SUMOylation regulates LKB1 localization and its oncogenic activity in liver cancer

BACKGROUND: Even though liver kinase B1 (LKB1) is usually described as a tumor suppressor in a wide variety of tissues, it has been shown that LKB1 aberrant expression is associated with bad prognosis in Hepatocellular Carcinoma (HCC). METHODS: Herein we have overexpressed LKB1 in human hepatoma cells and by using histidine pull-down assay we have investigated the role of the hypoxia-related post-translational modification of Small Ubiquitin-related Modifier (SUMO)ylation in the regulation of LKB1 oncogenic role. Molecular modelling between LKB1 and its interactors, involved in regulation of LKB1 nucleocytoplasmic shuttling and LKB1 activity, was performed. Finally, high affinity SUMO binding entities-based technology were used to validate our findings in a pre-clinical mouse model and in clinical HCC. FINDINGS: We found that in human hepatoma cells under hypoxic stress, LKB1 overexpression increases cell viability and aggressiveness in association with changes in LKB1 cellular localization. Moreover, by using site-directed mutagenesis, we have shown that LKB1 is SUMOylated by SUMO-2 at Lys178 hampering LKB1 nucleocytoplasmic shuttling and fueling hepatoma cell growth. Molecular modelling of SUMO modified LKB1 further confirmed steric impedance between SUMOylated LKB1 and the STe20-Related ADaptor cofactor (STRADα), involved in LKB1 export from the nucleus. Finally, we provide evidence that endogenous LKB1 is modified by SUMO in pre-clinical mouse models of HCC and clinical HCC, where LKB1 SUMOylation is higher in fast growing tumors. INTERPRETATION: Overall, SUMO-2 modification of LKB1 at Lys178 mediates LKB1 cellular localization and its oncogenic role in liver cancer. FUND: This work was supported by grants from NIH (US Department of Health and Human services)-R01AR001576-11A1 (J.M.M and M.L.M-C.), Gobierno Vasco-Departamento de Salud 2013111114 (to M.L.M.-C), ELKARTEK 2016, Departamento de Industria del Gobierno Vasco (to M.L.M.-C), MINECO: SAF2017-87301-R and SAF2014-52097-R integrado en el Plan Estatal de Investigación Cientifica y Técnica y Innovación 2013-2016 cofinanciado con Fondos FEDER (to M.L.M.-C and J.M.M., respectively), BFU2015-71017/BMC MINECO/FEDER, EU (to A.D.Q. and I.D.M.), BIOEF (Basque Foundation for Innovation and Health Research): EITB Maratoia BIO15/CA/014; Instituto de Salud Carlos III:PIE14/00031, integrado en el Plan Estatal de Investigación Cientifica y Técnica y Innovacion 2013-2016 cofinanciado con Fondos FEDER (to M.L.M.-C and J.M.M), Asociación Española contra el Cáncer (T.C.D, P·F-T and M.L.M-C), Daniel Alagille award from EASL (to T.C.D), Fundación Científica de la Asociación Española Contra el Cancer (AECC Scientific Foundation) Rare Tumor Calls 2017 (to M.L.M and M.A), La Caixa Foundation Program (to M.L.M), Programma di Ricerca Regione-Università 2007-2009 and 2011-2012, Regione Emilia-Romagna (to E.V.), Ramón Areces Foundation and the Andalusian Government (BIO-198) (A.D.Q. and I.D.M.), ayudas para apoyar grupos de investigación del sistema Universitario Vasco IT971-16 (P.A.), MINECO:SAF2015-64352-R (P.A.), Institut National du Cancer, FRANCE, INCa grant PLBIO16-251 (M.S.R.), MINECO - BFU2016-76872-R to (E.B.). Work produced with the support of a 2017 Leonardo Grant for Researchers and Cultural Creators, BBVA Foundation (M.V-R). Finally, Ciberehd_ISCIII_MINECO is funded by the Instituto de Salud Carlos III. We thank MINECO for the Severo Ochoa Excellence Accreditation to CIC bioGUNE (SEV-2016-0644). Funding sources had no involvement in study design; in the collection, analysis, and interpretation of data; in the writing of the report; and in the decision to submit the paper for publication

Radioactive immunocomplex formation in a double antibody ria

Se ha elaborado un modelo para el estudio de la cinética y equilibrio de las reacciones antígeno-anticuerpo implicadas en el radioinmunoanálisis (RIA) de doble anticuerpo correspondiente al Péptido C. Se pretende caracterizar la influencia de las concentraciones iniciales de antígeno marcado (M) y no marcado (Q) sobre la formación del inmunocomplejo (PM) al segundo anticuerpo (J) inmovilizado sobre una bola. Se realizan 30 experiencias para el estudio del efecto de las variables antes mencionadas. Los resultados obtenidos son concordantes con el modelo propuesto.A model has been produced for the kinetic and equilibrium study of antigen-antibody reactions in the radioimmunoassay (RIA) of C-Peptide double antibody. The aim is (1) to characterise the influence of initial concentrations of labelled (M) and unlabelled (Q) antigen and that of the initial concentration of the antibody in solution (P), and (2) to study the binding of the immunocomplex (PM) to the second antibody (J) immobilised on a bead, and the replacement of M by Q in the PMJ immunocomplex. In order to study the effect of such variables, 44 experiments were conducted. The results are in line with the model proposed

