Association among Terminally Differentiated T Cells, Frailty, and Dependency in a Group of Cuban Centenarians.

INTRODUCTION: Centenarians are considered a model of successful aging. Cuba exhibits one of the oldest populations in Latin America with more than two thousand centenarians. METHODS: This study aimed to evaluate the immune phenotype of forty-three Cuban centenarians, their clinical characteristics such as comorbidities, frailty, body mass index, and some hemochemical parameters. RESULTS: Centenarians had normal body mass indexes, relatively good health status, and 21.95% of them had no comorbidities; 53.6% were classified as frail, and 7% were classified as robust. In addition, 17% of centenarians were independent, and 41.46% were moderately dependent. The seroprevalence against cytomegalovirus was 100%. Concerning pro-inflammatory markers, the majority of them had very low cytokine levels and serum C-reactive protein around the normal limit. We also found the predominance of memory subsets over naive compartments in CD4+ and CD8+ T cells. Terminally differentiated CD8+CD28- T cells were higher in frail centenarians than in pre-frail, while CD8+CD57+ and CD8+EMRA T cells were higher in moderately and severely dependent individuals than in independent individuals. Severely dependent centenarians had a lower CD4+/CD8+ ratio. CONCLUSION: This study describes for the first time the predominance of memory subsets over naive compartments in CD4+ and CD8+ T cells, as well as its relation to frailty and/or dependency in a group of Cuban centenarians. Further studies are needed to continue understanding the natural biological aging mechanism and the relationship between terminally differentiated lymphocytes and inflammaging in the context of extreme longevity.

Combination therapy of Epidermal Growth Factor and Growth Hormone-Releasing Hexapeptide in acute ischemic stroke: a phase I/II non-blinded, randomized clinical trial.

Objective: This study tested the hypothesis that a neuroprotective combined therapy based on epidermal growth factor (EGF) and growth hormone-releasing hexapeptide (GHRP6) could be safe for acute ischemic stroke patients, admitting up to 30% of serious adverse events (SAE) with proven causality. Methods: A multi-centric, randomized, open-label, controlled, phase I-II clinical trial with parallel groups was conducted (July 2017 to January 2018). Patients aged 18-80 years with a computed tomography-confirmed ischemic stroke and less than 12 h from the onset of symptoms were randomly assigned to the study groups I (75 µg rEGF + 3.5 mg GHRP6 i.v., n=10), II (75 µg rEGF + 5 mg GHRP6 i.v., n=10), or III (standard care control, n=16). Combined therapy was given BID for 7 days. The primary endpoint was safety over 6 months. Secondary endpoints included neurological (NIHSS) and functional [Barthel index and modified Rankin scale (mRS)] outcomes. Results: The study population had a mean age of 66 ± 11 years, with 21 men (58.3%), a baseline median NIHSS score of 9 (95% CI: 8-11), and a mean time to treatment of 7.3 ± 2.8 h. Analyses were conducted on an intention-to-treat basis. SAEs were reported in 9 of 16 (56.2%) patients in the control group, 3 of 10 (30%) patients in Group I (odds ratio (OR): 0.33; 95% CI: 0.06-1.78), and 2 of 10 (20%) patients in Group II (OR: 0.19; 95% CI: 0.03-1.22); only two events in one patient in Group I were attributed to the intervention treatment. Compliance with the study hypothesis was greater than 0.90 in each group. Patients treated with EGF + GHRP6 had a favorable neurological and functional evolution at both 90 and 180 days, as evidenced by the inferential analysis of NIHSS, Barthel, and mRS and by their moderate to strong effect size. At 6 months, proportion analysis evidenced a higher survival rate for patients treated with the combined therapy. Ancillary analysis including merged treated groups and utility-weighted mRS also showed a benefit of this combined therapy. Conclusion: EGF + GHRP6 therapy was safe. The functional benefits of treatment in this study supported a Phase III study. Clinical Trial Registration: RPCEC00000214 of the Cuban Public Registry of Clinical Trials, Unique identifier: IG/CIGB-845I/IC/1601.

Glycogen synthase kinase 3 in the world of cell migration.

Glycogen synthase kinase 3 (GSK3) is one of the few master switch kinases that regulate many aspects of cell functions. Recent studies on cell polarization and migration have shown that GSK3 is also essential for proper regulation of these processes. GSK3 influences cell migration as one of the regulators of the spatiotemporally controlled dynamics of the actin cytoskeleton, microtubules, and cell-to-matrix adhesions. In this mini-review, the effects of GSK3 on these three aspects of cell migration will be discussed.

MPL1, a novel phosphatase with leucine-rich repeats, is essential for proper ERK2 phosphorylation and cell motility.

