The importance of pH in regulating the function of the Fasciola hepatica cathepsin L1 cysteine protease

The helminth parasite Fasciola hepatica secretes cathepsin L cysteine proteases to invade its host, migrate through tissues and digest haemoglobin, its main source of amino acids. Here we investigated the importance of pH in regulating the activity and functions of the major cathepsin L protease FheCL1. The slightly acidic pH of the parasite gut facilitates the auto-catalytic activation of FheCL1 from its inactive proFheCL1 zymogen; this process was approximately 40-fold faster at pH 4.5 than at pH 7.0. Active mature FheCL1 is very stable at acidic and neutral conditions (the enzyme retained approximately 45% activity when incubated at 37 degrees C and pH 4.5 for 10 days) and displayed a broad pH range for activity peptide substrates and the protein ovalbumin, peaking between pH 5.5 and pH 7.0. This pH profile likely reflects the need for FheCL1 to function both in the parasite gut and in the host tissues. FheCL1, however, could not cleave its natural substrate Hb in the pH range pH 5.5 and pH 7.0; digestion occurred only at pH

Micro-environmental conditions modulate protein secretion and infectivity of the Trichinella spiralis L1 larva

After digestion of infected meat the free L1 of Trichinella spp. penetrate the intestinal mucosa where they moult to the mature adult stage. We have used proteomics to identify changes in protein secretion during in vitro culture of free T. spiralis muscle larvae under different environmental conditions, and to correlate these changes with their infectivity in mice. Muscle larvae were cultured in different media (RPMI-1640, C-199 and HBSS) under conditions of anaerobiosis, microaerobiosis and in 5% CO(2) at 37 degrees C. Following incubation the larval excretory/secretory proteins were analysed by two-dimensional gel electrophoresis and the larvae were used to orally infect naïve CD1 mice. For all culture media tested, infectivity of the L1 was preserved following incubation in anaerobic conditions. In contrast, the infectivity of worms cultured in nutrient-rich media was almost completely abolished in both microaerobiosis and in the presence of 5% CO(2). Some infectivity was retained in poor or reduced culture media. Comparative analysis of larval infectivity and protein secretion showed that loss of infectivity correlated with the appearance of non-tyvelosylated proteins that in turn may be related to the onset of moulting.

Proteomic analysis of the excretory-secretory proteins of the Trichinella spiralis L1 larva, a nematode parasite of skeletal muscle

Trichinella spiralis is an intracellular nematode parasite of mammalian skeletal muscle. Infection of the muscle cell leads to the formation of a host-parasite complex that results in profound alterations to the host cell and a re-alignment of muscle-specific gene expression. The role of parasite excretory-secretory (ES) proteins in mediating these effects is currently unknown, largely due to the difficulty in identifying and assigning function to individual proteins. In this study, a global proteomics approach was used to analyse the ES proteins from T. spiralis muscle larvae. Following 2-DE of ES proteins,MALDI-TOF-MS and LC-MS/MS were used to identify the peptide spots. Specific Trichinella EST databases were assembled and used to analyse the data. Despite the current absence of a Trichinella genome-sequencing project, 43 out of 52 protein spots analysed were identified and included the major secreted glycoproteins. Other novel proteins were identified from matches with sequences in the T. spiralis database. Our results demonstrate the value of proteomics as a tool for the identification of Trichinella ES proteins and in the study of the molecular mechanism underpinning the formation of the host-parasite complex during Trichinella infections.

A multi-output two-stage locally regularized model construction method using the extreme learning machine

This paper investigates the construction of linear-in-the-parameters (LITP) models for multi-output regression problems. Most existing stepwise forward algorithms choose the regressor terms one by one, each time maximizing the model error reduction ratio. The drawback is that such procedures cannot guarantee a sparse model, especially under highly noisy learning conditions. The main objective of this paper is to improve the sparsity and generalization capability of a model for multi-output regression problems, while reducing the computational complexity. This is achieved by proposing a novel multi-output two-stage locally regularized model construction (MTLRMC) method using the extreme learning machine (ELM). In this new algorithm, the nonlinear parameters in each term, such as the width of the Gaussian function and the power of a polynomial term, are firstly determined by the ELM. An initial multi-output LITP model is then generated according to the termination criteria in the first stage. The significance of each selected regressor is checked and the insignificant ones are replaced at the second stage. The proposed method can produce an optimized compact model by using the regularized parameters. Further, to reduce the computational complexity, a proper regression context is used to allow fast implementation of the proposed method. Simulation results confirm the effectiveness of the proposed technique. © 2013 Elsevier B.V.

