Actividad antioxidante de hidrolizados proteicos de caupí (variedad cuarentón) obtenidos por digestión enzimática

El caupí (vigna unguiculata (L.) Walp.) es una leguminosa cultivada en el Nordeste Argentino con un elevado contenido de proteínas (23-26%) de buena calidad nutricional. Constituye una atractiva materia prima de péptidos bioactivos para su incorporación en alimentos. El objetivo fue estudiar la actividad antioxidante de péptidos obtenidos por hidrólisis enzimática a partir de proteínas de caupí. Aislados proteicos se obtuvieron a diferentes pH de extracción pH 8 (A8) y pH 10 (A10). Los hidrolizados fueron generados con alcalasa (≥ 2.4 U/g; 37° C, 4 h) en relación μl enzima/mg muestra: 0,8/100 (bajo grado de hidrólisis: A8LH y A10LH) y 8/100 (alto grado de hidrólisis: A8HH y A10HH). En cuanto al grado de hidrólisis (método TNBS), A8LH presentó un valor significativamente menor respecto a A10LH (2,28 y 4,60%) mientras que A8HH y A10HH alcanzaron un mayor grado de hidrólisis (24 y 26%). Los perfiles electroforéticos (SDS-PAGE-ME 15%) mostraron que en A8LH y A10LH la hidrólisis provocó desaparición de algunos polipéptidos con masas moleculares >20 kDa y un incremento de polipéptidos <20 kDa. En A8HH y A10HH se visualizaron bandas <20 kDa. A8 y A10 presentaron similar actividad antioxidante medida por ABTS·+ (IC50A8=23,54 mg/mL, IC50A10=22,78 mg/mL). La hidrólisis incrementó significativamente la actividad antioxidante con respecto al aislado: los IC50 de A8LH y A8HH fueron un 74% y 83% menor que A8, respectivamente. Un comportamiento similar se observó para A10LH y A10HH respecto a A10. Los resultados obtenidos brindan información relevante, mostrando un aumento de la actividad antioxidante por efecto de la hidrólisis con alcalasa. Posteriores estudios se realizarán por otros mecanismos de neutralización de radicalesFil: Gómez, Andrea Gisella. Universidad Nacional del NordesteFil: Gay, Claudia Carolina. Universidad Nacional del NordesteFil: Avanza, María Victoria. Universidad Nacional del Nordest

Melflufen: A Peptide-Drug Conjugate for the Treatment of Multiple Myeloma

Despite the availability of new therapies that have led to improved outcomes for patients with multiple myeloma, most patients will eventually relapse. With triplet and even quadruplet combination therapies becoming standard in the first and second line, many patients will have few treatment options after second-line treatment. Melflufen (melphalan flufenamide) is a first-in-class peptide-drug conjugate (PDC) that targets aminopeptidases and rapidly releases alkylating agents into tumor cells. Once inside the tumor cells, melflufen is hydrolyzed by peptidases to release alkylator molecules, which become entrapped. Melflufen showed anti-myeloma activity in myeloma cells that were resistant to bortezomib and the alkylator melphalan. In early phase studies (O-12-M1 and HORIZON [OP-106]), melflufen plus dexamethasone has demonstrated encouraging clinical activity and a manageable safety profile in heavily pretreated patients with relapsed/refractory multiple myeloma, including those with triple-class refractory disease and extramedullary disease. The Phase III OCEAN study (OP-104) is further evaluating melflufen plus dexamethasone in patients with relapsed/refractory multiple myeloma. The safety profile of melflufen is characterized primarily by clinically manageable hematologic adverse events. Melflufen, with its novel mechanism of action, has the potential to provide clinically meaningful benefits to patients with relapsed/refractory multiple myeloma, including those with high unmet needs.The authors received medical editorial support for the development of this manuscript, which was funded by Oncopeptides AB

Evaluación in vitro de la actividad proteolítica de células de carcinoma renal de células claras (Caki-1) en condiciones de hipoxia

