The effect of a membrane-mimicking environment on the interactions of Cu2+ with an amyloidogenic fragment of chicken prion protein

Prion proteins (PrP) from different species have the ability to tightly bind Cu2+ ions

Association of kidney disease measures with risk of renal function worsening in patients with type 1 diabetes

Background: Albuminuria has been classically considered a marker of kidney damage progression in diabetic patients and it is routinely assessed to monitor kidney function. However, the role of a mild GFR reduction on the development of stage 653 CKD has been less explored in type 1 diabetes mellitus (T1DM) patients. Aim of the present study was to evaluate the prognostic role of kidney disease measures, namely albuminuria and reduced GFR, on the development of stage 653 CKD in a large cohort of patients affected by T1DM. Methods: A total of 4284 patients affected by T1DM followed-up at 76 diabetes centers participating to the Italian Association of Clinical Diabetologists (Associazione Medici Diabetologi, AMD) initiative constitutes the study population. Urinary albumin excretion (ACR) and estimated GFR (eGFR) were retrieved and analyzed. The incidence of stage 653 CKD (eGFR < 60 mL/min/1.73 m2) or eGFR reduction > 30% from baseline was evaluated. Results: The mean estimated GFR was 98 \ub1 17 mL/min/1.73m2 and the proportion of patients with albuminuria was 15.3% (n = 654) at baseline. About 8% (n = 337) of patients developed one of the two renal endpoints during the 4-year follow-up period. Age, albuminuria (micro or macro) and baseline eGFR < 90 ml/min/m2 were independent risk factors for stage 653 CKD and renal function worsening. When compared to patients with eGFR > 90 ml/min/1.73m2 and normoalbuminuria, those with albuminuria at baseline had a 1.69 greater risk of reaching stage 3 CKD, while patients with mild eGFR reduction (i.e. eGFR between 90 and 60 mL/min/1.73 m2) show a 3.81 greater risk that rose to 8.24 for those patients with albuminuria and mild eGFR reduction at baseline. Conclusions: Albuminuria and eGFR reduction represent independent risk factors for incident stage 653 CKD in T1DM patients. The simultaneous occurrence of reduced eGFR and albuminuria have a synergistic effect on renal function worsening

Structure, function, involvement in diseases and targeting of 14-3-3 proteins: An Update

14-3-3 is a class of proteins able to interact with a multitude of targets by establishing protein-protein interactions (PPIs). They are usually found in all eukaryotes with a conserved secondary structure and high sequence homology among species. 14-3-3 proteins are involved in many physiological and pathological cellular processes either by triggering or interfering with the activity of specific protein partners. In the last years, the scientific community has collected many evidences on the role played by seven human 14-3-3 isoforms in cancer or neurodegenerative diseases. Indeed, these proteins regulate the molecular mechanisms associated to these diseases by interacting with (i) oncogenic and (ii) pro-apoptotic proteins and (iii) with proteins involved in Parkinson and Alzheimer diseases. The discovery of small molecule modulators of 14-3-3 PPIs could facilitate complete understanding of the physiological role of these proteins, and might offer valuable therapeutic approaches for these critical pathological states

A Joint DFT-kMC Study to Model Ethylene Carbonate Decomposition Reactions : SEI Formation, Growth, and Capacity Loss During Calendar aging of Li-Metal Batteries

International audienceThe solid electrolyte interphase (SEI) is a multistructured thin layer that forms at the anode (e.g., lithium-metal)/electrolyte (e.g., ethylene carbonate EC) interface due to electrolyte reduction. At the initial battery cycles, the SEI protects the electrolyte from further reduction. However, the SEI continues to grow with time, leading to capacity loss and eventually the death of the battery. In this work, we modeled the battery-aging process at storage conditions (calendar aging). We studied EC decomposition reactions using density functional theory (DFT) simulations in the gas-phase in isolation and over the inorganic layer found inside the SEI composed of Li2CO3. We used the values obtained from DFT alongside diffusion coefficients from the literature to explore the temporal evolution of the concentration of the species by kinetic Monte Carlo (kMC) simulations. We found that reactions occurring over Li2CO3 (001) led to a relatively slow SEI growth which is compatible with the general use of carbonate-based solvents in LIBs for protection/passivation purposes. Our simulations over Li2CO3 (001) predict the formation of a multilayered structured SEI. Moreover, our kMC simulations predict the shift from a nonlinear initial behavior to a linear behavior for the capacity loss induced by the formation and growth of the SEI over time which was reported in previous experimental and theoretical studies for lithiated graphite-based batteries. We extended our analysis to the decomposition reactions over the Li2O (111) surface, which could form from the decomposition of Li2CO3. We found that the selectivity of the decomposition reactions strongly depends on the inorganic surface. The main conclusion of this study is to highlight the crucial role played by surface reactions inside the SEI on the nature and selectivity of the decomposition kinetics of EC for the SEI growth

Immunologic Characterization and T cell Receptor Repertoires of Expanded Tumor-infiltrating Lymphocytes in Patients with Renal Cell Carcinoma