Automated and unobtrusive measurement of physical activity in an interactive playground

© 2019 Promoting physical activity is one of the main goals of interactive playgrounds. To validate whether this goal is met, we need to measure the amount of physical player activity. Traditional methods of measuring activity, such as observations or annotations of game sessions, require time and personnel. Others, such as heart rate monitors and accelerometers, need to be worn by the player. In this paper, we investigate whether physical activity can be measured unobtrusively by tracking players using depth cameras and applying computer vision algorithms. In a user study with 32 players, we measure the players’ speed while playing a game of tag, and demonstrate that our measures correlate well with exertion measured using heart rate sensors. This makes the method an attractive alternative to either manual coding or the use of worn devices. We also compare our approach to other exertion measurement methods. Finally, we demonstrate and discuss its potential for automated, unobtrusive measurements and real-time game adaptation

Real-World Inter-Observer Variability of the Sequential Organ Failure Assessment (SOFA) Score in Intensive Care Medicine: the Time Has come for an Update: Authors’ Reply

info:eu-repo/semantics/publishedVersio

Current Methods for the Treatment and Prevention of Drug-Induced Parkinsonism and Tardive Dyskinesia in the Elderly

Drug-induced parkinsonism (DIP) and tardive dyskinesia (TD) are iatrogenic consequences of antidopaminergic drugs. Both are particularly prevalent among the elderly and those with dementia. However, despite their prevalence, these disorders are often overlooked. Both entities share risk factors, physiopathological mechanisms and, to some degree, therapeutic approaches. Withdrawing the causal agent, reducing the dose or switching to a less potent antidopaminergic drug should be the first therapeutic options. Here we review both entities and emerging therapies including the recently approved drugs deutetrabenazine and valbenazine. We discuss relevant aspects for clinical practice such as new diagnostic techniques and the latest advances in the understanding of DIP and TD

Voluntary control of wearable robotic exoskeletons by patients with paresis via neuromechanical modeling.

BACKGROUND: Research efforts in neurorehabilitation technologies have been directed towards creating robotic exoskeletons to restore motor function in impaired individuals. However, despite advances in mechatronics and bioelectrical signal processing, current robotic exoskeletons have had only modest clinical impact. A major limitation is the inability to enable exoskeleton voluntary control in neurologically impaired individuals. This hinders the possibility of optimally inducing the activity-driven neuroplastic changes that are required for recovery. METHODS: We have developed a patient-specific computational model of the human musculoskeletal system controlled via neural surrogates, i.e., electromyography-derived neural activations to muscles. The electromyography-driven musculoskeletal model was synthesized into a human-machine interface (HMI) that enabled poststroke and incomplete spinal cord injury patients to voluntarily control multiple joints in a multifunctional robotic exoskeleton in real time. RESULTS: We demonstrated patients' control accuracy across a wide range of lower-extremity motor tasks. Remarkably, an increased level of exoskeleton assistance always resulted in a reduction in both amplitude and variability in muscle activations as well as in the mechanical moments required to perform a motor task. Since small discrepancies in onset time between human limb movement and that of the parallel exoskeleton would potentially increase human neuromuscular effort, these results demonstrate that the developed HMI precisely synchronizes the device actuation with residual voluntary muscle contraction capacity in neurologically impaired patients. CONCLUSIONS: Continuous voluntary control of robotic exoskeletons (i.e. event-free and task-independent) has never been demonstrated before in populations with paretic and spastic-like muscle activity, such as those investigated in this study. Our proposed methodology may open new avenues for harnessing residual neuromuscular function in neurologically impaired individuals via symbiotic wearable robots

