Increased Protein Stability and Interleukin-2 Production of a LAT(G131D)Variant With Possible Implications for T Cell Anergy

The adaptor LAT plays a crucial role in the transduction of signals coming from the TCR/CD3 complex. Phosphorylation of some of its tyrosines generates recruitment sites for other cytosolic signaling molecules. Tyrosine 132 in human LAT is essential for PLC-gamma activation and calcium influx generation. It has been recently reported that a conserved glycine residue preceding tyrosine 132 decreases its phosphorylation kinetics, which constitutes a mechanism for ligand discrimination. Here we confirm that a LAT mutant in which glycine 131 has been substituted by an aspartate (LAT(G131D)) increases phosphorylation of Tyr132, PLC-gamma activation and calcium influx generation. Interestingly, the LAT(G131D)mutant has a slower protein turnover while being equally sensitive to Fas-mediated protein cleavage by caspases. Moreover, J.CaM2 cells expressing LAT(G131D)secrete greater amounts of interleukin-2 (IL-2) in response to CD3/CD28 engagement. However, despite this increased IL-2 secretion, J.CaM2 cells expressing the LAT(G131D)mutant are more sensitive to inhibition of IL-2 production by pre-treatment with anti-CD3, which points to a possible role of this residue in the generation of anergy. Our results suggest that the increased kinetics of LAT Tyr132 phosphorylation could contribute to the establishment of T cell anergy, and thus constitutes an earliest known intracellular event responsible for the induction of peripheral tolerance

Health‑related quality of life in patients newly diagnosed with prostate cancer: CAPLIFE study

Funding Funding for open access charge: Universidad de Granada / CBUA. This research was funded by Regional Ministry of Health and Families of Andalusia/Consejería de Salud y Familias de la Junta de Andalucía (PI-0514-2016).Purpose To analyse the Health-Related Quality of Life (HRQoL) at diagnosis of patients with prostate cancer (PCa) according to tumour extension and urinary symptomatology and to explore factors associated with HRQoL. Methods 408 Controls and 463 PCa cases were included. Eligibility criteria were a new diagnosis of PCa (cases), 40–80 years of age, and residence in the participating hospitals’ coverage area for ≥ 6 months before recruitment. HRQoL was evaluated using the 12-Item Short-Form Health Survey, Mental (MCS) and Physical Component Summaries (PCS), and urinary symptoms with the International Prostate Symptom Score. HRQoL scores for all PCa cases, according to tumour extension and urinary symptoms, were compared with controls. In addition, information about lifestyles and comorbidities was collected and its association with low HRQoL (lower scores) were explored using logistic regression models. Results Overall cases had similar PCS score, but lower MCS score than controls. The lowest standardised scores for both PCS and MCS were reached by cases with severe urinary symptoms and a metastatic tumour [mean (SD); PCS: 41.9 (11.5), MCS: 42.3 (10.3)]. Having “below” PCS and MCS scores was associated with the presence of three or more comorbidities in the cases [aOR = 2.86 (1.19–6.84) for PCS and aOR = 3.58 (1.37–9.31) for MCS] and with severe urinary symptomatology [aOR = 4.71 (1.84–12.08) for PCS and aOR = 7.63 (2.70–21.58) for MCS]. Conclusion The mental dimension of HRQoL at diagnosis of patients with PCa was lower than in controls, especially for cases with severe urinary symptoms and a metastatic tumour. Comorbidities and urinary symptoms were variables associated with the HRQoL of PCa cases.Funding for open access charge: Universidad de Granada/CBUARegional Ministry of Health and Families of Andalusia/Consejería de Salud y Familias de la Junta de Andalucía (PI-0514-2016

A Stretch of Negatively Charged Amino Acids of Linker for Activation of T-Cell Adaptor Has a Dual Role in T-Cell Antigen Receptor Intracellular Signaling