The prognostic significance of PD-L1 in bladder cancer

Immunotherapy is a promising strategy for the treatment of various types of cancer. An antibody that targets programmed death ligand-1 (PD-L1) pathway has been shown to be active towards various types of cancer, including melanoma and lung cancer. MPDL3280A, an anti‑PD-L1 antibody, has shown clear clinical activity in PD-L1-overexpressing bladder cancer with an objective response rate of 40-50%, resulting in a breakthrough therapy designation granted by FDA. These events pronounce the importance of targeting the PD-L1 pathway in the treatment of bladder cancer. In the present study, we investigated the prognostic significance of the expression of three genes in the PD-L1 pathway, including PD-L1, B7.1 and PD-1, in three independent bladder cancer datasets in the Gene Expression Omnibus database. PD-L1, B7.1 and PD-1 were significantly associated with clinicopathological parameters indicative of a more aggressive phenotype of bladder cancer, such as a more advanced stage and a higher tumor grade. In addition, a high level expression of PD-L1 was associated with reduced patient survival. Of note, the combination of PD-L1 and B7.1 expression, but not other combinations of the three genes, were also able to predict patient survival. Our findings support the development of anti-PD-L1, which blocks PD-L1-PD-1 and B7.1-PD-L1 interactions, in treatment of bladder cancer. The observations were consistent in the three independent bladder cancer datasets consisting of a total of 695 human bladder specimens. The datasets were then assessed and it was found that the expression levels of the chemokine CC-motif ligand (CCL), CCL3, CCL8 and CCL18, were correlated with the PD-L1 expression level, while ADAMTS13 was differentially expressed in patients with a different survival status (alive or deceased). Additional investigations are required to elucidate the role of these genes in the PD-L1-mediated immune system suppression and bladder cancer progression. In conclusion, findings of this study suggested that PD-L1 is an important prognostic marker and a therapeutic target for bladder cancer.

Immune-derived PD-L1 gene expression defines a subgroup of stage II/III colorectal cancer patients with favorable prognosis that may be harmed by adjuvant chemotherapy

A recent phase 2 study of metastatic colorectal carcinoma (CRC) patients showed that mismatch repair gene status was predictive of clinical response to PD-1-targeting immune checkpoint blockade. Further examination revealed strong correlation between PD-L1 protein expression and microsatellite instability (MSI) in stage IV CRC, suggesting that the amount of PD-L1 protein expression could identify late stage patients who may benefit from immunotherapy. To assess whether the clinical associations between PD-L1 gene expression and MSI identified in metastatic CRC are also present in stage II/III CRC, we used in silico analysis to elucidate the cell types expressing the PD-L1 gene. We found a significant association of PD-L1 gene expression with MSI in early stage CRC (P < 0.001) and show that unlike in non-CRC tumors, PD-L1 is derived predominantly from the immune infiltrate. We demonstrate that PD-L1 gene expression has positive prognostic value in the adjuvant disease setting (PD-L1low v PD-L1high HR = 9.09; CI, 2.11-39.10). PD-L1 gene expression had predictive value, as patients with high PD-L1 expression appear to be harmed by standard-of-care treatment (HR = 4.95; CI,1.10-22.35). Building on the promising results from the metastatic CRC PD-1-targeting trial, we provide compelling evidence that PD-L1high/MSI/immunehigh stage II/III CRC patients should not receive standard chemotherapy. This conclusion supports the rationale to clinically evaluate this patient subgroup for PD-1 blockade treatment.

LSP communication in a digital age: the outcomes of teaching and learning in tertiary education

This paper discusses the changes brought by the communication revolution in teaching and learning in the scope of LSP. Its aim is to provide an insight on how teaching which was bi-dimensional, turned into a multidimensional system, gathering other complementary resources that have transformed, in a incredibly short time, the ways we receive share and store information, for instance as professionals, and keep in touch with our peers. The increasing rise of electronic publications, the incredible boom of social and professional networks, search engines, blogs, list servs, forums, e-mail blasts, Facebook pages, YouTube contents, Tweets and Apps, have twisted the way information is conveyed. Classes ceased to be predictable and have been empowered by digital platforms, innumerous and different data repositories (TILDE, IATE, LINGUEE, and so many other terminological data banks) that have definitely transformed the academic world in general and tertiary education in particular. There is a bulk of information to be digested by students, who are no longer passive but instead responsible and active for their academic outcomes. The question is whether they possess the tools to select only what is accurate and important for a certain subject or assignment, due to that overflow? Due to the reduction of the number of course years in most degrees, after the implementation of Bologna and the shrinking of the curricula contents, have students the possibility of developing critical thinking? Both teaching and learning rely on digital resources to improve the speed of the spreading of knowledge. But have those changes been effective to promote really communication? Furthermore, with the increasing Apps that have already been developed and will continue to appear for learning foreign languages, for translation among others, will the students feel the need of learning them once they have those Apps. These are some the questions we would like to discuss in our paper.