La proteólisis extracelular es uno de los principales mecanismos por lo que la transformación de la matriz intercelular se desarrolla, y facilita la metástasis tumoral. Las enzimas que la catalizan se denominan metaloproteinasas de la matriz (MMPs); son proteinasas pertenecientes a una amplia familia, que se asocian con tumorigénesis, renovación de la matriz extracelular y la migración de células cancerosas, control del crecimiento, inflamación y angiogénesis. Dados estos roles tan centrales en la biología del cáncer, desde hace tiempo vienen siendo estudiadas, pero son prácticamente inexistentes los trabajos que las relacionan con su expresión en condiciones hipóxicas y más aún en Carcinoma Renal de Células Claras (CRCC). Por tal motivo, en el presente trabajo, se inició la caracterización de la expresión de MMPs secretadas por células Caki-1, modelo celular ampliamente validado para el CRCC. Utilizando un modelo in vitro de cultivo celular se generó hipoxia con el agregado de CoCl2 (100, 200 y 300 μM) y se evaluó si la hipoxia influye en la actividad proteolítica del sobrenadante celular. Se utilizó SDS-PAGE para analizar el perfil proteico y la técnica de zimografía para determinar la actividad gelatinolítica. Asimismo, se estudió la expresión de las MMPs (2, 9 y 13) por Dot-Blott. Los resultados hallados muestran que la hipoxia exacerba la actividad proteolítica de estas células, y que en dicha actividad están implicadas las enzimas MMP-2 y MMP-13, principalmente y en menor grado MMP-9, secretadas al medio extracelular. Actualmente, se está expandiendo la búsqueda de otras isoformas de MMPs (tales como MMP1, MMP8, MMP 21) y evaluando su expresión en el otro modelo validado de CRCC, las células Caki-2Fil: Palomar, Lucas Sebastián. Universidad Nacional del NordesteFil: Melana Colavita, Juan Pablo. Universidad Nacional del NordesteFil: Aguirre, María Victoria. Universidad Nacional del NordesteFil: Gay, Claudia Carolina. Universidad Nacional del NordesteFil: Rodríguez, Juan Pablo. Universidad Nacional del Nordest

2021 European Myeloma Network review and consensus statement on smoldering multiple myeloma: how to distinguish (and manage) Dr. Jekyll and Mr. Hyde

According to the updated International Myeloma Working Group criteria, smoldering multiple myeloma (SMM) is an asymptomatic plasma cell disorder characterized by an M-component >3 g/dL, bone marrow plasma cell infiltration >10% and <60%, and absence of any myeloma-defining event. Active multiple myeloma is preceded by SMM, with a median time to progression of approximately 5 years. Cases of SMM range from the extremes of monoclonal gammopathy of undetermined significance-like, in which patients never progress during their lifetimes, to early multiple myeloma, in which transformation into symptomatic disease, based on genomic evolution, may be rapid and devastating. Such a split personality makes the prognosis and management of individual patients challenging, particularly with regard to the identification and possible early treatment of high-risk SMM. Outside of clinical trials, the conventional approach to SMM generally remains close observation until progression to active multiple myeloma. However, two prospective, randomized trials have recently demonstrated a significant clinical benefit in terms of time to progression, and of overall survival in one of the two studies, for some patients with higher-risk SMM treated with lenalidomide +/- dexamethasone, raising the question of whether such an approach should be considered a new standard of care. In this paper, experts from the European Myeloma Network describe current biological and clinical knowledge on SMM, focusing on novel insights into its molecular pathogenesis, new prognostic scoring systems proposed to identify SMM patients at higher risk of early transformation, and updated results of completed or ongoing clinical trials. Finally, some practical recommendations for the real-life management of these patients, based on Delphi consensus methodology, are provided

Meta-analysis of ciltacabtagene autoleucel versus physician’s choice therapy for the treatment of patients with relapsed or refractory multiple myeloma