The adaptor protein linker for activation of T cells (LAT) has an essential role transducing activatory intracellular signals coming from the TCR/CD3 complex. Previous reports have shown that upon T-cell activation, LAT interacts with the tyrosine kinase Lck, leading to the inhibition of its kinase activity. LAT-Lck interaction seemed to depend on a stretch of negatively charged amino acids in LAT. Here, we have substituted this segment of LAT between amino acids 113 and 126 with a non-charged segment and expressed the mutant LAT (LAT-NIL) in J.CaM2 cells in order to analyze TCR signaling. Substitution of this segment in LAT prevented the activation-induced interaction with Lck. Moreover, cells expressing this mutant form of LAT showed a statistically significant increase of proximal intracellular signals such as phosphorylation of LAT in tyrosine residues 171 and 191, and also enhanced ZAP70 phosphorylation approaching borderline statistical significance (p = 0.051). Nevertheless, downstream signals such as Ca2+ influx or MAPK pathways were partially inhibited. Overall, our data reveal that LAT-Lck interaction constitutes a key element regulating proximal intracellular signals coming from the TCR/CD3 complex.Consejería de Salud de Andalucía, Junta de Andalucía (grants PI-0365-2013 and PI-0055-2017); Instituto de Salud Carlos III (grant PI16-00784 from the “Plan Estatal de I+D+I 2013–2016/FEDER”

Type 2 Diabetes-Related Variants Influence the Risk of Developing Prostate Cancer:A Population-Based Case-Control Study and Meta-Analysis

In this study, we have evaluated whether 57 genome-wide association studies (GWAS)-identified common variants for type 2 diabetes (T2D) influence the risk of developing prostate cancer (PCa) in a population of 304 Caucasian PCa patients and 686 controls. The association of selected single nucleotide polymorphisms (SNPs) with the risk of PCa was validated through meta-analysis of our data with those from the UKBiobank and FinnGen cohorts, but also previously published genetic studies. We also evaluated whether T2D SNPs associated with PCa risk could influence host immune responses by analysing their correlation with absolute numbers of 91 blood-derived cell populations and circulating levels of 103 immunological proteins and 7 steroid hormones. We also investigated the correlation of the most interesting SNPs with cytokine levels after in vitro stimulation of whole blood, peripheral mononuclear cells (PBMCs), and monocyte-derived macrophages with LPS, PHA, Pam3Cys, and Staphylococcus Aureus. The meta-analysis of our data with those from six large cohorts confirmed that each copy of the FTOrs9939609A, HNF1Brs7501939T, HNF1Brs757210T, HNF1Brs4430796G, and JAZF1rs10486567A alleles significantly decreased risk of developing PCa (p = 3.70 × 10−5, p = 9.39 × 10−54, p = 5.04 × 10−54, p = 1.19 × 10−71, and p = 1.66 × 10−18, respectively). Although it was not statistically significant after correction for multiple testing, we also found that the NOTCH2rs10923931T and RBMS1rs7593730 SNPs associated with the risk of developing PCa (p = 8.49 × 10−4 and 0.004). Interestingly, we found that the protective effect attributed to the HFN1B locus could be mediated by the SULT1A1 protein (p = 0.00030), an arylsulfotransferase that catalyzes the sulfate conjugation of many hormones, neurotransmitters, drugs, and xenobiotic com-pounds. In addition to these results, eQTL analysis revealed that the HNF1Brs7501939, HNF1Brs757210, HNF1Brs4430796, NOTCH2rs10923931, and RBMS1rs7593730 SNPs influence the risk of PCa through the modulation of mRNA levels of their respective genes in whole blood and/or liver. These results confirm that functional TD2-related variants influence the risk of developing PCa, but also highlight the need of additional experiments to validate our functional results in a tumoral tissue context