A comparison of methods for the quality control of 123I-Ioflupane

Objetivos: Se pretende establecer un método sencillo, preciso y reproducible para determinar la pureza radioquímica del 123I-Ioflupano.Material y Métodos: Se utilizó la radiocromatografía con tres tipos de fases estacionarias (whatmann-17MM, Whatmann-3MM e ITLC-SG) y cinco fases móviles (Éter Dietílico, Metiletilcetona, NaCl al 0,9%, Metanol:Cloroformo(1:9), Metanol:Agua al 85%). Este procedimiento fue realizado con alícuotas de 123I-Ioflupano (DatScan®) y 123INa para estudiar el comportamiento del yoduro libre.Resultados: Se descartó el whatmann-3MM por su mala separación y el ITLC-SG con Metanol:Agua por su baja reproducibilidad. Usando Metiletilcetona como fase móvil en ITLC-SG y Whatmann-17MM no se observa la fracción libre. Únicamente se observó la fracción libre en Whatmann-17MM tanto con Metanol:Cloroformo como con Éter Dietílico y en ITLC-SG con NaCl 0,9% .Conclusiones: La mejor separación del 123I-Ioflupano del 123I- fue observada con: Whatmann-17MM/ Metanol:Cloroformo (1:9) (Rf de la fracción ligada = 0,62±0,03, Pureza Radioquímica = 91,6±2,36) , ITLC-SG/NaCl 0,9% (Rf de la fracción ligada = 0,21±0,05, Pureza Radioquímica = 90,19±3,4) y Whatmann-17MM /Éter Dietílico (Rf de la fracción ligada = 0,62±0,06, Pureza Radioquímica = 92,79±1,65).Aim: The aim of this study is to stablish an easy, reproducible, and precise method to determine the radiochemical purity of 123I-Ioflupane.Material and methods: Different systems of radiochromatography were used with three types of stationary phases (Whatmann-17MM, Whatmann-3MM and ITLC-SG) and five mobile phases (Diethylether, NaCl 0.9%, Methanol:Chloroform(1:9), Methanol:Water (85:15) and Methylethylketone). This procedure was made with samples of 123I-Ioflupane and 123I-NaI to study the behaviour of free iodide.Results: Whatmann-3MM was rejected due to an unsuccessful separation and Methanol:Water (85:15) with ITLC-SG was not reproducible. Free fraction wasn`t observed in Methylethylketone as mobile phase with ITLC-SG and Whatmann-17MM. Free fraction was just observed in Whatmann-17MM with Methanol:Cloroform and Diethylether and in ITLC-SG with NaCl 0.9%.Conclusions: Maximum separation between 123I-Ioflupane and 123I- was observed in Whatmann-17MM/Methanol:Cloroform(1:9) (Rf of bound fraction = 0.62±0.03, Radiochemical Purity =91.6±2.36%), ITLC-SG/NaCl 0,9% (Rf of bound fraction = 0.21±0.05, Radiochemical Purity =90.19±3.4%) and Whatmann-17MM/ Diethylether (Rf of bound fraction = 0.62±0.06, Radiochemical Purity =92.79±1.65%)

Molecular Characterization of Growth Hormone-producing Tumors in the GC Rat Model of Acromegaly

D.A.C. was supported by the Nicolás Monardes program of the Andalusian Ministry of Health (C-0015-2014) and by a grant from the Andalusian Ministry of Science and Innovation (CTS-7478). A.S-M and A.L.C were supported by grants from the ISCIII-Subdirección General de Evaluación y Fomento de la Investigación co-funded with Fondos FEDER (PI12/0143 and PI13/02043, respectively) and the Andalusian Regional Government (CTS-444) and a grant from Pfizer Spain. R.L.C. was supported by a grant from Andalusian Ministry of Health (PI0302-2012). R.M.L. was supported by grants from Proyecto de Investigación en Salud (FIS) PI13- 00651 (funded by Instituto de Salud Carlos III), CTS-1406, PI-0639-2012, BIO-0139 (funded by Junta de Andalucía) and by Ayuda Merck Serono 2013. J. P. C. was funded by a grant (BFU2013-43282-R) from Ministerio de Economía y Competitividad. CIBER is an initiative of Instituto de Salud Carlos III, Ministerio de Sanidad, Servicios Sociales e Igualdad, Spain. J.F.M.R. is supported by the “Sara Borrell” program from the Instituto de Salud Carlos III. R.M. Luque and J.P. Castaño have received grants and lecture fees from Ipsen and Novartis. E. Venegas-Moreno and A. Soto-Moreno received grants and lecture fees from Ipsen, Novartis and Pfizer. A. Leal-Cerro received grants from Novartis and Pfizer. David Cano received a grant from Novartis

Prevalence of Cryptosporidium oocysts and Giardia cysts in raw and treated sewage sludges

Treated sludge from wastewater treatment plants (WWTPs) is commonly used in agriculture as fertilizers and to amend soils. The most significant health hazard for sewage sludge relates to the wide range of pathogenic microorganisms such as protozoa parasites.The objective of this study was to collect quantitative data on Cryptosporidium oocysts and Giardia cysts in the treated sludge in wastewater treatment facilities in Spain. Sludge from five WWTPs with different stabilization processes has been analysed for the presence of Cryptosporidium and Giardia in the raw sludge and after the sludge treatment. A composting plant (CP) has also been assessed. After a sedimentation step, sludge samples were processed and (oo)cysts were isolated by immunomagnetic separation (IMS) and detected by immunofluorescence assay (IFA). Results obtained in this study showed that Cryptosporidium oocysts and Giardia cysts were present in 26 of the 30 samples (86.6%) of raw sludge samples. In treated sludge samples, (oo)cysts have been observed in all WWTP's analysed (25 samples) with different stabilization treatment (83.3%). Only in samples from the CP no (oo)cysts were detected. This study provides evidence that (oo)cysts are present in sewage sludge-end products from wastewater treatment processes with the negative consequences for public health.We appreciate the financial support provided by Entidad Publica Saneamiento Aguas (EPSAR).Amoros Muñoz, I.; Moreno Trigos, MY.; Reyes-Sosa, MB.; Moreno-Mesonero, L.; Alonso Molina, JL. (2016). Prevalence of Cryptosporidium oocysts and Giardia cysts in raw and treated sewage sludges. Environmental Technology. 37(22):2898-2904. doi:10.1080/09593330.2016.1168486S28982904372