[Objective]: In the absence of head-to-head trials, indirect treatment comparisons (ITCs) between ciltacabtagene autoleucel (cilta-cel; in CARTITUDE-1) and treatments used in real-world clinical practice (physician’s choice of treatment [PCT]), were previously conducted. We conducted multiple meta-analyses using available ITC data to consolidate the effectiveness of cilta-cel versus PCT for patients with triple-class exposed relapsed or refractory multiple myeloma (RRMM). [Methods]: Five ITCs were assessed for similarity to ensure robust comparisons using meta-analysis. Effectiveness outcomes were overall survival (OS), progression-free survival (PFS), time to next treatment (TTNT), and overall response rate (ORR). A robust variance estimator was used to account for the use of CARTITUDE-1 in each pairwise ITC. Analyses were conducted in both treated and enrolled populations of CARTITUDE-1. [Results]: Four ITCs were combined for evaluation of OS. Results were statistically significantly in favor of cilta-cel versus PCT in treated patients (hazard ratio [HR]: 0.24, 95% confidence interval [CI]: 0.22–0.26). Three ITCs were combined for evaluation of PFS and TTNT. Cilta-cel reduced the risk of progression and receiving a subsequent treatment by 80% (HR: 0.20 [95% CI: 0.06, 0.70]) and 83% (HR: 0.17 [95% CI: 0.12, 0.26]), respectively. Three ITCs were combined for evaluation of ORR. Cilta-cel increased the odds of achieving an overall response by 86-times versus PCT in treated patients. Findings were consistent in the enrolled populations and across sensitivity analyses. [Conclusions]: Evaluating multiple indirect comparisons, cilta-cel demonstrated a significantly superior advantage over PCT, highlighting its effectiveness as a therapy in patients with triple-class exposed RRMM.The CARTITUDE-1 study and these analyses were funded by Janssen Research & Development, LLC, and Legend Biotech, Inc. Medical writing support was provided by EVERSANA and funded by Janssen Global Services, LLC

Protein flexibility directs DNA recognition by the papillomavirus E2 proteins

Although DNA flexibility is known to play an important role in DNA–protein interactions, the importance of protein flexibility is less well understood. Here, we show that protein dynamics are important in DNA recognition using the well-characterized human papillomavirus (HPV) type 6 E2 protein as a model system. We have compared the DNA binding properties of the HPV 6 E2 DNA binding domain (DBD) and a mutant lacking two C-terminal leucine residues that form part of the hydrophobic core of the protein. Deletion of these residues results in increased specific and non-specific DNA binding and an overall decrease in DNA binding specificity. Using 15N NMR relaxation and hydrogen/deuterium exchange, we demonstrate that the mutation results in increased flexibility within the hydrophobic core and loop regions that orient the DNA binding helices. Stopped-flow kinetic studies indicate that increased flexibility alters DNA binding by increasing initial interactions with DNA but has little or no effect on the structural rearrangements that follow this step. Taken together these data demonstrate that subtle changes in protein dynamics have a major influence on protein–DNA interactions

Comparative Study of Infliximab Versus Adalimumab in Refractory Uveitis Due to Behçet's Disease: National Multicenter Study of 177 Cases

Objective: To compare the efficacy of infliximab (IFX) versus adalimumab (ADA) as a first-line biologic drug over 1 year of treatment in a large series of patients with refractory uveitis due to Behçet's disease (BD). Methods: We conducted an open-label multicenter study of IFX versus ADA for BD-related uveitis refractory to conventional nonbiologic treatment. IFX or ADA was chosen as the first-line biologic agent based on physician and patient agreement. Patients received 3-5 mg/kg intravenous IFX at 0, 2, and 6 weeks and every 4-8 weeks thereafter, or 40 mg subcutaneous ADA every other week without a loading dose. Ocular parameters were compared between the 2 groups. Results: The study included 177 patients (316 affected eyes), of whom 103 received IFX and 74 received ADA. There were no significant baseline differences between treatment groups in main demographic features, previous therapy, or ocular sign severity. After 1 year of therapy, we observed an improvement in all ocular parameters in both groups. However, patients receiving ADA had significantly better outcomes in some parameters, including improvement in anterior chamber inflammation (92.31% versus 78.18% for IFX; P = 0.06), improvement in vitritis (93.33% versus 78.95% for IFX; P = 0.04), and best-corrected visual acuity (mean ± SD 0.81 ± 0.26 versus 0.67 ± 0.34 for IFX; P = 0.001). A nonsignificant difference was seen for macular thickness (mean ± SD 250.62 ± 36.85 for ADA versus 264.89 ± 59.74 for IFX; P = 0.15), and improvement in retinal vasculitis was similar between the 2 groups (95% for ADA versus 97% for IFX; P = 0.28). The drug retention rate was higher in the ADA group (95.24% versus 84.95% for IFX; P = 0.042). Conclusion: Although both IFX and ADA are efficacious in refractory BD-related uveitis, ADA appears to be associated with better outcomes than IFX after 1 year of follow-up