Communication with patients and the duration of family medicine consultations

Objective: To determine the distribution of consultation times, the factors that determine their length, and their relationship with a more participative, patient-centred consulting style. Design: Cross-sectional multicentre study. Location: Primary Healthcare Centres in Andalusia, Spain. Participants: A total of 119 tutors and family medicine physician residents. Principal measurements: Consultation length and communication with the patient were analysed using the CICCAA scale (Connect, Identify, Understand, Consent, Help) during 436 interviews in Primary Care. Results: The mean duration of consultations was 8.8 min (sd: 3.6). The consultation tended to be longer when the physician had a patient-centred approach (10.37 ± 4.19 min vs 7.54 ± 2.98 min; p = 0.001), and when there was joint decision-making (9.79 ± 3.96 min vs 7.73 ± 3.42 min: p = 0.001). In the multivariable model, longer consultations were associated with obtaining higher scores on the CICAA scale, a wider range of reasons for consultation, whether they came accompanied, in urban centres, and a smaller number of daily visits (r2 = 0.32). There was no correlation between physician or patient gender, or problem type. Conclusion: A more patient centred medical profile, increased shared decision-making, a wider range of reasons for consultation, whether they came accompanied, in urban centres, and less professional pressure all seem to be associated with a longer consultation. Resumen: Objetivo: Conocer la distribución del tiempo de consulta, los factores que determinan su duración y su relación con un estilo de consulta participativo y más centrado en el paciente. Diseño: Estudio descriptivo multicéntrico. Emplazamiento: Centros de salud de Atención Primaria en Andalucía (España). Participantes: En total, 119 tutores y residentes de medicina de familia. Mediciones principales: Se analizó el tiempo de consulta y la comunicación con el paciente mediante la escala Conectar, Identificar y Comprender, Acordar y Ayudar (CICAA) en 436 entrevistas en Atención Primaria. Resultados: La duración media de las consultas fue de 8,8 min (DE: 3,86). La consulta fue más larga cuando el profesional tenía un perfil centrado en el paciente (10,37 ± 4,19 vs. 7,54 ± 2,98 min; p = 0,001) y existía toma de decisiones compartida (9,79 ± 3,96 vs. 7,73 ± 3,42 min; p = 0,001). En el modelo multivariable, una mayor duración de la consulta se relacionó con obtener mejores puntuaciones en la escala CICAA, un mayor número de motivos de consulta, presencia de acompañante, centros urbanos y un menor número de visitas diarias (r2 = 0,32). No hubo relación con el sexo del profesional, del paciente ni con el tipo de problema consultado. Conclusiones: Un perfil médico más centrado en el paciente, mayor toma de decisiones compartida, un mayor número de motivos de consulta, la presencia de acompañante, el ser centros de salud urbanos y una menor presión asistencial se muestran asociados a un mayor tiempo de consulta. Keywords: Time, Communication, Physician–patient relations, Primary health care, Patient-centred care, Palabras clave: Tiempo, Comunicación, Relaciones médico-paciente, Atención primaria de salud, Atención dirigida al pacient

A Stretch of Negatively Charged Amino Acids of Linker for Activation of T-Cell Adaptor Has a Dual Role in T-Cell Antigen Receptor Intracellular Signaling

The adaptor protein linker for activation of T cells (LAT) has an essential role transducing activatory intracellular signals coming from the TCR/CD3 complex. Previous reports have shown that upon T-cell activation, LAT interacts with the tyrosine kinase Lck, leading to the inhibition of its kinase activity. LAT–Lck interaction seemed to depend on a stretch of negatively charged amino acids in LAT. Here, we have substituted this segment of LAT between amino acids 113 and 126 with a non-charged segment and expressed the mutant LAT (LAT-NIL) in J.CaM2 cells in order to analyze TCR signaling. Substitution of this segment in LAT prevented the activation-induced interaction with Lck. Moreover, cells expressing this mutant form of LAT showed a statistically significant increase of proximal intracellular signals such as phosphorylation of LAT in tyrosine residues 171 and 191, and also enhanced ZAP70 phosphorylation approaching borderline statistical significance (p = 0.051). Nevertheless, downstream signals such as Ca2+ influx or MAPK pathways were partially inhibited. Overall, our data reveal that LAT–Lck interaction constitutes a key element regulating proximal intracellular signals coming from the TCR/CD3 complex