIMS: This study aimed to determine the impact of sarcopenia and nutritional risk on swallowing-related muscles by ultrasonography and dysphagia occurrence in older patients with acute heart failure (AHF) during hospitalization. METHODS: Patients with AHF aged ≥65 years without dysphagia (Food Intake LEVEL Scale [FILS] score ≥9) before admission were classified into four groups at admission: robust group, sarcopenia group (Asian Working Group for Sarcopenia 2019), nutritional risk group (geriatric nutritional risk index <92), and complicated group (with both sarcopenia and nutritional risk). Swallowing function (maximal hyoid displacement, geniohyoid muscle area and brightness, and maximal tongue pressure) and FILS were investigated from the medical records. RESULTS: In total, 131 patients with AHF (mean age 82.8 ± 7.1 years, 71 males) were enrolled during the study period; 33, 58, 5, and 35 were classified into the robust, sarcopenia, nutritional risk, and complicated groups, respectively. In the covariance analysis adjusted for age, sex, comorbidities, and cardiac function, the complicated group had significantly worse swallowing function than the sarcopenia and robust groups (P < 0.05). In the Cox proportional hazards model, in which event occurrence was defined as the first-time FILS score of ≥9 obtained during hospitalization, the sarcopenia group (hazard ratio [HR]: 0.83, 95 % confidence interval [CI]: 0.51-1.34, P = 0.438) and nutritional risk group (HR: 0.77, 95 % CI:0.25-2.32, P = 0.637) were not significantly different, but the complicated group (HR: 0.54, 95 % CI: 0.31-0.95, P = 0.033) had significantly lower cumulative event rates with the robust group as the reference. CONCLUSION: Sarcopenia and nutritional risk in older patients with AHF are risk factors for decreased swallowing function.
Asunto(s)
Trastornos de Deglución , Insuficiencia Cardíaca , Desnutrición , Sarcopenia , Masculino , Humanos , Anciano , Anciano de 80 o más Años , Sarcopenia/complicaciones , Sarcopenia/diagnóstico por imagen , Sarcopenia/epidemiología , Deglución , Trastornos de Deglución/epidemiología , Estudios Retrospectivos , Presión , Lengua , Desnutrición/complicaciones , Insuficiencia Cardíaca/complicaciones , Insuficiencia Cardíaca/diagnóstico por imagen , UltrasonografíaRESUMEN
Electrical muscle stimulation (EMS) is expected to be considered as an add-on therapy for the usual rehabilitation of patients with chronic heart failure (HF). However, it remains unclear whether EMS can reduce muscle volume loss in patients with acute HF (AHF) immediately after hospitalization. Therefore, the aim of this study was to investigate if EMS could reduce the lower-limb muscle volume loss in patients with AHF. In this single-center, retrospective, observational study, lower-limb skeletal muscle volume, quadriceps muscle layer thickness, and clinical events (worsening HF or kidney function) were evaluated in 45 patients with AHF (mean age, 77.4 ± 11.6 years, 31 males). All patients underwent EMS on the right leg, in addition to usual rehabilitation, for 20 minutes per day, 5 days per week, for 2 weeks. A two-factor (time × leg) analysis of variance was performed to compare the difference between the right leg (usual rehabilitation and EMS) and left leg (usual rehabilitation only). The skeletal muscle mass decreased by 11.6% ± 19.7% from baseline in the right leg and by 20.4% ± 16.1% in the left leg (interaction; F = 4.54, P = 0.036). The quadriceps muscle layer thickness decreased by 10.2% ± 7.1% from baseline in the right leg and by 13.5% ± 6.0% in the left leg (interaction; F = 10.50, P = 0.002). No clinical events were related to EMS. Results showed that EMS combined with usual rehabilitation for patients with AHF has the potential to inhibit muscle volume loss.
Asunto(s)
Insuficiencia Cardíaca , Pierna , Masculino , Humanos , Anciano , Anciano de 80 o más Años , Estimulación Eléctrica , Insuficiencia Cardíaca/complicaciones , Insuficiencia Cardíaca/terapia , Extremidad Inferior , Músculo EsqueléticoRESUMEN
Polystyrene-supported TBD (PS-TBD) catalyzes the ring-opening of N-tosylaziridines with silylated nucleophiles to give the corresponding products in high yields. PS-TBD was easily recovered and reused without significant loss of catalytic activity.