The novel Dictyostelium phosphatase MPL1 contains six leucine-rich repeats at the amino-terminal end and a phosphatase domain at the carboxyl end. Similarly architectured phosphatases exist among other protozoa, such as Entamoeba histolytica, Leishmania major, and Trypanosoma cruzi. MPL1 was strongly induced after 5 h of development; ablation by homologous recombination led to defective streaming and aggregation during development. In addition, cyclic AMP (cAMP)-pulsed mpl1(-) cells showed reduced random and directional motility. At the molecular level, mpl1(-) cells displayed higher prestimulus and persistent poststimulus ERK2 phosphorylation in response to cAMP stimulation. Consistent with their phenotype of persistent ERK2 phosphorylation, mpl1(-) cells also displayed an aberrant pattern of cAMP production, resembling that of the regA(-) cells. Reintroduction of a full-length MPL1 into mpl1(-) cells restored aggregation, ERK2 regulation, random and directional motility, and cAMP production similar to wild-type cells. We propose that MPL1 is a novel phosphatase essential for proper regulation of ERK2 phosphorylation and optimal motility during development.

Comportamiento de variables epidemiológicas y clínicas de pacientes con infarto cerebral en Villa Clara

Fundamento la enfermedad cerebrovascular representa el problema de salud más frecuente relacionado con la atención neurológica, hecho en que estriba la importancia que reviste su estudio en los diferentes contextos y desde diversos enfoques. Objetivo describir el comportamiento de variables epidemiológicas y clínicas en pacientes ingresados por infarto cerebral. Métodos estudio descriptivo y transversal, realizado en el servicio de Neurología del Hospital Arnaldo Milián Castro, de Villa Clara, Cuba, el cual incluyó a todos los pacientes con diagnóstico clínico de infarto cerebral cardioembólico o aterotrombótico, ingresados en sala durante el año 2019. La información se obtuvo de las historias clínicas almacenadas en el Archivo del Hospital; y fue procesada en el paquete estadístico SPSS. v. 21. Se aplicó un análisis estadístico descriptivo, en una distribución de frecuencias. Resultados predominaron los pacientes del sexo femenino (51,6 %). Hubo mayor incidencia en hombres mayores de 79 años (47,7 %), y en mujeres mayores de 70 (86,0 %). En el 67,3 % se demostró la causa cardioembólica. La hipertensión arterial resultó el principal factor de riesgo asociado (83,6 %). El defecto motor se observó como hallazgo clínico más frecuente al ingreso (96,7 %). Se identificaron la transformación hemorrágica del infarto y la bronconeumonía nosocomial como principales complicaciones neurológicas y no neurológicas respectivamente. Prevalecieron los pacientes egresados vivos (68,6 %). Conclusiones los ictus isquémicos son más frecuentes en pacientes de edad avanzada; la identificación temprana y manejo oportuno de la enfermedad instaurada puede prevenir en gran medida la aparición de complicaciones, y consecuentemente la muerte.

Background cerebrovascular disease represents the most common health problem related to neurological care, it is important to study it in different contexts and from different approaches. Objective to describe epidemiological and clinical variables' behavior in patients admitted for stroke. Methods descriptive and cross-sectional study, carried out in Arnaldo Milián Castro Hospital's Neurology service from Villa Clara, Cuba, which included all patients with a clinical diagnosis of cardioembolic or atherothrombotic stroke, admitted to the ward during 2019. The information was obtained from the medical records stored in the Hospital Archive; and it was processed in the statistical package SPSS. v. 21. A descriptive statistical analysis was applied, in a frequency distribution. Results female patients predominated (51.6%). There was a higher incidence in men older than 79 years (47.7%), and in women older than 70 (86.0%). In 67.3% the cardioembolic cause was demonstrated. Arterial hypertension was the main associated risk factor (83.6%). The motor defect was observed as the most frequent clinical finding on admission (96.7%). Hemorrhagic transformation of the infarct and nosocomial bronchopneumonia were identified as the main neurological and non-neurological complications, respectively. Patients discharged alive prevailed (68.6%). Conclusions ischemic strokes are more frequent in elderly patients; early identification and timely management of the established disease can largely prevent the appearance of complications, and consequently death.

Phosphorylation-dependent inhibition of Cdc42 GEF Gef1 by 14-3-3 protein Rad24 spatially regulates Cdc42 GTPase activity and oscillatory dynamics during cell morphogenesis.