La eficacidad de los cursos de ELE : la percepción de los usuarios de L1/L2

Cette recherche vise à étudier l'efficacité des cours d'espagnol comme langue étrangère, à partir de la perception des étudiants d’espagnol comme langue d’origine. Pour atteindre les résultats et les conclusions obtenus, cette étude qualitative compte sur la participation des répondants hispaniques qui ont appris l'espagnol à la maison, à l'école primaire, au secondaire, au cégep et/ou au collège. Les participants ont répondu à un questionnaire qui leur a permis d'exprimer leur perception à l'égard des cours d’espagnol comme langue étrangère qu'ils ont pris au cours de leur étude primaire, secondaire ou supérieure. Les résultats de cette recherche indiquent que les élèves hispaniques ne sont pas intégrés dans les classes ELE, ce qui motive les participants à poursuivre l’étude de la langue espagnole. Comme expliqué par Gardner (1985) dans sa théorie du modèle socio-éducationnel, l’acquisition des langages intervient dans un contexte culturel spécifique et elle est influencée par la croyance populaire et les différences individuelles. Ces dernières sont intégrées dans le concept du motif d'intégration, qui est composé du sens de l'intégration, de l'attitude envers la situation d'apprentissage et de la motivation. Les réponses données par les participants dans cette étude peuvent être utiles à la conception et au développement de cours et de méthodes d’apprentissage de l’espagnol pour les hispanophones d’origine. En tenant compte de leurs points de vue, les étudiantes auraient un sens plus élevé de l’intégration, ce qui influence directement leur motivation et par le fait même leur processus d’apprentissage.

Die Rolle des Zelladhäsionsmoleküls L1 in der Tumorprogression

Das neuronale Adhäsionsmolekül L1 wird neben den Zellen des Nervensystems auf vielen humanen Tumoren exprimiert und ist dort mit einer schlechten Prognose für die betroffenen Patienten assoziiert. Zusätzlich zu seiner Funktion als Oberflächenmolekül kann L1 durch membranproximale Spaltung in eine lösliche Form überführt werden. In der vorliegenden Arbeit wurde der Einfluss von L1 auf die Motilität von Tumorzellen untersucht. Lösliches L1 aus Asziten führte zu einer Integrin-vermittelten Zellmigration auf EZM-Substraten. Derselbe Effekt wurde durch Überexpression von L1 in Tumorlinien beobachtet. Weiterhin führt die L1-Expression zu einer erhöhten Invasion, einem verstärkten Tumorwachstum in NOD/SCID Mäusen und zur konstitutiven Aktivierung der MAPK ERK1/2. Eine Mutation in der zytoplasmatischen Domäne von hL1 (Thr1247Ala/Ser1248Ala)(hL1mut) führte hingegen zu einer Blockade dieser Funktionen. Dies weist daraufhin, dass nicht nur lösliches L1, sondern auch die zytoplasmatische Domäne von L1 funktionell aktiv ist. Im zweiten Teil der Arbeit wurde der Mechanismus, der L1-vermittelten Signaltransduktion untersucht. Die zytoplasmatische Domäne von L1 gelangt nach sequenzieller Proteolyse durch ADAM und Presenilin-abhängiger γ-Sekretase Spaltung in den Zellkern. Diese Translokation im Zusammenspiel mit der Aktivierung der MAPK ERK1/2 durch L1-Expression führt zu einer L1-abhängigen Genregulation. Die zytoplasmatische Domäne von hL1mut konnte ebenfalls im Zellkern detektiert werden, vermittelte jedoch keine Genregulation und unterdrückte die ERK1/2 Phosphorylierung. Die L1-abhängige Induktion von ERK1/2-abhängigen Genen wie Cathepsin B, β3 Integrin und IER 3 war in Zellen der L1-Mutante unterdrückt. Die Expression des Retinsäure-bindenden Proteins CRABP-II, welches in hL1 Zellen supprimiert wird, wurde in der L1-Mutante nicht verändert. Weitere biochemische Untersuchungen zeigen, dass die zytoplasmatische Domäne von L1 Komplexe mit Transkriptionsfaktoren bilden kann, die an Promoterregionen binden können. Die dargestellten Ergebnisse belegen, dass L1-Expression in Tumoren an drei Funktionen beteiligt ist; (i) L1 erhöht Zellmotilität, (ii) fördert Tumorprogression durch Hochregulation von pro-invasiven und proliferationsfördernden Genen nach Translokation in den Nukleus und (iii) schützt die Zellen mittels Regulation pro- bzw. anti-apoptotischer Gene vor Apoptose. Die mutierte Phosphorylierungsstelle im L1-Molekül ist essentiell für diese Prozesse. Die Anwendung neuer Therapien für Patienten mit L1-positiven Karzinomen kann mit Hinblick auf die guten Erfolge der Antikörper-basierenden Therapie mit dem mAk L1-11A diskutiert werden.