The successful use of expanded tumor-infiltrating lymphocytes (TIL) in adoptive TIL therapies has been reported, but the effects of the TIL expansion, immunophenotype, function, and T cell receptor (TCR) repertoire of the infused products relative to the tumor microenvironment (TME) are not well understood. In this study, we analyzed the tumor samples (n = 58) from treatment-naive patients with renal cell carcinoma (RCC), "pre-rapidly expanded" TILs (pre-REP TIL, n= 15) and "rapidly expanded" TILs (REP TIL, n = 25) according to a clinical-grade TIL production protocol, with single-cell RNA (scRNA)+TCR alpha beta-seq (TCR alpha beta sequencing), TCR beta-sequencing (TCR beta-seq), and flow cytometry. REP TILs encompassed a greater abundance of CD4(+) than CD8(+) T cells, with increased LAG-3 and low PD-1 expressions in both CD4(+) and CD8(+) T cell compartments compared with the pre-REP TIL and tumor T cells. The REP protocol preferentially expanded small clones of the CD4(+) phenotype (CD4, IL7R, KLRB1) in the TME, indicating that the largest exhausted T cell clones in the tumor do not expand during the expansion protocol. In addition, by generating a catalog of RCC-associated TCR motifs from >1,000 scRNA+TCR alpha beta-seq and TCR beta-seq RCC, healthy and other cancer sample cohorts, we quantified the RCC-associated TCRs from the expansion protocol. Unlike the low-remaining amount of anti-viral TCRs throughout the expansion, the quantity of the RCC-associated TCRs was high in the tumors and pre-REP TILs but decreased in the REP TILs. Our results provide an in-depth understanding of the origin, phenotype, and TCR specificity of RCC TIL products, paving the way for a more rationalized production of TILs.Significance: TILs are a heterogenous group of immune cells that recognize and attack the tumor, thus are utilized in various clinical trials. In our study, we explored the TILs in patients with kidney cancer by expanding the TILs using a clinical-grade protocol, as well as observed their characteristics and ability to recognize the tumor using in-depth experimental and computational tools.Peer reviewe

FIGURE 2 from Immunologic Characterization and T cell Receptor Repertoires of Expanded Tumor-infiltrating Lymphocytes in Patients with Renal Cell Carcinoma

REP TIL T cells co-cultured with tumor cells show differences in IFNγ and TNFα expressions. A, REP TILs were cocultured with the corresponding tumor cells (n = 10) either for 6 or 48 hours. The cytokine secretion of CD4+ and CD8+ T cells was analyzed with intracellular flow cytometry staining. The expression of IFNγ and TNFα moderately increased in REP TILs after the co-culture with tumor cells without additional T cell stimulation. After 48 hours co-culture, IFNγ and TNFα levels were increased in both CD4+ (0.74% vs. 0.19%, P = 0.044) and CD8+ T cells (0.78% vs. 0.13%, P = 0.025) compared with baseline REP TILs without tumor cell co-culture. No differences were observed at 6 hours co-cultures (Supplementary Fig. S3A). BASELINE_48h = only REP TILs at 48h, CO-CULTURE_48h = cocultured REP TILs without any stimulation at 48 hours. B, The immunophenotype of T cells after co-culture with tumor cells was analyzed with flow cytometry. The expression of LAG-3 moderately decreased in the CD4+ (median 13.55% vs. 1.84%, P = 0.048) and CD8+ T cells (45.55% vs. 22.94%, P = 0.029) compared with REP TIL baseline cells at 6 hours. The same trends were not observed when REP TILs were co-cultured for 48 hours (Supplementary Fig. S3D). Although not significant, the expression of PD-1 expression increased in half of the patients. BASELINE_6h = only REP TILs at 6h, CO-CULTURE_6h co-cultured REP TILs without any stimulation at 6 hours. C, T cell activation potential was assessed by stimulating the cells with anti-CD3 (OKT3), anti-CD28, anti-CD49d antibodies, and IFNγ and TNFα cytokine secretion was measured as described above. When REP TILs were stimulated and co-cultured for 6 hours, an increase in the CD3+ T cell IFNγ and TNFα expressions was observed between unstimulated and stimulated co-cultures (median 0.72% vs. 1.33%, P = 0.098). A moderate increase in CD4+ T cell IFNγ and TNFα expressions was also observed (0.46% 1.089%, P = 0.012), but not in the CD8+ T cells. CO-CULTURE_6h = cocultured REP TILs without any stimulation at 6h, TSTIM_6h = cocultured REP TILs that were T cell stimulated at 6 hours. D, Prolonged co-culture conditions (48 hours) did not lead to increased IFNγ and TNFα production in the CD3+, CD4+, and CD8+ T cell compartments. CO-CULTURE_48h = cocultured REP TILs without any stimulation at 48h, TSTIM_48h = cocultured REP TILs that were T cell stimulated at 48 hours. Non-parametric Wilcoxon matched-pairs signed rank test was used for all co-culture analyses. ns, not significant; *, P P P P < 0.0001.</p

Figure S12 from Immunologic Characterization and T cell Receptor Repertoires of Expanded Tumor-infiltrating Lymphocytes in Patients with Renal Cell Carcinoma

Figure S12 shows the validation of RCC-associated TCR motifs in various cohorts.</p