Active Cdc42 GTPase, a key regulator of cell polarity, displays oscillatory dynamics that are anticorrelated at the two cell tips in fission yeast. Anticorrelation suggests competition for active Cdc42 or for its effectors. Here we show how 14-3-3 protein Rad24 associates with Cdc42 guanine exchange factor (GEF) Gef1, limiting Gef1 availability to promote Cdc42 activation. Phosphorylation of Gef1 by conserved NDR kinase Orb6 promotes Gef1 binding to Rad24. Loss of Rad24-Gef1 interaction increases Gef1 protein localization and Cdc42 activation at the cell tips and reduces the anticorrelation of active Cdc42 oscillations. Increased Cdc42 activation promotes precocious bipolar growth activation, bypassing the normal requirement for an intact microtubule cytoskeleton and for microtubule-dependent polarity landmark Tea4-PP1. Further, increased Cdc42 activation by Gef1 widens cell diameter and alters tip curvature, countering the effects of Cdc42 GTPase-activating protein Rga4. The respective levels of Gef1 and Rga4 proteins at the membrane define dynamically the growing area at each cell tip. Our findings show how the 14-3-3 protein Rad24 modulates the availability of Cdc42 GEF Gef1, a homologue of mammalian Cdc42 GEF DNMBP/TUBA, to spatially control Cdc42 GTPase activity and promote cell polarization and cell shape emergence.

Principales factores pronósticos, clínicos y epidemiológicos en pacientes con infarto cerebral total de circulación anterior / Main prognostic, clinical and epidemiological factors in patients with total cerebral infarction of previous circulation

RESUMEN Fundamento: El comportamiento epidemiológico del infarto cerebral conlleva a la necesidad de estudios encaminados a caracterizar el pronóstico de los pacientes, y a establecer factores correlacionados con la evolución, resumidos en modelos que permitan dirigir acciones de tratamiento hacia dichos determinantes. Objetivo: describir los principales factores pronósticos, clínicos y epidemiológicos en pacientes con infarto cerebral total de circulación anterior. Métodos: estudio descriptivo, en pacientes adultos (N=35) con infarto cerebral total de circulación anterior, ingresados en la Unidad de Ictus, del Hospital Arnaldo Milián Castro, desde marzo de 2017 hasta marzo de 2018. Se analizaron variables clínico-demográficas y relacionadas con la lesión isquémica, y se asociaron con el pronóstico global de los pacientes. Se aplicaron el Test de independencia basado en la Distribución Chi-cuadrado y la Prueba t de Student. Resultados: el 81,82 % de los pacientes entre 75 y 85 años evolucionó desfavorablemente, y fue mayor la probabilidad de un peor pronóstico en los pacientes más longevos (X2=10,59; p=0,007). En los que presentaron cifras más elevadas de presión arterial media y mayor tiempo con saturación de oxígeno menor que 90 % durante el sueño, la probabilidad de un pronóstico final desfavorable fue mayor (p=0,000). Ninguno de los factores asociados a la lesión isquémica se relacionó significativamente con el pronóstico (p>0,05). Conclusión: la edad, el promedio de presión arterial media, y el tiempo de saturación de oxígeno menor de 90 % durante el sueño en la fase aguda del ictus, se asocian con mayor probabilidad de presentar un pronóstico global desfavorable.

ABSTRACT Foundation: The epidemiological behavior of cerebral infarction leads to the need for studies aimed at characterizing the prognosis of patients, and establishing factors correlated with evolution, summarized in models that allow directing treatment actions towards these determinants. Objective: to describe the main prognostic, clinical and epidemiological factors in patients with total cerebral infarction of anterior circulation. Methods: descriptive study, in adult patients (N=35) with total cerebral infarction of previous circulation, admitted to the Stroke Unit, of the Arnaldo Milián Castro Hospital, from March 2017 to March 2018. Clinical-demographic variables were analyzed and related to ischemic injury, and were associated with the overall prognosis of patients. The Independence Test based on the Chi-square Distribution and the Student t-test were applied. Results: 81.82 % of patients between 75 and 85 years evolved unfavorably, and the probability of a worse prognosis in older patients was higher (X2 = 10.59; p = 0.007). In those who presented higher levels of mean blood pressure and longer time with oxygen saturation less than 90 % during sleep, the probability of an unfavorable final prognosis was higher (p =0.000). None of the factors associated with ischemic injury was significantly related to the prognosis (p>0.05). Conclusion: age, average mean blood pressure, and oxygen saturation time of less than 90 % during sleep in the acute phase of stroke are associated with a higher probability of presenting an unfavorable overall prognosis.

Dictyostelium kinase DPYK3 negatively regulates STATc signaling in cell fate decision.

DPYK3, a member of the Dictyostelium TKL (tyrosine kinase like) kinase family, was ablated by homologous recombination. dpyk3- cells displayed aberrant pattern formation during development. The prestalk O zone was not properly formed and, instead, the prespore zone was expanded in dpyk3- slugs. During development, the transcription factor STATc (signal transducers and activators of transcription c) was persistently phosphorylated and ecmAO expression level was kept low in dpyk3- cells. Furthermore, in response to differentiation inducing factor-1 (DIF-1) in suspension culture, dpyk3- cells displayed persistent STATc phosphorylation and reintroduction of DPYK3 in dpyk3- cells restored transient STATc phosphorylation similarly to wild type cells. In contrast to the positive STAT regulation by Janus Kinase in metazoans, Dictyostelium DPYK3 negatively regulates STATc during development in response to DIF-1 signaling.