The improvement of reading skills of L1 and ESL children using a Response to Intervention (RtI) Model

Resumen tomado de la publicaci??n

Functional studies of the deubiquitinating enzymes USP19, USP4 and UCH-L1

El marcaje de proteínas con ubiquitina, conocido como ubiquitinación, cumple diferentes funciones que incluyen la regulación de varios procesos celulares, tales como: la degradación de proteínas por medio del proteosoma, la reparación del ADN, la señalización mediada por receptores de membrana, y la endocitosis, entre otras (1). Las moléculas de ubiquitina pueden ser removidas de sus sustratos gracias a la acción de un gran grupo de proteasas, llamadas enzimas deubiquitinizantes (DUBs) (2). Las DUBs son esenciales para la manutención de la homeostasis de la ubiquitina y para la regulación del estado de ubiquitinación de diferentes sustratos. El gran número y la diversidad de DUBs descritas refleja tanto su especificidad como su utilización para regular un amplio espectro de sustratos y vías celulares. Aunque muchas DUBs han sido estudiadas a profundidad, actualmente se desconocen los sustratos y las funciones biológicas de la mayoría de ellas. En este trabajo se investigaron las funciones de las DUBs: USP19, USP4 y UCH-L1. Utilizando varias técnicas de biología molecular y celular se encontró que: i) USP19 es regulada por las ubiquitin ligasas SIAH1 y SIAH2 ii) USP19 es importante para regular HIF-1α, un factor de transcripción clave en la respuesta celular a hipoxia, iii) USP4 interactúa con el proteosoma, iv) La quimera mCherry-UCH-L1 reproduce parcialmente los fenotipos que nuestro grupo ha descrito previamente al usar otros constructos de la misma enzima, y v) UCH-L1 promueve la internalización de la bacteria Yersinia pseudotuberculosis.

Consideraciones pragmáticas sobre la cortesía y su tratamiento en la enseñanza del español L1.

Resumen en inglés

Enseñar nuevas lenguas en la escuela: L1, L2, LE..., NL.

Resumen basado en el de la publicación. Número monográfico con el título: La enseñanza aprendizaje del español como segunda lengua (L2) en contextos educativos multilingües

Cognitive processes in L1 and L2 writing : a cross-sectional study. 'Procesos cognitivos en la escritura de L1 y L2 : un estudio transversal'.

Conocer en qué medida el grado de dominio de la L2 configura el tiempo asignado a los procesos de escritura y su distribución en el transcurso de la composición. 21 aprendices de inglés como lengua extranjera divididos en tres niveles de dominio: pre-intermedio (N1), intermedio (N2) y avanzado (N3), estando integrado cada grupo por 7 sujetos de tercero de BUP, tercero de Magisterio de Lenguas Extranjeras y recién licenciados en Filología Inglesa respectivamente. Cada sujeto de la muestra realizó una composición argumentada en L1 y L2 siguiendo la técnica del pensamiento en voz alta en dos sesiones distintas y recibiendo entrenamiento previo en la técnica de pensar en voz alta. La tarea en L2 se llevó a cabo antes de la de L1 para tratar de evitar la traducción dada la similitud de los temas propuestos, idénticos para todos los sujetos. Al finalizar los sujetos rellenaron un cuestionario restrospectivo. Se grabaron y transcribieron las sesiones. Oxford Placement Test, prueba que se centra en la discriminación de habilidades gramaticales de lectura y de comprensión auditiva. Análisis de la distribución temporal de los procesos de planificación y formulación en composiciones argumentales en L1 y L2. ANOVA. Los sujetos de la muestra dedicaron un porcentaje similar de tiempo a formular globalmente sus textos en L1 y L2, sin embargo, la proporción de procesos de formulación fluída frente a los de naturaleza problemática varían según la lengua: en L1 la proporción en de 5/1 y en L2 de 2/1. A medida que aumenta la habilidad del escritor se observa una tendencia a asumir progresivamente el papel de escritor como controlador del proceso, en detrimento del escritor como productor de un texto. Los resultados de la investigación apuntan a que no puede considerarse que el proceso de composición sea lineal, aunque sea incorrecto pensar que el escritor va a dedicar idénticos recursos temporales al mismo proceso en todas las partes de la composición (planificación, formulación y revisión). Existe un punto intermedio entre los dos extremos: ciertos procesos tienen más probabilidad de ser activados en ciertos momentos que otros y, como consecuencia, su probabilidad de ocurrencia cambia a medida que se avanza en el transcurso de la tarea.